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MB Sample ID: SA014163
| Local Sample ID: | L_12_268 |
| Subject ID: | SU000334 |
| Subject Type: | Animal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57/BL-6 |
| Gender: | Male |
| Species Group: | Mammal |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU000334 |
| Subject Type: | Animal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57/BL-6 |
| Gender: | Male |
| Species Group: | Mammal |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| L_12_268 | SA014163 | FL003267 | Heat Stroke | heat_group |
| L_12_268 | SA014163 | FL003267 | T2 | time |
| L_12_268 | SA014163 | FL003267 | 125 ug INDO | drug_group |
Collection:
| Collection ID: | CO000328 |
| Collection Summary: | - |
| Sample Type: | liver tissue |
| Storage Conditions: | -70C |
Treatment:
| Treatment ID: | TR000348 |
| Treatment Summary: | Mouse heat stroke model accurately simulates the thermoregulatory, inflammatory, and organ damage responses observed in HS patients. Mice were Surgically implanted with radiotelemetry devices for the continual recording of core temperatrue during heat expsoure and 24 hours of HS recorvery. Mice were orally trated with vehicle and no drug (bacon-flavored treat) or indomethacin (5 mg/kg contained in a bacon-flavored treat) immediately prior to expsoure to a heated chamger (Model 3950, Therma Forma, Marietta, OH; ambient temperature; = 39.5 degrees celsius). Mice remained in the heated chamber, in the absence of food and water, until a maximum of 42.4 degrees celsius was reached. At the maximum temperature mice were removed from the heat and either sacrificed or provided ad libitum food and water until sacrifice at ~3 hours (when mice displayed maximum hypothermia depth; ~30 degrees C) or 24 hours of recovery (24 hrs after the start of heat exposure) when ~1.0 C fever was displayed. Heart, liver, lung, and kidney were rapidly excised, sliced into transverse or longitudinal sections, and fixed in 10% neutral-buffered formalin for histological analysis (Carson Millonig Formulation, Fisher Scientific, Springfield, MA) or stored at -80 degrees celcius for molecular and metabolomics analysis. |
| Treatment Protocol ID: | USARIEM Protocol A1002 - NSAIDS (INDO) |
| Treatment Compound: | Indomethacin |
| Treatment Route: | Oral |
| Treatment Dose: | 5 mg/kg |
| Treatment Vehicle: | Bacon-flavored treat |
| Animal Endp Tissue Coll List: | Heart, Liver, lungs and Kidneys |
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. |
Analysis:
| MB Sample ID: | SA014163 |
| Analysis ID: | AN000500 |
| Laboratory Name: | David H. Murdock Research Institute. |
| Analysis Type: | NMR |
| Software Version: | TopSpin 3.2 |
| Operator Name: | Kevin Knagge |
| Data Format: | fid, 1r |
| Num Factors: | 15 |
NMR:
| NMR ID: | NM000062 |
| Analysis ID: | AN000500 |
| Instrument Name: | Bruker |
| Instrument Type: | FT-NMR |
| NMR Experiment Type: | Other |
| Field Frequency Lock: | Deuterium |
| Standard Concentration: | 0.5 mM |
| Spectrometer Frequency: | 700 MHz |
| NMR Probe: | 5 mm ATMA Cryoprobe |
| NMR Solvent: | D2O |
| NMR Tube Size: | 5mm, 4 inch |
| Shimming Method: | Topshim |
| Pulse Sequence: | noesypr1d |
| Water Suppression: | yes |
| Receiver Gain: | 4.5 |
| Offset Frequency: | 3299.5 |
| Chemical Shift Ref Cpd: | DSS |
| Temperature: | 298.1 K |
| Number Of Scans: | 128 |
| Dummy Scans: | 4 |
| Acquisition Time: | 3.893 |
| Spectral Width: | 12.0227 ppm, 8.417 Hz |
| Num Data Points Acquired: | 65536 |
| Real Data Points: | 65536 |
| Line Broadening: | 0.5 Hz |
| Zero Filling: | yes |
| Apodization: | Lorentzian |
| Baseline Correction Method: | Polynomial |
| Chemical Shift Ref Std: | DSS-D6 |
| Binned Increment: | 0.04 |
| Binned Data Excluded Range: | 4.755 - 4.855 (water); 7.20 - 7.35 (Imidazole) |
