IL-10 deficiency exacerbates the brain inflammatory response to permanent ischemia without preventing resolution of the lesion

El pdf del artículo es la versión post-print.Stroke induces inflammation that can aggravate brain damage. This work examines whether interleukin-10 (IL-10) deficiency exacerbates inflammation and worsens the outcome of permanent middle cerebral artery occlusion (pMCAO). Expression of IL-10 and IL-10 receptor (IL-10R) increased after ischemia. From day 4, reactive astrocytes showed strong IL-10R immunoreactivity. Interleukin-10 knockout (IL-10 KO) mice kept in conventional housing showed more mortality after pMCAO than the wild type (WT). This effect was associated with the presence of signs of colitis in the IL-10 KO mice, suggesting that ongoing systemic inflammation was a confounding factor. In a pathogen-free environment, IL-10 deficiency slightly increased infarct volume and neurologic deficits. Induction of proinflammatory molecules in the IL-10 KO brain was similar to that in the WT 6 hours after ischemia, but was higher at day 4, while differences decreased at day 7. Deficiency of IL-10 promoted the presence of more mature phagocytic cells in the ischemic tissue, and enhanced the expression of M2 markers and the T-cell inhibitory molecule CTLA-4. These findings agree with a role of IL-10 in attenuating local inflammatory reactions, but do not support an essential function of IL-10 in lesion resolution. Upregulation of alternative immunosuppressive molecules after brain ischemia can compensate, at least in part, the absence of IL-10. © 2013 ISCBFM.Work supported by the Spanish Ministry of Economy (SAF2011-30492), and the European Community (FP7, grant agreements: n°201024 ARISE and n°278850 InMiND), and the ERANET-NEURON project (PRI-PIMNEU-2011-1342). IPP and EBT had PhD fellowships from the Agència de Gestió d'Ajuts Universitaris i de Recerca (AGAUR) of the Generalitat de Catalunya and the FPU program of the Spanish Ministry of Economy, respectively.Peer Reviewe

FGFR1 Cooperates with EGFR in Lung Cancer Oncogenesis, and Their Combined Inhibition Shows Improved Efficacy

Introduction: There is substantial evidence for the onco- genic effects of fi broblast growth factor receptor 1 (FGFR1) in many types of cancer, including lung cancer, but the role of this receptor has not been addressed speci fi cally in lung adenocarcinoma. Methods: We performed FGFR1 and EGFR overexpression and co-overexpression assays in adenocarcinoma and in inmortalized lung cell lines, and we also carried out surrogateandinteractionassays.Weperformedmono- therapy and combination EGFR /FGFR inhibitor sensitivity assays in vitro and in vivo in cell line – and patient- derived xenografts. We determined FGFR1 mRNA expression in a cohort of patients with anti – EGFR ther- apy – treated adenocarcinoma. Results: We have reported a cooperative interaction between FGFR1 and EGFR in this context, resulting in increased EGFR activation and oncogenic signaling. We have provided in vitro and in vivo evidence indicating that FGFR1 expression in- creases tumorigenicity in cells with high EGFR activation in EGFR-mutated and EGFR wild-type models. At the clinical level, we have shown that high FGFR1 expression levels pre- dict higher resistance to erlotinib or ge fi tinib in a cohort of patients with tyrosine kinase inhibitor – treated EGFR-mutated and EGFR wild-type lung adenocarcinoma. Dual EGFR and FGFR inhibition in FGFR1-over expressing, EGFR-activated models shows synergistic effects on tumor growth in vitro and in cell line – and patient-derived xenografts, suggesting that patients with tumors bearing these characteristics may bene fi t from combined EGFR/FGFR inhibition. Conclusion: These results support the extended the use of EGFR inhibitors beyond monotherapy in the EGFR-mutated adenocarcinoma setting in combination with FGFR in- hibitors for selected patients with increased FGFR1 over- expression and EGFR activation.ISCIII PI14/01964 PIE15/00076 PI17/00778 DTS17/00089 PI15/00045 PI17/00033 PI16/01311 FI12/00429CIBERONC CD16/12/00442FEDER CD16/12/00442 PI16/01311Spanish Ministry of Economy and Competitiveness PI15/00045Ministry of Health and Social Welfare of Junta de Andalucía PI-0046-2012 C-0040-2016Ministry of Equality, Health and Social Policies of the Junta de Andalucía PI- 0029-2013Comunidad de Madrid B2017/BMD3884Ministry of Education, Culture and Sports FPU13/0259

