The State of the Neuronal-Astrocytic Apparatus of the Hippocampus on the Background of Long-Term Administration of Isoniazid and Rifampicin under Conditions of Correction of the Gut Microflora

The purpose of the study was to determine the morphometric and ultrastructural features of neurons and glial cells, as well as the levels of glial fibrillar acid protein in the hippocampus of rats under long-term administration of isoniazid and rifampicin during experimental therapy with a combination of pro/prebiotics. Materials and methods. The studies were conducted on three groups of white Wistar male rats with drug-induced hepatitis, reproduced by intragastric administration of isoniazid and rifampicin for 28 days. In the last 14 days of the intragastric experiment, rats of the research group were administered probiotic and prebiotic. The content of cytosolic glial fibrillar acid protein in the hippocampus was determined by competitive enzyme-linked immunosorbent assay. Semi-thin sections of CA1 sections of the rat hippocampus were analyzed using the Image J. analysis program. The ultrastructural characteristics were studied using a PEM-100-01 transmission electron microscope (Selmi, Ukraine). Results and discussion. Long-term administration of isoniazid and rifampicin was associated with changes in the content of the cytosolic fraction of glial fibrillar acid protein obtained from the hippocampus. These changes were characterized by a tendency to increase the level of this protein by 17.8% (p=0.2) compared with intact animals. In contrast to the group of animals reproduced in the experiment drug-induced liver injury, in the experimental group, the level of glial fibrillar acid protein was significantly lower by 25.0% compared to the drug-induced liver injury group (p <0.05). Morphological analysis revealed a decrease in neuronal density in rats with drug-induced liver injury compared with intact control. At the same time, the specific number of degeneratively altered neurons in comparison with intact control in the group of drug-induced liver injury animals increased by 8.57 times. The number of degeneratively altered neurons in the group receiving yogurt / lactulose was 55.87 ± 4.23%, which was significantly higher than intact control rats, but was 19.4% (p <0.05) lower levels in rats with MIUP. Electron microscopic examination of rat hippocampal neuroglia was based on the study of changes in the astrocytic and oligodendroglial components. Changes in rat hippocampal neuroglia under experimental conditions were mainly related to the astrocytic link. Astrocytes were characterized by significantly lower cytoplasmic edema and adaptive-compensatory changes in the cell. Conclusion. Course administration of pro/prebiotics reduces the severity of manifestations of neurodegeneration, improves the state of astroglia in the hippocampus, and reduces the cytoplasmic levels of glial acidic fibrillar protein in animals with drug-induced liver injury

P–251 To collapse or not to collapse blastocysts before vitrification? A matched case-control study on single vitrified-warmed blastocyst transfers

