A comparative investigation into the effect of chronic alcohol feeding on the myocardium of normotensive and hypertensive rats: an electrophoretic and biochemical study

We investigated whether the imposition of chronic alcohol in hypertension leads to greater biochemical and cellular abnormalities of the myocardium than those arising in normotension. Fifteen-week-old spontaneously hypertensive rats (SHR) and Wistar Kyoto (WKY) rats were fed ethanol-containing diets for six weeks. Particular attention was focused on the composition of contractile proteins identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), fractional rate of protein synthesis, and synthesis rates relative to RNA (RNA activity) or DNA (cellular efficiency). In addition, myocardial enzymes and adenine nucleotides were measured. In both SHR and WKY rats chronic ethanol caused a general decrease in the contents of all nine contractile proteins with myosin heavy chain predominantly affected. Fractional rates of mixed (i.e., total) and myofibrillary proteins remained unaltered in both WKY rats and SHR, as were cellular efficiencies. The RNA activity was significantly reduced in ethanol-treated SHR but not in WKY rats. In ethanol-treated SHR, cardiac creatine kinase (CK) and malate dehydrogenase (MDH) activities were increased, AMP levels were elevated, whilst ATP levels and the energy charge were reduced. In WKY rats, the only significant change related to increased aspartate aminotransferase activities in response to alcohol feeding. Although there were only subtle differences between the response of the normotensive and hypertensive rats due to ethanol dosage, the reduced ATP levels and increased CK and MDH activities in SHR may reflect a greater susceptibility to ischaemic damage. Reduced contractile protein content, particularly myosin heavy chain, may contribute to contractile defects, a common feature of subclinical and clinical alcoholic cardiomyopathy.     

Spectroscopic Determination of Cholinesterase Activity and Inhibition in Sol-Gel Media

Biological activity of cholinesterases can be determined by optically monitoring the enzymatic reaction with indophenyl acetate, (N-4′-acetoxyphenyl)-4-quinone imine. At pH 8.0 cholinesterases hydrolyze this yellow dye to yield a blue reaction product. Cholinesterase inhibitors reduce the rate of this hydrolysis. Thus, by monitoring absorbance of the hydrolysis product at its maximum (630 nm) as a function of time, reaction rates of both cholinesterase activity and cholinesterase inhibition may be quantified spectroscopically. Using this technique, we measured the enzymatic activity of butyrylcholinesterase (BuChE) molecules encapsulated in tetramethyl orthosilicate (TMOS) silicate gel-glass prepared by hydrolysis and condensation. This activity is reduced, in a concentration-dependent manner, by the reversible cholinesterase inhibitors 1,5-bis(4-allyldimethyl-ammoniumphenyl) pentan 3-one dibromide (BADAPP) and 9-amino-1,2,3,4-tetrahydroacridine (THA; tacrine, Cognex). The gel-glasses are rigid, and compact, transparent and porous enough to allow reagents to diffuse in and out.     

The compositional structure of highly turbulent piloted premixed flames issuing into a hot coflow

Simultaneous line measurements of major species and temperature by the Raman–Rayleigh technique, combined with CO two-photon laser-induced fluorescence and crossed-plane OH planar laser-induced fluorescence have been applied to a series of flames in the Piloted Premixed Jet Burner (PPJB). The PPJB is capable of stabilizing highly turbulent premixed jet flames through the use of a stoichiometric pilot and a large coflow of hot combustion products. Four flames with increasing jet velocities and constant jet equivalence ratios are examined in this paper. The characteristics of these four flames range from stable flame brushes with reaction zones that can be described as thin and “flamelet-like” to flames that have thickened reaction zones and exhibit extinction re-ignition behaviour. Radial profiles of the mean temperature are reported, indicating the mean thermal extent of the pilot and spatial location of the mean flame brush. Measurements of carbon monoxide (CO) and the hydroxyl radical (OH) reveal a gradual decrease in the conditional mean as the jet velocity is increased and the flame approaches extinction. Experimental results for the conditional mean temperature gradient show a progressive trend of reaction zone thickening with increasing jet velocities, indicating the increased interaction of turbulence with the reaction zone at higher turbulence levels. For the compositions examined, the product of CO and OH mole fractions ([CO][OH]) is shown to be a good qualitative indicator for the net rate of production of carbon dioxide. The axial variation of [CO][OH] is shown to correlate well with the mean chemi-luminescence of the flames including the extinction re-ignition regions. The experimental findings reported in this paper further support the hypothesis of an initial ignition region followed by extinction and re-ignition regions for certain PPJB flames.     

