{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST000401","ANALYSIS_ID":"AN000640","VERSION":"1","CREATED_ON":"May 17, 2016, 3:13 pm"},

"PROJECT":{"PROJECT_TITLE":"Inhibition of diamine oxidase promotes uptake of putrescine from rat small intestine","PROJECT_SUMMARY":"Metformin, a biguanide molecule, which is used as first line therapy for type 2 diabetes. In this study, we would like to investigate the inhibition of an enzyme called diamine oxidase (DAO) (also known as ABP1), by metformin. Based on our preliminary in vitro study using diamine oxidase enzyme, we saw increased level of putrescine with increasing metformin concentrations (see reference PMID: 26335661). This proposed in vivo study was to determine whether metformin could increase putrescine levels and other metabolites in mice. Aminoguanidine, a known inhibitor of DAO, in this study as positive control, following similar study design described in this paper (PMID: 8912017).","INSTITUTE":"University of California, Davis","DEPARTMENT":"Genome and Biomedical Sciences Facility","LABORATORY":"WCMC Metabolomics Core","LAST_NAME":"Fiehn","FIRST_NAME":"Oliver","ADDRESS":"1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616","EMAIL":"ofiehn@ucdavis.edu","PHONE":"(530) 754-8258","FUNDING_SOURCE":"NIH U24DK097154"},

"STUDY":{"STUDY_TITLE":"Inhibition of diamine oxidase promotes uptake of putrescine from rat small intestine","STUDY_SUMMARY":"Metformin, a biguanide molecule, which is used as first line therapy for type 2 diabetes. In this study, we would like to investigate the inhibition of an enzyme called diamine oxidase (DAO) (also known as ABP1), by metformin. Based on our preliminary in vitro study using diamine oxidase enzyme, we saw increased level of putrescine with increasing metformin concentrations (see reference PMID: 26335661). This proposed in vivo study was to determine whether metformin could increase putrescine levels and other metabolites in mice. Aminoguanidine, a known inhibitor of DAO, in this study as positive control, following similar study design described in this paper (PMID: 8912017).","INSTITUTE":"University of California, Davis","DEPARTMENT":"Genome and Biomedical Sciences Facility","LABORATORY":"WCMC Metabolomics Core","LAST_NAME":"Fiehn","FIRST_NAME":"Oliver","ADDRESS":"1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616","EMAIL":"ofiehn@ucdavis.edu","PHONE":"(530) 754-8258","NUM_GROUPS":"5","TOTAL_SUBJECTS":"40"},

