{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST000659","ANALYSIS_ID":"AN001005","VERSION":"1","CREATED_ON":"June 29, 2017, 11:33 am"},

"PROJECT":{"PROJECT_TITLE":"Changes in metabolites and lipid mediators associated with supervised exercise training for peripheral artery disease","PROJECT_SUMMARY":"Peripheral artery disease (PAD) is a leading cause of cardiovascular related morbidity and mortality, affecting over 8.5 million men and women in the United States and greater than 200 million individuals worldwide. The mainstay of treatment to improve lower limb symptoms is supervised walking therapy, which does not affect plaque morphology or alter conduit artery blood flow, but rather ameliorates endothelial dysfunction, enhances skeletal muscle metabolism and mitochondrial function, and suppresses inflammatory activation. In this pilot feasibility project we will employ metabolic and lipidomic techniques to measure the effects of supervised exercise therapy on primary metabolism, complex lipids, and lipid mediators, and correlate these effects with individual, subject-level measures of the response to exercise therapy among subjects with PAD. The overarching theme of this work is to identify metabolites, complex lipids, and lipid mediators that are associated with the inter-individual variability in the response of subjects with PAD to supervised exercise therapy. This knowledge will significantly enhance our understanding of the pathophysiology of lower extremity symptoms in PAD, as well as the manner in which supervised exercise therapy improves walking intolerance. It will identify novel therapeutic targets and pathways for pharmacologic manipulation in the treatment of PAD. Aside from having the potential to generate multiple high-impact publications, it will serve as the basis for a planned NIH R01 submission by the PI at the conclusion of the award period.","INSTITUTE":"University of Pennsylvania","DEPARTMENT":"Surgery, Perelman School of Medicine","LAST_NAME":"Scott","FIRST_NAME":"Damrauer","ADDRESS":"3400 Spruce Street, 4 Silverstein Philadelphia, PA 19104","EMAIL":"Scott.Damrauer@uphs.upenn.edu","PHONE":"215-615-1698"},

"STUDY":{"STUDY_TITLE":"Changes in metabolites and lipid mediators associated with supervised exercise training for peripheral","STUDY_TYPE":"Timecourse","STUDY_SUMMARY":"Peripheral artery disease (PAD) is a leading cause of cardiovascular related morbidity and mortality, affecting over 8.5 million men and women in the United States and greater than 200 million individuals worldwide. The mainstay of treatment to improve lower limb symptoms is supervised walking therapy, which does not affect plaque morphology or alter conduit artery blood flow, but rather ameliorates endothelial dysfunction, enhances skeletal muscle metabolism and mitochondrial function, and suppresses inflammatory activation. In this pilot feasibility project we will employ metabolic and lipidomic techniques to measure the effects of supervised exercise therapy on primary metabolism, complex lipids, and lipid mediators, and correlate these effects with individual, subject-level measures of the response to exercise therapy among subjects with PAD. The overarching theme of this work is to identify metabolites, complex lipids, and lipid mediators that are associated with the inter-individual variability in the response of subjects with PAD to supervised exercise therapy. This knowledge will significantly enhance our understanding of the pathophysiology of lower extremity symptoms in PAD, as well as the manner in which supervised exercise therapy improves walking intolerance. It will identify novel therapeutic targets and pathways for pharmacologic manipulation in the treatment of PAD. Aside from having the potential to generate multiple high-impact publications, it will serve as the basis for a planned NIH R01 submission by the PI at the conclusion of the award period.","INSTITUTE":"USDA","DEPARTMENT":"Obesity and metabolism research unit","LAST_NAME":"Newman","FIRST_NAME":"John","ADDRESS":"430 West Health Sciences Dr. Davis, Ca, 95616","EMAIL":"john.newman@ars.usda.gov","PHONE":"(530) 752-1009","STUDY_COMMENTS":"Fatty acids measured but not against a calibration curve. Therefore values are expressed as a relative abundance of the entire samples set such that the sum of all subjects eguals 100%."},

