Bioethics and health law: the living will. Proposal to create a living will record in Europe

In this piece of work, we present a proposal to create a living will record in Europe that will allow people to have the same rights wherever they are. Therefore, this article will be studied from the bioethics and health law theory

Nitric oxide in fungi: is there NO light at the end of the tunnel?

Nitric oxide (NO) is a remarkable gaseous molecule with multiple and important roles in different organisms, including fungi. However, the study of the biology of NO in fungi has been hindered by the lack of a complete knowledge on the different metabolic routes that allow a proper NO balance, and the regulation of these routes. Fungi have developed NO detoxification mechanisms to combat nitrosative stress, which have been mainly characterized by their connection to pathogenesis or nitrogen metabolism. However, the progress on the studies of NO anabolic routes in fungi has been hampered by efforts to disrupt candidate genes that gave no conclusive data until recently. This review summarizes the different roles of NO in fungal biology and pathogenesis, with an emphasis on the alternatives to explain fungal NO production and the recent findings on the involvement of nitrate reductase in the synthesis of NO and its regulation during fungal development

High-throughput format for the phenotyping of fungi on solid substrates

Filamentous fungi naturally grow on solid surfaces, yet most genetic and biochemical analyses are still performed in liquid cultures. Here, we report a multiplexing platform using high-throughput photometric continuous reading that allows parallel quantification of hyphal growth and reporter gene expression directly on solid medium, thereby mimicking natural environmental conditions. Using this system, we have quantified fungal growth and expression of secondary metabolite GFP-based reporter genes in saprophytic Aspergillus and phytopathogenic Fusarium species in response to different nutrients, stress conditions and epigenetic modifiers. With this method, we provide not only novel insights into the characteristic of fungal growth but also into the metabolic and time-dependent regulation of secondary metabolite gene expression

Variance prediction for population size estimation

Design unbiased estimation of population size by stereological methods is an efficient alternative to automatic computer vision methods, which are generally biased. Moreover, stereological methods offer the possibility of predicting the error variance from a single sample. Here we explore the statistical performance of two alternative variance estimators on a dataset of 26 labelled crowd pictures. The empirical mean square errors of the variance predictors are compared by means of Monte Carlo resampling

Evaluating the pharmacological response in fluorescence microscopy images: The Δm algorithm

Current drug discovery procedures require fast and effective quantification of the pharmacological response evoked in living cells by agonist compounds. In the case of G-protein coupled receptors (GPCRs), the efficacy of a particular drug to initiate the endocytosis process is related to the formation of endocytic vesicles or endosomes and their subsequent internalisation within intracellular compartments that can be observed with high spatial and temporal resolution by fluorescence microscopy techniques. Recently, an algorithm has been proposed to evaluate the pharmacological response by estimating the number of endosomes per cell on time series of images. However, the algorithm was limited by the dependence on some manually set parameters and in some cases the quality of the image does not allow a reliable detection of the endosomes. Here we propose a simple, fast and automated image analysis method?the ?m algorithm- to quantify a pharmacological response with data obtained from fluorescence microscopy experiments. This algorithm does not require individual object detection and computes the relative increment of the third order moment in fluorescence microscopy images after filtering with the Laplacian of Gaussian function. It was tested on simulations demonstrating its ability to discriminate different experimental situations according to the number and the fluorescence signal intensity of the simulated endosomes. Finally and in order to validate this methodology with real data, the algorithm was applied to several time-course experiments based on the endocytosis of the mu opioid receptor (MOP) initiated by different agonist compounds. Each drug displayed a different ?m sigmoid time-response curve and statistically significant differences were observed among drugs in terms of efficacy and kinetic parameters

Estudio morfológico, isotópico y proteómico de la fauna del Pleistoceno y Holoceno de Cova dos Santos (Abadín, Lugo, NO España)

[Abstract] Cova dos Santos is a karstic cavity in Abadín (Lugo), in a hitherto unexplored area that may have been the natural route between the well-known Quaternary faunas of the Cantabrian Mountain Range and those located further south in Galicia, such as in the Serra do Courel. The surface surveys carried out during the topographic layout revealed the presence of deposits of bone remains, usually extremely fragmented, of medium and large vertebrates. Due to the nature of these remains, different molecular techniques (ZooMS, stable isotopes), radiocarbon dating, and morphological and metric analysis were used to characterise the remains present at the site. Combining these methods, it has been possible to identify different taxa such as Ursus speleaeus, Ursus arctos, Panthera pardus, Cervus elaphus, Rhinocerotidae, and to confirm the occupation of this cave since at least 43000 years ago calBP. The presence of domestic species, such as Ovis aries, Equus sp. and Gallus gallus, also shows the use of this cave in more recent times.[Resumen] Cova dos Santos es una cavidad kárstica en Abadín (Lugo), en una zona hasta ahora inexplorada que podría haber sido la ruta natural entre las conocidas faunas cuaternarias de la cordillera Cantábrica y las situadas más al sur de Galicia, como en la Serra do Courel. La prospección de superficie realizada durante los trabajos de trazado topográfico reveló la presencia de depósitos de restos óseos, generalmente muy fragmentados, pertenecientes a vertebrados de mediano y gran tamaño. Debido al carácter de estos restos, se emplearon distintas técnicas moleculares (ZooMS, isótopos estables), dataciones por radiocarbono, y análisis morfológico y métrico para poder caracterizar los restos presentes en el yacimiento. Combinando estos métodos, se han podido identificar distintos taxones como Ursus speleaeus, Ursus arctos, Panthera pardus, Cervus elaphus, Rhinocerotidae, y constatar la ocupación de esta cueva desde hace al menos 43.000 años calBP. La presencia de especies domésticas, como Ovis aries, Equus sp. o Gallus gallus, también pone de manifiesto el uso de esta cavidad en épocas más recientes.This research was supported by financial assistance from the Sociedad Española de Paleontología ID AJISEP-2019-07121 and funds from the Instituto Universitario de Xeoloxía of the UDC. A. García-Vázquez is supported by a postdoctoral ICUB Fellowship for Young Researchers of the University of BucharestSociedad Española de Paleontología; AJISEP-2019-0712

