Medical ozone promotes Nrf2 phosphorylation reducing oxidative stress and pro-inflammatory cytokines in multiple sclerosis patients.

Oxidative stress and inflammation play key roles in the pathogenesis of Multiple sclerosis (MS). Different drugs have been used in the clinical practice, however, there is not a completely effective treatment. Due to its potential therapeutic action, medical ozone represents a promising approach for neurodegenerative disorders. The aim of the present study was to address the role of ozone therapy on the cellular redox state in MS patients. Ozone (20µg/ml) was administered three times per week during a month by rectal insufflation. The effect of ozone therapy on biomarkers of oxidative stress and inflammation was addressed by spectrophotometric and immunoenzymatic assays. Furthermore, we investigated the action of ozone on CK2 expression and Nrf2 phosphorylation by western blotting analysis. Medical ozone significantly improved (P < 0.05) the activity of antioxidant enzymes and increased the levels of cellular reduced glutathione. In accordance, a significant reduction (P < 0.05) of oxidative damage on lipids and proteins was observed in ozone-treated patients. As well, the levels of pro-inflammatory cytokines TNFα and IL-1ß were lower after ozone treatment. Ozone therapy incremented the CK2 expression together with Nrf2 phosphorylation in mononuclear cells of MS patients. These findings suggest that ozone´s antioxidant and anti-inflammatory effects might be partially associated with an induction of Nrf2 phosphorylation and activation. These results provide new insights on the molecular events modulated by ozone, and pointed out ozone therapy as a potential therapeutic alternative for MS patients.

Arresting progressive atherosclerosis by immunization with an anti-glycosaminoglycan monoclonal antibody in apolipoprotein E-deficient mice.

Atherogenesis is associated with the early retention of low-density lipoproteins (LDL) in the arterial intima by interaction with glycosaminoglycan (GAG)-side chains of proteoglycans. Retained LDL undergo reactive oxygen species-mediated oxidation. Oxidized LDL trigger oxidative stress (OS) and inflammation, contributing to atherosclerosis development. Recently, we reported the preventive anti-atherogenic properties of the chimeric mouse/human monoclonal antibody (mAb) chP3R99-LALA, which were related to the induction of anti-chondroitin sulfate antibody response able to inhibit chondroitin sulfate dependent LDL-enhanced oxidation. In the present work, we aimed at further investigating the impact of chP3R99-LALA mAb vaccination on progressive atherosclerosis in apolipoprotein E-deficient mice (apoE(-/-)) fed with a high-fat high-cholesterol diet receiving 5 doses (50 µg) of the antibody subcutaneously, when ~5% of the aortic area was covered by lesions. Therapeutic immunization with chP3R99-LALA mAb halted atherosclerotic lesions progression. In addition, aortic OS was modulated, as shown by a significant (p<0.05) reduction of lipid and protein oxidation, preservation of antioxidant enzymes activity and reduced glutathione, together with a decrease of nitric oxide levels. chP3R99-LALA mAb immunization also regulated aortic NF-κB activation, diminishing the proinflammatory IL1-ß and TNF-α gene expression as well as the infiltration of macrophages into the arterial wall. The therapeutic immunization of apoE(-/-) with progressive atheromas and persistent hypercholesterolemia using chP3R99-LALA mAb arrested further development of lesions, accompanied by a decrease of aortic OS and NF-κB-regulated pro-inflammatory cytokine gene expression. These results contribute to broaden the potential use of this anti-GAG antibody-based immunotherapy as a novel approach to target atherosclerosis at different phases of progression.

Ozone-Oxidative Preconditioning Prevents Doxorubicin-induced Cardiotoxicity in Sprague-Dawley Rats.

