FungalBraid 2.0: expanding the synthetic biology toolbox for the biotechnological exploitation of filamentous fungi

[EN] Fungal synthetic biology is a rapidly expanding field that aims to optimize the biotechnological exploitation of fungi through the generation of standard, readyto-use genetic elements, and universal syntax and rules for contributory use by the fungal research community. Recently, an increasing number of synthetic biology toolkits have been developed and applied to filamentous fungi, which highlights the relevance of these organisms in the biotechnology field. The FungalBraid (FB) modular cloning platform enables interchangeability of DNA parts with the GoldenBraid (GB) platform, which is designed for plants, and other systems that are compatible with the standard Golden Gate cloning and syntax, and uses binary pCAMBIA-derived vectors to allow Agrobacterium tumefaciensmediated transformation of a wide range of fungal species. In this study, we have expanded the original FB catalog by adding 27 new DNA parts that were functionally validated in vivo. Among these are the resistance selection markers for the antibiotics phleomycin and terbinafine, as well as the uridine-auxotrophic marker pyr4. We also used a normalized luciferase reporter system to validate several promoters, such as PpkiA,P7760,Pef1¿, and PafpB constitutive promoters, and PglaA,PamyB, and PxlnA inducible promoters. Additionally, the recently developed dCas9-regulated GB_SynP synthetic promoter collection for orthogonal CRISPR activation (CRISPRa) in plants has been adapted in fungi through the FB system. In general, the expansion of the FB catalog is of great interest to the scientific community since it increases the number of possible modular and interchangeable DNA assemblies, exponentially increasing the possibilities of studying, developing, and exploiting filamentous fungi.This work was supported by PROMETEO/2018/066 from "Conselleria d'Educacio" (Generalitat Valenciana, Comunitat Valenciana, Spain), grant PID2021-125858OB-100, and the Severo Ochoa Excellence Program CEX 2021-001189-S funded by MCIN/AEI/10.13039/501100011033 and by "ERDF A way of making Europe." EM-G was the recipient of a predoctoral grant FPU18/02019 funded by MCIN/AEI/10.13039/501100011033 and by "ESF Investing in your future." SG holds a Juan de la Cierva Incorporacion grant (IJC 2020-042749-I) funded by MCIN/AEI/10.13039/501100011033 and the European Union NextGenerationEU/PRTR.Moreno-Giménez, E.; Gandía, M.; Sáez, Z.; Manzanares, P.; Yenush, L.; Orzáez Calatayud, DV.; Marcos, JF.... (2023). FungalBraid 2.0: expanding the synthetic biology toolbox for the biotechnological exploitation of filamentous fungi. Frontiers in Bioengineering and Biotechnology. 11:1-17. https://doi.org/10.3389/fbioe.2023.12228121171

FungalBraid: A GoldenBraid-based modular cloning platform for the assembly and exchange of DNA elements tailored to fungal synthetic biology

[EN] Current challenges in the study and biotechnological exploitation of filamentous fungi are the optimization of DNA cloning and fungal genetic transformation beyond model fungi, the open exchange of ready-to-use and standardized genetic elements among the research community, and the availability of universal synthetic biology tools and rules. The GoldenBraid (GB) cloning framework is a Golden Gate-based DNA cloning system developed for plant synthetic biology through Agrobacterium tumefaciens-mediated genetic transformation (ATMT). In this study, we develop reagents for the adaptation of GB version 3.0 from plants to filamentous fungi through: (i) the expansion of the GB toolbox with the domestication of fungal-specific genetic elements; (ii) the design of fungal-specific GB structures; and (iii) the ATMT and gene disruption of the plant pathogen Penicillium digitatum as a proof of concept. Genetic elements domesticated into the GB entry vector pUPD2 include promoters, positive and negative selection markers and terminators. Interestingly, some GB elements can be directly exchanged between plants and fungi, as demonstrated with the marker hph for Hyg(R) or the fluorescent protein reporter YFP. The iterative modular assembly of elements generates an endless number of diverse transcriptional units and other higher order combinations in the pDGB3 alpha/pDGB3 Omega destination vectors. Furthermore, the original plant GB syntax was adapted here to incorporate specific GB structures for gene disruption through homologous recombination and dual selection. We therefore have successfully adapted the GB technology for the ATMT of fungi. We propose the name of FungalBraid (FB) for this new branch of the GB technology that provides open, exchangeable and collaborative resources to the fungal research community.This work was funded by grants BIO2015-68790-C2-1-R and BIO2016-78601-R from the "Ministerio de Economia y Competitividad" (MINECO, Spain). SG was recipient of a predoctoral scholarship (FPU13/04584) within the FPU program from "Ministerio de Educacion, Cultura y Deporte" (MECD, Spain). We acknowledge the excellent technical assistance of Tania Campos and the help in the microscopy experiments of Jose M. Coll-Marques (IATA, Valencia, Spain). We also thank Dr. Pilar Moya (Universitat Politecnica de Valencia, Spain) for helpful discussions during the initial stages of this project.Hernanz-Koers, M.; Gandía-Gómez, M.; Garrigues-Cubells, SM.; Manzanares-Mir, PM.; Yenush, L.; Orzáez Calatayud, DV.; Marcos -Lopez, JF. (2018). FungalBraid: A GoldenBraid-based modular cloning platform for the assembly and exchange of DNA elements tailored to fungal synthetic biology. Fungal Genetics and Biology. 116:51-61. https://doi.org/10.1016/j.fgb.2018.04.010S516111

