Protocols in Current Issue
    Fluorescence-based Heme Quantitation in Toxoplasma Gondii
    Authors:  Amy Bergmann and Zhicheng Dou, date: 06/20/2021, view: 886, Q&A: 0

    Toxoplasma gondii is a highly prevalent protozoan pathogen throughout the world. As a eukaryotic intracellular pathogen, Toxoplasma ingests nutrients from host cells to support its intracellular growth. The parasites also encode full or partial metabolic pathways for the biosynthesis of certain nutrients, such as heme. Heme is an essential

    Sex-specific Separation of Plasmodium falciparum Gametocyte Populations
    Authors:  Melanie C. Ridgway, Daniela Cihalova and Alexander G. Maier, date: 06/05/2021, view: 1089, Q&A: 0

    Plasmodium falciparum is a unicellular eukaryotic parasite that causes malaria in humans. The parasite is spread by Anopheles mosquitoes after ingestion of sexual stage parasites known as gametocytes. Malaria transmission depends on parasites switching from the disease-causing asexual blood forms to male and female gametocytes. The current

    Preparation of Doublet Microtubule Fraction for Single Particle Cryo-electron Microscopy
    Authors:  Corbin Black, Daniel Chen Dai, Katya Peri, Muneyoshi Ichikawa and Khanh Huy Bui, date: 06/05/2021, view: 1578, Q&A: 0

    Over the years, studying the ultrastructure of the eukaryotic cilia/flagella using electron microscopy (EM) has contributed significantly toward our understanding of ciliary function. Major complexes in the cilia, such as inner and outer dynein arms, radial spokes, and dynein regulatory complexes, were originally discovered by EM. Classical

    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: 1401, Q&A: 0

    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

    Click Chemistry for Imaging in-situ Protein Palmitoylation during the Asexual Stages of Plasmodium falciparum
    Author:  Mansoor A. Siddiqui, date: 05/05/2021, view: 1898, Q&A: 0

    Palmitoylation refers to the modification of the cysteine thiols in proteins by fatty acids, most commonly palmitic acid, through ‘thioester bond’ formation. In vivo, palmitoylation of proteins is catalyzed by palmitoyl acyltransferases (PATs or DHHC-PATs). Palmitoylation has recently emerged as a crucial post-translational

    Trypanosomatid, fluorescence-based in vitro U-insertion/U-deletion RNA-editing (FIDE)
    Authors:  Wolf-Matthias Leeder, Elisabeth Kruse and H. Ulrich Göringer, date: 03/05/2021, view: 1651, Q&A: 0

    Gene expression within the mitochondria of African trypanosomes and other protozoan organisms relies on a nucleotide-specific RNA-editing reaction. In the process exclusively uridine (U)-nucleotides are site-specifically inserted into and deleted from sequence-deficient primary transcripts to convert them into translatable mRNAs. The reaction is

    Plant ARGONAUTE Protein Immunopurification for Pathogen Cross Kingdom Small RNA Analysis
    Authors:  Florian Dunker, Bernhard Lederer and Arne Weiberg, date: 02/05/2021, view: 1894, Q&A: 0

    Over the last decade, it has been noticed that microbial pathogens and pests deliver small RNA (sRNA) effectors into their host plants to manipulate plant physiology and immunity for infection, known as cross kingdom RNA interference. In this process, fungal and oomycete parasite sRNAs hijack the plant ARGONAUTE (AGO)/RNA-induced silencing complex

    Determination of Intracellular Ca2+ Concentration in the Human Pathogens Trypanosomatids Leishmania mexicana and Trypanosoma cruzi by the Use of the Fluorescent Ca2+ Indicator Fura-2
    Authors:  Andre Rey-Cibati, Mariana Valladares-Delgado and Gustavo Benaim, date: 09/20/2020, view: 1596, Q&A: 0
    [Abstract] Ca2+ is an essential signaling messenger in all eukariotic cells, playing a pivotal role in many cellular functions as cell growth control (differentiation, fertilization and apoptosis), secretion, gene expression, enzyme regulation, among many others. This basic premise includes trypanosomatids as Trypanosoma cruzi and various ...
    An in vitro DNA Sensor-based Assay to Measure Receptor-specific Adhesion Forces of Eukaryotic Cells and Pathogens
    Authors:  Maurizio Wack, Tina Wiegand, Friedrich Frischknecht and E. Ada Cavalcanti-Adam, date: 09/05/2020, view: 1858, Q&A: 0
    [Abstract] Motility of eukaryotic cells or pathogens within tissues is mediated by the turnover of specific interactions with other cells or with the extracellular matrix. Biophysical characterization of these ligand-receptor adhesions helps to unravel the molecular mechanisms driving migration. Traction force microscopy or optical tweezers are typically ...
    Multiplication and Growth Inhibition Activity Assays for the Zoonotic Malaria Parasite, Plasmodium knowlesi
    Authors:  Franziska Mohring, Thomas A. Rawlinson, Simon J. Draper and Robert W. Moon, date: 09/05/2020, view: 2473, Q&A: 0
    [Abstract] Malaria remains a major cause of morbidity and mortality globally. Clinical symptoms of the disease arise from the growth and multiplication of Plasmodium parasites within the blood of the host. Thus in vitro assays to determine how drug, antibody and genetic perturbations affect the growth rate of Plasmodium parasites ...

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