{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002178","ANALYSIS_ID":"AN003566","VERSION":"1","CREATED_ON":"May 27, 2022, 12:33 pm"},

"PROJECT":{"PROJECT_TITLE":"Age-independent Cardiac Protection by Pharmacological Activation of Beclin-1 During Endotoxemia and Its Association with Energy Metabolic Reprograming in Myocardium — A Targeted Metabolomics Study","PROJECT_TYPE":"A Targeted Metabolomics Study","PROJECT_SUMMARY":"Background: We previously showed that Beclin-1-dependent autophagy is cardiac protective in a rodent model of endotoxemia using young adult mice. In this report, we compared the potential therapeutic effects of pharmacological Beclin-1 activating peptide, TB-peptide, on the cardiac outcomes of young adult and aged mice during endotoxemia. We further examined alterations in myocardial metabolism induced by lipopolysaccharide (LPS) challenge with and without the TB-peptide treatment. Methods and Results: C57BL/6J mice of 10-week and 24-month-old were challenged by LPS at doses at which cardiac dysfunction occurred. Following the treatment of TB-peptide or control vehicle, heart contractility, circulating cytokines, and myocardial autophagy were evaluated. A targeted metabolomics assay was applied to analyze cardiac metabolism. TB-peptide boosted autophagic response, attenuated cytokine production, and improved cardiac performance in both young and aged mice during endotoxemia. A targeted metabolomics assay was designed to detect a pool of 361 known metabolites, of which 156 were detected in at least one of the heart tissue samples. LPS-induced impairments were found in glucose and amino acid (AA) metabolisms in mice of all ages, and TB-peptide provided ameliorative effects to rescue these alterations. However, lipid metabolites were upregulated in the young group but moderately downregulated in the aged by LPS, suggesting an age-dependent response. TB-peptide mitigated LPS-mediated trend of lipids in the young mice but provided little effect on the aged ones. Conclusion: Pharmacological activation of Beclin-1 by TB-peptide protects the heart in both young and aged population during endotoxemia, suggest a therapeutic potential for sepsis-induced cardiomyopathy. Metabolomics analysis suggests that this age-independent protection by TB-peptide is associated with reprograming of energy production via glucose and AA metabolisms.","INSTITUTE":"Loyola University Chicago Stritch School of Medicine","DEPARTMENT":"Surgery","LAST_NAME":"Zang","FIRST_NAME":"Qun","ADDRESS":"2160 S. 1st Ave, Maywood, IL 60153","EMAIL":"qzang@luc.edu","PHONE":"708-327-2472","FUNDING_SOURCE":"This work is supported by Nathan Shock Center Pilot Award (to Q.S.Z.), National Institute of Health grant 2R01GM111295-01 (to Q.S.Z.), HL109471 and CA215063 (to Z.P.L.), R01HL158515 and R01GM124108 (to J.L.), R01AG049494 (to D.P), American Heart Association grant AHA 19TP34910172 (to Z.P.L., AND NIH P30 AG013280 (to the University of Washington Nathan Shock Center). We also acknowledge The Northwest Metabolomics Research Center at the University of Washington, Seattle, and NIH S10 Grant 1S10OD021562-01 (to D.R) that funded a purchase of the LC-MS system used to acquire targeted metabolomics data.","CONTRIBUTORS":"Matthew Kim, B.S, Azadeh Nikouee, PhD, Raymond Zou, Di Ren, PhD, Zhibin He, PhD, Ji Li, PhD, Lu Wang, PhD, Danijel Djukovic, PhD, Daniel Raftery, PhD, Hayley Purcell, B.S, Daniel Promislow, PhD, Yuxiao Sun, PhD, Mohammad Goodarzi, PhD, Qing-Jun Zhang, PhD, Zhi-Ping Liu, PhD, and Qun Sophia Zang, PhD"},

