The development of new humanized transgenic mouse biomodels with the HLA-A*02:01:01:01 gene requires effective methods for target transgene verification in the animal genome. In the present study, we develop a system for genetic screening of animals based on real-time PCR and using highly specific primers to detect all functionally significant parts of the genetic construct. In addition, the Sanger sequencing method showed the absence of chimerism and complete correspondence between the primary nucleotide sequence of the HLA A*02:01:01:01 transgene and the developed engineered genetic construct and human gene HLA A*02:01:01:01. Based on the results of selection and genetic works with the resulting transgenic animals, three most promising sublines were identified. These lines are currently used for breeding a new line of humanized transgenic mice with the HLA-A*02:01:01:01 gene.

The Western Diet (WD) is a nutritional style characterized by excessive intake of cholesterol, saturated fatty acids and sugars; this nutritional pattern can cause type 2 diabetes, metabolic syndrome and other disorders, particularly during ageing. The search for effective approaches to managing the metabolic syndrome caused by WD seems to be a relevant research task. Unfortunately, this issue has attracted insufficient attention in the current literature. In this work, we use a mouse model of WD to study potential effects of a food supplement based on dicholine succinate (DS) and folic acid (vitamin B₉), which are activators of mitochondrial functions. We study glucose tolerance, parameters of hippocampus-dependent learning and relative gene expression in RT-PCR of molecular markers of negative WD effects in the brain and liver of aging mice housed on WD. Mice C57BL/6 were 12 months old and housed on WD for 3 weeks; some groups received В₉ (5 mg/kg/day) or DS (150 mg/kg/day), or their combination, via water. We carried out food displacement and fear conditioning learning tests followed by RT-PCR of several genes in the liver and brain. We found a decreased glucose tolerance, an elevated speed of pellet displacement and a reduction of freezing time in the fear conditioning test. This may suggest cognitive deficits and impulsivity of mice housed on WD. The administration of DS and B diminished most of these changes. In addition, the increased expression of FASN in the liver points to new mechanisms of negative WD effects during aging. The food supplement based on B₉ and DS normalizes FASN expression and behavior, as well as glucose tolerance in WD-housed mice. Our results open new perspectives for further studies of therapeutic and preventive effects of food supplements on the regulation of metabolic parameters during ageing.

Systemic response to stress, aimed at eliminating or reducing stress, is accompanied by changes in behavioral, vegetative, motor, sensory, cognitive, and other bodily functions. Stress behavior is an integral part of general behavior. In this work, we set out to study the behavioral activity of rats in an open field test following administration of various lithium salts. The conducted experiments showed that a course application of organic lithium salts improves the motivational characteristics of behavior and the appearance of rats, significantly increases the tentative research behavior of experimental animals, and reduces the level of anxiety, fear, and uncertainty, which can be characterized as an adaptive-stimulating effect. At the same time, the drug demonstrates a cumulative effect. The anabolic effect of lithium orotate is particularly pronounced, which makes it possible to increase body weight gains in the setting of stress of various etiologies.

Hormonal preparation of female egg donors is one of the first stages in obtaining genetically modified animals by microinjected zygote transplantation. A large number of embryos synchronized in development can be obtained by a superovulation procedure based on treating female animals with gonadotropins. As a rule, drugs from different manufacturers differ markedly in terms of their efficacy. In the context of sanctions imposed on the Russian Federation, attention should be paid to domestic hormonal drugs, thus substantiating their objective assessment. It was shown that different batches of the FSH-SUPER drug, used mainly for hormonal treatment of farm animals, differed in stimulating effect on superovulation in female mice. At the same time, the efficacy of this drug surpassed that of Follimag (Mosagrogen, Russia).

N-terminal acetyltransferases (NATs) of bacteria acetylate the alpha-amino group in amino acids and proteins, participate in the biosynthesis of lantibiotics, and inactivate a number of antibiotics. NATs are used in biotechnology for targeted acetylation of recombinant proteins and peptides. In this regard, the search for NATs that differ in terms of substrate specificity and are also capable of functioning in the reaction at elevated temperatures, a wide pH range, etc., seems relevant. In this work, we develop specific characteristics and a search algorithm for the identification of N-terminal acetyltransferases using the Thermus thermophilus thermophilic bacterium as an example. Out of 14 Abs annotated in the genome, we selected six «putative» NATs. Some of the genes encoding the selected NATs were successfully cloned, generated, and purified from E. coli cells. The specific enzymatic activity of a number of enzymes was confirmed.

