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Vol 25, No 5 (2022)

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Biological chemistry

Glucogenic and ketogenic effects in the light of introducing their skeletons into Krebs cycle

Malinovsky A.V.


After catabolism of all amino acids became known, attempts to make up a scheme of ways of introducing amino acids'carbon skeletons in different stages of oxydative transformations in Krebs cycle were made several times; amino acids having been united in groups. However, all schemes which are known so far have some faults particularly referring to glucogenic or ketogenic effects of one or another amino acid. Moreover, the conclusion to which the scientists came with reference to this effect, do not always correspond to the scheme. Faults both in the sceme and conclusions concerning glucogenic and ketogenic effects of amino acids are not admissible; these effects can be clearly observed in case of diabetes: introduction of glucogenic amino acids into the organism reduces the severity of ketosis whereras that of ketogenic increases it. In the present paper a scheme of introducing amino acids'carbon skeletons into Krebs cycle, specified on the basis of modern data of amino acids' transformation, and conclusions of glucogenic or ketogenic effects of one or another amino acid that fully correspond to the scheme are given. The results should be taken into account when dealing with diabetes and some other diseases.

Problems of Biological Medical and Pharmaceutical Chemistry. 2022;25(5):3-11
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Production of interleukin-6 by resting hybridoma endothelial cells of the EA.HY926 line

Saginbaev U.R.


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.

Problems of Biological Medical and Pharmaceutical Chemistry. 2022;25(5):12-15
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Medical chemistry

The concentration of glucose, zinc cations, and indicators of oxidative status in saliva as non-invasive markers of type 2 diabetes mellitus

Cheprasova A.A., Popov S.S., Pashkov A.N., Kryl’skii E.D., Verevkin A.N.


Relevance. One of the acute problems of biomedicine at present is type 2 diabetes mellitus (DM2), the prevalence of which is constantly growing in the Russian Federation and other countries. One of the key pathogenetic factors of this pathology is oxidative stress, which plays a central role in the development of diabetes complications. The need for regular monitoring of the course of DM2 makes it urgent to search for new objects for non-invasive assessment of glycemia and additional parameters that can reflect the severity of the pathological process. A number of studies on the analysis of the relationship between the biochemical composition of saliva and blood plasma determine the expediency of using saliva as an object for assessing the level of glycemia and the state of redox homeostasis. In the course of this work, we studied the correlations between indicators of the oxidative status, the content of zinc ions in saliva, and the level of glucose in the blood serum of patients with DM2. Material and methods. The study included 80 people, the experimental group consisted of DM2 patients divided into 2 subgroups: 20 men (age 62.60±1.92 years) and 20 women (62.85±2.78). The control group included 2 subgroups: 20 apparently healthy men (41.05±3.54) and 20 women (43.35±3.18). Mixed saliva was collected in the morning on an empty stomach. The concentration of glucose, diene conjugates, zinc cations, superoxide dismutase and catalase activity were determined in saliva. Analysis of the oxidative status was also carried out on the basis of biochemiluminescence parameters. Results. The results of the study showed that the saliva of patients with DM2 is characterized by reduced activity of antioxidant enzymes and the concentration of zinc cations, as well as increased levels of oxidative stress. Positive correlations were established between the parameters of blood serum glycemia and indicators of the redox status of saliva, both in healthy participants and in sick donors. Conclusion. The data obtained indicate that saliva can be used as a biomaterial for diagnosing T2DM and monitoring the effectiveness of treatment for this disease.

Problems of Biological Medical and Pharmaceutical Chemistry. 2022;25(5):16-21
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Assessment of the selectins and their ligand PSGL-1 level in patients with acute venous thrombosis

Korotkova N.V., Kalinin R.E., Suchkov I.A., Mzhavanadze N.D., Nikiforov A.A., Romanov B.K., Bodrova O.V.


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.

Problems of Biological Medical and Pharmaceutical Chemistry. 2022;25(5):22-27
pages 22-27 views

Pharmaceutical chemistry

Standardization of anti-inflammatory herb tea

Lubsundorzhieva P.B., Kashenko N.I., Dargaeva T.D.


