#METABOLOMICS WORKBENCH biswapriya_20180912_075107 DATATRACK_ID:1502 STUDY_ID:ST001053 ANALYSIS_ID:AN001722 PROJECT_ID:PR000706
VERSION             	1
CREATED_ON             	September 17, 2018, 2:29 pm
#PROJECT
PR:PROJECT_TITLE                 	Skin and Blubber Metabolomics
PR:PROJECT_TYPE                  	Multi-institutional Metabolomics
PR:PROJECT_SUMMARY               	The common bottlenose dolphin (Tursiops truncatus) is carnivorous cetacean
PR:PROJECT_SUMMARY               	thriving in marine environment are one of the most common apex predators found
PR:PROJECT_SUMMARY               	in coastal and estuarine ecosystems. Although recent studies have focused on
PR:PROJECT_SUMMARY               	capturing the circulating metabolomes of these organisms, with respect to
PR:PROJECT_SUMMARY               	pollutants and exposures of the marine environment, the skin and blubber are
PR:PROJECT_SUMMARY               	important protective organs that have not been probed. Using 1HNMR based
PR:PROJECT_SUMMARY               	untargeted metabolomics we quantified 51 metabolites belonging to 74 different
PR:PROJECT_SUMMARY               	metabolic pathways in the skin and blubber of bottle nose dolphins (n=5) samples
PR:PROJECT_SUMMARY               	collected in 2017 from the coast of Mexico. Results indicate that the skin and
PR:PROJECT_SUMMARY               	blubber metabolism are quantitatively different. These metabolite abundances
PR:PROJECT_SUMMARY               	could help discriminate the tissue-types using supervised and unsupervised PCA
PR:PROJECT_SUMMARY               	and PLSDA analysis. Heat maps and random forest analysis point to unique
PR:PROJECT_SUMMARY               	metabolites that are important classifiers of the tissue-type. The altered
PR:PROJECT_SUMMARY               	metabolic patterns, mainly linking fatty acid metabolism and ketogenic amino
PR:PROJECT_SUMMARY               	acids, seem to constitute a characteristic of blubber, while the skin showed
PR:PROJECT_SUMMARY               	diverse metabolites involved in gluceoneogenic pathways. 1H NMR spectra allowed
PR:PROJECT_SUMMARY               	the identification of metabolites associated with these organ types, such as
PR:PROJECT_SUMMARY               	pyruvic acid, arginine, ornithine, 2-hydroxybutyric acid, 3-hydroxyisobutyric
PR:PROJECT_SUMMARY               	acid, and acetic acid, as discriminatory and classifying metabolites. These
PR:PROJECT_SUMMARY               	results would lead to further understanding of dolphin skin and blubber
PR:PROJECT_SUMMARY               	metabolism for better efforts in their conservation as well as a measure of
PR:PROJECT_SUMMARY               	marine pollution and ecotoxicology.
PR:INSTITUTE                     	Wake Forest Baptist Medical Center
PR:DEPARTMENT                    	Department of Internal Medicine
