338 research outputs found
Rapport Commissie rechtseenheid bestuursrecht: differentiatie in rechtseenheidsvoorzieningen
The Legitimacy and Effectiveness of Law & Governance in a World of Multilevel Jurisdiction
Polarization-controlled optimal scatter suppression in transient absorption spectroscopy
Ultrafast transient absorption spectroscopy is a powerful technique to study fast photo-induced processes, such as electron, proton and energy transfer, isomerization and molecular dynamics, in a diverse range of samples, including solid state materials and proteins. Many such experiments suffer from signal distortion by scattered excitation light, in particular close to the excitation (pump) frequency. Scattered light can be effectively suppressed by a polarizer oriented perpendicular to the excitation polarization and positioned behind the sample in the optical path of the probe beam. However, this introduces anisotropic polarization contributions into the recorded signal. We present an approach based on setting specific polarizations of the pump and probe pulses, combined with a polarizer behind the sample. Together, this controls the signal-to-scatter ratio (SSR), while maintaining isotropic signal. We present SSR for the full range of polarizations and analytically derive the optimal configuration at angles of 40.5° between probe and pump and of 66.9° between polarizer and pump polarizations. This improves SSR by 33 52 ≈. (or 3 compared to polarizer parallel to probe). The calculations are validated by transient absorption experiments on the common fluorescent dye Rhodamine B. This approach provides a simple method to considerably improve the SSR in transient absorption spectroscopy
Femtosecond Stimulated Raman Study of the Photoactive Flavoprotein AppABLUF
Femtosecond stimulated Raman Spectroscopy (FSRS) is applied to study the photocycle of a blue light using flavin (BLUF) domain photoreceptor, AppABLUF. It is shown that FSRS spectra are sensitive to the light adapted state of the protein and probe its excited state dynamics. The dominant contribution to the most sensitive excited state Raman active modes is from flavin ring modes. However, TD-DFT calculations for excited state structures indicate that reproduction and assignment of the experimentally observed spectral shift will require high level calculations on the flavin in its specific protein environment
Ultrafast transient absorption spectroscopy: principles and application to photosynthetic systems
The photophysical and photochemical reactions, after light absorption by a photosynthetic pigment–protein complex, are among the fastest events in biology, taking place on timescales ranging from tens of femtoseconds to a few nanoseconds. The advent of ultrafast laser systems that produce pulses with femtosecond duration opened up a new area of research and enabled investigation of these photophysical and photochemical reactions in real time. Here, we provide a basic description of the ultrafast transient absorption technique, the laser and wavelength-conversion equipment, the transient absorption setup, and the collection of transient absorption data. Recent applications of ultrafast transient absorption spectroscopy on systems with increasing degree of complexity, from biomimetic light-harvesting systems to natural light-harvesting antennas, are presented. In particular, we will discuss, in this educational review, how a molecular understanding of the light-harvesting and photoprotective functions of carotenoids in photosynthesis is accomplished through the application of ultrafast transient absorption spectroscopy
Excitons in a Photosynthetic Light-Harvesting System: A Combined Molecular Dynamics/Quantum Chemistry and Polaron Model Study
The dynamics of pigment-pigment and pigment-protein interactions in
light-harvesting complexes is studied with a novel approach which combines
molecular dynamics (MD) simulations with quantum chemistry (QC) calculations.
The MD simulations of an LH-II complex, solvated and embedded in a lipid
bilayer at physiological conditions (with total system size of 87,055 atoms)
revealed a pathway of a water molecule into the B800 binding site, as well as
increased dimerization within the B850 BChl ring, as compared to the
dimerization found for the crystal structure. The fluctuations of pigment (B850
BChl) excitation energies, as a function of time, were determined via ab initio
QC calculations based on the geometries that emerged from the MD simulations.
From the results of these calculations we constructed a time-dependent
Hamiltonian of the B850 exciton system from which we determined the linear
absorption spectrum. Finally, a polaron model is introduced to describe quantum
mechanically both the excitonic and vibrational (phonon) degrees of freedom.
