#METABOLOMICS WORKBENCH hormel101_20170623_140813 DATATRACK_ID:1094 STUDY_ID:ST000645 ANALYSIS_ID:AN000977 PROJECT_ID:PR000460 VERSION 1 CREATED_ON June 25, 2017, 10:25 pm #PROJECT PR:PROJECT_TITLE The dystrophic muscle metabolome: effects of exercise and NO donor therapy PR:PROJECT_SUMMARY "In Duchenne and Becker muscular dystrophy (DMD, BMD), loss of the cytoskeletal PR:PROJECT_SUMMARY protein dystrophin weakens the sarcolemma and disrupts cellular signaling, PR:PROJECT_SUMMARY rendering the diseased muscles susceptible to contractioninduced damage. We and PR:PROJECT_SUMMARY others have shown that loss of neuronal nitric oxide synthase (nNOSμ) from the PR:PROJECT_SUMMARY sarcolemma of dystrophin-deficient muscle causes functional muscle ischemia PR:PROJECT_SUMMARY during exercise due to unopposed sympathetic vasoconstriction, thereby PR:PROJECT_SUMMARY exacerbating fatigue and injury of the diseased muscles. Genetic and PR:PROJECT_SUMMARY pharmacologic strategies targeting nNOSμ-NO signaling ameliorate functional PR:PROJECT_SUMMARY muscle ischemia, as well as many other features of the dystrophic phenotype in PR:PROJECT_SUMMARY the mdx mouse model of DMD/BMD. These findings suggest that the therapeutic PR:PROJECT_SUMMARY benefit of NO likely extends beyond its vascular effects. A growing body of PR:PROJECT_SUMMARY evidence indicates that NO directly influences muscle metabolism through effects PR:PROJECT_SUMMARY on glucose transport as well as mitochondrial biogenesis and function. Both PR:PROJECT_SUMMARY nNOS-/- mice and mdx mice exhibit muscle mitochondrial dysfunction, decreased PR:PROJECT_SUMMARY resistance to fatigue, and exercise-induced muscle injury, suggesting a causal PR:PROJECT_SUMMARY role of nNOSμ-NO deficiency. However, the specific metabolic changes resulting PR:PROJECT_SUMMARY from reduced NO signaling that might render dystrophic muscle susceptible to PR:PROJECT_SUMMARY fatigue and use-dependent injury remain poorly defined. Therefore, the goal of PR:PROJECT_SUMMARY this pilot metabolomics study is to identify the unique biochemical profiles of PR:PROJECT_SUMMARY skeletal and cardiac muscles of mdx mice to gain further mechanistic insight PR:PROJECT_SUMMARY into the pathophysiological role of NO deficiency in muscular dystrophy. In Aim PR:PROJECT_SUMMARY 1, we will characterize the skeletal and cardiac muscle metabolomes of mdx and PR:PROJECT_SUMMARY nNOS-/- mice at rest and following a single bout of treadmill exercise with the PR:PROJECT_SUMMARY goal of discovering common metabolic signatures caused by loss of NO signaling. PR:PROJECT_SUMMARY In Aim 2, we will evaluate the potential of a NO donor drug that is under PR:PROJECT_SUMMARY development as a therapeutic for DMD/BMD to improve the skeletal and cardiac PR:PROJECT_SUMMARY muscle metabolomes in mdx mice. As a result of this pilot