#METABOLOMICS WORKBENCH dhananjay_shinde5_20240914_131224 DATATRACK_ID:5191 STUDY_ID:ST003482 ANALYSIS_ID:AN005718 PROJECT_ID:PR002139 VERSION 1 CREATED_ON September 14, 2024, 3:09 pm #PROJECT PR:PROJECT_TITLE Tissue niche influences immune and metabolic profiles to Staphylococcus aureus PR:PROJECT_TITLE biofilm infection PR:PROJECT_TYPE Intracellular data PR:PROJECT_SUMMARY Infection is a devastating post-surgical complication, often requiring PR:PROJECT_SUMMARY additional procedures and prolonged antibiotic therapy. This is especially PR:PROJECT_SUMMARY relevant for craniotomy and prosthetic joint infections (PJI), both of which are PR:PROJECT_SUMMARY characterized by biofilm formation on the bone or implant surface, respectively, PR:PROJECT_SUMMARY with S. aureus representing a primary cause. The local tissue microenvironment PR:PROJECT_SUMMARY likely has profound effects on immune attributes that can influence treatment PR:PROJECT_SUMMARY efficacy, which becomes critical to consider when developing therapeutics for PR:PROJECT_SUMMARY biofilm infections. However, the extent to which distinct tissue niches PR:PROJECT_SUMMARY influence immune function during biofilm development remains relatively unknown. PR:PROJECT_SUMMARY To address this, we compare the metabolomic, transcriptomic, and functional PR:PROJECT_SUMMARY attributes of leukocytes in mouse models of S. aureus craniotomy and PJI PR:PROJECT_SUMMARY complemented with patient samples from both infection modalities, which reveals PR:PROJECT_SUMMARY profound tissue niche-dependent differences in nucleic acid, amino acid, and PR:PROJECT_SUMMARY lipid metabolism with links to immune modulation. These signatures are both PR:PROJECT_SUMMARY spatially and temporally distinct, differing not only between infection sites PR:PROJECT_SUMMARY but evolving over time within a single model. Collectively, this demonstrates PR:PROJECT_SUMMARY that biofilms elicit unique immune and metabolic responses that are heavily PR:PROJECT_SUMMARY influenced by the local tissue microenvironment, which will likely have PR:PROJECT_SUMMARY important implications when designing therapeutic approaches targeting these PR:PROJECT_SUMMARY infections. PR:INSTITUTE UNMC PR:DEPARTMENT Department of pathology, microbiology and immunology PR:LABORATORY Pathology, Microbiology and Immunology PR:LAST_NAME Dhananjay PR:FIRST_NAME Shinde PR:ADDRESS DRC 2, 7066, UNMC, Emily st PR:EMAIL dhananjay.shinde@unmc.edu PR:PHONE 4025597623 #STUDY ST:STUDY_TITLE Tissue niche influences immune and metabolic profiles to Staphylococcus aureus ST:STUDY_TITLE biofilm infection ST:STUDY_SUMMARY Infection is a devastating post-surgical complication, often requiring ST:STUDY_SUMMARY additional procedures and prolonged antibiotic therapy. This is especially ST:STUDY_SUMMARY relevant for craniotomy and prosthetic joint infections (PJI), both of which are ST:STUDY_SUMMARY characterized by biofilm formation on the bone or implant surface, respectively, ST:STUDY_SUMMARY with S. aureus representing a primary cause. The local tissue microenvironment ST:STUDY_SUMMARY likely has profound effects on immune attributes that can influence treatment ST:STUDY_SUMMARY efficacy, which becomes critical to consider when developing therapeutics for ST:STUDY_SUMMARY biofilm infections. However, the extent to which distinct tissue niches ST:STUDY_SUMMARY influence immune function during biofilm development remains relatively unknown. ST:STUDY_SUMMARY To address this, we compare the metabolomic, transcriptomic, and functional ST:STUDY_SUMMARY attributes of leukocytes in mouse models of S. aureus craniotomy and PJI ST:STUDY_SUMMARY complemented with patient samples from both infection modalities, which reveals ST:STUDY_SUMMARY profound tissue niche-dependent differences in nucleic acid, amino acid, and ST:STUDY_SUMMARY lipid metabolism with links to immune modulation. These signatures are both ST:STUDY_SUMMARY spatially and temporally distinct, differing not only between infection sites ST:STUDY_SUMMARY but evolving over time within a single model. Collectively, this demonstrates ST:STUDY_SUMMARY that biofilms elicit unique immune and metabolic responses that are heavily ST:STUDY_SUMMARY influenced by the local tissue microenvironment, which will likely have ST:STUDY_SUMMARY important implications when designing therapeutic approaches targeting these ST:STUDY_SUMMARY infections. ST:INSTITUTE UNMC ST:DEPARTMENT Department of pathology, microbiology and immunology ST:LAST_NAME Shinde ST:FIRST_NAME Dhananjay ST:ADDRESS DRC 2, 7066, UNMC, Emily st ST:EMAIL dhananjay.shinde@unmc.edu ST:PHONE 4025597623 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:GENDER Male and female #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 - d3-Crain_1 Sample source:Galea | Experimental Variable:d3 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-Crain_1.mzXML SUBJECT_SAMPLE_FACTORS - d3-Crain_2 Sample source:Galea | Experimental Variable:d3 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-Crain_2.mzXML SUBJECT_SAMPLE_FACTORS - d3-Crain_3 Sample source:Galea | Experimental Variable:d3 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-Crain_3.mzXML SUBJECT_SAMPLE_FACTORS - d3-Crain_4 Sample source:Galea | Experimental Variable:d3 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-Crain_4.mzXML SUBJECT_SAMPLE_FACTORS - d3-Crain_5 Sample source:Galea | Experimental Variable:d3 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-Crain_5.mzXML SUBJECT_SAMPLE_FACTORS - d3-PJI_1 Sample source:Tissue (PJI) | Experimental Variable:d3 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-PJI_1.mzXML SUBJECT_SAMPLE_FACTORS - d3-PJI_2 Sample source:Tissue (PJI) | Experimental Variable:d3 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-PJI_2.mzXML SUBJECT_SAMPLE_FACTORS - d3-PJI_3 Sample source:Tissue (PJI) | Experimental Variable:d3 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-PJI_3.mzXML SUBJECT_SAMPLE_FACTORS - d3-PJI_4 Sample source:Tissue (PJI) | Experimental Variable:d3 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-PJI_4.mzXML SUBJECT_SAMPLE_FACTORS - d3-PJI_5 Sample source:Tissue (PJI) | Experimental Variable:d3 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-PJI_5.mzXML SUBJECT_SAMPLE_FACTORS - d7-Crain_1 Sample source:Galea | Experimental Variable:d7 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-Crain_1.mzXML SUBJECT_SAMPLE_FACTORS - d7-Crain_2 Sample source:Galea | Experimental Variable:d7 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-Crain_2.mzXML SUBJECT_SAMPLE_FACTORS - d7-Crain_3 Sample source:Galea | Experimental Variable:d7 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-Crain_3.mzXML SUBJECT_SAMPLE_FACTORS - d7-Crain_4 Sample source:Galea | Experimental Variable:d7 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-Crain_4.mzXML SUBJECT_SAMPLE_FACTORS - d7-Crain_5 Sample source:Galea | Experimental Variable:d7 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-Crain_5.mzXML SUBJECT_SAMPLE_FACTORS - d7-PJI_1 Sample source:Tissue (PJI) | Experimental Variable:d7 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-PJI_1.mzXML SUBJECT_SAMPLE_FACTORS - d7-PJI_2 Sample source:Tissue (PJI) | Experimental Variable:d7 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-PJI_2.mzXML SUBJECT_SAMPLE_FACTORS - d7-PJI_3 Sample source:Tissue (PJI) | Experimental Variable:d7 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-PJI_3.mzXML SUBJECT_SAMPLE_FACTORS - d7-PJI_4 Sample source:Tissue (PJI) | Experimental Variable:d7 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-PJI_4.mzXML SUBJECT_SAMPLE_FACTORS - d7-PJI_5 Sample source:Tissue (PJI) | Experimental Variable:d7 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-PJI_5.mzXML SUBJECT_SAMPLE_FACTORS - d14-Crain_1 Sample source:Galea | Experimental Variable:d14 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-Crain_1.mzXML SUBJECT_SAMPLE_FACTORS - d14-Crain_2 Sample source:Galea | Experimental Variable:d14 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-Crain_2.mzXML SUBJECT_SAMPLE_FACTORS - d14-Crain_3 Sample source:Galea | Experimental Variable:d14 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-Crain_3.mzXML SUBJECT_SAMPLE_FACTORS - d14-Crain_4 Sample source:Galea | Experimental Variable:d14 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-Crain_4.mzXML SUBJECT_SAMPLE_FACTORS - d14-Crain_5 Sample source:Galea | Experimental Variable:d14 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-Crain_5.mzXML SUBJECT_SAMPLE_FACTORS - d14-PJI_1 Sample source:Tissue (PJI) | Experimental Variable:d14 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-PJI_1.mzXML SUBJECT_SAMPLE_FACTORS - d14-PJI_2 Sample source:Tissue (PJI) | Experimental Variable:d14 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-PJI_2.mzXML SUBJECT_SAMPLE_FACTORS - d14-PJI_3 Sample source:Tissue (PJI) | Experimental Variable:d14 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-PJI_3.mzXML SUBJECT_SAMPLE_FACTORS - d14-PJI_4 Sample source:Tissue (PJI) | Experimental Variable:d14 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-PJI_4.mzXML SUBJECT_SAMPLE_FACTORS - d14-PJI_5 Sample source:Tissue (PJI) | Experimental Variable:d14 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-PJI_5.mzXML #COLLECTION CO:COLLECTION_SUMMARY The intracellular metabolome of granulocytes isolated from the galea and PJI CO:COLLECTION_SUMMARY tissue of mouse craniotomy and PJI models, respectively, were profiled by CO:COLLECTION_SUMMARY LC-HRMS. Fifteen mice were infected for each biofilm model (craniotomy and PJI) CO:COLLECTION_SUMMARY with single-cell suspensions prepared by pooling tissue homogenates from 3 mice CO:COLLECTION_SUMMARY into one sample, resulting in 5 biological replicates. Samples were then CO:COLLECTION_SUMMARY enriched for granulocytes using Ly6G magnetic beads (Miltenyi Biotec, Bergishch CO:COLLECTION_SUMMARY Gladbach, Germany). Magnetic beads were utilized for purification instead of CO:COLLECTION_SUMMARY FACS to limit alterations in cellular metabolism that may occur with extended CO:COLLECTION_SUMMARY processing times. Samples were washed twice in 1X PBS, resuspended in 80% MetOH, CO:COLLECTION_SUMMARY and frozen at -80°C for 15 min. Metabolomics was also performed on supernatants CO:COLLECTION_SUMMARY prepared from brain, galea, and PJI tissue homogenates at days 3, 7, and 14 CO:COLLECTION_SUMMARY post-infection. Similar to granulocyte intracellular metabolite collection, CO:COLLECTION_SUMMARY tissue supernatants were centrifuged at high speed to remove residual cellular CO:COLLECTION_SUMMARY material, combined with 100% MetOH to yield an 80% MetOH solution, and frozen at CO:COLLECTION_SUMMARY -80°C. CO:SAMPLE_TYPE Extracellular infection sites homogenate, intracellular granulocyte #TREATMENT TR:TREATMENT_SUMMARY All mice were group-housed at 21-23°C (22°C average) and 30-70% humidity (55% TR:TREATMENT_SUMMARY average) under 12 h light/dark cycle with free access to food (2019S Teklad TR:TREATMENT_SUMMARY Global 19% Protein Extruded Rodent Diet; Inotiv, West Lafayette, IN) and water. TR:TREATMENT_SUMMARY S. aureus craniotomy infection was established in male and female 8- to TR:TREATMENT_SUMMARY 10-week-old C57BL/6J mice (RRID:IMSR_JAX:000664). Briefly, ketamine and xylazine TR:TREATMENT_SUMMARY were administered to achieve a surgical plane of anesthesia before skin TR:TREATMENT_SUMMARY disinfection with betadine. A midline incision of the scalp was made, whereupon TR:TREATMENT_SUMMARY a bone flap was created using a high-speed pneumatic drill (Stryker Corporation, TR:TREATMENT_SUMMARY Kalamazoo, MI). The bone flap was incubated with 0.5 mL of S. aureus USA300 TR:TREATMENT_SUMMARY LAC13C diluted to 10^6/mL in brain-heart infusion broth at 37°C for 5 min and TR:TREATMENT_SUMMARY subsequently rinsed in PBS, blotted dry on a sterile field, and reimplanted. TR:TREATMENT_SUMMARY This procedure reliably produces an infectious inoculum of 10^3 CFUs per bone TR:TREATMENT_SUMMARY flap, which more accurately models surgical site infection that progresses to TR:TREATMENT_SUMMARY biofilm formation. The scalp incision was closed using 6-0 nylon suture and TR:TREATMENT_SUMMARY animals were continuously monitored until they regained ambulation, and daily TR:TREATMENT_SUMMARY thereafter. S. aureus PJI was established in male and female 8- to 10-week-old TR:TREATMENT_SUMMARY C57BL/6J mice. Mice were anesthetized with ketamine and xylazine prior to skin TR:TREATMENT_SUMMARY disinfection with betadine and initial incision of the quadriceps. A secondary TR:TREATMENT_SUMMARY incision was made to laterally displace the patellar tendon and a burr hole was TR:TREATMENT_SUMMARY created in the distal end of the femur with a 26-gauge needle. An TR:TREATMENT_SUMMARY orthopedic-grade nickel-titanium Kirschner wire (0.6 mm diameter; Custom Wire TR:TREATMENT_SUMMARY Technologies, Port Washington, WI) was inserted through the burr hole into the TR:TREATMENT_SUMMARY medullary canal and 10^3 CFUs of S. aureus USA300 LAC13C were inoculated at the TR:TREATMENT_SUMMARY tip of the wire and the incision was closed with nylon suture. Mice received TR:TREATMENT_SUMMARY buprenorphine SR to alleviate any post-surgical pain and were continuously TR:TREATMENT_SUMMARY monitored until ambulatory and daily thereafter. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY A 13C15N-labeled canonical amino acid (13C15N-CAA) mix (Cambridge Isotope SP:SAMPLEPREP_SUMMARY Laboratories, Andover, MA) was added to the extraction buffer comprised of 80% SP:SAMPLEPREP_SUMMARY MeOH as an internal standard. Samples were then pelleted, and supernatants were SP:SAMPLEPREP_SUMMARY dried by vacuum centrifuge for storage at -80°C. Cell pellets were stored for SP:SAMPLEPREP_SUMMARY protein quantification to normalize metabolite peak areas. Prior to LC-HRMS, SP:SAMPLEPREP_SUMMARY samples were reconstituted in 100 µL of 50% MetOH, pelleted, and the resulting SP:SAMPLEPREP_SUMMARY supernatant was analyzed. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Metabolite separation was performed by liquid chromatography using a XBridge CH:CHROMATOGRAPHY_SUMMARY Amide analytical column (150 × 2.1mm ID; 1.7µm particle size; Waters CH:CHROMATOGRAPHY_SUMMARY Corporation, Milford, MA) and a binary solvent system infused at a flow rate of CH:CHROMATOGRAPHY_SUMMARY 0.3 ml/min. A guard XBridge Amide column (20 × 2.1mm ID; 3.5µm particle CH:CHROMATOGRAPHY_SUMMARY size; Waters Corporation) was connected in front of the analytical column. CH:CHROMATOGRAPHY_SUMMARY Mobile phase A was composed of ammonium acetate and ammonium hydroxide (10 mM CH:CHROMATOGRAPHY_SUMMARY each) containing 5% acetonitrile in LC-MS grade water; mobile phase B was 100% CH:CHROMATOGRAPHY_SUMMARY LC-MS grade acetonitrile. The pH of mobile phase A was adjusted to 8.0 using CH:CHROMATOGRAPHY_SUMMARY glacial acetic acid. The UHPLC pumps were operated in gradient mode. The amide CH:CHROMATOGRAPHY_SUMMARY column was maintained at 40°C, and the autosampler temperature was held at 5°C CH:CHROMATOGRAPHY_SUMMARY throughout data acquisition. The injection volume for all samples was 5 µl with CH:CHROMATOGRAPHY_SUMMARY the 13C15N-CAA mix as the internal standard. CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME Waters ACQUITY UPLC BEH Amide (150 x 2.1mm,1.7um) CH:SOLVENT_A ammonium acetate and ammonium hydroxide (10 mM each) containing 5% acetonitrile CH:SOLVENT_A in LC-MS grade water CH:SOLVENT_B acetonitrile CH:FLOW_GRADIENT B% at initial 85% CH:FLOW_RATE 0.3ml/min CH:COLUMN_TEMPERATURE 40 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Orbitrap Exploris 480 MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS A HRMS Orbitrap (Exploris 480; Thermo Fisher Scientific) was operated in MS:MS_COMMENTS polarity switching mode and was used for untargeted metabolomics in a MS:MS_COMMENTS data-dependent MS/MS acquisition mode (DDA). Electrospray ionization (ESI) MS:MS_COMMENTS parameters were optimized as follows: electrospray ion voltage of -2700V and MS:MS_COMMENTS 3500V in negative and positive mode respectively, ion transfer tube temperature MS:MS_COMMENTS was maintained at 400°C, and m/z scan range was 70-1050 Da. Orbitrap resolution MS:MS_COMMENTS for precursor ion scans were maintained at 240,000 for intracellular and 120,000 MS:MS_COMMENTS for extracellular whereas fragment ion scans were maintained at 120,000 and MS:MS_COMMENTS 60,000, for intracellular and extracellular respectively. Normalized collision MS:MS_COMMENTS energies at 30, 50, and 150% were used for fragmentation and data was acquired MS:MS_COMMENTS in profile mode. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Relative intensities MS_METABOLITE_DATA_START Samples d3-Crain_1 d3-Crain_2 d3-Crain_3 d3-Crain_4 d3-Crain_5 d3-PJI_1 d3-PJI_2 d3-PJI_3 d3-PJI_4 d3-PJI_5 d7-Crain_1 d7-Crain_2 d7-Crain_3 d7-Crain_4 d7-Crain_5 d7-PJI_1 d7-PJI_2 d7-PJI_3 d7-PJI_4 d7-PJI_5 d14-Crain_1 d14-Crain_2 d14-Crain_3 d14-Crain_4 d14-Crain_5 d14-PJI_1 d14-PJI_2 d14-PJI_3 d14-PJI_4 d14-PJI_5 Factors Sample source:Galea | Experimental Variable:d3 Sample source:Galea | Experimental Variable:d3 Sample source:Galea | Experimental Variable:d3 Sample source:Galea | Experimental Variable:d3 Sample source:Galea | Experimental Variable:d3 Sample source:Tissue (PJI) | Experimental Variable:d3 Sample source:Tissue (PJI) | Experimental Variable:d3 Sample source:Tissue (PJI) | Experimental Variable:d3 Sample source:Tissue (PJI) | Experimental Variable:d3 Sample source:Tissue (PJI) | Experimental Variable:d3 Sample source:Galea | Experimental Variable:d7 Sample source:Galea | Experimental Variable:d7 Sample source:Galea | Experimental Variable:d7 Sample source:Galea | Experimental Variable:d7 Sample source:Galea | Experimental Variable:d7 Sample source:Tissue (PJI) | Experimental Variable:d7 Sample source:Tissue (PJI) | Experimental Variable:d7 Sample source:Tissue (PJI) | Experimental Variable:d7 Sample source:Tissue (PJI) | Experimental Variable:d7 Sample source:Tissue (PJI) | Experimental Variable:d7 Sample source:Galea | Experimental Variable:d14 Sample source:Galea | Experimental Variable:d14 Sample source:Galea | Experimental Variable:d14 Sample source:Galea | Experimental Variable:d14 Sample source:Galea | Experimental Variable:d14 Sample source:Tissue (PJI) | Experimental Variable:d14 Sample source:Tissue (PJI) | Experimental Variable:d14 Sample source:Tissue (PJI) | Experimental Variable:d14 Sample source:Tissue (PJI) | Experimental Variable:d14 Sample source:Tissue (PJI) | Experimental Variable:d14 (1S,2R,5S)-2-Isopropyl-5-methylcyclohexyl 3-oxobutanoate 3.07E+04 3.39E+04 2.03E+04 1.14E+04 1.89E+04 3.31E+04 3.52E+04 3.36E+04 3.01E+04 1.56E+04 9.88E+03 3.40E+04 9.33E+03 2.29E+04 3.48E+04 2.28E+04 3.00E+04 3.17E+04 2.48E+04 7.21E+04 1.63E+05 3.60E+04 3.13E+04 8.62E+04 8.50E+04 1.68E+04 1.40E+04 3.69E+04 1.96E+04 8.10E+03 (2E)-decenoic acid 2.17E+04 2.05E+04 1.63E+04 7.97E+03 1.80E+04 2.35E+04 2.09E+04 2.31E+04 1.76E+04 1.60E+04 8.43E+03 2.45E+04 9.32E+03 1.73E+04 1.95E+04 1.19E+04 1.25E+04 2.63E+04 2.13E+04 5.44E+04 6.67E+04 1.77E+04 1.45E+04 6.24E+03 6.97E+04 1.53E+04 6.74E+03 2.73E+04 1.73E+04 4.45E+03 Dihydroartemisinin 4.74E+04 5.41E+04 5.49E+04 2.87E+04 2.82E+04 8.73E+04 5.55E+04 9.83E+04 6.08E+04 3.07E+04 2.07E+04 6.12E+04 1.96E+04 3.81E+04 6.31E+04 4.39E+04 6.05E+04 6.23E+04 6.41E+04 1.85E+05 2.21E+05 4.81E+04 9.73E+04 9.67E+04 1.51E+05 4.61E+04 2.26E+04 7.86E+04 5.73E+04 1.74E+04 (5E,9Z)-farnesyl acetone 1.78E+04 1.27E+04 1.13E+04 6.99E+03 8.38E+03 2.33E+04 1.63E+04 1.69E+04 1.63E+04 1.08E+04 4.52E+03 1.13E+04 4.17E+03 1.13E+04 1.29E+04 1.21E+04 9.01E+03 1.41E+04 8.08E+03 2.42E+04 7.10E+04 8.64E+03 1.78E+04 4.07E+04 4.88E+04 1.27E+04 9.86E+03 2.97E+04 1.33E+04 4.88E+03 (R)-3-hydroxybutyrylcarnitine 5.72E+04 4.72E+04 3.23E+04 1.63E+04 5.40E+04 5.26E+04 4.93E+04 5.03E+04 6.06E+04 6.05E+04 2.98E+04 3.07E+04 4.68E+04 2.44E+04 2.88E+04 3.69E+04 2.50E+04 3.36E+04 4.46E+04 3.85E+04 1.90E+04 1.49E+04 2.48E+04 7.13E+02 7.51E+03 7.33E+04 1.76E+04 2.34E+04 7.83E+04 1.24E+04 15,16-DiHODE 4.48E+04 3.51E+04 2.57E+04 1.34E+04 2.92E+04 9.36E+04 4.25E+04 5.09E+04 5.21E+04 3.07E+04 1.62E+04 5.51E+04 1.15E+04 3.76E+04 4.31E+04 2.94E+04 4.93E+04 4.54E+04 4.25E+04 9.70E+04 2.85E+05 4.96E+04 6.22E+03 1.21E+05 1.01E+05 3.96E+04 2.32E+04 5.57E+04 5.14E+04 1.38E+04 1-Linoleoyl-2-Hydroxy-sn-glycero-3-PC 4.27E+04 3.62E+04 6.27E+04 5.60E+04 6.96E+04 3.05E+04 1.54E+04 2.32E+04 2.37E+04 1.28E+04 1.89E+04 3.62E+04 1.32E+04 2.82E+04 2.70E+04 2.52E+04 2.28E+04 1.16E+04 1.56E+04 2.51E+04 1.31E+05 3.64E+04 3.98E+04 2.11E+05 6.10E+04 5.24E+04 1.02E+04 1.90E+04 3.52E+04 1.13E+04 1-Methylhistamine 1.62E+05 1.64E+05 4.57E+03 4.70E+04 4.69E+03 2.93E+05 2.52E+05 3.12E+05 3.15E+05 2.97E+05 1.31E+05 1.53E+05 9.29E+04 1.90E+05 1.33E+05 1.71E+05 2.03E+05 1.52E+05 2.31E+05 2.38E+05 1.40E+05 1.12E+05 1.47E+05 4.20E+03 9.27E+04 2.18E+05 1.22E+05 3.01E+05 3.55E+05 8.48E+04 1-Methyl-N-(9-methyl-9-azabicyclo[3.3.1]non-3-yl)-1H-indazole-3-carboxamide 3.92E+04 4.19E+04 2.90E+04 1.48E+04 2.51E+04 7.12E+04 4.37E+04 4.70E+04 5.35E+04 2.53E+04 1.67E+04 4.39E+04 1.40E+04 6.28E+04 4.43E+04 3.51E+04 5.22E+04 5.79E+04 4.66E+04 1.23E+05 1.87E+05 3.39E+04 4.79E+04 7.83E+04 1.13E+05 3.98E+04 2.82E+04 6.70E+04 4.61E+04 1.24E+04 1-Stearoyl-2-hydroxy-sn-glycero-3-PE 1.10E+04 9.25E+03 7.80E+03 6.20E+03 8.96E+03 6.31E+03 3.60E+03 4.41E+03 5.98E+03 3.97E+03 4.75E+03 7.33E+03 3.63E+03 1.37E+04 6.10E+03 4.30E+03 4.38E+03 5.09E+03 3.39E+03 3.70E+03 1.78E+04 6.53E+03 7.65E+03 1.70E+04 8.00E+03 9.72E+03 2.45E+03 7.40E+03 4.66E+03 2.14E+03 2-(Diethylamino)ethanol 3.65E+05 6.54E+05 9.36E+05 9.77E+05 1.78E+06 6.20E+06 1.39E+06 1.16E+05 6.02E+06 5.02E+04 2.42E+04 5.75E+04 8.28E+03 9.34E+04 3.16E+06 3.04E+04 5.49E+04 6.68E+04 4.60E+04 1.53E+05 3.13E+05 3.80E+06 1.69E+06 5.26E+06 5.82E+06 5.15E+04 3.08E+04 1.43E+05 6.71E+05 8.26E+03 2,4-dihydroxyheptadec-16-ynyl acetate 3.23E+04 3.06E+04 2.16E+04 1.12E+04 2.41E+04 4.49E+04 2.78E+04 4.16E+04 4.64E+04 2.45E+04 1.13E+04 3.17E+04 1.25E+04 3.25E+04 3.75E+04 2.56E+04 2.71E+04 4.28E+04 4.31E+04 1.15E+05 1.34E+05 3.08E+04 3.90E+04 4.98E+04 1.02E+05 3.29E+04 2.11E+04 4.72E+04 3.47E+04 1.24E+04 2-[4-(3-Hydroxypropyl)-2-methoxyphenoxy]-1,3-propanediol 1.58E+04 2.93E+04 2.24E+04 1.39E+04 1.62E+04 4.24E+04 2.72E+04 3.87E+04 3.50E+04 1.95E+04 9.45E+03 2.65E+04 8.44E+03 2.08E+04 2.64E+04 2.73E+04 5.05E+04 4.01E+04 2.65E+04 7.28E+04 1.18E+05 3.45E+04 3.78E+04 5.98E+04 6.20E+04 2.08E+04 1.45E+04 4.47E+04 2.17E+04 1.22E+04 2'-Deoxycytidine 7.56E+03 