"SUBJECT":{"SUBJECT_TYPE":"Human","SUBJECT_SPECIES":"Homo sapiens","TAXONOMY_ID":"9606","GENDER":"Male","SUBJECT_COMMENTS":"Individuals with peripheral arterial disease, as defined by ankle brachial indices ≥0.4 and ≤0.8 and the presence of classic claudication symptoms"},
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{
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"Factors":{"Visit":"v2","Blood Draw":"A"}
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{
"Subject ID":"-",
"Sample ID":"P1 D01",
"Factors":{"Visit":"v3","Blood Draw":"B"}
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{
"Subject ID":"-",
"Sample ID":"P1 E01",
"Factors":{"Visit":"v3","Blood Draw":"A"}
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{
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"Sample ID":"P1 F01",
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"Sample ID":"P1 G01",
"Factors":{"Visit":"v2","Blood Draw":"B"}
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"Sample ID":"P1 H01",
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"Subject ID":"-",
"Sample ID":"P1 A02",
"Factors":{"Visit":"v2","Blood Draw":"B"}
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{
"Subject ID":"-",
"Sample ID":"P1 B02",
"Factors":{"Visit":"v3","Blood Draw":"A"}
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{
"Subject ID":"-",
"Sample ID":"P1 C02",
"Factors":{"Visit":"v2","Blood Draw":"A"}
},
{
"Subject ID":"-",
"Sample ID":"P1 D02",
"Factors":{"Visit":"v3","Blood Draw":"A"}
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{
"Subject ID":"-",
"Sample ID":"P1 E02",
"Factors":{"Visit":"v2","Blood Draw":"B"}
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{
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"Sample ID":"P1 F02",
"Factors":{"Visit":"v2","Blood Draw":"A"}
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"Sample ID":"P1 B04",
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"Sample ID":"P1 C04",
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"Sample ID":"P1 E04",
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"Subject ID":"-",
"Sample ID":"P1 F04",
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{
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{
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{
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{
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"Sample ID":"P1 C06",
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"Sample ID":"P1 H06",
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"Factors":{"Visit":"v3","Blood Draw":"B"}
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{
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"Sample ID":"P1 D07",
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"Sample ID":"P1 E07",
"Factors":{"Visit":"v3","Blood Draw":"A"}
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"Sample ID":"P1 F07",
"Factors":{"Visit":"v2","Blood Draw":"A"}
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{
"Subject ID":"-",
"Sample ID":"P1 G07",
"Factors":{"Visit":"v3","Blood Draw":"A"}
},
{
"Subject ID":"-",
"Sample ID":"P1 H07",
"Factors":{"Visit":"v2","Blood Draw":"A"}
},
{
"Subject ID":"-",
"Sample ID":"P1 A08",
"Factors":{"Visit":"v3","Blood Draw":"A"}
},
{
"Subject ID":"-",
"Sample ID":"P1 B08",
"Factors":{"Visit":"v3","Blood Draw":"A"}
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{
"Subject ID":"-",
"Sample ID":"P1 C08",
"Factors":{"Visit":"v3","Blood Draw":"A"}
},
{
"Subject ID":"-",
"Sample ID":"P1 D08",
"Factors":{"Visit":"v2","Blood Draw":"A"}
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{
"Subject ID":"-",
