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  1. AU="Nash, Kevin M"
  2. AU="Kubo, Sousuke"
  3. AU="Ingo Eitel"
  4. AU="van der Horst, A."
  5. AU="Di Mattia, A" AU="Di Mattia, A"
  6. AU="Di Pumpo, Marcello"
  7. AU="Doung, Yee-Cheen"
  8. AU="Saha, Moumita"
  9. AU="Wertz, Ashlee E"
  10. AU="Cowan, Michael J"
  11. AU=Togliatto Gabriele
  12. AU="Bassett, Dani S."
  13. AU="James Lemon"
  14. AU="Gros, Stephanie J"
  15. AU="Saeed Khademi"
  16. AU="Lallet-Daher, Helene"
  17. AU="Greenblatt, M"
  18. AU="Patwa, Ajay K"
  19. AU=Mastaglia F L
  20. AU="De Croock, Femke"
  21. AU=Robinson Michael J
  22. AU=Singh Romil
  23. AU="Martin, S J"
  24. AU="Szendrői, Miklós"
  25. AU="Moncel, Marie-Hélène"
  26. AU=Otu Akaninyene AU=Otu Akaninyene
  27. AU="Chiba, Kentaro"
  28. AU="Zhou, Jihua"
  29. AU="Ronald Bartels"
  30. AU="Liñares, J"
  31. AU="Valle, Valentina"
  32. AU="Tóth, András"
  33. AU="Pawar, Atul Darasing"
  34. AU="Semper, Chelsea"
  35. AU="Kraus, Joanne F"

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  1. Artikel: Current Perspectives on the Beneficial Role of Ginkgo biloba in Neurological and Cerebrovascular Disorders.

    Nash, Kevin M / Shah, Zahoor A

    Integrative medicine insights

    2015  Band 10, Seite(n) 1–9

    Abstract: Ginkgo biloba extract is an alternative medicine available as a standardized formulation, EGb 761(®), which consists of ginkgolides, bilobalide, and flavonoids. The individual constituents have varying therapeutic mechanisms that contribute to the ... ...

    Abstract Ginkgo biloba extract is an alternative medicine available as a standardized formulation, EGb 761(®), which consists of ginkgolides, bilobalide, and flavonoids. The individual constituents have varying therapeutic mechanisms that contribute to the pharmacological activity of the extract as a whole. Recent studies show anxiolytic properties of ginkgolide A, migraine with aura treatment by ginkgolide B, a reduction in ischemia-induced glutamate excitotoxicity by bilobalide, and an alternative antihypertensive property of quercetin, among others. These findings have been observed in EGb 761 as well and have led to clinical investigation into its use as a therapeutic for conditions such as cognition, dementia, cardiovascular, and cerebrovascular diseases. This review explores the therapeutic mechanisms of the individual EGb 761 constituents to explain the pharmacology as a whole and its clinical application to cardiovascular and neurological disorders, in particular ischemic stroke.
    Sprache Englisch
    Erscheinungsdatum 2015-11-09
    Erscheinungsland United States
    Dokumenttyp Journal Article ; Review
    ISSN 1177-3936
    ISSN 1177-3936
    DOI 10.4137/IMI.S25054
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  2. Artikel ; Online: Nanomedicine in the ROS-mediated pathophysiology: Applications and clinical advances.

    Nash, Kevin M / Ahmed, Salahuddin

    Nanomedicine : nanotechnology, biology, and medicine

    2015  Band 11, Heft 8, Seite(n) 2033–2040

    Abstract: Reactive oxygen species (ROS) are important in regulating normal cell physiological functions, but when produced in excess lead to the augmented pathogenesis of various diseases. Among these, ischemia reperfusion injury, Alzheimer's disease and ... ...

