Prevalence of beak and feather disease virus in wild Platycercus elegans: comparison of three tissue types using a probe-based real-time qPCR test

The detection of avian viruses in wild populations has considerable conservation implications. For DNA-based studies, feathers may be a convenient sample type for virus screening and are, therefore, an increasingly common technique. This is despite recent concerns about DNA quality, ethics, and a paucity of data comparing the reliability and sensitivity of feather sampling to other common sample types such as blood. Alternatively, skeletal muscle tissue may offer a convenient sample to collect from dead birds, which may reveal viraemia. Here, we describe a probe-based quantitative real-time PCR for the relative quantification of beak and feather disease virus (BFDV), a pathogen of serious conservation concern for parrots globally. We used this method to test for BFDV in wild crimson rosellas (Platycercus elegans), and compared three different sample types. We detected BFDV in samples from 29 out of 84 individuals (34.5%). However, feather samples provided discordant results concerning virus presence when compared with muscle tissue and blood, and estimates of viral load varied somewhat between different sample types. This study provides evidence for widespread infection of BFDV in wild crimson rosellas, but highlights the importance of sample type when generating and interpreting qualitative and quantitative avian virus data.

Effect of creatine supplementation on housekeeping genes in human skeletal muscle using real-time RT-PCR

The present study examined the validity and reliability of measuring the expression of various genes in human skeletal muscle using quantitative real-time RT-PCR on a GeneAmp 5700 sequence detection system with SYBR Green 1 chemistry. In addition, the validity of using some of these genes as endogenous controls (i.e., housekeeping genes) when human skeletal muscle was exposed to elevated total creatine levels and exercise was also examined. For all except 28S, linear relationships between the logarithm of the starting RNA concentrations and the cycle threshold (C_T) values were established for ß-actin, ß2-microglobulin (ß₂M), cyclophilin (CYC), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). We found a linear response between C_T values and the logarithm of a given amount of starting cDNA for all the genes tested. The overall intra-assay coefficient of variance for these genes was 1.3% and 21% for raw C_T values and the linear value of 2^-C_T, respectively. Interassay variability was 2.3% for raw C_T values and 34% for the linear value of 2^-C_T. We also examined the expression of various housekeeping genes in human skeletal muscle at days 0, 1, and 5 following oral supplementation with either creatine or a placebo employing a double-blind crossover study design. Treatments were separated by a 5-wk washout period. Immediately following each muscle sampling, subjects performed two 30-s all-out bouts on a cycle ergometer. Creatine supplementation increased (P < 0.05) muscle total creatine content above placebo levels; however, there were no changes (P > 0.05) in C_T values across the supplementation periods for any of the genes. Nevertheless, 95% confidence intervals showed that GAPDH was variable, whereas ß-actin, ß₂M, and CYC were the least varying genes. Normalization of the data to these housekeeping genes revealed variable behavior for ß₂M with more stable expressions for both ß-actin and CYC. We conclude that, using real-time RT-PCR, ß-actin or CYC may be used as housekeeping genes to study gene expression in human muscle in experiments employing short-term creatine supplementation combined with high-intensity exercise.

Providing real-time information services for public bus commuters

This paper investigates RTIS for public bus commuters; identifying their needs and proposing the architecture and design for this service. Considering that the service needs to be extensible, adaptable and interoperative, the new system is based on the use of XML Web Services. This gives the structure flexibility for further development and integration with other like services.

