Coexistence of domains with distinct order and polarity in fluid bacterial membranes

In this study we sought the detection and characterization of bacterial membrane domains. Fluorescence generalized polarization (GP) spectra of laurdan-labeled Escherichia coli and temperature dependencies of both laurdan's GP and fluorescence anisotropy of 1,3-diphenyl-1,3,5-hexatriene (DPH) (rDPH) affirmed that at physiological temperatures, the E. coli membrane is in a liquid-crystalline phase. However, the strong excitation wavelength dependence of rlaurdan at 37 degrees C reflects membrane heterogeneity. Time-resolved fluorescence emission spectra, which display distinct biphasic redshift kinetics, verified the coexistence of two subpopulations of laurdan. In the initial phase, <50 ps, the redshift in the spectral mass center is much faster for laurdan excited at the blue edge (350 nm), whereas at longer time intervals, similar kinetics is observed upon excitation at either blue or red edge (400 nm). Excitation in the blue region selects laurdan molecules presumably located in a lipid domain in which fast intramolecular relaxation and low anisotropy characterize laurdan's emission. In the proteo-lipid domain, laurdan motion and conformation are restricted as exhibited by a slower relaxation rate, higher anisotropy and a lower GP value. Triple-Gaussian decomposition of laurdan emission spectra showed a sharp phase transition in the temperature dependence of individual components when excited in the blue but not in the red region. At least two kinds of domains of distinct polarity and order are suggested to coexist in the liquid-crystalline bacterial membrane: a lipid-enriched and a proteolipid domain. In bacteria with chloramphenicol (Cam)-inhibited protein synthesis, laurdan showed reduced polarity and restoration of an isoemissive point in the temperature-dependent spectra. These results suggest a decrease in membrane heterogeneity caused by Cam-induced domain dissipation.     

Photoinduced decomposition of the polybromostyryl carbanions

The photoinduced decay in THF of the sodium salt of the polybromostyryl carbanions (PS_B?) was investigated in vacuum at ?78°C. Maximal rates of decay were obtained when the wavelength of the incident light corresponded to the absorption maximum of the PS_B? at 495 nm. The quantum yield of the process was found to be 13 eq./Einstein. Products of the photostimulated decompositions at ?78°C were found to be different from those obtained as result of the “dark” thermal decomposition at –20°C. Decomposition of the PS_B? carbanions was accompanied by evolution of Br^? anions and by formation of polyaryl carbanions absorbing in the 270- to 400-nm range. The mechanism of the photoinduced decomposition of PS_B?, though basically analogous to the S_RN1 reaction mechanism, was found to differ significantly from the latter. The intramolecular dissociative electron transfer was assumed to represent its initiation step. Its propagation was attributed to the addition of PS_B? to the initially formed biradicals, ejection of Br^? from the resulting radical anions, and intramolecular electron transfer yielding polybromostyryl radicals. Products of the photoinduced decomposition of PS_B? remained soluble in THF when the intensity of the incident light was relatively weak, though their molecular weight distribution broadened. Insoluble, highly crosslinked polymers were obtained when the intensity of the incident light was very high.     

Identification of phenylbutyrylglutamine,a new metabolite of phenylbutyrate metabolism in humans

Phenylbutyrate is used in humans for treating inborn errors of ureagenesis, certain forms of cancer, cystic fibrosis and thalassemia. The known metabolism of phenylbutyrate leads to phenylacetylglutamine, which is excreted in urine. We have identified phenylbutyrylglutamine as a new metabolite of phenylbutyrate in human plasma and urine. We describe the synthesis of phenylbutyrylglutamine and its assay by gas chromatography/mass spectrometry as a tert-butyldimethylsilyl or methyl derivative, using standards of [(2)H(5)]phenylbutyrylglutamine and phenylpropionylglutamine. After administration of phenylbutyrate to normal humans, the cumulative urinary excretion of phenylacetate, phenylbutyrate, phenylacetylglutamine and phenylbutyrylglutamine amounts to about half of the dose of phenylbutyrate. Thus, additional metabolites of phenylbutyrate are yet to be identified.     

