Mutants of Ca2+‐regulated Photoprotein Obelin for Site‐specific Conjugation

Color variants of Ca²⁺‐regulated photoprotein obelin were shown to be an important tool for dual‐analyte binding assay. To provide site‐directed conjugation with biospecific molecules, several obelin color mutants carrying unique cysteine residues were obtained and characterized for their novel properties. A pair of obelins Y138F,A5C and W92F,H22E,D12C was found to be most suitable (in terms of high bioluminescent activity and stability) as reporters in simultaneous assay of two targets in a sample. Availability of SH‐groups, accessible for chemical modification, essentially simplifies the synthesis of biospecific conjugates, increases their yield and conserves obelins' bioluminescence activity. Conjugates with immunoglobulin and oligonucleotide were produced and successfully applied in single nucleotide polymorphism genotyping.     

Electroless Deposition of III–V Semiconductor Nanostructures from Ionic Liquids at Room Temperature

Group III–V semiconductor nanostructures are important materials in optoelectronic devices and are being researched in energy‐related fields. A simple approach for the synthesis of these semiconductors with well‐defined nanostructures is desired. Electroless deposition (galvanic displacement) is a fast and versatile technique for deposition of one material on another and depends on the redox potentials of the two materials. Herein we show that GaSb can be directly synthesized at room temperature by galvanic displacement of SbCl₃/ionic liquid on electrodeposited Ga, on Ga nanowires, and also on commercial Ga. In situ AFM revealed the galvanic displacement process of Sb on Ga and showed that the displacement process continues even after the formation of GaSb. The bandgap of the deposited GaSb was 0.9±0.1 eV compared to its usual bandgap of 0.7 eV. By changing the cation in the ionic liquid, the redox process could be varied leading to GaSb with different optical properties.     

Influence of Synthesis pH and Oxidative Strength of the Catalyzing Acid on the Morphology and Chemical Structure of Hydrothermal Carbon

A specific control of the morphology and chemical structure of hydrothermal carbon (HTC) is of crucial importance for its application, both in catalyst supports or electrochemical devices. Here we show how the morphology, that is, particles size and homogeneity, and the distribution of functional groups can be controlled by the control of the synthesis pH of the hydrothermal carbonization. A complementary analysis of liquid byproducts by HPLC provides useful information on the nature of the polymeric species produced during the poly‐condensation in the hydrothermal process and reveals the potential implementation of the process into the biorefinery concept. The acidic byproducts levulinic acid and formic acid determine the hydrothermal carbonization autocatalytically by additional supply of protons to the reaction medium. Thus, for a starting pH>3, only minor structural differences can be detected for HTC. The use of oxidizing acids favors higher yields of HTC and improves carbonization towards higher condensed carbon domains. Scaling up the process in a stirred 2 L batch reactor favors carbonization leading to higher condensed carbonaceous products. The relative trends of pH variation are maintained.     

Optical bistability in a nonlinear optical coupler with a negative index channel

We discuss a novel kind of nonlinear coupler with one channel filled with a negative index metamaterial. The opposite directionality of the phase velocity and the energy flow in the negative index metamaterial channel facilitates an effective feedback mechanism that leads to optical bistability and gap soliton formation.     

Comparison of two derivatizing agents for the simultaneous determination of selenite and organoselenium species by gas chromatography and atomic emission detection after preconcentration using solid-phase microextraction

Two methods for the simultaneous determination of selenite and two organoselenium compounds, dimethylselenide (DMSe) and dimethyldiselenide (DMDSe), are proposed. Both methods involve sample preconcentration by solid-phase microextraction (SPME) and capillary gas chromatography coupled to atomic emission detection (GC-AED). The main difference between the methods is the derivatizing agent used to complex the inorganic species: sodium tetraethylborate and 4,5-dichloro-1,2-phenylenediamine. The parameters affecting the derivatization and preconcentration steps, chromatographic separation as well as detection of the compounds were optimized. Direct immersion (DI) mode and a relatively long extraction time were selected for the method involving the formation of the piazselenol complex, better sensitivity being achieved for the three analytes under study. In this case, detection limits ranged between 3 and 25 ng L(-1), depending on the compound. Headspace mode (HS) and extraction times of 20 min were selected for the method involving tetraalkylborate, and detection limits of between 7.3 and 55 ng L(-1) were obtained. DMSe and Se(IV) were found in several of the water samples analyzed at concentrations of 0.07-1.0 ng mL(-1).     

