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      Validation of standard operating procedures in a multicenter retrospective study to identify -omics biomarkers for chronic low back pain

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          Abstract

          Chronic low back pain (CLBP) is one of the most common medical conditions, ranking as the greatest contributor to global disability and accounting for huge societal costs based on the Global Burden of Disease 2010 study.

          Large genetic and -omics studies provide a promising avenue for the screening, development and validation of biomarkers useful for personalized diagnosis and treatment (precision medicine). Multicentre studies are needed for such an effort, and a standardized and homogeneous approach is vital for recruitment of large numbers of participants among different centres (clinical and laboratories) to obtain robust and reproducible results. To date, no validated standard operating procedures (SOPs) for genetic/-omics studies in chronic pain have been developed.

          In this study, we validated an SOP model that will be used in the multicentre (5 centres) retrospective “PainOmics” study, funded by the European Community in the 7th Framework Programme, which aims to develop new biomarkers for CLBP through three different -omics approaches: genomics, glycomics and activomics.

          The SOPs describe the specific procedures for ( 1) blood collection, ( 2) sample processing and storage, ( 3) shipping details and ( 4) cross-check testing and validation before assays that all the centres involved in the study have to follow.

          Multivariate analysis revealed the absolute specificity and homogeneity of the samples collected by the five centres for all genetics, glycomics and activomics analyses.

          The SOPs used in our multicenter study have been validated. Hence, they could represent an innovative tool for the correct management and collection of reliable samples in other large-omics-based multicenter studies.

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          Most cited references 18

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          The rising prevalence of chronic low back pain.

          National or state-level estimates on trends in the prevalence of chronic low back pain (LBP) are lacking. The objective of this study was to determine whether the prevalence of chronic LBP and the demographic, health-related, and health care-seeking characteristics of individuals with the condition have changed over the last 14 years. A cross-sectional, telephone survey of a representative sample of North Carolina households was conducted in 1992 and repeated in 2006. A total of 4437 households were contacted in 1992 and 5357 households in 2006 to identify noninstitutionalized adults 21 years or older with chronic (>3 months), impairing LBP or neck pain that limits daily activities. These individuals were interviewed in more detail about their health and health care seeking. The prevalence of chronic, impairing LBP rose significantly over the 14-year interval, from 3.9% (95% confidence interval [CI], 3.4%-4.4%) in 1992 to 10.2% (95% CI, 9.3%-11.0%) in 2006. Increases were seen for all adult age strata, in men and women, and in white and black races. Symptom severity and general health were similar for both years. The proportion of individuals who sought care from a health care provider in the past year increased from 73.1% (95% CI, 65.2%-79.8%) to 84.0% (95% CI, 80.8%-86.8%), while the mean number of visits to all health care providers were similar (19.5 [1992] vs 19.4 [2006]). The prevalence of chronic, impairing LBP has risen significantly in North Carolina, with continuing high levels of disability and health care use. A substantial portion of the rise in LBP care costs over the past 2 decades may be related to this rising prevalence.
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            Prognostic factors for duration of sick leave in patients sick listed with acute low back pain: a systematic review of the literature.

            The percentages of patients with acute low back pain (LBP) that go on to a chronic state varies between studies from 2% to 34%. In some of these cases low back pain leads to great costs. To evaluate the evidence for prognostic factors for return to work among workers sick listed with acute LBP. Systematic literature search with a quality assessment of studies, assessment of levels of evidence for all factors, and pooling of effect sizes. Inclusion of studies in the review was restricted to inception cohort studies of workers with LBP on sick leave for less than six weeks, with the outcome measured in absolute terms, relative terms, survival curve, or duration of sick leave. Of the studies, 18 publications (14 cohorts) fulfilled all inclusion criteria. One low quality study, four moderate quality studies, and nine high quality studies were identified; 79 prognostic factors were studied and grouped in eight categories for which the evidence was assessed. Specific LBP, higher disability levels, older age, female gender, more social dysfunction and more social isolation, heavier work, and receiving higher compensation were identified as predictors for a longer duration of sick leave. A history of LBP, job satisfaction, educational level, marital status, number of dependants, smoking, working more than 8 hour shifts, occupation, and size of industry or company do not influence duration of sick leave due to LBP. Many different constructs were measured to identify psychosocial predictors of long term sick leave, which made it impossible to determine the role of these factors.
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              High Throughput Isolation and Glycosylation Analysis of IgG–Variability and Heritability of the IgG Glycome in Three Isolated Human Populations*

