ࡱ>  ?B456789:;<=>ubjbjneem-)))))48)8)8)h)l +|8)DB11111222vDxDxDxDxDxDxD$9GIlD)22222D))11D>>>2 )1)1B>2vD>>@A1P7* <@BD0D@,WJ<PWJAAWJ)A22>22222DD>222D2222WJ222222222 ':Supplementary Information for Metabolic effects of a 13-weeks lifestyle intervention in older adults: The Growing Old Together Study Ondine van de Rest, PhD#1, Bianca A.M. Schutte, MSc#2, Joris Deelen, PhD#2, Stephanie A.M. Stassen, MSc3, Erik B. van den Akker, PhD2,4, Diana van Heemst, PhD 3, Petra Dibbets-Schneider, BSc5, Regina. A. van Dipten-van der Veen, BSc1, Milou Kelderman, BSc1, Thomas Hankemeier, PhD6, Simon P. Mooijaart, PhD 3, Jeroen van der Grond, PhD 5, Jeanine J. Houwing-Duistermaat, PhD 7, Marian Beekman, PhD 2, Edith J.M. Feskens, PhD 1, P. Eline Slagboom, PhD2 # These authors contributed equally to this work Division of Human Nutrition, Wageningen University, PO Box 8129, 6700 EV Wageningen, The Netherlands. Department of Molecular Epidemiology, Leiden University Medical Center, PO Box 9600, 2300 RC Leiden, The Netherlands. Department of Gerontology and Geriatrics, Leiden University Medical Center, PO Box 9600, 2300 RC Leiden, The Netherlands The Delft Bioinformatics Lab, Delft University of Technology, Mekelweg 4, 2628 CD Delft, The Netherlands Department of Radiology, Leiden University Medical Center, PO Box 9600, 2300 RC Leiden, The Netherlands Division of Analytical Biosciences, Leiden Academic Centre for Drug Research, Leiden University, Leiden 2300 RA, The Netherlands Suplementary Tables Supplementary Table 1A. Baseline characteristics of parameters of body composition, health and functioning, and diagnostic measurements. CharacteristicnLongevity family membersnControlsP-valueaWomen, n (%)39 (43.3)42 (56.8)Age, mean (SD) [range], years9063.4 (5.4) [49.1-75.1]7462.4 (6.1) [46.7-73.5]0.24Body composition, mean (SD) [range]Weight, kg8979.8 (9.6) [62.5-105.7]7379.0 (10.2) [60.5-102.4]0.74Men5084.3 (8.0) [67.2-105.7]3185.4 (8.1) [70.1-102.4]0.39Women3974.1 (8.4) [62.5-95.4]4274.1 (8.9) [60.5-100.4]0.73BMI, kg/m28927.0 (2.6) [22.9-34.2]7326.9 (2.4) [22.9-33.5]0.77Waist circumference, cm9096.2 (7.9) [74-122]7496.1 (8.2) [77-112]0.47Men5198.1 (7.4) [80-122]32100.1 (6.4) [89-112]0.26Women3993.6 (7.9) [74-112]4293.0 (8.1) [77-111]0.63Body fat, %8929.6 (8.8) [11.6-48.3]7330.9 (7.0) [17.1-51.0]0.98Men5023.7 (5.7) [11.6-38.7]3125.5 (4.7) [17.1-35.0]0.17Women3937.1 (5.8) [22.9-48.3]4234.9 (5.6) [21.0-51.0]0.13Fat free mass, kg28956.3 (10.1) [38.5-78.6]7454.6 (10.1) [37.0-81.6]0.58Men5064.1 (5.6) [49.5-78.6]3163.6 (7.5) [46.9-81.6]0.98Women3946.3 (3.9) [38.5-54.7]4248.0 (5.9) [37.0-65.5]0.30Health and functioning, mean (SD) [range]Systolic blood pressure, mmHgb65135.4 (15.9) [111-196]48137.8 (17.1) [101-173]0.99Diastolic blood pressure, mmHgb6583.5 (7.4) [64-101]4884.7 (9.2) [65-108]0.28REE, kcal/day681,186 (178) [890-1,680]581,189 (163) [871-1,680]0.42Men381,294 (150) [940-1,680]271,282 (160) [1,083-1,680]0.93Women301,048 (96) [890-1,300]311,108 (117) [872-1,349]0.04Handgrip strength, kg8943.2 (12.1) [21-67]7140.8 (10.3) [21-78]0.82Men5052.3 (7.5) [36-67]3049.5 (7.5) [39-78]0.20Women3931.6 (4.8) [21-42]4134.4 (6.6) [21-54]0.18Physical functioning9011.5 (0.6) [10-12]7311.6 (0.7) [10-12]0.31Physical quality of life8852.4 (5.9) [26.7-62.8]7252.9 (6.2) [26.8-63.0]0.61Men5152.7 (4.8) [39.0-58.9]3153.0 (4.2) [37.5-56.8]0.72Women3751.9 (7.2) [26.7-62.9]4152.7 (7.5) [26.8-63.0]0.78Mental quality of life8854.5 (6.7) [27.6-63.7]7253.6 (8.2) [19.4-67.0]0.71Men5156.2 (4.9) [41.5-63.7]3155.6 (5.7) [29.0-62.9]0.56Women3752.3 (8.1) [27.6-60.8]4152.1 (9.4) [19.4-67.0]0.65FRS, %908.3 (5.0) [1-20]748.9 (7.3) [1-25]<0.001Men5111.4 (3.6) [6-20]3216.2 (4.6) [10-25]<0.001Women394.3 (3.4) [1-17]423.3 (2.2) [1-11]0.67Medication use, n (%)Lipid-lowering agent9011 (12.2)7418 (24.3)0.03Antihypertensive agent9023 (25.6)7426 (35.1)0.22Diagnostic measurements, mean (SD) [range]Fasting glucose, mmol/L905.0 (0.5) [3.6-6.5]745.0 (0.6) [4.0-7.6]0.88Fasting insulin, mU/Lc909.4 (5.1) [2.0-29.6]749.0 (3.9) [2.0-22.6]0.84HOMA-IR881.2 (0.6) [0.4-3.8]721.2 (0.5) [0.4-2.7]0.72Total cholesterol, mmol/Ld795.5 (1.0) [3.3-8.6]565.5 (1.0) [3.2-8.0]0.58HDL cholesterol, mmol/Ld791.6 (0.4) [0.6-3.1]561.4 (0.4) [0.6-2.3]0.001Men431.4 (0.3) [1.0-2.0]231.1 (0.2) [0.6-1.6]<0.001Women361.7 (0.5) [0.6-3.1]331.6 (0.3) [1.2-2.3]0.39LDL cholesterol, mmol/Ld793.5 (0.8) [1.8-6.4]563.4 (0.9) [1.6-6.0]0.51Triglycerides, mmol/Lc,d791.3 (0.5) [0.6-3.0]561.2 (0.6) [0.5-4.1]0.13fT3, pmol/L904.5 (0.4) [3.3-5.5]744.6 (0.4) [3.5-6.2]0.54fT4, pmol/L9015.0 (1.8) [10.5-20.3]7414.7 (1.6) [10.5-19.3]0.21TSH, mU/Lc902.6 (1.6) [0.3-9.8]743.1 (3.0) [0.2-25.0]0.32DHEAS, nmol/Lc902.9 (1.8) [0.5-8.6]743.3 (1.6) [0.7-6.8]0.006Men513.6 (2.0) [0.6-8.6]323.8 (1.4) [1.5-6.8]0.12Women392.0 (1.2) [0.5-5.2]422.8 (1.6) [0.7-6.7]0.03Leptin, g/Lc9013.5 (11.5) [1.2-61.7]7414.4 (9.2) [2.3-50.3]0.52Men516.7 (3.8) [1.2-17.2]327.6 (3.5) [2.3-18.1]0.15Women3922.5 (12.2) [4.5-61.7]4219.7 (8.9) [6.5-50.3]0.35Adiponectin, mg/Lc9010.6 (5.2) [2.2-28.1]7410.1 (5.5) [2.7-25.6]0.04Men518.1 (3.5) [2.2-19.3]326.8 (3.0) [2.8-15.2]0.08Women3913.9 (5.2) [5.6-28.1]4212.7 (5.6) [2.7-25.6]0.30IGF-1, nmol/L9020.0 (4.8) [7.8-31.9]7420.0 (4.8) [8.4-43.0]0.57Men5120.8 (4.7) [7.8-31.9]3221.6 (5.1) [10.7-31.7]0.41Women3919.0 (4.6) [11.4-30.9]4219.3 (6.4) [8.4-43.0]0.88IGFBP-3, mg/L904.0 (0.9) [1.7-7.7]744.0 (0.9) [2.2-6.7]0.34IGF-1:IGFBP-3900.14 (0.03) [0.06-0.26]740.14 (0.03) [0.07-0.23]0.23Men510.15 (0.03) [0.07-0.26]320.15 (0.04) [0.10-0.23]0.68Women390.12 (0.02) [0.06-0.17]420.13 (0.03) [0.07-0.20]0.33CRP (high-sensitivity), mg/Lc901.9 (2.8) [0.2-18.8]742.3 (3.6) [0.2-20.0]0.23 a P value refers to difference between longevity family members and controls. b Individuals using antihypertensive agents were removed before analysis. c Natural log transformed parameter was used for analysis. d Individuals using lipid-lowering agents were removed before analysis. Parameters were analysed separately in men and women if there was a significant gender-difference at baseline. BMI, body mass index; REE, resting energy expenditure; FRS, Framingham risk score; HOMA-IR, homeostatic model assessment - insulin resistance; HDL, high density lipoprotein; LDL, low density lipoprotein; fT3, free triiodothyronine; fT4, free thyroxine; TSH, thyroid stimulating hormone; DHEAS, dehydroepiandrosterone-sulfate; IGF-1, insuline-like growth factor 1; IGFBP-3, insulin-like growth factor binding protein 3; CRP, C-reactive protein.. Supplementary Table 1B. Baseline characteristics of 1H-NMR metabolites. Characteristic, mean (SD) [range]nLongevity family membersnControlsP-valueaAmino acidsAlanine, mmol/L900.45 (0.05) [0.34-0.62]720.46 (0.06) [0.33-0.63]0.64Men510.44 (0.05) [0.35-0.58]300.43 (0.05) [0.33-0.56]0.52Women390.46 (0.06) [0.34-0.62]420.47 (0.06) [0.36-0.63]0.33Glutamine, mmol/L900.52 (0.06) [0.39-0.74]730.49 (0.05) [0.36-0.62]0.01Glycine, mmol/L900.29 (0.06) [0.20-0.56]730.29 (0.06) [0.16-0.54]0.30Men510.27 (0.04) [0.20-0.40]310.26 (0.04) [0.16-0.39]0.15Women390.32 (0.07) [0.24-0.56]420.32 (0.07) [0.21-0.54]0.94Histidine, mmol/L900.06 (0.01) [0.04-0.09]730.06 (0.01) [0.03-0.08]0.06Branched-chain amino acidsIsoleucine, mmol/L900.06 (0.02) [0.04-0.14]720.06 (0.02) [0.03-0.11]0.40Men510.06 (0.01) [0.04-0.10]300.07 (0.01) [0.05-0.11]0.09Women390.06 (0.02) [0.04-0.14]420.06 (0.01) [0.03-0.11]0.63Leucine, mmol/L900.09 (0.01) [0.05-0.13]720.09 (0.01) [0.06-0.14]0.64Men510.09 (0.01) [0.07-0.12]300.09 (0.01) [0.07-0.12]0.46Women390.08 (0.01) [0.05-0.13]420.08 (0.01) [0.06-0.14]0.78Valine, mmol/L890.20 (0.03) [0.12-0.28]720.20 (0.03) [0.15-0.34]0.44Aromatic amino acidsPhenylalanine, mmol/L900.08 (0.01) [0.06-0.11]730.08 (0.01) [0.04-0.10]0.72Tyrosine, mmol/L900.06 (0.01) [0.04-0.08]730.06 (0.01) [0.02-0.09]0.48Glycolysis-related metabolitesGlucose, mmol/L904.26 (0.48) [3.02-6.51]734.22 (0.50) [2.73-6.56]0.87Lactate, mmol/L901.84 (0.33) [1.22-3.02]731.84 (0.37) [0.71-2.78]0.89Pyruvate, mmol/L890.08 (0.03) [0.04-0.17]730.09 (0.02) [0.04-0.17]0.46Citrate, mmol/L900.10 (0.02) [0.06-0.15]730.10 (0.02) [0.04-0.18]0.12Glycerol, mmol/L880.08 (0.03) [0.04-0.20]730.08 (0.02) [0.03-0.14]0.41Men490.07 (0.03) [0.04-0.20]310.07 (0.02) [0.04-0.12]0.62Women390.09 (0.03) [0.04-0.15]420.08 (0.02) [0.03-0.14]0.09Ketone bodiesAcetate, mmol/L900.05 (0.09) [0.03-0.77]730.04 (0.01) [0.02-0.07]0.11Acetoacetate, mmol/L900.04 (0.02) [0.01-0.15]730.03 (0.01) [0.00-0.10]0.59Men510.04 (0.02) [0.02-0.15]310.04 (0.02) [0.00-0.10]0.50Women390.03 (0.01) [0.01-0.07]420.03 (0.01) [0.01-0.07]0.873-hydroxybutyrate, mmol/L880.09 (0.02) [0.04-0.16]730.09 (0.03) [0.03-0.19]0.88Fatty acidsbTotal fatty acids, mmol/L7612.11 (2.46) [8.55-23.03]5512.42 (2.02) [8.70-17.64]0.48Omega-3 fatty acids, mmol/L760.44 (0.11) [0.20-0.72]550.46 (0.11) [0.28-0.69]0.64Docosahexaenoic acid (DHA), mmol/L760.16 (0.05) [0.06-0.27]550.16 (0.04) [0.10-0.26]0.77Omega-6 fatty acids, mmol/L764.18 (0.72) [3.09-6.21]554.25 (0.65) [3.02-6.34]0.74Linoleic acid, mmol/L763.37 (0.58) [2.49-5.20]553.44 (0.55) [2.36-5.13]0.66Polyunsaturated fatty acids, mmol/L764.62 (0.79) [3.49-6.83]554.71 (0.71) [3.42-7.03]0.72Monounsaturated fatty acids, mmol/L762.76 (0.83) [1.54-6.49]552.85 (0.68) [1.80-5.35]0.35Saturated fatty acids, mmol/L764.73 (0.99) [3.28-9.71]554.87 (0.84) [3.35-6.93]0.46Fatty acid chain length7617.32 (0.34) [16.74-18.50]5517.38 (0.31) [16.66-18.20]0.46Fatty acid ratios, relative to total fatty acidsbOmega-3 fatty acids, %763.68 (0.64) [2.28-5.47]553.71 (0.77) [2.15-5.77]0.65Docosahexaenoic acid (DHA), %761.29 (0.31) [0.73-2.33]551.30 (0.32) [0.62-2.03]0.57Omega-6 fatty acids, %7634.75 (2.86) [26.55-41.19]5534.38 (2.36) [28.29-39.22]0.36Linoleic acid, %7628.08 (2.69) [20.56-35.62]5527.84 (2.40) [21.81-34.00]0.57Polyunsaturated fatty acids, %7638.43 (2.88) [29.65-43.47]5538.08 (2.59) [30.43-43.80]0.33Monounsaturated fatty acids, %7622.48 (2.60) [17.82-30.75]5522.77 (2.78) [18.89-30.91]0.25Saturated fatty acids, %7639.09 (1.75) [35.45-43.55]5539.15 (1.82) [35.19-43.52]0.97Unsaturation degree, double bonds per fatty acids761.19 (0.05) [1.03-1.29]551.19 (0.05) [1.02-1.31]0.57Fluid balanceCreatinine, mmol/L900.06 (0.01) [0.03-0.09]730.06 (0.01) [0.04-0.12]0.18Men510.07 (0.01) [0.03-0.09]310.07 (0.01) [0.04-0.12]0.09Women390.05 (0.01) [0.04-0.07]420.05 (0.01) [0.04-0.07]0.90Albumin, signal area900.09 (0.00) [0.08-0.10]730.09 (0.00) [0.08-0.10]0.91InflammationGlycoprotein acetyls, mainly a1-acid glycoprotein, mmol/L901.21 (0.20) [0.91-2.15]731.25 (0.18) [0.88-1.90]0.11Men511.17 (0.15) [0.91-1.54]311.31 (0.18) [0.99-1.90]<0.001Women391.26 (0.24) [0.97-2.15]421.20 (0.17) [0.88-1.57]0.25ApolipoproteinsbApolipoprotein