{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST000046","ANALYSIS_ID":"AN000077","VERSION":"1","CREATED_ON":"2016-09-17"},

"PROJECT":{"PROJECT_TITLE":"Identification of Altered Metabolic Pathways in Plasma and CSF in Mild Impairment and Alzheimer’s Disease Using Metabolomics","PROJECT_TYPE":"Untargeted LC-MS Metabolomics","PROJECT_SUMMARY":"Alzheimer’s Disease (AD) currently affects more than 5 million Americans, numbers expected to grow dramatically as the population ages. The changes in AD patients begin decades before the onset of dementia, highlighting urgent need for the development of early diagnostic methods. Compelling data that increased levels of amyloid-beta compromise multiple cellular pathways; the investigation of changes in various cellular networks is essential to our understanding of early disease mechanisms and to identify novel therapeutic We applied a liquid chromatography/mass spectrometry-based non-targeted approach to determine global metabolic changes in plasma and cerebrospinal (CSF) from the same individuals with different AD severity. Metabolic profiling a total of significantly altered 342 plasma and 351 CSF metabolites, of which were identified. Based on the changes of >150 metabolites, we found 23 altered pathways in plasma and 20 in CSF in mild cognitive impairment (MCI) vs. normal (CN) individuals with a false discovery rate <0.05. The number of pathways increased with disease severity in both fluids. Lysine metabolism in and the Krebs cycle in CSF were significantly affected in MCI vs. CN. and sphingolipids transport was altered in both CSF and plasma of AD vs. CN. 30 canonical pathways significantly disturbed in MCI and AD patients included metabolism, Krebs cycle, mitochondrial function, neurotransmitter and amino metabolism, and lipid biosynthesis. Pathways in plasma that discriminated all groups included polyamine, lysine, tryptophan metabolism, and biosynthesis; and in CSF involved cortisone and prostaglandin 2 biosynthesis metabolism. Our data suggest metabolomics could advance our understanding of early disease mechanisms shared in progression from CN to MCI and to AD.","INSTITUTE":"Mayo Clinic","DEPARTMENT":"Neurology","LAST_NAME":"Petersen","FIRST_NAME":"Ronald","ADDRESS":"200 First Street SW, Rochester, MN 55905","EMAIL":"Dasari.Surendra@mayo.edu","PHONE":"507-284-0513","FUNDING_SOURCE":"R01ES020715, AG006786, AG016574","PROJECT_COMMENTS":"Pubmed ID: 23700429"},

"STUDY":{"STUDY_TITLE":"Identification of altered metabolic pathways in Alzheimer's disease, mild impairment and cognitively normals using Metabolomics (plasma)","STUDY_TYPE":"1","STUDY_SUMMARY":"Criteria for the diagnosis of amnestic MCI included: (i) memory complaint by the patient and collateral source; (ii) impairment in 1 or more of the 4 domains (memory, executive functioning/attention, visuospatial, or language); essentially normal functional activities of daily living; and (iv) absence of In general, the amnestic MCI determination is made when the memory measures 1.0â??1.5 SD below the means for age and education appropriate individuals our community; however, rigid cutoffs on psychometric scores were not used to the diagnosis of amnestic MCI which was made on clinical grounds. The diagnosis dementia was made using DSM-IV criteria, and the diagnosis of AD was made using criteria. Subjects were considered to be CN if they performed within the range and did not meet criteria for MCI or dementia.","INSTITUTE":"Mayo Clinic","DEPARTMENT":"Neurology","LAST_NAME":"Petersen","FIRST_NAME":"Ronald","ADDRESS":"-","EMAIL":"Dasari.Surendra@mayo.edu","PHONE":"-","NUM_GROUPS":"1","TOTAL_SUBJECTS":"15"},

