Analysis of the mechanism and regulation of lactose transport and metabolism in Clostridium acetobutylicum ATCC 824.

Although the acetone-butanol-ethanol (ABE) fermentation of Clostridium acetobutylicum is currently uneconomic, the ability of the bacterium to metabolise a wide range of carbohydrates offers the potential for revival based on the use of cheap, low grade substrates. We have investigated the uptake and metabolism of lactose, the major sugar in industrial whey waste, by C. acetobutylicum ATCC 824. Lactose is taken up via a phosphoenolpyruvate (PEP)-dependent phosphotransferase system (PTS) comprising both soluble and membrane-associated components, and the resulting phosphorylated derivative is hydrolysed by a phospho--galactosidase. These activities are induced during growth on lactose, but are absent in glucose-grown cells. Analysis of the C. acetobutylicum genome sequence identified a gene system, lacRFEG, encoding a transcriptional regulator of the DeoR family, IIA and IICB components of a lactose PTS, and phospho--galactosidase. During growth in medium containing both glucose and lactose, C. acetobutylicum exhibited a classical diauxic growth, and the lac operon was not expressed until glucose was exhausted from the medium. The presence upstream of lacR of a potential catabolite responsive element (cre) encompassing the transcriptional start site is indicative of the mechanism of carbon catabolite repression characteristic of low-GC Gram-positive bacteria. A pathway for the uptake and metabolism of lactose by this industrially important organism is proposed.

Timidity: A useful emotional mechanism for robot control?

M J Neal and J Timmis. Timidity: A useful mechanism for robot control? Informatica - special issue on perception and emotion based control, 4(27):197-204, 2003.

An Hysteresis-Habituation mechanism for Sensorimotor Scaffolding

R. Gunstone and M.H. Lee ?An Hysteresis-Habituation Mechanism for Sensorimotor Scaffolding?, Proc. AISB?03 Second International Symposium on Imitation in Animals and Artifacts, 189-190, 2003.

Iterative vs Simultaneous Fuzzy Rule Induction

M. Galea and Q. Shen. Iterative vs Simultaneous Fuzzy Rule Induction. Proceedings of the 14th International Conference on Fuzzy Systems, pages 767-772.

User-Level Sandboxing: a Safe and Efficient Mechanism for Extensibility

Extensible systems allow services to be configured and deployed for the specific needs of individual applications. This paper describes a safe and efficient method for user-level extensibility that requires only minimal changes to the kernel. A sandboxing technique is described that supports multiple logical protection domains within the same address space at user-level. This approach allows applications to register sandboxed code with the system, that may be executed in the context of any process. Our approach differs from other implementations that require special hardware support, such as segmentation or tagged translation look-aside buffers (TLBs), to either implement multiple protection domains in a single address space, or to support fast switching between address spaces. Likewise, we do not require the entire system to be written in a type-safe language, to provide fine-grained protection domains. Instead, our user-level sandboxing technique requires only paged-based virtual memory support, and the requirement that extension code is written either in a type-safe language, or by a trusted source. Using a fast method of upcalls, we show how our sandboxing technique for implementing logical protection domains provides significant performance improvements over traditional methods of invoking user-level services. Experimental results show our approach to be an efficient method for extensibility, with inter-protection domain communication costs close to those of hardware-based solutions leveraging segmentation.

On the Relationship Between File Sizes, Transport Protocols, and Self-Similar Network Traffic

