#METABOLOMICS WORKBENCH stringek_20161208_064337 DATATRACK_ID:796 STUDY_ID:ST000607 ANALYSIS_ID:AN000928 PROJECT_ID:PR000444 VERSION 1 CREATED_ON May 26, 2017, 10:36 am #PROJECT PR:PROJECT_TITLE Metabolomics of Murine Age-related Deep Vein Thrombosis PR:PROJECT_TYPE Quantitative 1D-1H-NMR metabolomics PR:PROJECT_SUMMARY Objective: Age is a significant risk factor for the development of venous PR:PROJECT_SUMMARY thrombosis (VT), but the mechanism(s) that underlie this risk remain(s) PR:PROJECT_SUMMARY undefined and poorly understood. Aging is known to adversely influence PR:PROJECT_SUMMARY inflammation and affect metabolism. Untargeted metabolomics permits an agnostic PR:PROJECT_SUMMARY assessment of the physiological landscape and lends insight into the mechanistic PR:PROJECT_SUMMARY underpinnings of clinical phenotypes. The objective of this exploratory study PR:PROJECT_SUMMARY was to test the feasibility of a metabolomics approach for identifying potential PR:PROJECT_SUMMARY metabolic mechanisms of age-related VT. Methods: We subjected whole blood PR:PROJECT_SUMMARY samples collected from young and old nonthrombosed controls and VT mice 2 days PR:PROJECT_SUMMARY after thrombus induction using the electrolytic inferior vena cava, to a PR:PROJECT_SUMMARY methanol:chloroform extraction and assayed the resulting aqueous fractions using PR:PROJECT_SUMMARY 1D-1H- nuclear magnetic resonance. Normalized mouse metabolite data were PR:PROJECT_SUMMARY compared across groups using analysis of variance (ANOVA) with Holm-Sidak PR:PROJECT_SUMMARY post-testing. In addition, associations between metabolite concentrations and PR:PROJECT_SUMMARY parameters of thrombosis such as thrombus and vein wall weights, and markers of PR:PROJECT_SUMMARY inflammation, vein wall P- and E-selectin levels, were assessed using linear PR:PROJECT_SUMMARY regression. The relatedness of the found significant metabolites was visually PR:PROJECT_SUMMARY assessed using a bioinformatics tool, Metscape, which generates PR:PROJECT_SUMMARY compound-reaction-enzyme-gene networks to aid in the interpretation of PR:PROJECT_SUMMARY metabolomics data. Results: Old mice with VT had a greater mean vein wall weight PR:PROJECT_SUMMARY compared with young mice with VT (P < .05). Clot weight differences between old PR:PROJECT_SUMMARY and young mice followed the same trend as vein wall weight (0.011 % 0.04 g vs PR:PROJECT_SUMMARY 0.008 % 0.003 g; P [ not significant). Glutamine (ANOVA, P < .01), proline PR:PROJECT_SUMMARY (ANOVA, P < .01), and phenylalanine (ANOVA, P <.05) levels were increased in old PR:PROJECT_SUMMARY VT mice compared with age-matched controls and young VT