Mastering the canonical loop of serine protease inhibitors : enhancing potency by optimising the internal hydrogen bond network

Background Canonical serine protease inhibitors commonly bind to their targets through a rigid loop stabilised by an internal hydrogen bond network and disulfide bond(s). The smallest of these is sunflower trypsin inhibitor (SFTI-1), a potent and broad-range protease inhibitor. Recently, we re-engineered the contact β-sheet of SFTI-1 to produce a selective inhibitor of kallikrein-related peptidase 4 (KLK4), a protease associated with prostate cancer progression. However, modifications in the binding loop to achieve specificity may compromise structural rigidity and prevent re-engineered inhibitors from reaching optimal binding affinity. Methodology/Principal Findings In this study, the effect of amino acid substitutions on the internal hydrogen bonding network of SFTI were investigated using an in silico screen of inhibitor variants in complex with KLK4 or trypsin. Substitutions favouring internal hydrogen bond formation directly correlated with increased potency of inhibition in vitro. This produced a second generation inhibitor (SFTI-FCQR Asn14) which displayed both a 125-fold increased capacity to inhibit KLK4 (Ki = 0.0386±0.0060 nM) and enhanced selectivity over off-target serine proteases. Further, SFTI-FCQR Asn14 was stable in cell culture and bioavailable in mice when administered by intraperitoneal perfusion. Conclusion/Significance These findings highlight the importance of conserving structural rigidity of the binding loop in addition to optimising protease/inhibitor contacts when re-engineering canonical serine protease inhibitors.

Fusing loop detector and probe vehicle data to estimate travel time statistics on signalized urban networks

This article presents a methodology that integrates cumulative plots with probe vehicle data for estimation of travel time statistics (average, quartile) on urban networks. The integration reduces relative deviation among the cumulative plots so that the classical analytical procedure of defining the area between the plots as the total travel time can be applied. For quartile estimation, a slicing technique is proposed. The methodology is validated with real data from Lucerne, Switzerland and it is concluded that the travel time estimates from the proposed methodology are statistically equivalent to the observed values.

Algorithm for queue estimation with loop detector of time occupancy in off-ramps on signalized motorways

Long traffic queues on off-ramps significantly compromise the safety and throughput of motorways. Obtaining accurate queue information is crucial for countermeasure strategies. However, it is challenging to estimate traffic queues with locally installed inductive loop detectors. This paper deals with the problem of queue estimation with the interpretation of queuing dynamics and the corresponding time-occupancy distribution over motorway off-ramps. A novel algorithm for real-time queue estimation with two detectors is presented and discussed. Results derived from microscopic traffic simulation validated the effectiveness of the algorithm and revealed some of its useful features: (a) long and intermediate traffic queues could be accurately measured, (b) relatively simple detector input (i.e., time occupancy) was required, and (c) the estimation philosophy was independent with signal timing changes and provided the potential to cooperate with advanced strategies for signal control. Some issues concerning field implementation are also discussed.

Traffic queue estimation for metered motorway on-ramps through use of loop detector time occupancies

The primary objective of this study is to develop a robust queue estimation algorithm for motorway on-ramps. Real-time queue information is a vital input for dynamic queue management on metered on-ramps. Accurate and reliable queue information enables the management of on-ramp queue in an adaptive manner to the actual traffic queue size and thus minimises the adverse impacts of queue flush while increasing the benefit of ramp metering. The proposed algorithm is developed based on the Kalman filter framework. The fundamental conservation model is used to estimate the system state (queue size) with the flow-in and flow-out measurements. This projection results are updated with the measurement equation using the time occupancies from mid-link and link-entrance loop detectors. This study also proposes a novel single point correction method. This method resets the estimated system state to eliminate the counting errors that accumulate over time. In the performance evaluation, the proposed algorithm demonstrated accurate and reliable performances and consistently outperformed the benchmarked Single Occupancy Kalman filter (SOKF) method. The improvements over SOKF are 62% and 63% in average in terms of the estimation accuracy (MAE) and reliability (RMSE), respectively. The benefit of the innovative concepts of the algorithm is well justified by the improved estimation performance in congested ramp traffic conditions where long queues may significantly compromise the benchmark algorithm’s performance.

