{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001624","ANALYSIS_ID":"AN002659","VERSION":"1","CREATED_ON":"December 15, 2020, 11:09 am"},

"PROJECT":{"PROJECT_TITLE":"Tissue-specific 1H-NMR metabolomic profiling in mice with adenine-induced chronic kidney disease","PROJECT_TYPE":"Metabolomic profiling of the heart, liver, kidney, and skeletal muscles obtained from mice with and without CKD via 1H NMR","PROJECT_SUMMARY":"This project is focused on a metabolomic analyses of the heart, liver, kidney, and skeletal muscles obtained from mice with and without CKD. To accomplish this objective, we extracted tissues from mice with CKD induced by long-term (24 week) adenine-supplemented diet as well as their control-diet fed counterparts with normal kidney function. Metabolites were extracted from tissues and 1H nuclear magnetic resonance (NMR) was performed and coupled with multivariate statistical analysis.","INSTITUTE":"University of Florida","DEPARTMENT":"Applied Physiology and Kinesiology","LABORATORY":"Rm 42 and Rm 43","LAST_NAME":"Ryan","FIRST_NAME":"Terence","ADDRESS":"1864 Stadium Rd, FLG 114, Gainesville, FL, 32611, USA","EMAIL":"ryant@ufl.edu","PHONE":"352-294-1700","FUNDING_SOURCE":"This research was funded by grants from the National Institutes of Health and the National Heart, Lung, and Blood, Institute numbers R01-HL149704 (to T.E.R.) and the American Heart Association grant number 18CDA34110044 (to T.E.R.).","CONTRIBUTORS":"Ram B. Khattri, Trace Thome, and Terence E. Ryan"},

"STUDY":{"STUDY_TITLE":"Tissue-specific 1H-NMR metabolomic profiling in mice with adenine-induced chronic kidney disease - aqueous phase Kidney (part-I)","STUDY_TYPE":"Metabolomic profiling of the heart, liver, kidney, and skeletal muscles obtained from mice with and without CKD via 1H NMR","STUDY_SUMMARY":"This project is focused on a metabolomic analyses of the heart, liver, kidney, and skeletal muscles obtained from mice with and without CKD. To accomplish this objective, we extracted tissues from mice with CKD induced by long-term (24 week) adenine-supplemented diet as well as their control-diet fed counterparts with normal kidney function. Metabolites were extracted from tissues and 1H nuclear magnetic resonance (NMR) was performed and coupled with multivariate statistical analysis.","INSTITUTE":"University of Florida","DEPARTMENT":"Applied Physiology and Kinesiology","LABORATORY":"Rm 42 and Rm 43","LAST_NAME":"Ryan","FIRST_NAME":"Terence","ADDRESS":"1864 Stadium Rd, FLG 114, Gainesville, FL, 32611, USA","EMAIL":"ryant@ufl.edu","PHONE":"352-294-1700","NUM_GROUPS":"2","TOTAL_SUBJECTS":"17","NUM_MALES":"All","STUDY_COMMENTS":"CKD metabolomic study via NMR using mice model","PUBLICATIONS":"MDPI"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090","GENOTYPE_STRAIN":"C57BL/6J","AGE_OR_AGE_RANGE":"13 months","WEIGHT_OR_WEIGHT_RANGE":"(32.86±1.21 