Detection of avian influenza virus by fluorescent DNA barcode-based immunoassay with sensitivity comparable to PCR

In this paper, a coupling of fluorophore-DNA barcode and bead-based immunoassay for detecting avian influenza virus (AIV) with PCR-like sensitivity is reported. The assay is based on the use of sandwich immunoassay and fluorophore-tagged oligonucleotides as representative barcodes. The detection involves the sandwiching of the target AIV between magnetic immunoprobes and barcode-carrying immunoprobes. Because each barcode-carrying immunoprobe is functionalized with a multitude of fluorophore-DNA barcode strands, many DNA barcodes are released for each positive binding event resulting in amplification of the signal. Using an inactivated H16N3 AIV as a model, a linear response over five orders of magnitude was obtained, and the sensitivity of the detection was comparable to conventional RT-PCR. Moreover, the entire detection required less than 2 hr. The results indicate that the method has great potential as an alternative for surveillance of epidemic outbreaks caused by AIV, other viruses and microorganisms.

Detection of Avian Influenza Virus with PCR-Like Sensitivity by Fluorescent DNA Barcode-Based Immunoassay

In this paper, we report a coupling of fluorophore-DNA barcode and bead-based
immunoassay for the detection of Avian Influenza Virus (AIV), a potential pandemic threat for human health and enormous economic losses. The detection strategy is based on the use of sandwich immunoassay and fluorophore-tagged oligonucleotides as representatively fluorescent barcodes. Despite its simplicity the assay has sensitivity comparable to RT-PCR amplification, and possesses a great potential as a rapid and sensitive on-chip detection format.

Rhinovirus upregulates transient receptor potential channels in a human neuronal cell line: Implications for respiratory virus-induced cough reflex sensitivity

ABSTRACT (250 words)
BACKGROUND: The mechanism underlying respiratory virus-induced cough hypersensitivity is unknown. Up-regulation of airway neuronal receptors responsible for sensing physical and chemical stimuli is one possibility and the transient receptor potential (TRP) channel family are potential candidates. We have used an in vitro model of sensory neurones and human rhinovirus (HRV-16) to study the effect of virus infection on TRP expression.
METHODS: IMR32 neuroblastoma cells were differentiated in culture to express three TRP channels, TRPV1, TRPA1 and TRPM8. Flow cytometry and qRT-PCR were used to measure TRP channel protein and mRNA levels following inoculation with live virus, inactivated virus, virus- induced soluble factors or pelleted virus particles. Multiplex bioassay was used to determine nerve growth factor (NGF), interleukin (IL)-1ß, IL-6 and IL-8 levels in response to infection.
RESULTS: Early up-regulation of TRPA1 and TRPV1 expression occurred 2 to4 hours post infection. This was independent of replicating virus as virus induced soluble factors alone were sufficient to increase channel expression 50 and 15 fold, respectively. NGF, IL-6 and IL-8 levels, increased in infected cell supernatants, represent possible candidates. In contrast, TRPM8 expression was maximal at 48 hours (9.6 fold) and required virus replication rather than soluble factors
CONCLUSIONS We show for the first time that rhinovirus can infect neuronal cells. Furthermore, infection causes up-regulation of TRP channels by channel specific mechanisms. Increase in TRPA1 and TRPV1 levels can be mediated by soluble factors induced by infection whereas TRPM8 requires replicating virus. TRP channels may be novel therapeutic targets for controlling virus-induced cough.

Analysis of Load Impact on Small-signal Stability based on Load Active Power Sensitivity

While load flow conditions vary with different loads, the small-signal stability of the entire system is closely related with to the locations, capacities and models of loads. In this paper, load impacts with different capacities and models on the small-signal stability are analysed. In the real large-scale power system case, the load sensitivity which denotes the sensitivity of the eigenvalue with respect to the load active power is introduced and applied to rank the loads. The loads with high sensitivity are also considered.

High population prevalence of cardiac troponin I measured by a high-sensitivity assay and cardiovascular risk estimation: the MORGAM Biomarker Project Scottish Cohort

Aims
Our aim was to test the prediction and clinical applicability of high-sensitivity assayed troponin I for incident cardiovascular events in a general middle-aged European population.

Methods and results
High-sensitivity assayed troponin I was measured in the Scottish Heart Health Extended Cohort (n = 15 340) with 2171 cardiovascular events (including acute coronary heart disease and probable ischaemic strokes), 714 coronary deaths (25% of all deaths), 1980 myocardial infarctions, and 797 strokes of all kinds during an average of 20 years follow-up. Detection rate above the limit of detection (LoD) was 74.8% in the overall population and 82.6% in men and 67.0% in women. Troponin I assayed by the high-sensitivity method was associated with future cardiovascular risk after full adjustment such as that individuals in the fourth category had 2.5 times the risk compared with those without detectable troponin I (P < 0.0001). These associations remained significant even for those individuals in whom levels of contemporary-sensitivity troponin I measures were not detectable. Addition of troponin I levels to clinical variables led to significant increases in risk prediction with significant improvement of the c-statistic (P < 0.0001) and net reclassification (P < 0.0001). A threshold of 4.7 pg/mL in women and 7.0 pg/mL in men is suggested to detect individuals at high risk for future cardiovascular events.

