{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001094","ANALYSIS_ID":"AN001780","VERSION":"1","CREATED_ON":"November 13, 2018, 6:58 pm"},

"PROJECT":{"PROJECT_TITLE":"Adverse effects of PAHs on lung cells","PROJECT_SUMMARY":"Low-molecular-weight (LMW) polycyclic aromatic hydrocarbons (PAHs) are more prevalent in the environment, occupational settings, as well as in secondhand smoke (SHS), when compared to their high molecular weight counterparts, such as benzo[a]pyrene (B[a]P). Previously, we demonstrated that SHS-prevalent LMW PAHs activate p38-MAPK-dependent dysregulation of gap junction intercellular communication (GJIC) and increased cytokines involved in inflammatory lung diseases. However, there is little known about the early mechanistic events leading to inflammation, specifically those mediated through lipid signaling and eicosanoids. Secondhand smoke is a complex mixture and to model this feature in vitro we examined the effects of a binary mixture of 1-methylanthracene (1-MeA) and fluoranthene (Flthn) in C10 cells, a mouse, non-tumorigenic alveolar type II cell line via a global metabolomics approach to evaluate the lipids.","INSTITUTE":"University of Colorado, Denver","DEPARTMENT":"Pharmaceutical Sciences, Anschutz Medical Campus","LAST_NAME":"Bauer","FIRST_NAME":"Alison","ADDRESS":"12850 E. Montview Dr. Rm 3125, Aurora, CO 80045","EMAIL":"alison.bauer@ucdenver.edu","PHONE":"303-724-6297","FUNDING_SOURCE":"R15 ES 024893-01 (AKB) and the Flight Attendant Medical Research Institute (FAMRI) CIA 130022 (AKB).","PROJECT_COMMENTS":"There are two studies that will be uploaded. AKB study 1 and AKB study 2.","CONTRIBUTORS":"Katelyn J. Siegrist, DeeDee Romo, Brad L. Upham, Michael Armstrong, Kevin Quinn, Lauren Vanderlinden, Kalpana Velmurugan, Mark Elie, Dominik Reinhold, Nichole Reisdorph, Laura Saba"},

"STUDY":{"STUDY_TITLE":"Early Mechanistic Events Induced by Secondhand Smoke Prevalent Low Molecular Weight Polycyclic Aromatic Hydrocarbons in Mouse Lung Epithelial Cells (part-I)","STUDY_SUMMARY":"We evaluated lung epithelial cells exposed to low molecular weight polycyclic aromatic hydrocarbons and what lipid metabolites were produced following early exposure, prior to metabolism. We used 40 uM 1-methylanthracene and fluoranthene (1:1 ratio)for 30 min, 1h, and 4 h time points for the global untargeted metabolomics study (AKB study 1).","INSTITUTE":"University of Colorado, Denver","DEPARTMENT":"Anschutz Medical Campus","LAST_NAME":"Bauer","FIRST_NAME":"Alison","ADDRESS":"12850 E. Montview Dr. Rm 3125, Aurora, CO 80045","EMAIL":"alison.bauer@ucdenver.edu","PHONE":"303-724-6297"},

