57 resultados para Real-time qPCR


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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 (r 2=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.

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The detection of lane boundaries on suburban streets using images obtained from video constitutes a challenging task. This is mainly due to the difficulties associated with estimating the complex geometric structure of lane boundaries, the quality of lane markings as a result of wear, occlusions by traffic, and shadows caused by road-side trees and structures. Most of the existing techniques for lane boundary detection employ a single visual cue and will only work under certain conditions and where there are clear lane markings. Also, better results are achieved when there are no other onroad objects present. This paper extends our previous work and discusses a novel lane boundary detection algorithm specifically addressing the abovementioned issues through the integration of two visual cues. The first visual cue is based on stripe-like features found on lane lines extracted using a two-dimensional symmetric Gabor filter. The second visual cue is based on a texture characteristic determined using the entropy measure of the predefined neighbourhood around a lane boundary line. The visual cues are then integrated using a rulebased classifier which incorporates a modified sequential covering algorithm to improve robustness. To separate lane boundary lines from other similar features, a road mask is generated using road chromaticity values estimated from CIE L*a*b* colour transformation. Extraneous points around lane boundary lines are then removed by an outlier removal procedure based on studentized residuals. The lane boundary lines are then modelled with Bezier spline curves. To validate the algorithm, extensive experimental evaluation was carried out on suburban streets and the results are presented. 

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Background: Although the pressure flow theory is widely accepted for the transport of photoassimilates in phloem sieve elements, it still requires strong experimental validation. One reason for that is the lack of a precise method for measuring the real-time phloem turgor pressure from the sink tissues, especially in tree trunks. Results: Taking the merits of Hevea brasiliensis, a novel phloem turgor pressure probe based on the state of the art cell pressure probe was developed. Our field measurements showed that the phloem turgor pressure probe can sensitively measure the real-time variation of phloem turgor pressure in H. brasiliensis but the calculation of phloem turgor pressure with xylem tension, xylem sap osmotic potential and phloem sap osmotic potential will under-estimate it. The measured phloem turgor pressure gradient in H. brasiliensis is contrary to the Münch theory. The phloem turgor pressure of H. brasiliensis varied from 8-12 bar as a consequence of water withdrawal from transpiration. Tapping could result in a sharp decrease of phloem turgor pressure followed by a recovery from 8-45 min after the tapping. The recovery of phloem turgor pressure after tapping and its change with xylem sap flow suggest the importance of phloem water relationship in the phloem turgor pressure regulation. Conclusion: The phloem turgor pressure probe is a reliable technique for measuring the real-time variation of phloem turgor pressures in H. brasiliensis. The technique could probably be extended to the accurate measurement of phloem turgor pressure in other woody plants which is essential to test the Münch theory and to investigate the phloem water relationship and turgor pressure regulation. © 2014 An et al.

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On-line education in engineering has attracted a great deal of interest in recent years. One of the difficulties faced in an on-line engineering program is how to ensure effective communication between lecturers and students, and among the students themselves.Techniques common ten years ago such as email, lecture notes posted to websites, and telephone conversations, are now seen as archaic when compared with opportunities offered by more modern communication technologies, such as real-time web-conferencing.We present our efforts to use the web-conferencing software Elluminate-Live! for delivering tutorials, discussion classes, and even laboratory practicals to groups of students studying engineering off-campus, including students posted overseas. Examples are given from two disciplines. We then compare student feedback across all engineering subjects over the years 2012-2013. Our results show that students welcome web-conferencing as a very effectivemeans to deliver classes to distance students and improve their learning experience.

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Coughing and Clapping: Investigating Audience Experience explores the processes and experiences of attending live music events from the initial decision to attend through to audience responses and memories of a performance after it has happened. The book brings together international researchers who consider the experience of being an audience member from a range of theoretical and empirical perspectives. Whether enjoying a drink at a jazz gig, tweeting at a pop concert or suppressing a cough at a classical recital, audience experience is affected by motivation, performance quality, social atmosphere and group and personal identity. Drawing on the implications of these experiences and attitudes, the authors consider the question of what makes an audience, and argue convincingly for the practical and academic value of that question.