Recent measurements of local-area and wide-area traffic have shown that network traffic exhibits variability at a wide range of scales self-similarity. In this paper, we examine a mechanism that gives rise to self-similar network traffic and present some of its performance implications. The mechanism we study is the transfer of files or messages whose size is drawn from a heavy-tailed distribution. We examine its effects through detailed transport-level simulations of multiple TCP streams in an internetwork. First, we show that in a "realistic" client/server network environment i.e., one with bounded resources and coupling among traffic sources competing for resources the degree to which file sizes are heavy-tailed can directly determine the degree of traffic self-similarity at the link level. We show that this causal relationship is not significantly affected by changes in network resources (bottleneck bandwidth and buffer capacity), network topology, the influence of cross-traffic, or the distribution of interarrival times. Second, we show that properties of the transport layer play an important role in preserving and modulating this relationship. In particular, the reliable transmission and flow control mechanisms of TCP (Reno, Tahoe, or Vegas) serve to maintain the long-range dependency structure induced by heavy-tailed file size distributions. In contrast, if a non-flow-controlled and unreliable (UDP-based) transport protocol is used, the resulting traffic shows little self-similar characteristics: although still bursty at short time scales, it has little long-range dependence. If flow-controlled, unreliable transport is employed, the degree of traffic self-similarity is positively correlated with the degree of throttling at the source. Third, in exploring the relationship between file sizes, transport protocols, and self-similarity, we are also able to show some of the performance implications of self-similarity. We present data on the relationship between traffic self-similarity and network performance as captured by performance measures including packet loss rate, retransmission rate, and queueing delay. Increased self-similarity, as expected, results in degradation of performance. Queueing delay, in particular, exhibits a drastic increase with increasing self-similarity. Throughput-related measures such as packet loss and retransmission rate, however, increase only gradually with increasing traffic self-similarity as long as reliable, flow-controlled transport protocol is used.

Improved Tracking of Multiple Humans with Trajectory Prediction and Occlusion Modeling

A combined 2D, 3D approach is presented that allows for robust tracking of moving bodies in a given environment as observed via a single, uncalibrated video camera. Tracking is robust even in the presence of occlusions. Low-level features are often insufficient for detection, segmentation, and tracking of non-rigid moving objects. Therefore, an improved mechanism is proposed that combines low-level (image processing) and mid-level (recursive trajectory estimation) information obtained during the tracking process. The resulting system can segment and maintain the tracking of moving objects before, during, and after occlusion. At each frame, the system also extracts a stabilized coordinate frame of the moving objects. This stabilized frame is used to resize and resample the moving blob so that it can be used as input to motion recognition modules. The approach enables robust tracking without constraining the system to know the shape of the objects being tracked beforehand; although, some assumptions are made about the characteristics of the shape of the objects, and how they evolve with time. Experiments in tracking moving people are described.

3D Trajectory Recovery for Tracking Multiple Objects and Trajectory Guided Recognition of Actions

A mechanism is proposed that integrates low-level (image processing), mid-level (recursive 3D trajectory estimation), and high-level (action recognition) processes. It is assumed that the system observes multiple moving objects via a single, uncalibrated video camera. A novel extended Kalman filter formulation is used in estimating the relative 3D motion trajectories up to a scale factor. The recursive estimation process provides a prediction and error measure that is exploited in higher-level stages of action recognition. Conversely, higher-level mechanisms provide feedback that allows the system to reliably segment and maintain the tracking of moving objects before, during, and after occlusion. The 3D trajectory, occlusion, and segmentation information are utilized in extracting stabilized views of the moving object. Trajectory-guided recognition (TGR) is proposed as a new and efficient method for adaptive classification of action. The TGR approach is demonstrated using "motion history images" that are then recognized via a mixture of Gaussian classifier. The system was tested in recognizing various dynamic human outdoor activities; e.g., running, walking, roller blading, and cycling. Experiments with synthetic data sets are used to evaluate stability of the trajectory estimator with respect to noise.

