Chloride channels in myotonia congenita assessed by velocity recovery cycles

Introduction: Myotonia congenita (MC) is caused by congenital defects in the muscle chloride channel CLC-1. This study used muscle velocity recovery cycles (MVRCs) to investigate how membrane function is affected. Methods: MVRCs and responses to repetitive stimulation were compared between 18 patients with genetically confirmed MC (13 recessive, 7 dominant) and 30 age-matched normal controls. Results: MC patients exhibited increased early supernormality, but treatment with sodium channel blockers prevented this. After multiple conditioning stimuli, late supernormality was enhanced in all MC patients, indicating delayed repolarization. These abnormalities were similar between the MC subtypes, but recessive patients showed a greater drop in amplitude during repetitive stimulation. Discussion: MVRCs indicate that chloride conductance only becomes important when muscle fibers are depolarized. The differential responses to repetitive stimulation suggest that in dominant MC the affected chloride channels are activated by strong depolarization, consistent with a positive shift of the CLC-1 activation curve. © 2013 Wiley Periodicals, Inc.

Bovine Staphylococcus aureus: Subtyping, evolution, and zoonotic transfer.

Staphylococcus aureus is globally one of the most important pathogens causing contagious mastitis in cattle. Previous studies using ribosomal spacer (RS)-PCR, however, demonstrated in Swiss cows that Staph. aureus isolated from bovine intramammary infections are genetically heterogeneous, with Staph. aureus genotype B (GTB) and GTC being the most prominent genotypes. Furthermore, Staph. aureus GTB was found to be contagious, whereas Staph. aureus GTC and all the remaining genotypes were involved in individual cow disease. In addition to RS-PCR, other methods for subtyping Staph. aureus are known, including spa typing and multilocus sequence typing (MLST). They are based on sequencing the spa and various housekeeping genes, respectively. The aim of the present study was to compare the 3 analytic methods using 456 strains of Staph. aureus isolated from milk of bovine intramammary infections and bulk tanks obtained from 12 European countries. Furthermore, the phylogeny of animal Staph. aureus was inferred and the zoonotic transfer of Staph. aureus between cattle and humans was studied. The analyzed strains could be grouped into 6 genotypic clusters, with CLB, CLC, and CLR being the most prominent ones. Comparing the 3 subtyping methods, RS-PCR showed the highest resolution, followed by spa typing and MLST. We found associations among the methods but in many cases they were unsatisfactory except for CLB and CLC. Cluster CLB was positive for clonal complex (CC)8 in 99% of the cases and typically positive for t2953; it is the cattle-adapted form of CC8. Cluster CLC was always positive for t529 and typically positive for CC705. For CLR and the remaining subtypes, links among the 3 methods were generally poor. Bovine Staph. aureus is highly clonal and a few clones predominate. Animal Staph. aureus always evolve from human strains, such that every human strain may be the ancestor of a novel animal-adapted strain. The zoonotic transfer of IMI- and milk-associated strains of Staph. aureus between cattle and humans seems to be very limited and different hosts are not considered as a source for mutual, spontaneous infections. Spillover events, however, may happen.

Staphylococcus aureus genotype B and other genotypes isolated from cow milk in European countries

Staphylococcus aureus is globally one of the most important pathogens causing contagious mastitis in cattle. Previous studies, however, have demonstrated in Swiss cows that Staph. aureus isolated from bovine intramammary infection is genetically heterogeneous, with Staph. aureus genotype B (GTB) and GTC being the most prominent genotypes. In addition, Staph. aureus GTB was found to be contagious, whereas Staph. aureus GTC and all the remaining genotypes were involved in individual cow disease. The aim of this study was to subtype strains of Staph. aureus isolated from bovine mastitic milk and bulk tank milk to obtain a unified view of the presence of bovine staphylococcal subtypes in 12 European countries. A total of 456 strains of Staph. aureus were subjected to different typing methods: ribosomal spacer PCR, detection of enterotoxin genes, and detection of gene polymorphisms (lukE, coa). Major genotypes with their variants were combined into genotypic clusters (CL). This study revealed 5 major CL representing 76% of all strains and comprised CLB, CLC, CLF, CLI, and CLR. The clusters were characterized by the same genetic properties as the Swiss isolates, demonstrating high clonality of bovine Staph. aureus. Interestingly, CLB was situated in central Europe whereas the other CL were widely disseminated. The remaining 24% of the strains comprised 41 genotypes and variants, some of which (GTAM, GTBG) were restricted to certain countries; many others, however, were observed only once.

