Inhibidores de las vías intracelulares NF-κB y JAK/STAT en el tratamiento experimental de la aterosclerosis y la nefropatía diabética

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Farmacología. Fecha de lectura: 10-07-2015La diabetes es una enfermedad crónica de gran impacto socio-sanitario debido a su elevada frecuencia y a las consecuencias de sus complicaciones crónicas. Entre ellas, la nefropatía y la aterosclerosis son principal causa de enfermedad renal crónica y morbimortalidad en los pacientes diabéticos, aunque en la actualidad no existen alternativas terapéuticas que frenen de forma efectiva la aparición de eventos cardiovasculares o la progresión a insuficiencia renal. La inflamación es un proceso clave en la patogenia del daño renal y vascular en la diabetes y en ella participan numerosas proteínas inflamatorias, cuya expresión está estrictamente regulada a nivel intracelular por diversos mecanismos. En esta tesis hemos estudiado el potencial terapéutico de dos vías intracelulares activadas durante el inicio y progresión de la nefropatía diabética y la aterosclerosis: NF-κB (factor nuclear-κB) y JAK/STAT (Janus quinasa/Transductor de la señal y activador de la transcripción). En la primera parte, analizamos las consecuencias funcionales de inhibir específicamente la ruta NF-κB dependiente de la IκB quinasa (IKK) en un modelo experimental de nefropatía y aterosclerosis asociadas a diabetes, inducido por inyección de estreptozotocina en ratones deficientes en apolipoproteína E (apoE-/-). Para ello, empleamos un péptido con permeabilidad celular y que bloquea la formación del complejo IKK. El tratamiento con el péptido inhibidor no afectó a la hiperglucemia ni al perfil lipídico de los ratones diabéticos pero mejoró su función renal, reduciendo significativamente las lesiones renales (expansión mesangial, fibrosis e infiltrado), la actividad NF-κB y la expresión de genes proinflamatorios y profibróticos. El péptido también suprimió la actividad NF-κB en la aorta de los ratones diabéticos, redujo el tamaño de las placas de ateroma y su contenido lipídico e inflamatorio (leucocitos y citoquinas) y aumentó los componentes de estabilización (colágeno y células de músculo liso vascular). In vitro, el péptido bloqueó la interacción entre las subunidades IKKγ e IKKα/β y previno la translocación nuclear de NF-κB y la expresión de genes controlados por este factor, así como la proliferación celular en condiciones de hiperglucemia e inflamación. En la segunda parte, investigamos el efecto ateroprotector de la inhibición de JAK/STAT en ratones apoE-/- mediante terapia génica con un adenovirus que codifica la proteína SOCS3 (supresora de la señal de citoquinas 3). La expresión del transgen SOCS3 atenuó la activación de STAT1 y STAT3 en aorta, redujo la expresión de sus genes dependientes (citoquinas, quimioquinas, moléculas de adhesión y receptores scavenger) y previno el desarrollo de aterosclerosis en fases tempranas. Además, la terapia con SOCS3 promovió un fenotipo de placa más estable, caracterizado por un menor contenido de lípidos, células T y macrófagos M1 y un incremento de macrófagos M2 y colágeno. Sin embargo, la expresión de SOCS3 no consiguió frenar la progresión de las lesiones ateroscleróticas en fases más avanzadas del modelo animal. A nivel sistémico, la terapia con SOCS3 redujo la expresión de citoquinas de respuesta linfocitaria Th1 y Th17 y el estado de activación de los monocitos circulantes. In vitro, SOCS3 inhibió la activación de STAT inducida por citoquinas en células vasculares y macrófagos, la expresión de genes proinflamatorios y la proliferación. En conclusión, nuestros estudios demuestran el efecto renoprotector y antiaterosclerótico de la inhibición selectiva de las vías NF-κB y JAK/STAT en modelos experimentales y proporcionan las bases para el desarrollo racional de nuevas terapias, aportando así un enfoque viable contra las complicaciones inflamatorias de la diabetesDiabetes is a chronic disease with a high social and clinical impact because of its high frequency and the consequences of its chronic complications. Among them, nephropathy and atherosclerosis are the leading cause of chronic kidney disease and morbidity and mortality in diabetic patients, although currently there are no therapeutic alternatives that effectively stop the occurrence of cardiovascular events or the progression to kidney failure. The inflammatory response associated to diabetic complications is mediated by numerous inflammatory proteins, whose gene expression is strictly regulated by several intracellular mechanisms. In this thesis we have studied the therapeutic potential of two intracellular pathways activated during the initiation and progression of diabetic nephropathy and atherosclerosis: NF-κB (nuclear factor-κB) and JAK/STAT (Janus kinase/Signal transducer and activator of transcription). In the first part, we analyzed the functional consequences of inhibiting specifically the IκB kinase (IKK)-dependent NF-κB pathway in an experimental model of diabetic nephropathy and atherosclerosis, the streptozotocin-induced diabetic apolipoprotein E deficient mouse (apoE-/-). For that, we used a cell permeable peptide that blocks the formation of the IKK complex. Treatment with the inhibitory peptide did not affect hyperglycemia and lipid profile in diabetic mice but improved their renal function by attenuating renal lesions (mesangial expansion, fibrosis and infiltrate), NF-κB activity and the expression of proinflammatory and profibrotic genes. Peptide administration also suppressed the NF-κB activity in diabetic mouse aortas, reduced the size, lipid content and inflammation (leucocytes and cytokines) in atherosclerotic lesions, while increasing plaque stabilization markers (collagen and vascular smooth muscle cells). In vitro, the peptide blocked the interaction between IKKγ and IKKα/β subunits, thus preventing nuclear translocation of NF-κB, target gene expression, and cell proliferation induced by hyperglycemic and inflammatory conditions. In the second part, we investigated the atheroprotective effect of JAK/STAT inhibition in apoE-/- mice by gene therapy with recombinant adenovirus encoding the suppressor of cytokine signaling 3 (SOCS3). SOCS3 transgene expression in aorta attenuated STAT1 and STAT3 activation and the expression of STAT-dependent genes (cytokines, chemokines, adhesion molecules and scavenger receptors), therefore preventing the development of atherosclerosis in early phases. Furthermore, SOCS3 gene delivery promoted a more stable plaque phenotype characterized by lower content of lipids, T cells and M1 macrophages and higher content of M2 macrophages and collagen. However, SOCS3 expression was not able to halt the progression of advanced lesions in atherosclerotic mice. Systemically, SOCS3 gene therapy inhibited the expression of Th1 and Th17 cytokines and reduced the activation state of circulating monocytes. In vitro, SOCS3 prevented cytokine-induced STAT activation, pro-inflammatory gene expression and cell proliferation in vascular cells and macrophages. In conclusion, our studies demonstrate the renoprotective and antiatherosclerotic effect of the selective inhibition of NF-κB and JAK/STAT pathways in experimental models and provide the basis for the rational development of new therapies, thus providing a feasible approach against the inflammatory complications of diabetes

