RESUMEN
BACKGROUND: Inhalation of thermal water with antioxidant properties is empirically used for COPD. AIMS: To evaluate the effects of sulphurous thermal water (reducing agents) on airway oxidant stress and clinical outcomes in COPD. METHODS: Forty moderate-to-severe COPD patients were randomly assigned to receive 12-day inhalation with sulphurous thermal water or isotonic saline. Patients were assessed for superoxide anion (O2 (-)) production in the exhaled breath condensate and clinical outcomes at recruitment, the day after the conclusion of the 12-day inhalation treatment, and one month after the end of the inhalation treatment. RESULTS: Inhalation of reducing agents resulted in a significant reduction of O2 (-) production in exhaled breath condensate of COPD patients at the end of the inhalatory treatment and at followup compared to baseline. A significant improvement in the COPD assessment test (CAT) questionnaire was shown one month after the end of the inhalatory treatment only in patients receiving sulphurous water. CONCLUSION: Thermal water inhalation produced an in vivo antioxidant effect and improvement in health status in COPD patients. Larger studies are required in order to evaluate whether inhalation of thermal water is able to modify relevant clinical outcomes of the disease (the study was registered at clinicaltrial.gov-identifier: NCT01664767).
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Enfermedad Pulmonar Obstructiva Crónica/tratamiento farmacológico , Sustancias Reductoras/uso terapéutico , Estallido Respiratorio/efectos de los fármacos , Azufre/administración & dosificación , Administración por Inhalación , Anciano , Anciano de 80 o más Años , Antioxidantes/uso terapéutico , Pruebas Respiratorias , Femenino , Humanos , Masculino , Persona de Mediana Edad , Enfermedad Pulmonar Obstructiva Crónica/fisiopatología , Pruebas de Función Respiratoria , Resultado del Tratamiento , AguaRESUMEN
CONCLUSIONS: The results reported here provide the first evidence of the production of superoxide, a biologically relevant reactive oxygen species (ROS), in human inner ear perilymph (hIP) in pathological conditions, by the activity of the xanthine dehydrogenase/xanthine oxidase (XA/XO) enzyme system. OBJECTIVE: To investigate the presence of ROS in hIP. METHODS: Since damage and apoptosis of inner ear hair cells may occur as a result of ROS-mediated injury, we investigated the presence and production of ROS in 105 hIP samples; 98 collected from patients affected by profound sensorineural hearing loss, during surgery for cochlear implantation, and 7 controls, collected from patients affected by otosclerosis, in case of spontaneous leakage after stapedotomy. ROS production was investigated by spectrophotometric analysis and polyacrylamide gel electrophoresis (SDS-PAGE). RESULTS: In hIP samples tested by cytochrome c reduction kinetics, the average superoxide production was 27.34 mumoles per mg of total protein, against 0.36 in controls. Some of these hIP samples, analyzed by cytochrome c reduction kinetics in the presence of xanthine, were found to be positive for ROS-producing XA/XO enzyme. These results were supported by SDS-PAGE analysis.
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Oído Interno/metabolismo , Oído Interno/patología , Perilinfa/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Adolescente , Adulto , Anciano , Niño , Preescolar , Implantes Cocleares , Citocromos c , Electroforesis en Gel de Poliacrilamida , Femenino , Pérdida Auditiva Sensorineural/diagnóstico , Pérdida Auditiva Sensorineural/metabolismo , Humanos , Lactante , Recién Nacido , Masculino , Persona de Mediana Edad , Otosclerosis/metabolismo , Espectrofotometría , Cirugía del Estribo , Ácido Úrico/metabolismo , Adulto JovenRESUMEN
Rhinoviruses are the major cause of the common cold and acute exacerbations of asthma and chronic obstructive pulmonary disease. We previously reported rapid rhinovirus induction of intracellular superoxide anion, resulting in NF-kappaB activation and pro-inflammatory molecule production. The mechanisms of rhinovirus superoxide induction are poorly understood. Here we found that the proteolytic activation of the xanthine dehydrogenase/xanthine oxidase (XD/XO) system was required because pretreatment with serine protease inhibitors abolished rhinovirus-induced superoxide generation in primary bronchial and A549 respiratory epithelial cells. These findings were confirmed by Western blotting analysis and by silencing experiments. Rhinovirus infection induced intracellular depletion of reduced glutathione (GSH) that was abolished by pretreatment with either XO inhibitor oxypurinol or serine protease inhibitors. Increasing intracellular GSH with exogenous H2S or GSH prevented both rhinovirus-mediated intracellular GSH depletion and rhinovirus-induced superoxide production. We propose that rhinovirus infection proteolytically activates XO initiating a pro-inflammatory vicious circle driven by virus-induced depletion of intracellular reducing power. Inhibition of these pathways has therapeutic potential.