PR:LAST_NAME                     	Misra
PR:FIRST_NAME                    	Biswapriya
PR:ADDRESS                       	Medical Center Boulevard NRC Building, G#43, Medical Center Boulevard, Winston
PR:ADDRESS                       	Salem, NC, 27157, USA
PR:EMAIL                         	bbmisraccb@gmail.com
PR:PHONE                         	3522156040
PR:FUNDING_SOURCE                	NA
PR:PROJECT_COMMENTS              	NA
PR:PUBLICATIONS                  	In process
PR:CONTRIBUTORS                  	Biswapriya B. Misra, Ixchel Mariel Ruiz Hernandez, Gloria Ivonne Hernández
PR:CONTRIBUTORS                  	Bolio, Emanuel Hernandez, Colli Dula Reyna Cristina
#STUDY
ST:STUDY_TITLE                   	1H-NMR Analysis of Skin and Blubber of Nose Dolphin Metabolome Reveal the
ST:STUDY_TITLE                   	Functional Metabolomic Dichotomy of The Organs
ST:STUDY_TYPE                    	Dolphin Skin and Blubber
ST:STUDY_SUMMARY                 	The common bottlenose dolphin (Tursiops truncatus) is carnivorous cetacean
ST:STUDY_SUMMARY                 	thriving in marine environment are one of the most common apex predators found
ST:STUDY_SUMMARY                 	in coastal and estuarine ecosystems. Although recent studies have focused on
ST:STUDY_SUMMARY                 	capturing the circulating metabolomes of these organisms, with respect to
ST:STUDY_SUMMARY                 	pollutants and exposures of the marine environment, the skin and blubber are
ST:STUDY_SUMMARY                 	important protective organs that have not been probed. Using 1HNMR based
ST:STUDY_SUMMARY                 	untargeted metabolomics we quantified 51 metabolites belonging to 74 different
ST:STUDY_SUMMARY                 	metabolic pathways in the skin and blubber of bottle nose dolphins (n=5) samples
ST:STUDY_SUMMARY                 	collected in 2017 from the coast of Mexico. Results indicate that the skin and
ST:STUDY_SUMMARY                 	blubber metabolism are quantitatively different. These metabolite abundances
ST:STUDY_SUMMARY                 	could help discriminate the tissue-types using supervised and unsupervised PCA
ST:STUDY_SUMMARY                 	and PLSDA analysis. Heat maps and random forest analysis point to unique
ST:STUDY_SUMMARY                 	metabolites that are important classifiers of the tissue-type. The altered
ST:STUDY_SUMMARY                 	metabolic patterns, mainly linking fatty acid metabolism and ketogenic amino
ST:STUDY_SUMMARY                 	acids, seem to constitute a characteristic of blubber, while the skin showed
ST:STUDY_SUMMARY                 	diverse metabolites involved in gluceoneogenic pathways. 1H NMR spectra allowed
ST:STUDY_SUMMARY                 	the identification of metabolites associated with these organ types, such as
ST:STUDY_SUMMARY                 	pyruvic acid, arginine, ornithine, 2-hydroxybutyric acid, 3-hydroxyisobutyric
