Generation of thermo-sensitive allele of the TPR like protein Nup211by PCR mutagenesis

Motivation: TPR proteins are conserved large coiled-coil proteins that localize at the nucleoplasmic side of the nuclear pore complex and participate in multiple aspects of DNA metabolism. The protein Nup211, fission yeast homolog of Mlp1/Mlp2/Tpr, participate in the mRNA export and is essential for vegetative growth. The aim of this work is to create a collection of thermo-sensitive alleles of nup211.Methods: To create the collection, we have generated a new strain with the nup211 gen tagged with GFP at the amino terminal extreme and confirmed by fluorescent microscopy that the protein Nup211 localized in the nuclear envelop. Then, we have carried out a Taq PCR-based Random Mutagenesis with reduced concentration of dATP. The PCR products were transformed into a wild type strain to generate conditional mutants. The transformants obtained whose growth was impaired at 36ºC were preselected as thermo-sensitive mutants. To confirm the growth deficiency of these clones, a drop assay was performed and the best candidates were selected. These thermo-sensitive mutants were cultivated at 25ºC as well as 36ºC and both cultures were subjected to various experiments in order to study any changes in the localization of Nup211.Results: Up to now, we have demonstrated by fluorescent microscopy that the thermo-sensitive mutants show a modified nuclear distribution of Nup211 and different cellular phenotype, suggesting that the differents clones might represent differents nup211 thermo-sensitive alleles. These alleles are going to be subjected to various experiments to clarify the role of the protein in the mRNA export

Cation distribution of cobalt ferrite electrosynthesized nanoparticles: A methodological comparison

Final publication at http://doi.org/10.1016/j.jallcom.2017.12.342, © 2017 Elsevier B.V.The present work seeks to analyse the structural and magnetic properties of cobalt ferrite nanoparticles obtained by electrochemical synthesis by high-resolution transmission electronic microscopy (HRTEM), X-ray absorption spectroscopy (XAS), Mössbauer spectroscopy (MS), neutron diffraction (ND) and SQUID magnetometer. The cationic distribution is analyzed by different techniques. The inversion degree determined by the most accurate measurements was 0.73(1), and the formula for the nanoparticles therefore was (↑Co 0.27 Fe 0.73 )[↓Co 0.73 Fe 1.27 ]O 4 . The magnetic moment found from DC and Mössbauer spectroscopy measurements was 3.8(3) μB, and the coercivity was 7870 Oe at 100 K.This work is supported by the MINECO/FEDER Project MAT2015-67557-C2-2-

Flow cytometric cell cycle analysis of cultured brown bear fibroblast cells

The aim of this study was to assess by flow cytometry the cell cycle of brown bear fibroblast cells cultured under different growth conditions. Skin biopsies were taken in Cantabria (Spain) from a live, anaesthetized brown bear. DNA analysis was performed by flow cytometry following cell DNA staining with propidium iodide. Serum starvation increased (P < 0.01) the percentage of G0/G1 phase cells (92.7 0.86) as compared to cycling cells (39.7 0.86) or cells cultured to confluency (87.3 0.86). DMSO included for 48 h in the culture significantly increased (P < 0.01) the percentage of G0/G1 phase of the cell cycle at all concentrations used and decreased percentages of S phase in a dose-dependent fashion. Roscovitine increased the G0/G1 phase of the cell cycle (P < 0.01) at 15 mM concentration. Interestingly, the G2/M stage significantly increased at 30 and 50 mM compared to the control and 15 mM (P < 0.02). The cell cycle of brown bear adult fibroblast cells can be successfully synchronized under a variety of culture conditions

PtRu nanoparticles supported on noble carbons for ethanol electrooxidation

In this work, three cytosine derived nitrogen doped carbonaceous materials (noble carbons, NCs) with different atomic C/N ratios and porous networks have been synthesized and used as supports for PtRu electrocatalysts in the ethanol oxidation reaction (EOR) for clean hydrogen production. Both, the metal phase and the carbon support play critical roles in the electrocatalysts final performance. Lower NPs size distribution was obtained over supports with low atomic C/N ratios (i.e., 4 and 6) and defined porosity (i.e., 1701 m2 g−1 for PtRu/CNZ and 1834 m2 g−1 for PtRu/CLZ, respectively). In contrast, a lower C/N ratio and poor porous network (i.e., 65 m2 g−1, PtRu/CLK) led to the largest particle size and fostered an increase of the alloying degree between Pt and Ru NPs (i.e., 3 for C/N ~ 6 and 28 for C/N ~ 3). Electrochemical active surface area was found to increase with decreasing NPs size and the alloy extent, due to a higher availability of Pt active sites. Accelerated degradation tests showed that PtRu/NCs outperform similar to PtRu NPs on commercial carbon pointing at the stabilizing effect of NCs. PtRu/CNZ exhibited the best electrochemical performance (i.e., 69.1 mA mgPt−1), outperforming PtRu/CLZ and PtRu/CLK by 3- and 9-fold, respectively, due to a suitable compromise between particle sizes, degree of alloy, textural properties and elemental composition. Best anodes were scaled-up to a proton exchange membrane cell and PtRu/CNZ was proved to provide the best electrocatalytic activity (262 mA cm−2 and low energy requirements), matching the values obtained by the state of the art of EOR electrocatalysts