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In this work we examine the reliability and validity (in comparison to magnetic resonance imaging; MRI) of real-time ultrasound measures of lumbar erector spinae thickness. We also consider the between-day reliability of the lumbar multifidus muscle area as measured via ultrasound. 23 male subjects aged 21-45 years were measured three times over the course of nine days by one operator. The first (L1) through to the fifth (L5) lumbar vertebral levels were measured on the left and right sides. MRI was performed on the same day as first ultrasound scanning. For between-day intra-rater reliability, intra-class correlation co-efficients (ICCs), standard error of the measurement, minimal detectable difference and co-efficients of variation (CVs) were calculated along with their 95% confidence intervals and Bland-Altman analysis was performed. On Bland-Altman analysis, erector spinae thickness and multifidus area ultrasound measures 'agreed' with equivalent MR measures, though the correlation between MR and ultrasound measures was typically poor to moderate. For both ultrasound measures, the ICCs ranged from 'moderate' to 'excellent' at individual vertebral levels, although multifidus area (CV ranged from 8 to 15%) was less reliable than erector spinae thickness (CV ranged from 6 to 10%). 'Agreement' on Bland-Altmann analysis was present between days for all ultrasound measures. Averaging between sides and between vertebral levels improved reliability. Average erector spinae thickness showed a CV of 5.5% (ICC 0.77) and average multifidus area 6.2% (ICC 0.80).

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We describe an alternative electrochemical technique to monitor covalent bond formation in real-time using nanoparticle-electrode collisions. The method is based on recognising the redox current when MP-11 functionalised chemical reduced graphene oxide (rGO) nanosheets collide with Lomant's reagent modified gold microelectrode. This facile and highly sensitive monitoring method can be useful for investigating the fundamental of single-molecule reactions.

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The success of cloud computing makes an increasing number of real-time applications such as signal processing and weather forecasting run in the cloud. Meanwhile, scheduling for real-time tasks is playing an essential role for a cloud provider to maintain its quality of service and enhance the system's performance. In this paper, we devise a novel agent-based scheduling mechanism in cloud computing environment to allocate real-time tasks and dynamically provision resources. In contrast to traditional contract net protocols, we employ a bidirectional announcement-bidding mechanism and the collaborative process consists of three phases, i.e., basic matching phase, forward announcement-bidding phase and backward announcement-bidding phase. Moreover, the elasticity is sufficiently considered while scheduling by dynamically adding virtual machines to improve schedulability. Furthermore, we design calculation rules of the bidding values in both forward and backward announcement-bidding phases and two heuristics for selecting contractors. On the basis of the bidirectional announcement-bidding mechanism, we propose an agent-based dynamic scheduling algorithm named ANGEL for real-time, independent and aperiodic tasks in clouds. Extensive experiments are conducted on CloudSim platform by injecting random synthetic workloads and the workloads from the last version of the Google cloud tracelogs to evaluate the performance of our ANGEL. The experimental results indicate that ANGEL can efficiently solve the real-time task scheduling problem in virtualized clouds.

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As clouds have been deployed widely in various fields, the reliability and availability of clouds become the major concern of cloud service providers and users. Thereby, fault tolerance in clouds receives a great deal of attention in both industry and academia, especially for real-time applications due to their safety critical nature. Large amounts of researches have been conducted to realize fault tolerance in distributed systems, among which fault-tolerant scheduling plays a significant role. However, few researches on the fault-tolerant scheduling study the virtualization and the elasticity, two key features of clouds, sufficiently. To address this issue, this paper presents a fault-tolerant mechanism which extends the primary-backup model to incorporate the features of clouds. Meanwhile, for the first time, we propose an elastic resource provisioning mechanism in the fault-tolerant context to improve the resource utilization. On the basis of the fault-tolerant mechanism and the elastic resource provisioning mechanism, we design novel fault-tolerant elastic scheduling algorithms for real-time tasks in clouds named FESTAL, aiming at achieving both fault tolerance and high resource utilization in clouds. Extensive experiments injecting with random synthetic workloads as well as the workload from the latest version of the Google cloud tracelogs are conducted by CloudSim to compare FESTAL with three baseline algorithms, i.e., Non-M igration-FESTAL (NMFESTAL), Non-Overlapping-FESTAL (NOFESTAL), and Elastic First Fit (EFF). The experimental results demonstrate that FESTAL is able to effectively enhance the performance of virtualized clouds.