{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001655","ANALYSIS_ID":"AN002702","VERSION":"1","CREATED_ON":"January 20, 2021, 9:51 am"},

"PROJECT":{"PROJECT_TITLE":"Characterization of anaphylaxis reveals different metabolic changes depending on severity and triggers.","PROJECT_SUMMARY":"Despite its increasing incidence, the underlying molecular processes of anaphylaxis remain unclear and there are not known biomarkers for appropriate diagnosis. The mechanism associated to the reactions still needs to be clarified in humans. The rapid onset and potentially fatal outcome in the absence of managed treatment, prevent its study and prompt obvious technical and ethical implications. Twenty episodes of anaphylaxis were analyzed. Sera was collected at different times: during the acute phase (T1), the recovery phase (T2) and around 2-3 months after the anaphylactic reaction (T0). The analysis included untargeted metabolomics combining liquid chromatography coupled to mass spectrometry (LC-MS) and proton-nuclear magnetic resonance (1H-NMR). Reactions were classified according to the trigger (food and/or drug) and severity (moderate and severe). “Food T1 vs T2” and “moderate T1 vs T2” anaphylaxis comparisons showed clear metabolic patterns during the onset of an anaphylactic reaction, which differed from those induced by drugs, food+drug or severe anaphylaxis “T1 vs T2”. Moreover, the model of food anaphylaxis was able to distinguish the well-characterized IgE (beta-lactam) from non-IgE- mediated anaphylaxis (NSAIDs), suggesting a differential metabolic pathway associated with the mechanism of action. Moreover, metabolic differences between “moderate vs severe” at T1 and T0 were studied. Among the metabolites, glucose, lipids, cortisol, betaine and oleamide were observed altered. The results of the study provide the first evidence that different anaphylactic triggers, induce differential metabolic changes. Besides, the basal status might identify high risk patients, thus opening new ways to understand, diagnose and treat anaphylaxis.","INSTITUTE":"Hospital Universitari i Politècnic La Fe","DEPARTMENT":"Subdirección Médica, Departament de Salut València La Fe.","LAST_NAME":"Hernández Fernández de Rojas","FIRST_NAME":"Dolores","ADDRESS":"Avinguda de Fernando Abril Martorell, 106, 46026 València, Valencia, España.","EMAIL":"hernandez_dol@gva.es","PHONE":"91 372 47 00 ext. 4662"},

"STUDY":{"STUDY_TITLE":"Characterization of anaphylaxis reveals different metabolic changes depending on severity and triggers.","STUDY_SUMMARY":"Background: Despite its increasing incidence, the underlying molecular processes of anaphylaxis remain unclear and there are not known biomarkers for appropriate diagnosis. The mechanism associated to the reactions still needs to be clarified in humans. The rapid onset and potentially fatal outcome in the absence of managed treatment, prevent its study and prompt obvious technical and ethical implications. Methods: Twenty episodes of anaphylaxis were analyzed. Sera was collected at different times: during the acute phase (T1), the recovery phase (T2) and around 2-3 months after the anaphylactic reaction (T0). The analysis included untargeted metabolomics combining liquid chromatography coupled to mass spectrometry (LC-MS) and proton-nuclear magnetic resonance (1H-NMR). Reactions were classified according to the trigger (food and/or drug) and