Engineering of the Escherichia coli Im7 immunity protein as a loop display scaffold

Protein scaffolds derived from non-immunoglobulin sources are increasingly being adapted and engineered to provide unique binding molecules with a diverse range of targeting specificities. The ColE7 immunity protein (Im7) from Escherichia coli is potentially one such molecule, as it combines the advantages of (i) small size, (ii) stability conferred by a conserved four anti-parallel alpha-helical framework and (iii) availability of variable surface loops evolved to inactivate members of the DNase family of bacterial toxins, forming one of the tightest known protein-protein interactions. Here we describe initial cloning and protein expression of Im7 and its cognate partner the 15 kDa DNase domain of the colicin E7. Both proteins were produced efficiently in E.coli, and their in vitro binding interactions were validated using ELISA and biosensor. In order to assess the capacity of the Im7 protein to accommodate extensive loop region modifications, we performed extensive molecular modelling and constructed a series of loop graft variants, based on transfer of the extended CDR3 loop from the IgG1b12 antibody, which targets the gp120 antigen from HIV-1. Loop grafting in various configurations resulted in chimeric proteins exhibiting retention of the underlying framework conformation, as measured using far-UV circular dichroism spectroscopy. Importantly, there was low but measurable transfer of antigen-specific affinity. Finally, to validate Im7 as a selectable scaffold for the generation of molecular libraries, we displayed Im7 as a gene 3 fusion protein on the surface of fd bacteriophages, the most common library display format. The fusion was successfully detected using an anti-Im7 rabbit polyclonal antibody, and the recombinant phage specifically recognized the immobilized DNase. Thus, Im7 scaffold is an ideal protein display scaffold for the future generation and for the selection of libraries of novel binding proteins.     

Electrospray mass spectrometry for characterizing polyglycerols and the effects of adduct ion and cone voltage

Polyglycerol intermediates have been characterized by liquid chromatography-mass spectrometry (LC-MS) with electrospray ionization (ESI). Linear and cyclic components from n=2–23 in a sample of decaglycerol, for example, have been resolved in the second dimension or mass axis. Molecular weight (MW) distributions for tri-, hexa-, and decaglycerol products have been analyzed as a function of cone voltage and adduct ion (H⁺, Li⁺, Na⁺, K⁺, Rb⁺, Cs⁺, and NH₄ ⁺). A different combination is required to obtain a reliable MW distribution for each polyglycerol intermediate. The best distribution obtained by ESI/MS is determined by comparing the calculated hydroxyl number and cyclic content to that obtained by wet chemistry and gas chromatography, respectively. Once ESI/MS conditions are established, the distribution can be used, for the first time in polyglycerol analysis, to calculate important parameters such as number average MW, weight average MW, polydispersity, % cyclics, hydroxyl number, wt% above n=6, etc.     

Polymer-graphene nanocomposites as ultrafast-charge and -discharge cathodes for rechargeable lithium batteries

Electroactive polymers are a new generation of "green" cathode materials for rechargeable lithium batteries. We have developed nanocomposites combining graphene with two promising polymer cathode materials, poly(anthraquinonyl sulfide) and polyimide, to improve their high-rate performance. The polymer-graphene nanocomposites were synthesized through a simple in situ polymerization in the presence of graphene sheets. The highly dispersed graphene sheets in the nanocomposite drastically enhanced the electronic conductivity and allowed the electrochemical activity of the polymer cathode to be efficiently utilized. This allows for ultrafast charging and discharging; the composite can deliver more than 100 mAh/g within just a few seconds.     

A review of robust design methods for multiple responses

Problems in engineering design often involve determining design variable settings to optimize individual product performance for multiple criteria, which are often in conflict. We review mathematically rigorous techniques from the statistical literature for finding a vector x of design variable settings, which produces an optimal compromise solution among a group of prioritized response variables. The best compromise solution is typically gained by optimizing an objective function, which incorporates the prioritized demands of multiple responses. Since most multi-response objective functions are constructed by combining the functions used to optimize univariate responses, a review of the prominent univariate approaches is presented first. A multivariate approach from the engineering literature called the compromise Decision Support Problem is also reviewed. Finally a table comparing the relative merits of the different multivariate approaches summarizes the article in a concise and user-friendly fashion.     

