#METABOLOMICS WORKBENCH sdasari_20140930_9217761_mwtab.txt DATATRACK_ID:163 STUDY_ID:ST000115 ANALYSIS_ID:AN000195 PROJECT_ID:PR000104 VERSION 1 CREATED_ON 2016-09-17 #PROJECT PR:PROJECT_TITLE Impact of insulin deprivation and treatment on sphingolipid distribution in PR:PROJECT_TITLE muscle subcellular compartments of streptozotocin-diabetic C57Bl/6 mice. PR:PROJECT_TYPE Targeted metabolomics PR:PROJECT_SUMMARY Insulin deprivation in type 1 diabetes (T1D) individuals increases lipolysis PR:PROJECT_SUMMARY plasma free fatty acids (FFA) concentration, which can stimulate synthesis of PR:PROJECT_SUMMARY bioactive lipids such as ceramides (Cer) and long-chain fatty acid-CoAs PR:PROJECT_SUMMARY Ceramide was shown to decrease muscle insulin sensitivity, and at mitochondrial PR:PROJECT_SUMMARY it stimulates reactive oxygen species production. Here, we show that insulin PR:PROJECT_SUMMARY in streptozotocin diabetic C57BL/6 mice increases quadriceps muscle Cer PR:PROJECT_SUMMARY which was correlated with a concomitant decrease in the body fat and increased PR:PROJECT_SUMMARY FFA, glycosylated hemoglobin level (%Hb A1c), and muscular LCFa-CoA content. PR:PROJECT_SUMMARY alternations were accompanied by an increase in protein expression in LCFa-CoA PR:PROJECT_SUMMARY Cer synthesis (FATP1/ACSVL5, CerS1, CerS5), a decrease in the expression of PR:PROJECT_SUMMARY implicated in muscle insulin sensitivity (GLUT4, GYS1), and inhibition of PR:PROJECT_SUMMARY signaling cascade by Akt? and GYS3? phosphorylation under acute insulin PR:PROJECT_SUMMARY Both the content and composition of sarcoplasmic fraction sphingolipids were PR:PROJECT_SUMMARY affected by insulin deprivation, whereas mitochondrial fraction sphingolipids PR:PROJECT_SUMMARY stable. The observed effects of insulin deprivation were reversed, except for PR:PROJECT_SUMMARY and composition of LCFa-CoA, CerS protein expression, GYS1 gene expression, and PR:PROJECT_SUMMARY status of Akt and GYS3? when exogenous insulin was provided by subcutaneous PR:PROJECT_SUMMARY implants. Principal component analysis and Pearson's correlation analysis PR:PROJECT_SUMMARY close relationships between the features of the diabetic phenotype, the content PR:PROJECT_SUMMARY LCFa-CoAs and Cers containing C18-fatty acids in sarcoplasm, but not in PR:PROJECT_SUMMARY Insulin replacement did not completely rescue the phenotype, especially PR:PROJECT_SUMMARY the content of LCFa-CoA, or proteins implicated in Cer synthesis and muscle PR:PROJECT_SUMMARY sensitivity. These persistent changes might contribute to muscle insulin PR:PROJECT_SUMMARY observed in T1D individuals. PR:INSTITUTE Mayo Clinic PR:DEPARTMENT Endocrinology PR:LABORATORY Dr. Sreekumaran Nair's lab PR:LAST_NAME Nair PR:FIRST_NAME Sreekumaran PR:ADDRESS - PR:EMAIL Dasari.Surendra@mayo.edu PR:PHONE - PR:FUNDING_SOURCE R01-DK-41973, UL1 TR000135, the David Murdock Dole Professorship (K. S. Nair), PR:FUNDING_SOURCE the Stephenson Fellowship (P. Zabielski). PR:PROJECT_COMMENTS 24368672 #STUDY ST:STUDY_TITLE Impact of insulin deprivation and treatment on sphingolipid distribution in ST:STUDY_TITLE muscle subcellular compartments of streptozotocin-diabetic