severity (moderate and severe). Results: “Food T1 vs T2” and “moderate T1 vs T2” anaphylaxis comparisons showed clear metabolic patterns during the onset of an anaphylactic reaction, which differed from those induced by drugs, food+drug or severe anaphylaxis “T1 vs T2”. Moreover, the model of food anaphylaxis was able to distinguish the well-characterized IgE (beta-lactam) from non-IgE- mediated anaphylaxis (NSAIDs), suggesting a differential metabolic pathway associated with the mechanism of action. Moreover, metabolic differences between “moderate vs severe” at T1 and T0 were studied. Among the metabolites, glucose, lipids, cortisol, betaine and oleamide were observed altered. Conclusions: The results of the study provide the first evidence that different anaphylactic triggers, induce differential metabolic changes. Besides, the basal status might identify high risk patients, thus opening new ways to understand, diagnose and treat anaphylaxis.","INSTITUTE":"The Centre of Metabolomics and Bioanalysis","DEPARTMENT":"Analytical chemistry","LAST_NAME":"Obeso Montero","FIRST_NAME":"David","ADDRESS":"Av. de Montepríncipe, s/n","EMAIL":"david.obesomontero@beca.ceu.es","PHONE":"607535650","NUM_GROUPS":"2 groups","TOTAL_SUBJECTS":"20"},

"SUBJECT":{"SUBJECT_TYPE":"Human","SUBJECT_SPECIES":"Homo sapiens","TAXONOMY_ID":"9606","GENDER":"Male and female"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"P1_t0",
"Factors":{"Trigger":"Drug","Severity":"Moderate","Time":"Time 0"},
"Additional sample data":{"RAW_FILE_NAME":"P1_t0.d"}
},
{
"Subject ID":"-",
"Sample ID":"P1_t1",
"Factors":{"Trigger":"Drug","Severity":"Moderate","Time":"Time 1"},
"Additional sample data":{"RAW_FILE_NAME":"P1_t1.d"}
},
{
"Subject ID":"-",
"Sample ID":"P1_t2",
"Factors":{"Trigger":"Drug","Severity":"Moderate","Time":"Time 2"},
"Additional sample data":{"RAW_FILE_NAME":"P1_t2.d"}
},
{
"Subject ID":"-",
"Sample ID":"P10_t1",
"Factors":{"Trigger":"Food","Severity":"Moderate","Time":"Time 1"},
"Additional sample data":{"RAW_FILE_NAME":"P10_t1.d"}
},
{
"Subject ID":"-",
"Sample ID":"P10_t2",
"Factors":{"Trigger":"Food","Severity":"Moderate","Time":"Time 2"},
"Additional sample data":{"RAW_FILE_NAME":"P10_t2.d"}
},
{
"Subject ID":"-",
"Sample ID":"P11_t0",
"Factors":{"Trigger":"Food","Severity":"Severe","Time":"Time 0"},
"Additional sample data":{"RAW_FILE_NAME":"P11_t0.d"}
},
{
"Subject ID":"-",
"Sample ID":"P11_t1",
"Factors":{"Trigger":"Food","Severity":"Severe","Time":"Time 1"},
"Additional sample data":{"RAW_FILE_NAME":"P11_t1.d"}
},
{
"Subject ID":"-",
"Sample ID":"P11_t2",
"Factors":{"Trigger":"Food","Severity":"Severe","Time":"Time 2"},
"Additional sample data":{"RAW_FILE_NAME":"P11_t2.d"}
},
{
"Subject ID":"-",
"Sample ID":"P12_t0",
"Factors":{"Trigger":"Idiopatic","Severity":"Severe","Time":"Time 0"},
"Additional sample data":{"RAW_FILE_NAME":"P12_t0.d"}
},
{
"Subject ID":"-",
"Sample ID":"P12_t1",
"Factors":{"Trigger":"Idiopatic","Severity":"Severe","Time":"Time 1"},
"Additional sample data":{"RAW_FILE_NAME":"P12_t1.d"}
},
{
"Subject ID":"-",
"Sample ID":"P12_t2",
"Factors":{"Trigger":"Idiopatic","Severity":"Severe","Time":"Time 2"},
"Additional sample data":{"RAW_FILE_NAME":"P12_t2.d"}
},
{
"Subject ID":"-",
"Sample ID":"P13_t0",
"Factors":{"Trigger":"Drug","Severity":"Severe","Time":"Time 0"},
"Additional sample data":{"RAW_FILE_NAME":"P13_t0.d"}
},
{
"Subject ID":"-",
"Sample ID":"P13_t1",
