Real-time biosensor for the assessment of nanotoxicity and cancer electrotherapy

Knowledge of cell electronics has led to their integration to medicine either by physically interfacing electronic devices with biological systems or by using electronics for both detection and characterization of biological materials. In this dissertation, an electrical impedance sensor (EIS) was used to measure the electrode surface impedance changes from cell samples of human and environmental toxicity of nanoscale materials in 2D and 3D cell culture models. The impedimetric response of human lung fibroblasts and rainbow trout gill epithelial cells when exposed to various nanomaterials was tested to determine their kinetic effects towards the cells and to demonstrate the biosensor's ability to monitor nanotoxicity in real-time. Further, the EIS allowed rapid, real-time and multi-sample analysis creating a versatile, noninvasive tool that is able to provide quantitative information with respect to alteration in cellular function. We then extended the application of the unique capabilities of the EIS to do real-time analysis of cancer cell response to externally applied alternating electric fields at different intermediate frequencies and low-intensity. Decreases in the growth profiles of the ovarian and breast cancer cells were observed with the application of 200 and 100 kHz, respectively, indicating specific inhibitory effects on dividing cells in culture in contrast to the non-cancerous HUVECs and mammary epithelial cells. We then sought to enhance the effects of the electric field by altering the cancer cell's electronegative membrane properties with HER2 antibody functionalized nanoparticles. An Annexin V/EthD-III assay and zeta potential were performed to determine the cell death mechanism indicating apoptosis and a decrease in zeta potential with the incorporation of the nanoparticles. With more negatively charged HER2-AuNPs attached to the cancer cell membrane, the decrease in membrane potential would thus leave the cells more vulnerable to the detrimental effects of the applied electric field due to the decrease in surface charge. Therefore, by altering the cell membrane potential, one could possibly control the fate of the cell. This whole cell-based biosensor will enhance our understanding of the responsiveness of cancer cells to electric field therapy and demonstrate potential therapeutic opportunities for electric field therapy in the treatment of cancer.

Optimal kernel development for real-time communications

The purpose of this research is to develop an optimal kernel which would be used in a real-time engineering and communications system. Since the application is a real-time system, relevant real-time issues are studied in conjunction with kernel related issues. The emphasis of the research is the development of a kernel which would not only adhere to the criteria of a real-time environment, namely determinism and performance, but also provide the flexibility and portability associated with non-real-time environments. The essence of the research is to study how the features found in non-real-time systems could be applied to the real-time system in order to generate an optimal kernel which would provide flexibility and architecture independence while maintaining the performance needed by most of the engineering applications. Traditionally, development of real-time kernels has been done using assembly language. By utilizing the powerful constructs of the C language, a real-time kernel was developed which addressed the goals of flexibility and portability while still meeting the real-time criteria. The implementation of the kernel is carried out using the powerful 68010/20/30/40 microprocessor based systems.

The effects of mobile real-time information on rural passengers

The research described here is supported by the award made by the RCUK Digital Economy programme to the dot.rural Digital Economy Research Hub; award reference: EP/G066051/1/.

Video anomaly detection in real-time on a Power-Aware Heterogeneous Platform

FPGAs and GPUs are often used when real-time performance in video processing is required. An accelerated processor is chosen based on task-specific priorities (power consumption, processing time and detection accuracy), and this decision is normally made once at design time. All three characteristics are important, particularly in battery-powered systems. Here we propose a method for moving selection of processing platform from a single design-time choice to a continuous run time one.We implement Histogram of Oriented Gradients (HOG) detectors for cars and people and Mixture of Gaussians (MoG) motion detectors running across FPGA, GPU and CPU in a heterogeneous system. We use this to detect illegally parked vehicles in urban scenes. Power, time and accuracy information for each detector is characterised. An anomaly measure is assigned to each detected object based on its trajectory and location, when compared to learned contextual movement patterns. This drives processor and implementation selection, so that scenes with high behavioural anomalies are processed with faster but more power hungry implementations, but routine or static time periods are processed with power-optimised, less accurate, slower versions. Real-time performance is evaluated on video datasets including i-LIDS. Compared to power-optimised static selection, automatic dynamic implementation mapping is 10% more accurate but draws 12W extra power in our testbed desktop system.

