Recombinant protein production in yeast:methods and protocols

In the last few years, significant advances have been made in understanding how a yeast cell responds to the stress of producing a recombinant protein, and how this information can be used to engineer improved host strains. The molecular biology of the expression vector, through the choice of promoter, tag and codon optimization of the target gene, is also a key determinant of a high-yielding protein production experiment. Recombinant Protein Production in Yeast: Methods and Protocols examines the process of preparation of expression vectors, transformation to generate high-yielding clones, optimization of experimental conditions to maximize yields, scale-up to bioreactor formats and disruption of yeast cells to enable the isolation of the recombinant protein prior to purification. Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.

A new three phase method (SDP method) for the multi-objective vehicle routing problem with simultaneous delvery and pickup (VRPSDP)

Transportation service operators are witnessing a growing demand for bi-directional movement of goods. Given this, the following thesis considers an extension to the vehicle routing problem (VRP) known as the delivery and pickup transportation problem (DPP), where delivery and pickup demands may occupy the same route. The problem is formulated here as the vehicle routing problem with simultaneous delivery and pickup (VRPSDP), which requires the concurrent service of the demands at the customer location. This formulation provides the greatest opportunity for cost savings for both the service provider and recipient. The aims of this research are to propose a new theoretical design to solve the multi-objective VRPSDP, provide software support for the suggested design and validate the method through a set of experiments. A new real-life based multi-objective VRPSDP is studied here, which requires the minimisation of the often conflicting objectives: operated vehicle fleet size, total routing distance and the maximum variation between route distances (workload variation). The former two objectives are commonly encountered in the domain and the latter is introduced here because it is essential for real-life routing problems. The VRPSDP is defined as a hard combinatorial optimisation problem, therefore an approximation method, Simultaneous Delivery and Pickup method (SDPmethod) is proposed to solve it. The SDPmethod consists of three phases. The first phase constructs a set of diverse partial solutions, where one is expected to form part of the near-optimal solution. The second phase determines assignment possibilities for each sub-problem. The third phase solves the sub-problems using a parallel genetic algorithm. The suggested genetic algorithm is improved by the introduction of a set of tools: genetic operator switching mechanism via diversity thresholds, accuracy analysis tool and a new fitness evaluation mechanism. This three phase method is proposed to address the shortcoming that exists in the domain, where an initial solution is built only then to be completely dismantled and redesigned in the optimisation phase. In addition, a new routing heuristic, RouteAlg, is proposed to solve the VRPSDP sub-problem, the travelling salesman problem with simultaneous delivery and pickup (TSPSDP). The experimental studies are conducted using the well known benchmark Salhi and Nagy (1999) test problems, where the SDPmethod and RouteAlg solutions are compared with the prominent works in the VRPSDP domain. The SDPmethod has demonstrated to be an effective method for solving the multi-objective VRPSDP and the RouteAlg for the TSPSDP.

Developing Agent Interaction Protocols with PRALU

The purpose of the paper is to explore the possibility of applying existing formal theories of description and design of distributed and concurrent systems to interaction protocols for real-time multi-agent systems. In particular it is shown how the language PRALU, proposed for description of parallel logical control algorithms and rooted in the Petri net formalism, can be used for the modeling of complex concurrent conversations between agents in a multi-agent system. It is demonstrated with a known example of English auction on how to specify an agent interaction protocol using considered means.

Adaptive Routing and Multi-Agent Control for Information Flows in IP-Networks

The principles of adaptive routing and multi-agent control for information flows in IP-networks.

Neural Approach in Multi-Agent Routing for Static Telecommunication Networks

The problem of multi-agent routing in static telecommunication networks with fixed configuration is considered. The problem is formulated in two ways: for centralized routing schema with the coordinator-agent (global routing) and for distributed routing schema with independent agents (local routing). For both schemas appropriate Hopfield neural networks (HNN) are constructed.

Extending Object-Oriented Network Protocols via alternative Transportation Bindings (Web Services over XMPP)

Distributed and/or composite web applications are driven by intercommunication via web services, which employ application-level protocols, such as SOAP. However, these protocols usually rely on the classic HTTP for transportation. HTTP is quite efficient for what it does — delivering web page content, but has never been intended to carry complex web service oriented communication. Today there exist modern protocols that are much better fit for the job. Such a candidate is XMPP. It is an XML-based, asynchronous, open protocol that has built-in security and authentication mechanisms and utilizes a network of federated servers. Sophisticated asynchronous multi-party communication patterns can be established, effectively aiding web service developers. This paper’s purpose is to prove by facts, comparisons, and practical examples that XMPP is not only better suited than HTTP to serve as middleware for web service protocols, but can also contribute to the overall development state of web services.

