Bilberry supplementation in type 2 diabetes

INTERVENTION: Individuals who meet all inclusion criteria and none of the exclusion criteria will, after providing both oral and written informed consent to participate in the project, be randomised according to a computer generated random sequence. Block randomisation will be performed in a 1:1 ratio to the respective intervention. Both study participants and research staff involved in the project will be blinded to treatment allocation. Arm 1: Dietary Intervention: Bilberry beverage. Participants randomized to this group will be instructed to daily consume 500 ml of bilberry beverage for 12 weeks (± 7 days). The beverage contains naturally occurring bioactive compounds derived from bilberries, including polyphenols such as anthocyanins. Both the bilberry and control beverages are isocaloric and matched for taste, appearance, and mouth feel. Participants are instructed to consume the beverage daily as part of their usual routines, with the total daily intake divided into at least two doses (250 ml in the morning and 250 ml in the evening). Compliance will be assessed using two complementary methods. Participants will be asked to return any unopened beverages at the follow‐up visit, with the number of unopened cartons subtracted from the total amount provided to estimate actual intake. Compliance will also be evaluated objectively by quantifying anthocyanin‐derived metabolites in faecal samples collected at follow‐up Arm 2: Placebo comparator: Control beverage. Participants randomized to this group will be instructed to daily consume 500 ml of the control beverage for 12 weeks (± 7 days). The control beverage does not contain bilberry‐derived bioactive compounds but is isocaloric and matched to the bilberry beverage for taste, appearance, and mouth feel. Participants are instructed to consume the beverage daily as part of their usual routines, with the total daily intake divided into at least two doses (250 ml in the morning and 250 ml in the evening). Compliance will be assessed by asking participants to return any unopened beverages at the follow‐up visit, with the number of unopened cartons subtracted from the total number provided to estimate actual intake. CONDITION: Cardiometabolic health, exercise capacity and inflammation in individuals with type 2 diabetes ; Nutritional, Metabolic, Endocrine PRIMARY OUTCOME: Oxidized low density lipoprotein (oxLDL) measured using Mercodia Oxidized LDL ELISA kit reported in U/L at baseline and after 12 weeks of intervention (±7 days) SECONDARY OUTCOME: 8‐oxo‐2'‐deoxyguanosine (8‐oxo‐dG) measured using morning urine and reported in ng/ml or nmol/l at baseline and after 12 weeks of intervention (±7 days) Alanine aminotransferase (ALAT) measured using serum and reported in U/L at baseline and after 12 weeks of intervention (±7 days) Alkaline phosphatase (BASP) measured using serum and reported in U/L at baseline and after 12 weeks of intervention (±7 days) Apolipoprotein A‐I (ApoA) measured using serum or plasma after an overnight fast and reported in g/L at baseline and after 12 weeks of intervention (±7 days) Apolipoprotein B (ApoB) measured using serum and reported in g/L at baseline and after 12 weeks of intervention (±7 days) Aspartate aminotransferase measured using serum and reported in U/L at baseline and after 12 weeks of intervention (±7 days) Bilirubin measured using serum and reported in µmol/L at baseline and after 12 weeks of intervention (±7 days) Blood pressure (systolic and diastolic) measured using a seated position after a resting period of minimum 15 minutes and reported in mmHg at baseline and after 12 weeks of intervention (±7 days) Body composition measured using bioelectrical impedance analysis (BIA) at baseline and after 12 weeks of intervention (±7 days) Body composition measured using bioelectrical impedance analysis and reported as body composition parameters including fat mass, lean mass, and body fat percentage at baseline and after 12 weeks of intervention (±7 days) Calcium measured using serum or plasma and reported in mmol/l at baseline and after 12 weeks of intervention (±7 days) Circulating metabolites (plasma metabolomics) measured