Please add notes for methods/analysis regarding the VDP. Add contact name for potential questions) ### METHODS FOR SAMPLES COLLECTION/ HANDELING/ BLOOD ANALYSES A home sampling kit was provided to all participants which included detailed picture illustrated instructions for sampling, 3 barcoded tubes for stool samples, a flushable sheet, spoons and an opaque zip-lock bag. Participants were instructed to lay the sheet in a tray to collect their fresh fecal sample, enabling sampling without contact with urine or toilet water. After that, participants were told to transfer feces from the tray into the tubes using the spoons provided, right after completing toileting. Closed tubes were stored in the opaque bags and immediately stored in home freezers (-18°C). together with accurate medical information, extensive lifestyle and early life events data and blood values were combined in a geographically confined region. In short, people in Flanders (+/- 6.5 million inhabitants, +/- 13500 km2) could subscribe online to participate in this project from January 2013 onwards. Sampling rounds were organized between June 2013 and January 2014. At their subscription, they got information about the study and could consent for receiving a sampling kit. They also filled in a first questionnaire about early life events and general information. When a sampling round was performed, participants received a sampling kit about one month before. They were instructed to make an appointment with their general practitioner, to go there fasted for blood sampling, medical check-up and filling in an extended medical questionnaire including medication usage in the past 6 months and antibiotic usage in the past 12 months. After the GP visit, they used the sampling kit we had provided to collect and store their feces and saliva at home. Samples were fresh frozen in household freezers and transported frozen within a week to one of the collection points. Twice a week, the frozen samples were picked up at the collection points and transported on dry ice to the lab. Blood samples were analyses were performed at ### STOOL SAMPLE EXTRACTION PROTOCOL ### SAMPLE SELECTION Freeze1: N=1198 1205 samples had no inconsistencies between medical (GP) and blood (CMA) data (gender, birthdate), and with quality checks on BMI extremes and random selection of samples. Under-age (<18) individuals were excluded. 5 samples were subsequently excluded due to low sequencing depth. 2 samples were subsequently excluded due to incongruence between medical/CMA and database gender and birthdate. ### 16S DATA FILES (Jun) localization (zeus): /raeslab/group/data/MiSeq_data_V4/FINAL1200 ### METADATA FILES # VDPMAIN_METADATA contains: 1) ids (sample,database,CMA), Age, Gender, BMI, blood pressure (BP.sys,BP.dia) (GP origin) and smoking + basic (study level, marital status, ethnicity, geoloc) info (self reported) #QC done (Sara) 2) All clinical (blood) variables (CMA) #QC done (Sara) 3) Health scores (Sara) 4) enterotyping (original JSD (3ETs), JSD (2ETs) and DMM (4ETs)) (Doris) Note: Smoker (1=defined has having smoked during the past 3 months) Summary of main fields of interest: Gender F M Count 652.00 546.00 Percent 54.42 45.58 Smoker 0 1 <NA> Count 1032.00 106.00 60.00 Percent 86.14 8.85 5.01 Age BMI height weight Sedimentation.1h.mm Hemoglobine.g.dL Hematocriet..perc RBC.milj.mm3 MCV.fl MCH.pg MCHC.g.dL WBC..mm3 Lymfocyten..perc Lymfocyten..mm3 Neutrofiele.segmenten..perc Eosinofielen.perc Monocyten.perc