"Sample ID":"P1 E08",
"Factors":{"Visit":"v3","Blood Draw":"A"}
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{
"Subject ID":"-",
"Sample ID":"P1 F08",
"Factors":{"Visit":"v2","Blood Draw":"B"}
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{
"Subject ID":"-",
"Sample ID":"P1 G08",
"Factors":{"Visit":"v2","Blood Draw":"B"}
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{
"Subject ID":"-",
"Sample ID":"P1 H08",
"Factors":{"Visit":"v3","Blood Draw":"B"}
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{
"Subject ID":"-",
"Sample ID":"P1 A09",
"Factors":{"Visit":"v3","Blood Draw":"A"}
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{
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"Factors":{"Visit":"v2","Blood Draw":"B"}
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"Subject ID":"-",
"Sample ID":"P1 H09",
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{
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"Sample ID":"P1 A10",
"Factors":{"Visit":"v3","Blood Draw":"A"}
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{
"Subject ID":"-",
"Sample ID":"P1 B10",
"Factors":{"Visit":"v3","Blood Draw":"B"}
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{
"Subject ID":"-",
"Sample ID":"P1 C10",
"Factors":{"Visit":"v3","Blood Draw":"A"}
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{
"Subject ID":"-",
"Sample ID":"P1 D10",
"Factors":{"Visit":"v3","Blood Draw":"B"}
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{
"Subject ID":"-",
"Sample ID":"P1 E10",
"Factors":{"Visit":"v3","Blood Draw":"A"}
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{
"Subject ID":"-",
"Sample ID":"P1 F10",
"Factors":{"Visit":"v2","Blood Draw":"B"}
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"Factors":{"Visit":"v2","Blood Draw":"A"}
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{
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"Sample ID":"P1 A11",
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{
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"Factors":{"Visit":"v2","Blood Draw":"B"}
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"Sample ID":"P1 D11",
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{
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"Sample ID":"P1 E11",
"Factors":{"Visit":"v2","Blood Draw":"A"}
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"Sample ID":"P1 F11",
"Factors":{"Visit":"v3","Blood Draw":"B"}
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{
"Subject ID":"-",
"Sample ID":"P1 G11",
"Factors":{"Visit":"v3","Blood Draw":"B"}
},
{
"Subject ID":"-",
"Sample ID":"P1 H11",
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{
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"Sample ID":"P1 A12",
"Factors":{"Visit":"v2","Blood Draw":"A"}
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{
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"Factors":{"Visit":"v2","Blood Draw":"A"}
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{
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{
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"Factors":{"Visit":"v2","Blood Draw":"B"}
},
{
"Subject ID":"-",
"Sample ID":"P2 A01",
"Factors":{"Visit":"v2","Blood Draw":"B"}
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{
"Subject ID":"-",
"Sample ID":"P2 B01",
"Factors":{"Visit":"v3","Blood Draw":"B"}
},
{
"Subject ID":"-",
"Sample ID":"P2 C01",
"Factors":{"Visit":"v2","Blood Draw":"A"}
},
{
"Subject ID":"-",
"Sample ID":"P2 D01",
"Factors":{"Visit":"v3","Blood Draw":"B"}
},
{
"Subject ID":"-",
"Sample ID":"P2 E01",
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{
"Subject ID":"-",
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},
{
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},
{
"Subject ID":"-",
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{
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},
{
"Subject ID":"-",
"Sample ID":"P2 B02",
"Factors":{"Visit":"v2","Blood Draw":"B"}
},
{