Studies in power hardware in the loop (PHIL) simulation using real-time digital simulator (RTDS)

In power hardware in the loop (PHIL) simulations, a real-time simulated power system is interfaced to a piece of hardware, usually called hardware under test (HuT). A PHIL test can be realized using several simulation tools. Among them Real Time Digital Simulator (RTDS) is an ideal tool to perform complex power system simulations in near real-time. Stable operation of the entire system, along with the accuracy of simulation results are the main concerns regarding a PHIL simulation. In this paper, a simulated power network on RTDS will be interfaced to HuT through a voltage source converter (VSC). Issues around stability and other interface problems are studied and a new method to stabilize some unstable PHIL cases is proposed. PHIL simulation results in PSCAD and RSCAD are presented.

Model-predictive control and closed-loop stability considerations for nonlinear plants described by local ARX-type models

In this paper, a model-predictive control (MPC) method is detailed for the control of nonlinear systems with stability considerations. It will be assumed that the plant is described by a local input/output ARX-type model, with the control potentially included in the premise variables, which enables the control of systems that are nonlinear in both the state and control input. Additionally, for the case of set point regulation, a suboptimal controller is derived which has the dual purpose of ensuring stability and enabling finite-iteration termination of the iterative procedure used to solve the nonlinear optimization problem that is used to determine the control signal.

Autonomous multisensor calibration and closed-loop fusion for SLAM

The vast majority of current robot mapping and navigation systems require specific well-characterized sensors that may require human-supervised calibration and are applicable only in one type of environment. Furthermore, if a sensor degrades in performance, either through damage to itself or changes in environmental conditions, the effect on the mapping system is usually catastrophic. In contrast, the natural world presents robust, reasonably well-characterized solutions to these problems. Using simple movement behaviors and neural learning mechanisms, rats calibrate their sensors for mapping and navigation in an incredibly diverse range of environments and then go on to adapt to sensor damage and changes in the environment over the course of their lifetimes. In this paper, we introduce similar movement-based autonomous calibration techniques that calibrate place recognition and self-motion processes as well as methods for online multisensor weighting and fusion. We present calibration and mapping results from multiple robot platforms and multisensory configurations in an office building, university campus, and forest. With moderate assumptions and almost no prior knowledge of the robot, sensor suite, or environment, the methods enable the bio-inspired RatSLAM system to generate topologically correct maps in the majority of experiments.

A phase-locked-loop design for the smooth operation of a hybrid microgrid

A microgrid contains both distributed generators (DGs) and loads and can be viewed by a controllable load by utilities. The DGs can be either inertial synchronous generators or non-inertial converter interfaced. Moreover, some of them can come online or go offline in plug and play fashion. The combination of these various types of operation makes the microgrid control a challenging task, especially when the microgrid operates in an autonomous mode. In this paper, a new phase locked loop (PLL) algorithm is proposed for smooth synchronization of plug and play DGs. A frequency droop for power sharing is used and a pseudo inertia has been introduced to non-inertial DGs in order to match their response with inertial DGs. The proposed strategy is validated through PSCAD simulation studies.

Distribution state estimation based on particle swarm and doubly loop mutant optimization (DLM-PSO)

This paper presents a novel algorithm based on particle swarm optimization (PSO) to estimate the states of electric distribution networks. In order to improve the performance, accuracy, convergence speed, and eliminate the stagnation effect of original PSO, a secondary PSO loop and mutation algorithm as well as stretching function is proposed. For accounting uncertainties of loads in distribution networks, pseudo-measurements is modeled as loads with the realistic errors. Simulation results on 6-bus radial and 34-bus IEEE test distribution networks show that the distribution state estimation based on proposed DLM-PSO presents lower estimation error and standard deviation in comparison with algorithms such as WLS, GA, HBMO, and original PSO.

Editorial : Loop

This time last year we proposed the theme of the 'loop' issue to the M/C collective because it sounded deeply cool, satisfying our poststructuralist posturings about reflexivity and representation, while also tapping into everyday cultural objects and practices. We expected that the 'loop' issue would generate some interesting and varied responses, and it did. We received submissions about music, visual art, language, child development, pop-cultural artefacts, mathematics and culture in general. These explorations of disparate fields seemed, however, to be tied together by a common thread: the concept of "generation" itself. Each article in the 'loop' issue describes a loop that does not simply repeat an original operation, but that through iteration creates new possibilities and new meanings...