study, we hope to gain PR:PROJECT_SUMMARY new understanding of the metabolic derangements in dystrophin-deficient muscle, PR:PROJECT_SUMMARY insight into the therapeutic effects of NO replacement, and to identify new PR:PROJECT_SUMMARY pathogenic mechanisms and putative therapeutic targets that will form the basis PR:PROJECT_SUMMARY of future grant applications." PR:INSTITUTE Mayo Clinic PR:LAST_NAME Thomas PR:FIRST_NAME Gail PR:ADDRESS Penn State Hershey Heart and Vascular Institute Penn State College of Medicine PR:ADDRESS 500 University Drive, MC H047 Hershey, PA 17033 PR:EMAIL gthomas4@hmc.psu.edu PR:PHONE 717-531-0003, ext. 287087 #STUDY ST:STUDY_TITLE Effects of Exercise on Dystrophic Mouse Muscle Amino Acids (part II) ST:STUDY_SUMMARY We will use male C57BL10, mdx, and nNOS-/- mice (n = 10 each group) to ST:STUDY_SUMMARY characterize the skeletal and cardiac muscle metabolomes. Half of the mice in ST:STUDY_SUMMARY each group will remain sedentary while the other half will be subjected to a ST:STUDY_SUMMARY single bout of treadmill exercise to exhaustion. Mice will be euthanized ST:STUDY_SUMMARY immediately postexercise and blood, hearts, and hindlimb muscles will be ST:STUDY_SUMMARY harvested and frozen as detailed in the General Methods. The heart and ST:STUDY_SUMMARY gastrocnemius, soleus, and quadriceps muscles of one hindlimb will be sent to ST:STUDY_SUMMARY the Mayo Clinic Metabolomics Resource Core. The heart and quadriceps muscle will ST:STUDY_SUMMARY be used for untargeted metabolomics profiling (LC/MS) while the gastrocnemius ST:STUDY_SUMMARY and soleus muscles will be used for targeted analyses of amino acids plus amino ST:STUDY_SUMMARY metabolites, non-esterified fatty acids, and citric acid cycle intermediates. ST:INSTITUTE Mayo Clinic ST:LAST_NAME Thomas ST:FIRST_NAME Gail ST:ADDRESS Penn State Hershey Heart and Vascular Institute Penn State College of Medicine ST:ADDRESS 500 University Drive, MC H047 Hershey, PA 17033 ST:EMAIL gthomas4@hmc.psu.edu ST:PHONE 717-531-0003, ext. 287087 #SUBJECT SU:SUBJECT_TYPE Mouse SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - ms5942-1 Time point:Sedentary | Grouping:SED-BL10 Control Date of study=10/15/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-2 Time point:Sedentary | Grouping:SED-BL10 Control Date of study=10/21/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-3 Time point:Sedentary | Grouping:SED-BL10 Control Date of study=10/28/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-4 Time point:Sedentary | Grouping:SED-BL10 Control Date of study=11/23/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-5 Time point:Sedentary | Grouping:SED-BL10 Control Date of study=11/25/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-6 Time point:Post Run | Grouping:RUN-BL10 Control Date of study=10/20/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-7 