The level of ROS (fluorescent probe 2’,7’-dichlorodihydrofluorescein diacetate) and lipid content (fluorescent lipophilic dye Nile Red) in the peripheral blood monocyte fraction from patients with type 1 diabetes mellitus and healthy volunteers were assessed by flow cytofluorimetry. The number of CD36+ monocytes was assessed using specific antibodies. In patients with type 1 diabetes mellitus, the levels of ROS and intracellular lipids in monocytes and the number of cells expressing CD36 fatty acid translocase were elevated. These results indicate metabolic changes in the peripheral blood cells of patients with carbohydrate metabolism disorders and can be considered as possible prognostic markers for the development of type 1 diabetes mellitus complications.

The development of novel strategies for diagnosing, treating, and preventing cardiovascular diseases (CVDs) linked to metabolic syndrome and obesity presents a significant challenge for the scientific community. There is a pressing need to identify effective compounds that target the underlying pathogenic mechanisms of these disorders. Increasing knowledge about the pathogenesis of CVDs has highlighted the crucial role of perivascular adipose tissue (PVAT) in maintaining cardiovascular homeostasis. PVAT is a metabolically active endocrine organ that plays a key role in regulating blood vessel tone, endothelial function, and the growth and proliferation of vascular smooth muscle cells. However, in metabolic disorders, there is a disruption in the functional activity of PVAT cellular components and an imbalance in the production of vasoactive substances, leading to the development and progression of CVDs. This review systematically examines the morphofunctional changes in PVAT associated with metabolic syndrome and obesity, emphasizes the dysfunction of PVAT as a key pathogenetic factor in cardiovascular disease, and evaluates the potential of hydrogen sulfide (H2S) produced by PVAT as a promising vasoregulatory agent based on existing data.

Aim. To examine the pattern of morphological changes, RNA quality number, and gene expression in mouse tissues sampled at autopsy under controlled experimental conditions.Materials and methods. Balb/c mice were euthanized and subsequently subjected to necropsy at 0, 3, 12, 24, 48, and 72 hours of the postmortem period. During the first three hours following euthanasia, the mice were maintained at room temperature, after which they were transferred to a refrigerator (4 º С). Total RNA was extracted from tissue samples taken from the kidney, liver, and brain; the integrity of the RNA samples was assessed by capillary electrophoresis, and the RNA quality number (RQN) was calculated. The expression levels of Actb, Epas1, and Rps18 housekeeping genes were evaluated by real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) with original primers and probes using the TaqMan assay. The histologic examination was performed according to standard techniques.Results. Degradation of RNA extracted from mouse kidney tissues appeared to be greater than that of RNA taken from the liver. In the meantime, a negative linear correlation was observed between RQN and the duration of the postmortem interval for liver and kidney samples. In contrast, no significant changes in the RQN score were observed for brain RNA samples at any of the time points. The expression of the Epas1 and Rps18 genes was significantly decreased in mouse kidney and liver tissues. However, the level of Epas1 and Rps18 gene expression in the brain remained stable at all time points and did not exhibit a significant decrease at 72 hours after euthanasia. No obvious morphological changes were detected by the histologic examination, which does not exclude the presence of ultrastructural pathological changes.Conclusion. RQN in autopsy tissues serves as a crucial predictor of sample quality for molecular biology studies, including gene expression analysis.

