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      Effects of Non-Thermal Plasma on Mammalian Cells

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          Abstract

          Thermal plasmas and lasers have been widely used in medicine to cut, ablate and cauterize tissues through heating; in contrast, non-thermal plasma produces no heat, so its effects can be selective. In order to exploit the potential for clinical applications, including wound healing, sterilization, blood coagulation, and cancer treatment, a mechanistic understanding of the interaction of non-thermal plasma with living tissues is required. Using mammalian cells in culture, it is shown here that non-thermal plasma created by dielectric barrier discharge (DBD) has dose-dependent effects that range from increasing cell proliferation to inducing apoptosis. It is also shown that these effects are primarily due to formation of intracellular reactive oxygen species (ROS). We have utilized γ-H2AX to detect DNA damage induced by non-thermal plasma and found that it is initiated by production of active neutral species that most likely induce formation of organic peroxides in cell medium. Phosphorylation of H2AX following non-thermal plasma treatment is ATR dependent and ATM independent, suggesting that plasma treatment may lead to replication arrest or formation of single-stranded DNA breaks; however, plasma does not lead to formation of bulky adducts/thymine dimers.

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          Most cited references13

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          Apoptosis, oncosis, and necrosis. An overview of cell death.

          The historical development of the cell death concept is reviewed, with special attention to the origin of the terms necrosis, coagulation necrosis, autolysis, physiological cell death, programmed cell death, chromatolysis (the first name of apoptosis in 1914), karyorhexis, karyolysis, and cell suicide, of which there are three forms: by lysosomes, by free radicals, and by a genetic mechanism (apoptosis). Some of the typical features of apoptosis are discussed, such as budding (as opposed to blebbing and zeiosis) and the inflammatory response. For cell death not by apoptosis the most satisfactory term is accidental cell death. Necrosis is commonly used but it is not appropriate, because it does not indicate a form of cell death but refers to changes secondary to cell death by any mechanism, including apoptosis. Abundant data are available on one form of accidental cell death, namely ischemic cell death, which can be considered an entity of its own, caused by failure of the ionic pumps of the plasma membrane. Because ischemic cell death (in known models) is accompanied by swelling, the name oncosis is proposed for this condition. The term oncosis (derived from ónkos, meaning swelling) was proposed in 1910 by von Reckling-hausen precisely to mean cell death with swelling. Oncosis leads to necrosis with karyolysis and stands in contrast to apoptosis, which leads to necrosis with karyorhexis and cell shrinkage.
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            Histone H2AX is phosphorylated in an ATR-dependent manner in response to replicational stress.

            I Ward, J. Chen (2001)
            H2AX, a member of the histone H2A family, is rapidly phosphorylated in response to ionizing radiation. This phosphorylation, at an evolutionary conserved C-terminal phosphatidylinositol 3-OH-kinase-related kinase (PI3KK) motif, is thought to be critical for recognition and repair of DNA double strand breaks. Here we report that inhibition of DNA replication by hydroxyurea or ultraviolet irradiation also induces phosphorylation and foci formation of H2AX. These phospho-H2AX foci colocalize with proliferating cell nuclear antigen (PCNA), BRCA1, and 53BP1 at the arrested replication fork in S phase cells. This response is ATR-dependent but does not require ATM or Hus1. Our findings suggest that, in addition to its role in the recognition and repair of double strand breaks, H2AX also participates in the surveillance of DNA replication.
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              More than one way to die: apoptosis, necrosis and reactive oxygen damage.

              Cell death is an essential phenomenon in normal development and homeostasis, but also plays a crucial role in various pathologies. Our understanding of the molecular mechanisms involved has increased exponentially, although it is still far from complete. The morphological features of a cell dying either by apoptosis or by necrosis are remarkably conserved for quite different cell types derived from lower or higher organisms. At the molecular level, several gene products play a similar, crucial role in a major cell death pathway in a worm and in man. However, one should not oversimplify. It is now evident that there are multiple pathways leading to cell death, and some cells may have the required components for one pathway, but not for another, or contain endogenous inhibitors which preclude a particular pathway. Furthermore, different pathways can co-exist in the same cell and are switched on by specific stimuli. Apoptotic cell death, reported to be non-inflammatory, and necrotic cell death, which may be inflammatory, are two extremes, while the real situation is usually more complex. We here review the distinguishing features of the various cell death pathways: caspases (cysteine proteases cleaving after particular aspartate residues), mitochondria and/or reactive oxygen species are often, but not always, key components. As these various caspase-dependent and caspase-independent cell death pathways are becoming better characterized, we may learn to differentiate them, fill in the many gaps in our understanding, and perhaps exploit the knowledge acquired for clinical benefit.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2011
                21 January 2011
                : 6
                : 1
                : e16270
                Affiliations
                [1 ]Department of Electrical and Computer Engineering, Drexel University, Philadelphia, Pennsylvania, United States of America
                [2 ]Department of Biochemistry and Molecular Biology, Drexel University College of Medicine, Philadelphia, Pennsylvania, United States of America
                [3 ]Department of Surgery, Drexel University College of Medicine, Philadelphia, Pennsylvania, United States of America
                [4 ]Department of Mechanical Engineering and Mechanics, Drexel University, Philadelphia, Pennsylvania, United States of America
                Massachusetts Institute of Technology, United States of America
                Author notes

                Conceived and designed the experiments: SK CMK AF GF JAZ. Performed the experiments: SK CMK EC OA. Analyzed the data: SK CMK BZ OA AF GF JAZ. Wrote the paper: SK OA GF JAZ.

                Article
                PONE-D-10-03669
                10.1371/journal.pone.0016270
                3025030
                21283714
                36b564d3-235e-4aae-bab2-906782935c51
                Kalghatgi et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
                History
                : 10 October 2010
                : 9 December 2010
                Page count
                Pages: 11
                Categories
                Research Article
                Biology
                Biochemistry
                Nucleic Acids
                DNA
                DNA repair
                Biophysics
                Biomacromolecule-Ligand Interactions
                Protein Chemistry
                Biotechnology
                Bioengineering
                Biomedical Engineering
                Medical Devices
                Developmental Biology
                Molecular Development
                Signaling
                Molecular Cell Biology
                Gene Expression
                RNA interference
                Nucleic Acids
                DNA
                DNA repair
                Signal Transduction
                Signaling in Cellular Processes
                Signaling Pathways
                Cell Death
                Cell Growth
                Cellular Stress Responses
                Engineering
                Electrical Engineering
                Medicine
                Drugs and Devices
                Medical Devices
                Physics
                Plasmas

                Uncategorized
                Uncategorized

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