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      Transcriptome sequencing revealed the inhibitory mechanism of ketoconazole on clinical Microsporum canis

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          Abstract

          Background

          Microsporum canis is a zoonotic disease that can cause dermatophytosis in animals and humans.

          Objectives

          In clinical practice, ketoconazole (KTZ) and other imidazole drugs are commonly used to treat M. canis infection, but its molecular mechanism is not completely understood. The antifungal mechanism of KTZ needs to be studied in detail.

          Methods

          In this study, one strain of fungi was isolated from a canine suffering with clinical dermatosis and confirmed as M. canis by morphological observation and sequencing analysis. The clinically isolated M. canis was treated with KTZ and transcriptome sequencing was performed to identify differentially expressed genes in M. canis exposed to KTZ compared with those unexposed thereto.

          Results

          At half-inhibitory concentration (½MIC), compared with the control group, 453 genes were significantly up-regulated and 326 genes were significantly down-regulated ( p < 0.05). Quantitative reverse transcription polymerase chain reaction analysis verified the transcriptome results of RNA sequencing. Gene ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes enrichment analysis revealed that the 3 pathways of RNA polymerase, steroid biosynthesis, and ribosome biogenesis in eukaryotes are closely related to the antifungal mechanism of KTZ.

          Conclusions

          The results indicated that KTZ may change cell membrane permeability, destroy the cell wall, and inhibit mitosis and transcriptional regulation through CYP51, SQL, ERG6, ATM, ABCB1, SC, KER33, RPA1, and RNP genes in the 3 pathways. This study provides a new theoretical basis for the effective control of M. canis infection and the effect of KTZ on fungi.

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          Most cited references34

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          RNA-Seq: a revolutionary tool for transcriptomics.

          RNA-Seq is a recently developed approach to transcriptome profiling that uses deep-sequencing technologies. Studies using this method have already altered our view of the extent and complexity of eukaryotic transcriptomes. RNA-Seq also provides a far more precise measurement of levels of transcripts and their isoforms than other methods. This article describes the RNA-Seq approach, the challenges associated with its application, and the advances made so far in characterizing several eukaryote transcriptomes.
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            Antifungal agents: mode of action, mechanisms of resistance, and correlation of these mechanisms with bacterial resistance.

            The increased use of antibacterial and antifungal agents in recent years has resulted in the development of resistance to these drugs. The significant clinical implication of resistance has led to heightened interest in the study of antimicrobial resistance from different angles. Areas addressed include mechanisms underlying this resistance, improved methods to detect resistance when it occurs, alternate options for the treatment of infections caused by resistant organisms, and strategies to prevent and control the emergence and spread of resistance. In this review, the mode of action of antifungals and their mechanisms of resistance are discussed. Additionally, an attempt is made to discuss the correlation between fungal and bacterial resistance. Antifungals can be grouped into three classes based on their site of action: azoles, which inhibit the synthesis of ergosterol (the main fungal sterol); polyenes, which interact with fungal membrane sterols physicochemically; and 5-fluorocytosine, which inhibits macromolecular synthesis. Many different types of mechanisms contribute to the development of resistance to antifungals. These mechanisms include alteration in drug target, alteration in sterol biosynthesis, reduction in the intercellular concentration of target enzyme, and overexpression of the antifungal drug target. Although the comparison between the mechanisms of resistance to antifungals and antibacterials is necessarily limited by several factors defined in the review, a correlation between the two exists. For example, modification of enzymes which serve as targets for antimicrobial action and the involvement of membrane pumps in the extrusion of drugs are well characterized in both the eukaryotic and prokaryotic cells.
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              Genomics, gene expression and DNA arrays.

              Experimental genomics in combination with the growing body of sequence information promise to revolutionize the way cells and cellular processes are studied. Information on genomic sequence can be used experimentally with high-density DNA arrays that allow complex mixtures of RNA and DNA to be interrogated in a parallel and quantitative fashion. DNA arrays can be used for many different purposes, most prominently to measure levels of gene expression (messenger RNA abundance) for tens of thousands of genes simultaneously. Measurements of gene expression and other applications of arrays embody much of what is implied by the term 'genomics'; they are broad in scope, large in scale, and take advantage of all available sequence information for experimental design and data interpretation in pursuit of biological understanding.
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                Author and article information

                Journal
                J Vet Sci
                J Vet Sci
                JVS
                Journal of Veterinary Science
                The Korean Society of Veterinary Science
                1229-845X
                1976-555X
                January 2021
                09 December 2020
                : 22
                : 1
                : e4
                Affiliations
                [1 ]Key Laboratory of Livestock Infectious Diseases in Northeast China, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang 110866, China.
                [2 ]Qingdao Vetlab Biotechnology Co., Ltd., Qingdao 266109, China.
                Author notes
                Corresponding author: Yi Zhang. Key Laboratory of Livestock Infectious Diseases in Northeast China, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, 120 Dongling Road, Shenhe District, Shenyang 110866, China. 2005500042@ 123456syau.edu.cn
                Corresponding author: Zeliang Chen. Key Laboratory of Livestock Infectious Diseases in Northeast China, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, 120 Dongling Road, Shenhe District, Shenyang 110866, China. zeliangchen@ 123456yahoo.com

                Mingyang Wang and Yan Zhao contributed equally to this work.

                Author information
                https://orcid.org/0000-0003-3496-3633
                https://orcid.org/0000-0002-2169-2777
                https://orcid.org/0000-0002-7779-1918
                https://orcid.org/0000-0003-0424-1728
                https://orcid.org/0000-0001-5066-9229
                https://orcid.org/0000-0002-7854-1312
                https://orcid.org/0000-0003-4765-184X
                Article
                10.4142/jvs.2021.22.e4
                7850795
                33522156
                da4d5d4f-5ecb-4432-bea6-b113230d1c7b
                © 2021 The Korean Society of Veterinary Science

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

                History
                : 04 September 2020
                : 26 October 2020
                : 05 November 2020
                Funding
                Funded by: Education Department of Liaoning Province
                Award ID: LSNYB201612
                Funded by: National Basic Research Program of China, CrossRef https://doi.org/10.13039/501100012166;
                Award ID: 2017YFD0500901
                Categories
                Original Article
                Microbiology

                Veterinary medicine
                microsporum canis,transcriptome,ketoconazole
                Veterinary medicine
                microsporum canis, transcriptome, ketoconazole

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