Tracking the Interplay between Bound Peptide and the Lid Domain of DnaK,Using Molecular Dynamics

Hsp70 chaperones consist of two functional domains: the 44 kDa Nucleotide Binding Domain (NBD), that binds and hydrolyses ATP, and the 26 kDa Substrate Binding Domain (SBD), which binds unfolded proteins and reactivates them, utilizing energy obtained from nucleotide hydrolysis. The structure of the SBD of the bacterial Hsp70, DnaK, consists of two sub-domains: A β-sandwich part containing the hydrophobic cavity to which the hepta-peptide NRLLLTG (NR) is bound, and a segment made of 5 α-helices, called the “lid” that caps the top of the β-sandwich domain. In the present study we used the Escherichia coli Hsp70, DnaK, as a model for Hsp70 proteins, focusing on its SBD domain, examining the changes in the lid conformation. We deliberately decoupled the NBD from the SBD, limiting the study to the structure of the SBD section, with an emphasis on the interaction between the charges of the peptide with the residues located in the lid. Molecular dynamics simulations of the complex revealed significant mobility within the lid structure; as the structure was released from the forces operating during the crystallization process, the two terminal helices established a contact with the positive charge at the tip of the peptide. This contact is manifested only in the presence of electrostatic attraction. The observed internal motions within the lid provide a molecular role for the function of this sub-domain during the reaction cycle of Hsp 70 chaperones.     

Reversible,repeatable and low phase transition behaviour of spin coated nanostructured vanadium oxide thin films with superior mechanical properties

Smooth, uniform and crystalline vanadium oxide thin films were deposited on quartz by spin coating technique with four different rpm i.e., 1000, 2000, 3000 and 4000 and subsequently post annealed at 350, 450 and 550?°C in vacuum. Transmission electron microscopy (TEM), Field emission scanning electron microscopy (FESEM) and X-ray diffraction (XRD) techniques were utilized for microstructural characterizations and phase analysis, respectively, for vanadium oxide powder and deposited film. Nanorods were observed to be grown after vacuum annealing. X-ray photoelectron spectroscopy (XPS) technique was utilized to study the elemental oxidation state of deposited vanadium oxide films. Thermo-optical and electrical properties such as solar transmittance (τ_s), reflectance (ρ_s), absorptance (α_s), infrared (IR) emittance (ε_ir) and sheet resistance (R_s) of different thin films were evaluated. Based on the optical characteristics the optimized condition of the film processing was identified to be spin coated at 3000?rpm. Subsequently, the nanoindentation technique was utilized to measure hardness and Young's modulus of the optimized film. The measured nanomechanical properties were found to be superior to those reported for sputtered vanadium oxide films. Finally, temperature dependent phase transition characteristics of optimized vanadium oxide films were studied by differential scanning calorimetry (DSC) technique. Reversible and repeatable phase transition was found to occur in the range of 44–48?°C which was significantly lower than the phase transition temperature (i.e., 68?°C) of bulk VO₂.     

Gauging of Cytochrome c Structural Fluctuation by Time-Resolved Proton Pulse

A brief proton pulse technique, based on photo-excitation of pyranine, was employed for measuring the protonation dynamics of cytochrome c in aqueous solutions. Time-resolved monitoring of the protonation state of the pyranine anion yielded detailed information from which the temporal state protonation of the surface carboxylates and histidine residues were deduced. The surface groups were found to be coupled by electrostatic interaction where the state of protonation of one affects the pK of the others. In the same sense, altering the charge distribution of a protein by a redox reaction affects the reactivity of the surface groups with protons in the bulk. The surface groups can exchange protons among themselves at a very high rate. The velocities of these reactions are functions of the connectivity between the proton binding sites. A single proton exchange reaction between the surface groups could be identified as the proton exchange between H26 and E44. The reaction is fast and affected by the redox state of the protein. It is proposed that the enhanced rate of this reaction is coupled with transient conformational changes that are not noticeable in time-averaging measurements such as X-ray diffraction or NMR.     

Effect of decortication and protease treatment on the kinetics of liquefaction,saccharification, and ethanol production from sorghum

BACKGROUND: This study analyzes the effect of decortication and protease treatment on the kinetics of liquefaction, saccharification and ethanol production from sorghum kernels. In general, bioethanol yields from sorghum are lower than those from maize. This has been attributed to reduced access of starch‐degrading enzymes due to the crosslinked protein net in the sorghum kernels. RESULTS: Liquefaction is described as a zero order kinetics process, with reaction rates enhanced by protease treatment. The use of protease almost doubled the liquefaction rate in both whole and decorticated sorghum, compared with untreated kernels. During saccharification of decorticated sorghum, protease treatment significantly affected the glucose/starch yield and the glucose concentration profile over time. When compared with maize, protease treatment of decorticated sorghum resulted in superior ethanol production rates. Specific ethanol yields during fermentation were statistically comparable with those for maize. CONCLUSION: Protease treatment of decorticated sorghum kernels can impart substantial economic benefits in terms of improvement of bioethanol yield (13% over whole sorghum) and in reduced fermentation time (approximately 50% with respect to maize). Copyright © 2010 Society of Chemical Industry     

Calcium phosphate substrates with emulsion-derived roughness: Processing,characterisation and interaction with human mesenchymal stem cells

Calcium phosphates (CaP) have been the subject of several studies that often lack a systematic approach to understanding how their properties affect biological response. CaP particles functionalised with a pH-responsive polymer (BCS) were used to prepare microporous substrates (porosity between 70 and 75% and pore sizes of 5–20 μm) through the aggregation of oil-in-water emulsions by controlling solid loading, emulsification energy, pH, drying and sintering conditions. The combined effect of surface roughness (roughness amplitude, R_a between 0.9–1.7 μm) and chemistry (varying Hydroxyapatite/β-Tricalcium phosphate ratio) on human mesenchymal stem cells was evaluated. HA substrates stimulated higher cell adhesion and proliferation (especially with lower R_a), but cell area increased with β-TCP content. The effect of surface roughness depended of chemistry: HA promoted higher mineralising activity when R_a ～ 1.5 μm, whereas β-TCP substrates stimulated a more osteogenic profile when R_a ～ 1.7 μm. A novel templating method to fabricate microporous CaP substrates was developed, opening possibilities for bone substitutes with controlled features.     

