A method for the synthesis of a nanostructured turpentine oil lyosol by bubbling an emulsion of turpentine oil in water with air in a mass ratio of 1:200 to 1:2200 by the separation of nanoparticles using nanofiltration has been proposed. Changing the mass ratio of turpentine oil to the aqueous phase during the synthesis was shown to influence the size of the resulting nanoparticles. An increase in the Z-average size of nanoparticles from 50 to 320 nm along with a gradual decreasing turpentine content from 1:200 to 1:1000 was observed. A further decrease in the turpentine load from 1:1000 to 1:2200 led to decreasing the Z-average particle size up to 160 nm. The fact of the formation of stable non-covalent complexes of the lyosol particles with fluorescein has been established. The fluorescein-stained lyosol particles were used to study the efficiency of their phagocytosis by chicken blood monocytes using confocal microscopy. The maximum efficiency of the lyosol phagocytosis by the chicken monocytes was observed when the Z-average lyosol particle size was 50 nm, which was achieved with a turpentine dry matter ratio of 1:200 during the synthesis.

Курский государственный медицинский университет, Курск; В статье дается оценка современным данным по этиологии и патогенезу остеоартроза с акцентом на генетические механизмы развития данного заболевания.Анализ данных литературных источников позволяет высказать мысль, что суставной хондроцит является своеобразным датчиком гомеостаза суставного хряща и играет решающую роль в поддержании его нормальной физиологической структуры и функции.Гомеостаз суставных хондроцитов может быть нарушен в результате действия различных факторов, включая аномальную механическую нагрузку и старение, генетические нарушения

The article presents the haematological and biochemical blood parameters studies data obtained during the production test of the microencapsulated probiotic preparation Enzimsporin with an enzyme (crystalline trypsin) on Genesus genetics pigs in the fattening process. Data from three groups were analyzed: experimental group (microencapsulated Enzimsporin with enzyme), control group No. 1 (commercial non-encapsulated Enzimsporin) and control group No. 2 (without adding Enzimsporin to the feed). A general blood test showed that in animals receiving microencapsulated Enzimsporin with an enzyme, the erythrocytes and haemoglobin content reaches the upper physiological limit, which is significantly higher than in the control groups. When determining the blood biochemical parameters, a significant increase in the total protein and albumin amount in experimental animals was revealed by the 148th day of life. Reliably high glucose levels, within the physiological norm, were observed in animals receiving Enzimsporin, while in animals receiving the microencapsulated drug, the glucose level was significantly higher than in those receiving its unencapsulated form. The change in the trace elements level in the experimental animals’ blood showed regularities associated with an increase in the calcium amount and a decrease in phosphorus and magnesium. At the same time, an increase in the level of AST and ALT in the animals’ blood receiving Enzimsporin with an enzyme was noted by the 148th day of life. The observed immunoglobulins reliably high level throughout the experiment in experimental animals in comparison with control indicates a high resistance and pigs immune status.

The paper shows a comparative assessment of the effect of udenafil (Zidena®, Dong-A Pharmaceutical Co. Ltd., Korea) on the perfusion of healthy and ischemic tissues in critical ischemia of the lower extremities simulated in rats. Formed 4 experimental groups: 1 - intact; 2 - healthy animals with the introduction of udenafil (orally 8,6 mg/kg for 28 days); 3 - animals with ischemia without treatment; 4 - animals with ischemia and monotherapy with udenafil (similar to group 2). On the 21st and 28th days in all groups, the level of blood microcirculation in the leg muscles was recorded. At the same time, 10 animals were withdrawn from the experiment by anesthesia overdose and histological preparations of the leg muscles were prepared. The results obtained showed that udenafil increases regional blood flow both in non-ischemic tissues and in critical ischemia of the lower extremities.

