1,901 research outputs found
Identification and Characterization of NeuB3 from Campylobacter jejuni as a Pseudaminic Acid Synthase
Campylobacter jejuni and Campylobacter coli are the main causes of bacterial diarrhea worldwide, and Helicobacter pylori is known to cause duodenal ulcers. In all of these pathogenic organisms, the flagellin proteins are heavily glycosylated with a 2-keto-3-deoxy acid, pseudaminic acid (5,7-diacetamido-3,5,7,9-tetradeoxy-L-glycero-L-manno-nonulosonic acid). The presence of pseudaminic acid is required for the proper development of the flagella and is thereby necessary for motility in, and invasion of, the host. In this study we report the first characterization of NeuB3 from C. jejuni as a pseudaminic acid synthase; the enzyme directly responsible for the biosynthesis of pseudaminic acid. Pseudaminic acid synthase catalyzes the condensation of phosphoenolpyruvate (PEP) with the hexose, 2,4-diacetamido-2,4,6-trideoxy-L-altrose (6-deoxy-AltdiNAc), to form pseudaminic acid and phosphate. The enzymatic activity was monitored using 1H and 31P NMR spectroscopy, and the product was isolated and characterized. Kinetic analysis reveals that pseudaminic acid synthase requires the presence of a divalent metal ion for catalysis and that optimal catalysis occurs at pH 7.0. A coupled enzymatic assay gave the values for k(cat) of 0.65 +/- 0.01 s(-1), K(m)PEP of 6.5 +/- 0.4 microM, and K(m)6-deoxy-AltdiNAc of 9.5 +/- 0.7 microM. A mechanistic study on pseudaminic acid synthase, using [2-18O]PEP, shows that catalysis proceeds through a C-O bond cleavage mechanism similar to other PEP condensing synthases such as sialic acid synthase
Measurements of pernitric acid at the South Pole during ISCAT 2000
The first measurements of pernitric acid at the South Pole were performed during the second Investigation of Sulfur Chemistry in the Antarctic Troposphere (ISCAT 2000). Observed HO2NO2 concentrations averaged 25 pptv. Simple steady-state calculations constrained by measurements show that the lifetime of pernitric acid was largely controlled by dry deposition, with thermal decomposition becoming increasingly important at warmer temperatures. We determined that the pernitric acid equilibrium constant is less uncertain than indicated in the literature. One consequence of pernitric acid deposition to the snow surface is that it is an important sink for both NOx and HOx. Another is that the photochemistry of HO2NO2 in the Antarctic snowpack may be a NOx source in addition to nitrate photolysis. This might be one of the important differences in snow photochemistry between the South Pole and warmer polar sites
Subharmonic Generation in Quantum Systems
We show how the classical-quantum correspondence permits long-lived
subharmonic motion in a quantum system driven by a periodic force.
Exponentially small deviations from exact subharmonicity are due to coherent
tunneling between quantized vortex tubes which surround classical elliptic
periodic orbits.Comment: 11 pages + 5 figures (available upon request), Revtex 3.0,
NSF-ITP-93-4
Quantum oscillations and optical conductivity in Rashba spin-splitting BiTeI
We report the observation of Shubnikov-de Haas (SdH) oscillations in single
crystals of the Rashba spin-splitting compound BiTeI, from both longitudinal
() and Hall () magnetoresistance. Under magnetic field up
to 65 T, we resolved unambiguously only one frequency T,
corresponding to a Fermi momentum \AA.The
amplitude of oscillations is strongly suppressed by tilting magnetic field,
suggesting a highly two-dimensional Fermi surface. Combining with optical
spectroscopy, we show that quantum oscillations may be consistent with a bulk
conduction band having a Rashba splitting momentum \AA
Structural and Mechanistic Analysis of Sialic Acid Synthase NeuB from Neisseria meningitidis in Complex with Mn2+, Phosphoenolpyruvate, and N-Acetylmannosaminitol *
In Neisseria meningitidis and related bacterial pathogens, sialic acids play critical roles in mammalian cell immunity evasion and are synthesized by a conserved enzymatic pathway that includes sialic acid synthase (NeuB, SiaC, or SynC). NeuB catalyzes the condensation of phosphoenolpyruvate (PEP) and N-acetylmannosamine, directly forming N-acetylneuraminic acid (or sialic acid). In this paper we report the development of a coupled assay to monitor NeuB reaction kinetics and an 18O-labeling study that demonstrates the synthase operates via a C-O bond cleavage mechanism. We also report the first structure of a sialic acid synthase, that of NeuB, revealing a unique domain-swapped homodimer architecture consisting of a (beta/alpha)8 barrel (TIM barrel)-type fold at the N-terminal end and a domain with high sequence identity and structural similarity to the ice binding type III antifreeze proteins at the C-terminal end of the enzyme. We have determined the structures of NeuB in the malate-bound form and with bound PEP and the substrate analog N-acetylmannosaminitol to 1.9 and 2.2 A resolution, respectively. Typical of other TIM barrel proteins, the active site of NeuB is located in a cavity at the C-terminal end of the barrel; however, the positioning of the swapped antifreeze-like domain from the adjacent monomer provides key residues for hydrogen bonding with substrates in the active site of NeuB, a structural feature that leads to distinct modes of substrate binding from other PEP-utilizing enzymes that lack an analogous antifreeze-like domain. Our observation of a direct interaction between a highly ordered manganese and the N-acetylmannosaminitol in the NeuB active site also suggests an essential role for the ion as an electrophilic catalyst that activates the N-acetylmannosamine carbonyl to the addition of PEP
Characterization of thermal effects in the Enhanced LIGO Input Optics
We present the design and performance of the LIGO Input Optics subsystem as