El derecho a la protección social de las víctimas de la violencia de género. Estudio sistemático del título II de la L. O. 1/2004, de 28 de diciembre, de protección integral contra la violencia de género de acuerdo con la distribución territorial del Estado

La violencia de género, cuyas víctimas son mujeres en el 90% de los casos, constituye un problema acuciante que afecta a todos los países. Si nos fijamos en la sociedad española, de acuerdo con los datos obtenidos por el barómetro del Centro de Investigaciones Sociológicas de marzo de 2004 (estudio n.º 2.558 ), el 72% de las personas entrevistadas consideraban «muy frecuentes» o «bastan te frecuentes» las agresiones y los maltratos físicos en la pare ja. De hecho, según datos de un informe del Observatorio contra la violencia doméstica y de género del Consejo General del Poder Judicial (2006), hasta junio del año pasa do se celebraron 40.792 juicios por delitos y faltas relacionados con la viol encia contra las mujeres.Investigación financiada mediante subvención recibida de acuerdo con lo previsto en la Orden TAS/1051/2005, de 12 de abril (subvenciones para el Fomento de la Investigación de la Protección Social –FIPROS-

Prevalence of severe esophagitis in Spain. Results of the PRESS study (Prevalence and Risk factors for Esophagitis in Spain: A cross-sectional study)

Background The current prevalence of esophagitis in southern Europe is unknown. In addition, the risk factors for reflux esophagitis are not fully understood. Objective The objective of this article is to assess the prevalence and risk factors for esophagitis in Spain. Methods A prospective, observational, cross-sectional, multicenter study (PRESS study) was conducted among 31 gastrointestinal endoscopy units throughout Spain. A total of 1361 patients undergoing upper gastrointestinal endoscopy were enrolled. Sociodemographic, clinical and treatment data were recorded. Results A total of 95% of patients were Caucasian and 52% were male (mean age: 5317 years). The most frequent symptoms prompting endoscopy were heartburn (40%), regurgitation (26%) and dysphagia (15%). Fifty-four percent of patients undergoing endoscopy were receiving proton pump inhibitor (PPI) treatment. Esophagitis (mainly mild-moderate) was present in 154 (12.4%) patients. The severe form was recorded in only 11 (0.8%) patients. Multivariate analysis results indicated that the likelihood of esophagitis was higher in men (OR=1.91, 95% CI=1.31-2.78), in patients with high GERD-Q scores (OR=1.256, 95% CI=1.176-1.343), weight increase (OR=1.014, 95% CI=1.003-1.025) and high alcohol consumption (OR=2.49, 95% CI=1.16-5.36). Conclusion Severe esophagitis is a rare finding in the Spanish population. Male gender, high GERD-Q score, weight increase and high alcohol consumption are main risk factors for its appearance

The effects of thiopurine therapy on health-related quality of life in Inflammatory Bowel Disease patients

<p>Abstract</p> <p>Background</p> <p>The effect of thiopurine immunomodulators on health-related quality of life (HRQoL) in patients with inflammatory bowel disease (IBD) has been controversial. The aims were to evaluate the HRQoL in patients with IBD treated with thiopurines and assess the short- and long-term impacts of the treatment on HRQoL.</p> <p>Methods</p> <p>Ninety-two consecutive patients who started treatment with thiopurines were prospectively included. Evaluation of HRQoL was performed at months 0, 6, and 12 using two questionnaires, the Short-Form Health Survey (SF-36) and the Inflammatory Bowel Disease Questionnaire (IBDQ).</p> <p>Results</p> <p>Baseline score of IBDQ was 4,6, range (2,31-6,84), with an impairment of the five dimensions of HRQoL compared with inactive patients. Results obtained in 8 dimensions of SF-36 showed worse HRQoL than Spanish general population. At 6 months patients had a significant improvement in overall IBDQ score -5,8 (1,58 -6,97)- and also in all IBDQ dimensions. All the 8 dimensions of SF-36 obtained a significant improvement. At twelve months score of IBDQ was 6,1, range (2,7-6,98), with improvement in all dimensions compared with baseline and 6 months. SF-36 showed a similar significant improvement in all subscales.</p> <p>Conclusions</p> <p>Thiopurine immunomodulators alone or with other treatments have a positive and long lasting impact on HRQoL of IBD patients.</p