1.95E+02 6.26E+03 4.19E+03 2.88E+03 3.96E+03 2.84E+03 1.03E+04 6.98E+03 1.64E+04 1.26E+04 3.15E+04 1.26E+04 4.46E+02 3.36E+03 2.61E+04 4.53E+04 2.66E+04 3.91E+04 2.00E+04 1.98E+04 4.20E+03 6.33E+03 8.65E+02 1.84E+03 7.34E+04 3.08E+04 2.96E+04 1.30E+04 9.21E+03 2-Methoxypyrazine 1.81E+05 1.48E+05 7.07E+04 4.92E+04 5.13E+04 1.95E+05 1.06E+05 1.52E+05 1.75E+05 1.37E+05 3.67E+04 9.61E+04 2.70E+04 2.73E+05 5.46E+04 9.30E+04 1.28E+05 1.09E+05 1.10E+05 9.25E+04 1.63E+05 6.18E+04 6.20E+04 8.72E+03 1.32E+05 8.86E+04 3.24E+04 2.04E+05 1.14E+05 2.22E+04 2-Octenoic acid 3.97E+03 1.10E+04 7.68E+03 6.16E+03 5.68E+03 1.49E+04 9.86E+03 1.13E+04 1.46E+04 5.62E+03 4.38E+03 1.82E+04 2.90E+03 1.60E+04 1.15E+04 1.16E+04 1.45E+04 1.03E+04 1.34E+04 3.62E+04 5.96E+04 9.64E+03 1.28E+04 1.53E+04 4.80E+04 1.15E+04 3.89E+03 2.19E+04 1.66E+04 3.58E+03 3,4-Dihydroxyphenylglycol 5.09E+03 4.75E+03 3.64E+03 2.03E+03 2.28E+03 7.11E+03 4.34E+03 4.57E+03 4.82E+03 2.47E+03 2.19E+03 6.19E+03 1.96E+03 2.26E+03 4.10E+03 2.78E+03 3.86E+03 4.75E+03 4.73E+03 1.28E+04 2.02E+04 4.89E+03 2.72E+03 4.33E+03 6.02E+03 4.23E+03 2.65E+03 4.55E+03 6.00E+03 2.00E+03 3,4-Dimethyl-5-pentyl-2-furanpropanoic acid 1.10E+05 8.55E+04 6.64E+04 3.39E+04 6.44E+04 1.49E+05 8.05E+04 1.12E+05 8.67E+04 5.99E+04 3.35E+04 1.28E+05 2.59E+04 7.02E+04 7.19E+04 5.97E+04 8.86E+04 9.07E+04 8.12E+04 2.07E+05 3.75E+05 8.42E+04 9.51E+04 1.79E+05 2.23E+05 6.58E+04 5.18E+04 1.15E+05 9.87E+04 3.29E+04 3-[(3-Methylbutyl)nitrosoamino]-2-butanone 4.57E+03 5.85E+03 7.60E+03 4.55E+03 6.81E+03 2.71E+04 5.98E+03 5.30E+03 1.24E+04 3.96E+03 8.93E+03 1.63E+04 3.13E+03 3.26E+04 1.26E+04 1.79E+04 1.17E+04 1.63E+04 2.52E+04 5.31E+04 2.01E+05 3.02E+04 1.56E+04 8.41E+04 1.19E+05 1.22E+04 9.56E+03 4.67E+04 1.81E+04 5.10E+03 4-[(2-Isopropyl-5-methylcyclohexyl)oxy]-4-oxobutanoic acid 6.26E+04 3.79E+04 2.42E+04 1.64E+04 2.29E+04 4.26E+04 3.38E+04 4.85E+04 4.32E+04 2.71E+04 1.24E+04 3.64E+04 1.11E+04 3.18E+04 4.45E+04 2.47E+04 4.22E+04 3.78E+04 3.46E+04 1.03E+05 1.01E+05 2.62E+04 3.38E+04 6.39E+04 1.16E+05 2.89E+04 1.68E+04 4.29E+04 3.93E+04 1.29E+04 4-Aminobenzoic acid 7.66E+03 9.22E+03 5.54E+03 9.05E+03 8.18E+03 5.90E+03 4.82E+03 3.95E+03 5.39E+03 2.82E+03 3.69E+03 3.86E+03 4.95E+03 5.13E+03 4.35E+03 3.97E+03 4.72E+03 3.36E+03 4.83E+03 5.26E+03 1.75E+03 2.70E+03 4.85E+03 2.80E+03 5.42E+03 7.82E+03 2.23E+03 6.59E+03 1.24E+04 1.89E+03 4-Hydroxy-5-methoxy-4-[2-methyl-3-(3-methyl-2-buten-1-yl)-2-oxiranyl]-1-oxaspiro[2.5]octan-6-one 4.89E+04 4.19E+04 3.03E+04 1.40E+04 2.61E+04 9.92E+04 4.98E+04 5.43E+04 5.48E+04 2.53E+04 1.25E+04 5.82E+04 1.57E+04 4.40E+04 5.10E+04 4.71E+04 4.33E+04 5.72E+04 3.47E+04 1.40E+05 1.99E+05 4.38E+04 4.72E+04 1.00E+05 1.47E+05 4.31E+04 2.36E+04 6.44E+04 3.88E+04 1.79E+04 5,5'-[Disulfanediylbis({3-[(carboxymethyl)amino]-3-oxo-1,2-propanediyl}imino)]bis(2-amino-5-oxopentanoic acid) 1.13E+04 1.37E+04 1.98E+04 2.62E+04 1.93E+04 3.57E+04 3.76E+04 4.76E+04 5.25E+04 7.67E+04 2.42E+04 2.15E+04 5.30E+04 3.86E+04 2.21E+04 4.09E+04 4.91E+04 4.10E+04 4.70E+04 4.27E+04 1.97E+04 1.47E+04 2.52E+04 1.41E+03 9.38E+03 1.09E+05 2.25E+04 5.53E+04 9.41E+04 1.72E+04 5-Isobutyl-2-oxotetrahydro-3-furancarboxylic acid 1.07E+04 1.18E+04 8.20E+03 3.88E+03 6.51E+03 1.91E+04 1.13E+04 1.64E+04 1.08E+04 2.44E+03 1.98E+03 1.85E+04 1.80E+03 1.09E+04 9.82E+03 1.52E+04 1.52E+04 1.09E+04 1.45E+04 2.53E+04 1.22E+05 1.20E+04 1.88E+04 2.60E+04 4.54E+04 1.10E+04 5.31E+03 1.96E+04 1.82E+04 4.33E+03 6-Methoxyelemicin 8.74E+04 6.11E+04 2.99E+04 2.97E+04 5.63E+04 1.04E+05 1.26E+05 9.39E+04 7.63E+04 3.04E+04 2.56E+04 9.99E+04 2.13E+04 5.80E+04 6.70E+04 5.88E+04 8.49E+04 8.58E+04 8.38E+04 2.10E+05 2.56E+05 6.66E+04 9.91E+04 1.22E+05 2.40E+05 5.15E+04 3.03E+04 8.92E+04 6.28E+04 2.61E+04 7-Hydroxymyoporone 1.45E+05 7.23E+04 6.74E+04 4.03E+04 4.96E+04 1.23E+05 7.96E+04 1.04E+05 1.18E+05 5.61E+04 2.39E+04 8.83E+04 3.27E+04 6.68E+04 7.69E+04 1.01E+05 8.99E+04 1.03E+05 1.07E+05 2.53E+05 3.87E+05 7.31E+04 1.15E+05 1.67E+05 1.77E+05 8.58E+04 4.23E+04 1.31E+05 8.63E+04 3.00E+04 Acetylcarnitine 2.59E+06 2.74E+06 1.19E+06 1.02E+06 5.78E+05 4.23E+06 3.58E+06 4.36E+06 3.76E+06 4.26E+06 1.79E+06 2.41E+06 2.16E+06 2.09E+06 1.94E+06 3.18E+06 3.48E+06 3.17E+06 3.40E+06 3.83E+06 1.84E+06 1.07E+06 1.81E+06 2.50E+04 1.33E+06 5.36E+06 1.63E+06 3.40E+06 6.90E+06 1.25E+06 Adenosine monophosphate 3.32E+04 2.66E+04 2.24E+04 2.59E+04 1.94E+04 2.44E+04 2.11E+04 2.37E+04 2.54E+04 3.80E+04 2.19E+04 2.41E+04 3.32E+04 3.37E+04 1.88E+04 2.32E+04 2.24E+04 2.83E+04 2.82E+04 2.53E+04 2.71E+04 1.13E+04 2.15E+04 7.88E+03 2.15E+04 5.17E+04 1.46E+04 2.99E+04 4.81E+04 7.41E+03 Alpha-GPC 2.90E+04 2.95E+04 1.67E+04 2.89E+04 1.99E+04 1.60E+04 1.20E+04 1.79E+04 1.42E+04 1.28E+04 1.78E+04 2.52E+04 1.85E+04 1.97E+04 2.18E+04 2.38E+04 1.97E+04 4.43E+04 2.60E+04 1.91E+04 6.37E+04 3.02E+04 3.20E+04 1.49E+04 3.37E+04 3.03E+04 8.56E+03 1.96E+04 3.11E+04 6.73E+03 Aminolevulinic acid 1.37E+03 2.87E+03 4.13E+03 3.28E+03 5.02E+03 2.14E+03 1.52E+03 1.48E+03 2.22E+03 1.59E+03 5.44E+02 4.89E+02 1.20E+03 7.74E+02 1.11E+03 1.08E+03 6.86E+02 5.22E+02 1.19E+03 1.06E+03 1.23E+03 1.10E+03 7.55E+02 2.63E+03 5.74E+02 2.39E+03 4.37E+02 9.55E+02 2.60E+03 4.76E+02 Butenylcarnitine 2.63E+04 3.06E+04 2.93E+04 7.58E+03 1.35E+04 7.23E+04 3.88E+04 4.46E+04 4.36E+04 2.09E+04 8.54E+03 4.54E+04 9.43E+03 1.81E+04 2.24E+04 1.90E+04 7.70E+04 2.82E+04 3.75E+04 5.92E+04 2.72E+05 3.78E+04 2.54E+04 1.08E+05 4.45E+04 2.18E+04 8.33E+03 2.78E+04 3.08E+04 2.13E+04 Butylparaben 6.79E+04 4.57E+04 2.51E+04 2.05E+04 3.14E+04 4.91E+04 3.71E+04 5.16E+04 4.91E+04 2.53E+04 1.70E+04 6.11E+04 1.40E+04 4.13E+04 4.83E+04 4.31E+04 4.32E+04 6.15E+04 4.69E+04 1.28E+05 2.79E+05 4.53E+04 4.81E+04 9.57E+04 1.27E+05 3.20E+04 2.03E+04 7.42E+04 4.76E+04 1.71E+04 C14-Carnitine 2.01E+04 2.59E+04 1.87E+04 9.91E+03 1.18E+04 3.02E+05 7.06E+04 4.67E+04 9.30E+04 7.48E+04 7.51E+03 2.87E+04 1.66E+04 3.18E+04 1.28E+04 2.34E+04 3.48E+04 2.82E+04 4.53E+04 1.16E+05 1.67E+04 1.01E+04 2.73E+04 5.18E+03 4.20E+04 7.26E+04 2.22E+04 7.07E+04 7.59E+04 5.76E+03 C4-Carnitine 5.10E+04 5.85E+04 3.02E+04 3.72E+04 3.96E+04 6.31E+04 6.16E+04 5.60E+04 7.50E+04 6.86E+04 7.76E+03 1.23E+04 2.63E+04 2.72E+04 2.29E+04 2.42E+04 4.54E+04 7.34E+04 1.71E+04 4.19E+04 3.20E+04 1.50E+04 2.01E+04 5.33E+03 2.96E+04 6.13E+04 1.75E+04 1.68E+04 8.09E+04 1.50E+04 cis-3-Hexenyl Pyruvate 5.97E+04 4.69E+04 2.74E+04 7.45E+03 3.95E+04 4.70E+04 3.11E+04 4.86E+04 4.41E+04 2.34E+04 2.42E+04 7.45E+04 1.52E+04 4.78E+04 5.39E+04 3.72E+04 8.89E+04 5.84E+04 4.48E+04 1.03E+05 2.40E+05 3.55E+04 5.17E+04 9.66E+04 1.24E+05 2.41E+04 2.04E+04 6.53E+04 3.35E+04 1.69E+04 Colfosceril palmitate 2.68E+05 4.33E+05 1.07E+05 1.20E+04 7.27E+04 6.57E+04 5.96E+04 1.05E+05 1.20E+05 7.30E+04 5.03E+04 2.11E+04 2.83E+04 1.13E+04 1.13E+05 1.07E+05 5.08E+04 1.59E+05 1.04E+05 1.80E+05 8.62E+04 2.91E+04 2.69E+04 1.18E+03 1.58E+04 1.02E+05 5.07E+04 1.44E+04 2.57E+04 4.81E+03 Cyclohexylamine 2.52E+05 1.02E+05 1.53E+05 1.57E+05 2.65E+05 1.02E+06 6.50E+05 4.47E+05 7.11E+05 6.19E+05 5.73E+04 2.05E+05 1.04E+05 6.34E+05 5.11E+05 2.74E+05 2.28E+04 2.12E+04 2.20E+05 4.89E+04 4.11E+06 4.82E+05 2.47E+05 8.69E+05 3.76E+05 2.32E+04 1.19E+04 4.78E+05 2.39E+05 8.70E+04 Cytosine 1.11E+04 7.76E+03 5.63E+03 2.18E+03 2.71E+03 5.73E+03 4.86E+03 1.04E+04 6.54E+03 1.47E+04 1.32E+04 3.28E+04 1.22E+04 5.69E+04 4.20E+03 2.62E+04 4.64E+04 3.23E+04 3.74E+04 2.30E+04 2.54E+04 5.54E+03 9.96E+02 8.03E+02 6.37E+03 6.88E+04 2.94E+04 3.47E+04 1.21E+04 8.38E+03 Deanol 2.84E+05 1.72E+05 1.20E+04 1.17E+04 1.64E+04 1.31E+06 3.81E+05 3.43E+05 3.11E+05 2.25E+05 1.54E+05 5.50E+05 7.10E+04 3.60E+05 2.81E+05 4.15E+05 2.63E+05 2.55E+05 4.44E+05 5.95E+05 2.80E+06 4.48E+05 2.32E+05 4.59E+04 2.28E+05 2.49E+05 1.87E+05 4.14E+05 2.44E+05 1.07E+05 Diethylamine 9.98E+04 8.62E+04 6.72E+04 5.32E+04 7.98E+04 2.23E+05 1.20E+05 1.27E+05 1.47E+05 8.07E+04 1.71E+04 3.96E+04 2.77E+04 1.13E+05 1.06E+05 2.34E+04 5.22E+04 6.28E+04 3.45E+04 1.51E+05 3.54E+05 7.74E+04 1.17E+05 3.09E+05 3.83E+05 3.77E+04 1.89E+04 1.54E+05 8.87E+04 1.27E+04 DL-Citrulline 2.40E+04 1.89E+04 2.13E+04 2.14E+04 1.33E+04 2.54E+04 1.53E+04 1.99E+04 2.21E+04 2.34E+04 1.35E+04 1.13E+04 1.25E+04 1.28E+04 8.44E+03 1.99E+04 1.64E+04 1.04E+04 1.26E+04 9.08E+03 1.79E+04 9.66E+03 1.20E+04 2.38E+03 9.84E+03 2.84E+04 1.06E+04 1.93E+04 3.25E+04 4.74E+03 DL-Glutamic acid 1.48E+05 1.39E+05 1.14E+05 1.39E+05 7.87E+04 1.07E+05 8.61E+04 1.02E+05 1.23E+05 1.26E+05 7.51E+04 8.87E+04 7.53E+04 7.18E+04 5.42E+04 6.68E+04 5.43E+04 7.87E+04 5.62E+04 5.50E+04 9.66E+04 3.67E+04 6.76E+04 1.22E+04 4.50E+04 1.55E+05 2.63E+04 4.52E+04 1.07E+05 1.41E+04 D-PANTOTHENIC ACID 5.32E+03 8.42E+03 1.07E+04 1.27E+04 9.04E+03 3.05E+03 2.12E+03 1.30E+03 3.79E+03 1.70E+03 1.16E+03 1.77E+03 2.86E+03 2.85E+03 3.79E+03 1.45E+03 6.37E+02 3.25E+02 1.48E+03 1.17E+03 1.03E+04 2.81E+03 3.69E+03 1.09E+03 5.50E+03 8.21E+03 1.71E+03 4.06E+03 5.14E+03 5.36E+02 Ethylenediaminetetraacetic acid 2.44E+05 1.56E+05 5.85E+05 3.35E+05 5.53E+05 1.84E+05 9.65E+04 1.12E+05 9.01E+04 4.87E+04 3.12E+04 4.06E+04 2.32E+04 1.47E+04 5.86E+04 2.05E+04 1.52E+04 6.31E+03 1.41E+04 2.14E+04 2.64E+05 3.33E+04 6.02E+04 5.55E+05 4.75E+05 1.69E+04 3.93E+03 3.41E+03 2.07E+04 5.49E+03 g-Butyrobetaine 6.14E+04 3.78E+04 2.50E+04 4.44E+04 2.96E+04 5.43E+04 3.28E+04 3.57E+04 4.79E+04 4.91E+04 2.44E+04 2.37E+04 2.84E+04 3.45E+04 1.65E+04 3.05E+04 3.88E+04 2.88E+04 3.79E+04 4.31E+04 2.63E+04 1.75E+04 2.20E+04 2.82E+03 1.46E+04 6.47E+04 1.82E+04 3.50E+04 6.60E+04 1.21E+04 HEPES 3.50E+05 3.81E+06 1.22E+06 1.59E+04 3.40E+05 3.25E+03 1.47E+04 1.16E+04 2.25E+03 1.58E+02 9.30E+04 4.32E+05 1.26E+05 3.41E+05 4.02E+05 1.74E+05 2.89E+04 6.21E+04 5.26E+03 1.38E+03 1.15E+06 3.45E+05 5.26E+05 3.74E+02 1.46E+06 2.00E+04 8.00E+02 1.94E+04 4.09E+04 1.38E+04 hercynine 1.40E+04 1.59E+04 9.62E+03 5.29E+03 4.82E+03 1.63E+04 1.16E+04 1.64E+04 1.43E+04 1.28E+04 1.20E+04 1.28E+04 1.03E+04 1.70E+04 1.02E+04 1.69E+04 1.97E+04 1.47E+04 1.82E+04 2.36E+04 1.22E+04 6.42E+03 1.06E+04 1.71E+03 9.07E+03 1.80E+04 9.76E+03 3.44E+04 2.33E+04 5.43E+03 Hexylresorcinol 1.20E+05 1.00E+05 9.16E+04 3.80E+04 6.94E+04 1.40E+05 1.20E+05 1.42E+05 1.22E+05 6.96E+04 4.44E+04 1.39E+05 3.31E+04 1.16E+05 1.01E+05 7.79E+04 1.17E+05 1.47E+05 1.39E+05 2.60E+05 5.56E+05 7.68E+04 1.29E+05 2.01E+05 2.73E+05 1.03E+05 6.23E+04 1.50E+05 1.00E+05 3.29E+04 His-pro 1.60E+03 1.81E+03 1.80E+03 1.82E+03 1.41E+03 5.97E+03 1.46E+03 1.12E+03 2.55E+03 2.80E+02 2.21E+02 7.14E+02 3.02E+02 3.38E+02 1.42E+03 1.01E+03 7.63E+02 4.66E+02 5.29E+02 2.40E+03 3.72E+03 1.10E+03 1.21E+03 1.13E+03 7.59E+03 2.04E+03 4.98E+02 6.59E+02 1.65E+03 3.36E+02 Histamine 1.06E+06 9.02E+05 3.27E+05 2.52E+05 8.57E+04 6.53E+05 5.80E+05 7.13E+05 8.21E+05 8.54E+05 8.38E+05 6.97E+05 4.06E+05 1.21E+06 3.64E+05 5.82E+05 7.49E+05 6.42E+05 4.03E+05 3.10E+05 2.52E+05 1.80E+05 2.29E+05 7.98E+03 7.35E+04 9.50E+05 2.50E+05 9.84E+05 6.12E+05 1.05E+05 Inosinic acid 3.40E+03 2.79E+03 2.63E+04 2.27E+04 3.23E+04 2.15E+04 2.16E+04 2.54E+04 2.85E+04 1.38E+04 1.41E+04 5.53E+04 1.79E+04 5.48E+04 6.71E+04 3.31E+04 3.16E+04 1.04E+04 2.24E+04 3.98E+04 2.31E+05 6.26E+04 8.78E+04 1.43E+05 4.11E+05 1.92E+04 8.19E+03 1.75E+04 2.51E+04 7.02E+03 Inspra 4.60E+05 3.09E+05 1.99E+05 9.08E+04 2.33E+05 7.74E+05 1.39E+05 4.39E+05 6.31E+05 3.13E+05 3.55E+05 3.06E+05 2.17E+05 1.10E+06 4.51E+05 2.86E+05 3.43E+05 7.95E+05 7.47E+05 1.82E+06 