Cigarette smoke extract induced protein phosphorylation changes in human microvascular endothelial cells in vitro

Phosphorylation is the most widely studied posttranslational modification (PTM) and is an important regulatory mechanism used during cellular responses to external stimuli. The kinases and phosphatases that regulate protein phosphorylation are known to be affected in many human diseases. Cigarette smoking causes cardiovascular disease (CVD). Endothelial cells play a pivotal role in CVD initiation and development; however, there have been limited investigations of the specific signaling cascades and protein phosphorylations activated by cigarette smoke in endothelial cells. The purpose of this research was to better understand the differential protein phosphorylation in endothelial cells stimulated with extracts of cigarette smoke total particulate matter (CS-TPM) in vitro. Human microvascular endothelial cells were exposed in vitro to CS-TPM at concentrations that were shown to cause endothelial cell dysfunction. The phosphorylated proteins were isolated using phosphoprotein-specific chromatography, followed by enzymatic digestion and nano-flow capillary liquid chromatography (ncap-LC) coupled to high resolution mass spectrometry. This study putatively identified 94 proteins in human microvascular endothelial cells that were differentially bound to a phosphoprotein-specific chromatography column following exposure to CS-TPM suggesting differential phosphorylation. Pathway analysis has also been conducted and confirmations of several observations have been made using immunoaffinity-based techniques (e.g., Western blotting). Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Stable and Unstable Solitary-Wave Solutions of the Generalized Regularized Long-Wave Equation

Summary. Investigated here are interesting aspects of the solitary-wave solutions of the generalized Regularized Long-Wave equation For p>5 , the equation has both stable and unstable solitary-wave solutions, according to the theory of Souganidis and Strauss. Using a high-order accurate numerical scheme for the approximation of solutions of the equation, the dynamics of suitably perturbed solitary waves are examined. Among other conclusions, we find that unstable solitary waves may evolve into several, stable solitary waves and that positive initial data need not feature solitary waves at all in its long-time asymptotics. Received March 28, 2000; accepted August 24, 2000 %%Online publication November 15, 2000 Communicated by Thanasis Fokas     

Operator algebras and a theorem of Dieudonne

LetA be aC*-algebra with second dualA″. Let (φ _n)(n=1,...) be a sequence in the dual ofA such that limφ _n(a) exists for eacha εA. In general, this does not imply that limφ _n(x) exists for eachx εA″. But if limφ _n(p) exists whenever p is the range projection of a positive self-adjoint element of the unit ball ofA, then it is shown that limφ _n(x) does exist for eachx inA″. This is a non-commutative generalisation of a celebrated theorem of Dieudonné. A new proof of Dieudonné’s theorem, for positive measures, is given here. The proof of the main result makes use of Dieudonné’s original theorem.     

A screening assay for the tetrachlorodibenzo-p-dioxin receptor using the [125I]iodovaleramide derivative of trichlorodibenzo-p-dioxin as the binding ligand

A relatively simple assay method for the putative cytosolic 'receptor' that binds 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and related compounds is described. The assay is based on specific binding of [125I]dioxin to cytosol 'receptor' protein. Saturation is ensured by competition experiments in which unlabeled TCDD and other competitors displace the radiolabeled ligand from specific binding sites. This assay has been applied to estimation of levels of 'receptor' in cytosol.     

CIRCE: a dedicated storage ring for coherent THz synchrotron radiation

We present the concepts for an electron storage ring dedicated to and optimized for the production of stable coherent synchrotron radiation (CSR) over the far-infrared terahertz wavelength range from 200 μm to about 1 cm. CIRCE (Coherent InfraRed CEnter) will be a 66 m circumference ring located on top of the ALS booster synchrotron shielding tunnel and using the existing ALS injector. This location provides enough floor space for both the CIRCE ring, its required shielding, and numerous beamlines. We briefly outline a model for CSR emission in which a static bunch distortion induced by the synchrotron radiation field is used to significantly extend the stable CSR emission towards higher frequencies. This model has been verified with experimental CSR results. We present the calculated CIRCE photon flux where a gain of 6–9 orders of magnitude is shown compared to existing far-IR sources. Additionally, the particular design of the dipole vacuum chamber has been optimized to allow an excellent transmission of these far-infrared wavelengths. We believe that the CIRCE source can be constructed for a modest cost.     

Noise reduction efforts for the ALS infrared beamlines

The quality of infrared microscopy and spectroscopy data collected at synchrotron based sources is strongly dependent on signal-to-noise. We have successfully identified and suppressed several noise sources affecting beamlines 1.4.2, 1.4.3, and 1.4.4 at the advanced light source (ALS), resulting in a significant increase in the quality of FTIR spectra obtained. In this paper, we present our methods of noise source analysis, the negative effect of noise on the infrared beam quality, and the techniques used to reduce the noise. These include reducing the phase noise in the storage ring radio-frequency (RF) system, installing an active mirror feedback system, analyzing and changing physical mounts to better isolate portions of the beamline optics from low-frequency environmental noise, and modifying the input signals to the main ALS RF system. We also discuss the relationship between electron beam energy oscillations at a point of dispersion and infrared beamline noise.     

The detection of piroxicam, tenoxicam and their metabolites in equine urine by electrospray ionisation ion trap mass spectrometry

An investigation has been conducted into the metabolism and urinary excretion of orally administered piroxicam and tenoxicam in the horse. The major component detected in urine after the administration of piroxicam was 5'-hydroxypiroxicam, which was detectable up to 24 h post-administration. Unchanged piroxicam was present only as a minor component. In contrast, unchanged tenoxicam was the major component observed after the administration of tenoxicam, being detectable for 72 h post-administration, while 5'-hydroxytenoxicam was a minor component. Phase II beta-glucuronide conjugation in each case was found to be negligible. The ion trap mass spectral characteristics of piroxicam, tenoxicam, 5'-hydroxypiroxicam and 5'-hydroxytenoxicam under electrospray ionisation conditions were examined in some detail.     

Molecular orbital studies of the NMR hydrogen bond shift

Magnetic shielding constants are calculated for the protons in XOH and XOH…OH₂ (XH, CH₃, NH₂, OH and F) molecules using a slightly extended set of atomic functions modified by gauge factors. These results are used to determine theoretical values for the NMR hydrogen bond shifts in the XOH…OH₂ systems. Such theoretical data are consistent with the few available experimental data. An analysis of the theoretical results reveals that there are three major types of shielding contribution to the NMR hydrogen bond shift; (a) a deshielding change due to the variation of the local currents on the hydrogen bonded proton; (b) a reduction in shielding from currents localized on the oxygen atom of the proton donor; (c) a deshielding contribution from currents induced on the oxygen atom of the proton acceptor. Except for the water dimer, contributions (a), (b) and (c) are of comparable importance for changes in isotropic shielding. For (H₂O)₂ contributions (a) and (c) are somewhat more important than contribution (b). Contribution (c) is almost totally responsible for the changes in the anistropies of the shielding tensors associated with the hydrogen bonded protons. The proton shielding anisotropy changes which occur on hydrogen bond formation are generally much larger than the corresponding variations in the isotropic values of the shielding tensors. This suggests that proton magnetic shielding anisotropies may be more sensitive measures of features of hydrogen bonding than are isotropic proton shielding constants.