    Abstract Reactive oxygen species (ROS) are important in regulating normal cell physiological functions, but when produced in excess lead to the augmented pathogenesis of various diseases. Among these, ischemia reperfusion injury, Alzheimer's disease and rheumatoid arthritis are particularly important. Since ROS can be counteracted by a variety of antioxidants, natural and synthetic antioxidants have been developed. However, due to the ubiquitous production of ROS in living systems, poor in vivo efficiency of these agents and lack of target specificity, the current clinical modalities to treat oxidative stress damage are limited. Advances in the developing field of nanomedicine have yielded nanoparticles that can prolong antioxidant activity, and target specificity of these agents. This article reviews recent advances in antioxidant nanoparticles and their applications to manage oxidative stress-mediated diseases.
    From the clinical editor: Production of reactive oxygen species (ROS) is a purely physiological process in many disease conditions. However, excessive and uncontrolled production will lead to oxidative stress and further tissue damage. Advances in nanomedicine have provided many novel strategies to try to combat and counteract ROS. In this review article, the authors comprehensively highlighted the current status and future developments in using nanotechnology for providing novel therapeutic options in this field.
    Mesh-Begriff(e) Animals ; Antioxidants/administration & dosage ; Antioxidants/chemistry ; Antioxidants/therapeutic use ; Drug Carriers/chemistry ; Drug Delivery Systems/methods ; Humans ; Nanomedicine/methods ; Nanoparticles/chemistry ; Nanotechnology/methods ; Oxidative Stress/drug effects ; Reactive Oxygen Species/metabolism
    Chemische Substanzen Antioxidants ; Drug Carriers ; Reactive Oxygen Species
    Sprache Englisch
    Erscheinungsdatum 2015-08-06
    Erscheinungsland United States
    Dokumenttyp Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Review
    ZDB-ID 2183417-9
    ISSN 1549-9642 ; 1549-9634
    ISSN (online) 1549-9642
    ISSN 1549-9634
    DOI 10.1016/j.nano.2015.07.003
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  3. Artikel ; Online: Evaluation of tissue-engineered human acellular vessels as a Blalock-Taussig-Thomas shunt in a juvenile primate model.

    Nash, Kevin M / Boe, Brian A / Carrillo, Sergio A / Harrison, Andrew / Iwaki, Ryuma / Kelly, John / Kirkton, Robert D / Krishnamurthy, Ramkumar / Lawson, Jeffrey H / Matsuzaki, Yuichi / Prichard, Heather L / Shah, Kejal / Shinoka, Toshiharu / Breuer, Christopher K

    JTCVS open

    2023  Band 15, Seite(n) 433–445

    Abstract: Objectives: Palliative treatment of cyanotic congenital heart disease (CCHD) uses systemic-to-pulmonary conduits, often a modified Blalock-Taussig-Thomas shunt (mBTTs). Expanded polytetrafluoroethylene (ePTFE) mBTTs have associated risks for thrombosis ... ...

    Abstract Objectives: Palliative treatment of cyanotic congenital heart disease (CCHD) uses systemic-to-pulmonary conduits, often a modified Blalock-Taussig-Thomas shunt (mBTTs). Expanded polytetrafluoroethylene (ePTFE) mBTTs have associated risks for thrombosis and infection. The Human Acellular Vessel (HAV) (Humacyte, Inc) is a decellularized tissue-engineered blood vessel currently in clinical trials in adults for vascular trauma, peripheral artery disease, and end-stage renal disease requiring hemodialysis. In addition to restoring blood flow, the engineered HAV demonstrates the capacity for host cellular remodeling into native-like vasculature. Here we report preclinical evaluation of a small-diameter (3.5 mm) HAV as a mBTTs in a non-human primate model.
    Methods: We implanted 3.5 mm HAVs as right subclavian artery to pulmonary artery mBTTs in non-immunosuppressed juvenile rhesus macaques (n = 5). HAV patency, structure, and blood flow were assessed by postoperative imaging from 1 week to 6 months. Histology of HAVs and surrounding tissues was performed.
    Results: Surgical procedures were well tolerated, with satisfactory anastomoses, showing feasibility of using the 3.5 mm HAV as a mBTTs. All macaques had some immunological reactivity to the human extracellular matrix, as expected in this xenogeneic model. HAV mBTTs remained patent for up to 6 months in animals, exhibiting mild immunoreactivity. Two macaques displaying more severe immunoreactivity to the human HAV material developed midgraft dilatation without bleeding or rupture. HAV repopulation by host cells expressing smooth muscle and endothelial markers was observed in all animals.
    Conclusions: These findings may support use of 3.5 mm HAVs as mBTTs in CCHD and potentially other pediatric vascular indications.
    Sprache Englisch
    Erscheinungsdatum 2023-08-09
    Erscheinungsland Netherlands
    Dokumenttyp Journal Article
    ISSN 2666-2736
    ISSN (online) 2666-2736
    DOI 10.1016/j.xjon.2023.05.018
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  4. Artikel ; Online: Development of a reactive oxygen species-sensitive nitric oxide synthase inhibitor for the treatment of ischemic stroke.