Preliminary analysis of the diversity of Chironomidae (Diptera) present in water courses in Catamarca

The diversity of Chironomidae larvae was studied in two water courses in Catamarca, the Río River and El Simbolar Stream. They are both located in ecoregions, the former in the Puna and the latter in the Chaco Serrano. The aim of this paper was to analyze the diversity of Chironomidae in relation to environmental variables in watercourses in Catamarca. Sampling was performed in periods of high and low water, using a Surber sampler (300 μ and 0.09 m-2).Instituto de Limnología "Dr. Raúl A. Ringuelet

Semi-automated morphological characterization using South Rivers Toolbox

Automatic morphological characterization of river systems is important because it provides valuable information on river behavior, helps quantify fluvial changes, improves model accuracy, and supports the management and restoration of river systems. In recent years, scientific interest in the development and use of automated tools for the geomorphological characterization of rivers has increased. The objective of this study was to characterize in a semi-automated way the River Styles of the Duqueco River in south-central Chile. To achieve this objective, an experimental complement within QGIS called South Rivers Toolbox (SRT) was developed. This toolbox allows users to characterize and classify River Styles based on hydromorphological metrics extracted at segment scale, implementing a semi-automatic approach that no longer relies on subjective expert judgment. As a result of the analysis, 43 Rivers Styles have been classified and characterized using the SRT. The upper zone is characterized by being mostly confined, single-channel, with a bed dominated by boulders and blocks, this section of the river is highly affected by hydropower generation plants; the middle zone is confined and single-channel, with a change in the granulometry more dominated by boulders. The lower zone is more diverse in styles, unconfined, with a high presence of geomorphic units that give way to multichannel styles and transition zones. Using the SRT program it is possible to retrospectively analyze the evolution of the river, identifying sections sensitive to geomorphological modification, which is very useful to synoptically evaluate the flood risk, given that, traditionally, for this type of studies only hydraulic modeling is implemented at a river section scale, often with total lack of knowledge of how the river functions and evolves.</p

Multiple Local and Recent Founder Effects of TGM1 in Spanish Families

<div><h3>Background</h3><p>Mutations in the <em>TGM1</em> gene encoding transglutaminase 1 are a major cause of autosomal recessive congenital ichthyosis. In the Galician (NW Spain) population, three mutations, c.2278C>T, c.1223_1227delACAC and c.984+1G>A, were observed at high frequency, representing ∼46%, ∼21% and ∼13% of all <em>TGM1</em> gene mutations, respectively. Moreover, these mutations were reported only once outside of Galicia, pointing to the existence of historical episodes of local severe genetic drift in this region.</p> <h3>Methodology/Principal Findings</h3><p>In order to determine whether these mutations were inherited from a common ancestor in the Galician population, and to estimate the number of generations since their initial appearance, we carried out a haplotype-based analysis by way of genotyping 21 SNPs within and flanking the <em>TGM1</em> gene and 10 flanking polymorphic microsatellite markers spanning a region of 12 Mb. Two linkage disequilibrium based methods were used to estimate the time to the most recent common ancestor (TMRCA), while a Bayesian-based procedure was used to estimate the age of the two mutations. Haplotype reconstruction from unphased genotypes of all members of the affected pedigrees indicated that all carriers for each of the two mutations harbored the same haplotypes, indicating common ancestry.</p> <h3>Conclusions/Significance</h3><p>In good agreement with the documentation record and the census, both mutations arose between 2,800–2,900 years ago (y.a.), but their TMRCA was in the range 600–1,290 y.a., pointing to the existence of historical bottlenecks in the region followed by population growth. This demographic scenario finds further support on a Bayesian Coalescent Analysis based on <em>TGM1</em> haplotypes that allowed estimating the occurrence of a dramatic reduction of effective population size around 900–4,500 y.a. (95% highest posterior density) followed by exponential growth.</p> </div

From chemosynthesis-based communities to cold-water corals: Vulnerable deep-sea habitats of the Gulf of Cádiz

The Gulf of Cádiz (GoC) represents an area of ecological importance within the northeastern Atlantic Ocean due to the presence of Mediterranean and Atlantic water masses, a heterogeneous seafloor and a biological confluence. Nevertheless, information on the presence of vulnerable deep-sea habitats is still very scarce and it is of importance for further habitat monitoring within the context of the Habitats and Marine Strategy Framework Directives and for improving conservation and resource extraction management. From 2010 to 2012, fluid migration and emission related edifices (e.g., mud volcanoes, diapirs) from the Spanish continental margin of the GoC have been explored using a remotely operated vehicle (ROV; Liropus 2000) and an underwater camera sled (UCS; APHIA 2012) as well as several devices for collecting sediment and fauna. Different vulnerable deep-sea habitats have been observed, including anoxic bottoms with bacterial mats, sea-pen communities, sponge aggregations, antipatharian and gorgonian communities and also cold-water coral banks. Some of these habitats are included in conservation lists of the habitat directive and in international conventions (OSPAR, RAC/SPA), however some of them are located in areas of the GoC that are exposed to intense trawling. The diversity of habitats detected in the Spanish continental margin of the GoC highlights the importance of seepage related edifices as inducers of seabed and habitat heterogeneity in deep-sea areas.En prens