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{
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{
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{
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{
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{
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{
"Subject ID":"-",
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},
{
"Subject ID":"-",
"Sample ID":"P2 A04",
"Factors":{"Visit":"v2","Blood Draw":"B"}
},
{
"Subject ID":"-",
"Sample ID":"P2 B04",
"Factors":{"Visit":"v3","Blood Draw":"A"}
},
{
"Subject ID":"-",
"Sample ID":"P2 C04",
"Factors":{"Visit":"v2","Blood Draw":"A"}
},
{
"Subject ID":"-",
"Sample ID":"P2 D04",
"Factors":{"Visit":"v2","Blood Draw":"B"}
},
{
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"Sample ID":"P2 E04",
"Factors":{"Visit":"v3","Blood Draw":"B"}
},
{
"Subject ID":"-",
"Sample ID":"P2 F04",
"Factors":{"Visit":"v2","Blood Draw":"A"}
},
{
"Subject ID":"-",
"Sample ID":"P2 G04",
"Factors":{"Visit":"v3","Blood Draw":"B"}
},
{
"Subject ID":"-",
"Sample ID":"P2 A05",
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},
{
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},
{
"Subject ID":"-",
"Sample ID":"P2 C05",
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},
{
"Subject ID":"-",
"Sample ID":"P2 D05",
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},
{
"Subject ID":"-",
"Sample ID":"P2 E05",
"Factors":{"Visit":"v2","Blood Draw":"B"}
},
{
"Subject ID":"-",
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},
{
"Subject ID":"-",
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},
{
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},
{
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},
{
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"Sample ID":"P2 E06",
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},
{
"Subject ID":"-",
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},
{
"Subject ID":"-",
"Sample ID":"P2 G06",
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},
{
"Subject ID":"-",
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},
{
"Subject ID":"-",
"Sample ID":"P2 A07",
"Factors":{"Visit":"v3","Blood Draw":"B"}
},
{
"Subject ID":"-",
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},
{
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},
{
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},
{
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},
{
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},
{
"Subject ID":"-",
"Sample ID":"P2 G07",
"Factors":{"Visit":"v2","Blood Draw":"B"}
},
{
"Subject ID":"-",
"Sample ID":"P2 H07",
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},
{
"Subject ID":"-",
"Sample ID":"P2 A08",
"Factors":{"Visit":"v2","Blood Draw":"B"}
},
{
"Subject ID":"-",
"Sample ID":"P2 B08",
"Factors":{"Visit":"v3","Blood Draw":"B"}
},
{
"Subject ID":"-",
"Sample ID":"P2 C08",
"Factors":{"Visit":"v2","Blood Draw":"A"}
},
{
"Subject ID":"-",
"Sample ID":"P2 D08",
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{
"Subject ID":"-",
"Sample ID":"P2 E08",
"Factors":{"Visit":"v3","Blood Draw":"A"}
},
{
"Subject ID":"-",
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},
{
"Subject ID":"-",
"Sample ID":"P2 B09",
"Factors":{"Visit":"v3","Blood Draw":"A"}
},
{
"Subject ID":"-",
"Sample ID":"P2 C09",
"Factors":{"Visit":"v2","Blood Draw":"B"}
},
{
"Subject ID":"-",
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},
{
"Subject ID":"-",
"Sample ID":"P2 E09",
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},
{
"Subject ID":"-",
"Sample ID":"P2 F09",
"Factors":{"Visit":"v3","Blood Draw":"A"}
},
{
"Subject ID":"-",
"Sample ID":"P2 G09",
"Factors":{"Visit":"v3","Blood Draw":"B"}
},
{
"Subject ID":"-",
"Sample ID":"P2 H09",
"Factors":{"Visit":"v2","Blood Draw":"B"}
},
{
"Subject ID":"-",
"Sample ID":"P2 A10",
"Factors":{"Visit":"v3","Blood Draw":"A"}
},
{
"Subject ID":"-",
"Sample ID":"P2 B10",
"Factors":{"Visit":"v2","Blood Draw":"B"}
},
{
"Subject ID":"-",
