#METABOLOMICS WORKBENCH Tomoya_Tsukimi_20170309_001205 DATATRACK_ID:861 STUDY_ID:ST000570 ANALYSIS_ID:AN000877 PROJECT_ID:PR000418 VERSION 1 CREATED_ON March 13, 2017, 11:57 am #PROJECT PR:PROJECT_TITLE Metabolome analysis of the cecal contents of GF mice and GF mice colonized with PR:PROJECT_TITLE dominant gut microbes present in the ceca of neonatal and adult mice PR:PROJECT_SUMMARY The high susceptibility of neonates to infections has been assumed to be due to PR:PROJECT_SUMMARY immaturity of the immune system, but the mechanism remains unclear. By PR:PROJECT_SUMMARY colonizing adult germ-free mice with the cecal contents of neonatal and adult PR:PROJECT_SUMMARY mice, we show that the neonatal microbiota is impaired in mediating colonization PR:PROJECT_SUMMARY resistance against two major pathogens causing mortality in neonates. The lack PR:PROJECT_SUMMARY of colonization resistance was caused by the absence of Clostridiales in the PR:PROJECT_SUMMARY neonatal microbiota. Administration of Clostridiales, but not Bacteroidales, PR:PROJECT_SUMMARY restored colonization resistance and abrogated intestinal pathology upon PR:PROJECT_SUMMARY pathogen challenge. Conversely, depletion of Clostridiales abolished PR:PROJECT_SUMMARY colonization resistance in adult mice. Furthermore, intragastric administration PR:PROJECT_SUMMARY of Clostridiales protected neonatal mice from pathogen infection. The neonatal PR:PROJECT_SUMMARY bacteria enhanced the ability of these protective Clostridiales to colonize the PR:PROJECT_SUMMARY gut. These results identify the gut microbiota as a critical determinant of PR:PROJECT_SUMMARY increased susceptibility to enteric infection during the neonatal period. PR:INSTITUTE Keio University PR:DEPARTMENT Institute for Advanced Biosciences PR:LAST_NAME Fukuda PR:FIRST_NAME Shinji PR:ADDRESS Tsuruoka, Yamagata 997-0052, Japan PR:EMAIL sfukuda@sfc.keio.ac.jp PR:PHONE +81-235-29-0528 #STUDY ST:STUDY_TITLE Metabolome analysis of the cecal contents of GF mice and GF mice colonized with ST:STUDY_TITLE dominant gut microbes present in the ceca of neonatal and adult mice ST:STUDY_SUMMARY Metabolome profiles of GF or GF mice reconstituted with Esherichia coli (EC), ST:STUDY_SUMMARY Bacteroides acidifaciens (Bac), or Clostridia consortium (CL) were compared. ST:INSTITUTE Keio University ST:DEPARTMENT Institute for Advanced Biosciences ST:LAST_NAME Fukuda ST:FIRST_NAME Shinji ST:ADDRESS Tsuruoka, Yamagata 997-0052, Japan ST:EMAIL sfukuda@sfc.keio.ac.jp ST:PHONE +81-235-29-0528 ST:NUM_GROUPS 4 ST:TOTAL_SUBJECTS 17 #SUBJECT SU:SUBJECT_TYPE Animal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENOTYPE_STRAIN C57BL/6 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - 1_B1 colonized bacteria:Bacteroides acidifaciens SUBJECT_SAMPLE_FACTORS - 1_B2 colonized bacteria:Bacteroides acidifaciens SUBJECT_SAMPLE_FACTORS - 1_B3 colonized bacteria:Bacteroides acidifaciens SUBJECT_SAMPLE_FACTORS - 1_B4 colonized bacteria:Bacteroides acidifaciens SUBJECT_SAMPLE_FACTORS - 1_B5 colonized