Amide Neighbouring‐Group Effects in Peptides: Phenylalanine as Relay Amino Acid in Long‐Distance Electron Transfer

In nature, proteins serve as media for long‐distance electron transfer (ET) to carry out redox reactions in distant compartments. This ET occurs either by a single‐step superexchange or through a multi‐step charge hopping process, which uses side chains of amino acids as stepping stones. In this study we demonstrate that Phe can act as a relay amino acid for long‐distance electron hole transfer through peptides. The considerably increased susceptibility of the aromatic ring to oxidation is caused by the lone pairs of neighbouring amide carbonyl groups, which stabilise the Phe radical cation. This neighbouring‐amide‐group effect helps improve understanding of the mechanism of extracellular electron transfer through conductive protein filaments (pili) of anaerobic bacteria during mineral respiration.     

Density doubling of block copolymer templated features

Block copolymers can be used to template large arrays of nanopatterns with periodicities equal to the characteristic spacing of the polymer. Here we demonstrate a technique capitalizing on the multilayered arrangement of cylindrical domains to effectively double the pattern density templated by a given polymer. By controlling the initial thickness of the film and the solvent annealing conditions, it was possible to reproducibly create density doubled lines by swelling the film with solvent until bilayers of horizontal cylinders were obtained. This process was also demonstrated to be compatible with graphoepitaxy.     

A Proteomic Approach of Bradyrhizobium/Aeschynomene Root and Stem Symbioses Reveals the Importance of the fixA Locus for Symbiosis

Rhizobia are soil bacteria that are able to form symbiosis with plant hosts of the legume family. These associations result in the formation of organs, called nodules in which bacteria fix atmospheric nitrogen to the benefit of the plant. Most of our knowledge on the metabolism and the physiology of the bacteria during symbiosis derives from studying roots nodules of terrestrial plants. Here we used a proteomics approach to investigate the bacterial physiology of photosynthetic Bradyrhizobium sp. ORS278 during the symbiotic process with the semi aquatical plant Aeschynomene indica that forms root and stem nodules. We analyzed the proteomes of bacteria extracted from each type of nodule. First, we analyzed the bacteroid proteome at two different time points and found only minor variation between the bacterial proteomes of 2-week- and 3-week-old nodules. High conservation of the bacteroid proteome was also found when comparing stem nodules and root nodules. Among the stem nodule specific proteins were those related to the phototrophic ability of Bradyrhizobium sp. ORS278. Furthermore, we compared our data with those obtained during an extensive genetic screen previously published. The symbiotic role of four candidate genes which corresponding proteins were found massively produced in the nodules but not identified during this screening was examined. Mutant analysis suggested that in addition to the EtfAB system, the fixA locus is required for symbiotic efficiency.     

Process analysis of adult age effects on a computer-administered Trail Making Test.

Two studies were conducted in which healthy adults of different ages performed computer-administered tests resembling the R. M. Reitan (1992) Trail Making Test. In both studies, increased age was associated with slower performance, but there were no age differences in the efficiency of the path used to connect the targets. Analyses of independent age-related influences revealed that most of the age differences in trail-making performance were attributable to a slower speed of executing relevant operations. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Examining the Role of Fire Prevention on Hoarding Response Teams: Vancouver Fire and Rescue Services as a Case Study

Hoarded homes present a significant threat to the safety of occupants of the home and adjacent homes as well as emergency responders. Extreme accumulation of combustible materials raises the fuel load and prevents egress. Despite having regulatory tools related to fire prevention, fire departments generally lack the necessary background to address the many challenges related to the complex mental health problem of hoarding. Using Vancouver as a case study, this paper highlights the role of Vancouver Fire and Rescue Services in the City’s coordinated hoarding response and outlines their fire code inspections protocol developed for use in hoarding cases. On the basis of fire inspection reports, we compare the processes and outcomes of complaint-driven hoarding inspections with standard annual fire inspections of restaurants. Hoarding cases take longer to resolve and are more challenging, but inspectors achieved satisfactory status in 94% of cases without prosecution. About 20% of hoarding complaints do not involve fire code violations. Although this approach requires an investment and engagement with other community agencies, it represents a useful model for other jurisdictions.     

