Capillary electrophoresis of proteins 2005-2007

This review article with 239 references describes recent developments in capillary electrophoresis of proteins, and covers the two years since the previous review (V. Dolník, Electrophoresis 2006, 27, 126-141) through spring 2007. It includes topics related to CE of proteins, such as sample pretreatment, wall coatings, improving separation, various forms of detection, and special electrophoretic techniques including ACE, CIEF, capillary ITP, and CEC. The paper describes applications of CE to analysis of proteins in real-world samples including human body fluids, food and agricultural samples, protein pharmaceuticals and recombinant protein preparations.     

Photoswitchable Phosphonate–Fullerene Hybrids with Cholinesterase Activity

Modern progress in photopharmocology calls for new generation of compounds joining bioactivity, photoswitchable properties and high selectivity of response to light wavelength. Introduced here, phosphonate–fullerene hybrids are the first representatives of such compounds. Phosphonate–fullerene hybrids were synthesized on a base of fullerene C₆₀ and organophosphates with the function of photoswitchable cholinesterase activity—phosphorylated thiazolotriazole and aminomalonate compounds and studied with FTIR, UV–VIS spectroscopy and IPC-micro neurotoxin amperometric analysis. As a result of spectroscopic and bioactivity characterization, it was not only demonstrated butyrylcholinesterase (BuChE) inhibition increase in phosphonate–fullerene hybrids compared with pure phosphonates but also pronounced response of inhibition degree to laser irradiation of hybrids. It was found opposite behavior of hybrids as a result of laser irradiation—BuChE inhibition drop-off for thiazolotriazole–fullerene and pronounced growth for aminomalonate–fullerene. The other remarkable peculiarity of presented phosphonate–fullerene hybrids is high selectivity of inhibition change degree to laser wavelength (266 or 325 nm).     

Effect of π-Extended Substituents on Photophysical Properties of BODIPY Dyes in Solutions

Four boron-dipyrrine (BODIPY) based dyes with π-extended substituents in 8-position of dipyrrin ligand have been synthesized and characterized. Photophysical properties of the obtained compounds have been investigated in different individual solvents. Deposits of solvent polarity and viscosity were evaluated. BODIPY with 8-biphenyl substituent was found to be the fluorescent molecular rotor in contrast to more extended substituents. The complex nature of solvent-solute interactions leads to the poor applicability of standard multiparameter approaches to BODIPY solvatochromic properties. Fluorescence intensity was found to increase in case of solvent polarity growth, it is not typical for BODIPY. Taking that into account the BODIPY with π-extended substituents could be used for fluorescence viscosity measurements, and as the fluorescent media polarity indicators in analytical chemistry and biochemistry.     

New exact solutions of the massive Dirac equation with electric or scalar potential

A new class of exact solutions for the massive Dirac equation with electric or scalar potential depending on one independent variable is constructed. This construction is based on quaternionic reformulation of the Dirac equation. Copyright © 2000 John Wiley & Sons, Ltd.     

Reactions of dithioxo-1,3,2λ5,4λ5-dithiadiphosphetanes with triorganolead derivatives

The reactions of Lawesson's reagent 1a and its 4-ethoxy homologue 1b with triethyl- and triphenyl(alkoxy)plumbanes 2a,b and -(alkylthio)plumbanes 4a,b were studied. On the basis of these reactions, novel, advantageous methods of synthesizing S-triethyl and triphenylplumbyl derivatives of aryldithio- and trithiophosphonic acids 3a–d and 5a,b were developed. © 1997 John Wiley & Sons, Inc.     

Twice as Nice: The Duff Formylation of Umbelliferone Revised

More efficient and preferably more convenient and greener synthetic solutions in coumarin scaffold functionalization are in steady demand. The Duff ortho-formylation of unsubstituted umbelliferone was revised in this study. The reaction conditions were optimized based upon data from the literature analysis and resulted in unexpectedly rapid ortho-formylation of umbelliferone, yielding a mixture of ortho-formyl position isomers. Thorough studies on the separation of ortho-formylated umbelliferones using chromatographic and recrystallization methods as well as the evaluation of their solubility in common organic solvents led to complete resolution of 8-formyl- and 6-formylumbelliferones. The precise protocol for simultaneous preparation, extraction, and purification of 8-formyl- and 6-formylumbelliferones is provided, and the prospective studies of biological and pharmacological activities of these compounds are synopsized.     