"SUBJECT":{"SUBJECT_TYPE":"Human","SUBJECT_SPECIES":"Homo sapiens","TAXONOMY_ID":"9606","SPECIES_GROUP":"Human"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"1",
"Sample ID":"Group1_1",
"Factors":{"Cognitive Status":"AD"}
},
{
"Subject ID":"10",
"Sample ID":"Group1_10",
"Factors":{"Cognitive Status":"AD"}
},
{
"Subject ID":"11",
"Sample ID":"Group1_11",
"Factors":{"Cognitive Status":"AD"}
},
{
"Subject ID":"12",
"Sample ID":"Group1_12",
"Factors":{"Cognitive Status":"AD"}
},
{
"Subject ID":"13",
"Sample ID":"Group1_13",
"Factors":{"Cognitive Status":"AD"}
},
{
"Subject ID":"14",
"Sample ID":"Group1_14",
"Factors":{"Cognitive Status":"AD"}
},
{
"Subject ID":"15",
"Sample ID":"Group1_15",
"Factors":{"Cognitive Status":"AD"}
},
{
"Subject ID":"2",
"Sample ID":"Group1_2",
"Factors":{"Cognitive Status":"AD"}
},
{
"Subject ID":"3",
"Sample ID":"Group1_3",
"Factors":{"Cognitive Status":"AD"}
},
{
"Subject ID":"4",
"Sample ID":"Group1_4",
"Factors":{"Cognitive Status":"AD"}
},
{
"Subject ID":"5",
"Sample ID":"Group1_5",
"Factors":{"Cognitive Status":"AD"}
},
{
"Subject ID":"6",
"Sample ID":"Group1_6",
"Factors":{"Cognitive Status":"AD"}
},
{
"Subject ID":"7",
"Sample ID":"Group1_7",
"Factors":{"Cognitive Status":"AD"}
},
{
"Subject ID":"8",
"Sample ID":"Group1_8",
"Factors":{"Cognitive Status":"AD"}
},
{
"Subject ID":"9",
"Sample ID":"Group1_9",
"Factors":{"Cognitive Status":"AD"}
},
{
"Subject ID":"31",
"Sample ID":"Group3_31",
"Factors":{"Cognitive Status":"CN"}
},
{
"Subject ID":"32",
"Sample ID":"Group3_32",
"Factors":{"Cognitive Status":"CN"}
},
{
"Subject ID":"33",
"Sample ID":"Group3_33",
"Factors":{"Cognitive Status":"CN"}
},
{
"Subject ID":"34",
"Sample ID":"Group3_34",
"Factors":{"Cognitive Status":"CN"}
},
{
"Subject ID":"35",
"Sample ID":"Group3_35",
"Factors":{"Cognitive Status":"CN"}
},
{
"Subject ID":"36",
"Sample ID":"Group3_36",
"Factors":{"Cognitive Status":"CN"}
},
{
"Subject ID":"37",
"Sample ID":"Group3_37",
"Factors":{"Cognitive Status":"CN"}
},
{
"Subject ID":"38",
"Sample ID":"Group3_38",
"Factors":{"Cognitive Status":"CN"}
},
{
"Subject ID":"39",
"Sample ID":"Group3_39",
"Factors":{"Cognitive Status":"CN"}
},
{
"Subject ID":"40",
"Sample ID":"Group3_40",
"Factors":{"Cognitive Status":"CN"}
},
{
"Subject ID":"41",
"Sample ID":"Group3_41",
"Factors":{"Cognitive Status":"CN"}
},
{
"Subject ID":"42",
"Sample ID":"Group3_42",
"Factors":{"Cognitive Status":"CN"}
},
{
"Subject ID":"43",
"Sample ID":"Group3_43",
"Factors":{"Cognitive Status":"CN"}
},
{
"Subject ID":"44",
"Sample ID":"Group3_44",
"Factors":{"Cognitive Status":"CN"}
},
{
"Subject ID":"45",
"Sample ID":"Group3_45",
"Factors":{"Cognitive Status":"CN"}
},
{
"Subject ID":"16",
"Sample ID":"Group2_16",
"Factors":{"Cognitive Status":"MCI"}
},
{
"Subject ID":"17",
"Sample ID":"Group2_17",
"Factors":{"Cognitive Status":"MCI"}
},
{
"Subject ID":"18",
"Sample ID":"Group2_18",
"Factors":{"Cognitive Status":"MCI"}
},
{
"Subject ID":"19",
"Sample ID":"Group2_19",
"Factors":{"Cognitive Status":"MCI"}
},
{
"Subject ID":"20",
"Sample ID":"Group2_20",
"Factors":{"Cognitive Status":"MCI"}
},
{
"Subject ID":"21",
"Sample ID":"Group2_21",
"Factors":{"Cognitive Status":"MCI"}
},
{
"Subject ID":"22",
"Sample ID":"Group2_22",
"Factors":{"Cognitive Status":"MCI"}
},
{
"Subject ID":"23",
"Sample ID":"Group2_23",
"Factors":{"Cognitive Status":"MCI"}
},
{
"Subject ID":"24",
"Sample ID":"Group2_24",
"Factors":{"Cognitive Status":"MCI"}
},
{
"Subject ID":"25",
"Sample ID":"Group2_25",
"Factors":{"Cognitive Status":"MCI"}
},
{
"Subject ID":"26",
"Sample ID":"Group2_26",
"Factors":{"Cognitive Status":"MCI"}
},
{
"Subject ID":"27",
"Sample ID":"Group2_27",
"Factors":{"Cognitive Status":"MCI"}
},
{
"Subject ID":"28",
"Sample ID":"Group2_28",
"Factors":{"Cognitive Status":"MCI"}
},
{
"Subject ID":"29",
"Sample ID":"Group2_29",
"Factors":{"Cognitive Status":"MCI"}
},
{
"Subject ID":"30",
"Sample ID":"Group2_30",
"Factors":{"Cognitive Status":"MCI"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Plasma was collected in EDTA tubes from individuals after an overnight fast standard venipuncture procedures.","SAMPLE_TYPE":"Plasma","COLLECTION_LOCATION":"standard venipuncture"},