Efficient algorithm for routing optimization via statistical mechanics

Many practical routing algorithms are heuristic, adhoc and centralized, rendering generic and optimal path configurations difficult to obtain. Here we study a scenario whereby selected nodes in a given network communicate with fixed routers and employ statistical physics methods to obtain optimal routing solutions subject to a generic cost. A distributive message-passing algorithm capable of optimizing the path configuration in real instances is devised, based on the analytical derivation, and is greatly simplified by expanding the cost function around the optimized flow. Good algorithmic convergence is observed in most of the parameter regimes. By applying the algorithm, we study and compare the pros and cons of balanced traffic configurations to that of consolidated traffic, which provides important implications to practical communication and transportation networks. Interesting macroscopic phenomena are observed from the optimized states as an interplay between the communication density and the cost functions used. © 2013 IEEE.

Chain routing:a new routing framework for the Internet based on complete orders

A new framework to perform routing at the autonomous system (AS) level is proposed here. This mechanism, called chain routing framework (CRF), uses complete orders as its main topological unit. Since complete orders are acyclic digraphs that possess a known topology, it is possible to use these acyclic structures to route consistently packets between a group of ASs. The adoption of complete orders also allows easy identification and avoidance of persistent route oscillations, eliminates the possibility of developing transient loops in paths and provides a structure that facilitates the implementation of traffic engineering. Moreover, by combining CRF with other mechanisms that implement complete orders in time, the authors propose that it is possible to design a new routing protocol, which can be more reliable and stable than the border gateway protocol. © 2011 The Institution of Engineering and Technology.

On the drug adsorption capacity of SBA-15 obtained from various detemplation protocols

The effect of the mild detemplation method, based on Fenton chemistry (with and without previous solvent extraction), and calcination was evaluated by the drug uptake capacity of SBA-15 materials. A number of characterization techniques were applied for evaluation and comparison of the materials properties such as TGA, CNH, N2 physisorption and 29Si NMR. The mild Fenton detemplation method rendered a nearly pristine SBA-15 without structural shrinkage, low residual template, improved surface area, pore volume and silanol concentration. The drug (ibuprofen) adsorption experiments were carried out by solution immersion in powdery form. The mild detemplated samples experienced an enhanced uptake that could be explained by the enhanced density of silanols (mmol/g), originated from the absence of calcination in the Fenton approaches. © 2014 Elsevier B.V.

Secure routing and trust computation in multihop infrastructureless networks

Today's wireless networks rely mostly on infrastructural support for their operation. With the concept of ubiquitous computing growing more popular, research on infrastructureless networks have been rapidly growing. However, such types of networks face serious security challenges when deployed. This dissertation focuses on designing a secure routing solution and trust modeling for these infrastructureless networks. ^ The dissertation presents a trusted routing protocol that is capable of finding a secure end-to-end route in the presence of malicious nodes acting either independently or in collusion, The solution protects the network from active internal attacks, known to be the most severe types of attacks in an ad hoc application. Route discovery is based on trust levels of the nodes, which need to be dynamically computed to reflect the malicious behavior in the network. As such, we have developed a trust computational model in conjunction with the secure routing protocol that analyzes the different malicious behavior and quantifies them in the model itself. Our work is the first step towards protecting an ad hoc network from colluding internal attack. To demonstrate the feasibility of the approach, extensive simulation has been carried out to evaluate the protocol efficiency and scalability with both network size and mobility. ^ This research has laid the foundation for developing a variety of techniques that will permit people to justifiably trust the use of ad hoc networks to perform critical functions, as well as to process sensitive information without depending on any infrastructural support and hence will enhance the use of ad hoc applications in both military and civilian domains. ^

