Appetite assessments
appetite sensations
Participants attended the clinic following an overnight fast (> 10 h). Baseline blood samples were taken using an antecubital vein catheter, after which participants consumed their randomly allocated snack (almonds or a carbohydrate-based snack) with 200 mls of water within a 10-min period. Repeat blood sampling (via canulation) was performed every 30 min post snack for 2 h. Water (200 mls) was given at 60 min, and 100 mls at 90 min after snack consumption. Immediately after each blood draw, protease inhibitors (Sigma P2714 and Millipore DPP4-010) were added to the specimen tube. All samples were stored as serum at − 80 °C until assayed in duplicate. Appetite hormones ghrelin, GIP, GLP-1, leptin, PP, PYY as well as C-peptide and glucagon were assessed using a multiplex analysis system (LUMINEX MAGPIX, Millipore, Merck). CCK was assessed using ELISA (Ray Biotech). All samples for the same participant were run in the same assay.Participants were asked to rate their subjective appetite sensations by answering four questions at the time of each blood draw. Each question was answered using a visual analogue scale (VAS); a 10 cm horizontal line anchored at both ends so that answers could be indicated on a continuous scale. VAS for appetite assessment has been shown to have good validity, reliability, and reproducibility [