mice. Betaine and/or PR:PROJECT_SUMMARY trimethylamine N-oxide levels were increased in aged mice compared with young PR:PROJECT_SUMMARY animals. Vein wall weight was strongly associated with glutamine (P < .05), and PR:PROJECT_SUMMARY phenylalanine (P <.01) concentrations and there was a trend toward an PR:PROJECT_SUMMARY association with proline (P [ .09) concentration. Vein wall Pselectin, but not PR:PROJECT_SUMMARY E-selectin levels, were increased in old VT mice and were associated with the PR:PROJECT_SUMMARY three found metabolites of age-related VT. Collectively, with the addition of PR:PROJECT_SUMMARY glutamate, these metabolites form a single compound-reaction-enzyme gene network PR:PROJECT_SUMMARY that was generated by Metscape. Conclusions: We used 1D-1H-nuclear magnetic PR:PROJECT_SUMMARY resonance metabolite profiling to identify, for the first time, in an PR:PROJECT_SUMMARY experimental model, three potential metabolites, glutamine, phenylalanine, and PR:PROJECT_SUMMARY proline, associated with age-related VT. These metabolites are metabolically PR:PROJECT_SUMMARY related and their levels are associated with vein wall weight and P-selectin PR:PROJECT_SUMMARY concentrations. In aggregate, these findings provide a “roadmap” of pathways PR:PROJECT_SUMMARY that could be interrogated in future studies, which could include provocation of PR:PROJECT_SUMMARY the glutamine, phenylalanine, and proline pathways in the vein wall. This study PR:PROJECT_SUMMARY introduces metabolomics as a new approach to furthering knowledge about the PR:PROJECT_SUMMARY mechanisms of age-related VT. This study has been published: J Vasc Surg Venous PR:PROJECT_SUMMARY Lymphat Disord. 2016 Apr;4(2):221-30. doi: 10.1016/j.jvsv.2015.09.010. Epub 2015 PR:PROJECT_SUMMARY Nov 24. PR:INSTITUTE University of Michigan PR:DEPARTMENT Jobst Vascular Research Laboratories, Section of Vascular Surgery, Department of PR:DEPARTMENT Surgery, School of Medicine; Clinical Pharmacy, College of Pharmacy; Unit for PR:DEPARTMENT Laboratory Animal Medicine; University of Michigan PR:LABORATORY NMR Metabolomics Laboratory, University of Michigan PR:LAST_NAME Stringer PR:FIRST_NAME Kathleen PR:ADDRESS College of Pharmacy, University of Michigan, 428 Church Street, Ann Arbor, MI PR:ADDRESS 48109 PR:EMAIL stringek@umich.edu PR:PHONE NA PR:FUNDING_SOURCE NIH (DK097153; HL076123), College of Pharmacy Upjohn Award #STUDY ST:STUDY_TITLE 1D-1H-Nuclear Magnetic Resonance Metabolomics Reveals Age-related Changes in ST:STUDY_TITLE Metabolites Associated with Experimental Venous Thrombosis ST:STUDY_TYPE Experimental venous thrombosis ST:STUDY_SUMMARY Sodium heparin preserved