Nucleic acids enrichment of fungal pathogens to study host-pathogen interactions

Fungal infections, ranging from superficial to life-threatening infections, represent a major public health problem that affects 25% of the worldwide population. In this context, the study of host-pathogen interactions within the host is crucial to advance antifungal therapy. However, since fungal cells are usually outnumbered by host cells, the fungal transcriptome frequently remains uncovered. We compared three different methods to selectively lyse human cells from in vitro mixes, composed of Candida cells and peripheral blood mononuclear cells. In order to prevent transcriptional modification, the mixes were stored in RNAlater. We evaluated the enrichment of fungal cells through cell counting using microscopy and aimed to further enrich fungal nucleic acids by centrifugation and by reducing contaminant nucleic acids from the host. We verified the enrichment of fungal DNA and RNA through qPCR and RT-qPCR respectively and confirmed that the resulting RNA has high integrity scores, suitable for downstream applications. The enrichment method provided here, i.e., lysis with Buffer RLT followed by centrifugation, may contribute to increase the proportion of nucleic acids from fungi in clinical samples, thus promoting more comprehensive analysis of fungal transcriptional profiles. Although we focused on C. albicans, the enrichment may be applicable to other fungal pathogens

Comparison of the efficiency of different cell lysis methods and different commercial methods for RNA extraction from Candida albicans stored in RNAlater

Background: Obtaining sufficient RNA yield and quality for comprehensive transcriptomic studies is cumbersome for clinical samples in which RNA from the pathogen is present in low numbers relative to the nucleic acids from the host, especially for pathogens, such as yeasts, with a solid cell wall. Therefore, yeast cell lysis including cell wall disruption constitutes an essential first step to maximize RNA yield. Moreover, during the last years, different methods for RNA extraction from yeasts have been developed, ranging from classic hot phenol methods to commercially available specific kits. They offer different RNA yield and quality, also depending on the original storage medium, such as RNAlater. Results: We observed that, for C. albicans cells stored in Tryptic Soy Broth with 15% glycerol, 10min of bead beating in a horizontal position in RiboPure Lysis Buffer provided complete cell lysis. Cell lysis efficiency was decreased to 73.5% when cells were stored in RNAlater. In addition, the RiboPure Yeast Kit (Ambion) offered the highest RNA yield in comparison with the automated platform NucliSENS easyMAG total nucleic extraction (bioMerieux) and the RNeasy Mini Kit (Qiagen) according to NanoDrop and Fragment Analyzer. Moreover, we showed that, in spite of the decrease of cell lysis efficiency after RNAlater storage, as compared to storage in TSB+15% glycerol, RNAlater increased RNA yield during RNA extraction with both RiboPure Yeast Kit and easyMAG, as confirmed by Fragment Analyzer analysis and by RT-qPCR of the RNA from the Internal Transcribed Spacer 2. Conclusions: In our hands, the most efficient cell lysis and highest RNA yield from C. albicans cells stored in RNAlater was obtained by horizontal bead beating in RiboPure Lysis Buffer followed by RNA extraction with the RiboPure Yeast Kit

Propuesta del diseño de un manual de control interno contable del área de Inventario de Suministros en el Hotel Granada Inn, ubicado en el municipio de Granada, Departamento de Granada, durante el II Semestre del año 2016

El presente informe se enfoca en desarrollar una propuesta del diseño de un manual de control interno contable del área de inventario de suministros en el hotel Granada Inn, ubicado en el municipio de Granada, departamento de Granada. Para ello se aplicaron dos entrevistas a los propietarios con el fin de conocer las generalidades y la situación en la que se encuentra el negocio. A través de los instrumentos utilizados, se logró constatar que el hotel Granada Inn se encuentra en la finalización de la primera etapa de construcción por tanto los propietarios carecen de un manual de control interno contable donde se establezcan objetivos, políticas, mecanismos y lineamientos para la operación del área de inventario de suministros. Por tanto elaboramos una propuesta del diseño de un manual de control interno contable al que se sujetará el personal responsable de las operaciones de inventario en el hotel Granada Inn, planteando objetivos general y específicos que servirán de base para el buen funcionamiento del negocio, se establecieron políticas y normas según el modelo Coso I con los cinco componentes característicos de este modelo para controlar las actividades de las empresas, así como mecanismos y lineamientos para la operación de inventario