Miscibility, crystallization κinetics and real-time small-Angle x-ray scattering investigation of the semicrystalline morphology in thermosetting polymer blends of epoxy resin and poly(ethylene oxide)

Thermosetting polymer blends of poly(ethylene oxide) (PEO) and bisphenol-A-type epoxy resin (ER) were prepared using 4,4′-methylenebis(3-chloro-2,6-diethylaniline) (MCDEA) as curing agent. The miscibility and crystallization behavior of MCDEA-cured ER/PEO blends were investigated by differential scanning calorimetry (DSC). The existence of a single composition-dependent glass transition temperature (Tg) indicates that PEO is completely miscible with MCDEA-cured ER in the melt and in the amorphous state over the entire composition range. Fourier-transform infrared (FTIR) investigations indicated hydrogen-bonding interaction between the hydroxyl groups of MCDEA-cured ER and the ether oxygens of PEO in the blends, which is an important driving force for the miscibility of the blends. The average strength of the hydrogen bond in the cured ER/PEO blends is higher than in the pure MCDEA-cured ER. Crystallization kinetics of PEO from the melt is strongly influenced by the blend composition and the crystallization temperature. At high conversion, the time dependence of the relative degree of crystallinity deviated from the Avrami equation. The addition of a non-crystallizable ER component into PEO causes a depression of both the overall crystallization rate and the melting temperature. The surface free energy of folding σe displays a minimum with variation of composition. The spherulitic morphology of PEO in the ER/PEO blends exhibits typical characteristics of miscible crystalline/amorphous blends, and the PEO spherulites in the blends are always completely volume-filling. Real-time small-angle X-ray scattering (SAXS) experiments reveal that the long period L increases drastically with increasing ER content at the same temperatures. The amorphous cured ER component segregates interlamellarly during the crystallization process of PEO because of the low chain mobility of the cured ER. A model describing the semicrystalline morphology of MCDEA-cured ER/PEO blends is proposed based on the SAXS results. The semicrystalline morphology is a stack of crystalline lamellae; the amorphous fraction of PEO, the branched ER chains and imperfect ER network are located between PEO lamellae.

Real time, perceived time and time online : a review of student experiences of time over thirty years of distance and online education

This paper examines how students use and perceive time when studying in distance education modes and what affects this perception of time and the reality of time. We examine 30 years of student involvement on distance and online education, their comments on both their learning experiences, and the technology requirements of distance education/online learning. Our University has been involved in distance education since its formation in 1974. The online technologies offer increasingly sophisticated and immersive experiences for our students, both on campus and off campus, but many of our students continue to complain of time squeeze, and fail to predict the time it will take them to complete our subjects. We research how the technologies we use for online learning are contributing to this time squeeze perception and the student's "real" time to learn.
Research is drawn from both the Australian Bureau of Statistics and surveys of our students' experiences (we have 32,000 students online, with single online classes of over 1300 students), to examine student use and perceptions of their available time to study and how the technologies used in online learning affect this.

Poesis in diegesis : real-time interactive dance performance

This paper describes the processes of creating a hybrid experience of live performance and virtual imagery in 1 + x: Mid-range Projections, a dance work that uses real-time processing of live feed video to create a duet between dancers and their virtual images. 1 + x is about creating experiences of interiority and intimacy, ironically, through the juxtaposition of virtual images of performers with their real selves. This paper examines how the ideal of reinserting intimacy, humanity and 'presence' into a technologically generated image space, in the case of 1 + x, depends on the precise and delicate manipulation of live-feed video in collaboration with the audience's experience of interactivity. The result is a performance work that creates a poesis of 'presence', through a diegesis that melds real and virtual images on the same screen and the same conceptual and spatial plane - a 'becoming virtual'.

The brachyspira hyodysenteriae ftnA gene : DNA vaccination and real-time PCR quantification of bacteria in a mouse model of disease

The nucleotide sequence of the Brachyspira hyodysenteriae ftnA gene, encoding a putative ferritin protein (FtnA), was determined. Analysis of the sequence predicted that this gene encoded a protein of 180 amino acids. RT-PCR and Western blot showed that the ftnA gene was expressed in B. hyodysenteriae, and evidence suggests that FtnA stores iron rather than haem. ftnA was delivered as DNA and recombinant protein vaccines in a mouse model of B. hyodysenteriae infection. Vaccine efficacy was monitored by caecal pathology and quantification of B. hyodysenteriae numbers in the caeca of infected mice by real-time PCR.