Mutation Update of the CLCN5 Gene Responsible for Dent Disease 1

Dent disease is a rare X-linked tubulopathy characterized by low molecular weight proteinuria, hypercalciuria, nephrocalcinosis and/or nephrolithiasis, progressive renal failure, and variable manifestations of other proximal tubule dysfunctions. It often progresses over a few decades to chronic renal insufficiency, and therefore molecular characterization is important to allow appropriate genetic counseling. Two genetic subtypes have been described to date: Dent disease 1 is caused by mutations of the CLCN5 gene, coding for the chloride/proton exchanger ClC-5; and Dent disease 2 by mutations of the OCRL gene, coding for the inositol polyphosphate 5-phosphatase OCRL-1. Herein, we review previously reported mutations (n = 192) and their associated phenotype in 377 male patients with Dent disease 1 and describe phenotype and novel (n = 42) and recurrent mutations (n = 24) in a large cohort of 117 Dent disease 1 patients belonging to 90 families. The novel missense and in-frame mutations described were mapped onto a three-dimensional homology model of the ClC-5 protein. This analysis suggests that these mutations affect the dimerization process, helix stability, or transport. The phenotype of our cohort patients supports and extends the phenotype that has been reported in smaller studies.

Ligand-stimulated internalization of a Dictyostelium discoideum G protein-coupled cAMP receptor

The social amoeba, Dictyostelium discoideum, undergoes a remarkable starvation-induced program of development that transforms a population of unicellular amoebae into a fruiting body composed of resistant spores suspended on a stalk. During this development, secreted cAMP drives chemotaxis of the amoebae, leading to their aggregation, and subsequent differentiation and morphogenesis. Four sequentially expressed G protein-coupled receptors (GPCRs) for cAMP play critical roles in this process. The first of these, cAR1, is essential for aggregation as it mediates chemotaxis as well as the propagation of secreted cAMP waves throughout aggregating populations. Ligand-induced internalization has been shown to regulate a variety of GPCRs. However, little was known at the outset of this study about the role of internalization in the regulation of cAR1 function or, for that matter, in developmental systems in general. For this study, cAMP-induced cAR1 internalization was assessed by measuring (1) the reduction of cell surface binding sites for [ 3H]cAMP and (2) the redistribution of YFP-tagged receptors to the cell's interior, cAMP was found to induce little or no loss of ligand binding (LLB) in vegetative cells. However, the ability to induce LLB increased progressively over the initial 6 hrs of development, reaching ∼70% in cells undergoing aggregation. Despite these reductions in surface binding, detectable cAR1-YFP redistribution could be induced by cAMP only after the cells reached the mound stage (10 hrs) and was found to occur naturally by the ensuing slug stage (18 hrs). Site-directed substitution of a cluster of 5 serines in the receptor's cytoplasmic tail that was previously shown to be the principal site of cAMP-induced cAR1 phosphorylation impaired both LLB and receptor redistribution and furthermore resulted in mound-stage developmental arrest, suggesting that phosphorylation of cAR1 is a prerequisite for its internalization and that cAR1 internalization is required for post-aggregative development. To assess the involvement of clathrin mediated endocytosis, Dictyostelium cells lacking the clathrin light chain gene (clc-) or either of two dynamin genes were examined and found to be defective in LLB and, in the case of clc- cells, also cAR1 redistribution and turnover. Furthermore, cAR1 overexpression in clc- cells (like the serine mutant in wild-type cells) promoted developmental arrest in mounds. The mound-arrest phenotype was also recapitulated in a wild-type background by the specific expression of cAR1 in prestalk cells (but not prespore cells), suggesting that development depends critically on internalization and clearance of cAR1 from these cells. Persistent cAR1 expression following aggregation was found to be associated with aberrant expression of prestalk and prespore genes, which may adversely affect development in the prestalk cell lineage. The PI3 kinase-TORC2 signal transduction pathway, known to be important for Dictyostelium chemotaxis and internalization of yeast pheromone receptors, was examined using chemical inhibitors and null cells and found to be necessary for cAR1 internalization. In conclusion, cAR1 was shown to be similar to other GPCRs in that its internalization depends on phosphorylation of cytoplasmic domain serines, utilizes clathrin and dynamin, and involves the TORC2 complex. In addition, the findings presented here that cAR1 internalization is both developmentally regulated and required for normal development represent a novel regulatory paradigm that might pertain to other GPCRs known to play important roles in the development of humans and other metazoans. ^