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Células Epiteliales/metabolismo , Pulmón/metabolismo , Xantina Deshidrogenasa/metabolismo , Xantina Oxidasa/metabolismo , Línea Celular Tumoral , Activación Enzimática , Regulación Enzimológica de la Expresión Génica , Glutatión/metabolismo , Células HeLa , Humanos , Inflamación , Cinética , Modelos Biológicos , NADPH Oxidasas/metabolismo , Infecciones por Picornaviridae/metabolismo , Ácido Úrico/químicaRESUMEN
Suspension cultured cells of six rice cultivars differing in their sensitivity to blast were treated with mycelial wall hydrolysates prepared from seven isolates belonging to different Pyricularia grisea lineages. Soon after elicitor addition, rice cells produced significant amounts of superoxide anion, which was rapidly converted into diffusible peroxide. Maximal effects were achieved at 50 mg L-1 elicitor. In all cases, a 7 to 13-fold increase in the basal rate of reactive oxygen species production was found. Neither differential effects among strains nor clear relationships between lineage and the resulting oxidative burst were evident. Interestingly, a good correlation was found between basal (and elicited) levels of peroxide generation and the overall tolerance of rice cultivars to the pathogen. About two days after elicitation, cell death occurred proportional to the amount of hydrogen peroxide released. Peroxide was required to trigger loss of cell viability, but the latter was not due to a direct toxic effect, suggesting the induction of programmed cell death. Results represent the first data aimed to develop in vitro tests for pathogenicity prediction of Italian blast lineages toward rice cultivars.
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Ascomicetos/patogenicidad , Oryza/metabolismo , Oryza/microbiología , Apoptosis , Ascomicetos/aislamiento & purificación , Células Cultivadas , Peróxido de Hidrógeno/metabolismo , Italia , Oryza/citología , Enfermedades de las Plantas/microbiología , Especies Reactivas de Oxígeno/metabolismo , Estallido RespiratorioRESUMEN
Rhinoviruses are the major cause of common colds and of asthma exacerbations. Intercellular adhesion molecule-1 (ICAM-1) has a central role in airway inflammation and is the receptor for 90% of rhinoviruses. Rhinovirus infection of airway epithelium induces ICAM-1. Because redox state is directly implicated in inflammatory responses via molecular signaling mechanisms, here we studied the effects of reducing agents on rhinovirus-induced ICAM-1 expression, mRNA up-regulation, promoter activation, and nuclear factor activation. To investigate the effects of rhinovirus infection on the intracellular redox balance, we also studied whether rhinovirus infection triggers the production of reactive oxygen species. We found that reduced (GSH) but not oxidized (GSSG) glutathione (1-100 microM) inhibited in a dose-dependent manner rhinovirus-induced ICAM-1 up-regulation and mRNA induction in primary bronchial and A549 respiratory epithelial cells. GSH but not GSSG also inhibited rhinovirus-induced ICAM-1 promoter activation and rhinovirus-induced NF-kB activation. In parallel, we found that rhinovirus infection induced a rapid increase of intracellular superoxide anion that was maximal at the time of NF-kB activation. This oxidant generation was completely inhibited by GSH. We conclude that redox-mediated intracellular pathways represent an important target for the therapeutic control of rhinovirus-induced diseases.