#METABOLOMICS WORKBENCH LCleaver1_20200723_074008 DATATRACK_ID:2102 STUDY_ID:ST001433 ANALYSIS_ID:AN002395 PROJECT_ID:PR000984 VERSION 1 CREATED_ON July 27, 2020, 11:33 am #PROJECT PR:PROJECT_TITLE Evidence for proline utilisation by oral bacterial biofilms grown in saliva PR:PROJECT_TYPE Research study PR:PROJECT_SUMMARY Within the mouth bacteria are starved of saccharides as their main nutrient PR:PROJECT_SUMMARY source between meals and it is unclear what drives their metabolism. Previously PR:PROJECT_SUMMARY oral in vitro biofilms grown in saliva have shown proteolytic degradation of PR:PROJECT_SUMMARY salivary proteins and increased extracellular proline. Although arginine and PR:PROJECT_SUMMARY glucose have been shown before to have an effect on oral biofilm growth and PR:PROJECT_SUMMARY activity, there is limited evidence for proline. Nuclear magnetic resonance PR:PROJECT_SUMMARY (NMR) spectroscopy was used to identify extracellular metabolites produced by PR:PROJECT_SUMMARY bacteria in oral biofilms grown on hydroxyapatite discs. Biofilms were PR:PROJECT_SUMMARY inoculated with whole mouth saliva and then grown for 7 days using sterilised PR:PROJECT_SUMMARY whole mouth saliva supplemented with proline, arginine and glucose as a PR:PROJECT_SUMMARY growth-medium. Overall proline had a beneficial effect on biofilm growth – PR:PROJECT_SUMMARY with significantly fewer dead bacteria present by biomass and surface area of PR:PROJECT_SUMMARY the biofilms (p <0.05). Where arginine and glucose significantly increased and PR:PROJECT_SUMMARY decreased pH, respectively, the pH of proline supplemented biofilms remained PR:PROJECT_SUMMARY neutral at pH 7.3-7.5. SDS-polyacrylamide gel electrophoresis of the spent PR:PROJECT_SUMMARY saliva from proline and arginine supplemented biofilms showed inhibition of PR:PROJECT_SUMMARY salivary protein degradation of immature biofilms. NMR analysis of the spent PR:PROJECT_SUMMARY saliva revealed that proline supplemented biofilms were metabolically similar to PR:PROJECT_SUMMARY unsupplemented biofilms, but these biofilms actively metabolised proline to PR:PROJECT_SUMMARY 5-aminopentanoate, butyrate and propionate, and actively utilised glycine. This PR:PROJECT_SUMMARY study shows that in a nutrient limited environment, proline has a beneficial PR:PROJECT_SUMMARY effect on in vitro oral biofilms grown from a saliva inoculum. PR:INSTITUTE King's College London PR:DEPARTMENT Centre for Host Microbiome Interactions PR:LAST_NAME Cleaver PR:FIRST_NAME Leanne PR:ADDRESS Floor 17, Tower Wing, Guy's Hospital, Great Maze Pond, London, London, SE1 9RT, PR:ADDRESS United Kingdom PR:EMAIL leanne.cleaver@kcl.ac.uk PR:PHONE 07464626438 PR:FUNDING_SOURCE This work was funded by a BBSRC Lido-associated PhD studentship for LC in PR:FUNDING_SOURCE association with Colgate-Palmolive, USA. #STUDY ST:STUDY_TITLE Evidence for proline utilisation by oral bacterial biofilms grown in saliva ST:STUDY_TYPE Research study ST:STUDY_SUMMARY Within the mouth bacteria are starved of saccharides as their main nutrient ST:STUDY_SUMMARY source between meals and it is unclear what drives their metabolism. Previously ST:STUDY_SUMMARY oral in vitro biofilms grown in saliva have shown proteolytic degradation of ST:STUDY_SUMMARY salivary proteins and increased extracellular proline. Although arginine and ST:STUDY_SUMMARY glucose have been shown before to have an effect on oral biofilm growth and ST:STUDY_SUMMARY activity, there is limited evidence for proline. Nuclear magnetic resonance ST:STUDY_SUMMARY (NMR) spectroscopy was used to identify extracellular metabolites produced by ST:STUDY_SUMMARY bacteria in oral biofilms grown on hydroxyapatite discs. Biofilms were ST:STUDY_SUMMARY inoculated with whole mouth saliva and then grown for 7 days using sterilised ST:STUDY_SUMMARY whole mouth saliva supplemented with proline, arginine and glucose as a ST:STUDY_SUMMARY growth-medium. Overall proline had a beneficial effect on biofilm growth – ST:STUDY_SUMMARY with significantly fewer dead bacteria present by biomass and surface area of ST:STUDY_SUMMARY the biofilms (p <0.05). Where arginine and glucose significantly increased and ST:STUDY_SUMMARY decreased pH, respectively, the pH of proline