ST:STUDY_SUMMARY                 	acid, and acetic acid, as discriminatory and classifying metabolites. These
ST:STUDY_SUMMARY                 	results would lead to further understanding of dolphin skin and blubber
ST:STUDY_SUMMARY                 	metabolism for better efforts in their conservation as well as a measure of
ST:STUDY_SUMMARY                 	marine pollution and ecotoxicology.
ST:INSTITUTE                     	Wake Forest Baptist Medical Center
ST:DEPARTMENT                    	Department of Internal Medicine
ST:LAST_NAME                     	Misra
ST:FIRST_NAME                    	Biswapriya
ST:ADDRESS                       	Medical Center Boulevard NRC Building, G#43, Medical Center Boulevard, Winston
ST:ADDRESS                       	Salem, NC, 27157, USA
ST:EMAIL                         	bbmisraccb@gmail.com
ST:PHONE                         	3522156040
ST:NUM_GROUPS                    	2
ST:TOTAL_SUBJECTS                	4
ST:NUM_MALES                     	3
ST:NUM_FEMALES                   	1
ST:STUDY_COMMENTS                	NA
ST:PUBLICATIONS                  	In process
#SUBJECT
SU:SUBJECT_TYPE                  	Mammal
SU:SUBJECT_SPECIES               	Tursiops truncatus
SU:TAXONOMY_ID                   	9739
SU:GENDER                        	Male and female
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data
SUBJECT_SAMPLE_FACTORS           	-	G7_1	Tissue:Blubber	
SUBJECT_SAMPLE_FACTORS           	-	G7_2	Tissue:Blubber	
SUBJECT_SAMPLE_FACTORS           	-	G7_3	Tissue:Blubber	
SUBJECT_SAMPLE_FACTORS           	-	G10_1	Tissue:Blubber	
SUBJECT_SAMPLE_FACTORS           	-	G10_2	Tissue:Blubber	
SUBJECT_SAMPLE_FACTORS           	-	G10_3	Tissue:Blubber	
SUBJECT_SAMPLE_FACTORS           	-	G14_1	Tissue:Blubber	
SUBJECT_SAMPLE_FACTORS           	-	G14_2	Tissue:Blubber	
SUBJECT_SAMPLE_FACTORS           	-	G14_3	Tissue:Blubber	
SUBJECT_SAMPLE_FACTORS           	-	G15_1	Tissue:Blubber	
SUBJECT_SAMPLE_FACTORS           	-	G15_2	Tissue:Blubber	
SUBJECT_SAMPLE_FACTORS           	-	G15_3	Tissue:Blubber	
SUBJECT_SAMPLE_FACTORS           	-	P7_1	Tissue:Skin	
SUBJECT_SAMPLE_FACTORS           	-	P7_2	Tissue:Skin	
SUBJECT_SAMPLE_FACTORS           	-	P7_3	Tissue:Skin	
SUBJECT_SAMPLE_FACTORS           	-	P10_1	Tissue:Skin	
SUBJECT_SAMPLE_FACTORS           	-	P10_2	Tissue:Skin	
SUBJECT_SAMPLE_FACTORS           	-	P10_3	Tissue:Skin	
SUBJECT_SAMPLE_FACTORS           	-	P14_1	Tissue:Skin	
SUBJECT_SAMPLE_FACTORS           	-	P14_2	Tissue:Skin	
SUBJECT_SAMPLE_FACTORS           	-	P14_3	Tissue:Skin	
SUBJECT_SAMPLE_FACTORS           	-	P15_1	Tissue:Skin	
SUBJECT_SAMPLE_FACTORS           	-	P15_2	Tissue:Skin	
SUBJECT_SAMPLE_FACTORS           	-	P15_3	Tissue:Skin	
#COLLECTION
CO:COLLECTION_SUMMARY            	Environmental samples.
CO:SAMPLE_TYPE                   	Skin and Blubber
CO:COLLECTION_METHOD             	NA
