Chemiluminescent assays have been used to quantify phagocytic activity since 1972. In recent years these assays have been adapted to the 96-well microplate format as new luminometers have been developed. In this report we describe the optimization of a lucigenin enhanced phagocyte chemiluminescent assay using a Titertek Luminoskan. Factors such as cell concentration, serum concentration in the opsonization of the zymosan used and lucigenin concentration were all optimized in our assay. In addition we have found that some of the unique features of the Luminoskan, continuous microplate agitation during the assay and microplate temperature control up to 43 degrees C, also significantly enhanced the chemiluminescent response.
Personal protection against infectious or toxic aerosols is a growing area of concern among many different occupational groups. Selection of appropriate respiratory protection should be based upon the level of risk present and the degree of protection afforded by the different types of protective masks available. In this study we have determined the relative protection factors for various commercial and military respiratory protective devices when challenged with a polydispersed submicron to micron sized particulate aerosol.
The effects of liposome-encapsulated ciprofloxacin on phagocytosis, nitric oxide production and intracellular killing of Staphylococcus aureus in murine macrophages were evaluated in this study. Mice were pretreated with three daily doses of liposome-encapsulated ciprofloxacin (45 mg/kg body weight/dose, intraperitoneal injection). At day 3 post drug administration, peritoneal macrophages were harvested by peritoneal lavage, and the phagocytic activity of the macrophages was determined by a chemiluminescence assay using opsonized zymosan particles. The phagocytic activity was found to be 7-fold higher in the liposome-encapsulated ciprofloxacin-treated group when compared to the untreated control group. For S. aureus-infected macrophages incubated with liposomes containing subinhibitory concentrations of ciprofloxacin (0.05 to 0.25 microg/mL), there were significant increases (up to 40 microM) in the levels of nitrite (NO2-, an end product of nitric oxide synthesis), and concommitant decreases (2-3 log) in the intracellular concentrations of S. aureus. Peak nitrite levels (20-40 microM) were produced when concentrations of liposome-encapsulated ciprofloxacin used were 0.1 to 0.25 microg/mL. These results suggest that liposome-encapsulated ciprofloxacin may have profound effects on the immunological functions of macrophages.
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