supplemented biofilms remained ST:STUDY_SUMMARY neutral at pH 7.3-7.5. SDS-polyacrylamide gel electrophoresis of the spent ST:STUDY_SUMMARY saliva from proline and arginine supplemented biofilms showed inhibition of ST:STUDY_SUMMARY salivary protein degradation of immature biofilms. NMR analysis of the spent ST:STUDY_SUMMARY saliva revealed that proline supplemented biofilms were metabolically similar to ST:STUDY_SUMMARY unsupplemented biofilms, but these biofilms actively metabolised proline to ST:STUDY_SUMMARY 5-aminopentanoate, butyrate and propionate, and actively utilised glycine. This ST:STUDY_SUMMARY study shows that in a nutrient limited environment, proline has a beneficial ST:STUDY_SUMMARY effect on in vitro oral biofilms grown from a saliva inoculum. ST:INSTITUTE King's College London ST:DEPARTMENT Centre for Host Microbiome Interactions ST:LAST_NAME Cleaver ST:FIRST_NAME Leanne ST:ADDRESS Floor 17, Tower Wing, Guy's Hospital, Great Maze Pond ST:EMAIL leanne.cleaver@kcl.ac.uk ST:PHONE 07464626438 ST:NUM_GROUPS 11 #SUBJECT SU:SUBJECT_TYPE Bacteria SU:SUBJECT_SPECIES Multi-species non-defined biofilm consortium #FACTORS #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS Sterile Saliva sample1 Treatment:Growth Medium RAW_FILE_NAME=sample1.raw SUBJECT_SAMPLE_FACTORS P10 1 D4 sample2 Treatment:Proline 10mM RAW_FILE_NAME=sample2.raw SUBJECT_SAMPLE_FACTORS P10 2 D4 sample3 Treatment:Proline 10mM RAW_FILE_NAME=sample3.raw SUBJECT_SAMPLE_FACTORS P10 3 D4 sample4 Treatment:Proline 10mM RAW_FILE_NAME=sample4.raw SUBJECT_SAMPLE_FACTORS A10 1 D4 sample5 Treatment:Arginine 10mM RAW_FILE_NAME=sample5.raw SUBJECT_SAMPLE_FACTORS A10 2 D4 sample6 Treatment:Arginine 10mM RAW_FILE_NAME=sample6.raw SUBJECT_SAMPLE_FACTORS A10 3 D4 sample7 Treatment:Arginine 10mM RAW_FILE_NAME=sample7.raw SUBJECT_SAMPLE_FACTORS G10 1 D4 sample8 Treatment:Glucose 10mM RAW_FILE_NAME=sample8.raw SUBJECT_SAMPLE_FACTORS G10 2 D4 sample9 Treatment:Glucose 10mM RAW_FILE_NAME=sample9.raw SUBJECT_SAMPLE_FACTORS G10 3 D4 sample10 Treatment:Glucose 10mM RAW_FILE_NAME=sample10.raw SUBJECT_SAMPLE_FACTORS P25 1 D4 sample11 Treatment:Proline 25mM RAW_FILE_NAME=sample11.raw SUBJECT_SAMPLE_FACTORS P25 2 D4 sample12 Treatment:Proline 25mM RAW_FILE_NAME=sample12.raw SUBJECT_SAMPLE_FACTORS P25 3 D4 sample13 Treatment:Proline 25mM RAW_FILE_NAME=sample13.raw SUBJECT_SAMPLE_FACTORS A25 1 D4 sample14 Treatment:Arginine 25mM RAW_FILE_NAME=sample14.raw SUBJECT_SAMPLE_FACTORS A25 2 D4 sample15 Treatment:Arginine 25mM RAW_FILE_NAME=sample15.raw SUBJECT_SAMPLE_FACTORS A25 3 D4 sample16 Treatment:Arginine 25mM RAW_FILE_NAME=sample16.raw SUBJECT_SAMPLE_FACTORS G25 1 D4 sample17 Treatment:Glucose 25mM RAW_FILE_NAME=sample17.raw SUBJECT_SAMPLE_FACTORS G25 2 D4 sample18 Treatment:Glucose 25mM RAW_FILE_NAME=sample18.raw SUBJECT_SAMPLE_FACTORS G25 3 D4 sample19 Treatment:Glucose 25mM RAW_FILE_NAME=sample19.raw SUBJECT_SAMPLE_FACTORS P50 1 D4 sample20 Treatment:Proline 50mM RAW_FILE_NAME=sample20.raw SUBJECT_SAMPLE_FACTORS P50 2 D4 sample21 Treatment:Proline 50mM RAW_FILE_NAME=sample21.raw SUBJECT_SAMPLE_FACTORS P50 3 D4 sample22 Treatment:Proline 50mM RAW_FILE_NAME=sample22.raw SUBJECT_SAMPLE_FACTORS A50 1 D4 sample23 Treatment:Arginine 50mM RAW_FILE_NAME=sample23.raw SUBJECT_SAMPLE_FACTORS A50 2 D4 sample24 Treatment:Arginine 50mM RAW_FILE_NAME=sample24.raw SUBJECT_SAMPLE_FACTORS A50 3 D4 sample25 Treatment:Arginine 50mM RAW_FILE_NAME=sample25.raw SUBJECT_SAMPLE_FACTORS G50 1 D4 sample26 Treatment:Glucose 50mM RAW_FILE_NAME=sample26.raw SUBJECT_SAMPLE_FACTORS G50 2 D4 sample27 Treatment:Glucose 50mM RAW_FILE_NAME=sample27.raw SUBJECT_SAMPLE_FACTORS G50 3 D4 sample28 Treatment:Glucose 50mM RAW_FILE_NAME=sample28.raw SUBJECT_SAMPLE_FACTORS Unsupplemented Control 1 D4 sample29 Treatment:No treatment RAW_FILE_NAME=sample29.raw SUBJECT_SAMPLE_FACTORS Unsupplemented Control 2 D4 sample30 Treatment:No treatment RAW_FILE_NAME=sample30.raw SUBJECT_SAMPLE_FACTORS Unsupplemented Control 3 D4 sample31 Treatment:No treatment RAW_FILE_NAME=sample31.raw SUBJECT_SAMPLE_FACTORS Negative Control 1 D4 sample32 Treatment:No treatment RAW_FILE_NAME=sample32.raw SUBJECT_SAMPLE_FACTORS Negative Control 2 D4 sample33 Treatment:No treatment RAW_FILE_NAME=sample33.raw