Autonomic and embedded wireless sensor protocols for critical infrastructures

The tragic events of September 11th ushered a new era of unprecedented challenges. Our nation has to be protected from the alarming threats of adversaries. These threats exploit the nation's critical infrastructures affecting all sectors of the economy. There is the need for pervasive monitoring and decentralized control of the nation's critical infrastructures. The communications needs of monitoring and control of critical infrastructures was traditionally catered for by wired communication systems. These technologies ensured high reliability and bandwidth but are however very expensive, inflexible and do not support mobility and pervasive monitoring. The communication protocols are Ethernet-based that used contention access protocols which results in high unsuccessful transmission and delay. An emerging class of wireless networks, named embedded wireless sensor and actuator networks has potential benefits for real-time monitoring and control of critical infrastructures. The use of embedded wireless networks for monitoring and control of critical infrastructures requires secure, reliable and timely exchange of information among controllers, distributed sensors and actuators. The exchange of information is over shared wireless media. However, wireless media is highly unpredictable due to path loss, shadow fading and ambient noise. Monitoring and control applications have stringent requirements on reliability, delay and security. The primary issue addressed in this dissertation is the impact of wireless media in harsh industrial environment on the reliable and timely delivery of critical data. In the first part of the dissertation, a combined networking and information theoretic approach was adopted to determine the transmit power required to maintain a minimum wireless channel capacity for reliable data transmission. The second part described a channel-aware scheduling scheme that ensured efficient utilization of the wireless link and guaranteed delay. Various analytical evaluations and simulations are used to evaluate and validate the feasibility of the methodologies and demonstrate that the protocols achieved reliable and real-time data delivery in wireless industrial networks.

Trust based security mechanisms for wireless sensor networks

Wireless sensor networks are emerging as effective tools in the gathering and dissemination of data. They can be applied in many fields including health, environmental monitoring, home automation and the military. Like all other computing systems it is necessary to include security features, so that security sensitive data traversing the network is protected. However, traditional security techniques cannot be applied to wireless sensor networks. This is due to the constraints of battery power, memory, and the computational capacities of the miniature wireless sensor nodes. Therefore, to address this need, it becomes necessary to develop new lightweight security protocols. This dissertation focuses on designing a suite of lightweight trust-based security mechanisms and a cooperation enforcement protocol for wireless sensor networks. This dissertation presents a trust-based cluster head election mechanism used to elect new cluster heads. This solution prevents a major security breach against the routing protocol, namely, the election of malicious or compromised cluster heads. This dissertation also describes a location-aware, trust-based, compromise node detection, and isolation mechanism. Both of these mechanisms rely on the ability of a node to monitor its neighbors. Using neighbor monitoring techniques, the nodes are able to determine their neighbors’ reputation and trust level through probabilistic modeling. The mechanisms were designed to mitigate internal attacks within wireless sensor networks. The feasibility of the approach is demonstrated through extensive simulations. The dissertation also addresses non-cooperation problems in multi-user wireless sensor networks. A scalable lightweight enforcement algorithm using evolutionary game theory is also designed. The effectiveness of this cooperation enforcement algorithm is validated through mathematical analysis and simulation. This research has advanced the knowledge of wireless sensor network security and cooperation by developing new techniques based on mathematical models. By doing this, we have enabled others to build on our work towards the creation of highly trusted wireless sensor networks. This would facilitate its full utilization in many fields ranging from civilian to military applications.

Modeling security and cooperation in wireless networks using game theory

This research involves the design, development, and theoretical demonstration of models resulting in integrated misbehavior resolution protocols for ad hoc networked devices. Game theory was used to analyze strategic interaction among independent devices with conflicting interests. Packet forwarding at the routing layer of autonomous ad hoc networks was investigated. Unlike existing reputation based or payment schemes, this model is based on repeated interactions. To enforce cooperation, a community enforcement mechanism was used, whereby selfish nodes that drop packets were punished not only by the victim, but also by all nodes in the network. Then, a stochastic packet forwarding game strategy was introduced. Our solution relaxed the uniform traffic demand that was pervasive in other works. To address the concerns of imperfect private monitoring in resource aware ad hoc networks, a belief-free equilibrium scheme was developed that reduces the impact of noise in cooperation. This scheme also eliminated the need to infer the private history of other nodes. Moreover, it simplified the computation of an optimal strategy. The belief-free approach reduced the node overhead and was easily tractable. Hence it made the system operation feasible. Motivated by the versatile nature of evolutionary game theory, the assumption of a rational node is relaxed, leading to the development of a framework for mitigating routing selfishness and misbehavior in Multi hop networks. This is accomplished by setting nodes to play a fixed strategy rather than independently choosing a rational strategy. A range of simulations was carried out that showed improved cooperation between selfish nodes when compared to older results. Cooperation among ad hoc nodes can also protect a network from malicious attacks. In the absence of a central trusted entity, many security mechanisms and privacy protections require cooperation among ad hoc nodes to protect a network from malicious attacks. Therefore, using game theory and evolutionary game theory, a mathematical framework has been developed that explores trust mechanisms to achieve security in the network. This framework is one of the first steps towards the synthesis of an integrated solution that demonstrates that security solely depends on the initial trust level that nodes have for each other.^

