{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST000656","ANALYSIS_ID":"AN001001","VERSION":"1","CREATED_ON":"June 26, 2017, 10:32 am"},

"PROJECT":{"PROJECT_TITLE":"Omega-6 and omega-3 oxylipins are implicated in soybean oil-induced obesity in mice","PROJECT_SUMMARY":"In this study we have compared the metabolic effects of conventional soybean oil to those of genetically modified Plenish soybean oil, that is low in linoleic acid and high in oleic acid. This work builds on our previous study showing that soybean oil, rich in polyunsaturated fats, is more obesogenic and diabetogenic than coconut oil, rich in saturated fats (PMID: 26200659). Here, in order to elucidate the mechanisms responsible for soybean oil induced obesity, we have performed the first ever metabolomics (in plasma and liver) and proteomics on the livers of mice fed the two soybean oil diets (plus those fed a high coconut oil and Viv chow diet). Our results show that the new high oleic soybean oil induces less obesity and adiposity than conventional soybean oil, but can cause hepatomegaly and liver dysfunction. Metabolomic analysis reveals that the hepatic and plasma metabolic profiles differ considerably between the two soybean oils. Hepatic C18 oxylipin metabolites of omega-6 (ω6) and omega-3 (ω3) fatty acids (linoleic and α-linolenic acid, respectively) in the cytochrome P450/soluble epoxide hydrolase pathway were found to correlate positively with obesity.","INSTITUTE":"University of California, Riverside","DEPARTMENT":"Cell Biology and Neuroscience","LAST_NAME":"Sladek","FIRST_NAME":"Frances","ADDRESS":"2115 Biological Sciences Building,University of California, Riverside, CA 92521-0314","EMAIL":"frances.sladek@ucr.edu","PHONE":"951-827-2264"},

"STUDY":{"STUDY_TITLE":"Omega-6 and omega-3 oxylipins are implicated in soybean oil-induced obesity in mice (part III)","STUDY_SUMMARY":"In this study we have compared the metabolic effects of conventional soybean oil to those of genetically modified Plenish soybean oil, that is low in linoleic acid and high in oleic acid. This work builds on our previous study showing that soybean oil, rich in polyunsaturated fats, is more obesogenic and diabetogenic than coconut oil, rich in saturated fats (PMID: 26200659). Here, in order to elucidate the mechanisms responsible for soybean oil induced obesity, we have performed the first ever metabolomics (in plasma and liver) and proteomics on the livers of mice fed the two soybean oil diets (plus those fed a high coconut oil and Viv chow diet). Our results show that the new high oleic soybean oil induces less obesity and adiposity than conventional soybean oil, but can cause hepatomegaly and liver dysfunction. Metabolomic analysis reveals that the hepatic and plasma metabolic profiles differ considerably between the two soybean oils. Hepatic C18 oxylipin metabolites of omega-6 (ω6) and omega-3 (ω3) fatty acids (linoleic and α-linolenic acid, respectively) in the cytochrome P450/soluble epoxide hydrolase pathway were found to correlate positively with obesity.","INSTITUTE":"University of California, Davis","DEPARTMENT":"Genome and Biomedical Sciences Facility","LABORATORY":"WCMC Metabolomics Core","LAST_NAME":"Fiehn","FIRST_NAME":"Oliver","ADDRESS":"1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616","EMAIL":"ofiehn@ucdavis.edu","PHONE":"(530) 754-8258"},