SUBJECT_SAMPLE_FACTORS Negative Control 3 D4 sample34 Treatment:No treatment RAW_FILE_NAME=sample34.raw SUBJECT_SAMPLE_FACTORS P10 1 D7 sample35 Treatment:Proline 10mM RAW_FILE_NAME=sample35.raw SUBJECT_SAMPLE_FACTORS P10 2 D7 sample36 Treatment:Proline 10mM RAW_FILE_NAME=sample36.raw SUBJECT_SAMPLE_FACTORS P10 3 D7 sample37 Treatment:Proline 10mM RAW_FILE_NAME=sample37.raw SUBJECT_SAMPLE_FACTORS A10 1 D7 sample38 Treatment:Arginine 10mM RAW_FILE_NAME=sample38.raw SUBJECT_SAMPLE_FACTORS A10 2 D7 sample39 Treatment:Arginine 10mM RAW_FILE_NAME=sample39.raw SUBJECT_SAMPLE_FACTORS A10 3 D7 sample40 Treatment:Arginine 10mM RAW_FILE_NAME=sample40.raw SUBJECT_SAMPLE_FACTORS G10 1 D7 sample41 Treatment:Glucose 10mM RAW_FILE_NAME=sample41.raw SUBJECT_SAMPLE_FACTORS G10 2 D7 sample42 Treatment:Glucose 10mM RAW_FILE_NAME=sample42.raw SUBJECT_SAMPLE_FACTORS G10 3 D7 sample43 Treatment:Glucose 10mM RAW_FILE_NAME=sample43.raw SUBJECT_SAMPLE_FACTORS P25 1 D7 sample44 Treatment:Proline 25mM RAW_FILE_NAME=sample44.raw SUBJECT_SAMPLE_FACTORS P25 2 D7 sample45 Treatment:Proline 25mM RAW_FILE_NAME=sample45.raw SUBJECT_SAMPLE_FACTORS P25 3 D7 sample46 Treatment:Proline 25mM RAW_FILE_NAME=sample46.raw SUBJECT_SAMPLE_FACTORS A25 1 D7 sample47 Treatment:Arginine 25mM RAW_FILE_NAME=sample47.raw SUBJECT_SAMPLE_FACTORS A25 2 D7 sample48 Treatment:Arginine 25mM RAW_FILE_NAME=sample48.raw SUBJECT_SAMPLE_FACTORS A25 3 D7 sample49 Treatment:Arginine 25mM RAW_FILE_NAME=sample49.raw SUBJECT_SAMPLE_FACTORS G25 1 D7 sample50 Treatment:Glucose 25mM RAW_FILE_NAME=sample50.raw SUBJECT_SAMPLE_FACTORS G25 2 D7 sample51 Treatment:Glucose 25mM RAW_FILE_NAME=sample51.raw SUBJECT_SAMPLE_FACTORS G25 3 D7 sample52 Treatment:Glucose 25mM RAW_FILE_NAME=sample52.raw SUBJECT_SAMPLE_FACTORS P50 1 D7 sample53 Treatment:Proline 50mM RAW_FILE_NAME=sample53.raw SUBJECT_SAMPLE_FACTORS P50 2 D7 sample54 Treatment:Proline 50mM RAW_FILE_NAME=sample54.raw SUBJECT_SAMPLE_FACTORS P50 3 D7 sample55 Treatment:Proline 50mM RAW_FILE_NAME=sample55.raw SUBJECT_SAMPLE_FACTORS A50 1 D7 sample56 Treatment:Arginine 50mM RAW_FILE_NAME=sample56.raw SUBJECT_SAMPLE_FACTORS A50 2 D7 sample57 Treatment:Arginine 50mM RAW_FILE_NAME=sample57.raw SUBJECT_SAMPLE_FACTORS A50 3 D7 sample58 Treatment:Arginine 50mM RAW_FILE_NAME=sample58.raw SUBJECT_SAMPLE_FACTORS G50 1 D7 sample59 Treatment:Glucose 50mM RAW_FILE_NAME=sample59.raw SUBJECT_SAMPLE_FACTORS G50 2 D7 sample60 Treatment:Glucose 50mM RAW_FILE_NAME=sample60.raw SUBJECT_SAMPLE_FACTORS G50 3 D7 sample61 Treatment:Glucose 50mM RAW_FILE_NAME=sample61.raw SUBJECT_SAMPLE_FACTORS Unsupplemented Control 1 D7 sample62 Treatment:No treatment RAW_FILE_NAME=sample62.raw SUBJECT_SAMPLE_FACTORS Unsupplemented Control 2 D7 sample63 Treatment:No treatment RAW_FILE_NAME=sample63.raw SUBJECT_SAMPLE_FACTORS Unsupplemented Control 3 D7 sample64 Treatment:No treatment RAW_FILE_NAME=sample64.raw SUBJECT_SAMPLE_FACTORS Negative Control 1 D7 sample65 Treatment:No treatment RAW_FILE_NAME=sample65.raw SUBJECT_SAMPLE_FACTORS Negative Control 2 D7 sample66 Treatment:No treatment RAW_FILE_NAME=sample66.raw SUBJECT_SAMPLE_FACTORS Negative Control 3 D7 sample67 Treatment:No treatment RAW_FILE_NAME=sample67.raw #COLLECTION CO:COLLECTION_SUMMARY Saliva was processed and biofilms were grown using a previously published CO:COLLECTION_SUMMARY methodology with modifications (Cleaver, Moazzez and Carpenter, 2019). Saliva CO:COLLECTION_SUMMARY samples were centrifuged (5000g for 5 mins), and the supernatant was collected, CO:COLLECTION_SUMMARY pooled, boiled in sealed tubes for 20 minutes to sterilise, and left to cool to CO:COLLECTION_SUMMARY room temperature; hereafter referred to as sterile saliva. The loose pellet from CO:COLLECTION_SUMMARY the spun saliva samples was pooled and combined with enough sterile saliva to CO:COLLECTION_SUMMARY facilitate inoculation of the hydroxyapatite (HA) discs. Samples were vortexed CO:COLLECTION_SUMMARY vigorously to resuspend the pellet; this constituted the pooled saliva inoculum. CO:COLLECTION_SUMMARY The remaining sterile saliva was split into 11 aliquots. D-proline, L-arginine, CO:COLLECTION_SUMMARY and glucose were added to the aliquots to achieve concentrations of 10mM, 25mM CO:COLLECTION_SUMMARY and 