Creation of humanized genetically modified animals for biomedical research is an important direction in the activity of the Scientific Center of Biomedical Technologies of the Federal Medical and Biological Agency (FMBA) of Russia. Since 2013, the Center has obtained more than 20 lines of transgenic, knockout, and transgenically-knockout mouse biomodels using classical transgenesis and the plasmid version of the CRISPR/Cas9 technology. The main specialized lines involve humanized transgenic mouse biomodels carrying alleles HLA class I — HLA-В*, HLA-В**, HLA-В***, HLA-С*, HLA-B*07:02, HLA-С*07:02, HLA-A*02:01, HLA-В*** КО, HLA-A** КО, HLA-B**** КО, HLA-С** КО, as well as NAT1, NAT2, АСЕ2 hom, hACE2-Tom (HDRKI), SMN2 (S2), PrPSc, АСЕ2 hom / HLA-C**, β2m mus KO, PrP КО, STE24 КО, SMN (S6) КО, SMN2 КО, PrP КО / PrPSc, NAT1 KO, NAT2 KO. In this article, we review the genetic constructs and methods used to produce genetically modified animals. The article summarizes the long-term experience of using the plasmid variant and discusses the need to switch to the CRISPR/Cas9 ribonucleoprotein complex in the view of its high efficiency and specificity, high frequency of targeted modifications, the ability to use multiple gRNAs, and the insertion of a single copy of the HDR-DNA template (transgene). We also explore the possibility of replacing imported reagents and kits with domestic products.

The search for effective therapies against oncological diseases remains one of the most acute problems in modern healthcare. The side effects of chemotherapy drugs often reduce immunity and lead to multiple complications. In this work, we aimed to evaluate the protective properties of Prunella grandiflora L. against the toxic and genotoxic effects of etoposide using a Drosophila model with mutations in the dFOXO gene. Oregon-R and Foxo25 fly lines were cultured on a medium with the addition of etoposide at a concentration of 0.04 μg/ml and 10% alcoholic extract of P. grandiflora, as well as with the combined introduction of these components. The experiment established no toxic effect of P. grandiflora and its antigenotoxic, geroprotective, and neuroprotective properties in relation to the effects of the antitumor drug etoposide.

High-frequency components of brain activity (γ-rhythms, over 30 Hz) are the key markers of cognitive processes. Their study is relevant for diagnostics, treatment, and prevention of neurodegenerative diseases and obsessive-compulsive disorders, including schizophrenia, epilepsy, Alzheimer's disease, etc., as well as for the development and screening of drug effects. The lack of serially-produced equipment with the technical parameters required for research and ready-made invasive systems with the capacity to analyze the high-frequency range of brain activity, as well as the current geopolitical situation, prompted us to create a domestic biopotential amplifier.

This study presents the results of an investigation into minor biologically active compounds in extracts of Siberian musk deer (Moschus moschiferus) musk using high-performance liquid chromatography coupled with high-resolution mass spectrometry. A qualitative and semi-quantitative analysis was performed on various components, including steroids, terpenoids, peptides, and low-molecular-weight organic compounds with potential pharmacological activity. Several compounds, previously unidentified in this biological material, were detected, enhancing our understanding of the chemical composition of musk deer musk and its potential applications in biomedicine.

The conducted review of scientific literature has established the presence of one species of trematodes, three species of cestodes, nine species of nematodes, three species of protozoa, and two species of ectoparasites in Siberian musk deer in the Altai Republic and the Altai Krai. Other ungulates in the Altai Republic exhibit such pathogens as Fasciola hepatica, paramphystomatids, cysticerci (larvae), and echinococcus (larvae), which can potentially infect musk deer. The causative agents of blood-parasitic diseases in musk deer have not been registered so far. However, the presence of ixode ticks that carry vector-borne diseases (Dermacentor pictus, D. marginatus, D. silvarum, D. nuttali, Ixodes persulcatus, Haemaphysalis concinna) indicates the possibility of their occurrence in musk deer. The conducted analysis of literature data and veterinary reports has identified a list of pathogens that are potentially dangerous to musk deer in the Altai Republic. The study has revealed gaps in the knowledge about the parasite fauna of the Siberian musk deer.

