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      Real-time Monitoring of Non-specific Toxicity Using a Saccharomyces cerevisiae Reporter System

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          Abstract

          Baker's yeast, Saccharomyces cerevisiae, is the simplest and most well-known representative of eukaryotic cells and thus a convenient model organism for evaluating toxic effects in human cells and tissues. Yeast cell sensors are easy to maintain with short generation times, which makes the analytical method of assessing antifungal toxicity cheap and less-time consuming. In this work, the toxicity of test compounds was assessed in bioassays based on bioluminescence inhibition and on traditional growth inhibition on agar plates. The model organism in both tests was a modified S. cerevisiae sensor strain that produces light when provided with D-luciferin in an insect luciferase reporter gene activity assay. The bioluminescence assay showed toxic effects for yeast cell sensor of 5,6-benzo-flavone, rapamycin, nystatin and cycloheximide at concentrations of nM to μM. In addition, arsenic compounds, cadmium chloride, copper sulfate and lead acetate were shown to be potent non-specific inhibitors of the reporter organism described here. The results from a yeast agar diffusion assay correlated with the bioluminescence assay results.

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          Most cited references 53

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          Microbial heavy-metal resistance.

          We are just beginning to understand the metabolism of heavy metals and to use their metabolic functions in biotechnology, although heavy metals comprise the major part of the elements in the periodic table. Because they can form complex compounds, some heavy metal ions are essential trace elements, but, essential or not, most heavy metals are toxic at higher concentrations. This review describes the workings of known metal-resistance systems in microorganisms. After an account of the basic principles of homoeostasis for all heavy-metal ions, the transport of the 17 most important (heavy metal) elements is compared.
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            The world of subinhibitory antibiotic concentrations.

            Although antibiotics have long been known to have multiple effects on bacterial cells at low concentrations, it is only with the advent of genome transcription analyses that these activities have been studied in detail at the level of cell metabolism. It has been shown that all antibiotics, regardless of their receptors and mode of action, exhibit the phenomenon of hormesis and provoke considerable transcription activation at low concentrations. These analyses should be of value in providing information on antibiotic side-effects, in bioactive natural product discovery and antibiotic mode-of-action studies.
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              Pathways of As(III) detoxification in Saccharomyces cerevisiae.

              Saccharomyces cerevisiae has two independent transport systems for the removal of arsenite from the cytosol. Acr3p is a plasma membrane transporter that confers resistance to arsenite, presumably by arsenite extrusion from the cells. Ycf1p, a member of the ABC transporter superfamily, catalyzes the ATP-driven uptake of As(III) into the vacuole, also producing resistance to arsenite. Vacuolar accumulation requires a reductant such as glutathione, suggesting that the substrate is the glutathione conjugate, As(GS)3. Disruption of either the ACR3 or YCF1 gene results in sensitivity to arsenite and disruption of both genes produces additive hypersensitivity. Thus, Acr3p and Ycf1p represent separate pathways for the detoxification of arsenite in yeast.
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                Author and article information

                Journal
                Sensors (Basel)
                Sensors (Basel)
                Sensors (Basel, Switzerland)
                Molecular Diversity Preservation International (MDPI)
                1424-8220
                October 2008
                16 October 2008
                : 8
                : 10
                : 6433-6447
                Affiliations
                [1 ] Department of Chemistry and Bioengineering, Tampere University of Technology, P.O. Box 541, FI- 33101 Tampere, Finland; E-Mails: anniina.kivisto@ 123456tut.fi (A.K.); matti.karp@ 123456tut.fi (M.K.)
                [2 ] Biosensing Competence Centre (BCC), Tampere University of Technology, P.O. Box 541, FI- 33101 Tampere, Finland; E-Mails: anniina.kivisto@ 123456tut.fi (A.K.); matti.karp@ 123456tut.fi (M.K.)
                [3 ] Department of Applied Chemistry and Microbiology Division of Microbiology, P.O. Box 56, FI-00014, University of Helsinki, Finland; E-Mails: marko.virta@ 123456helsinki.fi (M.V.)
                Author notes
                [* ] Author to whom correspondence should be addressed; E-Mails: anna-liisa.valimaa@ 123456tut.fi (A.L); Tel.: +358-3-3115-2968; Fax: +358-3-3115-2869
                Article
                sensors-08-06433
                10.3390/s8106433
                3707459
                © 2008 by the authors; licensee Molecular Diversity Preservation International, Basel, Switzerland.

                This article is an open-access article distributed under the terms and conditions of the Creative Commons Attribution license ( http://creativecommons.org/licenses/by/3.0/).

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