Introduction. Biologically active substances of plant origin are the subject of study in the context of search and development of new pharmacologi-cal agents capable of influencing cholesterol metabolism in the body. The article presents the results of safety assessment, pharmacokinetics and mechanism of pharmacological activity of a new promising hypocholesterolemic agent – L-rhamnopyranosyl-6-O-methyl-galacturonan, a polysaccharide isolated from the leaves of birch (Betula pendula Roth.). Objective of the study – comprehensive study of pharmacokinetic parameters, safety and mechanisms of activity of L-rhamnopyranosyl-6-O-methyl-galacturonan in vivo and in vitro. Material and Methods. Evaluation of acute toxicity by single intragastric or intraperitoneal administration was performed on BALB/c mice and SD rats (Sprague-Dawley). To determine the effect of polysaccharide on bile acid excretion in rats with experimental hyperlipidaemia, faeces were collected for bile acid determination. Blood plasma was used in the evaluation of pharmacokinetics. Detection was performed using high-performance liquid chro-matography mass spectrometry method. To assess the sorption activity of polysaccharide, polysaccharide or a comparison drug cholestyramine was added to a solution of cholic or deoxycholic acid, unbound bile acids were quantified. Light microscopy was used to visualise polysaccharide-bile acid complexes. Results. After intragastric administration of polysaccharide at a dose of 1500 mg/kg the object of the study is practically not subjected to absorption from the digestive tract and can exert its hypolipidemic effect through effects directly in the intestinal lumen. Polysaccharide does not penetrate into organs and tissues and has no systemic action, it is completely excreted through the GI tract. According to the results of acute toxicity experiments the investigated substance can be characterised as practically non-toxic. The mechanism of hypolipidemic action of polysaccharide is associated with its ability to bind bile acids in the intestine, which is confirmed by the obtained data on the increase in the excretion of bile acids with faeces in labora-tory animals receiving polysaccharide, and the established ability of polysaccharide to bind bile acids in vitro. Conclusions. After oral administration, L-rhamnopyranosyl-6-O-methyl-galacturonan is practically not absorbed from the digestive tract, has no toxic effects, and exerts its hypolipidemic effect by binding bile acids in the intestinal lumen

The aim of this review was to analyze the accumulated data on the use of mass spectrometry in diagnosing, treating, and prognosing cancer from the perspective of precision medicine. Currently, universally accepted methods for early cancer diagnosis are not available, primarily due to low molecular specificity of pathological changes at early stages of cancer development. Additionally, the existing diagnostic modalities are notably limited in sensitivity. However, early detection is imperative for selection of the most suitable cancer treatment strategy and its successful implementation. In the realm of oncology, mass spectrometry approaches show great potential for advancement and utilization. Mass spectrometry is becoming an indispensable tool in basic and applied research due to its sensitivity, specificity, and accuracy. It allows for efficient analysis of complex biological compounds, even at low concentrations. Moreover, contemporary mass spectrometry technology is capable of automating the analysis, thereby facilitating its diverse clinical applications in diagnosis, drug therapy selection, and even potential assistance to surgical oncologists in the operating room. Considering all these characteristics and advantages, mass spectrometry methods for the analysis of biological samples can be defined as some of the most promising and dynamically developing tools in precision medicine, as they are capable of providing clinically valuable information based on omics technologies, taking into account personal characteristics of the patient. Over the next decade, introduction of mass spectrometry-based methods into clinical practice based on the principles of precision medicine is expected to optimize selection of personalized treatment strategies for cancer patients and provide significant economic benefits by reducing morbidity, disability, and mortality.This comprehensive review presents the analysis of 65 scientific publications, highlighting the results of clinical and experimental studies utilizing mass spectrometry methods for diagnosing cancer, investigating the underlying mechanisms of disease development, and evaluating the efficacy of therapeutic interventions. The review encompasses original articles published from January 1, 2018 to November 30, 2023. The majority of studies back the potential of mass spectrometry as a valuable tool for cancer diagnosis and treatment monitoring. Broadening application of mass spectrometry techniques in the field of oncology holds significant promise and represents a relevant area for future research.