Fast imaging of hard x rays with a laboratory microscope

An improved x-ray microscope with a fully electronic CCD detector system has been constructed that allows improved laboratory-based microstructural investigations of materials with hard x rays. It uses the Kirkpatrick-Baez multilayer mirror design to form an image that has a demonstrated resolution of 4 mum at 8 keV (Cu K(alpha) radiation). This microscope performs well with standard sealed-tube laboratory x-ray sources, producing digital images with 20-s exposure times for a 5-mum Au grid (a thickness of two absorption lengths).     

Induction of long-term receptive field plasticity in the auditory cortex of the waking guinea pig by stimulation of the nucleus basalis.

Learning induces neuronal receptive field (RF) plasticity in primary auditory cortex. This plasticity constitutes physiological memory as it is associative, highly specific, discriminative, develops rapidly, and is retained indefinitely. This study examined whether pairing a tone with activation of the nucleus basalis (NB) could induce RF plasticity in the waking guinea pig and, if so, whether it could be retained for 24 hrs. Subjects received 40 trials of a single frequency paired with electrical stimulation of the NB at tone offset. The physiological effectiveness of NB stimulation was assessed later while subjects were anesthetized with urethane by noting whether stimulation produced cortical desynchronization. Subjects in which NB stimulation was effective did develop RF plasticity and this was retained for 24 hrs. Thus, activation of the NB during normal learning may be sufficient to induce enduring physiological memory in auditory cortex. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Vortex-induced flame extinction in two-phase counterflow diffusion flames with CH planar laser-induced fluorescence and particle-image velocimetry

Here the interaction between a laminar two-phase, non-pre-mixed counterflow flame and a vortex is examined. Special emphasis is given to the influence of different flame and vortex parameters on the extinction behavior of the flame. Simultaneous planar laser-induced fluorescence of the CH radical layer produced by the flame and particle-image velocimetry measurements of the flowfield are used to characterize the flame-vortex interaction. These simultaneous diagnostics are used for the first time in this configuration. The extinction processes occurring during the flame-vortex interaction can be analyzed by this method, especially the influence of strain at the flame surface. The influence of the droplets on the extinction behavior appears clearly compared with a fully gaseous flame. The spray flame is weaker and extinguishes earlier than does a gaseous flame. In the measurements an additional broadband signal in the vicinity of the CH layer is probably due to the induced fluorescence of polycyclic aromatic hydrocarbons, excited at the same wavelength.     

Opinicon II: The state of the discipline review.

Informs that since the autumn of 1981, a growing group of psychologists have been carefully considering the current state of the discipline of psychology in Canada. The goal is an invited conference that would reflect on the major issues affecting psychology today and that would make recommendations to the Canadian Psychological Association and, indeed, to all Canadian psychology regarding the steps to be taken in the future to enhance the growth and development of psychology as a science and as a profession. Earlier state of the discipline reviews are briefly described and the terms of reference for the current project, Opinicon II, are presented. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Happy affect and smiling: Is their relation moderated by interpersonal power?

Three studies examined the hypothesis that the relation between self-rated happy affect and overall smiling during dyadic interactions is moderated by one's relative power. In all 3 studies, interpersonal power was experimentally manipulated, and smiling and self-reported happy affect were measured during and after the interaction, respectively. Happy affect was positively correlated with overall smiling for both low-power and high-power participants. Moreover, the correlations for both low- and high-power participants were comparable to those found when power roles were not unequal or made salient, based on analysis of the present studies as well as comparison with the previously published literature. Happy affect and overall smiling had a positive relation that was not moderated by interpersonal power. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Access and service convergence over the mobile internet – A survey

Next generation heterogeneous wireless networks are expected to interwork with Internet Protocol (IP)-based infrastructures. Conventional network services operate like silos in that a specific set of services are offered over a specific type of access network. It is desirable for users to be able to roam between fixed and mobile networks that employ different access technologies. Therefore, mobility management with quality of service (QoS) support is of particular importance and one of the driving forces of convergence. Since service providers often provide more than one service to their subscribers, it is important to facilitate convergence of network charging architecture through a common charging framework. One of the main issues of IP-based convergence is security and privacy. This requires coordination of different security policies in diverse networks that have different security levels and capabilities. The business case for migration to an IP-based platform motivates operators to deliver more powerful services for customers as well as a better user experience. This paper provides an overview of converged mobile Internet architectures and their implications on QoS, charging/billing and security, as well as emerging business models for telecommunication services.