"SUBJECT":{"SUBJECT_TYPE":"Animal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090","GENOTYPE_STRAIN":"C57/BL6N","GENDER":"Male","ANIMAL_HOUSING":"SPF facility","ANIMAL_LIGHT_CYCLE":"12:12 h light-dark cycle","ANIMAL_FEED":"Ad libitum","ANIMAL_WATER":"Ad libitum"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"CSH_BioRec_01_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"N/A","TIME_POINT":"N/A","Genotype":"N/A"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_BioRec_02_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"N/A","TIME_POINT":"N/A","Genotype":"N/A"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_BioRec_03_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"N/A","TIME_POINT":"N/A","Genotype":"N/A"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_PM D3-17 e1_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"LA-HFD","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_PM D3-17 e2_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"LA-HFD","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_PM D3-17 e3reinject_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"LA-HFD","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_PM D3-17 e4_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"LA-HFD","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_PM D3-18 e1_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"LA-HFD","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_PM D3-18 e2reinject_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"LA-HFD","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_PM D3-18 e3_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"LA-HFD","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_PM D3-18 e4_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"LA-HFD","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_PM D4-16 e1_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"HFD","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_PM D4-16 e2_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"HFD","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_PM D4-16 e3_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"HFD","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_PM D4-16 e4_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"HFD","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_PM D4-17 e1_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"HFD","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_PM D4-17 e2_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"HFD","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_PM D4-18 e1_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"HFD","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_PM D4-18 e4_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"HFD","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_PM PL-4 e1_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"PL-HFD","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_PM PL-4 e3_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"PL-HFD","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_PM PL-4 e4_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"PL-HFD","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_PM PL-5 e1_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"PL-HFD","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_PM PL-5 e2_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"PL-HFD","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_PM PL-5 e3_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"PL-HFD","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_PM PL-6 e1_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"PL-HFD","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_PM PL-6 e4_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"PL-HFD","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_Viv-1_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"Viv chow","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_Viv-2_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"Viv chow","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_Viv-4_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"Viv chow","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_Viv-5_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"Viv chow","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_Viv-6_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"Viv chow","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_Viv-7_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"Viv chow","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_Viv-8_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"Viv chow","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
},
{
"Subject ID":"-",
"Sample ID":"CSH_Viv-9_pos mode_MS_07212014.d",
"Factors":{"GROUP_DESCRIPTION":"Viv chow","TIME_POINT":"24 weeks","Genotype":"WT C57-Bl6"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Liver tissue from mice on the diets for 24 weeks for metabolomic analysis was collected, rinsed in cold PBS, excess fluid was blotted with a kim-wipe and tissue was immediately snap frozen in liquid nitrogen before storage at -80°C. Blood was collected by cardiac puncture and centrifuged at 9 rcf for 5 min at 4°C. Plasma was stored immediately at -20°C."},