50mM respectively for each (based on concentrations of proline obtained from CO:COLLECTION_SUMMARY previous publication (6)). One aliquot was also supplemented with 10mM carbon 13 CO:COLLECTION_SUMMARY labelled (C13) D-proline. Two aliquots were left unsupplemented to act as a CO:COLLECTION_SUMMARY positive and negative control. Six HA discs in a sterile microtitre plate were CO:COLLECTION_SUMMARY inoculated per treatment condition with 1 ml pooled saliva inoculum (negative CO:COLLECTION_SUMMARY control discs were inoculated with sterile saliva only to check for sterility) CO:COLLECTION_SUMMARY and incubated for 24 hours aerobically in a 40 L aerobic incubator (GenLab Ltd, CO:COLLECTION_SUMMARY UK) at 37°C. After this time the discs were washed three times with sterile CO:COLLECTION_SUMMARY phosphate buffered saline (PBS) and 1 ml of supplemented/unsupplemented saliva CO:COLLECTION_SUMMARY per condition was added to the discs. The discs were then incubated CO:COLLECTION_SUMMARY anaerobically in a 3.5 litre anaerobic jar with an Anaerogen anaerobic generator CO:COLLECTION_SUMMARY pack (Oxoid, UK) at 37°C for 72 hours. After this spent saliva from the CO:COLLECTION_SUMMARY biofilms was removed and stored at -20°C for further analysis, discs were CO:COLLECTION_SUMMARY washed three times with PBS, and refreshed with supplemented/unsupplemented CO:COLLECTION_SUMMARY saliva and further incubated anaerobically at 37°C for 72 hours. After this CO:COLLECTION_SUMMARY final incubation the spent saliva was removed and stored at -20°C for further CO:COLLECTION_SUMMARY analysis. CO:SAMPLE_TYPE Bacterial cells #TREATMENT TR:TREATMENT_SUMMARY Sterile saliva was split into 11 aliquots. D-proline, L-arginine, and glucose TR:TREATMENT_SUMMARY were added to the aliquots to achieve concentrations of 10mM, 25mM and 50mM TR:TREATMENT_SUMMARY respectively for each (based on concentrations of proline obtained from previous TR:TREATMENT_SUMMARY publication (6)). One aliquot was also supplemented with 10mM carbon 13 labelled TR:TREATMENT_SUMMARY (C13) D-proline. Two aliquots were left unsupplemented to act as a positive and TR:TREATMENT_SUMMARY negative control. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Samples were processed for NMR using a previously published method (6). Briefly, SP:SAMPLEPREP_SUMMARY centrifuged spent saliva supernatant and the sterile saliva sample were each SP:SAMPLEPREP_SUMMARY mixed with TSP buffer in a 5 mm NMR tube (Bruker, Germany). The tubes were SP:SAMPLEPREP_SUMMARY sealed and analysed at the Biomolecular Spectroscopy Centre, King’s College SP:SAMPLEPREP_SUMMARY London, UK on a 600 MHz spectrometer (Bruker) for 1H 1D-NMR and 1H-C13 1D- and SP:SAMPLEPREP_SUMMARY 2D-NMR. The concentration of metabolites relative to the sterile saliva baseline SP:SAMPLEPREP_SUMMARY were collected using Chenomix NMR Suite version 8.5 (Chenomix Ltd, Canada). The SP:SAMPLEPREP_SUMMARY 2D C13 spectra were analysed using TopSpin version 3.6.2 (Bruker) and COLMAR SP:SAMPLEPREP_SUMMARY (30). #ANALYSIS AN:ANALYSIS_TYPE NMR #NMR NM:INSTRUMENT_NAME Bruker NM:INSTRUMENT_TYPE FT-NMR NM:NMR_EXPERIMENT_TYPE 1D-1H NM:SPECTROMETER_FREQUENCY 600Hz #NMR_METABOLITE_DATA NMR_METABOLITE_DATA:UNITS mM NMR_METABOLITE_DATA_START Samples sample1 sample2 sample3 sample4 sample5 sample6 sample7 sample8 sample9 sample10 sample11 sample12 sample13 sample14 sample15 sample16 sample17 sample18 sample19 sample20 sample21 sample22 sample23 sample24 sample25 sample26 sample27 sample28 sample29 sample30 sample31 sample32 sample33 sample34 sample35 sample36 sample37 sample38 sample39 sample40 sample41 sample42 sample43 sample44 sample45 sample46 sample47 sample48 sample49 sample50 sample51 sample52 sample53 sample54 sample55 sample56 sample57 sample58 sample59 sample60 sample61 sample62 sample63 sample64 sample65 sample66 sample67 Factors Treatment:Growth Medium Treatment:Proline 10mM Treatment:Proline 10mM Treatment:Proline 10mM Treatment:Arginine 10mM Treatment:Arginine 10mM Treatment:Arginine 10mM Treatment:Glucose 10mM Treatment:Glucose 10mM Treatment:Glucose 10mM Treatment:Proline 25mM Treatment:Proline 25mM Treatment:Proline 25mM Treatment:Arginine 25mM Treatment:Arginine 25mM Treatment:Arginine 25mM Treatment:Glucose 