Objective. So far, no experimental model of the necrotic ulcerative stage of critical limb ischemia has been created. Aim. Development of a method for modeling the necrotic ulcerative stage of critical lower limb ischemia.Materials and methods. The experimental study was carried out on 120 rats - males of the "Wistar" line, divided into 4 groups of 30 animals each. In the first group (intact), the level of microcirculation in the muscles of the hind limb was determined. In the second (sham-operated) group, ischemia was not simulated, but the neurovascular bundle of the thigh was isolated and the wound was sutured. In the third group (comparison), the modeling of chronic ischemia of the leg muscles was performed by removing the femoral, popliteal, anterior and posterior tibial arteries. In the fourth (experimental) group, to simulate chronic critical ischemia of the leg muscles, the femoral, popliteal, anterior and posterior tibial arteries and veins were removed, and the peripheral nerve was damaged.In the postoperative period, the dynamics of symptoms of limb ischemia was monitored. Animals were taken out of the experiment, 10 animals in each group by an overdose of anesthesia on the 10th, 21st and 28th days. Before hatching, the level of microcirculation was determined using laser Doppler flowmetry. To study the morphological picture at the same time in the third and fourth groups, the ischemic gastrocnemius muscle of the leg was excised and a standard histological examination was performed.Results. Trophic disorders in animals of the third group developed in 13.3%, and in the experimental group - in 100%. In the experimental group, the level of microcirculation at all periods was significantly less than in the comparison group: on the 10th and 21st by 1.3 times, on the 28th day by 1.2 times, and the area of ​​muscle necrosis was greater by the same days by 21.2%, by 8.2% and by 6.8%.Conclusion. The developed experimental model corresponds to the necrotic ulcerative stage of critical ischemia in humans.

The rationale for the study is explained by the fact that regular application of probiotic feeding supplements in pig breeding positively influences the growth and development of young stock, in particular, increases mean daily weight gain, normalizes the performance of the gastrointestinal tract and enforces the resistance of the organism. For this reason, a microencapsulated probiotic drug ―Enzymsporine withtrypsin‖ was developed in the facilities of Kursk State Agricultural Academy (Kursk, Russia). The authors studied the drug’s biological safety. The main method of the study of biological safety of the drug was the study of its subchronic toxicity that would allow for the production trials of this drug on agricultural animals. The article presents the experimental data obtained during the tests on Wistar rats.The tests on rats showed that new encapsulated probiotic drug ―Enzymsporine with trypsin‖ was safe in the doses that exceeded the recommended for the application in animal farming by 2 and 10 times. This provided perspectives for the application of this drug in animal production for the optimization of metabolism, an increase in the mass of animals, and the safety of meat products for humans.

The characteristics of recombinant phytase from Obesumbacterium proteus have been studied after its integration into Yarrowia lipolytica with a Po1f (pUV3-Op) plasmid. Phytase activity was observed in Y. lipolytica Po1f transformants in a wide pH range (3.5–7.5) and remained at 80% of the initial value after 3-min heating at 95°C, which made it possible to add the enzyme to a fodder produced with heat treatment. In the case of transformant cultivation on low-grade vegetable substrates, they also possessed confirmed phytase activity, as well as an increase in the content of intracellular phosphates as compared to cells of the wild-type Y. lipolytica strain.

Owing to the changes in the general ecological situation in Russia, livestock losses from ovine anaplasmosis have rapidly increased in recent years. The development of a vaccine against this disease is therefore all the more relevant. A continuous culture of tick cells could be used as an appropriate substrate for producing biomass of the pathogen of ovine anaplasmosis, Anaplasma ovis, for the purpose of manufacturing a cell-cultural inactivated vaccine. Such a continuous cell line of the tick Dermacentor marginatus, the definitive reservoir host of A. ovis, has been obtained by cultivation of homogenized tick eggs on a modified Leibovitz medium L15. The established cell line DM-77 has an undifferentiated phenotype: the cells are rounded, with large nuclei and well-visible nucleoli. By the end of the study, the cell line was passaged eight times and did not express signs of growth rate decrease. The duplication time was 2.8 ± 0.3 days for cultivation in a medium with 10% fetal serum at 32°C.