Study question Does laser-induced artificial blastocoel collapse result in better blastocyst cryopreservation survival and higher live birth rate (LBR) in comparison with intact counterparts? Summary answer Compared to vitrification of intact blastocysts, collapsed blastocysts resulted in higher survival and for 5% higher LBR. Neonatal outcomes were comparable in both groups. What is known already Blastocysts have long been considered a stage that is suboptimal for freezing-thawing procedures due to their high fluid content and different cell types. The development of a modified vitrification technique has enabled blastocysts to better survive cryopreservation compared to a slow freezing procedure. Many studies on the optimization of cryopreservation of blastocysts have mentioned the need for artificial collapsing of the blastocoel prior to cryopreservation, thereby reducing the risk of intracellular ice-crystals formation. However, the effectiveness of artificial collapsing on blastocyst survival rate, single vitrified-warmed blastocyst transfer (SVBT) outcome and on safety of such intervention remains to be confirmed. Study design, size, duration A retrospective matched case-control study of transfers of single blastocysts being artificially collapsed (case) or intact (control) before vitrification. A sample size of 306 cycles in both arms was needed to achieve 80% power to detect a difference between the groups of 10% with P &lt; 0.05. Controls were matched to cases on a 1:1 ratio by female age, parity, fresh and frozen cycle protocol, blastocyst age and quality, getting 309 pairs of cases and controls. Participants/materials, setting, methods Artificial collapsing was introduced into clinical practice gradually. In fresh IVF cycles (performed in university clinic from 2012 until 2014) with supernumerary blastocysts, half of the blastocysts were randomly selected before vitrification for laser-induced artificial collapsing. The other half was vitrified in intact form. Only the first transfers of a single vitrified-warmed blastocyst (n = 818) were included in the study. By matching, 309 pairs of collapsed (study) and intact (control) SVBTs were identified. Main results and the role of chance Both groups were comparable by their characteristics in indications, female age, type and length of ovarian hyperstimulation, insemination method in fresh cycle, protocol for warmed blastocyst transfer, blastocyst quality and day of blastocyst vitrification. Survival rates in case and control group ((309/316) 97.8% and (309/323) 95.7%; P = 0.13) were comparable, but optimal survival rates (100% survival and re-expansion after warming) was significantly higher in artificial collapse group ((247/316) 78.2% and (225/323) 69.7%; P = 0.01). Clinical pregnancy rates ((120/309) 38.8% and (110/309) 35.6%; P = 0.4), miscarriage rates ((15/120) 12.5% and (24/110) 21.8%; P = 0.06) and LBR per transfer ((100/309) 32.4% and (85/309) 27.5%; P = 0.19) or LBR per warmed blastocyst ((100/316) 31.6% and (85/323) 26.3%; P = 0.14) were not statistically different between case and control groups. Since the study was powered to detect a 10% difference, the possibility of type 2 error cannot be excluded. Perinatal outcomes were available for 175 live births. There were 10.5% (10/95) preterm births in the study group vs. 16.3% (13/80) in control group (P &gt; 0.05). Birth weights (3,308 g (SD 592 g) vs 3,308 g (SD 738 g) and sex ratio (50.7% vs 49.2% boys) were also comparable between both groups (P &gt; 0.05). There were no major malformations detected in the study population. Limitations, reasons for caution The research is retrospective, but the cycles from both groups were performed in the same time period. The groups were balanced according to all possible confounders. Blastocysts for vitrification were first categorized by quality groups and embryos from each category were randomized for collapsing or for remaining intact. Wider implications of the findings: No significant difference was found in live births by this sample size. Nevertheless, increasing the success by 5% with the introduction of artificial collapsing can be an important step towards optimizing of blastocyst cryopreservation. To confirm a 5% improvement in results, a sample size of &gt; 2500 cases would be needed. Trial registration number The study has been approved by the National Ethics Committee of the Republic of Slovenia (0120–204/2016–2).

Effect of lysophosphatidic Acid on the Vascular Endothelial Growth Factor Expression in Autotransplanted Mouse Ovaries Encapsulated in Sodium Alginate

Objective: The aim of this study was to evaluate the effect of lysophosphatidic acid (LPA) supplementation during in vitro culture and transplantation of mouse ovaries on the follicular development and expression of vascular endothelial growth factor (VEGF) as an angiogenesis factor at the mRNA and protein levels. Materials and methods: Three weeks old mice ovaries were cultured in the presence and absence of LPA for 24 hours, then they were capsulated in sodium alginate in the presence and absence of LPA as four experimental groups. After transplantation the vaginal smears were performed daily to evaluate the initiation of the estrous cycle. The morphology and follicular distribution were analyzed at the first and fourth estrous cycles using hematoxylin and eosin staining. Then in the groups that showed higher and lower follicular development the immunohistochemistry assay was conducted to identify VEGF protein expression, and the real time RT-PCR was done to analyze the expression of Vegf gene at the first estrus cycle. Results: The large size follicles and also the corpus luteum were prominent in all transplanted groups at fourth estrus cycle in comparison with intact control groups. The statistically lowest percentage of small size follicles and the highest percentages of large size follicles were seen in LPA+/LPA- group (p<0.05). The expression ratio of Vegf to β-actin was significantly higher in this group in comparison with non-LPA treated and intact control groups (p <0.05). Conclusion: LPA as an angiogenesis factor increases the follicular development in transplanted ovaries but it causes early discharge of ovarian reserve.

