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    Protocols in Current Issue
    Multiplex T-cell Stimulation Assay Utilizing a T-cell Activation Reporter-based Detection System
    Authors:  Laurie G. Landry, Sarah E. Mann, Amanda M. Anderson and Maki Nakayama, date: 01/20/2021, view: 29, Q&A: 0

    Immune tolerance and response are both largely driven by the interactions between the major histocompatibility complex (MHC) expressed by antigen presenting cells (APCs), T-cell receptors (TCRs) on T-cells, and their cognate antigens. Disordered

    ...
    Wounding Caenorhabditis elegans with Glass Wool

    Research on wound healing majorly relies on rat, mice and other animal models. However, an alternative animal model ought to be brought in the field, pertaining to the stringent ethical issues owing to the use of animals in research. In this regard,

    ...
    Determination of Antibacterial Activity of Film Coatings against Four Clinically Relevant Bacterial Strains

    Antibacterial coatings have currently gained great importance in biomedical technology investigations. Because of the spatial arrangement of the film coatings, evaluation of antibacterial activity presents a new challenge regarding traditional

    ...
    Polyamine Transport Assay Using Reconstituted Yeast Membranes
    Authors:  Sarah van Veen, Shaun Martin, Marleen Schuermans and Peter Vangheluwe, date: 01/20/2021, view: 17, Q&A: 0

    ATP13A2/PARK9 is a late endo-/lysosomal P5B transport ATPase that is associated with several neurodegenerative disorders. We recently characterized ATP13A2 as a lysosomal polyamine exporter, which sheds light on the molecular identity of the unknown

    ...
    Imaging of Human Cancer Cells in 3D Collagen Matrices
    Authors:  Karin Pfisterer, Brooke Lumicisi and Maddy Parsons, date: 01/20/2021, view: 18, Q&A: 0

    Research on cell migration and interactions with the extracellular matrix (ECM) was mostly focused on 2D surfaces in the past. Many recent studies have highlighted differences in migratory behaviour of cells on 2D surfaces compared to complex cell

    ...
    Histological and Immunohistochemical Examination of Stem Cell Proliferation and Reepithelialization in the Wounded Skin
    Authors:  Rupali Gund, Ravindra Zirmire, Haarshaadri J., Gaurav Kansagara and Colin Jamora, date: 01/20/2021, view: 19, Q&A: 0

    The skin is the largest organ that protects our body from the external environment and it is constantly exposed to pathogenic insults and injury. Repair of damage to this organ is carried out by a complex process involving three overlapping phases

    ...
    Evaluation of Toxicity with Brine Shrimp Assay
    Authors:  Christina N. Banti and Sotiris K. Hadjikakou, date: 01/20/2021, view: 20, Q&A: 0

    The in vivo toxicity of new metallodrugs either as Small Bioactive Molecules (SBAMs) or Conjugates of Metals with Drugs (CoMeDs) or their hydrogels such as with hydroxyethyl-methacrylate (HEMA) (pHEMA@SBAMs or pHEMA@CoMeDs) are evaluated by the

    ...
    Preparation of Synaptoneurosomes to Study the Synapse in the Murine Cerebral Cortex
    Authors:  Ariel Diaz, Enrique Torre and Manuel Yepes, date: 01/20/2021, view: 16, Q&A: 0

    The synapse is a complex structure where the transmission of information takes place. Synaptic dysfunction is one of the earliest pathophysiological events in several diseases, such as traumatic brain injury, cerebral ischemia, and neurodegenerative

    ...
    Preparation of Synaptoneurosomes to Study the Synapse in the Murine Cerebral Cortex
    Authors:  Ariel Diaz, Enrique Torre and Manuel Yepes, date: 01/20/2021, view: 16, Q&A: 0
    [Abstract]

    The synapse is a complex structure where the transmission of information takes place. Synaptic dysfunction is one of the earliest pathophysiological events in several diseases, such as traumatic brain injury, cerebral ischemia, and neurodegenerative diseases. Thus, a methodology to study synaptic structure and function is crucial for the

    ...
    Evaluation of Toxicity with Brine Shrimp Assay
    Authors:  Christina N. Banti and Sotiris K. Hadjikakou, date: 01/20/2021, view: 20, Q&A: 0
    [Abstract]

