Implications of a Developmental-Stage-Dependent Thylakoid-Bound Protease in the Stabilization of the Light-Harvesting Pigment-Protein Complex Serving Photosystem II during Thylakoid Biogenesis in Red Kidney Bean1

Intact etioplasts of bean (Phaseolus vulgaris) plants exhibit proteolytic activity against the exogenously added apoprotein of the light-harvesting pigment-protein complex serving photosystem II (LHCII) that increases as etiolation is prolonged. The activity increases in the membrane fraction but not in the stroma, where it remains low and constant and is mainly directed against LHCII and protochlorophyllide oxidoreductase. The thylakoid proteolytic activity, which is low in etioplasts of 6-d-old etiolated plants, increases in plants pretreated with a pulse of light or exposed to intermittent-light (ImL) cycles, but decreases during prolonged exposure to continuous light, coincident with chlorophyll (Chl) accumulation. To distinguish between the control of Chl and/or development on proteolytic activity, we used plants exposed to ImL cycles of varying dark-phase durations. In ImL plants exposed to an equal number of ImL cycles with short or long dark intervals (i.e. equal Chl accumulation but different developmental stage) proteolytic activity increased with the duration of the dark phase. In plants exposed to ImL for equal durations to such light-dark cycles (i.e. different Chl accumulation but same developmental stage) the proteolytic activity was similar. These results suggest that the protease, which is free to act under limited Chl accumulation, is dependent on the developmental stage of the chloroplast, and give a clue as to why plants in ImL with short dark intervals contain LHCII, whereas those with long dark intervals possess only photosystem-unit cores and lack LHCII.

HslV-HslU: A novel ATP-dependent protease complex in Escherichia coli related to the eukaryotic proteasome.

We have isolated a new type of ATP-dependent protease from Escherichia coli. It is the product of the heat-shock locus hslVU that encodes two proteins: HslV, a 19-kDa protein similar to proteasome beta subunits, and HslU, a 50-kDa protein related to the ATPase ClpX. In the presence of ATP, the protease hydrolyzes rapidly the fluorogenic peptide Z-Gly-Gly-Leu-AMC and very slowly certain other chymotrypsin substrates. This activity increased 10-fold in E. coli expressing heat-shock proteins constitutively and 100-fold in cells expressing HslV and HslU from a high copy plasmid. Although HslV and HslU could be coimmunoprecipitated from cell extracts of both strains with an anti-HslV antibody, these two components were readily separated by various types of chromatography. ATP stimulated peptidase activity up to 150-fold, whereas other nucleoside triphosphates, a nonhydrolyzable ATP analog, ADP, or AMP had no effect. Peptidase activity was blocked by the anti-HslV antibody and by several types of inhibitors of the eukaryotic proteasome (a threonine protease) but not by inhibitors of other classes of proteases. Unlike eukaryotic proteasomes, the HslVU protease lacked tryptic-like and peptidyl-glutamyl-peptidase activities. Electron micrographs reveal ring-shaped particles similar to en face images of the 20S proteasome or the ClpAP protease. Thus, HslV and HslU appear to form a complex in which ATP hydrolysis by HslU is essential for peptide hydrolysis by the proteasome-like component HslV.

A presynaptic locus for long-term potentiation of elementary synaptic transmission at mossy fiber synapses in culture.

The complex circuitry of the CA3 region and the abundance of collateral connections has made it difficult to study the mossy fiber pathway in hippocampal slices and therefore to establish the site of expression of long-term potentiation at these synapses. Using a novel cell culture system, we have produced long-term potentiation of the elementary synaptic connections on single CA3 pyramidal neurons following tetanic stimulation of individual dentate gyrus granule cells. As is the case for the hippocampal slice, this potentiation was independent of N-methyl-D-aspartate receptor activation, was simulated by application of forskolin, and its induction did not require any modulatory input. The increase in synaptic strength was accompanied by a reduction in the number of failures of transmission and by an increase in the coefficient of variation of the responses and was prevented by presynaptic injection of an inhibitor of protein kinase A. These findings show that mossy fiber long-term potentiation has a presynaptic locus and that its expression is dependent on protein kinase A.

Calcium/calmodulin-dependent kinase II and long-term potentiation enhance synaptic transmission by the same mechanism.