25mM Treatment:Glucose 25mM Treatment:Glucose 25mM Treatment:Proline 50mM Treatment:Proline 50mM Treatment:Proline 50mM Treatment:Arginine 50mM Treatment:Arginine 50mM Treatment:Arginine 50mM Treatment:Glucose 50mM Treatment:Glucose 50mM Treatment:Glucose 50mM Treatment:No treatment Treatment:No treatment Treatment:No treatment Treatment:No treatment Treatment:No treatment Treatment:No treatment Treatment:Proline 10mM Treatment:Proline 10mM Treatment:Proline 10mM Treatment:Arginine 10mM Treatment:Arginine 10mM Treatment:Arginine 10mM Treatment:Glucose 10mM Treatment:Glucose 10mM Treatment:Glucose 10mM Treatment:Proline 25mM Treatment:Proline 25mM Treatment:Proline 25mM Treatment:Arginine 25mM Treatment:Arginine 25mM Treatment:Arginine 25mM Treatment:Glucose 25mM Treatment:Glucose 25mM Treatment:Glucose 25mM Treatment:Proline 50mM Treatment:Proline 50mM Treatment:Proline 50mM Treatment:Arginine 50mM Treatment:Arginine 50mM Treatment:Arginine 50mM Treatment:Glucose 50mM Treatment:Glucose 50mM Treatment:Glucose 50mM Treatment:No treatment Treatment:No treatment Treatment:No treatment Treatment:No treatment Treatment:No treatment Treatment:No treatment 5-Aminopentanoic acid 0 1.5654 1.2133 1.497 0.9911 0.6478 0.6567 0.8604 0.607 0.7329 1.4586 1.082 1.3326 0.1244 0.1105 0.0909 0 0.1831 0.171 1.3569 1.4923 1.2555 0.0926 0.1034 0.1 0 0.141 0 1.2992 1.3429 1.0774 0.1125 0.097 0.0942 0.7139 0.479 0.8383 1.244 1.0811 1.2903 0.3094 0.4528 0.4444 0.8435 0.993 0.7779 0.136 0.0926 0.0944 0.0966 0.0894 0.3468 0.9219 1.0341 0.9383 0.0831 0.1027 0.0821 0.081 0 0.104 2.3716 2.4505 1.7254 0.0909 0 0.1122 Propionic acid 0.2998 0.4734 0.4305 0.4649 0.3957 0.4064 0.4493 0.2783 0.5404 0.7805 0.514 0.3812 0.4689 0.3322 0.3114 0.2631 0.1716 0.1792 0.1736 0.5653 0.5713 0.6098 0.2473 0.2697 0.2945 0.1411 0.1638 0.1614 0.3945 0.3777 0.365 0.2722 0.249 0.218 0.7021 0.5379 0.4774 0.2986 0.3133 0.3461 0.2146 1.4394 1.2878 0.6322 0.7363 0.9528 0.3631 0.2198 0.2398 0.1895 0.1785 0.15 0.6691 0.5361 0.6499 0.2283 0.2674 0.2105 0.1843 0.1834 0.2223 0.3926 0.4303 0.2997 0.2271 0.13 0.2255 Butyric acid 0 0.8958 0.7544 0.8345 0.5692 0.5254 0.6559 0.0595 0.1175 0.222 0.827 0.7416 0.7288 0.0752 0.0688 0.0505 0.0389 0.0428 0.0361 0.9427 0.9134 0.8441 0.0509 0.0558 0.0613 0.0256 0.0283 0.0284 0.496 0.1439 0.324 0.0389 0.037 0.0338 1.2285 1.1275 0.8096 0.4204 0.3626 0.366 0.0482 0.3053 0.0518 0.9916 1.0682 1.1235 0.0777 0.0442 0.0495 0.0359 0.032 0.0358 0.9988 0.9367 0.8815 0.0452 0.0563 0.0437 0.0389 0.0311 0.0375 0.9169 0.8141 0.6562 0.0333 0 0.0345 Formic acid 0.0985 2.6695 1.6217 0.5505 1.5973 1.4291 1.3268 4.5712 3.4475 4.7489 1.7439 1.0552 1.0519 1.19 1.2907 0.9776 0.4044 0.7437 0.8219 1.3699 1.1744 0.68 0.8192 0.7415 1.1531 0.2516 0.4002 0.2569 0.1132 0.7764 0.9212 0.0845 0.0628 0.0783 0.244 0.0019 0 0.2049 0.512 1.2597 1.7747 4.0336 3.1813 0.1744 0.0801 0.1304 0.8626 0.5828 1.0689 0.1239 0.055 0.2983 0.0068 0.0451 0.0335 0.611 0.6522 0.5172 0.0385 0.5788 0.0732 0.7838 0.5065 0.6417 0 0.0888 0.0784 Ethanol 0 0.2079 0.1804 0.1983 0.2056 0.2583 0.2702 0.4515 0.3359 0.4398 0.133 0.1103 0.125 0.5247 0.4738 0.402 0.4251 0.4126 0.4171 0.1532 0.1484 0.1535 0.3958 0.4094 0.4352 0.2651 0.3242 0.281 0.2261 0.2119 0.235 0.4181 0.3854 0.3166 0.7769 0.7835 0.6233 0.7268 0.7109 0.7454 0.8141 0.794 0.8607 0.5354 0.663 0.6988 1.2266 0.7867 0.8576 0.9935 0.9409 1.0338 0.4825 0.4774 0.5208 0.8509 0.862 0.757 0.8758 0.8921 1.0411 1.0385 1.0762 1.0609 1.2718 0.897 1.1264 Acetic acid 2.1818 9.021 7.9856 8.7387 6.9626 7.2142 8.1821 7.4442 6.1854 7.9605 8.6239 7.3983 7.7907 4.492 4.0641 3.4935 2.2735 2.5761 2.6142 9.5797 9.3113 8.8818 3.2345 3.56 3.9599 1.6903 2.2496 1.7313 6.377 6.0199 6.9844 2.0251 1.8528 1.6413 12.1875 10.6843 9.2826 5.9131 6.332 6.9559 4.1082 7.6525 6.5859 11.194 11.8059 11.9926 4.1478 2.6911 3.3318 1.5863 1.4287 1.7548 9.9783 10.437 9.5848 2.6612 3.2183 2.5034 1.4363 1.8431 1.6418 10.0832 10.0404 8.2539 1.7943 1.8702 1.7765 Phenylalanine 0 0.0589 0.0534 0.0573 0.0391 0.0451 0.0527 0.0526 0.0379 0.0446 0.062 0.0511 0.0522 0.0485 0.0419 0.0336 0.0278 0.0269 0.0249 0.0542 0.0594 0.0559 0.0363 0.035 0.0391 0.0251 0.0286 0.0261 0 0 0.0529 0 0 0 0.0604 0.0642 0.0461 0.0324 0.0334 0.0365 0.0343 0.0436 0.0336 0.0531 0.0641 