Microbiology

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    Protocols in Current Issue
    Model of Chemotherapy-associated Mucositis and Oral Opportunistic Infections
    Authors:  Takanori Sobue, Martinna Bertolini, Angela Thompson and Anna Dongari-Bagtzoglou, date: 11/05/2019, view: 104, Q&A: 0
    [Abstract] Oral mucositis is a common complication of cancer chemotherapy treatment. Because of the lack of relevant oral mucositis experimental models, it is not clear how chemotherapeutic agents injure the oral mucosa and if commensal microorganisms accelerate tissue damage. We developed an organotypic oral mucosa model that mimics cellular responses ...
    Intracellular Invasion and Killing Assay to Investigate the Effects of Binge Alcohol Toxicity in Murine Alveolar Macrophages
    Authors:  Victor Jimenez Jr and Fernando P Monroy, date: 01/20/2019, view: 1431, Q&A: 0
    [Abstract] Alcohol consumption has diverse and well-documented effects on the human immune system and its ability to defend against infective agents. While pulmonary related infections can occur in healthy humans, binge alcohol use is recognized as a major health risk factor (Nelson et al., 1991). Although binge alcohol consumption has been ...
    Sleeping Beauty Transposon-based System for Rapid Generation of HBV-replicating Stable Cell Lines
    Authors:  Jin-Wei Zheng, Jia-Li Cao and Quan Yuan , date: 07/05/2018, view: 2893, Q&A: 0
    [Abstract] The stable HBV-transfected cell lines, which based on stable integration of replication-competent HBV genome into hepatic cells, are widely used in basic research and antiviral drug evaluation against HBV. However, previous reported strategies to generate HBV-replicating cell lines, which primarily rely on random integration of exogenous DNA by ...
    Adhesion of Enteroaggregative E. coli Strains to HEK293 Cells
    Authors:  Jorge Luis Ayala-Lujan and Fernando Ruiz-Perez, date: 04/20/2018, view: 2755, Q&A: 0
    [Abstract] Enteroaggregative Escherichia coli (EAEC) is a recognized cause of acute diarrhea among both children and adults worldwide. EAEC strains are characterized by the presence of aggregative adherence fimbriae (AAF), which play a key role in pathogenesis by mediating attachment to the intestinal mucosa and by triggering host inflammatory ...
    Quantification of Colibactin-associated Genotoxicity in HeLa Cells by In Cell Western (ICW) Using γ-H2AX as a Marker
    Authors:  Sophie Tronnet and Eric Oswald, date: 03/20/2018, view: 2902, Q&A: 0
    [Abstract] The genotoxin colibactin is produced by several species of Enterobacteriaceae. This genotoxin induces DNA damage, cell cycle arrest, senescence and death in eukaryotic cells (Nougayrède et al., 2006; Taieb et al., 2016). Here we describe a method to quantify the genotoxicity of bacteria producing colibactin following a ...
    Adhesion and Invasion Assay Procedure Using Caco-2 Cells for Listeria monocytogenes
    Authors:  Swetha Reddy and Frank Austin, date: 05/05/2017, view: 7729, Q&A: 0
    [Abstract] Listeria monocytogenes is an important Gram-positive foodborne pathogen that is a particular problem in ready-to-eat food. It has an ability to survive in harsh conditions like refrigeration temperatures and high salt concentrations and is known to cross intestinal, placental and blood-brain barriers. Several cancerous cell lines like ...
    Isolation of Exosomes from Semen for in vitro Uptake and HIV-1 Infection Assays
    Authors:  Marisa N. Madison, Jennifer L. Welch and Chioma M. Okeoma, date: 04/05/2017, view: 6075, Q&A: 0
    [Abstract] Exosomes are membranous extracellular nanovesicles of endocytic origin. Exosomes are known to carry host and pathogen-derived genomic, proteomic, lipidomic cargos and other extraneous molecules. Exosomes are secreted by diverse cell types into the extracellular milieu and are subsequently internalized by recipient neighboring or distal cells. Upon ...
    Transient Transfection-based Fusion Assay for Viral Proteins
    Authors:  Melina Vallbracht, Christina Schröter, Barbara G. Klupp and Thomas C. Mettenleiter, date: 03/05/2017, view: 6040, Q&A: 1
    [Abstract] Membrane fusion is vital for entry of enveloped viruses into host cells as well as for direct viral cell-to-cell spread. To understand the fusion mechanism in more detail, we use an infection free system whereby fusion can be induced by a minimal set of the alphaherpesvirus pseudorabies virus (PrV) glycoproteins gB, gH and gL. Here, we describe an ...
    Murine Leukemia Virus (MLV)-based Coronavirus Spike-pseudotyped Particle Production and Infection
    Authors:  Jean Kaoru Millet and Gary R. Whittaker, date: 12/05/2016, view: 5686, Q&A: 0
    [Abstract] Viral pseudotyped particles (pp) are enveloped virus particles, typically derived from retroviruses or rhabdoviruses, that harbor heterologous envelope glycoproteins on their surface and a genome lacking essential genes. These synthetic viral particles are safer surrogates of native viruses and acquire the tropism and host entry pathway ...
    Purification of Bacterial RNA from Infected Macrophages
    Authors:  Lior Lobel, Nadejda Sigal, Anna Pasechnek and Anat A. Herskovits, date: 11/20/2015, view: 6586, Q&A: 0
    [Abstract] Studying the transcriptome of bacterial pathogens during infection is a very informative and effective tool for discovering genes that contribute to successful infection. However, isolating bacterial RNA from infected cells or tissues is a challenging process due to the much higher amounts of host RNA in the lysates of infected cells. We have ...



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