CO:COLLECTION_TUBE_TEMP          	4 C
#TREATMENT
TR:TREATMENT_SUMMARY             	None.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	A portion (about 20 mg) of each tissue extract was dissolved in 500 ml of
SP:SAMPLEPREP_SUMMARY            	phosphate buffer (pH = 7.0) and then 500 ml of deuterium oxide containing 0.05 %
SP:SAMPLEPREP_SUMMARY            	v/v trimethylsilane was added.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	-
CH:INSTRUMENT_NAME               	-
CH:COLUMN_NAME                   	-
#ANALYSIS
AN:DATA_FORMAT                   	.fid
AN:ANALYSIS_TYPE                 	NMR
#NMR
NM:INSTRUMENT_NAME               	Varian 600 MHz AR Premium Compact
NM:INSTRUMENT_TYPE               	FT-NMR
NM:NMR_EXPERIMENT_TYPE           	1D-1H
NM:SPECTROMETER_FREQUENCY        	600 MHz
NM:NMR_PROBE                     	OneProbe
NM:NMR_SOLVENT                   	D2O
NM:NMR_TUBE_SIZE                 	5 mm
NM:SHIMMING_METHOD               	Gradient shimming
NM:PULSE_SEQUENCE                	PRESAT
NM:WATER_SUPPRESSION             	Yes, satmode
NM:PULSE_WIDTH                   	5.90 (90°)
NM:POWER_LEVEL                   	63
NM:RECEIVER_GAIN                 	20
NM:PRESATURATION_POWER_LEVEL     	20
NM:CHEMICAL_SHIFT_REF_CPD        	0 ppm (TSP)
NM:TEMPERATURE                   	25
NM:NUMBER_OF_SCANS               	128
NM:ACQUISITION_TIME              	3
NM:RELAXATION_DELAY              	2
NM:SPECTRAL_WIDTH                	9615.4
NM:NUM_DATA_POINTS_ACQUIRED      	64000
NM:REAL_DATA_POINTS              	57692
NM:CHEMICAL_SHIFT_REF_STD        	TSP
#NMR_METABOLITE_DATA
NMR_METABOLITE_DATA:UNITS        	Relative abundance
NMR_METABOLITE_DATA_START
Samples	G7_1	G7_2	G7_3	G10_2	G10_3	G14_1	G14_2	G14_3	G15_1	G15_2	G15_3	P7_1	P7_2	P7_3	P10_2	P10_3	P14_1	P14_2	P14_3	P15_1	P15_2	P15_3
Factors	Tissue:Blubber	Tissue:Blubber	Tissue:Blubber	Tissue:Blubber	Tissue:Blubber	Tissue:Blubber	Tissue:Blubber	Tissue:Blubber	Tissue:Blubber	Tissue:Blubber	Tissue:Blubber	Tissue:Skin	Tissue:Skin	Tissue:Skin	Tissue:Skin	Tissue:Skin	Tissue:Skin	Tissue:Skin	Tissue:Skin	Tissue:Skin	Tissue:Skin	Tissue:Skin
Succinate	318	246.2	229	12.5	4.6	5.8	0	11	0	3.6	1.7	1.8	1.7	4.5	0	4.2	30.8	5.7	10.4	0	0	0
Acetic acid	3483.3	2619.3	2058.4	123.6	202.9	41.1	11.7	89.9	103.6	37.4	0	0	20.6	1055	233.3	0	38	0.1	17.8	131.2	109.5	109.5
Xanthine	733.8	598	994.6	0	5.1	70.8	32.3	0	0.8	12.4	25.9	0	1.9	383.3	3.3	24.6	143.9	33.9	0	5.9	0	0
Propylene glycol	2.1	0	2.6	1.1	0	0	0	0	0	1.1	0	0	0	0	0.1	0	0.9	0.8	0	0	0	0
2-Hydroxyisovalerate	38.4	87.4	42.7	20.3	1.4	0	0	10.7	30.2	0.1	0	0	0.3	17.6	9.8	0	6.5	2.6	3.8	18.2	5.5	5.5
Fructose	0	0	0	9.4	0	22.7	0	0	55.2	461.4	626.6	0	0	31.8	0	0	0	24.6	84.4	121.6	77.3	77.3
2-Hydroxybutyric acid	69.5	125.6	67.9	305.6	403.6	48.9	0	16	238.6	277.8	261.4	51.5	0	0	138.8	290.8	17	0	48.7	35.9	0	0
Formate	200.2	154.6	128.8	74.6	127.8	1.8	2.8	10.1	52.3	17.9	3.4	20.2	2	34.6	65.5	45.3	7.3	0.9	1.7	33.4	41	41
