Person: Prieto Prieto, Antonio Daniel
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First Name
Antonio Daniel
Last Name
Prieto Prieto
Affiliation
Universidad Complutense de Madrid
Faculty / Institute
Farmacia
Department
Microbiología y Parasitología
Area
Microbiología
Identifiers
13 results
Search Results
Now showing 1 - 10 of 13
- Project number: 369
Item Dinamizando el aula: implementación de Wooclap en la docencia de Microbiología e Inmunología, y su utilidad en aprendizaje significativo(2024) Román González, Elvira; Pla Alonso, Jesús; Navarro García, Federico; Prieto Prieto, Antonio Daniel; García Pastor, Lucía; Blesa Esteban, Alba Mercedes Item Cooperative Role of MAPK Pathways in the Interaction of Candida albicans with the Host Epithelium(Microorganisms, 2019) Correia, C. Inês Ribeiro; Prieto Prieto, Antonio Daniel; Román García, Elvira; Wilson, Duncan; Hube, Bernhard; Alonso Monge, Rebeca María Del Mar; Pla Alonso, JesúsCandida albicans is an important human fungal pathogen responsible for tens of millions of infections as well as hundreds of thousands of severe life-threatening infections each year. MAP kinase (MAPK) signal transduction pathways facilitate the sensing and adaptation to external stimuli and control the expression of key virulence factors such as the yeast-to-hypha transition, the biogenesis of the cell wall, and the interaction with the host. In the present study, we have combined molecular approaches and infection biology to analyse the role of C. albicans MAPK pathways during an epithelial invasion. Hog1 was found to be important for adhesion to abiotic surfaces but was dispensable for damage to epithelial cells. The Mkc1 cell wall integrity (CWI) and Cek1 pathways, on the other hand, were both required for oral epithelial damage. Analysis of the ability to penetrate nutrient-rich semi-solid media revealed a cooperative role for Cek1 and Mkc1 in this process. Finally, cek2∆ (as well as cek1∆) but not mkc1∆ or hog1∆ mutants, exhibited elevated β-glucan unmasking as revealed by immunofluorescence studies. Therefore, the four MAPK pathways play distinct roles in adhesion, epithelial damage, invasion and cell wall remodelling that may contribute to the pathogenicity of C. albicans.Item The HOG MAPK pathway in Candida albicans: more than an osmosensing pathway.(International microbiology: the official journal of the Spanish Society for Microbiology, 2019) Román, Elvira; Correia, C. Inês Ribeiro; Alonso Monge, Rebeca María Del Mar; Prieto Prieto, Antonio Daniel; Pla Alonso, JesúsIn 1993, Brewster and Gustin described the existence of a kinase whose activity was essential for Saccharomyces cerevisiae to grow in environments with high osmolarity. This led to the discovery of the HOG pathway, a MAP kinase (MAPK) pathway that has been revealed to be crucial to respond to a wide range of stress conditions frequently encountered by fungi in their common habitats. MAPK signaling is initiated at the plasma membrane, where triggering stimuli lead to a phosphorylation cascade that ultimately activates transcription factors to ensure an appropriate adaptive response. In pathogenic fungi, the HOG pathway gains special significance as it is involved in traits related to pathogenicity; these include biofilm formation, adhesion to surfaces, and morphogenetic and epigenetic transitions. It also plays a role in controlling both the pathogen and the commensal state program. Understanding the signals leading to its activation, the elements of the pathways and the targets of the pathway are therefore of primary importance in the design of novel antifungals.- Project number: 199
