#METABOLOMICS WORKBENCH sdhungana_20140606_9260131_mwtab.txt DATATRACK_ID:105 3. BECK_Urine-Genetic_HILIC MetaData and Analytical Metadata.xlsm DATATRACK_ID:105 STUDY_ID:ST000071 ANALYSIS_ID:AN000112 VERSION 1 CREATED_ON 03-03-2022 #PROJECT PR:PROJECT_TITLE Metabolomic profiling of influenza: a 2009 pandemic H1N1 influenza in lean and PR:PROJECT_TITLE obese mice PR:PROJECT_TYPE Effects and role of obesity on the severity of influenza PR:PROJECT_SUMMARY During the 2009 H1N1 influenza pandemic outbreak, obese individuals were PR:PROJECT_SUMMARY reported to be at greater risk for morbidity and mortality from pandemic PR:PROJECT_SUMMARY infection. However, the mechanisms contributing to greater influenza severity in PR:PROJECT_SUMMARY infected obese individuals remain unclear. Given that one in ten individuals is PR:PROJECT_SUMMARY obese, and worldwide influenza outbreaks are a consistent public health burden, PR:PROJECT_SUMMARY garnering a better understanding of the pathways and mechanisms contributing to PR:PROJECT_SUMMARY greater influenza severity in the obese is essential for limiting influenza PR:PROJECT_SUMMARY infection mortality in this at-risk population. Closely paralleling pH1N1 PR:PROJECT_SUMMARY infection outcome in humans, obese mice exhibit increased morbidity and PR:PROJECT_SUMMARY mortality following pH1N1 infection. In mice, obesity impairs the function of PR:PROJECT_SUMMARY natural killer cells, dendritic cells, macrophage, B cells and memory T cells. PR:PROJECT_SUMMARY Further, several analyses of lung antiviral responses revealed that obese mice PR:PROJECT_SUMMARY have greater lung damage, lung immune cell infiltration and impaired lung PR:PROJECT_SUMMARY healing after infection. Nevertheless, it remains unclear how altered immune PR:PROJECT_SUMMARY cell function contributes to greater lung damage and increased infection PR:PROJECT_SUMMARY severity in obese mice. Metabolomics will be used to dissect the metabolic PR:PROJECT_SUMMARY consequences of obesity on the immune response to pH1N1 infection. We will PR:PROJECT_SUMMARY compare metabolic profiles of lung-specific and peripheral samples from PR:PROJECT_SUMMARY uninfected and infected lean and obese mice during early and late phases of PR:PROJECT_SUMMARY influenza immunity. PR:INSTITUTE University of North Carolina at Chapel Hill PR:DEPARTMENT Department of Nutrition PR:LAST_NAME Beck PR:FIRST_NAME Melinda PR:ADDRESS 2303 MHRB, CB #7461, UNC, Chapel Hill NC 27599 PR:EMAIL melinda_beck@unc.edu PR:PHONE 919-966-6809 PR:DOI http://dx.doi.org/10.21228/M8QG6H #STUDY ST:STUDY_TITLE Metabolomic profiling of influenza: a 2009 pandemic H1N1 influenza in lean and ST:STUDY_TITLE obese mice (via Urine) ST:INSTITUTE University of North Carolina ST:DEPARTMENT Systems and Translational Sciences ST:LABORATORY Sumner Lab ST:LAST_NAME Sumner ST:FIRST_NAME Susan ST:ADDRESS Eastern Regional Comprehensive Metabolomics Resource Core, UNC Nutrition ST:ADDRESS Research Institute, 500 Laureate Way, Kannapolis, NC, 28081 ST:EMAIL susan_sumner@unc.edu ST:PHONE 704-250-5066 ST:SUBMIT_DATE 2014-06-14 #SUBJECT SU:SUBJECT_TYPE Animal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENOTYPE_STRAIN C57BL/6J SU:AGE_OR_AGE_RANGE approx. 15-16 weeks SU:GENDER male and female SU:ANIMAL_ANIMAL_SUPPLIER The Jackson Laboratory SU:ANIMAL_FEED high fat, low fat or normal chow SU:SPECIES_GROUP Mammal #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - UPALL0001 Phenotype:-- | Infection Status:-- | Sex:-- SUBJECT_SAMPLE_FACTORS - UPALL0002 Phenotype:-- | Infection Status:-- | Sex:-- SUBJECT_SAMPLE_FACTORS - UPALL0003 Phenotype:-- | Infection Status:-- | Sex:-- SUBJECT_SAMPLE_FACTORS - LUI2_67 Phenotype:lean | Infection Status:D2 Infected | Sex:f SUBJECT_SAMPLE_FACTORS - LUI2_68 Phenotype:lean | Infection Status:D2 Infected | Sex:f SUBJECT_SAMPLE_FACTORS - LUI2_92 Phenotype:lean | Infection Status:D2 Infected | Sex:f SUBJECT_SAMPLE_FACTORS - 65 Phenotype:lean | Infection