The complementation of yeast with human or Plasmodium falciparum Hsp90 confers differential inhibitor sensitivities

Developing novel drugs against the unicellular parasite Plasmodium is complicated by the paucity of simple screening systems. Heat-shock proteins are an essential class of proteins for the parasite's cyclical life style between different cellular milieus and temperatures. The molecular chaperone Hsp90 assists a large variety of proteins, but its supporting functions for many proteins that are important for cancer have made it into a well-studied drug target. With a better understanding of the differences between Hsp90 of the malarial parasite and Hsp90 of its human host, new therapeutic options might become available. We have generated a set of isogenic strains of the budding yeast Saccharomyces cerevisiae where the essential yeast Hsp90 proteins have been replaced with either of the two human cytosolic isoforms Hsp90 alpha or Hsp90 beta, or with Hsp90 from Plasmodium falciparum (Pf). All strains express large amounts of the Flag-tagged Hsp90 proteins and are viable. Even though the strain with Pf Hsp90 grows more poorly, it provides a tool to reconstitute additional aspects of the parasite Hsp90 complex and its interactions with substrates in yeast as a living test tube. Upon exposure of the set of Hsp90 test strains to the two Hsp90 inhibitors radicicol (Rd) and geldanamycin (GA), we found that the strain with Pf Hsp90 is relatively more sensitive to GA than to Rd compared to the strains with human Hsp90's. This indicates that this set of yeast strains could be used to screen for new Pf Hsp90 inhibitors with a wider therapeutic window.

Modeling of seepage flow through layered solis

A two-dimensional finite difference model, which solves mixed type of Richards' equation, whose non-linearity is dealt with modified Picard's iteration and strongly implicit procedure to solve the resulting equations, is presented. Modeling of seepage flow through heterogeneous soils, which is common in the field is addressed in the present study. The present model can be applied to both unsaturated and saturated soils and can handle very dry initial condition and steep wetting fronts. The model is validated by comparing experimental results reported in the literature. Newness of this two dimensional model is its application on layered soils with transient seepage face development, which has not been reported in the literature. Application of the two dimensional model for studying unconfined drainage due to sudden drop of water table at seepage face in layered soils is demonstrated. In the present work different sizes of rectangular flow domain with different types of layering are chosen. Sensitivity of seepage height due to problem dimension of layered system is studied. The effect of aspect ratio on seepage face development in case of the flow through layered soil media is demonstrated. The model is also applied to random heterogeneous soils in which the randomness of the model parameters is generated using the turning band technique. The results are discussed in terms of phreatic surface and seepage height development and also flux across the seepage face. Such accurate modeling of seepage face development and quantification of flux moving across the seepage face becomes important while modeling transport problems in variably saturated media.