A fluorometric method for the assay of protein kinase activity

Protein kinases constitute one of the largest protein families in nature. Current methods to assay their activity involve the use of radioactive ATP or very expensive reagents. In this work, we developed a highly sensitive, cost-effective and straightforward protocol to measure protein kinase activity using a microplate layout. Released ADP is converted into NAD+, which is quantified by its fluorescent properties after alkaline treatment (linear range 0–10 nmol ADP). To validate our protocol, we characterized a recombinant calcium-dependent protein kinase from potato. Overall, this tool represents a critical step forward in the functional characterization of protein kinases.Fil: Rojas, Bruno Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; ArgentinaFil: Santin, Franco. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Ulloa, Rita Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Iglesias, Alberto Alvaro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; ArgentinaFil: Figueroa, Carlos Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentin

Using semantics for automating the authentication of Web APIs

Recent technology developments in the area of services on the Web are marked by the proliferation of Web applications and APIs. The implementation and evolution of applications based on Web APIs is, however, hampered by the lack of automation that can be achieved with current technologies. Research on semantic Web services is there fore trying to adapt the principles and technologies that were devised for traditional Web services, to deal with this new kind of services. In this paper we show that currently more than 80% of the Web APIs require some form of authentication. Therefore authentication plays a major role for Web API invocation and should not be neglected in the context of mashups and composite data applications. We present a thorough analysis carried out over a body of publicly available APIs that determines the most commonly used authentication approaches. In the light of these results, we propose an ontology for the semantic annotation of Web API authentication information and demonstrate how it can be used to create semantic Web API descriptions. We evaluate the applicability of our approach by providing a prototypical implementation, which uses authentication annotations as the basis for automated service invocation

Architecture and origin of fluvial cross-bedding based on flume experiments and geological examples field case studies: Rillo de Gallo, Spain and Northumberland, UK

Cross-stratified sandstones formed by dunes and unit bars is common in the rock record. Cross-bed architecture is controlled by the size and shape of, and processes on, the original formative bedforms, and processes that truncate, deform or bury the cross-strata. This thesis reports an investigation of cross-bedded sandstones and their internal sedimentary structures formed in fluvial environments within different depositional settings at Rillo de Gallo, (Spain) and Seaton Sluice, (Northumberland). In addition, flume experiments using well-sorted fine sand were used to investigate some of the factors controlling cross-bed architecture produced by unit bars migrating downstream in a unidirectional flow under controlled conditions over pre-existing topography. New ways of classifying cross-bedding were developed to allow analyses of patterns and relationships between set geometry, flow behaviour and stratigraphic position within sedimentary sequences. The amount and variation in divergence between scour trend and scour-fill laminae dip direction is found to be potentially diagnostic of flow pattern, and should improve palaeoenvironment interpretation. A relationship between the amount of variation in bar lee-face angle with bar height was found in the flume and may be useful for inferring bedform size in the rock record. Integration of laboratory and field observations on lee-face angle, bedform superimposition, reactivation surfaces, topography development and bottomset formation helped to mitigate the issue of preservation level when interpreting rock analogue examples and led to improved interpretations of the ancient fluvial deposits. This is an approach that should help the interpretation of other ancient fluvial sequences. The mechanisms and patterns of trough scour formation and scour filling have been to a certain extent ignored to date. Herein it is highlighted the need for further research on the understanding of scour formation and fill in association with bedform development and migration leading to improved knowledge of cross-bedded sandstones

Cholangiocytes: Cell transplantation

Background:Due to significant limitations to the access to orthotropic liver transplantation, cell therapies forliver diseases have gained large interest worldwide.Scope of review:To revise current literature dealing with cell therapy for liver diseases. We discussed the ad-vantages and pitfalls of the different cell sources tested so far in clinical trials and the rationale underlying thepotential benefits of transplantation of human biliary tree stem cells (hBTSCs).Major conclusions:Transplantation of adult hepatocytes showed transient benefits but requires immune-sup-pression that is a major pitfall in patients with advanced liver diseases. Mesenchymal stem cells and hemato-poietic stem cells transplanted into patients with liver diseases are not able to replace resident hepatocytes butrather they target autoimmune or inflammatory processes into the liver. Stem cells isolated from fetal or adultliver have been recently proposed as alternative cell sources for advanced liver cirrhosis and metabolic liverdisease. We demonstrated the presence of multipotent cells expressing a variety of endodermal stem cell markersin (peri)-biliary glands of bile ducts in fetal or adult human tissues, and in crypts of gallbladder epithelium. Inthefirst cirrhotic patients treated in our center with biliary tree stem cell therapy, we registered no adverse eventbut significant benefits.General significance:The biliary tree stem cell could represent the ideal cell source for the cell therapy of liverdiseases. This article is part of a Special Issue entitled: Cholangiocytes in Health and Diseaseedited by JesusBanales, Marco Marzioni, Nicholas LaRusso and Peter Jansen

NILM techniques for intelligent home energy management and ambient assisted living: a review

