Mechanism of mitosis-specific activation of MEK1

Activation of cyclin B-Cdc2 is an absolute requirement for entry into mitosis, but other protein kinase pathways that also have mitotic functions are activated during G(2)/M progression. The MAPK cascade has well established roles in entry and exit from mitosis in Xenopus, but relatively little is known about the regulation and function of this pathway in mammalian mitosis. Here we report a detailed analysis of the activity of all components of the Ras/Raf/MEK/ERK pathway in HeLa cells during normal G(2)/M. The focus of this pathway is the dramatic activation of an endomembrane-associated MEK1 without the corresponding activation of the MEK substrate ERK. This is because of the uncoupling of MEK1 activation from ERK activation. The mechanism of this uncoupling involves the cyclin B-Cdc2-dependent proteolytic cleavage of the N-terminal ERK-binding domain of MEK1 and the phosphorylation of Thr(286). These results demonstrate that cyclin B-Cdc2 activity regulates signaling through the MAPK pathway in mitosis.

A molecular mechanism for mRNA trafficking in neuronal dendrites

Specific neuronal mRNAs are localized in dendrites, often concentrated in dendritic spines and spine synapses, where they are translated. The molecular mechanism of localization is mostly unknown. Here we have explored the roles of A2 response element (A2RE), a cis-acting signal for oligodendrocyte RNA trafficking, and its cognate trans-acting factor, heterogeneous nuclear ribonucleoprotein ( hnRNP) A2, in neurons. Fluorescently labeled chimeric RNAs containing A2RE were microinjected into hippocampal neurons, and RNA transport followed using confocal laser scanning microscopy. These RNA molecules, but not RNA lacking the A2RE sequence, were transported in granules to the distal neurites. hnRNP A2 protein was implicated as the cognate trans-acting factor: it was colocalized with RNA in cytoplasmic granules, and RNA trafficking in neurites was compromised by A2RE mutations that abrogate hnRNP A2 binding. Coinjection of antibodies to hnRNP A2 halved the number of trafficking cells, and treatment of neurons with antisense oligonucleotides also disrupted A2RE - RNA transport. Colchicine inhibited trafficking, whereas cells treated with cytochalasin were unaffected, implicating involvement of microtubules rather than microfilaments. A2RE-like sequences are found in a subset of dendritically localized mRNAs, which, together with these results, suggests that a molecular mechanism based on this cis-acting sequence may contribute to dendritic RNA localization.

Negotiation mechanism for self-organized scheduling system

This paper presents a negotiation mechanism for Dynamic Scheduling based on Swarm Intelligence (SI). Under the new negotiation mechanism, agents must compete to obtain a global schedule. SI is the general term for several computational techniques which use ideas and get inspiration from the social behaviors of insects and other animals. This work is concerned with negotiation, the process through which multiple selfinterested agents can reach agreement over the exchange of operations on competitive resources.

Cooperation mechanism for team-work based multi-agent system in dynamic scheduling through meta-heuristics

This paper describes a Multi-agent Scheduling System that assumes the existence of several Machines Agents (which are decision-making entities) distributed inside the Manufacturing System that interact and cooperate with other agents in order to obtain optimal or near-optimal global performances. Agents have to manage their internal behaviors and their relationships with other agents via cooperative negotiation in accordance with business policies defined by the user manager. Some Multi Agent Systems (MAS) organizational aspects are considered. An original Cooperation Mechanism for a Team-work based Architecture is proposed to address dynamic scheduling using Meta-Heuristics.

Electrochemical and spectroscopic studies of the oxidation mechanism of the herbicide propanil

Electrochemical oxidation of propanil in deuterated solutions was studied by cyclic, differential pulse, and square wave voltammetry using a glassy carbon microelectrode. The oxidation of propanil in deuterated acid solutions occurs at the nitrogen atom of the amide at a potential of +1.15 V vs Ag/ AgCl. It was also found that, under the experimental conditions used, protonation at the oxygen atom of propanil occurs, leading to the appearance of another species in solution which oxidizes at +0.60 V. The anodic peak found at +0.79 V vs Ag/AgCl in deuterated basic solutions is related to the presence of an anionic species in which a negative charge is on the nitrogen atom. The electrochemical data were confirmed by the identification of all the species formed in acidic and basic deuterated solutions by means of NMR spectroscopy. The results are supported by electrochemical and spectroscopic studies of acetanilide in deuterated solutions.