B, g/L790.94 (0.23) [0.56-1.94]550.99 (0.18) [0.61-1.51]0.07Apolipoprotein A1, g/L791.60 (0.17) [1.21-2.07]551.55 (0.21) [1.08-2.10]<0.001Men431.53 (0.15) [1.21-1.87]221.39 (0.13) [1.08-1.57]<0.001Women361.68 (0.15) [1.43-2.07]331.66 (0.18) [1.28-2.10]0.68LipidsbTriglycerides, mmol/L791.16 (0.65) [0.41-4.88]551.26 (0.50) [0.63-3.16]0.03Men431.13 (0.40) [0.59-2.09]221.55 (0.56) [0.81-3.16]<0.001Women361.20 (0.86) [0.41-4.88]331.07 (0.35) [0.63-2.02]0.88Phosphoglycerides, mmol/L762.14 (0.32) [1.54-3.01]552.16 (0.37) [1.43-2.92]0.55Men422.06 (0.30) [1.54-2.62]221.97 (0.30) [1.43-2.45]0.33Women342.24 (0.32) [1.69-3.01]332.29 (0.36) [1.60-2.92]0.55Cholines, mmol/L762.55 (0.33) [1.93-3.30]552.57 (0.40) [1.68-3.42]0.45Men422.45 (0.34) [1.93-3.18]222.35 (0.32) [1.68-2.91]0.32Women342.67 (0.29) [2.07-3.30]332.71 (0.38) [2.00-3.42]0.61Sphingomyelins, mmol/L760.51 (0.09) [0.33-0.79]550.51 (0.08) [0.32-0.69]0.22Men420.48 (0.09) [0.33-0.79]220.47 (0.07) [0.32-0.62]0.55Women340.55 (0.07) [0.41-0.71]330.54 (0.08) [0.39-0.69]0.81Total lipid concentrations in lipoprotein subclassesbExtremely large VLDL, mmol/L730.03 (0.02) [0.00-0.14]520.03 (0.02) [0.00-0.11]0.12Very large VLDL, mmol/L 680.06 (0.07) [0.00-0.41]490.06 (0.06) [0.01-0.31]0.08Large VLDL, mmol/L730.22 (0.23) [0.02-1.56]530.23 (0.20) [0.03-1.09]0.42Medium VLDL, mmol/L 790.48 (0.37) [0.09-2.64]550.52 (0.30) [0.17-1.67]0.06Small VLDL, mmol/L 790.61 (0.26) [0.16-1.90]550.67 (0.20) [0.31-1.06]0.01Very small VLDL, mmol/L790.58 (0.16) [0.24-1.06]550.63 (0.14) [0.34-1.01]0.03IDL, mmol/L 791.28 (0.30) [0.76-2.22]551.35 (0.28) [0.77-2.24]0.14Large LDL, mmol/L 791.47 (0.35) [0.87-2.53]551.55 (0.33) [0.82-2.57]0.19Medium LDL, mmol/L 790.84 (0.21) [0.46-1.43]550.88 (0.19) [0.44-1.47]0.20Small LDL, mmol/L 790.54 (0.13) [0.31-0.89]550.56 (0.12) [0.30-0.93]0.30Very large HDL, mmol/L 790.48 (0.21) [0.10-1.14]550.42 (0.21) [0.11-0.96]0.001Men430.41 (0.16) [0.10-0.92]220.26 (0.10) [0.11-0.56]<0.001Women360.55 (0.23) [0.13-1.14]330.53 (0.20) [0.18-0.96]0.95Large HDL, mmol/L750.80 (0.29) [0.25-1.73]510.73 (0.34) [0.23-1.55]<0.001Men410.68 (0.24) [0.25-1.15]180.45 (0.16) [0.23-0.83]<0.001Women340.95 (0.29) [0.54-1.73]330.89 (0.31) [0.34-1.55]0.29Medium HDL, mmol/L 790.95 (0.16) [0.66-1.44]550.93 (0.20) [0.51-1.38]0.005Men430.91 (0.15) [0.66-1.44]220.79 (0.15) [0.51-1.10]0.003Women361.01 (0.15) [0.70-1.30]331.02 (0.17) [0.72-1.38]0.88Small HDL, mmol/L 791.15 (0.11) [0.92-1.42]551.15 (0.10) [0.95-1.38]0.70Men431.15 (0.10) [0.92-1.41]221.12 (0.09) [0.95-1.31]0.30Women361.15 (0.13) [0.95-1.42]331.16 (0.10) [0.97-1.38]0.48Lipoprotein particle sizebVLDL diameter, nm7935.76 (1.11) [33.87-39.22]5535.87 (1.26) [34.26-39.69]0.28LDL diameter, nm7923.58 (0.11) [23.33-23.83]5523.60 (0.10) [23.43-23.81]0.30HDL diameter, nm799.97 (0.24) [9.46-10.61]559.90 (0.27) [9.39-10.47]0.001Men439.90 (0.20) [9.46-10.41]229.67 (0.17) [9.39-10.08]<0.001Women3610.07 (0.25) [9.48-10.61]3310.04 (0.22) [9.64-10.47]0.62CholesterolbTotal cholesterol, mmol/L795.05 (0.90) [3.64-7.70]555.16 (0.91) [2.98-7.95]0.77VLDL cholesterol, mmol/L790.76 (0.33) [0.22-2.27]550.83 (0.24) [0.36-1.38]0.02IDL cholesterol, mmol/L790.82 (0.20) [0.48-1.47]550.86 (0.19) [0.45-1.48]0.23LDL cholesterol, mmol/L791.93 (0.51) [1.04-3.45]552.02 (0.47) [0.98-3.52]0.22HDL cholesterol, mmol/L791.55 (0.32) [0.79-2.49]551.45 (0.38) [0.68-2.35]<0.001Men431.44 (0.25) [0.96-1.96]221.14 (0.22) [0.68-1.51]<0.001Women361.68 (0.35) [0.79-2.49]331.66 (0.31) [1.08-2.35]0.97HDL2 cholesterol, mmol/L791.04 (0.30) [0.28-1.94]550.94 (0.34) [0.25-1.77]<0.001Men430.94 (0.24) [0.48-1.44]220.66 (0.20) [0.25-0.98]<0.001Women361.16 (0.32) [0.28-1.94]331.13 (0.28) [0.61-1.77]0.99HDL3 cholesterol, mmol/L790.51 (0.03) [0.42-0.60]550.51 (0.04) [0.42-0.59]0.03Men430.51 (0.03) [0.43-0.60]220.48 (0.03) [0.42-0.55]0.001Women360.52 (0.04) [0.42-0.60]330.53 (0.03) [0.47-0.59]0.33 a P value refers to difference between longevity family members and controls. b Individuals using lipid-lowering agents were removed before analysis. All metabolite concentrations were natural log-transformed and scaled to standard deviation units before analysis. Parameters were analysed separately in men and women if there was a significant gender-difference at baseline. VLDL, very low density lipoprotein; IDL, intermediate density lipoprotein; LDL, low density lipoprotein; HDL, high density lipoprotein. Supplementary Table 2A. Effects of the intervention on parameters of body composition, health and functioning, and diagnostic measurements. UnadjustedWeight loss adjustedCharacteristic, mean (SE)nDifferenceP-valueaDifferenceP-valuebBody compositionWeight, kg161-3.34 (0.18)<0.001Men80-3.42 (0.27)<0.001Women81-3.25 (0.23)<0.001BMI, kg/m2161-1.13 (0.06)<0.001Waist circumference, cm163-4.3 (0.4)<0.001Men82-4.4 (0.6)<0.001Women81-4.2 (0.6)<0.001Body fat, %161-2.26 (0.16)<0.001Men80-2.22 (0.23)<0.001Women81-2.29 (0.21)<0.001Fat free mass, kg2161-0.67 (0.10)<0.001Men80-0.83 (0.16)<0.001Women81-0.51 (0.13)<0.001Health and functioningSystolic blood pressure, mmHgc113-4.33 (0.98)<0.001-2.74 (1.12)0.01Diastolic blood pressure, mmHgc113-1.66 (0.61)0.007-0.52 (0.68)0.44REE, kcal/day126-49.2 (8.0)<0.001-20.2 (8.3)0.02Men65-46.59 (11.76)<0.001-8.09 (12.41)0.51Women61-51.94 (10.79)<0.001-28.42 (11.28)0.01Handgrip strength, kg1530.38 (0.32)0.25Men760.24 (0.53)0.65Women770.51 (0.38)0.18Physical functioning1590.14 (0.05)0.010.11 (0.06)0.04Physical quality of life157-0.18 (0.61)0.77Men82-0.72 (0.83)0.39Women750.42 (0.92)0.65Mental quality of life1570.9 (0.70)0.19Men82-1.13 (0.84)0.18Women753.13 (1.12)0.013.13 (1.16)0.007FRS, %163-0.51 (0.23)0.03-0.09 (0.24)0.73Men82-0.65 (0.43)0.13Women81-0.37 (0.15)0.01-0.15 (0.17)0.38Diagnostic measurementsFasting glucose, mmol/L163-0.06 (0.04)0.16Fasting insulin, mU/Ld163-0.05 (0.03)0.040.04 (0.03)0.21HOMA-IR153-0.03 (0.03)0.33Total cholesterol, mmol/Le135-0.29 (0.06)<0.001-0.28 (0.06)<0.001HDL cholesterol, mmol/Le135-0.01 (0.02)0.49Men660.04 (0.02)0.11Women69-0.06 (0.03)0.02-0.08 (0.03)0.02LDL cholesterol, mmol/Le135-0.26 (0.05)<0.001-0.23 (0.05)<0.001Triglycerides, mmol/Ld,e135-0.04 (0.03)0.11fT3, pmol/L163-0.14 (0.03)<0.001-0.12 (0.03)<0.001fT4, pmol/L163-0.07 (0.09)0.44TSH, mU/Le163-0.04 (0.03)0.17DHEAS, nmol/Ld163-0.02 (0.01)0.20Men82-0.01 (0.02)0.47Women81-0.02 (0.02)0.28Leptin, g/Ld163-0.26 (0.03)<0.001-0.11 (0.03)<0.001Men82-0.29 (0.04)<0.001-0.14 (0.05)0.002Women81-0.23 (0.03)<0.001-0.09 (0.03)0.009Adiponectin, mg/Ld1630.04 (0.01)0.0050.01 (0.02)0.78Men820.09 (0.02)<0.0010.04 (0.03)0.11Women81-0.01 (0.02)0.76IGF-1, nmol/L1630.10 (0.24)0.67Men820.36 (0.31)0.24Women81-0.17 (0.35)0.64IGFBP-3, mg/L163-0.05 (0.05)0.37IGF-1:IGFBP-31630.004 (0.003)0.21Men820.009 (0.006)0.14Women81-0.001 (0.003)0.82CRP (high-sensitivity), mg/Ld163-0.11 (0.07)0.09 a P value refers to difference between baseline and end. b P value refers to difference between baseline and end after adjustement for weight loss. c Individuals using antihypertensive agents were removed before analysis. d Natural log transformed parameter was used for analysis. e Individuals using