Item Propuesta de intervención educativa centrada en el alumno mediante el modelo de clase invertida en la docencia práctica de la asignatura Microbiología Clínica(2021) Román González, Elvira; Pla Alonso, Jesús; Jiménez Cid, Víctor; Fernández-Acero Bascones, Teresa; Prieto Prieto, Antonio Daniel; Rodríguez Escudero, María Isabel; Hidalgo Vico, Susana; Garcia Lavilla, Beatriz; González Rubio, Gema; Pavón Verges, Mónica; Valentí Sanguino, Marta; Román González, Marcos; Sánchez Angulo, ManuelHemos implementado la metodología de clase invertida como modelo de aprendizaje centrado en el alumno en la docencia práctica de la asignatura Microbiología Clínica con el objetivo dinamizar y optimizar el tiempo en el aula. Este tipo de docencia ha favorecido la participación activa del estudiante, su aprendizaje y fomentado un mayor pensamiento crítico que permite la toma de decisiones durante la consecución de las prácticas. Item Deletion of the SKO1 Gene in a hog1 Mutant Reverts Virulence in Candida albicans(Journal of Fungi, 2019) Urrialde, Verónica; Prieto Prieto, Antonio Daniel; Hidalgo Vico, Susana; Román González, Elvira; Pla Alonso, Jesús; Alonso Monge, Rebeca María Del MarCandida albicans displays the ability to adapt to a wide variety of environmental conditions, triggering signaling pathways and transcriptional regulation. Sko1 is a transcription factor that was previously involved in early hypoxic response, cell wall remodeling, and stress response. In the present work, the role of sko1 mutant in in vivo and ex vivo studies was explored. The sko1 mutant behaved as its parental wild type strain regarding the ability to colonize murine intestinal tract, ex vivo adhesion to murine gut epithelium, or systemic virulence. These observations suggest that Sko1 is expendable during commensalism or pathogenesis. Nevertheless, the study of the hog1 sko1 double mutant showed unexpected phenotypes. Previous researches reported that the deletion of the HOG1 gene led to avirulent C. albicans mutant cell, which was, therefore, unable to establish as a commensal in a gastrointestinal murine model. Here, we show that the deletion of sko1 in a hog1 background reverted the virulence of the hog1 mutant in a systemic infection model in Galleria mellonella larvae and slightly improved the ability to colonize the murine gut in a commensalism animal model compared to the hog1 mutant. These results indicate that Sko1 acts as a repressor of virulence related genes, concluding that Sko1 plays a relevant role during commensalism and systemic infection.- Project number: 265
Item SWI@UCM 2.0: Consolidación de Small World Initiative: descubrimiento y uso racional de antibióticos mediante aprendizaje-servicio en la Comunidad de Madrid(2018) Jiménez Cid, Víctor; Rodríguez Escudero, María Isabel; Díez Orejas, Rosalía María; Molina Martín, María; Rodríguez Fernández, Carmina; Navarro García, Federico; Arroyo Nombela, Francisco Javier; Román González, Elvira; Martín Brieva, Humberto; Fernández-Acero Bascones, Teresa; Sanz Santamaría, Ana Belén; Díaz Del Toro, Silvia; Calvo De Pablo, Pilar; Patiño Álvarez, Aurora Belén; González Zorn, Bruno; Suárez Rodríguez, Mónica; Goyache Goñi, Joaquín; Escudero García-Calderón, José Antonio; Prieto Prieto, Antonio Daniel; Ugarte Ruiz, María; Gil Serna, JessicaDurante el curso anterior un equipo de la UCM en el marco de un proyecto previo INNOVA-Docencia UCM instauró en España de manera pionera la iniciativa de aprendizaje activo Small World Initiative, de origen norteamericano. Los objetivos de esta iniciativa son (1) Crear cultura científica y acercar la investigación biomédica a niveles educativos en los que los estudiantes tienen aún capacidad de decisión sobre su futura orientación formativa con el fin de fomentar la vocación en I+D; y (2) Promover la concienciación social sobre el uso racional de los antibióticos y la amenaza de la resistencia bacteriana a estos fármacos. En el entorno español se propuso implementar esta estrategia mediante Aprendizaje-Servicio. En esta segunda edición (SWI@UCM 2.0) se ha trabajado en la consolidación, expansión y mejora del proyecto, con énfasis en la integración de los diversos niveles educativos que integran el proyecto (universitario y preuniversitario) y la divulgación científica. - Project number: 40