bacteria:Bacteroides acidifaciens SUBJECT_SAMPLE_FACTORS - 1_CL1 colonized bacteria:Clostridia SUBJECT_SAMPLE_FACTORS - 1_CL2 colonized bacteria:Clostridia SUBJECT_SAMPLE_FACTORS - 1_CL3 colonized bacteria:Clostridia SUBJECT_SAMPLE_FACTORS - 1_CL4 colonized bacteria:Clostridia SUBJECT_SAMPLE_FACTORS - 1_CL5 colonized bacteria:Clostridia SUBJECT_SAMPLE_FACTORS - 2_EB1 colonized bacteria:Esherichia coli SUBJECT_SAMPLE_FACTORS - 2_EB2 colonized bacteria:Esherichia coli SUBJECT_SAMPLE_FACTORS - 2_EB3 colonized bacteria:Esherichia coli SUBJECT_SAMPLE_FACTORS - 5_C1 colonized bacteria:none (GF) SUBJECT_SAMPLE_FACTORS - 5_C2 colonized bacteria:none (GF) SUBJECT_SAMPLE_FACTORS - 5_C3 colonized bacteria:none (GF) SUBJECT_SAMPLE_FACTORS - 5_C4 colonized bacteria:none (GF) #COLLECTION CO:COLLECTION_SUMMARY Feces of GF or GF mice reconstituted with Esherichia coli (EC), Bacteroides CO:COLLECTION_SUMMARY acidifaciens (Bac), or Clostridia consortium (CL) CO:SAMPLE_TYPE Feces CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY Animals Specific-pathogen-free (SPF) C57BL/6 mice were originally purchased from TR:TREATMENT_SUMMARY Jackson Laboratories. Wild-type mice on GF background were bred and maintained TR:TREATMENT_SUMMARY at the Germ-free Animal Core Facility of the University of Michigan. GF mice TR:TREATMENT_SUMMARY were maintained in flexible film isolators and were checked weekly for germ-free TR:TREATMENT_SUMMARY status by aerobic and anaerobic culture. The absence of microbiota was verified TR:TREATMENT_SUMMARY by microscopic analysis of stained cecal contents to detect unculturable TR:TREATMENT_SUMMARY contamination. All animal studies were performed according to approved protocols TR:TREATMENT_SUMMARY by the University of Michigan Committee on the Use and Care of Animals. TR:TREATMENT_SUMMARY Reconstitution of GF mice with cecal contents and defined bacteria Cecal TR:TREATMENT_SUMMARY contents from single neonatal and 7-week old SPF mice were diluted into 50 ml TR:TREATMENT_SUMMARY (per one adult mouse) or 5 ml (per one 4, 12, 16-day old mouse) of PBS, and TR:TREATMENT_SUMMARY passed through a 70 μm cell strainer to eliminate clumps and debris under TR:TREATMENT_SUMMARY sterile conditions) under an atmosphere of 5% H2, 5% CO2 and 90% N2 in an TR:TREATMENT_SUMMARY anaerobic growth chamber (Coy Manufacturing, Grass Lake, MI). Then 200 μl of TR:TREATMENT_SUMMARY the suspension was processed by intragastric gavage into each of the recipient TR:TREATMENT_SUMMARY GF mice. Bacteroides species were anaerobically grown at 37°C overnight in TR:TREATMENT_SUMMARY pre-reduced chopped-meat broth (Anaerobe systems, Morgan Hill, CA). E. coli were TR:TREATMENT_SUMMARY grown aerobically overnight in LB broth with shaking. TR:TREATMENT_PROTOCOL_FILENAME Treatment_Protocol.pdf TR:TREATMENT Biotic TR:TREATMENT_COMPOUND Suspension of diluted cecal contents TR:TREATMENT_ROUTE Intragastric gavage TR:TREATMENT_DOSE 200 μl #SAMPLEPREP SP:SAMPLEPREP_SUMMARY CE-TOFMS-based metabolome analysis(sample preparation) Fecal samples were freeze SP:SAMPLEPREP_SUMMARY dried, and disrupted by vigorous shaking at 1,500 rpm for 10 min with four 3 mm SP:SAMPLEPREP_SUMMARY zirconia beads by Shake Master NEO (Bio Medical Science Inc.). 