"STUDY":{"STUDY_TITLE":"Age-independent Cardiac Protection by Pharmacological Activation of Beclin-1 During Endotoxemia and Its Association with Energy Metabolic Reprograming in Myocardium — A Targeted Metabolomics Study","STUDY_SUMMARY":"Background: We previously showed that Beclin-1-dependent autophagy is cardiac protective in a rodent model of endotoxemia using young adult mice. In this report, we compared the potential therapeutic effects of pharmacological Beclin-1 activating peptide, TB-peptide, on the cardiac outcomes of young adult and aged mice during endotoxemia. We further examined alterations in myocardial metabolism induced by lipopolysaccharide (LPS) challenge with and without the TB-peptide treatment. Methods and Results: C57BL/6J mice of 10-week and 24-month-old were challenged by LPS at doses at which cardiac dysfunction occurred. Following the treatment of TB-peptide or control vehicle, heart contractility, circulating cytokines, and myocardial autophagy were evaluated. A targeted metabolomics assay was applied to analyze cardiac metabolism. TB-peptide boosted autophagic response, attenuated cytokine production, and improved cardiac performance in both young and aged mice during endotoxemia. A targeted metabolomics assay was designed to detect a pool of 361 known metabolites, of which 156 were detected in at least one of the heart tissue samples. LPS-induced impairments were found in glucose and amino acid (AA) metabolisms in mice of all ages, and TB-peptide provided ameliorative effects to rescue these alterations. However, lipid metabolites were upregulated in the young group but moderately downregulated in the aged by LPS, suggesting an age-dependent response. TB-peptide mitigated LPS-mediated trend of lipids in the young mice but provided little effect on the aged ones. Conclusion: Pharmacological activation of Beclin-1 by TB-peptide protects the heart in both young and aged population during endotoxemia, suggest a therapeutic potential for sepsis-induced cardiomyopathy. Metabolomics analysis suggests that this age-independent protection by TB-peptide is associated with reprograming of energy production via glucose and AA metabolisms.","INSTITUTE":"Loyola University Chicago Stritch School of Medicine","DEPARTMENT":"Surgery","LAST_NAME":"Zang","FIRST_NAME":"Qun","ADDRESS":"2160 S. 1st Ave, Maywood, IL 60153","EMAIL":"qzang@luc.edu","PHONE":"708-327-2472"},

