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"PROJECT":{"PROJECT_TITLE":"Fish-oil supplementation in pregnancy, child metabolomics and asthma risk","PROJECT_TYPE":"Untargeted metabolomics","PROJECT_SUMMARY":"We investigated potential metabolic mechanisms using untargeted liquid chromatography-mass spectrometry-based metabolomics on 577 plasma samples collected at age 6 months in the offspring of mothers participating in the n-3 LCPUFA randomised controlled trial. First, associations between the n-3 LCPUFA supplementation groups and child metabolite levels were investigated using univariate regression models and data-driven partial least square discriminant analyses (PLS-DA). Second, we analysed the association between the n-3 LCPUFA metabolomic profile and asthma development using Cox-regression. Third, we conducted mediation analyses to investigate whether the protective effect of n-3 LCPUFA on asthma was mediated via the metabolome.","INSTITUTE":"Copenhagen Prospective Studies on Asthma in Childhood, Herlev and Gentofte Hospital, University of Copenhagen, Copenhagen, Denmark","DEPARTMENT":"Copenhagen Prospective Studies on Asthma in Childhood, Herlev and Gentofte Hospital","LAST_NAME":"Rago","FIRST_NAME":"Daniela","ADDRESS":"Ledreborg Alle 34, Gentofte, DK, 2820, Denmark","EMAIL":"daniela.rago@dbac.dk","PHONE":"+45-39777360"},

"STUDY":{"STUDY_TITLE":"Fish-oil supplementation in pregnancy, child metabolomics and asthma risk","STUDY_SUMMARY":"We investigated potential metabolic mechanisms using untargeted liquid chromatography-mass spectrometry-based metabolomics on 577 plasma samples collected at age 6 months in the offspring of mothers participating in the n-3 LCPUFA randomized controlled trial. First, associations between the n-3 LCPUFA supplementation groups and child metabolite levels were investigated using univariate regression models and data-driven partial least square discriminant analyses (PLS-DA). Second, we analyzed the association between the n-3 LCPUFA metabolomic profile and asthma development using Cox-regression. Third, we conducted mediation analyses to investigate whether the protective effect of n-3 LCPUFA on asthma was mediated via the metabolome","INSTITUTE":"Copenhagen Prospective Studies on Asthma in Childhood, Herlev and Gentofte Hospital, University of Copenhagen","DEPARTMENT":"Copenhagen Prospective Studies on Asthma in Childhood, Herlev and Gentofte Hospital","LAST_NAME":"Rago","FIRST_NAME":"Daniela","ADDRESS":"Ledreborg Alle 34, DK-2820 Gentofte; Denmark","EMAIL":"daniela.rago@dbac.dk","PHONE":"(+45) 39777360"},

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"COLLECTION":{"COLLECTION_SUMMARY":"The blood sample was collected in an EDTA tube and left at room temperature for 30 min and thereafter spun down for 10 min at 4000 rpm. The supernatant was collected and stored at -80 C until further analysis.","SAMPLE_TYPE":"Blood (plasma)","STORAGE_CONDITIONS":"-80℃"},

"TREATMENT":{"TREATMENT_SUMMARY":"The pregnant women were enrolled in a double-blind, randomized controlled trial (DB-RCT)3. At enrollment during pregnancy week 22-26, the women were randomly assigned in a 1:1 ratio to receive 2.4 g per day of n−3 long-chain polyunsaturated fatty acids (LCPUFAs) in triacylglycerol form from fish oil capsules containing 55% eicosapentaenoic acid (20:5n–3, EPA) and 37% docosahexaenoic acid (22:6n–3, DHA (Incromega TG33/22, Croda, Health Care) or placebo in the form of lookalike olive oil capsules, containing 72% n–9 oleic acid and 12% n−6 linoleic acid (Pharma-Tech A/S). The women continued taking the daily supplement until 1 week after delivery, where they visited the research unit with their child. A subgroup of the pregnant women (n=623) also participated in another DB-RCT with a nested 2x2 factorial design in which they were assigned to 2400 IU of vitamin D per day or placebo from pregnancy week 22-26 till 1 week postpartum on top of the recommended pregnancy supplement of 400 IU/d; i.e. the women received 2800 vs. 400 IU/d of vitamin D, which did not significantly affect the offspring’s risk of developing asthma"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"The sample preparation was done using the automated system MicroLab STAR® system from Hamilton Company. Prior to extraction, the samples were fortified with recovery standards for quality control (QC) purposes. Methanol was used to extract the metabolites and precipitate the proteins during vigorous shaking for 2 min (Glen Mills GenoGrinder 2000) followed by centrifugation. The resulting extract was divided into four aliquots to be subsequently analyzed by LC-MS/MS comprising of four platforms. Samples were placed on a TurboVap® (Zymark) to remove the organic solvent and stored overnight under nitrogen before preparation for LC-MS/MS analysis.","PROCESSING_STORAGE_CONDITIONS":"-80℃"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"HILIC","INSTRUMENT_NAME":"Waters Acquity","COLUMN_NAME":"Waters Acquity BEH Amide (150 x 2.1mm, 1.7um)","METHODS_FILENAME":"daniela_rago_CH_LC-MS_methods.pdf"},

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

"MS":{"INSTRUMENT_NAME":"Thermo Q Exactive Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","MS_COMMENTS":"The MS analysis alternated between MS and data-dependent MSn scans using dynamic exclusion. The scan range varied slighted between methods but covered 70-1000 m/z. Raw data was extracted, peak-identified and QC processed using Metabolon’s hardware and software. Compounds were identified by comparison to library entries of purified standards or recurrent unknown entities."},

"MS_METABOLITE_DATA":{
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