#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