    Nash, Kevin M / Schiefer, Isaac T / Shah, Zahoor A

    Free radical biology & medicine

    2017  Band 115, Seite(n) 395–404

    Abstract: Ischemic stroke is caused by a blockage of cerebral blood flow resulting in neuronal and glial hypoxia leading to inflammatory and reactive oxygen species (ROS)-mediated cell death. Nitric oxide (NO) formed by NO synthase (NOS) is known to be protective ... ...

    Abstract Ischemic stroke is caused by a blockage of cerebral blood flow resulting in neuronal and glial hypoxia leading to inflammatory and reactive oxygen species (ROS)-mediated cell death. Nitric oxide (NO) formed by NO synthase (NOS) is known to be protective in ischemic stroke, however NOS has been shown to 'uncouple' under oxidative conditions to instead produce ROS. Nitrones are antioxidant molecules that are shown to trap ROS to then decompose and release NO. In this study, the nitrone 5 was designed such that its decomposition product is a NOS inhibitor, 6, effectively leading to NOS inhibition specifically at the site of ROS production. The ability of 5 to spin-trap radicals and decompose to 6 was observed using EPR and LC-MS/MS. The pro-drug concept was tested in vitro by measuring cell viability and 6 formation in SH-SY5Y cells subjected to oxygen glucose deprivation (OGD). 5 was found to be more efficacious and more potent than PBN, and was able to increase phospho-Akt while reducing nitrotyrosine and cleaved caspase-3 levels. 6 treatment, but not 5, was found to decrease NO production in LPS-stimulated microglia. Doppler flowmetry on anesthetized mice showed increased cerebral blood flow upon intravenous administration of 1mg/kg of 5, but a return to baseline upon administration of 10mg/kg, likely due to its dual nature of antioxidant/NO-donor and NOS-inhibition. Mice treated with 5 after permanent ischemia exhibited a >30% reduction in infarct volume, and higher formation of 6 in ischemic tissue resulting in region specific effects limited to the infarct area.
    Mesh-Begriff(e) Animals ; Antioxidants/chemical synthesis ; Antioxidants/therapeutic use ; Caspase 3/metabolism ; Cell Survival ; Cells, Cultured ; Disease Models, Animal ; Humans ; Ischemia/drug therapy ; Mice ; Mice, Inbred C57BL ; Microglia/drug effects ; Microglia/physiology ; Neurons/drug effects ; Neurons/physiology ; Nitric Oxide/metabolism ; Nitric Oxide Synthase/antagonists & inhibitors ; Reactive Oxygen Species/metabolism ; Stroke/drug therapy ; Tyrosine/analogs & derivatives ; Tyrosine/metabolism
    Chemische Substanzen Antioxidants ; Reactive Oxygen Species ; Nitric Oxide (31C4KY9ESH) ; 3-nitrotyrosine (3604-79-3) ; Tyrosine (42HK56048U) ; Nitric Oxide Synthase (EC 1.14.13.39) ; Caspase 3 (EC 3.4.22.-)
    Sprache Englisch
    Erscheinungsdatum 2017-12-22
    Erscheinungsland United States
    Dokumenttyp Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 807032-5
    ISSN 1873-4596 ; 0891-5849
    ISSN (online) 1873-4596
    ISSN 0891-5849
    DOI 10.1016/j.freeradbiomed.2017.12.027
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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    Verfügbar in ZB MED Köln/Königswinter