Controlled manipulation of enzyme specificity through immobilization-induced flexibility constraints

It is thought that during immobilization enzymes, as dynamic biomolecules, may become distorted and this may alter their catalytic properties. However, the effects of different immobilization strategies on enzyme rigidity or flexibility and their consequences in specificity and stereochemistry at large scale has not been yet clearly evaluated and understood. This was here investigated by using as model an ester hydrolase, isolated from a bacterium inhabiting a karstic lake, with broad substrate spectrum (72 esters being converted; 61.5 U mg−1 for glyceryl tripropionate) but initially non-enantiospecific. We found that the enzyme (7 nm × 4.4 nm × 4.2 nm) could be efficiently ionic exchanged inside the pores (9.3 nm under dry conditions) of amino-functionalized ordered mesoporous material (NH2-SBA-15), achieving a protein load of 48 mg g−1, and a specific activity of 4.5 ± 0.1 U mg−1. When the enzyme was site-directed immobilized through His interaction with an immobilized cationon the surface of two types of magnetic micro-particles through hexahistidine-tags, protein loads up to 10.2 μg g−1 and specific activities of up to 29.9 ± 0.3 U mg−1, were obtained. We found that ionically exchanged enzyme inside pores of NH2-SBA-15 drastically narrowed the substrate range (17 esters), to an extent much higher than ionically exchanged enzyme on the surface of magnetic micro-particles (up to 61 esters). This is attributed to differences in surface chemistry, particle size, and substrate accessibility to the active site tunnel. Our results also suggested, for the first time, that immobilization of enzymes in pores of similar size may alter the enzyme structures and produce enzyme active centers with different configuration which promote stereochemical conversions in a manner different to those arising from surface immobilization, where the strength of the ionic exchange also has an influence. This was shown by demonstrating that when the enzyme was introduced inside pores with a diameter (under dry conditions) slightly higher than that of the enzyme crystal structure a biocatalyst enantiospecific for ethyl (R)-4-chloro-3-hydroxybutyrate was produced, a feature not found when using wider pores. By contrast, immobilization on the surface of ferromagnetic microparticles produced selective biocatalysts for methyl (S)-(+)-mandelate or methyl (S)-lactate depending on the functionalization. This study illustrates the benefits of extensive analysis of the substrate spectra to better understand the effects of different immobilization strategies on enzyme flexibility/rigidity, as well as substrate specificity and stereochemistry. Our results will help to design tunable materials and interfaces for a controlled manipulation of specificity and to transform non-enantiospecific enzymes into stereo-chemically substrate promiscuous biocatalysts capable of converting multiple chiral molecules.This project received funding from the European Union’s Horizon 2020 research and innovation program Blue Growth: Unlockingthe potential of Seas and Oceans under grant agreement no. 634486 (project acronym INMARE). This research was also supported by the grants PCIN-2014-107 (within ERA NET IB2 grant nr. ERA-IB-14-030 - MetaCat), PCIN-2017-078 (within the ERA-MarineBiotech grant ProBone), BIO2014-54494-R, MAT2016-77496-R, BIO2017-85522-R, and CTQ2016-79138-R from the Spanish Ministry of Economy, Industry and Competitiveness. A.B. acknowledges the support from the Spanish Ministry of Economy, Industry and Competitiveness (MAT2017-88808-R grant), María de Maeztu Units of Excellence Programme (MDM-2016-0618), and the Diputación de Guipúzcoa for current funding in the frame of Gipuzkoa Fellows program. G.D. thanks the German Federal Ministry of Education and Research (BMBF, Grant No. 031A095C) for funding in the frame of the Molecular Interaction Engineering program (Biotechnologie 2020+). The authors gratefully acknowledge financial support provided by the European Regional Development Fund (ERDF). C.C. thanks the Spanish Ministry of Economy, Industry and Competitiveness for a PhD fellowship (Grant BES-2015-073829).Peer ReviewedPostprint (published version

Modulation of phenolic metabolism under stress conditions in a Lotus japonicus mutant lacking plastidic glutamine synthetase

