{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001207","ANALYSIS_ID":"AN002009","VERSION":"1","CREATED_ON":"July 1, 2019, 12:15 pm"},

"PROJECT":{"PROJECT_TITLE":"12-LOX metabolites in cold adaptation","PROJECT_SUMMARY":"The goal of this project is to understand the role of the enzyme 12-lipoxygenase in the adaptive thermogenesis. We found this enzyme is activated by cold stimulation, then producing lipid metabolites in adipose tissue and releasing those into the circulation to regulate fuel utilisation and thermogenic pathways required for the cold adaptation.","INSTITUTE":"Joslin Diabetes Center - Harvard medical School","LABORATORY":"Yu-Hua Tseng lab","LAST_NAME":"Leiria","FIRST_NAME":"Luiz","ADDRESS":"One Joslin Place","EMAIL":"luiz.leiria@joslin.harvard.edu","PHONE":"617-309-1967"},

"STUDY":{"STUDY_TITLE":"Lipidomics in the serum of cold exposed mice treated with 12-LOX inhibitor LOXBlock-1","STUDY_SUMMARY":"We aimed to investigate whether the cold-induced release of 12-LOX products into the circulation were dependent on 12-LOX activation. We pre-treated C57BL6/J mice with the pharmacological inhibitor LOXBlock-1 or its vehicle (DMSO), and after 15 minutes we placed them under cold temperature (5C) for 4 hours. A control group was injected with DMSO and kept at room temperature for the same 4 hours. After this period of time we, collected the blood, and obtained the serum fraction that was immediately frozen and submitted for untargeted lipidomics.","INSTITUTE":"Joslin Diabetes Center- Harvar Medical School","LAST_NAME":"Leiria","FIRST_NAME":"Luiz","ADDRESS":"One Joslin Place, Boston-MA, 02215","EMAIL":"luiz.leiria@joslin.harvard.edu","PHONE":"617-309-1967","NUM_GROUPS":"3"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090","GENOTYPE_STRAIN":"C57BL6/J","AGE_OR_AGE_RANGE":"12 weeks old","WEIGHT_OR_WEIGHT_RANGE":"25-30 g","GENDER":"Male","ANIMAL_ANIMAL_SUPPLIER":"Jax Laboratories","ANIMAL_HOUSING":"Conventional","ANIMAL_LIGHT_CYCLE":"regular 12h light /dark cycles","ANIMAL_FEED":"ad libitum normal chow diet","ANIMAL_WATER":"ad libitum"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"C1",
"Factors":{"Temperature":"5C","Treatment":"DMSO"}
},
{
"Subject ID":"-",
"Sample ID":"C2",
"Factors":{"Temperature":"5C","Treatment":"DMSO"}
},
{
"Subject ID":"-",
"Sample ID":"C3",
"Factors":{"Temperature":"5C","Treatment":"DMSO"}
},
{
"Subject ID":"-",
"Sample ID":"C4",
"Factors":{"Temperature":"5C","Treatment":"DMSO"}
},
{
"Subject ID":"-",
"Sample ID":"C5",
"Factors":{"Temperature":"5C","Treatment":"DMSO"}
},
{
"Subject ID":"-",
"Sample ID":"C6",
"Factors":{"Temperature":"5C","Treatment":"DMSO"}
},
{
"Subject ID":"-",
"Sample ID":"C7",
"Factors":{"Temperature":"5C","Treatment":"DMSO"}
},
{
"Subject ID":"-",
"Sample ID":"LB1",
"Factors":{"Temperature":"5C","Treatment":"LoxBlock-1"}
},
{
"Subject ID":"-",
"Sample ID":"LB2",
"Factors":{"Temperature":"5C","Treatment":"LoxBlock-1"}
},
{
"Subject ID":"-",
"Sample ID":"LB3",
"Factors":{"Temperature":"5C","Treatment":"LoxBlock-1"}
},
{
"Subject ID":"-",
"Sample ID":"LB4",
"Factors":{"Temperature":"5C","Treatment":"LoxBlock-1"}
},
{
"Subject ID":"-",
"Sample ID":"LB5",
"Factors":{"Temperature":"5C","Treatment":"LoxBlock-1"}
},
{
"Subject ID":"-",
"Sample ID":"T1",
"Factors":{"Temperature":"22C","Treatment":"DMSO"}
},
{
"Subject ID":"-",
"Sample ID":"T2",
"Factors":{"Temperature":"22C","Treatment":"DMSO"}
},
{
"Subject ID":"-",
"Sample ID":"T3",
"Factors":{"Temperature":"22C","Treatment":"DMSO"}
},
{
"Subject ID":"-",
"Sample ID":"T4",
"Factors":{"Temperature":"22C","Treatment":"DMSO"}
},
{
"Subject ID":"-",
"Sample ID":"T5",
"Factors":{"Temperature":"22C","Treatment":"DMSO"}
},
{
"Subject ID":"-",
"Sample ID":"T6",
"Factors":{"Temperature":"22C","Treatment":"DMSO"}
},
{
"Subject ID":"-",
"Sample ID":"T7",
"Factors":{"Temperature":"22C","Treatment":"DMSO"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Mice were anaesthetised with Avertine and blood was collected through cardiac puncture. Samples were let in room temperature for 30 minutes, and then centrifuged for 3 minutes at 14,000RPM. Serum fas collected and frozen at -20C for further lipidomics analysis.","SAMPLE_TYPE":"Blood (serum)","STORAGE_CONDITIONS":"-80℃"},