Follicular development and the expression of BAX and vascular endothelial growth factor in transplanted ovaries in uni- and bilateral ovariectomized mice: An experimental study

Background: Several conflicting results have been reported on the survival and function of transplanted ovaries. Objective: Evaluation of the follicular development and the expression of vascular endothelial growth factor (VEGF) and Bcl-2-associated X protein (BAX) in ovaries transplanted into uni- and bilaterally ovariectomized mice. Materials and Methods: In this experimental study, 40 female NMRI mice (21-days-old, 12-15 gr) were ovariectomized uni- and bilaterally (n = 20/ group), while the 8-wk-old mice were considered as intact control group (n = 6). 5 weeks after transplantation at the proestrus stage, the morphology of recovered transplanted ovaries and the proportion of follicles were studied at different developmental stages. The apoptosis cell death by pro-apoptotic protein BAX and the expression of VEGF were evaluated using immunohistochemistry. Results: In the bilaterally ovariectomized mice, among the 455 counted normal follicles, a lower rate of primordial and primary follicles and a higher rate of preantral and antral follicles were observed (p = 0.002). However, the percentages of preantral and antral follicles, and the corpus luteum were significantly lower in the intact control group (among the 508 counted normal follicles in this group) compared to other transplanted groups (p = 0.002). The number of BAX-positive cells in all groups was not significantly different. The VEGF expression was prominent in vessels of the corpus luteum, and also in the theca layer of large follicles of studied groups. Conclusion: Early discharge of ovarian reserve was prominent in the bilaterally ovariectomized group but the incidence of apoptotic cells and VEGF expression as angiogenic factor did not differ in both ovariectomized mice. Thus, unilaterally ovariectomy has less side effects on the ovarian reserve compared to bilateral ovariectomy. Key words: Autotransplantation, BAX protein, Vascular endothelial growth factor, Ovariectomy, Mice.

Morphological assessment of changes in the cornea of experimental animals after ultraviolet corneal cross-linking

Purpose. To investigate the dynamics of morphological changes in the rat cornea after its ultraviolet cross-linking using various photosensitizer solutions. Material and methods. Studies were conducted on 20 rats in three groups. In the first group, 0.1% Riboflavin solution was used for saturation, in the second – 0.1% Riboflavin and 20% dextran; the third – intact (control). A UV cross-linking model (370 nm, 3 mW/cm2, 10 min) with corneal deepithelization with a diameter of 3 mm was used. Histomaterial processing was performed according to the standard procedure on the 3rd, 7th, and 30th days after surgery. Histological sections 1-4 microns thick were prepared on a rotary microtome RM 2145 (Leica Microsystems, Germany), stained with hematoxylin-eosin and using the van Gieson method. Light microscopy was performed using a LEICA DM 2500 microscope with a Leica DFC 450 digital camera (Leica Microsystems, Germany). Results. The main morphological changes in the rat cornea after UV cross-linking with 0.1% Riboflavin solution were associated with loosening of bundles of collagen fibers. This was caused by the development of stroma edema in the early postoperative period (3-7 days), which persisted up to 30 days of the experiment. A morphological feature of the use of Dextralink solution in corneal cross-linking is less pronounced stroma disfoliation, which is associated with the dehydrating effect of dextran. In both groups, a decrease in the density of stroma keratocytes was found. Key words: cornea, ultraviolet corneal cross-linking, corneal morphology.

Restoration of spermatogenesis in azoospermic mice by bone marrow mesenchymal stromal/stem cells conditioned medium