0.0729 0.0438 0 0.0304 0.0331 0.0325 0.0331 0.0546 0.0521 0.0473 0.0258 0.0329 0.0268 0.0311 0.0304 0.0065 0.0476 0.0556 0.0318 0 0 0 Alanine 0.019 0 0 0 0 0.0759 0.1358 0.1603 0.1134 0.1297 0 0 0 0.2194 0.1774 0.1454 0.0804 0.0727 0.0656 0 0 0 0.135 0.1469 0.156 0 0.0817 0 0 0 0 0.016 0.0146 0.0133 0 0 0 0.0233 0 0.0316 0.1045 0.1204 0.0694 0 0 0 0.177 0.1031 0.1423 0.0542 0 0.0767 0 0 0 0.0946 0.1688 0.114 0 0 0.0098 0 0 0 0.0129 0 0.0146 Lactic acid 0.1683 0.0048 0.0039 0.0086 0.0073 0.0267 0.0281 8.6563 5.8085 7.1043 0.0064 0.0037 0.004 0.2784 0.1667 0.1613 13.3291 13.5382 13.1396 0.006 0.0039 0.004 0.2619 0.2844 0.1352 11.0059 13.6119 13.4516 0.006 0.0188 0.0064 0.1465 0.1356 0.1201 0.0047 0.0037 0.004 0.0155 0.0082 0.0164 10.2342 7.9777 7.8013 0.0074 0.0044 0.0046 0.4477 0.2255 0.3623 15.1106 13.1143 14.6809 0.0126 0 0 0.2508 0.2595 0.1795 10.4028 13.0744 0.1516 0.0082 0.0057 0.0103 0.124 0.1184 0.1246 Putrescine 0 0.2748 0.2358 0.2553 3.3936 3.8634 3.7586 0 0 0 0.2622 0.2306 0.2518 5.7624 4.7715 4.4849 0 0 0 0.3011 0.29 0.2777 3.5543 3.7516 4.6764 0 0 0 0.215 0.2405 0.2464 0 0 0 0.3457 0.332 0.2734 2.6905 2.8036 2.4468 0 0 0 0.3223 0.3325 0.4096 5.3743 3.8358 3.3974 0 0 0 0.2659 0.28 0.2412 2.9041 5.5422 4.1946 0 0 0 0.2824 0.29 0.2145 0 0 0 Methanol 0.0242 0.1019 0.0864 0.0926 0.0698 0.0872 0.0872 0.0689 0.0516 0.0684 0.6982 0.6049 0.654 0.0951 0.0886 0.0748 0.0727 0.0687 0.0707 1.3552 1.2888 1.2279 0.0731 0.0725 0.0783 0.0693 0.0794 0.0729 0.0835 0.0838 0.087 0.0883 0.0892 0.079 0.1657 0.1685 0.1395 0.1322 0.1278 0.1368 0.1062 0.1051 0.1113 0.8419 0.1888 1.0258 0.1652 0.1044 0.111 0.1209 0.1144 0.1253 1.2542 0.1902 0.227 0.1127 0.1176 0.1011 0.1169 0.1179 0.1339 0.15 0.1519 0.1472 0.1636 0.1327 0.1426 Proline 0 6.6555 5.58 6.0444 1.1481 1.3558 1.3206 0 0 0 14.087 12.0003 13.1289 0.6823 0.6464 0.575 0.3187 0.3176 0 25.2884 25.4488 24.0924 0.4117 0.4181 0.5329 0 0 0 0.1843 0.4324 0.8055 0 0 0 9.1519 8.663 5.3857 0 0.4241 0 0.5385 0.3228 0.5624 17.1239 18.2361 19.778 0.5749 0.371 0.4342 0 0.2184 0.2883 24.8854 25.8482 23.5506 0.3099 0.3855 0.3031 0 0 0 0 0.1029 0 0 0 0 Succinic acid 0.0248 0.376 0.3208 0.4332 0.163 0.2138 0.2288 0.1093 0.0293 0.0308 0.3504 0.2983 0.3699 0.1198 0.1162 0.1043 0.0659 0.0752 0.0835 0.4123 0.3888 0.3608 0.0696 0.0748 0.1177 0.043 0.0619 0.04 0.159 0.2987 0.3788 0.0212 0 0.0174 0.4413 0.4566 0.3534 0.1152 0.1411 0.0846 0.0716 0 0 0.4591 0.4226 0.4026 0.0672 0.0417 0.0385 0.0375 0.0362 0.0359 0.377 0.2932 0.2614 0.0254 0.0307 0.0135 0.0265 0.0253 0.0167 0.4009 0.532 0.4034 0 0.0182 0.0179 Glycine 0.0778 0.1094 0.0094 0.0081 0.0101 0 0.0082 1.1934 0.8007 1.0387 0.0077 0.0068 0.0064 0.4012 0.35 0.3327 0.203 0.2017 0.1975 0.0014 0 0 0.2538 0.2634 0.3041 0.176 0.198 0.1708 0.6765 0.8657 0.8305 0.0714 0.0644 0.0578 0.0053 0.0053 0.0072 0.1564 0.0074 0.3186 0.7397 0.9807 0.9317 0.0082 0.0022 0 0.3648 0.1991 0.0131 0.1417 0.1375 0.4763 0 0 0 0.1921 0.2459 0.1925 0.1289 0.1465 0.055 0.66 0.0072 0.0072 0.0588 0.07 0.0657 Glutamic acid 0 0 0 0 0 0 0 0.7505 0.5168 0.5104 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.5116 0.2741 0 0 0 0 0 0 0 0 0 0.7853 0.3403 0.6926 0 0 0 0 0 0 0 0 0.5707 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Valine 0 0.1013 0.0751 0.0787 0.047 0.0547 0.0611 0.0542 0.039 0.0482 0.0805 0.0662 0.0722 0.0362 0.0367 0.0299 0.0264 0.0268 0.0236 0.0898 0.0874 0.0892 0.0262 0.03 0.0375 0.025 0.0305 0.0287 0.025 0.0573 0.0666 0 0 0 0.1001 0.0944 0.0697 0.0184 0.0262 0.0255 0.0454 0.0441 0.0465 0.0883 0.0922 0.1014 0.0197 0.0139 0.0186 0.0276 0.0266 0.0461 0.0775 0.0705 0.0676 0.0144 0.0159 0.0166 0.0176 0.0527 0.0017 0.0688 0.0767 0.0571 0 0 0 Glucose 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 4.8326 5.3765 4.8441 0 0 0 0 0 0 11.6622 13.972 11.4724 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 6.2758 6.0624 4.4247 0 0 0 0 0 0 13.6535 13.4713 19.3396 0 0 0 0 0 0 Tyrosine 0.0249 0 0 0 0 0 0 0 0.0607 0.0356 0 0 0 0.0633 0.0494 0.04 0.0464 0.0515 0.0421 0 0 0 0.0445 0.0379 0.0479 0.0435 0.0467 0.0435 0 0.0281 0.0098 0 0 0.0136 0 0 0 0 0 0 0.0737 0 0.052 0 0 0 0.0719 0.0439 0.0504 0.049 0.0468 0.0496 0 0 0 0.0388 0.0516 0.0413 0.0491 0.0446 0.0134 0 0 0 0 0 0.0156 Arginine 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2.3074 