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Vol 22, No 4 (2019)


The comparative phytochemical and microbiological study of liquid extracts based on the fruits of the Crataegus sanguinea Pall. and Crataegus submollis Sarg

Kurkin V.A., Morozova T.V., Shaikhutdinov I.K., Lyamin A.V., Pravdivtseva O.E., Pervushkin S.V., Kretova A.A.


The preparations based on hawthorn fruits have cardiotonic action. One of the reasons of chronic cardiovascular pathology are various inflammatory processes in the human body. In this regard it is actual the study the antimicrobial activity of preparations based on the raw materials of various species of the Hawthorn genus (Crataegus L.) among which hawthorn blood-red and Quebec hawthorn are promising. Hawthorn blood-red (Crataegus sanguinea Pall.) is a wild-growing plant which is wide-spread in the territory of our country. Quebec hawthorn (Crataegus submollis Sarg.) is widely cultivated on the territory of the Russian Federation and it is characterized by rapid growth and high yield. Liquid extracts were obtained based on the fruits of two species of hawthorn in a ratio of 1:1 using 70% ethyl alcohol. The determination of the minimal inhibiting concentration was carried out by double serial dilution in broth. As test cultures, we used the following microorganisms: Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli, Bacillus cereus, Candida albicans. The study of antimicrobial activity against several microorganisms made it possible to reveal the moderate antimicrobial activity of liquid extracts based on the fruits of hawthorn blood-red and Quebec hawthorn. In addition to the above the antifungal activity of the extract based on hawthorn blood-red fruits is more expressed.
Problems of Biological Medical and Pharmaceutical Chemistry. 2019;22(4):3-6
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Development of the method of quantitative analysis of penolic compounds in herb moldavian draconhead (Dracocephalum moldavica L.)

Zvezdina E.V., Sheichenko O.P.


The purpose of the work is selection of optimal conditions for methods of authenticating and quantifying the amount of phenolic compounds in the herb of the moldavian draconhead (Dracocephalum moldavica L.) Material and methods. The objects of the study were samples of the dried above-ground part of the Dracocephalum moldavica L., harvested in the Botanical Garden, the Middle Volga and North Caucasus branches of All-Russian Scientific Research Institute of Medicinal and Aromatic Plants from 2016-2018. To characterize authenticity of this raw material, a sensitive reaction with 3% solution of iron chloride was used for determine action of phenolic compounds. To determine rosmarinic acid, thin layer chromatography on sorbent (TLC) was used. Sorbfil PTSH-PA plates measuring 10 x 15 were used as the stationary phase. Chromatography was carried out in an ascending fashion using an ethyl acetate - acetic acid - water solvent system (40: 5: 5). In experiments were used standard sample (CO) of rosmarinic acid (Sigma Aldrich, cat. No. 536954-5G). UV absorption spectra were recorded on spectrophotometers UV-1800 (Shimadsu), Cary 100 Scan (Varian). Results and conclusions. Dracocephalum moldavica L. is prospective source for production of new medicinal remedies. Results of the methods of their products are worked out in order to worked out of their identity using novel quality tests and TLC. There was found condition and there was worked out methods of qualitative determination of the total phenolic compounds calculated on rosmarinic acids in Dracocephalum moldavica L. herb using direct spectrophotometric method and their validation was carried out according to the following indicators: specificity, linearity, correctness, intralaboratory precision: convergence and reproducibility.
Problems of Biological Medical and Pharmaceutical Chemistry. 2019;22(4):7-12
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Modern method of elemental analysis of mineralized tissue using energy dispersive x-ray spectroscopy

Litvinov Y.Y.


The results of experimentally tested modern method of elemental analysis of mineralized tissue using energy dispersive x-ray spectroscopy are presented. Material and methods: it have been studied 4 groups of bone samples of 5 pieces each: 1) native bone samples (control group); 2) bone samples, after incubation in hydrochloric acid 0.8 n for 24 hours; 3) bone samples, after incubation in hydrochloric acid 0.8 n for 48 hours; 4) bone samples, after incubation in hydrochloric acid 0.8 n for 72 hours. Evaluation and control of the degree of demineralization of bone implants was performed using scanning electron microscope Phenom XL (Phenom-World, the Netherlands), equipped with a detector for integrated energy dispersive x-ray spectroscopy. Results and discussion: the control showed that after 72 hours of incubation in hydrochloric acid 0.8 n, the presence of - CA, Na, Mg is not determined in the bone matrix according to x-ray spectroscopy. The relative percentage of P has decreased by 45 times. The relative percentage of N has increased by 2.5 times and C has increased by 4.5 times. Summary: experimentally tested method of elemental analysis using energy dispersive x-ray spectroscopy can be used to control the degree of demineralization of bone tissue in the preparation of bone implants for bioimplantology.
Problems of Biological Medical and Pharmaceutical Chemistry. 2019;22(4):13-23
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GC-MS determination of non-volatile components of diluents in plastic smoking blends containing synthetic cannabinoids

Oberenko A.V., Kachin S.V., Sagalakov S.A.