g (control mice) vs 23.57±1.27 g (CKD mice), P < 0.0001, N=7/group).","GENDER":"Male","ANIMAL_ANIMAL_SUPPLIER":"Jackson Labs (Stock # 000664)","ANIMAL_HOUSING":"Housed in a temperature of 22 oC","ANIMAL_LIGHT_CYCLE":"12-hour light/12-hour dark","ANIMAL_FEED":"Ad libitum (Casein control diet vs. adenine-supplemented diet to induce CKD)","ANIMAL_WATER":"free access to food and water (3-5 animals per cage).","ANIMAL_INCLUSION_CRITERIA":"(3-5 animals per cage)."},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"CKD1Kidney_Aq",
"Factors":{"Group":"CKD"},
"Additional sample data":{"RAW_FILE_NAME":"CKD1Kidney_Aq"}
},
{
"Subject ID":"-",
"Sample ID":"CKD2Kidney_Aq",
"Factors":{"Group":"CKD"},
"Additional sample data":{"RAW_FILE_NAME":"CKD2Kidney_Aq"}
},
{
"Subject ID":"-",
"Sample ID":"CKD4Kidney_Aq",
"Factors":{"Group":"CKD"},
"Additional sample data":{"RAW_FILE_NAME":"CKD4Kidney_Aq"}
},
{
"Subject ID":"-",
"Sample ID":"CKD5Kidney_Aq",
"Factors":{"Group":"CKD"},
"Additional sample data":{"RAW_FILE_NAME":"CKD5Kidney_Aq"}
},
{
"Subject ID":"-",
"Sample ID":"CKD6Kidney_Aq",
"Factors":{"Group":"CKD"},
"Additional sample data":{"RAW_FILE_NAME":"CKD6Kidney_Aq"}
},
{
"Subject ID":"-",
"Sample ID":"CKD7Kidney_Aq",
"Factors":{"Group":"CKD"},
"Additional sample data":{"RAW_FILE_NAME":"CKD7Kidney_Aq"}
},
{
"Subject ID":"-",
"Sample ID":"Con1Kidney_Aq",
"Factors":{"Group":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Con1Kidney_Aq"}
},
{
"Subject ID":"-",
"Sample ID":"Con2Kidney_Aq",
"Factors":{"Group":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Con2Kidney_Aq"}
},
{
"Subject ID":"-",
"Sample ID":"Con3Kidney_Aq",
"Factors":{"Group":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Con3Kidney_Aq"}
},
{
"Subject ID":"-",
"Sample ID":"Con4Kidney_Aq",
"Factors":{"Group":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Con4Kidney_Aq"}
},
{
"Subject ID":"-",
"Sample ID":"Con5Kidney_Aq",
"Factors":{"Group":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Con5Kidney_Aq"}
},
{
"Subject ID":"-",
"Sample ID":"Con6Kidney_Aq",
"Factors":{"Group":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Con6Kidney_Aq"}
},
{
"Subject ID":"-",
"Sample ID":"Con7Kidney_Aq",
"Factors":{"Group":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Con7Kidney_Aq"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"While under isoflurane anesthesia, tissues were rapidly dissected and snap frozen in liquid nitrogen and stored at -80°C until metabolite extraction. The following tissues were used in this study: kidney, liver, heart (left ventricle), skeletal muscle (quadriceps). Euthanasia was carried out by thoracotomy followed by cervical dislocation.","SAMPLE_TYPE":"Kidney","COLLECTION_METHOD":"While under isoflurane anesthesia, tissues were rapidly dissected and snap frozen in liquid nitrogen and stored at -80°C until metabolite extraction","COLLECTION_LOCATION":"University of Florida, Applied Physiology and Kinesiology, 1864 stadium RD, Gainesville, FL 32611","STORAGE_CONDITIONS":"-80℃","STORAGE_VIALS":"cryovials"},

