#METABOLOMICS WORKBENCH efahy_20230425_000007 DATATRACK_ID:3899 STUDY_ID:ST002589 ANALYSIS_ID:AN004261 PROJECT_ID:PR001020 VERSION 1 CREATED_ON April 25, 2023, 7:56 am #PROJECT PR:PROJECT_TITLE Molecular Transducer of Physical Activity Consortium PR:PROJECT_SUMMARY MoTrPAC is a national research consortium designed to discover and perform PR:PROJECT_SUMMARY preliminary characterization of the range of molecular transducers (the PR:PROJECT_SUMMARY "molecular map") that underlie the effects of physical activity in humans. The PR:PROJECT_SUMMARY program's goal is to study the molecular changes that occur during and after PR:PROJECT_SUMMARY exercise and ultimately to advance the understanding of how physical activity PR:PROJECT_SUMMARY improves and preserves health. Preclinical and clinical studies will examine the PR:PROJECT_SUMMARY systemic effects of endurance and resistance exercise across a range of ages and PR:PROJECT_SUMMARY fitness levels by molecular probing of multiple tissues before and after acute PR:PROJECT_SUMMARY and chronic exercise. This program is the largest targeted NIH investment of PR:PROJECT_SUMMARY funds into the mechanisms of how physical activity improves health and prevents PR:PROJECT_SUMMARY disease. The MoTrPAC program is supported by the NIH Common Fund and is managed PR:PROJECT_SUMMARY by a trans-agency working group representing multiple NIH institutes and PR:PROJECT_SUMMARY centers, led by the NIH Office of Strategic Coordination, National Institute of PR:PROJECT_SUMMARY Arthritis and Musculoskeletal and Skin Diseases, National Institute of Diabetes PR:PROJECT_SUMMARY and Digestive and Kidney Diseases, National Institute on Aging, and National PR:PROJECT_SUMMARY Institute of Biomedical Imaging and Bioengineering. MoTrPAC Steering Committee: PR:PROJECT_SUMMARY Wendy Kohrt, Chair, Russ Tracy, Co-Chair; NIH Program Manager, Concepcion PR:PROJECT_SUMMARY Nierras. Euan Ashley and Matthew Wheeler are the PIs for the Motrpac PR:PROJECT_SUMMARY Bioinformatics / Data Coordination Center. PR:INSTITUTE MoTrPAC PR:LAST_NAME Ashley PR:FIRST_NAME Euan A. PR:ADDRESS 300 Pasteur Dr Rm A265. MC 5319. Stanford CA 94305. USA PR:EMAIL euan@stanford.edu PR:PHONE (650) 498-4900 #STUDY ST:STUDY_TITLE MoTrPAC: Endurance exercise training study in young adult rats, Rat Hippocampus ST:STUDY_TITLE Powder - Targeted Ethanolamides ST:STUDY_SUMMARY The goal of the endurance exercise training study in young adult rats (internal ST:STUDY_SUMMARY code: PASS1B-06) was to perform exercise training studies in adult (6 month) ST:STUDY_SUMMARY F344 rats, and from these rats collect as many tissues as feasible in order to ST:STUDY_SUMMARY provide high quality samples for detailed analysis by chemical analysis sites. ST:STUDY_SUMMARY Tissues were collected from 10-12 rats sedentary control rats concurrent with ST:STUDY_SUMMARY the collection of the 8-week training groups. The 8-week training group and ST:STUDY_SUMMARY controls were from the same cohort and same age at euthanasia (either 8). For ST:STUDY_SUMMARY the older age group, an additional set of controls (n=5-6) were collected with ST:STUDY_SUMMARY the 1-2 week training group. Rats were either sedentary or underwent an exercise ST:STUDY_SUMMARY training program. Rats were exercised on the rodent treadmill 5 days per week ST:STUDY_SUMMARY using a progressive training protocol designed to exercise the rats at ST:STUDY_SUMMARY approximately 70% of VO2max as outlined in the Table on the next page. Training ST:STUDY_SUMMARY was performed no earlier than 10:00 am and no later than 5:00 pm over 5 ST:STUDY_SUMMARY consecutive days per week. Training was initiated with the treadmill set at 70% ST:STUDY_SUMMARY of VO2 max (see tables) and 5 degrees grade for 20 minutes. The duration of ST:STUDY_SUMMARY exercise was increased by one minute each day until day 31 of training (start of ST:STUDY_SUMMARY week 7), when a duration of 50 min was reached. Speed and grade of each training ST:STUDY_SUMMARY session increased