whole blood samples were collected from young and old ST:STUDY_SUMMARY mice with and without induced VT. Samples were subjected to MeOH:CHCl3 ST:STUDY_SUMMARY extraction and the MeOH was assayed by 1H-NMR. Chenomx software was used for ST:STUDY_SUMMARY spectral analysis. ST:INSTITUTE University of Michigan ST:DEPARTMENT Clinical Pharmacy ST:LABORATORY The NMR Metabolomics Laboratory (Stringer) ST:LAST_NAME Stringer ST:FIRST_NAME Kathleen ST:ADDRESS College of Pharmacy, University of Michigan, 428 Church Street, Ann Arbor, MI ST:ADDRESS 48109 ST:EMAIL stringek@umich.edu ST:PHONE NA ST:NUM_GROUPS 4 ST:TOTAL_SUBJECTS 31 #SUBJECT SU:SUBJECT_TYPE Animal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENOTYPE_STRAIN C57Bl6 SU:AGE_OR_AGE_RANGE 10 weeks and 12-24 months (young & old) SU:WEIGHT_OR_WEIGHT_RANGE ~25 g and ~34 g SU:GENDER Male #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - CH1WB Condition:DVT young Gender=Male SUBJECT_SAMPLE_FACTORS - CH4WB Condition:DVT young Gender=Male SUBJECT_SAMPLE_FACTORS - CH5WB Condition:DVT young Gender=Male SUBJECT_SAMPLE_FACTORS - CH22WB Condition:DVT young Gender=Male SUBJECT_SAMPLE_FACTORS - CH24WB Condition:DVT young Gender=Male SUBJECT_SAMPLE_FACTORS - CH25WB Condition:DVT young Gender=Male SUBJECT_SAMPLE_FACTORS - CH26WB Condition:DVT young Gender=Male SUBJECT_SAMPLE_FACTORS - CH7WB Condition:DVT old Gender=Male SUBJECT_SAMPLE_FACTORS - CH8WB Condition:DVT old Gender=Male SUBJECT_SAMPLE_FACTORS - CH10WB Condition:DVT old Gender=Male SUBJECT_SAMPLE_FACTORS - CH21WB Condition:DVT old Gender=Male SUBJECT_SAMPLE_FACTORS - CH27WB Condition:DVT old Gender=Male SUBJECT_SAMPLE_FACTORS - CH28WB Condition:DVT old Gender=Male SUBJECT_SAMPLE_FACTORS - CH29WB Condition:DVT old Gender=Male SUBJECT_SAMPLE_FACTORS - CH31WB Condition:DVT old Gender=Male SUBJECT_SAMPLE_FACTORS - CH11WB Condition:Cntrl young Gender=Male SUBJECT_SAMPLE_FACTORS - CH12WB Condition:Cntrl young Gender=Male SUBJECT_SAMPLE_FACTORS - CH13WB Condition:Cntrl young Gender=Male SUBJECT_SAMPLE_FACTORS - CH14WB Condition:Cntrl young Gender=Male SUBJECT_SAMPLE_FACTORS - CH15WB Condition:Cntrl young Gender=Male SUBJECT_SAMPLE_FACTORS - CH16WB Condition:Cntrol old Gender=Male SUBJECT_SAMPLE_FACTORS - CH17WB Condition:Cntrol old Gender=Male SUBJECT_SAMPLE_FACTORS - CH18WB Condition:Cntrol old Gender=Male SUBJECT_SAMPLE_FACTORS - CH19WB Condition:Cntrol old Gender=Male SUBJECT_SAMPLE_FACTORS - CH20WB Condition:Cntrol old Gender=Male #COLLECTION CO:COLLECTION_SUMMARY None CO:COLLECTION_PROTOCOL_FILENAME SOP_for_whole_blood_collection_exp._animals_v1.3.pdf CO:SAMPLE_TYPE Whole blood CO:COLLECTION_TUBE_TEMP 4C CO:ADDITIVES Sodium