"Sample ID":"P2 C10",
"Factors":{"Visit":"v3","Blood Draw":"A"}
},
{
"Subject ID":"-",
"Sample ID":"P2 D10",
"Factors":{"Visit":"v2","Blood Draw":"B"}
},
{
"Subject ID":"-",
"Sample ID":"P2 E10",
"Factors":{"Visit":"v3","Blood Draw":"B"}
},
{
"Subject ID":"-",
"Sample ID":"P2 F10",
"Factors":{"Visit":"v3","Blood Draw":"A"}
},
{
"Subject ID":"-",
"Sample ID":"P2 G10",
"Factors":{"Visit":"v3","Blood Draw":"A"}
},
{
"Subject ID":"-",
"Sample ID":"P2 H10",
"Factors":{"Visit":"v2","Blood Draw":"A"}
},
{
"Subject ID":"-",
"Sample ID":"P2 A11",
"Factors":{"Visit":"v3","Blood Draw":"A"}
},
{
"Subject ID":"-",
"Sample ID":"P2 B11",
"Factors":{"Visit":"v3","Blood Draw":"B"}
},
{
"Subject ID":"-",
"Sample ID":"P2 C11",
"Factors":{"Visit":"v3","Blood Draw":"B"}
},
{
"Subject ID":"-",
"Sample ID":"P2 D11",
"Factors":{"Visit":"v2","Blood Draw":"A"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Blood was drawn form the subject at the baseline (Visit 2 (V2)) before (B) and after (A) the treadmill exercise test. Further, blood was collected after 12 weeks of supervised walking therapy (Visit 3 (V3)), before (B) and after (A) the treadmill exercise test.","SAMPLE_TYPE":"Plasma"},

"TREATMENT":{"TREATMENT_SUMMARY":"Subjects with peripheral arterial disease, as defined by ankle brachial indices ≥0.4 and ≤0.8 and the presence of classic claudication symptom were subjected to the treadmill exercise test. Further, they underwent 12 weeks of supervised walking therapy and subjected to the treadmill exercise test again."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Lipid mediator extraction and quantification: Oxylipins, endocannabinoids, and fatty acids were isolated from plasma using Ostro Sample Preparation Plates (Waters; Milford, MA) in the presence of BHT/EDTA and quantified by UPLC-MS/MS using internal standard methods. Briefly, in the Ostro Plate well, 100µl plasma aliquots were mixed with 5µl of BHT/EDTA (1:1 v/v MeOH/water) and 5µl methanol containing a suite of deuterated surrogates. The samples were then mixed with 300µL of acetonitrile with 1% formic acid and eluted with 15 mmHg vacuum for 10 min into 200µL glass inserts containing 10µl of 20% glycerol in methanol. Solvent was removed by vacuum centrifugation and the glycerol plug was stored at -80Co until analysis. Prior acquisition, samples were reconstituted in 100µl of 1:1 methanol:acetonitrile containing 100nM of 1-cyclohexyl ureido, 3-dodecanoic acid (CUDA) and 1-phenyl ureido, 3-hexadecanoic acid (PUHA). Residues within extracts were separated on a 2.1 x 150mm 0.17µm BEH column (Waters) and detected by electrospray ionization with multi reaction monitoring on a API 6500 QTRAP (Sciex; Redwood City, CA) and quantified against 7-9 point calibration curves of authentic standards using modifications of previously reported methods (Grapov, D., et al. 2012. PLoS One 7: e48852.; doi: 10.1371/journal.pone.0048852)."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Waters Acquity","COLUMN_NAME":"Waters Acquity BEH C18 (100 x 2mm, 1.7um)","FLOW_GRADIENT":"See protocol/methods file","FLOW_RATE":"0.25 uL","COLUMN_TEMPERATURE":"60 °C","METHODS_FILENAME":"2017_6500_OxyEndo_Analysis_protocol.pdf","SOLVENT_A":"0.1% acetic acid","SOLVENT_B":"90% ACN / 10% IPA","INTERNAL_STANDARD":"See protocol/methods file","RETENTION_TIME":"See protocol/methods file","SAMPLE_INJECTION":"5 µL","ANALYTICAL_TIME":"20 min","WEAK_WASH_SOLVENT_NAME":"20% methanol, 10% isopropanol","WEAK_WASH_VOLUME":"600 µL","STRONG_WASH_SOLVENT_NAME":"50:50 Acetonitrile:Methanol","STRONG_WASH_VOLUME":"600 µL"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS","LABORATORY_NAME":"Newman","OPERATOR_NAME":"Kamil Borkowski","SOFTWARE_VERSION":"AB Sciex MultiQuant version 3.0.1"},

"MS":{"INSTRUMENT_NAME":"ABI Sciex 6500 QTrap","INSTRUMENT_TYPE":"Triple quadrupole","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"Endocannabinoid Analysis"},

"MS_METABOLITE_DATA":{
"Units":"Concentration pmol/g",

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}

}