This paper was aimed to investigate the possible implications of the lack of plastidic glutamine synthetase (GS2) in phenolic metabolism during stress responses in the model legume Lotus japonicus. Important changes in the transcriptome were detected in a GS2 mutant called Ljgln2-2, compared to the wild type, in response to two separate stress conditions, such as drought or the result of the impairment of the photorespiratory cycle. Detailed transcriptomic analysis showed that the biosynthesis of phenolic compounds was affected in the mutant plants in these two different types of stress situations. For this reason, the genes and metabolites related to this metabolic route were further investigated using a combined approach of gene expression analysis and metabolite profiling. A high induction of the expression of several genes for the biosynthesis of different branches of the phenolic biosynthetic pathway was detected by qRT-PCR. The extent of induction was always higher in Ljgln2-2, probably reflecting the higher stress levels present in this genotype. This was paralleled by accumulation of several kaempferol and quercetine glycosides, some of them described for the first time in L. japonicus, and of high levels of the isoflavonoid vestitol. The results obtained indicate that the absence of GS2 affects different aspects of phenolic metabolism in L. japonicus plants in response to stress.España FEDER-Ministerio de Economía y Competitividad AGL2014-54413-REspaña Junta de Andalucía,Consejería de Economía, Innovación y Ciencia, P1O-CVI-6368España Junta de Andalucía,Consejería de Economía, Innovación y Ciencia, BIO-16

Definición de una guía metodológica de gestión de proyectos basado en PMBOK V6 para el departamento de mantenimiento de producción de El Tiempo Casa Editorial.

Guía estandarizada, Formatos de gestión de proyectos.Ante la actual situación coyuntural que se vive en la empresa y en general en el gremio de productos impresos, evoca al mejoramiento de los procesos a cada uno de los departamentos que conforman la organización en aras de optimizar los recursos asignados, por tal razón, la principal motivación en de la realización de la presente propuesta es la de redirigir esfuerzo para ser más eficiente a la hora de formular y gestionar los proyectos que se desarrollan en el área de Mantenimiento de producción. La propuesta en sí constituye en la definición de una guía estándar metodológica basada en el PMBOK para tal fin, analizando en primera instancia la metodología actual, identificando sus posibles mejoras y adaptándola a las diferentes procesos y área de conocimiento planteados por el PMI dentro del PMBOK V6. Como resultado de la investigación se diseñaron dos formatos, una guía detallada en Word y formularios en Excel, ajustados a las necesidades del área de negocio y a las directrices de la compañía

DESARROLLO DE LAS ACCIONES MOTRICES EN EL MEDIO NATURAL EN LOS DISTINTOS CENTROS DE HUESCA

En el área de Educación Física de la Educación Secundaria Obligatoria en la Comunidad Autónoma de Aragón, están incluidos 6 bloques de contenidos, dentro de los cuales se encuentra el bloque 4, que recoge las Acciones motrices en el medio natural. El presente trabajo pretende realizar un análisis sobre cómo se desarrolla este bloque en los distintos centros de Huesca. Se va a enviar un cuestionario a los profesores de Educación Física para analizar en qué medida se expone dicho bloque de contenidos y cuáles son las actividades que se realizan a lo largo del curso. Los resultados muestran que un 88% incluye este bloque de contenidos en su programación, pese a la cantidad de limitaciones que encuentran. Destacando así la gran importancia de los beneficios que aportan a quienes practican Acciones motrices en el medio natural. <br /

Identificación de barreras y facilitadores para la (des)prescripción de benzodiacepinas: un estudio cualitativo con pacientes y profesionales sanitarios