"TREATMENT":{"TREATMENT_SUMMARY":"-"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"The metabolite extraction method was performed as described previously [PMID: with minor modifications in the method. Plasma and CSF samples (100 µL) were on ice at 4°C followed by deproteinization with methanol (1:4 ratio of to methanol) and vortexed for 10 s, followed by incubation at â??20°C for 2 Prior to deproteinization, 4 µL of an internal standard solution of (247 ng/µL) was added to each plasma, CSF, and quality control (QC) samples monitor the recovery of extracted metabolites. The samples were centrifuged at g for 20 min at 4°C. The supernatants were lyophilized (Savant, Holbrook, and stored at â??20°C prior to analysis. The samples were reconstituted in buffer and analyzed within 24 hrs."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"HILIC","CHROMATOGRAPHY_TYPE":"HILIC","INSTRUMENT_NAME":"Waters Acquity","COLUMN_NAME":"ethylene-bridged hybrid 2.1 150 mm, 1.7 m; Waters","CHROMATOGRAPHY_COMMENTS":"Metabolite separation in plasma and CSF was achieved using an Acquity UPLC (Waters, Milford, MA) with both hydrophilic interaction chromatography (HILIC) hybrid 2.1150 mm, 1.7 mm; Waters) and reversed-phase liquid chromatography C18 (high-strength silica 2.1150 mm, 1.8 m; Waters). For each column, the run time 20 min at a flow rate of 400 L/min. Reverse-phase chromatography was performed 99% solvent A (5 mmol/L NH4 acetate, 0.1% formic acid, and 1% acetonitrile) to solvent B (95% acetonitrile with 0.1% formic acid). The gradient was 0 min, 0% 1 min, 0% B; 3 min, 5% B; 13.0 min, 100% B; 16 min, 100% B; 16.5 min, 0% B; and min, 0% B. The hydrophilic interaction chromatography gradient was as follows: min, 100% B; 1 min, 100% B; 5 min, 90% B; 13.0 min, 0% B; 16 min, 0% B; 16.5 100% B; and 20 min, 100% B. The injection volume of each sample was 5 L and was maintained at 50C. Each sample was injected and analyzed in duplicate."},

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

"MS":{"INSTRUMENT_NAME":"Agilent 6220 TOF","INSTRUMENT_TYPE":"TOF","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"Positive-ion mode/HILIC: A 6220 ToF MS (Agilent Technologies) was operated in positive and negative electrospray ionization modes using a scan range of m/z. The mass accuracy and mass resolution were <5 parts per million (ppm) and respectively. The instrument settings were as follows: nebulizer gas 325°C, capillary voltage 3.5 kV, capillary temperature 300°C, fragmentor 150 V, skimmer voltage 58 V, octapole voltage 250 V, cycle time 0.5 s, and run 15.0 min."},

"MS_METABOLITE_DATA":{
"Units":"Raw MS Intensities",

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