Item SWI@UCM: Implantación en España de la Small World Initiative (descubrimiento de antibióticos por “crowdsourcing”) mediante una estrategia de aprendizaje-servicio(2017) Jiménez Cid, Víctor; González Zorn, Bruno; Valderrama Conde, María José; Calvo De Pablo, Pilar; Navarro García, Federico; Molina Martín, María; Pla Alonso, Jesús; Fernández-Acero Bascones, Teresa; Díez Orejas, Rosalía María; Rodríguez Fernández, Carmina; Román González, Elvira; Martín Brieva, Humberto; Sanz Santamaría, Ana Belén; Prieto Prieto, Antonio Daniel; Arregui García-Roves, Lucía; Vázquez Estévez, María Covadonga Inmaculada; Patiño Álvarez, Aurora Belén; Silóniz Jiménez, María Isabel De; Gil Serna, Jessica; Suárez Rodríguez, Mónica; Juan Ferré, Lucía De; Thomas López, Daniel; Escudero García-Calderón, José AntonioSmall World Initiative (SWI) es un proyecto internacional cuyos objetivos son: (1) acercar la cultura científica y la investigación biomédica a niveles educativos en los que los estudiantes tienen aún capacidad de decisión sobre su futura orientación formativa con el fin de fomentar la vocación en I+D; y (2) divulgar una de las líneas prioritarias marcadas por la Organización Mundial de la Salud (OMS), que es la concienciación social sobre el uso racional de los antibióticos y la amenaza de la resistencia bacteriana. Para lograr estos objetivos SWI desarrolla un proyecto real de investigación dirigido al descubrimiento de nuevos antibióticos mediante una estrategia de “crowdsourcing”, de modo que la comunidad participa de forma activa en las tareas de investigación. En esta experiencia piloto de implantación en España de esta iniciativa, SWI@UCM hemos optado por una estrategia basada en aprendizaje-servicio integrando dos niveles educativos, de modo que los estudiantes universitarios, como parte de su formación, son responsables de la gestión y organización de laboratorios de microbiología en institutos de educación secundaria y bachillerato, dirigiendo a los jóvenes científicos en estudio de la diversidad microbiana en los suelos para el descubrimiento de microorganismos productores de antibióticos potencialmente nuevos. Item Identification of Clinical Isolates of Candida albicans with Increased Fitness in Colonization of the Murine Gut(Journal of Fungi, 2021) Alonso Monge, Rebeca María Del Mar; Prieto Prieto, Antonio Daniel; Coman, Ioana V.; Rochas, Sara; Arana, David M.; Hidalgo Vico, Susana; Pla Alonso, Jesús; Román González, ElviraThe commensal and opportunistic pathogen Candida albicans is an important cause of fungal diseases in humans, with the gastrointestinal tract being an important reservoir for its infections. The study of the mechanisms promoting the C. albicans commensal state has attracted considerable attention over the last few years, and several studies have focused on the identification of the intestinal human mycobiota and the characterization of Candida genes involved in its establishment as a commensal. In this work, we have barcoded 114 clinical C. albicans isolates to identify strains with an enhanced fitness in a murine gastrointestinal commensalism model. The 114 barcoded clinical isolates were pooled in four groups of 28 to 30 strains that were inoculated by gavage in mice previously treated with antibacterial therapy. Eight strains that either exhibited higher colonization load and/or remained in the gut after antibiotic removal were selected. The phenotypic analysis of these strains compared to an RFP-tagged SC5314 wild type strain did not reveal any specific trait associated with its increased colonization; all strains were able to filament and six of the eight strains displayed invasive growth on Spider medium. Analysis of one of these strains, CaORAL3, revealed that although mice required previous bacterial microbiota reduction with antibiotics to be able to be colonized, removal of this procedure could take place the same day (or even before) Candida inoculation. This strain was able to colonize the intestine of mice already colonized with Candida without antibiotic treatment in co-housing experiments. CaORAL3 was also able to be established as a commensal in mice previously colonized by another (CaHG43) or the same (CaORAL3) C. albicans strain. Therefore, we have identified C. albicans isolates that display higher colonization load than the standard strain SC5314 which will surely facilitate the analysis of the factors that regulate fungal colonization.Item Implementation of a CRISPR-Based System for Gene Regulation in Candida albicans.