Six weeks after surgery, the rate of gastric emptying was assessed as previously described (Chambers et al., 2014). Briefly, after an overnight fast, 50% dextrose mixed with acetaminophen (100mg/kg, Sigma-Aldrich) was delivered orally. Blood was collected from the tail vein at 15 min after gavage. Plasma acetaminophen levels were measured using spectrophotometry (Sekisui Diagnostics).
Acetaminophen
Manufactured by Sekisui
Acetaminophen is a laboratory reagent used in the analysis and testing of various chemical compounds. It is a common analgesic and antipyretic compound. The core function of acetaminophen is to serve as a reference standard or calibration material in analytical procedures.
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Lab products found in correlation
Acetaminophen, by Merck Group (8 mentions)
Spectrophotometry assay, by Sekisui (4 mentions)
Biosen glucose analyzer, by EKF Diagnostics (2 mentions)
Ensure plus, by Merck Group (2 mentions)
Accu chek, by Roche (2 mentions)
Accu chek aviva plus, by Roche (2 mentions)
Ferritin, by Abcam (2 mentions)
Iron assay kit, by Abcam (2 mentions)
Fgf21, by R&D Systems (2 mentions)
Erythropoietin, by Abcam (2 mentions)
Igf 1, by R&D Systems (2 mentions)
Total glp 1, by Mesoscale (2 mentions)
Hemoglobin, by Abcam (2 mentions)
Fgf23, by Abcam (2 mentions)
Glycogen assay kit, by Merck Group (2 mentions)
Activin a, by R&D Systems (2 mentions)
Insulin, by Crystal Chem (2 mentions)
6 protocols using acetaminophen
1
Gastric Emptying Assessment Post-Surgery
2
Glucose Tolerance and Gastric Emptying
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Body weight was monitored biweekly. IPGTT was performed by intraperitoneal injection of 50% dextrose (2 g/kg) in 4-hour fasted mice. MMTT was performed via an oral gavage of liquid meal (volume 200 μL Ensure Plus spiked with a 40 mg dextrose and 4 mg acetaminophen, MilliporeSigma) in 4-hour fasted mice. Blood was obtained from the tail vein, and blood glucose was measured with Biosen Glucose Analyzer (EKF Diagnostics). Blood was collected at baseline and 15 minutes after gavage in EDTA-coated microtubes. Plasma acetaminophen levels were used to assess the rate of gastric emptying and were measured using spectrophotometry assay (Sekisui Diagnostics).
3
Glucose Tolerance and Gastric Emptying Evaluation
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Body weight was monitored monthly for 9 weeks prior and 12 weeks after Sham/VSG surgery. Intraperitoneal glucose tolerance test (IPGTT) was performed by intraperitoneal (IP) injection of 50% dextrose (2 g/kg) in 4-h fasted mice. Mixed-meal tolerance test (MMTT) was performed via an oral gavage of liquid meal (volume 200 µl Ensure Plus spiked with a 40-mg dextrose and 4-mg acetaminophen, Sigma-Aldrich) in 4-hour fasted mice. Blood was obtained from the tail vein and blood glucose was measured with Accu-Chek blood glucose meter (Accu-Chek Aviva Plus, Roche Diagnostics). Blood was collected from the tail vein at baseline and 15 min after gavage in EDTA-coated microtubes. Plasma acetaminophen levels were used to assess the rate of gastric emptying and were measured using spectrophotometry assay (Sekisui Diagnostics).
4
Intestinal Lipid and GLP-1 Analysis
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Metabolic studies, blood collection, fast protein liquid chromatography, and euthanization were as previously described. 29 (link) Briefly, the small intestine was divided into duodenum, jejunum, ileum (1:3:2 ratio), and flushed with 0.5 mmol/L sodium taurocholate (37 °C) followed by ice-cold saline. Whole intestinal tissue was used for analysis unless otherwise indicated. Plasma lipids, insulin, 26 (link) tissue lipids, 29 (link) and gene expression 27 (link) were measured as previously described. For GLP-1 measurement (total-K1503PD- Triglyceride values in the chylomicron and VLDL fraction were corrected for plasma volume spun and respective fraction volume collected. In WT mice, plasma triglyceride was measured using the Infinity Triglycerides assay. For gastric emptying measurements, plasma acetaminophen was measured (Sekisui Diagnostics, Charlottetown, PE).
5
Comprehensive Metabolic Profiling in VSG
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Insulin (Crystal Chem), total GLP-1 (MesoScale Discovery), and acetaminophen (Sekisui Diagnostics) were measured during experiments shown in Fig. 3 . Insufficient amount of blood was obtained from one Control VSG for Insulin, total GLP-1 and acetaminophen levels at 15 min. Postprandial plasma obtained at termination of studies (see above for details) was used to measure IGF-1 (R&D Systems), Activin-A (R&D Systems), Myostatin (GDF8) (R&D Systems), FGF21 (R&D Systems), L-amino acids (Abcam), Erythropoietin (Abcam), FGF23 (Abcam), Ferritin (Abcam), Hemoglobin (Abcam), Phosphate (Abcam), 25(OH) Vitamin D (Abcam). Iron levels in cecal contents were measured with Iron Assay Kit (Abcam). Glycogen content was measured in liver and muscle (tibialis anterior) samples with Glycogen Assay kit (Sigma-Aldrich). All sampled blood was collected via tail vein in EDTA-coated tubes. All assays were performed according to the manufacturer’s instructions.
6
Acetaminophen Absorption Assay for Gastric Emptying
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The acetaminophen absorption test was used to assess the rate of gastric emptying. After an overnight fast, mice were gavaged with a solution that contained 1% (w/v) acetaminophen (Sigma–Aldrich) in water. The dose of acetaminophen administered was 100 mg/kg. Tail vein blood (50 μL) was collected into heparin-coated tubes at 0, 15, 30, and 60 min after acetaminophen administration. Plasma was separated by centrifugation at 4 °C and stored at −80 °C until measurement of acetaminophen levels using an enzymatic-spectrophotometric assay (Sekisui Diagnostics, Charlottetown, PE).
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