{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001213","ANALYSIS_ID":"AN002024","VERSION":"1","CREATED_ON":"July 15, 2019, 3:21 pm"},

"PROJECT":{"PROJECT_TITLE":"Role of 12-LOX in cold adaptation","PROJECT_SUMMARY":"We aimed to study the role of 12-LOX lipid products in the adaptive thermogenesis in mice exposed to cold.","INSTITUTE":"Joslin Diabetes Center - Harvard medical School","LAST_NAME":"Leiria","FIRST_NAME":"Luiz","ADDRESS":"One Joslin Place, 02215, Boston, MA-USA","EMAIL":"luiz.leiria@joslin.harvard.edu","PHONE":"617-309-1967"},

"STUDY":{"STUDY_TITLE":"Serum lipidomic profile of cold-exposed Ucp1cre/12-LOX KO mice","STUDY_SUMMARY":"We aimed to evaluate whether specific deletion of 12-Lipoxygenase (12-LOX) in brown fat can affect the serum concentrations of 12-LOX products under cold exposure.","INSTITUTE":"Joslin Diabetes Center - Harvard medical School","LAST_NAME":"Leiria","FIRST_NAME":"Luiz","ADDRESS":"One Joslin Place, 02215, Boston, MA-USA","EMAIL":"luiz.leiria@joslin.harvard.edu","PHONE":"617-309-1967"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090","GENDER":"Male"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"4621 02",
"Factors":{"Temperature":"22C","Genotype":"KO"}
},
{
"Subject ID":"-",
"Sample ID":"4801 04",
"Factors":{"Temperature":"5C","Genotype":"WT"}
},
{
"Subject ID":"-",
"Sample ID":"4801 01",
"Factors":{"Temperature":"22C","Genotype":"WT"}
},
{
"Subject ID":"-",
"Sample ID":"4761 01",
"Factors":{"Temperature":"5C","Genotype":"KO"}
},
{
"Subject ID":"-",
"Sample ID":"4801 03",
"Factors":{"Temperature":"22C","Genotype":"KO"}
},
{
"Subject ID":"-",
"Sample ID":"4621 04",
"Factors":{"Temperature":"5C","Genotype":"KO"}
},
{
"Subject ID":"-",
"Sample ID":"4671 02",
"Factors":{"Temperature":"22C","Genotype":"WT"}
},
{
"Subject ID":"-",
"Sample ID":"4601 02",
"Factors":{"Temperature":"5C","Genotype":"WT"}
},
{
"Subject ID":"-",
"Sample ID":"4661 01",
"Factors":{"Temperature":"22C","Genotype":"KO"}
},
{
"Subject ID":"-",
"Sample ID":"4801 02",
"Factors":{"Temperature":"22C","Genotype":"WT"}
},
{
"Subject ID":"-",
"Sample ID":"4661 02",
"Factors":{"Temperature":"5C","Genotype":"WT"}
},
{
"Subject ID":"-",
"Sample ID":"4801 05",
"Factors":{"Temperature":"5C","Genotype":"KO"}
},
{
"Subject ID":"-",
"Sample ID":"4621 01",
"Factors":{"Temperature":"5C","Genotype":"WT"}
},
{
"Subject ID":"-",
"Sample ID":"4621 03",
"Factors":{"Temperature":"22C","Genotype":"WT"}
},
{
"Subject ID":"-",
"Sample ID":"4601 03",
"Factors":{"Temperature":"5C","Genotype":"KO"}
},
{
"Subject ID":"-",
"Sample ID":"4831 02",
"Factors":{"Temperature":"22C","Genotype":"KO"}
},
{
"Subject ID":"-",
"Sample ID":"4761 02",
"Factors":{"Temperature":"5C","Genotype":"KO"}
},
{
"Subject ID":"-",
"Sample ID":"4671 01",
"Factors":{"Temperature":"22C","Genotype":"KO"}
},
{
"Subject ID":"-",
"Sample ID":"4601 04",
"Factors":{"Temperature":"22C","Genotype":"WT"}
},
{
"Subject ID":"-",
"Sample ID":"4671 03",
"Factors":{"Temperature":"5C","Genotype":"WT"}
},
{
"Subject ID":"-",
"Sample ID":"4831 01",
"Factors":{"Temperature":"22C","Genotype":"WT"}
},
{
"Subject ID":"-",
"Sample ID":"4841 01",
"Factors":{"Temperature":"5C","Genotype":"WT"}
},
{
"Subject ID":"-",
"Sample ID":"4841 02",
"Factors":{"Temperature":"22C","Genotype":"KO"}
},
{
"Subject ID":"-",
"Sample ID":"4831 03",
"Factors":{"Temperature":"5C","Genotype":"KO"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Mice were anaesthetised with Isoflurane (under controlled flow-rate), and placed in a hot pad, while the blood was drawn from the tail into a tube. The blood was left at 22C for 30 - 60 minutes, and then centrifuged at 14,000 RPM for 3 minutes. The serum fraction was collected and frost at -20C.","SAMPLE_TYPE":"Blood (serum)"},

"TREATMENT":{"TREATMENT_SUMMARY":"Wild-type and Ucp1CRE/12-LOX KO mice were exposed to a short-term cold temperature (1 hour at 5C) or kept at room temperature. After this period, the serum was collected as described. Ucp1CRE/12-LOX KO mice were created through CRISPR-Cas9 technology, as described in Leiria et al., 2019, Cell Metabolism."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Aliquots of 100 µL serum or 1mg protein from homogenized tissue (measured by BCA) were taken, depending on the experiment. 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 31 −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 H 2 O (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:H 2 O (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 LCMS/MS mediator lipidomics platform."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"UHPLC","INSTRUMENT_NAME":"ABI Sciex 5600+ TripleTOF","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. Fullscan 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)."},

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

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}