Plasmodium vivax: Reverse transcriptase real-time PCR for gametocyte detection and quantitation in clinical samples

The proportion of Plasmodium vivax-infected subjects that carry mature gametocytes, and thus are potentially infectious, remains poorly characterized in endemic settings. Here, we describe a quantitative reverse transcriptase (RI) real-time PCR (qRT-PCR) that targets transcripts of the mature gametocyte-specific pvs25 gene. We found mature gametocytes in 42 of 44 (95.4%) P. vivax infections diagnosed during an ongoing cohort study in northwestern Brazil. SYBR green qRT-PCR was more sensitive than a conventional RT-PCR that targets the same gene. Molecular detection of gametocytes failed, however, when dried bloodspots were used for RNA isolation and complementary DNA synthesis. Estimating the number of pvs25 gene transcripts allowed for examining the potential infectiousness of gametocyte carriers in a quantitative way. We found that most (61.9%) gametocyte carriers were either asymptomatic or had subpatent parasitemias and would have been missed by routine malaria control strategies. However, potentially undiagnosed gametocyte carriers usually had low-density infections and contributed a small fraction (up to 4%) to the overall gametocyte burden in the community. Further studies are required to determine the relative contribution to malaria transmission of long-lasting but low-density gametocytemias in asymptomatic carriers that are left undiagnosed and untreated. (C) 2012 Elsevier Inc. All rights reserved.

Real time monitoring of DNA hybridization and replication using optical and acoustic biosensors

Während in den letzten Jahren zahlreiche Biosensoren zum spezifischen Nachweis von DNA entwickelt wurden, ist die Anwendung oberflächen-sensitiver Methoden auf enzymatische Reaktionen ein vergleichsweise neues Forschungsgebiet. Trotz der hohen Empfindlichkeit und der Möglichkeit zur Echtzeit-Beobachtung molekularer Prozesse, ist die Anwendung dieser Methoden nicht etabliert, da die Enzymaktivität durch die Nähe zur Oberfläche beeinträchtigt sein kann. Im Rahmen dieser Arbeit wurde die enzymatische Verlängerung immobilisierter DNA durch eine DNA Polymerase mit Hilfe von Oberflächenplasmonen-Fluoreszenzspektroskopie (SPFS) und einer Quarzkristall-Mikrowaage (QCM) untersucht. Die Synthese von DNA wurde im Fall der QCM als Massenzuwachs detektiert, der sich im Abfall der Resonanzfrequenz des Schwingquarzes und einem Anstieg seiner Dissipationsenergie ausdrückte. Die viskoelastischen Eigenschaften der DNA-Schichten wurden bestimmt, indem die erhaltenen Daten mit einem auf Voigt basierenden Modell ausgewertet wurden. SPFS nutzt das evaneszente elektromagnetische Feld, das mit Oberflächenplasmonen einhergeht, zur oberflächen-sensitiven Anregung von Chromophoren. Auf diese Weise wurde der Einbau von Farbstoff-markierten Nukleotiden in die entstehende DNA-Sequenz als Indikator für das Voranschreiten der Reaktion ausgenutzt. Beide Meßtechniken konnten erfolgreich zum Nachweis der DNA-Synthese herangezogen werden, wobei die katalytische Aktivität des Enzyms vergleichbar zu der in Lösung gemessenen war.

Development and validation of a real-time quantitative PCR assay for rapid identification of Bacillus anthracis in environmental samples

A real-time polymerase chain reaction (PCR) assay was developed for rapid identification of Bacillus anthracis in environmental samples. These samples often harbor Bacillus cereus bacteria closely related to B. anthracis, which may hinder its specific identification by resulting in false positive signals. The assay consists of two duplex real-time PCR: the first PCR allows amplification of a sequence specific of the B. cereus group (B. anthracis, B. cereus, Bacillus thuringiensis, Bacillus weihenstephanensis, Bacillus pseudomycoides, and Bacillus mycoides) within the phosphoenolpyruvate/sugar phosphotransferase system I gene and a B. anthracis specific single nucleotide polymorphism within the adenylosuccinate synthetase gene. The second real-time PCR assay targets the lethal factor gene from virulence plasmid pXO1 and the capsule synthesis gene from virulence plasmid pXO2. Specificity of the assay is enhanced by the use of minor groove binding probes and/or locked nucleic acids probes. The assay was validated on 304 bacterial strains including 37 B. anthracis, 67 B. cereus group, 54 strains of non-cereus group Bacillus, and 146 Gram-positive and Gram-negative bacteria strains. The assay was performed on various environmental samples spiked with B. anthracis or B. cereus spores. The assay allowed an accurate identification of B. anthracis in environmental samples. This study provides a rapid and reliable method for improving rapid identification of B. anthracis in field operational conditions.