The ongoing deployment of smart meters and different commercial devices has made electricity disaggregation feasible in buildings and households, based on a single measure of the current and, sometimes, of the voltage. Energy disaggregation is intended to separate the total power consumption into specific appliance loads, which can be achieved by applying Non-Intrusive Load Monitoring (NILM) techniques with a minimum invasion of privacy. NILM techniques are becoming more and more widespread in recent years, as a consequence of the interest companies and consumers have in efficient energy consumption and management. This work presents a detailed review of NILM methods, focusing particularly on recent proposals and their applications, particularly in the areas of Home Energy Management Systems (HEMS) and Ambient Assisted Living (AAL), where the ability to determine the on/off status of certain devices can provide key information for making further decisions. As well as complementing previous reviews on the NILM field and providing a discussion of the applications of NILM in HEMS and AAL, this paper provides guidelines for future research in these topics.Agência financiadora: Programa Operacional Portugal 2020 and Programa Operacional Regional do Algarve 01/SAICT/2018/39578 Fundação para a Ciência e Tecnologia through IDMEC, under LAETA: SFRH/BSAB/142998/2018 SFRH/BSAB/142997/2018 UID/EMS/50022/2019 Junta de Comunidades de Castilla-La-Mancha, Spain: SBPLY/17/180501/000392 Spanish Ministry of Economy, Industry and Competitiveness (SOC-PLC project): TEC2015-64835-C3-2-R MINECO/FEDERinfo:eu-repo/semantics/publishedVersio

Productive local condition and requirements for the commercialization: Transformation in the fruit growing of the north of Argentine Patagonia

A partir de la década de los noventa, en un marco de reestructuración del sistema agroalimentario mundial y desregulación de la política agropecuaria en la Argentina, la producción primaria de alimentos se encuentra sujeta a criterios de calidad internacionales, y regida por requerimientos formales que modifican sustancialmente las condiciones productivas. El caso en estudio aborda las transformaciones socio–productivas en la cadena frutícola localizada en el norte de la Patagonia Argentina, donde estas exigencias, entre otros procedimientos, se formalizan en las Buenas Prácticas Agrícolas. Las mismas son abordadas en tanto dispositivos de control que interpelan y complejizan las condiciones de reproducción social de los chacareros como productores independientes en la cadena, con impactos en sus trayectorias vitales y productivas. Trabajamos con fuentes secundarias y relevamiento primario de casos. Las conclusiones se inscriben en las tensiones entre configuraciones productivas locales y avances de los controles del capital concentrado.From the nineties’ decade, in a context of restructuring of the food–processing world system and deregulation of the agricultural politics in the Argentina, the primary production of food is tied to international criteria of quality, and governed by formal requirements that modify substantially the productive conditions. We study the case of the productive transformations in the fresh fruit chain located in the north of the Patagonia Argentina, where these requirements are formalized in the Good Agricultural Practices. These requirements are approached as devices of control that make more complex than before the conditions of social reproduction of the farmers, independent producers in the chain. We work with secondary sources and primary report of cases. The conclusions register in the tensions between productive local configurations and advances of the controls of the concentrated capital.Fil: Trpin, Verónica. Universidad Nacional del Comahue. Facultad de Derecho y Ciencias Sociales. Grupo de Estudios Sociales Agrarios; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Alvaro, Maria Belen. Universidad Nacional del Comahue. Facultad de Derecho y Ciencias Sociales. Grupo de Estudios Sociales Agrarios; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

VEGF-Mediated Signal Transduction in Tumor Angiogenesis

The vascular endothelial growth factor-A (VEGF) plays a crucial role in tumor angiogenesis. Through its primary receptor VEGFR-2, VEGF exerts the activity of a multitasking cytokine, which is able to stimulate endothelial cell survival, invasion and migration into surrounding tissues, proliferation, as well as vascular permeability and inflammation. The core components of VEGF signaling delineate well-defined intracellular routes. However, the whole scenario is complicated by the fact that cascades of signals converge and branch at many points in VEGF signaling, thus depicting a complex signal transduction network that is also finely regulated by different mechanisms. In this chapter, we present a careful collection of the best-characterized VEGF-induced signal transduction pathways, attempting to offer an overview of the complexity of VEGF signaling in the context of tumor angiogenesis

Terrain Model Resolution Effect on Sight Distance on Roads

Available sight distance is a key factor on road design. This research is focused on the effect of Digital Terrain Model (DTM) resolution on available sight distance. In addition, the effect of distance between calculation points along the vehicle path is also considered. The available sight distance is calculated using a Geographic Information System (GIS). Three high resolution DTMs, based on airborne LiDAR (Light Detection and Ranging), are used. Vehicle path is collected by a Global Navigation Satellite System (GNSS) mounted in a vehicle travelling along the road. Twelve combinations of DTM resolution and distance between calculation points along the vehicle path are studied. Results are statistically analysed and each case study is compared to a reference case which is considered close to reality. The analysis made shows that DTM resolution has a larger effect on the quality of the results than distance between calculation points. Practical issues are discussed

Mapping and Functional Role of Phosphorylation Sites in the Thyroid Transcription Factor-1 (TTF-1)

The phosphorylation of thyroid transcription factor-1 (TTF-1), a homeodomain-containing transcription factor that is required for thyroid-specific expression of the thyroglobulin and thyroperoxidase gene promoters, has been studied. Phosphorylation occurs on a maximum of seven serine residues that are distributed in three tryptic peptides. Mutant derivatives of TTF-1, with alanine residues replacing the serines in the phosphorylation sites, have been constructed and used to assess the functional relevance of TTF-1 phosphorylation. The DNA binding activity of TTF-1 appears to be phosphorylation-independent, as indicated also by the performance of TTF-1 purified from an overexpressing Escherichia coli strain. Transcriptional activation by TTF-1 could require phosphorylation only in specific cell types since in a co-transfection assay in heterologous cells both wild-type and mutant proteins show a similar transcriptional activity