heparin CO:BLOOD_SERUM_OR_PLASMA Whole blood #TREATMENT TR:TREATMENT_SUMMARY None TR:ANIMAL_VET_TREATMENTS Electrolytic inferior vena cava (IVC) model (EIM to induce thrombogenesis TR:ANIMAL_ANESTHESIA 2-5% isoflurane with 100% oxygen TR:ANIMAL_FASTING No TR:ANIMAL_ENDP_EUTHANASIA Per IUCUCA and 2d following induction of VT TR:ANIMAL_ENDP_TISSUE_COLL_LIST Blood sampling, thrombus and vein wall #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Methanol:cholorform extraction SP:SAMPLEPREP_PROTOCOL_FILENAME Wakefield Mouse WB_Extraction Protocol.pdf SP:SAMPLEPREP_PROTOCOL_COMMENTS Lipid fractions saved, stored at -20°C SP:PROCESSING_METHOD Methanol:Chloroform Extraction SP:PROCESSING_STORAGE_CONDITIONS 0-4°C SP:EXTRACTION_METHOD Methanol:chloroform extraction SP:EXTRACT_CLEANUP N/A SP:EXTRACT_STORAGE -80°C #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE - CH:INSTRUMENT_NAME - CH:COLUMN_NAME - #ANALYSIS AN:ANALYSIS_TYPE NMR #NMR NM:INSTRUMENT_NAME Agilent 500/54 Premium Shielded VNMRS system NM:INSTRUMENT_TYPE FT-NMR NM:NMR_EXPERIMENT_TYPE 1D-1H NM:STANDARD_CONCENTRATION 10% (~0.5mM) NM:SPECTROMETER_FREQUENCY 500 MHz NM:NMR_PROBE ONE-Probe NM:NMR_SOLVENT D2O NM:NMR_TUBE_SIZE 5mm NM:SHIMMING_METHOD Auto shim (gradient shimming) NM:PULSE_SEQUENCE 1D-NOESY NM:WATER_SUPPRESSION saturation at 80 Hz induced field strength NM:PULSE_WIDTH 5.5ms NM:OFFSET_FREQUENCY around -178Hz NM:CHEMICAL_SHIFT_REF_CPD 6mM Formate NM:TEMPERATURE 25°C NM:NUMBER_OF_SCANS 32 #NMR_METABOLITE_DATA NMR_METABOLITE_DATA:UNITS uM NMR_METABOLITE_DATA_START Samples CH1WB CH4WB CH5WB CH22WB CH24WB CH25WB CH26WB CH7WB CH8WB CH10WB CH21WB CH27WB CH28WB CH29WB CH31WB CH11WB CH12WB CH13WB CH14WB CH15WB CH16WB CH17WB CH18WB CH19WB CH20WB Factors Condition:DVT young Condition:DVT young Condition:DVT young Condition:DVT young Condition:DVT young Condition:DVT young Condition:DVT young Condition:DVT old Condition:DVT old Condition:DVT old Condition:DVT old Condition:DVT old Condition:DVT old Condition:DVT old Condition:DVT old Condition:Cntrl young Condition:Cntrl young Condition:Cntrl young Condition:Cntrl young Condition:Cntrl young Condition:Cntrol old Condition:Cntrol old Condition:Cntrol old Condition:Cntrol old Condition:Cntrol old 2-Hydroxybutyrate 6.3 8.9 21 6.9 20.7 18.5 18 18 8.8 22.3 22.2 4.2 15.5 29.7 132.5 9.2 12.7 16.8 9.6 12.6 11.2 12.2 15.2 13.2 30.2 2-Hydroxyisobutyrate 4.9 9.1 9.8 9.5 12.4 19 10.2 26.4 11.2 11.8 28.9 13.9 17.9 16.3 58.2 7.8 6.8 9 4.5 8.6 6.6 5.7 6.5 14.3 13.5 2-Oxoisocaproate 4.8 3 11.1 5.4 17.7 12 8.3 12.2 9 8.6 20.6 6.9 6.8 10.4 15.3 8.1 9.6 15.8 11.1 14.5 11.9 7.8 7.8 12 8.7 3-Hydroxybutyrate 75.3 336.1 215.3 103.1 234.7 