Exploring the ring-closing metathesis for the construction of the solomonamide macrocyclic core: identification of bioactive precursors

New synthetic strategies directed toward the novel cyclopeptides solomonamides have been explored utilizing an olefin metathesis as the key reaction. In the various strategies investigated, we worked on minimally oxidized systems, and the olefin metathesis reaction demonstrated efficiency and validity for the construction of the macrocyclic core. The described synthetic strategies toward the solomonamides are well suited for the subsequent access to the natural products and represent flexible and diversityoriented routes that allow for the generation of a variety of analogues via oxidative transformations. In addition, preliminary biological evaluations of the generated solomonamide precursors revealed antitumor activity against various tumor cell lines.This work was financially supported by the Ministerio de Economía y Competitividad (MINECO) (ref BIO2014-56092-R, CTQ2014-60223-R and CTQ2016-76311-R) and Junta de Andalucía and “Fondo Europeo de Desarrollo Regional-FEDER” (P12 CTS-1507). I.C.-S. thanks Ministerio de Educación, Cultura y Deporte for a predoctoral fellowship (FPU programme)

Factores asociados a la actividad física y el sedentarismo en los estilos de vida de la juventud española

La comunicación presenta los resultados de un estudio realizado por la Universidad Pablo de Olavide, la Universidad Politécnica de Madrid y la Universidad Europea de Madrid, sobre las tendencias culturales en el abandono de la actividad física y el deporte entre la juventud española. El estudio, que está realizándose actualmente gracias a una ayuda concedida por el Consejo Superior de Deportes (CSD) (Ref. 007/UPB10/12), analiza cómo la adopción de ese nuevo estilo de vida (sedentario) que está progresivamente afianzándose entre la juventud española puede influir de una manera u otra en la percepción subjetiva de la salud y el bienestar entre este grupo de población

Real-time embedded eye detection system

The detection of a person’s eyes is a basic task in applications as important as iris recognition in biometric identification or fatigue detection in driving assistance systems. Current commercial and research systems use software frameworks that require a dedicated computer, whose power consumption, size, and price are significantly large. This paper presents a hardware-based embedded solution for eye detection in real-time. From an algorithmic point-of-view, the popular Viola-Jones approach has been redesigned to enable highly parallel, single-pass image-processing implementation. Synthesized and implemented in an All-Programmable System-on-Chip (AP SoC), this proposal allows us to process more than 88 frames per second (fps), taking the classifier less than 2 ms per image. Experimental validation has been successfully addressed in an iris recognition system that works with walking subjects. In this case, the prototype module includes a CMOS digital imaging sensor providing 16 Mpixels images, and it outputs a stream of detected eyes as 640 × 480 images. Experiments for determining the accuracy of the proposed system in terms of eye detection are performed in the CASIA-Iris-distance V4 database. Significantly, they show that the accuracy in terms of eye detection is 100%.This work has been partially developed within the project RTI2018-099522-B-C4X, funded by the Gobierno de España and FEDER funds, and the ARMORI project (CEIATECH-10) funded by the University of Málaga. Portions of the research in this paper use the CASIA-Iris V4 collected by the Chinese Academy of Sciences - Institute of Automation (CASIA)

Analytical Characterization and Sensory Analysis of Distillates of Di erent Varieties of Grapes Aged by an Accelerated Method

