5,529 research outputs found
Kinetic Characterization and X-ray Structure of a Mutant of Haloalkane Dehalogenase with Higher Catalytic Activity and Modified Substrate Range
Conversion of halogenated aliphatics by haloalkane dehalogenase proceeds via the formation of a covalent alkyl-enzyme intermediate which is subsequently hydrolyzed by water. In the wild type enzyme, the slowest step for both 1,2-dichloroethane and 1,2-dibromoethane conversion is a unimolecular enzyme isomerization preceding rapid halide dissociation. Phenylalanine 172 is located in a helix-loop-helix structure that covers the active site cavity of the enzyme, interacts with the ClÎČ of 1,2-dichloroethane during catalysis, and could be involved in stabilization of this helix-loop-helix region of the cap domain of the enzyme. To obtain more information about the role of this residue in dehalogenase function, we performed a mutational analysis of position 172 and studied the kinetics and X-ray structure of the Phe172Trp enzyme. The Phe172Trp mutant had a 10-fold higher kcat/Km for 1-chlorohexane and a 2-fold higher kcat for 1,2-dibromoethane than the wild-type enzyme. The X-ray structure of the Phe172Trp enzyme showed a local conformational change in the helix-loop-helix region that covers the active site. This could explain the elevated activity for 1-chlorohexane of the Phe172Trp enzyme, since it allows this large substrate to bind more easily in the active site cavity. Pre-steady-state kinetic analysis showed that the increase in kcat found for 1,2-dibromoethane conversion could be attributed to an increase in the rate of an enzyme isomerization step that preceeds halide release. The observed conformational difference between the helix-loop-helix structures of the wild-type enzyme and the faster mutant suggests that the isomerization required for halide release could be a conformational change that takes place in this region of the cap domain of the dehalogenase. It is proposed that Phe172 is involved in stabilization of the helix-loop-helix structure that covers the active site of the enzyme and creates a rigid hydrophobic cavity for small apolar halogenated alkanes.
Changes in the tear proteins of diabetic patients
BACKGROUND: Previous studies have shown a significant increase in tear protein peaks in the tears of diabetic patients suffering from dry eye. The aim of this study was to analyze the tear protein patterns from patients with diabetes mellitus who do not suffer from ocular surface diseases (DIA). METHODS: A total of 515 patients were examined in this study (255 healthy subjects (controls) and 260 patients suffering from diabetes mellitus). Tear proteins were separated by sodium-dodecyl-sulfate polyacrylamide gel electrophoresis. After digital image analysis densitometric data files were created and subsequently used for multivariate statistical procedures. RESULTS: A significant increase in the number of peaks was detected in diabetic patients compared to controls (P < 0.0003). The analysis of discriminance revealed a highly significant discrimination between diabetic patients and controls (Wilks lambda: 0.27; P < 0.000001). Furthermore, a significant difference in the protein pattern of diabetic patients could be detected between those suffering from dry eye or not (P < 0.002). The changes in protein patterns of diabetic patients increased with the duration of the diabetic disease. In diabetic patients with a disease duration longer than 10 years the changes were significantly more expressed than in patients with a shorter diabetic history (P < 0.003) and in healthy subjects (P < 0.0001). CONCLUSIONS: The tear protein patterns of diabetic patients are very different in the number and intensity of spots from those of healthy subjects. Furthermore, it could be demonstrated that the differences found in the tear patterns of diabetic patients are not equal to those found in previous studies in patients suffering from dry-eye disease. The alterations in the diabetic tears were correlated with the duration of the diabetic disease. With longer disease, history changes in the tear protein patterns increased. With the course of the disease some protein peaks appeared that are not present in healthy persons. Our study shows that the analysis of electrophoretic tear protein patterns is a new non-invasive approach in the early diagnosis and analysis of the pathogenesis of diabetes induced ocular surface disease
Interrogation of caesium atoms in a fountain clock by a femtosecond laser microwave oscillator
A caesium fountain clock is operated utilizing a microwave oscillator that
derives its frequency stability from a stable laser by means of a fiber-laser
femtosecond frequency comb. This oscillator is based on the technology
developed for optical clocks and replaces the quartz based microwave oscillator
commonly used in fountain clocks. As a result, a significant decrease of the
frequency instability of the fountain clock is obtained, reaching 0.74E-14 at
100 s averaging time. We could demonstrate that for a significant range of
