Identification of reference genes for expression analysis by real-time quantitative PCR in drought-stressed soybean

O objetivo deste trabalho foi validar, pela técnica de PCR quantitativo em tempo real (RT-qPCR) genes para serem utilizados como referência em estudos de expressão gênica em soja, em ensaios de estresse hídrico. Foram avaliados quatro genes comumente utilizados em soja: Gmβ-actin, GmGAPDH, GmLectin e GmRNAr18S. O RNA total foi extraído de seis amostras: três amostras de raízes em sistema de hidroponia com diferentes intensidades de déficit hídrico (0, 25, 50, 75 e 100 minutos de estresse hídrico), e três amostras de folhas de plantas cultivadas em areia com diferentes umidades do solo (15, 5 e 2,5% de umidade gravimétrica). Os dados brutos do intervalo cycle threshold (Ct) foram analisados, e a eficiência de cada iniciador foi calculada para uma analise da Ct entre as diferentes amostras. A aplicação do programa GeNorm foi utilizada para a avaliação dos melhores genes de referência, de acordo com a estabilidade. O GmGAPDH foi o gene menos estável, com o maior valor médio de estabilidade de expressão (M), e os genes mais estáveis, com menor valor de M, foram o Gmβ-actin e GmRNAr18S, tanto nas amostras de raízes como nas de folhas. Estes genes podem ser usados em RT-qPCR como gens de referência para análises de expressão gênica.

Light-induced lens analysis in photorefractive crystals employing phase-shifting real-time holographic interferometry

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Development of an instrumentation system embedded on FPGA for real time measurement of mechanical vibrations in rotating machinery

The real-time monitoring of events in an industrial plant is vital, to monitor the actual conditions of operation of the machinery responsible for the manufacturing process. A predictive maintenance program includes condition monitoring of the rotating machinery, to anticipate possible conditions of failure. To increase the operational reliability it is thus necessary an efficient tool to analyze and monitor the equipments, in real-time, and enabling the detection of e.g. incipient faults in bearings. To fulfill these requirements some innovations have become frequent, namely the inclusion of vibration sensors or stator current sensors. These innovations enable the development of new design methodologies that take into account the ease of future modifications, upgrades, and replacement of the monitored machine, as well as expansion of the monitoring system. This paper presents the development, implementation and testing of an instrument for vibration monitoring, as a possible solution to embed in industrial environment. The digital control system is based on an FPGA, and its configuration with an open hardware design tool is described. Special focus is given to the area of fault detection in rolling bearings. © 2012 IEEE.

In situ real-time investigation of Organic Ultra-Thin-Film transistors: growth, electrical properties and biosensing applications

Organic electronics has grown enormously during the last decades driven by the encouraging results and the potentiality of these materials for allowing innovative applications, such as flexible-large-area displays, low-cost printable circuits, plastic solar cells and lab-on-a-chip devices. Moreover, their possible field of applications reaches from medicine, biotechnology, process control and environmental monitoring to defense and security requirements. However, a large number of questions regarding the mechanism of device operation remain unanswered. Along the most significant is the charge carrier transport in organic semiconductors, which is not yet well understood. Other example is the correlation between the morphology and the electrical response. Even if it is recognized that growth mode plays a crucial role into the performance of devices, it has not been exhaustively investigated. The main goal of this thesis was the finding of a correlation between growth modes, electrical properties and morphology in organic thin-film transistors (OTFTs). In order to study the thickness dependence of electrical performance in organic ultra-thin-film transistors, we have designed and developed a home-built experimental setup for performing real-time electrical monitoring and post-growth in situ electrical characterization techniques. We have grown pentacene TFTs under high vacuum conditions, varying systematically the deposition rate at a fixed room temperature. The drain source current IDS and the gate source current IGS were monitored in real-time; while a complete post-growth in situ electrical characterization was carried out. At the end, an ex situ morphological investigation was performed by using the atomic force microscope (AFM). In this work, we present the correlation for pentacene TFTs between growth conditions, Debye length and morphology (through the correlation length parameter). We have demonstrated that there is a layered charge carriers distribution, which is strongly dependent of the growth mode (i.e. rate deposition for a fixed temperature), leading to a variation of the conduction channel from 2 to 7 monolayers (MLs). We conciliate earlier reported results that were apparently contradictory. Our results made evident the necessity of reconsidering the concept of Debye length in a layered low-dimensional device. Additionally, we introduce by the first time a breakthrough technique. This technique makes evident the percolation of the first MLs on pentacene TFTs by monitoring the IGS in real-time, correlating morphological phenomena with the device electrical response. The present thesis is organized in the following five chapters. Chapter 1 makes an introduction to the organic electronics, illustrating the operation principle of TFTs. Chapter 2 presents the organic growth from theoretical and experimental points of view. The second part of this chapter presents the electrical characterization of OTFTs and the typical performance of pentacene devices is shown. In addition, we introduce a correcting technique for the reconstruction of measurements hampered by leakage current. In chapter 3, we describe in details the design and operation of our innovative home-built experimental setup for performing real-time and in situ electrical measurements. Some preliminary results and the breakthrough technique for correlating morphological and electrical changes are presented. Chapter 4 meets the most important results obtained in real-time and in situ conditions, which correlate growth conditions, electrical properties and morphology of pentacene TFTs. In chapter 5 we describe applicative experiments where the electrical performance of pentacene TFTs has been investigated in ambient conditions, in contact to water or aqueous solutions and, finally, in the detection of DNA concentration as label-free sensor, within the biosensing framework.