"TREATMENT":{"TREATMENT_SUMMARY":"Male C57/BL6N mice weaned at 3weeks of age were randomly assigned to one of the four diets: 1) VIV chow- normal rodent chow , low in fat and high in fiber 2) HFD (referred to as CO in the manuscript) -40 kcal% high fat diet with 36 kcal% from coconut oil and 4 kcal% from conventional soybean oil 3) LA-HFD (referred to as SO+CO in the manuscript) - 40 kcal% high fat diet with 21 kcal% fat calories from coconut oil and 19 kcal% from conventional soybean oil, of which 10 kcal% were from LA 4) PL-HFD (referred to as PL+CO in the manuscript) -40kcal% high fat diet in which conventional soybean oil in LA-HFD was replaced on a per gram basis with the genetically modified (GM) High Oleic Soybean Oil , Plenish"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"1) Keep Specimen on Dry Ice 2) Transfer Tissue Contents into a new 1.5mL labeled eppendorf tube; keep on dry ice at all times 3) Add three (3) 3mm metal grinding balls to each sample; store in -80C for 10minutes 4) Homogenize the entire tissue to fine powder using genogrinder; make sure that the metal grinding balls are ice-cold prior to homogenization (step 3) 5) Upon completion of homogenization, keep samples on dry ice 6) Weight out two (2) aliquots: a ~5mg aliquot for CSH_lipidomics and a ~4mg aliquot for Primary Metabolites by GCTOF a. Record the exact weight weighed out for each sample b. Keep all samples on dry ice 7) KEEP remaining tissue specimen (>90mg) for analysis of Oxylipins (store in -80C) 8) Analysis of Primary Metabolites (GCTOFMS) a. Add 1mL of ice-cold “degassed” 3:3:2 ACN/IPA/H2O b. Vortex for 10seconds c. Shake on shaker for 20min at -4C d. Centrifuge the samples for 2min at 14,000 rcf e. Transfer two (2) 500μL aliquots to new 1.5mL eppendorf tubes; one for backup the other to be dried to dryness using the SpeedVac f. IMPORTANT: The precipitated protein will be used for analysis of the proteome, DO NOT DISCARD THESE; Place these in a separate labeled box and store in -20C g. Keep all samples on ice during extraction period h. Dry down one (1) 500μL aliquot to complete dryness i. Perform cleanup on dried aliquot using 500μL of 50/50 v/v ACN/H2O j. Transfer supernatant and dry to completeness k. Submit for Derivatization 9) Analysis of Complex Lipids (LCQTOF) a. Add 225μL of ice-cold “degassed” MeOH containing “ISTD mixture” to homogenized 5mg aliquot b. Vortex for 10 seconds c. Add 750μL of ice-cold “degassed” MTBE containing 22:1 CE ISTD d. Vortex for 10 seconds e. Shake on Orbital Mixer for 6min at 4C f. Add 188μL of room temperature H2O g. Vortex for 20 seconds h. Centrifuge for 2min at 14,000 rcf i. Transfer two (2) aliquots of 350μL of top layer, one for backup stored in -20C, the other for analysis j. Keep bottom layer and store in -20C k. Dry down one (1) 350μL aliquot to dryness using the Speedvac l. Resuspend samples in 108.6μL of 50ng/mL CUDA m. Vortex and sonicate for 5minutes n. Centrifuge for 2min at 14,000 rcf o. Transfer 90μL to an amber vial with micro-insert (non-diluted) p. Transfer 10μL to a new 1.5mL eppendorf tube, dilute 20X with 50ng/mL CUDA in 90:10 MeOH:Toluene (10μL + 190μL CUDA) and transfer 100μL to amber vial with micro-insert (diluted for TGs) i. The dilution is based off previous experiences with liver samples","SAMPLEPREP_PROTOCOL_FILENAME":"SP_Extraction_Protocol_for_liver_multi-omic.pdf"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Agilent 6530","COLUMN_NAME":"Waters Acquity CSH C18 (100 x 2.1mm, 1.7um) 1.7um Pre-Column","FLOW_GRADIENT":"15% B to 99% B","FLOW_RATE":"0.6 mL/min","COLUMN_TEMPERATURE":"65 C","METHODS_FILENAME":"Data_Dictionary_Fiehn_laboratory_GCTOF_MS_primary_metabolism_10-15-2013_general.pdf","SOLVENT_A":"60:40 Acetonitrile:Water +10mM Ammonium Formate +10mM Formic Acid","SOLVENT_B":"9:1 Isopropanol:Acetonitrile +10mM Ammonium Formate +10mM Formic Acid","COLUMN_PRESSURE":"450-850 bar","INJECTION_TEMPERATURE":"4 C","INTERNAL_STANDARD":"See data dictionary","RETENTION_TIME":"See data dictionary","SAMPLE_INJECTION":"1.67 uL","ANALYTICAL_TIME":"13 min","CAPILLARY_VOLTAGE":"3500 eV","TIME_PROGRAM":"15 min","WEAK_WASH_SOLVENT_NAME":"Isopropanol","STRONG_WASH_SOLVENT_NAME":"Isopropanol","TARGET_SAMPLE_TEMPERATURE":"Autosampler temp 4 C","RANDOMIZATION_ORDER":"Excel generated"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS","DETECTOR_TYPE":"TOF MCP","SOFTWARE_VERSION":"Masshunter","DATA_FORMAT":".d"},

"MS":{"MS_COMMENTS":"-","INSTRUMENT_NAME":"Agilent 6530 QTOF","INSTRUMENT_TYPE":"QTOF","MS_TYPE":"ESI","ION_MODE":"POSITIVE","CAPILLARY_VOLTAGE":"3500 eV","COLLISION_ENERGY":"25 eV","COLLISION_GAS":"Nitrogen","DRY_GAS_FLOW":"8L/min","DRY_GAS_TEMP":"325 C","FRAGMENT_VOLTAGE":"120 eV","FRAGMENTATION_METHOD":"Auto MS/MS","ION_SOURCE_TEMPERATURE":"325 C","ION_SPRAY_VOLTAGE":"1000","IONIZATION":"Pos","PRECURSOR_TYPE":"Intact Molecule","REAGENT_GAS":"Nitrogen","SOURCE_TEMPERATURE":"325 C","DATAFORMAT":".d","DESOLVATION_GAS_FLOW":"11 L/min","DESOLVATION_TEMPERATURE":"350 C","NEBULIZER":"35 psig","OCTPOLE_VOLTAGE":"750","RESOLUTION_SETTING":"Exteded Dyamic Range","SCAN_RANGE_MOVERZ":"60-1700 Da","SCANNING_CYCLE":"2 Hz","SCANNING_RANGE":"60-1700 Da","SKIMMER_VOLTAGE":"65","MS_RESULTS_FILE":"ST000656_AN001001_Results.txt UNITS:Counts"}

}