detected atom numbers the instability is limited by quantum projection noise
only, and that for the current status of this fountain clock the new microwave
source poses no limit on the achievable frequency instability.Comment: 4 pages, 4 figure
A re-analysis of the three-year WMAP temperature power spectrum and likelihood
We analyze the three-year WMAP temperature anisotropy data seeking to confirm
the power spectrum and likelihoods published by the WMAP team. We apply five
independent implementations of four algorithms to the power spectrum estimation
and two implementations to the parameter estimation. Our single most important
result is that we broadly confirm the WMAP power spectrum and analysis. Still,
we do find two small but potentially important discrepancies: On large angular
scales there is a small power excess in the WMAP spectrum (5-10% at l<~30)
primarily due to likelihood approximation issues between 13 <= l <~30. On small
angular scales there is a systematic difference between the V- and W-band
spectra (few percent at l>~300). Recently, the latter discrepancy was explained
by Huffenberger et al. (2006) in terms of over-subtraction of unresolved point
sources. As far as the low-l bias is concerned, most parameters are affected by
a few tenths of a sigma. The most important effect is seen in n_s. For the
combination of WMAP, Acbar and BOOMERanG, the significance of n_s =/ 1 drops
from ~2.7 sigma to ~2.3 sigma when correcting for this bias. We propose a few
simple improvements to the low-l WMAP likelihood code, and introduce two
important extensions to the Gibbs sampling method that allows for proper
sampling of the low signal-to-noise regime. Finally, we make the products from
the Gibbs sampling analysis publically available, thereby providing a fast and
simple route to the exact likelihood without the need of expensive matrix
inversions.Comment: 14 pages, 7 figures. Accepted for publication in ApJ. Numerical
results unchanged, but interpretation sharpened: Likelihood approximation
issues at l=13-30 far more important than potential foreground issues at l <=
12. Gibbs products (spectrum and sky samples, and "easy-to-use" likelihood
module) available from http://www.astro.uio.no/~hke/ under "Research
Emplacement and high-temperature evolution of gabbros of the 16.5°N oceanic core complexes (Mid-Atlantic Ridge): Insights ito the compositional variability of the lower oceanic crust
This study reports the composition of the oceanic crust from the 16.5°N region of the MidâAtlantic Ridge, a spreading ridge segment characterized by a complex detachment fault system and three main oceanic core complexes (southern, central, and northern OCCs). Lithologies recovered from the core complexes include both greenschist facies and weathered pillow basalt, diabase, peridotite, and gabbro, while only weathered and fresh pillow basalt was dredged from the rift valley floor. The gabbros are compositionally bimodal, with the magmatic crust in the region formed by scattered intrusions of chemically primitive plutonic rocks (i.e., dunites and troctolites), associated with evolved oxideâbearing gabbros. We use thermodynamic models to infer that this distribution is expected in regions where small gabbroic bodies are intruded into mantle peridotites. The occurrence of ephemeral magma chambers located in the lithospheric mantle enables large proportions of the melt to be erupted after relatively low degrees of fractionation. A large proportion of the dredged gabbros reveal evidence for deformation at highâtemperature conditions. In particular, chemical changes in response to deformation and the occurrence of very highâtemperature ultramylonites (>1000 °C) suggest that the deformation related to the oceanic detachment commenced at nearâsolidus conditions. This event was likely associated with the expulsion of interstitial, evolved magmas from the crystal mush, a mechanism that enhanced the formation of disconnected oxideâgabbro seams or layers often associated with crystalâplastic fabrics in the host gabbros. This granuliteâgrade event was soon followed by hydrothermal alteration revealed by the formation of amphiboleârich veins at highâtemperature conditions (~900 °C)
Assessment of the use of partitioning and interfacial tracers to determine the content and mass removal rates of nonaqueous phase liquids
