Protocols for Profiling of Metabolites and Metabolic Fluxes in Mammals

A special issue of Metabolites (ISSN 2218-1989). This special issue belongs to the section "Metabolomic Profiling Technology".

Deadline for manuscript submissions: closed (31 January 2019) | Viewed by 10091

Special Issue Editor


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Guest Editor
Department of Biochemistry and Molecular Biology, University of Southern Denmark, 5230 Odense, Denmark
Interests: coordination of lipid uptake; metabolism; and signalling in eukaryotes; lipid storage and energy metabolism in metazoans; ceramide/sphingolipid metabolism; dyslipidemia associated with obesity; diabetes and cardiovascular disease; application of metabolomics/lipidomics to identification of biomarkers; diagnosis; risk assessment

Special Issue Information

Dear Colleagues,

Global analyses and large-scale sequencing of genomes have led to two major shifts in the way scientific questions are approached. Firstly, the “omic” technical platforms have triggered a “high-throughput” revolution, which has allowed us to simultaneously determine hundreds to thousands of different molecular quantities. Secondly, we are increasingly aiming at understanding the importance of gene- and protein functions as part of metabolic networks. Therefore, quantitative analyses of the molecular quantities that define the activity of cellular networks are required. Metabolites are the end products of cellular biochemical processes, and their abundance is increasingly being considered as the ultimate response of biological systems to genetic, nutritional, or environmental changes. By analogy to the terms ‘genome’, ‘transcriptome’, and ‘proteome’, the set of metabolites present in a biological system comprises its ‘metabolome’. The use of isotope-labelled tracers can complement such data sets and allow the visualization of the dynamics of metabolic processes. As tracers are metabolized within tissues and cells, labelled isotopes become enriched in various metabolites, and this incorporation is a function of the label flux into and out of the metabolite pools. Thus, temporal isotopic labelling patterns can provide information on the ‘fluxome’ within tissues and cells. The new mass spectrometry-based technologies have provided the research community with the ability to assess changes in signalling networks and metabolites on a broad scale, providing a global perspective about how cells and organisms respond and adapt to, e.g., genetic, environmental, and nutritional changes, and therefore can be used to define cellular pathways, networks, and disease mechanisms.

The intention of this Special Issue of Metabolites is to provide a comprehensive collection of protocols used to globally profile metabolomes in mammalian biofluids and tissues, to identify and quantify selected groups of metabolites by targeted approaches, and, lastly, to chart the dynamics of metabolic fluxes. The issue will cover topics ranging from basic concepts of metabolomics, sample preparation, and analytical methodologies, to data interpretation and their applications in biomedicine. We anticipate that this issue will provide a useful resource not only to already established investigators with experience in metabolomics, but also to newcomers in the field, and may contribute to the standardization of metabolomics protocols used for profiling and quantification of metabolites in mammalian samples. We enthusiastically invite you to contribute to this Special Issue dedicated to “Protocols for Profiling of Metabolites and Metabolic Fluxes in Mammals”. We are confident that this issue will benefit the researchers who are interested and working in this field and become well cited in the future. Please use the Microsoft Word template (Protocol) to prepare your manuscript.

Prof. Dr. Nils Færgeman
Guest Editor

Manuscript Submission Information

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Keywords

  • Targeted metabolomics
  • Untargeted metabolomics
  • Mass spectrometry
  • LC–MS
  • GC–MS
  • Capillary electrophoresis
  • Ion mobility mass spectrometry

Published Papers (2 papers)

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19 pages, 4232 KiB  
Protocol
Efficient Extraction from Mice Feces for NMR Metabolomics Measurements with Special Emphasis on SCFAs
by Adrian Hauser, Philipp Eisenmann, Claudia Muhle-Goll, Burkhard Luy, Andreas Dötsch, Daniela Graf and Pavleta Tzvetkova
Metabolites 2019, 9(3), 55; https://doi.org/10.3390/metabo9030055 - 21 Mar 2019
Cited by 6 | Viewed by 5593
Abstract
Nuclear magnetic resonance (NMR) spectroscopy is one of the most promising methods for use in metabolomics studies as it is able to perform non targeted measurement of metabolites in a quantitative and non-destructive way. Sample preparation of liquid samples like urine or blood [...] Read more.
Nuclear magnetic resonance (NMR) spectroscopy is one of the most promising methods for use in metabolomics studies as it is able to perform non targeted measurement of metabolites in a quantitative and non-destructive way. Sample preparation of liquid samples like urine or blood serum is comparatively easy in NMR metabolomics, because mainly buffer and chemical shift reference substance are added. For solid samples like feces suitable extraction protocols need to be defined as initial step, where the exact protocol depends on sample type and features. Focusing on short chain fatty acids (SCFAs) in mice feces, we describe here a set of extraction protocols developed with the aim to suppress changes in metabolite composition within 24 h after extraction. Feces are obtained from mice fed on either standard rodent diet or high fat diet. The protocols presented in this manuscript are straightforward for application, and successfully minimize residual bacterial and enzymatic activities. Additionally, they are able to minimize the lipid background originating from the high fat diet. Full article
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11 pages, 2020 KiB  
Protocol
An UPLC-MS/MS Assay to Measure Glutathione as Marker for Oxidative Stress in Cultured Cells
by Katharina Herzog, Lodewijk IJlst, Arno G. van Cruchten, Carlo W.T. van Roermund, Wim Kulik, Ronald J. A. Wanders and Hans R. Waterham
Metabolites 2019, 9(3), 45; https://doi.org/10.3390/metabo9030045 - 5 Mar 2019
Cited by 20 | Viewed by 4077
Abstract
Oxidative stress plays a role in the onset and progression of a number of diseases, such as Alzheimer’s disease, diabetes and cancer, as well as ageing. Oxidative stress is caused by an increased production of reactive oxygen species and reduced antioxidant activity, resulting [...] Read more.
Oxidative stress plays a role in the onset and progression of a number of diseases, such as Alzheimer’s disease, diabetes and cancer, as well as ageing. Oxidative stress is caused by an increased production of reactive oxygen species and reduced antioxidant activity, resulting in the oxidation of glutathione. The ratio of reduced to oxidised glutathione is often used as a marker of the redox state in the cell. Whereas a variety of methods have been developed to measure glutathione in blood samples, methods to measure glutathione in cultured cells are scarce. Here we present a protocol to measure glutathione levels in cultured human and yeast cells using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC–MS/MS). Full article
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