Fluorescence resonance energy transfer in CdSe/ZnS-DNA conjugates: probing hybridization and DNA cleavage

Nucleic-acid-functionalized CdSe/ZnS quantum dots (QDs) were hybridized with the complementary Texas-Red-functionalized nucleic acid. The hybridization was monitored by following the fluorescence resonance energy transfer from the QDs to the dye units. Treatment of the QD/dye DNA duplex structure with DNase I resulted in the cleavage of the DNA and the recovery of the fluorescence properties of the CdSe/ZnS QDs. The luminescence properties of the QDs were, however, only partially recovered due to the nonspecific adsorption of the dye onto the QDs. Similarly, nucleic-acid-functionalized Au nanoparticles (Au NPs) were hybridized with the complementary Texas-Red-labeled nucleic acid. The hybridization was followed by the fluorescence quenching of the dye by the Au NPs. Treatment of the Au NP/dye DNA duplex with DNase I resulted in the cleavage of the DNA and the partial recovery of the dye fluorescence. The incomplete recovery of the dye fluorescence originated from the nonspecific binding of the dye units to the Au NPs. The nonspecific binding of the dye to the CdSe/ZnS QDs and the Au NPs is attributed to nonprotected surface vacancies in the two systems.     

Rate constants for the reactions of Ar+ with CO2 and SO2 as a function of temperature (300–1500 K)

We have measured the rate constants for the reactions of Ar⁺ with CO₂ and SO₂ from 300 to 1500 K in a high temperature flowing afterglow. For the reaction with CO₂, we have found that all modes of energy, i.e., translation, rotation, and vibration, affect the rate constant to the same degree up to a total energy of 0.4 eV. Above 0.4 eV total energy, internal energy decreases the rate constants more effectively than does translational energy. For the reaction of Ar⁺ with SO₂, the rate constants go through a minimum at about 900 K. By comparing our results to drift tube data, we derive rate constants for reaction from the υ=0 and υ>0 vibrational levels. At low energy, the vibrationally excited SO₂ molecules react with Ar⁺ approximately twice as fast as the ground state molecules. Both vibrational modes have similar temperature dependences.     

Characterization of an orphan Ra–Be neutron source

Journal of Radioanalytical and Nuclear Chemistry - An orphan radium-beryllium (Ra–Be) neutron source (Nuclear Chicago Corporation) detected inside a scrap metal shipping container, was seized...     

Evaluation of link-track performances using DEM

A two-dimensional discrete-element model for the interaction between link-track and soil is presented. The model was developed using commercial PFC2D code. Two different particles, sphere and clump of two spheres, were used to represent the soil. The soil parameters of the model were determined using Hertzian contact theory. Based on the model and soil parameters, simulations of biaxial tests and calculations of the internal angle of friction and cohesion were preformed. The simulation results showed that the internal angle of friction should not exceed the value of 0.65 when using the spherical particles. Based on the clumped particles model, simulations of shear tests with two grouser plates (lengths 100 and 150 mm) were performed under different soil conditions, normal pressures, and cleat heights. A curve fitting of the simulation results was performed using three semi-empirical models from Bekker, Janosi, and Wong for representing the shear stress–displacement relationship. The best fitting was achieved using Wong’s approach. The simulation results of the cleat effects were compared with Bekker’s grouser approach and McKyes’s formulation for soil–blade interaction. In most of the cases, the results of Bekker’s model were the lower bound and McKyes’s model, the upper bound of the DEM simulation results. The properties of the soil model for the DEM were determined using simulation results of shear tests by grouser plate. In the range investigated, the size of the shearing grouser plate is not significant in determining the soil model properties.     

Traction performance of a pushed/pulled drive wheel

A comprehensive method for prediction of off-road driven wheel performance is presented, assuming a parabolic wheel–soil contact surface. The traction performance of a driven wheel is predicted for both driving and braking modes. Simulations show significant non-symmetry of the traction performance of the driving and braking wheels. The braking force is significantly greater than the traction force reached in the driving mode. In order to apply the suggested model for prediction of the traction performance of a 4WD vehicle, the load transfer effect was considered. Simulated traction performances of front and rear driven wheels differ significantly, due to the load transfer. In the driving mode, the rear driven wheel develops a net traction force greater than that of the front wheel. On the other hand, in the braking mode the front driven wheel develops a braking force significantly greater than that of the rear driven wheel due to a pushed/pulled force affected by the load transfer. The suggested model was successfully verified by the data reported in literature and by full-scale field experiments with a special wheel-testing device. The developed approach may improve the prediction of off-road multi-drive vehicle traction performance.     

Retardation in rigid motion and degree of rigidity

Retardation features of rigid motion, in flat two-dimensional space-time, are interpreted in terms of a consistent picture of signals. A similar analysis is adopted to effect a natural transition from rigid to nonrigid frames for which a degree of rigidity, having a simple physical meaning, can be defined.