"Factors":{"Trigger":"Drug","Severity":"Severe","Time":"Time 1"},
"Additional sample data":{"RAW_FILE_NAME":"P13_t1.d"}
},
{
"Subject ID":"-",
"Sample ID":"P13_t2",
"Factors":{"Trigger":"Drug","Severity":"Severe","Time":"Time 2"},
"Additional sample data":{"RAW_FILE_NAME":"P13_t2.d"}
},
{
"Subject ID":"-",
"Sample ID":"P14_t0",
"Factors":{"Trigger":"Drug","Severity":"Severe","Time":"Time 0"},
"Additional sample data":{"RAW_FILE_NAME":"P14_t0.d"}
},
{
"Subject ID":"-",
"Sample ID":"P14_t1",
"Factors":{"Trigger":"Drug","Severity":"Severe","Time":"Time 1"},
"Additional sample data":{"RAW_FILE_NAME":"P14_t1.d"}
},
{
"Subject ID":"-",
"Sample ID":"P14_t2",
"Factors":{"Trigger":"Drug","Severity":"Severe","Time":"Time 2"},
"Additional sample data":{"RAW_FILE_NAME":"P14_t2.d"}
},
{
"Subject ID":"-",
"Sample ID":"P15_t0",
"Factors":{"Trigger":"Drug","Severity":"Mild","Time":"Time 0"},
"Additional sample data":{"RAW_FILE_NAME":"P15_t0.d"}
},
{
"Subject ID":"-",
"Sample ID":"P15_t1",
"Factors":{"Trigger":"Drug","Severity":"Mild","Time":"Time 1"},
"Additional sample data":{"RAW_FILE_NAME":"P15_t1.d"}
},
{
"Subject ID":"-",
"Sample ID":"P16_t0",
"Factors":{"Trigger":"Food","Severity":"Moderate","Time":"Time 0"},
"Additional sample data":{"RAW_FILE_NAME":"P16_t0.d"}
},
{
"Subject ID":"-",
"Sample ID":"P16_t1",
"Factors":{"Trigger":"Food","Severity":"Moderate","Time":"Time 1"},
"Additional sample data":{"RAW_FILE_NAME":"P16_t1.d"}
},
{
"Subject ID":"-",
"Sample ID":"P16_t2",
"Factors":{"Trigger":"Food","Severity":"Moderate","Time":"Time 2"},
"Additional sample data":{"RAW_FILE_NAME":"P16_t2.d"}
},
{
"Subject ID":"-",
"Sample ID":"P17_t0",
"Factors":{"Trigger":"Drug","Severity":"Moderate","Time":"Time 0"},
"Additional sample data":{"RAW_FILE_NAME":"P17_t0.d"}
},
{
"Subject ID":"-",
"Sample ID":"P17_t1",
"Factors":{"Trigger":"Drug","Severity":"Moderate","Time":"Time 1"},
"Additional sample data":{"RAW_FILE_NAME":"P17_t1.d"}
},
{
"Subject ID":"-",
"Sample ID":"P17_t2",
"Factors":{"Trigger":"Drug","Severity":"Moderate","Time":"Time 2"},
"Additional sample data":{"RAW_FILE_NAME":"P17_t2.d"}
},
{
"Subject ID":"-",
"Sample ID":"P18_t0",
"Factors":{"Trigger":"Food","Severity":"Moderate","Time":"Time 0"},
"Additional sample data":{"RAW_FILE_NAME":"P18_t0.d"}
},
{
"Subject ID":"-",
"Sample ID":"P18_t1",
"Factors":{"Trigger":"Food","Severity":"Moderate","Time":"Time 1"},
"Additional sample data":{"RAW_FILE_NAME":"P18_t1.d"}
},
{
"Subject ID":"-",
"Sample ID":"P18_t2",
"Factors":{"Trigger":"Food","Severity":"Moderate","Time":"Time 2"},
"Additional sample data":{"RAW_FILE_NAME":"P18_t2.d"}
},
{
"Subject ID":"-",
"Sample ID":"P19_t0",
"Factors":{"Trigger":"Drug","Severity":"Severe","Time":"Time 0"},
"Additional sample data":{"RAW_FILE_NAME":"P19_t0.d"}
},
{
"Subject ID":"-",
"Sample ID":"P19_t1",
"Factors":{"Trigger":"Drug","Severity":"Severe","Time":"Time 1"},
"Additional sample data":{"RAW_FILE_NAME":"P19_t1.d"}
},
{
"Subject ID":"-",
"Sample ID":"P19_t2",
"Factors":{"Trigger":"Drug","Severity":"Severe","Time":"Time 2"},
"Additional sample data":{"RAW_FILE_NAME":"P19_t2.d"}
},
{
"Subject ID":"-",
"Sample ID":"P2_t0",
"Factors":{"Trigger":"Food","Severity":"Moderate","Time":"Time 0"},
"Additional sample data":{"RAW_FILE_NAME":"P2_t0.d"}
},
{
"Subject ID":"-",
"Sample ID":"P2_t1",
"Factors":{"Trigger":"Food","Severity":"Moderate","Time":"Time 1"},
"Additional sample data":{"RAW_FILE_NAME":"P2_t1.d"}