lipid-lowering agents were removed before analysis. Parameters were analysed separately in men and women if there was a significant gender-difference at baseline. BMI, body mass index; REE, resting energy expenditure; FRS, Framingham risk score; HOMA-IR, homeostatic model assessment - insulin resistance; HDL, high density lipoprotein; LDL, low density lipoprotein; fT3, free triiodothyronine; fT4, free thyroxine; TSH, thyroid stimulating hormone; DHEAS, dehydroepiandrosterone-sulfate; IGF-1, insuline-like growth factor 1; IGFBP-3, insulin-like growth factor binding protein 3; CRP, C-reactive protein. Supplementary Table 2B. Effects of the intervention on 1H-NMR metabolites. UnadjustedWeight loss adjustedCharacteristic, mean (SE)nDifferenceP-valueaDifferenceP-valuebAmino acidsAlanine, mmol/L1610.011 (0.070)0.87Men800.011 (0.101)0.92Women810.012 (0.097)0.90Glutamine, mmol/L1620.194 (0.068)0.0050.184 (0.074)0.01Glycine, mmol/L1610.204 (0.042)<0.001Men810.253 (0.058)<0.0010.234 (0.067)0.001Women800.153 (0.061)0.010.113 (0.073)0.12Histidine, mmol/L162-0.524 (0.095)<0.001-0.517 (0.099)<0.001Branched-chain amino acidsIsoleucine, mmol/L161-0.119 (0.062)0.05Men80-0.085 (0.088)0.33Women81-0.152 (0.086)0.08Leucine, mmol/L161-0.178 (0.068)0.008Men80-0.145 (0.089)0.10Women81-0.210 (0.102)0.04-0.121 (0.107)0.26Valine, mmol/L160-0.117 (0.073)0.11Aromatic amino acidsPhenylalanine, mmol/L162-0.015 (0.079)0.85Tyrosine, mmol/L162-0.242 (0.087)0.005-0.163 (0.090)0.07Glycolysis-related metabolitesGlucose, mmol/L162-0.241 (0.069)0.001-0.185 (0.075)0.01Lactate, mmol/L162-0.049 (0.094)0.60Pyruvate, mmol/L161-0.261 (0.086)0.002-0.260 (0.090)0.004Citrate, mmol/L1620.199 (0.078)0.010.220 (0.082)0.007Glycerol, mmol/L157-0.172 (0.068)0.01Men77-0.178 (0.084)0.03-0.107 (0.094)0.25Women80-0.167 (0.106)0.12Ketone bodiesAcetate, mmol/L162-0.012 (0.061)0.85Acetoacetate, mmol/L162-0.097 (0.083)0.24Men81-0.016 (0.122)0.90Women81-0.179 (0.113)0.113-hydroxybutyrate, mmol/L158-0.152 (0.073)0.04-0.089 (0.078)0.26Fatty acidscTotal fatty acids, mmol/L130-0.217 (0.061)<0.001-0.159 (0.069)0.02Omega-3 fatty acids, mmol/L130-0.173 (0.067)0.01-0.154 (0.074)0.04Docosahexaenoic acid (DHA), mmol/L130-0.057 (0.060)0.35Omega-6 fatty acids, mmol/L130-0.322 (0.062)<0.001-0.288 (0.070)<0.001Linoleic acid, mmol/L130-0.262 (0.061)<0.001-0.232 (0.068)0.001Polyunsaturated fatty acids, mmol/L130-0.319 (0.059)<0.001-0.283 (0.067)<0.001Monounsaturated fatty acids, mmol/L130-0.211 (0.057)<0.001-0.114 (0.065)0.08Saturated fatty acids, mmol/L130-0.103 (0.072)0.15Fatty acid chain length1300.414 (0.099)<0.0010.437 (0.104)<0.001Fatty acid ratios, relative to total fatty acidscOmega-3 fatty acids, %130-0.017 (0.085)0.84Docosahexaenoic acid (DHA), %1300.093 (0.073)0.20Omega-6 fatty acids, %130-0.170 (0.073)0.02-0.206 (0.079)0.01Linoleic acid, %130-0.088 (0.060)0.14Polyunsaturated fatty acids, %130-0.166 (0.073)0.02-0.207 (0.080)0.01Monounsaturated fatty acids, %130-0.131 (0.060)0.03-0.027 (0.066)0.68Saturated fatty acids, %1300.378 (0.096)<0.0010.305 (0.100)0.002Unsaturation degree, double bonds per fatty acids130-0.125 (0.078)0.11-0.137 (0.085)0.11Fluid balanceCreatinine, mmol/L162-0.090 (0.050)0.07Men81-0.033 (0.069)0.63Women81-0.146 (0.072)0.04-0.110 (0.078)0.16Albumin, signal area162-0.069 (0.076)0.37InflammationGlycoprotein acetyls, mainly a1-acid glycoprotein, mmol/L162-0.155 (0.058)0.008Men81-0.221 (0.082)0.007-0.126 (0.092)0.16Women81-0.089 (0.083)0.28ApolipoproteinscApolipoprotein B, g/L134-0.311 (0.048)<0.001-0.225 (0.056)<0.001Apolipoprotein A1, g/L134-0.147 (0.052)0.004-0.210 (0.059)<0.001Men65-0.009 (0.071)0.90Women69-0.278 (0.072)<0.001-0.308 (0.081)<0.001LipidscTriglycerides, mmol/L134-0.179 (0.058)0.002Men65-0.290 (0.086)0.001-0.200 (0.097)0.04Women69-0.075 (0.075)0.32Phosphoglycerides, mmol/L130-0.258 (0.068)<0.001-0.276 (0.075)<0.001Men63-0.246 (0.094)0.009-0.323 (0.107)0.003Women67-0.269 (0.097)0.006-0.248 (0.105)0.02Cholines, mmol/L130-0.189 (0.064)0.003-0.218 (0.071)0.002Men63-0.171 (0.090)0.06Women67-0.205 (0.091)0.02-0.183 (0.098)0.06Sphingomyelins, mmol/L130-0.258 (0.051)<0.001-0.286 (0.059)<0.001Men63-0.182 (0.067)0.006-0.281 (0.083)0.001Women67-0.329 (0.076)<0.001-0.313 (0.083)<0.001Total lipid concentrations in lipoprotein subclassescExtremely large VLDL, mmol/L115-0.244 (0.079)0.002-0.235 (0.085)0.006Very large VLDL, mmol/L 103-0.306 (0.084)<0.001-0.277 (0.091)0.002Large VLDL, mmol/L119-0.288 (0.066)<0.001-0.195 (0.072)0.007Medium VLDL, mmol/L 134-0.232 (0.056)<0.001-0.114 (0.063)0.07Small VLDL, mmol/L 134-0.284 (0.055)<0.001-0.176 (0.062)0.005Very small VLDL, mmol/L134-0.283 (0.059)<0.001-0.239 (0.067)<0.001IDL, mmol/L 134-0.245 (0.051)<0.001-0.222 (0.059)<0.001Large LDL, mmol/L 134-0.232 (0.052)<0.001-0.188 (0.060)0.002Medium LDL, mmol/L 134-0.240 (0.055)<0.001-0.179 (0.063)0.004Small LDL, mmol/L 134-0.266 (0.054)<0.001-0.205 (0.061)0.001Very large HDL, mmol/L 1330.069 (0.061)0.26Men640.217 (0.094)0.020.025 (0.096)0.80Women69-0.068 (0.075)0.36Large HDL, mmol/L1260.091 (0.046)0.05Men590.241 (0.079)0.0020.077 (0.087)0.38Women67-0.042 (0.048)0.38Medium HDL, mmol/L 134-0.115 (0.068)0.09Men650.025 (0.091)0.79Women69-0.247 (0.098)0.01-0.246 (0.105)0.02Small HDL, mmol/L 134-0.200 (0.088)0.02-0.111 (0.092)0.228Men65-0.215 (0.120)0.07Women69-0.185 (0.129)0.15Lipoprotein particle sizecVLDL diameter, nm134-0.136 (0.067)0.04-0.032 (0.073)0.66LDL diameter, nm1340.115 (0.095)0.23HDL diameter, nm1340.108 (0.050)0.03-0.051 (0.054)0.35Men650.258 (0.077)0.0010.081 (0.081)0.32Women69-0.032 (0.060)0.59CholesterolcTotal cholesterol, mmol/L134-0.296 (0.051)<0.001-0.279 (0.059)<0.001VLDL cholesterol, mmol/L134-0.301 (0.059)<0.001-0.223 (0.067)0.001IDL cholesterol, mmol/L134-0.265 (0.051)<0.001-0.249 (0.059)<0.001LDL cholesterol, mmol/L134-0.246 (0.054)<0.001-0.197 (0.061)0.001HDL cholesterol, mmol/L1340.005 (0.043)0.90Men650.140 (0.062)0.020.015 (0.072)0.83Women69-0.122 (0.055)0.03-0.174 (0.066)0.008HDL2 cholesterol, mmol/L1340.023 (0.042)0.58Men650.163 (0.061)0.0080.036 (0.073)0.62Women69-0.108 (0.055)0.05-0.160 (0.066)0.02HDL3 cholesterol, mmol/L134-0.023 (0.074)0.75Men65-0.005 (0.104)0.96Women69-0.041 (0.107)0.70 a P value refers to difference between baseline and end. b P value refers to difference between baseline and end after adjustment for weight loss. c Individuals using lipid-lowering agents were removed before analysis. All metabolite concentrations were natural log-transformed and scaled to standard deviation units before analysis. Parameters were analysed separately in men and women if there was a significant gender-difference at baseline. VLDL, very low density lipoprotein; IDL, intermediate density lipoprotein; LDL, low density lipoprotein; HDL, high density lipoprotein. Suplementary Figures  Supplementary Figure 1. Effect of the intervention on insulin levels.  Supplementary Figure 2. Association of 1H-NMR metabolites with PCs identified using PCA analysis. The colour of the cells represents the magnitude of the effect.     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lBapPytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V lJ t6,5&5+5555559/ BpPytLkkdF$$IflJִT?