Status:D8 infected | Sex:m SUBJECT_SAMPLE_FACTORS - 86 Phenotype:lean | Infection Status:D8 infected | Sex:m SUBJECT_SAMPLE_FACTORS - 94 Phenotype:lean | Infection Status:D8 infected | Sex:m SUBJECT_SAMPLE_FACTORS - LUC_67 Phenotype:lean | Infection Status:uninfected | Sex:f SUBJECT_SAMPLE_FACTORS - LUC_68 Phenotype:lean | Infection Status:uninfected | Sex:f SUBJECT_SAMPLE_FACTORS - LUC_92 Phenotype:lean | Infection Status:uninfected | Sex:f SUBJECT_SAMPLE_FACTORS - 36 Phenotype:lean | Infection Status:uninfected | Sex:m SUBJECT_SAMPLE_FACTORS - 47 Phenotype:lean | Infection Status:uninfected | Sex:m SUBJECT_SAMPLE_FACTORS - 51 Phenotype:lean | Infection Status:uninfected | Sex:m SUBJECT_SAMPLE_FACTORS - OUI2_74 Phenotype:obese | Infection Status:D2 Infected | Sex:f SUBJECT_SAMPLE_FACTORS - OUI2_75 Phenotype:obese | Infection Status:D2 Infected | Sex:f SUBJECT_SAMPLE_FACTORS - OUI2_91 Phenotype:obese | Infection Status:D2 Infected | Sex:f SUBJECT_SAMPLE_FACTORS - 109 Phenotype:obese | Infection Status:D8 infected | Sex:m SUBJECT_SAMPLE_FACTORS - 88 Phenotype:obese | Infection Status:D8 infected | Sex:m SUBJECT_SAMPLE_FACTORS - OUC_74 Phenotype:obese | Infection Status:uninfected | Sex:f SUBJECT_SAMPLE_FACTORS - OUC_75 Phenotype:obese | Infection Status:uninfected | Sex:f SUBJECT_SAMPLE_FACTORS - OUC_91 Phenotype:obese | Infection Status:uninfected | Sex:f SUBJECT_SAMPLE_FACTORS - 35 Phenotype:obese | Infection Status:uninfected | Sex:m SUBJECT_SAMPLE_FACTORS - 53 Phenotype:obese | Infection Status:uninfected | Sex:m #COLLECTION CO:SAMPLE_TYPE Tissue CO:STORAGE_CONDITIONS -80 #TREATMENT #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Samples were homogenized on a baed beater at1750rpm in 10uL of 50:50 SP:SAMPLEPREP_SUMMARY acetonitrile:water per 1 mg of tissue using washed ceramic beads. Extraction was SP:SAMPLEPREP_SUMMARY performed using acetonitrile. Samples were dried down and reconstituted in SP:SAMPLEPREP_SUMMARY 100uL of 95:5 water:methanol. SP:SAMPLEPREP_PROTOCOL_FILENAME RTI_BECK_Urine-Genetic_HILIC_Metabolomics_Procedure.docx SP:PROCESSING_METHOD Homogenization SP:EXTRACTION_METHOD Acetonitrile SP:EXTRACT_STORAGE -80 C SP:SAMPLE_RESUSPENSION 95:5 Water:Methanol SP:SAMPLE_SPIKING L-Tryptophan-d5 SP:ORGAN Lung #CHROMATOGRAPHY CH:INSTRUMENT_NAME Waters Acquity CH:COLUMN_NAME Waters Acquity BEH Amide (150 x 2.1mm, 1.7um) CH:COLUMN_PRESSURE 6000-10000 CH:COLUMN_TEMPERATURE 40 C CH:FLOW_GRADIENT Time(min) Flow Rate %A %B: 1. Initial 0.400 99.0 1.0; 2. 1.00 0.400 99.0 1.0; 3. CH:FLOW_GRADIENT 12.00 0.400 1.0 99.0; 4. 14.00 0.400 1.0 99.0; 5. 15.00 0.400 99.0 1.0; 6. 19.00 CH:FLOW_GRADIENT 0.400 99.0 1.0 CH:FLOW_RATE 0.4 mL/min CH:INJECTION_TEMPERATURE 8 C CH:INTERNAL_STANDARD L-Tryptophan-d5 CH:SOLVENT_A 10mM ammonium acetate in 95/5 acetonitrile/water with 0.1% formic acid CH:SOLVENT_B 10mM ammonium acetate in 50/50 acetonitrile/water with 0.1% formic acid CH:ANALYTICAL_TIME 19 min CH:WEAK_WASH_SOLVENT_NAME 5%MeOH CH:WEAK_WASH_VOLUME 1000 uL CH:STRONG_WASH_SOLVENT_NAME 80%MeOH CH:STRONG_WASH_VOLUME 1000 uL CH:TARGET_SAMPLE_TEMPERATURE 8 C CH:SAMPLE_LOOP_SIZE 10 uL CH:SAMPLE_SYRINGE_SIZE 100 uL CH:RANDOMIZATION_ORDER Yes CH:CHROMATOGRAPHY_TYPE HILIC #ANALYSIS AN:LABORATORY_NAME RTI AN:ANALYSIS_TYPE MS AN:ACQUISITION_DATE 41656 AN:SOFTWARE_VERSION MassLynx 4.1 AN:OPERATOR_NAME Suraj Dhungana, Darya Cheng AN:DETECTOR_TYPE TOF (negative) #MS MS:INSTRUMENT_NAME Waters Synapt G2 QTOF MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:CAPILLARY_TEMPERATURE 110 C MS:CAPILLARY_VOLTAGE 2.8 kV MS:COLLISION_ENERGY 4 MS:FRAGMENTATION_METHOD CID MS:HELIUM_FLOW 180 MS:IONIZATION ES- MS:SOURCE_TEMPERATURE 110 C MS:DATAFORMAT Continuum MS:DESOLVATION_GAS_FLOW 400 L/Hr MS:DESOLVATION_TEMPERATURE 400 C MS:RESOLUTION_SETTING 18000 MS:SCAN_RANGE_MOVERZ 50-1000 m/z MS:SCANNING_CYCLE 1 s MS:TUBE_LENS_VOLTAGE 74 MS:MS_RESULTS_FILE ST000071_AN000112_Results.txt UNITS:Peak area Has m/z:Yes Has RT:Yes RT units:Minutes #END