Hsp90 beta inhibition modulates nitric oxide production and nitric oxide-induced apoptosis in human chondrocytes

Background: Hsp90 beta is a member of the Hsp90 family of protein chaperones. This family plays essential roles in the folding, maturation and activity of many proteins that are involved in signal transduction and transcriptional regulation. The role of this protein in chondrocytes is not well understood, although its increase in osteoarthritic cells has been reported. The present study aimed to explore the role of Hsp90 beta in key aspects of OA pathogenesis. Methods: Human OA chondrocytes were isolated from cartilage obtained from patients undergoing joint replacement surgery, and primary cultured. Cells were stimulated with proinflammatory cytokines (IL-1 beta or TNF-alpha) and nitric oxide donors (NOC-12 or SNP). For Hsp90 beta inhibition, two different chemical inhibitors (Geldanamycin and Novobiocin) were employed, or siRNA transfection procedures were carried out. Gene expression was determined by real-time PCR, apoptosis was quantified by flow cytometry and ELISA, and nitric oxide (NO) production was evaluated by the Griess method. Indirect immunofluorescence assays were performed to evaluate the presence of Hsp90 beta in stimulated cells. Results: Hsp90 beta was found to be increased by proinflammatory cytokines. Inhibition of Hsp90 beta by the chemicals Geldanamycin (GA) and Novobiocin (NB) caused a dose-dependent decrease of the NO production induced by IL-1 beta in chondrocytes, up to basal levels. Immunofluorescence analyses demonstrate that the NO donors NOC-12 and SNP also increased Hsp90 beta. Chemical inhibition or specific gene silencing of this chaperone reduced the DNA condensation and fragmentation, typical of death by apoptosis, that is induced by NO donors in chondrocytes. Conclusions: The present results show how Hsp90 beta modulates NO production and NO-mediated cellular death in human OA chondrocytes

Assessment of global DNA methylation in peripheral blood cell subpopulations of early rheumatoid arthritis before and after methotrexate

INTRODUCTION: DNA methylation is an epigenetic mechanism regulating gene expression that has been insufficiently studied in the blood of rheumatoid arthritis (RA) patients, as only T cells and total peripheral blood mononuclear cells (PBMCs) from patients with established RA have been studied and with conflicting results. METHOD: Five major blood cell subpopulations: T, B and NK cells, monocytes, and polymorphonuclear leukocytes, were isolated from 19 early RA patients and 17 healthy controls. Patient samples were taken before and 1 month after the start of treatment with methotrexate (MTX). Analysis included DNA methylation with high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry-selected reaction monitoring (HPLC-ESI-MS/MS-SRM) and expression levels of seven methylation-specific enzymes by quantitative polymerase chain reaction (qPCR). RESULTS: Disease-modifying anti-rheumatic drug (DMARD)-naive early RA patients showed global DNA hypomethylation in T cells and monocytes, together with a lower expression of DNA methyltrasnferase 1 (DNMT1), the maintenance DNA methyltransferase, which was also decreased in B cells. Furthermore, significantly increased expression of ten-eleven translocation1 (TET1), TET2 and TET3, enzymes involved in demethylation, was found in monocytes and of TET2 in T cells. There was also modest decreased expression of DNMT3A in B cells and of growth arrest and DNA-damage-inducible protein 45A (GADD45A) in T and B cells. Treatment with MTX reverted hypomethylation in T cells and monocytes, which were no longer different from controls, and increased global methylation in B cells. In addition, DNMT1 and DNMT3A showed a trend to reversion of their decreased expression. CONCLUSIONS: Our results confirm global DNA hypomethylation in patients with RA with specificity for some blood cell subpopulations and their reversal with methotrexate treatment. These changes are accompanied by parallel changes in the levels of enzymes involved in methylation, suggesting the possibility of regulation at this level

Regulated transcription of human matrix metalloproteinase 13 (MMP13) and interleukin-1β (IL1B) genes in chondrocytes depends on methylation of specific proximal promoter CpG sites