0.0996 3.2188 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.2259 0 0.0664 0 0 0 0 0 0 0.0963 0 0.7123 0 0 0 0 0 0 0 0 0 NMR_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Sterile Saliva P10 1 D4 P10 2 D4 P10 3 D4 A10 1 D4 A10 2 D4 A10 3 D4 G10 1 D4 G10 2 D4 G10 3 D4 P25 1 D4 P25 2 D4 P25 3 D4 A25 1 D4 A25 2 D4 A25 3 D4 G25 1 D4 G25 2 D4 G25 3 D4 P50 1 D4 P50 2 D4 P50 3 D4 A50 1 D4 A50 2 D4 A50 3 D4 G50 1 D4 G50 2 D4 G50 3 D4 Unsupplemented Control 1 D4 Unsupplemented Control 2 D4 Unsupplemented Control 3 D4 Negative Control 1 D4 Negative Control 2 D4 Negative Control 3 D4 P10 1 D7 P10 2 D7 P10 3 D7 A10 1 D7 A10 2 D7 A10 3 D7 G10 1 D7 G10 2 D7 G10 3 D7 P25 1 D7 P25 2 D7 P25 3 D7 A25 1 D7 A25 2 D7 A25 3 D7 G25 1 D7 G25 2 D7 G25 3 D7 P50 1 D7 P50 2 D7 P50 3 D7 A50 1 D7 A50 2 D7 A50 3 D7 G50 1 D7 G50 2 D7 G50 3 D7 Unsupplemented Control 1 D7 Unsupplemented Control 2 D7 Unsupplemented Control 3 D7 Negative Control 1 D7 Negative Control 2 D7 Negative Control 3 D7 5-Aminopentanoic acid 0 1.5654 1.2133 1.497 0.9911 0.6478 0.6567 0.8604 0.607 0.7329 1.4586 1.082 1.3326 0.1244 0.1105 0.0909 0 0.1831 0.171 1.3569 1.4923 1.2555 0.0926 0.1034 0.1 0 0.141 0 1.2992 1.3429 1.0774 0.1125 0.097 0.0942 0.7139 0.479 0.8383 1.244 1.0811 1.2903 0.3094 0.4528 0.4444 0.8435 0.993 0.7779 0.136 0.0926 0.0944 0.0966 0.0894 0.3468 0.9219 1.0341 0.9383 0.0831 0.1027 0.0821 0.081 0 0.104 2.3716 2.4505 1.7254 0.0909 0 0.1122 Propionic acid 0.2998 0.4734 0.4305 0.4649 0.3957 0.4064 0.4493 0.2783 0.5404 0.7805 0.514 0.3812 0.4689 0.3322 0.3114 0.2631 0.1716 0.1792 0.1736 0.5653 0.5713 0.6098 0.2473 0.2697 0.2945 0.1411 0.1638 0.1614 0.3945 0.3777 0.365 0.2722 0.249 0.218 0.7021 0.5379 0.4774 0.2986 0.3133 0.3461 0.2146 1.4394 1.2878 0.6322 0.7363 0.9528 0.3631 0.2198 0.2398 0.1895 0.1785 0.15 0.6691 0.5361 0.6499 0.2283 0.2674 0.2105 0.1843 0.1834 0.2223 0.3926 0.4303 0.2997 0.2271 0.13 0.2255 Butyric acid 0 0.8958 0.7544 0.8345 0.5692 0.5254 0.6559 0.0595 0.1175 0.222 0.827 0.7416 0.7288 0.0752 0.0688 0.0505 0.0389 0.0428 0.0361 0.9427 0.9134 0.8441 0.0509 0.0558 0.0613 0.0256 0.0283 0.0284 0.496 0.1439 0.324 0.0389 0.037 0.0338 1.2285 1.1275 0.8096 0.4204 0.3626 0.366 0.0482 0.3053 0.0518 0.9916 1.0682 1.1235 0.0777 0.0442 0.0495 0.0359 0.032 0.0358 0.9988 0.9367 0.8815 0.0452 0.0563 0.0437 0.0389 0.0311 0.0375 0.9169 0.8141 0.6562 0.0333 0 0.0345 Formic acid 0.0985 2.6695 1.6217 0.5505 1.5973 1.4291 1.3268 4.5712 3.4475 4.7489 1.7439 1.0552 1.0519 1.19 1.2907 0.9776 0.4044 0.7437 0.8219 1.3699 1.1744 0.68 0.8192 0.7415 1.1531 0.2516 0.4002 0.2569 0.1132 0.7764 0.9212 0.0845 0.0628 0.0783 0.244 0.0019 0 0.2049 0.512 1.2597 1.7747 4.0336 3.1813 0.1744 0.0801 0.1304 0.8626 0.5828 1.0689 0.1239 0.055 0.2983 0.0068 0.0451 0.0335 0.611 0.6522 0.5172 0.0385 0.5788 0.0732 0.7838 0.5065 0.6417 0 0.0888 0.0784 Ethanol 0 0.2079 0.1804 0.1983 0.2056 0.2583 0.2702 0.4515 0.3359 0.4398 0.133 0.1103 0.125 0.5247 0.4738 0.402 0.4251 0.4126 0.4171 0.1532 0.1484 0.1535 0.3958 0.4094 0.4352 0.2651 0.3242 0.281 0.2261 0.2119 0.235 0.4181 0.3854 0.3166 0.7769 0.7835 0.6233 0.7268 0.7109 0.7454 0.8141 0.794 0.8607 0.5354 0.663 0.6988 1.2266 0.7867 0.8576 0.9935 0.9409 1.0338 0.4825 0.4774 0.5208 0.8509 0.862 0.757 0.8758 0.8921 1.0411 1.0385 1.0762 1.0609 1.2718 0.897 1.1264 Acetic acid 2.1818 9.021 7.9856 8.7387 6.9626 7.2142 8.1821 7.4442 6.1854 7.9605 8.6239 7.3983 7.7907 4.492 4.0641 3.4935 2.2735 2.5761 2.6142 9.5797 9.3113 8.8818 3.2345 3.56 3.9599 1.6903 2.2496 1.7313 6.377 6.0199 6.9844 2.0251 1.8528 1.6413 12.1875 10.6843 9.2826 5.9131 6.332 6.9559 4.1082 7.6525 6.5859 11.194 11.8059 11.9926 4.1478 2.6911 3.3318 1.5863 1.4287 1.7548 9.9783 10.437 9.5848 2.6612 3.2183 2.5034 1.4363 1.8431 1.6418 10.0832 10.0404 8.2539 1.7943 1.8702 1.7765 Phenylalanine 0 0.0589 0.0534 0.0573 0.0391 0.0451 0.0527 0.0526 0.0379 0.0446 0.062 0.0511 0.0522 0.0485 0.0419 0.0336 0.0278 0.0269 0.0249 0.0542 0.0594 0.0559 0.0363 0.035 0.0391 0.0251 0.0286 0.0261 0 0 0.0529 0 0 0 0.0604 0.0642 0.0461 0.0324 0.0334 0.0365 0.0343 0.0436 0.0336 0.0531 0.0641 0.0729 0.0438 0 0.0304 0.0331 0.0325 0.0331 0.0546 0.0521 0.0473 0.0258 0.0329 0.0268 0.0311 0.0304 0.0065 0.0476 0.0556 0.0318 0 0 0 Alanine 0.019 0 0 0 0 0.0759 