Electron transfer in acetohydroxy acid synthase as a side reaction of catalysis. implications for the reactivity and partitioning of the carbanion/enamine form of (alpha-hydroxyethyl)thiamin diphosphate in a "nonredox" flavoenzyme

Acetohydroxy acid synthases (AHAS) are thiamin diphosphate- (ThDP-) and FAD-dependent enzymes that catalyze the first common step of branched-chain amino acid biosynthesis in plants, bacteria, and fungi. Although the flavin cofactor is not chemically involved in the physiological reaction of AHAS, it has been shown to be essential for the structural integrity and activity of the enzyme. Here, we report that the enzyme-bound FAD in AHAS is reduced in the course of catalysis in a side reaction. The reduction of the enzyme-bound flavin during turnover of different substrates under aerobic and anaerobic conditions was characterized by stopped-flow kinetics using the intrinsic FAD absorbance. Reduction of enzyme-bound FAD proceeds with a net rate constant of k' = 0.2 s(-1) in the presence of oxygen and approximately 1 s(-1) under anaerobic conditions. No transient flavin radicals are detectable during the reduction process while time-resolved absorbance spectra are recorded. Reconstitution of the binary enzyme-FAD complex with the chemically synthesized intermediate 2-(hydroxyethyl)-ThDP also results in a reduction of the flavin. These data provide evidence for the first time that the key catalytic intermediate 2-(hydroxyethyl)ThDP in the carbanionic/enamine form is not only subject to covalent addition of 2-keto acids and an oxygenase side reaction but also transfers electrons to the adjacent FAD in an intramolecular redox reaction yielding 2-acetyl-ThDP and reduced FAD. The detection of the electron transfer supports the idea of a common ancestor of acetohydroxy acid synthase and pyruvate oxidase, a homologous ThDP- and FAD-dependent enzyme that, in contrast to AHASs, catalyzes a reaction that relies on intercofactor electron transfer.

Time and space-resolved measurement of a gas-puff laser-plasma x-ray source

The dynamic interaction processes between a nano-second laser pulse and a gas-puff target, such as those of plasma formation, laser heating, and x-ray emission, have been investigated quantitatively. Time and space-resolved x-ray and optical measurement techniques were used in order to investigate time-resolved laser absorption and subsequent x-ray generation. Efficient absorption of the incident laser energy into the gas-puff target of 17%, 12%, 38%, and 91% for neon, argon, krypton, and xenon, respectively, was shown experimentally. It was found that the laser absorption starts and, simultaneously, soft x-ray emission occurs. The soft x-ray lasts much longer than the laser pulse due to the recombination. Temporal evolution of the soft x-ray emission region was analyzed by comparing the experimental results to the results of the model calculation, in which the laser light propagation through a gas-puff plasma was taken into account. (C) 2003 American Institute of Physics.

Combined time- and space-resolved Raman spectrometer for the non-invasive depth profiling of chemical hazards

A time-resolved inverse spatially offset Raman spectrometer was constructed for depth profiling of Raman-active substances under both the lab and the field environments. The system operating principles and performance are discussed along with its advantages relative to traditional continuous wave spatially offset Raman spectrometer. The developed spectrometer uses a combination of space- and time-resolved detection in order to obtain high-quality Raman spectra from substances hidden behind coloured opaque surface layers, such as plastic and garments, with a single measurement. The time-gated spatially offset Raman spectrometer was successfully used to detect concealed explosives and drug precursors under incandescent and fluorescent background light as well as under daylight. The average screening time was 50 s per measurement. The excitation energy requirements were relatively low (20 mW) which makes the probe safe for screening hazardous substances. The unit has been designed with nanosecond laser excitation and gated detection, making it of lower cost and complexity than previous picosecond-based systems, to provide a functional platform for in-line or in-field sensing of chemical substances.

Stable Temperature Characteristics of InAs/GaAs Quantum Dots at Long Wavelength Emission

The time-resolved photoluminescence and steady photoluminescence (TRPL and PL) spectra on self-assembled InAs/GaAs quantum dots (QDs) are investigated. By depositing GaAs/InAs short period superlattices (SLs), 1. 48 μtm emission is obtained at room temperature. Temperature dependent PL measurements show that the PL intensity of the emission is very steady. It decays only to half as the temperature increases from 15 K to room temperature, while at the same time, the intensity of the other emission decreases by a factor of 5 orders of magnitude. These two emissions are attributed to large-size QDs and short period superlattices (SLs), respectively. Large-size QDs are easier to capture and confine carriers,which benefits the lifetime of PL, and therefore makes the emission intensity insensitive to the temperature.

