Protocols in Current Issue
    TetR Regulated in vivo Repression Technology to Identify Conditional Gene Silencing in Genetically Engineerable Bacteria Using Vibrio cholerae Murine Infections as Model System
    Authors:  Franz G. Zingl, Fabian Mitterer, Himadri B. Thapa and Stefan Schild, date: 10/05/2020, view: 375, Q&A: 0
    [Abstract] Investigation of bacterial gene regulation upon environmental changes is still a challenging task. For example, Vibrio cholerae, a pathogen of the human gastrointestinal tract, faces diverse transient conditions in different compartments upon oral ingestion. Genetic reporter systems have been demonstrated to be extremely powerful tools to ...
    RNA Extraction from Ears and Draining Lymph Nodes of Mice Infected with Leishmania amazonensis
    Authors:  Emilie Giraud and Evie Melanitou, date: 06/05/2020, view: 1136, Q&A: 0
    [Abstract] Parasites of the genus Leishmania infect the mammalian hosts, including mice and humans and cause cutaneous or visceral leishmaniasis depending upon the parasite species transmitted by the vector sandfly. Leishmania amazonensis is one of the Leishmania species responsible for the cutaneous form of the disease. We have ...
    A Triple-challenge Mouse Model of Allergic Airway Disease, Primary Influenza Infection, and Secondary Bacterial Infection
    Authors:  Sean Roberts, Clare M Williams, Sreeja Roy and Yoichi Furuya, date: 04/20/2020, view: 1075, Q&A: 0
    [Abstract] Asthma is a global problem that affects millions of individuals. An increased risk of respiratory viral and bacterial infections is one of the complications of asthma. We recently reported that mice with ovalbumin-induced allergic airway disease (AAD) are protected against influenza-Streptococcus pneumoniae co-infection. Here, we describe ...
    Visualizing Hypoxia in a Murine Model of Candida albicans Infection Using in vivo Biofluorencence
    Authors:  José Pedro Lopes and Constantin F. Urban, date: 08/05/2019, view: 1932, Q&A: 0
    [Abstract] Candida albicans is a leading human fungal pathogen that uses several metabolic adaptations to escape immune cells and causes systemic disease. Here, we describe a protocol for measuring one of these adaptations, the ability to thrive in hypoxic niches. Hypoxia was generated after successful subdermal infection with C. albicans ...
    Use of Gas Chromatography to Quantify Short Chain Fatty Acids in the Serum, Colonic Luminal Content and Feces of Mice
    [Abstract] Short-Chain Fatty Acids (SCFAs) are a product of the fermentation of resistant starches and dietary fibers by the gut microbiota. The most important SCFA are acetate (C2), propionate (C3) and butyrate (C4). These metabolites are formed and absorbed in the colon and then transported through the hepatic vein to the liver. SCFAs are more concentrated ...
    Transplantation of Fecal Microbiota Shaped by Diet
    Authors:  Keiran H. McLeod, Linda Mason and Eliana Mariño, date: 01/05/2018, view: 5728, Q&A: 0
    [Abstract] Alterations in diet and gut microbial ecology underlie the pathogenesis of type 1 diabetes (T1D). In the non-obese diabetic (NOD) mouse, we found high concentrations of bacterial metabolites acetate and butyrate in blood and faeces correlated with protection from disease. We reconstituted germ free (GF) NOD mice with fecal bacteria from protected ...
    Oral Microbiome Characterization in Murine Models
    [Abstract] The oral microbiome has been implicated as a trigger for immune responsiveness in the oral cavity, particularly in the setting of the inflammatory disease periodontitis. The protocol presented here is aimed at characterizing the oral microbiome in murine models at steady state and during perturbations of immunity or physiology. Herein, we describe ...
    Multicolor Stimulated Emission Depletion (STED) Microscopy to Generate High-resolution Images of Respiratory Syncytial Virus Particles and Infected Cells
    [Abstract] Human respiratory syncytial virus (RSV) infection in human lung epithelial A549 cells induces filopodia, cellular protrusions consisting of F-actin, that extend to neighboring uninfected cells (Mehedi et al., 2016). High-resolution imaging via stimulated emission depletion (STED) microscopy revealed filamentous RSV particles along these ...
    Infection of Human Hepatocyte-chimeric Mice with HBV and in vivo Treatment with εRNA
    Authors:  Seiichi Sato, Kai Li and Akinori Takaoka, date: 01/20/2016, view: 5540, Q&A: 0
    [Abstract] Hepatitis B virus (HBV) can cause both acute and chronic disease in human liver with potentially high risk of cirrhosis and liver cancer. The host range of non-human primates susceptible to this virus is limited. Therefore, experimental studies with human hepatocyte-chimeric mice provide an invaluable source of information regarding the biology ...
    TCRβ Clonotype Analysis of EBV and CMV-specific Human CD8+ T Cells
    Authors:  Nening M. Nanlohy, Dan Koning, Esther D. Quakkelaar and Debbie van Baarle, date: 10/05/2015, view: 5700, Q&A: 0
    [Abstract] This protocol describes the quantification of all expressed T-cell antigen receptor (TCR) gene products within sorted (by flow cytometry) EBV and CMV-specific memory CD8+ T-cell populations using a template-switch anchored reverse transcription polymerase chain reaction (RT-PCR).

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