Неинвазивные дыхательные тесты с применением изотопно-меченых соединений представляют собой новый высокоточный и безопасный метод функционального исследования печени и билиарной системы. Целью работы было провести биологические испытания острой и субхронической токсичности 13С-меченых линолевой и линоленовой кислот, синтезированных по оригинальной методике и предназначенных для проведения диагностических дыхательных тестов. При однократном внутрижелудочном введении изучаемых соединений лабораторным мышам линии BALB/c и крысам Wistar в дозах, превышающих диагностические в 500–2500 раз, образцы соединений не вызывали смертности экспериментальных животных. При проведении субхронического эксперимента на крысах при дозировках испытываемых соединений, в 5 и 25 раз превышающих терапевтическую дозу для человека, в течение 14 суток было выявлено отсутствие достоверных изменений у животных в экспериментальных группах по сравнению с контрольной по массе тела, гематологическим показателям (содержанию эритроцитов, лейкоцитов и тромбоцитов в крови) и биохимическим показателям сыворотки крови (уровню гемоглобина, общего белка, щелочной фосфатазы, аланинаминотрансферазы, аспартатаминотрансферазы, лактатдегидрогеназы, билирубина). Исследованные меченые кислоты безвредны в дозах, планируемых для перорального введения, и могут быть рекомендованы к доклиническим и клиническим испытаниям.

Noninvasive stable isotope breath tests allow highly accurate and safe estimation of liver and biliary tract function. The aim of this study was to test 13С-labeled linoleic and linolenic acids intended for diagnostic use for acute and subchronic toxicity. The acids were synthesized using the patented method. A single intragastric administration of the tested compounds to experimental BALB/c mice and Wistar rats in the amounts exceeding clinical doses 500 to 2500-fold did not cause animal death. In the subchronic toxicity test, the rats received 5 to 25 times higher doses than recommended for clinical use in humans. In a 14-day follow-up period, no significant differences were observed between the main and the control groups in terms of weight, blood count (red blood cells, white blood cells, platelets), and blood biochemistry (hemoglobin, total protein, alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, bilirubin). The studied compounds are safe at doses intended for oral administration and are recommended for further preclinical and clinical trials.

ВЕСТНИК РГМУ 4, 2018 VESTNIKRGMU.RU | | Shevelev AB , Isakova EP, Trubnikova EV, La Porta N, Martens S, Medvedeva OA, Trubnikov DV, Akbaev RM, Biryukova YK, Zylkova MV, Lebedeva AA, Smirnova MS, Deryabina YI 1 Институт общей генетики имени Н. И. Вавилова РАН, Москва 2 Институт биохимии имени А. Н. Баха, ФИЦ Биотехнологии РАН, Москва 3 Курский государственный университет, Курск 4 Фонд имени Э. Маха, С. Микеле аль Адидже, Италия 5 Курский государственный медицинский университет, Курск 6 Курская государственная сельскохозяйственная академия имени И. И. Иванова, Курск 7 Московская государственная академия ветеринарной медицины и биотехнологии — МВА имени К. И. Скрябина, Москва 8 Федеральный научный центр исследований и разработки иммунобиологических препаратов имени М. П. Чумакова РАН, Москва

A new recombinant strain of Yarrowia lipolytica synthesizing encapsulated highly thermostable phytase of Obesumbacterium proteus, which is recommended for use as a premix component of feed compositions in animal husbandry, was obtained.

Management of purulent wounds is a problem that requires particular attention: wounds are a common injury type for which suppurative complications are frequent, mortality rates are high and antimicrobial therapy may be ineffective due to the presence of drug-resistant bacteria in the wound. In this work we have studied the effectiveness of wound treatment with the leaf extract of transgenic Kalanchoe pinnata modified to produce antimicrobial peptide cecropin P1. Purulent wounds infected with Staphylococcus aureus were modeled in Wistar rats. Four groups of animals were formed, with 10 animals in each group. In all groups, the wounds were cleansed with 3 % hydrogen peroxide solution once a day; all groups except the controls received additional treatment. Group 2 received 10 % cefazolin solution, group 3 received kalanchoe juice, group 4 received the juice of cecropin P1-producing kalanchoe. Histologic stains of biopsy samples were performed after rats were sacrificed by anesthetic overdose on days 3, 10 and 14 after treatment onset. On day 3, wound dynamics was the same in all groups. On day 10 exudate was still observed in the controls; in group two exudation was almost finished and regeneration was about to begin; in groups 3 and 4 the wound defect was filled with granulation tissue. In spite of epidermal repair along the wound edges in groups 2 and 3, there still was some sloughing and granulation tissue was less mature than in group 4. We recommend conducting more extensive clinical research of the leaf extract of cecropin P1-containing transgenic Kalanchoe pinnata.