Time point:Post Run | Grouping:RUN-BL10 Control Date of study=10/21/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-8 Time point:Post Run | Grouping:RUN-BL10 Control Date of study=10/28/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-9 Time point:Post Run | Grouping:RUN-BL10 Control Date of study=10/30/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-10 Time point:Post Run | Grouping:RUN-BL10 Control Date of study=11/25/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-11 Time point:Sedentary | Grouping:SED-nNOS-/- Date of study=10/20/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-12 Time point:Sedentary | Grouping:SED-nNOS-/- Date of study=10/21/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-13 Time point:Sedentary | Grouping:SED-nNOS-/- Date of study=10/28/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-14 Time point:Sedentary | Grouping:SED-nNOS-/- Date of study=11/23/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-15 Time point:Sedentary | Grouping:SED-nNOS-/- Date of study=11/25/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-16 Time point:Post Run | Grouping:RUN-nNOS-/- Date of study=10/20/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-17 Time point:Post Run | Grouping:RUN-nNOS-/- Date of study=10/21/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-18 Time point:Post Run | Grouping:RUN-nNOS-/- Date of study=10/28/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-19 Time point:Post Run | Grouping:RUN-nNOS-/- Date of study=10/30/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-20 Time point:Post Run | Grouping:RUN-nNOS-/- Date of study=11/25/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-21 Time point:Sedentary | Grouping:SED-MDX Date of study=10/20/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-22 Time point:Sedentary | Grouping:SED-MDX Date of study=10/21/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-23 Time point:Sedentary | Grouping:SED-MDX Date of study=10/28/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-24 Time point:Sedentary | Grouping:SED-MDX Date of study=11/23/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-25 Time point:Sedentary | Grouping:SED-MDX Date of study=11/25/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-26 Time point:Post Run | Grouping:RUN-MDX Date of study=10/20/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-27 Time point:Post Run | Grouping:RUN-MDX Date of study=10/21/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-28 Time point:Post Run | Grouping:RUN-MDX Date of study=10/28/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-29 Time point:Post Run | Grouping:RUN-MDX Date of study=10/30/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse SUBJECT_SAMPLE_FACTORS - ms5942-30 Time point:Post Run | Grouping:RUN-MDX Date