The Xanthomonas citri effector protein PthA interacts with citrus proteins involved in nuclear transport, protein folding and ubiquitination associated with DNA repair

P>Xanthomonas axonopodis pv. citri utilizes the type III effector protein PthA to modulate host transcription to promote citrus canker. PthA proteins belong to the AvrBs3/PthA family and carry a domain comprising tandem repeats of 34 amino acids that mediates protein-protein and protein-DNA interactions. We show here that variants of PthAs from a single bacterial strain localize to the nucleus of plant cells and form homo- and heterodimers through the association of their repeat regions. We hypothesize that the PthA variants might also interact with distinct host targets. Here, in addition to the interaction with alpha-importin, known to mediate the nuclear import of AvrBs3, we describe new interactions of PthAs with citrus proteins involved in protein folding and K63-linked ubiquitination. PthAs 2 and 3 preferentially interact with a citrus cyclophilin (Cyp) and with TDX, a tetratricopeptide domain-containing thioredoxin. In addition, PthAs 2 and 3, but not 1 and 4, interact with the ubiquitin-conjugating enzyme complex formed by Ubc13 and ubiquitin-conjugating enzyme variant (Uev), required for K63-linked ubiquitination and DNA repair. We show that Cyp, TDX and Uev interact with each other, and that Cyp and Uev localize to the nucleus of plant cells. Furthermore, the citrus Ubc13 and Uev proteins complement the DNA repair phenotype of the yeast Delta ubc13 and Delta mms2/uev1a mutants, strongly indicating that they are also involved in K63-linked ubiquitination and DNA repair. Notably, PthA 2 affects the growth of yeast cells in the presence of a DNA damage agent, suggesting that it inhibits K63-linked ubiquitination required for DNA repair.

Overexpression of Nrp/b (nuclear restrict protein in brain) suppresses the malignant phenotype in the C6/ST1 glioma cell line

Upon searching for glucocorticoid-regulated cDNA sequences associated with the transformed to normal phenotypic reversion of C6/ST1 rat glioma cells, we identified Nrp/b (nuclear restrict protein in brain) as a novel rat gene. Here we report on the identification and functional characterization of the complete sequence encoding the rat NRP/B protein. The cloned cDNA presented a 1767 nucleotides open-reading frame encoding a 589 aminoacids residues sequence containing a BTB/POZ (broad complex Tramtrack bric-a-brac/Pox virus and zinc finger) domain in its N-terminal region and kelch motifs in its C-terminal region. Sequence analysis indicates that the rat Nrp/b displays a high level of identity with the equivalent gene orthologs from other organisms. Among rat tissues, Nrp/b expression is more pronounced in brain tissue. We show that overexpression of the Nrp/b cDNA in C6/ST1 cells suppresses anchorage independence in vitro and tumorigenicity in vivo, altering their malignant nature towards a more benign phenotype. Therefore, Nrp/b may be postulated as a novel tumor suppressorgene, with possible relevance for glioblastoma therapy. (C) 2009 Elsevier Ltd. All rights reserved.

Sdo1p, the yeast orthologue of Shwachman-Bodian-Diamond syndrome protein, binds RNA and interacts with nuclear rRNA-processing factors

The Shwachman-Bodian-Diamond syndrome protein (SBDS) is a member of a highly conserved protein family of not well understood function, with putative orthologues found in different organisms ranging from Archaea, yeast and plants to vertebrate animals. The yeast orthologue of SBDS, Sdo1p, has been previously identified in association with the 60S ribosomal subunit and is proposed to participate in ribosomal recycling. Here we show that Sdo1p interacts with nucleolar rRNA processing factors and ribosomal proteins, indicating that it might bind the pre-60S complex and remain associated with it during processing and transport to the cytoplasm. Corroborating the protein interaction data, Sdo1p localizes to the nucleus and cytoplasm and co-immunoprecipitates precursors of 60S and 40S subunits, as well as the mature rRNAs. Sdo1p binds RNA directly, suggesting that it may associate with the ribosomal subunits also through RNA interaction. Copyright (C) 2009 John Wiley & Sons, Ltd.

