Vol 23, No 3 (2020)

Follicle-stimulating hormone (FSH) is one of the most important hormones of reproductive system and it is widely used as a treatment in an assisted reproductive technology. Choice of productive cell line for recombinant human FSH (rhFSH) protein production is the major issue of biotechnological process. Study objective. The aim of the study was to generate stable cell lines for rhFSH production using different parental CHO cell lines and compare their characteristics. Material and methods. Two CHO suspension parental cell lines were used for stable cell line generation - CHO-K1 and CHO-DXB 11 adapted to suspension conditions. Cell count and viability were analyzed by trypan blue exclusion method. RhFSH concentration was measured by ELISA, protein identity was confirmed by western blot analysis. Results. Four clones generated from CHO-DXB 11 cell line were selected after 3 steps - pool selection, clone selection and methotrexate amplification. Productivity during 6 days of cultivation for the most productive clone CHO-FSH 11/24 was 7,1±0,6 mg/l. Three clones were chosen after transfection and cell cloning of CHO-K1 cells transfected with plasmids containing of FSH α- and β-chains sequences. The highest productivity was detected for CHO-FSH91 clone and it was 10,8±0,9 mg/l. Molecular weight of rhFSH was identical for rhFSH purified from cell culture of optimal clones and commercial preparation Gonal-F (Merck Serono).

The aim of the study was to develop a phage titer rise reaction (PRR) scheme for the indication of Xanthomonas campestris pv. campestris bacteria. Material and methods. The PRR scheme was developed using the bacteriophage Cl34-Ulgau and the indicator culture of X. campestris pv. campestris bacteria Xc2 is based on the methods applied by the staff of the Department of Microbiology of the Ulyanovsk State Agricultural University. Results. The paper presents the developed scheme of PRR. The optimum parameters for the formulation of the PRR. The working dilution of the phage biopreparation Kl34-ULSAU was 104 BOE/ml. The dilution of the indicator culture of bacteria 107 mc/ml. the cultivation time of crops with indicator culture and phage biopreparation was 18 hours. Conclusion. Identification of the bacteria X. campestris pv. campestris based on the developed phage titer rise reaction scheme is 43 hours.

The problem of male infertility especially affects HIV-infected men, so sperm motility decreases with highly active antiretroviral therapy. The aim of the study was to characterize the biochemical characteristics of sperm in HIV-infected men in order to restore their functional activity. Material and Methods. We observed 22 patients aged 25 to 45 years. This group has an undetectable viral load (40-50 copies / ml) and is receiving antiretroviral therapy. The studies were carried out using a Nikon Eclipse E 200 phase-contrast microscope. Determination of biochemical parameters of sperm plasma after standard sample preparation on a Roche Cobas integra 400+ automatic biochemical analyzer using Roche reagents and control materials. Results. The content of albumin, which determines the tonicity of the medium in the blood and the fulfillment of multiple vital functions, is almost 10 times lower than in blood plasma. It should be noted that the indicators of total protein and albumin in the sperm plasma of HIV-infected men are higher than in a healthy group of men and subfertile patients. The activity of aspartate-aminotransferase (ASAT) exceeds the blood plasma data. Moreover, the activity of ASAT is significantly higher in HIV-infected men. The activity of gamma-glutamyl transferase (GGT) is more than 200 times higher than in blood plasma. Further, as an experiment, we placed the "washed" spermatozoa in biological media (sperm plasma of a healthy donor, a woman's cervical fluid during ovulation) and obtained the following results. The percentage of progressively motile spermatozoa increased in the sperm plasma of healthy donors to 12%. We also observed an increase in sperm motility in the cervical fluid of a healthy woman during ovulation up to 7%. Conclusions. Thus, we have demonstrated that the use of sperm plasma from a healthy donor helps to restore sperm activity to 12%. The percentage of progressively motile spermatozoa increased in the sperm plasma of healthy donors. We observed an increase in sperm motility in the cervical fluid of a healthy woman during ovulation up to 6%.

Actuality. The fruits of Crataegus submollis Sarg. (family Rosaceae) are a popular food product, but are not currently used in medical practice. Previously, the possibility of using of the fresh fruits of Crataegus submollis Sarg. as a source of antidepressant and diuretic drugs has been shown. One of the new drugs may be a juice based on fresh fruits of Crataegus submollis Sarg. In еру fresh fruits of the Crataegus submollis Sarg. predominate the reduced forms of flavonoids, namely procyanidins. An earlier analysis of the juice from fresh fruits of Crataegus submollis Sarg. with the using of spectrophotometry showed that the maximum absorption in the UV spectrum is 282 nm, which coincides with the maximum absorption of the solution of catechin, a biogenetic precursor of procyanidins. The aim is the development of the method for quantitative determination of the total procyanidins in of the fresh fruits of Crataegus submollis Sarg. Material and methods. The object of the study was fresh fruits of Crataegus submollis Sarg. harvested in September 2019 on the territory of Samara and Samara region. The method of analysis was direct spectrophotometry at a wavelength of 282 nm calculated on catechin. Results. As a results of the study the method of quantitative determination of the total procyanidins (calculated on catechin) in fresh fruits of Crataegus submollis Sarg. by direct spectrophotometry at an analytical wavelength of 282 nm was developed. It was determined that the content of the total procyanidins in fresh fruits of Crataegus submollis Sarg. varies from 0,52±0,03% to 0,97±0,06% (calculated on catechin). The error of a single determination with a 95% confidence probability is ±5,81 %. Conclusion. The highest content of procyanidins can be noted in the sepals and the skin of еру fresh fruits of Crataegus submollis Sarg. The fresh fruits of Crataegus submollis Sarg. are promising medicinal plant raw materials.

The results of cultivation of Potentilla alba L. and Potentilla megalantha L. in vitro are presented. These species are valuable medicinal plants whose cells synthesize different classes of phenolic compounds, in particular tannins with high biological activity. Natural resourcesare not meeting the needs of the pharmaceutical industry. The search for alternative ways of reproduction in vitro of Potentilla plants and preservation of their biodiversity is an urgent task. The aim of this work was to study the reproduction characteristics of Potentilla alba L. and Potentilla megalantha L. in vitro. To achieve this goal, various methods of pretreatment of seeds were used: mechanical processing, heat treatment, soaking and stratification. As a result, of the conducted research, it was found that when introduced into the culture in vitro Potentilla alba L. and Potentilla megalantha L. it is necessary to apply seed stratification and carry out their germination on a hormone-free medium. Adding BAP to the nutrient medium at the concentration of 1 mg / l leads to the greatest increase in the biomass of P. megalantha and P.alba, as a result of the formation of the maximum number of adventive microbeads.