Modular approach to select bacteriophages targeting Pseudomonas aeruginosa for their application to children suffering with cystic fibrosis

This review discusses the potential application of bacterial viruses (phage therapy) towards the eradication of antibiotic resistant Pseudomonas aeruginosa in children with cystic fibrosis (CF). In this regard, several potential relationships between bacteria and their bacteriophages are considered. The most important aspect that must be addressed with respect to phage therapy of bacterial infections in the lungs of CF patients is in ensuring the continuity of treatment in light of the continual occurrence of resistant bacteria. This depends on the ability to rapidly select phages exhibiting an enhanced spectrum of lytic activity among several well-studied phage groups of proven safety. We propose a modular based approach, utilizing both mono-species and hetero-species phage mixtures. With an approach involving the visual recognition of characteristics exhibited by phages of well-studied phage groups on lawns of the standard P. aeruginosa PAO1 strain, the simple and rapid enhancement of the lytic spectrum of cocktails is permitted, allowing the development of tailored preparations for patients capable of circumventing problems associated with phage resistant bacterial mutants

The T7-Related Pseudomonas putida Phage ϕ15 Displays Virion-Associated Biofilm Degradation Properties

Formation of a protected biofilm environment is recognized as one of the major causes of the increasing antibiotic resistance development and emphasizes the need to develop alternative antibacterial strategies, like phage therapy. This study investigates the in vitro degradation of single-species Pseudomonas putida biofilms, PpG1 and RD5PR2, by the novel phage ϕ15, a ‘T7-like virus’ with a virion-associated exopolysaccharide (EPS) depolymerase. Phage ϕ15 forms plaques surrounded by growing opaque halo zones, indicative for EPS degradation, on seven out of 53 P. putida strains. The absence of haloes on infection resistant strains suggests that the EPS probably act as a primary bacterial receptor for phage infection. Independent of bacterial strain or biofilm age, a time and dose dependent response of ϕ15-mediated biofilm degradation was observed with generally a maximum biofilm degradation 8 h after addition of the higher phage doses (104 and 106 pfu) and resistance development after 24 h. Biofilm age, an in vivo very variable parameter, reduced markedly phage-mediated degradation of PpG1 biofilms, while degradation of RD5PR2 biofilms and ϕ15 amplification were unaffected. Killing of the planktonic culture occurred in parallel with but was always more pronounced than biofilm degradation, accentuating the need for evaluating phages for therapeutic purposes in biofilm conditions. EPS degrading activity of recombinantly expressed viral tail spike was confirmed by capsule staining. These data suggests that the addition of high initial titers of specifically selected phages with a proper EPS depolymerase are crucial criteria in the development of phage therapy

High-Temperature Oxidation of High-Entropic Alloys: A Review

Over the past few years, interest in high-entropic alloys (HEAs) has been growing. A large body of research has been undertaken to study aspects such as the microstructure features of HEAs of various compositions, the effect of the content of certain elements on the mechanical properties of HEAs, and, of course, special properties such as heat resistance, corrosion resistance, resistance to irradiation with high-energy particles, magnetic properties, etc. However, few works have presented results accumulated over several years, which can complicate the choice of directions for further research. This review article presents the results of studies of the mechanisms of high-temperature oxidation of HEAs of systems: Al-Co-Cr-Fe-Ni, Mn-Co-Cr-Fe-Ni, refractory HEAs. An analysis made it possible to systematize the features of high-temperature oxidation of HEAs and propose new directions for the development of heat-resistant HEAs. The presented information may be useful for assessing the possibility of the practical application of HEAs in the aerospace industry, in nuclear and chemical engineering, and in new areas of energy

Al_0.25CoCrFeNiV High Entropy Alloy Coating Deposited by Laser Cladding on Stainless Steel

This paper studies the microstructure, composition and properties of a Al0.25CoCrFeNiV high entropy alloy coating (HEAC) deposited by laser cladding on austenitic-grade stainless steel. Laser cladding was carried out in an argon atmosphere on a FL-Clad-R-4 laser metal deposition complex with the following parameters: the laser power was 1400 W, the spot diameter was 3 mm, the track displacement was 1.2 mm, and the scanning speed was set to 10 mm/s. A change in the microstructure of the coating after laser cladding was revealed in comparison with as-cast high entropy alloy (HEA) Al0.25CoCrFeNiV. A significant decrease was found in the size of vanadium precipitates, from 20–40 µm in the as-cast state to 1–3 µm after laser cladding. A change in microhardness over the thickness of the coating from 370 HV0.3 at the outer surface to 270 HV0.3 at the boundary with the substrate was established, which may be due to the diffusion of Fe from the stainless steel into the coating material during laser cladding. Despite these features, the resulting coating adheres tightly to the substrate, and has no cracks or other defects, which indicates the possibility of using laser cladding to create coatings from high entropy alloys

Study of the Influence of V, Mo and Co Additives on the Carbide Formation and Microhardness during Thermal Diffusion Chrome Planting of X35CrNi2-3 Steel