Avances y limitaciones en la evaluación del aprendizaje a partir del proceso de convergencia. Visión docente y discente en los Grados de Educación Infantil y Primaria

Recent studies reveal punctual advances in some of the classicalmalfunctions of the university learning assessment. Its development is linked to the construction of a new assessment paradigm, based on the European Higher Education Area and featured by five main improvement challenges: Sensitive continuity of the evaluation, Strategic orientation to learning, Tasks and instruments for competency development and accreditation, Transparency and deep understanding of the assessment criteria and Student participation. This paper contrasts the perception of these challenges in a sample of students and teachers involved in educational degrees. The inquiry was carried out by means of a questionnaire, based on rubrics which detail five acceptance and fulfilment levels in relation to each challenge. The results disclose that the five of them are considered important and there is no divergence in the acceptance declared by both teachers and students. The differences found in the confidence interval have a small effect and refer to the degree of compliance particularly noticed in three of the challenges.Students describe a significantly inferior fulfilment when addressing thetransparency in assessment criteria and the sensitive continuity, connected to the lower use of the formative function of the assessment. Students recognise that the assessment does not only come at the end, but they do not think it is developed in a sensitive and integrated way, as teaching staff state. They also acknowledge having information about the assessment criteria, but they reject the carrying out of tasks focused on explaining the same. Both sectors of the sample recognise insufficient achievement in relation to the student participation, although the average from the students’ answers is significantly inferior. These results suggest that despite the advances observed on its form, the assessment still represents a mainly certifying tool.Estudios recientes revelan avances puntuales en algunas disfuncionalidades clásicas de la evaluación del aprendizaje universitario. Su desarrollo se vincula a la construcción de un nuevo paradigma evaluativo, ligado al Espacio Europeo de Educación Superior y caracterizado en cinco retos de mejora: Continuidad sensible de la evaluación, Orientación estratégica al aprendizaje, Tareas e instrumentos para el desarrollo y acreditación de competencias, Explicitación e interiorización de criterios y condiciones evaluativas y Participación discente. Este trabajo contrasta la visión de los mismos en una muestra de estudiantes y profesorado de los Grados de Educación Infantil y Primaria. La indagación se ha realizado por medio de un cuestionario, basado en rúbricas que detallan distintos grados de aceptación y cumplimiento en cada uno de los retos. Los resultados revelan que los cinco se juzgan relevantes, sin que se produzcan discrepancias en la aceptación declarada por docentes y discentes. Las diferencias detectadas dentro del intervalo de confianza tienen un efecto de tamaño pequeño y hacen referencia al grado de cumplimiento percibido en tres retos en particular. Al tratar la Explicitación de la evaluación y su Continuidad sensible, el alumnado describe un cumplimiento significativamente inferior, asociado a una menor explotación de la funcionalidad pedagógica de la evaluación. Los estudiantes reconocen que la evaluación no sólo se realiza al final, aunque tampoco creen que se desarrolle de manera sensible e integrada como afirma el profesorado. Asimismo, admiten disponer de información sobre las condiciones evaluativas, pero niegan la realización de tareas que contribuyan a su clarificación. Al valorar la Participación discente en la evaluación, ambos sectores de la muestra reconocen una consecución insuficiente, aunque los promedios de respuesta del alumnado son significativamente menores. Estos resultados sugieren que a pesar de los avances advertidos en la forma, en el fondo la evaluación sigue representando una herramienta principalmente acreditativa

