Identification of yeast genes that confer resistance to chitosan oligosaccharide (COS) using chemogenomics

Background: Chitosan oligosaccharide (COS), a deacetylated derivative of chitin, is an abundant, and renewable natural polymer. COS has higher antimicrobial properties than chitosan and is presumed to act by disrupting/permeabilizing the cell membranes of bacteria, yeast and fungi. COS is relatively non-toxic to mammals. By identifying the molecular and genetic targets of COS, we hope to gain a better understanding of the antifungal mode of action of COS. Results: Three different chemogenomic fitness assays, haploinsufficiency (HIP), homozygous deletion (HOP), and multicopy suppression (MSP) profiling were combined with a transcriptomic analysis to gain insight in to the mode of action and mechanisms of resistance to chitosan oligosaccharides. The fitness assays identified 39 yeast deletion strains sensitive to COS and 21 suppressors of COS sensitivity. The genes identified are involved in processes such as RNA biology (transcription, translation and regulatory mechanisms), membrane functions (e.g. signalling, transport and targeting), membrane structural components, cell division, and proteasome processes. The transcriptomes of control wild type and 5 suppressor strains overexpressing ARL1, BCK2, ERG24, MSG5, or RBA50, were analyzed in the presence and absence of COS. Some of the up-regulated transcripts in the suppressor overexpressing strains exposed to COS included genes involved in transcription, cell cycle, stress response and the Ras signal transduction pathway. Down-regulated transcripts included those encoding protein folding components and respiratory chain proteins. The COS-induced transcriptional response is distinct from previously described environmental stress responses (i.e. thermal, salt, osmotic and oxidative stress) and pre-treatment with these well characterized environmental stressors provided little or any resistance to COS. Conclusions: Overexpression of the ARL1 gene, a member of the Ras superfamily that regulates membrane trafficking, provides protection against COS-induced cell membrane permeability and damage. We found that the ARL1 COS-resistant over-expression strain was as sensitive to Amphotericin B, Fluconazole and Terbinafine as the wild type cells and that when COS and Fluconazole are used in combination they act in a synergistic fashion. The gene targets of COS identified in this study indicate that COS’s mechanism of action is different from other commonly studied fungicides that target membranes, suggesting that COS may be an effective fungicide for drug-resistant fungal pathogens.

Análisis experimental de la pérdida de adherencia hormigón-acero en hormigones sometidos a altas temperaturas

En el conjunto de materiales de construcción habituales en la edificación y las obras de ingeniería, el hormigón destaca entre otras razones por su excelente comportamiento frente a las altas temperaturas y la exposición al fuego. El presente estudio se centra en la adherencia residual entre el hormigón y las barras de acero corrugado soldable tras exponer probetas a altas temperaturas y enfriarlas hasta temperatura ambiente por convección natural. El estudio incluye hormigones de resistencia convencional, hormigones de alta resistencia y hormigones reforzados con fibras de polipropileno y fibras de acero. La adherencia hormigón-acero se ha medido mediante el conocido ensayo de pull-out. La campaña experimental también ha incluido la resistencia a compresión y la resistencia a tracción indirecta. Parte de las probetas se han ensayado a 28 días de edad a temperatura ambiente. A 60 días de edad se han repetido los ensayos a temperatura ambiente y se han realizado esos mismos ensayos en probetas calentadas en un horno industrial hasta tres rangos de temperatura: 450°C, 650°C y 825°C. Previo al proceso de calentamiento han sido sometidas durante 3 horas a un escalón de secado a 120°C. Mediante la metodología propuesta ha sido posible caracterizar la evolución de la pérdida de adherencia residual entre el acero y el hormigón conforme se exponen los especímenes a temperaturas más elevadas. La adición de fibras no tiene una influencia clara en la adherencia a temperatura ambiente. Sin embargo, sí se ha conseguido determinar una mejora sustancial de la adherencia residual en los hormigones, reforzados con fibras de acero sometidos a altas temperaturas.

Características del desarrollo de una mirada profesional en estudiantes para profesor de matemáticas en un contexto b-learning

Esta investigación presenta un estudio cuyo objetivo es identificar aspectos que apoyan el desarrollo de la mirada profesional en estudiantes para profesores de matemáticas en un contexto b-learning. Analizamos las producciones de un grupo de estudiantes para profesores de matemáticas de educación secundaria (documentos escritos y participaciones en un debate on-line) cuando analizaban el razonamiento proporcional de estudiantes de educación secundaria. Los resultados indican que la interacción en el debate en línea permitió a algunos estudiantes para profesor mejorar su capacidad de identificar e interpretar aspectos relevantes del pensamiento matemático de los estudiantes de educación secundaria. Estos resultados indican que el desarrollo de “una mirada profesional” del profesor es un proceso complicado pero que la posibilidad de construir un discurso progresivo en línea es un factor importante para su desarrollo.

Gaia-ESO Survey: Analysis of pre-main sequence stellar spectra

Context. The Gaia-ESO Public Spectroscopic Survey is obtaining high-quality spectroscopy of some 100 000 Milky Way stars using the FLAMES spectrograph at the VLT, down to V = 19 mag, systematically covering all the main components of the Milky Way and providing the first homogeneous overview of the distributions of kinematics and chemical element abundances in the Galaxy. Observations of young open clusters, in particular, are giving new insights into their initial structure, kinematics, and their subsequent evolution. Aims. This paper describes the analysis of UVES and GIRAFFE spectra acquired in the fields of young clusters whose population includes pre-main sequence (PMS) stars. The analysis is applied to all stars in such fields, regardless of any prior information on membership, and provides fundamental stellar atmospheric parameters, elemental abundances, and PMS-specific parameters such as veiling, accretion, and chromospheric activity. Methods. When feasible, different methods were used to derive raw parameters (e.g. line equivalent widths) fundamental atmospheric parameters and derived parameters (e.g. abundances). To derive some of these parameters, we used methods that have been extensively used in the past and new ones developed in the context of the Gaia-ESO survey enterprise. The internal precision of these quantities was estimated by inter-comparing the results obtained by these different methods, while the accuracy was estimated by comparison with independent external data, such as effective temperature and surface gravity derived from angular diameter measurements, on a sample of benchmarks stars. A validation procedure based on these comparisons was applied to discard spurious or doubtful results and produce recommended parameters. Specific strategies were implemented to resolve problems of fast rotation, accretion signatures, chromospheric activity, and veiling. Results. The analysis carried out on spectra acquired in young cluster fields during the first 18 months of observations, up to June 2013, is presented in preparation of the first release of advanced data products. These include targets in the fields of the ρ Oph, Cha I, NGC 2264, γ Vel, and NGC 2547 clusters. Stellar parameters obtained with the higher resolution and larger wavelength coverage from UVES are reproduced with comparable accuracy and precision using the smaller wavelength range and lower resolution of the GIRAFFE setup adopted for young stars, which allows us to provide stellar parameters with confidence for the much larger GIRAFFE sample. Precisions are estimated to be ≈120 K rms in Teff, ≈0.3 dex rms in log g, and ≈0.15 dex rms in [Fe/H] for the UVES and GIRAFFE setups.