#METABOLOMICS WORKBENCH takis_20180915_113113 DATATRACK_ID:1509 STUDY_ID:ST001054 ANALYSIS_ID:AN001723 PROJECT_ID:PR000707 VERSION 1 CREATED_ON September 17, 2018, 5:25 pm #PROJECT PR:PROJECT_TITLE Fingerprinting gastrointestinal diseases by 1H NMR PR:PROJECT_SUMMARY A blood sample was drawn from each patient admitted to the Florence main PR:PROJECT_SUMMARY hospital emergency room with severe abdominal pain. The corresponding sera PR:PROJECT_SUMMARY underwent 1H NMR metabolomics fingerprinting to discriminate patients with PR:PROJECT_SUMMARY symptoms of upper vs. diffuse abdominal/intestinal pain by the whole 1H NMR PR:PROJECT_SUMMARY serum fingerprint and use it as a first-line discriminator for these symptoms PR:PROJECT_SUMMARY and the diseases behind them. PR:INSTITUTE Giotto Biotech s.r.l. PR:LAST_NAME Takis PR:FIRST_NAME Panteleimon PR:ADDRESS Via Madonna del Piano 6, 50019 Sesto Fiorentino (FI), Italy PR:EMAIL takis@giottobiotech.com PR:PHONE +393423233750 #STUDY ST:STUDY_TITLE Fingerprinting gastrointestinal diseases by 1H NMR ST:STUDY_SUMMARY We studied 64 patients admitted to the Florence main hospital emergency room ST:STUDY_SUMMARY with severe abdominal pain. A blood sample was drawn from each patient at ST:STUDY_SUMMARY admission, and the corresponding sera underwent 1H NMR metabolomics ST:STUDY_SUMMARY fingerprinting. Unsupervised PCA analysis showed a significant discrimination ST:STUDY_SUMMARY between a group of patients with symptoms of upper abdominal pain and a second ST:STUDY_SUMMARY group consisting of patients with diffuse abdominal/intestinal pain. Prompted by ST:STUDY_SUMMARY this observation, supervised statistical analysis (OPLS-DA) showed a very good ST:STUDY_SUMMARY discrimination (> 90 %) between the two groups of symptoms. Actually in the ST:STUDY_SUMMARY present study, upper abdominal pain may result from either symptomatic ST:STUDY_SUMMARY gallstones, cholecystitis or pancreatitis, while diffuse abdominal/intestinal ST:STUDY_SUMMARY pain may result from either intestinal ischemia, strangulated obstruction or ST:STUDY_SUMMARY mechanical obstruction. Although limited by the small number of samples from ST:STUDY_SUMMARY each of these six conditions, discrimination of these diseases was attempted. In ST:STUDY_SUMMARY the first symptom group, > 70% discrimination accuracy was obtained among ST:STUDY_SUMMARY symptomatic gallstones, pancreatitis and cholecystitis, while for the second ST:STUDY_SUMMARY symptom group > 85% classification accuracy was obtained for intestinal ST:STUDY_SUMMARY ischemia, strangulated obstruction and mechanical obstruction. No single ST:STUDY_SUMMARY metabolite stands up as a possible biomarker for any of these diseases, while ST:STUDY_SUMMARY the contribution of the whole 1H NMR serum fingerprint seems to be a promising ST:STUDY_SUMMARY candidate, to be confirmed on larger cohorts, as a first-line discriminator for ST:STUDY_SUMMARY these diseases. ST:INSTITUTE Giotto Biotech s.r.l. ST:LAST_NAME Takis ST:FIRST_NAME Panteleimon ST:ADDRESS Via Madonna del Piano 6 ST:EMAIL takis@giottobiotech.com ST:PHONE +393423233750 #SUBJECT SU:SUBJECT_TYPE Human SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 SU:AGE_OR_AGE_RANGE 50-90 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 MAP003 MAP003 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Ischemia SUBJECT_SAMPLE_FACTORS MAP011 MAP011 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Symptomatic Gallstones SUBJECT_SAMPLE_FACTORS MAP013 MAP013 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Strangulated Obstruction SUBJECT_SAMPLE_FACTORS MAP030 MAP030 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Cholecystitis SUBJECT_SAMPLE_FACTORS MAP031 MAP031 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Pancreatitis SUBJECT_SAMPLE_FACTORS MAP036 MAP036 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Ischemia SUBJECT_SAMPLE_FACTORS MAP046 MAP046 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Strangulated Obstruction SUBJECT_SAMPLE_FACTORS MAP050 MAP050 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Symptomatic Gallstones SUBJECT_SAMPLE_FACTORS