Hypoxanthine	0.7	0.5	3.8	1.8	1.9	0.2	1.5	0.6	4.8	6.5	5.9	1	0.2	1.4	0.4	0.6	0.7	0.3	0.3	3.2	2.2	2.2
3-Hydroxyisovaleric acid	11.8	0	9.4	3.6	0	1	2.5	3.6	4.9	30.1	23.1	0	0	30.6	0	0	4.2	1.4	0	0.2	0	0
L-Proline	0	0	0	180.4	0	90.6	254.9	120.1	112	0	315.8	0	43.7	0	101	177.2	0	114.3	0	0	0	0
Isobutyric acid	406.1	300.4	231.1	122.4	105.6	110.3	237.5	198.8	360.6	499.8	511.1	128	10.9	203.6	61.4	99.9	109.8	116.1	119.1	264.8	207.9	207.9
L-Leucine	214.5	154	107.7	202.9	188.7	132.8	324.4	230.6	486.4	745.9	657.2	140.5	22.3	114	81.3	172.2	178.2	157.9	173.3	235.1	333.9	333.9
Glycine	317.5	299.2	243.3	206.1	306.4	117.6	448	314.9	439.5	306	269.1	29.3	81.6	318.2	234.6	47.8	164.1	25.7	119.7	408.1	154.3	154.3
Valine	345.5	316.2	198.2	82.5	140.8	100.9	192.1	145.9	413.7	322.6	246.3	128.6	11.8	221.7	46.4	75.8	142.9	133.7	116.2	205.5	189.1	189.1
D-Glucose	46.8	0	451.8	0	6.2	0	0	0	21	0	0	0	0	0	0	0	0	0	223.9	45.4	0	0
Isoleucine	94.6	109.6	50.4	402.8	391.7	172.3	401.9	307.7	888.9	1341.6	1294.9	379.8	28.5	551.5	192.1	284.2	164	158.8	205.2	601.5	422.8	422.8
L-Alanine	236.6	196.2	119.9	199.7	221.8	73.6	174	148.4	174	273	251.4	47.8	21.6	146.3	163.9	222.5	101.4	71.5	65.2	173.3	136.5	136.5
Acetone	58.8	524.6	86.8	33	100.9	6.7	1.9	5.2	9	1.1	5.3	63.8	5.2	228.8	41.8	61.8	33	0.8	32.1	0	0	0
L-Lactic acid	157.8	58.8	62	136.9	133.7	43.4	79.1	93.8	60.6	389.8	217.5	94.6	8.6	297.3	65.3	40.7	71.3	53.4	30	113.3	45.4	45.4
L-Glutamic acid	4138.4	2747.1	2994	1239.2	1561	343.3	1021	544.1	824.9	1279.5	1616.4	2251.8	360.9	2429.1	920.4	1909	462.8	524.8	544.4	787.1	346	346
Glycerol	57.2	359.7	93.8	286.7	306.3	588	1225.9	888.3	1749.4	1103.1	2579.5	510.9	149.7	304	80.1	379.1	595.2	486.1	996.5	762.7	1092	1092
L-Lysine	1009.3	898.6	540.2	69.7	0	95.4	324	120.3	295.1	203	550.5	1316	34.4	0	179.3	249.4	155.6	142.1	187.1	150.9	0	0
Tyrosine	34.8	40.5	36.9	30.6	41.1	14.8	27.1	24.5	29.8	30.3	65.2	24.9	2	53.4	13.7	28	12.8	13.6	6	32.5	36.9	36.9
Creatine	117.9	152.2	62.1	98.3	220.9	23.9	75.3	0	101.1	61.8	154.6	126.6	26.4	0	91.1	97.1	47.7	29.7	110.4	17.3	115.3	115.3
3-Hydroxybutyric acid	42.6	24.2	45.9	55	54.3	83.9	119.3	53.4	39.6	48.8	58.2	57.5	3.3	43.7	19.3	50.1	50.9	31.8	74.8	32.4	23.9	23.9
Creatinine	0	87	0	122.6	24.1	36.9	54.9	86.3	1.9	126.2	33.9	15.7	0	0	0	71.7	118.3	45.3	86.9	17	34.6	34.6
L-Phenylalanine	285.4	200.2	158.5	104.1	138.1	25.8	63.8	47.4	33.8	79	59.5	185.9	8.1	268.6	54.8	110.1	47.6	37.1	33.4	34.8	49.1	49.1
L-Threonine	29.6	34.7	64.1	76.7	55.9	13.3	25.7	28.2	121.9	24.9	153.6	19.2	3.8	197.4	23.2	40.8	16.9	6.2	34.3	78.5	44.4	44.4
Malonate	113	129.2	86.4	0	0	28.4	46.1	17.9	23.1	99.4	352.9	9.3	0	337.6	0	4.6	43.9	7.7	165.9	13.4	82.3	82.3
Dimethylamine	19.7	29	10.2	20.8	19.4	11.6	29.7	24	41.2	36.3	19.6	14	2.1	90.2	8.3	12.3	8.8	12.3	10.8	24.5	11	11
Ethanol	126.3	106.4	101.3	9.3	0	96.2	142.6	47.4	41.1	60.3	0	61.6	5.1	252.6	0	34.3	66.1	23.7	55	49.8	0	0