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"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-7_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-8_NEG",
"Factors":{"Age Group":"Old","Treatment":"peptide"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-8_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-9_NEG",
"Factors":{"Age Group":"Young","Treatment":"Ctrl"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-9_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-10_NEG",
"Factors":{"Age Group":"Old","Treatment":"LPS1-peptide"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-10_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-QC(I)#2_NEG",
"Factors":{"Age Group":"instrument quality control","Treatment":"-"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-QC(I)#2_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-QC(S)#2_NEG",
"Factors":{"Age Group":"sample quality control","Treatment":"-"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-QC(S)#2_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-11_NEG",
"Factors":{"Age Group":"Young","Treatment":"LPS3"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-11_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-12_NEG",
"Factors":{"Age Group":"Young","Treatment":"LPS3_peptide"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-12_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-13_NEG",
"Factors":{"Age Group":"Young","Treatment":"LPS3_peptide"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-13_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-14_NEG",
"Factors":{"Age Group":"Young","Treatment":"LPS3"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-14_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-15_NEG",
"Factors":{"Age Group":"Young","Treatment":"LPS3_peptide"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-15_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-16_NEG",
"Factors":{"Age Group":"Old","Treatment":"LPS1_peptide"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-16_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-17_NEG",
"Factors":{"Age Group":"Young","Treatment":"Ctrl"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-17_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-18_NEG",
"Factors":{"Age Group":"Young","Treatment":"peptide"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-18_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-19_NEG",
"Factors":{"Age Group":"Young","Treatment":"Ctrl"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-19_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-20_NEG",
"Factors":{"Age Group":"Young","Treatment":"LPS3_peptide"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-20_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-QC(I)#3_NEG",
"Factors":{"Age Group":"instrument quality control","Treatment":"-"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-QC(I)#3_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-QC(S)#3_NEG",
"Factors":{"Age Group":"sample quality control","Treatment":"-"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-QC(S)#3_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-21_NEG",
"Factors":{"Age Group":"Old","Treatment":"Ctrl"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-21_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-22_NEG",
"Factors":{"Age Group":"Young","Treatment":"peptide"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-22_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-23_NEG",
"Factors":{"Age Group":"Old","Treatment":"Ctrl"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-23_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-24_NEG",
"Factors":{"Age Group":"Young","Treatment":"LPS3_peptide"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-24_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-25_NEG",
"Factors":{"Age Group":"Young","Treatment":"LPS3"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-25_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-26_NEG",
"Factors":{"Age Group":"Old","Treatment":"LPS1"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-26_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-27_NEG",
"Factors":{"Age Group":"Old","Treatment":"LPS1-peptide"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-27_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-28_NEG",
"Factors":{"Age Group":"Old","Treatment":"Ctrl"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-28_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-29_NEG",
"Factors":{"Age Group":"Old","Treatment":"peptide"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-29_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-30_NEG",
"Factors":{"Age Group":"Young","Treatment":"Ctrl"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-30_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-QC(I)#4_NEG",
"Factors":{"Age Group":"instrument quality control","Treatment":"-"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-QC(I)#4_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-QC(S)#4_NEG",
"Factors":{"Age Group":"sample quality control","Treatment":"-"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-QC(S)#4_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-31_NEG",
"Factors":{"Age Group":"Young","Treatment":"Ctrl"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-31_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-32_NEG",
"Factors":{"Age Group":"Old","Treatment":"LPS1-peptide"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-32_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-33_NEG",
"Factors":{"Age Group":"Young","Treatment":"peptide"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-33_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-34_NEG",
"Factors":{"Age Group":"Old","Treatment":"Ctrl"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-34_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-35_NEG",
"Factors":{"Age Group":"Young","Treatment":"LPS3_peptide"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-35_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-36_NEG",
"Factors":{"Age Group":"Young","Treatment":"LPS3"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-36_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-37_NEG",
"Factors":{"Age Group":"Old","Treatment":"LPS1"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-37_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-38_NEG",
"Factors":{"Age Group":"Old","Treatment":"Ctrl"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-38_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-39_NEG",
"Factors":{"Age Group":"Old","Treatment":"LPS1-peptide"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-39_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-40_NEG",
"Factors":{"Age Group":"Old","Treatment":"LPS1"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-40_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-41_NEG",
"Factors":{"Age Group":"Young","Treatment":"peptide"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-41_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-42_NEG",
"Factors":{"Age Group":"Young","Treatment":"peptide"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-42_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-43_NEG",
"Factors":{"Age Group":"Old","Treatment":"peptide"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-43_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-QC(I)#5_NEG",
"Factors":{"Age Group":"instrument quality control","Treatment":"-"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-QC(I)#5_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-QC(S)#5_NEG",
"Factors":{"Age Group":"sample quality control","Treatment":"-"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-QC(S)#5_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-Blank0_NEG",
"Factors":{"Age Group":"blank","Treatment":"-"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-Blank0_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-Blank_NEG",
"Factors":{"Age Group":"blank","Treatment":"-"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-Blank_NEG.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-Blank_NEG (2)",
"Factors":{"Age Group":"blank","Treatment":"-"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-Blank_NEG (2).mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-Blank_NEG (3)",
"Factors":{"Age Group":"blank","Treatment":"-"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-Blank_NEG (3).mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-Blank_NEG (4)",
"Factors":{"Age Group":"blank","Treatment":"-"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-Blank_NEG (4).mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-Blank_NEG (5)",
"Factors":{"Age Group":"blank","Treatment":"-"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-Blank_NEG (5).mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-Blank_NEG (6)",
"Factors":{"Age Group":"blank","Treatment":"-"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-Blank_NEG (6).mzML"}
},
{
"Subject ID":"-",
"Sample ID":"2020-08-17_Zang-43_Box2Redo-Dummy_NEG",
"Factors":{"Age Group":"instrument quality control","Treatment":"-"},
"Additional sample data":{"RAW_FILE_NAME":"2020-08-17_Zang-43_Box2Redo-Dummy_NEG.mzML"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Wild-type (WT) C57BL/6 mice were used in this study. Endotoxemia was induced in young (10-week) and aged (24-week) male mice by lipopolysaccharide (LPS). Based on published results as well as observations in our laboratory, male and female mice showed significantly different susceptibility to systemic symptoms in sepsis models. Thus, male but not female mice were chosen for the experiments presented in this report. LPS was administered intraperitoneally (i.p.), and mice were weighed individually to determine the exact amount of LPS (MilliporeSigma, Burlington, MA; catalog number L3012) required to achieve the doses indicated in the figure legends. Sterile endotoxin-free PBS was used as a vehicle control in sham groups. In some experiments, Beclin-1-activating peptide (TB peptide), synthesized according to a published sequence24 by NonoPep (Shanghai, China), was administered i.p. at a dose of 16 mg/kg in 100μl of PBS 30 minutes post LPS-challenge. Heart tissue was harvested 18 hours post LPS challenge.","COLLECTION_PROTOCOL_FILENAME":"Zang_Protocol.docx","SAMPLE_TYPE":"Heart","STORAGE_CONDITIONS":"-80℃"},

