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In this study, Methylophilus quaylei proved to possess the ability to reduce AgNO3 and produce silver nanoparticles (AgNPs) under specific conditions via bacterial cells and cell-free supernatant (CFS). The obtained AgNPs via bacterial cells were examined by the transmission of electron microscopy (TEM) consistent with X-ray microanalysis and for AgNPs via CFS by visible (UV–Vis) absorption spectroscopy, scanning electron microscopy (SEM), and dynamic light scattering (DLS). The purpose of this study was to evaluate the antibiofilm effect of AgNPs. The effectiveness of the minimum inhibitory concentration (MIC) of AgNPs in supplementation with CFS was determined on established biofilms on polypropylene by the number of colony forming unit (CFU). These nanoparticles exhibit an antibiofilm effect. CFS was tested individually and in supplementation with the MIC AgNPs obtained on established biofilms of M. quaylei MT and M. quaylei SM. The CFS individually decreased established biofilm for both strains. Further, the obtained MIC of AgNPs in combination with CFS exhibited significant antibiofilm effect against established biofilms for both strains by the destruction of biofilm architecture and detecting cell morphological abnormalities that were observed by SEM. These results showed that the obtained AgNPs had a strong antibiofilm effect against the examined strains.

Along with their excellent safety profiles, subunit vaccines are typically characterized by much weaker immunogenicity and protection efficacy compared to whole-pathogen vaccines. Here, we present an approach aimed at bridging this disadvantage that is based on synergistic collaboration between pattern-recognition receptors (PRRs) belonging to different families. We prepared a model subunit vaccine formulation using an influenza hemagglutinin antigen incorporated into poly-(D,L-lactic-co-glycolic acid) (PLGA) nanoparticles adjuvanted with monophosphoryl lipid A (TLR4 agonist) and muramyl dipeptide (NOD2 agonist). The efficacy studies were conducted in comparison to control vaccine formulations containing individual PRR agonists. We show that the complex adjuvant based on TLR4 and NOD2 agonists potentiates proinflammatory cell responses (measured by activity of transcription factors and cytokine production both in vitro and in vivo) and enhances the phagocytosis of vaccine particles up to comparable levels of influenza virus uptake. Finally, mice immunized with vaccine nanoparticles containing both PRR agonists exhibited enhanced humoral (IgG, hemagglutination-inhibition antibody titers) and cellular (percentage of proliferating CD4+ T-cells, production of IFNɣ) immunity, leading to increased resistance to lethal influenza challenge. These results support the idea that complex adjuvants stimulating different PRRs may present a better alternative to individual PAMP-based adjuvants and could further narrow the gap between the efficacy of subunit versus whole-pathogen vaccines.

The use of recombinant endolysins is a promising approach for antimicrobial therapy capable of counteracting the spread of antibiotic-resistant strains. To obtain the necessary biotechnological product, diverse peptide tags are often fused to the endolysin sequence to simplify enzyme purification, improve its ability to permeabilize the bacterial outer membrane, etc. We compared the effects of two different types of protein modifications on endolysin LysECD7 bactericidal activity in vitro and demonstrated that it is significantly modulated by specific permeabilizing antimicrobial peptides, as well as by widely used histidine tags. Thus, the tags selected for the study of endolysins and during the development of biotechnological preparations should be used with the appropriate precautions to minimize false conclusions about endolysin properties. Further, modifications of LysECD7 allowed us to obtain a lytic enzyme that was largely devoid of the disadvantages of the native protein and was active over the spectra of conditions, with high in vitro bactericidal activity not only against Gram-negative, but also against Gram-positive, bacteria. This opens up the possibility of developing effective antimicrobials based on N-terminus sheep myeloid peptide of 29 amino acids (SMAP)-modified LysECD7 that can be highly active not only during topical treatment but also for systemic applications in the bloodstream and tissues.

The aim of this work was to compare mesoporous carriers based on silica and magnesium aluminosilicate in the amorphous solid dispersion production. Darunavir has been selected as an active pharmaceutical ingredient that is classified as a Class 2 BCS substance and exists in two commercially available forms: crystalline ethanolate and amorphous. In the course of the study, the conditions for the preparation of amorphous samples with the selected carriers were evaluated within the framework of the most common methods for obtaining solid dispersions - hot-melt extrusion, solvent wetting, and spray drying. It was determined that the obtained dispersion properties almost completely repeat the properties of the corresponding carriers. The resulting dispersions were examined in a dissolution test and the best ones was used to formulate tablets, which were studied in an in vitro dissolution test with the original Prezista. The proposed tablet formulation showed improved dissolution compared to the original one. It was also found that silica supports have a greater positive contribution to darunavir dissolution - both ethanolate or amorphous forms.

Chromobacterium species are common in tropical and subtropical zones in environmental samples according to numerous studies. Here, we describe an environmental case of resident Chromobacterium vaccinii in biofilms associated with Carex spp. roots in Moscow region, Russia (warm-summer humid continental climate zone). We performed broad characterization of individual properties as well as surrounding context for better understanding of the premise of C. vaccinii survival during the winter season. Genome properties of isolated strains propose some insights into adaptation to habit and biofilm mode of life, including social cheaters carrying &Delta, luxR mutation. Isolated C. vaccinii differs from previously described strains in some biochemical properties and some basic characteristics like fatty acid composition as well as unique genome features. Despite potential to modulate membrane fluidity and presence of several genes responsible for cold shock response, isolated C. vaccinii did not survive during exposure to 4 &deg, C, while in the complex biofilm sample, it was safely preserved for at least half a year in vitro at 4 &deg, C. The surrounding bacterial community within the same biofilm with C. vaccinii represented a series of psychrophilic bacterial species, which may share resistance to low temperatures with other species within biofilm and provide C. vaccinii an opportunity to survive during the cold winter season.

This study shows the importance of electron microscopy in the analysis of the interaction of microorganisms (Staphylococcus aureus) with polymeric (polyurethane) dental prostheses. Starting from the biofilm formation and the biodestruction of the plastic material resulting in the production of polyurethane nanoparticles, the focus is on the bacterial secretion of membrane vesicles (in the range of 20-50 nm) loaded with plastic nanoparticles (from 2-3 to 10 nm) and on the toxicological threat that these delivery devices represent when interacting with host cells. The nanoparticles deliverance led by the bacterial infections dynamics opens new ways to the possibility of delivering drugs to selected cells.

This study shows that submicron/nanoparticles found in bacterial cells (S. aureus) incubated with polyurethane (a material commonly used for prostheses in odontostomatology) are a consequence of biodestruction. The presence of polyurethane nanoparticles into bacterial vesicles suggests that the internalization process occurs through endocytosis. TEM and FIB/SEM are a suitable set of correlated instruments and techniques for this multi facet investigation: polyurethane particles influence the properties of S. aureus from the morpho-functional standpoint that may have undesirable effects on the human body. S. aureus and C. albicans are symbiotic microorganisms; it was observed that C. albicans has a similar interaction with polyurethane and an increment of the biodestruction capacity is expected by its mutual work with S. aureus.