Isotopic tracing of glucose metabolites in human monocytes to assess changes in inflammatory conditions.

STAR Protoc

Institute of Pharmacy, Freie Universität Berlin, Königin-Luise-Str. 2+4, 14195 Berlin, Germany. Electronic address:

Published: December 2022

Differences in metabolic profiles can link to functional changes of immune cells in disease conditions. Here, we detail a protocol for the detection and quantitation of 19 metabolites in one analytical run. We provide the parameters for chromatographic separation and mass spectrometric analysis of isotopically labeled and unlabeled metabolites. We include steps for incubation and sample preparation of PBMCs and monocytes. This protocol overcomes the chromatographic challenges caused by the chelating properties of some metabolites.

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9519621PMC
http://dx.doi.org/10.1016/j.xpro.2022.101715DOI Listing

Publication Analysis

Top Keywords

isotopic tracing
4
tracing glucose
4
metabolites
4
glucose metabolites
4
metabolites human
4
human monocytes
4
monocytes assess
4
assess changes
4
changes inflammatory
4
inflammatory conditions
4

Similar Publications

The Epstein-Barr virus (EBV) infects nearly 90% of adults globally and is linked to over 200,000 annual cancer cases. Immunocompromised individuals from conditions such as primary immune disorders, HIV, or posttransplant immunosuppressive therapies are particularly vulnerable because of EBV's transformative capability. EBV remodels B cell metabolism to support energy, biosynthetic precursors, and redox equivalents necessary for transformation.

View Article and Find Full Text PDF

Ketogenesis is a dynamic metabolic conduit supporting hepatic fat oxidation particularly when carbohydrates are in short supply. Ketone bodies may be recycled into anabolic substrates, but a physiological role for this process has not been identified. Here, we use mass spectrometry-based C-isotope tracing and shotgun lipidomics to establish a link between hepatic ketogenesis and lipid anabolism.

View Article and Find Full Text PDF

Stable-isotope resolved metabolomics (SIRM) is a powerful approach for characterizing metabolic states in cells and organisms. By incorporating isotopes, such as C, into substrates, researchers can trace reaction rates across specific metabolic pathways. Integrating metabolomics data with gene expression profiles further enriches the analysis, as we demonstrated in our prior study on glioblastoma metabolic symbiosis.

View Article and Find Full Text PDF

Although sulfur-bearing minerals are valuable resources, they pose significant environmental risks to river ecosystems by releasing hazardous leachate. Accurately tracing these sources is crucial but challenging due to overlapping chemical signatures and pollutant transport dynamics in river systems. This study investigates seasonal and spatial variations in sulfate (SO) and trace element contributions in mining districts of the upper Nakdong River basin, South Korea.

View Article and Find Full Text PDF

The complete tricarboxylic acid (TCA) cycle, comprising a series of 8 oxidative reactions, occurs in most eukaryotes in the mitochondria and in many prokaryotes. The net outcome of these 8 chemical reactions is the release of the reduced electron carriers NADH and FADH, water, and carbon dioxide. The parasites of the .

View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!