Analytical Expressions for Ising Models on High Dimensional Lattices

We use an m-vicinity method to examine Ising models on hypercube lattices of high dimensions d≥3. This method is applicable for both short-range and long-range interactions. We introduce a small parameter, which determines whether the method can be used when calculating the free energy. When we account for interaction with the nearest neighbors only, the value of this parameter depends on the dimension of the lattice d. We obtain an expression for the critical temperature in terms of the interaction constants that is in a good agreement with the results of computer simulations. For d=5,6,7, our theoretical estimates match the numerical results both qualitatively and quantitatively. For d=3,4, our method is sufficiently accurate for the calculation of the critical temperatures; however, it predicts a finite jump of the heat capacity at the critical point. In the case of the three-dimensional lattice (d=3), this contradicts the commonly accepted ideas of the type of the singularity at the critical point. For the four-dimensional lattice (d=4), the character of the singularity is under current discussion. For the dimensions d=1, 2 the m-vicinity method is not applicable.     

Analysis of optical characteristics of holey photonic crystal devices with the extended coupled-mode formalism

Presented here is a new approach for analysis of the so-called holey photonic crystals—a class of electro-optical components, in which periodicity of air holes in dielectric media is used for confinement of light. This class includes several kinds of microstructured fibers, semiconductor lasers etc. Accurate evaluation of optical characteristics of those devices is usually a complicated problem due to the large dimensions and the fine structure of their refractive index distribution. Furthermore, usually, only numerical solutions for this class of optical components are available. The overwhelming majority of the physical models, suitable for analysis of holey photonic devices, proceed from the “natural” assumption: the devices are considered as arrays of air holes, surrounded by dielectric material. In this work we propose another model. Namely, we treat them as arrays of dielectric spots (waveguides), embedded in the air (cladding material). This model allows utilization of the extended coupled-mode theory (a relatively new approach designed for analysis of infinite arrays of coupled waveguides and previously considered inapplicable to holey optical components) for calculations of the latter. In this sense, we present a new method for analysis of holey photonic crystals. On the one hand, our method allows analytical evaluation of some optical characteristics of holey optical components (such as the number of photonic bands and bandwidth). On the other hand, accurate numerical computation of the photonic band structure of the holey photonic devices, incorporating a large number of holes, can be done with this technique on a timescale of several minutes.     

Short monolithic columns for purification and fractionation of peptide samples for matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry analysis in proteomics

This study records a novel application of methacrylate-based monolithic columns for MALDI-TOF/TOF MS analyses in proteomics for pre-concentration and separation of peptides derived from protein digestion. Reversed-phase monolithic capillary columns (30 mm × 0.32 mm i.d.) were created inside the fused silica capillary via thermal-initiated free-radical polymerization of ethylene glycol dimethacrylate and lauryl methacrylate monomers in the presence of 1-propanol and 1,4-butandiol as a porogen system. The elution of peptides was achieved using a linear gradient of acetonitrile from 0 to 60% in water with 0.1% trifluoroacetic acid formed in a microsyringe. Individual fractions of separated peptides were collected on the MALDI target spots covered with alpha-cyano-4-hydroxycinnamic acid used as a matrix and then they were analyzed using MALDI-TOF/TOF mass spectrometry. The developed method was tested with a mixture of tryptic peptides from bovine serum albumin and its applicability was also tested for tryptic in-gel digests from barley grain extracts of water soluble proteins separated using SDS gel electrophoresis. The number of detected peptides was approximately three to four times higher compared to the analysis without previous separation. These results show an improved quality of sample information with the higher amount of identified peptides which increased protein sequence coverage and improved sensitivity of mass spectrometry measurements.     

Shape- and Size-Selective Preparation of Hectorite-Supported Ruthenium Nanoparticles for the Catalytic Hydrogenation of Benzene

The cationic organometallic aqua complexes formed by hydrolysis of [(C₆H₆)₂RuCl₂]₂ in water, mainly [(C₆H₆)Ru(H₂O)₃]²⁺, intercalate into white sodium hectorite, replacing the sodium cations between the anionic silicate layers. The yellow hectorite thus obtained reacts in water with molecular hydrogen (50 bar, 100 °C) to give a dark suspension containing a black hectorite in which large hexagonally shaped ruthenium nanoparticles (20–50 nm) are intercalated between the anionic silicate layers, the charges of which being balanced by hydronium cations. If the reduction with molecular hydrogen (50 bar, 100 °C) is carried out in various alcohols, spherical ruthenium nanoparticles of smaller size (3–38 nm depending on the alcohol) are obtained. In alcohols other than methanol, the reduction also works without H₂ under reflux conditions, the alcohol itself being the reducing agent; the ruthenium nanoparticles obtained in this case are spherical and small (2–9 nm) but tend to aggregate to form clusters of nanoparticles. Whereas the ruthenium nanoparticles prepared by reduction of the yellow hectorite in refluxing alcohols without hydrogen pressure are almost inactive, the nanoparticles formed by hydrogen reduction catalyze the hydrogenation of benzene to give cyclohexane under mild conditions (50 °C) with turnover frequencies up to 6500 catalytic cycles per hour, the best solvent being ethanol. Dedicated to Professor C. N. R. Rao, pioneer of nanocluster chemistry, on the occasion of his 75th birthday.