Pollen profiles from 8 sediment cores from the Degersee, Southern Germany

The discovery of a neolithic pile field in the shallow water near the eastern shore of the Degersee confirmed earlier palynological and sedimentological studies stating that early man was active in the region since more than 6000 years. The already available off-site data were freshly assessed, completed by additional data from old and new cores and the interpretations revised. A common time scale for the off-site data and the on-site data was obtained by AMS dating of terrestrial macro remains of the neolithic section of off-site core De_I+De_H. The ages can thus be parallelled with AMS ages of construction timber on-site. Pollen analyses from all cores provide a further time scale. The continuously and densely sampled pollen profile of the profundal zone embracing the entire Late glacial and Holocene serves as a reference. From the Boreal onwards the relative ages are transformed by AMS ages and varve counts into calibrated and absolute. A transect cored close to the neolithic pile field across the lake marl-platform demonstrates its geological architecture in the shallow water since the Lateglacial. Studies of the microfabric of thin sections of drilled cores and of box cores from the excavations demonstrate that neolithic settlements now at 2-3,5 m water depth had been erected on lake marl freshly fallen dry, thus indicating earlier lake levels dropped by 1.5-2 m. The neolithic section of the highly resolved off-site profile in the lake=s profundal zone has laminated and calcareous zones alternating with massive ones. Assemblages of diatoms and concentrations of trace elements changing simultaneously characterise the calcareous sections as deposits of low lake levels that lasted between some 40 and more than 300 years. The ages of discovered lake shore dwellings fall into calcareous segments with low lake levels. From the end of the Upper Atlantic period (F VII) appear Secondary Forest Cycles in the beech forest, a man-made sequence of repeated vegetational development with an identical pattern: With a decrease of beech pollen appear pollen of grasses, herbs and cultural indicators. These are suppressed by the light demanding hazel and birch, those again by ash, and finally by the shade demanding beech forming a new pollen peak. Seven main Forest Cycles are identified In the upper Neolithic period each comprising some 250, 450 or 800 years. They are subdivided into subcycles that can be broken down by very dense sampling in even shorter cycles of decadal length. Farming settlers have caused minor patchy clearances of the beech-mixed-forest with the use of fire. The phases of clearance coincide with peaks of charcoal and low stands of the lake levels. The Secondary Forest Cycles and the continuous occurrence of charcoal prove a continued occupation of the region. Together with the repeated restoration of the beech climax forest they point to pulsating occupation probably associated with dynamic demography. The synchronism of the many palynological, sedimentological and archaeological data point to an external forcing as the climate that affects comprehensively all these proxies. The fluctuations of the activity of the sun as manifested in the residual d14C go largely along with the proxies. The initial clearances at the begin of the forest cycles are linked to low lake levels and negative values of d14C that point to dry and warm phases of a more continental climate type. The subcycles exist independent from climatic changes, indicating that early man acted largely independent from external forces.

Pollen record from Blankensee, Germany

Lake Blankensee is filled with 14 m of late- and postglacial deposits, Lake Siethener See with 22,5 m. The lacustrine sedimentation begins in Lake Siethener See in the middle of the Alleröd with annual lamination which partly continues in the Younger Dryas. A 2 cm thick layer of the Laacher See tephra was found in both lakes, the Saksunarvatn tephra only in Lake Siethener See where the cool Rammelbeek-phase (Preboreal) could be shown. The youngest part of the sediment profiles is suspended drifting mud. Masses of Pediastrum (algae) indicate an increasing shoaling of Lake Blankensee after the Subboreal.