Emission lines of Fe X in active region spectra obtained with the Solar Extreme-ultraviolet Research Telescope and Spectrograph

Fully relativistic calculations of radiative rates and electron impact excitation cross-sections for Fe X are used to derive theoretical emission-line ratios involving transitions in the 174-366 angstrom wavelength range. A comparison of these with solar active region observations obtained during the 1989 and 1995 flights of the Solar Extreme-ultraviolet Research Telescope and Spectrograph (SERTS) reveals generally very good agreement between theory and experiment. Several Fe X emission features are detected for the first time in SERTS spectra, while the 3s(2)3p(5) P-2(3/2)-3s(2)3p(4)(S-1)3d D-2(3/2) transition at 195.32 angstrom is identified for the first time (to our knowledge) in an astronomical source. The most useful Fe X electron density (N-e) diagnostic line ratios are assessed to be 175.27/174.53 and 175.27/177.24, which both involve lines close in wavelength and free from blends, vary by factors of 13 between N-e = 10(8) and 10(11) cm(-3), and yet show little temperature sensitivity. Should these lines not be available, then the 257.25/345.74 ratio may be employed to determine N-e, although this requires an accurate evaluation of the instrument intensity calibration over a relatively large wavelength range. However, if the weak 324.73 angstrom line of Fe X is reliably detected, the use of 324.73/345.74 or 257.25/324.73 is recommended over 257.25/345.74. Electron densities deduced from 175.27/174.53 and 175.27/177.24 for the stars Procyon and alpha Cen, using observations from the Extreme-Ultraviolet Explorer (EUVE) satellite, are found to be consistent and in agreement with the values of N-e determined from other diagnostic ratios in the EUVE spectra. A comparison of several theoretical extreme-ultraviolet Fe X line ratios with experimental values for a theta-pinch, for which the plasma parameters have been independently determined, reveals reasonable agreement between theory and observation, providing some independent support for the accuracy of the adopted atomic data.

Emission lines of Fe XI in the 257-407 A wavelength region observed in solar spectra from EIS/Hinode and SERTS

Theoretical emission-line ratios involving Fe xi transitions in the 257-407 A wavelength range are derived using fully relativistic calculations of radiative rates and electron impact excitation cross-sections. These are subsequently compared with both long wavelength channel Extreme-Ultraviolet Imaging Spectrometer (EIS) spectra from the Hinode satellite (covering 245-291 A) and first-order observations (similar to 235-449 A) obtained by the Solar Extreme-ultraviolet Research Telescope and Spectrograph (SERTS). The 266.39, 266.60 and 276.36 A lines of Fe xi are detected in two EIS spectra, confirming earlier identifications of these features, and 276.36 A is found to provide an electron density (N-e) diagnostic when ratioed against the 257.55 A transition. Agreement between theory and observation is found to be generally good for the SERTS data sets, with discrepancies normally being due to known line blends, while the 257.55 A feature is detected for the first time in SERTS spectra. The most useful Fe xi electron density diagnostic is found to be the 308.54/352.67 intensity ratio, which varies by a factor of 8.4 between N-e = 108 and 1011 cm-3, while showing little temperature sensitivity. However, the 349.04/352.67 ratio potentially provides a superior diagnostic, as it involves lines which are closer in wavelength, and varies by a factor of 14.7 between N-e = 108 and 1011 cm-3. Unfortunately, the 349.04 A line is relatively weak, and also blended with the second-order Fe x 174.52 A feature, unless the first-order instrument response is enhanced.

Prompt electron emission and collisional ionization of ambient gas during pulsed laser ablation of silver

A silver target kept under partial vacuum conditions was irradiated with focused nanosecond pulses at 1:06 mm from a Nd:YAG laser. The electron emission monitored with a Langmuir probe shows a clear twin-peak distribution. The first peak which is very sharp has only a small delay and it indicates prompt electron emission with energy as much as 60 5 eV. Also the prompt electron emission shows a temporal profile with a width that is same as that for the laser pulse whereas the second peak is broader, covers several microseconds, and represents the low-energy electrons (2 0:5 eV) associated with the laser-induced silver plasma as revealed by time-of-flight measurements. It has been found that prompt electrons ejected from the target collisionally excite and ionize ambient gas molecules. Clearly resolved rotational structure is observed in the emission spectra of ambient nitrogen molecules. Combined with time-resolved spectroscopy, the prompt electrons can be used as excitation sources for various collisional excitation–relaxation experiments. The electron density corresponding to the first peak is estimated to be of the order of 1017 cm?--3 and it is found that the density increases as a function of distance away from the target. Dependence of probe current on laser intensity shows plasma shielding at high laser intensities.

Emission characteristics and dynamics of C2 from laser produced graphite plasma

The emission features of laser ablated graphite plume generated in a helium ambient atmosphere have been investigated with time and space resolved plasma diagnostic technique. Time resolved optical emission spectroscopy is employed to reveal the velocity distribution of different species ejected during ablation. At lower values of laser fluences only a slowly propagating component of C2 is seen. At high fluences emission from C2 shows a twin peak distribution in time. The formation of an emission peak with diminished time delay giving an energetic peak at higher laser fluences is attributed to many body recombination. It is also observed that these double peaks get modified into triple peak time of flight distribution at distances greater than 16 mm from the target. The occurrence of multiple peaks in the C2 emission is mainly due to the delays caused from the different formation mechanism of C2 species. The velocity distribution of the faster peak exhibits an oscillating character with distance from the target surface.