On the adaptation of broadcast transactions in token-passing fieldbus networks with heterogeneous transmission media

Broadcast networks that are characterised by having different physical layers (PhL) demand some kind of traffic adaptation between segments, in order to avoid traffic congestion in linking devices. In many LANs, this problem is solved by the actual linking devices, which use some kind of flow control mechanism that either tell transmitting stations to pause (the transmission) or just discard frames. In this paper, we address the case of token-passing fieldbus networks operating in a broadcast fashion and involving message transactions over heterogeneous (wired or wireless) physical layers. For the addressed case, real-time and reliability requirements demand a different solution to the traffic adaptation problem. Our approach relies on the insertion of an appropriate idle time before a station issuing a request frame. In this way, we guarantee that the linking devices’ queues do not increase in a way that the timeliness properties of the overall system turn out to be unsuitable for the targeted applications.

Transport mechanism in ionic liquid membranes and the study of thiomersal biodegradation

The initial goal of this work was the development of a supported liquid membrane (SLM) bioreactor for the remediation of vaccine production effluents contaminated with a highly toxic organomercurial – thiomersal. Therefore, two main aspects were focused on: 1) the development of a stable supported liquid membrane – using room temperature ionic liquids (RTILs) – for the selective transport of thiomersal from the wastewater to a biological compartment, 2) study of the biodegradation kinetics of thiomersal to metallic mercury by a Pseudomonas putida strain. The first part of the work focused on the evaluation of the physicochemical properties of ionic liquids and on the SLMs’ operational stability. The results obtained showed that, although it is possible to obtain a SLM with a high stability, water possesses nonnegligible solubility in the RTILs studied. The formation of water clusters inside the hydrophobic ionic liquid was identified and found to regulate the transport of water and small ions. In practical terms, this meant that, although it was possible to transport thiomersal from the vaccine effluent to the biological compartment, complete isolation of the microbial culture could not be guaranteed and the membrane might ultimately be permeable to other species present in the aqueous vaccine wastewater. It was therefore decided not to operate the initially targeted integrated system but, instead, the biological system by itself. Additionally, attention was given to the development of a thorough understanding of the transport mechanisms involved in the solubilisation and transport of water through supported liquid membranes with RTILs as well as to the evaluation of the effect of water uptake by the SLM in the transport mechanisms of water-soluble solutes and its effect on SLM performance. The results obtained highlighted the determinant role played by water – solubilised inside the ionic liquids – on the transport mechanism. It became clear that the transport mechanism of water and water-soluble solutes through SLMs with [CnMIM][PF6] RTILs was regulated by the dynamics of water clusters inside the RTIL, rather than by molecular diffusion through the bulk of the ionic liquid. Although the stability tests vi performed showed that there were no significant losses of organic phase from the membrane pores, the formation of water clusters inside the ionic liquid, which constitute new, non-selective environments for solute transport, leads to a clear deterioration of SLM performance and selectivity. Nevertheless, electrical impedance spectroscopy characterisation of the SLMs showed that the formation of water clusters did not seem to have a detrimental effect on the SLMs’ electrical characteristics and highlighted the potential of using this type of membranes in electrochemical applications with low resistance requirements. The second part of the work studied the kinetics of thiomersal degradation by a pure culture of P. putida spi3 strain, in batch culture and using a synthe tic wastewater. A continuous ly stirred tank reactor fed with the synthetic wastewater was also operated and the bioreactor’s performance and robustness, when exposed to thiomersal shock loads, were evaluated. Finally, a bioreactor for the biological treatment of a real va ccine production effluent was set up and operated at different dilution rates. Thus it was possible to treat a real thiomersal-contaminated effluent, lowering the outlet mercury concentration to values below the European limit for mercury effluent discharges.

Smart-HOP: a reliable handoff mechanism for mobile wireless sensor networks

Handoff processes, the events where mobile nodes select the best access point available to transfer data, have been well studied in cellular and WiFi networks. However, wireless sensor networks (WSN) pose a new set of challenges due to their simple low-power radio transceivers and constrained resources. This paper proposes smart-HOP, a handoff mechanism tailored for mobile WSN applications. This work provides two important contributions. First, it demonstrates the intrinsic relationship between handoffs and the transitional region. The evaluation shows that handoffs perform the best when operating in the transitional region, as opposed to operating in the more reliable connected region. Second, the results reveal that a proper fine tuning of the parameters, in the transitional region, can reduce handoff delays by two orders of magnitude, from seconds to tens of milliseconds.