Conclusion
Troponin I, measured with a high-sensitivity assay, is an independent predictor of cardiovascular events and might support selection of at risk individuals.

Investigation of moisture condition and Autoclam sensitivity on air permeability measurements for both normal concrete and high performance concrete

While on site measurement of air permeability provides a useful approach for assessing the likely long term durability of concrete structures, no existing test method is capable of effectively determining the relative permeability of high performance concrete (HPC). Lack of instrument sensitivity and the influence of concrete moisture are proposed as two key reasons for this phenomenon. With limited systematic research carried out in this area to date, the aim if this study was to investigate the influence of instrument sensitivity and moisture condition on air permeability measurements for both normal concrete and HPC. To achieve a range of moisture conditions, samples were dried initially for between one and 5 weeks and then sealed in polythene sheeting and stored in an oven at 50 C to internally distribute moisture evenly. Moisture distribution was determined throughout using relative humidity probe and electrical resistance measurements. Concrete air permeability was subsequently measured using standardised air permeability (Autoclam) and water penetration (BS EN: 12390-8) tests to assess differences between the HPCs tested in this study. It was found that for both normal and high performance concrete, the influence of moisture on Autoclam air permeability results could be eliminated by pre-drying (50 ± 1 C, RH 35%) specimens for 3 weeks. While drying for 5 weeks alone was found not to result in uniform internal moisture distributions, this state was achieved by exposing specimens to a further 3 weeks of sealed pre-conditioning at 50 ± 1 C. While the Autoclam test was not able to accurately identify relative HPC quality due to low sensitivity at associated performance levels, an effective preconditioning procedure to obtain reliable air permeability of HPC concretes was identified. © 2013 The Authors

Multibody modelling of a TB31 and a TB32 crash test with vertical portable concrete barriers: model verification and sensitivity analysis.

In this article the multibody simulation software package MADYMO for analysing and optimizing occupant safety design was used to model crash tests for Normal Containment barriers in accordance with EN 1317. The verification process was carried out by simulating a TB31 and a TB32 crash test performed on vertical portable concrete barriers and by comparing the numerical results to those obtained experimentally. The same modelling approach was applied to both tests to evaluate the predictive capacity of the modelling at two different impact speeds. A sensitivity analysis of the vehicle stiffness was also carried out. The capacity to predict all of the principal EN1317 criteria was assessed for the first time: the acceleration severity index, the theoretical head impact velocity, the barrier working width and the vehicle exit box. Results showed a maximum error of 6% for the acceleration severity index and 21% for theoretical head impact velocity for the numerical simulation in comparison to the recorded data. The exit box position was predicted with a maximum error of 4°. For the working width, a large percentage difference was observed for test TB31 due to the small absolute value of the barrier deflection but the results were well within the limit value from the standard for both tests. The sensitivity analysis showed the robustness of the modelling with respect to contact stiffness increase of ±20% and ±40%. This is the first multibody model of portable concrete barriers that can reproduce not only the acceleration severity index but all the test criteria of EN 1317 and is therefore a valuable tool for new product development and for injury biomechanics research.

A Sensitivity Approach for Eliminating Clashes from Computer Aided Design Model Assemblies

Clashes occur when components in an assembly unintentionally violate others. If clashes are not identified and designed out before manufacture, product function will be reduced or substantial cost will be incurred in rework. This paper introduces a novel approach for eliminating clashes by identifying which parameters defining the part features in a computer aided design (CAD) assembly need to change and by how much. Sensitivities are calculated for each parameter defining the part and the assembly as the change in clash volume due to a change in each parameter value. These sensitivities give an indication of important parameters and are used to predict the optimum combination of changes in each parameter to eliminate the clash. Consideration is given to the fact that it is sometimes preferable to modify some components in an assembly rather than others and that some components in an assembly cannot be modified as the designer does not have control over their shape. Successful elimination of clashes has been demonstrated in a number of example assemblies.

Simple high sensitivity wireless transceiver

This article describes an extremely simple wireless transceiver, comprising of only a low Q VCO and a phase locked loop IC. It is experimentally shown to, simultaneously, transmit an 8-dBm CW interrogation signal, while concurrently demodulating a phase modulated received signal with sensitivity levels of -120 dBm. This makes the performance similar to conventional transceivers, which require complex superheterodyne type architectures and also require a means to provide a high isolation separate the transmit/receive signals (such as a circulator). 