Aim. To develop a new method to determine the viability of Opisthorchis felineus in vitro using the MTS reagent and to evaluate its applicability for analyzing the efficacy of anthelmintic agents in the treatment of opisthorchiasis.Materials and methods. Golden hamsters were used to create a model of O. felineus infection. The animals were infected with metacercariae obtained from fish of the Cyprinidae family. Three months after infection, adult parasites were extracted from the hepatobiliary system. Their viability was assessed using the motility scale and a new method based on the modified MTS test protocol. To account for differences between the size and number of adult parasite cells, the results were normalized with respect to protein content. To evaluate the feasibility of the new approach in the study of pharmacological activity against opisthorchiasis, the viability of adult parasites in the presence of praziquantel was tested.Results. During incubation of adult flukes in a medium with the addition of the MTS reagent, colored watersoluble formazan was accumulated. Thermal inactivation of parasites significantly decreased the production of this compound. Since the studied adult parasites differed in size and number of cells, the obtained data on their viability were normalized to protein content. The results correlated with the data on parasite viability obtained by the traditional method using the motility scale. Evaluation of praziquantel efficacy at different concentrations using two independent methods (the MTS test and the motility scale) showed that the results of the MTS test were consistent with literature data and comparable with the results obtained using the motility scale.Conclusion. A new method for in vitro evaluation of anti-opisthorchiasis activity of drugs was developed. It is based on the assessment of water-soluble formazan production by adult O. felineus flukes in the culture medium using the MTS reagent for screening anti-opisthorchiasis activity of new anthelmintic drugs.

Phenotypic characteristics of alveolar macrophages in the bronchoalveolar lavage fluid as well as their ability to acquire the M1 and M2 phenotypes during in vitro culturing with reprogramming factors were studied in rats with modeled diet-induced metabolic syndrome. A decrease in the number of alveolar macrophages with the M1 phenotype was found in animals with metabolic syndrome. The factors of metabolic syndrome do not affect phenotypic plasticity of cells in culture, but under the action of M2 reprogramming factors, the cells demonstrate a wide range of phenotypic plasticity by the CD80 and CD206 markers. The consistently high level of production of IL-6 and IL-10 by macrophages during culturing under different conditions indicates functional rigidity of the cells, which is probably a consequence of in vivo predetermined functional phenotype of these cells against the background of metabolic disorders.

Type 1 diabetes mellitus was modeled in Wistar rats by intraperitoneal injection of streptozotocin (25 mg/kg for 5 days), which led to the appearance of the main symptoms of insulin-dependent diabetes. In peripheral blood mononuclear cells isolated by centrifugation on a Ficoll density gradient, the production of ROS and the level of intracellular lipids were evaluated by flow cytofluorimetry. In rats with type 1 diabetes mellitus, an increase in ROS levels in isolated peripheral blood monocytes, but not in the lymphocytic fraction was revealed. Incubation of isolated monocytes in a medium containing 1 mM oleic acid led to a 1.5-fold increase of intracellular lipid levels. After incubation of the lymphocyte fraction in this medium, no differences from the control were revealed. Disorders of carbohydrate and lipid metabolism in type 1 diabetes mellitus leading to an increase of free fatty acids and ROS levels can be detected ex vivo in isolated peripheral blood mononuclear cells.

Obesity is considered as a chronic progressive disease, heterogeneous in its etiology and clinical manifestations, and characterized by excess in body fat mass and its deposition in the body. The term “morbid obesity” refers to excessive deposition of adipose tissue with a body mass index (BMI) ≥40 kg / m 2 or with a BMI ≥ 35 kg / m 2 in the presence of serious complications associated with obesity. Along with obesity, the frequency of type 2 diabetes mellitus and cardiovascular diseases closely associated with it has increased. It results from the progression of metabolic disorders, including insulin resistance, which is inextricably linked with the accumulation of visceral fat and plays a key role in the pathogenesis of obesity-related diseases. The study of lipidomic signatures in obesity and associated conditions is a promising branch of fundamental medicine, which makes it possible to significantly and at a new conceptual level stratify a cohort of obese patients into various phenotypes, including a metabolically healthy and metabolically unhealthy obesity phenotypes. Dynamic changes in the lipidome both in the context of diet, drug treatment, and after various bariatric surgeries are of great interest for developing personalized strategies for the treatment of this disease. Currently available studies and their results suggest that we are only at the very start of studying this promising biomedical field.

The aim of the study was to assess the level of CD68-positive macrophages in adipose tissue in rats with of diet-induced MetS against the background of changes in the concentration of pro-inflammatory cytokines. In animals fed a high-fat and high-carbohydrate diet, there was an increase in body weight, obesity, metabolic disorders, a high concentration of cytokines IL-10 and IL-10, an increase in the content of CD68 macrophages in adipose tissue was observed.

