{ "METABOLOMICS WORKBENCH":{"STUDY_ID":"ST000500","ANALYSIS_ID":"AN000769","VERSION":"1","CREATED_ON":"October 31, 2016, 4:41 pm"}, "PROJECT":{"PROJECT_TITLE":"Metabolomics Analysis of Kidney from C57BL/6 Mouse Exposed to Ionizing Radiation","PROJECT_SUMMARY":"NMR based Metabolomics Analysis of Kidney from C57BL/6 Mouse Exposed to Ionizing Radiation","INSTITUTE":"Pacific Northwest National Laboratory","DEPARTMENT":"Fundamental & Computational Sciences","LAST_NAME":"Hu","FIRST_NAME":"Jian Zhi","ADDRESS":"3335 Innovation Blvd, Richland, Washington, 99354, USA","EMAIL":"Jianzhi.Hu@pnnl.gov","PHONE":"(509)371-6544"}, "STUDY":{"STUDY_TITLE":"Metabolomics Analysis of Kidney from C57BL/6 Mouse Exposed to Ionizing Radiation","STUDY_TYPE":"Tissue Extracts Comparison","STUDY_SUMMARY":"Tissue extracts from ionizing readiation exposed mouse kidney and controls were compared via NMR based metabolomic analysis","INSTITUTE":"Pacific Northwest National Laboratory","LAST_NAME":"Hu","FIRST_NAME":"Jian Zhi","ADDRESS":"3335 Innovation Blvd, Richland, Washington, 99354, USA","EMAIL":"Jianzhi.Hu@pnnl.gov","PHONE":"(509)371-6544"}, "SUBJECT":{"SUBJECT_TYPE":"Animal","SUBJECT_SPECIES":"Mus musculus"}, "SUBJECT_SAMPLE_FACTORS":[ { "Subject ID":"-", "Sample ID":"G041", "Factors":{"days post to radiation":"4","radiation dose":"-","radiation source":"sham"} }, { "Subject ID":"-", "Sample ID":"G042", "Factors":{"days post to radiation":"4","radiation dose":"-","radiation source":"sham"} }, { "Subject ID":"-", "Sample ID":"G043", "Factors":{"days post to radiation":"4","radiation dose":"-","radiation source":"sham"} }, { "Subject ID":"-", "Sample ID":"G044", "Factors":{"days post to radiation":"4","radiation dose":"-","radiation source":"sham"} }, { "Subject ID":"-", "Sample ID":"G341", "Factors":{"days post to radiation":"4","radiation dose":"3.0","radiation source":"gamma"} }, { "Subject ID":"-", "Sample ID":"G342", "Factors":{"days post to radiation":"4","radiation dose":"3.0","radiation source":"gamma"} }, { "Subject ID":"-", "Sample ID":"G343", "Factors":{"days post to radiation":"4","radiation dose":"3.0","radiation source":"gamma"} }, { "Subject ID":"-", "Sample ID":"G344", "Factors":{"days post to radiation":"4","radiation dose":"3.0","radiation source":"gamma"} }, { "Subject ID":"-", "Sample ID":"G345", "Factors":{"days post to radiation":"4","radiation dose":"3.0","radiation source":"gamma"} }, { "Subject ID":"-", "Sample ID":"G741", "Factors":{"days post to radiation":"4","radiation dose":"7.8","radiation source":"gamma"} }, { "Subject ID":"-", "Sample ID":"G742", "Factors":{"days post to radiation":"4","radiation dose":"7.8","radiation source":"gamma"} }, { "Subject ID":"-", "Sample ID":"G743", "Factors":{"days post to radiation":"4","radiation dose":"7.8","radiation source":"gamma"} }, { "Subject ID":"-", "Sample ID":"G744", "Factors":{"days post to radiation":"4","radiation dose":"7.8","radiation source":"gamma"} }, { "Subject ID":"-", "Sample ID":"G011", "Factors":{"days post to radiation":"11","radiation dose":"-","radiation source":"sham"} }, { "Subject ID":"-", "Sample ID":"G012", "Factors":{"days post to radiation":"11","radiation dose":"-","radiation source":"sham"} }, { "Subject ID":"-", "Sample ID":"G013", "Factors":{"days