Correlation between extent of osteolytic damage and metastatic burden of human breast cancer metastasis in nude mice : real-time PCR quantitation

Orthotopic or intracardiac injection of human breast cancer cell lines into immunocompromised mice allows study of the molecular basis of breast cancer metastasis. We have established a quantitative real-time PCR approach to analyze metastatic spread of human breast cancer cells inoculated into nude mice via these routes. We employed MDA-MB-231 human breast cancer cells genetically tagged with a bacterial β-galactosidase (Lac-Z) retroviral vector, enabling their detection by TaqMan® real-time PCR. PCR detection was linear, specific, more sensitive than conventional PCR, and could be used to directly quantitate metastatic burden in bone and soft organs. Attesting to the sensitivity and specificity of the PCR detection strategy, as few as several hundred metastatic MDA-MB-231 cells were detectable in 100 μm segments of paraffin-embedded lung tissue, and only in samples adjacent to sections that scored positive by histological detection. Moreover, the measured real-time PCR metastatic burden in the bone environment (mouse hind-limbs, n = 48) displayed a high correlation to the degree of osteolytic damage observed by high resolution X-ray analysis (r2 = 0.972). Such a direct linear relationship to tumor burden and bone damage substantiates the so-called 'vicious cycle' hypothesis in which metastatic tumor cells promote the release of factors from the bone which continue to stimulate the tumor cells. The technique provides a useful tool for molecular and cellular analysis of human breast cancer metastasis to bone and soft organs, can easily be extended to other cell/marker/organ systems, and should also find application in preclinical assessment of anti-metastatic modalities.

Methodology for developing real-time motorway traffic risk identification models using individual-vehicle data

Most of existing motorway traffic safety studies using disaggregate traffic flow data aim at developing models for identifying real-time traffic risks by comparing pre-crash and non-crash conditions. One of serious shortcomings in those studies is that non-crash conditions are arbitrarily selected and hence, not representative, i.e. selected non-crash data might not be the right data comparable with pre-crash data; the non-crash/pre-crash ratio is arbitrarily decided and neglects the abundance of non-crash over pre-crash conditions; etc. Here, we present a methodology for developing a real-time MotorwaY Traffic Risk Identification Model (MyTRIM) using individual vehicle data, meteorological data, and crash data. Non-crash data are clustered into groups called traffic regimes. Thereafter, pre-crash data are classified into regimes to match with relevant non-crash data. Among totally eight traffic regimes obtained, four highly risky regimes were identified; three regime-based Risk Identification Models (RIM) with sufficient pre-crash data were developed. MyTRIM memorizes the latest risk evolution identified by RIM to predict near future risks. Traffic practitioners can decide MyTRIM’s memory size based on the trade-off between detection and false alarm rates. Decreasing the memory size from 5 to 1 precipitates the increase of detection rate from 65.0% to 100.0% and of false alarm rate from 0.21% to 3.68%. Moreover, critical factors in differentiating pre-crash and non-crash conditions are recognized and usable for developing preventive measures. MyTRIM can be used by practitioners in real-time as an independent tool to make online decision or integrated with existing traffic management systems.

Towards real-time understanding of processes in pharmaceutical powder technology

There is a need for better understanding of the processes and new ideas to develop traditional pharmaceutical powder manufacturing procedures. Process analytical technology (PAT) has been developed to improve understanding of the processes and establish methods to monitor and control processes. The interest is in maintaining and even improving the whole manufacturing process and the final products at real-time. Process understanding can be a foundation for innovation and continuous improvement in pharmaceutical development and manufacturing. New methods are craved for to increase the quality and safety of the final products faster and more efficiently than ever before. The real-time process monitoring demands tools, which enable fast and noninvasive measurements with sufficient accuracy. Traditional quality control methods have been laborious and time consuming and they are performed off line i.e. the analysis has been removed from process area. Vibrational spectroscopic methods are responding this challenge and their utilisation have increased a lot during the past few years. In addition, other methods such as colour analysis can be utilised in noninvasive real-time process monitoring. In this study three pharmaceutical processes were investigated: drying, mixing and tabletting. In addition tablet properties were evaluated. Real-time monitoring was performed with NIR and Raman spectroscopies, colour analysis, particle size analysis and compression data during tabletting was evaluated using mathematical modelling. These methods were suitable for real-time monitoring of pharmaceutical unit operations and increase the knowledge of the critical parameters in the processes and the phenomena occurring during operations. They can improve our process understanding and therefore, finally, enhance the quality of final products.