One of the main cause of male infertility is non-obstructive azoospermia, which is not manageable medically. The first aim of the current research was to show the effect of extracellular vesicle-contained conditioned media (CM) instead of mesenchymal stromal/stem cells (MSCs) for treatment of non-obstructive azoospermia. In the next step, we aimed to study the differentiation potential of MSCs into spermatocytes after injection of MSCs in mice seminiferous tubules. This study has provided an applied treatment for busulfan-induced azoospermia using adipose tissue-derived (AT-MSCs) and bone marrow-derived MSCs (BM-MSCs) and bone marrow CM (BMCM) in animal models. In this regard, 30 male adult Balb/C mice (30±5g) and two female eGFP+/+ Balb/C mice (30±5g) were used to design experimental groups and to culture stem cells, respectively. Then, six groups including intact control, azoospermia, AT-MSC therapy, BM-MSC therapy, BMCM therapy, and spontaneous healing groups were considered. All groups except intact control were induced azoospermia by injecting two doses of busulfan (10 mg/kg) with 21 days’ interval. Testes of all mice were removed and studied through histomorphometry and flow cytometry analysis 60 days after treatment. Histomorphometry and flow cytometry evaluation of testes showed normal morphology of most of the seminiferous tubules of therapy groups as well as successful recovery of spermatogenesis, but spermatogenesis was not observed in the azoospermia group. It is worth notable that the results of the BM-MSC therapy group were more favorable than other therapy groups. Consequently, AT-MSC, BM-MSC and BMCM can be strongly suggested as candidates in the therapy of azoospermia.

КОМПЛЕКСНОЕ СРЕДСТВО ВИРУКИЛЛ 260 ПРОТИВ ЯИЦ АСКАРИДИЙ У КУР Сафиуллин Р.Т., Чалышева Э.И., Краснобаев Ю.В.

КЛЮЧЕВЫЕ СЛОВА: ЦЫПЛЯТА, АСКАРИДИИ (ASCARIDIA GALLI), ЯЙЦА, КОПРОСКОПИЯ, СРЕДСТВА ДЕЗИНВАЗИИ, ВИРУКИЛЛ 260, ФЕНОЛ, ИНТЕНСЭФФЕКТИВНОСТЬ, ЭЙМЕРИИ АННОТАЦИЯ: Лабораторные исследования по изучению овоцидных свойств комплексного средства Вирукилл 260 в разных концентрациях в опыте in vitro на дистиллированной воде показали, что интенсэффективность препарата (доля погибших яиц аскаридий Ascaridia galli) составила: при концентрации 0,5% - 95,9%; 1 и 2% - 100%. Использованный в качестве препарата сравнения фенол (4%) в условиях лаборатории показал интенсэффективность 87,8%. Проведенная биопроба на цыплятах с 16 (инокуляция яйцами аскаридий) по 51 день жизни (возраст контрольного убоя) показала, что при неполном уничтожении аскаридий в случае использования фенола и 0,5% раствора Вирукилл 260 живая масса цыплят и ее среднесуточный прирост снижаются по сравнению с интактным контролем. Концентрация препарата 1% ранее показала себя эффективной также против спорулированных ооцист эймерий, поэтому ее рекомендуется использовать на практике для подготовки птичников к заселению молодняком для их дезинфекции против экзогенных стадий как аскаридий, так и эймерий. THE EFFICIENCY OF COMBINED PREPARATION VIRUKILL-260 AGAINST ASCARID OVA IN CHICKEN SAFIULLIN R.T.1, CHALYSHEVA E.I.1, KRASNOBAYEV YU.V.2 1 Federal Scientific Center “All-Russian Scientific Research Institute of Experimental Veterinary of K.I. Skryabin & Y.R. Kovalenko” of Russian Academy of Sciences 2 Ratsiovet Co The ovocidic properties of different concentrations of the combined preparation Virukill-260 in compare to phenolic solution were studied in vitro on the aqueous suspension of the ova of ascarids (Ascaridia galli). The efficiency of the elimination was 95.9% for 0.5% aqueous solution of the preparation, 100% for 1 and 2% solutions; for phenolic solution (4%) it was 87.8%. The in vivo test on four treatments of chicken (universal Yubileynaya breed, 5 birds per treatment) since 16 days of age (inoculation of treatments 2-4 with non-treated ova and ova preliminary treated with phenol and Virukill-260, 0.5%, respectively; intact control treatment 1 was not inoculated) to the slaughter at 51 days of age evidenced the decreases in live bodyweight and average daily weight gains in cases of incomplete elimination of the parasites in compare to the intact control. Since the concentration of the preparation 1% was earlier found to be effective against sporulated oocysts of the Eimerias it can be recommended for the disinfection of commercial poultry houses prior to the population for the complete preliminary elimination of ascarids and Eimerias at the exogenous stages of their life cycles. Keywords: CHICKEN, ASCARIDS (ASCARIDIA GALLI), OVA, COPROSCOPY, DISINVASIVE AGENTS, VIRUKILL-260, PHENOL, ELIMINATION EFFICIENCY, EIMERIAS