Cleavage of the N-terminal initiating methionine (Met1) is a critical coand post-translational modification affecting 50–70% of cellular proteins. During the production of recombinant proteins in the heterologous system of E. coli expression, Met1 cleavage often fails to occur, which leads to heterogeneity of the preparations obtained, changes in their activity and stability. This problem can be solved by treating recombinant proteins in vitro with a specific enzyme, methionine aminopeptidase (MAP). Currently available MAPs exhibit limited specificities and reaction conditions. We cloned a MAP from a hyperthermophilic bacterium, developed a method for enzyme purification, and studied a number of physicochemical properties. The new MAP enzyme is resistant to elevated temperatures. The MAP maintains a stable native state in a pH range from 3 to 11 units. The novel MAP enzyme can be used to remove N-terminal Met1 from recombinant proteins in vitro over a wide pH range and at elevated temperatures.

This article describes the process of developing a tissue-engineered structure that meets the biocompatibility and biodegradation parameters necessary for replacing cartilage tissue defects. The study was carried out using 3D bioprinting technology, which represents a promising research direction in the biomedical field. It is known that, due to the specifics of its structure, cartilage tissue is not capable of complete regeneration of damage. The methods currently used for treating arthrosis are associated with a number of limitations and disadvantages, which makes research aimed at developing alternative methods for arthrosis treatment particularly relevant. The development of tissue-engineered structures by 3D bioprinting requires the materials not only certified for medical use but also exhibiting biocompatibility and biodegradation properties. Polylactide (PLA) and sodium alginate satisfy the above requirements; moreover, their availability and economic affordability make them one of the most popular materials for 3D bioprinting.

We studied the effect of infrared radiation (IR) course exposure on pathological changes in the tissues of C57BL/KsJYLepr^db/+ (db/db) mutant mice with type 2 diabetes mellitus using a LASMA ST device. This apparatus allows simultaneous non-invasive monitoring of blood and lymph microcirculation, as well as determining the activity of mitochondrial oxidative coenzymes in tissues before and after IR therapy. IR waves (690 nm) were found to have a positive therapeutic effect on the functional state of db/db mice. This included a rapid healing of skin wounds caused by maceration and prolonged normalization of oxidative metabolism parameters (NADH, FAD, POM) in body tissues. IR therapy has a cumulative effect, leading to a 2.3–2.5-fold increase in life expectancy and life quality, compared to the control group.

We study the induction patterns of the solid form of Ehrlich ascitic carcinoma (EAC) in mice in the short(1 month) and long-term (18 months) periods after inoculation of ascitic cells irradiated ex vivo with a proton beam at doses of 30, 60, 90, 120, and 150 Gr. The growth rate of solid tumors after inoculation of irradiated cells ex vivo coincided with the growth of tumors in the control group. The frequency of tumor induction in mice transplanted with EAC cells was dose-dependent and decreased from 80% to 0% along with an increase in radiation dose. Thus, using the ex vivo model of tumor cell irradiation followed by in vivo grafting allowed us to determine the parameters of the antitumor effect of new proton therapy regimens.

A scheme for correcting intestinal motor disorders and microbiocenosis is proposed. Experiments on rats included a sequential jejunum administration of pharmaconutrient enteral nutrition (Intestamine, containing glutamine and tributyrin) and inulin, a prebiotic preparation. The application of this scheme in the postoperative period in the setting of endotoxemia prevents excessive bacterial growth in the jejunum, stimulates the appearance of propulsive peristalsis, eliminates pathological motility patterns and promotes early recovery of coordinated peristalsis of the small intestine.

In this work, we studied the effect of heparin on the binding process of VEGF-A165, the most common and important growth factor, and the monoclonal antibody bevacizumab by bio-layer interferometry. A complex of the full-length bevacizumab antibody with VEGF-A165 was modeled. The data obtained can be used in therapy with this antibody, as well as in the development of other therapeutic antibodies with maximum target specificity under various conditions.

