Generation and characterization of a new conditional mouse model of Hutchinson-Gilford Progeria Syndrome to assess disease progression upon progerin suppression and lamin A restoration

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Bioquímica. Fecha de lectura: 24-01-2020Esta tesis tiene embargado el acceso al texto completo hasta el 24-07-2021El Síndrome de Progeria de Hutchinson-Gilford (HGPS, por sus siglas en inglés) es un trastorno genético fatal extremadamente raro caracterizado por envejecimiento acelerado y muerte prematura a una edad promedio de 14,6 años. El HGPS "clásico" está causado por una mutación de novo c.1824 C>T en el exón 11 del gen LMNA, que codifica las laminas A y C. Se trata de una mutación que, a pesar de ser sinónima, promueve la expresión de progerina, una forma anómala de prelamina A que no puede madurar correctamente, permaneciendo farnesilada y unida a la membrana nuclear interna, afectando muchos procesos celulares de manera dominante-negativa. Los pacientes con HGPS tienen un aspecto normal al nacer, pero desarrollan síntomas de la enfermedad típicamente durante el primer y segundo año de vida. Teniendo en cuenta que no existe una cura definitiva para HGPS y que los pacientes son diagnosticados cuando los síntomas están ya presentes, es crítico determinar si el daño causado por la progerina es reversible o si la progresión de HGPS puede disminuir o detenerse mediante la supresión de la expresión de esta proteína. También es necesario investigar la contribución relativa de factores sistémicos y específicos de tejido al desarrollo de HGPS, para evaluar la efectividad de posibles terapias futuras diseñadas para suprimir la expresión de progerina en tejidos específicos, lo que sería más sencillo que la supresión de progerina en todo el cuerpo. Para abordar estas preguntas, en esta Tesis Doctoral utilizamos el sistema CRISPR-Cas9 para generar ratones LmnaHGPSrev, el primer modelo de progeria “reversible” con expresión ubicua de progerina que puede suprimirse de manera controlada espacio-temporalmente tras la activación de la recombinasa Cre, que además permite el restablecimiento de la expresión de la lamina A. Hemos demostrado que los ratones LmnaHGPSrev/HGPSrev recapitulan las características principales de HGPS en humanos, incluyendo alteraciones en el desarrollo, problemas cardiovasculares y muerte prematura. Además, mostramos que el tratamiento de estos ratones progeroides mediante la activación de la Cre con tamoxifeno en una etapa avanzada de la enfermedad aumenta su supervivencia. Finalmente, la terapia génica con virus adeno-asociados que sobreexpresan Cre mejora el crecimiento postnatal de ratones progéricos adultos con síntomas iniciales de HGPS. Estos y futuros estudios con ratones LmnaHGPSrev/HGPSrev más allá del alcance de la presente Tesis Doctoral, arrojarán información importante sobre los mecanismos celulares y moleculares de HGPS y facilitarán el camino para desarrollar terapias más eficientes.Hutchinson-Gilford progeria syndrome (HGPS) is a rare fatal genetic disorder characterized by accelerated aging and premature death at an average age of 14.6 years. “Classical” HGPS is caused by a heterozygous de novo c.1824 C>T dominant synonymous point mutation in the LMNA gene, which encodes for lamin A and C. This mutation promotes the expression of a mutant protein called progerin, an aberrant form of prelamin A that cannot undergo complete maturation. Progerin remains permanently farnesylated and firmly anchored to the inner nuclear membrane, affecting many cellular processes in a dominant-negative manner. HGPS patients appear normal at birth but develop symptoms of the disease typically during the first and second year of life. Taking into account that there is still no definitive cure for HGPS and that patients are diagnosed when symptoms are already present, it is critically important to ascertain whether the damage caused by progerin expression is reversible or if disease progression can be slowed down or halted upon progerin suppression. It is also necessary to investigate the relative contribution of systemic and tissue-specific factors to the development of HGPS to assess the effectiveness of potential future therapies designed to suppress progerin expression in specific tissues, which, if proven effective, would likely be less challenging than whole-body progerin suppression. In order to address these questions, in this Doctoral Thesis we use the CRISPR-Cas9 system to generate LmnaHGPSrev mice, the first “reversible” mouse model of progeria which expresses progerin ubiquitously and allows a controlled spatio-temporal suppression of progerin expression with concomitant restoration of lamin A expression upon activation of the Cre recombinase. We demonstrate that LmnaHGPSrev/HGPSrev mice recapitulate the main features of human HGPS, including failure to thrive, cardiovascular alterations and premature death. Moreover, we show that Cre activation upon tamoxifen treatment starting at an advanced stage of the disease in LmnaHGPSrev/HGPSrev Ubc-CreERTtg/+ progeroid mice prolongs their life span. Finally, we show that gene therapy with adeno-associated virus overexpressing Cre ameliorates postnatal growth of adult LmnaHGPSrev/HGPSrev mice with initial symptoms of HGPS at the initiation of the therapy. These studies with the LmnaHGPSrev mouse model, and future studies beyond the scope of the present Doctoral Thesis, will shed significant light on the cellular and molecular mechanisms of HGPS and pave the way to developing more efficient therapies

