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      Glabridin triggers over-expression of apoptosis inducing factor (AIF) gene in Candida albicans

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          Abstract

          Background and Purpose:

          Candida albicans is a prevalent human fungal pathogen that can cause a wide spectrum of diseases, from superficial mucosal infections to systemic disorders, in patients with impaired immunity. Glabridin is a pyranoisoflavan originally extracted from root extract of Glycyrrhiza glabra. Glabridin can also mediate apoptosis in yeast cells by changing the mitochondrial membrane potential, activation of caspase-like proteases, and DNA cleavage. The aim of this study was to investigate the mechanism of action of glabridin in C. albicans.

          Materials and Methods:

          Candida albicans ATCC14053 was applied as the standard strain. Total RNA was extracted from the isolate under glabridin-treated and untreated conditions. To evaluate the alternations in the apoptosis inducing factor ( AIF) gene expression, real-time polymerase chain reaction (real-time -PCR) was performed, and the obtained data were analyzed using REST software.

          Results:

          Expression of the AIF gene was represented as the ratio of expression relative to the reference gene. According to the REST ® output, the expression of the AIF gene increased significantly ( P<0.05) under the glabridin-treated condition.

          Conclusion:

          Our results suggested that glabridin may induce apoptosis through the caspase-independent route and might be considered as an anti- Candida agent.

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          Most cited references19

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          Candida albicans mutations in the ergosterol biosynthetic pathway and resistance to several antifungal agents.

          The role of sterol mutations in the resistance of Candida albicans to antifungal agents has not been thoroughly investigated. Previous work reported that clinical C. albicans strains resistant to both azole antifungals and amphotericin B were defective in ERG3, a gene encoding sterol Delta(5,6)-desaturase. It is also believed that a deletion of the lanosterol 14alpha-demethylase gene, ERG11, is possible only under aerobic conditions when ERG3 is not functional. We tested these hypotheses by creating mutants by targeted deletion of the ERG3 and ERG11 genes and subjecting those mutants to antifungal susceptibility testing and sterol analysis. The homozygous erg3/erg3 mutant created, DSY1751, was resistant to azole derivatives, as expected. This mutant was, however, slightly more susceptible to amphotericin B than the parent wild type. It was possible to generate erg11/erg11 mutants in the DSY1751 background but also, surprisingly, in the background of a wild-type isolate with functional ERG3 alleles under aerobic conditions. This mutant (DSY1769) was obtained by exposure of an ERG11/erg11 heterozygous strain in a medium containing 10 micro g of amphotericin B per ml. Amphotericin B-resistant strains were obtained only from ERG11/erg11 heterozygotes at a frequency of approximately 5 x 10(-5) to 7 x 10(-5), which was consistent with mitotic recombination between the first disrupted erg11 allele and the other remaining functional ERG11 allele. DSY1769 was also resistant to azole derivatives. The main sterol fraction in DSY1769 contained lanosterol and eburicol. These studies showed that erg11/erg11 mutants of a C. albicans strain harboring a defective erg11 allele can be obtained in vitro in the presence of amphotericin B. Amphotericin B-resistant strains could therefore be selected by similar mechanisms during antifungal therapy.
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            Adaptive immune responses to Candida albicans infection

            Fungal infections are becoming increasingly prevalent in the human population and contribute to morbidity and mortality in healthy and immunocompromised individuals respectively. Candida albicans is the most commonly encountered fungal pathogen of humans, and is frequently found on the mucosal surfaces of the body. Host defense against C. albicans is dependent upon a finely tuned implementation of innate and adaptive immune responses, enabling the host to neutralise the invading fungus. Central to this protection are the adaptive Th1 and Th17 cellular responses, which are considered paramount to successful immune defense against C. albicans infections, and enable tissue homeostasis to be maintained in the presence of colonising fungi. This review will highlight the recent advances in our understanding of adaptive immunity to Candida albicans infections.
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              Farnesol-induced apoptosis in Candida albicans.

              Farnesol, a precursor in the isoprenoid/sterol pathway, was recently identified as a quorum-sensing molecule produced by the fungal pathogen Candida albicans. Farnesol is involved in the inhibition of germination and biofilm formation by C. albicans and can be cytotoxic at certain concentrations. In addition, we have shown that farnesol can trigger apoptosis in mammalian cells via the classical apoptotic pathways. In order to elucidate the mechanism behind farnesol cytotoxicity in C. albicans, the response to farnesol was investigated, using proteomic analysis. Global protein expression profiles demonstrated significant changes in protein expression resulting from farnesol exposure. Among the downregulated proteins were those involved in metabolism, glycolysis, protein synthesis, and mitochondrial electron transport and the respiratory chain, whereas proteins involved in folding, protection against environmental and oxidative stress, actin cytoskeleton reorganization, and apoptosis were upregulated. Cellular changes that accompany apoptosis (regulated cell death) were further analyzed using fluorescent microscopy and gene expression analysis. The results indicated reactive oxygen species accumulation, mitochondrial degradation, and positive terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL) in the farnesol-exposed cells concurrent with increased expression of antioxidant-encoding and drug response genes. More importantly, the results demonstrated farnesol-induced upregulation of the caspase gene MCA1 and the intracellular presence of activated caspases. In conclusion, this study demonstrated that farnesol promotes apoptosis in C. albicans through caspase activation, implying an important physiological role for farnesol in the fungal cell life cycle with important implications for adaptation and survival.
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                Author and article information

                Journal
                Curr Med Mycol
                Curr Med Mycol
                CMM
                Current Medical Mycology
                Iranian Society of Medical Mycology (Sari, Iran )
                2423-3439
                2423-3420
                September 2018
                : 4
                : 3
                : 19-22
                Affiliations
                [1 ] Invasive Fungi Research Center, Mazandaran University of Medical Sciences, Sari, Iran
                [2 ] Department of Medical Mycology, School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran
                [3 ] Faculty of Medicine, Sari Branch, Islamic Azad University, Sari, Iran
                Author notes
                [* ]Corresponding author: Mojtaba Nabili, Faculty of Medicine, Sari Branch, Islamic Azad University, Sari, Iran. Email: M.nabili2010@gmail.com
                Article
                10.18502/cmm.4.3.172
                6315205
                47fed5f0-ee48-4e10-861f-27253bc1079f
                © 2018, Published by Mazandaran University of Medical Sciences on behalf of Iranian Society of Medical Mycology and Invasive Fungi Research Center.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License, ( http://creativecommons.org/licenses/by/4.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 22 June 2018
                : 15 September 2018
                : 9 October 2018
                Categories
                Short Communication

                aif gene,apoptosis,candida albicans,glabridin
                aif gene, apoptosis, candida albicans, glabridin

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