Expression of Aspergillus nidulans phy Gene in Nicotiana benthamiana Produces Active Phytase with Broad Specificities

A full-length phytase gene (phy) of Aspergillus nidulans was amplified from the cDNA library by polymerase chain reaction (PCR), and it was introduced into a bacterial expression vector, pET-28a. The recombinant protein (rPhy-E, 56 kDa) was overexpressed in the insoluble fraction of Escherichia coli culture, purified by Ni-NTA resin under denaturing conditions and injected into rats as an immunogen. To express A. nidulans phytase in a plant, the full-length of phy was cloned into a plant expression binary vector, pPZP212. The resultant construct was tested for its transient expression by Agrobacterium-infiltration into Nicotiana benthamiana leaves. Compared with a control, the agro-infiltrated leaf tissues showed the presence of phy mRNA and its high expression level in N. benthamiana. The recombinant phytase (rPhy-P, 62 kDa) was strongly reacted with the polyclonal antibody against the nonglycosylated rPhy-E. The rPhy-P showed glycosylation, two pH optima (pH 4.5 and pH 5.5), an optimum temperature at 45~55 °C, thermostability and broad substrate specificities. After deglycosylation by peptide-N-glycosidase F (PNGase-F), the rPhy-P significantly lost the phytase activity and retained 1/9 of the original activity after 10 min of incubation at 45 °C. Therefore, the deglycosylation caused a significant reduction in enzyme thermostability. In animal experiments, oral administration of the rPhy-P at 1500 U/kg body weight/day for seven days caused a significant reduction of phosphorus excretion by 16% in rat feces. Besides, the rPhy-P did not result in any toxicological changes and clinical signs.     

Fluorescent Probe DCVJ Shows High Sensitivity for Characterization of Amyloid β-Peptide Early in the Lag Phase

The aggregation of intrinsically disordered and misfolded proteins in the form of oligomers and fibrils plays a crucial role in a number of neurological and neurodegenerative diseases. Currently, most probes and biophysical techniques that detect and characterize fibrils at high resolution fail to show sensitivity and binding for oligomers. Here, we show that 9-(dicyano-vinyl)julolidine (DCVJ), a class of molecular rotor, binds amyloid beta (Aβ) early aggregates, and we report the kinetics as well as packing of the oligomer formation. The binding of DCVJ to Aβ40 increased its emission intensity with time at 510 nm and produced a second excimer peak at 575 nm. However, DCVJ did not bind to the prefibrillar aggregates of Aβ42, which indicated that the oligomers formed by Aβ40 and Aβ42 were not the same. The F4C F19W mutant of Aβ40, which did not form fibrils, also bound DCVJ, but the emission spectral profile varied from that of the wild-type (WT). Atomic force microscopy images of WT Aβ40, the F4C F19W mutant, and Aβ42 oligomers displayed differences in size and shape, confirming the difference in their DCVJ spectra. The effect of epigallocatechin-3-gallate (EGCG) on the reduction of Aβ42 fibrils was also observed with finer detail than with other techniques. The results of this study show that DCVJ detects early aggregates and provides valuable information regarding the oligomer kinetics, packing, and mechanism of formation.     

A review and new approach to minimize the cost of solar assisted absorption cooling system

Solar energy is an alternative energy source for cooling systems where electricity is demand or expensive. Many solar assisted cooling systems have been installed in different countries for domestic purpose. Many researches are going on to achieve economical and efficient thermal systems when compared with conventional systems. This paper reviews the past efforts of solar assisted-single effect vapour absorption cooling system using LiBr–H₂O mixture for residential buildings. Solar assisted single-effect absorption cooling systems were capable of working in the driving temperature range of 70–100 °C. In this system LiBr–H₂O are the major working pairs and has a higher COP than any other working fluids. Besides the review of the past theoretical and experimental investigations of solar single effect absorption cooling systems, some new ideas were introduced to minimize the capital and operational cost, to reduce heat loss from generator and thus to increase COP to get effective cooling.     

