Expression of the antibacterial peptide, cecropin, in cultured mammalian cells

We have stably transfected a Chinese hamster lung cell line V79 with a recombinant gene construct where the Drosophila cecropin A2 cDNA is under the control of Rous sarcoma virus, long terminal repeat (RSV LTR). We have not only been able to demonstrate expression at the RNA level by Northern analysis but also have detected an unprocessed peptide using an antiserum raised against Hyalophora cecropin.     

Antibiotic susceptibility testing ofCandida albicans by flow cytometry

A flow cytofluorometric susceptibility test (FCST) for in vitro antifungal drug testing ofCandida albicans was developed. Membrane damage was indicated by increased cellular fluorescence owing to propidium iodide or rose bengal uptake. Ketoconazole caused an exponential dose-response effect, best defined by E_max, at therapeutically achievable concentrations (0.02–0.2 µg/ml). This effect was not comparable to the conventional minimum inhibitory concentration (MIC) effect elicited by higher antibiotic concentrations. Amphotericin B, on the other hand, did not elicit E_max, but caused a >1 log dose-response effect which did correspond to the MIC. 5-Fluorocytosine susceptibility was also measurable. Other cytometric data indicating abnormal growth and growth inhibition, as well as conventional growth inhibition testing, confirmed that the 10-h FCST measured useful parameters of in vitro susceptibility.