The use of Lactoferrin (Lf) for treating stages I and II of type 2 diabetes mellitus (DM2) initially stabilized the glucose level and weight of db/db mice, followed by normalization of these parameters after the drug discontinuation. By the end of Lf usage at stage III of DM2, these parameters in the animals sharply decreased, leading to their rapid death. In all experiments, under Lf treatment at stages I, II, and III of DM2, the oxidative metabolism index (OMI) was initially decreasing, followed by its growth to higher than the initial level at stages I and II of DM2 after the drug discontinuation, and was sharply decreasing again at stage III. The corresponding stress-adaptive changes were also revealed for blood cells and bone marrow, the development of which was carried out with the participation of programmed cell death mechanisms. Lf used to correct clinical manifestations of DM2 acts as a powerful adaptogen. It is capable of exerting an adjuvant effect on the metabolism and state of blood and bone marrow cells, but only at early stages with preserved adaptive reserves in the body.

Neurotoxicity is a manifestation of intoxication with lead compounds. When developing new drugs for the treatment of lead poisoning, modern methods for assessing the neurotoxic effect of such compounds are of great importance. In view of this, we aimed to study long-term manifestations of lead neurotoxicity using a computerized test system for quantitative assessment of the startle reflex in laboratory animals – the TSE Startle Response System (“TSE”, Germany). Outbred male rats weighing 180–220 g were selected as a test system. The animals were randomized into control and experimental groups. The experimental group received a single intraperitoneal injection of lead acetate at a dose of 300 mg/kg. In the surviving animals, sensory responses were assessed on days 3 and 29 using the neurophysiological test of response to a sudden stimulus, the Startle Response System. In both the shortand long-term periods of acute lead intoxication, a decrease in responses to light, noise, and electrical pain stimuli was noted, which may be a consequence of lead accumulation in brain tissue.

The development of biologically active drugs based on genetic engineering technologies is a promising area in the field of creating vaccines for livestock, in particular, to increase meat productivity in sheep farming. Myostatin is a negative regulator of skeletal muscle growth. Blocking the action of endogenous protein by immunizing animals with recombinant myostatin leads to the development of a specific antibody response and a gain in muscle mass. The effect of three-fold vaccination with recombinant myostatin on the weight gain of lambs (rams and sheep) during the fattening period under pasture–corral conditions was studied. It was shown that the weight gain of immunized rams and sheep during the period of 4.5 months was higher than that in the control group animals by an average of 4 and 2 kg, respectively. The absence of mortality in the experimental group was noted, suggesting a positive effect of E. coli impurity proteins in the recombinant vaccine.

Acute intoxications with pesticides, herbicides, and insecticides based on cholinesterase inhibitors account for up to 50% of hospitalizations due to toxic poisoning, leading to either long-term incapacitation or disability. These compounds predominantly cause dysfunction of the cholinergic mediator system, while exhibiting non-receptor effects. The consequences of severe carbamate poisoning can result in pathological conditions that occur in the long-term period of intoxication, including psycho-organic syndrome, asthenic conditions, neuropathies. The current literature lacks data on preventive or therapeutic drugs to prevent the occurrence of such conditions. The development of effective means of pharmacological correction requires adequate biomodels for studying remote effects of carbamate poisoning.

In experiments on 20 rats, biochemical markers of organ dysfunction were assessed 6 and 24 h after modeling internal intestinal fistulas. It was found that biochemical markers of organ dysfunction allow early detection of impaired protein-synthetic function of the liver, development of cytolysis and cholestasis syndrome, and impaired renal function after modeling internal intestinal fistulas.

In experiments on 55 rats, the effect of body temperature during acute blood loss on the locomotor and exploratory activity of animals on days 3 and 7 of the posthemorrhagic period was assessed. A decrease in exploratory and locomotor activity in animals that had undergone acute blood loss under normothermia was detected. Acute massive blood loss performed under hypothermia does not affect exploratory and locomotor activity, which may indicate neuroprotective mechanisms of natural cooling.

This article presents a comparative description of typical electromyographic patterns of neuromuscular transmission disturbances due to effects of various cholinergic agents. The results obtained can be used when modeling pathologies of the peripheral nervous system and conducting preclinical studies in the field of neuropharmacology and neurotoxicology.