Physicochemical and pharmacological properties of the humic substances from coal of the Kansk-Achinsk coal basin, the Pereyaslovskoe deposit, have been studied. The presence of polycyclic aromatic structures and aliphatic fragments with various functional groups as substituents (carboxyl, carbonyl and quinoid, phenolic, alcoholic, ether and ester, amino and amide) was established by means of UV, IR, 13C NMR spectroscopy, fluor-imetry, and elemental analysis. It has been determined by HPLC that coal humic substances are highly hydro-philic polydisperse biopolymers with medium molecular weights. According to the indicator of microbiological purity (category 3B), the content of microorganisms does not exceed the indicators required by the State Pharmacopeia of the Russian Federation (SPh XIV edition). The content of radionuclides, toxic metals (lead, cadmium, mercury, arsenic) corresponds to the norms (according to SPh XIV edition). The presence of 11 essential elements has been established. The studied humic substances belong to the V class of hazard (low-hazard sub-stances, according to GOST 32644-2014), do not have allergising properties and cytotoxic effect within a broad concentration range. Antioxidant, immunotropic, cytoprotective properties were established in the studies of specific pharmacological activity with various experimental models. The ability to inhibit free radicals such as hydroxyl radicals (HO.) and superoxide-anion radical (O2-. should be stressed, as these radicals are able to by-pass the endogenous antioxidant defense systems of the body. It is concluded that the studied humic substances of the Kansk-Achinsk coal basin, the Pereyaslovskoe deposit, are safe and effective biologically active sub-stances of natural origin for use as a promising biologically active substance in the food and pharmaceutical industries.

In male Syrian hamsters fed a synthetic high-fat diet enriched with cholesterol (0.3%), administration of a polysaccharide from birch leaves L-rhamnopyranosyl-6-O-methyl-D-galacturonan (3 g/100 g of diet) resulted in a decrease in total cholesterol levels, mainly due to the LDL fraction, triglycerides, and bile acids in blood serum; the content of triglycerides and cholesterol in the liver also decreased, while excretion of bile acids with feces increased. Thus, the lipid-lowering effect of L-rhamnopyranosyl-6-O-methyl-D-galacturonan is related to its ability to bind bile acids in the intestine and interrupt their enterohepatic circulation.

We studied the effect of an indolinone derivative GRS on the development of experimental atherosclerosis in C57BL/6 mice. Atherosclerosis was modeled by intraperitoneal administration of endothelial lipoprotein lipase inhibitor Kolliphor P 407 micro Geismar over 5 months. GRS was administered orally in a dose of 10 mg/kg once a day throughout the experiment. In 5 months, the levels of total cholesterol, LDL, and triglycerides in blood serum, as well as histological composition of the ascending aorta were studied. In mice with experimental atherosclerosis, we observed pronounced dyslipidemia with an increase in serum cholesterol, LDL, and triglycerides and accumulation of xanthoma cells in the aorta wall. Repeat administration of GRS did not eliminate dyslipidemia, but prevented an increase in the number of xanthoma cells in the aorta wall (p<0.05). The stimulator of soluble guanylate cyclase GRS did not exhibit hypolipidemic activity, but restored impaired endothelial function in the atherosclerosis model and prevented atherosclerotic damage to blood vessels and vascular wall remodeling.

Aim. To analyze modern methods for the diagnosis of trematodiasis in experimental and epidemiological studies. Trematodiasis is a group of common parasitic diseases that are a socially sensitive health problem worldwide. According to World Health Organization, more than 250 million people are affected by trematode infections globally. The most common types of human trematode infections are diseases caused by Schistosoma, Fasciola, Clonorchis, and Opisthorchis pathogens. Diagnosis of trematodiasis is often multistage and includes identification of disease symptoms, analysis of medical history, and use of various laboratory tests. Clinical presentation of parasitic infections often varies, making a definitive diagnosis difficult. Various tools are used to diagnose trematode infections: epidemiological criteria, laboratory tests (complete blood count and blood biochemistry, serological methods), instrumental methods (abdominal X-ray and ultrasound), and parasitological techniques, which often have insufficient sensitivity and specificity. Therefore, development of modern and effective non-invasive methods for detection of trematode infections with high sensitivity and specificity, including screening in endemic regions, is relevant. The present review analyzes the results of 90 clinical trials and experimental studies on the diagnosis of trematode infections using the PubMed search engine and the eLibrary database. The review analyzes original articles published from January 1, 2015 to December 31, 2021. Most studies confirm that the absence of a standard diagnostic approach highlights obvious convenience of utilizing a combined approach to reliable diagnosis of trematodiasis. An adequate combination of different diagnostic tests makes it possible to diagnose the disease correctly, devise a correct treatment and follow-up strategy, and organize preventive measures.

