Summary of Study ST001255

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 PR000841. The data can be accessed directly via it's Project DOI: 10.21228/M8011M This work is supported by NIH grant, U2C- DK119886.

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This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study IDST001255
Study TitleImmunomodulatory activity of hyaluronidase is associated with metabolic adaptations during acute inflammation
Study SummaryObjective and design: Investigate survival outcomes, and immunological and metabolomic effects of hyaluronidase (Hz) treatment during mouse models of acute inflammation and sepsis. Methods: Survival of C57Bl/6 mice was monitored after lethal challenge with lipopolysaccharide (LPS) or cecal and ligation puncture (CLP)-induced sepsis and treated with Hz or saline. Mice were also challenged with LPS and treated with Hz for leukocyte counting, cytokine quantification and determination of metabolomic profiles in the peritoneal fluid. Results: Hz treatment improved survival outcomes after lethal challenge with LPS or CLPinduced sepsis. LPS challenge promoted acute neutrophil accumulation and production of interleukin-1β (IL-1β) and IL-6 in the peritoneum, whereas Hz treatment suppressed neutrophil infiltration and cytokine production. We further characterized the metabolomic alterations caused by LPS challenge, which predicted activity of metabolic pathways related to fatty acids and eicosanoids. Hz treatment had a profound effect over the metabolic response, reflected by reductions of the relative levels of fatty acids. Conclusion: Collectively, these data demonstrate that Hz treatment is associated with metabolic reprogramming of pathways that sustain the inflammatory response.
Institute
Sao Paulo University
DepartmentSchool of Pharmaceutical Sciences of Ribeirao Preto
Last NameGardinassi
First NameLuiz Gustavo
AddressAv do Cafe, s/n - Ribeirão Preto - SP
Emailgustavogardinassi@usp.br
Phone551633154189
Submit Date2019-09-26
Num Groups8
Total Subjects40
Num Males40
Raw Data AvailableYes
Raw Data File Type(s)wiff
Analysis Type DetailLC-MS
Release Date2019-10-11
Release Version1
Luiz Gustavo Gardinassi Luiz Gustavo Gardinassi
https://dx.doi.org/10.21228/M8011M
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:

Project:

Project ID:PR000841
Project DOI:doi: 10.21228/M8011M
Project Title:Immunomodulatory activity of hyaluronidase is associated with metabolic adaptations during acute inflammation
Project Type:Untargeted MS
Project Summary:Objective and design: Investigate survival outcomes, and immunological and metabolomic effects of hyaluronidase (Hz) treatment during mouse models of acute inflammation and sepsis. Methods: Survival of C57Bl/6 mice was monitored after lethal challenge with lipopolysaccharide (LPS) or cecal and ligation puncture (CLP)-induced sepsis and treated with Hz or saline. Mice were also challenged with LPS and treated with Hz for leukocyte counting, cytokine quantification and determination of metabolomic profiles in the peritoneal fluid. Results: Hz treatment improved survival outcomes after lethal challenge with LPS or CLPinduced sepsis. LPS challenge promoted acute neutrophil accumulation and production of interleukin-1β (IL-1β) and IL-6 in the peritoneum, whereas Hz treatment suppressed neutrophil infiltration and cytokine production. We further characterized the metabolomic alterations caused by LPS challenge, which predicted activity of metabolic pathways related to fatty acids and eicosanoids. Hz treatment had a profound effect over the metabolic response, reflected by reductions of the relative levels of fatty acids. Conclusion: Collectively, these data demonstrate that Hz treatment is associated with metabolic reprogramming of pathways that sustain the inflammatory response.
Institute:Sao Paulo University
Department:Department of Clinical Analyses, Toxicology and Food Sciences
Last Name:Gardinassi
First Name:Luiz Gustavo
Address:Av do Café s/n - Ribeirão Preto - SP
Email:gustavogardinassi@usp.br
Phone:551633154189
Funding Source:FAPESP; CNPq; CAPES

Subject:

Subject ID:SU001323
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
Genotype Strain:C57Bl6
Weight Or Weight Range:22–25 g
Gender:Male
Animal Light Cycle:light/dark cycle
Animal Feed:Ad libitum
Animal Water:Ad libitum

Factors:

Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)

mb_sample_id local_sample_id Treatment Time
SA091259Hz_27h_4Hz 27h
SA091260Hz_27h_3Hz 27h
SA091261Hz_27h_2Hz 27h
SA091262Hz_27h_6Hz 27h
SA091263Hz_27h_5Hz 27h
SA091264Hz_7d_5Hz 7d
SA091265Hz_7d_1Hz 7d
SA091266Hz_7d_3Hz 7d
SA091267Hz_7d_6Hz 7d
SA091268Hz_7d_2Hz 7d
SA091269LPS_Hz_27h_1LPS + Hz 27h
SA091270LPS_Hz_27h_6LPS + Hz 27h
SA091271LPS_Hz_27h_4LPS + Hz 27h
SA091272LPS_Hz_27h_3LPS + Hz 27h
SA091273LPS_Hz_27h_2LPS + Hz 27h
SA091274LPS_Hz_27h_5LPS + Hz 27h
SA091275LPS_Hz_7d_1LPS + Hz 7d
SA091276LPS_Hz_7d_3LPS + Hz 7d
SA091277LPS_Hz_7d_2LPS + Hz 7d
SA091278LPS_27h_6LPS 27h
SA091279LPS_27h_1LPS 27h
SA091280LPS_27h_5LPS 27h
SA091281LPS_27h_3LPS 27h
SA091282LPS_27h_2LPS 27h
SA091283LPS_3h_1LPS 3h
SA091284LPS_3h_2LPS 3h
SA091285LPS_3h_5LPS 3h
SA091286LPS_3h_6LPS 3h
SA091287LPS_3h_4LPS 3h
SA091288LPS_3h_3LPS 3h
SA091289LPS_7d_6LPS 7d
SA091290LPS_7d_1LPS 7d
SA091291LPS_7d_3LPS 7d
SA091292LPS_7d_2LPS 7d
SA091293LPS_7d_4LPS 7d
SA091294LPS_7d_5LPS 7d
SA091295PBS_2PBS 0h
SA091296PBS_3PBS 0h
SA091297PBS_4PBS 0h
SA091298PBS_1PBS 0h
Showing results 1 to 40 of 40