Trajectory Guided Tracking and Recognition of Actions

A combined 2D, 3D approach is presented that allows for robust tracking of moving people and recognition of actions. It is assumed that the system observes multiple moving objects via a single, uncalibrated video camera. Low-level features are often insufficient for detection, segmentation, and tracking of non-rigid moving objects. Therefore, an improved mechanism is proposed that integrates low-level (image processing), mid-level (recursive 3D trajectory estimation), and high-level (action recognition) processes. A novel extended Kalman filter formulation is used in estimating the relative 3D motion trajectories up to a scale factor. The recursive estimation process provides a prediction and error measure that is exploited in higher-level stages of action recognition. Conversely, higher-level mechanisms provide feedback that allows the system to reliably segment and maintain the tracking of moving objects before, during, and after occlusion. The 3D trajectory, occlusion, and segmentation information are utilized in extracting stabilized views of the moving object that are then used as input to action recognition modules. Trajectory-guided recognition (TGR) is proposed as a new and efficient method for adaptive classification of action. The TGR approach is demonstrated using "motion history images" that are then recognized via a mixture-of-Gaussians classifier. The system was tested in recognizing various dynamic human outdoor activities: running, walking, roller blading, and cycling. Experiments with real and synthetic data sets are used to evaluate stability of the trajectory estimator with respect to noise.

Differentiated Predictive Fair Service for TCP Flows

The majority of the traffic (bytes) flowing over the Internet today have been attributed to the Transmission Control Protocol (TCP). This strong presence of TCP has recently spurred further investigations into its congestion avoidance mechanism and its effect on the performance of short and long data transfers. At the same time, the rising interest in enhancing Internet services while keeping the implementation cost low has led to several service-differentiation proposals. In such service-differentiation architectures, much of the complexity is placed only in access routers, which classify and mark packets from different flows. Core routers can then allocate enough resources to each class of packets so as to satisfy delivery requirements, such as predictable (consistent) and fair service. In this paper, we investigate the interaction among short and long TCP flows, and how TCP service can be improved by employing a low-cost service-differentiation scheme. Through control-theoretic arguments and extensive simulations, we show the utility of isolating TCP flows into two classes based on their lifetime/size, namely one class of short flows and another of long flows. With such class-based isolation, short and long TCP flows have separate service queues at routers. This protects each class of flows from the other as they possess different characteristics, such as burstiness of arrivals/departures and congestion/sending window dynamics. We show the benefits of isolation, in terms of better predictability and fairness, over traditional shared queueing systems with both tail-drop and Random-Early-Drop (RED) packet dropping policies. The proposed class-based isolation of TCP flows has several advantages: (1) the implementation cost is low since it only requires core routers to maintain per-class (rather than per-flow) state; (2) it promises to be an effective traffic engineering tool for improved predictability and fairness for both short and long TCP flows; and (3) stringent delay requirements of short interactive transfers can be met by increasing the amount of resources allocated to the class of short flows.

An Efficient User-Level Shared Memory Mechanism for Application-Specific Extensions

This paper focuses on an efficient user-level method for the deployment of application-specific extensions, using commodity operating systems and hardware. A sandboxing technique is described that supports multiple extensions within a shared virtual address space. Applications can register sandboxed code with the system, so that it may be executed in the context of any process. Such code may be used to implement generic routines and handlers for a class of applications, or system service extensions that complement the functionality of the core kernel. Using our approach, application-specific extensions can be written like conventional user-level code, utilizing libraries and system calls, with the advantage that they may be executed without the traditional costs of scheduling and context-switching between process-level protection domains. No special hardware support such as segmentation or tagged translation look-aside buffers (TLBs) is required. Instead, our ``user-level sandboxing'' mechanism requires only paged-based virtual memory support, given that sandboxed extensions are either written by a trusted source or are guaranteed to be memory-safe (e.g., using type-safe languages). Using a fast method of upcalls, we show how our mechanism provides significant performance improvements over traditional methods of invoking user-level services. As an application of our approach, we have implemented a user-level network subsystem that avoids data copying via the kernel and, in many cases, yields far greater network throughput than kernel-level approaches.