Every team deserves a second chance: an extended study on predicting team performance

Voting among different agents is a powerful tool in problem solving, and it has been widely applied to improve the performance in finding the correct answer to complex problems. We present a novel benefit of voting, that has not been observed before: we can use the voting patterns to assess the performance of a team and predict their final outcome. This prediction can be executed at any moment during problem-solving and it is completely domain independent. Hence, it can be used to identify when a team is failing, allowing an operator to take remedial procedures (such as changing team members, the voting rule, or increasing the allocation of resources). We present three main theoretical results: (1) we show a theoretical explanation of why our prediction method works; (2) contrary to what would be expected based on a simpler explanation using classical voting models, we show that we can make accurate predictions irrespective of the strength (i.e., performance) of the teams, and that in fact, the prediction can work better for diverse teams composed of different agents than uniform teams made of copies of the best agent; (3) we show that the quality of our prediction increases with the size of the action space. We perform extensive experimentation in two different domains: Computer Go and Ensemble Learning. In Computer Go, we obtain high quality predictions about the final outcome of games. We analyze the prediction accuracy for three different teams with different levels of diversity and strength, and show that the prediction works significantly better for a diverse team. Additionally, we show that our method still works well when trained with games against one adversary, but tested with games against another, showing the generality of the learned functions. Moreover, we evaluate four different board sizes, and experimentally confirm better predictions in larger board sizes. We analyze in detail the learned prediction functions, and how they change according to each team and action space size. In order to show that our method is domain independent, we also present results in Ensemble Learning, where we make online predictions about the performance of a team of classifiers, while they are voting to classify sets of items. We study a set of classical classification algorithms from machine learning, in a data-set of hand-written digits, and we are able to make high-quality predictions about the final performance of two different teams. Since our approach is domain independent, it can be easily applied to a variety of other domains.     

Modulation of Cellular Fate of Vinyl Triarylamines through Structural Fine Tuning: To Stay or Not To Stay in the Mitochondria?

Mitochondria are involved in many cellular pathways and dysfunctional mitochondria are linked to various diseases. Hence efforts have been made to design mitochondria-targeted fluorophores for monitoring the mitochondrial status. However, the factors that govern the mitochondria-targeted potential of dyes are not well-understood. In this context, we synthesized analogues of the TP-2Bzim probe belonging to the vinyltriphenylamine (TPA) class and already described for its capacity to bind nuclear DNA in fixed cells and mitochondria in live cells. These analogues ( TP-1Bzim, TPⁿ-2Bzim, TP¹⁺-2Bzim, TN-2Bzim ) differ in the cationic charge, the number of vinylbenzimidazolium branches and the nature of the triaryl core. Using microscopy, we demonstrated that the cationic derivatives accumulate in mitochondria but do not reach mtDNA. Under depolarisation of the mitochondrial membrane, TP-2Bzim and TP¹⁺-2Bzim translocate to the nucleus in direct correlation with their strong DNA affinity. This reversible phenomenon emphasizes that these probes can be used to monitor ΔΨ_m variations.     

Technical note Temporal remote sensing data and GIS application in landslide hazard zonation of part of Western ghat,India

Spatial and temporal multi-layered information is required to assess landslide hazard susceptibility. The manual method of data integration for targeting potential zones susceptible to landslide hazard is time consuming. The present study highlights the utility of temporal remotely-sensed data and knowledgebased Geographical Information Systems for collection, integration and analysis of spatially-oriented data, as well as in finding out the inherent relation between separate entities in parts of West ghat in India.     

Interaction Between Two Mutually Orthogonal Heat-Generating Baffles in a Square Cavity

Laminar free convection induced by two mutually orthogonal discrete heat-generating baffles in a two-dimensional square cavity is analyzed numerically. The computations were carried out for different locations and combinations of heat source strengths of the baffles for a fixed Grashof number of 10⁶and Prandtl number of 0.71. The coupled governing equations were solved bya finite-difference method using alternating direction implicit technique and successive overrelaxation methods. The obtained results clearly show that the hydrodynamic and thermal fields in the cavity depend on both the location and strength of the heat-generating baffles. Though the flow inhibition is caused by both the baffles, the baffle with higher source strength plays a decisive role in inducing the flow. The locations of baffles with unequal source strengths produce significant changes in the net heat transfer rate. This is further magnified for higher contrast in source strengths. This study provides qualitative suggestions that may improve the thermal design of sealed enclosures, which are encountered frequently in the electronics industry.     

Error diffusion in cell-oriented Fourier-transform computer-generated holograms to compensate for printing constraints

Abstract

Error-diffused quantization has been applied to the generation of cell-oriented computer-generated Fourier transform holograms, resulting in reduced reconstruction errors. Improvements are demonstrated when applied to the algorithms of Lohmann, Lee and Burckhardt after invoking realistic constraints on the minimum size of the printable spot for an electrophotographic laser printer.