The dynamics of intracellular levels of superoxide and NO after cell treatment with NO synthase inhibitors were studied in human cells expressing various NOS isoforms: endotheliocytes and ECV-304 (eNOS) and carcinoma cells and HeLa-G63 (iNOS). Cytometric analysis of changes in the cell fluorescence intensity was carried out using superoxide and NO fluorescent indicators (dihydroethidene and DAF-2-DA, respectively). Intracellular levels of superoxide decreased in HeLa-G63 and ECV-304 cells after their incubation in medium with aminoguanidine, L-NAME, and D-NAME. Intracellular NO level decreased only in HeLa-G63 cells after incubation in medium with aminoguanidine and L-NAME, but not D-NAME. The level of NO returned to normal after 7-h culturing in inhibitor-free medium, while the level of superoxide increased and remained high throughout 3 generations. Incubation of cells with D-NAME did not increase the intracellular level of superoxide. Presumably, high prolonged generation of superoxide is a delayed result of inhibition of NO synthesis in HeLa-G63 cells.
The transfecting activity of linear lambda DNA is 100 times higher in calcium treated E. coli K12 (lambda i434) than in non-lysogenic strains: the levels of transfection are 1-2.10(7) and 1-2.10(5) infective centers per 1 mug of lambda DNA respectively. The high efficiency of lysogenic cells transfection is not due to the spontaneously liberated "helper" phage. Evidently, it is called forth by transfecting DNA-prophage recombination or/and by inhibition of nuclease activity in lysogenic cells. Both ring forms lambda DNA (supercoiled and open circles) show very low infectivity, if any, in calcinated cells.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.