Structural Analysis of the Ancestral Haloalkane Dehalogenase AncLinB-DmbA

Haloalkane dehalogenases (EC 3.8.1.5) play an important role in hydrolytic degradation of halogenated compounds, resulting in a halide ion, a proton, and an alcohol. They are used in biocatalysis, bioremediation, and biosensing of environmental pollutants and also for molecular tagging in cell biology. The method of ancestral sequence reconstruction leads to prediction of sequences of ancestral enzymes allowing their experimental characterization. Based on the sequences of modern haloalkane dehalogenases from the subfamily II, the most common ancestor of thoroughly characterized enzymes LinB from Sphingobium japonicum UT26 and DmbA from Mycobacterium bovis 5033/66 was in silico predicted, recombinantly produced and structurally characterized. The ancestral enzyme AncLinB-DmbA was crystallized using the sitting-drop vapor-diffusion method, yielding rod-like crystals that diffracted X-rays to 1.5 Å resolution. Structural comparison of AncLinB-DmbA with their closely related descendants LinB and DmbA revealed some differences in overall structure and tunnel architecture. Newly prepared AncLinB-DmbA has the highest active site cavity volume and the biggest entrance radius on the main tunnel in comparison to descendant enzymes. Ancestral sequence reconstruction is a powerful technique to study molecular evolution and design robust proteins for enzyme technologies.     

Quercetin Ameliorates Testicular Damage in Zucker Diabetic Fatty Rats through Its Antioxidant,Anti-Inflammatory and Anti-Apoptotic Properties

The aim of this study was to investigate the effects of quercetin (QUE) on the testicular architecture as well as markers of oxidative, inflammatory, and apoptotic profile of male gonads in Zucker diabetic fatty (ZDF) rats suffering from Type 2 diabetes mellitus in the absence or presence of obesity. QUE was administered orally at a dose of 20 mg/kg/day for 6 weeks. Morphometric analysis revealed that QUE treatment led to an improvement in testicular appearance, particularly in the case of Obese ZDF rats. Furthermore, a significant stabilization of the antioxidant capacity (p < 0.05), superoxide dismutase and catalase activity (p < 0.01), with a concomitant decrease in lipid peroxidation (p < 0.05) were observed in Obese ZDF animals exposed to QUE. Our data also indicate a significant decline in the levels of interleukin (IL)-1 (p < 0.05), IL-6 (p < 0.01) and tumor necrosis factor alpha (p < 0.001) following QUE supplementation to Obese ZDF rats in comparison with their respective control. Finally, a significant down-regulation of the pro-apoptotic BAX protein (p < 0.0001) was observed in Obese ZDF rats administered with QUE, while a significant Bcl-2 protein overexpression (p < 0.0001) was recorded in Lean ZDF animals when compared to their untreated control. As such, our results suggest that QUE is a potentially beneficial agent to reduce testicular damage in ZDF rats with Type 2 diabetes mellitus by decreasing oxidative stress, chronic inflammation, and excessive cell loss through apoptosis.     

Effect of rapeseed oil on the rheological,mechanical and thermal properties of plastic lubricants

A strong difference in the physico-chemical properties of the plastic lubricants studied was found in this study through pressure drop, thermal analysis, vibration damping, texture hardness and rheological measurements. Oxidation aging of the lubricant sample containing rapeseed oil additive was proposed. Its higher thermal sensitivity was simultaneously confirmed by frequency dependent complex shear modulus of elasticity measurements as well as by rheological testing. Rapeseed oil modified lubricant showed a higher decrease in both storage and moduli losses due to a temperature increase from 16 to 26 ° C compared to the rapeseed oil free sample. Simultaneously, the flow curves were shifted to the higher shear stresses (for plastic lubricant without rapeseed oil additive) typical for rheopectic fluids. For the rapeseed oil modified lubricant, the flow curves were shifted to the lower shear stresses, indicating its thixotropic fluid behaviour. The synthetic lubricant without rapeseed oil additive exhibited higher dissipative rheological behaviour as reflected by decreasing first resonance frequency peak position compared to the rapeseed oil modified lubricant as obtained from vibration damping measurements. It was found that the synthetic lubricant exhibited better vibration damping properties and mechanical energy dissipation into heat due to its higher viscous friction than the rapeseed oil modified lubricant under experimental conditions.

