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FILES/RAW_FILES/2023_03_06_F069_16_6_NEG.raw.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15899/FILES/RAW_FILES/2023_03_10_F068_27_4_NEG.raw.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15899/FILES/RAW_FILES/2023_03_10_F023_18_1_NEG.raw.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15899/FILES/RAW_FILES/2023_03_04_F035_17_1_NEG.raw.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15899/FILES/RAW_FILES/2023_03_04_F046_13_3_NEG.raw.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15899/FILES/RAW_FILES/2023_03_10_F023_37_2_NEG.raw.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15899/FILES/RAW_FILES/2023_03_09_F064_12_4_NEG.raw.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15899/FILES/RAW_FILES/2023_03_09_BLANK_02_NEG.raw.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15899/FILES/RAW_FILES/2023_03_09_BLANK_03_NEG.raw.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15899/FILES/RAW_FILES/2023_03_09_BLANK_01_NEG.raw.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15899/FILES/RAW_FILES/2023_03_10_F071_15_2_NEG.raw.zipprimary200OKftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15899<p>Metabolite identification for set of reported features was originally performed in Pruski et al 2021. Target m/z features were putatively annotated using online databases including HMDB, Metlin, MMCD, KEGG and Lipidmaps with a 5 ppm tolerance for each compound. To account for potential imprecision in m/z measurement due to peak binning and data processing artefacts, raw m/z value were confirmed by inspection of the.raw data. Metabolite annotation was performed by searching the measured m/z ratios against METLIN (http://metlin.scripps.edu), Lipidmaps (http://www.lipidmaps.org) and the HMDB (http://www.hmdb.ca). Further structural elucidation was performed using MS/MS experiments via collision-induced dissociation on the LTQ-Discovery MS instrument (Thermo Scientific). For the annotation of metabolites, the MS/MS spectra were matched against spectral libraries from HMDB, NIST and METLIN that were compiled with either authentic standards or theoretical assignment. Identification of metabolites where MS/MS reference spectra were not available were annotated using chemical fragmentation rules. For the structural assignment of glycerophospholipid, fragments of the polar head group or the fatty acyl chains were investigated to confirm the annotation proposed by the databases and discriminate isomers. MSI levels of compound identification were further reported as suggested by the Metabolomics Standards Initiative (MSI).</p><p><br></p><p>References</p><p>Pruski, P., Correia, G.D.S., Lewis, H.V. <em>et al.</em> Direct on-swab metabolic profiling of vaginal microbiome host interactions during pregnancy and preterm birth. <em>Nat Commun</em><strong> 12</strong>, 5967 (2021). https://doi.org/10.1038/s41467-021-26215-w</p>MetaboLightsPublicDirect infusion MS - negative<p>Direct analysis of swabs using DESI-MS was performed using the methodology described in Pruski et al 2017 and Pruski et al 2021.</p><p><br></p><p>References</p><p>1 - Pruski, P., et al, Medical Swab Analysis Using Desorption Electrospray Ionization Mass Spectrometry: A Noninvasive Approach for Mucosal Diagnostics. Anal. Chem. (2017). https://doi.org/10.1021/acs.analchem.6b03405</p><p>2 - Pruski, P., Correia, G.D.S., Lewis, H.V. et al. Direct on-swab metabolic profiling of vaginal microbiome host interactions during pregnancy and preterm birth. Nat Commun (2021). https://doi.org/10.1038/s41467-021-26215-w</p>LACTIN-V (Lactobacillus crispatus CTV-05) leads to microbial and immune modulation in pregnant women at risk of spontaneous preterm birth: FLIP-1 study.GonÃÂçalo dos Santos CorreiaImperial College Londoncervicovaginal fluidmass spectrometry assay<p>Spectral profiles were acquired directly from swabs with an ambient Desorption electrospray ionisation mass spectrometry (DESI-MS) method. No extraction was performed. </p>Homo sapienshttps://www.ebi.ac.uk/metabolights/MTBLS15899David MacIntyre. Imperial College London. Institute of Reproductive and Developmental Biology, Hammersmith Hospital Campus, W12 0NN, London, UK. d.macintyre@imperial.ac.uk.Goncalo Correia. Imperial College London. Institute of Reproductive and Developmental Biology, Hammersmith Hospital Campus, W12 0NN, London, UK. gd2212@ic.ac.uk.