Controlled growth and investigations on the morphology and mechanical properties of hydroxyapatite/titania nanocomposite thin films

In this paper, the focus is on understanding the properties of nanocomposite hydroxyapatite (HAp)/titania (TiO₂) thin films with respect to TiO₂ concentration. HAp/TiO₂ nanostructured composite thin films with different TiO₂ concentrations were successfully fabricated by a simple sol–gel dip coating method. Highly stable HAp and TiO₂ sols were prepared prior to the formation of nanocomposite thin films. The coatings were performed under controlled dipping and heat treatment processes. Phase pure HAp and TiO₂ were well developed in the nanocomposite after the heat treatment and this was confirmed by XRD. The SEM and AFM analyses of HAp/TiO₂ nanocomposite coatings show the variation in the morphology as a consequence different TiO₂ concentration. This shows a reduction in the particle size to nanoscale due to the addition of TiO₂. The mechanical strength of the coating also increased upon the addition of TiO₂ as determined by nanoindentation. The composite thin films with 50 and 80 vol.% of TiO₂ show good mechanical strength when compared to other concentrations of TiO₂.     

Nanostructured leaf like hydroxyapatite/TiO2 composite coatings by simple sol-gel method

We demonstrate an approach for the coating of nanostructured leaf like hydroxyapatite (HAp)/TiO₂ composite on glass substrate by sol-gel dip coating process. HAp/TiO₂ nanocomposite thin film was obtained by controlling the dipping rate and the dipping cycle. It was observed from Scanning Electron Microscope that leaf like nanostructured film was deposited on the glass substrate. However, the structure of the film was changed with the dipping cycle and dipping rate. Transmission Electron Microscopic analysis further confirms the morphology of the nanostructured coatings. The presence of Ti, O, Ca and P was detected by Energy Dispersive X-ray Analysis. We further confirmed the composite by X-ray diffraction analysis. Atomic Force Microscope analysis indicates that the films are composed of nanoparticles ranging from 100 to 200 nm and the films were observed to present well-defined grain boundaries. It has been shown that nanocomposite coatings are dependent on the sol concentration, dipping parameters, and the composition of HAp and TiO₂.     

Monolithic column HPLC separation of intact proteins analyzed by LC-MALDI using on-plate digestion: An approach to integrate protein separation and identification

A method is developed to integrate a protein separation by monolithic capillary reversed-phase high-performance liquid chromatography to on-probe tryptic digestion for subsequent analyses by MALDI-TOF MS and MALDI-TOF/TOF MS. The method provides a means of directly interfacing separations to MALDI-MS, reducing the amount of time required for traditional procedures involving in-solution enzymatic digestion and sample cleanup prior to MALDI-MS analysis. When used with pI-based fractionation as a first dimension, it provides a means of analyzing complex mixtures of proteins with minimal sample handling and cleanup. The use of monolithic capillary columns sufficiently resolved intact proteins so that peptide mass fingerprinting analysis by MALDI-TOF MS resulted in the identification of close to 40 unique proteins from 120 ng of sample obtained from a prefractionated MCF10 cell line at pH 6.34, where the identifications of several of these proteins were also confirmed by intact MW and tandem mass spectrometric analysis. The reproducibility of this method has been demonstrated to be sufficient for the purpose of protein identifications. Experimental values of protein intact MW are obtained and compared to that expected for each protein identified.     

Ion mobility-mass spectrometry (IM-MS) for top-down proteomics: increased dynamic range affords increased sequence coverage

A general approach that combines mass spectrometry (MS), collision-induced dissociation (CID), ion mobility (IM), and MS for top-down proteomics is described, denoted as MS-CID-IM-MS. Using this approach, CID product ions are dispersed in two dimensions, specifically size-to-charge (IM) and mass-to-charge (MS), and the resulting 2D data display greatly facilitates peptide/protein mass mapping, amino acid sequence analysis, and determination of site-specific protein modifications. Also, this approach alleviates some of the inherent limitations of top-down proteomics, viz. the limitations in dynamic range for fragment ion abundances owing to the number of fragmentation channels available to large ionic systems as well as the resulting spectral congestion. For large peptides such as melittin (2845 Da), CID of the [M + 3H](3+), [M + 4H](4+), and [M + 5H](5+) ions yields amino acid sequence coverage of 42.3%, 38.5%, and 7.7%, respectively, whereas the hybrid MS-CID-IM-MS approach yields amino acid sequence coverages of 84.6%, 65.4%, and 69.2%, respectively. For large biomolecules such as ubiquitin (8565 Da), the amino acid sequence coverage increases from 39% to 76%. The MS-CID-IM-MS top-down approach allows for greater depth of information by allowing the assignment and study of internal fragment ions. Lastly, analysis of the methyl esterification of ubiquitin and single point mutation of human iron sulfur cluster U (HISCU, 14.3 kDa) demonstrates the ability of MS-CID-IM-MS to rapidly identify the presence and sites of modifications.