{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST000361","ANALYSIS_ID":"AN000593","VERSION":"1","CREATED_ON":"March 11, 2016, 12:19 pm"},

"PROJECT":{"PROJECT_TITLE":"Characterization and Plasticity of the Metabolome in Peripheral Cells in Bipolar I Disorder","PROJECT_TYPE":"Broad Spectrum LCMS","PROJECT_SUMMARY":"Bipolar disorder is a serious condition characterized by severe and debilitating elevations and depressions in mood and energy (1). Bipolar I disorder (BPI) is a particularly severe phenotype which is generally defined as having periods of abnormal moods ranging from “highs,” (mania or hypomania), to “lows” (depression), alternating with periods of relatively normal moods. Key problems in treating bipolar disorder relate to the fact that the pathophysiology of the disorder remains elusive; as a result, there are currently no laboratory tests for diagnosis, to aid treatment decisions, or to predict treatment responses. Recent advances in the study of bipolar disorder increasingly implicate mitochondrial and bioenergetic dysfunction as a common feature of the pathophysiology of the disorder (2). Shifts in metabolism from mitochondrial oxidative phosphorylation toward glycolysis have been observed in the brains of bipolar patients experiencing depressive mood states (3). Increased oxidative stress has also been shown in postmortem brain from bipolar patients (4), and in cultured peripheral fibroblasts from patients with major depression (5). However, little is known about the inherent metabolic state of peripheral cells during bipolar disorder, which may reflect the overall metabolic profile of the individual and may also impact the supply of nutrients to the brain. We propose to characterize the baseline metabolomic profiles of peripheral cells in cultured fibroblasts from patients with BPI and matched non-affected controls. Patients with Bipolar I Disorder (BPI) and matched non-affected controls were recruited. Males and females from all races and ethnicities between the ages of 19-65 participated in the research. Skin biopsies were obtained and fibroblasts were isolated from each of these biopsies. A total of 1 x 〖10〗^7 fibroblasts cells were used for metabolomics. Cell pellets were flash frozen in liquid nitrogen and shipped on dry ice to the NIH Eastern Regional Comprehensive Metabolomic Resource Core at RTI International in North Carolina. Metabolomics will be determined using a broad spectrum metabolomics protocol by RTI.","INSTITUTE":"University of Alabama, Birmingham","DEPARTMENT":"School of Medicine","LABORATORY":"Department of Pathology","LAST_NAME":"Landar","FIRST_NAME":"Aimee","ADDRESS":"BMR2 506 901 9th St South, Birmingham AL 35205","EMAIL":"landar@uab.edu","PHONE":"205-975-9507","FUNDING_SOURCE":"NIH, Common Fund, Pilot and Feasibility"},

"STUDY":{"STUDY_TITLE":"Characterization and Plasticity of the Metabolome in Peripheral Cells in Bipolar I Disorder","STUDY_TYPE":"Broad Spectrum LCMS","STUDY_SUMMARY":"Patients with Bipolar I Disorder (BPI) and matched non-affected controls were recruited. Males and females from all races and ethnicities between the ages of 19-65 participated in the research. Skin biopsies were obtained and fibroblasts were isolated from each of these biopsies. A total of 1 x 〖10〗^7 fibroblasts cells were used for metabolomics. Cell pellets were flash frozen in liquid nitrogen and shipped on dry ice to the NIH Eastern Regional Comprehensive Metabolomic Resource Core at RTI International in North Carolina. Metabolomics will be determined using a broad spectrum metabolomics protocol by RTI","INSTITUTE":"RTI","DEPARTMENT":"Discovery Sciences","LABORATORY":"System and Translational Sciences","LAST_NAME":"Sumner","FIRST_NAME":"Susan","ADDRESS":"3040 Cornwallis Rd, RTP,NC","EMAIL":"ssumner@rti.org","PHONE":"919-541-7479","NUM_GROUPS":"2","TOTAL_SUBJECTS":"18","NUM_MALES":"7","NUM_FEMALES":"11"},

