Protocols in Current Issue
    Neutrophil Extracellular Trap Killing Assay of Candida albicans
    Authors:  Sheng-Yang Wu and Betty A. Wu-Hsieh, date: 08/20/2020, view: 721, Q&A: 0
    [Abstract] Fungal pathogen Candida albicans is one of the top leading causes of overall healthcare-associated bloodstream infections worldwide. Neutrophil is the major effector cell to clear C. albicans infection. Our study showed that mouse neutrophils utilize two independent mechanisms to kill C. albicans: one is CR3 downstream ...
    Contemporaneous Measurement of Outer and Inner Membrane Permeability in Gram-negative Bacteria
    Authors:  Bo Ma, Chao Fang, Jing Zhang, Mingzhi Wang, Xiaoxing Luo and Zheng Hou, date: 03/05/2020, view: 1132, Q&A: 0
    [Abstract] The emergence and rapid spread of multidrug resistance in bacteria have led to the urgent need for novel antibacterial agents. Membrane permeabilization is the mechanism for many antibacterial molecules that are being developed against gram-negative bacteria. Thus, to determine the efficacy of a potential antibacterial molecule, it is important to ...
    Rapid Detection of Proliferative Bacteria by Electrical Stimulation
    Authors:  Conor LA Edwards, Dmitry Malyshev, James P Stratford and Munehiro Asally, date: 02/05/2020, view: 2013, Q&A: 0
    [Abstract] Detecting live bacteria is an important task for antimicrobial susceptibility testing (AST) in the medical sector and for quality-monitoring in biological industries. Current methods for live-bacteria detection suffer limitations in speed or sensitivity. In a recent paper, we reported that electrical response dynamics in membrane potential enable ...
    Competition Assays to Quantify the Effect of Biocontrol Yeasts against Plant Pathogenic Fungi on Fruits
    Authors:  Electine Magoye, Melanie Pfister, Maja Hilber-Bodmer and Florian M. Freimoser, date: 02/05/2020, view: 1099, Q&A: 0
    [Abstract] Yeasts such as Aureobasidium pullulans are unicellular fungi that occur in all environments and play important roles in biotechnology, medicine, food and beverage production, research, and agriculture. In the latter, yeasts are explored as biocontrol agents for the control of plant pathogenic fungi (e.g., Botrytis cinerea, ...
    A Radioactive-free Kinase Inhibitor Discovery Assay Against the Trypanosoma brucei Glycogen Synthase Kinase-3 short (TbGSK-3s)
    Authors:  Antonia Efstathiou and Despina Smirlis, date: 01/20/2020, view: 986, Q&A: 0
    [Abstract] The identification of small molecules possessing inhibitory activity in vitro, against a given target kinase, is the first step in the drug discovery process. Herein, we describe a non radioactive protocol using luciferase-based ATP assay for the identification of inhibitors for the short isoform of the Trypanosoma brucei’s ...
    In vitro Screening of Antileishmanial Activity of Natural Product Compounds: Determination of IC50, CC50 and SI Values
    Authors:  Olga S. Koutsoni, Kalliopi Karampetsou and Eleni Dotsika, date: 11/05/2019, view: 1465, Q&A: 0
    [Abstract] Neglected tropical diseases gain the scientific interest of numerous research programs in an attempt to achieve their effective control or elimination. In this attempt, more cutting-edge public health policies and research are needed for the discovery of new, safer and effective drugs originated from natural products. Here, we describe protocols ...
    Assessing Membrane Fluidity and Visualizing Fluid Membrane Domains in Bacteria Using Fluorescent Membrane Dyes
    Authors:  Michaela Wenzel, Norbert O. E. Vischer, Henrik Strahl and Leendert W. Hamoen, date: 10/20/2018, view: 4448, Q&A: 0
    [Abstract] Membrane fluidity is a key parameter of bacterial membranes that undergoes quick adaptation in response to environmental challenges and has recently emerged as an important factor in the antibacterial mechanism of membrane-targeting antibiotics. The specific level of membrane fluidity is not uniform across the bacterial cell membrane. Rather, ...
    Analysis of the Effect of Sphingomyelinase on Rubella Virus Infectivity in Two Cell Lines
    Authors:  Noriyuki Otsuki, Masafumi Sakata, Yoshio Mori, Kiyoko Okamoto and Makoto Takeda, date: 09/05/2018, view: 2855, Q&A: 0
    [Abstract] Rubella is a mildly contagious disease characterized by low-grade fever and a morbilliform rash caused by the rubella virus (RuV). Viruses often use cellular phospholipids for infection. We studied the roles of cellular sphingomyelin in RuV infection. Treatment of cells with sphingomyelinase (SMase) inhibited RuV infection in rabbit kidney-derived ...
    Virucidal and Neutralizing Activity Tests for Antiviral Substances and Antibodies
    Authors:  Chie Aoki-Utsubo, Ming Chen and Hak Hotta, date: 05/20/2018, view: 6925, Q&A: 0
    [Abstract] In a narrow definition, virucidal activity represents the activity by which to interact with and physically disrupt viral particles. In a broad definition, it includes the activity by which to functionally inhibit (neutralize) viral infectivity without apparent morphological alterations of the viral particles. The viral infectivity can be measured ...
    Time-of-addition and Temperature-shift Assays to Determine Particular Step(s) in the Viral Life Cycle that is Blocked by Antiviral Substance(s)
    Authors:  Chie Aoki-Utsubo, Ming Chen and Hak Hotta, date: 05/05/2018, view: 4634, Q&A: 0
    [Abstract] Viruses infect their host cells to produce progeny virus particles through the sequential steps of the viral life cycle, such as viral attachment, entry, penetration and post-entry events. This protocol describes time-of-addition and temperature-shift assays that are employed to explore which step(s) in the viral life cycle is blocked by an ...

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