{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST000363","ANALYSIS_ID":"AN000595","VERSION":"1","CREATED_ON":"March 10, 2016, 9:48 pm"},

"PROJECT":{"PROJECT_TITLE":"Metabolic Profiling of Anxiety Prone HSV-Latently Infected Obese Mice","PROJECT_TYPE":"Broad Spectrum LCMS","PROJECT_SUMMARY":"The biological factors that lead children from low socioeconomic backgrounds to be at greater risk for the development of anxiety and learning problems are not well understood. While it is clear that there are genetic components to the risk of developing mental health disorders, a role for environmental factors in inducing these problems has been suggested. Many of these factors that affect the brain likely involve exposures that occur early in life. Two such factors are the higher rates of herpes simplex virus (HSV)-1 seropositivity and prevalence of obesity among these children. HSV-1 infection has been associated with impaired cognition during childhood and mental health problems in adulthood. Additionally, obese adults are shown to have higher HSV-1 titers. Similarly, other studies have correlated a diet high in saturated fat and obesity with increased risk of mood disorders and anxiety. A mouse model of obese HSV-1 latent infection was developed. Broad spectrum metabolomics analysis was performed to better understand the metabolomic profile of hippocampus and to compare this metabolomics profile with that of the hypothalamus, microglia, and peripheral blood mononuclear cells. Brain tissue samples for metabolomics experiments were generated in the following manner: 3-week old mice were placed on a 10% low fat (LF) diet and acclimated for one week prior to intranasal HSV-1 infection or mock infection with PBS. Fourteen days post-infection mice were randomized to either a 45% high fat diet (HF) or remained on the LF diet. Eight weeks post-diet transition, mice were euthanized and brain tissue samples were collected and processed for metabolomics.","INSTITUTE":"University of North Carolina , Chapel Hill","DEPARTMENT":"Gillings School of Public Health","LABORATORY":"Department of Nutrition","LAST_NAME":"Sheridan","FIRST_NAME":"Patricia","ADDRESS":"2002 Hooker Research Center, CB#7461, Chapel Hill NC, 27599","EMAIL":"patricia_sheridan@med.unc.edu","PHONE":"919-843-6434","FUNDING_SOURCE":"NIH, Common Fund, Pilot and Feasibility"},

"STUDY":{"STUDY_TITLE":"Broad Spectrum MS analysis of mouse peripheral blood mononuclear cells (PBMC) from Anxiety Prone HSV-Latently Infected Obese Mice","STUDY_TYPE":"Broad Spectrum LCMS","STUDY_SUMMARY":"Mononuclear cell samples for metabolomics experiments were generated in the following manner: 3-week old mice were placed on a 10% low fat (LF) diet and acclimated for one week prior to intranasal HSV-1 infection or mock infection with PBS. Fourteen days post-infection mice were randomized to either a 45% high fat diet (HF) or remained on the LF diet. Eight weeks post-diet transition, mice were euthanized and mononuclear cell samples were collected and processed for metabolomics.","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":"4","TOTAL_SUBJECTS":"30","NUM_MALES":"30"},