Dynamic distance analysis : new geometric data structures and algorithms for the real-time calculation of tolerance violating regions in the digital mockup process

In vielen Industriezweigen, zum Beispiel in der Automobilindustrie, werden Digitale Versuchsmodelle (Digital MockUps) eingesetzt, um die Konstruktion und die Funktion eines Produkts am virtuellen Prototypen zu überprüfen. Ein Anwendungsfall ist dabei die Überprüfung von Sicherheitsabständen einzelner Bauteile, die sogenannte Abstandsanalyse. Ingenieure ermitteln dabei für bestimmte Bauteile, ob diese in ihrer Ruhelage sowie während einer Bewegung einen vorgegeben Sicherheitsabstand zu den umgebenden Bauteilen einhalten. Unterschreiten Bauteile den Sicherheitsabstand, so muss deren Form oder Lage verändert werden. Dazu ist es wichtig, die Bereiche der Bauteile, welche den Sicherhabstand verletzen, genau zu kennen. rnrnIn dieser Arbeit präsentieren wir eine Lösung zur Echtzeitberechnung aller den Sicherheitsabstand unterschreitenden Bereiche zwischen zwei geometrischen Objekten. Die Objekte sind dabei jeweils als Menge von Primitiven (z.B. Dreiecken) gegeben. Für jeden Zeitpunkt, in dem eine Transformation auf eines der Objekte angewendet wird, berechnen wir die Menge aller den Sicherheitsabstand unterschreitenden Primitive und bezeichnen diese als die Menge aller toleranzverletzenden Primitive. Wir präsentieren in dieser Arbeit eine ganzheitliche Lösung, welche sich in die folgenden drei großen Themengebiete unterteilen lässt.rnrnIm ersten Teil dieser Arbeit untersuchen wir Algorithmen, die für zwei Dreiecke überprüfen, ob diese toleranzverletzend sind. Hierfür präsentieren wir verschiedene Ansätze für Dreiecks-Dreiecks Toleranztests und zeigen, dass spezielle Toleranztests deutlich performanter sind als bisher verwendete Abstandsberechnungen. Im Fokus unserer Arbeit steht dabei die Entwicklung eines neuartigen Toleranztests, welcher im Dualraum arbeitet. In all unseren Benchmarks zur Berechnung aller toleranzverletzenden Primitive beweist sich unser Ansatz im dualen Raum immer als der Performanteste.rnrnDer zweite Teil dieser Arbeit befasst sich mit Datenstrukturen und Algorithmen zur Echtzeitberechnung aller toleranzverletzenden Primitive zwischen zwei geometrischen Objekten. Wir entwickeln eine kombinierte Datenstruktur, die sich aus einer flachen hierarchischen Datenstruktur und mehreren Uniform Grids zusammensetzt. Um effiziente Laufzeiten zu gewährleisten ist es vor allem wichtig, den geforderten Sicherheitsabstand sinnvoll im Design der Datenstrukturen und der Anfragealgorithmen zu beachten. Wir präsentieren hierzu Lösungen, die die Menge der zu testenden Paare von Primitiven schnell bestimmen. Darüber hinaus entwickeln wir Strategien, wie Primitive als toleranzverletzend erkannt werden können, ohne einen aufwändigen Primitiv-Primitiv Toleranztest zu berechnen. In unseren Benchmarks zeigen wir, dass wir mit unseren Lösungen in der Lage sind, in Echtzeit alle toleranzverletzenden Primitive zwischen zwei komplexen geometrischen Objekten, bestehend aus jeweils vielen hunderttausend Primitiven, zu berechnen. rnrnIm dritten Teil präsentieren wir eine neuartige, speicheroptimierte Datenstruktur zur Verwaltung der Zellinhalte der zuvor verwendeten Uniform Grids. Wir bezeichnen diese Datenstruktur als Shrubs. Bisherige Ansätze zur Speicheroptimierung von Uniform Grids beziehen sich vor allem auf Hashing Methoden. Diese reduzieren aber nicht den Speicherverbrauch der Zellinhalte. In unserem Anwendungsfall haben benachbarte Zellen oft ähnliche Inhalte. Unser Ansatz ist in der Lage, den Speicherbedarf der Zellinhalte eines Uniform Grids, basierend auf den redundanten Zellinhalten, verlustlos auf ein fünftel der bisherigen Größe zu komprimieren und zur Laufzeit zu dekomprimieren.rnrnAbschießend zeigen wir, wie unsere Lösung zur Berechnung aller toleranzverletzenden Primitive Anwendung in der Praxis finden kann. Neben der reinen Abstandsanalyse zeigen wir Anwendungen für verschiedene Problemstellungen der Pfadplanung.

