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      Comparative metabolomics of muscle interstitium fluid in human trapezius myalgia: an in vivo microdialysis study

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          Abstract

          Purpose

          The mechanisms behind trapezius myalgia are unclear. Many hypotheses have been presented suggesting an altered metabolism in the muscle. Here, muscle microdialysate from healthy and myalgic muscle is analysed using metabolomics. Metabolomics analyse a vast number of metabolites, enabling a comprehensive explorative screening of the cellular processes in the muscle.

          Methods

          Microdialysate samples were obtained from the shoulder muscle of healthy and myalgic subjects that performed a work and stress test. Samples from the baseline period and from the recovery period were analysed using gas chromatography—mass spectrometry (GC–MS) together with multivariate analysis to detect differences in extracellular content of metabolites between groups. Systematic differences in metabolites between groups were identified using multivariate analysis and orthogonal partial least square discriminate analysis (OPLS-DA). A complementary Mann–Whitney U test of group difference in individual metabolites was also performed.

          Results

          A large number of metabolites were detected and identified in this screening study. At baseline, no systematic differences between groups were observed according to the OPLS-DA. However, two metabolites, l-leucine and pyroglutamic acid, were significantly more abundant in the myalgic muscle compared to the healthy muscle. In the recovery period, systematic difference in metabolites between the groups was observed according to the OPLS-DA. The groups differed in amino acids, fatty acids and carbohydrates. Myristic acid and putrescine were significantly more abundant and beta- d-glucopyranose was significantly less abundant in the myalgic muscle.

          Conclusion

          This study provides important information regarding the metabolite content, thereby presenting new clues regarding the pathophysiology of the myalgic muscle.

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          Most cited references73

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          Metabolomics: Current analytical platforms and methodologies

          (2005)
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            Extraction and GC/MS analysis of the human blood plasma metabolome.

            Analysis of the entire set of low molecular weight compounds (LMC), the metabolome, could provide deeper insights into mechanisms of disease and novel markers for diagnosis. In the investigation, we developed an extraction and derivatization protocol, using experimental design theory (design of experiment), for analyzing the human blood plasma metabolome by GC/MS. The protocol was optimized by evaluating the data for more than 500 resolved peaks using multivariate statistical tools including principal component analysis and partial least-squares projections to latent structures (PLS). The performance of five organic solvents (methanol, ethanol, acetonitrile, acetone, chloroform), singly and in combination, was investigated to optimize the LMC extraction. PLS analysis demonstrated that methanol extraction was particularly efficient and highly reproducible. The extraction and derivatization conditions were also optimized. Quantitative data for 32 endogenous compounds showed good precision and linearity. In addition, the determined amounts of eight selected compounds agreed well with analyses by independent methods in accredited laboratories, and most of the compounds could be detected at absolute levels of approximately 0.1 pmol injected, corresponding to plasma concentrations between 0.1 and 1 microM. The results suggest that the method could be usefully integrated into metabolomic studies for various purposes, e.g., for identifying biological markers related to diseases.
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              Microdialysis--principles and applications for studies in animals and man.

              Microdialysis is a technique for sampling the chemistry of the individual tissues and organs of the body, and is applicable to both animal and human studies. The basic principle is to mimic the function of a capillary blood vessel by perfusing a thin dialysis tube implanted into the tissue with a physiological liquid. The perfusate is analysed chemically and reflects the composition of the extracellular fluid with time due to the diffusion of substances back and forth over the membrane. Microdialysis is thus a technique whereby substances may be both recovered from and supplied to a tissue. The most important features of microdialysis are as follows: it samples the extracellular fluid, which is the origin of all blood chemistry; it samples continuously for hours or days without withdrawing blood; and it purifies the sample and simplifies chemical analysis by excluding large molecules from the perfusate. However, the latter feature renders the technique unsuitable for sampling large molecules such as proteins. The technique has been extensively used in the neurosciences to monitor neurotransmitter release, and is now finding application in monitoring of the chemistry of peripheral tissues in both animal and human studies.
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                Author and article information

                Contributors
                jhi@hig.se
                Journal
                Eur J Appl Physiol
                Eur. J. Appl. Physiol
                European Journal of Applied Physiology
                Springer Berlin Heidelberg (Berlin/Heidelberg )
                1439-6319
                1439-6327
                28 September 2013
                28 September 2013
                2013
                : 113
                : 2977-2989
                Affiliations
                [ ]Section for Anatomy, Department of Integrative Medical Biology, Umeå University, 901 87 Umeå, Sweden
                [ ]Department of Occupational and Public Health Sciences, Faculty of Health and Occupational Studies, Centre for Musculoskeletal Research, University of Gävle, 907 12 Umeå, Sweden
                [ ]Rehabilitation Medicine, Department of Medicine and Health Sciences (IMH), Faculty of Health Sciences, Pain and Rehabilitation Centre, Linköping University, County Council of Östergötland, 581 85 Linköping, Sweden
                [ ]Occupational and Environmental Medicine, Department of Clinical and Experimental Medicine, Faculty of Health Sciences, Centre of Occupational and Environmental Medicine, Linköping University, County Council of Östergötland, 581 85 Linköping, Sweden
                [ ]Institute of Stress Medicine, Carl Skottsbergs Gata 22B, 413 19 Gothenburg, Sweden
                [ ]Department of Chemistry, Faculty of Science and Technology, Umeå University, 901 85 Umeå, Sweden
                Author notes

                Communicated by Peter Krustrup.

                Article
                2716
                10.1007/s00421-013-2716-6
                3828502
                24078209
                8d78313b-d1e7-4e60-918f-e2c5d4d014c5
                © The Author(s) 2013

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited.

                History
                : 2 January 2013
                : 23 August 2013
                Categories
                Original Article
                Custom metadata
                © Springer-Verlag Berlin Heidelberg 2013

                Anatomy & Physiology
                metabolomics,trapezius myalgia,microdialysis,repetitive work,recovery,gc–ms,metabolites

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