A real-time video quality estimator for emerging wireless multimedia systems

Wireless Mesh Networks (WMNs) are increasingly deployed to enable thousands of users to share, create, and access live video streaming with different characteristics and content, such as video surveillance and football matches. In this context, there is a need for new mechanisms for assessing the quality level of videos because operators are seeking to control their delivery process and optimize their network resources, while increasing the user’s satisfaction. However, the development of in-service and non-intrusive Quality of Experience assessment schemes for real-time Internet videos with different complexity and motion levels, Group of Picture lengths, and characteristics, remains a significant challenge. To address this issue, this article proposes a non-intrusive parametric real-time video quality estimator, called MultiQoE that correlates wireless networks’ impairments, videos’ characteristics, and users’ perception into a predicted Mean Opinion Score. An instance of MultiQoE was implemented in WMNs and performance evaluation results demonstrate the efficiency and accuracy of MultiQoE in predicting the user’s perception of live video streaming services when compared to subjective, objective, and well-known parametric solutions.

Real time passenger information systems and quality of bus services

One of the main problems in urban areas is the steady growth in car ownership and traffic levels. Therefore, the challenge of sustainability is focused on a shift of the demand for mobility from cars to collective means of transport. For this end, buses are a key element of the public transport systems. In this respect Real Time Passenger Information (RTPI) systems help citizens change their travel behaviour towards more sustainable transport modes. This paper provides an assessment methodology which evaluates how RTPI systems improve the quality of bus services in two European cities, Madrid and Bremerhaven. In the case of Madrid, bus punctuality has increased by 3%. Regarding the travellers perception, Madrid raised its quality of service by 6% while Bremerhaven increased by 13%. On the other hand, the users ́ perception of Public Transport (PT) image increased by 14%.

High-quality real-time temporal segmentation tool for video editing software

The increasing use of video editing software has resulted in a necessity for faster and more efficient editing tools. Here, we propose a lightweight high-quality video indexing tool that is suitable for video editing software.

Temporal segmentation tool for high-quality real time video editing software

The increasing use of video editing software requires faster and more efficient editing tools. As a first step, these tools perform a temporal segmentation in shots that allows a later building of indexes describing the video content. Here, we propose a novel real-time high-quality shot detection strategy, suitable for the last generation of video editing software requiring both low computational cost and high quality results. While abrupt transitions are detected through a very fast pixel-based analysis, gradual transitions are obtained from an efficient edge-based analysis. Both analyses are reinforced with a motion analysis that helps to detect and discard false detections. This motion analysis is carried out exclusively over a reduced set of candidate transitions, thus maintaining the computational requirements demanded by new applications to fulfill user needs.

Hardware timestamping for image acquisition system based on FlexRIO and IEEE 1588 v2 Standard

Current fusion devices consist of multiple diagnostics and hundreds or even thousands of signals. This situation forces on multiple occasions to use distributed data acquisition systems as the best approach. In this type of distributed systems, one of the most important issues is the synchronization between signals, so that it is possible to have a temporal correlation as accurate as possible between the acquired samples of all channels. In last decades, many fusion devices use different types of video cameras to provide inside views of the vessel during operations and to monitor plasma behavior. The synchronization between each video frame and the rest of the different signals acquired from any other diagnostics is essential in order to know correctly the plasma evolution, since it is possible to analyze jointly all the information having accurate knowledge of their temporal correlation. The developed system described in this paper allows timestamping image frames in a real-time acquisition and processing system using 1588 clock distribution. The system has been implemented using FPGA based devices together with a 1588 synchronized timing card (see Fig.1). The solution is based on a previous system [1] that allows image acquisition and real-time image processing based on PXIe technology. This architecture is fully compatible with the ITER Fast Controllers [2] and offers integration with EPICS to control and monitor the entire system. However, this set-up is not able to timestamp the frames acquired since the frame grabber module does not present any type of timing input (IRIG-B, GPS, PTP). To solve this lack, an IEEE1588 PXI timing device its used to provide an accurate way to synchronize distributed data acquisition systems using the Precision Time Protocol (PTP) IEEE 1588 2008 standard. This local timing device can be connected to a master clock device for global synchronization. The timing device has a buffer timestamp for each PXI trigger line and requires tha- a software application assigns each frame the corresponding timestamp. The previous action is critical and cannot be achieved if the frame rate is high. To solve this problem, it has been designed a solution that distributes the clock from the IEEE 1588 timing card to all FlexRIO devices [3]. This solution uses two PXI trigger lines that provide the capacity to assign timestamps to every frame acquired and register events by hardware in a deterministic way. The system provides a solution for timestamping frames to synchronize them with the rest of the different signals.