The wine spirits used for the elaboration of Brandy de Jerez are mainly obtained from wines produced from the Airen type of grape, which comes from the vineyards located in the region of La Mancha (Central Spain). This entails a limitation when achieving a product classified as "protected geographic designation". For that purpose, it is necessary that the grape used for the wine spirit comes from the area and not from Castile la Mancha, as has happened until now. Due to this fact, it is necessary to search for a possible alternative grape variety which allows the produced brandy to be eligible for a "protected geographic designation". For that purpose, an accelerated ageing process has been implemented with a method previously optimized to distillates obtained from wines from different varieties of grapes (Airen, Colombard, Corredera, Doradilla, Garrido Fino, Jaen blanco, Moscatel de Alejandria, Palomino Fino, Ugni Blanc, and Zalema) grown in the Jerez Area. They were evaluated, both from the analytical and sensory points of view. The distillates made from Jaen Blanco and Zalema have properties that make them interesting for future development and incorporation into oenological practice

Web based interactive educational software introducing semiconductor laser dynamics: Sound Of Lasers (SOL)

In this work, educational software for intuitive understanding of the basic dynamic processes of semiconductor lasers is presented. The proposed tool is addressed to the students of optical communication courses, encouraging self consolidation of the subjects learned in lectures. The semiconductor laser model is based on the well known rate equations for the carrier density, photon density and optical phase. The direct modulation of the laser is considered with input parameters which can be selected by the user. Different options for the waveform, amplitude and frequency of thpoint. Simulation results are plotted for carrier density and output power versus time. Instantaneous frequency variations of the laser output are numerically shifted to the audible frequency range and sent to the computer loudspeakers. This results in an intuitive description of the “chirp” phenomenon due to amplitude-phase coupling, typical of directly modulated semiconductor lasers. In this way, the student can actually listen to the time resolved spectral content of the laser output. By changing the laser parameters and/or the modulation parameters,consequent variation of the laser output can be appreciated in intuitive manner. The proposed educational tool has been previously implemented by the same authors with locally executable software. In the present manuscript, we extend our previous work to a web based platform, offering improved distribution and allowing its use to the wide audience of the web

Predictive model for polishing times in mould finishing

El pulido, proceso final utilizado en la producción de piezas, es a menudo lento y se aplica manualmente. Normalmente define la rugosidad superficial y la precisión de la pieza. Se lleva a cabo utilizando partículas duras para producir la abrasión de la superficie de la pieza a pulir. En este documento se propone un modelo para predecir el progreso del frente de pulido. El modelo tiene en cuenta la topografía resultante antes de la mecanización a través de su curva de Abbott - Firestone, la velocidad relativa del material abrasivo, la fuerza aplicada, el material de la pieza y el tamaño de los granos abrasivos. Separa las aportaciones realizadas por el material de la pieza y el tipo abrasivo de la topografía previa. También muestra los valores recomendados para los diferentes parámetros de proceso a ser utilizados en la predicción de la evolución del frente de pulido en algunos materiales. Además se estima el tiempo de pulido necesario para eliminar completamente la topografía resultante del fresado utilizando una estrategia de perforación transversal. Por último, se muestra la predicción del modelo del frente de pulido, lo que confirma que el mecanismo de abrasión caracteriza dicho proceso.Polishing, the final process used in the production of parts, is often slow and often applied manually. It normally defines the surface roughness and part accuracy. It is performed using hard particles to produce abrasion of the workpiece surface to be polished. In this paper a model for predicting the progress of the polishing face is proposed. The model considers the topography resulting from prior mechanizing through its Abbott-Firestone curve, the relative speed of the abrasive material, the applied force, the workpiece material and the size of the abrasive grains. It separates the contributions made by the workpiece material and the abrasive type from the prior topography. It also shows recommended values for different process parameters to be used in predicting the evolution of the polishing front in some materials. Additionally, the polishing time required to completely remove the topography that results from ball milling using a cross drilling strategy is estimated. Finally, the model's correct prediction of the polished front is shown, confirming that the abrasion mechanism characterizes the sandpaper polishing process.Peer Reviewe