Real-time image processing for crop/weed discrimination in maize fields

This paper presents a computer vision system that successfully discriminates between weed patches and crop rows under uncontrolled lighting in real-time. The system consists of two independent subsystems, a fast image processing delivering results in real-time (Fast Image Processing, FIP), and a slower and more accurate processing (Robust Crop Row Detection, RCRD) that is used to correct the first subsystem's mistakes. This combination produces a system that achieves very good results under a wide variety of conditions. Tested on several maize videos taken of different fields and during different years, the system successfully detects an average of 95% of weeds and 80% of crops under different illumination, soil humidity and weed/crop growth conditions. Moreover, the system has been shown to produce acceptable results even under very difficult conditions, such as in the presence of dramatic sowing errors or abrupt camera movements. The computer vision system has been developed for integration into a treatment system because the ideal setup for any weed sprayer system would include a tool that could provide information on the weeds and crops present at each point in real-time, while the tractor mounting the spraying bar is moving

Real time monitoring of click chemistry self-healing in polymer composites

A photo-healable rubber composite based on effective and fast thiol-alkyne click chemistry as a selfhealing agent prestored in glass capillaries is reported. The click reaction and its effect on the mechanical properties of the composite are monitored in real time by dynamic mechanical analysis, showing that the successful bleeding of healing agents to the crack areas and the effective photoinitiated click reaction result in a 30% storage modulus increase after only 5 min of UV light exposure. X-ray tomography confirms capillary-driven bleeding of reactants to the damaged areas. The effect of storing the click chemistry reactants in separate capillaries is also studied, and results show the importance of stoichiometry in achieving a significant level of repair of the composite. No reactant degradation or premature chemical reaction is observed over time in samples stored in the absence of UV radiation; they are able to undergo the self-healing reaction even one month after preparation.

Real-time incidents detection in the highways of the future

Due to ever increasing transportation of people and goods, automatic traffic surveillance is becoming a key issue for both providing safety to road users and improving traffic control in an efficient way. In this paper, we propose a new system that, exploiting the capabilities that both computer vision and machine learning offer, is able to detect and track different types of real incidents on a highway. Specifically, it is able to accurately detect not only stopped vehicles, but also drivers and passengers leaving the stopped vehicle, and other pedestrians present in the roadway. Additionally, a theoretical approach for detecting vehicles which may leave the road in an unexpected way is also presented. The system works in real-time and it has been optimized for working outdoor, being thus appropriate for its deployment in a real-world environment like a highway. First experimental results on a dataset created with videos provided by two Spanish highway operators demonstrate the effectiveness of the proposed system and its robustness against noise and low-quality videos.

In-Motes EYE:a real time application for automobiles in wireless sensor networks

Wireless sensor networks have been identified as one of the key technologies for the 21st century. In order to overcome their limitations such as fault tolerance and conservation of energy, we propose a middleware solution, In-Motes. In-Motes stands as a fault tolerant platform for deploying and monitoring applications in real time offers a number of possibilities for the end user giving him in parallel the freedom to experiment with various parameters, in an effort the deployed applications to run in an energy efficient manner inside the network. The proposed scheme is evaluated through the In-Motes EYE application, aiming to test its merits under real time conditions. In-Motes EYE application which is an agent based real time In-Motes application developed for sensing acceleration variations in an environment. The application was tested in a prototype area, road alike, for a period of four months.

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.

