Protocols in Current Issue
    Live Mitochondrial or Cytosolic Calcium Imaging Using Genetically-encoded Cameleon Indicator in Mammalian Cells
    Authors:  Elisa Greotti and Tullio Pozzan, date: 02/05/2020, view: 291, Q&A: 0
    [Abstract] Calcium (Ca2+) imaging aims at investigating the dynamic changes in live cells of its concentration ([Ca2+]) in different pathophysiological conditions. Ca2+ is an ubiquitous and versatile intracellular signal that modulates a large variety of cellular functions thanks to a cell type-specific toolkit and a complex ...
    Measurement of Chloroplastic NAD Kinase Activity and Whole Tissue NAD Kinase Assay
    Authors:  Yuuma Ishikawa, Maki Kawai-Yamada and Shin-nosuke Hashida, date: 01/05/2020, view: 394, Q&A: 0
    [Abstract] Nicotinamide adenine dinucleotide phosphate (NADP) synthesis requires nicotinamide adenine dinucleotide (NAD) kinase activity, substrate NAD and ATP. The NAD kinase responds to various environmental stimuli and its activity is regulated via various regulatory pathways, such as Ca2+-dependent and redox-dependent signals. Conventional in ...
    Intracellular cAMP Measurements in Candida albicans Biofilms
    Authors:  Liuliu Jiang, Shengyan Chen, Kairui Sun, Peng Zhou and Xin Wei, date: 12/05/2019, view: 436, Q&A: 0
    [Abstract] Candida albicans is the most common cause of fungal infections worldwide. Infection by C. albicans is closely associated with its ability to form a biofilm, closely packed communities of cells attached to the surfaces of human tissues and implanted devices, in or on the host. When tested for susceptibility to antifungals, such as ...
    Amplex Red Assay for Measuring Hydrogen Peroxide Production from Caenorhabditis elegans
    Authors:  Ozgur Karakuzu, Melissa R. Cruz, Yi Liu and Danielle A. Garsin, date: 11/05/2019, view: 895, Q&A: 0
    [Abstract] Reagents such as Amplex® Red have been developed for detecting hydrogen peroxide (H2O2) and are used to measure the release of H2O2 from biological samples such as mammalian leukocytes undergoing the oxidative burst. Caenorhabditis elegans is commonly used as a model host in the study ...
    Isolation, Purification, and Characterization of Ginger-derived Nanoparticles (GDNPs) from Ginger, Rhizome of Zingiber officinale
    Authors:  Junsik Sung, Chunhua Yang, Emilie Viennois, Mingzhen Zhang and Didier Merlin, date: 10/05/2019, view: 986, Q&A: 1
    [Abstract] Factors implicated in the pathophysiology of intestinal inflammation include defects in intestinal epithelial barrier function, abnormal immune responses, and activities of the gut microbiota. Current agents used to treat human Inflammatory Bowels Disease (IBD), chronic inflammation of digestive tract, have serious side effects. In addition, most ...
    Photoaffinity Labeling of Respiratory Complex I in Bovine Heart Submitochondrial Particles by Photoreactive [125I] amilorides
    Authors:  Masatoshi Murai and Hideto Miyoshi, date: 09/05/2019, view: 627, Q&A: 0
    [Abstract] The architecture of quinone/inhibitor-access channel in proton-translocating NADH-quinone oxidoreductase (respiratory complex I) was modeled by X-ray crystallography and cryo-EM, however, it remains debatable whether the channel model reflects the physiologically relevant state present throughout the catalytic cycle. Using photoreactive [125 ...
    Quantification of Prostaglandin E2 Concentration in Interstitial Fluid from the Hypothalamic Region of Free-moving Mice
    Authors:  Yoshinobu Nakamura, Takatoshi Sakaguchi, Ikumi Tamai and Takeo Nakanishi, date: 08/05/2019, view: 753, Q&A: 0
    [Abstract] Prostaglandin E2 (PGE2) is a well-established chemical mediator for the generation of the fever at the hypothalamus of the brain. PGE2 mediates fever generation via PGE receptor 3 (i.e., EP3) on neurons in the preoptic area. The role of PGE2 has been analyzed by measuring PGE2 ...
    Oxygen Consumption Measurements in Caenorhabditis elegans Using the Seahorse XF24
    Authors:  Suraiya Haroon and Marc Vermulst, date: 07/05/2019, view: 1186, Q&A: 0
    [Abstract] Mitochondria generate 90% of the energy required to sustain life. As a result, loss of mitochondrial function compromises almost every facet of human physiology. Accordingly, most mitochondrial diseases tend to present themselves as complex, multi-systemic disorders that can be difficult to diagnose. Depending on the severity of the mitochondrial ...
    Simple Method to Determine Protein Redox State in Arabidopsis thaliana
    Authors:  Keisuke Yoshida and Toru Hisabori, date: 06/05/2019, view: 1895, Q&A: 0
    [Abstract] Thiol-based redox regulation is a posttranslational protein modification that plays a key role in many biological aspects. To understand its regulatory functions, we need a method to directly assess protein redox state in vivo. Here we present a simple procedure to determine protein redox state in a model plant Arabidopsis thaliana ...
    Hypochlorous Acid Staining with R19-S in the Drosophila Intestine upon Ingestion of Opportunistic Bacteria
    Authors:  Salma Hachfi, Olivia Benguettat and Armel Gallet, date: 05/20/2019, view: 1521, Q&A: 0
    [Abstract] The intestine is endowed with an innate immune system that is required to fight any exogenous bacteria that are swallowed along with the food. The first line of defense that is mounted by the gut epithelium is the release of immune Reactive Oxygen Species (ROS), such as hypochlorous acid (HOCl), into the lumen. HOCl is produced within 1.5 h of ...

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