    The in vivo toxicity of new metallodrugs either as Small Bioactive Molecules (SBAMs) or Conjugates of Metals with Drugs (CoMeDs) or their hydrogels such as with hydroxyethyl-methacrylate (HEMA) (pHEMA@SBAMs or pHEMA@CoMeDs) are evaluated by the brine shrimp assay. Thus individuals of Artemia salina larvae are incubated in saline solutions with

    ...
    Histological and Immunohistochemical Examination of Stem Cell Proliferation and Reepithelialization in the Wounded Skin
    Authors:  Rupali Gund, Ravindra Zirmire, Haarshaadri J., Gaurav Kansagara and Colin Jamora, date: 01/20/2021, view: 19, Q&A: 0
    [Abstract]

    The skin is the largest organ that protects our body from the external environment and it is constantly exposed to pathogenic insults and injury. Repair of damage to this organ is carried out by a complex process involving three overlapping phases of inflammation, proliferation and remodeling. Histological analysis of wounded skin is a convenient

    ...
    Imaging of Human Cancer Cells in 3D Collagen Matrices
    Authors:  Karin Pfisterer, Brooke Lumicisi and Maddy Parsons, date: 01/20/2021, view: 18, Q&A: 0
    [Abstract]

    Research on cell migration and interactions with the extracellular matrix (ECM) was mostly focused on 2D surfaces in the past. Many recent studies have highlighted differences in migratory behaviour of cells on 2D surfaces compared to complex cell migration modes in 3D environments. When embedded in 3D matrices, cells constantly sense the

    ...
    Polyamine Transport Assay Using Reconstituted Yeast Membranes
    Authors:  Sarah van Veen, Shaun Martin, Marleen Schuermans and Peter Vangheluwe, date: 01/20/2021, view: 17, Q&A: 0
    [Abstract]

    ATP13A2/PARK9 is a late endo-/lysosomal P5B transport ATPase that is associated with several neurodegenerative disorders. We recently characterized ATP13A2 as a lysosomal polyamine exporter, which sheds light on the molecular identity of the unknown mammalian polyamine transport system. Here, we describe step by step a protocol to measure

    ...
    Determination of Antibacterial Activity of Film Coatings against Four Clinically Relevant Bacterial Strains
    [Abstract]

    Antibacterial coatings have currently gained great importance in biomedical technology investigations. Because of the spatial arrangement of the film coatings, evaluation of antibacterial activity presents a new challenge regarding traditional bacterial counting methods. In this protocol, four clinically relevant pathogens, Salmonella typhimurium,

    ...
    Wounding Caenorhabditis elegans with Glass Wool
    [Abstract]

    Research on wound healing majorly relies on rat, mice and other animal models. However, an alternative animal model ought to be brought in the field, pertaining to the stringent ethical issues owing to the use of animals in research. In this regard, Caenorhabdits elegans, a miniature model nematode gains the great attention of the researchers in

    ...
    Multiplex T-cell Stimulation Assay Utilizing a T-cell Activation Reporter-based Detection System
    Authors:  Laurie G. Landry, Sarah E. Mann, Amanda M. Anderson and Maki Nakayama, date: 01/20/2021, view: 29, Q&A: 0
    [Abstract]

    Immune tolerance and response are both largely driven by the interactions between the major histocompatibility complex (MHC) expressed by antigen presenting cells (APCs), T-cell receptors (TCRs) on T-cells, and their cognate antigens. Disordered interactions cause the pathogenesis of autoimmune diseases such as type 1 diabetes. Therefore, the

    ...
    A Parkinson’s Disease-relevant Mitochondrial and Neuronal Morphology High-throughput Screening Assay in LUHMES Cells
    [Abstract]

    Parkinson’s disease is a devastating neurodegenerative disorder affecting 2-3% of the population over 65 years of age. There is currently no disease-modifying treatment. One of the predominant pathological features of Parkinson’s disease is mitochondrial dysfunction, and much work has aimed to identify therapeutic compounds which can

    ...
    Stable Transformation of Arabidopsis thaliana Cell Suspension Cultures: A Case Study for the Overexpression of The COI1 Receptor
    [Abstract] Cell suspension cultures have been studied for decades to produce natural molecules. However, the difficulty in generating stably transformed cell lines has limited their use to produce high value chemicals reproducibly and in elevated quantities.

    In this protocol, a method to stably transform and maintain Arabidopsis cell ...



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