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Chiara Mele Division of Endocrinology, Department of Translational Medicine, University of Piemonte Orientale, Novara, Italy
Division of General Medicine, Istituto Auxologico Italiano, IRCCS, S. Giuseppe Hospital, Piancavallo di Oggebbio (VB), Italy

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Maria Teresa Samà Division of Endocrinology, University Hospital ‘Maggiore della Carità’, Novara, Italy

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Alessandro Angelo Bisoffi Division of Endocrinology, Department of Translational Medicine, University of Piemonte Orientale, Novara, Italy

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Marina Caputo Division of Endocrinology, Department of Translational Medicine, University of Piemonte Orientale, Novara, Italy

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Valentina Bullara Division of Endocrinology, University Hospital ‘Maggiore della Carità’, Novara, Italy

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Stefania Mai Laboratory of Metabolic Research, Istituto Auxologico Italiano, IRCCS, S. Giuseppe Hospital, Piancavallo di Oggebbio (VB), Italy

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Gillian Elisabeth Walker Department of Health Sciences, University of Piemonte Orientale, Novara, Italy

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Flavia Prodam Department of Health Sciences, University of Piemonte Orientale, Novara, Italy

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Paolo Marzullo Division of Endocrinology, Department of Translational Medicine, University of Piemonte Orientale, Novara, Italy
Division of General Medicine, Istituto Auxologico Italiano, IRCCS, S. Giuseppe Hospital, Piancavallo di Oggebbio (VB), Italy

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Gianluca Aimaretti Division of Endocrinology, Department of Translational Medicine, University of Piemonte Orientale, Novara, Italy

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Loredana Pagano Division of Endocrinology, Department of Translational Medicine, University of Piemonte Orientale, Novara, Italy
Division of Endocrinology, Diabetology and Metabolism, Department of Medical Sciences, University of Turin, Turin, Italy

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assays, procedures were performed in accordance with the manufacturers’ instruction and the samples were analysed in duplicate. Serum leptin levels were assessed using a commercially available human ELISA kit (Mediagnost, Reutlinger, Germany). Intra

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Yao Ye Department of Obstetrics and Gynecology, University Hospital, LMU Munich, Munich, Germany

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Aurelia Vattai Department of Obstetrics and Gynecology, University Hospital, LMU Munich, Munich, Germany

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Nina Ditsch Department of Obstetrics and Gynecology, University Hospital, LMU Munich, Munich, Germany

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Christina Kuhn Department of Obstetrics and Gynecology, University Hospital, LMU Munich, Munich, Germany

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Martina Rahmeh Department of Obstetrics and Gynecology, University Hospital, LMU Munich, Munich, Germany

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Sven Mahner Department of Obstetrics and Gynecology, University Hospital, LMU Munich, Munich, Germany

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Myriam Ripphahn Walter Brendel Centre of Experimental Medicine, Department of Cardiovascular Physiology and Pathophysiology, Biomedical Center, Ludwig-Maximilians University of Munich, Munich, Germany

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Roland Immler Walter Brendel Centre of Experimental Medicine, Department of Cardiovascular Physiology and Pathophysiology, Biomedical Center, Ludwig-Maximilians University of Munich, Munich, Germany

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Markus Sperandio Walter Brendel Centre of Experimental Medicine, Department of Cardiovascular Physiology and Pathophysiology, Biomedical Center, Ludwig-Maximilians University of Munich, Munich, Germany

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Udo Jeschke Department of Obstetrics and Gynecology, University Hospital, LMU Munich, Munich, Germany

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Viktoria von Schönfeldt Department of Obstetrics and Gynecology, University Hospital, LMU Munich, Munich, Germany

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.8%, respectively. PAI-1 ELISA PAI-1 levels in the supernatants of JEG-3 and HTR-8/SVneo were measured utilizing commercially available ELISA kit (R&D system, DSE100). A standard curve was obtained in parallel to each assay and results converted into ng

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Jiaxi Li Department of Physiology, Xiangya School of Medicine, Central South University, Changsha, Hunan, China