<p>RAW files were converted to .mzML with proteoWizard, prior to processing using custom Python scripts. Individual scans were merged using a Python implementation of the MALDIquant 'binPeaks' algorithm and averaged to generate a single spectrum. The m/z values were re-calibrated using a linear function fitted robustly with the RANSAC algorithm and a reference list of previously annotated m/z values. The spectrum profiles were then assembled into a data matrix of intensities using again the MALDIquant binPeaks algorithm.</p>Batchgd2212@ic.ac.uk<p><strong>Study design&nbsp;</strong></p><p>Women at high risk for sPTB were recruited from preterm birth prevention clinics at Queen Charlotte’s and Chelsea Hospital between September 2019 and April 2021. Women provided written informed consent as part of the FLIP-1 study (Feasibility, safety, tolerance and assessment of LACTIN-V use In a cohort of Pregnant women at risk of preterm birth), which was ethically approved by the NHS Health Research Authority London-Surrey Borders Research Ethics Committee, REC 19/LO/1018. ClinicalTrials.gov ID, NCT03992534. Inclusion criteria were women at risk of preterm labour who were referred to the prematurity clinic, and exclusion criteria were HIV positive women, women unable to provide informed consent, women aged &lt;18, and women receiving antibiotic treatment within 1 week of recruitment.&nbsp;Women were eligible for recruitment between 11+0 and 15+6 weeks of pregnancy and were recruited at their first visit to the PTB prevention clinic, with a median gestational age of 13+5 (IQR 13+0-14+4).&nbsp;&nbsp;</p><p><br></p><p><strong>Study procedure and sample collection&nbsp;</strong></p><p>During the consent process, women were informed of the administration schedule for LACTIN-V use. The dosing schedule commenced with a loading dose phase consisting of five consecutive daily doses of LACTIN-V, followed by a maintenance phase consisting of six weekly doses of LACTIN-V. A supply of pre-filled applicators containing Lactobacillus crispatus CTV-05 powder at a dose of 2x 109 CFU preserved with inactive ingredients was provided to the participant. Consenting women were provided with 11 vaginal applicators to be stored at 2-8°C in a standard refrigerator. Each participant was asked to report adverse events and antibiotic use during pregnancy. Participants were followed up in the PTB prevention clinic, and vaginal swabs were collected prior to the loading dose (Timepoint 0, T0), and at 6 subsequent longitudinal timepoints; T1 (Median GA 15+1 [IQR 14+3-16+1]), T2 (18+1 [17+1-18+6]), T3 (20+2 [19+2-20+6]), T4 (28+2 [27+5-29+4]), T5 (36+1 [34+6-36+3]), and T6 (at delivery (38+3 [37+4-39+2]. Cervicovaginal fluid (CVF) was sampled using BBLTM CultureSwabTM MaxV Liquid Amies (Becton, Dickinson and Company, Oxford UK, for 16S metataxonomics) and Transwab (MW170 with rayon blue lid, Medical Wire &amp; Equipment, Corsham, UK, for DESI-MS metabolomics). Swabs were stored at -80°C until use. Standard clinical care was continued in the PTB prevention clinic, with the use of progesterone and/or cervical cerclage offered where clinically indicated. LACTIN-V was provided by OSEL Inc. California.&nbsp;</p><p><br></p>Metabolomicscervicovaginal fluidMetabolomicsuntargeted analysisPreterm Birthquality control materialpregnancyHomo sapienssample preparation blankmicrobiomeWaters ACQUITY RDa Detectorexperimental sampleuntargeted metabolite profilingdesorption electrospray ionisation mass spectrometryVaginalcervicovaginal fluidMetabolomicsuntargeted analysisPreterm Birthquality control materialpregnancyHomo sapienssample preparation blankmicrobiomeWaters ACQUITY RDa Detectorexperimental sampleuntargeted metabolite profilingdesorption electrospray ionisation mass spectrometryVaginal<p>DESI-MS analyses for the FLIP-1 cohort were performed using an ACQUITY RDa/BioAccord Time-of-Flight mass spectrometer (Waters Corporation, Wilmslow, UK).