Development, production and characterization of SARS-CoV-2 virus-like particles (Coronaviridae: Orthocoronavirinae: Betacoronavirus: Sarbecovirus).

INTRODUCTION: The COVID-19 pandemic caused by SARS-CoV-2 has created serious health problems worldwide. The most effective way to prevent the occurrence of new epidemic outbreaks is vaccination. One of the modern and effective approaches to vaccine development is the use of virus-like particles (VLPs). The aim of the study is to develop a technology for production of VLP based on recombinant SARS-CoV-2 proteins (E, M, N and S) in insect cells. MATERIALS AND METHODS: Synthetic genes encoding coronavirus proteins E, M, N and S were used. VLP with various surface proteins of strains similar to the Wuhan virus, Delta, Alpha and Omicron were developed and cloned into the pFastBac plasmid. The proteins were synthesized in the baculovirus expression system and assembled into VLP in the portable Trichoplusia ni cell. The presence of insertion in the baculovirus genome was determined by PCR. ELISA and immunoblotting were used to study the antigenic activity of VLP. VLP purification was performed by ultracentrifugation using 20% sucrose. Morphology was assessed using electron microscopy and dynamic light scattering. RESULTS: VLPs consisting of recombinant SARS-CoV-2 proteins (S, M, E and N) were obtained and characterized. The specific binding of antigenic determinants in synthesized VLPs with antibodies to SARS-CoV-2 proteins has been demonstrated. The immunogenic properties of VLPs have been studied. CONCLUSION: The production and purification of recombinant VLPs consisting of full-length SARS-CoV-2 proteins with a universal set of surface antigens have been developed and optimized. Self-assembling particles that mimic the coronavirus virion induce a specific immune response against SARS-CoV-2.

Role of the trace amine associated receptor 5 (TAAR5) in the sensorimotor functions.

Classical monoamines are well-known modulators of sensorimotor neural networks. However, the role of trace amines and their receptors in sensorimotor function remains unexplored. Using trace amine-associated receptor 5 knockout (TAAR5-KO) mice, that express beta-galactosidase mapping its localization, we observed TAAR5 expression in the Purkinje cells of the cerebellum and the medial vestibular nucleus, suggesting that TAAR5 might be involved in the vestibular and motor control. Accordingly, in various behavioral tests, TAAR5-KO mice demonstrated lower endurance, but better coordination and balance compared to wild-type controls. Furthermore, we found specific changes in striatal local field potentials and motor cortex electrocorticogram, such as a decrease in delta and an increase in theta oscillations of power spectra, respectively. The obtained data indicate that TAAR5 plays a considerable role in regulation postural stability, muscle force, balance, and motor coordination during active movements, likely via modulation of monoaminergic systems at different levels of sensorimotor control involving critical brain areas such as the brainstem, cerebellum, and forebrain.

Minimal Age-Related Alterations in Behavioral and Hematological Parameters in Trace Amine-Associated Receptor 1 (TAAR1) Knockout Mice.

Since the discovery in 2001, the G protein-coupled trace amine-associated receptor 1 (TAAR1) has become an important focus of research targeted on evaluation of its role in the central nervous system (CNS). Meanwhile, impact of TAAR1 in the peripheral organs is less investigated. Expression of TAAR1 was demonstrated in different peripheral tissues: pancreatic ß-cells, stomach, intestines, white blood cells (WBC), and thyroid. However, the role of TAAR1 in regulation of hematological parameters has not been investigated yet. In this study, we performed analysis of anxiety-related behaviors, a complete blood count (CBC), erythrocyte fragility, as well as FT3/FT4 thyroid hormones levels in adult and middle-aged TAAR1 knockout mice. Complete blood count analysis was performed on a Siemens Advia 2120i hematology analyzer and included more than 35 measured and calculated parameters. Erythrocyte fragility test evaluated spherocytosis pathologies of red blood cells (RBC). No significant alterations in essentially all these parameters were found in mice without TAAR1. However, comparative aging analysis has revealed a decreased neutrophils level in the middle-aged TAAR1 knockout mouse group. Minimal alterations in these parameters observed in TAAR1 knockout mice suggest that future TAAR1-based therapies should exert little hematological effect and thus will likely have a good safety profile.

