N-terminal acetylome analyses and functional insights of the N-terminal acetyltransferase NatB

Protein N-terminal acetylation (Nt-acetylation) is an important mediator of protein function, stability, sorting, and localization. Although the responsible enzymes are thought to be fairly well characterized, the lack of identified in vivo substrates, the occurrence of Nt-acetylation substrates displaying yet uncharacterized N-terminal acetyltransferase (NAT) specificities, and emerging evidence of posttranslational Nt-acetylation, necessitate the use of genetic models and quantitative proteomics. NatB, which targets Met-Glu-, Met-Asp-, and Met-Asn-starting protein N termini, is presumed to Nt-acetylate 15% of all yeast and 18% of all human proteins. We here report on the evolutionary traits of NatB from yeast to human and demonstrate that ectopically expressed hNatB in a yNatB-Δ yeast strain partially complements the natB-Δ phenotypes and partially restores the yNatB Nt-acetylome. Overall, combining quantitative N-terminomics with yeast studies and knockdown of hNatB in human cell lines, led to the unambiguous identification of 180 human and 110 yeast NatB substrates. Interestingly, these substrates included Met-Gln- N-termini, which are thus now classified as in vivo NatB substrates. We also demonstrate the requirement of hNatB activity for maintaining the structure and function of actomyosin fibers and for proper cellular migration. In addition, expression of tropomyosin-1 restored the altered focal adhesions and cellular migration defects observed in hNatB-depleted HeLa cells, indicative for the conserved link between NatB, tropomyosin, and actin cable function from yeast to human.

Applying a validation framework to a working airport terminal model

Validation is an important issue in the development and application of Bayesian Belief Network (BBN) models, especially when the outcome of the model cannot be directly observed. Despite this, few frameworks for validating BBNs have been proposed and fewer have been applied to substantive real-world problems. In this paper we adopt the approach by Pitchforth and Mengersen (2013), which includes nine validation tests that each focus on the structure, discretisation, parameterisation and behaviour of the BBNs included in the case study. We describe the process and result of implementing a validation framework on a model of a real airport terminal system with particular reference to its effectiveness in producing a valid model that can be used and understood by operational decision makers. In applying the proposed validation framework we demonstrate the overall validity of the Inbound Passenger Facilitation Model as well as the effectiveness of the validity framework itself.

Personal media and wireless cities : towards an urban spatial analysis

For the past decade, at least, varieties of small, hand held networked instruments have appeared on the global scene, selling in record numbers, and being utilized by all manner of persons from the old to the young; children, women, men, the wealthy and the poor and in all countries. Their presences bespeak a radical shift in telecommunications infrastructure and the future of communications. They are particularly visible in urban areas where mobile transmission network infrastructure (3G, 4G, cellular and Wi-Fi) is more established and substantial, options more plentiful, and density of populations more dramatic. These end user products—I phones, cell phones, Blackberries, DSi, DS, IPads, Zooms, and others – of the mobile communications industry are the latest, hottest globalized commodities. At the same time, wirelessness, or the state of being wireless, and therefore capable of taking along one's networks, communicating from unlikely spaces, and navigating with GPS, is a complex social, political and economic communications phenomenon of early 21st century life. This thesis examines the specter of being wireless in cities. It lends the entire idea an experimentally envisioned, historical and planned context wherein personalization of media tools is seen both as a design development of corporate, artistic, and military imagination, as well as a profound social phenomenon enabling new forms of sharing, belonging, and urban community. In doing that it asserts the parameters of a new mobile space which, aside from clear benefits to humankind by way of mobility, has reinscribed numerous categories including gender. Moreover, it posits the recognition of other, more nuanced theoretical spaces for complex readings of gender and gendered use, including some instantiation of the notion of 'network' itself as a cyborgian and gendered social form. Additionally, cities are studied as places where technology is not only quickly popularized, but is connected to larger political interests, such as the reading of data, tracking of information, and the new security culture. In so doing the work has been undertaken as an urban spatial analysis and experimental ethnography, utilizing architectural, feminist, techno-utopian, industrial and theoretical literatures as discursive underpinnings from whence understandings and interpretations of mobile space, the mobile office, networked mobility, and personal media have come, linking the space of cities to specific, pioneering urban public art projects in which voice, texting and MMS have been utilized in expressions of ubiquitous networks and urban history. Through numerous examples of techno art, the thesis discusses the 'wireless city' as an emerging cultural, socially constructed economic and spatial entity, both conceived and formed through historic processes of urbanization.

Effective channel state information (CSI) feedback for MIMO systems in wireless broadband communications

This research has analysed both reciprocity and feedback mechanisms in multi-antenna wireless systems. It has presented the basis of an effective CSI feedback mechanism that efficiently provides the transmitter with the minimum information to allow the accurate knowledge of a rapidly changing channel. The simulations have been conducted using MATLAB to measure the improvement when the channel is estimated at the receiver in a 2 X 2 multi-antenna system and compared to the case of perfect channel knowledge at the receiver.