The aim was to investigate wound-healing properties of zinc-containing biocomposites based on humic ligands (humic substance (HS) – Zn) in the in vivo experiment on the aseptic wound model and to evaluate their resorptive properties. Materials and methods. The objects of the study were 5 samples of HS-Zn in the form of complex salts comprising fine black powders synthesized in the Laboratory for Natural Humic Systems of the Faculty of Chemistry at Moscow State University. The wound-healing effect of the substances was studied on 70 male Wistar rats using a traumatic model of an excisional aseptic skin wound. The degree of affected skin healing was evaluated during 21 days by the planimetric method. The resorptive properties of the HS-Zn samples were studied by inductively coupled plasma mass spectrometry (ICP-MS) in the biomaterial (blood serum, fur, skin from the wound surface). Results. It was found that course application of zinc-containing HS-Zn biocomposites to the wound surface led to a decrease in the wound area in comparison with ZnSO 4 with the equivalent concentration of elemental Zn (1.67 mg/ ml). Two samples FA-Zn and Peat1-Zn showed the most pronounced regenerating effect. We noted an increase in Zn level in the tested skin samples from the wound area, in fur, and in the blood serum, which indicates the resorptive effect of zinc-containing HS-Zn biocomposites during course application; however, the parameters did not exceed limiting permissible concentrations. The correlation between the tested samples was not equal, which indicates a significant impact of the initial HS matrix on the Zn bioavailability. Conclusion. The observed reparative effect of zinc and HS complexes in the context of their low toxicity is of interest for further study to develop effective wound-healing preparations.

Aim: To study the functional state of the components of the glutathione-dependent antioxidant system in the adipose tissue of rats with experimental metabolic syndrome (MetS).Material and Methods. The MetS model was carried out on male Wistar rats using a high-fat, high-carbohydrate diet (HFHCD). Body and adipose tissue weight were measured. Blood serum levels of glucose, insulin, leptin, triacylglycerides and cholesterol were assessed. In epididymal adipose tissue the level of reactive oxygen species (ROS) was determined by fluorescent method. The concentration of reduced (GSH) and oxidized (GSSG) glutathione, activity of glutathione reductase, glutathione peroxidase, and glutathione-S-transferase enzymes were assessed spectrophotometrically in epididymal adipose tissue.Results. It was found that HFHCD led to an increase in body weight, obesity, hyperglycemia, insulin resistance, dyslipidemia, and leptinemia in the experimental group rats. An increase in adipose tissue mass had a positive correlation with an increase in the concentration of glucose, serum leptin, and ROS levels in the epididymal adipose tissue of rats with MetS. It was found that the level of total glutathione in the adipose tissue of the experimental group rats decreased mainly due to a decrease in the level of GSH. The rats receiving HFHCD also showed a decrease in the activity of glutathione peroxidase and glutathione-S-transferase, but the activity of glutathione reductase increased.Conclusion. Obesity, as a key component of MetS, is a trigger of insulin resistance, chronic low-grade inflammation and oxidative stress. The study showed that the development of MetS and obesity in the experimental animal group is accompanied by a shift of adipocyte redox balance toward oxidative stress, which is expressed in a decrease of GSH/GSSG ratio and glutathione-dependent antiperoxide protection enzymes activity.

