TUP1-mediated filamentation in Candida albicans leads to inability to colonize the mouse gut.

AIM To investigate the role of Candida albicans TUP1-mediated filamentation in the colonization of the mice gut. MATERIALS & METHODS We used molecular genetics to generate a strain where filamentation is regulated by altering the expression of the TUP1 gene with tetracyclines. RESULTS The colonization rates reached with the TUP1-RFP strain were lower compared with wild-type strain and completely absent after induction of filamentation. No differences in the susceptibility to bile salts nor in the adhesion to the mouse intestine epithelium were observed. CONCLUSION Blockage of C. albicans in a filamentous form impedes gut cell colonization in the mouse

Candida albicans: genetics, dimorphism and pathogenicity

Candida albicans is a dimorphic fungus that causes severe opportunistic infections in humans. Recent advances in molecular biology techniques applied to this organism (transformation systems, gene disruption strategies, new reporter systems, regulatable promoters) allow a better knowledge of both the molecular basis of dimorphism and the role of specific genes in Candida morphogenesis. These same molecular approaches together with the development of appropriate experimental animal models to analyze the virulence of particular mutants, may help to understand the molecular basis of Candida virulence

De la “lección magistral” al Aprendizaje Basado en Proyectos (ABP) "personalizado" a las características del alumnado de Ciencias de la Salud

En el presente Proyecto llevamos a cabo la implantación del Aprendizaje Basado en Proyectos (ABP) en el aula universitaria utilizando la asignatura de Microbiología del Grado en Farmacia, en Ciencia y Tecnología de los Alimentos, y en Óptica y Optometría. En cada uno de dichos Grados se comparó esta metodología con la tradicional clase magistral y además se adaptó el ABP en los tres Grados, según la idiosincrasia del alumnado en cada uno de ellos, constituyendo lo que llamamos ABP “personalizado”

Educating in antimicrobial resistance awareness: adaptation of the Small World Initiative program to service-learning.

The Small World Initiative (SWI) and Tiny Earth are a consolidated and successful education programs rooted in the USA that tackle the antibiotic crisis by a crowdsourcing strategy. Based on active learning, it challenges young students to discover novel bioactive-producing microorganisms from environmental soil samples. Besides its pedagogical efficiency to impart microbiology content in academic curricula, SWI promotes vocations in research and development in Experimental Sciences and, at the same time, disseminates the antibiotic awareness guidelines of the World Health Organization. We have adapted the SWI program to the Spanish academic environment by a pioneering hierarchic strategy based on service-learning that involves two education levels (higher education and high school) with different degrees of responsibility. Throughout the academic year, 23 SWI teams, each consisting of 3-7 undergraduate students led by one faculty member, coordinated off-campus programs in 22 local high schools, involving 597 high school students as researchers. Post-survey-based evaluation of the program reveals a satisfactory achievement of goals: acquiring scientific abilities and general or personal competences by university students, as well as promoting academic decisions to inspire vocations for science- and technology-oriented degrees in younger students, and successfully communicating scientific culture in antimicrobial resistance to a young stratum of society

Differential effects of graphene oxide nanosheets on Candida albicans phagocytosis by murine peritoneal macrophages

Macrophages, as effector cells involved in the innate and adaptive immunity, play a key role in the response to nanomaterials as graphene oxide (GO) and in their cellular uptake. The interactions at the interface of GO nanosheets, macrophages and microbial pathogens need to be assessed to determine the possible impairment of the immune system induced by biomedical treatments with this nanomate-rial. Here, we have evaluated by flow cytometry and confocal microscopy the ability of murine peritoneal macrophages to phagocytose the fungal pathogen Candida albicans, alive or heat-killed, after treatment with poly(ethylene glycol-amine)-derivatized GO nanosheets (PEG-GO). After GO treatment, differences in fungal phagocytosis were observed between macrophages that had taken up GO nanosheets (GO+ pop-ulation) and those that had not (GO population). GO treatment increased the ingested alive yeasts in GO macrophages, whereas phagocytosis diminished in the GO+ population. Ingestion of heat-killed yeasts was slightly higher in both GO and GO+ populations when comparing with control macrophages. For the first time, we show that GO uptake by macrophages modulates its phagocytic capability, affecting differentially the subsequent ingestion of either alive or heat-killed yeasts. Enhanced ingestion of heat-killed yeast by GO-treated macrophages suggests a beneficial role of this nanomaterial for the clearance of dead microorganisms during infection.Ministerio de Economía y Competitividad(España)Fundação para a Ciência e a Tecnologia(Portugal)Depto. de Química en Ciencias FarmacéuticasFac. de FarmaciaTRUEpu

Immunoproteomic analysis of the protective response obtained from vaccination with Candida albicans ecm33 cell wall mutant in mice

Systemic candidiasis remains a major cause of disease and death, particularly among immunocompromised patients. The cell wall of Candida albicans defines the interface between host and pathogen and surface proteins are major elicitors of host immune responses during candidiasis. The C. albicans ecm33 mutant (RML2U) presents an altered cell wall, which entails an increase in the outermost protein layer. Vaccination of BALB/c mice with RML2U mutant protected them from a subsequent lethal infection with virulent strain SC5314 in a systemic candidiasis model. Using immunoproteomics (2-DE followed by Immunoblotting) we detected 29 immunoreactive proteins specifically recognized by antibodies from vaccinated mice sera, six of which are described as immunogenic for the first time (Gnd1p, Cit1p, Rpl10Ep, Yst1p, Cys4p, Efb1p). Furthermore, identification of wild type and mutant cell surface proteome (surfome), confirmed us that the mutant surfome presented a larger number of proteins than the wild type. Interestingly, proteins exclusively identified in the mutant surfome (Met6p, Eft2p, Tkl1p, Rpl10Ep, Atp1p, Atp2p) were also detected as immunogenic, supporting the idea that their surface location enhances their immunoprotective capacity.Comunidad Autónoma de MadridUniversidad Complutense de MadridComisión Interministerial de Ciencia y TecnologíaDepto. de Microbiología y ParasitologíaFac. de FarmaciaTRUEpu

Promising biomedical subcutaneous delivery system in opioid disaccustom process: In vitro/in vivo evaluation of naloxone microparticles on antagonist effect of morphine

The addiction induced by the misuse of opioids, is not only a public health emergency but also a social and economic welfare. The main therapy is based on opioid antagonists. Oral and injectable naltrexone administration is the most widely used, presenting some inconveniences: poor patient adherence to the oral daily dosing schedule, cases of hepatitis and clinically significant liver dysfunction. This study proposes the in vitro e in vivo evaluation of anti-opioid properties of naloxone loaded-poly(lactic-co-glycolic) acid microparticles (NX-MP). In previous studies, NX-MP showed in vitro sustained naloxone release for one week at least. Our results demonstrate the in vitro efficacy of the NX-MP antagonizing for 7 days the morphine effect in SH-SY5Y cells and myenteric plexus-longitudinal muscle preparations isolated from guinea-pig ileum. The in vivo evaluation of the NX-MP was carried out in mice testing the antagonism of the antinociceptive effect of morphine. Results showed that subcutaneous administration of NX-MP blocked the morphine effect. The results of this work suggest that the subcutaneous administration of NX-MP enhances naloxone therapeutic efficacy as non-addictive medication and could be a promising alternative to naltrexone. Furthermore, the dose of NX-MP can be adapted to the patient necessities. It would be an interesting advantage to treat opioid-addiction.This research was partially funded by Universidad Complutense de Madrid Research Group (910939) and by Plan Nacional sobre Drogas, Gobierno de Espan