Ca(2+)-sensitive kinases are thought to play a role in long-term potentiation (LTP). To test the involvement of Ca2+/calmodulin-dependent kinase II (CaM-K II), truncated, constitutively active form of this kinase was directly injected into CA1 hippocampal pyramidal cells. Inclusion of CaM-K II in the recording pipette resulted in a gradual increase in the size of excitatory postsynaptic currents (EPSCs). No change in evoked responses occurred when the pipette contained heat-inactivated kinase. The effects of CaM-K II mimicked several features of LTP in that it caused a decreased incidence of synaptic failures, an increase in the size of spontaneous EPSCs, and an increase in the amplitude of responses to iontophoretically applied alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate. To determine whether the CaM-K II-induced enhancement and LTP share a common mechanism, occlusion experiments were carried out. The enhancing action of CaM-K II was greatly diminished by prior induction of LTP. In addition, following the increase in synaptic strength by CaM-K II, tetanic stimulation failed to evoke LTP. These findings indicate that CaM-K II alone is sufficient to augment synaptic strength and that this enhancement shares the same underlying mechanism as the enhancement observed with LTP.

Evidence that the 50-kDa substrate of brefeldin A-dependent ADP-ribosylation binds GTP and is modulated by the G-protein beta gamma subunit complex.

Brefeldin A, a fungal metabolite that inhibits membrane transport, induces the mono(ADP-ribosyl)ation of two cytosolic proteins of 38 and 50 kDa as judged by SDS/PAGE. The 38-kDa substrate has been previously identified as glyceraldehyde-3-phosphate dehydrogenase (GAPDH). We report that the 50-kDa BFA-induced ADP-ribosylated substrate (BARS-50) has native forms of 170 and 130 kDa, as determined by gel filtration of rat brain cytosol, indicating that BARS-50 might exist as a multimeric complex. BARS-50 can bind GTP, as indicated by blot-overlay studies with [alpha-32P]GTP and by photoaffinity labeling with guanosine 5'-[gamma-32P] [beta,gamma-(4-azidoanilido)]triphosphate. Moreover, ADP-ribosylation of BARS-50 was completely inhibited by the beta gamma subunit complex of G proteins, while the ADP-ribosylation of GAPDH was unmodified, indicating that this effect was due to an interaction of the beta gamma complex with BARS-50, rather than with the ADP-ribosylating enzyme. Two-dimensional gel electrophoresis and immunoblot analysis shows that BARS-50 is a group of closely related proteins that appear to be different from all the known GTP-binding proteins.

Sistema especialista para análise pós-evento da proteção em subestações distribuidoras.

É importante que as redes elétricas tenham altos índices de confiabilidade, de forma a se manter a agilidade e a manutenção ideais para um melhor funcionamento. Por outro lado, o crescimento inesperado da carga, falhas em equipamentos e uma parametrização inadequada das funções de proteção tornam a análise de eventos de proteção mais complexas e demoradas. Além disso, a quantidade de informações que pode ser obtida de relés digitais modernos tem crescido constantemente. Para que seja possível uma rápida tomada de decisão e manutenção, esse projeto de pesquisa teve como objetivo a implementação de um sistema completo de diagnóstico que é ativado automaticamente quando um evento de proteção ocorrer. As informações a serem analisadas são obtidas de uma base de dados e de relés de proteção, via protocolo de comunicação IEC 61850 e arquivos de oscilografia. O trabalho aborda o sistema Smart Grid completo incluindo: a aquisição de dados nos relés, detalhando o sistema de comunicação desenvolvido através de um software com um cliente IEC61850 e um servidor OPC e um software com um cliente OPC, que é ativado por eventos configurados para dispará-lo (por exemplo, atuação da proteção); o sistema de pré-tratamento de dados, onde os dados provenientes dos relés e equipamentos de proteção são filtrados, pré-processados e formatados; e o sistema de diagnóstico. Um banco de dados central mantém atualizados os dados de todas essas etapas. O sistema de diagnóstico utiliza algoritmos convencionais e técnicas de inteligência artificial, em particular, um sistema especialista. O sistema especialista foi desenvolvido para lidar com diferentes conjuntos de dados de entrada e com uma possível falta de dados, sempre garantindo a entrega de diagnósticos. Foram realizados testes e simulações para curtos-circuitos (trifásico, dupla-fase, dupla-fase-terra e fase-terra) em alimentadores, transformadores e barras de uma subestação. Esses testes incluíram diferentes estados do sistema de proteção (funcionamento correto e impróprio). O sistema se mostrou totalmente eficaz tanto no caso de disponibilidade completa quanto parcial de informações, sempre fornecendo um diagnóstico do curto-circuito e analisando o funcionamento das funções de proteção da subestação. Dessa forma, possibilita-se uma manutenção muito mais eficiente pelas concessionárias de energia, principalmente no que diz respeito à prevenção de defeitos em equipamentos, rápida resposta a problemas, e necessidade de reparametrização das funções de proteção. O sistema foi instalado com sucesso em uma subestação de distribuição da Companhia Paulista de Força e Luz.

