Optimization of protein sample preparation for two-dimensional electrophoresis

Optimized 2DE sample preparation protocols that maximize total protein recovery are fundamental to improving proteome capture and increasing the utility of 2DE, which is in part limited by inadequate recovery of proteins with diverse physicochemical properties. Maintaining protein solubility is an important factor for protein recovery, but the multitude of solubility-enhancing agents and the relatively low-throughput nature of 2DE limit the systematic study of sample preparation. In this work, design of experiment (DOE) approaches are used to optimize protein recovery by altering the levels of four solubility-enhancing agents (urea, DTT, CHAPS, and SDS) in the initial suspension solution. Protein recovery is quantified by a total protein concentration assay, which is demonstrated to be representative of SDS-PAGE and 2DE recovery. DOE methodologies are presented as relatively high-throughput procedures for optimizing 2DE sample preparation parameters for a variety of sample types. Optimal suspension solution compositions are shown to vary across a model protein solution (no urea or DTT), Chinese hamster ovary (CHO) cell lysate (8 M urea, ≥2% CHAPS, ≥32.5 mM DTT), and Escherichia coli cell lysate (8 M urea, 4% CHAPS, 65 mM DTT), with optimized conditions increasing 2DE protein recovery at least 50% compared to suboptimal conditions.     

Microwave-assisted Claisen rearrangement of naphthyl 2-propynyl ethers: synthesis of naphthofurans

An efficient two-step approach for the synthesis of naphtho[1,2-b]furans and naphtho[2,1-b]furans has been developed. Various functionalized propargyl alcohols were etherified with α- or β-naphthol under Mitsunobu reaction conditions to give naphthyl 2-propynyl ethers, which underwent a facile microwave-assisted Claisen rearrangement and concomitant anionic cyclization to yield naphthofuran derivatives under basic reaction conditions.     

Organic–inorganic tandem route to polymer nanocomposites: kinetic products versus thermodynamic products

We report a facile single-step photochemical methodology to afford alkylsiloxane-polymer hybrid films which relies on the tandem photoacid-catalyzed polymerization of n-alkyltrimethoxysilane precursors and a diglycidyl ether organic monomer. Photoacids liberated by the UV decomposition of iodonium salt triggers simultaneously the sol–gel process and epoxy cationic polymerization. Such conditions are intended to favour the formation of kinetic products trapped by cross-linking reactions instead of the thermodynamically most stable structures, whose preference would be for macrophase segregation. Organosilane precursor exhibiting different structures and chain lengths (n-butyl, n-octyl, n-dodecyl, n-hexadecyl, isobutyl and isooctyl) were systematically investigated to afford a range of transparent alkylsiloxane-polyether hybrids. The competitive organic–inorganic reaction kinetics were investigated using in situ real-time Fourier transform infrared spectroscopy. A main emphasis has been on discussing the effect of the alkyl substituent structure on the photoinduced polymerization kinetics and the silicate networks characterized by ²⁹Si solid-state NMR. To avoid phase separation, the rate of formation of the two phases was tailored to favour concomitancy upon modulating several experimental parameters: film thickness, alkyl structure, photoacid generator concentration. Finally, the viscoelastic and surface properties were also assessed by dynamic mechanical analysis and water contact angle measurements, respectively.     

Synthesis,Structural Characterization,Photophysics, and Broadband Nonlinear Absorption of a Platinum(II) Complex with the 6‐(7‐Benzothiazol‐2′‐yl‐9,9‐diethyl‐9 H‐fluoren‐2‐yl)‐2,2′‐bipyridinyl Ligand

A platinum complex with the 6‐(7‐benzothiazol‐2′‐yl‐9,9‐diethyl‐9H‐fluoren‐2‐yl)‐2,2′‐bipyridinyl ligand ( 1 ) was synthesized and the crystal structure was determined. UV/Vis absorption, emission, and transient difference absorption of 1 were systematically investigated. DFT calculations were carried out on 1 to characterize the electronic ground state and aid in the understanding of the nature of low‐lying excited electronic states. Complex 1 exhibits intense structured ¹π–π* absorption at λ_abs<440 nm, and a broad, moderate ¹M LCT/¹LLCT transition at 440–520 nm in CH₂Cl₂ solution. A structured ³π–π*/³M LCT emission at about 590 nm was observed at room temperature and at 77 K. Complex 1 exhibits both singlet and triplet excited‐state absorption from 450 nm to 750 nm, which are tentatively attributed to the ¹π–π* and ³π–π* excited states of the 6‐(7‐benzothiazol‐2′‐yl‐9,9‐diethyl‐9H‐fluoren‐2‐yl)‐2,2′‐bipyridine ligand, respectively. Z‐scan experiments were conducted by using ns and ps pulses at 532 nm, and ps pulses at a variety of visible and near‐IR wavelengths. The experimental data were fitted by a five‐level model by using the excited‐state parameters obtained from the photophysical study to deduce the effective singlet and triplet excited‐state absorption cross sections in the visible spectral region and the effective two‐photon absorption cross sections in the near‐IR region. Our results demonstrate that 1 possesses large ratios of excited‐state absorption cross sections relative to that of the ground‐state in the visible spectral region; this results in a remarkable degree of reverse saturable absorption from 1 in CH₂Cl₂ solution illuminated by ns laser pulses at 532 nm. The two‐photon absorption cross sections in the near‐IR region for 1 are among the largest values reported for platinum complexes. Therefore, 1 is an excellent, broadband, nonlinear absorbing material that exhibits strong reverse saturable absorption in the visible spectral region and large two‐photon‐assisted excited‐state absorption in the near‐IR region.     

