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      Evaluation of the Cytotoxic Activity of the Usnea barbata (L.) F. H. Wigg Dry Extract

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          Abstract

          The secondary metabolites of lichens have proven to be promising sources of anticancer drugs; one of the most important of these is usnic acid, which is a phenolic compound with dibenzofuran structure that is responsible for the numerous biological actions of lichens of genus Usnea. As a result, in this study, we related to this phenolic secondary metabolite. The aim of the present study is the evaluation of the cytotoxic activity of Usnea barbata (L.) F. H. Wigg dry acetone extract (UBE). In advance, the usnic acid content was determined in various extracts of Usnea barbata (L.) F. H. Wigg: the liquid extracts were found in water, ethanol, acetone, and the dry acetone extract; the highest usnic acid quantity was found in the dry acetone extract. First, the cytotoxic action of UBE was assessed using Brine Shrimp Lethality (BSL) test; a significant lethal effect was obtained after 24 h of treatment at high used concentrations of UBE, and it was quantified by the high mortality rate of the Artemia salina (L.) larvae. Secondly, in vitro cytotoxicity of UBE was evaluated on human tongue squamous cells carcinoma, using CAL 27 (ATCC ® CRL-2095™) cell line. The most intense cytotoxic effect of UBE on CAL 27 cells was registered after 24 h; this response is directly proportional with the tested UBE concentrations. The obtained results have been reported regarding usnic acid content of UBE, and the data show that CAL 27 cells death was induced by apoptosis and high oxidative stress.

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          Measurement of superoxide dismutase, catalase and glutathione peroxidase in cultured cells and tissue.

          Cells contain a large number of antioxidants to prevent or repair the damage caused by reactive oxygen species, as well as to regulate redox-sensitive signaling pathways. General protocols are described to measure the antioxidant enzyme activity of superoxide dismutase (SOD), catalase and glutathione peroxidase. The SODs convert superoxide radical into hydrogen peroxide and molecular oxygen, whereas the catalase and peroxidases convert hydrogen peroxide into water. In this way, two toxic species, superoxide radical and hydrogen peroxide, are converted to the harmless product water. Western blots, activity gels and activity assays are various methods used to determine protein and activity in both cells and tissue depending on the amount of protein required for each assay. Other techniques including immunohistochemistry and immunogold can further evaluate the levels of the various antioxidant enzymes in tissues and cells. In general, these assays require 24-48 h to complete.
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            Revisiting the hallmarks of cancer.

            The hallmarks of cancer described by Hanahan and Weinberg have proved seminal in our understanding of cancer's common traits and in rational drug design. Not free of critique and with understanding of different aspects of tumorigenesis coming into clearer focus in the recent years, we attempt to draw a more organized and updated picture of the cancer hallmarks. We define seven hallmarks of cancer: selective growth and proliferative advantage, altered stress response favoring overall survival, vascularization, invasion and metastasis, metabolic rewiring, an abetting microenvironment, and immune modulation, while highlighting some considerations for the future of the field.
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              Brine shrimp: a convenient general bioassay for active plant constituents.

              A method, utilizing brine shrimp (Artemia salina Leach), is proposed as a simple bioassay for natural product research. The procedure determines LC (50) values in microg/ml of active compounds and extracts in the brine medium. Activities of a broad range of known active compounds are manifested as toxicity to the shrimp. Screening results with seed extracts of 41 species of Euphorbiaceae were compared with 9KB and 9PS cytotoxicities. The method is rapid, reliable, inexpensive, and convenient as an in-house general bioassay tool.
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                Author and article information

                Contributors
                Role: Academic Editor
                Role: Academic Editor
                Role: Academic Editor
                Role: Academic Editor
                Journal
                Molecules
                Molecules
                molecules
                Molecules
                MDPI
                1420-3049
                17 April 2020
                April 2020
                : 25
                : 8
                : 1865
                Affiliations
                [1 ]Departament of Microbiology and Immunology, Faculty of Dental Medicine, Ovidius University of Constanta, 7 Ilarie Voronca Street, 900684 Constanta, Romania; popovicivioleta@ 123456gmail.com (V.P.); badea_victoria@ 123456yahoo.com (V.B.)
                [2 ]Department of Pharmacognosy, Faculty of Pharmacy, Ovidius University of Constanta, 6 Capitan Al. Serbanescu Street, 900001 Constanta, Romania; adrianabucur@ 123456yahoo.com
                [3 ]Department of Cellular and Molecular Biology, Faculty of Pharmacy, Ovidius University of Constanta, 6 Capitan Al. Serbanescu Street, 900001 Constanta, Romania
                [4 ]Institute of Biological Research Iasi, branch of NIRDBS, 47 Lascar Catargi Street, 700107 Iasi, Romania; mihai.cosmin.teo@ 123456gmail.com (C.T.M.); gabrielacapraru@ 123456yahoo.com (G.V.)
                [5 ]Advanced Centre for Research and Development in Experimental Medicine (CEMEX), “Grigore T. Popa” University of Medicine and Pharmacy Iasi, 9-13 Mihail Kogalniceanu Street, 700259 Iasi, Romania
                [6 ]Department of Oral Rehabilitation, Faculty of Dental Medicine, Ovidius University of Constanta, 7 Ilarie Voronca Str., 900684 Constanta, Romania; drcaraiane@ 123456yahoo.com (A.C.); florinb.md@ 123456gmail.com (F.C.B.)
                Author notes
                [†]

                These authors contributed equally to this study.

                [‡]

                These authors contributed equally to this study.

                Author information
                https://orcid.org/0000-0001-9696-5728
                https://orcid.org/0000-0002-7750-9871
                https://orcid.org/0000-0001-8876-1901
                https://orcid.org/0000-0002-0945-5437
                https://orcid.org/0000-0002-0876-473X
                Article
                molecules-25-01865
                10.3390/molecules25081865
                7221659
                32316674
                0d8701a6-8765-4ee0-a80d-eb01c7647506
                © 2020 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 16 March 2020
                : 14 April 2020
                Categories
                Article

                usnea barbata (l.) f. h. wigg,dry extract,usnic acid,cytotoxic activity,brine shrimp,cal 27 cells

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