Characterization of a site-directed mutant of cytochrome b5 designed to alter axial imidazole ligand plane orientation

Mutants of cytochrome b5 were designed to achieve reorientation of individual axial imidazole ligands. The orientation of the axial ligand planes is thought to modulate the reduction potential of bis(imidazole) axially ligated heme proteins. The A67V mutation achieved this goal through the substitution of a bulkier, hydrophobic ligand for a residue, in the sterically hindered hydrophobic heme binding pocket. Solution structures of mutant and wild-type proteins in the region of the mutation were calculated using restraints obtained from 1H and 15N 2D homonuclear and heteronuclear NMR spectra and 1H-15N 3D heteronuclear NMR spectra. More than 10 restraints per residue were used in the refinement of both structures. Average local rmsd for 20 refined structures was 0.30 A for the wild-type structure and 0.38 A for the A67V mutant. The transfer of amide proton resonance assignments from wild-type to the mutant protein was achieved through overlays of 15N-1H heteronuclear correlation spectra of the reduced proteins. Side chain assignments and sequential assignments were established using conventional assignment strategies. Calculation of the orientation of the components of the anisotropic paramagnetic susceptibility tensor, using methods similar to procedures applied to the wild-type protein, shows that the orientation of the in-plane components are identical in the wild-type and mutant proteins. However, the orientation of the z-component of the susceptibility tensor calculated for the mutant protein differs by 17 degrees for the A-form and by 11 degrees for the B-form from the orientation calculated for the wild-type protein. The rotation of the z-component of the susceptibility tensor (toward the delta meso proton) is in the same direction and is of the same magnitude as the rotation of the H63 imidazole ring induced by mutation.     

Pseudoactive bridge for separation of R and C in a practical capacitive/conductive transducer

An effective and unique method of measuring independently the variations of resistive and capacitive constituents of a practical capacitive transducer using an improved pseudo-active bridge is described. The impedance change in some types of transducers may be due to changes in resistance and/or capacitance either way. Separation of changes in resistance and capacitance results in a better understanding of the transduced variable. The active bridge which separates the changes is suitable for online determination of the two changes.     

Influence of long- term aging and superimposed creep

In an effort to understand the effects of long-term aging at high temperatures and those of the superimposed creep stresses on the microstructural variations in a 0.50Cr-0.50Mo-0.25V steel, the shoulder and gage portions of the specimens subjected to stress rupture tests at 500 °C and 540 °C have been studied by transmission electron microscopy. The optical microstructure, in the normalized and tempered condition, consists of about 95 pct ferrite with the remainder bainite. The ferrite has an extremely fine precipitation of vanadium carbide (VC) as interphase precipitate as well as in random distribution. In the bainitic area, M₃C is present in the globular as well as plate-like morphology. Specimens ruptured at 500 °C and 540 °C were selected for transmission electron microscopic study. Both the interphase precipitates and randomly distrib- uted precipitates of VC coarsen considerably due to prolonged exposure at elevated tempera- tures, and ultimately fine M₂C platelets nucleate and grow on either side of these precipitates, resulting in the formation of the so-called H-carbides. The superimposed creep stress has a marginal effect on the coarsening kinetics of VC but has enhanced its precipitation along dis- locations and has promoted the formation of H-carbides.     

Estimation of fuzzy memberships from histograms

Based on the conclusions drawn in the bijective transformation between possibility and probability, a method is proposed to estimate the fuzzy membership function for pattern recognition purposes. A rational function approximation to the probability density function is obtained from the histogram of a finite (and sometimes very small) number of samples. This function is normalized such that the highest ordinate is one. The parameters representing the rational function are used for classifying the pattern samples based on a max-min decision rule. The method is illustrated with examples.     

Water resources utilization in an irrigation project in India

Quantitative information on the adequacy of current operating procedures and available water resources in a typical major irrigation project in India (the Mahi Right Bank Canal project in Gujarat, India) is provided by comparing monthly and seasonal irrigation demands with canal releases and groundwater draught over a four‐year period. A strategy based on water conservation and the use of groundwater in the monsoon season for the full exploitation of reservoir potential in the subsequent seasons is proposed.     

An intermittently used n-unit complex system

In this paper we study an n-unit warm standby system which is needed intermittently with need and no-need periods occurring alternately. The life time of a unit while online and duration of a no-need period are generally distributed random variables while all the others are exponentially distributed. Identifying certain regeneration points, we obtain an expression for the distribution function of the time to the first disappointment.