Stoichiometry and thermodynamics of the interaction between the C-terminus of human 90 kDa heat shock protein Hsp90 and the mitochondrial translocase of outer membrane Tom70

A large majority of the 1000-1500 proteins in the mitochondria are encoded by the nuclear genome, and therefore, they are translated in the cytosol in the form and contain signals to enable the import of proteins into the organelle. The TOM complex is the major translocase of the outer membrane responsible for preprotein translocation. It consists of a general import pore complex and two membrane import receptors, Tom20 and Tom70. Tom70 contains a characteristic TPR domain, which is a docking site for the Hsp70 and Hsp90 chaperones. These chaperones are involved in protecting cytosolic preproteins from aggregation and then in delivering them to the TOM complex. Although highly significant, many aspects of the interaction between Tom70 and Hsp90 are still uncertain. Thus, we used biophysical tools to study the interaction between the C-terminal domain of Hsp90 (C-Hsp90), which contains the EEVD motif that binds to TPR domains, and the cytosolic fragment of Tom70. The results indicate a stoichiometry of binding of one monomer of Tom70 per dimer of C-Hsp90 with a K(D) of 360 30 nM, and the stoichiometry and thermodynamic parameters obtained suggested that Tom70 presents a different mechanism of interaction with Hsp90 when compared with other TPR proteins investigated. (C) 2011 Elsevier Inc. All rights reserved.

Nuclear magnetic resonance characterization of metabolite disorder in orange trees caused by citrus sudden death disease

Citrus sudden death (CSD) is a new disease of sweet orange and mandarin trees grafted on Rangpur lime and Citrus volkameriana rootstocks. It was first seen in Brazil in 1999, and has since been detected in more than four million trees. The CSD causal agent is unknown and the current hypothesis involves a virus similar to Citrus tristeza virus or a new virus named Citrus sudden death-associated virus. CSD symptoms include generalized foliar discoloration, defoliation and root death, and, in most cases, it can cause tree death. One of the unique characteristics of CSD disease is the presence of a yellow stain in the rootstock bark near the bud union. This region also undergoes profound anatomical changes. In this study, we analyse the metabolic disorder caused by CSD in the bark of sweet orange grafted on Rangpur lime by nuclear magnetic resonance (NMR) spectroscopy and imaging. The imaging results show the presence of a large amount of non-functional phloem in the rootstock bark of affected plants. The spectroscopic analysis shows a high content of triacylglyceride and sucrose, which may be related to phloem blockage close to the bud union. We also propose that, without knowing the causal CSD agent, the determination of oil content in rootstock bark by low-resolution NMR can be used as a complementary method for CSD diagnosis, screening about 300 samples per hour.

Nuclear magnetic resonance water relaxation time changes in bananas during ripening: a new mechanism

BACKGROUND: Nuclear magnetic resonance studies of banana fragments during ripening show an increase on the water transverse relaxation time (T(2)) and a decrease in water self-diffusion coefficient (D). As T(2) and D are normally directly correlated, we studied these two properties in intact bananas during ripening, in an attempt to rule out the effect of injury on the apparent discrepancies in the behavior of T(2) and D. RESULTS: The results show that injury in bananas causes a decrease in T(2) of the water in vacuoles (T(2vac)). They also show that T(2vac) increased and D decreased during ripening, ruling out the injury effect. To explain the apparent discrepancies, we propose a new hypothesis for the increase in T(2) values, based on the reduction of Fe(3+) ions to Fe(2+) by galacturonic acid, produced by the hydrolysis of pectin and a decrease in internal oxygen concentration during ripening. CONCLUSION: As injury alters T(2) values it is necessary to use intact bananas to study relaxation times during ripening. The novel interpretation for the increase in T(2vac) based on reduction of Fe(+3) and O(2) concentration is an alternative mechanism to that based on the hydrolysis of starch in amyloplasts. (C) 2010 Society of Chemical Industry

Fast and Simple Nuclear Magnetic Resonance Method To Measure Conjugated Linoleic Acid in Beef

Conjugated linoleic acids (CLAs) are a group of linoleic acid isomers that are naturally found in food products originating from ruminants (meat and dairy). These acids have received special attention in recent years due to their potential human health benefits. Research efforts have been proposed to increase the CLA content in beef to improve public health. However, because there are more than 30 million beef cattle used each year by the American food industry, it will be necessary to ensure their content in a large number of samples. Therefore, it is important to have an inexpensive and rapid analytical method to measure CLA content in food products. Because gas chromatography (GC), a current popular method for measuring CLAs, is slow, this paper describes a nuclear magnetic resonance spectroscopy ((1)H NMR) method that is potentially >10 times faster than the GC method. Analyses show a correlation coefficient of 0.97, indicating the capacity of NMR to quantify the CLA content in beef samples. Furthermore, the method proposed herein is simple and does not require sophisticated sample preparation.

Road and Traffic Signs Recognition using Vector Machines

Intelligent Transportation System (ITS) is a system that builds a safe, effective and integrated transportation environment based on advanced technologies. Road signs detection and recognition is an important part of ITS, which offer ways to collect the real time traffic data for processing at a central facility.This project is to implement a road sign recognition model based on AI and image analysis technologies, which applies a machine learning method, Support Vector Machines, to recognize road signs. We focus on recognizing seven categories of road sign shapes and five categories of speed limit signs. Two kinds of features, binary image and Zernike moments, are used for representing the data to the SVM for training and test. We compared and analyzed the performances of SVM recognition model using different features and different kernels. Moreover, the performances using different recognition models, SVM and Fuzzy ARTMAP, are observed.