MAP058 MAP058 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Pancreatitis SUBJECT_SAMPLE_FACTORS MAP066 MAP066 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Pancreatitis SUBJECT_SAMPLE_FACTORS MAP072 MAP072 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Strangulated Obstruction SUBJECT_SAMPLE_FACTORS MAP088 MAP088 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Ischemia SUBJECT_SAMPLE_FACTORS MAP116 MAP116 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Mechanical Obstruction SUBJECT_SAMPLE_FACTORS MAP111 MAP111 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Symptomatic Gallstones SUBJECT_SAMPLE_FACTORS MAP171 MAP171 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Symptomatic Gallstones SUBJECT_SAMPLE_FACTORS MAP177 MAP177 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Mechanical Obstruction SUBJECT_SAMPLE_FACTORS MAP208 MAP208 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Strangulated Obstruction SUBJECT_SAMPLE_FACTORS MAP227 MAP227 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Strangulated Obstruction SUBJECT_SAMPLE_FACTORS MAP246 MAP246 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Strangulated Obstruction SUBJECT_SAMPLE_FACTORS MAP277 MAP277 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Mechanical Obstruction SUBJECT_SAMPLE_FACTORS MAP317 MAP317 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Mechanical Obstruction SUBJECT_SAMPLE_FACTORS MAP338 MAP338 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Mechanical Obstruction SUBJECT_SAMPLE_FACTORS MAP268 MAP268 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Mechanical Obstruction SUBJECT_SAMPLE_FACTORS MAP281 MAP281 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Mechanical Obstruction SUBJECT_SAMPLE_FACTORS MAP291 MAP291 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Ischemia SUBJECT_SAMPLE_FACTORS MAP035 MAP035 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Mechanical Obstruction SUBJECT_SAMPLE_FACTORS MAP044 MAP044 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Cholecystitis SUBJECT_SAMPLE_FACTORS MAP183 MAP183 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Mechanical Obstruction SUBJECT_SAMPLE_FACTORS MAP278 MAP278 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Cholecystitis SUBJECT_SAMPLE_FACTORS MAP217 MAP217 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Mechanical Obstruction SUBJECT_SAMPLE_FACTORS MAP331 MAP331 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Pancreatitis SUBJECT_SAMPLE_FACTORS MAP264 MAP264 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Mechanical Obstruction SUBJECT_SAMPLE_FACTORS MAP271 MAP271 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Ischemia SUBJECT_SAMPLE_FACTORS MAP280 MAP280 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Ischemia SUBJECT_SAMPLE_FACTORS MAP306 MAP306 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Ischemia SUBJECT_SAMPLE_FACTORS MAP005 MAP005 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Symptomatic Gallstones SUBJECT_SAMPLE_FACTORS MAP121 MAP121 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Ischemia SUBJECT_SAMPLE_FACTORS MAP130 MAP130 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Mechanical Obstruction SUBJECT_SAMPLE_FACTORS MAP152 MAP152 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Cholecystitis SUBJECT_SAMPLE_FACTORS MAP154 MAP154 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Cholecystitis SUBJECT_SAMPLE_FACTORS MAP209 MAP209 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Cholecystitis SUBJECT_SAMPLE_FACTORS MAP212 MAP212 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Mechanical Obstruction SUBJECT_SAMPLE_FACTORS MAP222 MAP222 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Symptomatic Gallstones SUBJECT_SAMPLE_FACTORS MAP228 MAP228 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Symptomatic Gallstones SUBJECT_SAMPLE_FACTORS MAP230 MAP230 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Pancreatitis SUBJECT_SAMPLE_FACTORS MAP232 MAP232 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Symptomatic Gallstones SUBJECT_SAMPLE_FACTORS MAP239 