The role of DNA methylation in the regulation of catabolic genes such as MMP13 and IL1B, which have sparse CpG islands, is poorly understood in the context of musculoskeletal diseases. We report that demethylation of specific CpG sites at -110 bp and -299 bp of the proximal MMP13 and IL1B promoters, respectively, detected by in situ methylation analysis of chondrocytes obtained directly from human cartilage, strongly correlated with higher levels of gene expression. The methylation status of these sites had a significant impact on promoter activities in chondrocytes, as revealed in transfection experiments with site-directed CpG mutants in a CpG-free luciferase reporter. Methylation of the -110 and -299 CpG sites, which reside within a hypoxia-inducible factor (HIF) consensus motif in the respective MMP13 and IL1B promoters, produced the most marked suppression of their transcriptional activities. Methylation of the -110 bp CpG site in the MMP13 promoter inhibited its HIF-2alpha-driven transactivation and decreased HIF-2alpha binding to the MMP13 proximal promoter in chromatin immunoprecipitation assays. In contrast to HIF-2alpha, MMP13 transcriptional regulation by other positive (RUNX2, AP-1, ELF3) and negative (Sp1, GATA1, and USF1) factors was not affected by methylation status. However, unlike the MMP13 promoter, IL1B was not susceptible to HIF-2alpha transactivation, indicating that the -299 CpG site in the IL1B promoter must interact with other transcription factors to modulate IL1B transcriptional activity. Taken together, our data reveal that the methylation of different CpG sites in the proximal promoters of the human MMP13 and IL1B genes modulates their transcription by distinct mechanisms

On the front labelling of food: nutritional traffic lights, Nutri-Score and others

The labelling of packaged foods is a universal concern present in the national legislation of most countries. Regulation (EU) No. 1169/20111 on food information provided to the consumer allows the possibility of using a front nutrition label FOPL (Front-of-Pack nutrition label) in a complementary way to the mandatory nutrition information, on a voluntary basis, without replacing it, as long as the requirements mentioned in said Regulation are met, do not mislead the consumer are not ambiguous or confused and are based on relevant scientific data.Peer reviewe

The front labelling of food: Nutritional traffic lights, nutri-score and others

The labelling of packaged foods is a universal concern present in the national legislation of most countries. Regulation (EU) No. 1169/20112 on food information provided to the consumer allows the possibility of using a front nutrition label FOPL (Front-of-Pack nutrition label) in a complementary way to the mandatory nutrition information, on a voluntary basis, without replacing it, as long as the requirements mentioned in said Regulation are met, do not mislead the consumer, are not ambiguous or confusing and are based on relevant scientific data. The application of a "front" nutritional label is interesting in principle because it is more visible, unlike the mandatory nutritional label, which is located on the back or side of the packages. However, on the other hand, it can mislead the consumer should they intend to value the product nutritionally apart from the diet as a whole. An effective policy for the health of the citizen must be based on adequate training in food and consumption, starting from school age and reaching to society in general, contemplating the insertion of the variety of products in the variety of possible diets, according to the nutritional needs of the citizen, based on age, sex, lifestyle and sustainability. In this context, front labelling must be integrated into a global strategy to be effective and avoid being counterproductive. This document aims to offer food for thought to people, institutions and companies that have tomake decisions regarding food labelling.Peer reviewe

Prevalence of rhinitis and asthma respiratory symptoms in Spanish working population

Licencia para reproducir, transmitir y exhibir los documentos para cualquier propósito responsable sin fines de lucro, con la condición del reconocimiento apropiado de los créditos a la revista.[ES] Introducción: en salud laboral interesa conocer la prevalencia de los síntomas respiratorios relacionados con la rinitis y el asma en trabajadores españoles y sus variaciones por factores demográfi cos y laborales. El objetivo de esta investigación fue interrelacionar los síntomas respiratorios de vías altas y bajas. Métodos: se solicitó a 2603 trabajadores de distintos sectores y provincias españolas contestar una encuesta que fue supervisada por un médico del trabajo. Resultados: 438 trabajadores presentaron síntomas nasales crónicos (16.9 %) y 193 (7.5 %), síntomas pulmonares. De los 438 con síntomas nasales, 124 (28.3 %) presentaron síntomas pulmonares. Se observaron variaciones en los síntomas nasales por sexo, sector profesional, área de trabajo, tabaquismo y variaciones en los síntomas pulmonares por sector laboral, con mayor prevalencia en el sanitario, postal y transporte, en comparación con los sectores de la construcción, mueble o el metal. Conclusiones: la prevalencia de los síntomas nasales en la población trabajadora fue más elevada que la de los síntomas pulmonares. Los trabajadores afectados estaban infratratados, a pesar de padecer síntomas. Se observó interrelación entre los síntomas pulmonares y nasales de los trabajadores afectados.[EN] Objective: the aims were to examine the prevalence of respiratory symptoms associated with rhinitis and asthma in Spanish workers and its variations by age, sex, work sector, location in the workplace, smok- ing habits and the relationship between symptoms in the upper and lower respiratory tract. Methods: the sample was of 2603 workers in different sectors and regions throughout the country. They agreed and answered the ques- tions regarding respiratory symptoms, with the supervision of the com- pany s physician. Results: chronic nasal symptoms were present in 438 workers (16.9 %); 193 (7.5 %) had pulmonary symptoms. With nasal and pulmonary symp- toms were 124 patients (28.3 %). Likewise, of the 190 workers with pulmonary symptoms, 124 (64.2 %) had also nasal manifestations. We observed variations in nasal symptoms by sex, professional sector, work area and smoking habits, and variations in pulmonary symptoms by work sector. A higher prevalence was noted in the healthcare, postal, and transport sector as opposed to the construction, furniture or metal sector. Conclusions: the prevalence of nasal symptoms was higher than that of pulmonary symptoms in the working population. The workers affected by these pathologies were not treated adequately, despite they were pre- senting symptoms. We observed a relationship between pulmonary and nasal symptomsVicente Herrero, MT.; Prieto Andrés, L.; López González, ÁA.; Pérez Francés, C.; Ramirez Iñiguez De La Torre, MV.; Santamaria Navarro, C.; Terradillos García, MJ.... (2014). Síntomas respiratorios de rinitis y asma en población laboral española. Revista Medica- Instituto Mexicano Del Seguro Social. 52(1):50-59. http://hdl.handle.net/10251/62647S505952