The exciton-phonon coupling that enters into the polaron model, and the
corresponding phonon spectral function are derived from the MD/QC simulations.
It is demonstrated that, in the framework of the polaron model, the absorption
spectrum of the B850 excitons can be calculated from the autocorrelation
function of the excitation energies of individual BChls, which is readily
available from the combined MD/QC simulations. The obtained result is in good
agreement with the experimentally measured absorption spectrum.Comment: REVTeX3.1, 23 pages, 13 (EPS) figures included. A high quality PDF
file of the paper is available at
http://www.ks.uiuc.edu/Publications/Papers/PDF/DAMJ2001/DAMJ2001.pd
BLUF Domain Function Does Not Require a Metastable Radical Intermediate State
BLUF
(blue light using flavin) domain proteins are an important
family of blue light-sensing proteins which control a wide variety
of functions in cells. The primary light-activated step in the BLUF
domain is not yet established. A number of experimental and theoretical
studies points to a role for photoinduced electron transfer (PET)
between a highly conserved tyrosine and the flavin chromophore to
form a radical intermediate state. Here we investigate the role of
PET in three different BLUF proteins, using ultrafast broadband transient
infrared spectroscopy. We characterize and identify infrared active
marker modes for excited and ground state species and use them to
record photochemical dynamics in the proteins. We also generate mutants
which unambiguously show PET and, through isotope labeling of the
protein and the chromophore, are able to assign modes characteristic
of both flavin and protein radical states. We find that these radical
intermediates are not observed in two of the three BLUF domains studied,
casting doubt on the importance of the formation of a population of
radical intermediates in the BLUF photocycle. Further, unnatural amino
acid mutagenesis is used to replace the conserved tyrosine with fluorotyrosines,
thus modifying the driving force for the proposed electron transfer
reaction; the rate changes observed are also not consistent with a
PET mechanism. Thus, while intermediates of PET reactions can be observed
in BLUF proteins they are not correlated with photoactivity, suggesting
that radical intermediates are not central to their operation. Alternative
nonradical pathways including a keto–enol tautomerization induced
by electronic excitation of the flavin ring are considered
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The reinforcement sensitivity theory of personality in children: A new questionnaire
We report the development of a self-report questionnaire of the reinforcement sensitivity theory (RST) of personality for use with children. Focus groups were held with children to sample their experiences of situations modelled on components of three RST systems: fight-flight-freeze system (FFFS, related to fear), behavioural inhibition system (BIS, related to anxiety), and behavioural approach system (BAS, related to approach). The thematic responses formed the conceptual anchors to the development of test items that were examined using exploratory factor analysis in a sample of 288 9–13 year olds. After eliminating items that did not load on their designated factor, or substantially cross-loaded over factors, the original 48 items were reduced to 21 items: 7 items for each of the BIS, FFFS and BAS factors. The separation of the BIS and FFFS items across two factors is consistent with the revised model of RST. We offer this new questionnaire as a RST measure of fundamental motivation and emotion traits in children
Nanoscale Confinement and Fluorescence Effects of Bacterial Light Harvesting Complex LH2 in Mesoporous Silicas
Many key chemical and biochemical reactions, particularly in living cells, take place in confined space at the mesoscopic scale. Toward understanding of physicochemical nature of biomacromolecules confined in nanoscale space, in this work we have elucidated fluorescence effects of a light harvesting complex LH2 in nanoscale chemical environments. Mesoporous silicas (SBA-15 family) with different shapes and pore sizes were synthesized and used to create nanoscale biomimetic environments for molecular confinement of LH2. A combination of UV-vis absorption, wide-field fluorescence microscopy, and in situ ellipsometry supports that the LH2 complexes are located inside the silica nanopores. Systematic fluorescence effects were observed and depend on degree of space confinement. In particular, the temperature dependence of the steady-state fluorescence spectra was analyzed in detail using condensed matter band shape theories. Systematic electronic-vibrational coupling differences in the LH2 transitions between the free and confined states are found, most likely responsible for the fluorescence effects experimentally observed
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