1.68E+06 2.89E+05 4.02E+05 3.78E+05 6.41E+05 2.11E+05 2.25E+05 5.17E+05 1.41E+06 7.00E+04 L-(-)-Serine 2.83E+04 3.22E+04 2.60E+04 2.36E+04 1.63E+04 4.75E+04 2.64E+04 2.86E+04 4.20E+04 3.65E+04 2.47E+04 2.38E+04 2.81E+04 2.04E+04 2.00E+04 2.64E+04 2.60E+04 1.65E+04 2.55E+04 1.64E+04 2.62E+04 1.45E+04 2.14E+04 5.42E+03 1.79E+04 4.86E+04 1.44E+04 2.36E+04 5.99E+04 9.57E+03 L-(+)-Arginine 6.02E+04 1.60E+05 1.21E+05 1.28E+05 1.11E+05 7.16E+04 4.62E+04 5.04E+04 6.15E+04 6.05E+04 4.72E+04 5.60E+04 5.29E+04 3.98E+04 4.55E+04 4.80E+04 3.69E+04 3.05E+04 3.85E+04 3.60E+04 1.23E+05 4.97E+04 7.12E+04 5.96E+04 1.04E+05 8.46E+04 2.56E+04 5.36E+04 8.83E+04 1.67E+04 L-(+)-Aspartic acid 4.91E+04 4.74E+04 2.70E+04 3.01E+04 1.70E+04 3.78E+04 3.05E+04 3.69E+04 3.31E+04 3.84E+04 2.09E+04 2.62E+04 2.22E+04 2.06E+04 1.45E+04 2.75E+04 2.21E+04 3.67E+04 2.14E+04 1.95E+04 3.32E+04 1.52E+04 2.54E+04 4.90E+03 1.66E+04 5.20E+04 1.16E+04 2.08E+04 4.20E+04 7.12E+03 L-(+)-ERGOTHIONEINE 5.23E+04 7.77E+04 2.64E+03 6.74E+02 5.28E+02 1.64E+05 5.56E+04 1.64E+05 1.85E+05 1.80E+05 1.01E+05 1.09E+05 1.92E+05 1.89E+04 1.06E+05 1.97E+05 7.56E+04 4.50E+04 1.43E+05 1.48E+05 7.41E+04 8.16E+04 1.50E+05 6.81E+02 5.29E+04 4.64E+05 1.12E+05 1.03E+05 4.91E+05 9.59E+04 L-(+)-glutamine 5.02E+04 1.07E+05 5.94E+04 5.91E+04 3.11E+04 3.91E+04 3.42E+04 3.94E+04 5.40E+04 5.81E+04 4.27E+04 4.25E+04 4.19E+04 4.32E+04 3.43E+04 4.16E+04 3.43E+04 3.05E+04 2.86E+04 1.58E+04 8.35E+04 2.43E+04 3.51E+04 2.50E+04 4.46E+04 7.05E+04 1.88E+04 3.99E+04 5.76E+04 1.30E+04 L-(+)-Leucine 2.17E+05 4.21E+05 2.43E+05 3.64E+05 1.89E+05 2.63E+05 1.84E+05 2.22E+05 3.50E+05 3.41E+05 2.10E+05 2.00E+05 2.29E+05 2.20E+05 1.94E+05 2.31E+05 2.10E+05 1.85E+05 2.13E+05 1.62E+05 3.16E+05 1.58E+05 1.73E+05 1.08E+05 2.09E+05 4.28E+05 1.23E+05 1.44E+05 4.94E+05 8.26E+04 L-(+)-Valine 1.54E+06 2.90E+06 1.95E+06 2.07E+06 1.86E+06 2.93E+06 2.00E+06 2.67E+06 2.65E+06 2.08E+06 1.52E+06 3.33E+06 1.24E+06 2.05E+06 2.01E+06 2.19E+06 3.01E+06 2.21E+06 2.94E+06 4.54E+06 3.51E+06 2.22E+06 4.26E+06 1.29E+06 1.78E+06 3.64E+06 1.65E+06 2.94E+06 4.27E+06 8.16E+05 L-Palmitoylcarnitine 3.09E+05 4.58E+05 3.39E+05 2.16E+05 1.90E+05 4.76E+05 1.24E+05 4.37E+05 4.42E+05 3.06E+05 3.04E+05 1.03E+05 1.74E+05 2.61E+05 1.69E+05 3.89E+05 8.22E+04 4.37E+05 3.58E+05 2.33E+05 6.51E+04 3.58E+04 1.74E+05 2.76E+04 1.27E+05 4.86E+05 1.35E+05 3.63E+05 2.70E+05 2.86E+04 L-Proline 2.99E+05 3.99E+05 3.47E+05 1.57E+05 1.37E+05 3.17E+05 2.51E+05 2.82E+05 4.09E+05 3.88E+05 2.34E+05 2.06E+05 2.89E+05 2.53E+05 1.97E+05 2.62E+05 2.48E+05 1.89E+05 2.12E+05 1.45E+05 1.54E+05 1.23E+05 1.48E+05 2.07E+04 9.68E+04 4.91E+05 1.21E+05 2.32E+05 4.38E+05 7.73E+04 LQ5400000 7.18E+03 4.95E+03 3.32E+03 4.22E+03 3.41E+03 7.04E+03 4.10E+03 6.12E+03 8.36E+03 2.63E+03 2.03E+03 6.62E+03 1.89E+03 3.55E+03 4.94E+03 4.19E+03 3.48E+03 4.88E+03 5.25E+03 1.50E+04 3.57E+04 4.11E+03 9.45E+03 4.98E+03 1.25E+04 1.95E+03 3.69E+03 1.49E+04 5.45E+03 1.55E+03 lys-leu 1.04E+04 1.21E+04 1.26E+04 1.13E+04 9.61E+03 1.47E+04 1.25E+04 1.65E+04 1.94E+04 1.94E+04 1.01E+04 8.31E+03 1.08E+04 8.20E+03 6.15E+03 1.03E+04 1.04E+04 6.98E+03 9.21E+03 1.15E+04 6.76E+03 4.88E+03 9.14E+03 1.77E+03 4.26E+03 1.78E+04 5.39E+03 9.85E+03 1.81E+04 3.67E+03 Methyl Jasmonate 7.63E+04 6.61E+04 5.39E+04 2.96E+04 4.42E+04 1.01E+05 5.81E+04 8.33E+04 9.61E+04 4.06E+04 2.66E+04 1.13E+05 1.44E+04 6.66E+04 7.08E+04 6.33E+04 6.08E+04 9.48E+04 8.58E+04 2.16E+05 2.23E+05 5.07E+04 7.96E+04 1.34E+05 1.68E+05 7.09E+04 4.80E+04 1.24E+05 5.95E+04 2.04E+04 MFCD00036904 3.34E+05 4.31E+05 2.85E+05 3.24E+05 2.00E+05 8.91E+04 4.46E+04 9.39E+04 1.27E+05 8.57E+04 1.20E+05 8.17E+04 1.38E+05 2.05E+04 2.77E+05 1.04E+05 5.32E+04 2.49E+05 8.71E+04 1.74E+05 1.85E+05 6.43E+04 4.95E+04 6.93E+04 1.72E+05 2.54E+05 5.60E+04 3.64E+04 1.90E+04 9.86E+03 MFCD00133435 3.32E+04 5.47E+04 3.61E+04 4.39E+04 5.72E+04 3.19E+04 1.52E+04 2.70E+04 2.22E+04 1.35E+04 3.34E+04 1.40E+04 1.74E+04 6.16E+04 3.52E+04 1.68E+04 3.37E+04 1.96E+04 2.44E+04 3.32E+04 2.16E+05 1.49E+04 2.34E+04 5.52E+04 1.80E+05 2.54E+04 1.78E+04 4.44E+04 3.87E+04 9.94E+03 MFCD02259286 4.26E+04 8.77E+04 5.61E+04 1.75E+04 2.22E+04 3.54E+05 1.63E+05 1.61E+05 2.07E+05 1.17E+05 4.55E+04 1.60E+05 3.13E+04 3.24E+05 2.95E+04 8.23E+04 1.74E+05 1.17E+05 2.38E+05 1.86E+05 3.29E+05 1.06E+05 5.96E+04 1.14E+04 5.98E+05 2.26E+05 5.94E+04 2.85E+05 1.88E+05 4.52E+04 MFCD09953737 1.27E+03 1.40E+03 1.21E+03 1.97E+05 6.79E+02 1.21E+03 1.46E+03 9.64E+02 1.83E+03 7.10E+02 4.83E+02 1.53E+03 4.75E+02 7.11E+02 1.40E+03 6.67E+02 5.79E+02 6.83E+02 7.98E+02 7.98E+03 4.90E+03 1.03E+03 1.20E+03 2.48E+03 1.94E+03 1.37E+03 5.98E+02 1.10E+03 2.87E+03 6.00E+02 N(1)-acetylspermidine 1.92E+05 2.01E+05 2.09E+05 2.41E+05 1.89E+05 2.49E+05 1.80E+05 1.54E+05 2.83E+05 2.66E+05 8.57E+04 1.18E+05 6.29E+04 1.20E+05 1.08E+05 1.05E+05 7.46E+04 4.85E+04 1.02E+05 6.23E+04 7.39E+04 5.40E+04 7.10E+04 1.70E+04 5.05E+04 1.71E+05 4.36E+04 7.05E+04 1.60E+05 3.35E+04 N-[(15Z)-tetracosenoyl]sphing-4-enine-1-phosphocholine 8.59E+04 7.37E+04 3.50E+04 1.46E+04 2.13E+04 1.08E+05 3.45E+04 1.24E+05 1.56E+05 9.29E+04 4.01E+04 3.78E+04 2.51E+04 8.66E+04 4.50E+04 8.68E+04 5.47E+04 1.15E+05 1.16E+05 1.92E+05 9.29E+04 3.17E+04 6.52E+04 1.78E+03 7.74E+04 1.74E+05 5.88E+04 4.90E+04 1.19E+05 1.46E+04 N-{3-[(4-Acetamidobutyl)amino]propyl}acetamide 1.45E+04 1.51E+04 3.88E+03 2.17E+02 1.41E+02 1.98E+04 1.13E+04 9.09E+03 1.85E+04 1.81E+04 1.12E+04 9.90E+03 7.59E+03 7.20E+03 6.73E+03 6.57E+03 4.12E+03 2.49E+03 4.92E+03 2.32E+03 1.50E+03 1.94E+03 2.00E+03 6.57E+02 1.05E+03 1.53E+04 2.12E+03 5.24E+03 1.15E+04 2.20E+03 PC 2.00E+05 2.10E+05 9.81E+04 5.00E+04 6.92E+04 1.42E+05 9.08E+04 1.92E+05 2.29E+05 1.73E+05 9.12E+04 8.01E+04 4.67E+04 3.35E+04 2.63E+05 1.71E+05 9.70E+04 2.30E+05 1.74E+05 2.56E+05 1.15E+05 1.80E+04 6.00E+04 7.51E+02 1.52E+05 1.83E+05 7.74E+04 2.51E+05 9.16E+04 5.46E+04 PEG-4 2.28E+05 2.80E+05 1.34E+05 7.67E+04 1.38E+05 4.03E+05 1.31E+05 2.46E+05 1.66E+05 1.36E+05 6.48E+04 3.66E+05 4.37E+04 1.65E+05 1.95E+05 1.28E+05 1.66E+05 2.09E+05 2.26E+05 6.64E+05 1.03E+06 1.19E+05 2.13E+05 4.38E+05 1.45E+06 1.90E+05 7.91E+04 2.61E+05 1.48E+05 8.34E+04 Platelet-activating factor 1.91E+05 2.51E+05 1.09E+05 9.46E+04 9.05E+04 3.55E+04 1.22E+04 5.88E+04 5.66E+04 4.54E+04 7.64E+04 2.89E+04 4.23E+04 1.60E+04 3.07E+05 6.38E+04 6.65E+03 7.21E+04 5.47E+04 1.12E+05 4.26E+04 1.11E+04 3.94E+04 5.78E+04 1.36E+05 1.39E+05 2.94E+04 9.06E+03 1.42E+04 4.90E+03 propionylcarnitine 7.25E+04 8.75E+04 6.74E+04 5.53E+04 3.46E+04 1.66E+05 1.25E+05 1.13E+05 8.60E+04 9.41E+04 4.21E+04 5.42E+04 4.83E+04 5.31E+04 3.55E+04 5.87E+04 5.58E+04 7.76E+04 6.90E+04 9.07E+04 3.64E+04 3.91E+04 7.68E+04 1.34E+04 4.88E+04 9.30E+04 3.04E+04 6.68E+04 1.50E+05 2.68E+04 Propylparaben 5.65E+04 3.05E+04 1.88E+04 7.55E+03 2.63E+04 6.29E+04 2.59E+04 3.16E+04 2.79E+04 2.06E+04 1.11E+04 3.48E+04 1.01E+04 2.89E+04 4.12E+04 4.80E+04 3.63E+04 5.00E+04 4.45E+04 8.73E+04 1.68E+05 2.43E+04 5.89E+04 6.08E+04 9.78E+04 2.11E+04 1.53E+04 5.07E+04 2.28E+04 1.04E+04 R79646H5Z8 2.42E+04 2.11E+04 1.34E+04 5.43E+03 1.12E+04 2.95E+04 2.33E+04 1.74E+04 2.44E+04 1.17E+04 9.29E+03 2.50E+04 7.42E+03 2.12E+04 2.25E+04 2.29E+04 2.87E+04 2.55E+04 1.84E+04 7.21E+04 1.24E+05 1.68E+04 2.21E+04 4.00E+04 4.89E+04 1.45E+04 9.98E+03 2.49E+04 2.43E+04 8.72E+03 RG5927200 1.78E+04 1.72E+04 1.46E+04 6.33E+03 1.00E+04 3.59E+04 1.65E+04 1.75E+04 1.85E+04 1.10E+04 7.55E+03 2.51E+04 5.83E+03 1.41E+04 1.70E+04 1.13E+04 1.58E+04 2.02E+04 1.69E+04 6.81E+04 8.72E+04 1.54E+04 2.27E+04 3.45E+04 7.15E+04 1.81E+04 1.05E+04 2.89E+04 1.87E+04 5.69E+03 Taurine 1.36E+06 1.51E+06 6.90E+05 1.46E+06 8.22E+05 2.27E+06 1.67E+06 2.08E+06 2.40E+06 2.30E+06 1.20E+06 1.85E+06 1.46E+06 1.85E+06 1.51E+06 2.21E+06 2.29E+06 1.68E+06 2.13E+06 2.82E+06 1.14E+06 8.06E+05 1.30E+06 2.65E+05 8.78E+05 3.38E+06 9.86E+05 2.23E+06 3.52E+06 6.99E+05 Tetranor 12-HETE 4.76E+04 4.28E+04 3.49E+04 1.52E+04 3.25E+04 7.00E+04 3.97E+04 4.72E+04 5.61E+04 3.17E+04 1.30E+04 5.34E+04 1.46E+04 3.79E+04 4.42E+04 3.23E+04 4.96E+04 5.47E+04 4.52E+04 1.32E+05 2.25E+05 4.42E+04 4.79E+04 8.88E+04 1.21E+05 4.08E+04 2.64E+04 6.16E+04 4.21E+04 1.55E+04 Urocanic acid 1.81E+04 8.56E+03 1.37E+04 1.05E+04 1.84E+04 1.07E+05 2.65E+04 1.84E+04 2.92E+04 5.12E+03 2.18E+03 4.70E+03 2.24E+03 6.37E+03 6.52E+03 1.71E+03 2.20E+03 4.62E+03 1.87E+04 3.71E+03 2.70E+04 4.19E+03 6.99E+03 2.78E+04 4.27E+04 3.78E+04 1.67E+03 7.21E+03 8.54E+04 3.03E+03 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Human Metabolome Database (1S,2R,5S)-2-Isopropyl-5-methylcyclohexyl 3-oxobutanoate HMDB0032369 (2E)-decenoic acid HMDB0010726 Dihydroartemisinin HMDB0060593 (5E,9Z)-farnesyl acetone HMDB0034495 (R)-3-hydroxybutyrylcarnitine HMDB0062735 15,16-DiHODE HMDB0010208 1-Linoleoyl-2-Hydroxy-sn-glycero-3-PC HMDB0061692 1-Methylhistamine HMDB0000898 1-Methyl-N-(9-methyl-9-azabicyclo[3.3.1]non-3-yl)-1H-indazole-3-carboxamide HMDB0015026 1-Stearoyl-2-hydroxy-sn-glycero-3-PE HMDB0011130 2-(Diethylamino)ethanol HMDB0033971 2,4-dihydroxyheptadec-16-ynyl acetate HMDB0031048 2-[4-(3-Hydroxypropyl)-2-methoxyphenoxy]-1,3-propanediol HMDB0040352 2'-Deoxycytidine HMDB0000014 2-Methoxypyrazine HMDB0033156 2-Octenoic acid HMDB0001568 3,4-Dihydroxyphenylglycol HMDB0000318 3,4-Dimethyl-5-pentyl-2-furanpropanoic acid HMDB0112083 3-[(3-Methylbutyl)nitrosoamino]-2-butanone HMDB0033553 4-[(2-Isopropyl-5-methylcyclohexyl)oxy]-4-oxobutanoic acid HMDB0036143 4-Aminobenzoic acid HMDB0001392 4-Hydroxy-5-methoxy-4-[2-methyl-3-(3-methyl-2-buten-1-yl)-2-oxiranyl]-1-oxaspiro[2.5]octan-6-one HMDB0038120 5,5'-[Disulfanediylbis({3-[(carboxymethyl)amino]-3-oxo-1,2-propanediyl}imino)]bis(2-amino-5-oxopentanoic acid) HMDB0003337 5-Isobutyl-2-oxotetrahydro-3-furancarboxylic acid HMDB0030992 6-Methoxyelemicin HMDB0034046 7-Hydroxymyoporone HMDB0035210 Acetylcarnitine HMDB0000201 Adenosine monophosphate HMDB0000045 Alpha-GPC HMDB0000086 Aminolevulinic acid HMDB0001149 Butenylcarnitine HMDB0013126 Butylparaben HMDB0032575 C14-Carnitine HMDB0254979 C4-Carnitine HMDB0002013 cis-3-Hexenyl Pyruvate HMDB0038276 Colfosceril palmitate HMDB0000564 Cyclohexylamine HMDB0031404 Cytosine HMDB0000630 Deanol HMDB0032231 Diethylamine HMDB0041878 DL-Citrulline HMDB0000904 DL-Glutamic acid HMDB0060475 D-PANTOTHENIC ACID HMDB0000210 Ethylenediaminetetraacetic acid HMDB0015109 g-Butyrobetaine HMDB0006831 HEPES HMDB0062295 hercynine HMDB0029422 Hexylresorcinol HMDB0032567 His-pro HMDB0028893 Histamine HMDB0000870 Inosinic acid HMDB0000175 Inspra HMDB0014838 L-(-)-Serine HMDB0000187 L-(+)-Arginine HMDB0000517 L-(+)-Aspartic acid HMDB0000191 L-(+)-ERGOTHIONEINE HMDB0003045 L-(+)-glutamine HMDB0000641 L-(+)-Leucine HMDB0000687 L-(+)-Valine HMDB0000883 L-Palmitoylcarnitine HMDB0000222 L-Proline HMDB0000162 LQ5400000 HMDB0041485 lys-leu HMDB0028955 Methyl Jasmonate HMDB0036583 MFCD00036904 HMDB0010382 MFCD00133435 HMDB0002815 MFCD02259286 HMDB0010168 MFCD09953737 HMDB0000953 N(1)-acetylspermidine HMDB0001276 N-[(15Z)-tetracosenoyl]sphing-4-enine-1-phosphocholine HMDB0011696 N-{3-[(4-Acetamidobutyl)amino]propyl}acetamide HMDB0041947 PC HMDB0062690 PEG-4 HMDB0094708 Platelet-activating factor HMDB0062195 propionylcarnitine HMDB0000824 Propylparaben HMDB0032574 R79646H5Z8 HMDB0240222 RG5927200 HMDB0030428 Taurine HMDB0000251 Tetranor 12-HETE HMDB0060055 Urocanic acid HMDB0000301 METABOLITES_END #END #METABOLOMICS WORKBENCH dhananjay_shinde5_20240914_131224 DATATRACK_ID:5191 STUDY_ID:ST003482 ANALYSIS_ID:AN005718 PROJECT_ID:PR002139 VERSION 1 CREATED_ON September 14, 2024, 3:09 pm #PROJECT PR:PROJECT_TITLE Tissue niche influences immune and metabolic profiles to Staphylococcus aureus PR:PROJECT_TITLE biofilm infection PR:PROJECT_TYPE Intracellular data PR:PROJECT_SUMMARY Infection is a devastating post-surgical complication, often requiring PR:PROJECT_SUMMARY additional procedures and prolonged antibiotic therapy. This is especially PR:PROJECT_SUMMARY relevant for craniotomy and prosthetic joint infections (PJI), both of which are PR:PROJECT_SUMMARY characterized by biofilm formation on the bone or implant surface, respectively, PR:PROJECT_SUMMARY with S. aureus representing a primary cause. The local tissue microenvironment PR:PROJECT_SUMMARY likely has profound effects on immune attributes that can influence treatment PR:PROJECT_SUMMARY efficacy, which becomes critical to consider when developing therapeutics for PR:PROJECT_SUMMARY biofilm infections. However, the extent to which distinct tissue niches PR:PROJECT_SUMMARY influence immune function during biofilm development remains relatively unknown. PR:PROJECT_SUMMARY To address this, we compare the metabolomic, transcriptomic, and functional PR:PROJECT_SUMMARY attributes of leukocytes in mouse models of S. aureus craniotomy and PJI PR:PROJECT_SUMMARY complemented with patient samples from both infection modalities, which reveals PR:PROJECT_SUMMARY profound tissue niche-dependent differences in nucleic acid, amino acid, and PR:PROJECT_SUMMARY lipid metabolism with links to immune modulation. These signatures are both PR:PROJECT_SUMMARY spatially and temporally distinct, differing not only between infection sites PR:PROJECT_SUMMARY but evolving over time within a single model. Collectively, this demonstrates PR:PROJECT_SUMMARY that biofilms elicit unique immune and metabolic responses that are heavily PR:PROJECT_SUMMARY influenced by the local tissue microenvironment, which will likely have PR:PROJECT_SUMMARY important implications when designing therapeutic approaches targeting these PR:PROJECT_SUMMARY infections. PR:INSTITUTE UNMC PR:DEPARTMENT Department of pathology, microbiology and immunology PR:LABORATORY Pathology, Microbiology and Immunology PR:LAST_NAME Dhananjay PR:FIRST_NAME Shinde PR:ADDRESS DRC 2, 7066, UNMC, Emily st PR:EMAIL dhananjay.shinde@unmc.edu PR:PHONE 4025597623 #STUDY ST:STUDY_TITLE Tissue niche influences immune and metabolic profiles to Staphylococcus aureus ST:STUDY_TITLE biofilm infection ST:STUDY_SUMMARY Infection is a devastating post-surgical complication, often requiring ST:STUDY_SUMMARY additional procedures and prolonged antibiotic therapy. This is especially ST:STUDY_SUMMARY relevant for craniotomy and prosthetic joint infections (PJI), both of which are ST:STUDY_SUMMARY characterized by biofilm formation on the bone or implant surface, respectively, ST:STUDY_SUMMARY with S. aureus representing a primary cause. The local tissue microenvironment ST:STUDY_SUMMARY likely has profound effects on immune attributes that can influence treatment ST:STUDY_SUMMARY efficacy, which becomes critical to consider when developing therapeutics for ST:STUDY_SUMMARY biofilm infections. However, the extent to which distinct tissue niches ST:STUDY_SUMMARY influence immune function during biofilm development remains relatively unknown. ST:STUDY_SUMMARY To address this, we compare the metabolomic, transcriptomic, and functional ST:STUDY_SUMMARY attributes of leukocytes in mouse models of S. aureus craniotomy and PJI ST:STUDY_SUMMARY complemented with patient samples from both infection modalities, which reveals ST:STUDY_SUMMARY profound tissue niche-dependent differences in nucleic acid, amino acid, and ST:STUDY_SUMMARY lipid metabolism with links to immune modulation. These signatures are both ST:STUDY_SUMMARY spatially and temporally distinct, differing not only between infection sites ST:STUDY_SUMMARY but evolving over time within a single model. Collectively, this demonstrates ST:STUDY_SUMMARY that biofilms elicit unique immune and metabolic responses that are heavily ST:STUDY_SUMMARY influenced by the local tissue microenvironment, which will likely have ST:STUDY_SUMMARY important implications when designing therapeutic approaches targeting these ST:STUDY_SUMMARY infections. ST:INSTITUTE UNMC ST:DEPARTMENT Department of pathology, microbiology and immunology ST:LAST_NAME Shinde ST:FIRST_NAME Dhananjay ST:ADDRESS DRC 2, 7066, UNMC, Emily st ST:EMAIL dhananjay.shinde@unmc.edu ST:PHONE 4025597623 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:GENDER Male and female #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 - d3-Crain_1 Sample source:Galea | Experimental Variable:d3 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-Crain_1.mzXML SUBJECT_SAMPLE_FACTORS - d3-Crain_2 Sample source:Galea | Experimental Variable:d3 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-Crain_2.mzXML SUBJECT_SAMPLE_FACTORS - d3-Crain_3 Sample source:Galea | Experimental Variable:d3 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-Crain_3.mzXML SUBJECT_SAMPLE_FACTORS - d3-Crain_4 Sample source:Galea | Experimental Variable:d3 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-Crain_4.mzXML SUBJECT_SAMPLE_FACTORS - d3-Crain_5 Sample source:Galea | Experimental Variable:d3 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-Crain_5.mzXML SUBJECT_SAMPLE_FACTORS - d3-PJI_1 Sample source:Tissue (PJI) | Experimental Variable:d3 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-PJI_1.mzXML SUBJECT_SAMPLE_FACTORS - d3-PJI_2 Sample source:Tissue (PJI) | Experimental Variable:d3 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-PJI_2.mzXML SUBJECT_SAMPLE_FACTORS - d3-PJI_3 Sample source:Tissue (PJI) | Experimental Variable:d3 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-PJI_3.mzXML SUBJECT_SAMPLE_FACTORS - d3-PJI_4 Sample source:Tissue (PJI) | Experimental Variable:d3 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-PJI_4.mzXML SUBJECT_SAMPLE_FACTORS - d3-PJI_5 Sample source:Tissue (PJI) | Experimental Variable:d3 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d3-PJI_5.mzXML SUBJECT_SAMPLE_FACTORS - d7-Crain_1 Sample source:Galea | Experimental Variable:d7 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-Crain_1.mzXML SUBJECT_SAMPLE_FACTORS - d7-Crain_2 Sample source:Galea | Experimental Variable:d7 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-Crain_2.mzXML SUBJECT_SAMPLE_FACTORS - d7-Crain_3 Sample source:Galea | Experimental Variable:d7 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-Crain_3.mzXML SUBJECT_SAMPLE_FACTORS - d7-Crain_4 Sample source:Galea | Experimental Variable:d7 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-Crain_4.mzXML SUBJECT_SAMPLE_FACTORS - d7-Crain_5 Sample source:Galea | Experimental Variable:d7 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-Crain_5.mzXML SUBJECT_SAMPLE_FACTORS - d7-PJI_1 Sample source:Tissue (PJI) | Experimental Variable:d7 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-PJI_1.mzXML SUBJECT_SAMPLE_FACTORS - d7-PJI_2 Sample source:Tissue (PJI) | Experimental Variable:d7 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-PJI_2.mzXML SUBJECT_SAMPLE_FACTORS - d7-PJI_3 Sample source:Tissue (PJI) | Experimental Variable:d7 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-PJI_3.mzXML SUBJECT_SAMPLE_FACTORS - d7-PJI_4 Sample source:Tissue (PJI) | Experimental Variable:d7 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-PJI_4.mzXML SUBJECT_SAMPLE_FACTORS - d7-PJI_5 Sample source:Tissue (PJI) | Experimental Variable:d7 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d7-PJI_5.mzXML SUBJECT_SAMPLE_FACTORS - d14-Crain_1 Sample source:Galea | Experimental Variable:d14 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-Crain_1.mzXML SUBJECT_SAMPLE_FACTORS - d14-Crain_2 Sample source:Galea | Experimental Variable:d14 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-Crain_2.mzXML SUBJECT_SAMPLE_FACTORS - d14-Crain_3 Sample source:Galea | Experimental Variable:d14 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-Crain_3.mzXML SUBJECT_SAMPLE_FACTORS - d14-Crain_4 Sample source:Galea | Experimental Variable:d14 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-Crain_4.mzXML SUBJECT_SAMPLE_FACTORS - d14-Crain_5 Sample source:Galea | Experimental Variable:d14 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-Crain_5.mzXML SUBJECT_SAMPLE_FACTORS - d14-PJI_1 Sample source:Tissue (PJI) | Experimental Variable:d14 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-PJI_1.mzXML SUBJECT_SAMPLE_FACTORS - d14-PJI_2 Sample source:Tissue (PJI) | Experimental Variable:d14 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-PJI_2.mzXML SUBJECT_SAMPLE_FACTORS - d14-PJI_3 Sample source:Tissue (PJI) | Experimental Variable:d14 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-PJI_3.mzXML SUBJECT_SAMPLE_FACTORS - d14-PJI_4 Sample source:Tissue (PJI) | Experimental Variable:d14 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-PJI_4.mzXML SUBJECT_SAMPLE_FACTORS - d14-PJI_5 Sample source:Tissue (PJI) | Experimental Variable:d14 Type=Intracellular; RAW_FILE_NAME(Raw file name)=d14-PJI_5.mzXML #COLLECTION CO:COLLECTION_SUMMARY The intracellular metabolome of granulocytes isolated from the galea and PJI CO:COLLECTION_SUMMARY tissue of mouse craniotomy and PJI models, respectively, were profiled by CO:COLLECTION_SUMMARY LC-HRMS. Fifteen mice were infected for each biofilm model (craniotomy and PJI) CO:COLLECTION_SUMMARY with single-cell suspensions prepared by pooling tissue homogenates from 3 mice CO:COLLECTION_SUMMARY into one sample, resulting in 5 biological replicates. Samples were then CO:COLLECTION_SUMMARY enriched for granulocytes using Ly6G magnetic beads (Miltenyi Biotec, Bergishch CO:COLLECTION_SUMMARY Gladbach, Germany). Magnetic beads were utilized for purification instead of CO:COLLECTION_SUMMARY FACS to limit alterations in cellular metabolism that may occur with extended CO:COLLECTION_SUMMARY processing times. Samples were washed twice in 1X PBS, resuspended in 80% MetOH, CO:COLLECTION_SUMMARY and frozen at -80°C for 15 min. Metabolomics was also performed on supernatants CO:COLLECTION_SUMMARY prepared from brain, galea, and PJI tissue homogenates at days 3, 7, and 14 CO:COLLECTION_SUMMARY post-infection. Similar to granulocyte intracellular metabolite collection, CO:COLLECTION_SUMMARY tissue supernatants were centrifuged at high speed to remove residual cellular CO:COLLECTION_SUMMARY material, combined with 100% MetOH to yield an 80% MetOH solution, and frozen at CO:COLLECTION_SUMMARY -80°C. CO:SAMPLE_TYPE Extracellular infection sites homogenate, intracellular granulocyte #TREATMENT TR:TREATMENT_SUMMARY All mice were group-housed at 21-23°C (22°C average) and 30-70% humidity (55% TR:TREATMENT_SUMMARY average) under 12 h light/dark cycle with free access to food (2019S Teklad TR:TREATMENT_SUMMARY Global 19% Protein Extruded Rodent Diet; Inotiv, West Lafayette, IN) and water. TR:TREATMENT_SUMMARY S. aureus craniotomy infection was established in male and female 8- to TR:TREATMENT_SUMMARY 10-week-old C57BL/6J mice (RRID:IMSR_JAX:000664). Briefly, ketamine and xylazine TR:TREATMENT_SUMMARY were administered to achieve a surgical plane of anesthesia before skin TR:TREATMENT_SUMMARY disinfection with betadine. A midline incision of the scalp was made, whereupon TR:TREATMENT_SUMMARY a bone flap was created using a high-speed pneumatic drill (Stryker Corporation, TR:TREATMENT_SUMMARY Kalamazoo, MI). The bone flap was incubated with 0.5 mL of S. aureus USA300 TR:TREATMENT_SUMMARY LAC13C diluted to 10^6/mL in brain-heart infusion broth at 37°C for 5 min and TR:TREATMENT_SUMMARY subsequently rinsed in PBS, blotted dry on a sterile field, and reimplanted. TR:TREATMENT_SUMMARY This procedure reliably produces an infectious inoculum of 10^3 CFUs per bone TR:TREATMENT_SUMMARY flap, which more accurately models surgical site infection that progresses to TR:TREATMENT_SUMMARY biofilm formation. The scalp incision was closed using 6-0 nylon suture and TR:TREATMENT_SUMMARY animals were continuously monitored until they regained ambulation, and daily TR:TREATMENT_SUMMARY thereafter. S. aureus PJI was established in male and female 8- to 10-week-old TR:TREATMENT_SUMMARY C57BL/6J mice. Mice were anesthetized with ketamine and xylazine prior to skin TR:TREATMENT_SUMMARY disinfection with betadine and initial incision of the quadriceps. A secondary TR:TREATMENT_SUMMARY incision was made to laterally displace the patellar tendon and a burr hole was TR:TREATMENT_SUMMARY created in the distal end of the femur with a 26-gauge needle. An TR:TREATMENT_SUMMARY orthopedic-grade nickel-titanium Kirschner wire (0.6 mm diameter; Custom Wire TR:TREATMENT_SUMMARY Technologies, Port Washington, WI) was inserted through the burr hole into the TR:TREATMENT_SUMMARY medullary canal and 10^3 CFUs of S. aureus USA300 LAC13C were inoculated at the TR:TREATMENT_SUMMARY tip of the wire and the incision was closed with nylon suture. Mice received TR:TREATMENT_SUMMARY buprenorphine SR to alleviate any post-surgical pain and were continuously TR:TREATMENT_SUMMARY monitored until ambulatory and daily thereafter. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY A 13C15N-labeled canonical amino acid (13C15N-CAA) mix (Cambridge Isotope SP:SAMPLEPREP_SUMMARY Laboratories, Andover, MA) was added to the extraction buffer comprised of 80% SP:SAMPLEPREP_SUMMARY MeOH as an internal standard. Samples were then pelleted, and supernatants were SP:SAMPLEPREP_SUMMARY dried by vacuum centrifuge for storage at -80°C. Cell pellets were stored for SP:SAMPLEPREP_SUMMARY protein quantification to normalize metabolite peak areas. Prior to LC-HRMS, SP:SAMPLEPREP_SUMMARY samples were reconstituted in 100 µL of 50% MetOH, pelleted, and the resulting SP:SAMPLEPREP_SUMMARY supernatant was analyzed. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Metabolite separation was performed by liquid chromatography using a XBridge CH:CHROMATOGRAPHY_SUMMARY Amide analytical column (150 × 2.1mm ID; 1.7µm particle size; Waters CH:CHROMATOGRAPHY_SUMMARY Corporation, Milford, MA) and a binary solvent system infused at a flow rate of CH:CHROMATOGRAPHY_SUMMARY 0.3 ml/min. A guard XBridge Amide column (20 × 2.1mm ID; 3.5µm particle CH:CHROMATOGRAPHY_SUMMARY size; Waters Corporation) was connected in front of the analytical column. CH:CHROMATOGRAPHY_SUMMARY Mobile phase A was composed of ammonium acetate and ammonium hydroxide (10 mM CH:CHROMATOGRAPHY_SUMMARY each) containing 5% acetonitrile in LC-MS grade water; mobile phase B was 100% CH:CHROMATOGRAPHY_SUMMARY LC-MS grade acetonitrile. The pH of mobile phase A was adjusted to 8.0 using CH:CHROMATOGRAPHY_SUMMARY glacial acetic acid. The UHPLC pumps were operated in gradient mode. The amide CH:CHROMATOGRAPHY_SUMMARY column was maintained at 40°C, and the autosampler temperature was held at 5°C CH:CHROMATOGRAPHY_SUMMARY throughout data acquisition. The injection volume for all samples was 5 µl with CH:CHROMATOGRAPHY_SUMMARY the 13C15N-CAA mix as the internal standard. CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME Waters ACQUITY UPLC BEH Amide (150 x 2.1mm,1.7um) CH:SOLVENT_A ammonium acetate and ammonium hydroxide (10 mM each) containing 5% acetonitrile CH:SOLVENT_A in LC-MS grade water CH:SOLVENT_B acetonitrile CH:FLOW_GRADIENT B% at initial 85% CH:FLOW_RATE 0.3ml/min CH:COLUMN_TEMPERATURE 40 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Orbitrap Exploris 480 MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS A HRMS Orbitrap (Exploris 480; Thermo Fisher Scientific) was operated in MS:MS_COMMENTS polarity switching mode and was used for untargeted metabolomics in a MS:MS_COMMENTS data-dependent MS/MS acquisition mode (DDA). Electrospray ionization (ESI) MS:MS_COMMENTS parameters were optimized as follows: electrospray ion voltage of -2700V and MS:MS_COMMENTS 3500V in negative and positive mode respectively, ion transfer tube temperature MS:MS_COMMENTS was maintained at 400°C, and m/z scan range was 70-1050 Da. Orbitrap resolution MS:MS_COMMENTS for precursor ion scans were maintained at 240,000 for intracellular and 120,000 MS:MS_COMMENTS for extracellular whereas fragment ion scans were maintained at 120,000 and MS:MS_COMMENTS 60,000, for intracellular and extracellular respectively. Normalized collision MS:MS_COMMENTS energies at 30, 50, and 150% were used for fragmentation and data was acquired MS:MS_COMMENTS in profile mode. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Relative intensities MS_METABOLITE_DATA_START Samples d3-Crain_1 d3-Crain_2 d3-Crain_3 d3-Crain_4 d3-Crain_5 d3-PJI_1 d3-PJI_2 d3-PJI_3 d3-PJI_4 d3-PJI_5 d7-Crain_1 d7-Crain_2 d7-Crain_3 d7-Crain_4 d7-Crain_5 d7-PJI_1 d7-PJI_2 d7-PJI_3 d7-PJI_4 d7-PJI_5 d14-Crain_1 d14-Crain_2 d14-Crain_3 d14-Crain_4 d14-Crain_5 d14-PJI_1 d14-PJI_2 d14-PJI_3 d14-PJI_4 d14-PJI_5 Factors Sample source:Galea | Experimental Variable:d3 Sample source:Galea | Experimental Variable:d3 Sample source:Galea | Experimental Variable:d3 Sample source:Galea | Experimental Variable:d3 Sample source:Galea | Experimental Variable:d3 Sample source:Tissue (PJI) | Experimental Variable:d3 Sample source:Tissue (PJI) | Experimental Variable:d3 Sample source:Tissue (PJI) | Experimental Variable:d3 Sample source:Tissue (PJI) | Experimental Variable:d3 Sample source:Tissue (PJI) | Experimental Variable:d3 Sample source:Galea | Experimental Variable:d7 Sample source:Galea | Experimental Variable:d7 Sample source:Galea | Experimental Variable:d7 Sample source:Galea | Experimental Variable:d7 Sample source:Galea | Experimental Variable:d7 Sample source:Tissue (PJI) | Experimental Variable:d7 Sample source:Tissue (PJI) | Experimental Variable:d7 Sample source:Tissue (PJI) | Experimental Variable:d7 Sample source:Tissue (PJI) | Experimental Variable:d7 Sample source:Tissue (PJI) | Experimental Variable:d7 Sample source:Galea | Experimental Variable:d14 Sample source:Galea | Experimental Variable:d14 Sample source:Galea | Experimental Variable:d14 Sample source:Galea | Experimental Variable:d14 Sample source:Galea | Experimental Variable:d14 Sample source:Tissue (PJI) | Experimental Variable:d14 Sample source:Tissue (PJI) | Experimental Variable:d14 Sample source:Tissue (PJI) | Experimental Variable:d14 Sample source:Tissue (PJI) | Experimental Variable:d14 Sample source:Tissue (PJI) | Experimental Variable:d14 (1S,2R,5S)-2-Isopropyl-5-methylcyclohexyl 3-oxobutanoate 3.07E+04 3.39E+04 2.03E+04 1.14E+04 1.89E+04 3.31E+04 3.52E+04 3.36E+04 3.01E+04 1.56E+04 9.88E+03 3.40E+04 9.33E+03 2.29E+04 3.48E+04 2.28E+04 3.00E+04 3.17E+04 2.48E+04 7.21E+04 1.63E+05 3.60E+04 3.13E+04 8.62E+04 8.50E+04 1.68E+04 1.40E+04 3.69E+04 1.96E+04 8.10E+03 (2E)-decenoic acid 2.17E+04 2.05E+04 1.63E+04 7.97E+03 1.80E+04 2.35E+04 2.09E+04 2.31E+04 1.76E+04 1.60E+04 8.43E+03 2.45E+04 9.32E+03 1.73E+04 1.95E+04 1.19E+04 1.25E+04 2.63E+04 2.13E+04 5.44E+04 6.67E+04 1.77E+04 1.45E+04 6.24E+03 6.97E+04 1.53E+04 6.74E+03 2.73E+04 1.73E+04 4.45E+03 Dihydroartemisinin 4.74E+04 5.41E+04 5.49E+04 2.87E+04 2.82E+04 8.73E+04 5.55E+04 9.83E+04 6.08E+04 3.07E+04 2.07E+04 6.12E+04 1.96E+04 3.81E+04 6.31E+04 4.39E+04 6.05E+04 6.23E+04 6.41E+04 1.85E+05 2.21E+05 4.81E+04 9.73E+04 9.67E+04 1.51E+05 4.61E+04 2.26E+04 7.86E+04 5.73E+04 1.74E+04 (5E,9Z)-farnesyl acetone 1.78E+04 1.27E+04 1.13E+04 6.99E+03 8.38E+03 2.33E+04 1.63E+04 1.69E+04 1.63E+04 1.08E+04 4.52E+03 1.13E+04 4.17E+03 1.13E+04 1.29E+04 1.21E+04 9.01E+03 1.41E+04 8.08E+03 2.42E+04 7.10E+04 8.64E+03 1.78E+04 4.07E+04 4.88E+04 1.27E+04 9.86E+03 2.97E+04 1.33E+04 4.88E+03 (R)-3-hydroxybutyrylcarnitine 5.72E+04 4.72E+04 3.23E+04 1.63E+04 5.40E+04 5.26E+04 4.93E+04 5.03E+04 6.06E+04 6.05E+04 2.98E+04 3.07E+04 4.68E+04 2.44E+04 2.88E+04 3.69E+04 2.50E+04 3.36E+04 4.46E+04 3.85E+04 1.90E+04 1.49E+04 2.48E+04 7.13E+02 7.51E+03 7.33E+04 1.76E+04 2.34E+04 7.83E+04 1.24E+04 15,16-DiHODE 4.48E+04 3.51E+04 2.57E+04 1.34E+04 2.92E+04 9.36E+04 4.25E+04 5.09E+04 5.21E+04 3.07E+04 1.62E+04 5.51E+04 1.15E+04 3.76E+04 4.31E+04 2.94E+04 4.93E+04 4.54E+04 4.25E+04 9.70E+04 2.85E+05 4.96E+04 6.22E+03 1.21E+05 1.01E+05 3.96E+04 2.32E+04 5.57E+04 5.14E+04 1.38E+04 1-Linoleoyl-2-Hydroxy-sn-glycero-3-PC 4.27E+04 3.62E+04 6.27E+04 5.60E+04 6.96E+04 3.05E+04 1.54E+04 2.32E+04 2.37E+04 1.28E+04 1.89E+04 3.62E+04 1.32E+04 2.82E+04 2.70E+04 2.52E+04 2.28E+04 1.16E+04 1.56E+04 2.51E+04 1.31E+05 3.64E+04 3.98E+04 2.11E+05 6.10E+04 5.24E+04 1.02E+04 1.90E+04 3.52E+04 1.13E+04 1-Methylhistamine 1.62E+05 1.64E+05 4.57E+03 4.70E+04 4.69E+03 2.93E+05 2.52E+05 3.12E+05 3.15E+05 2.97E+05 1.31E+05 1.53E+05 9.29E+04 1.90E+05 1.33E+05 1.71E+05 2.03E+05 1.52E+05 2.31E+05 2.38E+05 1.40E+05 1.12E+05 1.47E+05 4.20E+03 9.27E+04 2.18E+05 1.22E+05 3.01E+05 3.55E+05 8.48E+04 1-Methyl-N-(9-methyl-9-azabicyclo[3.3.1]non-3-yl)-1H-indazole-3-carboxamide 3.92E+04 4.19E+04 2.90E+04 1.48E+04 2.51E+04 7.12E+04 4.37E+04 4.70E+04 5.35E+04 2.53E+04 1.67E+04 4.39E+04 1.40E+04 6.28E+04 4.43E+04 3.51E+04 5.22E+04 5.79E+04 4.66E+04 1.23E+05 1.87E+05 3.39E+04 4.79E+04 7.83E+04 1.13E+05 3.98E+04 2.82E+04 6.70E+04 4.61E+04 1.24E+04 1-Stearoyl-2-hydroxy-sn-glycero-3-PE 1.10E+04 9.25E+03 7.80E+03 6.20E+03 8.96E+03 6.31E+03 3.60E+03 4.41E+03 5.98E+03 3.97E+03 4.75E+03 7.33E+03 3.63E+03 1.37E+04 6.10E+03 4.30E+03 4.38E+03 5.09E+03 3.39E+03 3.70E+03 1.78E+04 6.53E+03 7.65E+03 1.70E+04 8.00E+03 9.72E+03 2.45E+03 7.40E+03 4.66E+03 2.14E+03 2-(Diethylamino)ethanol 3.65E+05 6.54E+05 9.36E+05 9.77E+05 1.78E+06 6.20E+06 1.39E+06 1.16E+05 6.02E+06 5.02E+04 2.42E+04 5.75E+04 8.28E+03 9.34E+04 3.16E+06 3.04E+04 5.49E+04 6.68E+04 4.60E+04 1.53E+05 3.13E+05 3.80E+06 1.69E+06 5.26E+06 5.82E+06 5.15E+04 3.08E+04 1.43E+05 6.71E+05 8.26E+03 2,4-dihydroxyheptadec-16-ynyl acetate 3.23E+04 3.06E+04 2.16E+04 1.12E+04 2.41E+04 4.49E+04 2.78E+04 4.16E+04 4.64E+04 2.45E+04 1.13E+04 3.17E+04 1.25E+04 3.25E+04 3.75E+04 2.56E+04 2.71E+04 4.28E+04 4.31E+04 1.15E+05 1.34E+05 3.08E+04 3.90E+04 4.98E+04 1.02E+05 3.29E+04 2.11E+04 4.72E+04 3.47E+04 1.24E+04 2-[4-(3-Hydroxypropyl)-2-methoxyphenoxy]-1,3-propanediol 1.58E+04 2.93E+04 2.24E+04 1.39E+04 1.62E+04 4.24E+04 2.72E+04 3.87E+04 3.50E+04 1.95E+04 9.45E+03 2.65E+04 8.44E+03 2.08E+04 2.64E+04 2.73E+04 5.05E+04 4.01E+04 2.65E+04 7.28E+04 1.18E+05 3.45E+04 3.78E+04 5.98E+04 6.20E+04 2.08E+04 1.45E+04 4.47E+04 2.17E+04 1.22E+04 2'-Deoxycytidine 