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Live-cell Imaging by Super-resolution Confocal Live Imaging Microscopy (SCLIM): Simultaneous Three-color and Four-dimensional Live Cell Imaging with High Space and Time Resolution

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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: 694, 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: 439, 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: 573, 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: 877, 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: 596, 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 ...

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: 1420, Q&A: 0

[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 ...

MicroScale Thermophoresis as a Tool to Study Protein-peptide Interactions in the Context of Large Eukaryotic Protein Complexes

Authors: Maximilian G. Plach, Klaus Grasser and Thomas Schubert, date: 12/05/2017, view: 14645, Q&A: 0

[Abstract] Protein-peptide interactions are part of many physiological processes, for example, epigenetics where peptide regions of histone complexes are crucial for regulation of chromatin structure. Short peptides are often also used as alternatives to small molecule drugs to target protein complexes. Studying the interactions between proteins and peptides ...

Liposome Disruption Assay to Examine Lytic Properties of Biomolecules

Authors: John R. Jimah, Paul H. Schlesinger and Niraj H. Tolia, date: 08/05/2017, view: 8320, Q&A: 0

[Abstract] Proteins may have three dimensional structural or amino acid features that suggest a role in targeting and disrupting lipids within cell membranes. It is often necessary to experimentally investigate if these proteins and biomolecules are able to disrupt membranes in order to conclusively characterize the function of these biomolecules. Here, we ...

Plasmodium Sporozoite Motility on Flat Substrates

Authors: Henriette L Prinz, Julia M Sattler and Friedrich Frischknecht, date: 07/20/2017, view: 5901, Q&A: 0

[Abstract] Plasmodium sporozoites are the infectious, highly motile forms of the malaria parasite transmitted by Anopheles mosquitoes. Sporozoite motility can be assessed following the dissection of Anopheles salivary glands and isolation of sporozoites in vitro.

A Gas Chromatography-Mass Spectrometry-Based Two Stage Assay for Measurement of in vitro myo-Inositol 3-phosphate Synthase (INO1) Activity

Authors: James I. MacRae and Malcolm J. McConville, date: 03/05/2015, view: 7547, Q&A: 0

[Abstract] This method describes an in vitro assay for measuring INO1 enzyme activity (the conversion of glucose 6-phosphate to myo-inositol 3-phosphate) in cell-free extracts. The method was first described for Plasmodium falciparum cells in MacRae et al. (2014) and consists of two parts: Part 1 describes the assay itself ...

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