Selective and Sensitive Electrochemical Sensor for L‐Methionine at Physiological pH Using Functionalized Triazole Polymer Film Modified Electrode

This communication describes the determination of an essential amino acid, L ‐methionine (L ‐Met) in the presence of important interferents, ascorbic acid (AA) and uric acid (UA) at physiological pH using a glassy carbon electrode modified with an electropolymerized film of 3‐amino‐5‐mercapto‐1,2,4‐triazole (p‐AMTa). The bare glassy carbon electrode fails to show a voltammetric signal for L ‐Met in the presence of AA and UA at pH 7.2. However, the p‐AMTa electrode separates the voltammetric signals of AA, UA and L ‐Met with pronounced oxidation currents. The amperometric current of L ‐Met was increased linearly from 1.0×10^?7 to 1×10^?4 M and the detection limit was found to be 4.12×10^?10 M (S/N=3).     

Quantitative analysis of (styrene/acrylonitrile/methyl methacrylate) co-polymer systems by infrared resonance spectroscopy

A detailed careful analysis of the infrared resonance (IR) spectra of polystyrene (PSt), polymethyl methacrylate (PMMA), polyacrylonitrile (PAN) and their co-mixtures were performed. Through this study the absorption peak area to weight ratios as well as working curves were obtained to test for their reliability as well as their suitability. Satisfactory results were achieved and these working curves were then used to measure the polymerized components of binary and ternary co-polymers. By investigating the acquired data we conclude that the monomer preferential polymeric sequence is St > MMA > AN. A quantitative method to measure P (St/AN/MMA) concentrations by IR spectroscopy is proposed in this work.     

Nonlinear Effects in Asymmetric Catalysis

There is a need for the preparation of enantiomerically pure compounds for various applications. An efficient approach to achieve this goal is asymmetric catalysis. The chiral catalyst is usually prepared from a chiral auxiliary, which itself is derived from a natural product or by resolution of a racemic precursor. The use of non‐enantiopure chiral auxiliaries in asymmetric catalysis seems unattractive to preparative chemists, since the anticipated enantiomeric excess (ee) of the reaction product should be proportional to the ee value of the chiral auxiliary (linearity). In fact, some deviation from linearity may arise. Such nonlinear effects can be rich in mechanistic information and can be synthetically useful (asymmetric amplification). This Review documents the advances made during the last decade in the use of nonlinear effects in the area of organometallic and organic catalysis.     

A Palladium(II)‐Catalyzed Synthesis of Spiroacetals through a One‐Pot Multicomponent Cascade Reaction

Functionalized spiroacetals have been easily prepared in a one‐pot three‐component coupling process that involves the reaction of pentynol derivatives, salicylaldehydes, and amines in the presence of catalytic amounts of a palladium(II) complex (see scheme). Alternatively, oxygen‐substituted spiroacetals can be obtained by using orthoesters as the third component.

     

Three new coordination complexes of cobalt(III), manganese(II), and copper(II) with N,N,O-donor hydrazone ligands: syntheses and structural characterizations

Three new coordination complexes, 2{[Co(L¹)₂]ClO₄} · 0.5CH₃OH (1), [Mn(L²)₂] (2), and [Cu(HL²)(L²)]ClO₄ · 2H₂O (3) have been synthesized from two tridentate N,N,O-donor hydrazone ligands HL¹, 2-acetylpyridine-salicyloylhydrazone, and HL², 2-benzoylpyridine-salicyloylhydrazone, respectively and thoroughly characterized by elemental analysis, FT-IR, UV–Vis, electrochemical, and room temperature magnetic susceptibility measurements. Structures of the complexes have been unequivocally established by single crystal X-ray diffraction technique. Structural analysis reveals that 1 consists of two chemically similar but crystallographically independent cationic [Co(L¹)₂]⁺ units and 2 consists of a neutral [Mn(L²)₂] molecule while 3 consists of a cationic [Cu(HL²)(L²)]⁺ unit. Metal ions display distorted octahedral geometry in 1 and 2 while in 3 it shows a distorted square pyramidal geometry. Ligand conformations around the metal ions are stabilized by the presence of intra-ligand hydrogen bonding in all the complexes. Structure of 3 reveals that a perchlorate ion linked to the complex by hydrogen bonding via a water molecule.     

Hamiltonian cycles in -circulant digraphs

Let D be the circulant digraph with n vertices and connection set {2,3,c}. (Assume D is loopless and has outdegree 3.) Work of S. C. Locke and D. Witte implies that if n is a multiple of 6, c{(n/2)+2,(n/2)+3}, and c is even, then D does not have a hamiltonian cycle. For all other cases, we construct a hamiltonian cycle in D.