IR laser and heat-induced changes in annulus fibrosus collagen structure

The purpose of this study was to characterize essential changes in the structure of annulus fibrosus (AF) after hydrothermal and infrared (IR) laser treatment and to correlate these results with alterations in tissue state. Polarization-sensitive optical coherence tomography imaging was used to measure collagen birefringence in AF. Differential scanning calorimetry was used as a complementary technique, providing detailed information on thermodynamic processes in the tissue. Birefringence, peak of the denaturation endotherm, and the enthalpy of denaturation (DeltaHm) were determined before and after hydrothermal heat treatment (85 degrees C for 15 min) and non-ablative Er:glass fiber laser exposures on AF in the whole disk (vertebrae-disk-vertebrae complex). Our data have demonstrated quantitative differences between results of laser and hydrothermal heating. Birefringence did not disappear and DeltaHm did not change after treatment in the water bath, but loss of birefringence and a decrease in the enthalpy did occur after laser exposure. These results could be explained by the photomechanical effect of laser irradiation. We suggest that thermo-mechanical stress played a dominant role in the disruption of the collagen network of AF under non-homogeneous laser heating.     

Palladium-catalyzed asymmetric phosphination. Scope, mechanism, and origin of enantioselectivity

Asymmetric cross-coupling of aryl iodides (ArI) with secondary arylphosphines (PHMe(Ar'), Ar' = (2,4,6)-R3C6H2; R = i-Pr (Is), Me (Mes), Ph (Phes)) in the presence of the base NaOSiMe3 and a chiral Pd catalyst precursor, such as Pd((R,R)-Me-Duphos)(trans-stilbene), gave the tertiary phosphines PMe(Ar')(Ar) in enantioenriched form. Sterically demanding secondary phosphine substituents (Ar') and aryl iodides with electron-donating para substituents resulted in the highest enantiomeric excess, up to 88%. Phosphination of ortho-substituted aryl iodides required a Pd(Et-FerroTANE) catalyst but gave low enantioselectivity. Observations during catalysis and stoichiometric studies of the individual steps suggested a mechanism for the cross-coupling of PhI and PHMe(Is) (1) initiated by oxidative addition to Pd(0) yielding Pd((R,R)-Me-Duphos)(Ph)(I) (3). Reversible displacement of iodide by PHMe(Is) gave the cation [Pd((R,R)-Me-Duphos)(Ph)(PHMe(Is))][I] (4), which was isolated as the triflate salt and crystallographically characterized. Deprotonation of 4-OTf with NaOSiMe3 gave the phosphido complex Pd((R,R)-Me-Duphos)(Ph)(PMeIs) (5); an equilibrium between its diastereomers was observed by low-temperature NMR spectroscopy. Reductive elimination of 5 yielded different products depending on the conditions. In the absence of a trap, the unstable three-coordinate phosphine complex Pd((R,R)-Me-Duphos)(PMeIs(Ph)) (6) was formed. Decomposition of 5 in the presence of PhI gave PMeIs(Ph) (2) and regenerated 3, while trapping with phosphine 1 during catalysis gave Pd((R,R)-Me-Duphos)(PHMe(Is))2 (7), which reacted with PhI to give 3. Deprotonation of 1:1 or 1.4:1 mixtures of cations 4-OTf gave the same 6:1 ratio of enantiomers of PMeIs(Ph) (2), suggesting that the rate of P inversion in 5 was greater than or equal to the rate of reductive elimination. Kinetic studies of the first-order reductive elimination of 5 were consistent with a Curtin-Hammett-Winstein-Holness (CHWH) scheme, in which pyramidal inversion at the phosphido ligand was much faster than P-C bond formation. The absolute configuration of the phosphine (SP)-PMeIs(p-MeOC6H4) was determined crystallographically; NMR studies and comparison to the stable complex 5-Pt were consistent with an RP-phosphido ligand in the major diastereomer of the intermediate Pd((R,R)-Me-Duphos)(Ph)(PMeIs) (5). Therefore, the favored enantiomer of phosphine 2 appeared to be formed from the major diastereomer of phosphido intermediate 5, although the minor intermediate diastereomer underwent P-C bond formation about three times more rapidly. The effects of the diphosphine ligand, the phosphido substituents, and the aryl group on the ratio of diastereomers of the phosphido intermediates Pd(diphos*)(Ar)(PMeAr'), their rates of reductive elimination, and the formation of three-coordinate complexes were probed by low-temperature 31P NMR spectroscopy; the results were also consistent with the CHWH scheme.     