Entity localization and tracking: a sensor fusion-based mechanism in WSNs

Knowing exactly where a mobile entity is and monitoring its trajectory in real-time has recently attracted a lot of interests from both academia and industrial communities, due to the large number of applications it enables, nevertheless, it is nowadays one of the most challenging problems from scientific and technological standpoints. In this work we propose a tracking system based on the fusion of position estimations provided by different sources, that are combined together to get a final estimation that aims at providing improved accuracy with respect to those generated by each system individually. In particular, exploiting the availability of a Wireless Sensor Network as an infrastructure, a mobile entity equipped with an inertial system first gets the position estimation using both a Kalman Filter and a fully distributed positioning algorithm (the Enhanced Steepest Descent, we recently proposed), then combines the results using the Simple Convex Combination algorithm. Simulation results clearly show good performance in terms of the final accuracy achieved. Finally, the proposed technique is validated against real data taken from an inertial sensor provided by THALES ITALIA.

TDBS: a time division beacon scheduling mechanism for ZigBee cluster-tree wireless sensor networks

Synchronization is a challenging and important issue for time-sensitive Wireless Sensor Networks (WSN) since it requires a mutual spatiotemporal coordination between the nodes. In that concern, the IEEE 802.15.4/ZigBee protocols embody promising technologies for WSNs, but are still ambiguous on how to efficiently build synchronized multiple-cluster networks, specifically for the case of cluster-tree topologies. In fact, the current IEEE 802.15.4/ZigBee specifications restrict the synchronization to beacon-enabled (by the generation of periodic beacon frames) star networks, while they support multi-hop networking in mesh topologies, but with no synchronization. Even though both specifications mention the possible use of cluster-tree topologies, which combine multi-hop and synchronization features, the description on how to effectively construct such a network topology is missing. This paper tackles this issue by unveiling the ambiguities regarding the use of the cluster-tree topology and proposing a synchronization mechanism based on Time Division Beacon Scheduling (TDBS) to build cluster-tree WSNs. In addition, we propose a methodology for efficiently managing duty-cycles in every cluster, ensuring the fairest use of bandwidth resources. The feasibility of the TDBS mechanism is clearly demonstrated through an experimental test-bed based on our open-source implementation of the IEEE 802.15.4/ZigBee protocols.

An implicit GTS allocation mechanism in IEEE 802.15.4 for time-sensitive wireless sensor networks: theory and practice

Timeliness guarantee is an important feature of the recently standardized IEEE 802.15.4 protocol, turning it quite appealing for Wireless Sensor Network (WSN) applications under timing constraints. When operating in beacon-enabled mode, this protocol allows nodes with real-time requirements to allocate Guaranteed Time Slots (GTS) in the contention-free period. The protocol natively supports explicit GTS allocation, i.e. a node allocates a number of time slots in each superframe for exclusive use. The limitation of this explicit GTS allocation is that GTS resources may quickly disappear, since a maximum of seven GTSs can be allocated in each superframe, preventing other nodes to benefit from guaranteed service. Moreover, the GTS may be underutilized, resulting in wasted bandwidth. To overcome these limitations, this paper proposes i-GAME, an implicit GTS Allocation Mechanism in beacon-enabled IEEE 802.15.4 networks. The allocation is based on implicit GTS allocation requests, taking into account the traffic specifications and the delay requirements of the flows. The i-GAME approach enables the use of one GTS by multiple nodes, still guaranteeing that all their (delay, bandwidth) requirements are satisfied. For that purpose, we propose an admission control algorithm that enables to decide whether to accept a new GTS allocation request or not, based not only on the remaining time slots, but also on the traffic specifications of the flows, their delay requirements and the available bandwidth resources. We show that our approach improves the bandwidth utilization as compared to the native explicit allocation mechanism defined in the IEEE 802.15.4 standard. We also present some practical considerations for the implementation of i-GAME, ensuring backward compatibility with the IEEE 801.5.4 standard with only minor add-ons. Finally, an experimental evaluation on a real system that validates our theoretical analysis and demonstrates the implementation of i-GAME is also presented

H-NAMe: specifying, implementing and testing a hidden-node avoidance mechanism for wireless sensor networks

