Determination of ligand-protein dissociation constants by electrospray mass spectrometry-based diffusion measurements

A novel approach for the quantification of ligand-protein interactions is presented. Electrospray ionization mass spectrometry (ESI-MS) is used to monitor the diffusion behavior of noncovalent ligands in the presence of their protein receptors. These data allow the fraction of free ligand in solution to be determined, such that the corresponding dissociation constants can be calculated. A set of conditions is developed that provides an "allowable range" of concentrations for this type of assay. The method is tested by applying it to two different inhibitor-enzyme systems. The dissociation constants measured for benzamidine-trypsin and for N,N',N' '-triacetylchitotriose-lysozyme are (50 +/- 10) and (6 +/- 1) mM, respectively. Both of these results are in good agreement with previous data from the literature. In contrast to traditional ESI-MS-based methods, the approach used in this work does not rely on the preservation of specific solution-type noncovalent interactions in the gas phase. It is shown that this method allows an accurate determination of dissociation constants, even in cases in which the ion abundance ratio of free to ligand-bound protein in ESI-MS does not reflect the corresponding concentration ratio in solution.     

Learning to recognize talkers from natural, sinewave, and reversed speech samples.

In 5 experiments, the authors investigated how listeners learn to recognize unfamiliar talkers and how experience with specific utterances generalizes to novel instances. Listeners were trained over several days to identify 10 talkers from natural, sinewave, or reversed speech sentences. The sinewave signals preserved phonetic and some suprasegmental properties while eliminating natural vocal quality. In contrast, the reversed speech signals preserved vocal quality while distorting temporally based phonetic properties. The training results indicate that listeners learned to identify talkers even from acoustic signals lacking natural vocal quality. Generalization performance varied across the different signals and depended on the salience of phonetic information. The results suggest similarities in the phonetic attributes underlying talker recognition and phonetic perception. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

New anionic surface‐active agent derived from wool proteins for hair treatment

An anionic surface‐active agent derived from wool proteins was developed as an alternative to the anionic surfactants extensively used in shampoo formulations. The physicochemical properties of this new surface‐active agent prepared form wool proteins and its application for human hair treatments were studied. This new product could be considered a new mild anionic surface‐active agent, as evidenced by the results found by the evaluation of its physicochemical properties. The new wool anionic surface‐active agent was shown to be very substantive to hair, coating the fiber surface, giving rise to a significant improvement in the mechanical properties of the hair fibers, and providing a certain damage‐prevention effect on the hair. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2010     

Effect of kafirin protein coating on sensory quality and shelf-life of 'Packham's Triumph' pears during ripening

BACKGROUND: Pears are exported in large quantities from South Africa, resulting in large revenues. Minimisation of quality losses once the fruit has reached the export destination is as important as following strict export and distribution protocols. Kafirin can form edible films. In this study an edible coating made from 20 g kg^?1 kafirin coating solution was applied as a postharvest treatment to retard quality deterioration of ‘Packham's Triumph’ pears during storage at the typical ripening temperature (20 °C). Changes in physicochemical and sensory quality were monitored over a period of 24 days. RESULTS: The kafirin coating was unable to retard the onset of ripening but decreased the respiration rate and retarded the progression of senescence. However, moisture loss was exacerbated in the kafirin‐coated fruit during ripening at 20 °C, especially towards the end of the shelf‐life. CONCLUSION: The kafirin coating extended the eat‐ripe quality of the pears by 1–2 weeks. However, the appearance of the fruit was unacceptable after 14 days of storage in terms of wrinkled skin. Further work is needed to improve the water barrier properties of the kafirin coating by incorporating a wax or triglyceride into the coating formulation or more simply by applying a kafirin coating to waxed fruit. Copyright © 2011 Society of Chemical Industry     

A localised experimental–numerical technique for determining mixed mode strain energy release rates

Composite structures are being used increasingly in the aerospace industry due to their superior specific stiffness and strength. One key issue associated with such structures is delamination, and how to effectively predict this. A new method which is derived from localised test displacement data is presented to determine the mixed mode strain energy release rates of layered structures with a pre-existing crack. Images taken during experimentation of the vicinity of the crack tip are analysed at low load and high load to determine the displacement changes across the load variation. These displacements are applied as boundary conditions to a simple local numerical model including a constraint at the crack tip. The forces and displacements at the crack tip are taken as output data and combined with the Virtual Crack Closure Technique to predict strain energy release rates. Initial validation of the localised experimental–numerical technique (LENT) shows that applying experimental data to a numerical model does give reasonable agreement thus far in the established trends, and hence LENT is promising for use in determining mixed mode strain energy release rates and mode mixity ratios.     