Daphnia magna Straus was used as an experimental zoohydrobiont model for primary screening of compounds of the valproic acid aminoether group with anticonvulsant and cholinoblocking activity. On this basis, the most effective compounds for further trials in warm-blooded animals were selected. The selection criterion included their minimum effective concentration for the prevention of atypical motor hyperactivity induced by phenylcarbamate.

Despite the diversity of modern treatment methods, infertility rates remain high both in Russian and globally. In the present experimental study, we study the effect of a nanopreparation based on a selenoxanthene derivative and cyclodextrin on the reproduction of white outbred rats (females) after the administration of mifepristone. Selenoxanthene at a dose of 0.35 mg/kg and 0.7 mg/kg intragastrically for 5 days increased the reproductive activity of animals.

In this work, we investigate the anti-inflammatory effect of the HLDF-6-H peptide (Thr-Gly-Glu-HseHis-Arg) in a model of the presymptomatic stage of Parkinson’s disease (PD). This peptide is a bioactive fragment of the HLDF leukocyte differentiation factor. The presymptomatic stage of PD was modelled by introducing moderate doses of MPTP toxin to C57Bl/6 mice. HLDF-6-H was administered intranasally at a dose of 300 μg/kg daily for three weeks. The severity of depression was determined by the time of immobilization in a Porsolt test. The serum levels of 10 steroids were determined using MS analysis. The activity of inflammatory markers leukocyte elastase (LE) and α1-proteinase inhibitor (α1-PI) was determined using kinetic methods. The development of an inflammatory process under the influence of MPTP is evidenced by an increase in the activity of α1-PI; the anti-inflammatory effect of the peptide is manifested in a decreased activity of LE and α1-PI. At the same time, the immobilization time of mice in a Porsolt test positively correlated with α1-PI and LE, which may indicate the role of inflammation in the development of depression. A positive correlation between estradiol and LE was found in the general sample of mice. In the group of mice treated with the peptide, α1-PI correlated positively with corticosterone and negatively with estradiol and androstenedione. It can be assumed that the anti-inflammatory effect of the peptide is associated with the regulation of corticosteroids and sex hormones. Hence, the applied experimental model confirmed the development of a depressive-like syndrome and neuroendocrine disorders in the presymptomatic stage of PD. The long-term use of HLDF-6-H has antidepressant and anti-inflammatory effects.

We present the results of experimental studies into the pharmacological safety of a pharmaceutical substance 4-[(3-oxo-3-ethoxypropanoyl)amino]benzoic acid (Etmaben) under a single intragastric administration at various experimental doses (60–3000 mg/kg) to female laboratory outbred rats. A clinical examination of the experimental animals was carried out in order to identify signs of intoxication. To that end, the level of food and water consumption was assessed and biochemical blood tests were performed. In addition, the state of the central nervous and cardiovascular systems, physical endurance, and respiratory rate were assessed. The tested pharmaceutical substance in the studied doses after a single intragastric administration to female rats had no negative effect on the behavior, blood parameters, the state of the central nervous system and the cardiovascular system of experimental animals.

A comparative evaluation of the effectiveness of dry alcoholic extracts of Dioscorea from pharmacopoeial raw materials and cell cultures was carried out on a model of combined exposure to hypoxic and temperature factors and swimming of laboratory animals (mice) with a load of 5% of body weight. Dioscorea preparations had a pronounced thermoprotective effect and increased physical performance; however, the aftereffect was confirmed only for the cell culture extract.

This paper presents a technology for creating customized porous titanium implants with bioactive coatings, manufactured using additive technologies. The stages of creating an implant include obtaining primary data from the bone defect area using computed tomography; 3D modeling of the defect area and the corresponding implant; production of a customized implant from titanium alloys using selective laser fusion technology; application of bioactive coatings. The as-created customized implant can have several functional structures. Samples of titanium implants with bioactive coatings were subjected to extensive testing. Mathematical modeling and experiments were used to verify the correspondence of the mechanical properties of the developed structures to natural bone tissue. In vitro tests of the studied samples showed the absence of acute toxicity along with high levels of biocompatibility. In vivo tests of the studied samples on Soviet chinchilla rabbits and Anubis baboon monkeys approved by local ethical committees showed their adequate biomechanical and high osteoinductive properties. The successful results of preclinical studies, as well as toxicological and technical tests in certified laboratories, made it possible to create a registration dossier for state registration of customized porous titanium implants with bioactive coatings, manufactured using additive technologies.

