Microbiology

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    Protocols in Current Issue
    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: 2053, 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: 1435, 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: 3218, 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: 2468, 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 ...
    Nab Escaping AAV Mutants Isolated from Mouse Muscles
    Authors:  Zheng Chai, R. Jude Samulski and Chengwen Li, date: 05/05/2018, view: 2446, Q&A: 0
    [Abstract] Neutralizing antibodies (Nabs) are a major challenge in clinical trials of adeno-associated virus (AAV) vector gene therapy, because Nabs are able to inhibit AAV transduction in patients. We have successfully isolated several novel Nab-escaped AAV chimeric capsids in mice by administrating a mixture of AAV shuffled library and patient serum. These ...
    Identification and Quantification of Secondary Metabolites by LC-MS from Plant-associated Pseudomonas aurantiaca and Pseudomonas chlororaphis
    Authors:  Izzah Shahid, Muhammad Rizwan and Samina Mehnaz, date: 01/20/2018, view: 4364, Q&A: 0
    [Abstract] Increased antibiotic resistance of plants and human pathogens and continuous use of chemical fertilizers has pushed microbiologists to explore new microbial sources as potential antagonists. In this study, eight strains of Pseudomonas aurantiaca and Pseudomonas chlororaphis, have been isolated from different plant sources and ...
    Bacterial Aggregation Assay in the Presence of Cyclic Lipopeptides
    Authors:  Pengyuan Xiu, Rui Liu, Dechao Zhang and Chaomin Sun, date: 01/05/2018, view: 3176, Q&A: 0
    [Abstract] Lipopeptides is an important class of biosurfactants having antimicrobial and anti-adhesive activity against pathogenic bacteria. These include surfactin, fengycin, iturin, bacillomycin, mycosubtilin, lichenysin, and pumilacidin (Arima et al., 1968; Naruse et al., 1990; Yakimov et al., 1995; Steller and Vater, 2000; ...
    Quantification of Trypanosoma cruzi in Tissue and Trypanosoma cruzi Killing Assay
    Authors:  Hisako Kayama, Shoko Kitada and Kiyoshi Takeda, date: 11/20/2017, view: 3636, Q&A: 0
    [Abstract] Infection with Trypanosoma cruzi causes Chagas disease. The methods provided here allow for the quantification of T. cruzi in the liver, heart, and blood of intraperitoneally-infected mice and analysis of the killing activity of the cells infected with T. cruzi in vitro.
    In vitro Assay to Assess Efficacy of Potential Antiviral Compounds against Enterovirus D68
    Authors:  Liang Sun, Leen Delang, Carmen Mirabelli and Johan Neyts, date: 03/20/2017, view: 4784, Q&A: 0
    [Abstract] In 2014 enterovirus D68 (EV-D68) caused the largest outbreak in the United States since the discovery of the virus. Distinct from before, the 2014 infections were associated with more severe respiratory disease and occasional neurological complications. So far, there are no available vaccines or antivirals for the prophylaxis or treatment of ...
    Antibiotic Disc Assay for Synechocystis sp. PCC6803
    Authors:  Otilia Cheregi and Christiane Funk, date: 12/20/2016, view: 5550, Q&A: 0
    [Abstract] This protocol describes how to investigate the integrity of the outer cell wall in the cyanobacterium Synechocystis sp. PCC6803 using antibiotics. It is adapted to the agar diffusion test (Bauer et al., 1966), in which filter paper discs impregnated with specified concentrations of antibiotics were placed on agar plates ...



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