    Zs.A 2109: Hefte anzeigen Standort:
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    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
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  5. Artikel ; Online: Type-I diabetes aggravates post-hemorrhagic stroke cognitive impairment by augmenting oxidative stress and neuroinflammation in mice.

    Bahader, Ghaith A / Nash, Kevin M / Almarghalani, Daniyah A / Alhadidi, Qasim / McInerney, Marcia F / Shah, Zahoor A

    Neurochemistry international

    2021  Band 149, Seite(n) 105151

    Abstract: Diabetes Mellitus (DM) is a major comorbid condition that increases susceptibility to stroke. Intracerebral hemorrhage (ICH), a devastating type of stroke, accounts for only 13% of the total stroke cases but is associated with higher mortality. ... ...

    Abstract Diabetes Mellitus (DM) is a major comorbid condition that increases susceptibility to stroke. Intracerebral hemorrhage (ICH), a devastating type of stroke, accounts for only 13% of the total stroke cases but is associated with higher mortality. Multimorbid models of DM and ischemic stroke have been widely studied; however, fewer pieces of evidence are available on the impact of DM on the outcomes of ICH injury. In this study, we investigated the effect of DM on ICH-induced injury and cognitive impairments. Streptozotocin (STZ) induced type-I DM (T1DM) animal model was used, and experimental ICH was induced by intrastriatal injection of collagenase. Our results demonstrated that DM is associated with a significant increase in hematoma volume and deficits in post-stroke locomotor, sensorimotor, and cognitive behavior in mice. The levels of neuroinflammation, oxidative/nitrosative stress, and glial cell activation were also increased in the diabetic mice following ICH injury. This study provides a better understanding of the influence of DM comorbidity on hemorrhagic stroke outcomes and uncovers the important pathological mechanisms underlying DM-induced exacerbation of ICH injury.
    Mesh-Begriff(e) Animals ; Cerebral Hemorrhage/chemically induced ; Cerebral Hemorrhage/metabolism ; Cognitive Dysfunction/chemically induced ; Cognitive Dysfunction/metabolism ; Diabetes Mellitus, Experimental/chemically induced ; Diabetes Mellitus, Experimental/metabolism ; Diabetes Mellitus, Type 1/chemically induced ; Diabetes Mellitus, Type 1/metabolism ; Hand Strength/physiology ; Inflammation Mediators/metabolism ; Locomotion/drug effects ; Locomotion/physiology ; Male ; Maze Learning/physiology ; Mice ; Mice, Inbred C57BL ; Oxidative Stress/physiology ; Streptozocin/toxicity ; Stroke/chemically induced ; Stroke/metabolism
    Chemische Substanzen Inflammation Mediators ; Streptozocin (5W494URQ81)
    Sprache Englisch
    Erscheinungsdatum 2021-08-01
    Erscheinungsland England
    Dokumenttyp Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 283190-9
    ISSN 1872-9754 ; 0197-0186
    ISSN (online) 1872-9754
    ISSN 0197-0186
    DOI 10.1016/j.neuint.2021.105151
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    Verfügbar in ZB MED Köln/Königswinter

    Zs.A 1610: Hefte anzeigen Standort:
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  6. Artikel ; Online: Cofilin Knockdown Attenuates Hemorrhagic Brain Injury-induced Oxidative Stress and Microglial Activation in Mice.

