A Novel C-Terminal Homologue of Aha1 Co-Chaperone Binds to Heat Shock Protein 90 and Stimulates Its ATPase Activity in Entamoeba histolytica

Cytosolic heat shock protein 90 (Hsp90) has been shown to be essential for many infectious pathogens and is considered a potential target for drug development. In this study, we have carried out biochemical characterization of Hsp90 from a poorly studied protozoan parasite of clinical importance, Entamoeba histolytica. We have shown that Entamoeba Hsp90 can bind to both ATP and its pharmacological inhibitor, 17-AAG (17-allylamino-17-demethoxygeldanamycin), with K-d values of 365.2 and 10.77 mu M, respectively, and it has a weak ATPase activity with a catalytic efficiency of 4.12 x 10(-4) min(-1) mu M-1. Using inhibitor 17-AAG, we have shown dependence of Entamoeba on Hsp90 for its growth and survival. Hsp90 function is regulated by various co-chaperones. Previous studies suggest a lack of several important co-chaperones in E. histolytica. In this study, we describe the presence of a novel homologue of co-chaperone Aha1 (activator of Hsp90 ATPase), EhAha1c, lacking a canonical Aha1 N-terminal domain. We also show that EhAha1c is capable of binding and stimulating ATPase activity of EhHsp90. In addition to highlighting the potential of Hsp90 inhibitors as drugs against amoebiasis, our study highlights the importance of E. histolytica in understanding the evolution of Hsp90 and its co-chaperone repertoire. (C) 2014 Elsevier Ltd. All rights reserved.

Identification of heat shock factor binding protein in Plasmodium falciparum

Background: Heat shock factor binding protein (HSBP) was originally discovered in a yeast two-hybrid screen as an interacting partner of heat shock factor (HSF). It appears to be conserved in all eukaryotes studied so far, with yeast being the only exception. Cell biological analysis of HSBP in mammals suggests its role as a negative regulator of heat shock response as it appears to interact with HSF only during the recovery phase following exposure to heat stress. While the identification of HSF in the malaria parasite is still eluding biologists, this study for the first time, reports the presence of a homologue of HSBP in Plasmodium falciparum. Methods: PfHSBP was cloned and purified as his-tag fusion protein. CD (Circular dichroism) spectroscopy was performed to predict the secondary structure. Immunoblots and immunofluorescence approaches were used to study expression and localization of HSBP in P. falciparum. Cellular fractionation was performed to examine subcellular distribution of PfHSBP. Immunoprecipitation was carried out to identify HSBP interacting partner in P. falciparum. Results: PfHSBP is a conserved protein with a high helical content and has a propensity to form homo-oligomers. PfHSBP was cloned, expressed and purified. The in vivo protein expression profile shows maximal expression in trophozoites. The protein was found to exist in oligomeric form as trimer and hexamer. PfHSBP is predominantly localized in the parasite cytosol, however, upon heat shock, it translocates to the nucleus. This study also reports the interaction of PfHSBP with PfHSP70-1 in the cytoplasm of the parasite. Conclusions: This study emphasizes the structural and biochemical conservation of PfHSBP with its mammalian counterpart and highlights its potential role in regulation of heat shock response in the malaria parasite. Analysis of HSBP may be an important step towards identification of the transcription factor regulating the heat shock response in P. falciparum.

Two-step Synthesis of MoS2 Nanotubes using Shock Waves with Lead as Growth Promoter

A new two-step procedure for the synthesis of MoS2 nanotubes using lead as a growth promoter is reported. In the first step, molybdenum suboxide nanowhiskers containing a small amount of lead atoms were created by exposing a pressed MoS2+Pb mixture to highly compressed shock-heated argon gas, with estimated temperatures exceeding 9900 K. In the second step, these molybdenum suboxide nanowhiskers served as templates for the sulfidization of the oxide into MoS2 nanotubes (by using H2S gas in a reducing atmosphere at 820 degrees C). Unlike the case of WS2 nanotubes, the synthesis of a pure phase of MoS2 nanotubes from molybdenum oxide has proven challenging, due mostly to the volatile nature of the latter at the high requisite reaction temperatures (>800 degrees C). In contrast, the nature and apparent reaction mechanism of the method reported herein are amenable to future scale-up. The high-temperature shockwave system should also facilitate the synthesis of new nanostructures from other layered materials.