post to radiation":"11","radiation dose":"-","radiation source":"sham"} }, { "Subject ID":"-", "Sample ID":"G014", "Factors":{"days post to radiation":"11","radiation dose":"-","radiation source":"sham"} }, { "Subject ID":"-", "Sample ID":"G311", "Factors":{"days post to radiation":"11","radiation dose":"3.0","radiation source":"gamma"} }, { "Subject ID":"-", "Sample ID":"G312", "Factors":{"days post to radiation":"11","radiation dose":"3.0","radiation source":"gamma"} }, { "Subject ID":"-", "Sample ID":"G313", "Factors":{"days post to radiation":"11","radiation dose":"3.0","radiation source":"gamma"} }, { "Subject ID":"-", "Sample ID":"G314", "Factors":{"days post to radiation":"11","radiation dose":"3.0","radiation source":"gamma"} }, { "Subject ID":"-", "Sample ID":"G315", "Factors":{"days post to radiation":"11","radiation dose":"3.0","radiation source":"gamma"} }, { "Subject ID":"-", "Sample ID":"P311", "Factors":{"days post to radiation":"11","radiation dose":"3.0","radiation source":"proton"} }, { "Subject ID":"-", "Sample ID":"P312", "Factors":{"days post to radiation":"11","radiation dose":"3.0","radiation source":"proton"} }, { "Subject ID":"-", "Sample ID":"P313", "Factors":{"days post to radiation":"11","radiation dose":"3.0","radiation source":"proton"} }, { "Subject ID":"-", "Sample ID":"P314", "Factors":{"days post to radiation":"11","radiation dose":"3.0","radiation source":"proton"} }, { "Subject ID":"-", "Sample ID":"P315", "Factors":{"days post to radiation":"11","radiation dose":"3.0","radiation source":"proton"} } ], "COLLECTION":{"COLLECTION_SUMMARY":"Kidneys were dissected and snap-frozen at -80°C before NMR spectroscopy and morphology analysis."}, "TREATMENT":{"TREATMENT_SUMMARY":"A total of 27 seven-week-old C57BL/6 female mice randomly into 6 groups, and then whole body being exposed to either gamma or proton radiation. The individually housed animals were fed a standard diet and maintained in an environment controlled facility with an ambient temperature of 22-25°C and 45% relative humidity on 12 h light-dark cycle. Mice were exposed to whole body gamma radiation using a high-activity source (1250 keV 60Co) with the linear energy transfer (LET) associated with these fields in the range of 0.2-2 keV/μm. The mice were isolated in the corner of their polymer cages and placed a minimum of 100 cm from the collimated 6000 Ci (222 TBq) 60Co source, and then irradiation to the proposed dosage, respectively. The result of absorbed dose rate at a depth of approximately 600 mg/cm2 were ~0.83 Gy/min relative to tissue. After irradiation the isolation barrier was removed and mice were transferred to Animal Facility.Mice placed into aerated polystyrene boxes immediately before exposure to the entrance region of proton at the BNL Alternating Gradient Synchrotron (AGS) (1055 MeV/nucleon at the target, with a track-averaged LET =148.2 keV/μm at target). Doses of 0 and 3.0 Gy were delivered to a maximum of four mice at ~0.83 Gy/min in a single fraction. All procedures (i.e., placement in aerated boxes and subsequent tissue harvesting) for the control mice were identical to those of irradiated animals, except that they received no radiation."