Optical spectroscopy of nicotinoprotein alcohol dehydrogenase from Amycolatopsis methanolica: a comparison with horse liver alcohol dehydrogenase and UDP-galactose epimerase

The NADH absorbance spectrum of nicotinoprotein (NADH-containing) alcohol dehydrogenase from Amycolatopsis methanolica has a maximum at 326 nm. Reduced enzyme-bound pyridine dinucleotide could be reversibly oxidized by acetaldehyde. The fluorescence excitation spectrum for NADH bound to the enzyme has a maximum at 325 nm. Upon excitation at 290 nm, energy transfer from tryptophan to enzyme-bound NADH was negligible. The fluorescence emission spectrum (excitation at 325 nm) for NADH bound to the enzyme has a maximum at 422 nm. The fluorescence intensity is enhanced by a factor of 3 upon binding of isobutyramide (Kd = 59 microM). Isobutyramide acts as competitive inhibitor (Ki = 46 microM) with respect to the electron acceptor NDMA (N,N-dimethyl-p-nitrosoaniline), which binds to the enzyme containing the reduced cofactor. The nonreactive substrate analogue trifluoroethanol acts as a competitive inhibitor with respect to the substrate ethanol (Ki = 1.6 microM), which binds to the enzyme containing the oxidized cofactor. Far-UV circular dichroism spectra of the enzyme containing NADH and the enzyme containing NAD+ were identical, indicating that no major conformational changes occur upon oxidation or reduction of the cofactor. Near-UV circular dichroism spectra of NADH bound to the enzyme have a minimum at 323 nm (Deltaepsilon = -8.6 M-1 cm-1). The fluorescence anisotropy decay of enzyme-bound NADH showed no rotational freedom of the NADH cofactor. This implies a rigid environment as well as lack of motion of the fluorophore. The average fluorescence lifetime of NADH bound to the enzyme is 0.29 ns at 20 degreesC and could be resolved into at least three components (in the range 0.13-0.96 ns). Upon binding of isobutyramide to the enzyme-containing NADH, the average excited-state lifetime increased to 1.02 ns and could be resolved into two components (0.37 and 1.11 ns). The optical spectra of NADH bound to nicotinoprotein alcohol dehydrogenase have blue-shifted maxima compared to other NADH-dehydrogenase complexes, but comparable to that observed for NADH bound to horse liver alcohol dehydrogenase. The fluorescence lifetime of NADH bound to the nicotinoprotein is very short compared to enzyme-bound NADH complexes, also compared to NADH bound to horse liver alcohol dehydrogenase. The cofactor-protein interaction in the nicotinoprotein alcohol dehydrogenase active site is more rigid and apolar than that in horse liver alcohol dehydrogenase. The optical properties of NADH bound to nicotinoprotein alcohol dehydrogenase differ considerably from NADH (tightly) bound to UDP-galactose epimerase from Escherichia coli. This indicates that although both enzymes have NAD(H) as nonexchangeable cofactor, the NADH binding sites are quite different.     

Three-dimensional respiratory-gated MR angiography of coronary arteries: comparison with conventional coronary angiography

OBJECTIVE: MR coronary angiography is most often performed using two-dimensional techniques. Although three-dimensional (3D) acquisitions do have important advantages, they take too long for a single breath-hold and are thus susceptible to respiratory motion artifacts. The purpose of this study was to investigate the accuracy of a unique respiratory-gated 3D MR angiographic technique in identifying the proximal coronary arteries in patients suspected of having coronary artery disease. In addition, we investigated the capability of this technique to detect proximal stenoses. SUBJECTS AND METHODS: We performed a prospective blinded study in 20 patients who were referred for conventional coronary angiography. A cardiac-gated 3D gradient-echo sequence with fat suppression was used. Retrospective respiratory gating was performed using navigator echoes of the diaphragm position. Using multiplanar reformatting, two independent readers blindly analyzed the data sets for visualization of major coronary arteries, lengths of imaged segments, and detection of significant stenoses (> 50% occlusion of the luminal diameter by conventional angiography). RESULTS: Seventy-seven of 80 (96%) coronary arteries were positively identified. In one patient, an anomalous coronary anatomy was readily identified and confirmed by conventional angiography. The average lengths of the imaged segments of the right, left main, left anterior descending, and left circumflex coronary arteries were 58 +/- 13 mm, 9 +/- 5 mm, 59 +/- 16 mm, and 24 +/- 10 mm, respectively. Overall sensitivity for the detection of stenoses was low (38%), with a specificity of 95%. Interobserver agreement was 0.92, with a kappa value of 0.65. CONCLUSION: Respiratory-gated 3D MR angiography allows accurate identification of proximal coronary arteries and may be valuable for 3D imaging of coronary anomalies. Further technical improvements are required to enhance the value of the technique in detecting stenoses.     

