Stereopositional Outcome in the Packing of Dissimilar Aromatics in Designed β‐Hairpins

A popular strategy in the de novo design of stable β‐sheet structures for various biomedical applications is the incorporation of aromatic pairs at the non‐hydrogen‐bonding (NHB) position. However, it is important to explicitly understand how aryl pair packing at the NHB region is coordinated with backbone structural rearrangements, and to delineate the benefits and drawbacks associated with stereopositional choice of dissimilar aromatic pairs. Here, we probe the consequences of flipped Trp/Tyr pairs by using engineered permutants at the NHB position of dodecapeptide β‐hairpins, proximal and distal to the turn. Extensive conformational analysis of these peptides using NMR and CD spectroscopy reveal that a classic Edge‐to‐Face and Face‐to‐Edge geometry at the proximal and distal aromatic pairs, respectively, in YW‐WY, is the most stabilizing. Such a preferred packing geometry in YW‐WY results in a highly twisted β‐sheet backbone, with Trp always providing a ‘Face’ orientation to its dissimilar aromatic partner Tyr. Flipping the proximal and/or distal aromatic pair distorts the ideal T‐shaped geometry, and results in alternate aryl arrangements that can adversely affect strand twist and β‐sheet stability. Our study reveals the existence of a strong stereopositional influence on the packing of dissimilar aromatic pairs. Our findings highlight the importance of modeling physical interaction forces while designing protein and peptide structures for functional applications.     

Calix Receptor Edifice; Scrupulous Turn Off Fluorescent Sensor for Fe(III), Co(II) and Cu(II)

Novel Supramolecular fluorescence receptor derived from calix-system i.e. calix[4]resorcinarene bearing dansylchloride as fluorophore was designed and synthesized. The compound was purified by column chromatography and characterized by elemental analysis, NMR and Mass spectroscopy. Tetradansylated calix[4] resorcinarene (TDCR) shows a boat conformation with C₂v symmetry. The complexation behaviour of metal cations [Ag(I), Cd(II), Co(II), Fe(III), Hg(II), Cu(II), Pb(II), Zn(II), U(VI) (1?×?10^-4?M)] with tetra dansylated calix[4]resorcinarene (1?×?10^-6?M) was studied by spectophotometry and spectrofluorometry. Red shift in the absorption spectra led us to conclude that there is strong complexation Fe(III), Co(II) and Cu(II) with TDCR. These metal cations also produce quenching with red shifts in the emission spectra. The maximum quenching in emission intensity was observed in the case of Fe(III) and its binding constant was also found to be significantly higher than that of Co(II) and Cu(II). Quantum yield of metal complexes of Fe(III) was found to be lower in comparison with Co(II) and Cu(II) complexes. Stern Volmer analysis indicates that the mechanism of fluorescence quenching is either purely dynamic, or purely static.     

An inter-company cross-validation exercise on capillary electrophoresis testing of dose uniformity of paracetamol content in formulations

Summary Successful transfer of an MECC method for the determination of paracetamol between 7 companies is shown. The method was applied to the quantitative determination of paracetamol levels in capsules. The results obtained agreed well between companies and with both HPLC data and the label claim. Acceptable method performance in terms of relative migration time precision and response factor precision was reported by each company.     

Coumarin 6 and 1,6-diphenyl-1,3,5-hexatriene (DPH) as fluorescent probes to monitor protein aggregation

We report the use of Coumarin 6 and 1,6-diphenyl-1,3,5-hexatriene (DPH) for the identification of protein aggregates for the first time. The two dyes can be used at very low (nanomolar) concentrations and do not interfere with the aggregation process, as is reported for other commonly used fluorescent protein probes. In the presence of protein aggregates, their quantum yields are significantly high. DPH is able to recognize both amorphous and fibrillar aggregates but cannot distinguish between them. Coumarin 6 can distinguish between both types of aggregates. It also exhibits the characteristic sigmoidal curve of amyloid formation, with higher sensitivity for detection of fibrillation than the conventionally used Thioflavin T.     

Inter-company cross validation exercise on capillary electrophoresis. Quantitative determination of drug counter-ion level

Summary A capillary electrophoresis (CE) method for the quantitative determination of sodium levels in the sodium salt of an acidic drug has been transferred across 6 independent pharmaceutical companies. All companies were able to demonstrate similar selectivity and acceptable precision for migration times and relative migration times. Five companies obtained acceptable precision for the repeatability of response factors from repeated injections of calibration solutions. Highly variable results from the sixth company indicated problems arising from contamination from residual sodium, possibly from unclean glassware or autosampler vials.The quantitative assay results from all companies confirmed the correct drug: counter-ion ratio. The average % w/w sodium content was 5.45% which represents 99.0% of the theoretical sodium content of sodium cephalothin     

Structural and electrical properties of sol–gel grown (1 − x) (ZnO) + (x) (SnO2) (x = 0, 0.25, 0.5) nanocomposites

Journal of Sol-Gel Science and Technology - Semiconducting oxide nanocomposites of ZnO/SnO2 with different weight ratio, i.e. (i) ZnO:SnO2?=?100:0 (ZnO0), (ii)...