"SUBJECT":{"SUBJECT_TYPE":"Animal","SUBJECT_SPECIES":"Rattus norvegicus","TAXONOMY_ID":"10116"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"2B-3-Duo",
"Sample ID":"140603actsa27_1",
"Factors":{"Organ":"Duodenum","Treatment":"Aminoguanidine 10 mg/kg s.c."},
"Additional sample data":{"Tissue Weight (mg)":"212.5"}
},
{
"Subject ID":"2B-4-Duo",
"Sample ID":"140603actsa30_1",
"Factors":{"Organ":"Duodenum","Treatment":"Aminoguanidine 10 mg/kg s.c."},
"Additional sample data":{"Tissue Weight (mg)":"160.5"}
},
{
"Subject ID":"2B-1-Duo",
"Sample ID":"140603actsa06_1",
"Factors":{"Organ":"Duodenum","Treatment":"Aminoguanidine 10 mg/kg s.c."},
"Additional sample data":{"Tissue Weight (mg)":"155.2"}
},
{
"Subject ID":"2B-2-Duo",
"Sample ID":"140603actsa25_1",
"Factors":{"Organ":"Duodenum","Treatment":"Aminoguanidine 10 mg/kg s.c."},
"Additional sample data":{"Tissue Weight (mg)":"123.4"}
},
{
"Subject ID":"2A-4-Duo",
"Sample ID":"140603actsa40_1",
"Factors":{"Organ":"Duodenum","Treatment":"Saline (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"169.5"}
},
{
"Subject ID":"2A-1-Duo",
"Sample ID":"140603actsa31_1",
"Factors":{"Organ":"Duodenum","Treatment":"Saline (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"189.4"}
},
{
"Subject ID":"2A-2-Duo",
"Sample ID":"140603actsa24_1",
"Factors":{"Organ":"Duodenum","Treatment":"Saline (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"125.5"}
},
{
"Subject ID":"2A-3-Duo",
"Sample ID":"140603actsa10_1",
"Factors":{"Organ":"Duodenum","Treatment":"Saline (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"131.2"}
},
{
"Subject ID":"2C-4-Duo",
"Sample ID":"140603actsa29_1",
"Factors":{"Organ":"Duodenum","Treatment":"Metformin 25mg/kg (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"112.7"}
},
{
"Subject ID":"2C-3-Duo",
"Sample ID":"140603actsa33_1",
"Factors":{"Organ":"Duodenum","Treatment":"Metformin 25mg/kg (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"116.1"}
},
{
"Subject ID":"2C-1-Duo",
"Sample ID":"140603actsa18_1",
"Factors":{"Organ":"Duodenum","Treatment":"Metformin 25mg/kg (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"125.6"}
},
{
"Subject ID":"2C-2-Duo",
"Sample ID":"140603actsa20_1",
"Factors":{"Organ":"Duodenum","Treatment":"Metformin 25mg/kg (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"121"}
},
{
"Subject ID":"2E-3-Duo",
"Sample ID":"140603actsa16_1",
"Factors":{"Organ":"Duodenum","Treatment":"Metformin 50mg/kg (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"157.9"}
},
{
"Subject ID":"2E-2-Duo",
"Sample ID":"140603actsa13_1",
"Factors":{"Organ":"Duodenum","Treatment":"Metformin 50mg/kg (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"147.9"}
},
{
"Subject ID":"2E-1-Duo",
"Sample ID":"140603actsa38_1",
"Factors":{"Organ":"Duodenum","Treatment":"Metformin 50mg/kg (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"130.6"}
},
{
"Subject ID":"2E-4-Duo",
"Sample ID":"140603actsa32_3",
"Factors":{"Organ":"Duodenum","Treatment":"Metformin 50mg/kg (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"137.6"}
},
{
"Subject ID":"2D-1-Duo",
"Sample ID":"140603actsa17_1",
"Factors":{"Organ":"Duodenum","Treatment":"Metformin 100mg/kg (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"175.6"}
},
{
"Subject ID":"2D-2-Duo",
"Sample ID":"140603actsa35_1",
"Factors":{"Organ":"Duodenum","Treatment":"Metformin 100mg/kg (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"187.4"}
},
{
"Subject ID":"2D-4-Duo",
"Sample ID":"140603actsa03_1",
"Factors":{"Organ":"Duodenum","Treatment":"Metformin 100mg/kg (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"102.5"}
},
{
"Subject ID":"2D-3-Duo",
"Sample ID":"140603actsa07_1",
"Factors":{"Organ":"Duodenum","Treatment":"Metformin 100mg/kg (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"115.2"}
},
{
"Subject ID":"2B-4-Ile",
"Sample ID":"140603actsa05_1",
"Factors":{"Organ":"Ileum","Treatment":"Aminoguanidine 10 mg/kg s.c."},
"Additional sample data":{"Tissue Weight (mg)":"51.8"}
},
{
"Subject ID":"2B-3-Ile",
"Sample ID":"140603actsa02_1",
"Factors":{"Organ":"Ileum","Treatment":"Aminoguanidine 10 mg/kg s.c."},
"Additional sample data":{"Tissue Weight (mg)":"88.4"}
},
{
"Subject ID":"2B-1-Ile",
"Sample ID":"140603actsa22_1",
"Factors":{"Organ":"Ileum","Treatment":"Aminoguanidine 10 mg/kg s.c."},
"Additional sample data":{"Tissue Weight (mg)":"98.7"}
},
{
"Subject ID":"2B-2-Ile",
"Sample ID":"140603actsa11_1",
"Factors":{"Organ":"Ileum","Treatment":"Aminoguanidine 10 mg/kg s.c."},
"Additional sample data":{"Tissue Weight (mg)":"74.2"}
},
{
"Subject ID":"2A-1-Ile",
"Sample ID":"140603actsa28_1",
"Factors":{"Organ":"Ileum","Treatment":"Saline (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"80.5"}
},
{
"Subject ID":"2A-2-Ile",
"Sample ID":"140603actsa21_1",
"Factors":{"Organ":"Ileum","Treatment":"Saline (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"95.4"}
},
{
"Subject ID":"2A-4-Ile",
"Sample ID":"140603actsa15_1",
"Factors":{"Organ":"Ileum","Treatment":"Saline (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"136.1"}
},
{
"Subject ID":"2A-3-Ile",
"Sample ID":"140603actsa04_1",
"Factors":{"Organ":"Ileum","Treatment":"Saline (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"239"}
},
{
"Subject ID":"2C-1-Ile",
"Sample ID":"140603actsa09_1",
"Factors":{"Organ":"Ileum","Treatment":"Metformin 25mg/kg (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"139.4"}
},
{
"Subject ID":"2C-4-Ile",
"Sample ID":"140603actsa14_1",
"Factors":{"Organ":"Ileum","Treatment":"Metformin 25mg/kg (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"69.4"}
},
{
"Subject ID":"2C-2-Ile",
"Sample ID":"140603actsa01_1",
"Factors":{"Organ":"Ileum","Treatment":"Metformin 25mg/kg (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"79.7"}
},
{
"Subject ID":"2C-3-Ile",
"Sample ID":"140603actsa08_1",
"Factors":{"Organ":"Ileum","Treatment":"Metformin 25mg/kg (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"144.4"}
},
{
"Subject ID":"2E-4-Ile",
"Sample ID":"140603actsa39_1",
"Factors":{"Organ":"Ileum","Treatment":"Metformin 50mg/kg (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"158.6"}
},
{
"Subject ID":"2E-2-Ile",
"Sample ID":"140603actsa34_1",
"Factors":{"Organ":"Ileum","Treatment":"Metformin 50mg/kg (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"91.6"}
},
{
"Subject ID":"2E-3-Ile",
"Sample ID":"140603actsa23_1",
"Factors":{"Organ":"Ileum","Treatment":"Metformin 50mg/kg (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"132.2"}
},
{
"Subject ID":"2E-1-Ile",
"Sample ID":"140603actsa12_1",
"Factors":{"Organ":"Ileum","Treatment":"Metformin 50mg/kg (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"133.7"}
},
{
"Subject ID":"2D-1-Ile",
"Sample ID":"140603actsa26_1",
"Factors":{"Organ":"Ileum","Treatment":"Metformin 100mg/kg (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"94.5"}
},
{
"Subject ID":"2D-2-Ile",
"Sample ID":"140603actsa19_1",
"Factors":{"Organ":"Ileum","Treatment":"Metformin 100mg/kg (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"39.7"}
},
{
"Subject ID":"2D-3-Ile",
"Sample ID":"140603actsa37_1",
"Factors":{"Organ":"Ileum","Treatment":"Metformin 100mg/kg (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"135.5"}
},
{
"Subject ID":"2D-4-Ile",
"Sample ID":"140603actsa36_1",
"Factors":{"Organ":"Ileum","Treatment":"Metformin 100mg/kg (oral administration)"},
"Additional sample data":{"Tissue Weight (mg)":"94.1"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"After 1 hour of treatment, blood was collected from the portal vein. Blood from the portal vein was cannulated with a fine syringe and blood was aspirated. Blood was spun down and plasma was collected and stored in -80. Urine was collected from bladder after 1 hour. Tissues (liver, kidney, intestine) were collected at the end of 1 hour. Only duodenum and ileum were used for metabolomics analysis. This study was conducted under the auspices of approved IACUC procedures in MuriGenics, Inc (Vallejo, CA).","SAMPLE_TYPE":"Tissue","COLLECTION_LOCATION":"MuriGenics, Inc (Vallejo, CA)","COLLECTION_TIME":"1 hour after treatment"},

