Helicobacter pylori virulence factors: subversion of host immune system and development of various clinical outcomes.

Helicobacter pylori (H. pylori) is a worldwide spread bacterium, co-evolving with humans for at least 100 000 years. Despite the uncertainty about the mode of H. pylori transmission, the development of intra-gastric and extra-gastric diseases is attributed to this bacterium. The morphological transformation and production of heterogenic virulence factors enable H. pylori to overcome the harsh stomach environment. Using numerous potent disease-associated virulence factors makes H. pylori a prominent pathogenic bacterium. These bacterial determinants are adhesins (e.g., blood group antigen-binding adhesin (BabA)/sialic acid-binding adhesin (SabA)), enzymes (e.g., urease), toxins (e.g., vacuolating cytotoxin A (VacA)), and effector proteins (e.g., cytotoxin-associated gene A (CagA)) involved in colonisation, immune evasion, and disease induction. H. pylori not only cleverly evades the immune system but also robustly induces immune responses. This insidious bacterium employs various strategies to evade human innate and adaptive immune responses, leading to a life-long infection. Owing to the alteration of surface molecules, innate immune receptors couldn't recognise this bacterium; moreover, modulation of effector T cells subverts adaptive immune response. Most of the infected humans are asymptomatic and only a few of them present severe clinical outcomes. Therefore, the identification of virulence factors will pave the way for the prediction of infection severity and the development of an effective vaccine. H. pylori virulence factors are hereby comprehensively reviewed and the bacterium evasion from the immune response is properly discussed.

Multipass Friction Stir Processing of Laser-Powder Bed Fusion AlSi10Mg: Microstructure and Mechanical Properties.

The effect of multipass friction stir processing (FSP) on the microstructure and mechanical properties of an AlSi10Mg alloy produced by laser-powder bed fusion was investigated. FSP was performed at a rotational speed of 950 rpm and traverse speed of 85 mm/min. The results indicated that FSP destroyed the coarse grain structure in the as-built AlSi10Mg by generating fine and equiaxed grain structures with shear texture components of A1*(111)[1¯1¯2] and A2*(111)[112¯], in addition to causing fragmentation and refinement of the Si networks. FSP reduced the tensile strength slightly but significantly improved ductility. One-pass FSP exhibited superior mechanical properties compared with the two- and three-pass scenarios. The higher strength of the one-pass sample was attributed to the strengthening mechanisms induced by the Si particles, which were grown by repeated FSP. The higher ductility of the one-pass sample was explained using the kernel and grain average misorientations. Furthermore, the post-FSP microstructural evolution and fracture behavior of the samples were discussed.

Grain Structure Formation and Texture Modification through Multi-Pass Friction Stir Processing in AlSi10Mg Alloy Produced by Laser Powder Bed Fusion.

A new strategy is proposed to modify the grain structure and crystallographic texture of laser-powder bed fusion AlSi10Mg alloy using multi-pass friction stir processing (FSP). Accordingly, 1-3 passes of FSP with 100% overlap were performed. Scanning electron microscopy and electron backscattered diffraction were used for microstructural characterization. Continuous dynamic recrystallization and geometric dynamic recrystallization are the governing mechanisms of grain refinement during FSP. The stir zones have bimodal grain structures containing large and fine grains. The multi-pass FSP caused a considerable increase in the volume fraction of the large-grained area in the stir zone, which contained higher values of low-angle boundaries and sharp shear texture components of B(11¯2)[110] and B¯(1¯12¯)[1¯1¯0]. The formation of low-energy grain boundaries in the stir zone and alignment of the low-energy crystallographic planes with the surface of the sample made the strategy of using multi-pass FSP a promising candidate for corrosion resistance enhancement in future studies. Moreover, the detailed evolution of the grains, texture components, grain boundaries, and Si particles is discussed.

Designing and development of epitope-based vaccines against Helicobacter pylori.