Stability of a circular tunnel in presence of pseudostatic seismic body forces

The stability of a long circular tunnel in a cohesive frictional soil medium has been determined in the presence of horizontal pseudo-static seismic body forces. The tunnel is supported by means of lining and anchorage system which is assumed to exert uniform internal compressive normal pressure on its periphery. The upper bound finite element limit analysis has been performed to compute the magnitude of the internal compressive pressure required to support the tunnel. The results have been presented in terms of normalized compressive normal stress, defined in terms of sigma(i)/c; where sigma(i) is the magnitude of the compressive normal pressure on the periphery of the tunnel and c refers to soil cohesion. The variation of sigma(i)/c with horizontal earthquake acceleration coefficient (alpha(h)) has been established for different combinations of H/D, gamma D/c and phi where (i) H and D refers to tunnel cover and diameter, respectively, and (ii) gamma and phi correspond to unit weight and internal friction angle of soil mass, respectively. Nodal velocity patterns have also been plotted for assessing the zones of significant plastic deformation. The analysis clearly reveals that an increase in the magnitude of the earthquake acceleration leads to a significant increment in the magnitude of internal compressive pressure. (C) 2014 Elsevier Ltd. All rights reserved.

Trans-spliced Heat Shock Protein 90 Modulates Encystation in Giardia lamblia

Background: Hsp90 from Giardia lamblia is expressed by splicing of two independently transcribed RNA molecules, coded by genes named HspN and HspC located 777 kb apart. The reasons underlying such unique trans-splicing based generation of GlHsp90 remain unclear. Principle Finding: In this study using mass-spectrometry we identify the sequence of the unique, junctional peptide contributed by the 5' UTR of HspC ORF. This peptide is critical for the catalytic function of Hsp90 as it harbours an essential ``Arg'' in its sequence. We also show that full length GlHsp90 possesses all the functional hall marks of a canonical Hsp90 including its ability to bind and hydrolyze ATP. Using qRT-PCR as well as western blotting approach we find the reconstructed Hsp90 to be induced in response to heat shock. On the contrary we find GlHsp90 to be down regulated during transition from proliferative trophozoites to environmentally resistant cysts. This down regulation of GlHsp90 appears to be mechanistically linked to the encystation process as we find pharmacological inhibition of GlHsp90 function to specifically induce encystation. Significance: Our results implicate the trans-spliced GlHsp90 from Giardia lamblia to regulate an essential stage transition in the life cycle of this important human parasite.

Heat shock protein 90 as a potential drug target against surra

Trypanosomiasis is caused by Trypanosoma species which affect both human and animal populations and pose a major threat to developing countries. The incidence of animal trypanosomiasis is on the rise. Surra is a type of animal trypanosomiasis, caused by Trypanosoma evansi, and has been included in priority list B of significant diseases by the World Organization of Animal Health (OIE). Control of surra has been a challenge due to the lack of effective drugs and vaccines and emergence of resistance towards existing drugs. Our laboratory has previously implicated Heat shock protein 90 (Hsp90) from protozoan parasites as a potential drug target and successfully demonstrated efficacy of an Hsp90 inhibitor in cell culture as well as a pre-clinical mouse model of trypanosomiasis. This article explores the role of Hsp90 in the Trypanosoma life cycle and its potential as a drug target. It appears plausible that the repertoire of Hsp90 inhibitors available in academia and industry may have value for treatment of surra and other animal trypanosomiasis.