A Unified Load Generator for Geographically Distributed Generation ofNetwork Traffic

During the last decade, the Internet usage has been growing at an enormous rate which has beenaccompanied by the developments of network applications (e.g., video conference, audio/videostreaming, E-learning, E-Commerce and real-time applications) and allows several types ofinformation including data, voice, picture and media streaming. While end-users are demandingvery high quality of service (QoS) from their service providers, network undergoes a complex trafficwhich leads the transmission bottlenecks. Considerable effort has been made to study thecharacteristics and the behavior of the Internet. Simulation modeling of computer networkcongestion is a profitable and effective technique which fulfills the requirements to evaluate theperformance and QoS of networks. To simulate a single congested link, simulation is run with asingle load generator while for a larger simulation with complex traffic, where the nodes are spreadacross different geographical locations generating distributed artificial loads is indispensable. Onesolution is to elaborate a load generation system based on master/slave architecture.

A Contour-based Separation of VerticallyAttached Traffic Signs

This report presents an algorithm for locating the cut points for and separatingvertically attached traffic signs in Sweden. This algorithm provides severaladvanced digital image processing features: binary image which representsvisual object and its complex rectangle background with number one and zerorespectively, improved cross correlation which shows the similarity of 2Dobjects and filters traffic sign candidates, simplified shape decompositionwhich smoothes contour of visual object iteratively in order to reduce whitenoises, flipping point detection which locates black noises candidates, chasmfilling algorithm which eliminates black noises, determines the final cut pointsand separates originally attached traffic signs into individual ones. At each step,the mediate results as well as the efficiency in practice would be presented toshow the advantages and disadvantages of the developed algorithm. Thisreport concentrates on contour-based recognition of Swedish traffic signs. Thegeneral shapes cover upward triangle, downward triangle, circle, rectangle andoctagon. At last, a demonstration program would be presented to show howthe algorithm works in real-time environment.

Real-Time Recognition System for Traffic Signs

The aim of this thesis project is to develop the Traffic Sign Recognition algorithm for real time. Inreal time environment, vehicles move at high speed on roads. For the vehicle intelligent system itbecomes essential to detect, process and recognize the traffic sign which is coming in front ofvehicle with high relative velocity, at the right time, so that the driver would be able to pro-actsimultaneously on instructions given in the Traffic Sign. The system assists drivers about trafficsigns they did not recognize before passing them. With the Traffic Sign Recognition system, thevehicle becomes aware of the traffic environment and reacts according to the situation.The objective of the project is to develop a system which can recognize the traffic signs in real time.The three target parameters are the system’s response time in real-time video streaming, the trafficsign recognition speed in still images and the recognition accuracy. The system consists of threeprocesses; the traffic sign detection, the traffic sign recognition and the traffic sign tracking. Thedetection process uses physical properties of traffic signs based on a priori knowledge to detect roadsigns. It generates the road sign image as the input to the recognition process. The recognitionprocess is implemented using the Pattern Matching algorithm. The system was first tested onstationary images where it showed on average 97% accuracy with the average processing time of0.15 seconds for traffic sign recognition. This procedure was then applied to the real time videostreaming. Finally the tracking of traffic signs was developed using Blob tracking which showed theaverage recognition accuracy to 95% in real time and improved the system’s average response timeto 0.04 seconds. This project has been implemented in C-language using the Open Computer VisionLibrary.

QoS Evaluation of BandwidthSchedulers in IPTV Networks OfferedSRD Fluid Video Traffic

Internet protocol TV (IPTV) is predicted to be the key technology winner in the future. Efforts to accelerate the deployment of IPTV centralized model which is combined of VHO, encoders, controller, access network and Home network. Regardless of whether the network is delivering live TV, VOD, or Time-shift TV, all content and network traffic resulting from subscriber requests must traverse the entire network from the super-headend all the way to each subscriber's Set-Top Box (STB).IPTV services require very stringent QoS guarantees When IPTV traffic shares the network resources with other traffic like data and voice, how to ensure their QoS and efficiently utilize the network resources is a key and challenging issue. For QoS measured in the network-centric terms of delay jitter, packet losses and bounds on delay. The main focus of this thesis is on the optimized bandwidth allocation and smooth datatransmission. The proposed traffic model for smooth delivering video service IPTV network with its QoS performance evaluation. According to Maglaris et al [5] First, analyze the coding bit rate of a single video source. Various statistical quantities are derived from bit rate data collected with a conditional replenishment inter frame coding scheme. Two correlated Markov process models (one in discrete time and one incontinuous time) are shown to fit the experimental data and are used to model the input rates of several independent sources into a statistical multiplexer. Preventive control mechanism which is to be include CAC, traffic policing used for traffic control.QoS has been evaluated of common bandwidth scheduler( FIFO) by use fluid models with Markovian queuing method and analysis the result by using simulator andanalytically, Which is measured the performance of the packet loss, overflow and mean waiting time among the network users.