Urban domestic architecture of the city of Arucci (Aroche, Huelva): an approach based on the Domus de Peristilo

La vivienda urbana puede aportar información de una enorme relevancia para la reflexión sobre el pasado romano al dar cabida al planteamiento y resolución de cuestiones de muy diversa naturaleza sobre la sociedad. El presente trabajo contempla el estudio arquitectónico y funcional de la Domus de Peristilo de la ciudad hispanorromana de Arucci (Aroche, Huelva), con el objetivo de presentar una panorámica general sobre la vivienda y la arquitectura doméstica de la ciudad, pero también sobre el marco urbano, económico, político, social y cultural en el que se inserta. Con ello, trataremos de conciliar la situación dispar que caracteriza a este tipo de estudios, tradicionalmente relegados a un segundo plano en favor del análisis de las grandes construcciones públicas, a la par que subrayaremos la necesidad de verlos incrementados en un futuro a corto y medio plazo.Urban housing can provide information of enormous relevance for reflection on the Roman past by allowing for the approach and resolution of questions of a very diverse nature about society. This paper will contemplate the architectural and functional study of the Domus de Peristilo of the hispanorroman city of Arucci (Aroche, Huelva), in order to present a general overview of housing and the domestic architecture of the city, but also of the urbanism, economic, political, social and cultural framework in which it is inserted. In doing so, we will attempt to reconcile the disparate situation that characterizes this type of studies, traditionally relegated to a secondary plane of importance in favor of the analysis of monumental public buildings, while we will stressing the need to increase them in a short and medium-term future

Reingeniería de reactor de lecho fluidizado

El gasificador de biomasa transforma los desechos orgánicos (generalmente de procedencia agrícola) en un gas limpio y de alta calidad, con unas propiedades específicas para emplearse en motores de combustión interna o en calderas. Puede llegar a reemplazar a los combustibles fósiles y proveer de energía eléctrica y calor. El objetivo de este proyecto es el diseño de un gasificador de laboratorio, con base en un reactor ya existente. Este reactor fue construído sin planos previos, unos años atrás. Con este proyecto se pretende rediseñarlo atendiendo a las modificaciones y mejoras que supone el paso de los años. La metodología que se va a llevar a cabo va a ser el cálculo preliminar y dimensionamiento preliminar del reactor ya existente com su posterior confección de los diseños constructivos

State-of-the-art polymeric nanoparticles as promising therapeutic tools against human bacterial infections

Infectious diseases kill over 17 million people a year, among which bacterial infections stand out. From all the bacterial infections, tuberculosis, diarrhoea, meningitis, pneumonia, sexual transmission diseases and nosocomial infections are the most severe bacterial infections, which affect millions of people worldwide. Moreover, the indiscriminate use of antibiotic drugs in the last decades has triggered an increasing multiple resistance towards these drugs, which represent a serious global socioeconomic and public health risk. It is estimated that 33,000 and 35,000 people die yearly in Europe and the United States, respectively, as a direct result of antimicrobial resistance. For all these reasons, there is an emerging need to find novel alternatives to overcome these issues and reduced the morbidity and mortality associated to bacterial infectious diseases. In that sense, nanotechnological approaches, especially smart polymeric nanoparticles, has wrought a revolution in this field, providing an innovative therapeutic alternative able to improve the limitations encountered in available treatments and capable to be effective by theirselves. In this review, we examine the current status of most dangerous human infections, together with an in-depth discussion of the role of nanomedicine to overcome the current disadvantages, and specifically the most recent and innovative studies involving polymeric nanoparticles against most common bacterial infections of the human body.Authors acknowledge the support of the Spanish Ministry of Economy and Competitiveness (SAF2017-84283-R and RTI2018-098641-B-I00), Biomedical Research Networking Centre in Neurodegenerative Diseases (CIBERNED, CB06/05/0024), Scientifc Project Marató TV3 (ref 201829-10) and European Regional Development Founds. Authors also acknowledge the Portuguese Science and Technology Foundation (FCT) for the strategic fund (UIDB/04469/2020).info:eu-repo/semantics/publishedVersio