OBJECTIVES: Induced dilated cardiomyopathy is the main limitation of the anti-cancer drug doxorubicin, which causes oxidative stress and cardiomyocyte death. As ozone therapy can activate the antioxidant systems, this study aimed to investigate the therapeutic efficacy of ozone-oxidative preconditioning against doxorubicin-induced cardiotoxicity. METHODS: The study was carried out from September 2013 to January 2014. Sprague-Dawley rats were randomly distributed in the following treatment groups: Group 1 were treated with 2 mg/kg intraperitoneal (i.p.) of doxorubicin twice a week for 50 days; Group 2 were treated with 0.3 mg of ozone/oxygen mixture at 50 µg/mL of ozone per 6 mL of oxygen by rectal insufflation and then treated with doxorubicin; Group 3 were treated as Group 2 but only with the oxygen, and Group 4 were treated with oxygen first, and then with sodium chloride i.p. as the control group. RESULTS: The results showed that ozone therapy preserved left ventricle morphology which was accompanied by a reduction of serum pro-brain natriuretic peptide levels. The cardioprotective effects of ozone-oxidative preconditioning were associated with a significant increase (P <0.05) of antioxidant enzymes activities and a reduction of lipid and protein oxidation (P <0.05). CONCLUSION: Ozone-oxidative preconditioning prevents doxorubicin-induced dilated cardiomyopathy through an increase of antioxidant enzymes and a reduction of oxidised macromolecules. This establishes the background for future studies to determine if ozone therapy can be used as a complementary treatment for attenuating doxorubicin-induced cardiotoxicity in cancer patients.

Diferencias del estado redox entre espermatozoides y plasma seminal en adultos jóvenes / Difference of redux status between spermatozoa and seminal plasma in young adults

Introducción: Las especies reactivas del oxígeno y el estrés oxidativo desempeñan un papel fundamental en la infertilidad masculina. Sin embargo, estas especies oxidantes también han sido asociadas con los procesos de capacitación de los gametos masculinos cuando son generadas a bajos niveles y de manera controlada. Actualmente los biomarcadores redox son introducidos en el diagnóstico clínico de la infertilidad masculina en el mundo, como una herramienta complementaria a los parámetros del espermiograma. Sin embargo, en Cuba, esta metodología aún no se encuentra extendida a los servicios de salud. Objetivo: El objetivo del presente trabajo fue caracterizar el estado redox de espermatozoides y el líquido seminal de sujetos aparentemente sanos, a través de la determinación de una serie de biomarcadores de estrés oxidativo. Los niveles de malonildialdehído, la actividad de las enzimas antioxidantes superóxido dismutasa y catalasa, así como los niveles de glutatión reducido, el potencial de peroxidación y la capacidad reductora fueron determinados mediante técnicas espectrofotométricas. Métodos: Se analizaron 40 muestras de semen de sujetos aparentemente sanos, las cuales fueron obtenidas mediante masturbación sin el empleo de lubricantes y con al menos tres días de abstinencia eyaculatoria. En el estudio se incluyeron sujetos de 20 a 35 años, aparentemente sanos según exámenes de laboratorio clínico y con paternidad probada. Resultados: Los resultados demostraron que existen diferencias significativas (p< 0,05) entre los marcadores evaluados en el líquido seminal con respecto al espermatozoide, lo cual sugiere la existencia de un estado redox diferenciado entre ambos compartimentos. Conclusiones: Estos hallazgos demuestran la necesidad de evaluar el nivel de estrés oxidativo tanto en las células sexuales como en el fluido que las contiene. Ello contribuirá, sin lugar a dudas, a un diagnóstico más eficaz e integral de la capacidad fértil del hombre