"SUBJECT":{"SUBJECT_TYPE":"human","SUBJECT_SPECIES":"Homo sapiens","AGE_OR_AGE_RANGE":"27-61 years","GENDER":"male/female","HUMAN_RACE":"caucasian, hispanic, african american"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"131",
"Sample ID":"A_0131",
"Factors":{"Group":"Bipolar"},
"Additional sample data":{"Age":"51","Sex":"M","Race":"Hispanic"}
},
{
"Subject ID":"318",
"Sample ID":"B_0318",
"Factors":{"Group":"Control"},
"Additional sample data":{"Age":"61","Sex":"F","Race":"Caucasian"}
},
{
"Subject ID":"255",
"Sample ID":"B_0255",
"Factors":{"Group":"Control"},
"Additional sample data":{"Age":"55","Sex":"M","Race":"African American"}
},
{
"Subject ID":"110",
"Sample ID":"A_0110",
"Factors":{"Group":"Bipolar"},
"Additional sample data":{"Age":"42","Sex":"F","Race":"African-American"}
},
{
"Subject ID":"191",
"Sample ID":"A_0191",
"Factors":{"Group":"Bipolar"},
"Additional sample data":{"Age":"38","Sex":"M","Race":"White"}
},
{
"Subject ID":"61",
"Sample ID":"B_0061",
"Factors":{"Group":"Control"},
"Additional sample data":{"Age":"41","Sex":"F","Race":"African American"}
},
{
"Subject ID":"245",
"Sample ID":"A_0245",
"Factors":{"Group":"Bipolar"},
"Additional sample data":{"Age":"38","Sex":"F","Race":"Hispanic"}
},
{
"Subject ID":"183",
"Sample ID":"B_0183",
"Factors":{"Group":"Control"},
"Additional sample data":{"Age":"27","Sex":"F","Race":"White"}
},
{
"Subject ID":"302",
"Sample ID":"A_0302",
"Factors":{"Group":"Bipolar"},
"Additional sample data":{"Age":"50","Sex":"M","Race":"Caucasian"}
},
{
"Subject ID":"203",
"Sample ID":"B_0203",
"Factors":{"Group":"Control"},
"Additional sample data":{"Age":"28","Sex":"M","Race":"Caucasian"}
},
{
"Subject ID":"182",
"Sample ID":"B_0182",
"Factors":{"Group":"Control"},
"Additional sample data":{"Age":"50","Sex":"F","Race":"White"}
},
{
"Subject ID":"247",
"Sample ID":"B_0247",
"Factors":{"Group":"Control"},
"Additional sample data":{"Age":"49","Sex":"M","Race":"Caucasian"}
},
{
"Subject ID":"293",
"Sample ID":"A_0293",
"Factors":{"Group":"Bipolar"},
"Additional sample data":{"Age":"39","Sex":"F","Race":"African American"}
},
{
"Subject ID":"276",
"Sample ID":"A_0276",
"Factors":{"Group":"Bipolar"},
"Additional sample data":{"Age":"40","Sex":"F","Race":"African-American"}
},
{
"Subject ID":"107",
"Sample ID":"B_0107",
"Factors":{"Group":"Control"},
"Additional sample data":{"Age":"39","Sex":"M","Race":"Caucasian"}
},
{
"Subject ID":"151",
"Sample ID":"B_0151",
"Factors":{"Group":"Control"},
"Additional sample data":{"Age":"55","Sex":"F","Race":"White"}
},
{
"Subject ID":"128",
"Sample ID":"B_0128",
"Factors":{"Group":"Control"},
"Additional sample data":{"Age":"55","Sex":"F","Race":"African-American"}
},
{
"Subject ID":"312",
"Sample ID":"B_0312",
"Factors":{"Group":"Control"},
"Additional sample data":{"Age":"43","Sex":"F","Race":"Hispanic"}
},
{
"Subject ID":"-",
"Sample ID":"Total_Pool_01",
"Factors":{"Group":"-"},
"Additional sample data":{"Age":"-","Sex":"-","Race":"-"}
},
{
"Subject ID":"-",
"Sample ID":"Total_Pool_02",
"Factors":{"Group":"-"},
"Additional sample data":{"Age":"-","Sex":"-","Race":"-"}
},
{
"Subject ID":"-",
"Sample ID":"Total_Pool_03",
"Factors":{"Group":"-"},
"Additional sample data":{"Age":"-","Sex":"-","Race":"-"}
},
{
"Subject ID":"-",
"Sample ID":"Total_Pool_04",
"Factors":{"Group":"-"},
"Additional sample data":{"Age":"-","Sex":"-","Race":"-"}
},
{
"Subject ID":"-",
"Sample ID":"Total_Pool_05",
"Factors":{"Group":"-"},
"Additional sample data":{"Age":"-","Sex":"-","Race":"-"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"For extraction, 500 µL of an ice-cold solution of 0.005 mg/mL Tryptohan-d5 in 90:10 Methanol:Chloroform (v/v) was added to the tubes containing the thawed human dermal fibroblasts cell pellets. MagNA Lyser ceramic beads were added to the tubes, and a MagNA Lyser was used to beat the samples for two thirty seconds pulses at 2,000 rpm, placing the samples on cold block for 5 minutes in between pulses. Sonication for five minutes was performed to the beaten cell samples, followed by centrifugation at room temperature and at 16,000 rcf for 4 minutes. A 300 µL aliquot of each experimental cell sample homogenized supernatant was transferred to new pre-labeled 2.0 mL LoBind Eppendorf tubes and stored at -80 °C for one hour.","SAMPLE_TYPE":"fibroblasts"},