Application of self-quenched JH consensus primers for real-time quantitative PCR of IGH gene to minimal residual disease evaluation in multiple myeloma.

Monitoring multiple myeloma patients for relapse requires sensitive methods to measure minimal residual disease and to establish a more precise prognosis. The present study aimed to standardize a real-time quantitative polymerase chain reaction (PCR) test for the IgH gene with a JH consensus self-quenched fluorescence reverse primer and a VDJH or DJH allele-specific sense primer (self-quenched PCR). This method was compared with allele-specific real-time quantitative PCR test for the IgH gene using a TaqMan probe and a JH consensus primer (TaqMan PCR). We studied nine multiple myeloma patients from the Spanish group treated with the MM2000 therapeutic protocol. Self-quenched PCR demonstrated sensitivity of >or=10(-4) or 16 genomes in most cases, efficiency was 1.71 to 2.14, and intra-assay and interassay reproducibilities were 1.18 and 0.75%, respectively. Sensitivity, efficiency, and residual disease detection were similar with both PCR methods. TaqMan PCR failed in one case because of a mutation in the JH primer binding site, and self-quenched PCR worked well in this case. In conclusion, self-quenched PCR is a sensitive and reproducible method for quantifying residual disease in multiple myeloma patients; it yields similar results to TaqMan PCR and may be more effective than the latter when somatic mutations are present in the JH intronic primer binding site.

Threat Analysis of BlackEnergy Malware for Synchrophasor based Real-time Control and Monitoring in Smart Grid

The BlackEnergy malware targeting critical infrastructures has a long history. It evolved over time from a simple DDoS platform to a quite sophisticated plug-in based malware. The plug-in architecture has a persistent malware core with easily installable attack specific modules for DDoS, spamming, info-stealing, remote access, boot-sector formatting etc. BlackEnergy has been involved in several high profile cyber physical attacks including the recent Ukraine power grid attack in December 2015. This paper investigates the evolution of BlackEnergy and its cyber attack capabilities. It presents a basic cyber attack model used by BlackEnergy for targeting industrial control systems. In particular, the paper analyzes cyber threats of BlackEnergy for synchrophasor based systems which are used for real-time control and monitoring functionalities in smart grid. Several BlackEnergy based attack scenarios have been investigated by exploiting the vulnerabilities in two widely used synchrophasor communication standards: (i) IEEE C37.118 and (ii) IEC 61850-90-5. Specifically, the paper addresses reconnaissance, DDoS, man-in-the-middle and replay/reflection attacks on IEEE C37.118 and IEC 61850-90-5. Further, the paper also investigates protection strategies for detection and prevention of BlackEnergy based cyber physical attacks.

A C80 DSP-based active vision system for real-time tracking

[EN]Active Vision Systems can be considered as dynamical systems which close the loop around artificial visual perception, controlling camera parameters, motion and also controlling processing to simplify, accelerate and do more robust visual perception. Research and Development in Active Vision Systems [Aloi87], [Bajc88] is a main area of interest in Computer Vision, mainly by its potential application in different scenarios where real-time performance is needed such as robot navigation, surveillance, visual inspection, among many others. Several systems have been developed during last years using robotic-heads for this purpose...

Recommendations for in-situ data Near Real Time Quality Control

With the construction of operational oceanography systems, the need for real-time has become more and more important. A lot of work had been done in the past, within National Data Centres (NODC) and International Oceanographic Data and Information Exchange (IODE) to standardise delayed mode quality control procedures. Concerning such quality control procedures applicable in real-time (within hours to a maximum of a week from acquisition), which means automatically, some recommendations were set up for physical parameters but mainly within projects without consolidation with other initiatives. During the past ten years the EuroGOOS community has been working on such procedures within international programs such as Argo, OceanSites or GOSUD, or within EC projects such as Mersea, MFSTEP, FerryBox, ECOOP, and MyOcean. In collaboration with the FP7 SeaDataNet project that is standardizing the delayed mode quality control procedures in NODCs, and MyOcean GMES FP7 project that is standardizing near real time quality control procedures for operational oceanography purposes, the DATA-MEQ working group decided to put together this document to summarize the recommendations for near real-time QC procedures that they judged mature enough to be advertised and recommended to EuroGOOS.

