Summary of Study ST000314

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR000253. The data can be accessed directly via it's Project DOI: 10.21228/M8302K This work is supported by NIH grant, U2C- DK119886.

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Study ID	ST000314
Study Title	NSAID treatment alters the metabolomics profile of liver, kidney, lung, and heart in an experimental mouse model of heat stroke
Study Type	Metabolomics
Study Summary	The objective of this study is to exploit broad spectrum metabolomic analysis to identify new biomarkers of multi-organ damage that will improve heat stroke (HS) diagnosis and treatment. The central hypothesis is that HS will lead to significant alterations in multi-organ metabolomics profiles that will serve as markers of HS severity, which will be shifted and intensified further by the acute use of NSAIDs. To test this hypothesis, we will be performing broad spectrum metabolomics to identify alternations in the metabolic signatures of key organs (heart, liver, kidney, and lung) in a highly validated rodent HS model leveraging implantable radiotelemetry. We will then compare these results with already completed histological gene/protein expression analysis to determine the best metabolic markers of HS induced organ damage. The results from this study will aid in the identification of preventative measures to reduce HS risk, as well as in developing therapeutics to treat multi-organ damage and facilitate recovery. The proposed study will provide the first metabolic assessment of HS severity and NSAID use, which will support future studies in HS patients to validate novel biomarkers that will improve clinical assessment of organ damage and recovery.
Institute	University of North Carolina
Department	Systems and Translational Sciences
Laboratory	Sumner Lab
Last Name	Sumner
First Name	Susan
Address	Eastern Regional Comprehensive Metabolomics Resource Core, UNC Nutrition Research Institute, 500 Laureate Way, Kannapolis, NC, 28081
Email	susan_sumner @unc.edu
Phone	704-250-5066
Submit Date	2015-12-31
Num Groups	83
Raw Data Available	Yes
Raw Data File Type(s)	fid
Analysis Type Detail	NMR
Release Date	2016-12-31
Release Version	1

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Sample Preparation:

Sampleprep ID:	SP000342
Sampleprep Summary:	Aliquots of liver sample were shipped to the NIH RTI-RCMRC on dry ice and immediately stored at -70 °C after being logged in for metabolomics analysis. Frozen tissue samples were transferred to labeled tubes containing stainless steel homogenization beads on dry ice to confirm weights. A total of 83 study samples were thawed on ice for sample preparation. Total tissue contents were extracted during homogenization with a 50:50 acetonitrile:water solution, to generate 2.0-2.5 mg/µl sample homogenates. Samples were centrifuged and an 80 mg equivalent volume (400/320 µl) of homogenized liver supernatants were transferred to new 2.0 mL tubes for the experimental samples. Analytical quality control (QC) phenotypic pool samples were generated by transferring a 50/40 µL aliquot of each supernatant from each respective phenotypic group based on HS-exposure or Controls into different 5 mL tubes. The two phenotypic QC pooled samples were vortexed to mix and a total study QC pool was generated by transferring 650 µL aliquots of each phenotypic pool sample into a new 5 mL tube for mixing. All pooled samples were aliquoted into labeled 2.0 mL tubes like the experimental samples, three for each QC group for an additional 9 samples. The samples were frozen for 2 hr at -70 °C and lyophilized to dryness overnight. Samples were reconstituted in 700 µl of D2O master mix containing 0.2 M Phosphate buffer, pH 7.4 + Chenomx ISTD with 6 mM Imidazole, 9:1 v/v. The tubes were vortexed for 2 min on a multi-tube vortexer and centrifuged at 16,000 rcf for 4 min. A 600 µl aliquot of the supernatant was transferred into pre-labeled 5 mm (4”) NMR tubes for data acquisition on a 700 MHz spectrometer.

Sampleprep ID:

SP000342

Sampleprep Summary:

Aliquots of liver sample were shipped to the NIH RTI-RCMRC on dry ice and immediately stored at -70 °C after being logged in for metabolomics analysis. Frozen tissue samples were transferred to labeled tubes containing stainless steel homogenization beads on dry ice to confirm weights. A total of 83 study samples were thawed on ice for sample preparation. Total tissue contents were extracted during homogenization with a 50:50 acetonitrile:water solution, to generate 2.0-2.5 mg/µl sample homogenates. Samples were centrifuged and an 80 mg equivalent volume (400/320 µl) of homogenized liver supernatants were transferred to new 2.0 mL tubes for the experimental samples. Analytical quality control (QC) phenotypic pool samples were generated by transferring a 50/40 µL aliquot of each supernatant from each respective phenotypic group based on HS-exposure or Controls into different 5 mL tubes. The two phenotypic QC pooled samples were vortexed to mix and a total study QC pool was generated by transferring 650 µL aliquots of each phenotypic pool sample into a new 5 mL tube for mixing. All pooled samples were aliquoted into labeled 2.0 mL tubes like the experimental samples, three for each QC group for an additional 9 samples. The samples were frozen for 2 hr at -70 °C and lyophilized to dryness overnight. Samples were reconstituted in 700 µl of D2O master mix containing 0.2 M Phosphate buffer, pH 7.4 + Chenomx ISTD with 6 mM Imidazole, 9:1 v/v. The tubes were vortexed for 2 min on a multi-tube vortexer and centrifuged at 16,000 rcf for 4 min. A 600 µl aliquot of the supernatant was transferred into pre-labeled 5 mm (4”) NMR tubes for data acquisition on a 700 MHz spectrometer.