C57Bl/6 mice ST:STUDY_TYPE Insulin depravation ST:STUDY_SUMMARY Experiments were conducted using 13-wk-old male C57BL/6J mice (Jackson ST:STUDY_SUMMARY Bar Harbor, ME). Mice were housed individually with free access to water and ST:STUDY_SUMMARY (TD.10112; Harlan Laboratories, Indianapolis, IN), with a 12:12-h light-dark ST:STUDY_SUMMARY and temperature and humidity control. Mice were acclimated for 1 wk prior to ST:STUDY_SUMMARY beginning of the experiment. The protocol was approved by the Mayo Clinic ST:STUDY_SUMMARY Animal Care and Use Committee. Following a 6-h fast, mice were given ST:STUDY_SUMMARY injections of STZ (125 mg/kg; in sodium acetate buffer, pH = 4.5) (67). ST:STUDY_SUMMARY were repeated on the following day. Control animals received intraperitoneal ST:STUDY_SUMMARY of vehicle. Only mice that displayed blood glucose ?300 mg/dl and an increase ST:STUDY_SUMMARY blood ketones (both values by Precision Xtra glucometer; Abbott Laboratories, ST:STUDY_SUMMARY Park, IL), hyperphagia, and polyuria and were positive for urine glucose ST:STUDY_SUMMARY via dipstick (Uristix, Bayer, Pittsburgh, PA) on day 7 after the first STZ dose ST:STUDY_SUMMARY included in the experiment. Animals that were positive for STZ diabetes ST:STUDY_SUMMARY LinBit subcutaneous insulin implant (LinShin Canada, Toronto, ON, Canada) (79) ST:STUDY_SUMMARY pentobarbital sodium anesthesia (Nebutal, 40 mg/kg of body wt) according to the ST:STUDY_SUMMARY protocol. Each animal received two subcutaneous implants (total dose: 0.2 U/24 ST:STUDY_SUMMARY for >30 days, 10 U/kg for 20-g mice). Insulin treatment was continued for 3 wk. ST:STUDY_SUMMARY animals (C; n = 13) received blank implants. Diabetic control was confirmed by ST:STUDY_SUMMARY measurements of blood and urinary glucose. In some cases, when urine glucose ST:STUDY_SUMMARY present and blood glucose was >288 mg/dl, the animal received a third implant. ST:STUDY_SUMMARY insulin treatment was continued until initially lower plasma glucose content in ST:STUDY_SUMMARY animals reached control values. Three weeks following implantation, diabetic ST:STUDY_SUMMARY were divided randomly into diabetic-treated (D + I; n = 13) and ST:STUDY_SUMMARY (D ? I; n = 13) groups. Insulin implants were removed from the D ? I group ST:STUDY_SUMMARY pentobarbital anesthesia, which led to the return of the diabetic phenotype ST:STUDY_SUMMARY 24 h. Animals from the D + I group continued on insulin treatment (Fig. 1). At ST:STUDY_SUMMARY age of 18 wk, animals from all groups were analyzed for body composition by an ST:STUDY_SUMMARY Body Composition Analyzer (EchoMRI, Houston, TX) and euthanized by decapitation ST:STUDY_SUMMARY wk after the initial STZ or vehicle dose. Figure 1 depicts the timeline of the ST:STUDY_SUMMARY and blood glucose profiles for each experimental group. Additional animals were ST:STUDY_SUMMARY for estimation of skeletal muscle insulin sensitivity by acute insulin ST:STUDY_SUMMARY The mice were divided into the C (n = 6), D ? I (n = 7), and D + I (n = 7) ST:STUDY_SUMMARY and followed appropriate experimental treatment, except for acute insulin ST:STUDY_SUMMARY 10 min prior to