Uridine is a precursor of the metabolic activator of the mitochondrial potassium channel leading to tissue protection from hypoxic damage under oxidative stress. In this research, we studied its effects in rats with lactacystin (LC)-induced Parkinson's disease (PD). The biological effect was recorded by the cytobiochemical method for detecting the physiological regulation of enzyme activity in lymphocytes on a blood smear. The activity of succinate dehydrogenase (SDH), an enzyme of mitochondrial energy supply, and lactate dehydrogenase (LDH), a glycolytic enzyme, was measured. Uridine at a dose of 30 mg/kg for 28 days in the modelled preclinical stage (MPS) of PD causes a decrease in hyperactive SDH to the control level. At the same time, in the modelled clinical stage (MCS), uridine exhibited no such effects. Regarding the LDH glycolysis enzyme, the modelled preclinical stage of PD was characterized by a twofold increase in the activity of this enzyme, with uridine having only a slight effect. In the clinical stage of PD, lactocystin also increased the activity of SDH, with virtually no effect on the activity of LDH, while uridine increased the activity of both enzymes. The revealed protective effect of uridine in MPS in rats is explained by the restoration of early pathological disorders of mitochondria in lymphocytes, with an increase in glycolysis over respiration. This effect is not detected in the MCS, where, apparently, irreversible mitochondrial pathology develops.

This work is aimed at improving the quality of biological modeling of pathological conditions in laboratory animals used for the purposes of developing new medicines. The methodological features of the development, verification, and standardization of new biomodels of pathological conditions for relevant and alternative biomodeling, ensuring their compliance with the requirements of evidence-based medicine, are considered. The development of new models should be based on pathogenetically inducing pathological effects similar to human and animal biomodels. The optimality of the model should be studied at several levels of inducing effects, and the research methods should allow quantifying the severity of the modeled pathological condition. It is shown that the verification of new biological models can be implemented in the process of their pharmacological validation using at least two drugs with a known mechanism of action, exhibiting both activating and suppressing pathogenetically determined effects on the severity of the studied pathological condition. The criterion for a sufficient degree of standardization of the model can be high indicators of intra-laboratory and precision convergence of modeling results.

Secondary metabolites of essential oil crops are capable of exhibiting antibacterial, anti-inflammatory, and immunostimulatory properties in human and animal organisms. In this study, we investigate the immunomodulatory effect of coriander and fennel essential oils using swine models as those most similar to the physiological state of humans. The immune status was assessed by determining the expression of IL-6 and IL-8 genes in swine blood leucocytes. The results established a significant decrease in the expression of cytokines in the experimental groups compared with the control group: IL-6 and IL-8 by 4.69 and 19.31 times in group 1, and by 9.15 and 3.54 times in group 2. The essential oils under investigation were established to demonstrate immunomodulatory effects, allowing animals and humans to respond more plasticity to external factors of various genesis.

In this research, we develop and study an extemporaneous (main) form of 20-hydroxyecdysone, along with drafting of prescription forms. Preclinical studies were conducted using 20-hydroxyecdysone and its extemporaneous form. The latter proved its harmlessness (the drug belongs to class VI of relatively harmless substances according to Hodge and to class V of toxicity in accordance with GOST 32644-2014) and efficacy in a model of experimental hyperglycemia. The developed extemporaneous form of 20-hydroxyecdysone was studied in the form of a nanostructured clathrate complex with arabinogalactan with improved pharmaceutical and pharmacological properties. The nanostructured clathrate complex of 20-hydroxyecdysone with arabinogalactan at a mass ratio of 1:10 was obtained as a finely-dispersed white powder with a light-yellow tint and the average particle size of 35.3 nm. In terms of bioavailability, the extemporaneous form of 20-hydroxyecdysone exceeds the similar value of the original compound by 1.91 times. 20-hydroxyecdysone is involved in the phosphatidylinositol-3-kinase signaling pathway of activation of serine-threonine protein kinase B. In the activated form, this kinase phosphorylates various protein substrates in the cytosol, thereby modulating their functions and playing a central role in various cellular processes. The extemporaneous form of 20-hydroxyecdysone allows its dose to be reduced, thus ensuring high compliance with the drug. Low doses of the drug minimize the probability of side effects and their rapid elimination in case of their emergence. Instructions for the medical use of the developed extemporaneous form of 20-hydroxyecdysone are provided, and prescription forms are developed. Based on the results obtained, the extemporaneous form of 20-hydroxyecdysone can be recommended for use as independent therapy or in combination with other metabolic agents. The established physiological efficacy and safety render the developed extemporaneous form of 20-hydroxyecdysone a metabolic agent for practical use.

In this study, we evaluated the ability of three new chromone-containing allylmorpholines (33a, 33b, and 9a) to exhibit hypothermic effects in rats after a single injection. The temperature was measured rectally before the injection and after 20, 40, and 60 min. It was found that compound 33a at doses of 50, 100, and 150 mg/kg, as well as compound 33b at a dose of 60 mg/kg, were able to decrease body temperature in rats relative to the control group at the same time point. No such effects were detected with compound 9a. These data may indicate the presence of antipsychotic action of compounds 33a and 33b.