Real-time high-resolution heterodyne-based measurements of spectral dynamics in fibre lasers

Conventional tools for measurement of laser spectra (e.g. optical spectrum analysers) capture data averaged over a considerable time period. However, the generation spectrum of many laser types may involve spectral dynamics whose relatively fast time scale is determined by their cavity round trip period, calling for instrumentation featuring both high temporal and spectral resolution. Such real-time spectral characterisation becomes particularly challenging if the laser pulses are long, or they have continuous or quasi-continuous wave radiation components. Here we combine optical heterodyning with a technique of spatiooral intensity measurements that allows the characterisation of such complex sources. Fast, round-trip-resolved spectral dynamics of cavity-based systems in real-time are obtained, with temporal resolution of one cavity round trip and frequency resolution defined by its inverse (85 ns and 24 MHz respectively are demonstrated). We also show how under certain conditions for quasi-continuous wave sources, the spectral resolution could be further increased by a factor of 100 by direct extraction of phase information from the heterodyned dynamics or by using double time scales within the spectrogram approach.

Westminster Serious Games Platform (wmin-SGP) a tool for real-time authoring of roleplay simulations for learning

The use of serious games in education and their pedagogical benefit is being widely recognized. However, effective integration of serious games in education depends on addressing two big challenges: the successful incorporation of motivation and engagement that can lead to learning; and the highly specialised skills associated with customised development to meet the required pedagogical objectives. This paper presents the Westminster Serious Games Platform (wmin-SGP) an authoring tool that allows educators/domain experts without games design and development technical skills to create bespoke roleplay simulations in three dimensional scenes featuring fully embodied virtual humans capable of verbal and non-verbal interaction with users fit for specific educational objectives. The paper presents the wmin-SGP system architecture and it evaluates its effectiveness in fulfilling its purpose via the implementation of two roleplay simulations, one for Politics and one for Law. In addition, it presents the results of two types of evaluation that address how successfully the wmin-SGP combines usability principles and game core drives based on the Octalysis gamification framework that lead to motivating games experiences. The evaluation results shows that the wmin-SGP: provides an intuitive environment and tools that support users without advanced technical skills to create in real-time bespoke roleplay simulations in advanced graphical interfaces; satisfies most of the usability principles; and provides balanced simulations based on the Octalysis framework core drives. The paper concludes with a discussion of future extension of this real time authoring tool and directions for further development of the Octalysis framework to address learning.

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.

Structuring real-time Object-Z specifications

This paper presents a means of structuring specifications in real-time Object-Z: an integration of Object-Z with the timed refinement calculus. Incremental modification of classes using inheritance and composition of classes to form multi-component systems are examined. Two approaches to the latter are considered: using Object-Z's notion of object instantiation and introducing a parallel composition operator similar to those found in process algebras. The parallel composition operator approach is both more concise and allows more general modelling of concurrency. Its incorporation into the existing semantics of real-time Object-Z is presented.

Real-time detection of pneumothorax using electrical impedance tomogyaphy

Objectives: Pneumothorax is a frequent complication during mechanical ventilation. Electrical impedance tomography (EIT) is a noninvasive tool that allows real-time imaging of regional ventilation. The purpose of this study was to 1) identify characteristic changes in the EIT signals associated with pneumothoraces; 2) develop and fine-tune an algorithm for their automatic detection; and 3) prospectively evaluate this algorithm for its sensitivity and specificity in detecting pneumothoraces in real time. Design: Prospective controlled laboratory animal investigation. Setting: Experimental Pulmonology Laboratory of the University of Sao Paulo. Subjects: Thirty-nine anesthetized mechanically ventilated supine pigs (31.0 +/- 3.2 kg, mean +/- SD). Interventions. In a first group of 18 animals monitored by EIT, we either injected progressive amounts of air (from 20 to 500 mL) through chest tubes or applied large positive end-expiratory pressure (PEEP) increments to simulate extreme lung overdistension. This first data set was used to calibrate an EIT-based pneumothorax detection algorithm. Subsequently, we evaluated the real-time performance of the detection algorithm in 21 additional animals (with normal or preinjured lungs), submitted to multiple ventilatory interventions or traumatic punctures of the lung. Measurements and Main Results: Primary EIT relative images were acquired online (50 images/sec) and processed according to a few imaging-analysis routines running automatically and in parallel. Pneumothoraces as small as 20 mL could be detected with a sensitivity of 100% and specificity 95% and could be easily distinguished from parenchymal overdistension induced by PEEP or recruiting maneuvers, Their location was correctly identified in all cases, with a total delay of only three respiratory cycles. Conclusions. We created an EIT-based algorithm capable of detecting early signs of pneumothoraces in high-risk situations, which also identifies its location. It requires that the pneumothorax occurs or enlarges at least minimally during the monitoring period. Such detection was operator-free and in quasi real-time, opening opportunities for improving patient safety during mechanical ventilation.