"TREATMENT":{"TREATMENT_SUMMARY":"Twenty (20) adult rats (male, weighing approximately 200g) were randomized by body weights and assigned to five treatment groups (saline, aminoguanidine (positive control), 3 different oral doses of metformin). Before treatment, rats were starved for 24 hours. On treatment day, saline or aminoguanidine or metformin will be given to the rat.","ANIMAL_FASTING":"24 hours"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"1. Weigh 4 mg tissue sample ( muscle 20 mg) in to a 2.0 ml eppendorf tube. Add 1.0 mL extraction solvent to the tissue sample and using GenoGrinder homogenize samples for 45 seconds ensuring that sample resembles a powder. 2. Centrifuge the samples at 2500 rpm. for 5 minutes. Aliquot 2 X 500µl supernatant, one for analysis and one for a backup sample. Store backup aliquot in the -20°C freezer. 3. Evaporate one 500µl aliquot of the sample in the Labconco Centrivap cold trap concentrator to complete dryness 4. The dried aliquot is then re-suspended with 500l 50% acetonitrile (degassed as given) (only for liver and brain samples). 5. Centrifuge for 2 min at 14000 rcf using the centrifuge Eppendorf 5415. 6. Remove supernatant to a new Eppendorff tube. 7. Evaporate the supernatant to dryness in the the Labconco Centrivap cold trap concentrator. 8. Submit to derivatization.","SAMPLEPREP_PROTOCOL_FILENAME":"SOP_Extraction_of_Mammalian_Tissue_Samples.pdf"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"GC","INSTRUMENT_NAME":"Agilent 6890N","COLUMN_NAME":"Restek Corporation Rtx-5Sil MS","FLOW_RATE":"1 ml/min","COLUMN_TEMPERATURE":"50-330C","METHODS_FILENAME":"Data_Dictionary_Fiehn_laboratory_GCTOF_MS_primary_metabolism_10-15-2013_general.pdf","COLUMN_PRESSURE":"7.7 PSI","INJECTION_TEMPERATURE":"50 C ramped to 250 C by 12 C/s","SAMPLE_INJECTION":"0.5 uL","OVEN_TEMPERATURE":"50°C for 1 min, then ramped at 20°C/min to 330°C, held constant for 5 min","TRANSFERLINE_TEMPERATURE":"230C","WASHING_BUFFER":"Ethyl Acetate","SAMPLE_LOOP_SIZE":"30 m length x 0.25 mm internal diameter","RANDOMIZATION_ORDER":"Excel generated"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS","LABORATORY_NAME":"WCMC Metabolomics Core","DETECTOR_TYPE":"TOF","SOFTWARE_VERSION":"ChromaTOF","DATA_FORMAT":".peg, .txt, .cdf"},