283.8 160.3 306 53.2 134.9 127.5 97.5 376.5 202.5 2170.9 311.4 287.6 351.2 323.6 662.9 458.6 333.8 194.6 387.7 611.2 3-Hydroxyisobutyrate/Isobutyrate 19.3 15.2 51.2 14.4 30.9 30.9 30.2 30.9 12.8 19.3 40.9 23.1 13.6 15.7 58.2 13.1 15.2 28.3 20.2 28 24 9.9 20.5 22.1 33.4 Acetate 85.8 21.9 73.5 61.8 313 76.9 153.6 72.7 28.7 73.3 80.6 138.8 53.6 117.5 278.5 114.3 59.2 67.2 67.6 67.8 65.7 47.1 36.7 69.8 95.9 Acetoacetate 5.2 5.2 5.9 3.4 13 14.1 10.1 11.5 3.6 1.6 8 14.7 35.6 16.3 128.1 5.8 6.8 3.2 3.7 9 18.1 13.5 1.4 7.6 6.6 Acetone 2.6 1.6 1.2 2.6 8.4 5.9 1.6 0.5 3.2 2.8 2.2 4.8 6.3 10.1 46.9 2.2 0.6 1.2 2.5 2.3 8.9 6.5 1.4 1 2.6 ADP 59.7 16.5 31.6 60.5 138.3 95.6 66 91.2 41.6 50.2 70.7 120.6 83.3 105 158.1 67.3 71.9 72.6 66.2 72.5 87 87.3 99.6 47.2 93.7 Alanine 131.4 117.4 626.1 198.8 588.7 216.4 498.3 369.1 359.1 466.2 943 232.7 294.4 408.9 186.5 122.6 182.4 327.4 304.1 303.2 165.1 160.8 239.9 263.7 185.7 AMP 29.8 48.5 209.3 18.5 66.3 29.7 40.8 54.5 78.6 226 42.3 91.8 31.5 109.6 64.3 92.8 38.1 40.1 30.5 24.4 86.7 48.3 58.1 186.2 44.9 Arginine 95.5 33.1 111.8 47.1 157.8 151.8 88.9 116.6 127 144 133.6 143.4 103.1 181.2 160.4 55.7 69.6 77.4 100.4 121.2 99 80.3 73.8 82.8 135.9 Asparagine 60.4 18.2 61.9 23.1 49.8 27.3 56.7 73 57.7 70.9 77.5 80.8 39.3 27 58 47.2 32.4 11.5 34.2 45.2 57.2 24.9 32.6 44.3 35.9 Aspartate 42.7 18 62.9 14.2 51.7 33.3 25.2 69.8 47.9 54.1 55.6 104.5 54.9 99.2 52.9 36.5 30.2 15.7 28.5 22.4 46.4 35.8 54.9 52.1 22.8 ATP 163 80.1 74.5 159.4 440.9 294.3 160.2 299.5 175 73.8 205.1 183.3 388.9 226.9 469.4 205 295 247.8 242.8 183.3 183.8 197.1 195.2 195.4 279.5 Betaine/TMAO 8.3 9.3 11.2 8.5 15.4 15.8 17.1 41 74.4 92.2 62.4 123.2 108.1 177.5 240.7 23.3 24 27.6 20.6 23.7 90.4 71.6 66.1 81.7 58.3 Carnitine 10.2 3.6 28.3 13.1 33.5 11.9 13.8 20.4 23 15.2 33.2 10.3 10.3 29.3 29.1 14.8 19.1 20.1 20.6 14.4 22.9 29.5 28.5 13.2 17.4 Choline 17 16.9 67.4 13 43.6 22.2 25.3 33.5 29.3 53.3 43.8 31.3 29.3 45.7 43.4 28.5 23.8 23.8 27.5 23.5 33.3 27.2 25.9 41.2 24.9 Citrate 29.5 18.4 39.2 29 93.8 39.9 44.5 55.6 15.4 24.4 38.9 78.4 85 94.1 87.8 27.4 49.3 33.6 54.2 44 36.7 47.7 55.4 34.7 89.3 Creatine 29.3 56.5 111.7 36.8 74 68.4 67.2 108.2 56.2 68.5 190.7 84.9 174.5 112.6 178.7 67.1 73.2 93.5 85.5 72.6 70.8 63.4 92.2 85.8 99.8 Creatine phosphate 8.2 7.7 12.8 7.1 11.5 9.9 5.8 16.1 4.6 9.4 27.6 7.9 7.8 2.4 15 10.6 9.5 12.9 9.2 9.2 5.7 11.2 4.4 6.3 15.8 Dimethylamine 4.2 0.6 3.7 1.5 2.6 2.6 2.6 3.3 2.9 4.7 5.2 4.7 5.5 4.4 4 2.2 5.7 2.8 1.9 4 3.2 2.2 3.1 2.7 2.8 Dimethyl sulfone 2.6 2.1 8.4 1.3 9.1 4.2 6.8 9.7 3.7 9.6 12.1 5.2 7.8 13.6 10.6 4.1 6.7 9 8.3 6.6 3.4 5.1 7.5 8.4 8.3 Formate 12.3 11.2 90.8 13.1 32 17.9 25.4 34.4 21.5 45.9 