The role and future challenges of wireless communication networks for cooperative autonomous city vehicles

This paper elaborates on the use of future wireless communication networks for autonomous city vehicles. After addressing the state of technology, the paper explains the autonomous vehicle control system architecture and the Cybercars-2 communication framework; it presents experimental tests of communication-based real-time decision making; and discusses potential applications for communication in order to improve the localization and perception abilities of autonomous vehicles in urban environments.

Effects of wireless sensor network uncertainties on output-only modal analysis employing merged data of multiple tests

The use of Wireless Sensor Networks (WSNs) for vibration-based Structural Health Monitoring (SHM) has become a promising approach due to many advantages such as low cost, fast and flexible deployment. However, inherent technical issues such as data asynchronicity and data loss have prevented these distinct systems from being extensively used. Recently, several SHM-oriented WSNs have been proposed and believed to be able to overcome a large number of technical uncertainties. Nevertheless, there is limited research verifying the applicability of those WSNs with respect to demanding SHM applications like modal analysis and damage identification. Based on a brief review, this paper first reveals that Data Synchronization Error (DSE) is the most inherent factor amongst uncertainties of SHM-oriented WSNs. Effects of this factor are then investigated on outcomes and performance of the most robust Output-only Modal Analysis (OMA) techniques when merging data from multiple sensor setups. The two OMA families selected for this investigation are Frequency Domain Decomposition (FDD) and data-driven Stochastic Subspace Identification (SSI-data) due to the fact that they both have been widely applied in the past decade. Accelerations collected by a wired sensory system on a large-scale laboratory bridge model are initially used as benchmark data after being added with a certain level of noise to account for the higher presence of this factor in SHM-oriented WSNs. From this source, a large number of simulations have been made to generate multiple DSE-corrupted datasets to facilitate statistical analyses. The results of this study show the robustness of FDD and the precautions needed for SSI-data family when dealing with DSE at a relaxed level. Finally, the combination of preferred OMA techniques and the use of the channel projection for the time-domain OMA technique to cope with DSE are recommended.

Detection of sinkhole attack in wireless sensor networks

Generally wireless sensor networks rely of many-to-one communication approach for data gathering. This approach is extremely susceptible to sinkhole attack, where an intruder attracts surrounding nodes with unfaithful routing information, and subsequently presents selective forwarding or change the data that carry through it. A sinkhole attack causes an important threat to sensor networks and it should be considered that the sensor nodes are mostly spread out in open areas and of weak computation and battery power. In order to detect the intruder in a sinkhole attack this paper suggests an algorithm which firstly finds a group of suspected nodes by analyzing the consistency of data. Then, the intruder is recognized efficiently in the group by checking the network flow information. The proposed algorithm's performance has been evaluated by using numerical analysis and simulations. Therefore, accuracy and efficiency of algorithm would be verified.

Security in wireless sensor networks : issues and challenges

Wireless Sensor Networks (WSNs) are employed in numerous applications in different areas including military, ecology, and health; for example, to control of important information like the personnel position in a building, as a result, WSNs need security. However, several restrictions such as low capability of computation, small memory, limited resources of energy, and the unreliable channels employ communication in using WSNs can cause difficulty in use of security and protection in WSNs. It is very essential to save WSNs from malevolent attacks in unfriendly situations. Such networks require security plan due to various limitations of resources and the prominent characteristics of a wireless sensor network which is a considerable challenge. This article is an extensive review about problems of WSNs security, which examined recently by researchers and a better understanding of future directions for WSN security.

Effective routing with utilisation of cross-layer information for industrial wireless sensor networks

This thesis focuses on providing reliable data transmissions in large-scale industrial wireless sensor networks through improving network layer protocols. It addresses three major problems: scalability, dynamic industrial environments and coexistence of multiple types of data traffic in a network. Theoretical developments are conducted, followed by simulation studies for verification of theoretic results. The approach proposed in this thesis has been shown to be effective for large-scale network implementation and to provide improved data transmission reliability for both periodic and sporadic traffic.