Background: Reactive oxygen species and oxidative stress play a fundamental role in male infertility. However, these oxidizing species have also been associated with the process of capability of male gametes when they are generated at low levels and in a controlled manner. Currently, redox biomarkers are introduced in the clinical diagnosis of male infertility in the world as a complementary tool to the spermiogram parameters. However, in Cuba, this methodology is not yet extended to health services. Objective: The objective of this study was to characterize the redox status of spermatozoa and seminal fluid of apparently healthy subjects through the identification of a number of biomarkers of oxidative stress. Methods: 40 samples of semen of apparently healthy subjects were analyzed, which were obtained by masturbation without the use of lubricants and with at least 3 days of ejaculatory abstinence. The study included subjects from 20 to 35 years of age, who were apparently healthy according to both laboratory tests and paternity test results. Results: The results showed that there are significant differences (p<0, 05) between the markers evaluated in the seminal fluid and the spermatozoon which suggests the existence of a differentiated redux status between the two compartments. Conclusions: These findings demonstrate the need to evaluate the level of oxidative stress in both sexual cells and the fluid that contains them. It will contribute, with no doubt, to a more effective and comprehensive diagnosis of man's fertility

Diferencias del estado redox entre espermatozoides y plasma seminal en adultos jóvenes / Difference of redux status between spermatozoa and seminal plasma in young adults

Introducción: Las especies reactivas del oxígeno y el estrés oxidativo desempeñan un papel fundamental en la infertilidad masculina. Sin embargo, estas especies oxidantes también han sido asociadas con los procesos de capacitación de los gametos masculinos cuando son generadas a bajos niveles y de manera controlada. Actualmente los biomarcadores redox son introducidos en el diagnóstico clínico de la infertilidad masculina en el mundo, como una herramienta complementaria a los parámetros del espermiograma. Sin embargo, en Cuba, esta metodología aún no se encuentra extendida a los servicios de salud. Objetivo: El objetivo del presente trabajo fue caracterizar el estado redox de espermatozoides y el líquido seminal de sujetos aparentemente sanos, a través de la determinación de una serie de biomarcadores de estrés oxidativo. Los niveles de malonildialdehído, la actividad de las enzimas antioxidantes superóxido dismutasa y catalasa, así como los niveles de glutatión reducido, el potencial de peroxidación y la capacidad reductora fueron determinados mediante técnicas espectrofotométricas. Métodos: Se analizaron 40 muestras de semen de sujetos aparentemente sanos, las cuales fueron obtenidas mediante masturbación sin el empleo de lubricantes y con al menos tres días de abstinencia eyaculatoria. En el estudio se incluyeron sujetos de 20 a 35 años, aparentemente sanos según exámenes de laboratorio clínico y con paternidad probada. Resultados: Los resultados demostraron que existen diferencias significativas (p< 0,05) entre los marcadores evaluados en el líquido seminal con respecto al espermatozoide, lo cual sugiere la existencia de un estado redox diferenciado entre ambos compartimentos. Conclusiones: Estos hallazgos demuestran la necesidad de evaluar el nivel de estrés oxidativo tanto en las células sexuales como en el fluido que las contiene. Ello contribuirá, sin lugar a dudas, a un diagnóstico más eficaz e integral de la capacidad fértil del hombre(AU)

Background: Reactive oxygen species and oxidative stress play a fundamental role in male infertility. However, these oxidizing species have also been associated with the process of capability of male gametes when they are generated at low levels and in a controlled manner. Currently, redox biomarkers are introduced in the clinical diagnosis of male infertility in the world as a complementary tool to the spermiogram parameters. However, in Cuba, this methodology is not yet extended to health services. Objective: The objective of this study was to characterize the redox status of spermatozoa and seminal fluid of apparently healthy subjects through the identification of a number of biomarkers of oxidative stress. Methods: 40 samples of semen of apparently healthy subjects were analyzed, which were obtained by masturbation without the use of lubricants and with at least 3 days of ejaculatory abstinence. The study included subjects from 20 to 35 years of age, who were apparently healthy according to both laboratory tests and paternity test results. Results: The results showed that there are significant differences (p<0, 05) between the markers evaluated in the seminal fluid and the spermatozoon which suggests the existence of a differentiated redux status between the two compartments. Conclusions: These findings demonstrate the need to evaluate the level of oxidative stress in both sexual cells and the fluid that contains them. It will contribute, with no doubt, to a more effective and comprehensive diagnosis of man's fertility(AU)