},
{
"Subject ID":"-",
"Sample ID":"P2_t2",
"Factors":{"Trigger":"Food","Severity":"Moderate","Time":"Time 2"},
"Additional sample data":{"RAW_FILE_NAME":"P2_t2.d"}
},
{
"Subject ID":"-",
"Sample ID":"P20_t0",
"Factors":{"Trigger":"Other","Severity":"Moderate","Time":"Time 0"},
"Additional sample data":{"RAW_FILE_NAME":"P20_t0.d"}
},
{
"Subject ID":"-",
"Sample ID":"P20_t1",
"Factors":{"Trigger":"Other","Severity":"Moderate","Time":"Time 1"},
"Additional sample data":{"RAW_FILE_NAME":"P20_t1.d"}
},
{
"Subject ID":"-",
"Sample ID":"P3_t0",
"Factors":{"Trigger":"Drug","Severity":"Moderate","Time":"Time 0"},
"Additional sample data":{"RAW_FILE_NAME":"P3_t0.d"}
},
{
"Subject ID":"-",
"Sample ID":"P3_t1",
"Factors":{"Trigger":"Drug","Severity":"Moderate","Time":"Time 1"},
"Additional sample data":{"RAW_FILE_NAME":"P3_t1.d"}
},
{
"Subject ID":"-",
"Sample ID":"P4_t0",
"Factors":{"Trigger":"Drug","Severity":"Severe","Time":"Time 0"},
"Additional sample data":{"RAW_FILE_NAME":"P4_t0.d"}
},
{
"Subject ID":"-",
"Sample ID":"P4_t1",
"Factors":{"Trigger":"Drug","Severity":"Severe","Time":"Time 1"},
"Additional sample data":{"RAW_FILE_NAME":"P4_t1.d"}
},
{
"Subject ID":"-",
"Sample ID":"P4_t2",
"Factors":{"Trigger":"Drug","Severity":"Severe","Time":"Time 2"},
"Additional sample data":{"RAW_FILE_NAME":"P4_t2.d"}
},
{
"Subject ID":"-",
"Sample ID":"P5_t1",
"Factors":{"Trigger":"Idiopatic","Severity":"Moderate","Time":"Time 1"},
"Additional sample data":{"RAW_FILE_NAME":"P5_t1.d"}
},
{
"Subject ID":"-",
"Sample ID":"P5_t2",
"Factors":{"Trigger":"Idiopatic","Severity":"Moderate","Time":"Time 2"},
"Additional sample data":{"RAW_FILE_NAME":"P5_t2.d"}
},
{
"Subject ID":"-",
"Sample ID":"P6_t0",
"Factors":{"Trigger":"Idiopatic","Severity":"Severe","Time":"Time 0"},
"Additional sample data":{"RAW_FILE_NAME":"P6_t0.d"}
},
{
"Subject ID":"-",
"Sample ID":"P6_t1",
"Factors":{"Trigger":"Idiopatic","Severity":"Severe","Time":"Time 1"},
"Additional sample data":{"RAW_FILE_NAME":"P6_t1.d"}
},
{
"Subject ID":"-",
"Sample ID":"P6_t2",
"Factors":{"Trigger":"Idiopatic","Severity":"Severe","Time":"Time 2"},
"Additional sample data":{"RAW_FILE_NAME":"P6_t2.d"}
},
{
"Subject ID":"-",
"Sample ID":"P7_t0",
"Factors":{"Trigger":"Drug","Severity":"Severe","Time":"Time 0"},
"Additional sample data":{"RAW_FILE_NAME":"P7_t0.d"}
},
{
"Subject ID":"-",
"Sample ID":"P7_t1",
"Factors":{"Trigger":"Drug","Severity":"Severe","Time":"Time 1"},
"Additional sample data":{"RAW_FILE_NAME":"P7_t1.d"}
},
{
"Subject ID":"-",
"Sample ID":"P7_t2",
"Factors":{"Trigger":"Drug","Severity":"Severe","Time":"Time 2"},
"Additional sample data":{"RAW_FILE_NAME":"P7_t2.d"}
},
{
"Subject ID":"-",
"Sample ID":"P8_t0",
"Factors":{"Trigger":"Other","Severity":"Moderate","Time":"Time 0"},
"Additional sample data":{"RAW_FILE_NAME":"P8_t0.d"}
},
{
"Subject ID":"-",
"Sample ID":"P8_t1",
"Factors":{"Trigger":"Other","Severity":"Moderate","Time":"Time 1"},
"Additional sample data":{"RAW_FILE_NAME":"P8_t1.d"}
},
{
"Subject ID":"-",
"Sample ID":"P8_t2",
"Factors":{"Trigger":"Other","Severity":"Moderate","Time":"Time 2"},
"Additional sample data":{"RAW_FILE_NAME":"P8_t2.d"}
},
{
"Subject ID":"-",
"Sample ID":"P9_t0",
"Factors":{"Trigger":"Food","Severity":"Moderate","Time":"Time 0"},
"Additional sample data":{"RAW_FILE_NAME":"P9_t0.d"}
},
{
"Subject ID":"-",
"Sample ID":"P9_t1",
"Factors":{"Trigger":"Food","Severity":"Moderate","Time":"Time 1"},
