With the animals under general anesthesia, endoscopy was performed. Animals were allowed a liquid diet 48 hours before the procedure and water only ad libitum 24 hours before the procedure. Antibiotics were administered for 5 days after the procedure (ceftiofur 5 mg/kg IM daily and metronidazole 1 g bid PO). Analgesia

(buprenorphine hydrochloride 0.03 mg/kg IM) was given immediately after the procedure. Animals were placed on a liquid diet for 1 day after the procedure, fed softened food on the second day, and by the third day, the animals resumed regular feed if tolerated. Each animal received oral proton pump inhibitors (Nexium [esomeprazole magnesium] 40 mg bid PO) for 7 days after the procedure. After PLX4032 order a 2-week

survival period, repeat endoscopy was performed. Animals were chemically euthanized (pentobarbital 100 mg/kg IV) immediately after endoscopy, and this was followed by necropsy. The intent was to create a submucosal tunnel within which a full-thickness biopsy specimen that included the muscularis propria would be obtained. The resection site was offset from the mucosal entry point to the submucosal tunnel http://www.selleckchem.com/products/AZD2281(Olaparib).html by approximately 4 to 5 cm. The overlying mucosal flap created by tunneling through the submucosa was used as a sealant flap protecting the peritoneum from contamination by the gastric contents. A large submucosal fluid cushion (SFC) was initially formed by using saline solution (∼40 mL) injected via a standard needle injection catheter (23-gauge Injector Force; Olympus America, Center Valley, Pa). A small incision (<5 mm) was made on the proximal aspect of the SFC by using a needle-knife, which served as the mucosal entry point. A tunneling balloon 18 mm in diameter (Apollo Endosurgery Inc, Austin, Tex) was inserted in the SFC, and as the balloon was inflated (Fig. 1), the unique Mirabegron progressive unfurling of this dilation balloon created a submucosal tunnel revealing muscularis propria. The length of the submucosal tunnel varied and

depended on the length of the balloon used (5-8 cm) and degree of balloon inflation. After the submucosal tunnel was created, a double-channel endoscope (2T 160; Olympus America) with an EMR-type cap attached was advanced through the submucosal tunnel. The EMR clear cap maintained tunnel patency and allowed improved visualization. An endoscopic Doppler probe (VTI Vascular Technology, Nashua, NH) was advanced through the endoscope working channel and placed within this submucosal space to identify any underlying blood vessels. Then a spiral tissue helix (Apollo Endosurgery Inc) or rat-tooth grasping forceps (Olympus America) was used to tent the muscularis propria toward the endoscope and into the cap. By using electrocautery, the muscularis propria was resected by using a spiral snare (Olympus America) or hexagonal snare (Traxtion US Endoscopy, Mentor, Ohio). Tissue was retrieved and submitted for analysis.