    Alhadidi, Qasim / Nash, Kevin M / Alaqel, Saleh / Sayeed, Muhammad Shahdaat Bin / Shah, Zahoor A

    Neuroscience

    2018  Band 383, Seite(n) 33–45

    Abstract: Intracerebral hemorrhage (ICH) resulting from the rupture of the blood vessels in the brain is associated with significantly higher mortality and morbidity. Clinical studies focused on alleviating the primary injury, hematoma formation and expansion, ... ...

    Abstract Intracerebral hemorrhage (ICH) resulting from the rupture of the blood vessels in the brain is associated with significantly higher mortality and morbidity. Clinical studies focused on alleviating the primary injury, hematoma formation and expansion, were largely ineffective, suggesting that secondary injury-induced inflammation and the formation of reactive species also contribute to the overall injury process. In this study, we explored the effects of cofilin knockdown in a mouse model of ICH. Animals given stereotaxic injections of cofilin siRNA, 72-h prior to induction of ICH by collagenase injection within the area of siRNA administration showed significantly decreased cofilin expression levels and lower hemorrhage volume and edema, and the animals performed significantly better in neurobehavioral tasks i.e., rotarod, grip strength and neurologic deficit scores. Cofilin siRNA knocked-down mice had reduced ICH-induced DNA fragmentation, blood-brain barrier disruption and microglial activation, with a concomitant increase in astrocyte activation. Increased expression of pro-survival proteins and decreased markers of oxidative stress were also observed in cofilin siRNA-treated mice possibly due to the reduced levels of cofilin. Our results suggest that cofilin plays a major role in ICH-induced secondary injury, and could become a potential therapeutic target.
    Mesh-Begriff(e) Actin Depolymerizing Factors/metabolism ; Animals ; Gene Knockdown Techniques ; Intracranial Hemorrhages/metabolism ; Intracranial Hemorrhages/pathology ; Intracranial Hemorrhages/physiopathology ; Male ; Mice ; Mice, Inbred C57BL ; Microglia/pathology ; Oxidative Stress/physiology
    Chemische Substanzen Actin Depolymerizing Factors
    Sprache Englisch
    Erscheinungsdatum 2018-05-08
    Erscheinungsland United States
    Dokumenttyp Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 196739-3
    ISSN 1873-7544 ; 0306-4522
    ISSN (online) 1873-7544
    ISSN 0306-4522
    DOI 10.1016/j.neuroscience.2018.04.036
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    Zs.A 1215: Hefte anzeigen Standort:
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  7. Artikel ; Online: Potential implication of the chemical properties and bioactivity of nitrone spin traps for therapeutics.

    Villamena, Frederick A / Das, Amlan / Nash, Kevin M

    Future medicinal chemistry

    2012  Band 4, Heft 9, Seite(n) 1171–1207

    Abstract: Nitrone therapeutics has been employed in the treatment of oxidative stress-related diseases such as neurodegeneration, cardiovascular disease and cancer. The nitrone-based compound NXY-059, which is the first drug to reach clinical trials for the ... ...

    Abstract Nitrone therapeutics has been employed in the treatment of oxidative stress-related diseases such as neurodegeneration, cardiovascular disease and cancer. The nitrone-based compound NXY-059, which is the first drug to reach clinical trials for the treatment of acute ischemic stroke, has provided promise for the development of more robust pharmacological agents. However, the specific mechanism of nitrone bioactivity remains unclear. In this review, we present a variety of nitrone chemistry and biological activity that could be implicated for the nitrone's pharmacological activity. The chemistries of spin trapping and spin adduct reveal insights on the possible roles of nitrones for altering cellular redox status through radical scavenging or nitric oxide donation, and their biological effects are presented. An interdisciplinary approach towards the development of novel synthetic antioxidants with improved pharmacological properties encompassing theoretical, synthetic, biochemical and in vitro/in vivo studies is covered.
    Mesh-Begriff(e) Benzenesulfonates/chemistry ; Benzenesulfonates/metabolism ; Free Radical Scavengers/metabolism ; Spin Labels ; Superoxides/metabolism
    Chemische Substanzen Benzenesulfonates ; Free Radical Scavengers ; Spin Labels ; Superoxides (11062-77-4) ; disufenton sodium (7M1J3HN9VO)
    Sprache Englisch
    Erscheinungsdatum 2012-06-18
    Erscheinungsland England
    Dokumenttyp Journal Article ; Research Support, N.I.H., Extramural ; Review
    ISSN 1756-8927
    ISSN (online) 1756-8927
    DOI 10.4155/fmc.12.74
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  8. Artikel ; Online: Reactive nitrogen species reactivities with nitrones: theoretical and experimental studies.