"SUBJECT":{"SUBJECT_TYPE":"mouse","SUBJECT_SPECIES":"Mus musculus","GENOTYPE_STRAIN":"C57Bl/6","AGE_OR_AGE_RANGE":"4.5 mo","WEIGHT_OR_WEIGHT_RANGE":"29.1 g-49.2 g","GENDER":"male","ANIMAL_ANIMAL_SUPPLIER":"JAX","ANIMAL_HOUSING":"4/cage","ANIMAL_FEED":"Purina chow"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"444",
"Sample ID":"Blood444",
"Factors":{"HSV-1":"No","Diet at Week 2":"HF"},
"Additional sample data":{"Weight (g)":"38.1"}
},
{
"Subject ID":"430",
"Sample ID":"Blood430",
"Factors":{"HSV-1":"yes","Diet at Week 2":"LF"},
"Additional sample data":{"Weight (g)":"25.2"}
},
{
"Subject ID":"431",
"Sample ID":"Blood431",
"Factors":{"HSV-1":"no","Diet at Week 2":"LF"},
"Additional sample data":{"Weight (g)":"32.7"}
},
{
"Subject ID":"443",
"Sample ID":"Blood443",
"Factors":{"HSV-1":"No","Diet at Week 2":"HF"},
"Additional sample data":{"Weight (g)":"37.8"}
},
{
"Subject ID":"441",
"Sample ID":"Blood441",
"Factors":{"HSV-1":"yes","Diet at Week 2":"HF"},
"Additional sample data":{"Weight (g)":"38.6"}
},
{
"Subject ID":"428",
"Sample ID":"Blood428",
"Factors":{"HSV-1":"yes","Diet at Week 2":"LF"},
"Additional sample data":{"Weight (g)":"28.9"}
},
{
"Subject ID":"427",
"Sample ID":"Blood427",
"Factors":{"HSV-1":"yes","Diet at Week 2":"HF"},
"Additional sample data":{"Weight (g)":"31.4"}
},
{
"Subject ID":"445",
"Sample ID":"Blood445",
"Factors":{"HSV-1":"No","Diet at Week 2":"HF"},
"Additional sample data":{"Weight (g)":"37.7"}
},
{
"Subject ID":"422",
"Sample ID":"Blood422",
"Factors":{"HSV-1":"no","Diet at Week 2":"LF"},
"Additional sample data":{"Weight (g)":"30.3"}
},
{
"Subject ID":"436",
"Sample ID":"Blood436",
"Factors":{"HSV-1":"yes","Diet at Week 2":"LF"},
"Additional sample data":{"Weight (g)":"29.8"}
},
{
"Subject ID":"442",
"Sample ID":"Blood442",
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},
{
"Subject ID":"418",
"Sample ID":"Blood418",
"Factors":{"HSV-1":"no","Diet at Week 2":"HF"},
"Additional sample data":{"Weight (g)":"30.8"}
},
{
"Subject ID":"433",
"Sample ID":"Blood433",
"Factors":{"HSV-1":"no","Diet at Week 2":"LF"},
"Additional sample data":{"Weight (g)":"26.4"}
},
{
"Subject ID":"439",
"Sample ID":"Blood439",
"Factors":{"HSV-1":"yes","Diet at Week 2":"HF"},
"Additional sample data":{"Weight (g)":"33.8"}
},
{
"Subject ID":"435",
"Sample ID":"Blood435",
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"Additional sample data":{"Weight (g)":"24.9"}
},
{
"Subject ID":"425",
"Sample ID":"Blood425",
"Factors":{"HSV-1":"yes","Diet at Week 2":"HF"},
"Additional sample data":{"Weight (g)":"33.2"}
},
{
"Subject ID":"434",
"Sample ID":"Blood434",
"Factors":{"HSV-1":"no","Diet at Week 2":"LF"},
"Additional sample data":{"Weight (g)":"26.2"}
},
{
"Subject ID":"421",
"Sample ID":"Blood421",
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"Additional sample data":{"Weight (g)":"30.8"}
},
{
"Subject ID":"440",
"Sample ID":"Blood440",
"Factors":{"HSV-1":"yes","Diet at Week 2":"HF"},
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},
{
"Subject ID":"437",
"Sample ID":"Blood437",
"Factors":{"HSV-1":"yes","Diet at Week 2":"LF"},
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},
{
"Subject ID":"420",
"Sample ID":"Blood420",
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"Additional sample data":{"Weight (g)":"33.8"}
},
{
"Subject ID":"423",
"Sample ID":"Blood423",
"Factors":{"HSV-1":"no","Diet at Week 2":"LF"},
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},
{
"Subject ID":"424",
"Sample ID":"Blood424",
"Factors":{"HSV-1":"no","Diet at Week 2":"LF"},
"Additional sample data":{"Weight (g)":"29.8"}
},
{
"Subject ID":"429",
"Sample ID":"Blood429",
"Factors":{"HSV-1":"yes","Diet at Week 2":"LF"},
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},
{
"Subject ID":"419",
"Sample ID":"Blood419",
"Factors":{"HSV-1":"no","Diet at Week 2":"HF"},
"Additional sample data":{"Weight (g)":"33.2"}
},
{
"Subject ID":"432",
"Sample ID":"Blood432",
"Factors":{"HSV-1":"no","Diet at Week 2":"LF"},
"Additional sample data":{"Weight (g)":"29.6"}
},
{
"Subject ID":"438",
"Sample ID":"Blood438",
"Factors":{"HSV-1":"yes","Diet at Week 2":"LF"},
"Additional sample data":{"Weight (g)":"29.9"}
},
{
"Subject ID":"417",
"Sample ID":"Blood417",
"Factors":{"HSV-1":"no","Diet at Week 2":"HF"},
"Additional sample data":{"Weight (g)":"32.6"}
},
{
"Subject ID":"426",
"Sample ID":"Blood426",
"Factors":{"HSV-1":"yes","Diet at Week 2":"HF"},
"Additional sample data":{"Weight (g)":"30.9"}
},
{
"Subject ID":"446",
"Sample ID":"Blood446",
"Factors":{"HSV-1":"No","Diet at Week 2":"HF"},
"Additional sample data":{"Weight (g)":"39"}
},
{
"Subject ID":"-",
"Sample ID":"Total_Pool_01",
"Factors":{"HSV-1":"-","Diet at Week 2":"-"},
"Additional sample data":{"Weight (g)":"-"}
},
{
"Subject ID":"-",
"Sample ID":"Total_Pool_02",
"Factors":{"HSV-1":"-","Diet at Week 2":"-"},
"Additional sample data":{"Weight (g)":"-"}
},
{
"Subject ID":"-",
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"Factors":{"HSV-1":"-","Diet at Week 2":"-"},
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},
{
"Subject ID":"-",
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"Factors":{"HSV-1":"-","Diet at Week 2":"-"},
"Additional sample data":{"Weight (g)":"-"}
},
{
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"Factors":{"HSV-1":"-","Diet at Week 2":"-"},
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}
],
"COLLECTION":{"COLLECTION_SUMMARY":"For extraction, 500 µL of an ice-cold solution of 90:10 Methanol:Chloroform (v/v) ¬was added to the tubes containing the thawed cell pellets. MagNA lyser ceramic beads were added to the tubes, and a MagNA lyser was used to beat the samples for two 30 seconds pulses at 2,000 rpm placing samples on cold block for 5 min in between pulses. Sonication for 5 min was performed to the beaten cell samples followed by centrifugation at room temperature and at 16,000rcf for 4 min. A 350 µL aliquot of each experimental cell sample homogenized supernatant was transferred to a labeled 2.0 mL Lo-Bind Eppendorf tube."},