7.56E+03 1.95E+02 6.26E+03 4.19E+03 2.88E+03 3.96E+03 2.84E+03 1.03E+04 6.98E+03 1.64E+04 1.26E+04 3.15E+04 1.26E+04 4.46E+02 3.36E+03 2.61E+04 4.53E+04 2.66E+04 3.91E+04 2.00E+04 1.98E+04 4.20E+03 6.33E+03 8.65E+02 1.84E+03 7.34E+04 3.08E+04 2.96E+04 1.30E+04 9.21E+03 2-Methoxypyrazine 1.81E+05 1.48E+05 7.07E+04 4.92E+04 5.13E+04 1.95E+05 1.06E+05 1.52E+05 1.75E+05 1.37E+05 3.67E+04 9.61E+04 2.70E+04 2.73E+05 5.46E+04 9.30E+04 1.28E+05 1.09E+05 1.10E+05 9.25E+04 1.63E+05 6.18E+04 6.20E+04 8.72E+03 1.32E+05 8.86E+04 3.24E+04 2.04E+05 1.14E+05 2.22E+04 2-Octenoic acid 3.97E+03 1.10E+04 7.68E+03 6.16E+03 5.68E+03 1.49E+04 9.86E+03 1.13E+04 1.46E+04 5.62E+03 4.38E+03 1.82E+04 2.90E+03 1.60E+04 1.15E+04 1.16E+04 1.45E+04 1.03E+04 1.34E+04 3.62E+04 5.96E+04 9.64E+03 1.28E+04 1.53E+04 4.80E+04 1.15E+04 3.89E+03 2.19E+04 1.66E+04 3.58E+03 3,4-Dihydroxyphenylglycol 5.09E+03 4.75E+03 3.64E+03 2.03E+03 2.28E+03 7.11E+03 4.34E+03 4.57E+03 4.82E+03 2.47E+03 2.19E+03 6.19E+03 1.96E+03 2.26E+03 4.10E+03 2.78E+03 3.86E+03 4.75E+03 4.73E+03 1.28E+04 2.02E+04 4.89E+03 2.72E+03 4.33E+03 6.02E+03 4.23E+03 2.65E+03 4.55E+03 6.00E+03 2.00E+03 3,4-Dimethyl-5-pentyl-2-furanpropanoic acid 1.10E+05 8.55E+04 6.64E+04 3.39E+04 6.44E+04 1.49E+05 8.05E+04 1.12E+05 8.67E+04 5.99E+04 3.35E+04 1.28E+05 2.59E+04 7.02E+04 7.19E+04 5.97E+04 8.86E+04 9.07E+04 8.12E+04 2.07E+05 3.75E+05 8.42E+04 9.51E+04 1.79E+05 2.23E+05 6.58E+04 5.18E+04 1.15E+05 9.87E+04 3.29E+04 3-[(3-Methylbutyl)nitrosoamino]-2-butanone 4.57E+03 5.85E+03 7.60E+03 4.55E+03 6.81E+03 2.71E+04 5.98E+03 5.30E+03 1.24E+04 3.96E+03 8.93E+03 1.63E+04 3.13E+03 3.26E+04 1.26E+04 1.79E+04 1.17E+04 1.63E+04 2.52E+04 5.31E+04 2.01E+05 3.02E+04 1.56E+04 8.41E+04 1.19E+05 1.22E+04 9.56E+03 4.67E+04 1.81E+04 5.10E+03 4-[(2-Isopropyl-5-methylcyclohexyl)oxy]-4-oxobutanoic acid 6.26E+04 3.79E+04 2.42E+04 1.64E+04 2.29E+04 4.26E+04 3.38E+04 4.85E+04 4.32E+04 2.71E+04 1.24E+04 3.64E+04 1.11E+04 3.18E+04 4.45E+04 2.47E+04 4.22E+04 3.78E+04 3.46E+04 1.03E+05 1.01E+05 2.62E+04 3.38E+04 6.39E+04 1.16E+05 2.89E+04 1.68E+04 4.29E+04 3.93E+04 1.29E+04 4-Aminobenzoic acid 7.66E+03 9.22E+03 5.54E+03 9.05E+03 8.18E+03 5.90E+03 4.82E+03 3.95E+03 5.39E+03 2.82E+03 3.69E+03 3.86E+03 4.95E+03 5.13E+03 4.35E+03 3.97E+03 4.72E+03 3.36E+03 4.83E+03 5.26E+03 1.75E+03 2.70E+03 4.85E+03 2.80E+03 5.42E+03 7.82E+03 2.23E+03 6.59E+03 1.24E+04 1.89E+03 4-Hydroxy-5-methoxy-4-[2-methyl-3-(3-methyl-2-buten-1-yl)-2-oxiranyl]-1-oxaspiro[2.5]octan-6-one 4.89E+04 4.19E+04 3.03E+04 1.40E+04 2.61E+04 9.92E+04 4.98E+04 5.43E+04 5.48E+04 2.53E+04 1.25E+04 5.82E+04 1.57E+04 4.40E+04 5.10E+04 4.71E+04 4.33E+04 5.72E+04 3.47E+04 1.40E+05 1.99E+05 4.38E+04 4.72E+04 1.00E+05 1.47E+05 4.31E+04 2.36E+04 6.44E+04 3.88E+04 1.79E+04 5,5'-[Disulfanediylbis({3-[(carboxymethyl)amino]-3-oxo-1,2-propanediyl}imino)]bis(2-amino-5-oxopentanoic acid) 1.13E+04 1.37E+04 1.98E+04 2.62E+04 1.93E+04 3.57E+04 3.76E+04 4.76E+04 5.25E+04 7.67E+04 2.42E+04 2.15E+04 5.30E+04 3.86E+04 2.21E+04 4.09E+04 4.91E+04 4.10E+04 4.70E+04 4.27E+04 1.97E+04 1.47E+04 2.52E+04 1.41E+03 9.38E+03 1.09E+05 2.25E+04 5.53E+04 9.41E+04 1.72E+04 5-Isobutyl-2-oxotetrahydro-3-furancarboxylic acid 1.07E+04 1.18E+04 8.20E+03 3.88E+03 6.51E+03 1.91E+04 1.13E+04 1.64E+04 1.08E+04 2.44E+03 1.98E+03 1.85E+04 1.80E+03 1.09E+04 9.82E+03 1.52E+04 1.52E+04 1.09E+04 1.45E+04 2.53E+04 1.22E+05 1.20E+04 1.88E+04 2.60E+04 4.54E+04 1.10E+04 5.31E+03 1.96E+04 1.82E+04 4.33E+03 6-Methoxyelemicin 8.74E+04 6.11E+04 2.99E+04 2.97E+04 5.63E+04 1.04E+05 1.26E+05 9.39E+04 7.63E+04 3.04E+04 2.56E+04 9.99E+04 2.13E+04 5.80E+04 6.70E+04 5.88E+04 8.49E+04 8.58E+04 8.38E+04 2.10E+05 2.56E+05 6.66E+04 9.91E+04 1.22E+05 2.40E+05 5.15E+04 3.03E+04 8.92E+04 6.28E+04 2.61E+04 7-Hydroxymyoporone 1.45E+05 7.23E+04 6.74E+04 4.03E+04 4.96E+04 1.23E+05 7.96E+04 1.04E+05 1.18E+05 5.61E+04 2.39E+04 8.83E+04 3.27E+04 6.68E+04 7.69E+04 1.01E+05 8.99E+04 1.03E+05 1.07E+05 2.53E+05 3.87E+05 7.31E+04 1.15E+05 1.67E+05 1.77E+05 8.58E+04 4.23E+04 1.31E+05 8.63E+04 3.00E+04 Acetylcarnitine 2.59E+06 2.74E+06 1.19E+06 1.02E+06 5.78E+05 4.23E+06 3.58E+06 4.36E+06 3.76E+06 4.26E+06 1.79E+06 2.41E+06 2.16E+06 2.09E+06 1.94E+06 3.18E+06 3.48E+06 3.17E+06 3.40E+06 3.83E+06 1.84E+06 1.07E+06 1.81E+06 2.50E+04 1.33E+06 5.36E+06 1.63E+06 3.40E+06 6.90E+06 1.25E+06 Adenosine monophosphate 3.32E+04 2.66E+04 2.24E+04 2.59E+04 1.94E+04 2.44E+04 2.11E+04 2.37E+04 2.54E+04 3.80E+04 2.19E+04 2.41E+04 3.32E+04 3.37E+04 1.88E+04 2.32E+04 2.24E+04 2.83E+04 2.82E+04 2.53E+04 2.71E+04 1.13E+04 2.15E+04 7.88E+03 2.15E+04 5.17E+04 1.46E+04 2.99E+04 4.81E+04 7.41E+03 Alpha-GPC 2.90E+04 2.95E+04 1.67E+04 2.89E+04 1.99E+04 1.60E+04 1.20E+04 1.79E+04 1.42E+04 1.28E+04 1.78E+04 2.52E+04 1.85E+04 1.97E+04 2.18E+04 2.38E+04 1.97E+04 4.43E+04 2.60E+04 1.91E+04 6.37E+04 3.02E+04 3.20E+04 1.49E+04 3.37E+04 3.03E+04 8.56E+03 1.96E+04 3.11E+04 6.73E+03 Aminolevulinic acid 1.37E+03 2.87E+03 4.13E+03 3.28E+03 5.02E+03 2.14E+03 1.52E+03 1.48E+03 2.22E+03 1.59E+03 5.44E+02 4.89E+02 1.20E+03 7.74E+02 1.11E+03 1.08E+03 6.86E+02 5.22E+02 1.19E+03 1.06E+03 1.23E+03 1.10E+03 7.55E+02 2.63E+03 5.74E+02 2.39E+03 4.37E+02 9.55E+02 2.60E+03 4.76E+02 Butenylcarnitine 2.63E+04 3.06E+04 2.93E+04 7.58E+03 1.35E+04 7.23E+04 3.88E+04 4.46E+04 4.36E+04 2.09E+04 8.54E+03 4.54E+04 9.43E+03 1.81E+04 2.24E+04 1.90E+04 7.70E+04 2.82E+04 3.75E+04 5.92E+04 2.72E+05 3.78E+04 2.54E+04 1.08E+05 4.45E+04 2.18E+04 8.33E+03 2.78E+04 3.08E+04 2.13E+04 Butylparaben 6.79E+04 4.57E+04 2.51E+04 2.05E+04 3.14E+04 4.91E+04 3.71E+04 5.16E+04 4.91E+04 2.53E+04 1.70E+04 6.11E+04 1.40E+04 4.13E+04 4.83E+04 4.31E+04 4.32E+04 6.15E+04 4.69E+04 1.28E+05 2.79E+05 4.53E+04 4.81E+04 9.57E+04 1.27E+05 3.20E+04 2.03E+04 7.42E+04 4.76E+04 1.71E+04 C14-Carnitine 2.01E+04 2.59E+04 1.87E+04 9.91E+03 1.18E+04 3.02E+05 7.06E+04 4.67E+04 9.30E+04 7.48E+04 7.51E+03 2.87E+04 1.66E+04 3.18E+04 1.28E+04 2.34E+04 3.48E+04 2.82E+04 4.53E+04 1.16E+05 1.67E+04 1.01E+04 2.73E+04 5.18E+03 4.20E+04 7.26E+04 2.22E+04 7.07E+04 7.59E+04 5.76E+03 C4-Carnitine 5.10E+04 5.85E+04 3.02E+04 3.72E+04 3.96E+04 6.31E+04 6.16E+04 5.60E+04 7.50E+04 6.86E+04 7.76E+03 1.23E+04 2.63E+04 2.72E+04 2.29E+04 2.42E+04 4.54E+04 7.34E+04 1.71E+04 4.19E+04 3.20E+04 1.50E+04 2.01E+04 5.33E+03 2.96E+04 6.13E+04 1.75E+04 1.68E+04 8.09E+04 1.50E+04 cis-3-Hexenyl Pyruvate 5.97E+04 4.69E+04 2.74E+04 7.45E+03 3.95E+04 4.70E+04 3.11E+04 4.86E+04 4.41E+04 2.34E+04 2.42E+04 7.45E+04 1.52E+04 4.78E+04 5.39E+04 3.72E+04 8.89E+04 5.84E+04 4.48E+04 1.03E+05 2.40E+05 3.55E+04 5.17E+04 9.66E+04 1.24E+05 2.41E+04 2.04E+04 6.53E+04 3.35E+04 1.69E+04 Colfosceril palmitate 2.68E+05 4.33E+05 1.07E+05 1.20E+04 7.27E+04 6.57E+04 5.96E+04 1.05E+05 1.20E+05 7.30E+04 5.03E+04 2.11E+04 2.83E+04 1.13E+04 1.13E+05 1.07E+05 5.08E+04 1.59E+05 1.04E+05 1.80E+05 8.62E+04 2.91E+04 2.69E+04 1.18E+03 1.58E+04 1.02E+05 5.07E+04 1.44E+04 2.57E+04 4.81E+03 Cyclohexylamine 2.52E+05 1.02E+05 1.53E+05 1.57E+05 2.65E+05 1.02E+06 6.50E+05 4.47E+05 7.11E+05 6.19E+05 5.73E+04 2.05E+05 1.04E+05 6.34E+05 5.11E+05 2.74E+05 2.28E+04 2.12E+04 2.20E+05 4.89E+04 4.11E+06 4.82E+05 2.47E+05 8.69E+05 3.76E+05 2.32E+04 1.19E+04 4.78E+05 2.39E+05 8.70E+04 Cytosine 1.11E+04 7.76E+03 5.63E+03 2.18E+03 2.71E+03 5.73E+03 4.86E+03 1.04E+04 6.54E+03 1.47E+04 1.32E+04 3.28E+04 1.22E+04 5.69E+04 4.20E+03 2.62E+04 4.64E+04 3.23E+04 3.74E+04 2.30E+04 2.54E+04 5.54E+03 9.96E+02 8.03E+02 6.37E+03 6.88E+04 2.94E+04 3.47E+04 1.21E+04 8.38E+03 Deanol 2.84E+05 1.72E+05 1.20E+04 1.17E+04 1.64E+04 1.31E+06 3.81E+05 3.43E+05 3.11E+05 2.25E+05 1.54E+05 5.50E+05 7.10E+04 3.60E+05 2.81E+05 4.15E+05 2.63E+05 2.55E+05 4.44E+05 5.95E+05 2.80E+06 4.48E+05 2.32E+05 4.59E+04 2.28E+05 2.49E+05 1.87E+05 4.14E+05 2.44E+05 1.07E+05 Diethylamine 9.98E+04 8.62E+04 6.72E+04 5.32E+04 7.98E+04 2.23E+05 1.20E+05 1.27E+05 1.47E+05 8.07E+04 1.71E+04 3.96E+04 2.77E+04 1.13E+05 1.06E+05 2.34E+04 5.22E+04 6.28E+04 3.45E+04 1.51E+05 3.54E+05 7.74E+04 1.17E+05 3.09E+05 3.83E+05 3.77E+04 1.89E+04 1.54E+05 8.87E+04 1.27E+04 DL-Citrulline 2.40E+04 1.89E+04 2.13E+04 2.14E+04 1.33E+04 2.54E+04 1.53E+04 1.99E+04 2.21E+04 2.34E+04 1.35E+04 1.13E+04 1.25E+04 1.28E+04 8.44E+03 1.99E+04 1.64E+04 1.04E+04 1.26E+04 9.08E+03 1.79E+04 9.66E+03 1.20E+04 2.38E+03 9.84E+03 2.84E+04 1.06E+04 1.93E+04 3.25E+04 4.74E+03 DL-Glutamic acid 1.48E+05 1.39E+05 1.14E+05 1.39E+05 7.87E+04 1.07E+05 8.61E+04 1.02E+05 1.23E+05 1.26E+05 7.51E+04 8.87E+04 7.53E+04 7.18E+04 5.42E+04 6.68E+04 5.43E+04 7.87E+04 5.62E+04 5.50E+04 9.66E+04 3.67E+04 6.76E+04 1.22E+04 4.50E+04 1.55E+05 2.63E+04 4.52E+04 1.07E+05 1.41E+04 D-PANTOTHENIC ACID 5.32E+03 8.42E+03 1.07E+04 1.27E+04 9.04E+03 3.05E+03 2.12E+03 1.30E+03 3.79E+03 1.70E+03 1.16E+03 1.77E+03 2.86E+03 2.85E+03 3.79E+03 1.45E+03 6.37E+02 3.25E+02 1.48E+03 1.17E+03 1.03E+04 2.81E+03 3.69E+03 1.09E+03 5.50E+03 8.21E+03 1.71E+03 4.06E+03 5.14E+03 5.36E+02 Ethylenediaminetetraacetic acid 2.44E+05 1.56E+05 5.85E+05 3.35E+05 5.53E+05 1.84E+05 9.65E+04 1.12E+05 9.01E+04 4.87E+04 3.12E+04 4.06E+04 2.32E+04 1.47E+04 5.86E+04 2.05E+04 1.52E+04 6.31E+03 1.41E+04 2.14E+04 2.64E+05 3.33E+04 6.02E+04 5.55E+05 4.75E+05 1.69E+04 3.93E+03 3.41E+03 2.07E+04 5.49E+03 g-Butyrobetaine 6.14E+04 3.78E+04 2.50E+04 4.44E+04 2.96E+04 5.43E+04 3.28E+04 3.57E+04 4.79E+04 4.91E+04 2.44E+04 2.37E+04 2.84E+04 3.45E+04 1.65E+04 3.05E+04 3.88E+04 2.88E+04 3.79E+04 4.31E+04 2.63E+04 1.75E+04 2.20E+04 2.82E+03 1.46E+04 6.47E+04 1.82E+04 3.50E+04 6.60E+04 1.21E+04 HEPES 3.50E+05 3.81E+06 1.22E+06 1.59E+04 3.40E+05 3.25E+03 1.47E+04 1.16E+04 2.25E+03 1.58E+02 9.30E+04 4.32E+05 1.26E+05 3.41E+05 4.02E+05 1.74E+05 2.89E+04 6.21E+04 5.26E+03 1.38E+03 1.15E+06 3.45E+05 5.26E+05 3.74E+02 1.46E+06 2.00E+04 8.00E+02 1.94E+04 4.09E+04 1.38E+04 hercynine 1.40E+04 1.59E+04 9.62E+03 5.29E+03 4.82E+03 1.63E+04 1.16E+04 1.64E+04 1.43E+04 1.28E+04 1.20E+04 1.28E+04 1.03E+04 1.70E+04 1.02E+04 1.69E+04 1.97E+04 1.47E+04 1.82E+04 2.36E+04 1.22E+04 6.42E+03 1.06E+04 1.71E+03 9.07E+03 1.80E+04 9.76E+03 3.44E+04 2.33E+04 5.43E+03 Hexylresorcinol 1.20E+05 1.00E+05 9.16E+04 3.80E+04 6.94E+04 1.40E+05 1.20E+05 1.42E+05 1.22E+05 6.96E+04 4.44E+04 1.39E+05 3.31E+04 1.16E+05 1.01E+05 7.79E+04 1.17E+05 1.47E+05 1.39E+05 2.60E+05 5.56E+05 7.68E+04 1.29E+05 2.01E+05 2.73E+05 1.03E+05 6.23E+04 1.50E+05 1.00E+05 3.29E+04 His-pro 1.60E+03 1.81E+03 1.80E+03 1.82E+03 1.41E+03 5.97E+03 1.46E+03 1.12E+03 2.55E+03 2.80E+02 2.21E+02 7.14E+02 3.02E+02 3.38E+02 1.42E+03 1.01E+03 7.63E+02 4.66E+02 5.29E+02 2.40E+03 3.72E+03 1.10E+03 1.21E+03 1.13E+03 7.59E+03 2.04E+03 4.98E+02 6.59E+02 1.65E+03 3.36E+02 Histamine 1.06E+06 9.02E+05 3.27E+05 2.52E+05 8.57E+04 6.53E+05 5.80E+05 7.13E+05 8.21E+05 8.54E+05 8.38E+05 6.97E+05 4.06E+05 1.21E+06 3.64E+05 5.82E+05 7.49E+05 6.42E+05 4.03E+05 3.10E+05 2.52E+05 1.80E+05 2.29E+05 7.98E+03 7.35E+04 9.50E+05 2.50E+05 9.84E+05 6.12E+05 1.05E+05 Inosinic acid 3.40E+03 2.79E+03 2.63E+04 2.27E+04 3.23E+04 2.15E+04 2.16E+04 2.54E+04 2.85E+04 1.38E+04 1.41E+04 5.53E+04 1.79E+04 5.48E+04 6.71E+04 3.31E+04 3.16E+04 1.04E+04 2.24E+04 3.98E+04 2.31E+05 6.26E+04 8.78E+04 1.43E+05 4.11E+05 1.92E+04 8.19E+03 1.75E+04 2.51E+04 7.02E+03 Inspra 4.60E+05 3.09E+05 1.99E+05 9.08E+04 2.33E+05 7.74E+05 1.39E+05 4.39E+05 6.31E+05 3.13E+05 3.55E+05 3.06E+05 2.17E+05 1.10E+06 4.51E+05 2.86E+05 3.43E+05 7.95E+05 7.47E+05 