Emerging evidence suggests that following a 'carbohydrate-last meal pattern', wherein foods rich in protein, fat, fiber, and/or polyphenols are consumed before sources of simple carbohydrate (CHO) in a meal, results in reduced postprandial glycaemic responses than the reverse food order or a co-ingestion pattern. This effect has been observed across the spectrum of glucose tolerance, from patients with diabetes to individuals with normal glucose tolerance (Kuwata et al., 2016; Nishino et al., 2018; Lu et al., 2019; Sun et al., 2020). Furthermore, reduced glucose excursions have been linked to decreased subsequent hunger and energy intake (Lu et al., 2019; Wyatt et al., 2021). However, to date, no studies on food intake sequence have targeted athletes, despite their increased CHO demands (Thomas et al., 2016) which could expose them to repeated episodes of hyperglycaemia and high glycaemic variability, known to increase the risk of adverse cardiovascular outcomes and all-cause mortality (Loader et al., 2015; Cavero-Redondo et al., 2017; Faerch et al., 2018). Additionally, athletes often face pressure to meet body composition standards and may benefit from strategies that enhance satiety and craving control. Finally, there is reason to believe that better glycaemic control could lead to improved performance, given that enhancements in endurance activities have been observed with a low-glycemic-index diet compared to a high-glycemic-index diet (Heung-Sang Wong et al., 2017). Therefore, this randomised crossover trial is part of a wider project which seeks to explore the impact of food intake sequence on metabolic health and performance in athletes. Specifically, this trial aims to investigate the acute, postprandial metabolic and appetite responses to consuming an identical meal in two intake sequences (CHO-last versus CHO-first) in athletes, while in the resting state.
Participants will be required to visit the research facilities at Cidade do Futebol (Portugal Football School's headquarters) on three separate occasions, following a 10-12-hour overnight fast and abstaining from alcohol consumption and strenuous physical activity the day before (e.g. no exercise causing sweating or heavy breathing). Visits will be separated by a wash-out period of 7 days to ensure participants experience similar dietary and physical activity patterns in the 24 hours preceding the trials. Each visit is expected to last approximately 4 hours. In the screening and familiarisation visit, athletes will be asked to provide written informed consent to participate in the study and answer a series of questions to confirm their eligibility and safety for enrolment. Subsequently, an anthropometric assessment, blood pressure measurement, fasted blood collection and 2-hour 75-g oral glucose tolerance test (OGTT) will be performed to establish participants' baseline characteristics and analyse biochemical markers of the glucose and lipid metabolism to identify any further exclusion criteria (i.e., glycated haemoglobin (HbA1c); glucose; insulin; triglycerides; total, high-density lipoprotein and low-density lipoprotein cholesterol; haemogram and high-sensitivity C-reactive protein). To familiarise participants with the cannulation procedure ahead of the experimental trials, a trained and experienced nurse will insert a cannula (a small plastic tube) into a vein on the participants' arm, from which fasting and 2-h OGTT blood samples will be drawn (15 ml in total). Eligible participants will be instructed to monitor their diet and physical activity for 24 hours before their first experimental visit and replicate these patterns before the second experimental visit. In both experimental visits, participants will arrive at the research facilities between 8:00-9:00 and rest comfortably for 10 minutes. Body mass and blood pressure will be remeasured. Then, cannulation will be performed, and 18.5 ml venous blood samples will be drawn immediately before and at 30, 60, 90, 120, and 180 minutes after the meal challenge to assess postprandial changes in insulin, glucagon-like peptide-1 (GLP-1), glucose-dependent insulinotropic polypeptide (GIP), ghrelin, peptide YY (PYY), triglycerides, and non-esterified fatty acids (NEFA) concentrations. At similar timepoints, as well as at 15 and 45 minutes, capillary blood glucose will be measured using a finger prick glucometer, and appetite ratings will be marked by the athletes on 100-mm visual analogue scales. Upon completing the research assessments, participants will be asked to photograph and record the timing, type, and amounts of foods, drinks, and/or supplements consumed for an additional 3 hours to evaluate their prospective, ad libitum intake.