Helicobacter pylori infection is the principal cause of serious diseases (e.g. gastric cancer and peptic ulcers). Antibiotic therapy is an inadequate strategy in H. pylori eradication because of which vaccination is an inevitable approach. Despite the presence of countless vaccine candidates, current vaccines in clinical trials have performed with poor efficacy which makes vaccination extremely challenging. Remarkable advancements in immunology and pathogenic biology have provided an appropriate opportunity to develop various epitope-based vaccines. The fusion of proper antigens involved in different aspects of H. pylori colonization and pathogenesis as well as peptide linkers and built-in adjuvants results in producing epitope-based vaccines with excellent therapeutic efficacy and negligible adverse effects. Difficulties of the in vitro culture of H. pylori, high genetic variation, and unfavourable immune responses against feeble epitopes in the complete antigen are major drawbacks of current vaccine strategies that epitope-based vaccines may overcome. Besides decreasing the biohazard risk, designing precise formulations, saving time and cost, and induction of maximum immunity with minimum adverse effects are the advantages of epitope-based vaccines. The present article is a comprehensive review of strategies for designing and developing epitope-based vaccines to provide insights into the innovative vaccination against H. pylori.

Practical Methods for Expression of Recombinant Protein in the Pichia pastoris System.

One of the most critical challenges of genetic engineering is the expression of recombinant proteins using biological expression systems. Nowadays, different expression systems from bacteria to mammalian tissue culture cells are available for the production of recombinant proteins for medical and industrial purposes. Among various choices, yeast expression systems such as Pichia pastoris are promising candidates. The P. pastoris expression system is a standard tool for the production of biopharmaceuticals and industrial enzymes. It is also considered a unique host for the expression of subunit vaccines which could significantly affect the growing market of medical biotechnology. Using P. pastoris as an expression system for heterologous proteins, this article provides detailed basic protocols for cloning of a recombinant cassette into a suitable expression vector, the transformation of foreign vector DNAs into the yeast by electroporation, and expression and purification of desired recombinant protein. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Cloning of a recombinant cassette into a suitable expression vector Basic Protocol 2: Transformation of P. pastoris and selection of transformants Basic Protocol 3: Optimization and large-scale expression of recombinant proteins Basic Protocol 4: Purification of recombinant proteins.

Mycobacterium tuberculosis extracellular vesicles: exploitation for vaccine technology and diagnostic methods.

Tuberculosis (TB) is a fatal epidemic disease usually caused by Mycobacterium tuberculosis (Mtb). Pervasive latent infection, multidrug- and extensively drug-resistant tuberculosis (MDR- and XDR-TB), and TB/HIV co-infection make TB a global health problem, which emphasises the design and development of efficient vaccines and diagnostic biomarkers. Extracellular vesicles (EVs) secretion is a conserved phenomenon in all the domains of life. Various cargos such as nucleic acids, toxins, lipoproteins, and enzymes have been recognised in these nano-sized vesicles that may be involved in bacterial physiology and pathogenesis. The intrinsic adjuvant effect, native immunogenic cargo, sensing by host immune cells, circulation in all body fluids, and comprehensive distribution of antigens introduce EVs as a promising tool for designing novel vaccines, diagnostic biomarkers, and drug delivery systems. Genetic engineering of the EV-producing bacteria and the subsequent production of proper EVs could facilitate the development of the EV-based therapeutic applications. Recently, it was demonstrated that thick-walled mycobacteria release EVs, which contain immunodominant cargos such as lipoglycans and lipoproteins. The present article is a comprehensive review on the recent findings of Mtb EVs biology and the exploitation of EVs for the vaccine technology and diagnostic methods.

Molecular dynamics study of nitrogen diffusion in nanocrystalline iron.

To obtain a deeper understanding of the production of advanced high-nitrogen steels, the diffusivity of nitrogen in bcc iron was investigated at the nanoscale by molecular dynamics (MD) simulation using the modified embedded-atom method (MEAM) interatomic potential. The diffusivity of nitrogen was calculated by mean square displacement (MSD) at different temperatures (773-1473 K) and nitrogen concentrations (0.23, 0.77, and 1.50 wt.%). The results show that the diffusion coefficient increases with temperature and decreases with increasing nitrogen concentration. The temperature dependence of the diffusion coefficient according to the Arrhenius equation was obtained. Activation energies and pre-exponential factors for different nitrogen concentrations were derived from the diffusion coefficient.