    Nash, Kevin M / Rockenbauer, Antal / Villamena, Frederick A

    Chemical research in toxicology

    2012  Band 25, Heft 8, Seite(n) 1581–1597

    Abstract: Reactive nitrogen species (RNS) such as nitrogen dioxide ((•)NO(2)), peroxynitrite (ONOO(-)), and nitrosoperoxycarbonate (ONOOCO(2)(-)) are among the most damaging species present in biological systems due to their ability to cause modification of key ... ...

    Abstract Reactive nitrogen species (RNS) such as nitrogen dioxide ((•)NO(2)), peroxynitrite (ONOO(-)), and nitrosoperoxycarbonate (ONOOCO(2)(-)) are among the most damaging species present in biological systems due to their ability to cause modification of key biomolecular systems through oxidation, nitrosylation, and nitration. Nitrone spin traps are known to react with free radicals and nonradicals via electrophilic and nucleophilic addition reactions and have been employed as reagents to detect radicals using electron paramagnetic resonance (EPR) spectroscopy and as pharmacological agents against oxidative stress-mediated injury. This study examines the reactivity of cyclic nitrones such as 5,5-dimethylpyrroline N-oxide (DMPO) with (•)NO(2), ONOO(-), ONOOCO(2)(-), SNAP, and SIN-1 using EPR. The thermochemistries of nitrone reactivity with RNS and isotropic hfsc's of the addition products were also calculated at the PCM(water)/B3LYP/6-31+G**//B3LYP/6-31G* level of theory with and without explicit water molecules to rationalize the nature of the observed EPR spectra. Spin trapping of other RNS such as azide ((•)N(3)), nitrogen trioxide ((•)NO(3)), amino ((•)NH(2)) radicals and nitroxyl (HNO) were also theoretically and experimentally investigated by EPR spin trapping and mass spectrometry. This study also shows that other spin traps such as 5-carbamoyl-5-methyl-pyrroline N-oxide, 5-ethoxycarbonyl-5-methyl-pyrroline N-oxide, and 5-(diethoxyphosphoryl)-5-methyl-1-pyrroline N-oxide can react with radical and nonradical RNS, thus making spin traps suitable probes as well as antioxidants against RNS-mediated oxidative damage.
    Mesh-Begriff(e) Carbonates/chemistry ; Cyclic N-Oxides/chemistry ; Electron Spin Resonance Spectroscopy ; Mass Spectrometry ; Models, Molecular ; Nitrates/chemistry ; Nitrogen Oxides/chemistry ; Oxidation-Reduction ; Peroxynitrous Acid/chemistry ; Reactive Nitrogen Species/chemistry ; Spin Trapping ; Thermodynamics
    Chemische Substanzen Carbonates ; Cyclic N-Oxides ; Nitrates ; Nitrogen Oxides ; Reactive Nitrogen Species ; nitrones ; nitrosoperoxycarbonate ; Peroxynitrous Acid (14691-52-2)
    Sprache Englisch
    Erscheinungsdatum 2012-07-31
    Erscheinungsland United States
    Dokumenttyp Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 639353-6
    ISSN 1520-5010 ; 0893-228X
    ISSN (online) 1520-5010
    ISSN 0893-228X
    DOI 10.1021/tx200526y
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  9. Artikel ; Online: Detection of nitric oxide and superoxide radical anion by electron paramagnetic resonance spectroscopy from cells using spin traps.