euthanization by pentobarbital overdose. Figure 1 of the ST:STUDY_SUMMARY PDF of the article summarizes the study design ST:INSTITUTE Mayo Clinic ST:DEPARTMENT Endocrinology ST:LAST_NAME Nair ST:FIRST_NAME Sreekumaran ST:ADDRESS - ST:EMAIL Dasari.Surendra@mayo.edu ST:PHONE - ST:NUM_GROUPS 3 ST:TOTAL_SUBJECTS 39 #SUBJECT SU:SUBJECT_TYPE Animal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENOTYPE_STRAIN C57BL/6J SU:AGE_OR_AGE_RANGE 13-wk-old SU:GENDER Male SU:SPECIES_GROUP Mammal #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - C2 Treatment:control SUBJECT_SAMPLE_FACTORS - C3 Treatment:control SUBJECT_SAMPLE_FACTORS - C35 Treatment:control SUBJECT_SAMPLE_FACTORS - C38 Treatment:control SUBJECT_SAMPLE_FACTORS - C4 Treatment:control SUBJECT_SAMPLE_FACTORS - C44 Treatment:control SUBJECT_SAMPLE_FACTORS - C45 Treatment:control SUBJECT_SAMPLE_FACTORS - C47 Treatment:control SUBJECT_SAMPLE_FACTORS - C48 Treatment:control SUBJECT_SAMPLE_FACTORS - C49 Treatment:control SUBJECT_SAMPLE_FACTORS - C5 Treatment:control SUBJECT_SAMPLE_FACTORS - C50 Treatment:control SUBJECT_SAMPLE_FACTORS - C51 Treatment:control SUBJECT_SAMPLE_FACTORS - C6 Treatment:control SUBJECT_SAMPLE_FACTORS - D+14 Treatment:diabetic treated SUBJECT_SAMPLE_FACTORS - D+21 Treatment:diabetic treated SUBJECT_SAMPLE_FACTORS - D+23 Treatment:diabetic treated SUBJECT_SAMPLE_FACTORS - D+24 Treatment:diabetic treated SUBJECT_SAMPLE_FACTORS - D+32 Treatment:diabetic treated SUBJECT_SAMPLE_FACTORS - D+52 Treatment:diabetic treated SUBJECT_SAMPLE_FACTORS - D+53 Treatment:diabetic treated SUBJECT_SAMPLE_FACTORS - D+54 Treatment:diabetic treated SUBJECT_SAMPLE_FACTORS - D+55 Treatment:diabetic treated SUBJECT_SAMPLE_FACTORS - D+56 Treatment:diabetic treated SUBJECT_SAMPLE_FACTORS - D+57 Treatment:diabetic treated SUBJECT_SAMPLE_FACTORS - D+58 Treatment:diabetic treated SUBJECT_SAMPLE_FACTORS - D+70 Treatment:diabetic treated SUBJECT_SAMPLE_FACTORS - D-25 Treatment:diabetic untreated SUBJECT_SAMPLE_FACTORS - D-28 Treatment:diabetic untreated SUBJECT_SAMPLE_FACTORS - D-31 Treatment:diabetic untreated SUBJECT_SAMPLE_FACTORS - D-33 Treatment:diabetic untreated SUBJECT_SAMPLE_FACTORS - D-36 Treatment:diabetic untreated SUBJECT_SAMPLE_FACTORS - D-39 Treatment:diabetic untreated SUBJECT_SAMPLE_FACTORS - D-40 Treatment:diabetic untreated SUBJECT_SAMPLE_FACTORS - D-41 Treatment:diabetic untreated SUBJECT_SAMPLE_FACTORS - D-42 Treatment:diabetic untreated SUBJECT_SAMPLE_FACTORS - D-43 Treatment:diabetic untreated SUBJECT_SAMPLE_FACTORS - D-60 Treatment:diabetic untreated SUBJECT_SAMPLE_FACTORS - D-61 Treatment:diabetic untreated SUBJECT_SAMPLE_FACTORS - D-63 Treatment:diabetic untreated SUBJECT_SAMPLE_FACTORS - D-64 Treatment:diabetic untreated SUBJECT_SAMPLE_FACTORS - D-67 Treatment:diabetic untreated SUBJECT_SAMPLE_FACTORS - D-68 Treatment:diabetic untreated SUBJECT_SAMPLE_FACTORS - D-69 Treatment:diabetic untreated #COLLECTION CO:COLLECTION_SUMMARY Mitochondria were isolated from quadriceps muscle by differential CO:COLLECTION_SUMMARY as described previously (38). Briefly, quadriceps muscle samples were CO:COLLECTION_SUMMARY on ice using a motor-driven Potter-Elvehjem tissue