In vivo detection of free radicals in mouse septic encephalopathy using molecular MRI and immuno-spin trapping |
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| Affiliation: | 1. Advanced Magnetic Resonance Center, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA;2. Instituto de Pesquisa Clinica Evandro Chagas, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, RJ, Brazil;3. Laboratório de Bioquímica de Resposta ao Estresse, Programa de Biologia Molecular e Biotecnologia, Instituto de Bioquímica Médica, Universidade Federal do Rio de Janeiro, Cidade Universitária, Rio de Janeiro, RJ, Brazil;4. Laboratory of Pharmacology and Chemistry, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709, USA;5. Cardiovascular Biology, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA;6. Laboratory of Experimental Medicine & Therapeutics, Instituto Multidisciplinario de Investigaciones Biologicas–San Luis, CONICET, National University of San Luis, San Luis 5700, Argentina;7. Laboratório de Imunofarmacologia, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, RJ, Brazil;1. Pineal Research Laboratory, Department of Zoology, Banaras Hindu University, Varanasi 221 005, India;2. Departamento de Biología, Bioquímica y Farmacia, Universidad Nacional del Sur, 8000 Bahía Blanca, Argentina;3. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 11724;4. Department of Chemistry and Molecular and Cell Biology and the Howard Hughes Medical Institute, University of California, Berkeley, California 94720;5. Department of Surgery, Vermont Comprehensive Cancer Center, University of Vermont, College of Medicine, Burlington, Vermont 05405 |
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| Abstract: | Free radicals are known to play a major role in sepsis. Combined immuno-spin trapping and molecular magnetic resonance imaging (MRI) was used to detect in vivo and in situ levels of free radicals in murine septic encephalopathy after cecal ligation and puncture (CLP). DMPO (5,5-dimethyl pyrroline N-oxide) was injected over 6 h after CLP, before administration of an anti-DMPO probe (anti-DMPO antibody bound to albumin–gadolinium–diethylene triamine pentaacetic acid–biotin MRI targeting contrast agent). In vitro assessment of the anti-DMPO probe in oxidatively stressed mouse astrocytes significantly decreased T1 relaxation (p < 0.0001) compared to controls. MRI detected the presence of anti-DMPO adducts via a substantial decrease in %T1 change within the hippocampus, striatum, occipital, and medial cortex brain regions (p < 0.01 for all) in septic animals compared to shams, which was sustained for over 60 min (p < 0.05 for all). Fluorescently labeled streptavidin was used to target the anti-DMPO probe biotin, which was elevated in septic brain, liver, and lungs compared to sham. Ex vivo DMPO adducts (qualitative) and oxidative products, including 4-hydroxynonenal and 3-nitrotyrosine (quantitative, p < 0.05 for both), were elevated in septic brains compared to shams. This is the first study that has reported on the detection of in vivo and in situ levels of free radicals in murine septic encephalopathy. |
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| Keywords: | Immuno-spin trapping Free radicals Molecular MRI Sepsis Cecal ligation and puncture Mice in vivo DMPO Fluorescence microscopy 4-Hydroxynonenal 3-Nitrotyrosine |
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