Dendritic Cells for Real-Time Anomaly Detection

Dendritic Cells (DCs) are innate immune system cells which have the power to activate or suppress the immune system. The behaviour of human DCs is abstracted to form an algorithm suitable for anomaly detection. We test this algorithm on the real-time problem of port scan detection. Our results show a significant difference in artificial DC behaviour for an outgoing portscan when compared to behaviour for normal processes.

RCOS: Real time context sharing across a fleet of smart mobile devices

Today, biodiversity is endangered by the currently applied intensive farming methods imposed on food producers by intermediate actors (e.g.: retailers). The lack of a direct communication technology between the food producer and the consumer creates dependency on the intermediate actors for both producers and the consumers. A tool allowing producers to directly and efficiently market produce that meets customer demands could greatly reduce the dependency enforced by intermediate actors. To this end, in this thesis, we propose, develop, implement and validate a Real Time Context Sharing (RCOS) system. RCOS takes advantage of the widely used publish/subscribe paradigm to exchange messages between producers and consumers, directly, according to their interest and context. Current systems follow a topic-based model or a content-based model. With RCOS, we propose a context-awareness approach into the matching process of publish/subscribe paradigm. Finally, as a proof of concept, we extend the Apache ActiveMQ Artemis software and create a client prototype. We evaluate our proof of concept for larger scale deployment.

INTEGRATED THRESHOLD-ACTIVATED FEEDBACK MICROSYSTEM FOR REAL-TIME CHARACTERIZATION, SENSING AND TREATMENT OF BACTERIAL BIOFILMS

Biofilms are the primary cause of clinical bacterial infections and are impervious to typical amounts of antibiotics, necessitating very high doses for treatment. Therefore, it is highly desirable to develop new alternate methods of treatment that can complement or replace existing approaches using significantly lower doses of antibiotics. Current standards for studying biofilms are based on end-point studies that are invasive and destroy the biofilm during characterization. This dissertation presents the development of a novel real-time sensing and treatment technology to aid in the non-invasive characterization, monitoring and treatment of bacterial biofilms. The technology is demonstrated through the use of a high-throughput bifurcation based microfluidic reactor that enables simulation of flow conditions similar to indwelling medical devices. The integrated microsystem developed in this work incorporates the advantages of previous in vitro platforms while attempting to overcome some of their limitations. Biofilm formation is extremely sensitive to various growth parameters that cause large variability in biofilms between repeated experiments. In this work we investigate the use of microfluidic bifurcations for the reduction in biofilm growth variance. The microfluidic flow cell designed here spatially sections a single biofilm into multiple channels using microfluidic flow bifurcation. Biofilms grown in the bifurcated device were evaluated and verified for reduced biofilm growth variance using standard techniques like confocal microscopy. This uniformity in biofilm growth allows for reliable comparison and evaluation of new treatments with integrated controls on a single device. Biofilm partitioning was demonstrated using the bifurcation device by exposing three of the four channels to various treatments. We studied a novel bacterial biofilm treatment independent of traditional antibiotics using only small molecule inhibitors of bacterial quorum sensing (analogs) in combination with low electric fields. Studies using the bifurcation-based microfluidic flow cell integrated with real-time transduction methods and macro-scale end-point testing of the combination treatment showed a significant decrease in biomass compared to the untreated controls and well-known treatments such as antibiotics. To understand the possible mechanism of action of electric field-based treatments, fundamental treatment efficacy studies focusing on the effect of the energy of the applied electrical signal were performed. It was shown that the total energy and not the type of the applied electrical signal affects the effectiveness of the treatment. The linear dependence of the treatment efficacy on the applied electrical energy was also demonstrated. The integrated bifurcation-based microfluidic platform is the first microsystem that enables biofilm growth with reduced variance, as well as continuous real-time threshold-activated feedback monitoring and treatment using low electric fields. The sensors detect biofilm growth by monitoring the change in impedance across the interdigitated electrodes. Using the measured impedance change and user inputs provided through a convenient and simple graphical interface, a custom-built MATLAB control module intelligently switches the system into and out of treatment mode. Using this self-governing microsystem, in situ biofilm treatment based on the principles of the bioelectric effect was demonstrated by exposing two of the channels of the integrated bifurcation device to low doses of antibiotics.