The composition of anti-inflammatory herbal tea intended for the treatment and prevention of chronic colitis includes Bergenia crassifolia (L.) Fritsch. black leaves, Matricaria chamomilla L. flowers, Achillea millefolium L. herb and Mentha piperita L. leaves. Standardization of anti-inflammatory herbal tea is the purpose of this work. Material and methods. Samples of herbal tea produced by SMEs "Arura" (Ulan-Ude) were used. The microcolumn HPLC method was used to investigate the composition of phenolics. The total contents of biologically active substances were determined according to pharmacopoeial methods. Results. The contents of arbutin, gallic acid, bergenin, chlorogenic acid, rutin, rosmarinic acid in herbal tea were 1.10, 0.12, 0.34, 0.05, 0.46 %, respectively. The total contents of flavonoids, phenolic acids, tannins, triterpenes, anthocyanins, organic acids, water-soluble polysaccharides, pectin substances, amino acids in herbal tea were 2.96, 0.21, 9.45, 1.24, 0.21, 3.90, 4.10, 0.90, 0.14%, respectively. A differential method for the quantitative determination of flavonoids was adapted to standardize the herbal tea, the error of a single determination of the content of flavonoids did not exceed ±4.45%. Diagnostically significant microscopic features of herbal tea were established. Minimal content levels of the main groups of biologically active substances in anti-inflammatory herbal tea have been established. Thus, total contents of flavonoids (luteolin equivalents), tannins and arbutin were not less than 2.0, 8.0, 0.5%, respectively. Conclusions. Anti-inflammatory herbal tea is proposed to standardize according to the sum of phenolics: flavonoids (luteolin equivalents), tannins and arbutin. Minimal content levels of the main phenolics, diagnostically significant microscopic features of herbal tea were established.

Problems of Biological Medical and Pharmaceutical Chemistry. 2022;25(5):28-34
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The use of HPLC-MS/MS for the quantitative analysis of neuroactive amino acids in rat brain homogenates after derivatization with 9-fluorenylmethyl chloroformate

Popov N.S., Balabanyan V.Y., Petrova M.B., Kolgina N.Y., Petrov G.A.


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.

Problems of Biological Medical and Pharmaceutical Chemistry. 2022;25(5):35-45
pages 35-45 views

Study of the influence of structural and mechanical parameters of gelling agents for the creation of soft dosage forms with ginkgo biloba extract

Kovtun E.V.


Relevance. The issues of creating domestic phytopreparations closely related to the problem of import substitution in pharmacy are significant. In this regard, the development of the composition and technology of nasal gel, using dry ginkgo biloba extract as an active component, for the treatment of chronic disorders of cerebral circulation, is very relevant. The purpose of study is to analyze the structural and mechanical parameters of the dosage form, the effect of excipients and active substance, to create a dosage form with the ginkgo biloba phytocomplex. Material and methods. To assess the effect of different concentrations of the chitosan gel-forming agent and the active substance on the structural and mechanical parameters of the dosage form, model compositions of gels were studied. Rheological characteristics of the samples were studied by rotational viscometry. Conclusion. Rheological characteristics of the dosage form have been established. The effect of the concentration of chitosan gel-forming agent and the active substance of dry ginkgo biloba extract on the structural and mechanical parameters of the dosage form was studied. The viscosity of solutions of high-molecular compounds was determined.

Problems of Biological Medical and Pharmaceutical Chemistry. 2022;25(5):46-51
pages 46-51 views

In vivo comparison of the biological activity of topical methylprednisolone aceponate formulations using vasoconstriction assay

Nesterov M.S., Khvostov D.V., Ageldinov R.A.


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.

Problems of Biological Medical and Pharmaceutical Chemistry. 2022;25(5):52-58
pages 52-58 views

Anniversaries and dates

Scientific heritage of academician of the RAS Yuri Ivanovich Denisov-Nikolsky (on the 90th anniversary)

Matveichyuk I.V., Krasnov V.V., Rozanov V.V.


The article presents information about the main stages of scientific, practical, pedagogical and social activities of the outstanding Soviet and Russian morphologist, Honored Scientist of the Russian Federation, Academician of the Russian Academy of Sciences, Doctor of Medical Sciences, Professor Yu.I. Denisov-Nikolsky (1932-2018). His significant contribution to domestic and world science, to the achievements of the FGBNU VILAR, to the development of many theoretical and applied problems of modern morphology of the of the osteoarticular apparatus is shown. The importance of Yu.I. Denisov-Nikolsky for the successful development of a new direction he created - "Reproduction of cells, tissues and bioprosthetics", which he successfully led and coordinated research in this area for many years is pointed out.

Problems of Biological Medical and Pharmaceutical Chemistry. 2022;25(5):59-62
pages 59-62 views

Anatoliy Viktorovich Skal'nyy (k 60-letiyu so dnya rozhdeniya)


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.

Problems of Biological Medical and Pharmaceutical Chemistry. 2022;25(5):63-64
pages 63-64 views

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