Protocols in Current Issue
    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: 283, 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: 913, 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: 621, 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 ...
    In vitro Cultivation and Visualization of Malaria Liver Stages in Primary Simian Hepatocytes
    Authors:  Devendra Kumar Gupta and Thierry Diagana, date: 08/20/2020, view: 701, Q&A: 0
    [Abstract] Human liver is the primary and obligatory site for malaria infection where sporozoites invade host hepatocytes. Malaria hepatic stages are asymptomatic and represent an attractive target for development of anti-malarial interventions and vaccines. However, owing to lack of robust and reproducible in vitro culture system, it is difficult ...
    Microscopy-based Methods for Rosetting Assay in Malaria Research
    Authors:  Wenn-Chyau Lee and Laurent Rénia, date: 07/05/2020, view: 1015, Q&A: 0
    [Abstract] In malaria, rosetting phenomenon is a condition where a Plasmodium-infected erythrocyte stably adheres to at least an uninfected erythrocyte. This phenomenon that occurs in all species of human malaria parasite is likely to be an immune escape mechanism for the parasite. However, it has been associated with malaria pathogenesis, possibly ...
    Differential Fractionation of Erythrocytes Infected by Plasmodium berghei
    Authors:  Bénédicte Gnangnon, Véronique Peucelle and Christine Pierrot, date: 06/05/2020, view: 727, Q&A: 0
    [Abstract] The study of host/pathogen interactions at the cellular level during Plasmodium intra-erythrocytic cycle requires differential extraction techniques aiming to analyze the different compartments of the infected cell. Various protocols have been proposed in the literature to study specific compartments and/or membranes in the infected ...
    Viral Double-Stranded RNA Detection by DNase I and Nuclease S1 digestions in Leishmania parasites
    Authors:  Nathalie Isorce and Nicolas Fasel, date: 05/05/2020, view: 806, Q&A: 0
    [Abstract] Many RNA viruses are found in protozoan parasites. They can be responsible for more serious pathology or treatment failure. For the detection of viral double-stranded RNA (dsRNA), sequence-dependent and -independent methods are available, such as quantitative real-time PCR and immunofluorescence, dot blot, ELISA or sequencing. The technique ...
    Spectrophotometric Assessment of Heme Oxygenase-1 Activity in Leishmania-infected Macrophages
    Authors:  Moumita Basu, Shriya Saha and Anindita Ukil, date: 04/05/2020, view: 1078, Q&A: 0
    [Abstract] Heme oxygenase-1 (HO-1) is a stress responsive enzyme that metabolizes heme and releases free iron, carbon monoxide (CO), and biliverdin (BV), which rapidly undergoes conversion to bilirubin (BL). Estimation of bilirubin is the basis of HO-1 assay. HO-1 activity is widely employed to determine antioxidant response of cells under different ...
    CRISPR-Cas9 Genome Editing of Plasmodium knowlesi
    Authors:  Franziska Mohring, Melissa N. Hart, Avnish Patel, David A. Baker and Robert W. Moon, date: 02/20/2020, view: 1591, Q&A: 1
    [Abstract] Plasmodium knowlesi is a zoonotic malaria parasite in Southeast Asia that can cause severe and fatal malaria in humans. The main hosts are Macaques, but modern diagnostic tools reveal increasing numbers of human infections. After P. falciparum, P. knowlesi is the only other malaria parasite capable of being maintained in ...
    Surface Plasmon Resonance Analysis of the Protein-protein Binding Specificity Using Autolab ESPIRIT
    Authors:  Pragyan Parimita Rath, Gaurav Anand and Shalini Agarwal, date: 02/20/2020, view: 1977, Q&A: 0
    [Abstract] Direct protein-protein interactions are known to regulate a wide range of cellular activities. To understand these contacts one can employ various experimental methods like Dynamic Light Scattering (DLS), Fluorescence Resonance Energy Transfer (FRET), Isothermal titration calorimetry (ITC), Chemical crosslinking, Co-immunoprecipitation (Co-IP), ...

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