Mechanism of Crystallization and Implications for Charge Transport in Poly(3‐ethylhexylthiophene) Thin Films

In this work, crystallization kinetics and aggregate growth of poly(3‐ethylhexylthiophene) (P3EHT) thin films are studied as a function of film thickness. X‐ray diffraction and optical absorption show that individual aggregates and crystallites grow anisotropically and mostly along only two packing directions: the alkyl stacking and the polymer chain backbone direction. Further, it is also determined that crystallization kinetics is limited by the reorganization of polymer chains and depends strongly on the film thickness and average molecular weight. Time‐dependent, field‐effect hole mobilities in thin films reveal a percolation threshold for both low and high molecular weight P3EHT. Structural analysis reveals that charge percolation requires bridged aggregates separated by a distance of ≈2–3 nm, which is on the order of the polymer persistence length. These results thus highlight the importance of tie molecules and inter‐aggregate distance in supporting charge percolation in semiconducting polymer thin films. The study as a whole also demonstrates that P3EHT is an ideal model system for polythiophenes and should prove to be useful for future investigations into crystallization kinetics.     

Ligand-like effects induced by anti-c-erbB-2 antibodies do not correlate with and are not required for growth inhibition of human carcinoma cells

The c-erbB-2 gene encodes a M(r) 185,000 tyrosine kinase receptor (p185) with extensive homology to the epidermal growth factor receptor. We have conducted mechanistic studies with several anti-p185 monoclonal antibodies (TAb 250, -255, -257, -260, and -263) directed against the extracellular domain of p185 utilizing the SKBR-3, BT-474, and SKOV-3 cancer cell lines. Several of these antibodies exhibited ligand-mimicking properties: they induced tyrosine phosphorylation of p185; increased the catalytic activity of the receptor substrate phospholipase C-gamma 1; exhibited time- and pH-dependent internalization; induced receptor down-regulation; and increased the turnover of the p185 protein delta 3-fold. However, there was not a universal correlation between the antibody-mediated ligand-like effects and growth inhibition. TAb 250 inhibited BT-474 cells but did not alter p185 phosphotyrosine content or increase receptor turnover in these cells. TAb 260 increased p185 protein turnover but did not affect proliferation of the SKOV-3 cell line. Furthermore, blockade of TAb 250-induced receptor phosphorylation with the tyrosine kinase inhibitor tyrphostin 50864-2 did not abrogate TAb 250-mediated growth inhibition of SKBR-3 cells. These data suggest that ligand-like effects mediated by p185 antibodies are not critical for the growth inhibition of c-erbB-2-overexpressing carcinoma cells.     

Degradation of Polycarbonate Properties Under Thermal Aging

Journal of Failure Analysis and Prevention - The aim of this paper is to characterize the thermal aging effect near the glass transition temperature of polycarbonate (PC) at different hold time....     

Ammonium Formate-Pd/C as a New Reducing System for 1,2,4-Oxadiazoles. Synthesis of Guanidine Derivatives and Reductive Rearrangement to Quinazolin-4-Ones with Potential Anti-Diabetic Activity

1,2,4-Oxadiazole is a heterocycle with wide reactivity and many useful applications. The reactive O-N bond is usually reduced using molecular hydrogen to obtain amidine derivatives. NH₄CO₂H-Pd/C is here demonstrated as a new system for the O-N reduction, allowing us to obtain differently substituted acylamidine, acylguanidine and diacylguanidine derivatives. The proposed system is also effective for the achievement of a reductive rearrangement of 5-(2′-aminophenyl)-1,2,4-oxadiazoles into 1-alkylquinazolin-4(1H)-ones. The alkaloid glycosine was also obtained with this method. The obtained compounds were preliminarily tested for their biological activity in terms of their cytotoxicity, induced oxidative stress, α-glucosidase and DPP4 inhibition, showing potential application as anti-diabetics.