of study=11/25/2015; Gender=Male; Sample type=Tissue-Gastrocnemius; Species=Mouse #COLLECTION CO:COLLECTION_SUMMARY Blood and Tissue Harvesting: Mice will be euthanized and blood will be taken by CO:COLLECTION_SUMMARY cardiac puncture, centrifuged, and the plasma will be stored at -80ºC. The CO:COLLECTION_SUMMARY heart, diaphragm, and muscles of both hindlimbs (gastrocnemius, soleus, CO:COLLECTION_SUMMARY quadriceps) will be dissected and snap frozen in liquid nitrogen. Tibialis CO:COLLECTION_SUMMARY anterior and plantaris muscles from both hindlimbs also will be dissected and CO:COLLECTION_SUMMARY one set will be snap frozen in liquid nitrogen while the other set will be CO:COLLECTION_SUMMARY mounted in OCT and frozen in isopentane cooled by liquid nitrogen. Cryosections CO:COLLECTION_SUMMARY will be used to evaluate disease activity and exercise-induced muscle injury by CO:COLLECTION_SUMMARY staining with: (a) hematoxylin and eosin to assess gross morphology, cellular CO:COLLECTION_SUMMARY infiltration, and necrosis, (b) anti-F4/80 to label macrophages, and (c) CO:COLLECTION_SUMMARY anti-IgG or IgM to label damaged muscle fibers. #TREATMENT TR:TREATMENT_SUMMARY "Studies will be performed in 12-16 week old male mdx mice, C57BL10 control TR:TREATMENT_SUMMARY mice, and nNOS-/- mice obtained from Jackson Laboratory. All protocols will be TR:TREATMENT_SUMMARY approved by the Penn State College of Medicine Institutional Animal Care and Use TR:TREATMENT_SUMMARY Committee. Treadmill Exercise: Mice will run on a horizontal treadmill (Columbus TR:TREATMENT_SUMMARY Instruments) to assess fatigue by a single bout of exercise beginning at 5 m/min TR:TREATMENT_SUMMARY for 5 min followed by 1 m/min increases every minute until exhaustion. Electric TR:TREATMENT_SUMMARY shocks will not be used to stimulate running due to adverse effects in mdx TR:TREATMENT_SUMMARY mice." #SAMPLEPREP SP:SAMPLEPREP_SUMMARY mouse gastrocnemius amino acids #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Waters Acquity CH:COLUMN_NAME Waters Acquity BEH C18 (150 x 2.1mm, 1.7um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME Thermo Quantum Ultra MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE ESI MS:ION_MODE POSITIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS picomol/mg MS_METABOLITE_DATA_START Samples ms5942-1 ms5942-2 ms5942-3 ms5942-4 ms5942-5 ms5942-6 ms5942-7 ms5942-8 ms5942-9 ms5942-10 ms5942-11 ms5942-12 ms5942-13 ms5942-14 ms5942-15 ms5942-16 ms5942-17 ms5942-18 ms5942-19 ms5942-20 ms5942-21 ms5942-22 ms5942-23 ms5942-24 ms5942-25 ms5942-26 ms5942-27 ms5942-28 ms5942-29 ms5942-30 Factors Time point:Sedentary | Grouping:SED-BL10 Control Time point:Sedentary | Grouping:SED-BL10 Control Time point:Sedentary | Grouping:SED-BL10 Control Time point:Sedentary | Grouping:SED-BL10 Control Time point:Sedentary | Grouping:SED-BL10 Control Time point:Post Run | Grouping:RUN-BL10 Control Time point:Post Run | Grouping:RUN-BL10 Control Time point:Post Run | Grouping:RUN-BL10 Control Time point:Post Run | Grouping:RUN-BL10 Control Time point:Post Run | Grouping:RUN-BL10 Control Time point:Sedentary | Grouping:SED-nNOS-/- Time point:Sedentary | Grouping:SED-nNOS-/- Time point:Sedentary | Grouping:SED-nNOS-/- Time point:Sedentary | Grouping:SED-nNOS-/- Time point:Sedentary | Grouping:SED-nNOS-/- Time point:Post Run | Grouping:RUN-nNOS-/- Time point:Post Run | Grouping:RUN-nNOS-/- Time point:Post Run | Grouping:RUN-nNOS-/- Time point:Post Run | Grouping:RUN-nNOS-/- Time point:Post Run | Grouping:RUN-nNOS-/- Time point:Sedentary | Grouping:SED-MDX Time point:Sedentary | Grouping:SED-MDX Time point:Sedentary | Grouping:SED-MDX Time point:Sedentary | Grouping:SED-MDX Time point:Sedentary | Grouping:SED-MDX Time point:Post Run | Grouping:RUN-MDX Time point:Post Run | Grouping:RUN-MDX Time point:Post Run | Grouping:RUN-MDX Time point:Post Run | Grouping:RUN-MDX Time point:Post Run | Grouping:RUN-MDX Histidine 67.4 111.8 147.0 96.1 97.6 124.1 104.0 82.2 156.0 82.6 89.1 145.5 109.4 95.6 90.2 72.8 120.4 146.8 157.0 138.3 167.0 132.4 132.9 121.6 198.3 123.2 158.2 116.3 181.1 131.7 Hydroxyproline 17.7 285.9 331.8 141.7 138.4 262.2 244.7 158.1 224.2 188.6 156.4 164.0 268.2 134.7 82.6 114.4 178.8 254.2 305.5 299.3 221.9 267.7 264.6 287.0 237.1 119.9 140.0 218.4 209.7 227.3 1-Methylhistidine 1.5 5.5 2.3 1.9 2.2 3.3 3.4 2.1 1.2 1.2 7.7 4.3 2.0 4.5 2.8 3.7 7.7 5.0 4.7 4.8 2.4 2.7 2.8 5.3 17.0 1.7 7.8 5.7 4.7 4.0 3-Methylhistidine 2.0 2.2 7.8 2.1 2.7 4.2 2.3 1.3 1.4 2.2 3.5 4.0 6.1 3.1 2.4 4.6 3.6 3.5 2.6 0.6 5.1 6.0 3.4 5.6 5.6 4.6 4.0 5.5 8.4 5.0 Asparagine 42.1 54.8 64.0 55.6 49.0 62.8 68.5 43.0 75.1 53.6 57.0 49.2 64.6 50.6 64.0 33.7 60.0 62.5 61.8 66.4 87.7 49.8 73.5 68.8 105.8 72.5 60.0 53.1 74.6 78.1 Phosphoethanolamine 3.1 5.2 5.3 5.7 5.5 4.8 4.9 5.1 6.8 5.2 5.5 5.9 7.0 6.0 3.7 6.0 5.4 6.7 7.0 8.9 38.7 16.9 15.8 15.4 33.5 20.3 16.5 18.7 16.8 31.5 Arginine 70.9 157.3 140.5 98.4 74.6 124.2 95.0 51.5 126.4 70.2 98.9 150.6 207.7 121.3 135.0 51.7 93.1 128.5 105.9 150.3 128.5 177.1 143.2 195.6 189.2 82.3 138.2 100.5 135.9 164.4 Carnosine 1142.1 1909.2 1618.5 2047.3 1598.4 2091.6 2324.7 1898.4 2008.3 1739.0 1458.5 1571.8 1977.1 2003.0 1490.7 1237.7 1986.2 2259.6 1998.6 2761.7 1331.8 1152.7 1364.2 1716.6 1765.2 935.9 1316.3 1304.3 1428.1 1014.6 Taurine 9344.2 11962.3 15132.5 13453.3 12309.9 13125.4 14943.6 11927.8 14208.1 11954.2 12507.8 10931.2 12443.4 14011.8 9892.2 6562.8 11375.3 11363.3 9728.9 15922.4 15846.4 11088.0 13306.8 16013.6 21328.6 10856.7 13729.8 10820.9 14928.4 12887.9 Anserine 1945.5 2352.9 2379.1 3283.0 2740.1 2890.1 2974.2 2668.3 2583.4 2580.9 2813.0 2170.0 2984.1 3517.0 2323.4 1301.2 2669.1 2588.4 2116.6 4298.3 1297.7 