"TREATMENT":{"TREATMENT_SUMMARY":"Endotoxemia was induced in young (10-week) and aged (24-week) male mice by lipopolysaccharide (LPS). Based on published results as well as observations in our laboratory, male and female mice showed significantly different susceptibility to systemic symptoms in sepsis models. Thus, male but not female mice were chosen for the experiments presented in this report. LPS was administered intraperitoneally (i.p.), and mice were weighed individually to determine the exact amount of LPS (MilliporeSigma, Burlington, MA; catalog number L3012) required to achieve the required doses. Sterile endotoxin-free PBS was used as a vehicle control in sham groups. Consistent with literature and as expected, we observed that older mice were more susceptible to the toxic effects induced by LPS. 24-month-old (aged) mice showed impaired cardiac function but were able to survive when receiving LPS challenged at 1mg/kg. However, greater fatality was observed when LPS dose was increased to 3 mg/kg. In 10-week-old (young adult) mice, 3 mg/kg LPS triggered heart dysfunction without impact on survival, whereases at 10 mg/kg, we observed significant LPS-induced fatality in the group. Due to the different sensitivities to LPS between the aged and young adult mice, we were not able to choose a universal dose of LPS to induce cardiac dysfunction and to perform follow up analysis in both groups. Therefore, we used the physiological function of the heart as a base for comparison in the studies performed in this report. Our previous research provided evidence that stimulating beclin-1 dependent autophagy improves cardiac performance during endotoxemia in young adult mice, and thus Beclin-1-activating peptide (TB peptide) holds a promising therapeutic potential for sepsis. In this report, we examined whether TB-peptide exerts a similar protective effect on aged animals under the same condition. In our experimental setting, sham or LPS challenge was administered to groups of 24-month-old and 10-week-old mice followed by treatment with TB-peptide, administered i.p. at a dose of 16 mg/kg in 100μl of PBS 30 minutes post LPS-challenge. Heart tissue was collected 18 hours post LPS challenge."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Sample preparation: aqueous metabolites were extracted using a methanol-based protein precipitation method as described previously. Briefly, heart tissue samples were homogenized in cold water using zirconium oxide beads, methanol was added, and samples were vortexed and then stored for 30 minutes at -20˚C. Afterwards, samples were first sonicated in an ice bath for 10 minutes, centrifuged for 15 min at 18,000g and 4˚C, and then a fixed volume of supernatant was collected from each sample. Lastly, recovered supernatants were dried on a SpeedVac and reconstituted for LC-MS analysis. Protein pallets that were left over from the sample prep were saved for BCA assay. (2) LC-MS analysis: samples were analyzed on a duplex-LC-MS system composed of two Shimadzu UPLC pumps, CTC Analytics PAL HTC-xt temperature-controlled auto-sampler and AB Sciex 6500+ Triple Quadrupole MS equipped with ESI ionization source. UPLC pumps were connected to the auto-sampler in parallel and were able to perform two chromatography separations independently from each other. Each sample was injected twice on two identical analytical columns (Waters XBridge BEH Amide XP) performing separations in hydrophilic interaction liquid chromatography (HILIC) mode. While one column was performing separation and MS data acquisition in ESI(+) ionization mode, the other column was getting equilibrated for sample injection, chromatography separation and MS data acquisition in ESI(-) mode. Each chromatography separation was 18 minutes (total analysis time per sample was 36 minutes)."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"HILIC","INSTRUMENT_NAME":"Shimadzu Nexera X2","COLUMN_NAME":"Waters XBridge BEH Amide 150 x 2.1mm, 2.5 um."},

"ANALYSIS":{"ANALYSIS_TYPE":"MS","ANALYSIS_PROTOCOL_FILE":"Zang_Protocol.docx"},

"MS":{"INSTRUMENT_NAME":"ABI Sciex 6500+","INSTRUMENT_TYPE":"QTRAP","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"MS data acquisition was performed in multiple-reaction-monitoring (MRM) mode. The whole LC-MS system was controlled using AB Sciex Analyst 1.6.3 software. Measured MS peaks were integrated using AB Sciex MultiQuant 3.0.3 software. In addition to the study samples, two sets of quality control (QC) samples were used to monitor the assay performance as well as data reproducibility. One QC was a pooled human serum sample used to monitor system performance and the other QC was pooled study samples and this QC was used to monitor data reproducibility. Isotope labeled compounds were used to monitor sample preparation and injection. Highly reproducible MS data were generated, having an average coefficient of variances (CVs) among the metabolites of 5.6%. Data for each sample were normalized according to bicinchoninic acid (BCA)-based quantification of total protein count."},

"MS_METABOLITE_DATA":{
"Units":"count per second (CPS)",

"Data":[{"Metabolite":"Trimethylamine 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}

}