ЭФФЕКТИВНОСТЬ БИОТИНИЛИРОВАННОГО ПРОИЗВОДНОГО ОКИСЛЕННОГО ДЕКСТРАНА ПРИ ЭКСПЕРИМЕНТАЛЬНОМ ЭЙМЕРИОЗЕ КУР Коптев В.Ю., Афонюшкин В.Н., Давыдова Н.В., Онищенко И.С., Леонова М.А., Шкиль Н.А., Балыбина Н.Ю.

ИНФОРМАЦИЯ О ПУБЛИКАЦИИ Информационно-аналитическая система SCIENCE INDEX eLIBRARY ID: 44721476 DOI: 10.33845/0033-3239-2021-70-2-50-53 ЭФФЕКТИВНОСТЬ БИОТИНИЛИРОВАННОГО ПРОИЗВОДНОГО ОКИСЛЕННОГО ДЕКСТРАНА ПРИ ЭКСПЕРИМЕНТАЛЬНОМ ЭЙМЕРИОЗЕ КУР КОПТЕВ ВЯЧЕСЛАВ ЮРЬЕВИЧ1, АФОНЮШКИН ВАСИЛИЙ НИКОЛАЕВИЧ1, ДАВЫДОВА НАТАЛЬЯ ВИКТОРОВНА1, ОНИЩЕНКО ИРИНА СЕРГЕЕВНА1, ЛЕОНОВА МАРИНА АЛЕКСАНДРОВНА1, ШКИЛЬ НИКОЛАЙ АЛЕКСЕЕВИЧ1, БАЛЫБИНА НАТАЛЬЯ ЮРЬЕВНА1 1 Сибирский федеральный научный центр агробиотехнологий Российской академии наук (СФНЦА РАН), Новосибирская обл. Тип: статья в журнале - научная статья Язык: русский Номер: 2 Год: 2021 Страницы: 50-53 УДК: 576.852.24:575.827.2 ЖУРНАЛ: ПТИЦЕВОДСТВО Учредители: Министерство сельского хозяйства РФ ISSN: 0033-3239 КЛЮЧЕВЫЕ СЛОВА: КУРЫ, ЭЙМЕРИОЗ, БИОТИНИЛИРОВАННЫЙ ОКИСЛЕННЫЙ ДЕКСТРАН, ПАДЕЖ, МИКРОСТРУКТУРА КИШЕЧНЫХ ВОРСИНОК АННОТАЦИЯ: Изучена эффективность биотинилированного производного окисленного декстрана (БОД) для профилактики эймериоза кур. Цыплятам опытных групп вводили БОД с суточного возраста внутримышечно в дозе 0,02 мг/кг (группа 1) или орально в дозе 0,06 мг/кг (группа 2) один раз в 72 ч (5 введений); контрольной группе 3 (зараженный контроль) препарат не применяли. Цыплята этих групп на 10-й и 15-й день подвергались заражению смесью Еimeria acervulina и E. tenella. Группу 4 (интактный контроль) заражению эймериями не подвергали и препарат ей не применяли. Установлено, что максимальный падеж птицы после первичного заражения наблюдали в зараженном контроле - 37,5%. В опытных группах при внутримышечном введении БОД падеж составил 28,5%, при оральном - 14,2%. Максимальная средняя живая масса цыплят на 21 сутки опыта наблюдалась в группе 2, получавшей БОД орально; разница с показателем зараженного контроля составила 4,24%, интактного - 1,50% в пользу группы 2, что говорит о ростостимулирующем эффекте препарата. Средняя длина кишечных ворсинок в тощей кишке в группе 2 была достоверно больше на 37,7% по сравнению с зараженным контролем (р<0,001). Полученные результаты дают основание рекомендовать препарат БОД для профилактики эймериоза кур в дозе 0,06 мг/кг живой массы с первых дней жизни, орально каждые 72 ч (5 введений). THE EFFICIENCY OF A BIOTINYLATED DERIVATIVE OF OXIDIZED DEXTRAN IN CHICKEN WITH EXPERIMENTAL EIMERIOSIS KOPTEV V.YU.1, AFONYUSHKIN V.N.1, DAVYDOVA N.V.1, ONISHCHENKO I.S.1, LEONOVA M.A.1, SHKIL N.A.1, BALYBINA N.YU.1 1 Siberian Federal Scientific Center of Agrobiotechnologies of Russian Academy of Sciences The effects of a biotinylated derivative of oxidized dextran (BOD) in chicken with experimental eimeriosis were studied on 4 treatments of chicks of a layer cross (10 birds per treatment, 1-21 days of age). BOD was applied intramuscularly in the dose 0.02 mg/kg of bodyweight (treatment 1) or orally in the dose 0.06 mg/kg (treatment 2) since 1 day of age every 72 hours (5 applications totally); treatments 3 (infected control) and 4 (intact control) were not treated. Treatments 1, 2, and 3 were intragastrically infected by a mixture of Eimeria acervulina and E. tenella at 10 and 15 days of age. The mortality cases were recorded and diagnosed daily, average live bodyweight was recorded weekly; at 21 days of age all survived chicks were euthanized for microbiological and histological investigation of the intestine. Maximal mortality level after the first infection (37.5%) was found in treatment 3, in treatments 1 and 2 it was 28.5 and 14.2%, respectively. The highest live bodyweight at 21 days of age (83.33 g) was found in treatment 2, higher by 4.24% in compare to treatment 3 and by 1.50% in compare to treatment 4, evidencing the growth stimulating effect of the preparation. Average height of the jejunal villi in treatment 2 was significantly higher in compare to treatment 3 by 37.7% (p<0.001), the microscopy also evidenced the prevention of structural injuries in the intestinal villi induced by eimeriosis in treatment 3. The conclusion was made that BOD is an effective preventive preparation against eimeriosis in chicken, the most effective being five-fold oral administration since the first days of age at the dose 0.06 mg/kg every 72 hours. Keywords: CHICKEN, EIMERIOSIS, BIOTINYLATED DERIVATIVE OF OXIDIZED DEXTRAN, MORTALITY, HISTOLOGY OF INTESTINAL VILLI