The evaluative competence as a key factor in teaching quality: analysis on the pre-service teachers training

La competencia evaluativa docente destaca por su incidencia en la enseñanza, el aprendizaje y el desarrollo institucional. Su relevancia se hace especialmente manifiesta en la formación básica de maestras/os, donde el alumnado configura buena parte de las representaciones evaluativas que reproducirá al desempeñar el rol docente. El presente trabajo analiza la visión de una muestra de 704 estudiantes de los Grados de Educación Infantil y Primaria sobre la competencia evaluativa del profesorado. La recogida de información se ha realizado por medio de un cuestionario que capta el cumplimiento percibido en cinco factores vinculados a la evaluación: Diseño, Implementación, Control, Resultados y Revisión. El análisis estadístico ofrece una descripción general de la competencia evaluativa advertida por las/os estudiantes y la contrasta en función del grado de competencia docente que subjetivamente atribuyen al profesorado (poco competente, competenteo muy competente en su labor docente) competencia docente que subjetivamente atribuyen al profesorado (poco competente, competente o muy competente en su labor docente). Los resultados revelan percepciones significativamente diferentes sobre la práctica evaluativa desarrollada en los tres grupos docentes. La magnitud de tales diferencias es grande en el Diseño evaluativo y el Control técnico, y media para la Implementación, los Resultados y la Revisión. En todas ellas, los promedios que indican una mayor percepción de cumplimiento recaen sobre el profesorado considerado muy competente en la labor docente. Incluso en este caso, el alumnado percibe un amplio margen de mejora en el ejercicio de la competencia evaluativa. La Revisión de la evaluación y el feedback y la proacción a partir de los Resultados representan, a su juicio, los factores menos desarrollados.Teacher´s evaluative competence stands out for its impact in the teaching and learning process and in the institutional development. Its relevance is especially evident in the pre-service teachers training, where the students set much of the evaluative representations that will be played when accessing the teaching role. This study analyses the perception of a sample of 704 students of pre-school and primary education degrees on teacher evaluative competence. The collection of information was done by administering a questionnaire to gather the students’ perception on five evaluation factors: Design, Implementation, Monitoring, Results and Review. The analysis provides a general description of the evaluative competence noticed by the students and contrasts it according to the teaching competence that they subjectively attributed to the teaching staff (not very competent, competent or very competent). The students value significantly different evaluative practices among teachers considered not competent, competent and very competent in their teaching work. The magnitude of these differences is large in the case of evaluation Design and technical Monitoring, and medium for the Implementation, Results and Review. In all of them, the average indicating greater perception of compliance falls on teaching staff with best teaching competence rates. Even in this case, the students notice a wide range of improvement concerning the practise of the teaching evaluative competence. The Review of the assessment and the processes of feedback and pro-action based on the Results are, in their opinion, the least developed factors

Book review: Ralph R. Frerichs: Deadly river - Cholera and cover-up in post-earthquake

