Detailed Structure–Function Correlations of Bacillus subtilis Acetolactate Synthase

Isobutanol is deemed to be a next‐generation biofuel and a renewable platform chemical. 1 Non‐natural biosynthetic pathways for isobutanol production have been implemented in cell‐based and in vitro systems with Bacillus subtilis acetolactate synthase (AlsS) as key biocatalyst. 2 – 6 AlsS catalyzes the condensation of two pyruvate molecules to acetolactate with thiamine diphosphate and Mg²⁺ as cofactors. AlsS also catalyzes the conversion of 2‐ketoisovalerate into isobutyraldehyde, the immediate precursor of isobutanol. Our phylogenetic analysis suggests that the ALS enzyme family forms a distinct subgroup of ThDP‐dependent enzymes. To unravel catalytically relevant structure‐function relationships, we solved the AlsS crystal structure at 2.3 Å in the presence of ThDP, Mg²⁺ and in a transition state with a 2‐lactyl moiety bound to ThDP. We supplemented our structural data by point mutations in the active site to identify catalytically important residues.     

Bake oven induced variation of surface chemistry on electrocoat paint: Effect on primer-electrocoat intercoat adhesion

The interplay of bake oven processing environment and surface/interfacial chemistry, and their effect on intercoat adhesion has been investigated for a model primer/electrocoat paint system. X-ray photoelectron spectroscopy (XPS) surface analysis established that a prominent ether component in the C 1s core level spectrum correlated with a polyether-based crater-control additive in the electrocoat that surface-segregates during cure. Laboratory-simulated bake oven experiments confirmed that better adhesion characteristics were realized by removal of this electrocoat overlayer through reaction with nitrogen oxides and high levels of moisture present in the cure environment of production direct-fired ovens. Details of the effect of bake oven atmosphere on cure chemistry for direct- versus indirect-fired ovens are presented.     

Escherichia coli and enterococci at beaches in the Grand Traverse Bay,Lake Michigan: sources,characteristics, and environmental pathways

This study quantified Escherichia coli (EC) and enterococci (ENT) in beach waters and dominant source materials, correlated these with ambient conditions, and determined selected EC genotypes and ENT phenotypes. Bathing-water ENT criteria were exceeded more frequently than EC criteria, providing conflicting interpretations of water quality. Dominant sources of EC and ENT were bird feces (10(8)/d/bird), storm drains (10(7)/d), and river water (10(11)/d); beach sands, shallow groundwater and detritus were additional sources. Beach-water EC genotypes and ENT phenotypes formed clusters with those from all source types, reflecting diffuse inputs. Some ENT isolates had phenotypes similar to those of human pathogens and/or exhibited high-level resistance to human-use antibiotics. EC and ENT concentrations were influenced by collection time and wind direction. There was a 48-72-h lag between rainfall and elevated EC concentrations at three southern shoreline beaches, but no such lag at western and eastern shoreline beaches, reflecting the influence of beach orientation with respect to cyclic (3-5 d) summer weather patterns. In addition to local contamination sources and processes, conceptual or predictive models of Great Lakes beach water quality should consider regional weather patterns, lake hydrodynamics, and the influence of monitoring method variables (time of day, frequency).     

An analysis of thematic mapper simulator data for urban environments

Thematic mapper simulator data collected for the Los Angeles Basin in 1980 were examined to assess their utility for urban and near-urban land-cover delimitations. Spectral data for six of the thematic mapper channels were reprojected to a UTM grid and aggregated to 30-m resolution, 120 m for the thermal band. Statistics for 21 training sites representing 8 land-cover types were obtained and examined using transformed divergence calculations for intraclass variability, optimal number of channels for classification, and best channels for classification. Four channels of data are adequate for classification with the best results obtained by selection of one channel from each of the available major portions of the electromagnetic spectrum. The thermal channel data is useful for urban land-cover delineations at 30-m resolution, but its utility at 120-m resolution is not clear from this study.     

Quantitative liver ADC measurements using diffusion-weighted MRI at 3 Tesla: evaluation of reproducibility and perfusion dependence using different techniques for respiratory compensation

Object

Diffusion weighted imaging (DWI) of the liver suffers from low signal to noise making 3 Tesla (3 T) an attractive option, but 3 T data is scarce. It was the aim to study the influence of different b values and respiratory compensation methods (RCM) on the apparent diffusion coefficient (ADC) level and on ADC reproducibility at 3 T.

Materials and methods

Ten healthy volunteers and 12 patients with malignant liver lesions underwent repeated (2–22 days) breathhold, free-breathing and respiratory triggered DWI at 3 T using b values between 0 and 1,000 s/mm².

Results

The ADCs changed up to 150 % in healthy livers and up to 48 % in malignant lesions depending on b value combinations. Best ADC reproducibility in healthy livers were obtained with respiratory triggering (95 % limits of agreement: ±0.12) and free-breathing (±0.14). In malignant lesions equivalent reproducibility was obtained with less RCM dependence. The use of a lower maximum b value (b = 500) decreased reproducibility (±0.14 to ±0.32) in both normal liver and malignant lesions.

Conclusion

Large differences in absolute ADC values and reproducibility caused by varying combinations of clinically realistic b values were demonstrated. Different RCMs caused smaller differences. Lowering maximum b value to 500 increased limits of agreement up to a factor of two. Serial ADC changes larger than approximately 15 % can be detected confidently on an individual basis in both malignant lesions and normal liver parenchyma at 3 T using appropriate b values and respiratory compensation.     

An analysis of stresses in a model of the periodontal ligament

The writers analyze the stresses in a model of the periodontal ligament of a maxillary central incisor. The model is assumed to be of paraboloidal shape, uniform thickness, and of a homogeneous, isotropic, and linearly elastic material. The inside surface of this paraboloidal shell is subject to rigid body displacement while the outside surface is assumed rigidly supported and stationary. The edge of the model is assumed to be stress free.

A system of paraboloidal coordinates is developed, and this system is fitted to the shape of the root of a typical maxillary central incisor. The displacement functions obtained by solving the differential equations of equilibrium are used to determine the stresses at the inside and outside surfaces of the model ligament. These stresses are subject to superposition so that the stresses associated with any combination of applied forces and couples may be determined.     

Fire protection in traffic tunnels: General aspects and results of the EUREKA project

Against the background of increasing problems concerning safety in underground transport, facilities, nine European nations cooperated in the EUREKA project EU 499 FIRETUN. Their common efforts formed the basis for a remarkable condensed instrumentation of a 2300-m-long test tunnel. Altogether, 20 tests were conducted between 1990 and 1992 and evaluated from 1993 through 1995. The many important results gained during the project form a strong basis for international discussion of fire safety concepts.