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.     

Trifluoromethylcinnamanilide Michael Acceptors for Treatment of Resistant Bacterial Infections

A series of thirty-two anilides of 3-(trifluoromethyl)cinnamic acid (series 1) and 4-(trifluoromethyl)cinnamic acid (series 2) was prepared by microwave-assisted synthesis. All the compounds were tested against reference strains Staphylococcus aureus ATCC 29213 and Enterococcus faecalis ATCC 29212 and resistant clinical isolates of methicillin-resistant S. aureus (MRSA) and vancomycin-resistant E. faecalis (VRE). All the compounds were evaluated in vitro against Mycobacterium smegmatis ATCC 700084 and M. marinum CAMP 5644. (2E)-3-[3-(Trifluoromethyl)phenyl]-N-[4-(trifluoromethyl)phenyl]prop-2-enamide (1j), (2E)-N-(3,5-dichlorophenyl)-3-[3-(trifluoromethyl)phenyl]prop-2-enamide (1o) and (2E)-N-[3-(trifluoromethyl)phenyl]-3-[4-(trifluoromethyl)-phenyl]prop-2-enamide (2i), (2E)-N-[3,5-bis(trifluoromethyl)phenyl]-3-[4-(trifluoromethyl)phenyl]-prop-2-enamide (2p) showed antistaphylococcal (MICs/MBCs 0.15–5.57 µM) as well as anti-enterococcal (MICs/MBCs 2.34–44.5 µM) activity. The growth of M. marinum was strongly inhibited by compounds 1j and 2p in a MIC range from 0.29 to 2.34 µM, while all the agents of series 1 showed activity against M. smegnatis (MICs ranged from 9.36 to 51.7 µM). The performed docking study demonstrated the ability of the compounds to bind to the active site of the mycobacterial enzyme InhA. The compounds had a significant effect on the inhibition of bacterial respiration, as demonstrated by the MTT assay. The compounds showed not only bacteriostatic activity but also bactericidal activity. Preliminary in vitro cytotoxicity screening was assessed using the human monocytic leukemia cell line THP-1 and, except for compound 2p, all effective agents did show insignificant cytotoxic effect. Compound 2p is an interesting anti-invasive agent with dual (cytotoxic and antibacterial) activity, while compounds 1j and 1o are the most interesting purely antibacterial compounds within the prepared molecules.     

Merging high doxorubicin loading with pronounced magnetic response and bio-repellent properties in hybrid drug nanocarriers

Hybrid magnetic drug nanocarriers are prepared via a self-assembly process of poly(methacrylic acid)-graft-poly(ethyleneglycol methacrylate) (p(MAA-g-EGMA)) on growing iron oxide nanocrystallites. The nanocarriers successfully merge together bio-repellent properties, pronounced magnetic response, and high loading capacity for the potent anticancer drug doxorubicin (adriamicin), in a manner not observed before in such hybrid colloids. High magnetic responses are accomplished by engineering the size of the magnetic nanocrystallites (～13.5 nm) following an aqueous single-ferrous precursor route, and through adjustment of the number of cores in each colloidal assembly. Complementing conventional magnetometry, the magnetic response of the nanocarriers is evaluated by magnetophoretic experiments providing insight into their internal organization and on their response to magnetic manipulation. The structural organization of the graft-copolymer, locked on the surface of the nanocrystallites, is further probed by small-angle neutron scattering on single-core colloids. Analysis showed that the MAA segments selectively populate the area around the magnetic nanocrystallites, while the poly(ethylene glycol)-grafted chains are arranged as protrusions, pointing towards the aqueous environment. These nanocarriers are screened at various pHs and in highly salted media by light scattering and electrokinetic measurements. According to the results, their stability is dramatically enhanced, as compared to uncoated nanocrystallites, owing to the presence of the external protective PEG canopy. The nanocarriers are also endowed with bio-repellent properties, as evidenced by stability assays using human blood plasma as the medium.     

3D woven green composites from textile waste: mechanical performance

Application of textile waste for development of value added green composites has been carried out in this work. Textile fabric waste is collected from various sources. These waste materials are garneted, so as to produce loose fibrous material, subsequently this fibrous material was converted into twisted strand for manufacturing of 3D woven preforms for production of composites. Twisted strands are converted into orthogonal 3D woven structure. The fibers extracted from waste material are combined with polypropylene in 60/40 proportion. Composites of various specifications are developed to examine their end-use applications. These composite materials are characterized for their mechanical behavior to find out the response against tensile loading, flexural stress, and impact force. The effects of moisture absorption on mechanical properties of composites are investigated. 3D woven fabric reinforced composites produced by using waste fiber yarn and normal cotton OE yarn do not exhibit any significant difference in the mechanical behavior of composite. This result confirmed that waste material can be safely used as reinforcing structure in green composite manufacturing.     

