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BCA was measured and 100ug were then reduced with 5 mM DTT at RT for 30 min and then alkylated with 10mM IAA in the dark for 30 min at RT. The samples were then digested with 4ug of Lysyl endopeptidase (Wako) overnight, diluted to 1M Urea and further digested with 4ug trypsin (Thermofisher) overnight at room temperature. The peptide solutions were then acidified to 1% FA and cleaned with 10mg HLB columns. HLB columns were conditioned with 500uL of methanol and equilibrated twice with 500uL of 1% formic acid. Samples were loaded and washed twice with 500ul of 1% formic acid. Finally the samples were eluted with 1mL of 50% acetonitrile. An aliquot of 10% was separated out and all eluates were then dried to completeness using a SpeedVac (LabConco). ###Liquid Chromatography and Mass spectrometry (Evosep ONE) Each sample was resuspended in 200 ul of loading buffer (0.1% FA) and 20 ul was loaded onto Evotips and analyzed by liquid chromatography coupled to tandem mass spectrometry. Peptide eluents were separated on IonOptick’s column (15 cm × 75 μM internal diameter (ID) packed with 1.7um resin) by a Evosep One (Evosep). Buffer A was water with 0.1% (vol/vol) formic acid, and buffer B was 100% (vol/vol) acetonitrile in water with 0.1% (vol/vol) formic acid. Elution was performed using the preset 40SPD Whisper Zoom method. Peptides were monitored on a Orbitrap Astral mass spectrometer (ThermoFisher Scientific) fitted with a high-field asymmetric waveform ion mobility spectrometry (FAIMS Pro) ion mobility source(ThermoFisher Scientific). One compensation voltages (CV) of -35 was chosen for the FAIMS. Each cycle consisted of one full scan (MS1) was performed with an m/z range of 380-980 at 240,000 resolution, 500% AGC and 3 ms injection time. The higher energy collision-induced dissociation (HCD) DIA scans were collected with a 3 m/z isolation windows over the entire precursor range (380-980 m/z) with a time of 0.6 seconds and 3 ms injection time. Collision energy was set to 27% and scan range set to 150 - 2000 m/z."],"repository":["Pride"],"modification":[""],"quantification_method":["Not available"],"data_protocol":["###Database Search Spectronaut (version 19.9.250512.62635) was used to search all rawfiles in default library free mode using a sheep uniprot database (downloaded 072024). All parameters were kept at default."],"omics_type":["Proteomics"],"labhead":["Michael Davis"],"instrument_platform":[""],"submission_type":["PARTIAL"],"labhead_affiliation":["Professor Associate Chair for Graduate Studies BME Emory GT"],"species":["Ovis Aries"],"publication":["42432461 Takaesu F, Villarreal DJ, Zhou A, Jimenez MR, Turner ME, Spiess JL, Kievert JC, DeShetler CM, Schwartzman WE, Yates AR, Kelly JM, Breuer CK, Davis ME. Serum Proteomic Profiling Implicates a Dysregulated Neurohormonal-Inflammatory Axis in Post-Fontan Sinus Tachycardia. J Am Heart Assoc. 2026 15(14):e049852 10.1161/jaha.126.049852"],"submitter_mail":["dduong@emory.edu"],"submitter_affiliation":["Center of Neurodegenerative Diseases"],"submitter_country":["United States"],"pubmed_abstract":["Background
Postoperative sinus tachycardia is a poorly understood complication following the Fontan procedure. The molecular signaling cascades triggering acute tachycardia remain uncharacterized, limiting therapeutic innovation. Here, we present a retrospective study leveraging serum proteomics and machine learning to identify the molecular drivers of postoperative Fontan sinus tachycardia.Methods
We integrated a clinically relevant ovine Fontan model with continuous telemetric heart rate monitoring and human patient data. Serum proteomics coupled with least absolute shrinkage and selection operator and Boruta machine learning algorithms were used to identify protein panels predictive of postoperative sinus tachycardia. Cross-species validation was performed by comparing proteomic signatures from sheep and pediatric patients undergoing Glenn or Fontan surgery.Results
Ovine Fontan animals demonstrated significant heart rate elevation beginning on postoperative day 1, peaking at postoperative day 3 (159.4±11.7 bpm versus preoperative, 105.3±10.5 bpm; P=0.0002), before trending toward baseline by postoperative day 10. This pattern was mirrored in human patients with a more modest magnitude. Surgical controls did not exhibit tachycardia. The principal component most correlated with heart rate (principal component 1: r=0.78, P=2.2×10-4) was enriched for inflammatory and neural pathways. The Boruta algorithm identified an 11-protein panel with strong predictive power (area under the receiver operating characteristic curve, 0.963). Cross-species comparison demonstrated that angiotensinogen, angiotensin-converting enzyme, and pentraxin 3 were similarly dysregulated in both species postoperatively.Conclusions
This study provides molecular evidence implicating a dysregulated neurohormonal-inflammatory axis in acute postoperative Fontan sinus tachycardia and establishes a foundation for developing targeted diagnostics and therapeutics for this complication."],"pubmed_title":["Serum Proteomic Profiling Implicates a Dysregulated Neurohormonal-Inflammatory Axis in Post-Fontan Sinus Tachycardia."],"pubmed_authors":["Takaesu Felipe F, Villarreal Delaney J DJ, Zhou Ashley A, Jimenez Michael R MR, Turner Mackenzie E ME, Spiess J Logan JL, Kievert Jennifer C JC, DeShetler Cameron M CM, Schwartzman William E WE, Yates Andrew R AR, Kelly John M JM, Breuer Christopher K CK, Davis Michael E ME"],"additional_accession":[]},"is_claimable":false,"name":"Felipe Sheep EV DIA LC/MS/MS using the Orbitrap Astral","description":"DIA analysis of sheep EVs using Evosep one and Orbitrap 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