Evidence for an arginine-dependent route for the synthesis of NO in the model filamentous fungus Aspergillus nidulans

Nitric oxide (NO) is a signalling molecule in eukaryotic and prokaryotic organisms. NO levels transiently boost upon induction of conidiation in Aspergillus nidulans. Only one pathway for NO synthesis involving nitrate reductase has been reported in filamentous fungi so far, but this does not satisfy all the NO produced in fungal cells. Here we provide evidence for at least one additional biosynthetic pathway in A. nidulans involving l-arginine or an intermediate metabolite as a substrate. Under certain growth conditions, the addition of l-arginine to liquid media elicited a burst of NO that was not dependent on any of the urea cycle genes. The NO levels were controlled by the metabolically available arginine, which was regulated by mobilization from the vacuoles and during development. In vitro assays with protein extracts and amino acid profiling strongly suggested the existence of an arginine-dependent NO pathway analogous to the mammalian NO synthase. Addition of polyamines induced NO synthesis, and mutations in the polyamine synthesis genes puA and spdA reduced the production of NO. In conclusion, here we report an additional pathway for the synthesis of NO in A. nidulans using urea cycle intermediates.Austrian Federal Ministry of Education, Science and Research P 3279

Nitric oxide synthesis by nitrate reductase is regulated during development in Aspergillus

Nitric oxide (NO) is a signalling molecule involved in many biological processes in bacteria, plants and mammals. However, little is known about the role and biosynthesis of NO in fungi. Here we show that NO production is increased at the early stages of the transition from vegetative growth to development in Aspergillus nidulans. Full NO production requires a functional nitrate reductase (NR) gene (niaD) that is upregulated upon induction of conidiation, even under N-repressing conditions in the presence of ammonium. At this stage, NO homeostasis is achieved by balancing biosynthesis (NR) and catabolism (flavohaemoglobins). niaD and flavohaemoglobin fhbA are transiently upregulated upon induction of conidiation, and both regulators AreA and NirA are necessary for this transcriptional response. The second flavohaemoglobin gene fhbB shows a different expression profile being moderately expressed during the early stages of the transition phase from vegetative growth to conidiation, but it is strongly induced 24 h later. NO levels influence the balance between conidiation and sexual reproduction because artificial strong elevation of NO levels reduced conidiation and induced the formation of cleistothecia. The nitrate-independent and nitrogen metabolite repression-insensitive transcriptional upregulation of niaD during conidiation suggests a novel role for NR in linking metabolism and development.Ministerio de Economía, Industria y Competitividad (MINECO) de España. BIO2012-34381Lower Austria Science Fund. LS12-009Austrian Science Fund (FWF). M01693-B2

El proceso de profesionalización: la evaluación como espacio de construcción de conocimiento

Taking as the starting-point the In-Service Teacher Education School-Based, this paper analyses collaboration as an objetive and strategie for implementing a climate and culture that foster the educational innovation in Primary Education Centres. The authors employ the instruments due to the participant observation to analyse the collaborative reconceptualization and reconstruction of the assessment practices of the pupils from a Colegio Rural Agrupado (CRA)

E2F7 regulates transcription and maturation of multiple microRNAs to restrain cell proliferation

This work was supported by the Spanish Ministry [SAF2012-33551, co-funded by the European RegionalDevelopment fund to A.M.Z., SAF2012-38215 to M.M.,SAF2014-57791-REDC to A.M.Z. and to M.M.]; BasqueGovernment [IT634-13 to A.M.Z.]; University of theBasque Country UPV/EHU [UFI1120 to A.M.Z.]; Excellence Network CellSYS [BFU2014-52125-REDT to M.M.];Comunidad de Madrid [S2010/BMD-2470 to M.M.];Basque Government Fellowship for graduate studies (to J.M.). Funding for open access charge: Basque Government [IT634-13]. Conflict of interest statement. None declared.E2F transcription factors (E2F1-8) are known to coordinately regulate the expression of a plethora of target genes, including those coding for microRNAs (miRNAs), to control cell cycle progression. Recent work has described the atypical E2F factor E2F7 as a transcriptional repressor of cell cycle-related protein-coding genes. However, the contribution of E2F7 to miRNA gene expression during the cell cycle has not been defined. We have performed a genome-wide RNA sequencing analysis to identify E2F7-regulated miRNAs and show that E2F7 plays as a major role in the negative regulation of a set of miRNAs that promote cellular proliferation. We provide mechanistic evidence for an interplay between E2F7 and the canonical E2F factors E2F1-3 in the regulation of multiple miRNAs. We show that miR-25, -26a, -27b, -92a and -7 expression is controlled at the transcriptional level by the antagonistic activity of E2F7 and E2F1-3. By contrast, let-7 miRNA expression is controlled indirectly through a novel E2F/c-MYC/LIN28B axis, whereby E2F7 and E2F1-3 modulate c-MYC and LIN28B levels to impact let-7 miRNA processing and maturation. Taken together, our data uncover a new regulatory network involving transcriptional and post-transcriptional mechanisms controlled by E2F7 to restrain cell cycle progression through repression of proliferation-promoting miRNAs.S