QoS evaluation of bandwidth schedulersin IPTV networks offered SRD fluidvideo traffic: a simulation study

IPTV is now offered by several operators in Europe, US and Asia using broadcast video over private IP networks that are isolated from Internet. IPTV services rely ontransmission of live (real-time) video and/or stored video. Video on Demand (VoD)and Time-shifted TV are implemented by IP unicast and Broadcast TV (BTV) and Near video on demand are implemented by IP multicast. IPTV services require QoS guarantees and can tolerate no more than 10-6 packet loss probability, 200 ms delay, and 50 ms jitter. Low delay is essential for satisfactory trick mode performance(pause, resume,fast forward) for VoD, and fast channel change time for BTV. Internet Traffic Engineering (TE) is defined in RFC 3272 and involves both capacity management and traffic management. Capacity management includes capacityplanning, routing control, and resource management. Traffic management includes (1)nodal traffic control functions such as traffic conditioning, queue management, scheduling, and (2) other functions that regulate traffic flow through the network orthat arbitrate access to network resources. An IPTV network architecture includes multiple networks (core network, metronetwork, access network and home network) that connects devices (super head-end, video hub office, video serving office, home gateway, set-top box). Each IP router in the core and metro networks implements some queueing and packet scheduling mechanism at the output link controller. Popular schedulers in IP networks include Priority Queueing (PQ), Class-Based Weighted Fair Queueing (CBWFQ), and Low Latency Queueing (LLQ) which combines PQ and CBWFQ.The thesis analyzes several Packet Scheduling algorithms that can optimize the tradeoff between system capacity and end user performance for the traffic classes. Before in the simulator FIFO,PQ,GPS queueing methods were implemented inside. This thesis aims to implement the LLQ scheduler inside the simulator and to evaluate the performance of these packet schedulers. The simulator is provided by ErnstNordström and Simulator was built in Visual C++ 2008 environmentand tested and analyzed in MatLab 7.0 under windows VISTA.

An Upgrade of Network Traffic Recognition System for SIP/VoIP Traffic Recognition

The purpose of this project is to update the tool of Network Traffic Recognition System (NTRS) which is proprietary software of Ericsson AB and Tsinghua University, and to implement the updated tool to finish SIP/VoIP traffic recognition. Basing on the original NTRS, I analyze the traffic recognition principal of NTRS, and redesign the structure and module of the tool according to characteristics of SIP/VoIP traffic, and then finally I program to achieve the upgrade. After the final test with our SIP data trace files in the updated system, a satisfactory result is derived. The result presents that our updated system holds a rate of recognition on a confident level in the SIP session recognition as well as the VoIP call recognition. In the comparison with the software of Wireshark, our updated system has a result which is extremely close to Wireshark’s output, and the working time is much less than Wireshark. In the aspect of practicability, the memory overflow problem is avoided, and the updated system can output the specific information of SIP/VoIP traffic recognition, such as SIP type, SIP state, VoIP state, etc. The upgrade fulfills the demand of this project.

Analysis of EnterpriseVoIP Traffic from aWire line IMS System

With the rapid development of telecommunication industry, the IP multimedia Subsystem (IMS) could very well be the panacea for most telecom operators. It is originally defined as the core network for 3G mobile systems by the 3rd Generation Partnership Project (3GPP), the more recent development is merging between fixed line network and wireless networkd This report researchs the characteristic of the IMS data and proposes an IMS characterization analysis. We captured the IMS traffic data with 10 tousands users for about 41 hours. By analyzing the characteristics of the IMS, we know that the most important application in the IMS is VoIP call. Then we use the tool designed by Tsinghua University & Ericsson Company to recognize the data, and the results we got can be used to build the traffic models. From the results of the traffic models, I will get some reasons and conclusion. The traffic model gives out the types of session and types of VoIP call. I bring into a concept—busy hour. This concept is very important because it can help us to know which period is the peak of the VoIP call. The busy hour is from 10:00 to 11:00 in the morning. I also bring into another concept—connection ratio. This concept is significant because it can evaluate whether the VoIP call is good when it use IMS network. By comparing the traffic model with other one’s models, we found the different results from them, both the accuracy and the busy hour. From the contract, we got the advantages of our traffic models.