Comprehensive lipid profiling of early stage oocytes and embryos by MRM profiling

Lima, Camila Bruna de; Ferreira, Christina R.; Milazzotto, Marcella Pécora; Sobreira, Tiago J. P.; Vireque, A.A.; Cooks, R. Graham

doi:10.1002/jms.4301

Cited by 52 publications

(46 citation statements)

References 26 publications

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“…It has previously been tested on several model systems, including Parkinson's disease, 36 polycystic ovarian syndrome, 37 diet compliance, 38 colostrum uptake, 39 atopic dermatitis, 40 and embryo metabolism. 41 The results of MRM-Profiling have proven to be broadly consistent with those from pathological or clinical analysis.…”

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confidence: 56%

Multiple Reaction Monitoring Profiling (MRM-Profiling) of Lipids To Distinguish Strain-Level Differences in Microbial Resistance in Escherichia coli

et al. 2019

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The worldwide increase in antimicrobial resistance is due to antibiotic over-use in agriculture and over-prescription in medicine. For appropriate and timely patient support, faster diagnosis of antimicrobial resistance is required. Current methods for bacterial identification rely on genomics and proteomics and use comparisons with databases of known strains, but the diagnostic value of metabolites and lipids has not been explored significantly. Standard mass spectrometry/chromatography methods involve multiple dilutions during sample preparation and separation. To increase the amount of chemical information acquired and the speed of analysis of lipids, multiple reaction monitoring profiling (MRM-Profiling) has been applied. The MRM-Profiling workflow includes a discovery stage and a screening stage. The discovery stage employs precursor ion (PREC) and neutral loss (NL) scans to screen representative pooled samples for functional groups associated with particular lipid classes. The information from the first stage is organized in precursor/product ion pairs, or MRMs, and the screening stage rapidly interrogates individual samples for these MRMs. In this study, we performed MRM-Profiling of lipid extracts from four different strains of Escherichia coli cultured with amoxicillin or amoxicillin/clavulanate, a beta-lactam and beta-lactamase inhibitor, respectively. T-tests, analysis of variance (ANOVA) and receiver operating characteristic (ROC) curves were used to determine the significance of each MRM. Principal component analysis (PCA) was applied to distinguish different strains cultured under conditions that allowed or disallowed development of bacterial resistance. The results demonstrate that MRM-Profiling distinguishes the lipid profiles of resistant and non-resistant E. coli strains. Annually there are over 1.2 billion health care visits in the United States of which 12.6% result in ~154 million antibiotic prescriptions for bacterial infections. 1 Urinary, respiratory and skin infections are most common. 2 The top pathogens responsible for 80 to 90% of infections are Escherichia coli, Staphylococcus aureus, and Klebsiella pneumoniae, the latter two of which are part of the ESKAPE group of pathogens which also include Enterococcus faecium, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species. 3-5 Infections are especially common in certain care settings and patient populations where they cause high recurrence rates and great severity of infections. 6-8 Potential infections are frequently treated prior to confirmation of the presence of pathogens, leading to over-prescription which is strongly associated with the development of antibioticresistance and may eradicate potentially commensal microbiome populations, resulting in dysfunction against future pathogens. 9-12 High rates of infection recurrence and the emergence of pathogens with antimicrobial resistance are found among patients with frequent antibiotic dosing. 11,13,14 There therefore remains a need for rapid bacterial identificati...

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supporting

confidence: 56%

Multiple Reaction Monitoring Profiling (MRM-Profiling) of Lipids To Distinguish Strain-Level Differences in Microbial Resistance in Escherichia coli

et al. 2019

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“…High-throughput lipid profiling of tissue, feces, plasma, and urine was performed by direct injection mass spectrometry (DIMS) as previously described ( Pereyra et al, 2017 ). Briefly, lipids were extracted using Bligh and Dyer Method ( Bligh and Dyer, 1959 ) and both, the lipid and the polar phases were dried separately, resuspended in ACN:MeOH:NH4Ac and directly injected via a micro-autosampler (G1377A) into a triple quadrupole mass spectrometer (QQQ6410 from Agilent Technologies, San Jose, CA) operated in the positive ion mode and equipped with ESI ion source Data was collected in multiple reaction monitoring mode ( de Lima et al, 2018 ; Franco et al, 2018 ; Pereyra et al, 2017 ). Ion intensities acquired by an in-house script were further normalized to sample protein concentration for tissues or extraction volume for biological fluids.…”

Section: Methodsmentioning

confidence: 99%

Loss of Muscle Carnitine Palmitoyltransferase 2 Prevents Diet-Induced Obesity and Insulin Resistance despite Long-Chain Acylcarnitine Accumulation

et al. 2020

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“…Lipids were extracted from individual blastocysts (n = 90; minimal of 5 replicates per group) collected at 168hpi using the Bligh & Dyer method and subsequently analyzed by Multiple Reactions Monitoring Mass Spectrometry (MRM-Profiling) following the protocol described in 22 . We selected 3 classes of lipids to include in the present analysis: Free Fatty Acids (30 transitions, including C16:0 and C18:1), Triacylglycerols (99 transitions) and Cholesteryl esters (49 transitions).…”

Section: Relative Quantification Of Lipids In Bovine Blastocystsmentioning

confidence: 99%

The dynamics between in vitro culture and metabolism: embryonic adaptation to environmental changes

Lima

Santos

Ispada

et al. 2020

Sci Rep

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Previous studies have discussed the importance of an optimal range of metabolic activity during preimplantation development. To avoid factors than can trigger an undesirable trajectory, it is important to learn how nutrients and metabolites interact to help launching the correct developmental program of the embryo, and how much the in vitro culture system can impair this process. Here, using the bovine model, we describe a factorial experimental design used to investigate the biochemical and molecular signature of embryos in response to different combinations of morphological features—i.e. speed of development—and external stimuli during in vitro culture—i.e. different oxygen tensions and glucose supplementation. Our analyses demonstrate that the embryos present heterogeneous metabolic responses depending on early morphological phenotypes and the composition of their surroundings. However, despite the contribution of each single stimulus for the embryo phenotype, oxygen tension is determinant for such differences. The lower oxygen environment boosts the metabolism of embryos with faster kinetics, in particular those cultured in lower glucose concentrations.

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Comprehensive lipid profiling of early stage oocytes and embryos by MRM profiling

Cited by 52 publications

References 26 publications

Multiple Reaction Monitoring Profiling (MRM-Profiling) of Lipids To Distinguish Strain-Level Differences in Microbial Resistance in Escherichia coli

Multiple Reaction Monitoring Profiling (MRM-Profiling) of Lipids To Distinguish Strain-Level Differences in Microbial Resistance in Escherichia coli

Loss of Muscle Carnitine Palmitoyltransferase 2 Prevents Diet-Induced Obesity and Insulin Resistance despite Long-Chain Acylcarnitine Accumulation

The dynamics between in vitro culture and metabolism: embryonic adaptation to environmental changes

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