Over the last decade a spread of novel psychoactive substances including synthetic cannabinoids (SC) has become a serious social problem. SC cause addiction and present a danger to life and health of people. Cutting agents are substances used to dilute drugs, psychotropic and potent substances in illicit trafficking. Determination of the qualitative and quantitative composition of the cutting agents has a high forensic significance as for assessing the potential risk to the health of consumers, as for establishing the generality of the source of manufacture of samples containing narcotic drugs. The aim of this study is the GC-MS determination of non-volatile organic components of cutting agents in SC-containing plastic smoking mixtures with the preparation of appropriate derivatives of non-volatile organic components. SC-containing plastic smoking mixtures, had withdrawn from the illegal circulation of the Transport Administration of the Ministry of Internal Affairs of Russia in the Siberian Federal District in the period from 2016 to 2018, were used. 15 samples were analyzed. Sample preparation. The dry residue of water-soluble substances was treated with derivatizing reagents: 0.2 ml of pyridine solution containing hydroxylamine hydrochloride at a concentration of 25 mg/ml was added to the dry residue. Then solution was kept at a temperature of 70 °C until the precipitate dissolved. Then, 10 ^l of BSTFA (NO-bis (trimethylsilyl) trifluoroacetamide) was added to the solution and the mixture was kept at 70 °C for 30 minutes. GC-MS-analysis. Gas chromatography-mass analysis was made on chromatograph "Kristall 5000.2" (Russia) with a quadrupole mass-spectrometer detector "ISQ". Peak identification was based on a comparison of their mass spectra with the data of the mass-spectral library NIST14 and retention times of the standard samples. Results. The proposed procedure of sample preparation and GC-MS method may be a suitable tool for the qualitative analysis of the non-volatile organic components found in cutting agents of SC-containing plastic smoking mixtures. Polyols, monosaccharides, disaccharides and hydroxy acids were found in most of the studied samples.
Problems of Biological Medical and Pharmaceutical Chemistry. 2019;22(4):24-28
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Chromatographic purification of bovine testicular hyaluronidase

Melnikova I.V., Yashtubaeva A.D., Glazova N.V.


Hyaluronidase is an enzyme that breaks down the acidic mucopolysaccharides which are the basis of connective tissue. Integration and analysis of the literature data of different authors, that presented in the article [1], and comparative analysis of the properties of hyaluronidase from different sources, that was carried out at the Department of Biotechnology SPCPU [2], showed that hyaluroni-dases isolated from various sources possess different physicochemical characteristics such as molecular weight, the range of enzymatic activities, stability etc. Therefore, the choice of raw materials determines the approaches to hyaluronidase isolation and purification. In this investigation the bovine testes were chosen as a raw material, since currently only this raw material is used to produce hyaluroni-dase on an industrial scale. Studies of ion exchange sorption of hyaluronidase from the bovine testes showed that the enzyme is selectively sorbed on macroporous sulfocationite KU-23 [3, 4]. The use of this chromatographic method made it possible to replace the unsustainable conventional method of isolating hyaluronidase using organic solvents. However, ionite KU-23 is currently commercially unavailable, that's why the selection of modern sorbent for the isolation and purification of hyaluronidase is a crucial task. The aim of this investigation was to develop a chromatographic procedure of hyaluronidase purification from test solution with the use of modern macroporous sorbents. Materials and methods. The object of research - hyaluronidase in active pharmaceutical ingredient «Lidaza», which is manufactured by «Samson-Med". The object of research is a test solution of hyaluronidase with a protein concentration 10 mg / ml and pH 4.0-4.5. Methods: method for determination of protein with the use of biuret reagent [5]; method for determination of hyaluronidase activity in solution [6]; method for preparation of sorbents [7, 8]; method of chromatographic procedure under static and dynamic condition. Conclusion. The study of chromatographic purification of hyaluronidase under static and dynamic conditions with the use of macroporous sorbents revealed that the optimal sorbent is Nuvia™ S (Bio-Rad Laboratories, Inc). That sorbent provides for the maximum of the protein and hyaluronidase activity yield. Hereafter Nuvia™ S can be used for isolation and purification of hyaluronidase from the orchic extract. New technology can replace conventional technology that uses organic solvents.
Problems of Biological Medical and Pharmaceutical Chemistry. 2019;22(4):29-34
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The frequency of somatic mutations in EGFR gene in smoking and non-smoking patients with lung adenocarcinoma

Badaev E.A., Fazlov V.V., Kvon T.A., Gulyaeva L.F.