Thermal Decomposition of Propargyl Alcohol: Single Pulse Shock Tube Experimental and ab Initio Theoretical Study

Thermal decomposition of propargyl alcohol (C3H3OH), a molecule of interest in interstellar chemistry and combustion, was investigated using a single pulse shock tube in the temperature ranging from 953 to 1262 K. The products identified include acetylene, propyne, vinylacetylene, propynal, propenal, and benzene. The experimentally observed overall rate constant for thermal decomposition of propargyl alcohol was found to be k = 10((10.17 +/- 0.36)) exp(-39.70 +/- 1.83)/RT) s(-1) Ab initio theoretical calculations were carried out to understand the potential energy surfaces involved in the primary and secondary steps of propargyl alcohol thermal decomposition. Transition state theory was used to predict the rate constants, which were then used and refined in a kinetic simulation of the product profile. The first step in the decomposition is C-O bond dissociation, leading to the formation of two important radicals in combustion, OH and propargyl. This has been used to study the reverse OH propargyl radical reaction, about which there appears to be no prior work. Depending on the site of attack, this reaction leads to propargyl alcohol or propenal, one of the major products at temperatures below 1200 K. A detailed mechanism has been derived to explain all the observed products.

Evaluating shock absorption behavior of small-sized systems under programmable electric field

A simple ball-drop impact tester is developed for studying the dynamic response of hierarchical, complex, small-sized systems and materials. The developed algorithm and set-up have provisions for applying programmable potential difference along the height of a test specimen during an impact loading; this enables us to conduct experiments on various materials and smart structures whose mechanical behavior is sensitive to electric field. The software-hardware system allows not only acquisition of dynamic force-time data at very fast sampling rate (up to 2 x 10(6) samples/s), but also application of a pre-set potential difference (up to +/- 10 V) across a test specimen for a duration determined by feedback from the force-time data. We illustrate the functioning of the set-up by studying the effect of electric field on the energy absorption capability of carbon nanotube foams of 5 x 5 x 1.2 mm(3) size under impact conditions. (C) 2014 AIP Publishing LLC.

Riemann shock tube: 1D normal shocks in air, simulations and experiments

This paper presents numerical simulation of the evolution of one-dimensional normal shocks, their propagation, reflection and interaction in air using a single diaphragm Riemann shock tube and validate them using experimental results. Mathematical model is derived for one-dimensional compressible flow of viscous and conducting medium. Dimensionless form of the mathematical model is used to construct space-time finite element processes based on minimization of the space-time residual functional. The space-time local approximation functions for space-time p-version hierarchical finite elements are considered in higher order GRAPHICS] spaces that permit desired order of global differentiability of local approximations in space and time. The resulting algebraic systems from this approach yield unconditionally positive-definite coefficient matrices, hence ensure unique numerical solution. The evolution is computed for a space-time strip corresponding to a time increment Delta t and then time march to obtain the evolution up to any desired value of time. Numerical studies are designed using recently invented hand-driven shock tube (Reddy tube) parameters, high/low side density and pressure values, high- and low-pressure side shock tube lengths, so that numerically computed results can be compared with actual experimental measurements.

Anthrapyrazolone analogues intercept inflammatory JNK signals to moderate endotoxin induced septic shock

Severe sepsis or septic shock is one of the rising causes for mortality worldwide representing nearly 10% of intensive care unit admissions. Susceptibility to sepsis is identified to be mediated by innate pattern recognition receptors and responsive signaling pathways of the host. The c-Jun N-terminal Kinase (JNK)-mediated signaling events play critical role in bacterial infection triggered multi-organ failure, cardiac dysfunction and mortality. In the context of kinase specificities, an extensive library of anthrapyrazolone analogues has been investigated for the selective inhibition of c-JNK and thereby to gain control over the inflammation associated risks. In our comprehensive biochemical characterization, it is observed that alkyl and halogen substitution on the periphery of anthrapyrazolone increases the binding potency of the inhibitors specifically towards JNK. Further, it is demonstrated that hydrophobic and hydrophilic interactions generated by these small molecules effectively block endotoxin-induced inflammatory genes expression in in vitro and septic shock in vivo, in a mouse model, with remarkable efficacies. Altogether, the obtained results rationalize the significance of the diversity oriented synthesis of small molecules for selective inhibition of JNK and their potential in the treatment of severe sepsis.

Study the effect of electrical and mechanical shock loading on liberation and milling characteristics of mineral materials

Milling is an energy intensive process and it is considered as one of the most energy inefficient processes. Electrical and mechanical shock loading can be used to develop a pre-treatment methodology to enhance energy efficiency of comminution and liberation of minerals. Coal and Banded Hematite Jasper (BHJ) Iron ores samples were taken for the study to know the effect of shock loading. These samples were exposed to 5 electric shocks of 300 kV using an electric shock loading device. A diaphragmless shock tube was used to produce 3 and 6 compressed air shocks of Mach number 2.12 to treat the coal and Iron ore samples. Microscopic, comminution and liberation studies were carried out to compare the effectiveness of these approaches. It was found that electric shock loading can comminute the coal samples more effectively and increases the yield of carbon by 40% at 1.6 gm/cc density over the untreated coal samples. Mechanical shock loading showed improved milling performance for both the materials and 12.90% and 8.1% reduction in the D-80 of the particles was observed during grinding for treated samples of coal and iron, respectively. Liberation of minerals in BHJ Iron ore was found unaffected due to low intensity of the mechanical shock waves and non conductivity of minerals. Compressed air based shock loading is easier to operate than electrical shock loading and it needs to be explored further to improve the energy efficacy of comminution. (C) 2014 Elsevier Ltd. All rights reserved.

Large carbon cluster thin film gauges for measuring aerodynamic heat transfer rates in hypersonic shock tunnels

Different types of Large Carbon Cluster (LCC) layers are synthesized by a single-step pyrolysis technique at various ratios of precursor mixture. The aim is to develop a fast responsive and stable thermal gauge based on a LCC layer which has relatively good electrical conduction in order to use it in the hypersonic flow field. The thermoelectric property of the LCC layer has been studied. It is found that these carbon clusters are sensitive to temperature changes. Therefore suitable thermal gauges were developed for blunt cone bodies and were tested in hypersonic shock tunnels at a flow Mach number of 6.8 to measure aerodynamic heating. The LCC layer of this thermal gauge encounters high shear forces and a hostile environment for test duration in the range of a millisecond. The results are favorable to use large carbon clusters as a better sensor than a conventional platinum thin film gauge in view of fast responsiveness and stability.

Response of shock wave deformation in AA5086 aluminum alloy

The AA5086 aluminum alloy sheets with different starting textures were subjected to shock wave deformation with an input impulse of similar to 0.2 Ns. Microstructural examination indicate no significant change in grain size; however, the evolution of substructure manifesting intra-granular misorientation was evident. The improvement in hardness indicates the absence of recovery and strain hardening during shock deformation. Shock deformed samples show characteristic texture evolution with high Brass {110}< 112 > component. The study demonstrates the viability of high velocity forming of AA5086 aluminum alloy sheet using shock wave. (C) 2014 Elsevier B.V. All rights reserved.

Remotely triggered micro-shock wave responsive drug delivery system for resolving diabetic wound infection and controlling blood sugar levels

A novel, micro-shock wave responsive spermidine and dextran sulfate microparticle was developed. Almost 90% of the drug release was observed when the particles were exposed to micro-shock waves 5 times. Micro-shock waves served two purposes; of releasing the antibiotic from the system and perhaps disrupting the S. aureus biofilm in the skin infection model. A combination of shock waves with ciprofloxacin loaded microparticles could completely cure the S. aureus infection lesion in a diabetic mouse model. As a proof of concept insulin release was triggered using micro-shock waves in diabetic mice to reduce the blood glucose level. Insulin release could be triggered for at least 3 days by exposing subcutaneously injected insulin loaded particles.

Role of boron diffusion in CoFeB/MgO magnetic tunnel junctions

Several scientific issues concerning the latest generation read heads for magnetic storage devices, based on CoFeB/MgO/CoFeBmagnetic tunnel junctions (MTJs) are known to be controversial, including such fundamental questions as to the behavior and the role of B in optimizing the physical properties of these devices. Quantitatively establishing the internal structures of several such devices with different annealing conditions using hard x-ray photoelectron spectroscopy, we resolve these controversies and establish that the B diffusion is controlled by the capping Ta layer, though Ta is physically separated from the layer with B by several nanometers. While explaining this unusual phenomenon, we also provide insight into why the tunneling magnetoresistance (TMR) is optimized at an intermediate annealing temperature, relating it to B diffusion, coupled with our studies based on x-ray diffraction and magnetic studies.