Picosecond‐resolved X‐ray absorption spectroscopy at low signal contrast using a hard X‐ray streak camera

A picosecond‐resolving hard‐X‐ray streak camera has been in operation for several years at Sector 7 of the Advanced Photon Source (APS). Several upgrades have been implemented over the past few years to optimize integration into the beamline, reduce the timing jitter, and improve the signal‐to‐noise ratio. These include the development of X‐ray optics for focusing the X‐rays into the sample and the entrance slit of the streak camera, and measures to minimize the amount of laser light needed to generate the deflection‐voltage ramp. For the latter, the photoconductive switch generating the deflection ramp was replaced with microwave power electronics. With these, the streak camera operates routinely at 88 MHz repetition rate, thus making it compatible with all of the APS fill patterns including use of all the X‐rays in the 324‐bunch mode. Sample data are shown to demonstrate the performance.     

Activated Ion ETD Performed in a Modified Collision Cell on a Hybrid QLT-Oribtrap Mass Spectrometer

We describe the implementation and characterization of activated ion electron transfer dissociation (AI-ETD) on a hybrid QLT-Orbitrap mass spectrometer. AI-ETD was performed using a collision cell that was modified to enable ETD reactions, in addition to normal collisional activation. The instrument manifold was modified to enable irradiation of ions along the axis of this modified cell with IR photons from a CO₂ laser. Laser power settings were optimized for both charge (z) and mass to charge (m/z) and the instrument control firmware was updated to allow for automated adjustments to the level of irradiation. This implementation of AI-ETD yielded 1.6-fold more unique identifications than ETD in an nLC-MS/MS analysis of tryptic yeast peptides. Furthermore, we investigated the application of AI-ETD on large scale analysis of phosphopeptides, where laser power aids ETD, but can produce b- and y-type ions because of the phosphoryl moiety’s high IR adsorption. nLC-MS/MS analysis of phosphopeptides derived from human embryonic stem cells using AI-ETD yielded 2.4-fold more unique identifications than ETD alone, demonstrating a promising advance in ETD sequencing of PTM containing peptides.

Synthesis of Enol Esters from Copper(I) Carboxylates Generated from Copper(I) Trifluoromethanesulfonate Benzene Complex

A rapid method for the preparation of copper(I) carboxylates by use of copper(I) trifluoromethanesulfonate benzene complex followed by their conversion to enol esters under mild conditions is presented.     

Formation of smooth, conformal molecular layers on ZnO surfaces via photochemical grafting

We have investigated the photochemical grafting of organic alkenes to atomically flat ZnO(10 ?10) single crystals and ZnO nanorods as a way to produce functional molecule-semiconductor interfaces. Atomic force microscopy shows that photochemical grafting produces highly conformal, smooth molecular layers with no detectable changes in the underlying structure of the ZnO terraces or steps. X-ray photoelectron spectroscopy measurements show that grafting of a methyl ester-terminated alkene terminates near one monolayer, while alkenes bearing a trifluoroacetamide-protected amine form very smooth multilayers. Even with multilayers, it is possible to deprotect the amines and to link a second molecule to the surface with excellent efficiency and without significant loss of molecules from the surface. This demonstrates that the use of photochemical grafting, even in the case of multilayer formation, enables multistep chemical processes to be conducted on the ZnO surface. Photoresponse measurements demonstrate that functionalization of the surface does not affect the ability to induce field effects in the underlying ZnO, thereby suggesting that this approach to functionalization may be useful for applications in sensing and in hybrid organic-inorganic transistors and related devices.     

Bi-functional gold-coated magnetite composites with improved biocompatibility

The effect of gold attachment on the physical characteristics, cellular uptake, gene expression efficiency, and biocompatibility of magnetic iron oxide (MNP) vector was investigated in vitro in BHK21 cells. The surface modification of magnetite with gold was shown to alter the morphology and surface charge of the vector. Nonetheless, despite the differences in the surface charge with and without gold attachment, the surface charge of all vectors were positive when conjugated with PEI/DNA complex, and switched from positive to negative when suspended in cell media containing serum, indicating the adsorption of serum components onto the composite. The cellular uptake of all MNP vectors under the influence of a magnetic field increased when the composite loadings increased, and was higher for the MNP vector that was modified with gold. Both bare magnetite and gold-coated magnetite vectors gave similar optimal gene expression efficiency, however, the gold-coated magnetite vector required a 25-fold higher overall loading to achieve a comparable efficiency as the attachment of gold increased the particle size, thus reducing the surface area for PEI/DNA complex conjugation. The MNP vector without gold showed optimal gene expression efficiency at a specific magnetite loading, however further increases beyond the optimum loading decreased the efficiency of gene expression. The drop in efficiency at high magnetite loadings was attributed to the significant reduction in cellular viability, indicating the bare magnetite became toxic at high intracellular levels. The gene expression efficiency of the gold-modified vector, on the other hand, did not diminish with increasing magnetite loadings. Intracellular examination of both bare magnetite and gold-coated magnetite vectors at 48h post-magnetofection using transmission electron microscopy provided evidence of the localization of both vectors in the cell nucleus for gene expression and elucidated the nuclear uptake mechanism of both vectors. The results of this work demonstrate the efficacy of gold-modified vectors to be used in cellular therapy research that can function both as a magnetically-driven gene delivery vehicle and an intracellular imaging agent with negligible impact on cell viability.     