"SUBJECT":{"SUBJECT_TYPE":"Cultured cells","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"sample.1",
"Factors":{"Treatment":"DMSO","Time":"30 min"}
},
{
"Subject ID":"-",
"Sample ID":"sample.2",
"Factors":{"Treatment":"DMSO","Time":"30 min"}
},
{
"Subject ID":"-",
"Sample ID":"sample.3",
"Factors":{"Treatment":"DMSO","Time":"30 min"}
},
{
"Subject ID":"-",
"Sample ID":"sample.4",
"Factors":{"Treatment":"DMSO","Time":"30 min"}
},
{
"Subject ID":"-",
"Sample ID":"sample.5",
"Factors":{"Treatment":"2PAHs","Time":"30 min"}
},
{
"Subject ID":"-",
"Sample ID":"sample.6",
"Factors":{"Treatment":"2PAHs","Time":"30 min"}
},
{
"Subject ID":"-",
"Sample ID":"sample.7",
"Factors":{"Treatment":"2PAHs","Time":"30 min"}
},
{
"Subject ID":"-",
"Sample ID":"sample.8",
"Factors":{"Treatment":"2PAHs","Time":"30 min"}
},
{
"Subject ID":"-",
"Sample ID":"sample.9",
"Factors":{"Treatment":"DMSO","Time":"1 hr"}
},
{
"Subject ID":"-",
"Sample ID":"sample.10",
"Factors":{"Treatment":"DMSO","Time":"1 hr"}
},
{
"Subject ID":"-",
"Sample ID":"sample.11",
"Factors":{"Treatment":"DMSO","Time":"1 hr"}
},
{
"Subject ID":"-",
"Sample ID":"sample.12",
"Factors":{"Treatment":"DMSO","Time":"1 hr"}
},
{
"Subject ID":"-",
"Sample ID":"sample.13",
"Factors":{"Treatment":"2PAHs","Time":"1 hr"}
},
{
"Subject ID":"-",
"Sample ID":"sample.14",
"Factors":{"Treatment":"2PAHs","Time":"1 hr"}
},
{
"Subject ID":"-",
"Sample ID":"sample.15",
"Factors":{"Treatment":"2PAHs","Time":"1 hr"}
},
{
"Subject ID":"-",
"Sample ID":"sample.16",
"Factors":{"Treatment":"2PAHs","Time":"1 hr"}
},
{
"Subject ID":"-",
"Sample ID":"sample.17",
"Factors":{"Treatment":"DMSO","Time":"4 hr"}
},
{
"Subject ID":"-",
"Sample ID":"sample.18",
"Factors":{"Treatment":"DMSO","Time":"4 hr"}
},
{
"Subject ID":"-",
"Sample ID":"sample.19",
"Factors":{"Treatment":"DMSO","Time":"4 hr"}
},
{
"Subject ID":"-",
"Sample ID":"sample.20",
"Factors":{"Treatment":"DMSO","Time":"4 hr"}
},
{
"Subject ID":"-",
"Sample ID":"sample.21",
"Factors":{"Treatment":"2PAHs","Time":"4 hr"}
},
{
"Subject ID":"-",
"Sample ID":"sample.22",
"Factors":{"Treatment":"2PAHs","Time":"4 hr"}
},
{
"Subject ID":"-",
"Sample ID":"sample.23",
"Factors":{"Treatment":"2PAHs","Time":"4 hr"}
},
{
"Subject ID":"-",
"Sample ID":"sample.24",
"Factors":{"Treatment":"2PAHs","Time":"4 hr"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"For the global metabolomics with a lipid focus, immediately following treatment, cell monolayers were washed 3 times with cold PBS, and cells were scraped and fixed in 500 μL 70% MeOH. Samples were stored immediately at -80°C until pretreatment for solid phase extraction (SPE).","COLLECTION_PROTOCOL_FILENAME":"AKB_Untargeted-protocol.docx","COLLECTION_PROTOCOL_COMMENTS":"For AKB study 1 protocol, the collection, treatment, and sample prep are all in the same file.","SAMPLE_TYPE":"Epithelial cells"},

"TREATMENT":{"TREATMENT_SUMMARY":"For the global metabolomics with a focus on the lipids: Cells (passage <20) were maintained in CMRL 1066 media containing 10% FBS and 1% glutamate in a humidified atmosphere at 37˚ C, 5% CO2, and 95% air (Osgood et al. 2013). Once cells were grown to confluence, cells were serum deprived for 24 hr prior to treatment with the binary 1:1 LMW PAH mixture of 1-MeA and Flthn at a final dose of 40 μM. This mixture has been used previously in our lab (PMC5808746) to represent two LMW PAH’s common in secondhand smoke and environmental exposures.","TREATMENT_PROTOCOL_FILENAME":"AKB_Untargeted-protocol","TREATMENT_PROTOCOL_COMMENTS":"For AKB study 1 protocol, the collection, treatment, and sample prep are all in the same file."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"The hydrophobic fraction was extracted from the samples using an organic liquid-liquid extraction technique with methyl tert-butyl ether (MTBE) and water (Cruickshank-Quinn et al. 2014; Yang et al. 2013). The upper hydrophobic layer was transferred to a new tube. The hydrophobic layer was then dried under nitrogen, reconstituted in 200 µL of methanol, and stored at -80 °C until LCMS analysis.","SAMPLEPREP_PROTOCOL_FILENAME":"AKB_Targeted-protocol.docx","SAMPLEPREP_PROTOCOL_COMMENTS":"For AKB study 1 protocol, the collection, treatment, and sample prep are all in the same file."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Agilent 1290","COLUMN_NAME":"Agilent Zorbax Rapid Resolution HD SB-C18"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"MS_COMMENTS":"-","INSTRUMENT_NAME":"Agilent 6210 TOF","INSTRUMENT_TYPE":"TOF","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_RESULTS_FILE":"ST001094_AN001780_Results.txt UNITS:abundance/counts Has m/z:No Has RT:Yes RT units:Minutes"}

}