"TREATMENT":{"TREATMENT_SUMMARY":"To induce CKD, we utilized an established adenine-diet model. Briefly, mice were assigned to a casein-base chow for 7-days, followed by induction of renal tubular injury by supplementing the diet with 0.2% adenine for 7-days, and subsequently maintained on a 0.15% adenine diet for 5 months and two weeks. CKD mice were then placed back on casein control diet for 2-weeks prior to euthanasia and terminal experiments, allowing for a washout period of adenine based chow. Control mice received casein diet for the duration of the study. Total duration of CKD encompassed 6-months.","ANIMAL_VET_TREATMENTS":"none","ANIMAL_ANESTHESIA":"isoflurane","ANIMAL_FASTING":"non-fasted","ANIMAL_ENDP_EUTHANASIA":"Euthanasia was carried out by thoracotomy followed by cervical dislocation.","ANIMAL_ENDP_TISSUE_COLL_LIST":"kidney, liver, heart (left ventricle), skeletal muscle (quadriceps)"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"A modified form of FOLCH extraction protocol was used to extract metabolites from the tissues. Wet weights of all tissue samples were recorded prior to extraction. Tissue samples were immediately homogenized to prevent any possible enzymatic action using 1 mL of ice-cold methanol in a PowerLyzer 24 Homogenizer (QIAGEN Group, Hilden, Germany). The mixture was centrifuged using 13,200 rpm at 4oC for 30 minutes and the resulting supernatant was transferred to a new glass vial consisting 3 mL of ice cold chloroform:methanol (2:1, v/v) mixture. The homogenate was vortexed and left in an ice bath for 15 minutes to allow for phase separation. Next, 1 mL of 0.9% of saline was added, vortexed it for couple of minutes followed by a second incubation in an ice bath for 30-45 min for complete phase separation. The upper aqueous layer was transferred to a new falcon tube. To the remaining organic phase sample, 1 mL of 0.9% of saline was added again followed by vigorous mixing and letting it stand in ice bath (15 minutes) for a second phase separation. This second aqueous phase was combined with the first. The resulting aqueous and organic layers were dried separately. The aqueous layer was dried overnight with a Labconco freezer dryer (Labconco Corporation, MO, USA) and the organic layer was dried via inert nitrogen gas. These two dried powders (aqueous and organic phases) were stored at -80oC until performing NMR experiments.","PROCESSING_METHOD":"Lyophilization and Homogenization","PROCESSING_STORAGE_CONDITIONS":"-80℃","EXTRACTION_METHOD":"Modified FOLCH extraction","EXTRACT_STORAGE":"-80℃","SAMPLE_RESUSPENSION":"In 50 microliter of 50 mM phosphate buffer (pH 7.2) with 2 mM EDTA, 0.5 mM DSS and 0.2% sodium azide for aqueous phase samples.","SAMPLE_SPIKING":"0.5 mM of DSS for aqueous phase samples"},