1004.5 1362.2 1459.4 2284.5 882.8 1404.6 1293.2 1653.2 798.2 Serine 105.0 248.5 274.6 190.1 186.0 209.5 232.1 132.5 235.2 168.3 232.2 192.3 226.4 189.4 178.0 112.6 204.4 232.9 230.2 281.1 261.4 159.0 231.2 183.8 296.9 155.5 190.2 161.6 206.3 197.0 Glutamine 552.3 1096.6 1187.0 992.2 1072.7 829.9 802.6 699.5 895.1 777.4 1093.2 1001.0 979.0 1164.5 624.1 451.0 919.7 827.4 932.8 889.1 1276.7 1047.2 1297.3 1051.4 1772.3 893.5 1057.0 839.1 1185.7 1347.9 Ethanolamine 16.8 18.3 41.3 16.4 18.3 20.1 35.0 19.4 24.9 16.6 16.8 21.3 17.9 18.5 11.7 10.1 20.1 22.8 17.5 21.8 31.6 16.9 24.0 17.2 32.6 19.6 21.8 17.1 25.2 27.1 Glycine 517.2 1888.2 2010.9 1549.3 1347.2 1925.9 1780.5 1426.0 1862.4 1459.9 1911.4 1294.1 1944.5 1855.3 1266.2 999.0 1715.2 1928.0 1908.3 1768.7 1867.7 1199.0 1952.5 1587.4 1982.7 1199.2 1241.2 1902.1 1742.8 2123.6 Aspartic Acid 52.7 38.4 52.0 59.6 40.4 95.2 93.1 40.7 96.9 55.2 77.1 68.9 68.0 95.0 48.8 40.9 93.3 79.2 90.7 78.1 162.4 92.5 103.0 96.6 165.2 90.0 102.9 52.8 121.1 129.5 Sarcosine -0.2 7.9 4.3 5.0 4.1 5.0 4.9 3.6 6.1 4.0 2.6 2.5 2.6 4.7 4.7 0.9 4.3 2.9 3.6 5.2 2.7 2.8 2.5 3.2 3.4 1.5 3.0 3.2 3.0 2.5 Citrulline 36.6 108.4 122.8 112.1 92.3 107.0 135.7 100.7 117.6 101.0 162.9 149.4 177.6 151.2 126.6 79.4 156.5 132.5 152.4 170.1 85.6 98.5 95.6 105.5 116.0 75.4 101.2 58.8 105.7 92.9 Glutamic Acid 248.6 215.5 323.2 456.8 301.1 404.9 406.7 488.6 553.2 495.0 469.1 390.1 372.5 490.3 159.8 278.9 431.0 481.9 417.4 501.9 774.8 474.9 549.2 553.0 904.9 514.3 534.0 377.9 685.6 739.0 beta-Alanine 55.0 71.7 89.2 45.3 34.8 68.0 75.1 51.2 61.3 48.5 30.8 36.0 71.2 50.0 24.8 31.5 42.4 74.4 68.7 89.4 48.9 53.5 44.3 58.9 51.4 27.3 39.9 32.5 43.9 37.5 Threonine 85.8 159.7 161.7 161.3 133.8 161.4 170.7 114.9 170.4 126.5 138.5 131.5 118.3 155.2 134.0 77.1 142.7 153.1 133.5 184.5 188.8 149.2 164.8 177.3 229.5 151.3 168.6 137.3 173.0 185.9 Alanine 724.8 1450.6 1496.8 1366.4 1292.7 1360.1 1366.0 935.3 1378.9 1121.3 1315.5 1138.6 1447.0 1462.3 830.4 738.7 1343.3 1269.5 1276.8 1640.2 1470.9 1285.6 1314.9 1356.0 1770.0 1072.9 1362.2 1036.8 1491.7 1390.6 gamma-Amino-N-butyric-acid 0.26 0.30 0.90 0.65 0.91 0.80 0.70 0.86 1.08 0.84 0.75 0.50 1.97 1.01 1.07 0.93 1.26 1.27 0.88 1.57 1.46 0.79 0.93 0.84 1.40 0.74 0.84 0.64 0.92 1.22 alpha-Aminoadipic-acid 0.4 1.0 0.5 1.5 1.1 0.5 0.5 1.3 1.9 0.7 0.5 0.9 0.5 0.5 0.1 0.1 0.7 0.7 0.3 2.4 4.4 1.9 2.1 2.1 2.8 2.1 1.2 1.8 2.0 6.2 beta-Aminoisobutyric-acid 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Proline 96.9 164.2 176.2 102.3 82.9 145.2 190.3 108.2 176.3 103.3 97.3 111.4 126.5 103.8 122.1 74.6 140.7 137.3 140.1 200.7 137.0 115.1 114.2 145.4 143.9 111.4 119.3 99.9 132.9 105.7 Hydroxylysine 1 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Hydroxylysine 2 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.2 0.1 0.1 0.1 0.0 0.1 0.1 0.2 0.1 0.2 alpha-Amino-N-butyric-acid 