Ketamine xylazine

Tracheas were transplanted as previously described from our group [19 (link), 24 (link)]. Donor tracheal segments were heterotopically transplanted into C57BL/6 recipients. For allografts, BALB/c tracheas were transplanted into C57BL/6 recipients. Both recipient and donor were C57BL/6 mice for isograft controls. Briefly, Tracheas were resected from freshly euthanized donor mice. The tracheas were immediately placed in ice-cold PBS with penicillin G sodium (100 U/ml) and streptomycin sulfate (100 μg/ml) (Life Technologies). C57BL/6 recipient mice were anesthetized with ketamine/xylazine (100 and 2 mg/kg intraperitoneally; Phoenix Pharmaceuticals, St. Joseph, MO). A 0.5 cm × 0.5 cm cross-shaped incision was made through the skin on the back of the recipient mouse. Four subcutaneous pockets were formed by blunt dissection. One tracheal graft was placed heterotopically into each subcutaneous pocket and incisions closed with suture. No immunosuppressive agents were given to any graft recipient.

Twelve-week-old male and female DSS were used in the present study (Envigo, Prattville, AL). All experiments were conducted in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals and were approved and monitored by the Augusta University Institutional Animal Care and Use Committee. Rats were housed in temperature- and humidity-controlled light-cycled quarters and maintained on either a normal-fat diet (NFD; F4031, Bio-Serv, Flemington, NJ) or HFD (F3282, Bio-Serv) from 12 to 16 wk of age (4-wk treatment period). The control NFD consisted of 3.88 kcal/g of gross energy with calories from the following sources: 20.5% protein, 61.6% carbohydrates, and 7.2% fat. The HFD consisted of 5.45 kcal/g of gross energy with calories from the following sources: 20.5% protein, 35.7% carbohydrates, and 36.0% fat. Both diets contain 0.8% NaCl. At the end of all experiments, rats were anesthetized with ketamine-xylazine (50 mg/kg and 6 mg/kg intraperitoneally, respectively, Phoenix Pharmaceuticals, St. Joseph, MO), a thoracotomy was performed, a terminal blood sample was obtained by aortic puncture, and tissues were harvested for flow cytometric analysis of T cells and cytokines.

Male and female SHR and WKY (Harlan Laboratories, Indianapolis, IN) were studied. All experiments were conducted in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals, European Union Directive 2010/63/EU for animal experiments and approved and monitored by the Georgia Regents University IACUC. Rats were housed in temperature and humidity‐controlled, light‐cycled quarters and maintained on a standard rodent diet (Teklad 8604, Madison, WI). At 12–13 weeks of age, rats were anesthetized with ketamine/xylazine (50 mg/kg and 6 mg/kg ip, respectively; Phoenix Pharmaceuticals, St. Joseph, MO) and thoracic aortas and third‐order mesenteric arteries were isolated.