As most of PV systems, CPV systems are also affected by mismatching losses, particularly due to misalignment of optics and receivers. As a result, module level power electronics can help to increase their energy yield by making every CPV module deliver it maximum power at the output. Among the different alternatives, solutions based on DC power optimizers exhibit higher conversion efficiencies and lower costs than microinverters. However, while microinverters ensure optimal operation independently from the operating conditions, system design with DC power optimizers must be carefully examined to avoid potential underperformance. This paper describes not only the customized design and validation of a high-efficiency and economical DC power optimizer for HCPV systems, but also a comprehensive analysis of the whole system design to optimize its production under expected working conditions.Main design specifications (electrical parameters of module and number of modules per tracker), as well as supporting information for mismatching losses estimation, have been provided by ASSE within CPVMatch project. This work has received funding from the European Union’s Horizon 2020 research and innovation programme within the project CPVMatch under grant agreement No 640873

Incentive learning underlying cocaine relapse requires mGluR5 receptors located on dopamine D1 receptor-expressing neurons

Understanding the psychobiological basis of relapse remains a challenge in developing therapies for drug addiction. Relapse in cocaine addiction often occurs following exposure to environmental stimuli previously associated with drug taking. The metabotropic glutamate receptor, mGluR5, is potentially important in this respect; it plays a central role in several forms of striatal synaptic plasticity proposed to underpin associative learning and memory processes that enable drug-paired stimuli to acquire incentive motivational properties and trigger relapse. Using cell type-specific RNA interference, we have generated a novel mouse line with a selective knock-down of mGluR5 in dopamine D1 receptor-expressing neurons. Although mutant mice self-administer cocaine, we show that reinstatement of cocaine-seeking induced by a cocaine-paired stimulus is impaired. By examining different aspects of associative learning in the mutant mice, we identify deficits in specific incentive learning processes that enable a reward-paired stimulus to directly reinforce behavior and to become attractive, thus eliciting approach toward it. Our findings show that glutamate signaling through mGluR5 located on dopamine D1 receptor-expressing neurons is necessary for incentive learning processes that contribute to cue-induced reinstatement of cocaine-seeking and which may underpin relapse in drug addiction

Factors Associated With Peripheral Nerve Injury After Pelvic Laparoscopy: The Importance of Surgical Positioning

BACKGROUND: Nerve damage after abdominal and pelvic surgery is rare but potentially serious. The incidence of peripheral nerve injury is difficult to assess, and rates of between 0.02% and 21% have been cited in the literature. Signs and symptoms of this type of injury may appear immediately after surgery or a few days later. PURPOSE: This study was developed to assess the rate of peripheral nerve injury after pelvic laparoscopy and to identify associated risk factors. METHODS: A pilot prospective cohort study was conducted between March 2018 and April 2019 on 101 patients with a 1-month follow-up using two semistructured clinical interviews. We carried out a descriptive analysis followed by univariable and multivariable logistic regression analyses. RESULTS: Thirteen patients were found to have peripheral nerve injuries, representing a rate of 12.9%. Overall, 14 injuries (five severe and nine mild) were detected. One patient had two mild injuries. In this study, the risk of injury was found to increase 1.77-fold (OR = 1.77, 95% CI [1.13, 2.76], p = .007) for each hour the patient was in the Trendelenburg position. CONCLUSIONS/IMPLICATIONS FOR PRACTICE: The longer the patient is in the Trendelenburg position, the greater the risk of peripheral nerve damage. Patients aged 60 years or less also face a higher risk of nerve injury.This study was supported by a 600-euro grant from the Basque Foundation for Health Innovation and Research

Cocaine-induced behavioral sensitization is associated with changes in the expression of endocannabinoid and glutamatergic signaling systems in the mouse prefrontal cortex