Background. There has been a steadily growing trend in prescribing benzodiazepines over last decade. Spain is one of the countries where this class of drugs is most extensively prescribed by primary healthcare physicians. The aim of this study is to identify factors that might be acting as barriers and enablers for benzodiazepine (de)prescription from patient and professional perspectives. Methods. Qualitative study through semi-structured interviews with medical practitioners (n=17) and patients (n=27), and a nominal group with medical practitioners (n=19). Interviews were audio-recorded, transcribed and analyzed using thematic analysis. Results. The analysis revealed key themes and was organized around barriers and enablers connected to three interrelated dimensions: the social and community context of prescription; the structure, organization and/or management of the health system, and the doctor-patient relationship. The excessive workload of professionals was widely cited as influencing over-prescription. (De) prescription of benzodiazepine was facilitated by encouraging the social prescription of health assets or developing strategies to therapeutic alliance processes and better doctor-patient communication. Conclusion. Our findings suggest that there is a role for the salutogenic approach and the health asset model in the development of a more person-centred clinical care. This study considers the importance of encouraging the use of non-pharmacological methods and techniques in the health system and promoting the creation of multidisciplinary teams, therapeutic alliance processes and better doctor-patient communication by giving professionals training in psychosocial skills.Fundamento. La tendencia en la prescripción de benzodiacepinas ha crecido en la última década. España está entre los países donde este tipo de fármacos es el más prescrito por profesionales en Atención Primaria. El propósito de este estudio es identificar factores que podrían estar actuando como barreras y facilitadores en la (des)prescripción de benzodiacepinas desde la perspectiva de pacientes y profesionales sanitarios. Material y métodos. Estudio cualitativo a través de entrevistas semiestructuradas con profesionales sanitarios (n=17) y pacientes (n=27), y un grupo nominal con profesionales sanitarios (n=19). Las entrevistas fueron transcritas y analizadas utilizando un análisis temático. Resultados. El análisis reveló temas claves organizados como barreras y facilitadores conectados a tres dimensiones interrelacionadas: el contexto comunitario y social de la prescripción; la estructura, organización y/o gestión del sistema sanitaria, y la relación médico-paciente. La excesiva carga laboral de los profesionales fue ampliamente citada como influyente en la prescripción excesiva. Acciones como promover la prescripción social de activos en salud o desarrollar estrategias para facilitar la alianza terapéutica y mejorar la comunicación médico-paciente, fueron vistos como facilitadores. Conclusiones. Los hallazgos sugieren el rol que el enfoque salutogénico y el modelo de activos en salud pueden jugar en el desarrollo de una atención clínica centrada en la persona. El estudio considera la importancia de promover métodos y técnicas de intervención no farmacológicos, la promoción de equipos multidisciplinares y la formación en habilidades psicosociales.This work was supported by the CIBER of Epidemiology and Public Health (CIBERESP)

Lack of validation of genetic variants associated with anti-tumor necrosis factor therapy response in rheumatoid arthritis: a genome-wide association study replication and meta-analysis

Introduction: In this study, our aim was to elucidate the role of four polymorphisms identified in a prior large genome-wide association study (GWAS) in which the investigators analyzed the responses of patients with rheumatoid arthritis (RA) to treatment with tumor necrosis factor inhibitors (TNFi). The authors of that study reported that the four genetic variants were significantly associated. However, none of the associations reached GWAS significance, and two subsequent studies failed to replicate these associations. Methods: The four polymorphisms (rs12081765, rs1532269, rs17301249 and rs7305646) were genotyped in a total of 634 TNFi-treated RA patients of Spanish Caucasian origin. Four outcomes were evaluated: changes in the Disease Activity Score in 28 joints (DAS28) after 6 and 12 months of treatment and classification according to the European League Against Rheumatism (EULAR) response criteria at the same time points. Association with DAS28 changes was assessed by linear regression using an additive genetic model. Contingency tables of genotype and allele frequencies between EULAR responder and nonresponder patients were compared. In addition, we combined our data with those of previously reported studies in a meta-analysis including 2,998 RA patients. Results: None of the four genetic variants showed an association with response to TNFi in any of the four outcomes analyzed in our Spanish patients. In addition, only rs1532269 yielded a suggestive association (P = 0.0033) with the response to TNFi when available data from previous studies were combined in the meta-analysis. Conclusion: Our data suggest that the rs12081765, rs1532269, rs17301249 and rs7305646 genetic variants do not have a role as genetic predictors of TNFi treatment outcomes

Pancreatitis aguda grave en un paciente fumador

The case report of a 51 years patient with personal pathological history of being a heavy smoker is described, who went to the emergency room of Dr. Joaquín Castillo Duany Teaching Provincial Hospital in Santiago de Cuba due to a dyspnea associated with high figures of blood pressure and livedo reticularis in the abdominal anterior face. Due to the clinical pattern and the results of the complementary tests, he was referred to the Intensive Cares Unit, where later on he was diagnosed serious acute pancreatitis, after an exploratory laparotomy where ischemic areas in the intestinal loops were found. The patient maintained a torpid clinical course and he died because of a multiple dysfunction of organs.Se describe el caso clínico de un paciente de 51 años de edad con antecedente patológico personal de ser un fumador empedernido, el cual acudió al Cuerpo de Guardia del Hospital Provincial Docente Dr. Joaquín Castillo Duany de Santiago de Cuba por presentar disnea asociada a cifras elevadas de tensión arterial y livedo reticular en la cara anterior abdominal. Debido al cuadro clínico y a los resultados de los exámenes complementarios, fue trasladado a la Unidad de Cuidados Intensivos, donde posteriormente se le diagnosticó pancreatitis aguda grave, tras realizar una laparotomía exploratoria y hallar zonas isquémicas en las asas intestinales. El paciente mantuvo una evolución tórpida y falleció a causa de una disfunción múltiple de órganos