P2P audio/video protocol with global positioning data in real time for mobile devices

In this paper, we propose an original method to geoposition an audio/video stream with multiple emitters that are at the same time receivers of the mixed signal. The obtained method is suitable when a list of positions within a known area is encoded with precision tailored to the visualization capabilities of the target device. Nevertheless, it is easily adaptable to new precision requirements, as well as parameterized data precision. This method extends a previously proposed protocol, without incurring in any performance penalty.

Improved Techniques for Video Compression and Communication

Thesis (Ph.D.)--University of Washington, 2016-06

Low-cost real-time gesture recognition

A major impediment to developing real-time computer vision systems has been the computational power and level of skill required to process video streams in real-time. This has meant that many researchers have either analysed video streams off-line or used expensive dedicated hardware acceleration techniques. Recent software and hardware developments have greatly eased the development burden of realtime image analysis leading to the development of portable systems using cheap PC hardware and software exploiting the Multimedia Extension (MMX) instruction set of the Intel Pentium chip. This paper describes the implementation of a computationally efficient computer vision system for recognizing hand gestures using efficient coding and MMX-acceleration to achieve real-time performance on low cost hardware.

Environmental scanning electron microscopy offers real-time morphological analysis of liposomes and niosomes

Liposomes have been imaged using a plethora of techniques. However, few of these methods offer the ability to study these systems in their natural hydrated state without the requirement of drying, staining, and fixation of the vesicles. However, the ability to image a liposome in its hydrated state is the ideal scenario for visualization of these dynamic lipid structures and environmental scanning electron microscopy (ESEM), with its ability to image wet systems without prior sample preparation, offers potential advantages to the above methods. In our studies, we have used ESEM to not only investigate the morphology of liposomes and niosomes but also to dynamically follow the changes in structure of lipid films and liposome suspensions as water condenses on to or evaporates from the sample. In particular, changes in liposome morphology were studied using ESEM in real time to investigate the resistance of liposomes to coalescence during dehydration thereby providing an alternative assay of liposome formulation and stability. Based on this protocol, we have also studied niosome-based systems and cationic liposome/DNA complexes. Copyright © Informa Healthcare.

Image enhancement of real-time television to benefit the visually impaired

Purpose: To examine the use of real-time, generic edge detection, image processing techniques to enhance the television viewing of the visually impaired. Design: Prospective, clinical experimental study. Method: One hundred and two sequential visually impaired (average age 73.8 ± 14.8 years; 59% female) in a single center optimized a dynamic television image with respect to edge detection filter (Prewitt, Sobel, or the two combined), color (red, green, blue, or white), and intensity (one to 15 times) of the overlaid edges. They then rated the original television footage compared with a black-and-white image displaying the edges detected and the original television image with the detected edges overlaid in the chosen color and at the intensity selected. Footage of news, an advertisement, and the end of program credits were subjectively assessed in a random order. Results: A Prewitt filter was preferred (44%) compared with the Sobel filter (27%) or a combination of the two (28%). Green and white were equally popular for displaying the detected edges (32%), with blue (22%) and red (14%) less so. The average preferred edge intensity was 3.5 ± 1.7 times. The image-enhanced television was significantly preferred to the original (P < .001), which in turn was preferred to viewing the detected edges alone (P < .001) for each of the footage clips. Preference was not dependent on the condition causing visual impairment. Seventy percent were definitely willing to buy a set-top box that could achieve these effects for a reasonable price. Conclusions: Simple generic edge detection image enhancement options can be performed on television in real-time and significantly enhance the viewing of the visually impaired. © 2007 Elsevier Inc. All rights reserved.

Quality assessment of video transmission in heterogeneous networks as applied to real-time telecare

This work looks into video quality assessment applied to the field of telecare and proposes an alternative metric to the more traditionally used PSNR based on the requirements of such an application. We show that the Pause Intensity metric introduced in [1] is also relevant and applicable to heterogeneous networks with a wireless last hop connected to a wired TCP backbone. We demonstrate through our emulation testbed that the impairments experienced in such a network architecture are dominated by continuity based impairments rather than artifacts, such as motion drift or blockiness. We also look into the implication of using Pause Intensity as a metric in terms of the overall video latency, which is potentially problematic should the video be sent and acted upon in real-time. We conclude that Pause Intensity may be used alongside the video characteristics which have been suggested as a measure of the overall video quality. © 2012 IEEE.