Lung cancer is one of the most common malignancies in the world. On the territory of the Russian Federation, in particular the Novosibirsk region, there is an increase in the number of morbidity and mortality from lung cancer. There are many factors leading to the development of this disease, such as smoking, environment, etc. One of the possible options for the development of lung cancer are activating mutations in the gene for the receptor of epidermal growth factor (EGFR). Activation of EGFR plays a crucial role in tumor proliferation, survival and metastasis, so EGFR is a target for targeted therapy. The frequency of activating mutations in the EGFR gene depends on many factors, including regional features. For example, the frequency of such mutations is particularly high in nonSmoking women in East Asia. The purpose of this study is the analysis of the prevalence of non-small cell lung cancer, particularly adenocarcinoma of the lung in the Novosibirsk region, and the frequency of its occurrence depending on the Smoking and the presence of mutations in EGFR in men (n=352) women (n=197). The results of the study showed that lung adenocarcinoma occurs in male smokers in 52.3%, and non-smokers in 43.5%. At the same time, mutation in EGFR in non-Smoking men (6%) was higher than in smokers (2.6%). In the study of the female group, the incidence of lung adenocarcinoma in Smoking women was 6.2%, and in non -smokers- 92.6%. Mutation in EGFR was detected in 35.5% of non-smoking patients, while smoking women with lung adenocarcinoma, mutation in EGFR was not detected. Thus, the results indicate different pathogenetic mechanisms of lung cancer development in men and women, depending on Smoking and the presence of mutations in the EGFR gene. A feature is a fairly high frequency of mutations of EGFR (up to 35.5%) in non-Smoking women with lung adenocarcinoma in the Novosibirsk region.
Problems of Biological Medical and Pharmaceutical Chemistry. 2019;22(4):35-42
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Contents of volatile fatty acids in condensate of exhaused air in smoking persons

Karavaeva T.M., Tereshkov P.P., Maximienya M.V.


Smoking annually leads to nearly six million deaths. Exposure of tobacco smoke components to cells leads to significant metabolic shifts, including lipid metabolism disorders. Volatile fatty acids are important biologically active substances, the metabolism of which may be important in the development of bronchopulmonary pathology. Aim. To study the quantitative shifts in the spectrum of short-chain fatty acids in the condensate of exhaled air from smokers. Methods: Gas-liquid chromatography in the condensate of exhaled air was used to determine: C2 - acetic, C3 - propionic, C4 - oil, isoC4 - isobutyric, C5 - valeric, C6 - caproic, C7 - heptane acid. Results. Be level of acetate, butyrate, isobutyrate, valerate, heptane increases in the condensate of exhaled air of smokers. As the smoking experience and the number of cigarettes smoked per day increase, the concentration of exhaled volatile fatty acids increases, which may indicate metabolic, physiological disorders in the respiratory organs and, possibly, the beginning of the development of any pathological processes.
Problems of Biological Medical and Pharmaceutical Chemistry. 2019;22(4):43-46
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In vivo investgation of allergenic and immunotoxicity properties of redleaves extract isolated from Vitis vinifera L

Bortnikova V.V., Krepkova L.V., Babenko A.N., Borovkova M.V., Kuzina O.S.


Рurpose: complex assessment of the influence of grape culture (Vitis vinifera L.) leaves of red extract of dry on the immune system of experimental animals at the stage of preclinical studies. Methods: experiments were performed on albinos males guinea pigs (body weight 350-400 g), CBA and BALB/c males mice (body weight 18-20 g). Allergenicity was studied in tests of general systemic anaphylaxis (anaphylactic shock) and active skin anaphylaxis, as well as in delayed hypersensitivity reaction(HRT). Humoral immunity was assessed by the test of the generation of antibody-forming cells in the spleen by the method of local hemolysis in the modification Cunningham and the titer of hemagglutinins in the reaction of hemagglutination in the serum raised in microtitration Takachi. The state of T-cell immunity was investigated by induction test in the reaction HRT of CBA mice. Results: it was shown that the studied extract in the tested doses of 10 and 100 mg/kg and sensitization schemes did not cause a reaction of general anaphylaxis (anaphylactic shock) and active skin anaphylaxis carried out on albino guinea pigs. Grape leaf red extract dry, introduced with PAF, did not potentiate inflammation in the reaction HRT of BALB/c mice in the study of immunotoxic properties, it was found that the studied extract showed a stimulating dose-dependent effect on the primary humoral immune response and had no effect on the effectors of the cellular immunity of experimental animals. Conclusions: studies have shown that the grape leaf red extract dry did not have a sensitizing effect. The stimulating effect of the studied extract on T - dependent immune response of CBA mice was revealed.
Problems of Biological Medical and Pharmaceutical Chemistry. 2019;22(4):47-52
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Professor Oleg Nikiforovich Tolkachev (k 90-letiyu so dnya rozhdeniya)

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Problems of Biological Medical and Pharmaceutical Chemistry. 2019;22(4):53-54
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