The effect of sanguinarine on the RPMI-7951 and A375 melanoma cell lines

Background: Considering the resistance of melanoma to standard treatment protocols, the possibility of metastasis and the high mortality risk, the selection of new alternatives seems to be necessary. Compounds of natural origin are a promising option in anti-cancer therapy. One of them — sanguinarine has a wide spectrum of pro-health properties. Thus, the study aimed to assess the effect of the alkaloid on selected melanoma cellular models. Material and methods: Two types of melanoma cell lines were used in the study — A375 and RPMI-7951. The cells were treated with sanguinarine at concentrations ranging from 0.1 to 2 μM for 24 and 48 h. The influence of the alkaloid on such processes as cell death, cell cycle, organization of the main cytoskeletal proteins and migration potential was assessed. In addition, the sensitivity of selected cell lines to sanguinarine was evaluated based on the MTT assay. Results: The results showed that sanguinarine caused a dose-dependent decrease in cell survival compared to the untreated control. Further studies confirmed that it resulted from the pro-apoptotic and anti-proliferative action of the alkaloid. There were also significant changes in the organization of cytoskeletal proteins and the number of cells visible after fluorescent labelling. Moreover, in A375 cells, characteristics of entosis and mitotic catastrophe were noted. Sanguinarine-induced impaired cell migration was also confirmed. Conclusion: According to the authors’ knowledge, they are the first to present the influence of sanguinarine on the basic life processes of the RPMI-7951 cell line and supplement the knowledge regarding the A375 melanoma cells. The present results confirm the anticancer properties of the alkaloid (cytotoxicity, anti-migratory and pro-apoptotic effect)

Conventional and Real-Time PCR Targeting <i>bla</i>_OXA Genes as Reliable Methods for a Rapid Detection of Carbapenem-Resistant <i>Acinetobacter baumannii</i> Clinical Strains

Multidrug-resistant Acinetobacter baumannii, particularly those producing carbapenemases, are spread worldwide. A reliable detection of carbapenemases is essential to choose the appropriate antimicrobial therapy and, consequently, prevent the dissemination of carbapenem-resistant strains. The aim of this study is to examine the molecular basis of the carbapenem resistance mechanism and estimation of conventional PCR and real-time PCR usefulness for the detection of oxacillinases when compared to phenotypic carbapenemases detection. The following methods were evaluated: the CarbAcineto NP test, Carbapenem Inactivation Method, CPO panels of semiautomated antimicrobial susceptibility testing method on the BD Phoenix™ M50 system, conventional Polymerase Chain Reaction and real-time PCR. The eazyplex® SuperBug complete A assay was used as the reference method. Among the tested strains, 39 (67.2%) carried the blaOXA-40 gene, while the blaOXA-23 gene was noted amongst 19 (32.8%) isolates. The diagnostic sensitivities of the studied assays were as follows: CarbAcineto NP—65.5%; CIM—100%; CPO—100%; conventional PCR—100%; real-time PCR—100%