Genetic variability of Tomato spotted wilt virus in Australia and validation of real time RT-PCR for its detection in single and bulked leaf samples

The potential for large-scale use of a sensitive real time reverse transcription polymerase chain reaction (RT-PCR) assay was evaluated for the detection of Tomato spotted wilt virus (TSWV) in single and bulked leaf samples by comparing its sensitivity with that of DAS-ELISA. Using total RNA extracted with RNeasy® or leaf soak methods, real time RT-PCR detected TSWV in all infected samples collected from 16 horticultural crop species (including flowers, herbs and vegetables), two arable crop species, and four weed species by both assays. In samples in which DAS-ELISA had previously detected TSWV, real time RT-PCR was effective at detecting it in leaf tissues of all 22 plant species tested at a wide range of concentrations. Bulk samples required more robust and extensive extraction methods with real time RT-PCR, but it generally detected one infected sample in 1000 uninfected ones. By contrast, ELISA was less sensitive when used to test bulked samples, once detecting up to 1 infected in 800 samples with pepper but never detecting more than 1 infected in 200 samples in tomato and lettuce. It was also less reliable than real time RT-PCR when used to test samples from parts of the leaf where the virus concentration was low. The genetic variability among Australian isolates of TSWV was small. Direct sequencing of a 587 bp region of the nucleoprotein gene (S RNA) of 29 isolates from diverse crops and geographical locations yielded a maximum of only 4.3% nucleotide sequence difference. Phylogenetic analysis revealed no obvious groupings of isolates according to geographic origin or host species. TSWV isolates, that break TSWV resistance genes in tomato or pepper did not differ significantly in the N gene region studied, indicating that a different region of the virus genome is responsible for this trait.

Detection of equine herpesvirus type 1 using a real-time polymerase chain reaction.

Equid herpesvirus 1 (EHV1) is a major disease of equids worldwide causing considerable losses to the horse industry. A variety of techniques, including PCR have been used to diagnose EHV1. Some of these PCRs were used in combination with other techniques such as restriction enzyme analysis (REA) or hybridisation, making them cumbersome for routine diagnostic testing and increasing the chances of cross-contamination. Furthermore, they involve the use of suspected carcinogens such as ethidium bromide and ultraviolet light. In this paper, we describe a real-time PCR, which uses minor groove-binding probe (MGB) technology for the diagnosis of EHV1. This technique does not require post-PCR manipulations thereby reducing the risk of cross-contamination. Most importantly, the technique is specific; it was able to differentiate EHV1 from the closely related member of the Alphaherpesvirinae, equid herpesvirus 4 (EHV4). It was not reactive with common opportunistic pathogens such as Escherichia coli, Klebsiella oxytoca, Pseudomonas aeruginosa and Enterobacter agglomerans often involved in abortion. Similarly, it did not react with equine pathogens such as Streptococcus equi, Streptococcus equisimilis, Streptococcus zooepidemicus, Taylorella equigenitalis and Rhodococcus equi, which also cause abortion. The results obtained with this technique agreed with results from published PCR methods. The assay was sensitive enough to detect EHV1 sequences in paraffin-embedded tissues and clinical samples. When compared to virus isolation, the test was more sensitive. This test will be useful for the routine diagnosis of EHV1 based on its specificity, sensitivity, ease of performance and rapidity.

Real-time wide-area loading margin sensitivity (WALMS) in power systems

Loading margin sensitivity (LMS) has been widely used in applications in the realm of voltage stability assessment and control. Typically, LMS is derived based on system equilibrium equations near bifurcation and therefore requires full detailed system model and significant computation effort. Availability of phasor measurement units (PMUs) due to the recent development of wide-area monitoring system (WAMS) provides an alternative computation-friendly approach for calculating LMS. With such motivation, this work proposes measurement-based wide-area loading margin sensitivity (WALMS) in bulk power systems. The proposed sensitivity, with its simplicity, has great potential to be embedded in real-time applications. Moreover, the calculation of the WALMS is not limited to low voltage near bifurcation point. A case study on IEEE 39-bus system verifies the proposed sensitivity. Finally, a voltage control scenario demonstrates the potential application of the WALMS.

S-nets: A Petri net based model for performance evaluation of real-time scheduling algorithms

A new class of nets, called S-nets, is introduced for the performance analysis of scheduling algorithms used in real-time systems Deterministic timed Petri nets do not adequately model the scheduling of resources encountered in real-time systems, and need to be augmented with resource places and signal places, and a scheduler block, to facilitate the modeling of scheduling algorithms. The tokens are colored, and the transition firing rules are suitably modified. Further, the concept of transition folding is used, to get intuitively simple models of multiframe real-time systems. Two generic performance measures, called �load index� and �balance index,� which characterize the resource utilization and the uniformity of workload distribution, respectively, are defined. The utility of S-nets for evaluating heuristic-based scheduling schemes is illustrated by considering three heuristics for real-time scheduling. S-nets are useful in tuning the hardware configuration and the underlying scheduling policy, so that the system utilization is maximized, and the workload distribution among the computing resources is balanced.