Analysis and control of transient spectra using time-dependent density functional theory.

215 p.

Spectrally Resolved Resonance Energy Transfer from ZnO:MgO Nanocrystals

Resonance energy transfer (RET) from the visible emission of core−shell ZnO:MgO nanocrystals to Nile Red chromophores, following band gap excitation in the UV, has been investigated for four different nanocrystal sizes. With use of steady state and time-resolved fluorescence spectroscopic measurements the wavelength dependent RET efficiencies have been determined. The RET process in ZnO:MgO nanocrystals occurs from emissions involving trap state recombination. There are two such processes with different RET efficiencies for the same particle size. This is shown to be a consequence of the fact that the recombination processes giving rise to the two emissions are located at different distances from the center of the particle so that the donor−acceptor distances for the two are different, even for the same particle size.

Possibility of lifetime measurement in a flowing medium with phase fluorometry

The possibility of lifetime measurement in a flowing medium with phase fluorometry is investigated theoretically. A 3-D time dependent partial differential equation of the number density of atoms (or molecules) in the upper level of the fluorescence transition is solved analytically, taking flow, diffusion, optical excitation, decay, Doppler shift, and thickness of the excitation light sheet into account. An analytical expression of the intensity of the fluorescence signal in the flowing medium is deduced. Conditions are given, in which the principle of lifetime measurement with phase fluorometry in the static sample cell can be used in a flowing medium.

Particulate matter emission during start-up and transient operation of a spark-ignition engine

In order to understand why emissions of Particulate Matter (PM) from Spark-Ignition (SI) automobiles peak during periods of transient operation such as rapid accelerations, a study of controlled, repeatable transients was performed. Time-resolved engine-out PM emissions from a modern four-cylinder engine during transient load and air/fuel ratio operation were examined, and the results could be fit in most cases to a first order time response. The time constants for the transient response are similar to those measured for changes in intake valve temperature, reflecting the strong dependence of PM emissions on the amount of liquid fuel in the combustion chamber. In only one unrepeatable case did the time response differ from a first order function: showing an overshoot in PM emissions during transition from the initial to the final steady state PM emission level. PM emissions during controlled, motored start-up experiments show a peak at start-up followed by a period during which emissions are either relatively constant or drift somewhat. When the fuel injection and ignition are shut off, PM emissions also peak briefly, but rapidly decay to low levels. Qualitative implications on the study and modeling of PM emissions during transient engine operation are discussed. Copyright © 1999 Society of Automotive Engineers, Inc.

Abnormal energy dependence of photoluminescence decay time in InGaN epilayer

We employ photoluminescence (PL) and time-resolved PL to study exciton localization effect in InGaN epilayers. By measuring the exciton decay time as a, function of the monitored emission energy at different temperatures, we have found unusual behaviour of the energy dependence in the PL decay process. At low temperature, the measured PL decay time increases with the emission energy. It decreases with the emission energy at 200K, and remains nearly constant at the intermediate temperature of 120K. We have studied the dot size effect on the radiative recombination time by calculating the temperature dependence of the exciton recombination lifetime in quantum dots, and have found that the observed behaviour can be well correlated to the exciton localization in quantum dots. This suggestion is further supported by steady state PL results.

Photoluminescence properties of a GaN0.015As0.985/GaAs single quantum well under short pulse excitation

Under short pulse laser excitation, we have observed an extra high-energy photoluminescence (PL) emission from GaNAs/GaAs single quantum wells (QWs). It dominates the PL spectra under high excitation and/or at high temperature. By measuring the PL dependence on both temperature and excitation power and by analyzing the time-resolved PL results, we have attributed the PL peak to the recombination of delocalized excitons in QWs. Furthermore, a competition process between localized and delocalized excitons is observed in the temperature-dependent PL spectra under the short pulse excitation. This competition is believed to be responsible for the temperature-induced S-shaped PL shift often observed in the disordered alloy semiconductor system under continuous-wave excitation. (C) 2001 American Institute of Physics.

Two-dimensional excitonic emission in InAs submonolayers

Photoluminescence (PL) and time-resolved photoluminescence (TRPL) were used to study optical emissions of ultrathin InAs layers with average layer thickness ranging from 1/12 to 1 ML grown on GaAs substrates. We have found that the inhomogeneous broadening of the PL from InAs layers can be well described by the quantum-well model with InAs islands coupling to each other and being regarded as a quasiwell. From the temperature dependence of the exciton linewidth, the exciton-LO-phonon scattering coefficient was found to be comparable to that in conventional two-dimensional quantum wells. In the TRPL measurements, the PL decay time increases linearly with temperature, which is a typical characteristic of free excitons in quantum wells. All these results indicate that the excitons localized in InAs exhibit two-dimensional properties of quantum wells, despite the topographical islandlike structure.