Vol 25, No 5 (2022)

Relevance. To study the mechanisms of atherogenesis, experimental models of cell culture (from the umbilical vein (HUVEC) and EA.Hy926 line cells, which are a hybrid of endotheliocytes from the umbilical vein and lung carcinoma), are widely used. The degree of effect of both individual and group cytokines on endotheliocyte physiology can be determined by adding adequate concentrations of mediators to the cell culture. However, to obtain correct results, it is necessary to determine the initial background cytokine concentration characteristic of this cell culture. The purpose of this work was to study the concentration of interleukin 6 in the culture of EA.Hy926 cells. Material and methods. The material was cell cultures of the EA.Hy926 line, which are a hybridoma of endotheliocytes from the umbilical vein (HUVEC) and lung carcinoma (A549). Results. A concentration of IL-6 was found that exceeded by more than an order of magnitude the level of this cytokine in the blood of a healthy person. Conclusion. Thus, when assessing the level of proinflammatory cytokine in the cell model, significant excesses were noted, which requires taking this factor into account during experimental work.

Relevance. Currently, the violation of the adhesive function of the endothelium is considered one of the components of endothelial dysfunction, and therefore is in the center of attention of doctors of various specialties. Violation of the adhesive function is manifested by a change in the synthesis of cell adhesion molecules: hyper- or hypo-expression of the latter leads to a change in the adhesion cascade of leukocytes and disruption of their interactions with endotheliocytes. Aim. To evaluate the concentration of cell adhesion molecules - selectins P, E, L and their glycoprotein ligand in the blood serum of patients with acute venous thrombosis. Materials and Methods. The study included 20 patients whith acute venous thrombosis and 12 subjects were used as control (healthy volunteers). The participants of the study were comparable in age and gender. Serum samples were isolated from peripheral venous whole blood samples. Quantitative measurement of P-selectin and its glycoprotein ligand PSGL-1 was performed using the Sandwich Enzyme-Linked ImmunoSorbent Assay (ELISA) with ELISA Kit for Selectin, Platelet (SELP) (catalogue № SEA569Hu), PSGL-1 ELISA kit (catalogue № SEA985Hu), and Stat Fax 2100 analyzer (microplate reader) (Awareness technology Inc. PalmCity, FL 34990, USA). Results. The concentration of selectin P in peripheral blood in patients with acute venous thrombosis statistically significant decrease by 2.5 times was discovered. The concentration of selectin E showed a downward trend, but it was not statistically significant, which may indicate a lesser degree of involvement of Е selectin in the pathogenesis of venous thrombosis. The concentration of selectin L tended to increase. Concentrations of the glycoprotein ligand of selectins PSGL-1 were statistically significantly increased in the peripheral blood of patients with acute venous thrombosis. Conclusions. Acute venous thrombosis is accompanied by a statistically significant decrease in the concentration of P-selectin, a tendency to decrease in E-selectin, an increase in the concentrations of L selectin and the ligand of selectins PSGL-1 in the blood serum.

Relevance. The effect on the metabolism of neuroactive amino acids is the most important component of the mechanism of action of psychotropic drugs. Variation of substance of amino acids in the structures of the rat brain can act as a pharmacodynamic marker diagnostic sign in the study of the pathogenesis of diseases of the central nervous system. Purpose of the study. development of HPLC-MS / MS method for quantitative determination of neuroactive amino acids in rat brain homogenates after derivatization with 9-fluorenylmethylchloroformate. Material and methods. For the detection of amino acids from the rat brain was used a Potter-Elvehjem homogenizer. Deproteinization and derivatization were performed by adding a solution of 9-fluorenylmethylchloroformate in acetonitrile to the samples. Amino acid derivatives were detected using a Sciex 3200 mass spectrometer. For chromatographic separation was used an Agilent 1260 Infinity II HPLC. Elution was carried out with a mixture of acetonitrile and water in a gradient mode. Results. Sample preparation includes mixing 100 gl of rat brain tissue homogenate, 100 gl of borate buffer, 20 gl of 1 mM norvaline solution and 250 gl of 12 mM Fmoc-Cl solution in acetonitrile, followed by centrifugation for 10 minutes. For the separate Fmoc-derived amino acids was used hromatographic column InfinityLab Poroshell 120 EC-C18 4.6 x 100 mm, 2.7 gm. The total time of chromatographic analysis was 10 minutes, the retention time of Fmoc derivatives of glycine, GABA, aspartic and glutamic acids, asparagine and glutamine was 6.7; 6.8; 6.4; 6.4; 6.2 and 6.1 minutes, respectively. The analytical range of the method for each amino acid was from 0.05 to 50 nmol in 1 ml of homogenate. The method was tested by analyzing the amino acid content in the brain of 6 intact Wister rats. Conclusion. A chromatography-mass spectrometric method for the quantitative determination of glutamine, asparagine, glycine, GABA, glutamic and aspartic acids in rat brain homogenates has been developed. Precolumn derivatization of amino acids with 9-fluorenylmethylchloroformate was carried out to increase the sensitivity of the analysis.

Methylprednisolone aceponate (MPA) is potent (class III) topical corticosteroid widely used for the treatment of inflammatory dermatoses. It's peculiarities are exceptionally favorable safety profile, high anti-inflammatory activity and properties of prodrug - after application to the skin MPA is transformed into active metabolite methylprednisolone 17-propionate, and this process goes more intensively in the zone of inflammation thus limiting steroid effect on intact skin. After the patent expiration generic MPA drugs which didn't pass through the full cycle of clinical trials have entered some markets, in cluding Russian. Metabolic profile of these generics and it's equivalence to the original product is unknown. Objective. To assess biological activity of various drug formulations of original (Advantan®) and generic (Komfoderm®) drugs of MPA using vasoconstriction assay in the in vivo model. Methods. Recommended doses of studied formulations were applied to the skin of laboratory animals (mini-pigs), and then experimental measurements of chromometric data were performed at specific timepoints (0.5 h, 3h, 6 h, 12 h, 24 h) using chromometer FRU WR-10 (8 mm). Results. Difference in the profile of biologic activity based on vasoconstriction assay data was noticed for all tested MPA drugs, ranging in descending order for lightness parameter L: Advantan® cream - Advantan® milk - Advantan® fatty ointment - Advantan® ointment - Komfoderm® K cream - Komfoderm® ointment - Komfoderm ® M2 ointment. Conclusions. Applicability of colorimetric method for evaluation of intensity of biologic effect of glucocorticoids based on the level of lightness of skin surface confirmed. Differences for this parameter for studied drugs detected. Noticed tendency to higher activity of the original drug in comparison to the generic.

On May 12, 2022, Professor Anatoly Viktorovich Skalny, a world-known scientist, a specialist in the study of chemical elements in medicine and biology, the founder of the scientific school of medical elementology, celebrated his 60th birthday.