10 mg (±0.5 mg) SP:SAMPLEPREP_SUMMARY fecal samples were homogenized with 500 μl MeOH containing internal standards SP:SAMPLEPREP_SUMMARY (20 μM each of methionine sulfone, and D-camphor-10-sulfonic acid (CSA)) and SP:SAMPLEPREP_SUMMARY 100 mg of 0.1 mm and four of 3 mm zirconia/silica beads (BioSpec Products). SP:SAMPLEPREP_SUMMARY After vigorous shaking (1,500 rpm for 5 min) by Shake Master NEO (Bio Medical SP:SAMPLEPREP_SUMMARY Science Inc.), 200 μl of Milli-Q water and 500 μl of chloroform was added and SP:SAMPLEPREP_SUMMARY then shaking in a same manner as before. After centrifugation at 4,600 × g for SP:SAMPLEPREP_SUMMARY 15 min at 4°C, the supernatant was transferred to a 5kDa cutoff centrifugal SP:SAMPLEPREP_SUMMARY filter tube. The filtrate was centrifugally concentrated at 40°C and SP:SAMPLEPREP_SUMMARY reconstituted with 40 μl of Milli-Q water. SP:SAMPLEPREP_PROTOCOL_FILENAME Sample_Prep_Protocol.pdf SP:PROCESSING_METHOD Homogenized with 500 μl MeOH containing internal standards and 100 mg of 0.1 mm SP:PROCESSING_METHOD and four of 3 mm zirconia/silica beads SP:PROCESSING_STORAGE_CONDITIONS On ice SP:EXTRACTION_METHOD Chloroform SP:EXTRACT_CLEANUP 5kDa cutoff centrifugal filter tube SP:SAMPLE_RESUSPENSION 40 μl of Milli-Q water #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE CE CH:INSTRUMENT_NAME Agilent CE CH:COLUMN_NAME COSMO(+), i.d.50 µm(anion), Fused-silica, i.d. 50 µm(cation) CH:INTERNAL_STANDARD 20 μM each of methionine sulfone, and D-camphor-10-sulfonic acid (CSA) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME Agilent CE-TOFMS MS:INSTRUMENT_TYPE TOF MS:MS_TYPE ESI MS:ION_MODE POSITIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS nmol/g MS_METABOLITE_DATA_START Samples 1_B1 1_B2 1_B3 1_B4 1_B5 1_CL1 1_CL2 1_CL3 1_CL4 1_CL5 2_EB1 2_EB2 2_EB3 5_C1 5_C2 5_C3 5_C4 Factors colonized bacteria:Bacteroides acidifaciens colonized bacteria:Bacteroides acidifaciens colonized bacteria:Bacteroides acidifaciens colonized bacteria:Bacteroides acidifaciens colonized bacteria:Bacteroides acidifaciens colonized bacteria:Clostridia colonized bacteria:Clostridia colonized bacteria:Clostridia colonized bacteria:Clostridia colonized bacteria:Clostridia colonized bacteria:Esherichia coli colonized bacteria:Esherichia coli colonized bacteria:Esherichia coli colonized bacteria:none (GF) colonized bacteria:none (GF) colonized bacteria:none (GF) colonized bacteria:none (GF) Acetyl CoA 4 6 7 7 7 151 213 22 83 45 Adipate 24 95 12 32 31 18 21 41 49 19 14 10 29 122 103 49 71 ADP 6 4 11 5 7 52 88 14 21 13 4 5 AMP 185 151 88 106 Azelate 16 17 13 13 11 191 155 191 313 118 12 15 19 10 10 22 21 Butanoate 4915 2072 1255 2717 2132 Carbamoylaspartate 8 11 6 6 6 26 12 5 11 13 3 Cholate 94 62 93 206 94 257 107 92 135 cis-Aconitate 108 537 12 425 310 31 62 186 201 9 89 17 