"MS":{"MS_COMMENTS":"-","INSTRUMENT_NAME":"Leco Pegasus III GC TOF","INSTRUMENT_TYPE":"GC-TOF","MS_TYPE":"EI","ION_MODE":"POSITIVE","ION_SOURCE_TEMPERATURE":"250 C","IONIZATION_ENERGY":"70 eV","MASS_ACCURACY":"Nominal","SOURCE_TEMPERATURE":"250 C","SCAN_RANGE_MOVERZ":"85-500 Da","SCANNING_CYCLE":"17 Hz","SCANNING_RANGE":"85-500 Da","SKIMMER_VOLTAGE":"1850 V"},

"MS_METABOLITE_DATA":{
"Units":"counts",

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ID":"5960"},{"Metabolite":"asparagine","Retention Index":"553078","Quantified m/z":"188","KEGG ID":"C00152","PubChem ID":"6267"},{"Metabolite":"ascorbic acid","Retention Index":"672898","Quantified m/z":"332","KEGG ID":"C00072","PubChem ID":"54670067"},{"Metabolite":"arachidonic acid","Retention Index":"834339","Quantified m/z":"91","KEGG ID":"C00219","PubChem ID":"444899"},{"Metabolite":"arachidic acid","Retention Index":"856421","Quantified m/z":"117","KEGG ID":"C06425","PubChem ID":"10467"},{"Metabolite":"aminomalonate","Retention Index":"455754","Quantified m/z":"218","KEGG ID":"C00872","PubChem ID":"100714"},{"Metabolite":"alpha-ketoglutarate","Retention Index":"507392","Quantified m/z":"198","KEGG ID":"C00026","PubChem ID":"51"},{"Metabolite":"allantoic acid","Retention Index":"723539","Quantified m/z":"259","KEGG ID":"C00499","PubChem ID":"203"},{"Metabolite":"alanine-alanine","Retention Index":"522546","Quantified m/z":"116","KEGG ID":"C00993","PubChem 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ID":"92135"},{"Metabolite":"2-monopalmitin","Retention Index":"890356","Quantified m/z":"129","KEGG ID":"","PubChem ID":"123409"},{"Metabolite":"2-ketoisocaproic acid","Retention Index":"290473","Quantified m/z":"89","KEGG ID":"C00233","PubChem ID":"70"},{"Metabolite":"2-hydroxyglutaric acid","Retention Index":"506306","Quantified m/z":"247","KEGG ID":"C02630","PubChem ID":"43"},{"Metabolite":"2-hydroxybutanoic acid","Retention Index":"258161","Quantified m/z":"131","KEGG ID":"C05984","PubChem ID":"440864"},{"Metabolite":"2-deoxytetronic acid NIST","Retention Index":"390517","Quantified m/z":"117","KEGG ID":"","PubChem ID":"192742"},{"Metabolite":"2,5-dihydroxypyrazine NIST","Retention Index":"397526","Quantified m/z":"241","KEGG ID":"","PubChem ID":"23368901"},{"Metabolite":"1-monopalmitin","Retention Index":"901749","Quantified m/z":"129","KEGG ID":"C01885","PubChem ID":"14900"},{"Metabolite":"1,5-anhydroglucitol","Retention Index":"633969","Quantified m/z":"217","KEGG ID":"C07326","PubChem ID":"64960"}]
}

}