0.1358 0.1603 0.1134 0.1297 0 0 0 0.2194 0.1774 0.1454 0.0804 0.0727 0.0656 0 0 0 0.135 0.1469 0.156 0 0.0817 0 0 0 0 0.016 0.0146 0.0133 0 0 0 0.0233 0 0.0316 0.1045 0.1204 0.0694 0 0 0 0.177 0.1031 0.1423 0.0542 0 0.0767 0 0 0 0.0946 0.1688 0.114 0 0 0.0098 0 0 0 0.0129 0 0.0146 Lactic acid 0.1683 0.0048 0.0039 0.0086 0.0073 0.0267 0.0281 8.6563 5.8085 7.1043 0.0064 0.0037 0.004 0.2784 0.1667 0.1613 13.3291 13.5382 13.1396 0.006 0.0039 0.004 0.2619 0.2844 0.1352 11.0059 13.6119 13.4516 0.006 0.0188 0.0064 0.1465 0.1356 0.1201 0.0047 0.0037 0.004 0.0155 0.0082 0.0164 10.2342 7.9777 7.8013 0.0074 0.0044 0.0046 0.4477 0.2255 0.3623 15.1106 13.1143 14.6809 0.0126 0 0 0.2508 0.2595 0.1795 10.4028 13.0744 0.1516 0.0082 0.0057 0.0103 0.124 0.1184 0.1246 Putrescine 0 0.2748 0.2358 0.2553 3.3936 3.8634 3.7586 0 0 0 0.2622 0.2306 0.2518 5.7624 4.7715 4.4849 0 0 0 0.3011 0.29 0.2777 3.5543 3.7516 4.6764 0 0 0 0.215 0.2405 0.2464 0 0 0 0.3457 0.332 0.2734 2.6905 2.8036 2.4468 0 0 0 0.3223 0.3325 0.4096 5.3743 3.8358 3.3974 0 0 0 0.2659 0.28 0.2412 2.9041 5.5422 4.1946 0 0 0 0.2824 0.29 0.2145 0 0 0 Methanol 0.0242 0.1019 0.0864 0.0926 0.0698 0.0872 0.0872 0.0689 0.0516 0.0684 0.6982 0.6049 0.654 0.0951 0.0886 0.0748 0.0727 0.0687 0.0707 1.3552 1.2888 1.2279 0.0731 0.0725 0.0783 0.0693 0.0794 0.0729 0.0835 0.0838 0.087 0.0883 0.0892 0.079 0.1657 0.1685 0.1395 0.1322 0.1278 0.1368 0.1062 0.1051 0.1113 0.8419 0.1888 1.0258 0.1652 0.1044 0.111 0.1209 0.1144 0.1253 1.2542 0.1902 0.227 0.1127 0.1176 0.1011 0.1169 0.1179 0.1339 0.15 0.1519 0.1472 0.1636 0.1327 0.1426 Proline 0 6.6555 5.58 6.0444 1.1481 1.3558 1.3206 0 0 0 14.087 12.0003 13.1289 0.6823 0.6464 0.575 0.3187 0.3176 0 25.2884 25.4488 24.0924 0.4117 0.4181 0.5329 0 0 0 0.1843 0.4324 0.8055 0 0 0 9.1519 8.663 5.3857 0 0.4241 0 0.5385 0.3228 0.5624 17.1239 18.2361 19.778 0.5749 0.371 0.4342 0 0.2184 0.2883 24.8854 25.8482 23.5506 0.3099 0.3855 0.3031 0 0 0 0 0.1029 0 0 0 0 Succinic acid 0.0248 0.376 0.3208 0.4332 0.163 0.2138 0.2288 0.1093 0.0293 0.0308 0.3504 0.2983 0.3699 0.1198 0.1162 0.1043 0.0659 0.0752 0.0835 0.4123 0.3888 0.3608 0.0696 0.0748 0.1177 0.043 0.0619 0.04 0.159 0.2987 0.3788 0.0212 0 0.0174 0.4413 0.4566 0.3534 0.1152 0.1411 0.0846 0.0716 0 0 0.4591 0.4226 0.4026 0.0672 0.0417 0.0385 0.0375 0.0362 0.0359 0.377 0.2932 0.2614 0.0254 0.0307 0.0135 0.0265 0.0253 0.0167 0.4009 0.532 0.4034 0 0.0182 0.0179 Glycine 0.0778 0.1094 0.0094 0.0081 0.0101 0 0.0082 1.1934 0.8007 1.0387 0.0077 0.0068 0.0064 0.4012 0.35 0.3327 0.203 0.2017 0.1975 0.0014 0 0 0.2538 0.2634 0.3041 0.176 0.198 0.1708 0.6765 0.8657 0.8305 0.0714 0.0644 0.0578 0.0053 0.0053 0.0072 0.1564 0.0074 0.3186 0.7397 0.9807 0.9317 0.0082 0.0022 0 0.3648 0.1991 0.0131 0.1417 0.1375 0.4763 0 0 0 0.1921 0.2459 0.1925 0.1289 0.1465 0.055 0.66 0.0072 0.0072 0.0588 0.07 0.0657 Glutamic acid 0 0 0 0 0 0 0 0.7505 0.5168 0.5104 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.5116 0.2741 0 0 0 0 0 0 0 0 0 0.7853 0.3403 0.6926 0 0 0 0 0 0 0 0 0.5707 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Valine 0 0.1013 0.0751 0.0787 0.047 0.0547 0.0611 0.0542 0.039 0.0482 0.0805 0.0662 0.0722 0.0362 0.0367 0.0299 0.0264 0.0268 0.0236 0.0898 0.0874 0.0892 0.0262 0.03 0.0375 0.025 0.0305 0.0287 0.025 0.0573 0.0666 0 0 0 0.1001 0.0944 0.0697 0.0184 0.0262 0.0255 0.0454 0.0441 0.0465 0.0883 0.0922 0.1014 0.0197 0.0139 0.0186 0.0276 0.0266 0.0461 0.0775 0.0705 0.0676 0.0144 0.0159 0.0166 0.0176 0.0527 0.0017 0.0688 0.0767 0.0571 0 0 0 Glucose 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 4.8326 5.3765 4.8441 0 0 0 0 0 0 11.6622 13.972 11.4724 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 6.2758 6.0624 4.4247 0 0 0 0 0 0 13.6535 13.4713 19.3396 0 0 0 0 0 0 Tyrosine 0.0249 0 0 0 0 0 0 0 0.0607 0.0356 0 0 0 0.0633 0.0494 0.04 0.0464 0.0515 0.0421 0 0 0 0.0445 0.0379 0.0479 0.0435 0.0467 0.0435 0 0.0281 0.0098 0 0 0.0136 0 0 0 0 0 0 0.0737 0 0.052 0 0 0 0.0719 0.0439 0.0504 0.049 0.0468 0.0496 0 0 0 0.0388 0.0516 0.0413 0.0491 0.0446 0.0134 0 0 0 0 0 0.0156 Arginine 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2.3074 0.0996 3.2188 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.2259 0 0.0664 0 0 0 0 0 0 0.0963 0 0.7123 0 0 0 0 0 0 0 0 0 METABOLITES_END #END