Background. Overweight and obesity are key factors for the occurrence of many morphofunctional disorders in organs and tissues, including bronchopulmonary system.The aim. To study the influence of metabolic disorders that occur against the background of obesity on the state of the airways tone in rats.Materials and methods. Obesity in male Wistar rats was induced using a high-fat and high-carbohydrate diet. In animals, body weight and fat mass were measured, and the heart-lung complex was extracted. In blood serum, the levels of glucose, insulin, leptin, triglycerides, and cholesterol were assessed. Bronchoalveolar lavage fluid was obtained by an open method, in which the concentration of protein, interleukin (IL) 6 and IL-10 was determined. The contractile activity of the isolated bronchial smooth muscle segments was studied using mechanographic method. The effect of acetylcholine (10–7–10–4 M), indomethacin (10–5 M), and forskolin (10–7–10–5 M) on the changes in the tone of airway smooth muscles was assessed.Results. High-fat and high-carbohydrate diet caused an increase in body weight, visceral obesity, hyperglycemia, insulin resistance, leptinemia, dyslipidemia in rats of the experimental group. In the bronchoalveolar lavage fluid of experimental animals, an increase in the content of protein and IL-6 was found, which positively correlated with the level of leptin and the fat mass. In obese rats, the contractile responses of bronchial smooth muscle segments increased in response to the effect of the cholinergic agent acetylcholine. The bronchoconstrictor effect of acetylcholine was reduced by the cyclooxygenase inhibitor indomethacin. In turn, the adenylate cyclase activator forskolin caused relaxation of the airway segments smooth muscles in rats of both groups, which was more pronounced in the experimental group.Conclusion. The obtained results indicate that the change in the reactivity of the respiratory tract can be the cause of bronchospastic conditions in obesity and of the inflammatory reaction in the respiratory system induced by obesity.

We studied contractile responses of isolated airway smooth muscle segments from rats with metabolic syndrome. Metabolic syndrome was induced in rats by high-fat and high-carbohydrate diet. It was shown that metabolic syndrome was associated with an increase of bronchoconstrictor action of cholinergic receptor activator carbacholine (0.1-100 μM) and a decrease of the dilatory effect of β2-adrenoreceptor activator salbutamol (0.1-100 μM). The observed effects of agonists are epithelium-dependent. Disorders in contractile activity in the airway smooth muscles were accompanied by bronchial epithelium destruction, immune inflammation in the bronchial wall, muscular and peribronchial adipose tissue hypertrophy.

Introduction . Dyslipidemia treatment in many cases requires carefully selected combination of lipid-lowering drugs including bile acid sequestrants. A promising compound is L-rhamnopyranosyl-6- O -methyl-galacturonan (L-RAG), a polysaccharide obtained from the birch leaves ( Betula pendula L.). Aim . To evaluate the lipid-lowering activity of L-RAG administered in combination with the HMG-CoA reductase inhibitor rosuvastatin or the cholesterol absorption inhibitor ezetimibe. Materials and methods . Hyperlipidemia in hamsters was reproduced by synthetic diet supplemented with 0.3 % cholesterol and 11 % coconut oil (HFCD). At the end of the experiment the levels of triacylglycerols (TAG), bile acids, total cholesterol (TC), cholesterol of low-density lipoproteins (LDL-C) and high-density lipoproteins (HDL-C) were determined in the blood serum using Chronolab Systems S. L. enzymatic kits (Spain), and atherogenic index (AI) was calculated. Results and discussion . L-RAG demonstrates a lipid-lowering effect in a model of diet-induced hyperlipidemia in hamsters. The decrease in the TC level was mainly caused by a decrease in the content of cholesterol in the atherogenic LDL fraction, which led to a decrease in AI. The use of rosuvastatin and ezetimibe in combination with L-RAG leads to a better lipid-lowering effect. The additive effect of the polysaccharide is mainly due to a decrease in the level of bile acids in the blood serum, which indicates its ability to absorb them in the intestinal lumen and interrupt enterohepatic circulation, acting like bile acid sequestrants. Conclusion . It was shown that administration of L-RAG together with ezetimibe or rosuvastatin led to a better decrease in the TC, TAG, LDL-C levels than with polysaccharide monotherapy or comparison drugs alone. The obtained data substantiate the prospects of using L-RAG in the complex therapy of hyperlipidemia.