Dinámica biliar en periodo interdigestivo y tras la administración de colecistoquinina. Estudio de su fisiología y de su control muscarínico

El aparato digestivo, merced a sus funciones básicas -motora, secretora y absortiva-, es el lugar donde se absorben los nutrientes de la dieta y a través del que se eliminan los productos no asimilables aportados por la ingesta. Asimismo, juega un papel importante más allá de sus límites anatómicos, gracias a su función endocrina. Los productos de la secreción exocrina del páncreas y del hígado, jugo pancreático y bilis respectivamente, son imprescindibles para una adecuada función absortiva. El tracto biliopancreático es un sistema tubular, cuya única función vital es la de servir de elemento de drenaje al intestino, de las secreciones hepática y pancreática. Su actividad motora, como se verá, reside principalmente en la vesícula y en el esfínter de Oddi. Esta actividad, aunque es primordial para conseguir una correcta regulación del vaciamiento, no es imprescindible para la vida, como puede comprobarse en los pacientes a quienes se ha realizado una colecistectomía y/o una esfinterotomía. Sin embargo, un funcionamiento anormal de estas dos estructuras es capaz de generar situaciones patológicas, en ocasiones graves. La prevalencia de la patología biliar benigna es muy alta en todos los países del mundo occidental, y el nuestro no escapa a esta situación general. De esta alta prevalencia se deriva un coste social y un sufrimiento individual considerables, así como unas tasas de mortalidad en absoluto despreciables

Caracterización y modelado de fenómenos de transferencia de calor en un turbocompresor

Consulta en la Biblioteca ETSI Industriales (Riunet)[ES] El mundo del autom ovil est a en continuo desarrollo y evoluci on, es un ente en permanente crecimiento. Nuevas ideas y mejoras surgen de la extensa investigaci on en este campo. En estos tiempos, los factores medio ambientales y econ omicos juegan un papel fundamental en todos los campos de la ciencia, entre los que se incluye de manera notable la automoci on. Para la disminuci on de los impactos de los factores anteriores es clave la disminuci on del consumo de combustible. La sobrealimentaci on es una tecnolog a esencial para el downsizing y el downspeeding del motor de combusti on interna, lo cu al lleva a un disminuci on del consumo. Por ello el inter es en una predicci on m as exacta del comportamiento del turbocompresor ha aumentado considerablemente y en particular la transmisi on de calor. Al tener en cuenta la transmisi on de calor en los modelos de simulaci on se permite separar los efectos aerodin amicos de los efectos de transferencia de calor, ermitiendo el estudio del comportamiento de compresor y turbina de manera separada aumentando as la precisi on de la modelizaci on. Con el modelo de transmisi on de calor a desarrollar en el presente proyecto se pretende incrementar la precisi on en la predicci on de las temperaturas de salida de turbina y compresor, las cu ales son claves en el dise~no del intercooler y los elementos de post-tratamiento. Para todo ello se realizan una serie de ensayos en distintos bancos de ensayos caracterizando t ermicamente distintos turbocompresores.Garzón Garrigues, JV. (2013). Caracterización y modelado de fenómenos de transferencia de calor en un turbocompresor. http://hdl.handle.net/10251/30453.Archivo delegad

Incorporación de la tecnología de impresión 3D al IES Carles Salvador

La tecnología de impresión 3D ofrece la posibilidad de aplicar metodologías activas en los centros. Aporta gran cantidad de beneficios para toda la comunidad educativa (alumnado, familias y profesorado). Esta comunicación pretende explicar el itinerario seguido en el IES Carles Salvador de Aldaia para llevar a cabo dicha incorporación. La formación y acompañamiento al profesorado han sido elementos fundamentales para su éxito. Así como la realización de actividades de formación y motivación entre el alumnado del centro. En este caso, se ha optado por la creación de grupos de expertos entre el alumnado y el profesorado. Estos equipos se encargarán de transmitir y difundir sus experiencias al resto de miembros de la comunidad educativa. Artículo presentado en el 61 Encuentro de Centros Innovadores (Alicante, 22-3-22