in larger increments due to treadmill parameters. The highest ST:STUDY_SUMMARY intensity and duration of training began on day 31. This intensity was ST:STUDY_SUMMARY maintained for the final 10 days of the protocol to ensure steady state had been ST:STUDY_SUMMARY achieved. If any rats were unable to perform at least 4 days of training per ST:STUDY_SUMMARY week they were removed from the study and euthanized. It is important to note ST:STUDY_SUMMARY that the starting treadmill speed varied depending on the sex and age of the ST:STUDY_SUMMARY rat. The initial and maximum speeds were based on VO2max measurements obtained ST:STUDY_SUMMARY during the pre-training testing of the compliant rats. Rats assigned to the ST:STUDY_SUMMARY control group followed a schedule similar to the training group. They were ST:STUDY_SUMMARY placed in one lane on the treadmill for 15 minutes/day, 5 days per week. The ST:STUDY_SUMMARY treadmill was set at 0 m/min at an incline that corresponded to the incline ST:STUDY_SUMMARY being used by the training group. ST:INSTITUTE Emory University ST:DEPARTMENT Biochemistry ST:LABORATORY Integrated Lipidomics and Metabolomics Core ST:LAST_NAME Ortlund ST:FIRST_NAME Eric ST:ADDRESS 1510 Clifton Rd. NE, Room G235, Atlanta, GA 30322 ST:EMAIL eortlun@emory.edu ST:PHONE (404) 727-5014 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Rattus norvegicus SU:TAXONOMY_ID 10116 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - QC_prerun_1 Group:QC-PreRun | Timepoint:0 hour | Sex:- raw_file=MoTrPAC_hippocampus_pass1b_pos_V1-20200828.wiff SUBJECT_SAMPLE_FACTORS - QC_prerun_2 Group:QC-PreRun | Timepoint:0 hour | Sex:- raw_file=MoTrPAC_hippocampus_pass1b_pos_V1-20200828.wiff SUBJECT_SAMPLE_FACTORS - QC_prerun_3 Group:QC-PreRun | Timepoint:0 hour | Sex:- raw_file=MoTrPAC_hippocampus_pass1b_pos_V1-20200828.wiff SUBJECT_SAMPLE_FACTORS - QC_blank_4 Group:QC-Blank | Timepoint:0 hour | Sex:- raw_file=MoTrPAC_hippocampus_pass1b_pos_V1-20200828.wiff SUBJECT_SAMPLE_FACTORS - QC_blank_5 Group:QC-Blank | Timepoint:0 hour | Sex:- raw_file=MoTrPAC_hippocampus_pass1b_pos_V1-20200828.wiff 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SUBJECT_SAMPLE_FACTORS - QC_blank_64 Group:QC-Blank | Timepoint:0 hour | Sex:- raw_file=MoTrPAC_hippocampus_pass1b_pos_V1-20200828.wiff SUBJECT_SAMPLE_FACTORS - QC_Standard_65 Group:QC-ExternalStandard | Timepoint:0 hour | Sex:- raw_file=MoTrPAC_hippocampus_pass1b_pos_V1-20200828.wiff SUBJECT_SAMPLE_FACTORS - QC_Standard_66 Group:QC-ExternalStandard | Timepoint:0 hour | Sex:- raw_file=MoTrPAC_hippocampus_pass1b_pos_V1-20200828.wiff SUBJECT_SAMPLE_FACTORS - QC_Standard_67 Group:QC-ExternalStandard | Timepoint:0 hour | Sex:- raw_file=MoTrPAC_hippocampus_pass1b_pos_V1-20200828.wiff SUBJECT_SAMPLE_FACTORS - QC_Standard_68 Group:QC-ExternalStandard | Timepoint:0 hour | Sex:- raw_file=MoTrPAC_hippocampus_pass1b_pos_V1-20200828.wiff SUBJECT_SAMPLE_FACTORS - QC_Standard_69 Group:QC-ExternalStandard | Timepoint:0 hour | Sex:- raw_file=MoTrPAC_hippocampus_pass1b_pos_V1-20200828.wiff SUBJECT_SAMPLE_FACTORS - QC_Standard_70 Group:QC-ExternalStandard | Timepoint:0 hour | Sex:- raw_file=MoTrPAC_hippocampus_pass1b_pos_V1-20200828.wiff SUBJECT_SAMPLE_FACTORS - QC_Standard_71 Group:QC-ExternalStandard | Timepoint:0 hour | Sex:- raw_file=MoTrPAC_hippocampus_pass1b_pos_V1-20200828.wiff SUBJECT_SAMPLE_FACTORS - QC_blank_72 Group:QC-Blank | Timepoint:0 hour | Sex:- raw_file=MoTrPAC_hippocampus_pass1b_pos_V1-20200828.wiff SUBJECT_SAMPLE_FACTORS - QC_blank_73 Group:QC-Blank | Timepoint:0 hour | Sex:- raw_file=MoTrPAC_hippocampus_pass1b_pos_V1-20200828.wiff #COLLECTION CO:SAMPLE_TYPE Hippocampus #TREATMENT TR:TREATMENT_SUMMARY - #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Purification of oxylipids with SPE column #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Lipids were separated by liquid chromatography with 20 minutes gradient CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME ExionLC AC CH:COLUMN_NAME ThermoScientific