Using performance-based evaluation to close the loop between biological and robotic navigation

In this paper we describe the benefits of a performance-based approach to modeling biological systems for use in robotics. Specifically, we describe the RatSLAM system, a computational model of the navigation processes thought to drive navigation in a part of the rodent brain called the hippocampus. Unlike typical computational modeling approaches, which focus on biological fidelity, RatSLAM’s development cycle has been driven primarily by performance evaluation on robots navigating in a wide variety of challenging, real world environments. We briefly describe three seminal results, two in robotics and one in biology. In addition, we present current research on brain-inspired learning algorithms with the aim of enabling a robot to autonomously learn how best to use its sensor suite to navigate, without requiring any specific knowledge of the robot, sensor types or environment characteristics. Our aim is to drive discussion on the merits of practical, performance-focused implementations of biological models in robotics.

Total energy shaping of a class of underactuated Port-Hamiltonian Systems using a new set of closed-loop potential shape variables

This paper proposes a method for designing set-point regulation controllers for a class of underactuated mechanical systems in Port-Hamiltonian System (PHS) form. A new set of potential shape variables in closed loop is proposed, which can replace the set of open loop shape variables-the configuration variables that appear in the kinetic energy. With this choice, the closed-loop potential energy contains free functions of the new variables. By expressing the regulation objective in terms of these new potential shape variables, the desired equilibrium can be assigned and there is freedom to reshape the potential energy to achieve performance whilst maintaining the PHS form in closed loop. This complements contemporary results in the literature, which preserve the open-loop shape variables. As a case study, we consider a robotic manipulator mounted on a flexible base and compensate for the motion of the base while positioning the end effector with respect to the ground reference. We compare the proposed control strategy with special cases that correspond to other energy shaping strategies previously proposed in the literature.

Quantitative stem-loop RT-PCR for detection of microRNAs

Plant microRNAs (miRNAs) are a class of endogenous small RNAs that are essential for plant development and survival. They arise from larger precursor RNAs with a characteristic hairpin structure and regulate gene activity by targeting mRNA transcripts for cleavage or translational repression. Efficient and reliable detection and quantification of miRNA expression has become an essential step in understanding their specific roles. The expression levels of miRNAs can vary dramatically between samples and they often escape detection by conventional technologies such as cloning, northern hybridization and microarray analysis. The stem-loop RT-PCR method described here is designed to detect and quantify mature miRNAs in a fast, specific, accurate and reliable manner. First, a miRNA-specific stem-loop RT primer is hybridized to the miRNA and then reverse transcribed. Next, the RT product is amplified and monitored in real time using a miRNA-specific forward primer and the universal reverse primer. This method enables miRNA expression profiling from as little as 10 pg of total RNA and is suitable for high-throughput miRNA expression analysis.

An arteriovenous loop in a protected space generates a permanent, highly vascular, tissue-engineered construct

A major obstacle to 3-dimensional tissue engineering is incorporation of a functional vascular supply to support the expanding new tissue. This is overcome in an in vivo intrinsic vascularization model where an arteriovenous loop (AVL) is placed in a noncollapsible space protected by a polycarbonate chamber. Vascular development and hypoxia were examined from 3 days to 112 days by vascular casting, morphometric, and morphological techniques to understand the model's vascular growth and remodeling parameters for tissue engineering purposes. At 3 days a fibrin exudate surrounded the AVL, providing a scaffold to migrating inflammatory, endothelial, and mesenchymal cells. Capillaries formed between 3 and 7 days. Hypoxia and cell proliferation were maximal at 7 days, followed by a peak in percent vascular volume at 10 days (23.20±3.14% compared with 3.59±2.68% at 3 days, P<0.001). Maximal apoptosis was observed at 112 days. The protected space and spontaneous microcirculatory development in this model suggest it would be applicable for in vivo tissue engineering. A temporal window in a period of intense angiogenesis at 7 to 10 days is optimal for exogenous cell seeding and survival in the chamber, potentially enabling specific tissue outcomes to be achieved.