Real-time alert correlation with type graphs

The premise of automated alert correlation is to accept that false alerts from a low level intrusion detection system are inevitable and use attack models to explain the output in an understandable way. Several algorithms exist for this purpose which use attack graphs to model the ways in which attacks can be combined. These algorithms can be classified in to two broad categories namely scenario-graph approaches, which create an attack model starting from a vulnerability assessment and type-graph approaches which rely on an abstract model of the relations between attack types. Some research in to improving the efficiency of type-graph correlation has been carried out but this research has ignored the hypothesizing of missing alerts. Our work is to present a novel type-graph algorithm which unifies correlation and hypothesizing in to a single operation. Our experimental results indicate that the approach is extremely efficient in the face of intensive alerts and produces compact output graphs comparable to other techniques.

Real-time Biosensor for the Assessment of Nanotoxicity and Cancer Electrotherapy

Knowledge of cell electronics has led to their integration to medicine either by physically interfacing electronic devices with biological systems or by using electronics for both detection and characterization of biological materials. In this dissertation, an electrical impedance sensor (EIS) was used to measure the electrode surface impedance changes from cell samples of human and environmental toxicity of nanoscale materials in 2D and 3D cell culture models. The impedimetric response of human lung fibroblasts and rainbow trout gill epithelial cells when exposed to various nanomaterials was tested to determine their kinetic effects towards the cells and to demonstrate the biosensor’s ability to monitor nanotoxicity in real-time. Further, the EIS allowed rapid, real-time and multi-sample analysis creating a versatile, noninvasive tool that is able to provide quantitative information with respect to alteration in cellular function. We then extended the application of the unique capabilities of the EIS to do real-time analysis of cancer cell response to externally applied alternating electric fields at different intermediate frequencies and low-intensity. Decreases in the growth profiles of the ovarian and breast cancer cells were observed with the application of 200 and 100 kHz, respectively, indicating specific inhibitory effects on dividing cells in culture in contrast to the non-cancerous HUVECs and mammary epithelial cells. We then sought to enhance the effects of the electric field by altering the cancer cell’s electronegative membrane properties with HER2 antibody functionalized nanoparticles. An Annexin V/EthD-III assay and zeta potential were performed to determine the cell death mechanism indicating apoptosis and a decrease in zeta potential with the incorporation of the nanoparticles. With more negatively charged HER2-AuNPs attached to the cancer cell membrane, the decrease in membrane potential would thus leave the cells more vulnerable to the detrimental effects of the applied electric field due to the decrease in surface charge. Therefore, by altering the cell membrane potential, one could possibly control the fate of the cell. This whole cell-based biosensor will enhance our understanding of the responsiveness of cancer cells to electric field therapy and demonstrate potential therapeutic opportunities for electric field therapy in the treatment of cancer.

Introducing a pilot data collection model for real-time evaluation of data redundancy

In order to reduce serious health incidents, individuals with high risks need to be identified as early as possible so that effective intervention and preventive care can be provided. This requires regular and efficient assessments of risk within communities that are the first point of contacts for individuals. Clinical Decision Support Systems CDSSs have been developed to help with the task of risk assessment, however such systems and their underpinning classification models are tailored towards those with clinical expertise. Communities where regular risk assessments are required lack such expertise. This paper presents the continuation of GRiST research team efforts to disseminate clinical expertise to communities. Based on our earlier published findings, this paper introduces the framework and skeleton for a data collection and risk classification model that evaluates data redundancy in real-time, detects the risk-informative data and guides the risk assessors towards collecting those data. By doing so, it enables non-experts within the communities to conduct reliable Mental Health risk triage.