Epilepsy in Neurodegenerative Diseases: Related Drugs and Molecular Pathways

Alzheimer’s disease; Huntington’s disease; Parkinson’s diseaseEnfermedad de Alzheimer; Enfermedad de Huntington; Enfermedad de ParkinsonMalaltia d'Alzheimer; Malaltia de Huntington; Malaltia de ParkinsonEpilepsy is a chronic disease of the central nervous system characterized by an electrical imbalance in neurons. It is the second most prevalent neurological disease, with 50 million people affected around the world, and 30% of all epilepsies do not respond to available treatments. Currently, the main hypothesis about the molecular processes that trigger epileptic seizures and promote the neurotoxic effects that lead to cell death focuses on the exacerbation of the glutamate pathway and the massive influx of Ca2+ into neurons by different factors. However, other mechanisms have been proposed, and most of them have also been described in other neurodegenerative diseases, such as Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, or multiple sclerosis. Interestingly, and mainly because of these common molecular links and the lack of effective treatments for these diseases, some antiseizure drugs have been investigated to evaluate their therapeutic potential in these pathologies. Therefore, in this review, we thoroughly investigate the common molecular pathways between epilepsy and the major neurodegenerative diseases, examine the incidence of epilepsy in these populations, and explore the use of current and innovative antiseizure drugs in the treatment of refractory epilepsy and other neurodegenerative diseases.A.C. acknowledges the support of the Spanish Ministry of Science, Innovation and Universities under the grant Juan de la Cierva (FJC2018-036012-I). Authors acknowledge the support of the Instituto de Salud Carlos III (ISCIII) Acción Estratégica en Salud, integrated into the Spanish National R+D+I Plan and financed by ISCIII Subdirección General de Evaluación and the Fondo Europeo de Desarrollo Regional (FEDER “Una manera de hacer Europa”) grant PI17/01474 awarded to M.B. Boada, grant PI19/00335 awarded to M.M. and the European Social Fund (ESF “Investing in your future”) for the Sara Borrell Contract (CD19/00232) to SA-L; M.E. acknowledges the support of the Spanish Ministry of Economy and Competitiveness under the project SAF2017-84283-R, and CIBERNED under project CB06/05/0024. E.B.S. acknowledges the support of the Portuguese Science and Technology Foundation (FCT) for the strategic fund (UIDB/04469/2020). A.R. acknowledges the support of CIBERNED (Instituto de Salud Carlos III (ISCIII)), the EU/EFPIA Innovative Medicines Initiative Joint Undertaking, ADAPTED Grant Nº 115975, from EXIT project, EU Euronanomed3 Program JCT2017 Grant Nº AC17/00100, from PREADAPT project. Joint Program for Neurodegenerative Diseases (JPND) Grant No. AC19/00097, and from grants PI13/02434, PI16/01861 BA19/00020, and PI19/01301. Acción Estratégica en Salud, integrated in the Spanish National RCDCI Plan and financed by Instituto de Salud Carlos III (ISCIII)- Subdirección General de Evaluación and the Fondo Europeo de Desarrollo Regional (FEDER—“Una manera de Hacer Europa”), by Fundación bancaria “La Caixa” and Grífols SA (GR@ACE project)

Surface functionalization of PLGA nanoparticles to increase transport across the BBB for Alzheimers disease

Alzheimers disease (AD) is a chronic neurodegenerative disorder that accounts for about 60% of all diagnosed cases of dementia worldwide. Although there are currently several drugs marketed for its treatment, none are capable of slowing down or stopping the progression of AD. The role of the blood-brain barrier (BBB) plays a key role in the design of a successful treatment for this neurodegenerative disease. Nanosized particles have been proposed as suitable drug delivery systems to overcome BBB with the purpose of increasing bioavailability of drugs in the brain. Biodegradable poly (lactic-co-glycolic acid) nanoparticles (PLGA-NPs) have been particularly regarded as promising drug delivery systems as they can be surface-tailored with functionalized molecules for site-specific targeting. In this review, a thorough discussion about the most recent functionalization strategies based on PLGA-NPs for AD and their mechanisms of action is provided, together with a description of AD pathogenesis and the role of the BBB in brain targeting.A.C. [Amanda Cano] acknowledges the support of the Spanish Ministry of Science, Innovation and Universities under the grant Juan de la Cierva (FJC2018-036012-I). Authors acknowledge the support of the Spanish Ministry of Economy and Competitiveness under the project SAF2017-84283-R; Biomedical Research Networking Centre in Neurodegenerative Diseases (CIBERNED, CB06/05/0024) and Portuguese Science and Technology Foundation (FCT) for the strategic fund (UIDB/04469/2020).info:eu-repo/semantics/publishedVersio

