For single-dose safety evaluation studies, we administrated two doses of fluorocoxib A, 0.1 and , i.v. over 20 min to normal beagle research dogs. We used three dogs per treatment group, as this is a common number employed in dose-escalation studies for chemotherapy toxicity in this species.33 The medical history of all research dogs was carefully evaluated before enrollment in the study. Physical examinations for signs of potential drug toxicity were performed before, during, and daily for three days after fluorocoxib A administration. Physical assessment included capillary refill time, auscultation of heart and lungs, abdominal palpation, appetite, attitude, activity levels, and other health-related events. Hypersensitivity was evaluated by direct observation of each dog during treatment for clinical signs of an allergic reaction (facial swelling, flushing, urticaria, dyspnea, fever, scratching, and changes of heart rates). Signs of adverse events after fluorocoxib A administration were monitored on a daily basis including vomiting, diarrhea, depression, nausea, and increased salivation. Laboratory evaluations of blood and urine were carried out before and three days after compound administration to monitor signs of fluorocoxib A toxicity. Blood (3 ml) was collected into ethylenediaminetetraacetic acid (EDTA)- and heparin- treated tubes, and urine (up to 5 ml) was collected by cystocentesis or from the floor of the runs. Note that research dogs urinate ad libitum, and urine samples collected from the floor of the runs were used, when necessary, for analysis. Renal toxicity effects, such as casts, presence of urine glucose, and proteins, are not likely to be affected by contact with the floor for a short time period. The laboratory evaluation consisted of a complete blood count (CBC), serum chemistry profile, and complete urinalysis. The complete blood count evaluated total white blood cells (WBC) and hematocrit (HCT) in addition to absolute numbers of neutrophils, lymphocytes, monocytes, eosinophils, and platelets. The plasma chemistry profile evaluated blood urea nitrogen (BUN), creatinine, proteins (albumin, globulins, and total proteins), alanine aminotransferase (ALT), total bilirubin, glucose, and electrolytes. The complete urinalysis evaluated urine proteins, glucose, and ketones as well as a sediment examination and urine specific gravity. All laboratory tests were carried out at the Veterinary Medical Center of the University of Tennessee in Knoxville. Because our study evaluated the safety of a single-dose administration rather than a long-term multidose exposure to fluorocoxib A, euthanasia of the exposed dogs was not a part of the protocol.