Microbiology

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    Protocols in Current Issue
    Rapid in vitro and in vivo Evaluation of Antimicrobial Formulations Using Bioluminescent Pathogenic Bacteria
    Authors:  Artur Schmidtchen and Manoj Puthia, date: 01/20/2022, view: 1958, Q&A: 0
    [Abstract]

    Basic and translational research needs rapid methods to test antimicrobial formulations. Bioluminescent bacteria and advanced imaging systems capable of acquiring bioluminescence enable us to quickly and longitudinally evaluate the efficacy of antimicrobials. Conventional approaches, such as radial diffusion and viable count assays, are

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    Induction of Acute or Disseminating Bacterial Pneumonia in Mice and Sampling of Infected Organs for Studying the Host Response to Bacterial Pneumonia
    Authors:  Wanhai Qin, Zhe Liu, Tom van der Poll and Alex F. de Vos, date: 01/05/2022, view: 2012, Q&A: 0
    [Abstract]

    Experimental pneumonia models are important tools to study the pathophysiology of lung inflammation caused by microbial infections and the efficacy of (novel) drugs. We have applied a murine model of pneumonia induced by Pseudomonas (P.) aeruginosa infection to study acute host antibacterial defense in lungs, and assess epithelial cell specific

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    Selection of Vaccinia Virus Recombinants Using CRISPR/Cas9
    Authors:  Anjali Gowripalan, Stewart A. Smith and David C. Tscharke, date: 12/20/2021, view: 1468, Q&A: 0
    [Abstract]

    The engineering of poxvirus genomes is fundamental to primary and applied virology research. Indeed, recombinant poxviruses form the basis for many novel vaccines and virotherapies but producing and purifying these viruses can be arduous. In recent years, CRISPR/Cas9 has become the favoured approach for genome manipulation due to its speed and

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    A Retro-orbital Sinus Injection Mouse Model to Study Early Events and Reorganization of the Astrocytic Network during Pneumococcal Meningitis
    Authors:  Chakir Bello, Martine Cohen-Salmon and Guy Tran Van Nhieu, date: 12/05/2021, view: 1134, Q&A: 0
    [Abstract]

    Pneumococcal (PN) meningitis is a life-threatening disease with high mortality rates that leads to permanent neurological sequelae. Studies of the process of bacterial crossing of the blood brain barrier (BBB) are hampered by the lack of relevant in vitro and in vivo models of meningitis that recapitulate the human disease. PN meningitis involves

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    Analysis of Protein Stability by Synthesis Shutoff
    Authors:  Ira Buntenbroich, Tânia Simões and Mafalda Escobar-Henriques, date: 11/20/2021, view: 1617, Q&A: 0
    [Abstract]

    In this protocol, we describe the analysis of protein stability over time, using synthesis shutoff. As an example, we express HA-tagged yeast mitofusin Fzo1 in Saccharomyces cerevisiae and inhibit translation via cycloheximide (CHX). Proteasomal inhibition with MG132 is performed, as an optional step, before the addition of CHX. Proteins are

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    Isolation of GFP-expressing Malarial Hypnozoites by Flow Cytometry Cell Sorting
    Authors:  Annemarie Voorberg-van der Wel, Sam O. Hoffman and Clemens H. M. Kocken, date: 05/05/2021, view: 2893, Q&A: 0
    [Abstract]

    Hypnozoites are dormant liver-stage parasites unique to relapsing malarial species, including the important human pathogen Plasmodium vivax, and pose a barrier to the elimination of malaria. Little is known regarding the biology of these stages, largely due to their inaccessible location. Hypnozoites can be cultured in vitro but these cultures

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    A Potent Vaccine Delivery System
    Authors:  Guangzu Zhao, Armira Azuar, Istvan Toth and Mariusz Skwarczynski, date: 04/05/2021, view: 4130, Q&A: 0
    [Abstract]

    Most vaccines require co-delivery of an adjuvant in order to generate the desired immune responses. However, many currently available adjuvants are non-biodegradable, have limited efficacy, and/or poor safety profile. Thus, new adjuvants, or self-adjuvanting vaccine delivery systems, are required. Here, we proposed a self-adjuvanting delivery

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    Resolving Structural Changes of Photoreceptors in Living Escherichia coli via In-cell Infrared Difference Spectroscopy
    Authors:  Lukas Goett-Zink, Jessica L. Klocke and Tilman Kottke, date: 02/05/2021, view: 2931, Q&A: 0
    [Abstract]

    Several in-cell spectroscopic techniques have been developed recently to investigate the structure and mechanism of proteins in their native environment. Conditions in vivo differ dramatically from those selected for in vitro experiments. Accordingly, the cellular environment can affect the protein mechanism for example by molecular crowding or

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    Murine Acute Pneumonia Model of Pseudomonas aeruginosa Lung Infection
    Authors:  Xiaolei Pan and Weihui Wu, date: 11/05/2020, view: 2180, Q&A: 0
    [Abstract] Animal infection models play significant roles in studying bacterial pathogenic mechanisms, host pathogen interaction as well as evaluating drug and vaccine efficacies. We have been utilizing an acute pneumonia model to study bacterial colonization in lungs and assess virulence to the host by determination of bacterial loads and survival assays, ...
    Long-distance Transport in Bacterial Swarms Revealed by Single Nanoparticle Tracking
    Authors:  Jingjing Feng and Yan He, date: 11/05/2020, view: 1696, Q&A: 0
    [Abstract]

    During swarming, high density flagella-driven bacteria migrate collectively in a swirling pattern on wet agar surfaces, immersed in a thin viscous fluid layer called “swarm fluid”. Though the fluid environment has essential role in the emergence of swarming behavior, the microscopic mechanisms of it in mediating the cooperation of

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