grinder. After initial CO:COLLECTION_SUMMARY the supernatant containing the mitochondrial and sarcoplasmic fraction was CO:COLLECTION_SUMMARY to a chilled microcentrifuge tube and centrifuged at 10,000 g for 2 min to CO:COLLECTION_SUMMARY mitochondria. The supernatant containing sarcoplasmic fraction was frozen for CO:COLLECTION_SUMMARY analysis. Mitochondrial pellet was washed twice by resuspending/centrifugation CO:COLLECTION_SUMMARY finally suspended in a mitochondrial storage buffer. The levels of both the CO:COLLECTION_SUMMARY and sphingolipids in homogenates and various muscle fractions were normalized CO:COLLECTION_SUMMARY total protein content, as measured by 660 nm Protein Assay (Thermo Scientific; CO:COLLECTION_SUMMARY Protein Biology Products, Rockford, IL). #TREATMENT TR:TREATMENT_SUMMARY Control/Diabetic; insulin treated/Diabetic; insulin deprived TR:TREATMENT_COMPOUND blank/Insulin/Insulin TR:TREATMENT_ROUTE Skin implants TR:ANIMAL_ANESTHESIA phenobarbital TR:ANIMAL_ENDP_EUTHANASIA 5 weeks after treatment TR:ANIMAL_ENDP_TISSUE_COLL_LIST plasma, muscle, liver and skin #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Mitochondria were isolated from quadriceps muscle by differential SP:SAMPLEPREP_SUMMARY as described previously (38). Briefly, quadriceps muscle samples were SP:SAMPLEPREP_SUMMARY on ice using a motor-driven Potter-Elvehjem tissue grinder. After initial SP:SAMPLEPREP_SUMMARY the supernatant containing the mitochondrial and sarcoplasmic fraction was SP:SAMPLEPREP_SUMMARY to a chilled microcentrifuge tube and centrifuged at 10,000 g for 2 min to SP:SAMPLEPREP_SUMMARY mitochondria. The supernatant containing sarcoplasmic fraction was frozen for SP:SAMPLEPREP_SUMMARY analysis. Mitochondrial pellet was washed twice by resuspending/centrifugation SP:SAMPLEPREP_SUMMARY finally suspended in a mitochondrial storage buffer. The levels of both the SP:SAMPLEPREP_SUMMARY and sphingolipids in homogenates and various muscle fractions were normalized SP:SAMPLEPREP_SUMMARY total protein content, as measured by 660 nm Protein Assay (Thermo Scientific; SP:SAMPLEPREP_SUMMARY Protein Biology Products, Rockford, IL). / Plasma free fatty acid SP:SAMPLEPREP_SUMMARY were measured by liquid chromatography/mass spectrometry (LC/MS), as described SP:SAMPLEPREP_SUMMARY (51). Briefly, 50 ?l of plasma was spiked with heptadecanoate internal standard SP:SAMPLEPREP_SUMMARY and analyzed with Applied Biosystems (Foster City, CA) API5000 mass SP:SAMPLEPREP_SUMMARY coupled with a Cohesive (Franklin, MA) TX2 liquid chromatography system. SP:SAMPLEPREP_SUMMARY of individual FFA was measured against a six-point standard curve prepared for SP:SAMPLEPREP_SUMMARY analyte. Both the ISTD and individual FFA standard curves were prepared in 2% SP:SAMPLEPREP_SUMMARY acid-free human albumin solution. All analytes were monitored as their [M ? H]? SP:SAMPLEPREP_SUMMARY plasma LCFa-CoA esters were estimated using the LC-MS/MS method (9). After SP:SAMPLEPREP_SUMMARY in the presence of internal standard (20 ng of heptadecanoyl-CoA), samples were SP:SAMPLEPREP_SUMMARY by UHPLC-ESI-MS/MS operating in multiple reaction monitoring mode [Waters