1.82E+06 1.68E+06 2.89E+05 4.02E+05 3.78E+05 6.41E+05 2.11E+05 2.25E+05 5.17E+05 1.41E+06 7.00E+04 L-(-)-Serine 2.83E+04 3.22E+04 2.60E+04 2.36E+04 1.63E+04 4.75E+04 2.64E+04 2.86E+04 4.20E+04 3.65E+04 2.47E+04 2.38E+04 2.81E+04 2.04E+04 2.00E+04 2.64E+04 2.60E+04 1.65E+04 2.55E+04 1.64E+04 2.62E+04 1.45E+04 2.14E+04 5.42E+03 1.79E+04 4.86E+04 1.44E+04 2.36E+04 5.99E+04 9.57E+03 L-(+)-Arginine 6.02E+04 1.60E+05 1.21E+05 1.28E+05 1.11E+05 7.16E+04 4.62E+04 5.04E+04 6.15E+04 6.05E+04 4.72E+04 5.60E+04 5.29E+04 3.98E+04 4.55E+04 4.80E+04 3.69E+04 3.05E+04 3.85E+04 3.60E+04 1.23E+05 4.97E+04 7.12E+04 5.96E+04 1.04E+05 8.46E+04 2.56E+04 5.36E+04 8.83E+04 1.67E+04 L-(+)-Aspartic acid 4.91E+04 4.74E+04 2.70E+04 3.01E+04 1.70E+04 3.78E+04 3.05E+04 3.69E+04 3.31E+04 3.84E+04 2.09E+04 2.62E+04 2.22E+04 2.06E+04 1.45E+04 2.75E+04 2.21E+04 3.67E+04 2.14E+04 1.95E+04 3.32E+04 1.52E+04 2.54E+04 4.90E+03 1.66E+04 5.20E+04 1.16E+04 2.08E+04 4.20E+04 7.12E+03 L-(+)-ERGOTHIONEINE 5.23E+04 7.77E+04 2.64E+03 6.74E+02 5.28E+02 1.64E+05 5.56E+04 1.64E+05 1.85E+05 1.80E+05 1.01E+05 1.09E+05 1.92E+05 1.89E+04 1.06E+05 1.97E+05 7.56E+04 4.50E+04 1.43E+05 1.48E+05 7.41E+04 8.16E+04 1.50E+05 6.81E+02 5.29E+04 4.64E+05 1.12E+05 1.03E+05 4.91E+05 9.59E+04 L-(+)-glutamine 5.02E+04 1.07E+05 5.94E+04 5.91E+04 3.11E+04 3.91E+04 3.42E+04 3.94E+04 5.40E+04 5.81E+04 4.27E+04 4.25E+04 4.19E+04 4.32E+04 3.43E+04 4.16E+04 3.43E+04 3.05E+04 2.86E+04 1.58E+04 8.35E+04 2.43E+04 3.51E+04 2.50E+04 4.46E+04 7.05E+04 1.88E+04 3.99E+04 5.76E+04 1.30E+04 L-(+)-Leucine 2.17E+05 4.21E+05 2.43E+05 3.64E+05 1.89E+05 2.63E+05 1.84E+05 2.22E+05 3.50E+05 3.41E+05 2.10E+05 2.00E+05 2.29E+05 2.20E+05 1.94E+05 2.31E+05 2.10E+05 1.85E+05 2.13E+05 1.62E+05 3.16E+05 1.58E+05 1.73E+05 1.08E+05 2.09E+05 4.28E+05 1.23E+05 1.44E+05 4.94E+05 8.26E+04 L-(+)-Valine 1.54E+06 2.90E+06 1.95E+06 2.07E+06 1.86E+06 2.93E+06 2.00E+06 2.67E+06 2.65E+06 2.08E+06 1.52E+06 3.33E+06 1.24E+06 2.05E+06 2.01E+06 2.19E+06 3.01E+06 2.21E+06 2.94E+06 4.54E+06 3.51E+06 2.22E+06 4.26E+06 1.29E+06 1.78E+06 3.64E+06 1.65E+06 2.94E+06 4.27E+06 8.16E+05 L-Palmitoylcarnitine 3.09E+05 4.58E+05 3.39E+05 2.16E+05 1.90E+05 4.76E+05 1.24E+05 4.37E+05 4.42E+05 3.06E+05 3.04E+05 1.03E+05 1.74E+05 2.61E+05 1.69E+05 3.89E+05 8.22E+04 4.37E+05 3.58E+05 2.33E+05 6.51E+04 3.58E+04 1.74E+05 2.76E+04 1.27E+05 4.86E+05 1.35E+05 3.63E+05 2.70E+05 2.86E+04 L-Proline 2.99E+05 3.99E+05 3.47E+05 1.57E+05 1.37E+05 3.17E+05 2.51E+05 2.82E+05 4.09E+05 3.88E+05 2.34E+05 2.06E+05 2.89E+05 2.53E+05 1.97E+05 2.62E+05 2.48E+05 1.89E+05 2.12E+05 1.45E+05 1.54E+05 1.23E+05 1.48E+05 2.07E+04 9.68E+04 4.91E+05 1.21E+05 2.32E+05 4.38E+05 7.73E+04 LQ5400000 7.18E+03 4.95E+03 3.32E+03 4.22E+03 3.41E+03 7.04E+03 4.10E+03 6.12E+03 8.36E+03 2.63E+03 2.03E+03 6.62E+03 1.89E+03 3.55E+03 4.94E+03 4.19E+03 3.48E+03 4.88E+03 5.25E+03 1.50E+04 3.57E+04 4.11E+03 9.45E+03 4.98E+03 1.25E+04 1.95E+03 3.69E+03 1.49E+04 5.45E+03 1.55E+03 lys-leu 1.04E+04 1.21E+04 1.26E+04 1.13E+04 9.61E+03 1.47E+04 1.25E+04 1.65E+04 1.94E+04 1.94E+04 1.01E+04 8.31E+03 1.08E+04 8.20E+03 6.15E+03 1.03E+04 1.04E+04 6.98E+03 9.21E+03 1.15E+04 6.76E+03 4.88E+03 9.14E+03 1.77E+03 4.26E+03 1.78E+04 5.39E+03 9.85E+03 1.81E+04 3.67E+03 Methyl Jasmonate 7.63E+04 6.61E+04 5.39E+04 2.96E+04 4.42E+04 1.01E+05 5.81E+04 8.33E+04 9.61E+04 4.06E+04 2.66E+04 1.13E+05 1.44E+04 6.66E+04 7.08E+04 6.33E+04 6.08E+04 9.48E+04 8.58E+04 2.16E+05 2.23E+05 5.07E+04 7.96E+04 1.34E+05 1.68E+05 7.09E+04 4.80E+04 1.24E+05 5.95E+04 2.04E+04 MFCD00036904 3.34E+05 4.31E+05 2.85E+05 3.24E+05 2.00E+05 8.91E+04 4.46E+04 9.39E+04 1.27E+05 8.57E+04 1.20E+05 8.17E+04 1.38E+05 2.05E+04 2.77E+05 1.04E+05 5.32E+04 2.49E+05 8.71E+04 1.74E+05 1.85E+05 6.43E+04 4.95E+04 6.93E+04 1.72E+05 2.54E+05 5.60E+04 3.64E+04 1.90E+04 9.86E+03 MFCD00133435 3.32E+04 5.47E+04 3.61E+04 4.39E+04 5.72E+04 3.19E+04 1.52E+04 2.70E+04 2.22E+04 1.35E+04 3.34E+04 1.40E+04 1.74E+04 6.16E+04 3.52E+04 1.68E+04 3.37E+04 1.96E+04 2.44E+04 3.32E+04 2.16E+05 1.49E+04 2.34E+04 5.52E+04 1.80E+05 2.54E+04 1.78E+04 4.44E+04 3.87E+04 9.94E+03 MFCD02259286 4.26E+04 8.77E+04 5.61E+04 1.75E+04 2.22E+04 3.54E+05 1.63E+05 1.61E+05 2.07E+05 1.17E+05 4.55E+04 1.60E+05 3.13E+04 3.24E+05 2.95E+04 8.23E+04 1.74E+05 1.17E+05 2.38E+05 1.86E+05 3.29E+05 1.06E+05 5.96E+04 1.14E+04 5.98E+05 2.26E+05 5.94E+04 2.85E+05 1.88E+05 4.52E+04 MFCD09953737 1.27E+03 1.40E+03 1.21E+03 1.97E+05 6.79E+02 1.21E+03 1.46E+03 9.64E+02 1.83E+03 7.10E+02 4.83E+02 1.53E+03 4.75E+02 7.11E+02 1.40E+03 6.67E+02 5.79E+02 6.83E+02 7.98E+02 7.98E+03 4.90E+03 1.03E+03 1.20E+03 2.48E+03 1.94E+03 1.37E+03 5.98E+02 1.10E+03 2.87E+03 6.00E+02 N(1)-acetylspermidine 1.92E+05 2.01E+05 2.09E+05 2.41E+05 1.89E+05 2.49E+05 1.80E+05 1.54E+05 2.83E+05 2.66E+05 8.57E+04 1.18E+05 6.29E+04 1.20E+05 1.08E+05 1.05E+05 7.46E+04 4.85E+04 1.02E+05 6.23E+04 7.39E+04 5.40E+04 7.10E+04 1.70E+04 5.05E+04 1.71E+05 4.36E+04 7.05E+04 1.60E+05 3.35E+04 N-[(15Z)-tetracosenoyl]sphing-4-enine-1-phosphocholine 8.59E+04 7.37E+04 3.50E+04 1.46E+04 2.13E+04 1.08E+05 3.45E+04 1.24E+05 1.56E+05 9.29E+04 4.01E+04 3.78E+04 2.51E+04 8.66E+04 4.50E+04 8.68E+04 5.47E+04 1.15E+05 1.16E+05 1.92E+05 9.29E+04 3.17E+04 6.52E+04 1.78E+03 7.74E+04 1.74E+05 5.88E+04 4.90E+04 1.19E+05 1.46E+04 N-{3-[(4-Acetamidobutyl)amino]propyl}acetamide 1.45E+04 1.51E+04 3.88E+03 2.17E+02 1.41E+02 1.98E+04 1.13E+04 9.09E+03 1.85E+04 1.81E+04 1.12E+04 9.90E+03 7.59E+03 7.20E+03 6.73E+03 6.57E+03 4.12E+03 2.49E+03 4.92E+03 2.32E+03 1.50E+03 1.94E+03 2.00E+03 6.57E+02 1.05E+03 1.53E+04 2.12E+03 5.24E+03 1.15E+04 2.20E+03 PC 2.00E+05 2.10E+05 9.81E+04 5.00E+04 6.92E+04 1.42E+05 9.08E+04 1.92E+05 2.29E+05 1.73E+05 9.12E+04 8.01E+04 4.67E+04 3.35E+04 2.63E+05 1.71E+05 9.70E+04 2.30E+05 1.74E+05 2.56E+05 1.15E+05 1.80E+04 6.00E+04 7.51E+02 1.52E+05 1.83E+05 7.74E+04 2.51E+05 9.16E+04 5.46E+04 PEG-4 2.28E+05 2.80E+05 1.34E+05 7.67E+04 1.38E+05 4.03E+05 1.31E+05 2.46E+05 1.66E+05 1.36E+05 6.48E+04 3.66E+05 4.37E+04 1.65E+05 1.95E+05 1.28E+05 1.66E+05 2.09E+05 2.26E+05 6.64E+05 1.03E+06 1.19E+05 2.13E+05 4.38E+05 1.45E+06 1.90E+05 7.91E+04 2.61E+05 1.48E+05 8.34E+04 Platelet-activating factor 1.91E+05 2.51E+05 1.09E+05 9.46E+04 9.05E+04 3.55E+04 1.22E+04 5.88E+04 5.66E+04 4.54E+04 7.64E+04 2.89E+04 4.23E+04 1.60E+04 3.07E+05 6.38E+04 6.65E+03 7.21E+04 5.47E+04 1.12E+05 4.26E+04 1.11E+04 3.94E+04 5.78E+04 1.36E+05 1.39E+05 2.94E+04 9.06E+03 1.42E+04 4.90E+03 propionylcarnitine 7.25E+04 8.75E+04 6.74E+04 5.53E+04 3.46E+04 1.66E+05 1.25E+05 1.13E+05 8.60E+04 9.41E+04 4.21E+04 5.42E+04 4.83E+04 5.31E+04 3.55E+04 5.87E+04 5.58E+04 7.76E+04 6.90E+04 9.07E+04 3.64E+04 3.91E+04 7.68E+04 1.34E+04 4.88E+04 9.30E+04 3.04E+04 6.68E+04 1.50E+05 2.68E+04 Propylparaben 5.65E+04 3.05E+04 1.88E+04 7.55E+03 2.63E+04 6.29E+04 2.59E+04 3.16E+04 2.79E+04 2.06E+04 1.11E+04 3.48E+04 1.01E+04 2.89E+04 4.12E+04 4.80E+04 3.63E+04 5.00E+04 4.45E+04 8.73E+04 1.68E+05 2.43E+04 5.89E+04 6.08E+04 9.78E+04 2.11E+04 1.53E+04 5.07E+04 2.28E+04 1.04E+04 R79646H5Z8 2.42E+04 2.11E+04 1.34E+04 5.43E+03 1.12E+04 2.95E+04 2.33E+04 1.74E+04 2.44E+04 1.17E+04 9.29E+03 2.50E+04 7.42E+03 2.12E+04 2.25E+04 2.29E+04 2.87E+04 2.55E+04 1.84E+04 7.21E+04 1.24E+05 1.68E+04 2.21E+04 4.00E+04 4.89E+04 1.45E+04 9.98E+03 2.49E+04 2.43E+04 8.72E+03 RG5927200 1.78E+04 1.72E+04 1.46E+04 6.33E+03 1.00E+04 3.59E+04 1.65E+04 1.75E+04 1.85E+04 1.10E+04 7.55E+03 2.51E+04 5.83E+03 1.41E+04 1.70E+04 1.13E+04 1.58E+04 2.02E+04 1.69E+04 6.81E+04 8.72E+04 1.54E+04 2.27E+04 3.45E+04 7.15E+04 1.81E+04 1.05E+04 2.89E+04 1.87E+04 5.69E+03 Taurine 1.36E+06 1.51E+06 6.90E+05 1.46E+06 8.22E+05 2.27E+06 1.67E+06 2.08E+06 2.40E+06 2.30E+06 1.20E+06 1.85E+06 1.46E+06 1.85E+06 1.51E+06 2.21E+06 2.29E+06 1.68E+06 2.13E+06 2.82E+06 1.14E+06 8.06E+05 1.30E+06 2.65E+05 8.78E+05 3.38E+06 9.86E+05 2.23E+06 3.52E+06 6.99E+05 Tetranor 12-HETE 4.76E+04 4.28E+04 3.49E+04 1.52E+04 3.25E+04 7.00E+04 3.97E+04 4.72E+04 5.61E+04 3.17E+04 1.30E+04 5.34E+04 1.46E+04 3.79E+04 4.42E+04 3.23E+04 4.96E+04 5.47E+04 4.52E+04 1.32E+05 2.25E+05 4.42E+04 4.79E+04 8.88E+04 1.21E+05 4.08E+04 2.64E+04 6.16E+04 4.21E+04 1.55E+04 Urocanic acid 1.81E+04 8.56E+03 1.37E+04 1.05E+04 1.84E+04 1.07E+05 2.65E+04 1.84E+04 2.92E+04 5.12E+03 2.18E+03 4.70E+03 2.24E+03 6.37E+03 6.52E+03 1.71E+03 2.20E+03 4.62E+03 1.87E+04 3.71E+03 2.70E+04 4.19E+03 6.99E+03 2.78E+04 4.27E+04 3.78E+04 1.67E+03 7.21E+03 8.54E+04 3.03E+03 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Human Metabolome Database (1S,2R,5S)-2-Isopropyl-5-methylcyclohexyl 3-oxobutanoate HMDB0032369 (2E)-decenoic acid HMDB0010726 Dihydroartemisinin HMDB0060593 (5E,9Z)-farnesyl acetone HMDB0034495 (R)-3-hydroxybutyrylcarnitine HMDB0062735 15,16-DiHODE HMDB0010208 1-Linoleoyl-2-Hydroxy-sn-glycero-3-PC HMDB0061692 1-Methylhistamine HMDB0000898 1-Methyl-N-(9-methyl-9-azabicyclo[3.3.1]non-3-yl)-1H-indazole-3-carboxamide HMDB0015026 1-Stearoyl-2-hydroxy-sn-glycero-3-PE HMDB0011130 2-(Diethylamino)ethanol HMDB0033971 2,4-dihydroxyheptadec-16-ynyl acetate HMDB0031048 2-[4-(3-Hydroxypropyl)-2-methoxyphenoxy]-1,3-propanediol HMDB0040352 2'-Deoxycytidine HMDB0000014 2-Methoxypyrazine HMDB0033156 2-Octenoic acid HMDB0001568 3,4-Dihydroxyphenylglycol HMDB0000318 3,4-Dimethyl-5-pentyl-2-furanpropanoic acid HMDB0112083 3-[(3-Methylbutyl)nitrosoamino]-2-butanone HMDB0033553 4-[(2-Isopropyl-5-methylcyclohexyl)oxy]-4-oxobutanoic acid HMDB0036143 4-Aminobenzoic acid HMDB0001392 4-Hydroxy-5-methoxy-4-[2-methyl-3-(3-methyl-2-buten-1-yl)-2-oxiranyl]-1-oxaspiro[2.5]octan-6-one HMDB0038120 5,5'-[Disulfanediylbis({3-[(carboxymethyl)amino]-3-oxo-1,2-propanediyl}imino)]bis(2-amino-5-oxopentanoic acid) HMDB0003337 5-Isobutyl-2-oxotetrahydro-3-furancarboxylic acid HMDB0030992 6-Methoxyelemicin HMDB0034046 7-Hydroxymyoporone HMDB0035210 Acetylcarnitine HMDB0000201 Adenosine monophosphate HMDB0000045 Alpha-GPC HMDB0000086 Aminolevulinic acid HMDB0001149 Butenylcarnitine HMDB0013126 Butylparaben HMDB0032575 C14-Carnitine HMDB0254979 C4-Carnitine HMDB0002013 cis-3-Hexenyl Pyruvate HMDB0038276 Colfosceril palmitate HMDB0000564 Cyclohexylamine HMDB0031404 Cytosine HMDB0000630 Deanol HMDB0032231 Diethylamine HMDB0041878 DL-Citrulline HMDB0000904 DL-Glutamic acid HMDB0060475 D-PANTOTHENIC ACID HMDB0000210 Ethylenediaminetetraacetic acid HMDB0015109 g-Butyrobetaine HMDB0006831 HEPES HMDB0062295 hercynine HMDB0029422 Hexylresorcinol HMDB0032567 His-pro HMDB0028893 Histamine HMDB0000870 Inosinic acid HMDB0000175 Inspra HMDB0014838 L-(-)-Serine HMDB0000187 L-(+)-Arginine HMDB0000517 L-(+)-Aspartic acid HMDB0000191 L-(+)-ERGOTHIONEINE HMDB0003045 L-(+)-glutamine HMDB0000641 L-(+)-Leucine HMDB0000687 L-(+)-Valine HMDB0000883 L-Palmitoylcarnitine HMDB0000222 L-Proline HMDB0000162 LQ5400000 HMDB0041485 lys-leu HMDB0028955 Methyl Jasmonate HMDB0036583 MFCD00036904 HMDB0010382 MFCD00133435 HMDB0002815 MFCD02259286 HMDB0010168 MFCD09953737 HMDB0000953 N(1)-acetylspermidine HMDB0001276 N-[(15Z)-tetracosenoyl]sphing-4-enine-1-phosphocholine HMDB0011696 N-{3-[(4-Acetamidobutyl)amino]propyl}acetamide HMDB0041947 PC HMDB0062690 PEG-4 HMDB0094708 Platelet-activating factor HMDB0062195 propionylcarnitine HMDB0000824 Propylparaben HMDB0032574 R79646H5Z8 HMDB0240222 RG5927200 HMDB0030428 Taurine HMDB0000251 Tetranor 12-HETE HMDB0060055 Urocanic acid HMDB0000301 METABOLITES_END #END