Synthesis of a designed sesquiterpenoid that forms useful composites with peptides and related oligomers

Efficient desymmetrization of isophthalaldehyde allows a scalable asymmetric synthesis of cinnamylated sesquiterpenoid 1. We have shown that 1 forms useful, property-altered composites with peptides and related oligomers. The current synthesis promises to expand those efforts considerably.     

Metal-free intermolecular oxidative C-N bond formation via tandem C-H and N-H bond functionalization

The development of a novel intermolecular oxidative amination reaction, a synthetic transformation that involves the simultaneous functionalization of both a N-H and C-H bond, is described. The process, which is mediated by an I(III) oxidant and contains no metal catalysts, provides a rapid and green method for synthesizing protected anilines from simple arenes and phthalimide. Mechanistic investigations indicate that the reaction proceeds via nucleophilic attack of the phthalimide on an aromatic radical cation, as opposed to the electrophilic aromatic amination that has been reported for other I(III) amination reactions. The application of this new reaction to the synthesis of a variety of substituted aniline derivatives is demonstrated.     

Characterization and Diagnostic Value of Amino Acid Side Chain Neutral Losses Following Electron-Transfer Dissociation

Using a large set of high mass accuracy and resolution ETD tandem mass spectra, we characterized ETD-induced neutral losses. From these data we deduced the chemical formula for 20 of these losses. Many of them have been previously observed in electron-capture dissociation (ECD) spectra, such as losses of the side chains of arginine, aspartic acid, glutamic acid, glutamine, asparagine, leucine, histidine, and carbamidomethylated cysteine residues. With this information, we examined the diagnostic value of these amino acid-specific losses. Among 1285 peptide–spectrum matches, 92.5% have agreement between neutral loss-derived peptide amino acid composition and the peptide sequences. Moreover, we show that peptides can be uniquely identified by using only the accurate precursor mass and amino acid composition based on neutral losses; the median number of sequence candidates from an accurate mass query is reduced from 21 to 8 by adding side chain loss information. Besides increasing confidence in peptide identification, our findings suggest the potential use of these diagnostic losses in ETD spectra to improve false discovery rate estimation and to enhance the performance of scoring functions in database search algorithms.     

Across-site patterns of modulation detection in listeners with cochlear implants

In modern cochlear implants, much of the information required for recognition of important sounds is conveyed by temporal modulation of the charge per phase in interleaved trains of electrical pulses. In this study, modulation detection thresholds (MDTs) were used to assess listeners' abilities to detect sinusoidal modulation of charge per phase at each available stimulation site in their 22-electrode implants. Fourteen subjects were tested. MDTs were found to be highly variable across stimulation sites in most listeners. The across-site patterns of MDTs differed considerably from subject to subject. The subject-specific patterns of across-site variability of MDTs suggest that peripheral site-specific characteristics, such as electrode placement and the number and condition of surviving neurons, play a primary role in determining modulation sensitivity. Across-site patterns of detection thresholds (T levels), maximum comfortable loudness levels (C levels) and dynamic ranges (DRs) were not consistently correlated with across-site patterns of MDTs within subjects, indicating that the mechanisms underlying across-site variation in these measures differed from those underlying across-site variation in MDTs. MDTs sampled from multiple sites in a listener's electrode array might be useful for diagnosing across-subject differences in speech recognition with cochlear implants and for guiding strategies to improve the individual's perception.     

A computer simulation study of imaging flexural inhomogeneities using plate-wave diffraction tomography

This paper investigates the feasibility of plate-wave diffraction tomography for the reconstruction of flexural inhomogeneities in plates using the results of computer simulation studies. The numerical implementation of the fundamental reconstruction algorithm, which has recently been developed by Wang and Rose [C.H. Wang, L.R.F. Rose, Plate-wave diffraction tomography for structural health monitoring, Rev. Quant. Nondestr. Eval. 22 (2003) 1615-1622] is investigated addressing the essential effects of applying the discrete form of the Fourier diffraction theorem for solving the inverse problem as discussed by Kak and Slaney [A.C. Kak, M. Slaney, Principles of Computerized Tomographic Imaging, IEEE Press, New York, 1988] for the acoustic case, viz. diffraction limited sensitivity, influence of weak scatterer assumption, damage location and scatter field data processing in time and Fourier space as well as experimental limitations such as finite receiver length and limited views. The feasibility of the imaging technique is investigated for cylindrical inhomogeneities of various severities and relative position within the interrogation space and a normal incident interrogation configuration. The results show that plate-wave diffraction tomography enables the quantitative reconstruction of location, size and severity of plate damage with excellent sensitivity and offers the potential for detecting corrosion thinning, disbonds and delamination damage in structural integrity management applications.