Estudio de las mucinas ancladas a membrana como diana farmacológica en la fibrosis pulmonar

Antecedentes: La fibrosis pulmonar idiopática (FPI) es una enfermedad pulmonar intersticial fibrosante y crónica de etiología desconocida e incurable englobada dentro de las enfermedades raras. Varias mucinas han demostrados estar implicadas en esta enfermedad, sin embargo, no hay evidencia hasta la fecha sobre el papel de la mucina anclada a membrana MUC4 en el desarrollo de FPI. Objetivo: Evaluar la sobreexpresión y localización de MUC4 en el tejido pulmonar de pacientes con FPI asi como caracterizar su posible implicación en la activación y transformaciones celulares que participan en la FPI (proliferación de fibroblastos, senescencia, transición epitelio mesénquima y transición fibroblasto mesénquima), así como los aspectos funcionales característicos como base preclínica y traslacional de futuros tratamientos. Métodos y resultados: la expresión de MUC4 aumentó en el tejido pulmonar de pacientes con FPI (n = 22) en comparación con el de los sujetos sanos (n = 21) y se localizó en arterias pulmonares, células epiteliales bronquiales basales, fibroblastos de la submucosa y células alveolares hiperplásicas de tipo II. TGF-B1 promovió las transformaciones mesenquimatosas / miofibroblásticas aumentando la expresión de colágeno tipo I, Snail, Slug y actina de músculo liso alfa, en células alveolares tipo II (A549) y fibroblastos de pulmón humano (MRC5), y disminuyó el marcador de células epiteliales E-cadherina en las células A549. La disminución de la expresión de MUC4 usando siRNA-MUC4 inhibió las transformaciones tanto de epitelio a mesénquima (TEM) con de fibroblasto a miofibroblasto /características de esta enfermedad de forma estadísticamente significativa,así como la senescencia celular y la proliferación de fibroblastos inducida por TGFB-1. La sobreexpresión de MUC4 en las células HEK293 transfectadas con un sistema de expresión génica de MUC4 inducible con doxiciclina, aumentó los efectos de TGFB-1 sobre las transformaciones mesenquimales / miofibroblastos y la senescencia celular. La sobreexpresión de MUC4 y el silenciamiento del gen siRNA-MUC4 aumentaron o disminuyeron respectivamente la fosforilación de TGFB-RI y SMAD3 contribuyendo a la activación del elemento de unión de smad a nivel del núcleo, que regula las respuestas transcripcionales que inducen los diferentes procesos implicados en la FPI. El análisis de inmunoprecipitación y la inmunofluorescencia cofocal mostraron la formación de un complejo proteico entre MUC4B / p-TGFB-RI y tambien con SMAD3 fosforilado en la membrana celular después de la estimulación con TGFB-1 y en el tejido pulmonar de pacientes con FPI. La fibrosis pulmonar inducida por bleomicina y la mortalidad secundaria se redujo en ratones transfectados transitoriamente con siRNA-MUC4. También se observó una menor afectación de la función pulmonar medida por pletismografía de cuerpo entero en los ratones transfectados transitoriamente con siRNA-MUC4 y una menor respuesta inflamatoria que en los ratones no silenciados y tratados con bleomicina. La administración de bleomicina en los ratones nativos produjo un aumento de la expresión génica del ARN de marcadores fibróticos reconocidos como colágeno tipo I, TGFB1, CRGF, IL-13, MUC5B y marcadores fibróticos oxidativos NOX4 y NRF-2, siendo esta elevación significativamente menor en los ratones con silenciamiento de MUC4. De la misma manera que el tejido pulmonar humano estudiamos la colocalización de MUC4 con TGFB-R1 fosforilado y SMAD3 fosforilado mediante microscopía cofocal. En los ratones control con fibrosis inducida por bleomicina observamos una distribución citoplasmática y cofocal de ambos complejos proteicos siendo este fenómeno prácticamente indetectable en los ratones con MU$ silenciado y en los ratones control no tratados con bleomicina. Conclusiones: la expresión de MUC4 aumenta en la FPI y promueve procesos fibróticos en colaboración con la vía canónica de TGFB-1 que podría ser un objetivo farmacológico atractivo para el tratamiento de la FPI humana

Diseño e implantación de una red inalámbrica unificada en el Colegio Nuestra Señora de Fátima de Valencia