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Pu Huang Department of Health Management Center, Changsha Central Hospital, University of South China, Changsha, Hunan, China

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Jing Xiong Department of Endocrinology, Xiangya Third Hospital, Central South University, Changsha, Hunan, China

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Xinyue Liang Department of Physiology, Xiangya School of Medicine, Central South University, Changsha, Hunan, China

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Mei Li Department of Physiology, Xiangya School of Medicine, Central South University, Changsha, Hunan, China

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Hao Ke Department of Physiology, Xiangya School of Medicine, Central South University, Changsha, Hunan, China

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Chunli Chen Department of Dermatology, Xiangya Third Hospital, Central South University, Changsha, Hunan, China

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Yang Han Department of Physiology, Xiangya School of Medicine, Central South University, Changsha, Hunan, China

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Yanhong Huang Department of Physiology, Xiangya School of Medicine, Central South University, Changsha, Hunan, China

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Yan Zhou Department of Physiology, Xiangya School of Medicine, Central South University, Changsha, Hunan, China

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Ziqiang Luo Department of Physiology, Xiangya School of Medicine, Central South University, Changsha, Hunan, China
Hunan Key Laboratory of Organ Fibrosis, Central South University, Changsha, Hunan, China

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Dandan Feng Department of Physiology, Xiangya School of Medicine, Central South University, Changsha, Hunan, China
Hunan Key Laboratory of Organ Fibrosis, Central South University, Changsha, Hunan, China

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Chen Chen School of Biomedical Sciences, University of Queensland, St Lucia, Brisbane, Queensland, Australia

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glucose, HbA1c, total cholesterol (TC), HDL-C, LDL-C, and triacylglycerols (TG). For insulin, ghrelin, and LEAP2 measurement, separate blood samples were collected without anticoagulation to obtain serum for ELISA assay. All blood samples were centrifuged

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Marta Fichna Department of Endocrinology, Metabolism and Internal Medicine, Poznan University of Medical Sciences, Poznan, Poland

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Piotr P Małecki Department of Endocrinology, Metabolism and Internal Medicine, Poznan University of Medical Sciences, Poznan, Poland

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Magdalena Żurawek Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland

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Katarzyna Furman Ludwik Perzyna Regional Hospital, Kalisz, Poland

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Bolesław Gębarski Regional Outpatient Medical Centre, Katowice, Poland

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Piotr Fichna Department of Paediatric Diabetes and Obesity, Poznan University of Medical Sciences, Poznan, Poland

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Marek Ruchała Department of Endocrinology, Metabolism and Internal Medicine, Poznan University of Medical Sciences, Poznan, Poland

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transporter-8 (aZnT8), and tyrosine phosphatase (aIA2) were detected by RIA- and ELISA-based commercial assays: 21-Hydroxylase Autoantibody ELISA Kit (RSR Ltd. Cardiff, UK), Anti-TPOn RIA kit, ThermoScientific (BRAHMS GmbH, Hennigsdorf, Germany), Anti-TGn RIA

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Bushra Shahida Department of Clinical Sciences, Genomics, Diabetes and Endocrinology, Lund University, Malmö, Sweden
Department of Diabetes & Endocrinology, Skåne University Hospital, Malmö, Sweden

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Tereza Planck Department of Clinical Sciences, Genomics, Diabetes and Endocrinology, Lund University, Malmö, Sweden
Department of Diabetes & Endocrinology, Skåne University Hospital, Malmö, Sweden

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Tania Singh Department of Clinical Sciences, Genomics, Diabetes and Endocrinology, Lund University, Malmö, Sweden

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Peter Åsman Department of Clinical Sciences Malmö, Ophthalmology, Lund University, Malmö, Sweden
Department of Ophthalmology, Skåne University Hospital, Malmö, Sweden

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Mikael Lantz Department of Clinical Sciences, Genomics, Diabetes and Endocrinology, Lund University, Malmö, Sweden
Department of Diabetes & Endocrinology, Skåne University Hospital, Malmö, Sweden