Abstract Background: Endocannabinoids modulate the glutamatergic excitatory transmission by acting as retrograde messengers. A growing body of studies has reported that both signaling systems in the mesocorticolimbic neural circuitry are involved in the neurobiological mechanisms underlying drug addiction. Methods: We investigated whether the expression of both endocannabinoid and glutamatergic systems in the prefrontal cortex (PFC) were altered by an acute and/or repeated cocaine administration schedule that resulted in behavioral sensitization. We measured the protein and mRNA expression of the main endocannabinoid metabolic enzymes and the cannabinoid receptor type 1 (CB1). We also analyzed the mRNA expression of relevant components of the glutamatesignaling system, including glutamate-synthesizing enzymes, metabotropic receptors, and ionotropic receptors. Results: Although acute cocaine (10 mg/kg) produced no significant changes in the endocannabinoid-related proteins, repeated cocaine administration (20 mg/kg daily) induced a pronounced increase in the CB1 receptor expression. In addition, acute cocaine administration (10 mg/kg) in cocaine-sensitized mice (referred to as cocaine priming) induced a selective increase in the endocannabinoid-degrading enzymes fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase (MAGL). These protein changes were accompanied by an overall decrease in the ratios of endocannabinoid synthesis/degradation, especially the N-acyl phosphatidylethanolamine phospholipase D/FAAH and diacylglycerol lipase alpha/MAGL ratios.Regarding mRNA expression, while acute cocaine administration produced a decrease in CB1 receptors and N-acyl phosphatidylethanolamine phospholipase D, repeated cocaine treatment enhanced CB1 receptor expression. Cocaine sensitized mice that were administered priming injections of cocaine mainly displayed an increased FAAH expression. These endocannabinoid changes were associated with modifications in glutamatergic transmission-related genes. An overall decrease was observed in the mRNA expression of the glutamate-synthesizing gene kidney-type glutaminase (KGA), the metabotropic glutamate receptors (mGluR3 and GluR), and subunits of NMDA ionotropic receptors (NR1, NR2A, NR2B and NR2C) after acute cocaine administration, while mice repeatedly exposed to cocaine only displayed an increase in NR2C. However, in cocaine-sensitized mice primed with cocaine, this inhibition was reversed and a strong increase was detected in the mGluR5, NR2 subunits, and both GluR1 and GluR3. Conclusions: These findings indicate that cocaine sensitization is associated with an endocannabinoid downregulation and a hyperglutamatergic state in the PFC that, overall, contribute to an enhanced glutamatergic input into PFC-projecting areasThis work was supported by Ministerio de Ciencia e Innovación (PI13/02261 and SAF 2010–20521), Instituto de Salud Carlos III (ISCIII), Ministerio de Economía y Competitividad, Red de Trastornos Adictivos (RD12/0028/0001), Plan Nacional Sobre Drogas, Ministerio de Sanidad y Consumo (PNSD2013/049), Consejería de Economía, Innovación y Ciencia, Junta de Andalucía, UE/ERDF (CTS-433 and P-11-CVI-07637), Consejería de Salud, and Junta de Andalucía (PI0232/2008, PI0029/2008 and SAS111224). Dr Suárez is the recipient of a Miguel Servet research contract from ISCIII (CP12/03109). We thank Mariam Vázquez for English language assistance

Role of the satiety factor oleoylethanolamide in alcoholism

Oleoylethanolamide (OEA) is a satiety factor that controls motivational responses to dietary fat. Here we show that alcohol administration causes the release of OEA in rodents, which in turn reduces alcohol consumption by engaging peroxisome proliferator-activated receptor-alpha (PPAR-α). This effect appears to rely on peripheral signaling mechanisms as alcohol self-administration is unaltered by intracerebral PPAR-α agonist administration, and the lesion of sensory afferent fibers (by capsaicin) abrogates the effect of systemically administered OEA on alcohol intake. Additionally, OEA is shown to block cue-induced reinstatement of alcohol-seeking behavior (an animal model of relapse) and reduce the severity of somatic withdrawal symptoms in alcohol-dependent animals. Collectively, these findings demonstrate a homeostatic role for OEA signaling in the behavioral effects of alcohol exposure and highlight OEA as a novel therapeutic target for alcohol use disorders and alcoholism

Cannabinoid CB1 receptor antagonism markedly increases dopamine receptor-mediated stereotypies

The contribution of the endocannabinoid system to dopamine-mediated disorganized behavior in schizophrenia is discussed. We used a model of concurrent stimulation of dopamine D1 and D2 receptors to evaluate the role of this system in dopamine-mediated stereotypies measured in a hole-board test. Pretreatment with the cannabinoid CB1 receptor antagonist N-(piperidin-1-yl)-5-(4-chlorophenyl)-1-(2,4-dichloro-phenyl)-4-methyl-1H-pyrazole-3-carboxamide (SR141716A; 1 mg/kg) potentiated stereotyped behavior induced by coadministration of the dopamine D1 receptor agonist SKF 38393 (0.05, 0.1 and 1 mg/kg) and the dopamine D2 receptor agonist quinpirole (0.25 mg/kg). Thus, the endocannabinoid system acts as a brake for abnormal behavior associated with dopaminergic overactivation. © 2007 Elsevier B.V. All rights reserved.Peer Reviewe