El objetivo del presente trabajo es mostrar como se ha diseñado, implantado y configurado, en el colegio Nuestra Señora de Fátima de Valencia, una red de comunicaciones inalámbrica unificada. Esta red permite dar cobertura a las seis plantas del edificio con zonas destinadas a fines académicos y con una velocidad adecuada a la demanda actual. Además, se puede gestionar de forma centralizada y casi automática, de ahí su nombre “unificada”. El número de puntos de acceso inalámbricos necesarios se determina a partir del número de posibles usuarios potenciales y de las áreas a cubrir en las diferentes plantas del edificio. Un estudio detallado de la propagación radioeléctrica de la red, determina el número de los puntos de acceso necesarios definitivo y su óptima ubicación para cubrir todas las zonas requeridas. Una vez emplazados los puntos de acceso, se diseña la arquitectura de la red cableada, que junto con todo el equipamiento necesario, permite la conexión a Internet y a la LAN existente. Finalmente se configuran los dispositivos (controladora, switch y servidor RADIUS) encargados de la gestión, la autenticación, la segmentación, la seguridad y del buen funcionamiento de la redThe aim of this work is to show as designed, installed and configured in the school Nuestra Señora de Fatima of Valencia, a unified wireless communications network. This network allows to cover all six floors of the building with areas intended for academic purposes and with adequate speed to the current demand. In addition, it can centrally manage and almost automatically, hence its name "unified". The number of wireless access points required is determined from the number of potential users and possible areas to be covered in the different floors. A detailed study of radio propagation network, determines the definitive number of access points needed and the optimal location to cover all required areas. Once deployed access points, the architecture of the wired network, with all the necessary equipment, allows connection to the Internet and the existing LAN is designed. Finally the devices (controller, switch and RADIUS server) responsible for the management, the authentication, the segmentation, the security and the proper functioning of the network are configuredMurillo Safont, JM. (2015). Diseño e implantación de una red inalámbrica unificada en el Colegio Nuestra Señora de Fátima de Valencia. Universitat Politècnica de València. http://hdl.handle.net/10251/57385TFG

On the Birman invariants of Heegaard splittings

J. Birman had observed that the homological information about a given Heegaard splitting of genus g is contained in a double coset in the group of symplectic 2g×2g integer matrices with respect to a suitable subgroup, and found a determinant invariant of this double coset. We obtain complete invariants of these double cosets by characterizing it in terms of the linking form of the manifold lifted to a handlebody of the Heegaard splitting and then finding complete invariants of this lifted form

Effect of the Purification Treatment on the Valorization of Natural Cellulosic Residues as Fillers in PHB-Based Composites for Short Shelf Life Applications

This is a pre-print of an article published in Waste and Biomass Valorization. The final authenticated version is available online at: https://doi.org/10.1007/s12649-020-01192-1In this work the effect of a combined NaOH + peracetic acid (PAA) purification treatment on the valorization of almond shell (AS) and rice husk (RH) lignocellulosic residues as fillers in PHB-based composites for short shelf life applications has been studied. The efficiency of the treatment at removing the non-cellulosic components of the fibers has been evaluated by SEM, FTIR, WAXS and TGA taking a commercial cellulose as reference. The influence of the untreated and treated fibers on the morphology, thermal, crystallization, tensile properties, fracture toughness and dynamo mechanical behavior of the PHB/fiber composites has been studied. The treatment has demonstrated its ability at removing the lignin, hemicelluloses and waxes allowing the obtention of fibers with relative crystallinity, thermal stability and composition similar to the commercial cellulose. The different agro-food based lignocellulosic residues used resulted in two suitable reinforcing fillers for a PHB matrix. Hence, composites prepared with the treated fibers presented better thermal and mechanical performance than those prepared with the untreated ones. Therefore, the so-obtained purified residue fibers are comparable to a pure cellulose as a filler for PHB composites

Poly(3-Hydroxybutyrate-co-3-Hydroxyvalerate)/ Purifi ed Cellulose Fiber Composites by Melt Blending: Characterization and Degradation in Composting Conditions

Novel biodegradable composites based on poly(3-hydroxybutirate-co-3-hydroxyvalerate) (PHBV) and different contents of purifi ed alpha-cellulose fi bers (3, 10, 25 and 45%) were prepared by melt blending and characterized. The composites were characterized by scanning electron microscopy (SEM), wide-angle X-ray scattering (WAXS) experiments, thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), dynamic mechanic analysis (DMA) and Shore D hardness measurements. Disintegrability under composting conditions was studied according to the ISO 20200 standard. Morphological results showed that high dispersion of the fi bers was achieved during mixing. Good adhesion on the fi ber-matrix interface was also detected by SEM. The addition of low and medium cellulose contents did not result in lower thermal resistance with respect to the neat PHBV. A reinforcing effect of the cellulose fi bers was detected in all samples, this effect being more pronounced at high temperatures. The composting results show that the addition of the fi bers did not affect the disintegrability of the PHBV, and thus compostable “green” low-cost PHBV/cellulose composites can be obtained

Study of the Compatibilization Effect of Different Reactive Agents in PHB/Natural Fiber-Based Composites