A new derivative of dimethylaminoethanol, butanedioic and trans-butenedioic acids (laboratory code ADK-17) was synthesized at the Organic Chemistry Department of the Saint Petersburg State Chemical and Pharmaceutical University (SPSPU). This promising nootropic and anti-asthmatic agent is planned for use in an oral dosage form. The aim was to calculate the pharmacokinetic parameters of the synthesized compound and to determine its possible biotransformation pathways. Rabbits of the Soviet Chinchilla breed were used as a test system in the preclinical study of the compound pharmacokinetics. In accordance with the designed schedule, blood was taken from the marginal vein of the ear, and urine samples were taken. Quantitative determination in biological media was carried out using an Ultimate 3000 liquid chromatograph with a Q-Exactive mass-selective detector with electrospray ionization in accordance with OFS 1.2.1.100015 (Chromatography) of the State Pharmacopoeia by ultra-performance liquid chromatography–mass spectrometry (UPLC-MS). To study the pharmacokinetics of the ADK-17 preparation, a measurement procedure was developed with a detection limit in biological media ranging from 10-6 to 10^-3 mg/ml. The determined harmacokinetic parameters show that the absorption of ADK-17 into the systemic circulation from the gastrointestinal tract occurs at an average rate, with its maximum concentration in blood plasma during the oral route of administration being observed between 60–90 min. The elimination half-life from the systemic circulation comprises 4.3 h. Urinary excretion in the first 4 h proceeds at a high rate; however, the compound is detected in the urine within 3 days and in the blood within 2 days from the moment of administration.

Ligands of human serum albumin (HSA) are capable of modulating its interaction with β-amyloid peptide (Aβ), which is a key factor in the pathogenesis of Alzheimer’s disease (AD). L-thyroxine (L-Tr), a natural HSA ligand, is associated with the pathogenesis of AD according to epidemiological and animal model studies. In this work, we studied the kinetics of Aβ fibril formation in the presence of L-Tr and HSA using a fluorescent test with thioflavin T. L-Tr had no significant effect on the inhibitory effect of HSA on fibril growth. At the same time, L-Tr itself had an inhibitory effect similar to that of HSA. Our data can partially explain the relationship between AD and thyroid pathologies.

Probiotics are microbial preparations, which have a positive effect on the composition of the intestinal microflora and the immune system of animals. These preparations are widely used for correcting and preventing violations of the microflora of the gastrointestinal tract. Lactobacilli play a special role in the formation of normal microflora. They exhibit a pronounced antagonistic activity against pathogenic and opportunistic microorganisms, as well as possess a number of other positive effects. This study is aimed at determining the effect of new probiotic strains of L. reuteri on the indicators of nonspecific resistance, hematological and biochemical parameters of laboratory rabbits. The studied strains were found to have a pronounced positive effect on the indicators of nonspecific resistance. Thus, compared to the control, the indicators of phagocytic activity and bactericidal activity in the experimental group were by 22% and 32% higher, respectively. The content of hemoglobin, erythrocytes and total protein in the blood serum also increased.

Research into the consequences of human exposure to heavy metals acquires particular significance in the context of life and working age prolongation. Such consequences can be manifested either in the form of selective damage to any organ, organ system or function, or in the form of a general violation of the functions of a number of systems and organs and the organism as a whole, in the form of “disintegration of functions”. In this work, we set out to study the neuroprotective effect of etomerzol fumarate after a subchronic low-dose administration of mercury acetate to Wistar rats. After an intragastric administration of an aqueous solution of mercury acetate at a dose of 4 mg/kg for 30 days, a violation of the learning process was identified in an active avoidance by swimming test. The use of the test substance at a dose of 12.5 mg/kg for 14 days had a positive effect on the long-term memory of animals, contributing to the preservation and reproduction of mercury information received.

Aquagenic urticaria is one of the rarest forms of allergic reaction in the body. Aquagenic urticaria is clinically manifested by itching and urticarial rashes 5–10 min after contact with water. The signs disappear within 30–60 min. The standard therapy regimen includes the administration of second-generation H1-histamine receptor blockers and barrier creams with ceramides. In recent years, the use of monoclonal antibody therapy (omalizumab) has been considered in cases where antihistamines are ineffective.