Distribution Network Voltage Support Using Sensitivity-based Dispatch of Distributed Generation

This paper is concerned with the voltage and reactive power issues surrounding the connection of Distributed Generation (DG) on the low-voltage (LV) distribution network. The presented system-wide voltage control algorithm consists of three stages. Firstly available reactive power reserves are utilized. Then, if required, DG active power output is curtailed. Finally, curtailment of non-critical site demand is considered. The control methodology is tested on a variant of the 13-bus IEEE Node Radial Distribution Test Feeder. The presented control algorithm demonstrated that the distribution system operator (DSO) can maintain voltage levels within a desired statutory range by dispatching reactive power from DG or network devices. The practical application of the control strategy is discussed.

Comparison of 5% versus 15% sucrose intakes as part of a eucaloric diet in overweight and obese subjects:effects on insulin sensitivity, glucose metabolism, vascular compliance, body composition and lipid profile. A randomised controlled trial

AIMS: The effect of dietary sucrose on insulin resistance and the pathogenesis of diabetes and vascular disease is unclear. We assessed the effect of 5% versus 15% sucrose intakes as part of a weight maintaining, eucaloric diet in overweight/obese subjects.

METHODS: Thirteen subjects took part in a randomised controlled crossover study (M:F 9:4, median age 46 years, range 37-56 years, BMI 31.7±0.9 kg/m(2)). Subjects completed two 6 week dietary periods separated by 4 week washout. Diets were designed to have identical macronutrient profile. Insulin action was assessed using a two-step hyperinsulinaemic euglycaemic clamp; glucose tolerance, vascular compliance, body composition and lipid profiles were also assessed.

RESULTS: There was no change in weight or body composition between diets. There was no difference in peripheral glucose utilization or suppression of endogenous glucose production. Fasting glucose was significantly lower after the 5% diet. There was no demonstrated effect on lipid profiles, blood pressure or vascular compliance.

CONCLUSION: A low-sucrose diet had no beneficial effect on insulin resistance as measured by the euglycaemic glucose clamp. However, reductions in fasting glucose, one hour insulin and insulin area under the curve with the low sucrose diet on glucose tolerance testing may indicate a beneficial effect and further work is required to determine if this is the case. Clinical Trial Registration number ISRCTN50808730.

Reduction of the temperature sensitivity of Halomonas hydrothermalis by iron starvation combined with microaerobic conditions

The limits to biological processes on Earth are determined by physicochemical parameters, such as extremes of temperature and low water availability. Research into microbial extremophiles has enhanced our understanding of the biophysical boundaries which define the biosphere. However, there remains a paucity of information on the degree to which rates of microbial multiplication within extreme environments are determined by the availability of specific chemical elements. Here, we show that iron availability and composition of the gaseous phase (aerobic vs. microaerobic) determine susceptibility of a marine bacterium, Halomonas hydrothermalis, to sub-optimal and elevated temperature and salinity by impacting rates of cell division (but not viability). In particular, iron starvation combined with microaerobic conditions (5 % v/v of O2, 10 % v/v of CO2, reduced pH) reduced sensitivity to temperature across the 13 °C range tested. These data demonstrate that nutrient limitation interacts with physicochemical parameters to determine biological permissiveness for extreme environments. The interplay between resource availability and stress tolerance, therefore, may shape the distribution and ecology of microorganisms within Earth's biosphere.

Effects of phosphate on arsenate and arsenite sensitivity in two rice (Oryza sativa L.) cultivars of different sensitivity

Arsenate and arsenite sensitivity and arsenate influx tests were conducted for two rice cultivars of different arsenic sensitivity. Azucena and Bala. These were to establish if the mechanism of reduced arsenic sensitivity is achieved through an altered phosphate uptake system, as shown for Holcus lanatus. High phosphate treatments (>= 50 mu M) provided protection against both arsenate and arsenite. Unlike the H. lanatus tolerance mechanism, in the less sensitive cultivar Bala, arsenate influx did not decrease with phosphate treatment and phosphate transporters appeared to be constitutively upregulated; V(max) for arsenate influx remain similar when Bala was grown in the presence or absence of phosphate (V(max) - 0.90 and 0.63 nmol g(-1) f.wt min(-1) respectively). Although mean K(m) appear different, Bala did not show lower affinity to arsenate than Azucena in the absence of phosphate (K(m) - Azucena, 0.30 mM and Bala, 0.18), while in phosphate treatment, Bala arsenate affinity was half that observed for Azucena (K(m) - Azucena, 0.14 and Bala, 0.36 mM). These were low compared to a 4 and 6 fold decrease seen for similar studies on H. lanatus in the absence and presence of phosphate. Phosphate-induced arsenic protection was observed but the mechanism does not resemble that of H. lanatus. Alternative mechanisms were discussed. (C) 2010 Elsevier B.V. All rights reserved.