Fiber–matrix interfacial adhesion is one of the key factors governing the final properties of natural fiber-based polymer composites. In this work, four extrusion reactive agents were tested as potential compatibilizers in polyhydroxylbutyrate (PHB)/cellulose composites: dicumyl peroxide (DCP), hexamethylene diisocyanate (HMDI), resorcinol diglycidyl ether (RDGE), and triglycidyl isocyanurate (TGIC). The influence of the fibers and the different reactive agents on the mechanical properties, physical aging, and crystallization behavior were assessed. To evaluate the compatibilization effectiveness of each reactive agent, highly purified commercial cellulose fibers (TC90) were used as reference filler. Then, the influence of fiber purity on the compatibilization effect of the reactive agent HMDI was evaluated using untreated (U_RH) and chemically purified (T_RH) rice husk fibers, comparing the results with the ones using TC90 fibers. The results show that reactive agents interact with the polymer matrix at different levels, but all compositions showed a drastic embrittlement due to the aging of PHB. No clear compatibilization effect was found using DCP, RDGE, or TGIC reactive agents. On the other hand, the fiber–polymer interfacial adhesion was enhanced with HMDI. The purity of the fiber played an important role in the effectiveness of HMDI as a compatibilizer, since composites with highly purified fibers showed the greatest improvements in tensile strength and the most favorable morphology. None of the reactive agents negatively affected the compostability of PHB. Finally, thermoformed trays with good mold reproducibility were successfully obtained for PHB/T_RH/HMDI composition

A randomized phase II study of capecitabine-based chemoradiation with or without bevacizumab in resectable locally advanced rectal cancer: clinical and biological features

Background: perioperatory chemoradiotherapy (CRT) improves local control and survival in patients with locally advanced rectal cancer (LARC). The objective of the current study was to evaluate the addition of bevacizumab (BEV) to preoperative capecitabine (CAP)-based CRT in LARC, and to explore biomarkers for downstaging. Methods: patients (pts) were randomized to receive 5 weeks of radiotherapy 45 Gy/25 fractions with concurrent CAP 825 mg/m2 twice daily 5 days per week and BEV 5 mg/kg once every 2 weeks (3 doses) (arm A), or the same schedule without BEV (arm B). The primary end point was pathologic complete response (ypCR: ypT0N0). Results: ninety pts were included in arm A (44) or arm B (46). Grade 3-4 treatment-related toxicity rates were 16% and 13%, respectively. All patients but one (arm A) proceeded to surgery. The ypCR rate was 16% in arm A and 11% in arm B (p =0.54). Fifty-nine percent vs 39% of pts achieved T-downstaging (arm A vs arm B; p =0.04). Serial samples for biomarker analyses were obtained for 50 out of 90 randomized pts (arm A/B: 22/28). Plasma angiopoietin-2 (Ang-2) levels decreased in arm A and increased in arm B (p <0.05 at all time points). Decrease in Ang-2 levels from baseline to day 57 was significantly associated with tumor downstaging (p =0.02). Conclusions: the addition of BEV to CAP-based preoperative CRT has shown to be feasible in LARC. The association between decreasing Ang-2 levels and tumor downstaging should be further validated in customized studies

Phase II randomized trial of capecitabine with bevacizumab and external beam radiation therapy as preoperative treatment for patients with resectable locally advanced rectal adenocarcinoma: long term results

Background: Preoperative chemoradiotherapy with capecitabine is considered as a standard of care for locally advanced rectal cancer. The “Tratamiento de Tumores Digestivos” group (TTD) previously reported in a randomized Ph II study that the addition of Bevacizumab to capecitabine-RT conferred no differences in the pre-defined efficacy endpoint (pathological complete response). We present the follow-up results of progression-free survival, distant relapse-free survival, and overall survival data at 3 and 5 years. Methods: Patients (pts) were randomized to receive 5 weeks of radiotherapy (45 Gy/25 fractions) with concurrent Capecitabine 825 mg/m2 twice daily, 5 days per week with (arm A) or without (arm b) bevacizumab (5 mg/kg once every 2 weeks). Results: In our study, the addition of bevacizumab to capecitabine and radiotherapy in the neoadjuvant setting shows no differences in pathological complete response (15.9% vs 10.9%), distant relapse-free survival (81.0 vs 80.4 and 76.2% vs 78.2% at 3 and 5 years respectively), disease-free survival (75% vs 71.7 and 68.1% vs 69.57% at 3 and 5 years respectively) nor overall survival at 5-years of follow-up (81.8% vs 86.9%). Conclusions: the addition of bevacizumab to capecitabine plus radiotherapy does not confer statistically significant advantages neither in distant relapse-free survival nor in disease-free survival nor in Overall Survival in the short or long term

Biocomposites of different lignocellulosic wastes for sustainable food packaging applications

The suitability of three local lignocellulosic wastes i.e. almond shell (AS), rice husk (RH) and seagrass (SG) as fillers in PHB/Fiber composites applications has been studied. PHB/Fiber composites with 10 phr and 20 phr fiber content were prepared by melt blending. The influence of the fiber type (size, morphology and origin) and content on the morphological, mechanical and thermal properties of the as obtained composites has been assessed. To evaluate the potential use in food packaging applications, the barrier performance to water, thermoforming ability and disintegration in controlled composting conditions of the composites were also studied. All the fibers have demonstrated to be apt for their use as fillers in PHB/Fiber composites, showing a reinforcing effect without affecting the crystallinity and the disintegration rate of PHB. The thermal stability and the water barrier performance of the composites were reduced by the presence of the fibers. Nevertheless, the addition of AS resulted in the best balance of properties, in terms of permeability and mechanical properties, finding an enhancement of the thermoforming ability of PHB when 10 phr of AS was added