              All immunoglobulin G molecules carry N-glycans, which modulate their biological activity. Changes in N-glycosylation of IgG associate with various diseases and affect the activity of therapeutic antibodies and intravenous immunoglobulins. We have developed a novel 96-well protein G monolithic plate and used it to rapidly isolate IgG from plasma of 2298 individuals from three isolated human populations. N-glycans were released by PNGase F, labeled with 2-aminobenzamide and analyzed by hydrophilic interaction chromatography with fluorescence detection. The majority of the structural features of the IgG glycome were consistent with previous studies, but sialylation was somewhat higher than reported previously. Sialylation was particularly prominent in core fucosylated glycans containing two galactose residues and bisecting GlcNAc where median sialylation level was nearly 80%. Very high variability between individuals was observed, approximately three times higher than in the total plasma glycome. For example, neutral IgG glycans without core fucose varied between 1.3 and 19%, a difference that significantly affects the effector functions of natural antibodies, predisposing or protecting individuals from particular diseases. Heritability of IgG glycans was generally between 30 and 50%. The individual's age was associated with a significant decrease in galactose and increase of bisecting GlcNAc, whereas other functional elements of IgG glycosylation did not change much with age. Gender was not an important predictor for any IgG glycan. An important observation is that competition between glycosyltransferases, which occurs in vitro, did not appear to be relevant in vivo, indicating that the final glycan structures are not a simple result of competing enzymatic activities, but a carefully regulated outcome designed to meet the prevailing physiological needs.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                1 May 2017
                2017
                : 12
                : 5
                Affiliations
                [1 ]Anesthesia, Critical Care and Pain Medicine Unit, Division of Surgical Sciences, Department of Medicine and Surgery, University of Parma, Parma, Italy
                [2 ]PainTherapy Service, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy
                [3 ]Institute of Epidemiology II, Research Unit of Molecular Epidemiology, Helmholtz Zentrum München, German Research Center for Environmental Health, Neuherberg, Germany
                [4 ]Genos, Glycoscience Research Laboratory, Zagreb, Croatia
                [5 ]Laboratory of Biochemistry and Genetics, Division of Pneumology, Department of Molecular Medicine, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy
                [6 ]School of Medical and Health Sciences, Edith Cowan University, Joondalup, Western Australia, Australia
                [7 ]IP Research Consulting SASU (PHOTEOMIX) - Rex de Chaussée, Noisy le Grand, France
                [8 ]Department of Twin Research and Genetic Epidemiology, King’s College London, London, United Kingdom
                [9 ]Department of Anesthesiology, Critical Care and Multidisciplinary Pain Center, Ziekenhuis Oost-Limburg, Genk, Belgium
                [10 ]St. Catherine Specialty Hospital, Zabok, Croatia
                [11 ]Eberly College of Science, The Pennsylvania State University, University Park, Pennsylvania, United States of America
                [12 ]University of Split School of Medicine, Split, Croatia
                [13 ]University of Osijek School of Medicine, Osijek, Croatia
                [14 ]Children's Hospital Srebrnjak, Zagreb, Croatia
                [15 ]PolyOmica, Groningen, The Netherlands
                [16 ]Carolinas Pain Institute, Winston-Salem, North Carolina, United States of America
                University of Hong Kong, HONG KONG
                Author notes

                Competing Interests: The authors stated that there are no conflicts of interest regarding the publication of this article. MA has received consultant fees or research grants from Grünenthal, Angelini, Mundipharma, MSD and Care Fusion. GL has multiple patents in the field of glycoscience issued. IKP and JMD are scientific directors of Photeomix, the commercial name of IP Research Consulting SAS, who have ongoing patent applications and trademarks related to Activomics. YA is a director and co-owner of Maatschap PolyOmica, which provides (consulting) services in the area of (statistical) (gen)omics. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

                • Conceptualization: CD MA MDG.

                • Data curation: CD.

                • Formal analysis: YA.

                • Funding acquisition: MA.

                • Investigation: CD.

                • Methodology: CD MDG YA.

                • Project administration: CD.

                • Resources: CD MDG CG JM IG GL LD WW MS IKP JMD FW JVZ DP YA LK MA.

                • Supervision: MA CG GL WW IKP FW JVZ DP.

                • Validation: CD MDG CG JM IG GL IKP JMD.

                • Visualization: CD MDG CG JM IG GL LD WW MS IKP JMD FW JVZ DP YA LK MA.

                • Writing – original draft: CD.

                • Writing – review & editing: CD MDG CG JM IG GL LD WW MS IKP JMD FW JVZ DP YA LK MA.

                ¶ Membership of the PainOmics Group is provided in the Acknowledgments.

                Article
                PONE-D-16-27833
                10.1371/journal.pone.0176372
                5411039
                28459826
                © 2017 Dagostino et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                Page count
                Figures: 6, Tables: 1, Pages: 13
                Product
                Funding
                Funded by: Seventh Framework Program of the European Community for Research, Technological Development and Demonstration Activities - (FP7)
                Award ID: 602736
                The present study is not funded by Industry or any other commercial sponsors. This trial is funded academic research; it is supported by funding from the European Commission FP7 Project in the context of the THEME [HEALTH.2013.2.2.1-5 - Understanding and controlling pain] named “PainOmics” (Grant agreement no: 602736). The funds received from the European grant provided support in the form of salaries for authors (CD, MDG, GL, JMD, YA) but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section.
                Categories
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