2.2 4.1 3.5 2.2 3.0 2.4 2.3 1.7 3.2 2.1 1.5 1.3 3.5 2.4 2.1 1.0 1.1 2.9 2.2 2.9 1.8 2.4 1.9 2.1 2.5 1.9 1.6 1.4 1.8 2.7 Ornithine 4.6 23.2 28.4 18.2 18.2 22.0 26.6 10.7 26.5 15.0 16.6 22.4 20.7 14.7 25.8 9.4 17.7 25.7 16.1 38.9 39.3 28.8 28.8 39.1 48.6 20.2 31.5 19.9 25.7 24.3 Cystathionine 1 1.4 4.6 6.9 2.8 2.4 3.6 6.3 3.0 6.6 2.3 3.4 4.3 4.5 2.6 1.6 1.7 3.4 3.9 2.5 7.4 6.4 3.3 4.0 4.9 7.2 2.9 2.7 1.8 4.6 3.9 Cystathionine 2 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Lysine 186.7 462.7 420.0 257.9 216.0 311.5 241.0 134.7 382.0 188.8 256.0 415.1 621.7 330.6 343.8 134.0 227.9 351.8 220.4 337.9 371.9 467.6 434.5 450.0 532.3 261.7 356.8 309.4 445.9 529.4 Cystine 7.3 7.8 11.4 9.7 9.1 8.9 7.8 10.3 11.8 8.4 7.5 9.0 8.8 8.4 13.1 6.5 7.4 8.9 10.2 11.9 27.5 14.7 15.1 16.2 20.7 22.2 15.3 15.0 15.2 24.2 Tyrosine 27.8 40.9 54.2 53.6 43.2 70.7 75.7 56.5 76.5 54.3 41.8 48.5 47.7 52.0 69.2 44.2 80.6 80.3 70.2 100.9 79.1 49.6 55.0 87.2 92.9 75.0 66.1 63.6 80.9 76.0 Methionine 34.7 35.9 42.0 47.4 35.6 46.7 52.2 31.0 44.2 31.6 35.9 41.7 36.2 40.7 44.7 27.1 50.5 52.3 45.0 61.1 68.0 48.9 58.1 74.6 78.2 50.6 61.5 44.4 55.5 47.2 Valine 70.8 98.9 125.9 102.2 92.5 117.0 125.8 101.6 145.3 90.7 85.4 83.9 100.2 88.6 96.1 62.2 96.9 103.2 100.0 142.3 152.1 93.8 110.1 116.2 161.8 106.9 108.7 77.9 123.3 111.4 Isoleucine 36.6 33.5 47.9 35.2 37.8 48.7 47.5 38.4 55.8 32.2 31.1 35.1 39.6 36.7 39.1 24.7 38.1 38.9 41.1 54.1 57.4 42.6 45.6 46.7 59.4 43.7 44.6 33.2 50.9 41.5 allo-Isoleucine 0.0 0.1 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.1 0.0 0.0 0.0 0.0 0.0 0.0 0.1 0.1 0.0 0.0 0.1 0.1 0.0 0.0 0.0 0.0 0.0 Homocystine 0.4 0.9 0.8 1.1 0.9 1.5 1.8 0.9 1.5 1.1 1.0 1.1 1.5 1.5 0.9 0.5 1.7 2.0 1.4 2.3 0.9 1.0 0.9 1.1 1.7 0.5 0.7 0.8 1.0 0.9 Leucine 65.0 62.3 82.1 63.4 66.6 75.6 84.7 62.0 91.8 51.3 63.4 75.2 73.9 64.1 64.7 41.8 67.3 67.6 69.5 100.2 145.2 97.5 108.5 116.2 159.0 96.4 97.8 67.1 101.2 94.7 Phenylalanine 33.8 38.1 45.6 41.0 39.5 51.9 56.7 44.9 59.2 40.6 36.0 45.1 42.1 38.3 40.1 31.3 52.5 56.8 49.8 77.1 77.5 47.4 54.1 63.6 81.9 59.5 64.2 48.0 62.6 62.5 Tryptophan 8.0 8.0 10.2 11.5 10.1 21.1 22.7 17.7 24.7 16.2 10.4 10.5 9.5 11.7 15.2 11.4 22.8 21.4 15.9 25.7 15.4 11.3 11.8 17.8 20.6 18.8 16.8 15.6 17.2 16.5 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Histidine Hydroxyproline 1-Methylhistidine 3-Methylhistidine Asparagine Phosphoethanolamine Arginine Carnosine Taurine Anserine Serine Glutamine Ethanolamine Glycine Aspartic Acid Sarcosine Citrulline Glutamic Acid beta-Alanine Threonine Alanine gamma-Amino-N-butyric-acid alpha-Aminoadipic-acid beta-Aminoisobutyric-acid Proline Hydroxylysine 1 Hydroxylysine 2 alpha-Amino-N-butyric-acid Ornithine Cystathionine 1 Cystathionine 2 Lysine Cystine Tyrosine Methionine Valine Isoleucine allo-Isoleucine Homocystine Leucine Phenylalanine Tryptophan METABOLITES_END #END