UHV microscopy of the reconstructed Au(001) surface

We investigate the microstructure of the reconstructed Au(001) surface using ultra-high vacuum transmission electron microscopy (UHV-TEM). Bulk single crystal Au(001) surfaces were prepared via standard metallographic techniques followed by repetitive cleaning of the surface with ion milling and annealing. After a clean surface was obtained, the (001) surface was found to reconstruct into two nearly orthogonal domains of dimensions (5 × n) where n ranges between 15 and 21. The unit cell vectors of the surface cell are parallel to the 110 directions of the unreconstructed fcc (001) surface. Analysis of the diffuse scattering and dark field micrographs indicates that the surface is sheared with a complicated domain and periodicity structure which depends upon the local geometry of the substrate.     

Synthesis and evaluation of sphinganine analogues of KRN7000 and OCH

[structures: see text] The phytosphingosine-containing alpha-galactosylceramides (alpha-GalCers), KRN7000 and OCH, have been shown to activate NKT cells via interaction with CD1d, a member of the CD1 family of antigen presenting proteins. Evidence from KRN7000 stimulation of NKT cells suggests that alpha-GalCers may have applications in the treatment or prevention of a range of viral, bacterial, and autoimmune conditions. Moreover, OCH, a truncated analogue of KRN7000, appears to induce a T(H)2 bias, which could have implications for the treatment of autoimmune and inflammatory conditions. We have prepared the direct sphinganine-containing analogues of KRN7000 and OCH, 1 and 2, and found them to be comparable in activity to the parent compounds in inducing the release of IL-2, IL-4, and IFNgamma. In addition, compound 2 leads to a cytokine bias similar to that seen with OCH. This is significant because sphinganines are more easily accessed than phytosphingosines, which should facilitate SAR studies.     

Tilted superstructure fiber grating used as a Fourier-transform spectrometer

We propose and demonstrate a new method for fabricating a compact all-fiber spectrometer by using a tilted superstructure fiber grating to couple light out of a fiber core and produce an interference pattern in the near field. The Fourier transform of the interference pattern serves as a direct measurement of the optical input spectrum, and the superstructure grating design offers several degrees of freedom that permit control over the resolution and bandwidth of the spectrometer. For single-wavelength operation, the proposed scheme offers the possibility of making wavelength measurements with picometer-level precision over a broad 80-nm wavelength range while simultaneously providing coarser precision over a 160-nm range.     

Recent developments in X-UV optics and X-UV diagnostics

Metrology of XUV beams (X-ray lasers, high-harmonic generation and VUV free-electron lasers) is of crucial importance for the development of applications. We have thus developed several new optical systems enabling us to measure the optical properties of XUV beams. By use of a Michelson interferometer working as a Fourier-transform spectrometer, the line shapes of different X-ray lasers have been measured with a very high accuracy (/10^-6). Achievement of the first XUV wavefront sensor has enabled us to measure the beam quality of laser-pumped as well as discharge-pumped X-ray lasers. A capillary discharge X-ray laser has demonstrated a very good wavefront allowing us to achieve an intensity as high as 3×10¹⁴ Wcm^-2 by focusing with a f=5 cm mirror. The sensor accuracy has been measured using a calibrated spherical wave generated by diffraction. The accuracy has been estimated to be as good as /120 at 13 nm. Commercial developments are underway. At Laboratoire dOptique Appliquée, we are setting up a new beamline based on high-harmonic generation in order to start the femtosecond, coherent XUV optic . PACS 07.85.Nc;32.70.Jz;41.50.+h;42.15.Dp;42.55.Vc;52.70.La     

Picosecond photoreduction of inducible nitric oxide synthase by rhenium(I)-diimine wires

In a continuing effort to unravel mechanistic questions associated with metalloenzymes, we are developing methods for rapid delivery of electrons to deeply buried active sites. Herein, we report picosecond reduction of the heme active site of inducible nitric oxide synthase bound to a series of rhenium-diimine electron-tunneling wires, [Re(CO)3LL']+, where L is 4,7-dimethylphenanthroline and L' is a perfluorinated biphenyl bridge connecting a rhenium-ligated imidazole or aminopropylimidazole to a distal imidazole (F8bp-im (1) and C3-F8bp-im (2)) or F (F9bp (3) and C3-F9bp (4)). All four wires bind tightly (Kd in the micromolar to nanomolar range) to the tetrahydrobiopterin-free oxidase domain of inducible nitric oxide synthase (iNOSoxy). The two fluorine-terminated wires displace water from the active site, and the two imidazole-terminated wires ligate the heme iron. Upon 355-nm excitation of iNOSoxy conjugates with 1 and 2, the active site Fe(III) is reduced to Fe(II) within 300 ps, almost 10 orders of magnitude faster than the naturally occurring reduction.