MAP239 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Symptomatic Gallstones SUBJECT_SAMPLE_FACTORS MAP244 MAP244 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Mechanical Obstruction SUBJECT_SAMPLE_FACTORS MAP250 MAP250 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Strangulated Obstruction SUBJECT_SAMPLE_FACTORS MAP258 MAP258 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Mechanical Obstruction SUBJECT_SAMPLE_FACTORS MAP295 MAP295 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Pancreatitis SUBJECT_SAMPLE_FACTORS MAP308 MAP308 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Cholecystitis SUBJECT_SAMPLE_FACTORS MAP311 MAP311 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Pancreatitis SUBJECT_SAMPLE_FACTORS MAP315 MAP315 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Cholecystitis SUBJECT_SAMPLE_FACTORS MAP329 MAP329 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Symptomatic Gallstones SUBJECT_SAMPLE_FACTORS MAP024 MAP024 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Strangulated Obstruction SUBJECT_SAMPLE_FACTORS MAP049 MAP049 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Mechanical Obstruction SUBJECT_SAMPLE_FACTORS MAP101 MAP101 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Pancreatitis SUBJECT_SAMPLE_FACTORS MAP117 MAP117 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Ischemia SUBJECT_SAMPLE_FACTORS MAP118 MAP118 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Mechanical Obstruction SUBJECT_SAMPLE_FACTORS MAP119 MAP119 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Symptomatic Gallstones SUBJECT_SAMPLE_FACTORS MAP158 MAP158 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Symptomatic Gallstones SUBJECT_SAMPLE_FACTORS MAP210 MAP210 Abdominal pain location (symptom):Diffuse abdominal/intestinal pain | clinical diagnosis:Intestinal Mechanical Obstruction SUBJECT_SAMPLE_FACTORS MAP296 MAP296 Abdominal pain location (symptom):Upper abdominal pain | clinical diagnosis:Cholecystitis #COLLECTION CO:COLLECTION_SUMMARY Blood was drawn at admission to the emergency room, after acquiring the CO:COLLECTION_SUMMARY appropriate informed consent (explaining to the patients the aims of the study CO:COLLECTION_SUMMARY and ensuring them the complete anonymity of the data collected). Each sample CO:COLLECTION_SUMMARY consisted of 8-10 cc of peripheral blood and was immediately ultra-centrifuged CO:COLLECTION_SUMMARY for about 10-15 minutes at 1300 RCF. Serum samples were then subdivided in small CO:COLLECTION_SUMMARY aliquots (about 500-600 microliters each), and immediately stored at -80 °C. CO:COLLECTION_PROTOCOL_FILENAME Blood_collection.docx CO:SAMPLE_TYPE Blood (serum) CO:STORAGE_CONDITIONS Described in summary #TREATMENT TR:TREATMENT_SUMMARY No treatment #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Frozen serum samples were thawed at room temperature and 300 μl of each sample SP:SAMPLEPREP_SUMMARY were added to a total of 300 μl of a sodium phosphate buffer at pH 7.4 (75 mM SP:SAMPLEPREP_SUMMARY Na2HPO4, 20 % (v/v) D2O, 0.025 % (v/v) NaN3 and 0.8 % (w/v) sodium SP:SAMPLEPREP_SUMMARY trimethylsilyl[2,2,3,3-22H4]propionate (TMSP)). From the mixture, 450 μl were SP:SAMPLEPREP_SUMMARY transferred into a 4.25 mm NMR disposable tube, acquired from Bruker BioSpin SP:SAMPLEPREP_SUMMARY srl. SP:SAMPLEPREP_PROTOCOL_FILENAME NMR_Sample_preparation.docx SP:PROCESSING_STORAGE_CONDITIONS Room temperature #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE - CH:INSTRUMENT_NAME - CH:COLUMN_NAME - #ANALYSIS AN:ANALYSIS_TYPE NMR AN:ANALYSIS_PROTOCOL_FILE NMR_details.docx AN:DATA_FORMAT 1r #NMR NM:INSTRUMENT_NAME Bruker Avance III NM:INSTRUMENT_TYPE FT-NMR NM:NMR_EXPERIMENT_TYPE 1D-1H NM:SPECTROMETER_FREQUENCY 600 MHz NM:NMR_SOLVENT Serum D2O Buffer NM:NMR_TUBE_SIZE 4.25 mm NM:TEMPERATURE 36.85 NM:NUMBER_OF_SCANS 64 NM:DUMMY_SCANS 4 NM:ACQUISITION_TIME 2.7 NM:RELAXATION_DELAY 4 NM:LINE_BROADENING 1 Hz NM:CHEMICAL_SHIFT_REF_STD TMSP at 0 ppm NM:BINNED_INCREMENT 0.02 NM:BINNED_DATA_EXCLUDED_RANGE 4.40-5.00 (ppm) H2O region NM:NMR_RESULTS_FILE ST001054_AN001723_Results.txt UNITS:NMR bins #END