{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001255","ANALYSIS_ID":"AN002084","VERSION":"1","CREATED_ON":"September 30, 2019, 1:35 pm"},

"PROJECT":{"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, School of Pharmaceutical Sciences of Ribeirao Preto","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"},

"STUDY":{"STUDY_TITLE":"Immunomodulatory activity of hyaluronidase is associated with metabolic adaptations during acute inflammation","STUDY_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":"School of Pharmaceutical Sciences of Ribeirao Preto","LAST_NAME":"Gardinassi","FIRST_NAME":"Luiz Gustavo","ADDRESS":"Av do Cafe, s/n - Ribeirão Preto - SP","EMAIL":"gustavogardinassi@usp.br","PHONE":"551633154189","NUM_GROUPS":"8","TOTAL_SUBJECTS":"40","NUM_MALES":"40"},

"SUBJECT":{"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"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"PBS_1",
"Factors":{"Treatment":"PBS","Time":"0h"}
},
{
"Subject ID":"-",
"Sample ID":"PBS_2",
"Factors":{"Treatment":"PBS","Time":"0h"}
},
{
"Subject ID":"-",
"Sample ID":"PBS_3",
"Factors":{"Treatment":"PBS","Time":"0h"}
},
{
"Subject ID":"-",
"Sample ID":"PBS_4",
"Factors":{"Treatment":"PBS","Time":"0h"}
},
{
"Subject ID":"-",
"Sample ID":"LPS_3h_1",
"Factors":{"Treatment":"LPS","Time":"3h"}
},
{
"Subject ID":"-",
"Sample ID":"LPS_3h_2",
"Factors":{"Treatment":"LPS","Time":"3h"}
},
{
"Subject ID":"-",
"Sample ID":"LPS_3h_3",
"Factors":{"Treatment":"LPS","Time":"3h"}
},
{
"Subject ID":"-",
"Sample ID":"LPS_3h_4",
"Factors":{"Treatment":"LPS","Time":"3h"}
},
{
"Subject ID":"-",
"Sample ID":"LPS_3h_5",
"Factors":{"Treatment":"LPS","Time":"3h"}
},
{
"Subject ID":"-",
"Sample ID":"LPS_3h_6",
"Factors":{"Treatment":"LPS","Time":"3h"}
},
{
"Subject ID":"-",
"Sample ID":"LPS_27h_1",
"Factors":{"Treatment":"LPS","Time":"27h"}
},
{
"Subject ID":"-",
"Sample ID":"LPS_27h_2",
"Factors":{"Treatment":"LPS","Time":"27h"}
},
{
"Subject ID":"-",
"Sample ID":"LPS_27h_3",
"Factors":{"Treatment":"LPS","Time":"27h"}
},
{
"Subject ID":"-",
"Sample ID":"LPS_27h_5",
"Factors":{"Treatment":"LPS","Time":"27h"}
},
{
"Subject ID":"-",
"Sample ID":"LPS_27h_6",
"Factors":{"Treatment":"LPS","Time":"27h"}
},
{
"Subject ID":"-",
"Sample ID":"LPS_7d_1",
"Factors":{"Treatment":"LPS","Time":"7d"}
},
{
"Subject ID":"-",
"Sample ID":"LPS_7d_2",
"Factors":{"Treatment":"LPS","Time":"7d"}
},
{
"Subject ID":"-",
"Sample ID":"LPS_7d_3",
"Factors":{"Treatment":"LPS","Time":"7d"}
},
{
"Subject ID":"-",
"Sample ID":"LPS_7d_4",
"Factors":{"Treatment":"LPS","Time":"7d"}
},
{
"Subject ID":"-",
"Sample ID":"LPS_7d_5",
"Factors":{"Treatment":"LPS","Time":"7d"}
},
{
"Subject ID":"-",
"Sample ID":"LPS_7d_6",
"Factors":{"Treatment":"LPS","Time":"7d"}
},
{
"Subject ID":"-",
"Sample ID":"Hz_27h_2",
"Factors":{"Treatment":"Hz","Time":"27h"}
},
{
"Subject ID":"-",
"Sample ID":"Hz_27h_3",
"Factors":{"Treatment":"Hz","Time":"27h"}
},
{
"Subject ID":"-",
"Sample ID":"Hz_27h_4",
"Factors":{"Treatment":"Hz","Time":"27h"}
},
{
"Subject ID":"-",
"Sample ID":"Hz_27h_5",
"Factors":{"Treatment":"Hz","Time":"27h"}
},
{
"Subject ID":"-",
"Sample ID":"Hz_27h_6",
"Factors":{"Treatment":"Hz","Time":"27h"}
},
{
"Subject ID":"-",
"Sample ID":"Hz_7d_1",
"Factors":{"Treatment":"Hz","Time":"7d"}
},
{
"Subject ID":"-",
"Sample ID":"Hz_7d_2",
"Factors":{"Treatment":"Hz","Time":"7d"}
},
{
"Subject ID":"-",
"Sample ID":"Hz_7d_3",
"Factors":{"Treatment":"Hz","Time":"7d"}
},
{
"Subject ID":"-",
"Sample ID":"Hz_7d_5",
"Factors":{"Treatment":"Hz","Time":"7d"}
},
{
"Subject ID":"-",
"Sample ID":"Hz_7d_6",
"Factors":{"Treatment":"Hz","Time":"7d"}
},
{
"Subject ID":"-",
"Sample ID":"LPS_Hz_27h_1",
"Factors":{"Treatment":"LPS + Hz","Time":"27h"}
},
{
"Subject ID":"-",
"Sample ID":"LPS_Hz_27h_2",
"Factors":{"Treatment":"LPS + Hz","Time":"27h"}
},
{
"Subject ID":"-",
"Sample ID":"LPS_Hz_27h_3",
"Factors":{"Treatment":"LPS + Hz","Time":"27h"}
},
{
"Subject ID":"-",
"Sample ID":"LPS_Hz_27h_4",
"Factors":{"Treatment":"LPS + Hz","Time":"27h"}
},
{
"Subject ID":"-",
"Sample ID":"LPS_Hz_27h_5",
"Factors":{"Treatment":"LPS + Hz","Time":"27h"}
},
{
"Subject ID":"-",
"Sample ID":"LPS_Hz_27h_6",
"Factors":{"Treatment":"LPS + Hz","Time":"27h"}
},
{
"Subject ID":"-",
"Sample ID":"LPS_Hz_7d_1",
"Factors":{"Treatment":"LPS + Hz","Time":"7d"}
},
{
"Subject ID":"-",
"Sample ID":"LPS_Hz_7d_2",
"Factors":{"Treatment":"LPS + Hz","Time":"7d"}
},
{
"Subject ID":"-",
"Sample ID":"LPS_Hz_7d_3",
"Factors":{"Treatment":"LPS + Hz","Time":"7d"}
}
],
"COLLECTION":{"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_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"},

"SAMPLEPREP":{"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."},

"CHROMATOGRAPHY":{"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.","CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Shimadzu Nexera X2","COLUMN_NAME":"Ascentis Express C18 (100 x 4.6, 2.7)","METHODS_FILENAME":"PR_CH_Methods.pdf"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"INSTRUMENT_NAME":"ABI Sciex 5600 TripleTOF","INSTRUMENT_TYPE":"QTOF","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","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).","MS_RESULTS_FILE":"ST001255_AN002084_Results.txt UNITS:peak intensity Has m/z:Yes Has RT:Yes RT units:Seconds"}

}