     

Neuron Adhesion on Diamond: Competition between Polymer Treatment and Surface Morphology

The ability to form an efficient interface between material and neural cells is a crucial aspect for construction of neuroelectrodes. Diamond offers material characteristics that could, to a large extent, improve the performance of neuroelectrodes. The greatest advantage of diamond is a large variety of material and surface properties such as electrical conductivity, surface morphology, and surface chemistry. Such a variety of material characteristics can lead to various cellular responses. Here, the authors compare survival, adhesion, and neurite formation of primary neurons on diamond thin films of various morphologies and treatments with several types of polymers commonly used to enhance cell adhesion. The authors find that the variation of surface roughness of nanocrystalline diamond film when coated with polymer does not have a major influence on neuron survival or adhesion. The adhesion of neurons can be influenced by the selected type of polymer coating. High molecular weight of polyethylenimine results in lower viability, adhesion, and neurite formation. The addition of laminin to treated films do not lead to significant improvements in neuron adhesion and neurite development. Their findings emphasize the importance of the correct polymer treatment over morphological properties of diamond thin films as a material for forming interfaces with primary neurons.

     

Optomechanical Processing of Silver Colloids: New Generation of Nanoparticle–Polymer Composites with Bactericidal Effect

The properties of materials at the nanoscale open up new methodologies for engineering prospective materials usable in high-end applications. The preparation of composite materials with a high content of an active component on their surface is one of the current challenges of materials engineering. This concept significantly increases the efficiency of heterogeneous processes moderated by the active component, typically in biological applications, catalysis, or drug delivery. Here we introduce a general approach, based on laser-induced optomechanical processing of silver colloids, for the preparation of polymer surfaces highly enriched with silver nanoparticles (AgNPs). As a result, the AgNPs are firmly immobilized in a thin surface layer without the use of any other chemical mediators. We have shown that our approach is applicable to a broad spectrum of polymer foils, regardless of whether they absorb laser light or not. However, if the laser radiation is absorbed, it is possible to transform smooth surface morphology of the polymer into a roughened one with a higher specific surface area. Analyses of the release of silver from the polymer surface together with antibacterial tests suggested that these materials could be suitable candidates in the fight against nosocomial infections and could inhibit the formation of biofilms with a long-term effect.     

High-performance liquid chromatography study of stereospecific microsomal enzymes catalysing the reduction of a potential cytostatic drug, oracin. Interspecies comparison

One of the main metabolites of oracin (I) ?6-[2-(2-hydroxyethyl)aminoethyl]-5,11-dioxo-5,6-dihydro-11H-indeno[1,2- c] isoquinoline?, a potential cytostatic drug, is 11-dihydrooracin (II) ?(+),(-)-6-[2-(2-hydroxyethyl)aminoethyl]-5-oxo-11-hydroxy-5,6-dihydro-1 1H- indeno[1,2-c]isoquinoline?, a metabolite formed by the reduction of oracin's pro-chiral centre on C 11. This metabolite has been found in all laboratory species in vitro and in vivo and it constitutes the main metabolite in man. The stereospecificity of reducing enzymes participating in the oracin biotransformation pathway was investigated using microsomal preparations from standard laboratory animals. Enzyme stereospecificity has been defined as preferential formation by the enzyme of the (+) or (-) stereoisomer of II. Significant interspecies differences were observed in the stereospecificity of the respective biotransformation enzymes. HPLC quantitative determinations of both enantiomers were performed using a Chiralcel OD-R column as chiral stationary phase with excellent resolution and stability.     

Electronic interactions between "pea" and "pod": the case of oligothiophenes encapsulated in carbon nanotubes

One of the most challenging strategies to achieve tunable nanophotonic devices is to build robust nanohybrids with variable emission in the visible spectral range, while keeping the merits of pristine single-walled carbon nanotubes (SWNTs). This goal is realized by filling SWNTs ("pods") with a series of oligothiophene molecules ("peas"). The physical properties of these peapods are depicted by using aberration-corrected high-resolution transmission electron microscopy, Raman spectroscopy, and other optical methods including steady-state and time-resolved measurements. Visible photoluminescence with quantum yields up to 30% is observed for all the hybrids. The underlying electronic structure is investigated by density functional theory calculations for a series of peapods with different molecular lengths and tube diameters, which demonstrate that van der Waals interactions are the bonding mechanism between the encapsulated molecule and the tube.     

Development of colloidal carbon-based immunochromatographic strip for rapid detection of carbaryl in fruit juices

The differences in lipid content will lead to differences in protein structure and gel properties and it is a nice model system, so our study, based on the production process for Zhongxiang tofu, used soy protein isolate (SPI) as raw material and adopted circular dichroism and Raman spectrum to investigate the influence of different lipid content on protein secondary structure in tofu. Texture profile analysis and differential scanning calorimetry were used to analyze the influence of lipid on gel texture properties and state of water. The interactive mechanism at microscopic level was investigated by means of scanning electron microscopy. The results show that the additional lipid reacted with the soy protein and the lipid became a component of a newly formed molecular membrane. With an increase in added lipid, SPI gel hardness was reduced, while springiness increased. The proportion of α-helix and β-sheet in SPI gel at 16% (w/v) lipid are 4.0 and 26.0%, respectively. The equilibrium water content tended to decrease when lipid content was increased.