It was assessed whether partitioning and interfacial tracers can be used to determine the content and mass removal rate of nonaqueous phase liquid (NAPL) in porous media. Retardation factors for these tracers were determined for five different model matrices contaminated with hexadecane as NAPL. The retardation of the partitioning tracer 2,4-dimethyl-3-pentanol was correlated with the degree of NAPL saturation for four of the five matrices (r(2) = 0.93, n = 8), The observed retardation factors matched the retardation factors predicted with the independently determined hexadecane-water partitioning constant and the degree of NAPL saturation, indicating that this tracer may be used to estimate the degree of NAPL saturation of porous media. The mass removal rates of NAPL from columns packed with different matrices were determined by measuring the amount of hexadecane in the column effluent during elution with electrolyte solution. These removal rates differed over 3 orders of magnitude, dependent on the matrix used. The retardation of the interfacial tracer alkylbenzenesulfonate was higher for matrices with higher NAPL mass removal rates but was not correlated to the degree of NAPL saturation. This indicates that the retardation factors of alkylbenzenesulfonates in NAPL-contaminated media contain information related to the NAPL mass removal rates
Resonances in rotationally inelastic scattering of OH() with helium and neon
We present detailed calculations on resonances in rotationally and spin-orbit
inelastic scattering of OH (X\,^2\Pi, j=3/2, F_1, f) radicals with He and Ne
atoms. We calculate new \emph{ab initio} potential energy surfaces for OH-He,
and the cross sections derived from these surfaces compare favorably with the
recent crossed beam scattering experiment of Kirste \emph{et al.} [Phys. Rev. A
\textbf{82}, 042717 (2010)]. We identify both shape and Feshbach resonances in
the integral and differential state-to-state scattering cross sections, and we
discuss the prospects for experimentally observing scattering resonances using
Stark decelerated beams of OH radicals.Comment: 14 pages, 15 Figure
Blending of nanoscale and microscale in uniform large-area sculptured thin-film architectures
The combination of large thickness ( m), large--area uniformity (75
mm diameter), high growth rate (up to 0.4 m/min) in assemblies of
complex--shaped nanowires on lithographically defined patterns has been
achieved for the first time. The nanoscale and the microscale have thus been
blended together in sculptured thin films with transverse architectures.
SiO () nanowires were grown by electron--beam evaporation onto
silicon substrates both with and without photoresist lines (1--D arrays) and
checkerboard (2--D arrays) patterns. Atomic self--shadowing due to
oblique--angle deposition enables the nanowires to grow continuously, to change
direction abruptly, and to maintain constant cross--sectional diameter. The
selective growth of nanowire assemblies on the top surfaces of both 1--D and
2--D arrays can be understood and predicted using simple geometrical shadowing
equations.Comment: 17 pages, 9 figure
Small Is Beautiful: Why Profundaplasty Should Not Be Forgotten
Background: Surgical profundaplasty (SP)is used mainly as an adjunct to endovascular management of peripheral vascular disease (PAD) today. Results from earlier series of profundaplasty alone have been controversial, especially regarding its hemodynamic effect. The question is: Can profundaplasty alone still be useful? Our aim was to evaluate its role in the modern management of vascular patients. Methods: This was a retrospective outcome study. A consecutive series of 97 patients (106 legs) from January 2000 through December 2003 were included. In 55 (52%) legs, the superficial femoral artery was occluded. These patients were included in the current analysis. Of these patients 14 (25%) were female. Mean age was 71 ((11) years. Nineteen (35%) were diabetic. The indication for operation was claudication in 29 (53%), critical leg ischemia (CLI) in 26 (47%), either with rest pain in 17 (31%), or ulcer/gangrene in 9 (16%). Endarterectomy with patch angioplasty with bovine pericardium was performed in all cases. Mean follow-up was 33(14 months. Mean preoperative ankle brachial index (ABI) was 0.6. Sustained clinical efficacy was defined as upward shift of 1 or greater on the Rutherford scale without repeat target limb revascularization (TLR) or amputation. Mortality, morbidity, need for TLR, or amputation were separate endpoints. Results: Postoperatively, ABI was significantly improved (mean=0.7), in 24 (44%) by more than 0.15. At three years, cumulative clinical success rate was 80%. Overall, patients with claudication had a better outcome than those with CLI (p=0.04). Two (4%) major amputations and 2 (4%) minor ones were performed, all in patients with CLI. None of the 9 (16%) ulcers healed. Conclusion: Profundaplasty is still a valuable option for patients with femoral PAD and claudication without tissue loss. It is a straightforward procedure that combines good efficacy with low complication rates. Further endovascular treatment may be facilitated. It is not useful for patients with the combination of critical ischemia and tissue los
Electron localization by a magnetic vortex
We study the problem of an electron in two dimensions in the presence of a
magnetic vortex with a step-like profile. Dependending on the values of the
effective mass and gyromagnetic factor of the electron, it may be trapped by
the vortex. The bound state spectrum is obtained numerically, and some limiting
cases are treated analytically.Comment: 8 pages, latex, 4 figure
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