Previously transgenic Kalanchoe pinnata plants producing an antimicrobial peptide cecropin P1 (CecP1) have been reported. Now we report biological testing K. pinnata extracts containing CecP1 as a candidate drug for treatment of wounds infected with Candida albicans. The drug constitutes the whole juice from K. pinnata leaves (not ethanol extract) sterilized with nanofiltration. A microbicide activity of CecP1 against an animal fungal pathogen in vivo was demonstrated for the first time. However, a favorable therapeutic effect of the transgenic K. pinnata extract was attributed to a synergism between the fungicide activity of CecP1 and wound healing (antiscar), revascularizing, and immunomodulating effect of natural biologically active components of K. pinnata. A commercial fungicide preparation clotrimazole eliminated C. albicans cells within infected wounds in rats with efficiency comparable to CecP1-enriched K. pinnata extract. But in contrast to K. pinnata extract, clotrimazole did not exhibit neither wound healing activity nor remodeling of the scar matrix. Taken together, our results allow assumption that CecP1-enriched K. pinnata extracts should be considered as a candidate drug for treatment of dermatomycoses, wounds infected with fungi, and bedsores.

In this work we explore the temporal dynamics of cytokines in Dark Agouti rats with experimentally induced autoimmune encephalomyelitis (EAE). The main group consisted of 11 animals who were injected with 100 mu l (per leg) of spinal cord homogenate obtained from random-bred rats and combined with incomplete Freund's adjuvant to the hind footpads. The control group included 7 animals who received 100 mu l of normal saline mixed with incomplete Freund's adjuvant. Blood samples (500 mu l) were collected daily, starting from day 1 through day 7. We ran a Bio-Plex-based multiplex cytokine assay on the samples using the Bio-Plex Pro Rat Cytokine 24-plex Assay kit. EAE in rats was shown to simulate progression of multiple sclerosis in humans in terms of temporal dynamics of lymphoproliferative and hematopoietic factors IL-1b, IL-2, IL-4, IL-5, IL-6, and IL-7. The studied model satisfactory imitates the dynamics of factors stimulating migration of lymphocytes, monocytes and other immune cells, including IL-17, RANTES (CCL-5) and MCP-1 (CCL-2) but excluding GRO/KC (CXCL1), which shows a different dynamics. The model also resembles patterns of human multiple sclerosis in terms of factors affecting cytotoxic and apoptotic reactions, including IFN gamma, IL-6 and IL-17, but excluding TNF alpha.

Procedure of manufacturing K. pinnata water extracts containing cecropin P1 (CecP1) from the formerly described transgenic plants is established. It included incubation of leaves at +4°C for 7 days, mechanical homogenization of leaves using water as extraction solvent, and heating at +70°C for inactivating plant enzymes. Yield of CecP1 (after heating and sterilizing filtration) was 0.3% of total protein in the extract. The water extract of K. pinnata + CecP1 exhibits favorable effect on healing of wounds infected with S. aureus (equal to Cefazolin) and with a combination of S. aureus with P. aeruginosa (better than Cefazolin). Wild-type K. pinnata extract exhibited evident microbicide activity against S. aureus with P. aeruginosa but it was substantially strengthened in K. pinnata + CecP1 extract. K. pinnata extracts (both wild-type and transgenic) did not exhibit general toxicity and accelerated wound recovery. Due to immunomodulating activity, wild-type K. pinnata extract accelerated granulation of the wound bed and marginal epithelialization even better than K. pinnata + CecP1 extract. Immunomodulating and microbicide activity of K. pinnata synergizes with microbicide activity of CecP1 accelerating elimination of bacteria.