    Gopalakrishnan, Bhavani / Nash, Kevin M / Velayutham, Murugesan / Villamena, Frederick A

    Journal of visualized experiments : JoVE

    2012  , Heft 66, Seite(n) e2810

    Abstract: Reactive nitrogen/oxygen species (ROS/RNS) at low concentrations play an important role in regulating cell function, signaling, and immune response but in unregulated concentrations are detrimental to cell viability. While living systems have evolved ... ...

    Abstract Reactive nitrogen/oxygen species (ROS/RNS) at low concentrations play an important role in regulating cell function, signaling, and immune response but in unregulated concentrations are detrimental to cell viability. While living systems have evolved with endogenous and dietary antioxidant defense mechanisms to regulate ROS generation, ROS are produced continuously as natural by-products of normal metabolism of oxygen and can cause oxidative damage to biomolecules resulting in loss of protein function, DNA cleavage, or lipid peroxidation, and ultimately to oxidative stress leading to cell injury or death. Superoxide radical anion (O2•-) is the major precursor of some of the most highly oxidizing species known to exist in biological systems such as peroxynitrite and hydroxyl radical. The generation of O2•- signals the first sign of oxidative burst, and therefore, its detection and/or sequestration in biological systems is important. In this demonstration, O2•- was generated from polymorphonuclear neutrophils (PMNs). Through chemotactic stimulation with phorbol-12-myristate-13-acetate (PMA), PMN generates O2•- via activation of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase. Nitric oxide (NO) synthase which comes in three isoforms, as inducible-, neuronal- and endothelial-NOS, or iNOS, nNOS or eNOS, respectively, catalyzes the conversion of L- arginine to L-citrulline, using NADPH to produce NO. Here, we generated NO from endothelial cells. Under oxidative stress conditions, eNOS for example can switch from producing NO to O2•- in a process called uncoupling, which is believed to be caused by oxidation of heme or the co-factor, tetrahydrobiopterin (BH4). There are only few reliable methods for the detection of free radicals in biological systems but are limited by specificity and sensitivity. Spin trapping is commonly used for the identification of free radicals and involves the addition reaction of a radical to a spin trap forming a persistent spin adduct which can be detected by electron paramagnetic resonance (EPR) spectroscopy. The various radical adducts exhibit distinctive spectrum which can be used to identify the radicals being generated and can provide a wealth of information about the nature and kinetics of radical production. The cyclic nitrones, 5,5-dimethyl-pyrroline-N-oxide, DMPO, the phosphoryl-substituted DEPMPO, and the ester-substituted, EMPO and BMPO, have been widely employed as spin traps--the latter spin traps exhibiting longer half-lives for O2•- adduct. Iron (II)-N-methyl-D-glucamine dithiocarbamate, Fe(MGD)2 is commonly used to trap NO due to high rate of adduct formation and the high stability of the spin adduct.
    Mesh-Begriff(e) Animals ; Cattle ; Electron Spin Resonance Spectroscopy/methods ; Endothelial Cells/chemistry ; Nitric Oxide/analysis ; Nitric Oxide/chemistry ; Nitrogen Oxides/chemistry ; Spin Trapping/methods ; Superoxides/analysis ; Superoxides/chemistry
    Chemische Substanzen Nitrogen Oxides ; nitrones ; Superoxides (11062-77-4) ; Nitric Oxide (31C4KY9ESH)
    Sprache Englisch
    Erscheinungsdatum 2012-08-18
    Erscheinungsland United States
    Dokumenttyp Journal Article ; Research Support, N.I.H., Extramural ; Video-Audio Media
    ZDB-ID 2259946-0
    ISSN 1940-087X ; 1940-087X
    ISSN (online) 1940-087X
    ISSN 1940-087X
    DOI 10.3791/2810
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  10. Artikel: Detection of nitric oxide and superoxide radical anion by electron paramagnetic resonance spectroscopy from cells using spin traps

    Gopalakrishnan, Bhavani / Nash, Kevin M / Velayutham, Murugesan / Villamena, Frederick A

    Journal of visualized experiments. 2012 Aug. 18, , no. 66

    2012  

    Abstract: Reactive nitrogen/oxygen species (ROS/RNS) at low concentrations play an important role in regulating cell function, signaling, and immune response but in unregulated concentrations are detrimental to cell viability1, 2. While living systems have evolved ...