194 879 1191 426 350 Citraconate 5 20 3 12 7 7 5 14 21 5 7 5 14 21 24 16 20 Citramalate 36 35 12 35 31 27 31 129 45 56 11 9 13 41 31 23 25 Citrate 1486 4053 385 2615 3586 980 403 1176 1219 262 805 434 1544 2393 16312 2578 1673 CMP 4 4 7 4 3 34 43 29 32 12 3 2 4 Cysteate 19 21 28 27 21 205 313 53 200 135 7 11 11 13 16 14 22 Cysteine S-sulfate 32 31 17 22 18 3 21 7 10 211 189 291 209 285 237 712 Dihydroorotate 6 Dodecanedioate 6 12 25 10 dTMP 24 25 15 18 12 F6P 5 9 12 9 222 82 21 148 56 Fumarate 299 107 159 165 140 18 10 192 9 150 101 147 13 26 27 16 G1P 169 47 16 48 22 G6P 15 34 51 53 27 902 350 90 713 238 4 5 4 Gluconate 1703 1814 1953 2223 1813 133 75 1296 2897 1211 1777 Glucosamine 6-phosphate 15 Glucuronate 92 153 167 886 1216 911 830 Glutarate 74 484 18 100 109 91 160 189 193 130 79 41 192 331 337 188 468 Glycerate 228 155 200 270 263 336 389 626 271 314 142 139 109 167 200 129 230 Glycerophosphate 55 61 26 36 29 386 310 145 305 109 17 31 115 91 38 18 12 Glycolate 763 896 3256 3211 1267 17952 1224 662 1355 852 707 Hippurate 72 32 10 31 38 61 118 56 35 Homovanillate 3699 2546 1168 2922 2861 IMP 25 Isethionate 75 259 21 224 140 94 99 233 181 110 44 45 83 903 774 352 259 Isobutyryl CoA 96 42 5 74 37 Isocitrate 64 98 13 141 105 28 56 167 40 9 37 26 96 389 628 184 127 Itaconate 7 8 6 9 6 13 9 5 9 7 3 5 4 3 Lactate 1916 1822 2286 1600 1961 23912 9245 12217 75807 14113 1154 1763 1770 7435 13270 17598 13299 Malate 1563 863 993 1121 877 499 81 110 302 68 312 311 322 106 695 175 130 Malonate 141 116 70 106 90 376 401 548 320 322 101 96 183 130 160 94 157 Methanesulfonate 41 59 128 55 Methyl sulfate 25 29 18 21 19 16 14 13 23 15 14 15 17 26 33 15 20 Mucate 45 60 62 72 51 11 4 19 6 11 14 19 87 130 70 85 N-Acetylaspartate 31 55 15 38 34 1215 874 85 721 521 12 6 15 30 102 30 24 N-Acetyl-beta-alanine 142 134 138 175 134 32 28 49 35 31 248 296 309 153 237 136 240 N-Acetylglucosamine 1-phosphate 3 4 5 6 178 170 112 190 119 N-Acetylglucosamine 6-phosphate 8 12 15 14 10 79 67 70 66 5 N-Acetylglutamate 19 59 18 45 34 420 340 125 331 198 31 30 42 31 111 36 25 NAD+ 947 963 450 NADP+ 32 39 N-Carbamylglutamate 13 40 N-Formylaspartate 79 Nicotinate 403 436 347 383 238 39 26 44 o-Hydroxyhippurate 16 O-Phosphoserine 10 13 7 43 56 11 31 Orotate 56 53 40 58 52 44 98 44 36 24 39 29 49 69 78 38 39 Pentanoate 6 17 8 49 52 25 9 6 9 7 10 13 10 Phenylpyruvate 49 207 314 47 91 Phthalate 6 4 4 5 5 13 11 12 18 9 4 5 5 4 5 5 7 p-Hydroxybenzoate 41 p-Hydroxyphenylacetate 72 147 108 141 136 Pimelate 7 35 5 21 16 17 21 29 37 12 3 2 7 26 55 24 23 Propionate 313 232 237 141 94 212 Pyruvate 58 106 147 265 293 235 229 979 283 198 Quinate 596 602 821 812 736 1049 1119 653 889 725 617 1117 695 858 Ru5P 84 76 7 114 75 S7P 8 12 17 16 11 158 103 25 149 54 Saccharate 84 89 103 118 75 189 319 169 180 Sebacate 1 1 2 1 1 20 23 20 17 12 1 1 2 1 1 1 2 Shikimate 443 500 605 672 546 194 462 765 480 570 Succinate 9876 6803 7728 7497 6008 1333 1186 1060 2015 745 1461 1220 2257 349 1706 484 319 Taurocholate 146 315 168 212 276 142 250 395 