Sample handling strategies for the determination of persistent trace organic contaminants from biota samples

Even after emergence of most advanced instrumental techniques for the final separation, detection, identification and determination of analytes, sample handling continues to play a basic role in environmental analysis of complex matrices. In fact, sample preparation steps are often the bottleneck for combined time and efficiency in many overall analytical procedures. Thus, it is not surprising that, in the last two decades, a lot of effort has been devoted to the development of faster, safer, and more environment friendly techniques for sample extraction and extract clean up, prior to actual instrumental analysis. This article focuses on the state of the art in sample preparation of environmental solid biological samples dedicated to persistent organic pollutants (POPs) analysis. Extraction techniques such as Soxhlet extraction, sonication-assisted extraction, supercritical fluid extraction (SFE), microwave-assisted extraction (MAE), pressurised liquid extraction (PLE) and matrix solid-phase dispersion (MSPD) are reviewed and their most recent applications to the determination of POPs in biota samples are provided. Additionally, classical as well as promising novel extraction/clean-up techniques such as solid phase microextraction (SPME) are also summarized. Finally, emerging trends in sample preparation able to integrate analytes extraction and their adequate clean-up are presented.     

Click multivalent neoglycoconjugates as synthetic activators in cell adhesion and stimulation of monocyte/machrophage cell lines

The efficient synthesis of fluorescent and non-fluorescent multivalent neoglycoconjugates is described by means of the Cu(i) catalyzed azide-alkyne 1,3-dipolar cycloaddition ("click-chemistry"). A well-defined glycopolymer, glycocyclodextrin or glycocluster architecture displaying galactose or lactose epitopes has been chosen. Cellular assays using U-937 and RAW 264.7 monocyte/macrophage cells showed that these glycocompounds have the capability to act as synthetic activators mimicking the lipopolysaccharide (LPS) effects. Thus, the click compounds promote cell adhesion and stimulation of monocytes, measured as an increase in the amount of TNFalpha, facilitating their differentiation to macrophages.     

Interaction of unmodified and partially silylated nanosilica with red blood cells

Interaction of red blood cells (RBCs) with unmodified and partially (50%) silylated fumed silica A-300 (nanosilica)was studied by microscopic, XRD and thermally stimulated depolarisation current (TSDC) methods. Nanosilica at a low concentration C _A-300 < 0.01 wt.% in buffered aqueous suspension is characterised by a weak haemolytic effect on RBCs. However, at C _A-300 = 1 wt% all RBCs transform into shadow corpuscles because of 100% haemolysis. Partial (one-half) hydrophobization of nanosilica leads to reduction of the haemolytic effect in comparison with unmodified silica at the same concentrations. A certain portion of the TSDC spectra of the buffered suspensions with RBC/A-300 is independent of the amounts of silica. However, significant portions of the low-and high-temperature TSDC bands have a lower intensity at C _A-300 = 1 wt% than that for RBCs alone or RBC/A-300 at C _A-300 = 0.01 wt.% because of structural changes in RBCs. Results of microscopic and XRD investigations and calculations using the TSDC-and NMR-cryoporometry suggest that the intracellular structures in RBCs (both organic and aqueous components) depend on nanosilica concentration in the suspension.