    Abstract Reactive nitrogen/oxygen species (ROS/RNS) at low concentrations play an important role in regulating cell function, signaling, and immune response but in unregulated concentrations are detrimental to cell viability1, 2. While living systems have evolved with endogenous and dietary antioxidant defense mechanisms to regulate ROS generation, ROS are produced continuously as natural by-products of normal metabolism of oxygen and can cause oxidative damage to biomolecules resulting in loss of protein function, DNA cleavage, or lipid peroxidation3, and ultimately to oxidative stress leading to cell injury or death4. Superoxide radical anion (O2•-) is the major precursor of some of the most highly oxidizing species known to exist in biological systems such as peroxynitrite and hydroxyl radical. The generation of O2•- signals the first sign of oxidative burst, and therefore, its detection and/or sequestration in biological systems is important. In this demonstration, O2•- was generated from polymorphonuclear neutrophils (PMNs). Through chemotactic stimulation with phorbol-12-myristate-13-acetate (PMA), PMN generates O2•- via activation of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase5. Nitric oxide (NO) synthase which comes in three isoforms, as inducible-, neuronal- and endothelial-NOS, or iNOS, nNOS or eNOS, respectively, catalyzes the conversion of L- arginine to L-citrulline, using NADPH to produce NO6. Here, we generated NO from endothelial cells. Under oxidative stress conditions, eNOS for example can switch from producing NO to O2•- in a process called uncoupling, which is believed to be caused by oxidation of heme7 or the co-factor, tetrahydrobiopterin (BH4)8. There are only few reliable methods for the detection of free radicals in biological systems but are limited by specificity and sensitivity. Spin trapping is commonly used for the identification of free radicals and involves the addition reaction of a radical to a spin trap forming a persistent spin adduct which can be detected by electron paramagnetic resonance (EPR) spectroscopy. The various radical adducts exhibit distinctive spectrum which can be used to identify the radicals being generated and can provide a wealth of information about the nature and kinetics of radical production9. The cyclic nitrones, 5,5-dimethyl-pyrroline-N-oxide, DMPO10, the phosphoryl-substituted DEPMPO11, and the ester-substituted, EMPO12 and BMPO13, have been widely employed as spin traps--the latter spin traps exhibiting longer half-lives for O2•- adduct. Iron (II)-N-methyl-D-glucamine dithiocarbamate, Fe(MGD)2 is commonly used to trap NO due to high rate of adduct formation and the high stability of the spin adduct14.
    Schlagwörter DNA damage ; NADP (coenzyme) ; antioxidant activity ; arginine ; byproducts ; catalytic activity ; chemotaxis ; citrulline ; defense mechanisms ; electron paramagnetic resonance spectroscopy ; endothelial cells ; endothelial nitric oxide synthase ; free radicals ; half life ; hydroxyl radicals ; immune response ; inducible nitric oxide synthase ; iron ; lipids ; metabolism ; neuronal nitric oxide synthase ; neurons ; neutrophils ; nitric oxide ; nitrogen ; oxidation ; oxidative stress ; oxygen ; spin trapping ; superoxide anion
    Sprache Englisch
    Erscheinungsverlauf 2012-0818
    Umfang p. e2810.
    Erscheinungsort Journal of Visualized Experiments
    Dokumenttyp Artikel
    ZDB-ID 2259946-0
    ISSN 1940-087X
    ISSN 1940-087X
    DOI 10.3791/2810
    Datenquelle NAL Katalog (AGRICOLA)

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