A Real-time clustering system for spatio-temporal signals from network of neurons

Over past few years, the studies of cultured neuronal networks have opened up avenues for understanding the ion channels, receptor molecules, and synaptic plasticity that may form the basis of learning and memory. The hippocampal neurons from rats are dissociated and cultured on a surface containing a grid of 64 electrodes. The signals from these 64 electrodes are acquired using a fast data acquisition system MED64 (Alpha MED Sciences, Japan) at a sampling rate of 20 K samples with a precision of 16-bits per sample. A few minutes of acquired data runs in to a few hundreds of Mega Bytes. The data processing for the neural analysis is highly compute-intensive because the volume of data is huge. The major processing requirements are noise removal, pattern recovery, pattern matching, clustering and so on. In order to interface a neuronal colony to a physical world, these computations need to be performed in real-time. A single processor such as a desk top computer may not be adequate to meet this computational requirements. Parallel computing is a method used to satisfy the real-time computational requirements of a neuronal system that interacts with an external world while increasing the flexibility and scalability of the application. In this work, we developed a parallel neuronal system using a multi-node Digital Signal processing system. With 8 processors, the system is able to compute and map incoming signals segmented over a period of 200 ms in to an action in a trained cluster system in real time.

Fast Robust Pattern Classification Algorithms for Real Time Neuro-Motor Prosthetic Applications

In this paper, we give a brief review of pattern classification algorithms based on discriminant analysis. We then apply these algorithms to classify movement direction based on multivariate local field potentials recorded from a microelectrode array in the primary motor cortex of a monkey performing a reaching task. We obtain prediction accuracies between 55% and 90% using different methods which are significantly above the chance level of 12.5%.

Real-time monitoring of critical nodes with minimal number of Phasor Measurement Units

This paper presents a method for placement of Phasor Measurement Units, ensuring the monitoring of vulnerable buses which are obtained based on transient stability analysis of the overall system. Real-time monitoring of phase angles across different nodes, which indicates the proximity to instability, the very purpose will be well defined if the PMUs are placed at buses which are more vulnerable. The issue is to identify the key buses where the PMUs should be placed when the transient stability prediction is taken into account considering various disturbances. Integer Linear Programming technique with equality and inequality constraints is used to find out the optimal placement set with key buses identified from transient stability analysis. Results on IEEE-14 bus system are presented to illustrate the proposed approach.

Ship-Mounted Real-Time Surface Observational System on board Indian Vessels for Validation and Refinement of Model Forcing Fields

A network of ship-mounted real-time Automatic Weather Stations integrated with Indian geosynchronous satellites Indian National Satellites (INSATs)] 3A and 3C, named Indian National Centre for Ocean Information Services Real-Time Automatic Weather Stations (I-RAWS), is established. The purpose of I-RAWS is to measure the surface meteorological-ocean parameters and transmit the data in real time in order to validate and refine the forcing parameters (obtained from different meteorological agencies) of the Indian Ocean Forecasting System (INDOFOS). Preliminary validation and intercomparison of analyzed products obtained from the National Centre for Medium Range Weather Forecasting and the European Centre for Medium-Range Weather Forecasts using the data collected from I-RAWS were carried out. This I-RAWS was mounted on board oceanographic research vessel Sagar Nidhi during a cruise across three oceanic regimes, namely, the tropical Indian Ocean, the extratropical Indian Ocean, and the Southern Ocean. The results obtained from such a validation and intercomparison, and its implications with special reference to the usage of atmospheric model data for forcing ocean model, are discussed in detail. It is noticed that the performance of analysis products from both atmospheric models is similar and good; however, European Centre for Medium-Range Weather Forecasts air temperature over the extratropical Indian Ocean and wind speed in the Southern Ocean are marginally better.