The hidden-node problem has been shown to be a major source of Quality-of-Service (QoS) degradation in Wireless Sensor Networks (WSNs) due to factors such as the limited communication range of sensor nodes, link asymmetry and the characteristics of the physical environment. In wireless contention-based Medium Access Control protocols, if two nodes that are not visible to each other transmit to a third node that is visible to the formers, there will be a collision – usually called hidden-node or blind collision. This problem greatly affects network throughput, energy-efficiency and message transfer delays, which might be particularly dramatic in large-scale WSNs. This technical report tackles the hidden-node problem in WSNs and proposes HNAMe, a simple yet efficient distributed mechanism to overcome it. H-NAMe relies on a grouping strategy that splits each cluster of a WSN into disjoint groups of non-hidden nodes and then scales to multiple clusters via a cluster grouping strategy that guarantees no transmission interference between overlapping clusters. We also show that the H-NAMe mechanism can be easily applied to the IEEE 802.15.4/ZigBee protocols with only minor add-ons and ensuring backward compatibility with the standard specifications. We demonstrate the feasibility of H-NAMe via an experimental test-bed, showing that it increases network throughput and transmission success probability up to twice the values obtained without H-NAMe. We believe that the results in this technical report will be quite useful in efficiently enabling IEEE 802.15.4/ZigBee as a WSN protocol.

A time division beacon scheduling mechanism for IEEE 802.15.4/Zigbee cluster-tree wireless sensor networks

While the IEEE 802.15.4/Zigbee protocol stack is being considered as a promising technology for low-cost low-power Wireless Sensor Networks (WSNs), several issues in the standard specifications are still open. One of those ambiguous issues is how to build a synchronized multi-hop cluster-tree network, which is quite suitable for ensuring QoS support in WSNs. In fact, the current IEEE 802.15.4/Zigbee specifications restrict the synchronization in the beacon-enabled mode (by the generation of periodic beacon frames) to star-based networks, while it supports multi-hop networking using the peer-to-peer mesh topology, but with no synchronization. Even though both specifications mention the possible use of cluster-tree topologies, which combine multihop and synchronization features, the description on how to effectively construct such a network topology is missing. This paper tackles this problem, unveils the ambiguities regarding the use of the cluster-tree topology and proposes a synchronization mechanism based on Time Division Beacon Scheduling to construct cluster-tree WSNs. We also propose a methodology for an efficient duty cycle management in each router (cluster-head) of a cluster-tree WSN that ensures the fairest use of bandwidth resources. The feasibility of the proposal is clearly demonstrated through an experimental test bed based on our own implementation of the IEEE 802.15.4/Zigbee protocol.

A simulation model for the IEEE 802.15.4 protocol: delay/throughput evaluation of the GTS mechanism

The IEEE 802.15.4 protocol has the ability to support time-sensitive Wireless Sensor Network (WSN) applications due to the Guaranteed Time Slot (GTS) Medium Access Control mechanism. Recently, several analytical and simulation models of the IEEE 802.15.4 protocol have been proposed. Nevertheless, currently available simulation models for this protocol are both inaccurate and incomplete, and in particular they do not support the GTS mechanism. In this paper, we propose an accurate OPNET simulation model, with focus on the implementation of the GTS mechanism. The motivation that has driven this work is the validation of the Network Calculus based analytical model of the GTS mechanism that has been previously proposed and to compare the performance evaluation of the protocol as given by the two alternative approaches. Therefore, in this paper we contribute an accurate OPNET model for the IEEE 802.15.4 protocol. Additionally, and probably more importantly, based on the simulation model we propose a novel methodology to tune the protocol parameters such that a better performance of the protocol can be guaranteed, both concerning maximizing the throughput of the allocated GTS as well as concerning minimizing frame delay.

Engineering a search and rescue application with a wireless sensor network - based localization mechanism

The advent of Wireless Sensor Network (WSN) technologies is paving the way for a panoply of new ubiquitous computing applications, some of them with critical requirements. In the ART-WiSe framework, we are designing a two-tiered communication architecture for supporting real-time and reliable communications in WSNs. Within this context, we have been developing a test-bed application, for testing, validating and demonstrating our theoretical findings - a search&rescue/pursuit-evasion application. Basically, a WSN deployment is used to detect, localize and track a target robot and a station controls a rescuer/pursuer robot until it gets close enough to the target robot. This paper describes how this application was engineered, particularly focusing on the implementation of the localization mechanism.