308 350 556 1024 2891 87 646 834 4091 Terephthalate 4 4 7 7 4 4 4 4 4 5 5 4 24 2 3 4 4 Threonate 336 688 373 580 408 209 264 387 275 194 179 230 289 626 684 358 392 trans-4-Hydroxy-3-methoxycinnamate 78 58 68 59 46 18 22 97 75 89 82 30 54 31 46 trans-Aconitate 8 96 53 13 19 11 19 76 20 5 8 149 163 92 50 UDP 3 4 6 6 4 29 38 6 13 5 UDP-glucuronate 12 UDP-N-acetylglucosamine 38 52 UMP 6 7 12 12 8 109 105 42 60 15 Urate 137 200 138 197 156 65 100 162 243 22 16 19 39 504 520 212 185 XMP 18 15 4 12 2,3-Pyridinedicarboxylate 11 17 9 12 11 6 11 12 6 10 9 10 9 16 25 11 16 2,5-Dihydroxybenzoate 17 32 11 24 19 377 268 2-Deoxyglucose 6-phosphate 24 20 5 31 16 2-Hydroxybutyrate 5 5 6 34 16 22 64 20 9 12 14 9 42 14 12 2-Hydroxyglutarate 161 189 112 132 207 442 564 455 476 264 17 22 39 164 54 58 2-Hydroxyisobutyrate 3 9 3 4 6 7 13 17 8 7 2-Hydroxypentanoate 37 58 31 56 45 47 45 69 30 48 30 25 33 26 68 33 44 2-Hydroxyphenylacetate 10 19 13 8 13 2-Isopropylmalate 4 5 2 4 3 2 2 4 5 2 7 5 6 4 2-Oxoadipate 14 59 35 27 21 199 1369 68 178 2-Oxoglutarate 412 1726 927 1553 717 242 1007 56 188 145 221 733 7967 444 500 2-Oxoisopentanoate 6 25 8 19 14 21 35 19 7 5 27 102 249 45 60 3-(2-Hydroxyphenyl)propionate 387 510 554 561 501 29 72 48 42 80 405 490 415 149 285 159 222 3-(4-Hydroxyphenyl)propionate 226 141 225 269 173 3-Hydroxy-3-methylglutarate 125 199 50 99 122 9 5 22 36 4 12 5 25 165 124 38 71 3-Hydroxybutyrate 27 45 25 39 39 40 46 38 30 29 17 18 27 43 93 32 48 3-Hydroxypropionate 125 3-Indoxyl sulfate 58 40 11 146 71 59 44 268 3-Methylbutanoate 8 17 22 31 25 36 20 120 28 32 8 7 16 19 31 15 35 3PG 8 4 9 11 6 133 223 40 92 26 3-Phenyllactate 325 484 592 573 526 45 142 82 81 88 232 409 433 228 351 238 277 3-Phenylpropionate 1613 1780 1926 2016 1538 143 87 163 1639 2330 1791 2074 3920 1965 2831 3-Ureidopropionate 38 306 154 93 64 127 331 245 76 34 108 566 822 331 233 4-Acetylbutyrate 11 7 9 11 9 13 12 23 9 11 9 11 12 8 14 13 11 4-Hydroxymandelate 50 65 46 4-Methyl-2-oxopentanoate 102 101 74 171 191 99 58 38 138 575 1337 319 351 4-Methylthio-2-oxobutyrate 110 4-Oxopentanoate 26 20 19 22 26 42 35 74 55 40 14 18 22 16 25 29 26 5-Oxoproline 184 367 218 325 269 195 66 90 176 60 87 104 138 290 347 173 196 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name retention index quantified m/z PubChem ID KEGG ID RefMet name Acetyl CoA C00024 Acetyl-CoA Adipate C06104 Adipic acid ADP C00008 ADP AMP C00020 AMP Azelate C08261 Butanoate C00246 Butyric acid Carbamoylaspartate C00438 Cholate C00695 Cholic acid cis-Aconitate C00417 Cis-Aconitic acid Citraconate C02226 Citraconic acid Citramalate C02614 Citramalic acid Citrate C00158 Citric acid CMP C00055 CMP Cysteate C00506 Cysteic acid Cysteine S-sulfate C05824 Cysteine-S-sulfate Dihydroorotate C00337 4,5-Dihydroorotic acid Dodecanedioate C02678 Dodecanedioic acid dTMP C00364 dTMP F6P C00085 Fructose-6-phosphate Fumarate C00122 Fumaric acid G1P C00103 Glucose-1-phosphate G6P C00092 Glucose-6-phosphate Gluconate C00257 Gluconic acid Glucosamine 6-phosphate C00352 