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-linked immunosorbent assay Cell culture supernatants were harvested, and the protein levels of PGE 2 , IL-1B, IL-6, and IgG were analyzed with commercially available ELISA kits (R & D Systems) according to the manufacturer’s guidelines. Cell culture for flow

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Hui Long Department of Assisted Reproduction, Shanghai Ninth People’s Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China

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Yanhong Nie Institute of Neuroscience, State Key Laboratory of Neuroscience, Key Laboratory of Primate Neurobiology, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China

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Li Wang Department of Assisted Reproduction, Shanghai Ninth People’s Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China

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Yong Lu Institute of Neuroscience, State Key Laboratory of Neuroscience, Key Laboratory of Primate Neurobiology, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China

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Yan Wang Institute of Neuroscience, State Key Laboratory of Neuroscience, Key Laboratory of Primate Neurobiology, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China

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Yijun Cai Institute of Neuroscience, State Key Laboratory of Neuroscience, Key Laboratory of Primate Neurobiology, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China

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Zhen Liu Institute of Neuroscience, State Key Laboratory of Neuroscience, Key Laboratory of Primate Neurobiology, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China

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Miaomiao Jia Department of Assisted Reproduction, Shanghai Ninth People’s Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China

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Qifeng Lyu Department of Assisted Reproduction, Shanghai Ninth People’s Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China

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Yanping Kuang Department of Assisted Reproduction, Shanghai Ninth People’s Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China

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Qiang Sun Institute of Neuroscience, State Key Laboratory of Neuroscience, Key Laboratory of Primate Neurobiology, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China

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aspiration. AMH assay Blood was withdrawn via an intravenous catheter and transferred into tubes for centrifugation. All serum was isolated from blood samples and was measured using an ELISA kit according to the manufacturer's instructions (AMH ELISA

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Kathryn L Gatford Robinson Research Institute, The University of Adelaide, Adelaide, Australia
Adelaide Medical School, The University of Adelaide, Adelaide, Australia

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Beverly S Muhlhausler FOOD plus Research Centre, School of Agriculture, Food and Wine, The University of Adelaide, Adelaide, Australia

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Lili Huang School of Biomedical Sciences, University of Queensland, St Lucia, Brisbane, Australia

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Pamela Su-Lin Sim FOOD plus Research Centre, School of Agriculture, Food and Wine, The University of Adelaide, Adelaide, Australia

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Claire T Roberts Robinson Research Institute, The University of Adelaide, Adelaide, Australia
Adelaide Medical School, The University of Adelaide, Adelaide, Australia

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Johannes D Velhuis Endocrine Research Unit, Mayo School of Graduate Medical Education, Center for Translational Science Activities, Mayo Clinic, Rochester, Minnesota, USA

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Chen Chen School of Biomedical Sciences, University of Queensland, St Lucia, Brisbane, Australia

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ELISA as described and validated previously ( 27 ). Within- and between-assay coefficients of variation were 2.3% and 14.7% ( n  = 12 plates) and all samples from a single mouse (serial samples for analysis of secretion pattern) were analysed on the same

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Esben Thyssen Vestergaard Medical Research Laboratories, Department of Endocrinology and Diabetes, Department of Pediatrics, Department of Endocrinology and Diabetes, Institute of Clinical Medicine, Aarhus University, Norrebrogade 44, DK-8000 Aarhus C, Denmark
Medical Research Laboratories, Department of Endocrinology and Diabetes, Department of Pediatrics, Department of Endocrinology and Diabetes, Institute of Clinical Medicine, Aarhus University, Norrebrogade 44, DK-8000 Aarhus C, Denmark
Medical Research Laboratories, Department of Endocrinology and Diabetes, Department of Pediatrics, Department of Endocrinology and Diabetes, Institute of Clinical Medicine, Aarhus University, Norrebrogade 44, DK-8000 Aarhus C, Denmark

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Morten B Krag Medical Research Laboratories, Department of Endocrinology and Diabetes, Department of Pediatrics, Department of Endocrinology and Diabetes, Institute of Clinical Medicine, Aarhus University, Norrebrogade 44, DK-8000 Aarhus C, Denmark

