The novel supplement group consumed two portions of the novel supplement (Vital01, VitalNext B.V., Wageningen, The Netherlands) per day, each portion consisting of 63 grams of powder to be dissolved in a liquid of choice. The standard supplement group consumed two 200 ml bottles of Nutridrink (Nutridrink, Nutricia Advanced Medical Nutrition, Danone, Hoofddorp, The Netherlands) per day. The daily doses of the novel and the standard supplement product contained equal amounts of energy, carbohydrates, fats and protein, but differed in the type of protein and the amount of BCAAs, vitamin D and UA (see Table 1 for the nutritional content). Participants were advised to consume products after breakfast and lunch, but deviation from this advice was allowed to maximize compliance. On measurement days, participants were not allowed to consume products prior to the university visit.
Nutridrink
Manufactured by Nutricia
Sourced in Netherlands, Italy
Nutridrink is a milk-based nutritional drink designed to provide balanced nutrition. It contains a combination of proteins, carbohydrates, and essential vitamins and minerals. Nutridrink is intended to be a dietary supplement.
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25 protocols using nutridrink
1
Comparison of Novel and Standard Supplements
2
Standard Liquid Meal Composition
3
Detailed Metabolic Monitoring Protocol
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Participants were sampled in the same research room and received standardized feeding at three fixed times during the day (between 09.00 h–10.00 h, 12.00 h–13.00 h and 18.00–19.00 h), each consisting of 600 kcal Nutridrink (Nutricia Advanced Medical Nutrition, Zoetermeer, The Netherlands). No naps were allowed during the day and lights were turned off between 23.00 h to 08.00 h. A catheter was placed in a vein of the forearm of the non-dominant hand. Every 10 minutes, 1.2 ml of blood was collected in K3-EDTA tube and 2 ml in a serum-separator (SST)-tube. In total, 460.8 ml of blood was withdrawn from each participant.
4
Middle Cerebral Artery Occlusion in Rodents
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The animals were randomly assigned into two groups for surgery; (i) sham MCAo (n=5) and (ii) MCAo (n=30). Right middle cerebral artery occlusion (MCAo) was performed under anaesthesia with Hypnorm/Dormicum (Fluanizon/Fentanyl/Midazolam) using the intraluminal filament technique [25, 26] . The animals remained anaesthetized for 60 min when the monofilament was withdrawn to allow for reperfusion. Xylocaine spray 10mg/ml (Astra Zeneca) was topically applied to the incision before suturing and returning the animals to their cages, where they were kept under observation and allowed to wake up.
Post-operative food intake was supported by administration of pellets soaked in Nutridrink (Nutricia).
Post-operative food intake was supported by administration of pellets soaked in Nutridrink (Nutricia).
5
Postprandial Liver Hemodynamics Study
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On the study day (Figure 1 ), a peripheral catheter was placed in the cubital vein, and baseline blood samples (Timepoint 0) were collected. After randomization, participants allocated to “postprandial” consumed a liquid meal consisting of 200 mL Nutridrink, Nutricia (300 kcal, 36.8 g carbohydrates, 11.8 g proteins, 11.6 g fat). Participants allocated to “fasting” remained fasting. Peripheral blood samples were collected after 15, 45, 60, 90, and 120 min. Transjugular liver vein catheterization was performed in all study participants to obtain liver biopsies and to measure hepatic venous pressure gradients (HVPG). At 60 min, a blood sample from the right liver vein was collected.
6
Postprandial PET Imaging of Bariatric State
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Participants ingested a 250 kcal liquid meal solution (Nutridrink, Nutricia Advanced Medical Nutrition, Amsterdam, Netherlands), consisting of 40 g of carbohydrates, 6 g of fat and 9 g protein, in 10 min. PET scans were repeated twice post ingestion: at 20 and 50 min for 15O-water, and at 40 and 70 min for 15O-carbon monoxide, respectively. These time points were chosen to cover accelerated gastric emptying rate, intestinal transit and peak GIP and GLP-1 concentrations in the post-bariatric state.