Real-Time Stress Measurements in Germanium Thin Film Electrodes during Electrochemical Lithiation/Delithiation Cycling

An in situ study of stress evolution and mechanical behavior of germanium as a lithium-ion battery electrode material is presented. Thin films of germanium are cycled in a half-cell configuration with lithium metal foil as counter/reference electrode, with 1M LiPF6 in ethylene carbonate, diethyl carbonate, dimethyl carbonate solution (1:1:1, wt%) as electrolyte. Real-time stress evolution in the germanium thin-film electrodes during electrochemical lithiation/delithiation is measured by monitoring the substrate curvature using the multi-beam optical sensing method. Upon lithiation a-Ge undergoes extensive plastic deformation, with a peak compressive stress reaching as high as -0.76 +/- 0.05 GPa (mean +/- standard deviation). The compressive stress decreases with lithium concentration reaching a value of approximately -0.3 GPa at the end of lithiation. Upon delithiation the stress quickly became tensile and follows a trend that mirrors the behavior on compressive side; the average peak tensile stress of the lithiated Ge samples was approximately 0.83 GPa. The peak tensile stress data along with the SEM analysis was used to estimate a lower bound fracture resistance of lithiated Ge, which is approximately 5.3 J/m(2). It was also observed that the lithiated Ge is rate sensitive, i.e., stress depends on how fast or slow the charging is carried out. (C) The Author(s) 2015. Published by ECS. This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 License (CC BY, http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse of the work in any medium, provided the original work is properly cited. All rights reserved.

Real-Time Measurement on Deformation Fields of Notched Samples Under Impact Tension

In this paper, a real-time and in situ optical measuring system is reported to observe high-velocity deformations of samples subjected to impact loading. The system consists of a high-speed camera, a He-Ne laser, a frame grabber, a synchronization device and analysis software based on digital correlation theory. The optical system has been adapted to investigate the dynamic deformation field and its evolution in notched samples loaded by an split Hopkinson tension bar, with a resolution of 50 pin and an accuracy of 0.5 mum. Results obtained in experiments are discussed and compared with numerical simulations. It is shown that the measuring system is effective and valid.

Sliding mode position control for real-time control of induction motors

A sliding mode position control for high-performance real-time applications of induction motors in developed in this work. The design also incorporates a simple flux estimator in order to avoid the flux sensors. Then, the proposed control scheme presents a low computational cost and therefore can be implemented easily in a real-time applications using a low cost DSP-processor. The stability analysis of the controller under parameter uncertainties and load disturbances in provided using Lyapunov stability theory. Finally, simulated and experimental results show that the proposed controller with the proposed observer provides a good trajectory tracking and that this scheme is robust with respect to plant parameter variations and external load disturbances.

Investigation of hypervelocity impact phenomena using real-time concurrent diagnostics

Hypervelocity impact of meteoroids and orbital debris poses a serious and growing threat to spacecraft. To study hypervelocity impact phenomena, a comprehensive ensemble of real-time concurrently operated diagnostics has been developed and implemented in the Small Particle Hypervelocity Impact Range (SPHIR) facility. This suite of simultaneously operated instrumentation provides multiple complementary measurements that facilitate the characterization of many impact phenomena in a single experiment. The investigation of hypervelocity impact phenomena described in this work focuses on normal impacts of 1.8 mm nylon 6/6 cylinder projectiles and variable thickness aluminum targets. The SPHIR facility two-stage light-gas gun is capable of routinely launching 5.5 mg nylon impactors to speeds of 5 to 7 km/s. Refinement of legacy SPHIR operation procedures and the investigation of first-stage pressure have improved the velocity performance of the facility, resulting in an increase in average impact velocity of at least 0.57 km/s. Results for the perforation area indicate the considered range of target thicknesses represent multiple regimes describing the non-monotonic scaling of target perforation with decreasing target thickness. The laser side-lighting (LSL) system has been developed to provide ultra-high-speed shadowgraph images of the impact event. This novel optical technique is demonstrated to characterize the propagation velocity and two-dimensional optical density of impact-generated debris clouds. Additionally, a debris capture system is located behind the target during every experiment to provide complementary information regarding the trajectory distribution and penetration depth of individual debris particles. The utilization of a coherent, collimated illumination source in the LSL system facilitates the simultaneous measurement of impact phenomena with near-IR and UV-vis spectrograph systems. Comparison of LSL images to concurrent IR results indicates two distinctly different phenomena. A high-speed, pressure-dependent IR-emitting cloud is observed in experiments to expand at velocities much higher than the debris and ejecta phenomena observed using the LSL system. In double-plate target configurations, this phenomena is observed to interact with the rear-wall several micro-seconds before the subsequent arrival of the debris cloud. Additionally, dimensional analysis presented by Whitham for blast waves is shown to describe the pressure-dependent radial expansion of the observed IR-emitting phenomena. Although this work focuses on a single hypervelocity impact configuration, the diagnostic capabilities and techniques described can be used with a wide variety of impactors, materials, and geometries to investigate any number of engineering and scientific problems.