"ANALYSIS":{"DATA_FORMAT":"fid, 1r"},

"NM":{"INSTRUMENT_NAME":"Bruker Avance Neo 600 MHz/54mm console","INSTRUMENT_TYPE":"FT-NMR","NMR_EXPERIMENT_TYPE":"1D-1H","FIELD_FREQUENCY_LOCK":"Deuterium","STANDARD_CONCENTRATION":"0.5 mM DSS","SPECTROMETER_FREQUENCY":"600.2328273 MHz","NMR_PROBE":"1.7 mm TXI CryoProbe","NMR_SOLVENT":"Phosphate buffer (pH 7.2) + 2 mM EDTA + 0.5 mM DSS + 0.2% of sodium azide in deuterated environment","NMR_TUBE_SIZE":"1.7 mm O.D.","SHIMMING_METHOD":"Topshim","PULSE_SEQUENCE":"noesypr1d","WATER_SUPPRESSION":"presat","PULSE_WIDTH":"90-degree","RECEIVER_GAIN":"101","OFFSET_FREQUENCY":"2827.31 Hz","CHEMICAL_SHIFT_REF_CPD":"DSS (4,4-dimethyl-4-silapentane-1-sulfonic acid)","TEMPERATURE":"25 o C","NUMBER_OF_SCANS":"128 scans","DUMMY_SCANS":"8","ACQUISITION_TIME":"4 s","RELAXATION_DELAY":"1 s","SPECTRAL_WIDTH":"7142.9 Hz","NUM_DATA_POINTS_ACQUIRED":"28571","REAL_DATA_POINTS":"65536","LINE_BROADENING":"0.22 Hz","ZERO_FILLING":"65,536 points","APODIZATION":"Exponential","BASELINE_CORRECTION_METHOD":"Spline","CHEMICAL_SHIFT_REF_STD":"0 ppm for DSS"},

"NMR_METABOLITE_DATA":{
"Units":"A.U.",

"Data":[{"Metabolite":"Isocaproate","CKD1Kidney_Aq":"9.683602755","CKD2Kidney_Aq":"8.29349982","CKD4Kidney_Aq":"4.745361154","CKD5Kidney_Aq":"13.01075926","CKD6Kidney_Aq":"3.888010368","CKD7Kidney_Aq":"8.009215731","Con1Kidney_Aq":"11.47617311","Con2Kidney_Aq":"20.66492472","Con3Kidney_Aq":"10.69258557","Con4Kidney_Aq":"36.75338604","Con5Kidney_Aq":"24.80784134","Con6Kidney_Aq":"15.98524614","Con7Kidney_Aq":"17.1362385"},{"Metabolite":"Leucine","CKD1Kidney_Aq":"13.27399873","CKD2Kidney_Aq":"11.92590815","CKD4Kidney_Aq":"7.778541898","CKD5Kidney_Aq":"17.50286218","CKD6Kidney_Aq":"8.933183381","CKD7Kidney_Aq":"16.08358924","Con1Kidney_Aq":"17.67675853","Con2Kidney_Aq":"21.43535642","Con3Kidney_Aq":"13.86787943","Con4Kidney_Aq":"23.23210754","Con5Kidney_Aq":"19.9280389","Con6Kidney_Aq":"24.1888152","Con7Kidney_Aq":"24.48005667"},{"Metabolite":"Valine1","CKD1Kidney_Aq":"5.570766552","CKD2Kidney_Aq":"5.152986876","CKD4Kidney_Aq":"3.500566971","CKD5Kidney_Aq":"8.103186209","CKD6Kidney_Aq":"4.244401241","CKD7Kidney_Aq":"6.976735678","Con1Kidney_Aq":"7.214633157","Con2Kidney_Aq":"9.950012641","Con3Kidney_Aq":"5.356281111","Con4Kidney_Aq":"10.39609531","Con5Kidney_Aq":"8.475982685","Con6Kidney_Aq":"10.09514907","Con7Kidney_Aq":"10.65297962"},{"Metabolite":"Isoleucine","CKD1Kidney_Aq":"3.473816323","CKD2Kidney_Aq":"2.952940346","CKD4Kidney_Aq":"2.033218717","CKD5Kidney_Aq":"4.66616518","CKD6Kidney_Aq":"2.149786782","CKD7Kidney_Aq":"4.149502856","Con1Kidney_Aq":"4.338791083","Con2Kidney_Aq":"6.10890184","Con3Kidney_Aq":"3.224872742","Con4Kidney_Aq":"6.996529955","Con5Kidney_Aq":"5.122196882","Con6Kidney_Aq":"5.942119503","Con7Kidney_Aq":"6.259048449"},{"Metabolite":