Glucosamine-6-phosphate Glucuronate C00191 Glucuronic acid Glutarate C00489 Glutaric acid Glycerate C00258 Glyceric acid Glycerophosphate C00093 Glycolate C00160 Glycolic acid Hippurate C01586 Hippuric acid Homovanillate C05582 Homovanillic acid IMP C00130 IMP Isethionate C05123 2-Hydroxyethanesulfonate Isobutyryl CoA C00630 Isobutyryl-CoA Isocitrate C00311 Isocitric acid Itaconate C00490 Itaconic acid Lactate C00186 Lactic acid Malate C00711 Malic acid Malonate C00383 Malonic acid Methanesulfonate C11145 Methyl sulfate C02704 Methyl sulfate Mucate C00879 N-Acetylaspartate C01042 N-Acetylaspartic acid N-Acetyl-beta-alanine C01073 N-acetyl-beta-alanine N-Acetylglucosamine 1-phosphate C04501 N-acetyl-D-glucosamine-1-phosphate N-Acetylglucosamine 6-phosphate C00357 N-Acetyl-Glucosamine 6-Phosphate N-Acetylglutamate C00624 N-acetylglutamic acid NAD+ C00003 NAD+ NADP+ C00006 NADP+ N-Carbamylglutamate C05829 Carglumic acid N-Formylaspartate C01044 Nicotinate C00253 Nicotinic acid o-Hydroxyhippurate C07588 O-Phosphoserine C01005 Phosphoserine Orotate C00295 Orotic acid Pentanoate C00803 Valeric acid Phenylpyruvate C00166 Phenylpyruvic acid Phthalate C01606 Phthalic acid p-Hydroxybenzoate C00156 4-Hydroxybenzoic acid p-Hydroxyphenylacetate C00642 P-Hydroxyphenylacetic acid Pimelate C02656 Pimelic acid Propionate C00163 Propionic acid Pyruvate C00022 Pyruvic acid Quinate C00296 Quinic acid Ru5P C00199 S7P C05382 Sedoheptulose 7-phosphate Saccharate C00818 Sebacate C08277 Sebacic acid Shikimate C00493 Shikimic acid Succinate C00042 Succinic acid Taurocholate C05122 Taurocholic acid Terephthalate C06337 Terephthalic acid Threonate C01620 Threonic acid trans-4-Hydroxy-3-methoxycinnamate C01494 trans-Aconitate C02341 Trans-Aconitic acid UDP C00015 UDP UDP-glucuronate C00167 UDP-glucuronic acid UDP-N-acetylglucosamine C00043 UDP-N-acetylglucosamine UMP C00105 UMP Urate C00366 Uric acid XMP C00655 XMP 2,3-Pyridinedicarboxylate C03722 2,5-Dihydroxybenzoate C00628 Gentisic acid 2-Deoxyglucose 6-phosphate C06369 2-Hydroxybutyrate C05984 2-hydroxybutyric acid 2-Hydroxyglutarate C02630 2-hydroxyglutaric acid 2-Hydroxyisobutyrate - 2-hydroxyisobutyric acid 2-Hydroxypentanoate - 2-Hydroxyphenylacetate C05852 2-hydroxyphenylacetic acid 2-Isopropylmalate C02504 2-isopropylmalic acid 2-Oxoadipate C00322 Oxoadipic acid 2-Oxoglutarate C00026 Oxoglutaric acid 2-Oxoisopentanoate C00141 3-(2-Hydroxyphenyl)propionate C01198 3-(2-Hydroxyphenyl)propanoic acid 3-(4-Hydroxyphenyl)propionate C01744 3-Hydroxy-3-methylglutarate C03761 3-Hydroxymethylglutaric acid 3-Hydroxybutyrate C01089 beta-hydroxybutyric acid 3-Hydroxypropionate C01013 3-Indoxyl sulfate - Indoxyl sulfate 3-Methylbutanoate C08262 3PG C00197 3-Phosphoglyceric acid 3-Phenyllactate C05607 3-Phenylpropionate C05629 3-Ureidopropionate C02642 Ureidopropionic acid 4-Acetylbutyrate C02129 4-Hydroxymandelate C11527 4-Methyl-2-oxopentanoate C00233 Ketoleucine 4-Methylthio-2-oxobutyrate C01180 4-Oxopentanoate - 5-Oxoproline C01879 Pyroglutamic acid METABOLITES_END #END