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      The Bovine Metabolome

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          Abstract

          From an animal health perspective, relatively little is known about the typical or healthy ranges of concentrations for many metabolites in bovine biofluids and tissues. Here, we describe the results of a comprehensive, quantitative metabolomic characterization of six bovine biofluids and tissues, including serum, ruminal fluid, liver, Longissimus thoracis (LT) muscle, semimembranosus (SM) muscle, and testis tissues. Using nuclear magnetic resonance (NMR) spectroscopy, liquid chromatography–tandem mass spectrometry (LC–MS/MS), and inductively coupled plasma–mass spectrometry (ICP–MS), we were able to identify and quantify more than 145 metabolites in each of these biofluids/tissues. Combining these results with previous work done by our team on other bovine biofluids, as well as previously published literature values for other bovine tissues and biofluids, we were able to generate quantitative reference concentration data for 2100 unique metabolites across five different bovine biofluids and seven different tissues. These experimental data were combined with computer-aided, genome-scale metabolite inference techniques to add another 48,628 unique metabolites that are biochemically expected to be in bovine tissues or biofluids. Altogether, 51,801 unique metabolites were identified in this study. Detailed information on these 51,801 unique metabolites has been placed in a publicly available database called the Bovine Metabolome Database.

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          HMDB: a knowledgebase for the human metabolome

          The Human Metabolome Database (HMDB, http://www.hmdb.ca) is a richly annotated resource that is designed to address the broad needs of biochemists, clinical chemists, physicians, medical geneticists, nutritionists and members of the metabolomics community. Since its first release in 2007, the HMDB has been used to facilitate the research for nearly 100 published studies in metabolomics, clinical biochemistry and systems biology. The most recent release of HMDB (version 2.0) has been significantly expanded and enhanced over the previous release (version 1.0). In particular, the number of fully annotated metabolite entries has grown from 2180 to more than 6800 (a 300% increase), while the number of metabolites with biofluid or tissue concentration data has grown by a factor of five (from 883 to 4413). Similarly, the number of purified compounds with reference to NMR, LC-MS and GC-MS spectra has more than doubled (from 380 to more than 790 compounds). In addition to this significant expansion in database size, many new database searching tools and new data content has been added or enhanced. These include better algorithms for spectral searching and matching, more powerful chemical substructure searches, faster text searching software, as well as dedicated pathway searching tools and customized, clickable metabolic maps. Changes to the user-interface have also been implemented to accommodate future expansion and to make database navigation much easier. These improvements should make the HMDB much more useful to a much wider community of users.
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            Differential metabolomics reveals ophthalmic acid as an oxidative stress biomarker indicating hepatic glutathione consumption.

            Metabolomics is an emerging tool that can be used to gain insights into cellular and physiological responses. Here we present a metabolome differential display method based on capillary electrophoresis time-of-flight mass spectrometry to profile liver metabolites following acetaminophen-induced hepatotoxicity. We globally detected 1,859 peaks in mouse liver extracts and highlighted multiple changes in metabolite levels, including an activation of the ophthalmate biosynthesis pathway. We confirmed that ophthalmate was synthesized from 2-aminobutyrate through consecutive reactions with gamma-glutamylcysteine and glutathione synthetase. Changes in ophthalmate level in mouse serum and liver extracts were closely correlated and ophthalmate levels increased significantly in conjunction with glutathione consumption. Overall, our results provide a broad picture of hepatic metabolite changes following acetaminophen treatment. In addition, we specifically found that serum ophthalmate is a sensitive indicator of hepatic GSH depletion, and may be a new biomarker for oxidative stress. Our method can thus pinpoint specific metabolite changes and provide insights into the perturbation of metabolic pathways on a large scale and serve as a powerful new tool for discovering low molecular weight biomarkers.
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              Quantitative metabolomics using NMR

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                Author and article information

                Journal
                Metabolites
                Metabolites
                metabolites
                Metabolites
                MDPI
                2218-1989
                05 June 2020
                June 2020
                : 10
                : 6
                : 233
                Affiliations
                [1 ]Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, AB T6G 2P5, Canada; aidin@ 123456ualberta.ca (A.F.); cfitzsim@ 123456ualberta.ca (C.F.); lguan@ 123456ualberta.ca (L.L.G.)
                [2 ]Department of Biological Sciences, University of Alberta, Edmonton, AB T6G 2E9, Canada; rmandal@ 123456ualberta.ca (R.M.); pirimogh@ 123456ualberta.ca (H.P.-M.); jiamin3@ 123456ualberta.ca (J.Z.); aguo@ 123456ualberta.ca (A.G.); cbli@ 123456ualberta.ca (C.L.)
                [3 ]Agriculture and Agri-Food Canada, Edmonton, AB T6G 2P5, Canada
                [4 ]Department of Computing Science, University of Alberta, Edmonton, AB T6G 2E8, Canada
                Author notes
                Author information
                https://orcid.org/0000-0003-3021-7377
                https://orcid.org/0000-0001-9128-5029
                Article
                metabolites-10-00233
                10.3390/metabo10060233
                7345087
                32517015
                d6900a01-6c3a-42bf-9f6a-65e6b58f15f7
                © Her Majesty the Queen in Right of Canda as represented by the Minister of Agriculture and Agri-Food, 2020 and © author Carolyn Fitzsimmons, 2020.

                Submitted for possible open access publication under the terms and conditions of the Creative Commons Attribution(CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 08 May 2020
                : 02 June 2020
                Categories
                Article

                bovine,tissue,biofluid,nmr,lc–ms/ms,icp–ms,metabolite
                bovine, tissue, biofluid, nmr, lc–ms/ms, icp–ms, metabolite

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