"TREATMENT":{"TREATMENT_SUMMARY":"NA"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Whole study pooled QC samples were created by combining 175 µL aliquot from each of the study samples into a 20 mL scintillation vial. The QC pooled sample was then vortexed for 30 seconds on an analog vortex mixer. Aliquots, 300 µL, were transferred to five new pre-labeled 2.0 mL LoBind Eppendorf tubes and stored at -80 °C for one hour. Next, the supernatants of the study and whole study pooled QC samples, generated above, were dried overnight using a lyophilizer. The residue was reconstituted in 75 µL of reconstitution buffer ( 95:5 Water:Methanol, v/v) and mixed on a multiple tube vortexer for five minutes at 5,000 rpm. Then, the samples were centrifuged at room temperature at 16,000 rcf for four minutes, and the supernatants were transferred to clean pre-labeled autosampler vials for data acquisition."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"UPLC-MS spectra were collected for all samples. UPLC was performed on a Waters Acquity UPLC with an Acquity HSS T3 column (2.1x 100mm x 1.8 µm) at 50 °C using the reversed-phase separation. Water with 0.1% formic acid (mobile phase A) and methanol with 0.1% formic acid (mobile phase B) were used as mobile phases and the metabolites were chromatographically separated using a gradient separation: (see the 3. Sheridan-Mice-Microglia Cells_RP-Metadata and Analytical Metadata.xlsx file for the flow gradient). Mass spectroscopy analysis was performed using a Synapt G2 Si ESI-Q-TOF using a 10 µL injection volume. UPLC-MS data were collected over 70-1000 m/z in both positive and negative modes.","CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Waters Acquity","COLUMN_NAME":"Acquity HSS T3","FLOW_RATE":".4ml/min","METHODS_FILENAME":"RTI-RCMRC-RP","SOLVENT_A":"Water with 0.1% formic acid","SOLVENT_B":"Methanol with 0.1% formic acid","INJECTION_TEMPERATURE":"8 C"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS","OPERATOR_NAME":"Suraj Dhungana","DETECTOR_TYPE":"TOF","SOFTWARE_VERSION":"Mass Lynx 4.1","ACQUISITION_DATE":"8/05/15","ANALYSIS_PROTOCOL_FILE":"RTI-RCMRC-RP-NEG","DATA_FORMAT":"Continuum"},

"MS":{"MS_COMMENTS":"-","INSTRUMENT_NAME":"Waters Synapt G2 Si QTOF","INSTRUMENT_TYPE":"QTOF","MS_TYPE":"ESI","ION_MODE":"NEGATIVE"},

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