Aptamer-based enantioselective competitive binding assay for the trace enantiomer detection

The development of highly enantioselective assays and sensors has received much attention for the determination of enantiomeric impurities at a low level. For chiral compounds, the efficient monitoring of the in selection procedure has allowed the isolation of nucleic acid aptamers which are able to strongly discriminate the target enantiomers. In this paper, we demonstrated for the first time that an aptamer can be successfully used to design a highly enantioselective tool for the trace enantiomer detection. The aptamer-based stereoselective assay was developed using an affinity capillary electrophoresis-based competitive, homogeneous format and an on-capillary mixing approach. Detection of as low as 0.01% of the minor enantiomer in a nonracemic mixture can be achieved, in a short analysis time (<5 min).     

Practical constraints in the kinetic plot representation of chromatographic performance data: theory and application to experimental data

It is demonstrated that the kinetic plot representation of experimental plate height data can also account for practical constraints on the column length, the peak width, the viscous heating, and the mobile-phase velocity without needing any iterative solution routine. This implies that the best possible kinetic performance to be expected from a given tested support under any possible set of practical optimization constraints can always be found using a directly responding calculation spreadsheet template. To show how the resulting constrained kinetic plots can be used as a powerful design and selection tool, the method has been applied to a series of plate height measurements performed on a number of different commercial columns for the same component (butyl-parabene) and mobile-phase composition. The method, for example, allows one to account for the fact that the advantageous solutions displayed by the silica monolith and 5 microm particle columns in the large plate number range of the free kinetic plot are no longer accessible if applying a maximal column length constraint of Lmax = 30 cm. In the plate number range that remains accessible, the investigated sub-2 mum particle columns in any case perform (at least for the presently considered parabene separation) better than the 3.5 mum particle columns or silica monolith, especially if considering the use of system pressures exceeding 400 bar. The constrained kinetic plot method can also be used to select the best-suited column length from an available product gamma to perform a separation with a preset number of plates. One of the optimization results that is obtained in this case is that sometimes a significant gain in analysis time can be obtained by selecting a longer column, yielding the desired plate number at a larger velocity than that for a shorter column.     

Conjugation of keto fatty acids to glutathione in plant tissues. Characterization and quantification by HPLC-tandem mass spectrometry

Both biotic and abiotic stress activate the oxylipin pathway in plants. As reactive electrophile species (RES), some oxylipins are expected to bind cellular nucleophiles in a Micha?l-type addition reaction. Using the HPLC-tandem mass spectrometry techniques, we have established the analytical basis for the investigation of oxylipin conjugation to glutathione (GSH) in plant extracts. The GSH adducts to the four keto fatty acid isomers issued from both linoleic and linolenic acids were first produced and their mass spectrometric features analyzed in the positive electrospray ionization mode. In all cases, the main fragmentation (MS2 mode) of the pseudomolecular ion leads to the neutral loss of a glutamyl moiety (-129 Da), affording an ion that gives structural information upon an additional fragmentation (MS3 mode). The glutamyl loss was confirmed by the analysis of other GSH adducts to oxylipin RES and appeared as being characteristic of GSH adducts. It is thus proposed to search GSH adducts in plant extracts by HPLC-MS/MS, using initially the neutral loss mode and then the MS2 mode to further characterize the identified compounds. This methodology was successfully applied to the analysis of GSH adducts upon infiltration into leaves of the four previous keto fatty acids at 5 mM, a concentration inducing cell death. The production of GSH adducts to oxylipin RES was observed for the first time in plant tissues. Furthermore, the levels of adduct production explain in part the observed GSH depletion. These results support the role of RES in altering protein activities and cellular redox balance of plant cells, via addition reactions to cellular nucleophiles.     

Propagation losses and gain measurements in erbium-doped fluoride glass channel waveguides by use of a double-pass technique

We have studied Er3+, Yb3+, and Ce3+ codoped microchannel waveguides that were developed by two methods: ionic exchange for heavy metal fluoride glasses [ZrF4-BaF2-AlF3-CeF3 (ZBAC)] and vapor phase deposition for transition metal fluoride glasses [PbF2-ZnF2-GaF3 (PZG)] by using a double-pass technique. For the first time to our knowledge, the measurement of propagation losses and amplification tests were carried out by use of the same experimental setup, leading to complete characterization of the waveguides. Net gains higher than 1 dB/cm were achieved in ZBAC Er/Ce single-mode fluoride glass waveguides.     

Multiplexed holographic transmission gratings recorded in holographic polymer-dispersed liquid crystals: static and dynamic studies

The optimization of the experimental parameters of two multiplexed holographic transmission gratings recorded in holographic polymer-dispersed liquid crystals is investigated. Two methods are used to record the holograms: simultaneous and sequential multiplexing. These two processes are optimized to produce two multiplexed Bragg gratings that have the same and the highest possible diffraction efficiencies in the first order. The two methods show similar results when suitable recording parameters are used. The parameters of the recorded gratings (mainly the refractive-index modulation) are retrieved by use of an extension of the rigorous coupled-wave theory to multiplexed gratings. Finally, the response of the holograms to an electric field is studied. We demonstrate few coupling effects between the behavior of both gratings, and we expect a possibility of switching from one grating to the other.     

On the effects of reflected waves in transient shear wave elastography

In recent years, novel quantitative techniques have been developed to provide noninvasive and quantitative stiffness images based on shear wave propagation. Using radiation force and ultrafast ultrasound imaging, the supersonic shear imaging technique allows one to remotely generate and follow a transient plane shear wave propagating in vivo in real time. The tissue shear modulus, i.e., its stiffness, can then be estimated from the shear wave local velocity. However, because the local shear wave velocity is estimated using a time-of- flight approach, reflected shear waves can cause artifacts in the estimated shear velocity because the incident and reflected waves propagate in opposite directions. Such effects have been reported in the literature as a potential drawback of elastography techniques based on shear wave speed, particularly in the case of high stiffness contrasts, such as in atherosclerotic plaque or stiff lesions. In this letter, we present our implementation of a simple directional filter, previously used for magnetic resonance elastography, which separates the forward- and backward-propagating waves to solve this problem. Such a directional filter could be applied to many elastography techniques based on the local estimation of shear wave speed propagation, such as acoustic radiation force imaging (ARFI), shearwave dispersion ultrasound vibrometry (SDUV), needle-based elastography, harmonic motion imaging, or crawling waves when the local propagation direction is known and high-resolution spatial and temporal data are acquired.     

Chalcogenide coatings of Ge15Sb20S65 and Te20As30Se50

Chalcogenide coatings are investigated to obtain either optical components for spectral applications or optochemical sensors in the mid-infrared. The deposition of Ge(15)Sb(20)S(65) and Te(20)As(30)Se(50) chalcogenide glasses is performed by two physical techniques: electron-beam and pulsed-laser deposition. The quality of the film is analyzed by scanning electron microscopy, atomic force microscopy, and energy dispersive spectroscopy to characterize the morphology, topography, and chemical composition. The optical properties and optical constants are also determined. A CF(4) dry etching is performed on these films to obtain a channeled optical waveguide. For a passband filter made by electron-beam deposition, cryolite as a low-refractive-index material and chalcogenide glasses as high-refractive-index materials are used to favor a large refractive-index contrast. A shift of a centered wavelength of a photosensitive passband filter is controlled by illumination time.