A System for the Rapid Acquisition of Surface Potential Maps of Human Skeletal Muscle Motor Units

Inability to predict the intramusclar orientation, depth, and distribution of a motor unit's muscle fibers within a human skeletal muscle complicates unit size estimation. A data collection system is described that is suitable for systematic study of this problem.     

Instructions for Authors

Instructions for Authors

Instructions for Authors     

Attenuated total reflection design for in situ FT-IR spectroelectrochemical studies

A versatile spectroelectrochemical apparatus is introduced to study the changes in IR spectra of organic and inorganic compounds upon oxidation or reduction. The design is based on an attenuated total reflection device, which permits the study of a wide spectral range of 16,700 (600 nm)-250 cm(-1), with a small opaque region of 2250-1900 cm(-1). In addition, an IR data collection protocol is introduced to deal with electrochemically nonreversible background signals. This method is tested with ferrocene in acetonitrile; concentrations as low as 1 mM produce results that agree with those in the literature.     

Experimental study of a shearing interferometer concept for at-wavelength characterization of extreme-ultraviolet optics

We describe the experimental evaluation of a shearing interferometer concept for at-wavelength testing of extreme-ultraviolet optics. The concept is based on the Ronchi test, which has been modified by a new design for entrance and exit gratings to suppress disturbing higher-order interference patterns. The interferometer concept has been tested on an experimental setup, of which all relevant parameters have been scaled from extreme-ultraviolet to visible-light wavelengths. A Twyman-Green interferometer has been integrated into the setup for comparison with the improved Ronchi test. A systematic difference of 7-12 mlambda rms has been found between wave fronts measured with the improved Ronchi test and with the Twyman-Green interferometer. Possible error sources have been analyzed. The accuracy of the interferometer is estimated to be 10 mlambda rms.     

A multicomponential model of authenticity.

A multicomponential model of authenticity is presented which includes psychosocial, cultural, intrapsychic, personality and capacity related and neurobiological aspects of authenticity. Genetic, political and ethnic influences could also involved in authentic etiology. More research is needed into the correlates (and their interactions) of authenticity in order to develop adequate intervention and prevention programs for individuals who demonstrate a lack of authenticity. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Cognitive and serotonergic vulnerability to depression: Convergent findings.

Cognitive reactivity (CR) is a psychological vulnerability marker of depression, whereas response to acute tryptophan depletion (ATD; a serotonergic challenge procedure) is a biological vulnerability marker. The aim of this study was to investigate the relationship between these markers. Thirty-nine remitted depressed patients participated in 2 ATD sessions in a double-blind crossover design. CR, assessed prior to the ATD sessions, predicted depressive response to high-dose ATD. CR also diminished the effects of 2 known predictors of ATD response: gender and residual symptoms. Neuroticism and behavioral inhibition were unrelated to ATD response. CR is associated with an increased sensitivity to reductions of serotonin concentrations. These findings present a small step toward unifying cognitive and neurobiological theories of depression. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Ring-opening Metathesis Co-polymerization of a C60-cyclopentadiene Cycloadduct and Norbornene with the Grubbs Second-generation Catalyst

A series of C₆₀-containing polymers were synthesized by the co-polymerization of a C₆₀-cyclopentadiene cycloadduct and norbornene in varying ratios. The polymerization was facilitated by a catalytic amount of Grubbs second generation catalyst and the co-polymers formed were investigated by spectroscopic and thermal techniques.     

Robust dimensions of anxiety sensitivity: Development and initial validation of the Anxiety Sensitivity Index-3.

Accumulating evidence suggests that anxiety sensitivity (fear of arousal-related sensations) plays an important role in many clinical conditions, particularly anxiety disorders. Research has increasingly focused on how the basic dimensions of anxiety sensitivity are related to various forms of psychopathology. Such work has been hampered because the original measure--the Anxiety Sensitivity Index (ASI)--was not designed to be multidimensional. Subsequently developed multidimensional measures have unstable factor structures or measure only a subset of the most widely replicated factors. Therefore, the authors developed, via factor analysis of responses from U.S. and Canadian nonclinical participants (n = 2,361), an 18-item measure, the ASI-3, which assesses the 3 factors best replicated in previous research: Physical, Cognitive, and Social Concerns. Factorial validity of the ASI-3 was supported by confirmatory factor analyses of 6 replication samples, including nonclinical samples from the United States and Canada, France, Mexico, the Netherlands, and Spain (n = 4,494) and a clinical sample from the United States and Canada (n = 390). The ASI-3 displayed generally good performance on other indices of reliability and validity, along with evidence of improved psychometric properties over the original ASI. (PsycINFO Database Record (c) 2010 APA, all rights reserved)