Various methionine metabolites used in the treatment of experimental liver diseases in animals demonstrate hepatoprotective properties. The prophylactic use of choline was shown to provide intensive liver regeneration. In this study, we aim to evaluate the hepatoprotective effects of choline bitartrate in piglets, with an assessment of cytokine expression associated with the hepatobiliary system. The research was conducted using experimental pigs reared at the age of 45 days, which corresponds to the period of their early development. The use of choline bitartrate significantly reduces the expression of β transforming growth factor (TGF-β1) and pro-inflammatory interleukin 6 (IL-6) genes, at the same time as providing a significant reduction in total cholesterol and a proportional increase in HDL concentration, which is a classical anti-atherogenic factor.

This study aimed to evaluate the potential antidepressant effect of ornithine aspartate (LOLA) (3 g/kg/day for 3 months) in mice with an alimentary/toxic model of non-alcoholic steatohepatitis (NASH) (n=70). Animals with NASH demonstrated a significantly decreased sucrose preference, which indicated the presence of food anhedonia. Treatment with LOLA provided a significant anti-anhedonic effect.

An experimental model of acute respiratory distress syndrome was used to determine the following easily reproducible parameters for assessing the efficacy and safety of various glucocorticosteroid therapy regimens: cell viability in bronchoalveolar lavage fluid, calculated values of air-blood barrier permeability index, as well as other routine cytological and biochemical indicators.

The author investigates changes in the indicators of the total electrical activity of the rat brain under conditions of preventive 10-fold UHF exposure during modeling of cerebral ischemia-reperfusion. Preventive UHF exposure was shown to exhibit a neuroprotective effect, manifested in an increase in the total electrical activity of the brain during modeling of ischemia-reperfusion (in particular, at the stages of ischemia after 30 min and reperfusion at the 30th min). This corresponds to the parameters of improving the organization of rhythmic activity of the nervous system.

Xylaria polymorpha mushrooms produce a wide variety of secondary metabolites with unique chemical structures and diverse biological activities. The antimicrobial, antioxidant, and anti-inflammatory properties of Xylaria polymorpha metabolites were previously reported. The present study shows that an in vitro obtained extract obtained from the mycelial biomass of Xylaria polymorpha (Pers.) Grev. exhibits a hypotensive effect, due to an increase in the elastic properties of the main arteries (primarily the root and ascending aorta) and an improvement in endothelial function. This process is accompanied by a decrease in energy costs during cardiac contraction and an increase in the functional reserves of the cardiovascular system.

The aim of this study was to investigate the effect of a Cordyceps militaris (L.) Fr. fungus extract on cardiovascular parameters, including systolic and diastolic blood pressure. The administration of an extract prepared in vitro from a mycelial biomass of Cordyceps militaris (L.) Fr. to male rats was found to increase blood pressure without changing the rigidity of the vascular wall. At the same time, the metabolic and energy processes in the myocardium and the reserve capacity of hemodynamics were increased due to the transition to a more favorable, anabolic variant of metabolism and an economical mode of functioning and expenditure of the body's reserves.

Experiments were performed using 35 male Wistar rats. A metal-organic coordination polymer modified with iron oxide and ascorbic acid (composite) was administered intragastrically to healthy rats at doses of 25 and 50 mg/kg. The intragastric administration of the preparation under study at doses of 25 and 50 mg/kg was found to lead to a significant transient increase in serum iron concentration, while not leading to disruption of pro/antioxidant homeostasis in healthy animals. The intragastric administration of the preparation at a dose of 25 mg/kg, not 50 mg/kg, leads to the accumulation of lymphocytes in the peripheral blood at the late stage of apoptosis.

The imbalance between the production and clearance of β-amyloid peptide (Aβ) from the central nervous system (CNS) is a key factor in pathogenesis of Alzheimer's disease (AD). Human serum albumin (HSA), as a depot of Aβ in the peripheral blood, favors clearance of Aβ from the CNS. HSA carries a variety of ligands that have the potential to affect the HSA-Aβ interaction. Similarly, this interaction can be modulated by Aβ ligands. In our work, we studied the influence of a number of low-molecular-weight ligands of HSA, as well as some regulatory neuroinflammatory proteins with affinity for Aβ, on HSA interaction with monomeric form of Aβ. The established in vitro effects are consistent with clinical data and those obtained in animal models, demonstrating the applicability of our approach in developing new compounds for AD treatment and prevention.