"Valine2","CKD1Kidney_Aq":"5.634644571","CKD2Kidney_Aq":"4.81520252","CKD4Kidney_Aq":"3.303159433","CKD5Kidney_Aq":"7.77174602","CKD6Kidney_Aq":"4.092894274","CKD7Kidney_Aq":"6.759723224","Con1Kidney_Aq":"7.275887308","Con2Kidney_Aq":"9.677509225","Con3Kidney_Aq":"4.883500375","Con4Kidney_Aq":"9.824572676","Con5Kidney_Aq":"8.073518721","Con6Kidney_Aq":"9.31537701","Con7Kidney_Aq":"9.781558473"},{"Metabolite":"Isobutyrate","CKD1Kidney_Aq":"2.644509927","CKD2Kidney_Aq":"1.968583051","CKD4Kidney_Aq":"1.435113157","CKD5Kidney_Aq":"3.036680974","CKD6Kidney_Aq":"1.26665114","CKD7Kidney_Aq":"2.362583144","Con1Kidney_Aq":"2.819925533","Con2Kidney_Aq":"3.455504159","Con3Kidney_Aq":"1.73726236","Con4Kidney_Aq":"5.253228054","Con5Kidney_Aq":"3.64516176","Con6Kidney_Aq":"3.254544358","Con7Kidney_Aq":"3.379622504"},{"Metabolite":"Lactate1","CKD1Kidney_Aq":"156.4193712","CKD2Kidney_Aq":"159.9923236","CKD4Kidney_Aq":"117.8920274","CKD5Kidney_Aq":"282.6286911","CKD6Kidney_Aq":"76.72953474","CKD7Kidney_Aq":"237.0665924","Con1Kidney_Aq":"212.3838656","Con2Kidney_Aq":"261.0791275","Con3Kidney_Aq":"168.4431844","Con4Kidney_Aq":"305.0831008","Con5Kidney_Aq":"295.3134279","Con6Kidney_Aq":"229.9761577","Con7Kidney_Aq":"275.3626179"},{"Metabolite":"Alanine","CKD1Kidney_Aq":"4.867655269","CKD2Kidney_Aq":"2.706745856","CKD4Kidney_Aq":"2.51227014","CKD5Kidney_Aq":"4.855150226","CKD6Kidney_Aq":"1.829170535","CKD7Kidney_Aq":"4.749195172","Con1Kidney_Aq":"6.30061491","Con2Kidney_Aq":"6.359962951","Con3Kidney_Aq":"3.857126708","Con4Kidney_Aq":"6.945433682","Con5Kidney_Aq":"3.500828735","Con6Kidney_Aq":"5.993950544","Con7Kidney_Aq":"6.183183619"},{"Metabolite":"Lysine","CKD1Kidney_Aq":"16.70372335","CKD2Kidney_Aq":"9.984820551","CKD4Kidney_Aq":"9.544000844","CKD5Kidney_Aq":"19.50792879","CKD6Kidney_Aq":"9.397965056","CKD7Kidney_Aq":"20.54354428","Con1Kidney_Aq":"25.18460368","Con2Kidney_Aq":"19.9252455","Con3Kidney_Aq":"16.6758753","Con4Kidney_Aq":"19.26610524","Con5Kidney_Aq":"8.489229574","Con6Kidney_Aq":"27.15080889","Con7Kidney_Aq":"26.96971568"},{"Metabolite":"Acetate","CKD1Kidney_Aq":"7.950489479","CKD2Kidney_Aq":"3.100914646","CKD4Kidney_Aq":"2.527984002","CKD5Kidney_Aq":"6.825978944","CKD6Kidney_Aq":"3.294067886","CKD7Kidney_Aq":"4.457128746","Con1Kidney_Aq":"9.394826741","Con2Kidney_Aq":"5.181422347","Con3Kidney_Aq":"3.2911044","Con4Kidney_Aq":"5.296803041","Con5Kidney_Aq":"4.780470554","Con6Kidney_Aq":"6.040819312","Con7Kidney_Aq":"6.094590145"},{"Metabolite":"Glutamate1","CKD1Kidney_Aq":"25.80188039","CKD2Kidney_Aq":"20.08851544","CKD4Kidney_Aq":"14.99875729","CKD5Kidney_Aq":"47.35464397","CKD6Kidney_Aq":"16.91433033","CKD7Kidney_Aq":"42.17885574","Con1Kidney_Aq":"64.24915306","Con2Kidney_Aq":"55.35192389","Con3Kidney_Aq":"46.54895265","Con4Kidney_Aq":"74.46491209","Con5Kidney_Aq":"51.45839056","Con6Kidney_Aq":"73.23260599","Con7Kidney_Aq":"73.10317601"},