SP:SAMPLEPREP_SUMMARY UHPLC, C8 UPLC BEH column 2.1 × 150 mm, 1.7 ?m (Waters, Milford, MA) and TSQ SP:SAMPLEPREP_SUMMARY Ultra triple-quad mass spectrometer (Thermo Fisher Scientific, Waltham, MA)]. SP:SAMPLEPREP_SUMMARY standard curves were prepared using chemicals from Avanti Polar Lipids. SP:SAMPLEPREP_PROTOCOL_FILENAME PMID-24368672-Zabielski-Nair-AJPEM-2014.pdf SP:SAMPLEPREP_PROTOCOL_COMMENTS Pubmed ID: 24368672 #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Plasma free fatty acid concentrations were measured by liquid CH:CHROMATOGRAPHY_SUMMARY spectrometry (LC/MS), as described previously (51). Briefly, 50 ?l of plasma CH:CHROMATOGRAPHY_SUMMARY spiked with heptadecanoate internal standard (ISTD) and analyzed with Applied CH:CHROMATOGRAPHY_SUMMARY (Foster City, CA) API5000 mass spectrometer coupled with a Cohesive (Franklin, CH:CHROMATOGRAPHY_SUMMARY TX2 liquid chromatography system. Concentration of individual FFA was measured CH:CHROMATOGRAPHY_SUMMARY a six-point standard curve prepared for each analyte. Both the ISTD and CH:CHROMATOGRAPHY_SUMMARY FFA standard curves were prepared in 2% fatty acid-free human albumin solution. CH:CHROMATOGRAPHY_SUMMARY analytes were monitored as their [M ? H]? ions. LCFa-CoA esters were estimated CH:CHROMATOGRAPHY_SUMMARY the LC-MS/MS method (9). After extraction in the presence of internal standard CH:CHROMATOGRAPHY_SUMMARY ng of heptadecanoyl-CoA), samples were analyzed by UHPLC-ESI-MS/MS operating in CH:CHROMATOGRAPHY_SUMMARY reaction monitoring mode [Waters Acquity UHPLC, C8 UPLC BEH column 2.1 150 mm, CH:CHROMATOGRAPHY_SUMMARY ?m (Waters, Milford, MA) and TSQ Quantum Ultra triple-quad mass spectrometer CH:CHROMATOGRAPHY_SUMMARY Fisher Scientific, Waltham, MA)]. All standard curves were prepared using CH:CHROMATOGRAPHY_SUMMARY from Avanti Polar Lipids. CH:CHROMATOGRAPHY_TYPE - CH:INSTRUMENT_NAME - CH:COLUMN_NAME - CH:METHODS_FILENAME PMID-24368672-Zabielski-Nair-AJPEM-2014.pdf CH:CHROMATOGRAPHY_COMMENTS Pubmed ID: 24368672 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME ABI Sciex API 5000 QQQ MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS Briefly, 50 ul of plasma was spiked with heptadecanoate internal standard MS:MS_COMMENTS and analyzed with Applied Biosystems (Foster City, CA) API5000 mass MS:MS_COMMENTS coupled with a Cohesive (Franklin, MA) TX2 liquid chromatography system. MS:MS_COMMENTS of individual FFA was measured against a six-point standard curve prepared for MS:MS_COMMENTS analyte. Both the ISTD and individual FFA standard curves were prepared in 2% MS:MS_COMMENTS acid-free human albumin solution. All analytes were monitored as their [M - H]- MS:MS_COMMENTS API5000 mass spectrometer coupled with a Cohesive (Franklin, MA) TX2 liquid MS:MS_COMMENTS system #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS uM MS_METABOLITE_DATA_START Samples C3 C44 C45 C47 C48 C49 C5 C50 C51 D+14 D+24 D+52 D+53 D+54 D+55 D+56 D+57 D+58 D+70 D-25 D-28 D-31 D-36 D-41 D-42 D-43 D-60 D-61 D-63 D-67 D-69 Factors Treatment:control Treatment:control Treatment:control Treatment:control Treatment:control Treatment:control Treatment:control Treatment:control