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Morten M Poulsen Medical Research Laboratories, Department of Endocrinology and Diabetes, Department of Pediatrics, Department of Endocrinology and Diabetes, Institute of Clinical Medicine, Aarhus University, Norrebrogade 44, DK-8000 Aarhus C, Denmark

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Steen B Pedersen Medical Research Laboratories, Department of Endocrinology and Diabetes, Department of Pediatrics, Department of Endocrinology and Diabetes, Institute of Clinical Medicine, Aarhus University, Norrebrogade 44, DK-8000 Aarhus C, Denmark

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Niels Moller Medical Research Laboratories, Department of Endocrinology and Diabetes, Department of Pediatrics, Department of Endocrinology and Diabetes, Institute of Clinical Medicine, Aarhus University, Norrebrogade 44, DK-8000 Aarhus C, Denmark
Medical Research Laboratories, Department of Endocrinology and Diabetes, Department of Pediatrics, Department of Endocrinology and Diabetes, Institute of Clinical Medicine, Aarhus University, Norrebrogade 44, DK-8000 Aarhus C, Denmark

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Jens Otto Lunde Jorgensen Medical Research Laboratories, Department of Endocrinology and Diabetes, Department of Pediatrics, Department of Endocrinology and Diabetes, Institute of Clinical Medicine, Aarhus University, Norrebrogade 44, DK-8000 Aarhus C, Denmark
Medical Research Laboratories, Department of Endocrinology and Diabetes, Department of Pediatrics, Department of Endocrinology and Diabetes, Institute of Clinical Medicine, Aarhus University, Norrebrogade 44, DK-8000 Aarhus C, Denmark

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Niels Jessen Medical Research Laboratories, Department of Endocrinology and Diabetes, Department of Pediatrics, Department of Endocrinology and Diabetes, Institute of Clinical Medicine, Aarhus University, Norrebrogade 44, DK-8000 Aarhus C, Denmark
Medical Research Laboratories, Department of Endocrinology and Diabetes, Department of Pediatrics, Department of Endocrinology and Diabetes, Institute of Clinical Medicine, Aarhus University, Norrebrogade 44, DK-8000 Aarhus C, Denmark

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levels were measured by an ELISA (EIA) from Alpco Diagnostics (Salem, NH, USA) with an intra-assay coefficient of variation (CV) of 5% and interassay CV of 9.7–9.8%. Plasma glucose was analyzed in duplicate using the glucose oxidase method (Beckman

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Angela Köninger
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Philippos Edimiris
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Laura Koch
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Antje Enekwe
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Claudia Lamina Department of Gynecology and Obstetrics, Division of Genetic Epidemiology, Vitateq Biotechnology GmbH, University of Duisburg-Essen, D-45122 Essen, Germany

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Sabine Kasimir-Bauer
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Rainer Kimmig
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Hans Dieplinger Department of Gynecology and Obstetrics, Division of Genetic Epidemiology, Vitateq Biotechnology GmbH, University of Duisburg-Essen, D-45122 Essen, Germany
Department of Gynecology and Obstetrics, Division of Genetic Epidemiology, Vitateq Biotechnology GmbH, University of Duisburg-Essen, D-45122 Essen, Germany

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-antibody sandwich ELISA using an affinity-purified biotinylated polyclonal anti-afamin antibody for coating 96-well streptavidin-bound microtiter plates and peroxidase-conjugated MAB N13 for detection (MicroCoat Biotechnologie GmbH, Bernried, Germany). Secondary

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Ram P Narayanan Vascular Research Group, School of Community Based Medicine, Centre for Integrated Genomic Medical Research, Cardiovascular Research Group, Endocrinology and Diabetes, Salford R&D, Department of Endocrinology and Diabetes, Faculty of Medical, Human and Life Sciences, The University of Manchester, Manchester M13 9PT, UK

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Bo Fu Vascular Research Group, School of Community Based Medicine, Centre for Integrated Genomic Medical Research, Cardiovascular Research Group, Endocrinology and Diabetes, Salford R&D, Department of Endocrinology and Diabetes, Faculty of Medical, Human and Life Sciences, The University of Manchester, Manchester M13 9PT, UK