Role of the N-terminal region in G protein-coupled receptor functions : negative modulation revealed by 5-HT2B receptor polymorphisms

The putative role of the N-terminal region of rhodopsin-like 7 transmembrane biogenic amine receptors in agonist-induced signaling has not yet been clarified despite recent advances in 7 transmembrane receptor structural biology. Given the existence of N-terminal nonsynonymous polymorphisms (R6G;E42G) within the HTR2B gene in a drug-abusing population, we assessed whether these polymorphisms affect 5-hydroxytryptamine 2B (5-HT2B) receptor in vitro pharmacologic and coupling properties in transfected COS-7 cells. Modification of the 5-HT2B receptor N terminus by the R6G;E42G polymorphisms increases such agonist signaling pathways as inositol phosphate accumulation as assessed by either classic or operational models. The N-terminal R6G;E42G mutations of the 5-HT2B receptor also increase cell proliferation and slow its desensitization kinetics compared with the wild-type receptor, further supporting a role for the N terminus in transduction efficacy. Furthermore, by coexpressing a tethered wild-type 5-HT2B receptor N terminus with a 5-HT2B receptor bearing a N-terminal deletion, we were able to restore original coupling. This reversion to normal activity of a truncated 5-HT2B receptor by coexpression of the membrane-tethered wild-type 5-HT2B receptor N terminus was not observed using a membrane-tethered 5-HT2B receptor R6G;E42G N terminus. These data suggest that the N terminus exerts a negative control over basal as well as agonist-stimulated receptor activity that is lost in the R6G;E42G mutant. Our findings reveal a new and unanticipated role of the 5-HT2B receptor N terminus as a negative modulator, affecting both constitutive and agonist-stimulated activity. Moreover, our data caution against excluding the N terminus and extracellular loops in structural studies of this 7 transmembrane receptor family

Re-evaluation of the effect of smoking on the methylation of N-terminal valine in haemoglobin

In view of the established extrapulmonary cancer sites targeted by smoking a multiplicity of compounds, and mechanisms might be involved. It has been debated that smoking caused increased incidence of N-methylvaline at the N-terminus of haemoglobin. Because this could indicate a relevance of methylating nitrosamines in tobacco smoke, data are presented from an industrial cohort of 35 smokers and 21 non-smokers repeatedly monitored between 1994 and 1999. In general, N-methylvaline adduct levels in haemoglobin of smokers were approximately 50% higher than those of non-smokers. The smoking-induced methylation of haemoglobin is likely to be caused by dimethylnitrosamine (N-nitroso-dimethylamine), a major nitrosamine in side-stream tobacco smoke. The biomonitoring data emphasise the potential value of N-methylvaline as a smoking-related biomarker and call for intensified research on tobacco smoke compounds that lead to macromolecular methylation process.

Circulating fragments of N-Terminal Pro-B-Type Natriuretic Peptides in plasma of heart failure patients

BACKGROUND: The use of nonstandardized N-terminal pro-B-type natriuretic peptide (NT-proBNP) assays can contribute to the misdiagnosis of heart failure (HF). Moreover, there is yet to be established a common consensus regarding the circulating forms of NT-proBNP being used in current assays. We aimed to characterize and quantify the various forms of NT-proBNP in the circulation of HF patients. METHODS: Plasma samples were collected from HF patients (n = 20) at rest and stored at -80 degrees C. NT-proBNP was enriched from HF patient plasma by use of immunoprecipitation followed by mass spectrometric analysis. Customized homogeneous sandwich AlphaLISA (R) immunoassays were developed and validated to quantify 6 fragments of NT-proBNP. RESULTS: Mass spectrometry identified the presence of several N- and C-terminally processed forms of circulating NT-proBNP, with physiological proteolysis between Pro2-Leu3, Leu3-Gly4, Pro6-Gly7, and Pro75-Arg76. Consistent with this result, AlphaLISA immunoassays demonstrated that antibodies targeting the extreme N or C termini measured a low apparent concentration of circulating NT-proBNP. The apparent circulating NT-proBNP concentration was increased with antibodies targeting nonglycosylated and nonterminal epitopes (P < 0.05). CONCLUSIONS: In plasma collected from HF patients, immunoreactive NT-proBNP was present as multiple N- and C-terminally truncated fragments of the full length NT-proBNP molecule. Immunodetection of NT-proBNP was significantly improved with the use of antibodies that did not target these terminal regions. These findings support the development of a next generation NT-proBNP assay targeting nonterminal epitopes as well as avoiding the central glycosylated region of this molecule. (c) 2013 American Association for Clinical Chemistry

Doppler and Pathloss Characterization for Vehicle-to-Vehicle communications at 5.8 GHz

Due to significant increase in vehicular accident and traffic congestions, vehicle to vehicle (V2V) communication based on the intelligent transport system (ITS) was introduced. However, to carry out efficient design and implementation of a reliable vehicular communication systems,a deep knowledge of the propagation channel characteristics in different environments is crucial, in particular the Doppler and pathloss parameters. Therefore, this paper presents an empirical V2V channel characterization and measurement performed under realistic urban, suburban and highway driving conditions in Brisbane, Australia. Based on Lin Cheng statistical Doppler Model (LCDM), values for the RMS Doppler spread and coherence time due to time selective nature of V2V channels were presented. Also, based on Log-distance power law model, values for the mean pathloss exponent and the standard deviation of shadowing were reported for urban, suburban and highway environments. The V2V channel parameters can be useful to system designers for the purpose of evaluating, simulating and developing new protocols and systems.