Novel online data cleaning protocols for data streams in trajectory, Wireless Sensor Networks

The promise of Wireless Sensor Networks (WSNs) is the autonomous collaboration of a collection of sensors to accomplish some specific goals which a single sensor cannot offer. Basically, sensor networking serves a range of applications by providing the raw data as fundamentals for further analyses and actions. The imprecision of the collected data could tremendously mislead the decision-making process of sensor-based applications, resulting in an ineffectiveness or failure of the application objectives. Due to inherent WSN characteristics normally spoiling the raw sensor readings, many research efforts attempt to improve the accuracy of the corrupted or "dirty" sensor data. The dirty data need to be cleaned or corrected. However, the developed data cleaning solutions restrict themselves to the scope of static WSNs where deployed sensors would rarely move during the operation. Nowadays, many emerging applications relying on WSNs need the sensor mobility to enhance the application efficiency and usage flexibility. The location of deployed sensors needs to be dynamic. Also, each sensor would independently function and contribute its resources. Sensors equipped with vehicles for monitoring the traffic condition could be depicted as one of the prospective examples. The sensor mobility causes a transient in network topology and correlation among sensor streams. Based on static relationships among sensors, the existing methods for cleaning sensor data in static WSNs are invalid in such mobile scenarios. Therefore, a solution of data cleaning that considers the sensor movements is actively needed. This dissertation aims to improve the quality of sensor data by considering the consequences of various trajectory relationships of autonomous mobile sensors in the system. First of all, we address the dynamic network topology due to sensor mobility. The concept of virtual sensor is presented and used for spatio-temporal selection of neighboring sensors to help in cleaning sensor data streams. This method is one of the first methods to clean data in mobile sensor environments. We also study the mobility pattern of moving sensors relative to boundaries of sub-areas of interest. We developed a belief-based analysis to determine the reliable sets of neighboring sensors to improve the cleaning performance, especially when node density is relatively low. Finally, we design a novel sketch-based technique to clean data from internal sensors where spatio-temporal relationships among sensors cannot lead to the data correlations among sensor streams.

Trust Based Security Mechanisms for Wireless Sensor Networks

Wireless sensor networks are emerging as effective tools in the gathering and dissemination of data. They can be applied in many fields including health, environmental monitoring, home automation and the military. Like all other computing systems it is necessary to include security features, so that security sensitive data traversing the network is protected. However, traditional security techniques cannot be applied to wireless sensor networks. This is due to the constraints of battery power, memory, and the computational capacities of the miniature wireless sensor nodes. Therefore, to address this need, it becomes necessary to develop new lightweight security protocols. This dissertation focuses on designing a suite of lightweight trust-based security mechanisms and a cooperation enforcement protocol for wireless sensor networks. This dissertation presents a trust-based cluster head election mechanism used to elect new cluster heads. This solution prevents a major security breach against the routing protocol, namely, the election of malicious or compromised cluster heads. This dissertation also describes a location-aware, trust-based, compromise node detection, and isolation mechanism. Both of these mechanisms rely on the ability of a node to monitor its neighbors. Using neighbor monitoring techniques, the nodes are able to determine their neighbors’ reputation and trust level through probabilistic modeling. The mechanisms were designed to mitigate internal attacks within wireless sensor networks. The feasibility of the approach is demonstrated through extensive simulations. The dissertation also addresses non-cooperation problems in multi-user wireless sensor networks. A scalable lightweight enforcement algorithm using evolutionary game theory is also designed. The effectiveness of this cooperation enforcement algorithm is validated through mathematical analysis and simulation. This research has advanced the knowledge of wireless sensor network security and cooperation by developing new techniques based on mathematical models. By doing this, we have enabled others to build on our work towards the creation of highly trusted wireless sensor networks. This would facilitate its full utilization in many fields ranging from civilian to military applications.