(mSphere, 2019) Román González, Elvira; Coman, Ioana V.; Prieto Prieto, Antonio Daniel; Alonso Monge, Rebeca María Del Mar; Pla Alonso, JesúsClustered regularly interspaced short palindromic repeat (CRISPR) methodology is not only an efficient tool in gene editing but also an attractive platform to facilitate DNA, RNA, and protein interactions. We describe here the implementation of a CRISPR-based system to regulate expression in the clinically important yeast By fusing an allele of Cas9 devoid of nuclease activity to a transcriptional repressor (Nrg1) or activator (Gal4), we were able to show specific repression or activation of the tester gene , encoding the cytosolic catalase. We generated strains where a 1.6-kbp upstream regulatory region of controls the expression of the green fluorescent protein (GFP) and demonstrated the functionality of the constructs by quantitative PCR (qPCR), flow cytometry, and analysis of sensitivity/resistance to hydrogen peroxide. Activation and repression were strongly dependent on the position of the complex in this regulatory region. We also improved transcriptional activation using an RNA scaffolding strategy to allow interaction of inactive variants of Cas9 (dCas9) with the RNA binding protein MCP (monocyte chemoattractant protein) fused to the VP64 activator. The strategy shown here may facilitate the analysis of complex regulatory traits in this fungal pathogen. CRISPR technology is a new and efficient way to edit genomes, but it is also an appealing way to regulate gene expression. We have implemented CRISPR as a gene expression platform in using fusions between a Cas9 inactive enzyme and specific repressors or activators and demonstrated its functionality. This will allow future manipulation of complex virulence pathways in this important fungal pathogen.Item The Pho4 transcription factor mediates the response to arsenate and arsenite in Candida albicans.(Frontiers in microbiology, 2015) Urrialde, Verónica; Prieto Prieto, Antonio Daniel; Pla Alonso, Jesús; Alonso Monge, Rebeca María Del MarArsenate (As (V)) is the dominant form of the toxic metalloid arsenic (As). Microorganisms have consequently developed mechanisms to detoxify and tolerate this kind of compounds. In the present work, we have explored the arsenate sensing and signaling mechanisms in the pathogenic fungus Candida albicans. Although mutants impaired in the Hog1 or Mkc1-mediated pathways did not show significant sensitivity to this compound, both Hog1 and Mkc1 became phosphorylated upon addition of sodium arsenate to growing cells. Hog1 phosphorylation upon arsenate challenge was shown to be Ssk1-dependent. A screening designed for the identification of transcription factors involved in the arsenate response identified Pho4, a transcription factor of the myc-family, as pho4 mutants were susceptible to As (V). The expression of PHO4 was shortly induced in the presence of sodium arsenate in a Hog1-independent manner. Pho4 level affects Hog1 phosphorylation upon As (V) challenge, suggesting an indirect relationship between Pho4 activity and signaling in C. albicans. Pho4 also mediates the response to arsenite as revealed by the fact that pho4 defective mutants are sensitive to arsenite and Pho4 becomes phosphorylated upon sodium arsenite addition. Arsenite also triggers Hog1 phosphorylation by a process that is, in this case, independent of the Ssk1 kinase. These results indicate that the HOG pathway mediates the response to arsenate and arsenite in C. albicans and that the Pho4 transcription factor can differentiate among As (III), As (V) and Pi, triggering presumably specific responses.