A video-based real-time vehicle detection method by classified background learning

A new two-level real-time vehicle detection method is proposed in order to meet the robustness and efficiency requirements of real world applications. At the high level, pixels of the background image are classified into three categories according to the characteristics of Red, Green, Blue (RGB) curves. The robustness of the classification is further enhanced by using
line detection and pattern connectivity. At the lower level, an exponential forgetting algorithm with adaptive parameters for different categories is utilised to calculate the background and reduce the distortion by the small motion of video cameras. Scene tests show that the proposed method is more robust and faster than previous methods, which is very suitable for real-time vehicle detection in outdoor environments, especially concerning locations where the level of illumination changes frequently and speed detection is important.

RFID-based real-time smart waste management system

In an environmental context, the use of RFID (radio frequency identification) and load cell sensor technology can be employed for not only bringing down waste management costs, but also to facilitate automating and streamlining waste (e.g., garbage, recycling, and green) identification and weight measurement processes for designing smart waste management systems. In this paper, we outline a RFID and sensor model for designing a system in real-time waste management. An application of the architecture is described in the area of RFID and sensor based automatic waste identity, weight, and stolen bins identification system (WIWSBIS).

Delay control and parallel admission algorithms for real-time anycast flow

An anycast flow is a flow that can be connected to any one of the members in a group of designated (replicated) servers (called anycast group). In this paper, we derive a set of formulas for calculating the end-to-end delay bound for the anycast flows and present novel admission control algorithms for anycast flows with real-time constraints. Given such an anycast group, our algorithms can effectively select the paths for anycast flows' admission and connection based on the least end-to-end delay bounds evaluated. We also present a parallel admission control algorithm that can effectively calculate the available paths with a short delay bound for different destinations in the anycast group so that a best path with the shortest delay bound can be chosen.

Real time RT-PCR analysis of housekeeping genes in human skeletal muscle following acute exercise

Studies examining gene expression with RT-PCR typically normalize their mRNA data to a constitutively expressed housekeeping gene. The validity of a particular housekeeping gene must be determined for each experimental intervention. We examined the expression of various housekeeping genes following an acute bout of endurance (END) or resistance (RES) exercise. Twenty-four healthy subjects performed either a interval-type cycle ergometry workout to exhaustion (~75 min; END) or 300 single-leg eccentric contractions (RES). Muscle biopsies were taken before exercise and 3 h and 48 h following exercise. Real-time RT-PCR was performed on ß-actin, cyclophilin (CYC), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and ß2-microglobulin (ß2M). In a second study, 10 healthy subjects performed 90 min of cycle ergometry at ~65% of O2 max, and we examined a fifth housekeeping gene, 28S rRNA, and reexamined ß2M, from muscle biopsy samples taken immediately postexercise. We showed that CYC increased 48 h following both END and RES exercise (3- and 5-fold, respectively; P < 0.01), and 28S rRNA increased immediately following END exercise (2-fold; P = 0.02). ß-Actin trended toward an increase following END exercise (1.85-fold collapsed across time; P = 0.13), and GAPDH trended toward a small yet robust increase at 3 h following RES exercise (1.4-fold; P = 0.067). In contrast, ß2M was not altered at any time point postexercise. We conclude that ß2M and ß-actin are the most stably expressed housekeeping genes in skeletal muscle following RES exercise, whereas ß2M and GAPDH are the most stably expressed following END exercise.

Performance optimization for energy-aware adaptive checkpointing in embedded real-time systems

Using additional store-checkpoinsts (SCPs) and compare-checkpoints (CCPs), we present an adaptive checkpointing for double modular redundancy (DMR) in this paper. The proposed approach can dynamically adjust the checkpoint intervals. We also design methods to calculate the optimal numbers of checkpoints, which can minimize the average execution time of tasks. Further, the adaptive checkpointing is combined with the DVS (dynamic voltage scaling) scheme to achieve energy reduction. Simulation results show that, compared with the previous methods, the proposed approach significantly increases the likelihood of timely task completion and reduces energy consumption in the presence of faults.