33.5 28.2 24.5 26.4 32.7 25.9 23.2 14.4 21.5 20.1 20.2 12.9 30 13.9 17.3 Glucose 3307.5 1909.5 4706.5 2557.7 5555.4 2970.4 3982.7 5273 3186.9 5059.9 4936 4661.7 3500.5 3972.4 3550.1 2466.3 4592.9 5112.8 5007.5 4791.9 2834 3736.1 2598.1 4122.9 2735.3 Glutamate 84.9 30.2 161.7 71.7 228 77.6 144.5 130.9 138.7 216 110.5 192.5 74.4 145.6 100.2 77.2 83.9 78.1 71.9 89.7 124 120.9 71.7 108.2 92.8 Glutamine 298.9 177.1 92.1 165.3 461.1 278.4 284.2 561.6 428.4 453.5 608.9 431.4 428.2 562.9 552.4 225.1 300.3 255.2 284.5 277.2 279.9 256.2 266.5 309.9 430.9 Glutathione 343.8 133.2 311.2 227 633.4 423.8 395.4 539.6 312.3 381.9 322.7 426.9 377 575.6 435.8 373.1 429.6 452.3 417.5 425.3 401.3 417.9 339.2 413.5 389.5 Glycerol 37.6 26.2 71.3 30.9 80.8 21.7 66.6 104 95.3 89.7 97.1 84.7 80.7 106.9 66.9 146.2 92.9 77 120.2 109.4 66.4 29.8 39.7 94.3 109.4 Glycine 146.3 104.6 309.8 102.4 282 147.2 157.3 285.1 101.8 259.1 205.1 232 211.4 262 154.4 120.9 135 167.7 189.8 151.1 218 173.3 190.7 221.7 253.2 Histidine 33.6 51.3 61.3 18.4 30.1 14 30.9 72.6 52.3 59.1 106.5 39.3 25.5 84.7 33.2 25.4 43.3 41 26.1 26.9 40.1 24.7 31.5 39.9 50.3 Homocysteine 64.9 21.6 108.5 48.8 95.1 45 75.9 72.7 64.3 105.9 97.9 74.3 58.7 28.7 93 64.2 85 99.2 106.8 103.9 93 77.2 113.3 92.5 117.1 Inosine monophosphate (IMP) 19.4 13.7 42.6 9.1 38 16.5 25.2 22.5 20 97.3 13.6 38.7 25.3 56.6 21.2 59.4 22.5 19 15.9 11.9 26.3 16.8 20.6 59.9 6.5 Isoleucine 48 63.3 144 41 116.2 65 74 95.3 51 89.4 102.6 94.6 65.1 91.4 93.8 70.6 70.9 118 68.1 86.7 74.2 69.4 46 68.5 47.6 Lactate 1208.1 1157.4 3333.7 1246.8 3324.5 1827.7 2530.9 1796.1 1790.1 1384 4318.2 1447.5 1546.5 2104.5 2129.3 1181.3 1366.7 2146.6 1589.1 1573.3 844.7 830.8 1664.9 1184.6 1308.1 Leucine 75.5 119.6 162.5 63.6 200 124.8 131.2 157.1 86.3 138 178.7 132.9 93 124.1 109.1 85.9 117.6 170.8 113.4 123.8 112.1 103.4 93 114.2 87.4 Lysine 132.9 90.9 256.4 109.7 235.7 140.3 172.2 211.9 189.1 227 315.6 252.4 169.9 221.4 133.4 103.4 122.4 145 145.3 113.6 80.6 104.3 93.7 94.9 82.8 Malonate 50.3 30.6 49.8 32.3 112.3 66.3 87.9 92.9 56.8 56 76.6 110.3 84.9 132.7 140.9 52.9 64.4 68 53.9 79.5 60.3 46.7 53.3 61 54.5 Methionine 40.5 29.4 61.6 29.2 83.6 20.6 72.3 65.2 56 58.9 148 76 51.1 86.7 34.4 29.2 34.7 48.1 36.8 44.5 41.5 22.4 37.9 44.5 42.4 NAD+ 25.8 16.5 7.5 5.7 5 13.7 15.9 23.3 7.7 12.4 15.8 15.2 18.5 27.1 15 9.4 12.9 16.7 11.6 13.8 3.9 11.6 5 13.4 6.2 Acetylcarnitine 10.1 11.7 21.9 8.4 20 13.9 15.9 21.5 13.5 21.5 21.3 15.5 19.1 23.6 19.9 17.5 18.2 18.2 17.6 17.2 15 15.3 19.8 14.3 16 Ornithine 29 18.9 46.6 28.7 50.9 26.8 38 62.6 15.1 37.9 58.9 55.6 39.5 62.9 26 15.2 26.4 32.8 29.7 28.8 20.5 25.9 20.8 26.2 26.5 Phenylalanine 35.6 32.3 54.5 