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Adrian H Heald Vascular Research Group, School of Community Based Medicine, Centre for Integrated Genomic Medical Research, Cardiovascular Research Group, Endocrinology and Diabetes, Salford R&D, Department of Endocrinology and Diabetes, Faculty of Medical, Human and Life Sciences, The University of Manchester, Manchester M13 9PT, UK

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Kirk W Siddals Vascular Research Group, School of Community Based Medicine, Centre for Integrated Genomic Medical Research, Cardiovascular Research Group, Endocrinology and Diabetes, Salford R&D, Department of Endocrinology and Diabetes, Faculty of Medical, Human and Life Sciences, The University of Manchester, Manchester M13 9PT, UK

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Robert L Oliver Vascular Research Group, School of Community Based Medicine, Centre for Integrated Genomic Medical Research, Cardiovascular Research Group, Endocrinology and Diabetes, Salford R&D, Department of Endocrinology and Diabetes, Faculty of Medical, Human and Life Sciences, The University of Manchester, Manchester M13 9PT, UK

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Julie E Hudson Vascular Research Group, School of Community Based Medicine, Centre for Integrated Genomic Medical Research, Cardiovascular Research Group, Endocrinology and Diabetes, Salford R&D, Department of Endocrinology and Diabetes, Faculty of Medical, Human and Life Sciences, The University of Manchester, Manchester M13 9PT, UK

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Antony Payton Vascular Research Group, School of Community Based Medicine, Centre for Integrated Genomic Medical Research, Cardiovascular Research Group, Endocrinology and Diabetes, Salford R&D, Department of Endocrinology and Diabetes, Faculty of Medical, Human and Life Sciences, The University of Manchester, Manchester M13 9PT, UK

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Simon G Anderson Vascular Research Group, School of Community Based Medicine, Centre for Integrated Genomic Medical Research, Cardiovascular Research Group, Endocrinology and Diabetes, Salford R&D, Department of Endocrinology and Diabetes, Faculty of Medical, Human and Life Sciences, The University of Manchester, Manchester M13 9PT, UK

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Anne White Vascular Research Group, School of Community Based Medicine, Centre for Integrated Genomic Medical Research, Cardiovascular Research Group, Endocrinology and Diabetes, Salford R&D, Department of Endocrinology and Diabetes, Faculty of Medical, Human and Life Sciences, The University of Manchester, Manchester M13 9PT, UK

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William E R Ollier Vascular Research Group, School of Community Based Medicine, Centre for Integrated Genomic Medical Research, Cardiovascular Research Group, Endocrinology and Diabetes, Salford R&D, Department of Endocrinology and Diabetes, Faculty of Medical, Human and Life Sciences, The University of Manchester, Manchester M13 9PT, UK
Vascular Research Group, School of Community Based Medicine, Centre for Integrated Genomic Medical Research, Cardiovascular Research Group, Endocrinology and Diabetes, Salford R&D, Department of Endocrinology and Diabetes, Faculty of Medical, Human and Life Sciences, The University of Manchester, Manchester M13 9PT, UK

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J Martin Gibson Vascular Research Group, School of Community Based Medicine, Centre for Integrated Genomic Medical Research, Cardiovascular Research Group, Endocrinology and Diabetes, Salford R&D, Department of Endocrinology and Diabetes, Faculty of Medical, Human and Life Sciences, The University of Manchester, Manchester M13 9PT, UK
Vascular Research Group, School of Community Based Medicine, Centre for Integrated Genomic Medical Research, Cardiovascular Research Group, Endocrinology and Diabetes, Salford R&D, Department of Endocrinology and Diabetes, Faculty of Medical, Human and Life Sciences, The University of Manchester, Manchester M13 9PT, UK

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-assay coefficients of variation (CV) of <4.5 and <8.4% respectively. IGF2 was measured using an ELISA developed using antibodies that have been previously reported (11) . The analytical sensitivity of the assay was <10 ng/ml. The intra-assay and inter

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