"TREATMENT":{"TREATMENT_SUMMARY":"Mice were injected i.p. 15 minutes before the cold exposure, with 50ul of DMSO (vehicle) or LOXBlock-1. Then, the animas were placed in 5C or 22C temperature for 4 hours, until the samples were collected."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Aliquots of 100 µL serum were taken and a mixture of deuterium-labeled internal standards was added to each aliquot, followed by 3x volume of sample of cold methanol (MeOH). Samples were vortexed for 5 minutes and stored at −20 °C overnight. Cold samples were centrifuged at 14,000g for 10 minutes, and the supernatant was then transferred to a new tube and 3 mL of acidified H2O (pH 3.5) was added to each sample prior to C18 SPE. The methyl formate fractions were collected, dried under nitrogen, and reconstituted in 50 µL MeOH:H2O (1:1, by vol). Samples were transferred to 0.5 mL tubes and centrifuged at 20,000g at 4 °C for 10 minutes. (35ul) of supernatant was transferred to LC–MS/MS vials for analysis using the BERG LC–MS/MS mediator lipidomics platform."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"UHPLC","INSTRUMENT_NAME":"Ekspert MicroLC 200 system","COLUMN_NAME":"Synergi™ Fusion-RP capillary C18 column (150 × 0.5 mm, 4 µm; Phenomenex Inc., Torrance, CA, USA) Inc., Torrance, CA, USA)"},

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

"MS":{"INSTRUMENT_NAME":"ABI Sciex 5600+ TripleTOF","INSTRUMENT_TYPE":"QTOF","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","MS_COMMENTS":"MS spectra were acquired in high-resolution mode (>30,000) using a 100-ms accumulation time per spectrum. Full-scan MS/MS was acquired in high sensitivity mode, with an accumulation time optimized per cycle. Collision energy was set using rolling collision energy with a spread of 15V. The identity of a component was confirmed using PeakView® software (SCIEX), and quantification was performed using MultiQuant™ software (SCIEX). C18SPE cartridges were purchased from Biotage. All solvents were of HPLC or LC-MS/MS grade and were acquired from Sigma-Aldrich, Fisher Scientific, or VWR International."},

"MS_METABOLITE_DATA":{
"Units":"Peak Area",

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