"TREATMENT":{"TREATMENT_SUMMARY":"3-week old mice were placed on a 10% low fat (LF) diet and acclimated for one week prior to intranasal HSV-1 infection or mock infection with PBS. Fourteen days post-infection mice were randomized to either a 45% high fat diet (HF) or remained on the LF diet. Eight weeks post-diet transition, mice were euthanized and brain tissue samples were collected and processed for metabolomics."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"For extraction, 500 µL of an ice-cold solution of 90:10 Methanol:Chloroform (v/v) ¬was added to the tubes containing the thawed cell pellets. MagNA lyser ceramic beads were added to the tubes, and a MagNA lyser was used to beat the samples for two 30 seconds pulses at 2,000 rpm placing samples on cold block for 5 min in between pulses. Sonication for 5 min was performed to the beaten cell samples followed by centrifugation at room temperature and at 16,000rcf for 4 min. A 350 µL aliquot of each experimental cell sample homogenized supernatant was transferred to a labeled 2.0 mL Lo-Bind Eppendorf tube. Analytical pooled samples were created by combining a 125 µL aliquot from each experimental sample into a labeled 20 mL glass vial. The QC pooled sample was vortexed for 30 sec and 350 µL aliquots were transferred to 5 labeled 2.0 mL Lo-Bind eppendorf tubes. Next, the supernatants of the study and QC pooled samples were dried overnight using a lyophilizer. The residue was reconstituted in 50 µL of reconstitution buffer (95:5 Water:Methanol, v/v) and mixed on an analog vortex mixer for 1 min with speed set at 10. Then, the samples were centrifuged at room temperature for 4 min at 16,000 rcf, and the supernatants were transferred to autosampler vials for data acquisition."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"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:","CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Synapt G2 Si ESI-Q-TOF","COLUMN_NAME":"Acquity HSS T3","METHODS_FILENAME":"RTI-RCMRC-RP"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS","ACQUISITION_DATE":"7/21/15"},

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

"MS_METABOLITE_DATA":{
"Units":"intensity",

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