TEXT-MESS: Intelligent, Interactive and Multilingual Text Mining based on Human Language Technologies, TIN2006-15265-C06

The goal of the project is to analyze, experiment, and develop intelligent, interactive and multilingual Text Mining technologies, as a key element of the next generation of search engines, systems with the capacity to find "the need behind the query". This new generation will provide specialized services and interfaces according to the search domain and type of information needed. Moreover, it will integrate textual search (websites) and multimedia search (images, audio, video), it will be able to find and organize information, rather than generating ranked lists of websites.

Proyecto FIRST (Flexible Interactive Reading Support Tool): desarrollo de una herramienta para ayudar a personas con autismo mediante la simplificación de textos

El Trastorno de Espectro Autista (TEA) es un trastorno que impide el correcto desarrollo de funciones cognitivas, habilidades sociales y comunicativas en las personas. Un porcentaje significativo de personas con autismo presentan además dificultades en la comprensión lectora. El proyecto europeo FIRST está orientado a desarrollar una herramienta multilingüe llamada Open Book que utiliza Tecnologías del Lenguaje Humano para identificar obstáculos que dificultan la comprensión lectora de un documento. La herramienta ayuda a cuidadores y personas con autismo transformando documentos escritos a un formato más sencillo mediante la eliminación de dichos obstáculos identificados en el texto. En este artículo se presenta el proyecto FIRST así como la herramienta desarrollada Open Book.

The Virtual Interactive Relationship Between BIM Project Teams: Effective Communication to aid Collaboration in the Design Process

Building Information Modelling (BIM) provides a shared source of information about a built asset, which creates a collaborative virtual environment for project teams. Literature suggests that to collaborate efficiently, the relationship between the project team is based on sympathy, obligation, trust and rapport. Communication increases in importance when working collaboratively but effective communication can only be achieved when the stakeholders are willing to act, react, listen and share information. Case study research and interviews with Architecture, Engineering and Construction (AEC) industry experts suggest that synchronous face-to-face communication is project teams’ preferred method, allowing teams to socialise and build rapport, accelerating the creation of trust between the stakeholders. However, virtual unified communication platforms are a close second-preferred option for communication between the teams. Effective methods for virtual communication in professional practice, such as virtual collaboration environments (CVE), that build trust and achieve similar spontaneous responses as face-to-face communication, are necessary to face the global challenges and can be achieved with the right people, processes and technology. This research paper investigates current industry methods for virtual communication within BIM projects and explores the suitability of avatar interaction in a collaborative virtual environment as an alternative to face-to-face communication to enhance collaboration between design teams’ professional practice on a project. Hence, this paper presents comparisons between the effectiveness of these communication methods within construction design teams with results of further experiments conducted to test recommendations for more efficient methods for virtual communication to add value in the workplace between design teams.

A Global Analysis of of Imperfect Competition in the Banking Systems and the Latest Israeli Regulation Dealing with Market Failures. ACES Working Paper No. 3, 2007

The high concentration of the banking sector is a cross-border phenomenon that has high impact on local and global economies. This paper's main goal is to analyze the factors that impact concentration in the banking systems around the globe. The innovation of this paper is that we combined economic, "economic environment", and culture variables as explanatory variables for this analysis. We found among other things that regulation in the banking system is helpful in order to keep it competitive. We also found that when the society has more individual values rather than collective ones, its banking sector is less concentrated. In the second part of the paper we focused on the Israeli case, showing that although recent indicators of the Israeli banking system indicate a higher level of concentration and lower level of competition, it seems that the recent trend is moving toward less concentration and higher competition.

JAABA: interactive machine learning for automatic annotation of animal behavior

We present a machine learning-based system for automatically computing interpretable, quantitative measures of animal behavior. Through our interactive system, users encode their intuition about behavior by annotating a small set of video frames. These manual labels are converted into classifiers that can automatically annotate behaviors in screen-scale data sets. Our general-purpose system can create a variety of accurate individual and social behavior classifiers for different organisms, including mice and adult and larval Drosophila.

The development of UTCS/NETSIM/ICG: an integrated urban traffic control system - network simulation - interactive computer graphics program. Final report.

Transportation Department, Office of University Research, Washington, D.C.

Interactive simulation of the fate of hazardous chemicals during land treatment of oily wastes : RITZ user's guide /

Mode of access: Internet.