Cellular Integrin α5β1 and Exosomal ADAM17 Mediate the Binding and Uptake of Exosomes Produced by Colorectal Carcinoma Cells

Approximately 25% of colorectal cancer (CRC) patients develop peritoneal metastasis, a condition associated with a bleak prognosis. The CRC peritoneal dissemination cascade involves the shedding of cancer cells from the primary tumor, their transport through the peritoneal cavity, their adhesion to the peritoneal mesothelial cells (PMCs) that line all peritoneal organs, and invasion of cancer cells through this mesothelial cell barrier and underlying stroma to establish new metastatic foci. Exosomes produced by cancer cells have been shown to influence many processes related to cancer progression and metastasis. In epithelial ovarian cancer these extracellular vesicles (EVs) have been shown to favor different steps of the peritoneal dissemination cascade by changing the functional phenotype of cancer cells and PMCs. Little is currently known, however, about the roles played by exosomes in the pathogenesis and peritoneal metastasis cascade of CRC and especially about the molecules that mediate their interaction and uptake by target PMCs and tumor cells. We isolated exosomes by size−exclusion chromatography from CRC cells and performed cell-adhesion assays to immobilized exosomes in the presence of blocking antibodies against surface proteins and measured the uptake of fluorescently-labelled exosomes. We report here that the interaction between integrin α5β1 on CRC cells (and PMCs) and its ligand ADAM17 on exosomes mediated the binding and uptake of CRC-derived exosomes. Furthermore, this process was negatively regulated by the expression of tetraspanin CD9 on exosomes.     

Comparison of color gamuts among several types of paper with the same printing technology

In this work, we have studied the relationship among the colorimetric properties of different types of paper, having different finishing and grammage. Their color reproduction capability has also been analyzed by using the same printing technology (inkjet printing). On the one hand, we have plotted CIELAB data under the illuminant D50 into constant lightness and hue‐angle planes to be compared with MacAdam limits and with Pointer's real‐world surface color. On the other hand, we have calculated the volume gamut of the color solid associated to each color paper gamut. Analyzing the results, we have checked that there is not any clear relationship among the colorimetric properties of paper (for instance, CIE whiteness index, etc.) and the color gamut volume associated. However, the colorimetric parameters associated to the printed sample showed a quite good linear correlation between the minimum lightness (or the maximum blackness value) of the printed color chart and the color gamut volume. In particular, the greatest color gamut volume corresponds to the glossy papers taking into account this correlation for inkjet printing. © 2009 Wiley Periodicals, Inc. Col Res Appl, 34, 330–336, 2009     

Intraindividual variability is related to cognitive change in older adults: Evidence for within-person coupling.

In this study, the authors addressed the longitudinal nature of intraindividual variability over 3 years. A sample of 304 community-dwelling older adults, initially between the ages of 64 and 92 years, completed 4 waves of annual testing on a battery of accuracy- and latency-based tests covering a wide range of cognitive complexity. Increases in response-time inconsistency on moderately and highly complex tasks were associated with increasing age, but there were significant individual differences in change across the entire sample. The time-varying covariation between cognition and inconsistency was significant across the 1-year intervals and remained stable across both time and age. On occasions when intraindividual variability was high, participants' cognitive performance was correspondingly low. The strength of the coupling relationship was greater for more fluid cognitive domains such as memory, reasoning, and processing speed than for more crystallized domains such as verbal ability. Variability based on moderately and highly complex tasks provided the strongest prediction. These results suggest that intraindividual variability is highly sensitive to even subtle changes in cognitive ability. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Representation of Women in News and Photos: Comparing Content to Perceptions

This study uses a feminist framework of masculine cultural hegemony to examine the representation of women in two newspapers—a medium-sized newspaper (Study 1) and a larger newspaper (Study 2). Surveys gauged news staff and news reader perceptions of female representation in news content to determine if perceptions matched content-analysis findings. Content-analysis results revealed a greater proportion of males than females in both content and photos for both studies. Female staffers were more likely than their male counterparts to perceive this disparity, and news readers perceived an even greater disparity than the news staff. News staffs were less likely to perceive disparities in the editorial, travel, and entertainment sections. We discuss the findings in relation to cultural hegemony and the representation of women in the news.     

Photocatalytic oxidation of aliphatic and aromatic sulfides in the presence of silica adsorbed or zeolite-encapsulated 2,4,6-triphenyl(thia)pyrylium

Solid photocatalysts in which the 2,4,6-triphenylpyrylium (TP) or 2,4,6-triphenylthiapyrylium cation (TTP) are encapsulated within zeolite Y or deposited on mesoporous silica are used as photosensitizers for the oxidation of sulfides, both aromatic and aliphatic, in solvents of various polarity. Contrary to the same cations in solution, these solid sensitizers are not significantly degraded during operation. An effective oxygenation takes place leading to sulfoxides, disulfides, sulfinic and sulfenic esters as well as sulfonic acids. This large scope method shows a limited dependence on the substrate structure and on conditions and is suitable for the abatement of sulfur-containing pollutants.