Declaración de Chinchón: decálogo sobre eldulcorantes sin y bajos en calorías (ESBC)

Multidisciplinary experts in the areas of nutrition and health met in Chinchón, Madrid, on November 25-26, 2013 under the auspices of the Fundación para la Investigación Nutricional (Nutrition Research Foundation) and with the collaboration of the Madrid Regional Government’s Health Ministry, the International Sweeteners Association and the Carlos III Health Institute CIBER of Physiopathology of Obesity and Nutrition. They analyzed the current status of scientific knowledge on low- and no-calorie sweeteners (LNCS) and developed a consensus Decalogue on their use; this constitutes the Chinchón Declaration. Sweeteners, including sugar, represent a subject of undeniable interest and are currently a popular topic, although areas relating to their safety and benefits remain unknown to segments of academia and the general public. The nature of LNCS makes them vulnerable to biased and even contradictory information. They are food additives that are broadly used as sugar substitutes to sweeten foods, medicines and food supplements when non-nutritional or non-caloric alternatives are needed. The Chinchón Decalogue is the outcome of a meeting for reflection and consensus by a group of experts with backgrounds in different scientific disciplines (toxicology, clinical nutrition, community nutrition, physiology, food science, public health, pediatrics, endocrinology and nutrition, nursing, pharmaceutical care and food legislation). The Decalogue includes different aspects of LNCS related to regulation, use, benefits and safety. In general, benefits of LNCS have been traditionally neglected in comparison with the tendency for emphasising unexisting or unproven possible risks. The need to strengthen research on LNCS in Spain was emphasized, as well as the need to educate both professionals and the publicExpertos de carácter multidisciplinar de las áreas de conocimiento de la nutrición y la salud reunidos en Chinchón, Madrid, los días 25 y 26 de noviembre de 2013 , bajo los auspicios de la Fundación para la Investigación Nutricional y con la colaboración de la Consejería de Sanidad del Gobierno de la Comunidad de Madrid, la International Sweeteners Association y el CIBER de Fisiopatología de la Obesidad y la Nutrición del Instituto de Salud Carlos III, analizaron el estado actual del conocimiento científico en torno a los Edulcorantes sin y bajos en calorías (ESBC) y desarrollaron un Decálogo sobre su uso que constituye la Declaración de Chinchón. Los edulcorantes, incluido el azúcar, constituyen un elemento de indudable interés y actualidad, aunque no exento de desconocimiento por algunos sectores tanto académicos como de la población en general. La propia naturaleza de los ESBC los hace susceptibles de informaciones tergiversadas e incluso contradictorias. Son aditivos alimentarios ampliamente utilizados como sustitutivos del azúcar para endulzar alimentos, medicamentos y complementos alimenticios cuando se persiguen fines no nutritivos. El Decálogo de Chinchón es fruto de una reunión de reflexión y consenso por parte de un grupo de expertos procedentes de distintas disciplinas científicas (toxicología, nutrición clínica, nutrición comunitaria, fisiología, bromatología, salud pública, atención primaria, pediatría, endocrinología y nutrición, enfermería, atención farmacéutica y legislación alimentaria). El decálogo incluye diferentes aspectos de los EBSC relacionados con la legislación, uso, beneficios y seguridad. En general, los beneficios de los EBSC han sido tradicionalmente desatendidos en comparación con la tendencia de destacar posibles riesgos inexistentes o que no han sido probados. Hace especial hincapié en la necesidad de fortalecer la investigación de los EBSC en España, así como la necesidad de formar en este ámbito a los profesionales y a los consumidores en genera