Isopropyl alcohol	13.6	6.2	18.1	4.5	5.2	5.1	5	9.7	8.2	13.6	13.3	10.2	0.8	30.5	3.2	4.3	6	2.6	1.3	13.2	5.2	5.2
Urea	0	6.1	56.7	5.2	22.7	65.7	142.9	120.9	4.3	103.6	157.3	17.2	7.3	15.8	84.9	24.2	63.3	103.7	0	24.4	225.8	225.8
Methionine	81	0	48	39.7	35.8	11.7	10.7	22.9	13.9	35.3	29.7	99.5	9.5	86.8	36	57	24	10.3	0	5.8	0	0
Myo-inositol	39.1	0	43.4	3.9	0	1.1	24.6	0	93.1	0	338.9	0	0.5	0	0	169	0	0	378.3	0	0	0
L-Histidine	821.1	1427.2	0	3941.8	3208.6	848.7	2108.8	3369.1	3067.8	2737.5	1182.6	3763.6	531.2	5836.5	4177.8	3133.2	1262.8	296.1	484.7	2662.9	2396.8	2396.8
Methanol	10851.9	109799.6	17722.2	17387	17291.1	12581.1	64315.8	17068.3	5461.6	64058.6	71668.3	36794.3	77.4	162013.8	6627.9	34708.2	82658.4	2420	58462	32987	60112	60112
Aspartate	232.2	619.6	106.1	298.4	266.9	42.5	55.1	0	116.5	140.8	207.1	227.3	25.4	1420.4	150.4	151.4	195.9	37.7	134.1	112.5	156.5	156.5
L-Arginine	0	0	0	161.1	476.3	20.5	132.4	27.1	16.3	534.1	441	1083	0	0	179.5	1194.5	0	0	0	0	0	0
Dimethyl sulfone	12.3	57.1	13	30	38.5	12.1	10.6	7.4	15.3	19.9	79.1	39.9	5.4	201.5	27.7	37	19	5.9	41.4	3.1	36.4	36.4
3-Hydroxyisobutyrate	5.6	0	0	152.2	209.6	69.6	56	159.2	134.9	157	218.7	290.3	14.9	601.7	149.1	175.9	77.9	12.2	109.4	158.1	56.2	56.2
Citric acid	105.1	80.3	28.3	50.4	61.6	9.2	37.2	19.7	47.9	52.8	64.6	87	6.2	514.2	34.4	77.2	24.9	8.8	15.9	24.2	17.1	17.1
Betaine	60.2	496.2	125.9	144.3	41.4	94.7	97.7	120.3	25.9	46.7	153	0.4	0	759.3	29.4	138.8	642	0	37	558	367.3	367.3
Choline	0.7	10	8	0	0	0.2	4.1	0.2	0	0	31.4	1	0.7	54.1	0.9	1.1	5.2	0	19.3	4.5	21.1	21.1
L-Ornithine	403	0	0	1219.4	1397.3	182.4	381	362.7	1051.6	1546.7	1427.8	2044.5	221.9	8120	939.6	1837.4	185.2	162.9	257.7	1443.6	811.2	811.2
L-Glutamine	627.5	88.3	236.5	0	183.7	4.6	294.3	0	24	0	0	429.5	0	2195.7	0	165.9	94.2	108.8	0	0.8	125.3	125.3
Pyruvic acid	0	0	0	116.1	0	45.8	73.3	102.4	45.9	114.1	119.7	252.2	20	653.1	95.5	75.5	90	59.6	51.3	80.2	57.4	57.4
Pyroglutamic acid	46.2	0	14.5	94.6	37.9	0	4.3	0	16.2	0	23.9	67.6	0.9	428.6	83.8	78.9	0	2.2	24.2	13.6	2	2
Acetoacetate	8.4	0	3.8	36.7	3	14.4	14.5	5.4	17.4	50.1	13.7	71	3.4	392.5	5.4	30.5	47.7	1.4	38.4	2.3	10	10
Carnitine	6.8	0	35.1	0	0	2.4	7	0.7	0	2.4	53.8	0	0	390.3	0	0	0	7.7	14.4	9.8	18.4	18.4
NMR_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name
Succinate
Acetic acid
Xanthine
Propylene glycol
2-Hydroxyisovalerate
Fructose
2-Hydroxybutyric acid
Formate
Hypoxanthine
3-Hydroxyisovaleric acid
L-Proline
Isobutyric acid
L-Leucine
Glycine
Valine
D-Glucose
Isoleucine
L-Alanine
Acetone
L-Lactic acid
L-Glutamic acid
Glycerol
L-Lysine
Tyrosine
Creatine
3-Hydroxybutyric acid
Creatinine
L-Phenylalanine
L-Threonine
Malonate
Dimethylamine
Ethanol
Isopropyl alcohol
Urea
Methionine
Myo-inositol
L-Histidine
Methanol
Aspartate
L-Arginine
Dimethyl sulfone
3-Hydroxyisobutyrate
Citric acid
Betaine
Choline
L-Ornithine
L-Glutamine
Pyruvic acid
Pyroglutamic acid
Acetoacetate
Carnitine
METABOLITES_END
#END