implemented for the sixth science run of the LIGO interferometers. The Initial
LIGO Input Optics experienced thermal side effects when operating with 7 W
input power. We designed, built, and implemented improved versions of the Input
Optics for Enhanced LIGO, an incremental upgrade to the Initial LIGO
interferometers, designed to run with 30 W input power. At four times the power
of Initial LIGO, the Enhanced LIGO Input Optics demonstrated improved
performance including better optical isolation, less thermal drift, minimal
thermal lensing and higher optical efficiency. The success of the Input Optics
design fosters confidence for its ability to perform well in Advanced LIGO
Infrared phonon anomaly and magnetic excitations in single-crystal Cu3Bi(SeO3)2O2Cl
nfrared reflection and transmission as a function of temperature have been measured on single crystals of Cu3Bi(SeO3)2O2Cl. The complex dielectric function and optical properties along all three principal axes of the orthorhombic cell were obtained via Kramers-Kronig analysis and by fits to a Drude-Lorentz model. Below 115 K, 16 additional modes [8(Eâ„aÌ)+6(Eâ„bÌ)+2(Eâ„cÌ)] appear in the phonon spectra; however, powder x-ray diffraction measurements do not detect a new structure at 85 K. Potential explanations for the new phonon modes are discussed. Transmission in the far infrared as a function of temperature has revealed magnetic excitations originating below the magnetic ordering temperature (TcâŒ24 K). The origin of the excitations in the magnetically ordered state will be discussed in terms of their response to different polarizations of incident light, behavior in externally applied magnetic fields, and the anisotropic magnetic properties of Cu3Bi(SeO3)2O2Cl as determined by dc susceptibility measurements
High-vacuum-compatible high-power Faraday isolators for gravitational-wave interferometers
Faraday isolators play a key role in the operation of large-scale gravitational-wave detectors. Second-generation gravitational-wave interferometers such as the Advanced Laser Interferometer Gravitational-Wave Observatory (LIGO) and Advanced Virgo will use high-average-power cw lasers (up to 200 W) requiring specially designed Faraday isolators that are immune to the effects resulting from the laser beam absorptionâdegraded isolation ratio, thermal lensing, and thermally induced beam steering. In this paper, we present a comprehensive study of Faraday isolators designed specifically for high-performance operation in high-power gravitational-wave interferometers
Surgical site infection: poor compliance with guidelines and care bundles
Surgical site infections (SSIs) are probably the most preventable of the health care- associated infections. Despite the widespread international introduction of level I evidence-based guidelines for the prevention of SSIs, such as that of the National Institute for Clinical Excellence (NICE) in the UK and the surgical care improvement project (SCIP) of the USA, SSI rates have not measurably fallen. The care bundle approach is an accepted method of packaging best, evidence-based measures into routine care for all patients and, common to many guidelines for the prevention of SSI, includes methods for preoperative removal of hair (where appropriate), rational antibiotic prophylaxis, avoidance of perioperative hypothermia, management of perioperative blood glucose and effective skin preparation. Reasons for poor compliance with care bundles are not clear and have not matched the wide uptake and perceived benefit of the WHO âSafe Surgery Saves Livesâ checklist. Recommendations include the need for further research and continuous updating of guidelines; comprehensive surveillance, using validated definitions that facilitate benchmarking of anonymised surgeon-specific SSI rates; assurance that incorporation of checklists and care bundles has taken place; the development of effective communication strategies for all health care providers and those who commission services and comprehensive information for patients
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A specific amino acid motif of HLA-DRB1 mediates risk and interacts with smoking history in Parkinson's disease.
Parkinson's disease (PD) is a neurodegenerative disease in which genetic risk has been mapped to HLA, but precise allelic associations have been difficult to infer due to limitations in genotyping methodology. Mapping PD risk at highest possible resolution, we performed sequencing of 11 HLA genes in 1,597 PD cases and 1,606 controls. We found that susceptibility to PD can be explained by a specific combination of amino acids at positions 70-74 on the HLA-DRB1 molecule. Previously identified as the primary risk factor in rheumatoid arthritis and referred to as the "shared epitope" (SE), the residues Q/R-K/R-R-A-A at positions 70-74 in combination with valine at position 11 (11-V) is highly protective in PD, while risk is attributable to the identical epitope in the absence of 11-V. Notably, these effects are modified by history of cigarette smoking, with a strong protective effect mediated by a positive history of smoking in combination with the SE and 11-V (P = 10-4; odds ratio, 0.51; 95% confidence interval, 0.36-0.72) and risk attributable to never smoking in combination with the SE without 11-V (P = 0.01; odds ratio, 1.51; 95% confidence interval, 1.08-2.12). The association of specific combinations of amino acids that participate in critical peptide-binding pockets of the HLA class II molecule implicates antigen presentation in PD pathogenesis and provides further support for genetic control of neuroinflammation in disease. The interaction of HLA-DRB1 with smoking history in disease predisposition, along with predicted patterns of peptide binding to HLA, provide a molecular model that explains the unique epidemiology of smoking in PD
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