Pyrites in a salt marsh-ria system: quantification, morphology, and mobilization

Galician Rias are among the most productive ecosystems in the world. Consequently, the soils of their salt marshes and sediments of the intertidal flats show high organic matter contents, reactive Fe, and sulfate, which promote pyrite synthesis and accumulation, using sulfate for organic matter decomposition. This work studies the morphological variability and concentration of pyrites (individual crystals and framboids) in different geochemical environments found in the Ria de Ortigueira (salt marsh soils and bottom sediments in the inner, middle, and outer section), addressing their dynamics in the marsh-ria system in relation to the hydrodynamic characteristics defined by tides and river discharges. Framboidal pyrites were the dominant morphology in marsh soils and sediments in the middle and inner sections of the Ria, while isolated crystals dominated its outer section. The results showed that lower marsh soils (colonized by Spartina) are the most favorable environment for pyrite synthesis, showing high pyritic Fe concentrations and high degrees of pyritization, largely exceeding the values observed in sediments from Galician Rias and from most sedimentary environments worldwide. However, the amount of framboidal pyrites present in the lower marsh (SPE: 4–5 × 104 framboids) was clearly lower than in bottom sediments of the inner and middle part of the Ria de Ortigueira (∼2–7 × 106 framboids), mainly due to the fact that pyrites were found to form large framboids in lower salt marsh soils. Thus, the amount of framboidal pyrites does not seem to be a good indicator of redox conditions in modern marine sediments. Pyrite crystals found in the sediments of the Ria showed poorly defined vertices and facets, indicating their degradation and suggesting that a significant amount of the pyrites found in the middle and inner sections derive from marsh collapse. Finally, the output of framboidal pyrites towards the outer Ria de Ortigueira reflects the low intensity of residual flows in this Ria. Therefore, the pyrites observed in the outer section consisted only of isolated crystals, presumably formed in situ under low sulfate-reducing activity conditionsS

Estudio de pre factibilidad para la instalación de un Laboratorio de calibración de equipos de medición directa de riesgos higiénicos industriales en la Facultad de Tecnología de la Industria de la Universidad Nacional de Ingeniería

Presenta un estudio de instalación del laboratorio de calibración de equipos de medición directa de factores de riesgo higiénico industriales será el primero a nivel nacional en brindar este servicio, no contando con estudios previos que ofrezcan datos históricos sobre variables como precio, demanda u oferta, el laboratorio estará ubicado en el Recinto Universitario Pedro Arauz Palacios, en el espacio ocupado actualmente por el Laboratorio de Seguridad Industrial y estará diseñado para ofrecer 641 calibraciones para sonómetros al año (área de acústica) y 770 calibraciones para luxómetros al año (área de óptica). El proceso de calibración de sonómetros estará determinada por la Norma IEC 61672 partes 1, 2,y 3 en el caso de la calibración de luxómetros no existe una norma que defina un método de calibración por tanto se utilizará el método por comparación que es utilizado por el laboratorio mexicano MEYLAB, el laboratorio estará bajo la personería jurídica de la Universidad Nacional de Ingeniería y debe estar acreditado por la Oficina Nacional de Acreditaciones

Complementary pharmacological and toxicological characterization data on the pharmacological profile of N-(2,6-dichlorophenyl)-2-(4-methyl-1-piperidinyl) acetamide

This text presents complementary data corresponding to pharmacological and toxicological characterization of N-(2,6-dichlorophenyl)-2-(4-methyl-1-piperidinyl)acetamide (LIA) compound. These data support our research article entitled “Pharmacological profile of N-(2,6-dichlorophenyl)-2-(4-methyl-1-piperidinyl)acetamide, a novel analog of lidocaine” Déciga-Campos M., Navarrete-Vázquez G., López-Muñoz F.J., Librowski T., Sánchez-Recillas A., Yañez-Pérez V., Ortiz-Andrade R. (2016) [1]. Toxicity was predicted through the ACD/ToxSuite software and evaluated in vivo using brine shrimp larvae (Artemia salina L.) and mice. Also, we used the micronucleus assay to determine genotoxicity. We used the platform admetSAR to predict absorption properties of LIA and lidocaine. Keywords: N-(2,6-dichlorophenyl)-2-(4-methyl-1-piperidinyl)acetamide, Toxicity, Lidocain