Perspectivas actuales de los sujetos de derecho

En este volumen se recogen las intervenciones del II Seminario Internacional Permanente del Departamento de Derecho Internacional, Eclesiástico y Filosofía del Derecho de la Universidad Carlos III de Madrid que se desarrolló en el curso académico 2010-2011Presentación / Rafael de Asís Roig. - Algunas reflexiones sobre el individuo en Derecho Internacional / Castor Díaz Barrado. - Sobre capacidad y derechos / Rafael de Asís Roig. - Derecho Común versus Derecho Especial : el individuo como sujeto del derecho de libertad de conciencia y religiosa en España y Portugal / Alejandro Torres Gutiérrez. - Las empresas transnacionales como actores y sujetos «potenciales» en la sociedad internacional / Romualdo Bermejo García. - Las minorías religiosas como sujetos de derechos en España : un sistema de desigualdad / José María Contreras Mazario. - Los colectivos como sujetos de Derecho / Olga Sánchez Martínez. - De «protectorado internacional» a Estado «protegido» (¿Es Kosovo un Estado?) / Cesáreo Gutiérrez Espada - Contra el Derecho Internacional : imperialismo americano y subjetividad jurídica / Cristina García Pascual. - Cooperación constitucional e internacionalidad de la Iglesia católica / Paulino Pardo Prieto. - El individuo como titular de la libertad religiosa : aproximación a un problema de filosofía jurídica / Fernando Arlettaz. - La adopción de un procedimiento de comunicaciones individuales : ¿asignatura pendiente en la consolidación de la «subjetividad internacional del niño»? / Mª del Rosario Carmona Luque. - La persona con discapacidad como sujeto de derechos : análisis desde dos modelos / Patricia Cuenca Gómez. - Los sujetos de Derecho Internacional y los desastres internacionales / Carlos R. Fernández Liesa. - Los inmigrantes como colectivo : ¿son sujeto de Derecho Internacional? / Rosana Garciandía Garmendia. - La empresa : ¿Sujeto de Derecho Internacional? : Importancia de la cuestión / Hilda Garrido Suárez. - La tolerancia étnica y religiosa en los proyectos para la independencia del Estado búlgaro de la dominación otomana, el punto de partida del concepto legal sobre la protección de los derechos fundamentales de las minorias en Bulgaria / Ángel Hristov Kolev. - El régimen jurídico del profesorado y de la asignatura de religión católica en la escuela pública. Comentario a la STC 51/2011 de 14 de abril / Andrés Murcia González. - El sujeto de derecho en Pufendorf / Antonio Pele. - ¿Puede un sujeto de Derecho Internacional juzgar a otro? Algunas consideraciones sobre la inmunidad de jurisdicción de las organizaciones internacionales / Juan Jorge Piernas Lópe

Derecho y tecnologías avanzadas

¿Están reñidos el Derecho y las Administraciones Públicas con las tecno- logías más avanzadas de la información y la comunicación? La respuesta negativa a esta pregunta fue la conclusión de las aportaciones de los participantes en las IV Jornadas sobre Derecho y Tecnología y en el XIV Encuentro Ibero-Latino-Americano sobre Gobierno Electrónico e Inclusión Digital, celebrados en Zaragoza los días 18 y 19 de junio de 2012. Cada uno, desde su perspectiva y campo de actividad, trató de aportar pruebas de cómo las tecnologías de la información y la comunicación más avanzadas aportan soluciones en el campo del Derecho y las Admi- nistraciones Públicas. Estas aportaciones están recogidas en este libro, agrupadas en tres blo- ques diferentes. En el primero, bajo el título Tecnologías avanzadas y Derecho se encuen- tran los trabajos aportados sobre el uso de tecnologías avanzadas en actividades relacionadas con el Derecho y la Administración de Justicia, con casos concretos de Brasil y España, o de tipo general, aplicable en cualquier país. En el segundo, el título Administración electrónica: acceso de los ciudadanos a los servicios públicos reúne experiencias concretas llevadas a cabo en España, Unión Europea y Brasil. Por último, y no menos importante, el título Aprendizaje y tecnologías recoge propuestas para fomentar el uso de las nuevas tecnologías en la docencia, tanto en Derecho como en otros campos de la docencia universitaria