Type IA topoisomerase inhibition by clamp closure

Bacterial DNA topoisomerase I (topoI) catalyzes relaxation of negatively supercoiled DNA. The enzyme alters DNA topology through protein-operated DNA gate, switching between open and closed conformations during its reaction. We describe the mechanism of inhibition of Mycobacterium smegmatis and Mycobacterium tuberculosis topoI by monoclonal antibodies (mAbs) that bind with high affinity and inhibit at 10-50 nM concentration. Unlike other inhibitors of topoisomerases, the mAbs inhibited several steps of relaxation reaction, namely DNA binding, cleavage, strand passage, and enzyme-DNA dissociation. The enhanced religation of the cleaved DNA in presence of the mAb indicated closing of the enzyme DNA gate. The formation of enzyme-DNA heterocatenane in the presence of the mAbs as a result of closing the gate could be inferred by the salt resistance of the complex, visualized by atomic force microscopy and confirmed by fluorescence measurements. Locking the enzyme-DNA complex as a closed clamp restricted the movements of the DNA gate, affecting all of the major steps of the relaxation reaction. Enzyme trapped on DNA in closed clamp conformation formed roadblock for the elongating DNA polymerase. The unusual multistep inhibition of mycobacterial topoisomerases may facilitate lead molecule development, and the mAbs would also serve as valuable tools to probe the enzyme mechanism.

Curvature based mobility analysis and form closure of smooth planar curves with multiple contacts

This paper presents a simple second-order, curvature based mobility analysis of planar curves in contact. The underlying theory deals with penetration and separation of curves with multiple contacts, based on relative configuration of osculating circles at points of contact for a second-order rotation about each point of the plane. Geometric and analytical treatment of mobility analysis is presented for generic as well as special contact geometries. For objects with a single contact, partitioning of the plane into four types of mobility regions has been shown. Using point based composition operations based on dual-number matrices, analysis has been extended to computationally handle multiple contacts scenario. A novel color coded directed line has been proposed to capture the contact scenario. Multiple contacts mobility is obtained through intersection of the mobility half-spaces. It is derived that mobility region comprises a pair of unbounded or a single bounded convex polygon. The theory has been used for analysis and synthesis of form closure configurations, revolute and prismatic kinematic pairs. (C) 2013 Elsevier Ltd. All rights reserved.

Combining data sets of satellite-retrieved products for basin-scale water balance study: 2. Evaluation on the Mississippi Basin and closure correction model

In this study, we applied the integration methodology developed in the companion paper by Aires (2014) by using real satellite observations over the Mississippi Basin. The methodology provides basin-scale estimates of the four water budget components (precipitation P, evapotranspiration E, water storage change Delta S, and runoff R) in a two-step process: the Simple Weighting (SW) integration and a Postprocessing Filtering (PF) that imposes the water budget closure. A comparison with in situ observations of P and E demonstrated that PF improved the estimation of both components. A Closure Correction Model (CCM) has been derived from the integrated product (SW+PF) that allows to correct each observation data set independently, unlike the SW+PF method which requires simultaneous estimates of the four components. The CCM allows to standardize the various data sets for each component and highly decrease the budget residual (P - E - Delta S - R). As a direct application, the CCM was combined with the water budget equation to reconstruct missing values in any component. Results of a Monte Carlo experiment with synthetic gaps demonstrated the good performances of the method, except for the runoff data that has a variability of the same order of magnitude as the budget residual. Similarly, we proposed a reconstruction of Delta S between 1990 and 2002 where no Gravity Recovery and Climate Experiment data are available. Unlike most of the studies dealing with the water budget closure at the basin scale, only satellite observations and in situ runoff measurements are used. Consequently, the integrated data sets are model independent and can be used for model calibration or validation.

MOMENT ANALYSIS OF A SCROLL EXPANDER USED IN AN ORGANIC RANKINE CYCLE

Recent studies on small-scale power generation with the organic Rankine cycle suggest superior performance of positive displacement type of expanders compared to turbines. Scroll expanders in particular achieve high isentropic efficiencies due to lower leakage and frictional losses. Performance of scroll machines may be enhanced by the use of non-circular involute curves in place of the circular involutes resulting non-uniform wall thickness. In this paper, a detailed moment analysis is performed for such an expander having volumetric expansion ratio of 5 using thermodynamic models proposed earlier by one of the present authors. The working fluid considered in the power cycle is R-245fa with scroll inlet temperature of 125 degrees C for a gross power output of similar to 3.5 kW. The model developed in this paper is verified with an air scroll compressor available in the literature and then applied to an expander Prediction of small variation of moment with scroll motion recommends use of scroll expander without a flywheel over other positive displacement type of expanders, e.g. reciprocating, where a flywheel is an essential component.