Study Type
INTERVENTIONAL
Allocation
RANDOMIZED
Purpose
BASIC_SCIENCE
Masking
SINGLE
Enrollment
14
Skyr yoghurt, whey protein and almonds over \~5 min, immediately followed by white bread, strawberry jam, banana and pulp-free orange juice over \~10 min.
White bread, strawberry jam, banana and pulp-free orange juice over \~10 min, immediately followed by skyr yoghurt, whey protein and almonds over \~5 min.
Cidade do Futebol, Avenida das Seleções
Oeiras, Lisbon District, Portugal
Postprandial blood glucose concentrations
Difference between food intake sequences in the incremental area under the curve 0-120 and 0-180 min; peak concentrations (mmol/L); time-to-peak (min); and change from baseline data (mmol/L) of capillary blood glucose measured by a validated glucometer
Time frame: Baseline and 15, 30, 45, 60, 90, 120, 180 minutes following the test meal
Postprandial subjective appetite ratings
Difference between food intake sequences in the total area under the curve 0-180 min; peak/nadir rating (mm); time-to-peak/nadir (min); and change from baseline data (mm) of appetite ratings measured by validated 100-mm visual analogue scales (Flint et al, 2000)
Time frame: Baseline and 15, 30, 45, 60, 90, 120, 180 minutes following the test meal
Prospective ad libitum energy intake
Difference between food intake sequences in the total energy (kcal) consumed within 3 hours from the end of the experimental visit, assessed by a photography-supported dietary record of foods and fluids
Time frame: 0-3 hours post-trial
Prospective ad libitum nutritional intake
Difference between food intake sequences in the total carbohydrate, protein and fat (grams; g/kg of body mass and % of total energy intake) consumed within 3 hours from the end of the experimental visit, assessed by a photography-supported dietary record of foods and fluids
Time frame: 0-3 hours post-trial
Postprandial serum insulin concentrations
Difference between food intake sequences in the incremental area under the curve 0-120 and 0-180 min; peak concentrations (microIU/ml); time-to-peak (min); and change from baseline data (microIU/ml) of serum insulin measured by a chemiluminescent immunoassay
Time frame: Baseline and 30, 60, 90, 120, 180 minutes following the test meal
Postprandial plasma total GLP-1 concentrations
Difference between food intake sequences in the incremental area under the curve 0-120 and 0-180 min; peak concentrations (pg/ml); time-to-peak (min); and change from baseline data (pg/ml) of plasma GLP-1 measured by a total GLP-1 ELISA kit
Time frame: Baseline and 30, 60, 90, 120, 180 minutes following the test meal
Postprandial plasma total GIP concentrations
Difference between food intake sequences in the incremental area under the curve 0-120 and 0-180 min; peak concentrations (pg/ml); time-to-peak (min); and change from baseline data (pg/ml) of plasma GIP measured by a total GIP ELISA kit
Time frame: Baseline and 30, 60, 90, 120, 180 minutes following the test meal
Postprandial serum total ghrelin concentrations
Difference between food intake sequences in the total area under the curve 0-120 and 0-180 min; nadir concentrations (pg/ml); time-to-nadir (min); and change from baseline data (pg/ml) of serum ghrelin measured by a total ghrelin ELISA kit
Time frame: Baseline and 30, 60, 90, 120, 180 minutes following the test meal
Postprandial plasma total PYY concentrations
Difference between food intake sequences in the incremental area under the curve 0-120 and 0-180 min; peak concentrations (pg/ml); time-to-peak (min); and change from baseline data (pg/ml) of plasma PYY measured by a total PYY ELISA kit
Time frame: Baseline and 30, 60, 90, 120, 180 minutes following the test meal
Postprandial serum triglyceride concentrations
Difference between food intake sequences in the incremental area under the curve 0-180 min; peak concentrations (mmol/L); time-to-peak (min); and change from baseline data (mmol/L) of serum triglycerides measured by the GPO/POD method
Time frame: Baseline and 30, 60, 90, 120, 180 minutes following the test meal
Postprandial serum NEFA concentrations
Difference between food intake sequences in the total area under the curve 0-180 min; nadir concentrations (mmol/L); time-to-nadir (min); and change from baseline data (mmol/L) of serum NEFA measured by an enzymatic colorimetric assay
Time frame: Baseline and 30, 60, 90, 120, 180 minutes following the test meal
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