Treatment:control Treatment:diabetic treated Treatment:diabetic treated Treatment:diabetic treated Treatment:diabetic treated Treatment:diabetic treated Treatment:diabetic treated Treatment:diabetic treated Treatment:diabetic treated Treatment:diabetic treated Treatment:diabetic treated Treatment:diabetic untreated Treatment:diabetic untreated Treatment:diabetic untreated Treatment:diabetic untreated Treatment:diabetic untreated Treatment:diabetic untreated Treatment:diabetic untreated Treatment:diabetic untreated Treatment:diabetic untreated Treatment:diabetic untreated Treatment:diabetic untreated Treatment:diabetic untreated arachidonic acid 13.8504 13.3339 12.2160 10.8482 13.5054 10.5501 13.6533 12.4493 16.2113 16.9930 6.5686 16.2419 15.1135 1.9693 15.2873 13.3541 12.1273 12.4994 11.5342 4.8251 5.0795 2.9801 12.2394 12.5714 4.2261 18.6938 13.0847 18.0556 14.9386 16.0896 19.1486 elaidic acid 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 linoleic acid 244.2618 327.4403 258.2500 223.4178 182.9635 219.3733 205.8335 167.4312 304.2726 320.8385 59.9321 240.1323 287.1508 71.1733 269.3981 290.5246 204.0629 157.3808 215.2354 131.2519 82.2469 47.0276 303.1033 335.5392 40.2195 487.7377 337.3508 624.8732 287.2641 889.4591 666.0784 linolenic acid 42.8341 47.4301 35.7795 30.7379 25.2128 33.4660 30.5086 23.3406 52.6265 48.5876 6.9452 28.8091 34.2258 9.9814 33.4688 38.1802 27.2320 20.7955 23.3257 12.8187 12.3449 5.7998 44.0953 49.4155 4.5180 79.3517 46.6674 91.7382 37.2213 186.7068 110.7009 myristic acid 9.6376 26.6749 24.2911 7.6783 7.2832 9.7653 5.5657 3.0074 11.4413 5.8021 1.0127 6.8653 5.1248 1.4879 5.3674 4.9654 1.1711 2.0724 3.5402 4.7377 3.3782 0.0000 2.0652 2.2932 1.0198 3.9620 1.2086 9.3920 2.2836 7.9058 9.5126 oleic acid 119.4748 158.7289 131.8964 86.2526 76.7325 84.2217 93.8793 61.3628 139.5115 122.3238 33.0740 104.8827 116.5999 30.9407 119.4648 133.6590 78.0439 66.4842 78.0066 57.8008 38.3290 13.4841 99.5273 114.8568 11.2064 161.4988 116.7647 246.1194 100.9013 402.8514 295.9707 palmitic acid 162.0090 275.5816 240.5376 180.6636 176.5379 180.0336 149.5223 142.6934 258.6914 194.4096 47.5372 168.0452 205.7764 65.4579 192.6724 168.6953 109.3017 114.9930 130.9655 74.5386 53.4533 22.6754 148.6053 172.5051 22.8115 242.7654 167.0698 267.7250 165.0202 358.1991 307.5975 palmitoleic acid 19.4875 44.7234 49.2704 19.5544 18.8268 19.4963 17.7176 10.5233 37.1961 12.9088 4.1597 17.5860 15.0425 6.7232 16.6555 14.8441 9.6335 7.8033 11.0202 9.9724 5.9078 1.3072 6.0997 7.7542 1.4335 9.4432 6.8006 20.3802 8.9928 13.5492 22.7240 stearic acid 56.5978 76.7075 59.7160 54.3215 53.1829 48.6920 41.3317 47.7264 65.3180 91.9387 17.9384 52.8159 82.2845 25.4363 63.0850 52.0892 42.3182 43.7023 51.9152 20.9702 18.3048 6.2993 73.0351 75.2052 6.6917 112.6918 81.1077 115.1512 79.3176 133.6531 113.3471 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name moverz_quant ri ri_type pubchem_id inchi_key kegg_id other_id other_id_type arachidonic acid MAYO_ID elaidic acid MAYO_ID linoleic acid MAYO_ID linolenic acid MAYO_ID myristic acid MAYO_ID oleic acid MAYO_ID palmitic acid MAYO_ID palmitoleic acid MAYO_ID stearic acid MAYO_ID METABOLITES_END #END