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Reactive Oxygen Species: Molecules or ions formed by the incomplete one-electron reduction of oxygen. These reactive oxygen intermediates include Singlet oxygen; Superoxides; Peroxides; Hydroxyl radical; and Hypochlorous acid. They contribute to the microbicidal activity of Phagocytes, regulation of signal transduction and gene expression, and the oxidative damage to Nucleic acids; Proteins; and Lipids.

Detecting Reactive Oxygen Species

JoVE 5654

Reactive oxygen species are chemically active, oxygen-derived molecules capable of oxidizing other molecules. Because of their reactive nature, there are many deleterious effects associated with unchecked ROS production, including structural damage to DNA and other biological molecules. However, ROS can also be mediators of physiological signaling. There is accumulating evidence that ROS play significant roles in everything from activation of transcription factors to the mediation of inflammatory toxicity that kills foreign pathogens and defend the body.In this video we will delve into the associations between ROS, metabolism and disease. After establishing their significance, we will discuss the principles and a protocol of a commonly used methodology for measuring ROS levels in cells: the use of non-fluorescent probes that become fluorescent upon oxidation. Lastly, we will review some current applications of this technique in cell biology research.


 Cell Biology

An Introduction to Cell Metabolism

JoVE 5652

In cells, critical molecules are either built by joining together individual units like amino acids or nucleotides, or broken down into smaller components. Respectively, the reactions responsible for this are referred to as anabolic and catabolic. These reactions require or produce energy typically in the form of a “high-energy” molecule called ATP. Together, these processes make up “Cell Metabolism,” and are hallmarks of healthy, living cells.JoVE’s introduction to cell metabolism briefly reviews the rich history of this field, ranging from early studies on photosynthesis to more recent discoveries pertaining to energy production in all cells. This is followed by a discussion of some key questions asked by scientists studying metabolism, and common methods that they apply to answer these questions. Finally, we’ll explore how current researchers are studying alterations in metabolism that accompany metabolic disorders, or that occur following exposure to environmental stressors.


 Cell Biology

Introduction to the Microplate Reader

JoVE 5024

The microplate reader is a multimodal instrument that allows for a variety of experiments to be performed and measured simultaneously. Microplate readers can make absorbance, fluorescence and luminescence measurements. Multiwell plates are integral to the microplate reader and allow for many experiments to be performed at once. Regardless of the assay type, experiments on the plate reader utilize a standard curve to determine the experimental values. This curve uses samples of known concentration to generate a line of best fit or standard curve. Experimental values are then extrapolated to the curve or are calculated using the equation from the linear regression. Besides standards and samples being run on the multiwall plate, the blank along with positive and negative controls are also used in the assay to ensure it is working correctly. Multiplate readers are used to quantify protein, gene expression and various metabolic processes such as reactive oxygen species and calcium flux.


 General Laboratory Techniques

An Introduction to Aging and Regeneration

JoVE 5337

Tissues are maintained through a balance of cellular aging and regeneration. Aging refers to the gradual loss of cellular function, and regeneration is the repair of damaged tissue generally mediated by preexisting adult or somatic stem cells. Scientists are interested in understanding the biological mechanisms behind these two complex processes. By doing so, researchers may be able to use somatic stem cells to treat degenerative diseases and develop therapies that could delay the effects of aging. In this video, we provide a brief history of the field of aging and regeneration, touching upon observations made in ancient Greece, as well as modern-day experiments. Some of the questions being asked in this field, and the prominent methods being used by biologists to answer them, are then explored. Finally, we look at a few specific experiments being conducted in today\'s aging and regeneration research laboratories.


 Developmental Biology

Imaging Approaches to Assessments of Toxicological Oxidative Stress Using Genetically-encoded Fluorogenic Sensors

1Department of Environmental Sciences and Engineering, Gillings School of Global Public Health, University of North Carolina at Chapel Hill, 2Environmental Public Health Division, National Health and Environmental Effects Research Laboratory, U.S. Environmental Protection Agency, 3Oak Ridge Institute for Science and Education

JoVE 56945


 Cancer Research

Correlative Light Electron Microscopy (CLEM) for Tracking and Imaging Viral Protein Associated Structures in Cryo-immobilized Cells

1European Molecular Biology Laboratory, 2Department of Infectious Diseases, Molecular Virology, Heidelberg University, 3Central Facility for Electron Microscopy, Ulm University, 4Heidelberg Partner Site, German Center for Infection Research

JoVE 58154


 Bioengineering

Analyzing Beneficial Effects of Nutritional Supplements on Intestinal Epithelial Barrier Functions During Experimental Colitis

1Department of Molecular Biomedicine, Center for Research and Advanced Studies of the National Polytechnic Institute, 2Department of Physiology, Biophysics and Neurosciences, Center for Research and Advanced Studies of the National Polytechnic Institute, 3Department of Infectomics and Molecular Pathogenesis, Center for Research and Advanced Studies of the National Polytechnic Institute

JoVE 55095


 Medicine

Instrumentation of Near-term Fetal Sheep for Multivariate Chronic Non-anesthetized Recordings

1Département de sciences cliniques, CHUV, Université de Montréal, St-Hyacinthe, QC, 2Département d'obstetriques et de gynécologie, CHU Ste-Justine Research Centre, Université de Montréal, 3Département de neurosciences, CHU Ste-Justine Centre de recherche, Université de Montréal, 4Centre de recherche en reproduction animale (CRRA), Université de Montréal, St-Hyacinthe, QC

JoVE 52581


 Developmental Biology

Techniques for the Evolution of Robust Pentose-fermenting Yeast for Bioconversion of Lignocellulose to Ethanol

1Bioenergy Research Unit, National Center for Agricultural Utilization Research, 2Mycotoxin Prevention and Applied Microbiology Research Unit, National Center for Agricultural Utilization Research, 3Chemical Engineering and Material Science, Great Lakes Bioenergy Center, Michigan State University

JoVE 54227


 Bioengineering

In Vitro and In Vivo Detection of Mitophagy in Human Cells, C. Elegans, and Mice

1Laboratory of Molecular Gerontology, National Institute on Aging, National Institutes of Health, 2Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology - Hellas, 3Center for Molecular Medicine, National Heart Lung and Blood Institute, National Institutes of Health, 4Laboratory of Neurosciences, National Institute on Aging, National Institutes of Health, 5Nuffield Department of Obstetrics and Gynaecology, University of Oxford, 6Department of Clinical Molecular Biology, University of Oslo and Akershus University Hospital, 7Department of Basic Sciences, Faculty of Medicine, University of Crete, 8Danish Center for Healthy Aging, University of Copenhagen

JoVE 56301


 Medicine

Site-Directed Immobilization of Bone Morphogenetic Protein 2 to Solid Surfaces by Click Chemistry

1Fraunhofer-Institut für Grenzflächen- und Bioverfahrenstechnik (IGB), Translationszentrum Würzburg 'Regenerative Therapien für Krebs- und Muskuloskelettale Erkrankung', Institutsteil Würzburg, 2Lehrstuhl für Tissue Engineering und Regenerative Medizin, Universitätsklinikum Würzburg, 3Lehrstuhl für Pharmazeutische Technologie und Biopharmazie, Universität Würzburg, 4Lehrstuhl für molekulare Pflanzenphysiologie und Biophysik, Julius-von-Sachs Institut für Biowissenschaften, Universität Würzburg

JoVE 56616


 Bioengineering

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