Monocyten..mm3 Basofielen..perc Basofielen..mm3 Neutrofielen..mm3 Eosinofielen..mm3 Trombocyten.1000.mm3 Vit..B12.ng.L CRP.mg.L HbA1c.mmol.mol HbA1c..perc Gluc.nuchter.mg.dL Insuline.nuchter.mU.L Ureum.mg.dL Creatinine.mg.dL e.GFR..CKD.EFI..mL.min.1.73m2 Urinezuur.mg.dL Ferritine.microg.L Totaal.Eiwit.g.L Albumine..perc Albumine.g.L Alfa1Globulines..perc Alfa1Globulines.g.L Alfa2Globulines..perc Alfa2Globulines.g.L BetaGlobulines..perc BetaGlobulines.g.L GammaGlobulines..perc GammaGlobulines.g.L mean sd median min max valid.n 50.88 13.78 53.00 18.00 85.00 1198 24.91 4.35 24.14 16.18 51.89 1198 171.12 8.64 170.00 149.00 198.00 1198 73.17 15.07 71.00 43.00 143.00 1198 6.26 7.02 5.00 1.00 116.00 1195 14.29 1.20 14.30 10.10 18.10 1197 42.45 3.27 42.40 29.00 52.90 1197 4.68 0.39 4.67 3.17 6.21 1197 90.87 4.64 91.00 61.00 109.00 1197 30.61 1.61 31.00 19.00 37.00 1197 33.74 1.21 34.00 28.00 38.00 1197 5974.98 1619.81 5750.00 2250.00 17260.00 1197 33.31 7.95 33.30 8.50 65.00 1197 1943.50 566.34 1881.00 540.00 6149.00 1197 54.44 8.69 54.40 30.50 82.80 1197 2.90 1.96 2.40 0.00 13.80 1197 8.80 2.21 8.60 0.00 20.80 1197 518.08 170.45 492.00 0.00 1774.00 1197 0.55 0.35 0.50 0.00 3.20 1197 31.23 19.15 29.00 0.00 169.00 1197 3313.67 1270.10 3063.00 1131.00 11564.00 1197 168.54 121.03 139.00 0.00 1318.00 1197 247.71 57.45 244.00 82.00 753.00 1197 510.65 279.17 458.00 30.00 2000.00 1198 2.01 4.01 0.90 0.30 64.10 1198 37.31 5.23 37.00 15.00 89.00 1198 5.56 0.48 5.50 3.50 10.30 1198 87.98 16.92 85.00 28.00 309.00 1198 9.03 8.88 7.10 0.40 164.90 1198 30.35 8.06 29.60 9.10 63.50 1198 0.84 0.17 0.83 0.44 1.91 1198 84.72 8.78 90.00 35.00 90.00 1095 4.88 1.25 4.80 2.20 10.50 1198 145.20 124.01 118.00 5.00 1088.00 1198 69.92 3.94 70.00 55.00 86.00 1198 61.98 3.08 62.20 49.60 74.70 1198 43.31 2.91 43.30 29.20 56.00 1198 3.75 0.61 3.70 2.10 8.80 1198 2.61 0.41 2.60 1.60 4.80 1198 9.13 1.34 9.00 4.90 17.60 1198 6.38 0.93 6.30 3.40 11.10 1198 10.89 1.35 10.80 6.30 18.30 1198 7.62 1.06 7.50 3.80 13.40 1198 14.25 2.36 14.20 1.20 24.00 1198 10.00 1.96 9.90 0.70 18.50 1198 Chol..Totaal.mg.dL Triglyceriden.mg.dL HDL.chol..mg.dL Ratio.chol..tot..HDL.chol.mg.mg LDL.chol..gem..mg.dL Bilirubine.tot..mg.dL Bilirubine.dir..mg.dL Bilirubine.indir..mg.dL GOT.U.L GPT.U.L Gamma.GT.U.L CK.U.L Cortisol.ochtend.mcg.dL BP.sys BP.dia SCORE GFR_CDK.EPI PhysioHealthScore Cardiov_penalty Nefrol_penalty 197.87 101.55 64.44 3.37 118.13 0.61 0.22 0.39 21.81 22.66 25.06 109.85 16.03 122.98 76.43 2.09 91.88 97.10 0.04 0.06 37.78 62.70 17.24 4.10 33.44 0.31 0.09 0.23 8.97 13.28 25.17 68.91 7.44 14.08 8.32 2.81 16.03 3.23 0.06 0.10 196.00 86.00 62.00 3.09 116.00 0.54 0.20 0.34 20.00 19.00 17.00 92.00 15.20 120.00 80.00 1.00 92.08 98.31 0.02 0.00 83.00 20.00 23.00 1.36 32.00 0.11 0.10 0.10 7.00 6.00 3.00 15.00 0.50 80.00 40.00 0.00 35.46 77.64 0.00 0.00 472.00 683.00 165.00 141.00 376.00 2.68 0.76 1.99 135.00 122.00 232.00 711.00 49.30 190.00 108.00 18.00 141.70 100.00 0.38 0.61 1198 1198 1198 1198 1198 1197 1197 1197 1198 1198 1198 1198 1198 1109 1104 1053 1198 1051 1053 1198 # VDP metadata numeric contains: 1) vdpmetadata_countscontinuous.txt: continuous metadata in VDPMAIN_METADATA with dates converted into numeric values (Karoline) 2) vdpmetadata_binary.txt: binary and binarized categorical metadata in VDPMAIN_METADATA (Karoline) 3) README_continuous.txt: Details on 1) (Karoline) 4) README_binary.txt Details on 2) (Karoline) localization (owncloud): VDP feeze 1/VDP metadata numeric