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MB Sample ID: SA255650
Local Sample ID: | Uninfected-6 |
Subject ID: | SU002645 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU002645 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
Uninfected-6 | SA255650 | FL032734 | Uninfected | Factor |
Collection:
Collection ID: | CO002638 |
Collection Summary: | At 7 weeks post-infection, isoflurane-anesthetized mice were retro-orbitally bled and sera was flash frozen and sent to the National Institute of Health (NIH) West Coast Metabolomics Center (UC Davis) for untargeted mass spectrometry analysisusing the primary metabolism assay (ALEXCIS GCTOF-MS) or the complex lipids (CSH-QTOF MS) assay. Detected meatbolites were identified based on retention time and mass spectra from MassBank of North America, curated by the NIH West Coast Metabolomics Center , and reported as raw peak heights. The raw peak heights from each analytical platform were normalized to the average peak heights of the identified metabolites in uninfected group. The resulting data were analyzed for fold-change and multiple unpaired t-test and visualized using volcano plots to identify the differential expression of metabolites in response to T. gondii-induced cachexia |
Sample Type: | Blood (serum) |
Treatment:
Treatment ID: | TR002657 |
Treatment Summary: | To generate cysts for infection, 8-10 week female CBA/J mice were infected with 10 Me49 bradyzoite cysts by intraperitoneal injection. 4–8 weeks following infection, mice were euthanized by CO2 inhalation and cysts were harvest from brains homogenate passed through a 70 μm filter. Homegenate was washed 3 times in PBS, stained with dolichos biflorus agglutinin conjugated to FITC (Vector labs) at a 1:500 dilution. The number of cysts were determined by counting FITCpositive cysts at 20x magnification using an EVOS FL imaging system (Thermo Fisher). For experimental infections 10–14-week-old male C57BL/6 mice were infected with 10 Me49 bradyzoite cysts by intraperitoneal infection resuspended in 200 Μl PBS per mouse using a 5G 5/8” tuberculin syringe. Prior to infection, mice were cross-housed on dirty, wood chip bedding for two weeks to normalize commensal microbiota and limit the effect of eating corn husk bedding on dietary metabolites. At experimental endpoints, mice were fasted for 4 hours and isoflurane anaesthetized to isolate sera via retro-orbital bleed and/or euthanized by CO2 asphyxiation to harvest tissues for weighing and histological analysis. |
Sample Preparation:
Sampleprep ID: | SP002651 |
Sampleprep Summary: | At 7 weeks post-infection, isoflurane-anesthetized mice were retro-orbitally bled and sera was flash frozen and sent to the National Institute of Health (NIH) West Coast Metabolomics Center (UC Davis) for untargeted mass spectrometry analysisusing the primary metabolism assay (ALEXCIS GCTOF-MS) or the complex lipids (CSH-QTOF MS) assay. Detected meatbolites were identified based on retention time and mass spectra from MassBank of North America, curated by the NIH West Coast Metabolomics Center , and reported as raw peak heights (Table S1 and S3). The raw peak heights from each analytical platform were normalized to the average peak heights of the identified metabolites in uninfected group. The resulting data were analyzed for fold-change and multiple unpaired t-test and visualized using volcano plots to identify the differential expression of metabolites in response to T. gondii-induced cachexia |
Combined analysis:
Analysis ID | AN004192 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Agilent 1290 Infinity |
Column | Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um) |
MS Type | ESI |
MS instrument type | QTOF |
MS instrument name | Agilent 6530 QTOF |
Ion Mode | POSITIVE |
Units | reads |
Chromatography:
Chromatography ID: | CH003106 |
Instrument Name: | Agilent 1290 Infinity |
Column Name: | Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um) |
Column Temperature: | 65 |
Flow Gradient: | 15%-99% |
Flow Rate: | 0.6 ml/min |
Solvent A: | 60% water/40% acetonitrile; 5 mM ammonium formate; 0.1% formic acid |
Solvent B: | 90% isopropanol/10% methanol; 5 mM ammonium formate; 0.1% formic acid |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS003939 |
Analysis ID: | AN004192 |
Instrument Name: | Agilent 6530 QTOF |
Instrument Type: | QTOF |
MS Type: | ESI |
MS Comments: | Samples extracted using Matyash extraction procedure which includes MTBE, MeOH, and H2O. The organic (upper) phase was dried down and submitted for resuspension and injection onto the LC while the aqueous (bottom) phase was dried down and submitted to derivatization for GC. They are resuspended with 110 uL of a solution of 9:1 methanol: toluene and 50 ng/mL CUDA. This is then shaken for 20 seconds, sonicated for 5 minutes at room temperature, and then centrifuged for 2 minutes at 16100 rcf. The samples are then aliquoted into three parts. 33 uL are aliquoted into a vial with a 50 uL glass insert for positive and negative mode lipidomics. The last part is aliquoted into an eppendorf tube to be used as a pool. The samples are then loaded up on an Agilent 1290 Infinity LC stack. The positive and negative mode were run on an Agilent 6530 with a scan range of m/z 120-1200 Da with an acquisition speed of 2 spectra/s. Between 0.5 and 2 uL were injected onto a Waters ACQUITY UPLC CSH C18 1.7um 2.1x100mm Column with VanGuard PreColumn 2.1x5 mm . The gradient used is 0 min 15% (B), 0–2 min 30% (B), 2–2.5 min 48% (B), 2.5–11 min 82% (B), 11–11.5 min 99% (B), 11.5–12 min 99% (B), 12–12.1 min 15% (B), 12.1–15 min 15% (B) with a flow rate of 0.6 mL/min. The mass resolution for the Agilent 6530 is 10,000 for ESI (+). |
Ion Mode: | POSITIVE |