30.9 58.9 44.6 54.7 92 38.9 55.7 99.5 54.9 68.5 50.4 48.6 47.9 56 52.1 45.8 36.3 35.1 31.1 37.2 32 46 Proline 74.5 44.9 104.5 70.6 86.6 126.1 119.5 129.4 102.6 151.9 315.3 129.3 139.2 213.7 115.3 50.5 95.6 80 90.6 130.7 75 65.2 61.3 81.5 143.9 Pyruvate 7.4 1.4 15.9 5.2 13 2.2 12.9 7.4 5.2 10.6 9.4 15.2 7.3 19.1 8.4 5 5.6 6.5 3.9 5.4 12.1 6.7 4.3 10.8 6 Sarcosine 7 5.9 4.2 2.1 6.8 2.7 6.2 11.4 3.7 2.1 6.7 10 4.3 6.7 3.9 3.5 4 5.4 3.1 3 5.9 1.6 4.3 3.2 2.7 Serine 86 40.4 152.3 20.2 66.1 72.1 117.7 154.4 147.2 156.5 196 79 24.9 161.4 60.7 143.8 80.2 87.6 98.6 98.8 71.4 58.1 139.1 114.2 109 Succinate 5.7 3.2 8.6 5.9 19.7 5 13.3 4.3 10.8 3.1 7.5 11.1 11 11.3 21.6 4.9 8.4 6.4 11.7 11.6 5.1 8.9 6 6.4 5.1 Taurine 237 317.7 767.8 289.3 927.3 408.2 490.6 513.3 397.7 736.6 914.8 462.6 885.6 993.7 1207.9 466.3 487.8 462.8 630.8 463 612.8 503.6 752.7 722.9 917.4 Threonine 69.2 61.1 177.4 36.2 180.2 74.5 68.9 104.7 104.4 152 120.2 78.4 35.8 104.9 94.6 77.2 79 86.9 77.9 80.4 73.6 41.2 43.6 47.8 84.1 Trimethylamine N-oxide (TMAO)/Betaine 18.7 9.8 41 21.6 35.9 27 40.2 51 17.6 28.7 28.8 28.2 15 28.7 29.2 32.7 32.7 41.2 41.6 41.6 30 25.7 54.9 26.8 53.8 Tryptophan 35.4 9.9 55.5 21 43.6 32.1 45.3 59.6 35.5 53.7 74.9 63.3 51.5 77.1 47.8 11.1 41.4 32.6 35.8 35 46.9 23.5 32.2 43.7 30.5 Tyrosine 47 26 67.4 41.3 68.8 41.5 57.9 66.9 52.6 67.7 117.1 54.2 43.9 46 31.8 35.1 49.1 60.9 63.4 45.8 45.2 29.6 41.7 65 53.1 Valine 119.8 161.7 265.5 85.4 233.9 134.2 164.4 170.2 104.5 169.2 225.4 216 122.3 176.7 141.5 94.6 118.2 189.1 121.8 147.7 123.8 117.9 89.4 112.5 83.3 NMR_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name KEGG ID 2-Hydroxybutyrate C05984 2-Hydroxyisobutyrate 2-Oxoisocaproate C00233 3-Hydroxybutyrate C01089 3-Hydroxyisobutyrate/Isobutyrate C06001/C02632 Acetate C00033 Acetoacetate C00164 Acetone C00207 ADP C00008 Alanine C00041 AMP C00020 Arginine C00062 Asparagine C00152 Aspartate C00049 ATP C00002 Betaine/TMAO C00719/C01104 Carnitine C00318 Choline C00114 Citrate C00158 Creatine C00300 Creatine phosphate C02305 Dimethylamine C00543 Dimethyl sulfone C11142 Formate C00058 Glucose C00031 Glutamate C00217 Glutamine C00064 Glutathione C00051 Glycerol C00116 Glycine C00037 Histidine C00135 Homocysteine C00155 Inosine monophosphate (IMP) C00130 Isoleucine C00407 Lactate C00186 Leucine C00123 Lysine C00047 Malonate C00383 Methionine C00073 NAD+ C00003 Acetylcarnitine C02571 Ornithine C00077 Phenylalanine C00079 Proline C00148 Pyruvate C00022 Sarcosine C00213 Serine C00065 Succinate C00042 Taurine C00245 Threonine C00188 Trimethylamine N-oxide (TMAO)/Betaine C01104/C00719 Tryptophan C00078 Tyrosine C00082 Valine C00183 METABOLITES_END #END