Characteristics of lipid fractions of larvae of the black soldier fly Hermetia illucens.

The lipid fraction of larvae of the black soldier fly Hermetia illucens was shown to contain lauric acid (38.43 wt %) and its esters, azelaic and sebacic acids, and azelaic acid dibutyl ester. The dominant compound in the group of identified glycerides was lauric acid monoglyceride (0.70 wt %). Glycerides were also represented by triglycerides and diglycerides of lauric acid. Sterols were represented primarily by phytosterols (over 75%), the major of which was alpha-sitosterol (45%). The identified lipid complex composition is apparently determined by the biological characteristics of the fly Hermetia illucens and ensures antibacterial defence of larvae and stability of lipids at changing ambient temperature.

[BLOCKING SENSORIMOTOR DISORDERS IN STRIATIC DOPAMINERGIC DEFICIENT MICE BY THE EXTRACTS OF "REMEDY BEETLE" ALPHITOBIUS DIAPERINUS].

Five groups of C57Bl/6JSto mice (groups 1-5) were tested on a vertical pole 2 weeks since subcutaneous administration of proneurotoxin MPTP (40 mg per kg b. w.) to groups 2-5. Group 1 served as control. It is known that this MPTP dosage forms within 2 weeks sensorimotor disturbances similar to the initial stage of Parkinson's disease. One week before MPTP injection and during all the period after it (2 weeks) animals of groups 3-5 got one of the three extracts of biomass homogenate of the «remedy beetle¼ Alphitobius diaperinus as an additive to their food (8 g per 1 kg of food). The extracts were as follows: water (N 1), water-methanol (N 2) and water-methanol one after solid-phase extraction (N 3). All extracts were immobilized on a phytocarrier. Animals of groups 1 & 2 used ordinary food. Sensorimotor disturbances which could be clearly seen in the 2nd group of mice (MPTP) were absent in group 1 (control) as well as groups 3-5 which received an «antidote¼ with their food as one of the extracts of homogenate of the beetle biomass. The extract N 3 demonstrated maximal efficiency of all.

Features of the Effect of a Complex Probiotic with Bacillus Bacteria and the Larvae of Hermetia illucens Biomass on Mozambique Tilapia (Oreochromis mossambicus O. niloticus) and Russian Sturgeon (Acipenser gueldenstaedti) Fry.

A study of the effect of a complex probiotic preparation with bacteria of the genus Bacillus and a microdose of larvae of the fly Hermetia illucens on the physiological. parameters of fish has been carried out. It was established that introduction of the complex into artificial feeds for tilapia and Russian sturgeon juveniles during their incubation in recirculating aquaculture systems improves the digestion efficiency and increases the rate of fish growth and body mass accumulation. In tilapia, a decrease in the cholesterol level was revealed, while in sturgeon, an increase in the hemoglobin content was found. It was noted that a probiotic with biologically active agents H. illucens had a stimulating effect on the development of sexual gonads in female sturgeon fry, promoted an increase in the ovary mass, and provided for the development of eggs without morphological disturbances.

[Special effects of a complex probiotic containing cellulolytic bacteria Cellulomonas on actively growing rabbits].

It was shown that the association of probiotic bacteria of the genuses Bacillus and Cellulomonas form biolayers on the surface of beet marc particles. The positive effect of a fodder additive that contained the biolayer on the basis of a phytomatrix on the growth and development of young rabbits was shown. Feeding of animals with a mixed fodder that contained 0.1% preparation resulted in stimulation of digestion of all components of the food. Among other components of the mixed fodder, cellulose was digested most effectively. An increase in the biomass of symbiotic bacteria and enzymatic activity in the blindgut chymus was also observed. The positive nitrogen balance demonstrated an increase in the nitrogen content in animals and a decrease of its losses with excretion. The mechanism of response of the rabbit's organism to introduction of the complex probiotic preparation into the digestive tract is discussed.

[Anaerobic solid-phase fermentation of plant substrates by Bacillus subtilis].