Saturation diffusion with chromium has not been adequately studied among all the surface thermochemical treatment (STCT) processes of steels. Especially, the complex saturation behavior when several elements are added directly for chemical treatment needs to be systematically studied. This work aims at determining the effect of V, Mo, and Co on the parameters of chromium thermal saturation diffusion (thickness, phase composition, microstructure, and microhardness) of the surface layer in X35CrNi2-3 steel. The process was carried out at a temperature of 1000 °C for 24 h. The results showed that complex structural chromium plating together with the addition of strong carbide-forming elements (V and Mo) has a significant effect on the phase composition of the fabricated layer, where the formation of VC and Mo2C carbides significantly increases the microhardness of the samples to 2000 HV and 2500 HV, respectively. On the other hand, the addition of Co with a less carbide-forming affinity has little effect on the phase composition of the coating, and nitride compounds predominated in the microstructure similar to the single-element chromium plating. The results indicate the possibility of improving and accelerating the traditional thermal chromium plating processes and opening up new horizons for obtaining gradient coatings with superior tribological properties

Effect of Electromagnetic Pulses on the Microstructure and Abrasive Gas Wear Resistance of Al0.25CoCrFeNiV High Entropy Alloy

High entropy alloys (HEAs) are among the most promising materials, owing to their vast chemical composition window and unique properties. Segregation is a well-known phenomenon during the solidification of HEAs, which negatively affects their properties. The electromagnetic pulse (EMP) is a new technique for the processing of a metal melt that can hinder segregation during solidification. In this study, the effect of an EMP on the microstructure and surface properties of Al0.25CoCrFeNiV HEA is studied. An EMP, with an amplitude of 10 kV, a leading edge of 0.1 ns, a pulse duration of 1 ns, a frequency of 1 kHz, and pulse power of 4.5 MW, was employed for melt treatment. It was found that the microstructure of Al0.25CoCrFeNiV HEA changes significantly from dendritic, for an untreated sample, to lamellar “pearlite-like”, for an EMP treated sample. Moreover, EMPs triggered the formation of a needle-like σ-phase within the solid solution grains. Finally, these microstructural and compositional changes significantly increased the microhardness of Al0.25CoCrFeNiV HEA, from 343 ± 10 HV0.3 (without the EMP) to 553 ± 15 HV0.3 (after the EMP), and improved its resistance against gas-abrasive wear. Finally, an EMP is introduced as an effective route to modify the microstructure and phase formation of cast HEAs, which, in turn, opens up broad horizons for fabricating cast samples with tailorable microstructures and improved properties

Phage phiKZ—The First of Giants

The paper covers the history of the discovery and description of phiKZ, the first known giant bacteriophage active on Pseudomonas aeruginosa. It also describes its unique features, especially the characteristic manner of DNA packing in the head around a cylinder-shaped structure (“inner body”), which probably governs an ordered and tight packaging of the phage genome. Important properties of phiKZ-like phages include a wide range of lytic activity and the blue opalescence of their negative colonies, and provide a background for the search and discovery of new P. aeruginosa giant phages. The importance of the phiKZ species and of other giant phage species in practical phage therapy is noted given their broad use in commercial phage preparations

Genome Instability of Pseudomonas aeruginosa phages of the EL species: examination of virulent mutants

The article continues a study of pseudolysogeny in Pseudominas aeruginosa infected with phiKZ-like phages of the EL species. Analysis was performed for several newly isolated vir mutants of EL phages (EL and RU) that were virulent (capable of causing lysis of bacteria infected with the wild-type phage) and a lower extent of opalescence of negative colonies (NCs). Wile-type recombinants were detected in crosses of virulent mutants of phages EL and RU to confirm the polygenic control of virulence. Since a deletion mutation was found in one of the virulent EL mutants and high genetic instability was characteristic of another mutant, a mobile genetic element was assumed to play a role in mutagenesis. Pseudolysogeny of bacteria provides for horizontal gene transfer between different bacterial strains. Hence, sequencing of the phage genome and demonstration of the lack of toxic gene products are insufficient for the phage to be included into a therapeutic mixture. To use live phages, it is essential to study in detail the possible consequences of their interaction with host bacteria. © 2011 Pleiades Publishing, Ltd.status: publishe

A Study of the Structural Characteristics of Titanium Alloy Products Manufactured Using Additive Technologies by Combining the Selective Laser Melting and Direct Metal Deposition Methods

Titanium alloy product manufacturing is traditionally considered to be a rather difficult task. Additive manufacturing technologies, which have recently become quite widespread, can ensure the manufacture of titanium alloys products of an arbitrary geometrical shape. During this study, we have developed a methodology for manufacturing titanium alloys products using additive technologies on FL-Clad-R-4 complex of laser melting of metals by combined Selective Laser Melting (SLM) and Direct Metal Deposition (DMD) methods. Ti–6Al–4V and Ti–6Al–4Mo–1V alloys were used for the manufacture of samples. We studied the microstructure of the obtained details and measured the microhardness of the samples. We discovered a gradient of the structure throughout the height of the details walls, which is connected with the peculiarities of thermal cycles of the technology used. This affected the microhardness values: in the upper part of the details, the microhardness is 10–25% higher (about 500 HV) than in the lower part (about 400 HV). Products made according to the developed technique do not have visible defects and pores. The obtained results indicate the competitiveness of the proposed methodology