Ultra structural Studies of Mouse Liver in Castrated Subjects Treated with Grape Juice

The present study was designed to evaluate the alterations in the liver tissues especially ITO and Kupffer cells of both castrated and castrated mice treated with10 µl/g of grape juice. The present study was conducted on forty five healthy males of Swiss albino mice, which were divided into 3 groups (N= 15 mice per each group). The first group was intact (control); the second one was castrated and the third group was castrated treated with 10 µl/g of grape juice. The ultra-structure sections of liver tissue from intact (control) group showed normal structure of both hepatocytes and sinusoids, with smooth external surfaces. Whereas ultra-sections from castrated group resulted in: degeneration of hepatocytes, deterioration of sinusoids; rough external surfaces, with aggregations of lipid droplets and white blood cells as compared with the control group. However treatment of castrated subject's with10µl/g of grape juice resulted in activation the regeneration processes in both hepatocytes and sinusoids, with smooth appearance of the external surfaces as compared with control group. The present study concluded that surgical castration aggravated increased hepatic steatosis and increased inflammatory response by increased activation of Kupffer and Ito cells, these effects could be reserved by aggravated increased grape juice administration.

EFFECT OF RHAPONTICUM CARTHAMOIDES AND CRANBERRY MEAL EXTRACTS COMPOSITION ON GLUCOSE LEVEL

It has been found that Rhaponticum carthamoides and cranberry meal extracts composition at an effective dose of 35:250 mg/kg after 14 days of oral administration to mice promotes a significant glucose level reduction on the background of physical and glucose challenge compared to intact control.