<q$)16e&e+eeeeee t6    44 lBapPytLk$$If!vh#v#v#v#v5#v&#v,#v/:V lJ t6,5R5555559/ BpFytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V l t6,5&5+5555559/ BpPytLkkdJ$$IflִT?<q$)16e&e+eeeeee t6    44 lBapPytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V lw t6,5&5+5555559/ BpPytLkkdM$$IflwִT?<q$)16e&e+eeeeee t6    44 lBapPytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V lJ t6,5&5+5555559/ BpPytLkkdP$$IflJִT?<q$)16e&e+eeeeee t6    44 lBapPytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V l; t6,5&5+5555559/ BpPytLkkdS$$Ifl;ִT?<q$)16e&e+eeeeee t6    44 lBapPytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V l; t6,5&5+5555559/ BpPytLkkdV$$Ifl;ִT?<q$)16e&e+eeeeee t6    44 lBapPytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V l; t6,5&5+5555559/ BpPytLkkdY$$Ifl;ִT?<q$)16e&e+eeeeee t6    44 lBapPytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V l; t6,5&5+5555559/ BpPytLkkd\$$Ifl;ִT?<q$)16e&e+eeeeee t6    44 lBapPytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V l; t6,5&5+5555559/ BpPytLkkd_$$Ifl;ִT?<q$)16e&e+eeeeee t6    44 lBapPytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V lJ t6,5&5+5555559/ BpPytLkkdb$$IflJִT?<q$)16e&e+eeeeee t6    44 lBapPytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V l; t6,5&5+5555559/ BpPytLkkde$$Ifl;ִT?<q$)16e&e+eeeeee t6    44 lBapPytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V l; t6,5&5+5555559/ BpPytLkkdh$$Ifl;ִT?<q$)16e&e+eeeeee t6    44 lBapPytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V l; t6,5&5+5555559/ BpPytLkkdk$$Ifl;ִT?<q$)16e&e+eeeeee t6    44 lBapPytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V l; t6,5&5+5555559/ BpPytLkkdn$$Ifl;ִT?<q$)16e&e+eeeeee t6    44 lBapPytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V l; t6,5&5+5555559/ BpPytLkkdq$$Ifl;ִT?<q$)16e&e+eeeeee t6    44 lBapPytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V l; t6,5&5+5555559/ BpPytLkkdt$$Ifl;ִT?<q$)16e&e+eeeeee t6    44 lBapPytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V l; t6,5&5+5555559/ BpPytLkkdw$$Ifl;ִT?<q$)16e&e+eeeeee t6    44 lBapPytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V l; t6,5&5+5555559/ BpPytLkkdz$$Ifl;ִT?<q$)16e&e+eeeeee t6    44 lBapPytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V l; t6,5&5+5555559/ BpPytLkkd}$$Ifl;ִT?<q$)16e&e+eeeeee t6    44 lBapPytLk$$If!vh#v#v#v#v5#v&#v,#v/:V l; t6,5R5555559/ BpFytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V l; t6,5&5+5555559/ BpPytLkkd$$Ifl;ִT?<q$)16e&e+eeeeee t6    44 lBapPytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V lw t6,5&5+5555559/ BpPytLkkd$$IflwִT?<q$)16e&e+eeeeee t6    44 lBapPytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V lJ t6,5&5+5555559/ BpPytLkkd$$IflJִT?<q$)16e&e+eeeeee t6    44 lBapPytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V lw t6,5&5+5555559/ BpPytLkkd$$IflwִT?<q$)16e&e+eeeeee t6    44 lBapPytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V lw t6,5&5+5555559/ BpPytLkkd$$IflwִT?<q$)16e&e+eeeeee t6    44 lBapPytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V l; t6,5&5+5555559/ BpPytLkkd$$Ifl;ִT?<q$)16e&e+eeeeee t6    44 lBapPytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V l; t6,5&5+5555559/ BpPytLkkd$$Ifl;ִT?<q$)16e&e+eeeeee t6    44 lBapPytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V lw t6,5&5+5555559/ BpPytLkkd$$IflwִT?<q$)16e&e+eeeeee t6    44 lBapPytLk$$If!vh#vA#v#v#v#v5#v&#v,#v/:V lw t6,5&5+5555559/ 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