7
Functional MRI of Food Cue Reactivity
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The study consisted of one MRI scan session conducted in the morning. Subjects were scanned after an overnight fast (≥ 10h) after consumption of an ad libitum test meal (a commercially available drink called Nutridrink from Nutricia, see S2 Table for more details). They provided hunger and fullness ratings before and after test meal consumption. These served to ensure that their hunger decreased after test meal consumption. The amount of protein shake consumed ranged from 117–631 ml (Mean, SD = 449.7 ± 170.9 ml). Hunger ratings decreased and fullness ratings increased significantly after protein shake consumption (9-point Likert scale measurements: pre-meal hunger (Mean, SD) = 6.1±1.8; after meal hunger (Mean, SD) = 2.1 ± 1.1; and pre-meal fullness (Mean, SD) = 2.5±1.2; after meal fullness (Mean, SD) = 7.2 ± 1.5).
Before the scan, participants conducted a computerized food picture rating task (based on [31 (link)]). Subsequently, the participants underwent a 30-min MRI scan session. The first functional run consisted of a food and non-food viewing task, the second consisted of a forced choice task. In this paper we report the results of the forced choice task (seeFig 1 ).
Before the scan, participants conducted a computerized food picture rating task (based on [31 (link)]). Subsequently, the participants underwent a 30-min MRI scan session. The first functional run consisted of a food and non-food viewing task, the second consisted of a forced choice task. In this paper we report the results of the forced choice task (see
8
Circadian Rhythm and Metabolic Assessment
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Full details on the 24-hour blood sampling procedure have been described previously [36 (link)]. In short, a catheter was placed in a vein of the forearm of the nondominant hand and blood was collected every 10 minutes starting around 09:00 am . The participants received standardized feeding consisting of 600 kcal Nutridrink (Nutricia Advanced Medical Nutrition, Zoetermeer, The Netherlands) at 3 fixed times during the day (between 9:00 and 10:00 am , 12:00 pm and 1:00 pm , and 6:00 pm and 7:00 pm ). Lights were switched off for approximately 9 hours (circa between 11:00 pm and 8:00 am ) to allow the participants to sleep. Height and weight were measured in the research center. Body mass index (BMI) was calculated as weight (in kilograms) divided by the square of height (in meters). Body composition was determined by Bioelectrical Impedance Analysis at a fixed frequency of 50 kHz (Bodystat 1500 Ltd, Isle of Man, UK). Waist circumference was measured with a measuring tape midway between the uppermost border of the iliac crest and the lower border of the costal margin. The Pittsburgh Sleep Quality Index questionnaire was used to collect data on habitual bedtime and getting up time during the past month [37 (link)].
9
Circadian Rhythms in Blood Markers
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Participants were admitted to the research center at 0800 h, where a catheter was placed in a vein of the forearm of the non-dominant hand. After approximately an hour rest, blood sampling started at 0900 h. During 24 h, every 10 minutes 1.2 mL of blood was collected in a K3-EDTA tube and 2 mL in a serum separator (SST) tube. In total 461 mL of blood was withdrawn from each participant. All participants received standardized feeding at three fixed times during the day (between 0900–1000 h, 1200–1300 h, and 1800–1900 h), each consisting of 600 kcal Nutridrink (Nutricia Advanced Medical Nutrition, Zoetermeer, The Netherlands). Light exposure was standardized and lights were switched off between 2300–0800 h. No naps were allowed and participants ambulated only to the bathroom. All participants were sampled in the same room.
10
Dietary fat-induced obesity model
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The animals (n = 12–16 per group) were placed after weaning on a high fat diet (60% kcal fat, D12492, Research Diets) for 15 weeks. After becoming diet-induced obese, the animals were treated once daily, 1 hour prior to lights off, by intra-gastric gavage with either denatonium benzoate (60 μmol/kg, Sigma-Aldrich) or quinine-HCl (160 μmol/kg, Fagron) or water as control, during 4 weeks. At the day of sacrifice, the animals were fasted for 6 hours, before receiving a test meal (Nutridrink®, Nutricia) by gavage. The animals were anaesthetized with a mixture of xylazine and ketamine, 10 min after receiving the test meal, and sacrificed by cardiac exsanguination and decapitation. Blood samples were supplemented with 1 mM EDTA, 4 mM AEBSF (Sigma-Aldrich) and dipeptidyl peptidase 4 inhibitor (10 μl/ml, Millipore) and were centrifuged. Plasma samples and relevant tissues were collected and stored appropriately for further analysis. The hypothalamus was dissected as a whole using a mouse brain matrix (Zivic instruments).
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