Cationization of cellulose fibers for composites

In this work, composites were made by improved bonding of the cationized cellulosic fabric and some anionic matrix like Carboxymethyl Cellulose (CMC). Mercerized cotton fabric was cationized using cationization agent, TEXAMIN ECE with varying percentage from 1 to 10%. Those cationized cotton samples were incorporated in anionic matrices and the resultant composites were tested for mechanical properties on Universal Textile Tensile Testing machine. In the case of CMC as the matrix for the cationized reinforcement, the increasing trend in the tensile strength and elongation at break was observed. Scanning Electron Microscope images showed no extraordinary changes in the physical appearance of the cationized samples. Thermograms derived from Differential Scanning Calorimetry were informative for the evaluation of the thermal behavior of composites.     

ExPlanTech: Applying multi-agent systems in production planning

This paper presents work carried out within the 'ExPlanTech' project (IST-1999-20171) funded in part by the European Commission's Information Technologies Programme. The mission of the ExPlanTech technology transfer project is to introduce, customize and exploit the multi-agent production planning technology (ProPlanT multi-agent system research prototype) in two specific industrial enterprises. An agent-driven service negotiations and decision process, based on usagecentred knowledge about task requirements, substitutes the traditional production planning activity. We introduce a methodology for integration of the projectdriven production planning based on agent-based engineering within the existing enterprise resource planning system. This novel production planning technology will facilitate optimization of resource utilization and supplier chain while meeting the customer demands. This paper describes a FIPA-compliant implementation of the ExPlanTech technology at the LIAZ Pattern Shop manufacturing company. We describe the structure of the agent community, types of agents, implementation of the planning strategy and its incorporation within the real production environment.     

Study of TATP: Influence of Reaction Conditions on Product Composition

The influence of reaction conditions (temperature, catalyst type, catalyst concentration – represented as molar ratio catalyst/acetone n_c/n_a) on the composition of the product formed from the reaction of acetone and hydrogen peroxide (30%) under acid catalysis was studied. 3,3,6,6,9,9‐Hexamethyl‐1,2,4,5,7,8‐hexoxonane (TATP) was found to be the major product when the content of the catalyst in the reaction mixture is low (n_c/n_a ≤ 0.5). A single side product (peak 10.2) in an amount ranging from 1.5 to 8% of the total peak area was present in all the prepared samples. Three other side products were found when catalyzing by hydrochloric and nitric acids. Temperature and catalyst type did not have a significant influence on the composition of the product at low catalyst concentration. Increasing the catalyst concentration led to the formation of 3,3,6,6‐tetramethyl‐1,2,4,5‐tetroxane (DADP) either as a co‐product of TATP or as an exclusive product depending on the concentration of the catalyst.     

Two-Dimensional Modeling of Self-Propagating High-Temperature Synthesis of Strontium-Doped Lanthanum Manganate

A two-dimensional finite element model is developed to study the reaction kinetics and heat transfer during the self-propagating high-temperature synthesis of La_0.6Sr_0.4MnO₃, a cathode and interconnect material used in solid oxide fuel cells. The activation energy of La_0.6Sr_0.4MnO₃ formation was calculated from experimental temperature history. The calculated spatial-temporal temperature profile, heat generation rate, reaction conversion, and flow pattern of surrounding gas during the reaction are reported in this work. Hot spots are found at the corner near the ignition point shortly after the ignition. The model provided a simple and reliable way to design a large-scale production of La_0.6Sr_0.4MnO₃.     

Colorimetric method for phase evaluation

Measurements of very small phase changes in optical measurement techniques are usually performed with interferometric methods that are based on evaluation of interference patterns, which correspond to a phase change of the investigated wave field. If values of the phase change are small, it is difficult to determine accurately the phase values, and one needs very expensive measurement systems. We present a simple method for evaluation of small phase variations that uses the interference of polychromatic light. The phase change affects the color of the interference pattern, and the color of the interference pattern corresponds to a specific phase change that can be evaluated using colorimetric analysis. We describe and analyze our colorimetric phase evaluation method. The proposed method offers accurate results, and it may be suitable for practical utilization in the optical industry.     

Dependence of Strehl ratio on f-number of optical system

Formulas for a minimum of wave aberration variance and a maximum of the Strehl ratio in the optimal image point are derived using the third- and fifth-order aberration theory. Moreover, relations for the calculation of the optimal value of f-number of the optical system were derived, which enabled us to theoretically analyze real optical systems and their image quality. The optimal f-number corresponds to such a value of f-number when the image quality of a real optical system is comparable to an aberration-free optical system. This value may also serve as an auxiliary criterion of the image quality of the optical system, for example, in photography.