Russian Ministry of Health

Protease S (PrtS) from Photorhabdus laumondii belongs to the group of protealysin-like proteases (PLPs), which are understudied factors thought to play a role in the interaction of bacteria with other organisms. Since P. laumondii is an insect pathogen and a nematode symbiont, the analysis of the biological functions of PLPs using the PrtS model provides novel data on diverse types of interactions between bacteria and hosts.

Recombinant PrtS was produced in Escherichia coli. Efficient inhibition of PrtS activity by photorin, a recently discovered emfourin-like protein inhibitor from P. laumondii, was demonstrated. The Galleria mellonella was utilized to examine the insect toxicity of PrtS and the impact of PrtS on hemolymph proteins in vitro. The insect toxicity of PrtS is reduced compared to protease homologues from non-pathogenic bacteria and is likely not essential for the infection process. However, using proteomic analysis, potential PrtS targets have been identified in the hemolymph.

The spectrum of identified proteins indicates that the function of PrtS is to modulate the insect immune response. Further studies of PLPs' biological role in the PrtS and P. laumondii model must clarify the details of PrtS interaction with the insect immune system during bacterial infection.

Recently, we have described the first supermolecular nanoentities of vitamin B₁₂ derivative, viz. monocyano form of heptabutyl cobyrinate, unique nanoparticles with strong noncovalent intermolecular interactions, emerging optical and catalytic properties. Their nearest analogue, heptamethyl cobyrinate (ACCby), exhibits bioactivity. Here, we demonstrate the first example of the formation of nanoparticles of this nucleotide-free analogue of vitamin B₁₂ in protein nanocarriers and neuroprotective activity in vivo of the own nanoform of the drug. The preparation and characterization of nanocarriers based on bovine serum albumin (BSA) loaded with vitamin B₁₂ (viz. cyano- and aquacobalamins) and ACCby were performed. Nucleotide-free analogue of vitamin B₁₂ is tightly retained by the protein structure and exists in an incorporated state in the form of nanoparticles. The effect of encapsulated drugs on the character and severity of primary generalized seizures in rats induced by the pharmacotoxicant thiosemicarbazide was studied. Cyanocobalamin and ACCby exhibited a neuroprotective effect. The best influence of the encapsulation on the effectiveness of the drugs was achieved in the case of AСCby, whose bioavailability as a neuroprotector did not change upon introduction in BSA particles, i.e., 33 % of surviving animals were observed upon ACCby administration in free form and in encapsulated state. No surviving rats were observed without the administration of drugs. Thus, BSA nanocarriers loaded by nanoparticles of nucleotide-free analogues of vitamin B_12, including hydrophobic ones, can be recommended for neuroprotection and targeted delivery.

Bioengineered vascular grafts (VGs) have emerged as a promising alternative to the treatment of damaged or occlusive vessels. It is thought that polyurethane (PU)-based scaffolds possess suitable hemocompatibility and biomechanics comparable to those of normal blood vessels. In this study, we investigated the properties of electrospun scaffolds comprising various blends of biostable polycarbonate-based PU (Carbothane™ 3575A) and gelatin. Scaffolds were characterized by scanning electron microscopy, infra-red spectroscopy, small-angle x-ray scattering, stress-loading tests, and interactions with primary human cells and blood. Data from in vitro experiments demonstrated that a scaffold produced from a blend of 5% Carbothane™ 3575A and 10% gelatin has proven to be a suitable material for fabricating a small-diameter VG. A comparative in vivo study of such VGs and expanded polytetrafluoroethylene (ePTFE) grafts implanted in the abdominal aorta of Wistar rats was performed. The data of intravital study and histological examination indicated that Carbothane-based electrospun grafts outclass ePTFE grafts and represent a promising device for preclinical studies to satisfy vascular surgery needs.