Collection:

Collection ID:CO001317
Collection Summary:Mice were anesthetized with ketamine (100mg/kg) (Dopaser®, Minas Gerais, Brazil) and xylazine (10mg/kg) (Agener União, São Paulo, Brazil). Following euthanasia in CO2 chamber, 3mL of PBS was added into the abdominal cavity, which was gently massaged for 1 min. The peritoneal fluid was collected with a needle inserted into the inguinal region.
Sample Type:Peritoneal fluid
Storage Conditions:-80℃

Treatment:

Treatment ID:TR001338
Treatment Summary:Endotoxemia was induced in mice by intraperitoneal (i.p.) injection of LPS dissolved in 0.3 mL of phosphate-buffered saline (PBS). Control groups received sterile PBS. Tree hours (h) after LPS injection, mice were treated with 16 U of Hz (dissolved in 0.3 mL of PBS) every 8 h

Sample Preparation:

Sampleprep ID:SP001331
Sampleprep Summary:Peritoneal fluid samples were spiked with stable isotope internal standards and metabolites were enriched with solid phase extraction cartridges (Hypersep C18-500 mg, 3 mL, Thermo Scientific-Bellefonte, PA, USA) and methanol/water as solvents.

Combined analysis:

Analysis ID AN002084
Analysis type MS
Chromatography type Reversed phase
Chromatography system Shimadzu Nexera X2
Column Ascentis Express C18 (100 x 4.6mm,2.7um)
MS Type ESI
MS instrument type QTOF
MS instrument name ABI Sciex 5600 TripleTOF
Ion Mode NEGATIVE
Units peak intensity

Chromatography:

Chromatography ID:CH001521
Chromatography Summary:Reversed-phase chromatography was accomplished with a 100 × 4.6 mm, 2.7 μm Ascentis Express C18 column (Supelco - St. Louis, MO, USA) with an Ultra-High-Performance Liquid Chromatography (UHPLC) system (Nexera X2, Shimadzu-Kyoto, HO, Japan). The gradient consisted of Phase A, H2O/ACN/acetic acid (69.98:30:0.02, v/v/v) at pH 5.8 (adjusted with NH4OH), and Phase B, an ACN/isopropanol (70:30, v/v). Gradient elution was carried out for 25 min at a flow rate of 0.6 mL. / min. Gradient conditions were as follows: 0 to 2.0 min, 0% B; 2.0 to 5.0 min, 15% B; 5.0 to 8.0 min, 20% B; 8.0 to 11.0 min, 35% B; 11.0 to 15.0 min, 70% B; and 15.0 to 19 min, 100% B. At 19.0 min, the gradient returned to the initial condition of 0% B, and the column was re-equilibrated until 25.0 min.
Methods Filename:PR_CH_Methods.pdf
Instrument Name:Shimadzu Nexera X2
Column Name:Ascentis Express C18 (100 x 4.6mm,2.7um)
Flow Gradient:0 to 2.0 min, 0% B; 2.0 to 5.0 min, 15% B; 5.0 to 8.0 min, 20% B; 8.0 to 11.0 min, 35% B; 11.0 to 15.0 min, 70% B; and 15.0 to 19 min, 100% B. At 19.0 min, the gradient returned to the initial condition of 0% B, and the column was re-equilibrated until 25.0 min.
Flow Rate:0.6ml/min
Solvent A:70% water/30% acetonitrile; 0.02% acetic acid, pH 5.8(adjusted with ammonium hydroxide)
Solvent B:70% acetonitrile/30% isopropanol
Chromatography Type:Reversed phase

MS:

MS ID:MS001935
Analysis ID:AN002084
Instrument Name:ABI Sciex 5600 TripleTOF
Instrument Type:QTOF
MS Type:ESI
MS Comments:Mass spectral data was acquired with negative electrospray ionization and the full scan of mass-to-charge ratio (m/z) ranged from 100 to 1500. Proteowizard software was used to convert wiff files into mzXML files. Peak peaking, noise filtering, retention time and m/z alignment, and feature quantification were performed with apLCMS. Three parameters define a metabolite feature: mass-to-charge ratio (m/z), retention time and intensity values. Data were log2 transformed and only features detected in at least 80% of samples from one group (5439 m/z features) were used in further analysis. Missing values were imputed using half mean of the feature across all samples. The mummichog software (version 2) was used for metabolic pathway enrichment analysis (mass accuracy under 10 ppm).
Ion Mode:NEGATIVE
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