Fungi

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    Protocols in Current Issue
    ODELAM: Rapid Sequence-independent Detection of Drug Resistance in Mycobacterium tuberculosis Isolates
    Authors:  Thurston Herricks, Magdalena Donczew, David R. Sherman and John D. Aitchison, date: 05/20/2021, view: 390, Q&A: 0
    [Abstract]

    Antimicrobial-resistant Mycobacterium tuberculosis (Mtb) causes over 200,000 deaths globally each year. Current assays of antimicrobial resistance require knowledge of the mutations that confer drug resistance or long periods of culture time to test growth under drug pressure. We present ODELAM (One-cell Doubling Evaluation of Living Arrays of

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    Chromatographic Assays for the Enzymatic Degradation of Chitin
    Authors:  Sophanit Mekasha, Tina R. Tuveng, Gustav Vaaje-Kolstad and Vincent G. H. Eijsink, date: 05/05/2021, view: 506, Q&A: 0
    [Abstract]

    Chitin is an insoluble linear polymer of β(1→4)-linked N-acetylglucosamine. Enzymatic cleavage of chitin chains can be achieved using hydrolytic enzymes, called chitinases, and/or oxidative enzymes, called lytic polysaccharide monooxygenases (LPMOs). These two groups of enzymes have different modes of action and yield different product

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    Inkjet 3D Printing of Polymers Resistant to Fungal Attachment
    Authors:  Yinfeng He, Cindy Vallieres, Morgan R. Alexander, Ricky D. Wildman and Simon V. Avery, date: 05/05/2021, view: 605, Q&A: 0
    [Abstract]

    Inkjet 3D printing is an additive manufacturing method that allows the user to produce a small batch of customized devices for comparative study versus commercial products. Here, we describe the use of a commercial 2D ink development system (Dimatix material printing) to manufacture small batches of 3D medical or other devices using a recently

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    Single Cell Analysis and Sorting of Aspergillus fumigatus by Flow Cytometry
    Authors:  Gareth Howell and Robert-Jan Bleichrodt, date: 04/20/2021, view: 825, Q&A: 0
    [Abstract]

    Experimental results in fungal biology research are usually obtained as average measurements across whole populations of cells, whilst ignoring what is happening at the single cell level. Microscopy has allowed us to study single-cell behavior, but it has low throughput and cannot be used to select individual cells for downstream experiments. Here

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    Quantitative Characterization of the Amount and Length of (1,3)-β-D-glucan for Functional and Mechanistic Analysis of Fungal (1,3)-β-D-glucan Synthase
    Authors:  Abhishek Chhetri, Anna Loksztejn and Kenichi Yokoyama, date: 04/20/2021, view: 967, Q&A: 0
    [Abstract]

    (1,3)-β-d-Glucan synthase (GS) is an essential enzyme for fungal cell wall biosynthesis that catalyzes the synthesis of (1,3)-β-d-glucan, a major and vital component of the cell wall. GS is a proven target of antifungal antibiotics including FDA-approved echinocandin derivatives; however, the function and mechanism of GS remain largely

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    Analysis of TORC1-body Formation in Budding Yeast
    Authors:  Ryan L. Wallace, Eric Lu, Arron Sullivan, James E. Hughes Hallett and Andrew P. Capaldi, date: 04/05/2021, view: 499, Q&A: 0
    [Abstract]

    The Target of Rapamycin kinase Complex I (TORC1) is the master regulator of cell growth and metabolism in eukaryotes. In the presence of pro-growth hormones and abundant nutrients, TORC1 is active and drives protein, lipid, and nucleotide synthesis by phosphorylating a wide range of proteins. In contrast, when nitrogen and/or glucose levels fall,

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    Transcardiac Perfusion of the Mouse for Brain Tissue Dissection and Fixation
    Authors:  Jinyun Wu, Yuqi Cai, Xiaoyang Wu, Yue Ying, Yilin Tai and Miao He, date: 03/05/2021, view: 1343, Q&A: 0
    [Abstract] Transcardiac perfusion with saline followed by 4% paraformaldehyde (PFA) is widely used to clear blood and preserve brain for immunostaining or in situ hybridization. PFA breaks into formaldehyde in solution, which cross-link protein and DNA molecules to preserve tissue and cell structure. Here we provide a step by step guide for ...
    Candida Biofilm Formation Assay on Essential Oil Coated Silicone Rubber
    [Abstract]

    Development of biofilm associated candidemia for patients with implanted biomaterials causes an urgency to develop antimicrobial and biofilm inhibitive coatings in the management of recalcitrant Candida infections. Recently, there is an increase in the number of patients with biofilm formation and resistance to antifungal therapy. Therefore, there

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    EmPC-seq: Accurate RNA-sequencing and Bioinformatics Platform to Map RNA Polymerases and Remove Background Error
    [Abstract]

    Transcription errors can substantially affect metabolic processes in organisms by altering the epigenome and causing misincorporations in mRNA, which is translated into aberrant mutant proteins. Moreover, within eukaryotic genomes there are specific Transcription Error-Enriched genomic Loci (TEELs) which are transcribed by RNA polymerases with

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    Polyamine Transport Assay Using Reconstituted Yeast Membranes
    Authors:  Sarah van Veen, Shaun Martin, Marleen Schuermans and Peter Vangheluwe, date: 01/20/2021, view: 1226, Q&A: 0
    [Abstract]

    ATP13A2/PARK9 is a late endo-/lysosomal P5B transport ATPase that is associated with several neurodegenerative disorders. We recently characterized ATP13A2 as a lysosomal polyamine exporter, which sheds light on the molecular identity of the unknown mammalian polyamine transport system. Here, we describe step by step a protocol to measure

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