Pan-European land cover/use map for 2009

GlobCorine demonstrated an automatic service that can generate in a consistent way land cover / land use maps and land change indicators, based on a CLC-compatible legend. CLC is derived from a visual identification and classification of landscape objects using high resolution images. This methodology provides high thematic accuracy but limits the update rate since it is time-consuming. Therefore, the project evaluated the use of MERIS FR time series, processed automatically to provide a more frequent update of CLC-compatible maps. GlobCorine built upon the experience and resources available through the GlobCover project, to tune the classification chain and adapt it to the EEA needs, covering the pan-European area (including the Mediterranean basin and the European Russia), although the system could be potentially extendable globally. The project delivered two CLC-compatible pan-European land cover maps in less than two years, demonstrating efficient and quick production. The first map is based on Envisat MERIS fine resolution (300m) mode data acquired between end 2004 and mid 2006, while the second used full-year 2009 data. GlobCorine is an initiative of ESA with the partnership of EEA and is implemented by Universite' catholique de Louvain - UCL.

Pollen record from Muggesfelder See and Plussee in Germany

A basaltic tephra layer consisting of brownish-olive glass shards. and about 0.2 mm thick. was found in cores from four lakes in northwest Germany. According to pollen analysis it was deposited during the early Boreal period (corresponding to about 8700 BP). The petrographic properties. the geochemical composition and the age agree with those of the Saksunarvatn tephra. which was first found on the Faroe Islands. The position of the tephra layer in the pollen stratigraphy and in the absolute time-scale is discussed. Procedures for locating the tephra in other cores are suggested.

Análisis de edificación y sostenibilidad

El desarrollo sostenible, es un concepto amplio que afecta a todos los ámbitos de nuestro entorno natural, social y económico. Existen multitud de interrelaciones entre los diferentes ámbitos de éste que nos permitirán realizar un análisis tangible y apoyado en datos sobre las interacciones que se dan. La intención es realizar un análisis de los datos más relevantes de los indicadores del desarrollo sostenible que establecen relaciones con las edificaciones, a nivel del sector de la construcción y del parque residencial y terciario existente en España. No debemos olvidar el análisis de las leyes de la edificación, y de como se ha ido incorporando el desarrollo sostenible en los últimos años y que grado de repercusión ha alcanzado. Para analizar la edificación y sostenibilidad nos debemos respaldar en datos, los cuales se recogen en los indicadores más destacados en materia de desarrollo sostenible como son los: de la EEDS, la UE-EDS, el Banco Público de Indicadores Ambientales (BPIA) y los elaborados por el OSE. El carácter tan singular de la relación que se quiere establecer, provoca la búsqueda de otros muchos datos que establezcan relaciones más íntimas entre las edificaciones o la construcción. Estos datos se han sacado de las fuentes institucionales más relevantes, siempre con la finalidad de dar luz a las relaciones que se establecen entre los edificios y el sector de la construcción con los diferentes aspectos más notables con los que interactúan, en términos de desarrollo sostenible. Las relaciones se han establecido unas veces con la construcción como sector económico, otras atendiendo simplemente a las edificaciones como punto de consumo o desecho (hogares) de ciertos ítems. Las fuentes usadas para la elaboración de este análisis, aparte de los indicadores de sostenibilidad han sido: el Eurostat, proyecto Europeo CORINE Land Cover (CLC), el Instituto Nacional de Estadística (INE), Banco Público de Indicadores Ambientales (BPIA) del MARM, datos de documentos de diversos Ministerios, organismos e instituciones.

The University in the Cloud Computing.