}, "SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Step 1: After randomization of the samples, every preweighted intact frozen kidney tissue was homogenized by a Tissue Tearsor in ice-cold after adding 4 ml MeOH and 0.85 ml deionized H2O per gram of liver tissue, followed by vortexing the mixture and then 2 ml chloroform per gram of tissue was added and vortexed again. This process took 6 minutes and kept exactly the same for each sample. Step 2: 2 ml chloroform and 2 ml deionized H2O per gram of tissue were added in the mixture then vortexed again, followed by transferring the different layers into glass vials separately with syringes after the mixture (in ice bath) centrifuged. Finally, the solvents of hydrophilic metabolites were removed by employing lyophilizer and then stored at -80°C freezer before NMR spectral measurements."}, "CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"-","INSTRUMENT_NAME":"-","COLUMN_NAME":"-"}, "ANALYSIS":{"ANALYSIS_TYPE":"NMR"}, "NM":{"INSTRUMENT_NAME":"Varian INOVA 800","INSTRUMENT_TYPE":"FT-NMR","NMR_EXPERIMENT_TYPE":"1D-1H","SPECTROMETER_FREQUENCY":"800MHz"}, "NMR_METABOLITE_DATA":{ "Units":"µM/mg", 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ID":"C00004"},{"Metabolite":"NADP+","PubChem ID":"5886","KEGG ID":"C00006"},{"Metabolite":"NADPH","PubChem ID":"5884","KEGG ID":"C00005"},{"Metabolite":"Niacinamide","PubChem ID":"936","KEGG ID":"C00153"},{"Metabolite":"Nicotinic acid adenine dinucleotide","PubChem ID":"165490","KEGG ID":"C00857"},{"Metabolite":"O-Phosphocholine","PubChem ID":"1014","KEGG ID":"C00588"},{"Metabolite":"O-Phosphoethanolamine","PubChem ID":"1015","KEGG ID":"C00346"},{"Metabolite":"Oxypurinol","PubChem ID":"4644","KEGG ID":"C07599"},{"Metabolite":"Phenylalanine","PubChem ID":"6140","KEGG ID":"C00079"},{"Metabolite":"S-Adenosylhomocysteine","PubChem ID":"439155","KEGG ID":"C00021"},{"Metabolite":"Serine","PubChem ID":"5951","KEGG ID":"C00065"},{"Metabolite":"Succinate","PubChem ID":"1110","KEGG ID":"C00042"},{"Metabolite":"Taurine","PubChem ID":"1123","KEGG ID":"C00245"},{"Metabolite":"Theophylline","PubChem ID":"9340","KEGG ID":"C07130"},{"Metabolite":"Threonine","PubChem ID":"6288","KEGG ID":"C00188"},{"Metabolite":"Thymidine","PubChem ID":"5789","KEGG ID":"C00214"},{"Metabolite":"Tryptophan","PubChem ID":"6305","KEGG ID":"C00078"},{"Metabolite":"Tyrosine","PubChem ID":"3382","KEGG ID":"C00082"},{"Metabolite":"UDP-N-Acetylglucosamine","PubChem ID":"445675","KEGG ID":"C00043"},{"Metabolite":"UDP-galactose","PubChem ID":"18069","KEGG ID":"C00052"},{"Metabolite":"UDP-glucose","PubChem ID":"8629","KEGG ID":"C00029"},{"Metabolite":"UDP-glucuronate","PubChem ID":"17473","KEGG ID":"C00167"},{"Metabolite":"UMP","PubChem ID":"3405","KEGG ID":"C00105"},{"Metabolite":"Uracil","PubChem ID":"1174","KEGG ID":"C00106"},{"Metabolite":"Uridine","PubChem ID":"6029","KEGG ID":"C00299"},{"Metabolite":"Valine","PubChem ID":"6287","KEGG ID":"C00183"},{"Metabolite":"myo-Inositol","PubChem ID":"892","KEGG ID":"C00137"},{"Metabolite":"sn-Glycero-3-phosphocholine","PubChem ID":"439285","KEGG ID":"C00670"},{"Metabolite":"β-Alanine","PubChem ID":"239","KEGG ID":"C00099"},{"Metabolite":"π-Methylhistidine","PubChem ID":"4381","KEGG ID":"C01152"},{"Metabolite":"τ-Methylhistidine","PubChem ID":"92105","KEGG ID":"C01152"}] } }