A separate surgical study cohort who had RYGB surgery at Imperial Weight Centre between 2007 and 2012 was also recruited for measurement of the reference gut hormone levels we would be targeting in our infusion study. The RYGB volunteers arrived at the unit at 9 am in the fasted state. A standardized mixed meal test (Ensure Plus; Abbott, Maidenhead, United Kingdom; 13.8 g of protein, 10.8 g of fat, 44.4 g of carbohydrates, 330 kcal, 220 mL) was given, and volunteers were directed to drink the Ensure Plus over 10 minutes. Blood was taken from an indwelling cannula placed in the antecubital fossa for gut hormone (GLP-1, OXM, and PYY), glucose, and insulin measurement at baseline and 30, 60, and 120 minutes post Ensure Plus ingestion.
Ensure plus
Manufactured by Abbott
Sourced in United States, United Kingdom, Belgium
Ensure Plus is a nutritional supplement drink manufactured by Abbott. It is designed to provide balanced nutrition, including protein, carbohydrates, and essential vitamins and minerals. The product is intended to be a convenient source of nutrition for individuals who may have difficulty meeting their dietary requirements through regular food intake.
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Lab products found in correlation
Vacutainer edta tube edta aprotinin tube, by BD (2 mentions)
Intralipid, by Merck Group (2 mentions)
Sucrose solution, by Avantor (2 mentions)
Neuroscience studio software, by Doric (1 mentions)
Prosure, by Abbott (1 mentions)
Acetaminophen, by Merck Group (1 mentions)
Ensure plus, by Merck Group (1 mentions)
Lipid extraction kit, by Abcam (1 mentions)
Proteome profiler mouse xl cytokine array, by R&D Systems (1 mentions)
Dcfda h2dcfda assay kit, by Abcam (1 mentions)
Mouse insulin elisa kit, by Crystal Chem (1 mentions)
Oxiselect tbars assay kit, by Cell Biolabs (1 mentions)
Oxyntomodulin, by Phoenix Pharmaceuticals (1 mentions)
Aprotinin, by Merck Group (1 mentions)
Glp 2, by Phoenix Pharmaceuticals (1 mentions)
Ek 059 02, by Phoenix Pharmaceuticals (1 mentions)
Ek 028 11, by Phoenix Pharmaceuticals (1 mentions)
3h triolein, by PerkinElmer (1 mentions)
Microplate reader, by Tecan (1 mentions)
41 protocols using ensure plus
1
Postprandial Gut Hormone Dynamics in RYGB
2
Bariatric Nutrition Protocol for DJBL
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Subjects were studied on three occasions: (1) within 1 month prior to implantation, (2) 1 week after implantation, and 3) 24 weeks after implantation, just prior to removal. After implantation of the DJBL, subjects were provided a standard of care bariatric nutritional counseling program, which suggested a regular diet with a maximum of 1200 kcal for women and 1500 kcal for men. For safety reasons, liquid nutrition was advised for the first week after DJBL placement. At every visit, body weight was determined and subjects were asked if their satiety and caloric intake had increased, decreased, or was unchanged when compared to baseline. Daily caloric intake was evaluated by a dietitian and if necessary, dietary adaptations were made. Additionally, a standardized meal tolerance test was performed: venous blood samples were drawn after an overnight fast; subsequently a standard liquid meal was consumed (Ensure Plus, Abbott Laboratories, IL; 333 mL, 500 kcal, 20.8 g protein, 67.3 g carbohydrates, and 16.4 g fat), followed by collection of blood samples after 10, 30, 60, 90, and 120 min (BD Vacutainer EDTA tube/EDTA aprotinin tube, BD diagnostics, Erembodegem-Aalst, Belgium). Samples were immediately cooled, centrifuged, and stored at -80 °C until analysis.
3
Defined Mixed Meal Protocol
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Having fasted overnight, each participant consumes a defined mixed meal (Ensure Plus®; Abbott Nutrition, Lake Forest, IL, USA) containing 57% of total calories from carbohydrates, 28% from fat, and 15% from protein. The amount of the mixed meal provided is calculated by a staff dietician to provide 30% of the participant’s daily energy requirement based on total body weight (Harris–Benedict equation) and fat free mass (kg) (Katch–McArdle formula) [22 (link)].
4
Optogenetic Stimulation of POMC→MeA Pathway
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We crossbred the POMC-Cre strain with the floxed-stop ChR2-tdTomato strain to generate POMC-Cre; ChR2-tdTomato mice. To stimulate the ARCPOMC→MeA projection, a mono fiber-optic cannula of 230 μm diameter (MFC_200/230–0.48_5 mm, Doric Lenses) was implanted into the MeA (AP, −1.58 mm; ML, −2.0 mm; DV, −5.0 mm) and coupled to a 473 nm diode-pumped solid-state (DPSS) laser (Laserglow Technologies). Animals were allowed at least a week to recover from surgery. ChR2-expressing POMC fibers in the MeA were illuminated at 20 Hz (25 ms pulse duration, 20 Hz pulses per 1 s, 3 s interval between events). Stimulation pulses were generated with Doric Neuroscience Studio software (Doric Lenses). Light illumination started 30 min before food presentation. To measure liquid food intake (Ensure Plus, Abbott), animals were separated individually in single cages for 5 days and fasted for 6 h (from 12 PM to 6 PM). The MC4R antagonist SHU-9119 (1 mg/kg) was intraperitoneally injected 30 min before food presentation.