"Additional sample data":{"RAW_FILE_NAME":"P9_t1.d"}
},
{
"Subject ID":"-",
"Sample ID":"P9_t2",
"Factors":{"Trigger":"Food","Severity":"Moderate","Time":"Time 2"},
"Additional sample data":{"RAW_FILE_NAME":"P9_t2.d"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"A prospective clinical and observational study of patients with anaphylactic reactions was performed. Patients of all ages and both sexes were recruited at outpatient clinics and the departments of Emergency, and other services at Hospital La Fe. All fulfilled clinical criteria of anaphylaxis and severity was graded following the classification by Brown, et al3. Patients were classified as food, drug or idiopathic origin, as well as in mild, moderate or severe according to the number of organs affected and clinical symptoms. The allergy evaluation was conducted by the Allergology Service of Hospital La Fe. The ethical committee approved the study protocol and all subjects were informed and provided written consent. Serum samples were taken during the acute moment of the reaction at the first moment of medical attention (< 2h, hereafter referred as ‘T1’), and after clinical recovery (approximately 2-4h later, referred to as ‘T2’). Patients were treated according to the Galaxy 2016 practical guide, using all necessary drugs to rescue them. Samples were collected, following specific standard operating procedures (SOPs)31-33. Briefly, these were collected in a vacutainer tube (Ref. 368965) and processed within the first 30 to 60 min after blood extraction. Sample aliquots were stored at -80ºC until further analyses. Subsequently, between 2-3 months after the anaphylaxis, a serum sample was taken when the allergy evaluation was performed (basal state, called ‘T0’).","SAMPLE_TYPE":"Blood (serum)","STORAGE_CONDITIONS":"-80℃"},

"TREATMENT":{"TREATMENT_SUMMARY":"Patients were treated according to the Galaxy 2016 practical guide, using all necessary drugs to rescue them."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"After thawing the samples, 150 µL of cold acetonitrile (0.1% formic acid, v/v) was added to 50 µL of serum, vortex and kept at -20ºC for 30 min for protein precipitation. Then, 25 µl of cleaned supernatant were transferred to a 96 well-plate for UPLC-MS analysis. Finally, 125 µL of H2O (0.1% formic acid, v/v), and 2 µL of internal standard (IS) mix solution, containing reserpine, leucine enkephaline, caffeine-d9 and phenylalanine-d5 in H20:CH3OH (1:1, 0.1% v/v formic acid) at 20 µM were added to each sample. Blank samples were prepared by replacing serum by ultrapure H20 in order to identify potential artefacts from the tube, reagents and other materials. A quality control (QC) was prepared by mixing 10 µL from each prepared sample. Samples were randomly injected in the chromatographic system (UPLC-ToF-MS) injection a QC every 6 serum samples in order to avoid intra-batch variability, as well as to enhance quality and reproducibility. Blank analysis was performed at the beginning and at end of the sequence. Sample stability and analytical drift were investigated through the internal standard intensities."},

"CHROMATOGRAPHY":[],

"ANALYSIS":{"ANALYSIS_TYPE":"NMR"},

"NM":{"INSTRUMENT_NAME":"Bruker 500MHz spectrometer","INSTRUMENT_TYPE":"FT-NMR","NMR_EXPERIMENT_TYPE":"1D-1H","SPECTROMETER_FREQUENCY":"500MHz","NMR_RESULTS_FILE":"ST001655_AN002702_Results.txt UNITS:ppm"}

}