using metabolomic profiling of plasma samples and reported as relative or absolute metabolite concentrations depending on analytical platform at baseline and after 12 weeks of intervention (±7 days) Coagulation factors II, VII, and Xactivity measured using combined activity of coagulation factors II, VII, and Xmeasured in plasma and reported in arbitrary units or percentage depending on assay method at baseline and after 12 weeks of intervention (±7 days) Creatinine measured using serum or plasma and reported in µmol/L at baseline and after 12 weeks of intervention (±7 days) Cystatin C measured using serum or plasma and reported in mg/L at baseline and after 12 weeks of intervention (±7 days) Endothelial activation measured using plasma concentrations of syndecan‐1 (unit of measure: ng/ml) and thrombomodulin (unit of measure: ng/ml) measured with commercial ELISA kits (CD138 kit and CD141 kit, Diaclone, Medi XBiochemica, Espoo, Finland) at baseline and after 12 weeks of intervention (±7 days) Erythrocyte mean cell volume (MCV) measured using whole blood and reported in fL at baseline and after 12 weeks of intervention (±7 days) Estimated glomerular filtration rate (eGFR) measured using serum creatinine and/or cystatin C and reported in ml/min/1.73 m² at baseline and after 12 weeks of intervention (±7 days) Fasting C‐peptide measured using plasma after an overnight fast and reported in pmol/L at baseline and after 12 weeks of intervention (±7 days) Fasting insulin measured using plasma after an overnight fast and reported in pmol/L at baseline and after 12 weeks of intervention (±7 days) Fasting plasma glucose measured using plasma after an overnight fast and reported in mmol/L at baseline and after 12 weeks of intervention (±7 days) Fibrinolysis measured using e Xvivo fibrinolytic capacity (fibrinolysis speed) in whole blood measured with in‐house modified rotational thromboelastometry after stimulation with tissue factor and tissue plasminogen activator. Unit of measure: mm/min. E Xvivo fibrinolytic capacity (time from peak fibrin to 50% lysis) in platelet‐poor plasma measured with in‐house turbidimetric fibrin formation and lysis assay. Unit of measure: seconds. Plasma concentrations of plasminogen activity (unit of measure: IU/L), tissue plasminogen activator (unit of measure: ng/mL) and plasminogen activator inhibitor‐1 (unit of measure: ng/mL) measured with ELISA (Technozym®, Technoclone, Vienna, Austria) at baseline and after 12 weeks of intervention (±7 days) Glycaemic variability measured using continous glucose montitoring (CGM) at 14 days before baseline and 14 days before the end of 12 weeks of intervention (±7 days) Glycated haemoglobin A1c (HbA1c) measured using whole blood and reported in mmol/mol at baseline and after 12 weeks of intervention (±7 days) Gut microbiota composition measured using microbial profiling methods on faecal samples at baseline and after 12 weeks of intervention (±7 days) Haemoglobin measured using concentration in whole blood and reported in mmol/l at baseline and after 12 weeks of intervention (±7 days) Handgrip strength measured using a hand dynamometer and reported in kg at baseline and after 12 weeks of intervention (±7 days) Hemostatic function: Primary hemostasis measured using e Xvivo platelet aggregation in whole blood measured with impedance aggregometry (Multiplate®, Roche, Switzerland) after stimulation with adenosine diphosphate (ADP), arachidonic acid and thrombin‐related activation peptide (TRAP‐6). Reported in aggregation units Xmin. Plasma concentration of P‐selectin measured with commercial ELISA (CD62P Quantikine, Biotechne, Dublin, Ireland) and reported in ng/ml at baseline and after 12 weeks of intervention (±7 days) High‐density lipoprotein (HDL)‐cholesterol measured using serum and reported in mmol/L at baseline and after 12 weeks of intervention (±7 days) High‐sensitivity C‐reactive protein (hs‐CRP) measured using serum or plasma and reported in mg/L at baseline and after 12 weeks of intervention (±7 days) Homeostatic Model Assessment (HOMA) measured using fasting glucose and C‐peptide at baseline and after 12 weeks of intervention (±7 days) Immature platelet fraction (IPF) measured using whole blood and reported as percentage of total platelet count (%) at baseline and after 12 weeks of intervention (±7 days) Interleukin‐6 (IL‐6) measured using Olink 96 Target Inflammation panel and reported in NP X(normalized protein expression) at baseline and after 12 weeks of intervention (±7 days) International normalised ratio (INR) measured using plasma based on coagulation factors II, VII, and Xactivity and reported as a unitless ratio at baseline and after 12 weeks of intervention (±7 days) Lactate dehydrogenase (LDH) measured using serum and reported in U/L at baseline and after 12 weeks of intervention (±7 days) Leukocyte count measured using whole blood and reported in ×10?/l at baseline and after 12 weeks of intervention (±7 days) Lipoprotein(a) [Lp(a)] measured using serum or plasma after an overnight fast and reported in nmol/L at baseline and after 12 weeks of intervention (±7 days) Low‐density lipoprotein (LDL)‐cholesterol measured using the Friedewald equation from total cholesterol, HDL‐cholesterol and triglycerides and reported in mmol/L at baseline and after 12 weeks of intervention (±7 days) Markers of inflammation measured using O‐link Target‐96 Inflammation panel at baseline and after 12 weeks of intervention (±7 days) Peripheral neuropathy measured using iothesiometry to assess vibration perception threshold and reported in volts at baseline and after 12 weeks of intervention (±7 days) Phosphate measured using serum or plasma and reported in mmol/l at baseline and after 12 weeks of intervention (±7 days) Physical function measured using 6‐minute walk test (total walking distance in metres over 6 minutes on a standardised course) at baseline and after 12 weeks of intervention (±7 days) Platelet count measured using whole blood and reported in ×10?/l at baseline and after 12 weeks of intervention (±7 days) Postprandial glucose response during a mixed meal test measured using continuous glucose monitoring (CGM) at baseline and after 12 weeks of intervention (±7 days) Potassium measured using serum or plasma and reported in mmol/l at baseline and after 12 weeks of intervention (±7 days) Retinal changes measured using retinal examination of the ocular fundus using standard ophthalmological assessment methods at baseline and after 12 weeks of intervention (±7 days) Secondary hemostasis measured using e Xvivo thrombin generation (endogenous thrombin potential) in platelet‐poor plasma measured using Calibrated Automated Thrombogram (BV Thrombinoscope, Maastricht, the Netherlands) after stimulation with tissue factor (reported in nM Xmin). Plasma concentration of prothrombin fragment 1+2, measured with commercial ELISA (EnzygnostTM, Siemens Healthineers, Ballerup, Denmark) and reported in pmol/L at baseline and after 12 weeks of intervention (±7 days) Self‐reported physical activity level measured using the Frändin/Grimby activity scale questionnaire and reported as categorical activity level scores based on self‐reported habitual physical activity at baseline and after 12 weeks of intervention (±7 days) Sit‐to‐stand test measured using time required to complete five consecutive sit‐to‐stand repetitions from a chair without use of the arms and reported in seconds at baseline and after 12 weeks of intervention (±7 days) Small dense low‐density lipoprotein (sdLDL)‐cholesterol measured using using ELISA (Bio‐Techne/NOVUS NBP2‐82535 kit) on serum or plasma after an overnight fast and reported in nmol/ml at baseline and after 12 weeks of intervention (±7 days) Sodium measured using serum or plasma and reported in mmol/l at baseline and after 12 weeks of intervention (±7 days) Three‐day food self‐reported diary measured using the myfood24 dietary assessment platform at baseline and after 12 weeks of intervention (±7 days) Triglycerides measured using serum after an overnight fast and reported in mmol/L at baseline and after 12 weeks of intervention (±7 days) U‐Albumin/creatinine‐ratio measured using morning urine samples and reported in mg/g at baseline and after 12 weeks of intervention (±7 days) INCLUSION CRITERIA: 1. Diagnosed type 2 diabetes mellitus 2. Age =18 years 3. Written informed consent for participation