Solid-phase growth of Bacillus subtilis 8130 on cellulose-rich plant substrates (presscakes or pulp) under hypoxic conditions was accompanied by cellulose depolymerization, protein hydrolysis, and degradation of other plant components, including some processes of mixed-type carbohydrate fermentation. The bacterial fermentation yielded propionic, butyric, and hexanoic acids and butyric acid derivatives. The bacterial metabolism and fermentation degree can be characterized by the proportions of fatty acids in the reaction mixture. The product of sea buckthorn cake fermentation has a good sorption quality.

[A study of the mechanisms of probiotic effect of Bacillus subtilis 8130 strain].

The wild-type Bacillus subtilis strain 8130 secreted metabolites that stimulated two to three times the growth of the test cultures of lactic acid bacteria. It exhibited endoglucanase activity that depended on the composition of nutrient medium. The addition of the product of two-stage culturing of B. subtilis 8130 to the diet of pigs (0.2% of fodder weight) made it possible to increase the daily weight gain by 19% and decrease the consumption of mixed fodder by 10%. Digestion of protein, fat, and other organic compounds increased by 3-4% and cellulose by 12%. It was shown that B. subtilis 8130 is a probiotic with targeted action stimulating digestion (primarily the digestion of cellulose). The enrichment of a dry-beer pellet with the product of solid-phase fermentation by bacillus (1 x 10(8) cells per gram dry pellet) allowed the pellet to entered into the diet of a calf (6% of the weight of fodder with probiotic), causing additional weight gain by 12% and a 10% economy of fodder consumption.

[A2144G is the main mutation in the 23S rRNA gene of Helicobacter pylori associated with clarithromycin resistance].

To detect point mutations A2115C, A2143G/C, and A2143G in the 23S rRNA gene of Helicobacter pylori associated with resistance of the microorganism to clarithromycin, a new powerful way of analysis was used. This method involved the reaction of minisequencing followed by MALDI-TOF mass spectrometry of reaction products. In ten analyzed clarithromycin-resistant clinical isolates of H. pylori obtained in Russia, the resistance was found to be mediated only by mutation A2144G in the 23S rRNA gene.

[Cellulose decomposition under nitrogen deficiency by bacteria isolated from the intestines of phytophagous vertebrates]. / Rasshcheplenie tsellulozy pri defitsite azota bakteriiami, vydelennymi iz kishechnika rastitel'noiadnykh pozvonochnykh.

A nitrogen-fixing strain identified as Klebsiella pneumonia 402-2 and two endoglucanase-synthesizing Bacillus strains were isolated from the intestines of phytophagous animals. One of the Bacillus strains was identified as Bacillus subtilis GL. Klebsiella pneumoniae 402-2 increased the endoglucanase activity of both Bacillus strains in mixed cultures. The data on the taxonomic position of strains 402-2 and GL and on the nitrogen-fixing capacity of strain 402-2 were confirmed by sequencing and analyzing their 16S rRNA genes and by amplifying the nitrogenase gene nifH.

[Histological and immunohistochemical characteristics of diffuse large cell B-cell lymphomas]. / Gistologicheskaia i immunogistokhimicheskaia kharakteristika diffuznykh krupnokletochnykh B-kletochnykh limfom.

In the REAL classification, diffuse large B-cell non-Hodgkin's lymphomas are grouped together. We investigated histological variants and immunohistochemical profile of diffuse large B-cell lymphomas in 53 patients. Accuracy of the diagnosis was 73.6% without immunohistochemistry. The usefulness of immunophenotyping in making the correct diagnosis depended on a specific histological variant of diffuse large B-cell lymphoma. Variants with polymorphic and anaplastic morphology or massive reactive component have been diagnosed by routine histological methods with poor validity.

[Neurologic complications in otitis and rhinosinusitis]. / Nevrologicheskie oslozheniia pri otitakh i rinosinusitakh.

In 67 patients (32 with otitis and 35 with rhinosinusitis) otorhinosinusogenic complications were detected (cerebral abscesses, meningoencephalitides, cerebral arachnoiditides, encephalopathies, vasoautonomic forms of hypothalamic syndrome). In one patient testaceous pachymeningitis ran uncommon course with a fatal outcome.