Flow cytometric measurement of cellular FLICE-inhibitory protein (c-FLIP) in ovarian carcinoma effusions.

OBJECTIVE: The objective of this study was to establish a flow cytometry assay for measuring c-FLIP in serous effusions. In addition, we studied the clinical relevance in ovarian carcinoma effusions of this inhibitor protein in the death receptor signalling pathway of apoptosis. METHODS: Two c-FLIP antibodies were tested using Western blotting and the best performing one was used for titration of c-FLIP expression in a panel of five cell lines, consisting of ovarian carcinoma, breast carcinoma and malignant mesothelioma. The concentration that provided the best signal-to-noise ratio was used for comparison of the performance of three fixation and permeabilization protocols. The best performing protocol was chosen for analysis of 69 ovarian carcinoma effusions. c-FLIP expression was analysed for association with clinicopathological parameters and survival. RESULTS: Rabbit polyclonal c-FLIP by Abcam and the IntraStain kit by Dako performed best. c-FLIP expression was detected in tumour cells in all 69 effusions (expression range 21-100%, median = 80%). No association was found between c-FLIP expression and clinicopathological parameters, including chemoresponse and survival. However, an inverse correlation was found between c-FLIP levels and expression of the previously studied apoptosis marker cleaved caspase-3 (P = 0.029). CONCLUSIONS: An assay for measuring c-FLIP in cytology specimens is presented. c-FLIP is frequently expressed in ovarian carcinoma effusions, but its expression appears to be unrelated to disease aggressiveness.

Intravenous etomidate for procedural sedation in emergency department patients.

OBJECTIVE: To determine the safety and effectiveness of intravenous (IV) etomidate for the sedation of patients undergoing painful procedures in the emergency department (ED). METHODS: A two-part feasibility study for ED patients receiving IV etomidate for painful ED procedures was undertaken. In the initial phase, a retrospective series of patients receiving etomidate for ED procedural sedation was considered. This phase served as the basis for a descriptive, prospective feasibility study of consecutive ED patients. During the second phase, patients were evaluated for complications related to IV etomidate sedation or the procedure performed. Immediately following the procedure, the physician was asked to complete a data collection sheet documenting the patient's etomidate dose, the number of doses required to complete the procedure, the analgesic used, the complications of the procedure, and the patient's procedural recall. RESULTS: Intravenous etomidate was administered to nine patients during the initial study phase and 51 during the prospective, descriptive phase. Indications for sedation included dislocation reduction (25), cardioversion (7), fracture reduction (20), abscess incision and drainage (4), foreign body removal (3), and chest thoracostomy (1). Physicians used 0.1-mg/kg IV bolus etomidate. A mean of 1.6 doses of etomidate was used to complete procedures (range 1-3 doses). Of the 60 patients in both study groups, 59 (98%) achieved adequate sedation by physician's assessment. Procedural success was documented for 56 patients (93%). There were 12 complications reported: oxygen desaturation below 90% (5), myoclonus (4), vomiting (1), pain with injection (1), and a brief bradycardic episode (1). No patient required assistance with ventilation or endotracheal intubation. CONCLUSIONS: Intravenous etomidate can be administered safely and effectively to provide appropriate conscious sedation for short, painful ED procedures.