5
Ischemic Enteritis and n-3 PUFA Therapy
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Ischemic enteritis was induced by controlled narrowing of the superior mesenteric artery, and following the rats were divided randomly into two groups: N3PUFA (n = 20) -Following surgery rats were solely fed with a high-molecular polymer artificial total enteral nutrition enriched with n-3 PUFA in a ratio n-6/n-3 = 0.3/1.0, containing eicosapentaenoic acid and docosahexaenoic acid for 15 days (ProSure ® , Abbott 240 ml). The detailed ingredients of the diet are shown in table 1. During the autopsy, blood and tissue samples were taken. Control (n = 20) -Control Group: Following surgery rats were solely fed with a highmolecular polymer artificial total enteral nutrition (Ensure Plus ® , Abbott 200 ml) containing alpha linolenic acid for 15 days. The detailed ingredients of the diet are shown in table 1. During the autopsy, blood and tissue samples were taken.
Pilot studies showed that the daily consumption of the nutrition formula was 20 ml/100 g body weight/day. Hence, the estimated levels of total n3 fatty acids in the N3PUFA group administrated were 0.13 g/100 g body weight/day, so each animal received approximately 0.195-0.26 g of n3 fatty acids per day and in the CONTROL group was 0.055 g/100 g body weight/ day, so each animal received approximately 0.0825-0.11 g of n3 fatty acids/day.
Pilot studies showed that the daily consumption of the nutrition formula was 20 ml/100 g body weight/day. Hence, the estimated levels of total n3 fatty acids in the N3PUFA group administrated were 0.13 g/100 g body weight/day, so each animal received approximately 0.195-0.26 g of n3 fatty acids per day and in the CONTROL group was 0.055 g/100 g body weight/ day, so each animal received approximately 0.0825-0.11 g of n3 fatty acids/day.
6
Oral and Intraperitoneal Glucose Tolerance Tests
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For the oral GTT, all animals were fasted for 6 h prior to an oral gavage of 200 ul Ensure Plus (Abbott Laboratories). For the i.p. GTT, all animals were fasted for 6 h prior to an i.p. injection of 25% dextrose at a dose of 8 ul/g. All glucose was measured by gluocometer prior to gavage/injection (time 0), and then 15, 30, 60, and 120 min post gavage/injection. Blood was collected from the tip of the tail by cutting a small amount of the tail and gently massaging the blood out. Animals were excluded from analysis based on pre-established criteria, i.e. if blood glucose did not rise significantly over the 2-h period, indicating a failed glucose injection.
7
Oral and Intraperitoneal Glucose Tolerance Tests
8
Mixed Meal Tolerance Test for Insulin Sensitivity
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MMTT was performed to calculate indices of insulin sensitivity. During MMTT, a liquid meal (Ensure©Plus, Abbott Nutrition) was given at 6 kcal/kg (max 360 kcal). The liquid meal consisted of approximately 30% of energy from fat, 15% energy from protein, and 55% energy from carbohydrates. Blood samples were collected before and at 30, 60, 90, and 120 minutes after MMTT for plasma glucose and insulin measurements.
9
Acetaminophen-Enriched Ensure Plus Meal
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Ensure Plus (Abbott Labs, USA; Per 8 oz [237 mL] bottle consists of total fat [11 g], cholesterol [0.01 g], total carbohydrates [51 g containing 1 g dietary fiber and 22 g sugar), protein [13 g] and various vitamins plus minerals) was used for the MMTT as previously reported34 . Acetaminophen (Sigma-Aldrich, USA) was slowly added to Ensure Plus solution (4 mg/mL) and continuously stirred for 6 h. The prepared meal solution containing Acetaminophen was kept at room temperature (22 °C) and protected from light until being used within 24 h. The meal solution was continuously mixed with a magnetic stir-bar at least 30 min before dosing.
10
Assessing Endogenous Insulin Production
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The endogenous insulin production in the participants was assesed by measuring stimulated serum C-peptide levels at study start (defined as baseline) and 6 and 12 months after inclusion. Blood was drawn at fasting and 90 minutes after ingestion of a standardized liquid mixed-meal (Boost High Protein, Nestlé Health Science, or Ensure Plus, Abbott Nutrition). Plasma and serum were isolated accordingly and samples were stored at − 80˚C until analysed. C-peptide and HbA1c were measured using an Immulite® 2000 chemiluminescent immunometric assay and high-pressure liquid chromatography (Tosoh Bioscience, South San Francisco, CA, USA), respectively.
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