Accucore C18 (100 x 4.6mm, 2.6um) CH:SOLVENT_A water with 10 mM ammonium acetate CH:SOLVENT_B acetonitrile with 10 mM ammonium acetate CH:FLOW_GRADIENT t=0 min: 90%/10% (%A/%B) @ 0.5 ml/min | t=0.5 min: 90%/10% (%A/%B) @ 0.5 ml/min CH:FLOW_GRADIENT | t=1 min: 50%/50% (%A/%B) @ 0.5 ml/min | t=2 min: 50%/50% (%A/%B) @ 0.5 ml/min CH:FLOW_GRADIENT | t=2.1 min: 25%/75% (%A/%B) @ 0.5 ml/min | t=5 min: 25%/75% (%A/%B) @ 0.5 CH:FLOW_GRADIENT ml/min | t=7 min: 15%/85% (%A/%B) @ 0.5 ml/min | t=12 min: 15%/85% (%A/%B) @ 0.5 CH:FLOW_GRADIENT ml/min | t=12.1 min: 90%/10% (%A/%B) @ 0.5 ml/min | t=18 min: 90%/10% (%A/%B) @ CH:FLOW_GRADIENT 0.5 ml/min CH:FLOW_RATE 0.5 ml/min CH:COLUMN_TEMPERATURE 50C #ANALYSIS AN:ANALYSIS_TYPE MS AN:ANALYSIS_DETAILS: Peak integration, concentrations calculated base on standards curves equations, AN:ANALYSIS_DETAILS: in the result file data represented in ng/mg of tissue #MS MS:INSTRUMENT_TYPE Triple quadrupole MS:INSTRUMENT_NAME ABI Sciex 5500 Qtrap MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS Multiple reaction monitoring mode (MRM) #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS ng/uL MS_METABOLITE_DATA_START Samples QC_prerun_1 QC_prerun_2 QC_prerun_3 QC_blank_4 QC_blank_5 QC_blank_6 QC_blank_7 90280015208 90412015208 90444015208 90251015208 90265015208 90581015208 90259015208 90252015208 90217015208 90578015208 QC_18 90423015208 90225015208 90229015208 90441015208 90564015208 90267015208 90421015208 90222015208 90426015208 90218015208 QC_29 90567015208 90560015208 90289015208 90449015208 90239015208 90248015208 90450015208 90430015208 90576015208 90232015208 QC_40 90420015208 90559015208 90416015208 90587015208 90422015208 90571015208 90439015208 90223015208 90258015208 90283015208 QC_blank_51 90227015208 90254015208 90410015208 90281015208 90237015208 90585015208 90266015208 90292015208 90245015208 QC_blank_61 QC_blank_62 QC_blank_63 QC_blank_64 QC_Standard_65 QC_Standard_66 QC_Standard_67 QC_Standard_68 QC_Standard_69 QC_Standard_70 QC_Standard_71 QC_blank_72 QC_blank_73 Behenoyl-EA 0.003964098 0.001777509 0.001982117 0.001328546 0.00140703 0.001388422 0.001189621 0.00165798 0.001522529 0.001918558 0.001465876 0.001005079 0.001684589 0.001211289 0.001224305 0.00110254 0.001131339 0.000771535 0.000994431 0.000797598 0.000945083 0.001233467 0.001146472 0.000828021 0.000894815 0.000881347 0.000771666 0.000702123 0.000685624 0.000789377 0.000915307 0.000741946 0.000721268 0.000696236 0.000737412 0.000730355 0.000757761 0.000785333 0.000792944 0.00050765 0.000656013 0.000693396 0.000691249 0.00069926 0.000644885 0.000645644 0.000654437 0.000645713 0.000646149 0.000655944 0.000473721 0.000609586 0.000682871 0.000563807 0.0006719 0.000689724 0.000639844 0.000735583 0.000587168 0.000657529 0.000507216 0.00046222 0.000494372 0.004720179 0.181686047 0.363372093 0.726744186 1.453488372 2.906976744 5.813953488 11.62790698 0.016144202 0 Arachidonoyl-EA 0.000423664 0.00042586 0.00046156 0.000509983 0.000240146 0.000211425 0.000207 0.000291587 0.000261212 0.00055569 0.000235225 0.0002353 0.000426011 0.000190526 0.00020669 0.000317601 0.000169946 7.48393e-05 0.000163579 0.000192004 0.000247213 0.000174812 0.000328729 0.000130113 0.000170778 0.000171516 0.000127875 0.000215049 4.75353e-05 9.2389e-05 0.000238985 0.000187345 0.00012262 0.000161137 0.000167152 0.000228223 0.000202519 0.000140883 0.000209513 5.58783e-05 0.000223223 0.000284731 0.000251436 0.000137353 0.000209119 0.000264711 0.000149865 0.000215756 0.000105734 0.000184942 3.547e-05 0.000200129 0.000158482 0.000150123 0.000145106 0.000344065 0.00015613 9.82673e-05 0.000186576 0.000264549 2.5986e-05 0.00030453 3.1138e-05 0.000247314 0.163517442 0.327034884 0.654069767 1.308139535 2.61627907 5.23255814 10.46511628 0.000398697 0.000422536 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name formula mz neutral_mass refmet_name rt Behenoyl-EA C24H49NO2 384 383.3763 Behenoyl-EA 11.76 Arachidonoyl-EA C22H37NO2 348.2 347.2824 Arachidonoyl-EA 5.59 METABOLITES_END #END