{"Metabolite":"Glutamine1+Glutamate+Methionine+O-Acetylcholine","CKD1Kidney_Aq":"40.2136126","CKD2Kidney_Aq":"31.38278353","CKD4Kidney_Aq":"25.49340956","CKD5Kidney_Aq":"74.67037295","CKD6Kidney_Aq":"27.21770642","CKD7Kidney_Aq":"69.79072797","Con1Kidney_Aq":"100.3155406","Con2Kidney_Aq":"79.77522017","Con3Kidney_Aq":"63.26545715","Con4Kidney_Aq":"88.30251637","Con5Kidney_Aq":"69.04225512","Con6Kidney_Aq":"105.6634768","Con7Kid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"Metabolites":[{"metabolite_name":"Isocaproate"},{"metabolite_name":"Leucine"},{"metabolite_name":"Valine1"},{"metabolite_name":"Isoleucine"},{"metabolite_name":"Valine2"},{"metabolite_name":"Isobutyrate"},{"metabolite_name":"Lactate1"},{"metabolite_name":"Alanine"},{"metabolite_name":"Lysine"},{"metabolite_name":"Acetate"},{"metabolite_name":"Glutamate1"},{"metabolite_name":"Glutamine1+Glutamate+Methionine+O-Acetylcholine"},{"metabolite_name":"Glutamate"},{"metabolite_name":"Pyruvate"},{"metabolite_name":"Succinate"},{"metabolite_name":"Citrate-1"},{"metabolite_name":"Aspartate"},{"metabolite_name":"Creatine1"},{"metabolite_name":"Creatinine"},{"metabolite_name":"O-phosphocholine"},{"metabolite_name":"Sn-Glycero-3-Phosphocholine"},{"metabolite_name":"Trimethyl-N-oxide"},{"metabolite_name":"Taurine1"},{"metabolite_name":"Myo-inositol1"},{"metabolite_name":"Taurine-2"},{"metabolite_name":"Myo-Inositol2"},{"metabolite_name":"Glycine"},{"metabolite_name":"Creatine2"},{"metabolite_name":"Myo-inositol3"},{"metabolite_name":"Lactate2"},{"metabolite_name":"Sn-Glycero-3-Phosphocholine2"},{"metabolite_name":"ATP+AMP1"},{"metabolite_name":"Inosine1"},{"metabolite_name":"ATP+AMP2"},{"metabolite_name":"Glucose1"},{"metabolite_name":"Glucose2"},{"metabolite_name":"Allantoin"},{"metabolite_name":"Sucrose"},{"metabolite_name":"Uracil"},{"metabolite_name":"Uridine Triphosphate"},{"metabolite_name":"NAD+1"},{"metabolite_name":"NAD+2"},{"metabolite_name":"ATP+AMP3"},{"metabolite_name":"Fumarate"},{"metabolite_name":"P-Cresol1?"},{"metabolite_name":"p-Cresol2?"},{"metabolite_name":"Tyrosine1"},{"metabolite_name":"Histidine"},{"metabolite_name":"Unknown7.14to7.11ppm"},{"metabolite_name":"Tyrosine2"},{"metabolite_name":"Tryptophan1"},{"metabolite_name":"Phenylalanine"},{"metabolite_name":"Benzoate"},{"metabolite_name":"Nicotinurate2"},{"metabolite_name":"Tyramine?"},{"metabolite_name":"Tryptophan2"},{"metabolite_name":"Xanthine?"},{"metabolite_name":"Unknown7.96to7.94"},{"metabolite_name":"Inosine_or_Adenosine"},{"metabolite_name":"NAD+3"},{"metabolite_name":"Formate"},{"metabolite_name":"NADH"},{"metabolite_name":"NADH2"},{"metabolite_name":"ATP2"},{"metabolite_name":"Unknown8.61to8.59"},{"metabolite_name":"ATP+AMP4"},{"metabolite_name":"Nicotinurate 3"},{"metabolite_name":"NAD+4"},{"metabolite_name":"Nicotinurate4"},{"metabolite_name":"NAD+5"},{"metabolite_name":"NAD+6"}]
}

}