&nbsp;For swab analysis and as part of the DESI stage, a roboticised sampling device was used. Swabs were placed into a rotating holder positioned orthogonally in front of the MS inlet capillary with a swab–capillary distance of approximately 2 mm. The DESI sprayer tip was pointed to the swab centre with a tip-sample distance of 1.5–2 mm and a distance between the tip and the inlet capillary of 2 mm. The entire surface of the swabs was analysed by DESI-MS through clockwise rotation of the swab toward the MS capillary using a methanol/water (95:5, v/v) solvent at 30 µL/min. Data were acquired in both positive and negative ion modes in the m/z range of 50-2000. Data acquisition parameters were: capillary voltage 0.8-1.2 kV, gas pressure at 4 bar, source temperature at 120°C, and gas desolvation at 450°C. Samples from individual patients were analysed consecutively, but the order in which different patients' samples were analysed was randomised by gestational age, parity, and microbial composition.</p>Glutamyl-LeucinePG(18:1/18:2)Leucyl-ThreonineValyl-GlutamineValyl-PhenylalanineLyso-PE(18:2)Leucyl-GlutamineLeucyl-SerineLignoceric acid (C24:0)N2-Acetyl-ornithineOleic Acid (C18:1)N-palmitoyl-cysteineN-palmitoyl glutamineOctadecyl-oxy-octadecanoic acid (SAHSA)LactoylcysteineOctadecanoyloxamideOxoprolineGlutamateStearic acid (18:0)N-palmitoyl methionineLyso-PE(18:1)PI(20:4/18:0)Leucyl-glycyl-glycineCerotic acid (C26:0)Leucyl-AlanineGalbeta-Cer(d40:1)OxypurinolDG(36:3)Lyso-PE(20:4)Valyl-SerineLyso-PS(18:0)Dihydroxy-octadecadienoic acidLeucineNervonic acid (C24:1)Linolenic Acid (C18:3)Aspartic acidNonadecanoic acid (C19:0)Phosphatidyl glycerolPS(18:0/18:1)PhenylalanineN-AcetylputrescinePhenyl sulfoxidePS(16:0/18:1)HODEValyl-AlanineLinoleic acid (C18:2)Oxo-octadecadienoic acidAsparagineGlycyl-PhenylalanineGlutaminefalseLACTIN-V (Lactobacillus crispatus CTV-05) leads to microbial and immune modulation in pregnant women at risk of spontaneous preterm birth: FLIP-1 study<p>Preterm birth (PTB) remains a leading cause of neonatal morbidity and mortality, with no new preventive therapeutics introduced since the 1970s. Evidence increasingly implicates adverse vaginal microbiome-immune interactions in its aetiology. We report the effects of the live biotherapeutic, Lactobacillus crispatus CTV-05 (LACTIN-V), in an observational study of pregnant individuals at high-risk of PTB (n=61). LACTIN-V treatment was associated with a shift towards L. crispatus-dominated microbiota, which was sustained at eight weeks post-cessation and confirmed to be viable CTV-05. Engraftment of LACTIN-V was accompanied by significant reductions in cervicovaginal fluid concentrations of IL-1β, IL-6, C3a, C3b/iC3b, C4, and Factor H, changes not seen in untreated time-matched controls. We also highlight the utility of metabolic profiling by Desorption Electrospray Ionisation in identifying individuals most likely to benefit from treatment and for assessing therapeutic response. The reduced PTB rate observed with LACTIN-V administration supports its use in preventing microbiome–immune–mediated PTB.</p>2026-10-022026-10-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:22653CHEBI:22660CHEBI:31009CHEBI:85700CHEBI:176392CHEBI:178203CHEBI:18237CHEBI:28300CHEBI:73506CHEBI:73912CHEBI:36328CHEBI:178206CHEBI:25017CHEBI:73527CHEBI:157865CHEBI:74538CHEBI:73523CHEBI:178207CHEBI:28866CHEBI:17351CHEBI:25048CHEBI:64575CHEBI:76233CHEBI:64569CHEBI:72408CHEBI:17768CHEBI:165552CHEBI:42339CHEBI:21780CHEBI:16543CHEBI:44247CHEBI:39246CHEBI:178209CHEBI:178210CHEBI:16196CHEBI:178211CHEBI:25801CHEBI:28315CHEBI:89348CHEBI:177532CHEBI:28044CHEBI:15978CHEBI:89110CHEBI:34086CHEBI:79096CHEBI:28842CHEBI:75008CHEBI:75010CHEBI:75016CHEBI:177274