A novel activation mechanism for Clostridial bacteriophage endolysins

Bacteriophage-encoded endolysins are produced at the end of the phage lytic cycle for the degradation of the host bacterial cell. Endolysins offer the potential as alternatives to antibiotics as biocontrol agents or therapeutics. The lytic mechanisms of three bacteriophage endolysins that target Clostridium species living under different conditions were investigated. For these endolysins a trigger and release mechanism is proposed for their activation. During host lysis, holin lesion formation suddenly permeabilises the membrane which exposes the cytosol-sequestered endolysins to a sudden environmental shock. This shock is suggested to trigger a conformational switch of the endolysins between two distinct dimer states. The switch between dimer states is proposed to activate a novel autocleavage mechanism that cleaves the linker connecting the N-terminal catalytic domain and the C-terminal domain to release the catalytic domain for more efficient digestion of the bacterial cell wall. Crystal structures of cleaved fragments of CD27L and CTP1L were previously obtained. In these structures cleavage occurs at the stem of the linker connected to the C-terminal domain. Despite a sequence identity of only 22% between 81 residues of the C-terminal domains of CD27L and CTP1L, they represent a novel fold that is identified in a number of different lysins. Within the crystal structures the two distinct dimerization modes are represented: the elongated head‐on dimer and the side-by‐side dimer. Introducing mutations that inhibit either of the dimerization states caused a decrease in the efficiency of both the autocleavage mechanism and the lytic activity of the endolysins. The two dimer states were validated for the full-length endolysins in solution by using right angle light scattering, small angle X‐ray scattering and cross-linking experiments. Overall, the data represents a new type of regulation governed by the C-terminal domains that is used to activate these endolysins once they enter the bacterial cell wall.

Emi2-mediated inhibition of E2-substrate ubiquitin transfer by the anaphase-promoting complex/cyclosome through a D-box-independent mechanism.

Vertebrate eggs are arrested at Metaphase II by Emi2, the meiotic anaphase-promoting complex/cyclosome (APC/C) inhibitor. Although the importance of Emi2 during oocyte maturation has been widely recognized and its regulation extensively studied, its mechanism of action remained elusive. Many APC/C inhibitors have been reported to act as pseudosubstrates, inhibiting the APC/C by preventing substrate binding. Here we show that a previously identified zinc-binding region is critical for the function of Emi2, whereas the D-box is largely dispensable. We further demonstrate that instead of acting through a "pseudosubstrate" mechanism as previously hypothesized, Emi2 can inhibit Cdc20-dependent activation of the APC/C substoichiometrically, blocking ubiquitin transfer from the ubiquitin-charged E2 to the substrate. These findings provide a novel mechanism of APC/C inhibition wherein the final step of ubiquitin transfer is targeted and raise the interesting possibility that APC/C is inhibited by Emi2 in a catalytic manner.

Osmolyte-induced folding of an intrinsically disordered protein: folding mechanism in the absence of ligand.

Understanding the interconversion between thermodynamically distinguishable states present in a protein folding pathway provides not only the kinetics and energetics of protein folding but also insights into the functional roles of these states in biological systems. The protein component of the bacterial RNase P holoenzyme from Bacillus subtilis (P protein) was previously shown to be unfolded in the absence of its cognate RNA or other anionic ligands. P protein was used in this study as a model system to explore general features of intrinsically disordered protein (IDP) folding mechanisms. The use of trimethylamine N-oxide (TMAO), an osmolyte that stabilizes the unliganded folded form of the protein, enabled us to study the folding process of P protein in the absence of ligand. Transient stopped-flow kinetic traces at various final TMAO concentrations exhibited multiphasic kinetics. Equilibrium "cotitration" experiments were performed using both TMAO and urea during the titration to produce a urea-TMAO titration surface of P protein. Both kinetic and equilibrium studies show evidence of a previously undetected intermediate state in the P protein folding process. The intermediate state is significantly populated, and the folding rate constants are relatively slow compared to those of intrinsically folded proteins similar in size and topology. The experiments and analysis described serve as a useful example for mechanistic folding studies of other IDPs.