В рамках настоящего сообщения освещены современные научные проблемы, связанные с влиянием хронической нейрогенной боли и иных коморбидных патологий на развитие злокачественного процесса, а также экспериментальные разработки по митохондриальной терапии опухолей. Фундаментальные исследования проводили на новых моделях: 1. Бимодельная система роста меланомы В16/F10 на фоне хронической нейрогенной боли. Генерализация болевого синдрома способствовала отмене генетической программы сдерживания канцерогенеза в экспериментах на мышах с нокаутом по гену uPA. 2. Бимодельная система роста карциномы Герена и развития сахарного диабета у крыс. Сочетание патологических процессов привело к различным последствиям у животных разного пола: у самцов отмечалась стимуляция роста первичной опухоли, а у самок при умеренном росте первичного очага наблюдалась обширное метастатическое поражение брюшины, яичников и почек, что указывало на выраженное проонкогенное влияние диабета. 3. Бимодельная система роста карциномы Герена или саркомы 45 и гипотериоза, на фоне которого у крыс-самцов наблюдалось усиление роста опухоли и снижение продолжительности жизни по сравнению с самками. Это было обусловлено превалированием у самок эстрогенов, участвующих в регуляции работы щитовидной железы и оказывающих разное пролиферативное и антиапоптотическое влияние на клетки рака. Разработаны новые способы патогенетической митохондриальной терапии для предотвращения развития инфаркта миокарда и блокирования метастатической агрессии меланомы В16/F10 с эффектом замедления роста опухоли, растущей на фоне хронической нейрогенной боли у мышей обоего пола.

ОСОБЕННОСТИ РАЗВИТИЯ КАРЦИНОМЫ ЛЕГКОГО ЛЬЮИСА НА ФОНЕ ГИПЕРТИРЕОЗА У МЫШЕЙ ОБОЕГО ПОЛА Франциянц Е.М. 1 , Каплиева И.В. 1 , Васильева Е.О. 1 , Нескубина И.В. 1 , Сурикова Е.И. 1 , Трепитаки Л.К. 1 , Качесова П.С. 1 , Черярина Н.Д

e15054 Background: Previously, the antitumor efficacy of low intensity complex impacts, including electromagnetic radiation (EMR) of the extremely high frequency range (EHF) in combination with factors of electromagnetic or biochemical nature, was shown. The questions about systemic antitumor mechanisms and the role of the thyroid system in them are opened. The aim of the study was to evaluate changes in the levels of thyroid hormones and their ratio in the blood in the cases of tumor regression under the influence of complex impacts including EMR EHF. Methods: The experiments were carried out on outbred male rats of young (8-9 months) and senile (24-26 months) age with subcutaneously transplanted sarcoma 45 (S-45). EMR EHF (42.2 GHz, 10 mW/cm 2 ) with previously developed regime of modulation was applied on the head. In young males (21), the tumor was additionally exposed to the electric field with neuron-like impulses (SCENAR-97.1 device); in 20 old animals, complexes of essential L-amino acids were used per os 260 mg/kg/day. The impact course lasted for 3 weeks. After its completion, the levels of thyroxine (T4) and triiodothyronine (T3) in the blood were assessed by radioimmunoassay and the T3/T4 ratio was calculated. The results were compared with the values in intact animals (n=9-10), control group rats (S-45, n=11-15) and animals of the main groups without tumor regression. The Kruskal-Wallis and Mann-Whitney criteria were used, and the coefficient of variation (cv) was determined. Results: The tumor growth was accompanied by a decrease in T3 by 1.7-2 times (p <0.05) without changes in T4. The studied impacts caused a complete regression of S-45 in 38% (8) of young males, as well as a regression by 75-100% in 25% (5) of old animals. Young rats with S-45 regression showed no decrease in T3, while in old animals the T3 decrease was 1.3 times less pronounced than in the control group (p <0.05). In young males without S-45 regression, a decrease in T3 at the tendency level was observed (p <0.1), and T3 in old rats did not differ from the control group. The level of T4 did not change. In all cases of S-45 regression, despite the variability of T3 and T4 separately (cv=31-68%), practically constant value of the T3/T4 ratio was noted (cv=2-4%). It was 16x10 -5 in young animals and 13x10 -5 in old ones. It differed by 12-94% from the T3/T4 in the comparison groups and in rats without tumor regression (p <0.05-0.01). In the latter cases, the T3/T4 ratio had significant variability (cv = 25-84%) and was outside the range of values in rats with tumor regression. Conclusions: The results indicate the participation of the thyroid gland in the effective mobilization of systemic antitumor mechanisms, the marker of which is the ratio of the thyroid hormone levels in the blood.

e21559 Background: Gender differences in brain physiology and gender differences in pathology are usually recognized, but often are disregarded in clinical and experimental studies, resulting in numerous inaccuracies in data interpretation. The purpose of this study was to analyze levels of neurosteroid hormones in mitochondria of the cerebral cortex cells in C57BL/6 mice with subcutaneous B16/F10 melanoma growing in presence of chronic neurogenic pain (CNP). Methods: The study included male and female C57BL/6 mice (n = 336) aged 8 weeks initially weighing 21-22 g. Experimental groups were: intact animals; controls with a CNP model created by bilateral sciatic nerve ligation; comparison group 3 weeks after subcutaneous inoculation of 0.5 ml suspension of B16/F10 melanoma cells diluted 1:10; main group 3 weeks after subcutaneous melanoma growth in presence of CNP (CNP+B16/F10). Levels of estradiol (pg/g protein), estrone (pg/g protein) (DBC, Canada); progesterone (ng/g protein), total and free testosterone (pg/g protein) (XEMA, Russia) were measured in mitochondrial samples by ELISA (Tecan Infinite F50 analyzer, Austria). Results: Levels of estradiol in intact females were 3.1 times higher than in males, while estrone, progesterone, total and free testosterone were lower by 6.4, 2.7, 2.0 and 2.5 times, respectively. Only in females with CNP estradiol decreased by 3.4 times, compared to intact values, and estrone increased by 1.7 times (p<0.05), testosterone by 3.9 times. Estradiol in animals with B16/F10 decreased compared with intact values by 3.1 times in females and by 1.5 times in males (p<0.05). Females with B16/F10 showed the highest levels of progesterone exceeding intact values by 3.1 times. In the group with CNP+B16/F10, females showed lower levels of estradiol and estrone (by 1.4 times, p<0.05) and free testosterone (by 1.6 times, p<0.05), compared to the levels in CNP; on the opposite, males had 1.5 times (p<0.05) higher estrone and 2.9 times lower progesterone. