Dual Escherichia coli DNA Gyrase A and B Inhibitors with Antibacterial Activity

The emergence of multidrug-resistant bacteria is a global health threat necessitating the discovery of new antibacterials and novel strategies for fighting bacterial infections. We report first-in-class DNA gyrase B (GyrB) inhibitor/ciprofloxacin hybrids that display antibacterial activity against Escherichia coli. Whereas DNA gyrase ATPase inhibition experiments, DNA gyrase supercoiling assays, and in vitro antibacterial assays suggest binding of the hybrids to the E. coli GyrA and GyrB subunits, an interaction with the GyrA fluoroquinolone-binding site seems to be solely responsible for their antibacterial activity. Our results provide a foundation for a new concept of facilitating entry of nonpermeating GyrB inhibitors into bacteria by conjugation with ciprofloxacin, a highly permeable GyrA inhibitor. A hybrid molecule containing GyrA and GyrB inhibitor parts entering the bacterial cell would then elicit a strong antibacterial effect by inhibition of both the GyrA and GyrB subunits of DNA gyrase and potentially slow bacterial resistance development.     

Determination of flow stress properties of machinable materials with help of simple compression and orthogonal machining test

This paper describes a possible method for predicting values of orthogonal metal cutting properties such as shear angle, cutting force etc., on a basis of the well known Hollomon equation, using a simple compression test in order to avoid any cutting experiments. There are two possibilities: the flow stress properties can be obtained from an independent material test; or by measuring the active and passive cutting forces from the orthogonal machining test itself. This paper is concerned with a material flow stress equation, including the effects of strain (ε), strain rate ( ) and temperature (T), which is one of the five equations that have to be solved in simulation analysis with the finite element method. In finding a solution for those five equations, it is necessary to dispose of flow stress properties by rearrangement of the Hollomon equation and so making it usable for cutting process investigation. The rearrangement is described in this paper.     

Microstructure Development in Low-Antimony Oxide-Doped Zinc Oxide Ceramics

The grain growth of ZnO ceramics sintered with low additions of Sb₂O₃ (<500 ppm of Sb) was investigated. Additions of Sb<250 ppm resulted in a coarse-grained microstructure with large ZnO grains (55–70 μm), much larger than the grain size of ZnO ceramics without any Sb₂O₃ addition (45 μm). The addition of 500 ppm of Sb resulted in a fine-grained microstructure with an average ZnO grain size of about 12 μm. The results are explained by an inversion-boundary (IB) -induced grain-growth mechanism. The grain-growth exponent has a value of about 2 as long as the grains containing IBs grow at the expense of IB-free grains. It increases to about 4 after the IB-containing grains impinge on each other, and achieves values above 10 for additions of 500 ppm of Sb when IBs nucleate in nearly all the ZnO grains so that grains with IBs prevail in the microstructure at an early stage in the grain-growth process.     

Intracellular Trafficking of Cationic Carbon Dots in Cancer Cell Lines MCF-7 and HeLa—Time Lapse Microscopy,Concentration-Dependent Uptake,Viability, DNA Damage,and Cell Cycle Profile

Fluorescent carbon dots (CDs) are potential tools for the labeling of cells with many advantages such as photostability, multicolor emission, small size, rapid uptake, biocompatibility, and easy preparation. Affinity towards organelles can be influenced by the surface properties of CDs which affect the interaction with the cell and cytoplasmic distribution. Organelle targeting by carbon dots is promising for anticancer treatment; thus, intracellular trafficking and cytotoxicity of cationic CDs was investigated. Based on our previous study, we used quaternized carbon dots (QCDs) for treatment and monitoring the behavior of two human cancer cell MCF-7 and HeLa lines. We found similarities between human cancer cells and mouse fibroblasts in the case of QCDs uptake. Time lapse microscopy of QCDs-labeled MCF-7 cells showed that cells are dying during the first two hours, faster at lower doses than at higher ones. QCDs at a concentration of 100 µg/mL entered into the nucleus before cellular death; however, at a dose of 200 µg/mL, blebbing of the cellular membrane occurred, with a subsequent penetration of QCDs into the nuclear area. In the case of HeLa cells, the dose-depended effect did not happen; however, the labeled cells were also dying in mitosis and genotoxicity occurred nearly at all doses. Moreover, contrasted intracellular compartments, probably mitochondria, were obvious after 24 h incubation with 100 µg/mL of QCDs. The levels of reactive oxygen species (ROS) slightly increased after 24 h, depending on the concentration, thus the genotoxicity was likely evoked by the nanomaterial. A decrease in viability did not reach IC 50 as the DNA damage was probably partly repaired in the prolonged G0/G1 phase of the cell cycle. Thus, the defects in the G2/M phase may have allowed a damaged cell to enter mitosis and undergo apoptosis. The anticancer effect in both cell lines was manifested mainly through genotoxicity.     

Why Monoamine Oxidase B Preferably Metabolizes N-Methylhistamine over Histamine: Evidence from the Multiscale Simulation of the Rate-Limiting Step

Histamine levels in the human brain are controlled by rather peculiar metabolic pathways. In the first step, histamine is enzymatically methylated at its imidazole N^τ atom, and the produced N-methylhistamine undergoes an oxidative deamination catalyzed by monoamine oxidase B (MAO-B), as is common with other monoaminergic neurotransmitters and neuromodulators of the central nervous system. The fact that histamine requires such a conversion prior to oxidative deamination is intriguing since MAO-B is known to be relatively promiscuous towards monoaminergic substrates; its in-vitro oxidation of N-methylhistamine is about 10 times faster than that for histamine, yet this rather subtle difference appears to be governing the decomposition pathway. This work clarifies the MAO-B selectivity toward histamine and N-methylhistamine by multiscale simulations of the rate-limiting hydride abstraction step for both compounds in the gas phase, in aqueous solution, and in the enzyme, using the established empirical valence bond methodology, assisted by gas-phase density functional theory (DFT) calculations. The computed barriers are in very good agreement with experimental kinetic data, especially for relative trends among systems, thereby reproducing the observed MAO-B selectivity. Simulations clearly demonstrate that solvation effects govern the reactivity, both in aqueous solution as well as in the enzyme although with an opposing effect on the free energy barrier. In the aqueous solution, the transition-state structure involving histamine is better solvated than its methylated analog, leading to a lower barrier for histamine oxidation. In the enzyme, the higher hydrophobicity of N-methylhistamine results in a decreased number of water molecules at the active side, leading to decreased dielectric shielding of the preorganized catalytic electrostatic environment provided by the enzyme. This renders the catalytic environment more efficient for N-methylhistamine, giving rise to a lower barrier relative to histamine. In addition, the transition state involving N-methylhistamine appears to be stabilized by the surrounding nonpolar residues to a larger extent than with unsubstituted histamine, contributing to a lower barrier with the former.     

Modified firefly algorithm using quaternion representation

Quaternions are a number system, which extends complex numbers. They are especially useful in areas where fast rotation calculations are needed, e.g., programming video games or controllers of spacecraft. This paper proposes to use quaternion for the representation of individuals in firefly algorithm so as to enhance the performance of the firefly algorithm and to avoid any stagnation. The preliminary results of our experiments after optimizing a test-suite consisting of ten standard functions, showed that the proposed firefly algorithms using quaternion’s representation improved the results of the original firefly algorithm.     

A numerical model for the analyses of heat transfer and leakages in a rotary air preheater

Air preheaters make a considerable contribution to the improved overall efficiency of fossil-fuel-fired power plants. In this study we used a combination of fluid dynamics and a newly developed three-dimensional numerical model for heat transfer as the basis for a theoretical analysis of a rotary air preheater. The model enables studies of the flue-gas flow through the preheater and the adjoining channels as well as the regenerative heat transfer and the resulting temperature distribution in the matrix of the preheater. Special attention was focused on the influences of leakages on the flue-gas parameters in the preheater. The numerical analysis and the experimental results showed an obvious dependence of the flue-gas parameters on various seal settings. Based on the results a method for online monitoring of the tightness of the radial seals is proposed.     

Zero—a blend of static typing and dynamic metaprogramming

Zero is an experimental statically typed, fully object-oriented reflective programming language. Reflective features cover introspection as well as structural and behavioural reflection. The reflective facilities include safe method and class replacements and detailed modification of methods. These enable Zero programs to quickly accommodate to run-time requirements. Behavioural reflection is realised using handlers (hooks), which may be attached to all language constructs based on closures. Zero provides an efficient static typing system with run-time extensions. Methods are first class values and are represented as objects when such representation is required. By using such representation, Zero provides elegant use of statically typed higher-order methods.     

Deformation of specimen during laser surface remelting

During laser surface remelting, thermal expansion in heating, and contraction of material in cooling, and due to microstructure changes in the heated surface layer, the specimen deforms. This induces volume changes during the process of remelting the thin surface layer and causes internal stresses. By measuring the deformation of the specimen during the remelting process as well as after the laser beam interaction, i.e., to the moment when the specimen cools down to the ambient temperature, it is possible to follow the progress of deformation in the specimen material. Detailed information on the deformation events in the specimen material during the laser remelting process of a thin surface layer enables the engineer to optimize the process variables in terms of amount of deformation or magnitude of residual stresses on completion of cooling.