With the advancement of Information and Communication Technology ICT which favors increasingly fast, easy, and accessible communication for all and which can reach large groups of people, there have been changes, in recent years in our society that have modified the way we interact, communicate and transmit information. Access to this, it is possible, not only through computers situated in a fixed location, but new mobile devices make it available, wherever the user happens to be located. Now, information "travels" with the user. These forms of communication, transmission and access to information, have also affected the way to conceive and manage business. To these new forms of business that the Internet has brought, is now added the concept of companies in the Cloud Computing ClC. The ClC technology is based on the supply and consumption of services on demand and pay per use, and it gives a 180 degree turn to the business management concept. Small and large businesses may use the latest developments in ICT, to manage their organizations without the need for expensive investments in them. This will enable enterprises to focus more specifically within the scope of their business, leaving the ICT control to the experts. We believe that education can also and should benefit from these new philosophies. ?Due to the global economic crisis in general and each country in particular, economic cutbacks have come to most universities. These are seen in the need to raise tuition rates, which makes increasingly fewer students have the opportunity to pursue higher education?. In this paper we propose using ClC technologies in universities and we make a dissertation on the advantages that it can provide to both: universities and students. For the universities, we expose two focuses, one: ?to reorganize university ICT structures with the ClC philosophy? and the other one, ?to extend the offer of the university education with education on demand?. Regarding the former we propose to use public or private Clouds, to reuse resources across the education community, to save costs on infrastructure investment, in upgrades and in maintenance of ICT, and paying only for what you use and with the ability to scale according to needs. Regarding the latter, we propose an educational model in the ClC, to increase the current university offerings, using educational units in the form of low-cost services and where students pay only for the units consumed on demand. For the students, they could study at any university in the world (virtually), from anywhere, without travel costs: money and time, and what is most important paying only for what they consume. We think that this proposal of education on demand may represent a great change in the current educational model, because strict registration deadlines disappear, and also the problem of economically disadvantaged students, who will not have to raise large amounts of money for an annual tuition. Also it will decrease the problem of loss of the money invested in an enrollment when the student dropout. In summary we think that this proposal is interesting for both, universities and students, we aim for "Higher education from anywhere, with access from any mobile device, at any time, without requiring large investments for students, and with reuse and optimization of resources by universities. Cost by consumption and consumption by service?. We argue for a Universal University "wisdom and knowledge accessible to all?

Same Virtus 120, versatilidad y economía sin complicaciones

El pasado día 17 de junio de 2014 tuvimos la oportunidad de realizar ensayos en campo a un tractor Same Virtus 120, en las instalaciones que la empresa posee junto a Toledo. La prueba de laboreo se realizó con un cultivador Kverneland CLC Evo con nueve brazos amortiguados con ballestas, rejas rectas y 2,4 m de anchura.También realizamos una prueba de transporte de un remolque con cebada, y evaluamos la maniobrabilidad y frenada del tractor.

Análisis energético de las centrales de ciclo combinado con gasificación integrada y combustión mediante lazo químico

La combustión mediante lazo químico permite una integración de captura de CO2 en una planta térmica de potencia sin penalización energética. Se alcanza una menor destrucción exergética, alcanzándose un rendimiento térmico idóneo. Este proyecto se centra en el estudio del rendimiento energético en una planta de potencia con gasificación integrada cuando la cámara de combustión de las turbinas de gas se sustituye por un sistema de combustión CLC. Después del diseño termodinámico y optimización de algunos parámetros del ciclo, el rendimiento de la planta de potencia es evaluado bajo distintas condiciones de trabajo y comparado con el que se obtendría en una planta de ciclo combinado convencional con gasificación integrada y captura pre-combustión. Abstract Chemical-looping combustion allows an integration of CO2 capture in a thermal power plant without energy penalty. A less exergy destruction in the combustion chemical transformation is achieved, leading to a greater overall thermal efficiency. This Project focus on the study of the energetic performance in a cycle power plant with integrated gasification when the chamber combustion is supplied by a combustion system CLC. After thermodynamic modeling and optimization of some cycle parameters, the power plant performance is evaluated under diverse working conditions and compared to a conventional integrated gasification combined cycle with pre-combustion capture.

In Ecclesiasten Salomonis annotationes piae et eruditae : ex variis cum veterum orthodoxorum, tum recentiorum scriptis congestae, ac olim etiam pro concione enarratae, anno MDXXXIIII Moguntiae /

CCBE S. XVI, W, 107-108 ; Adams F353 ; CLC W366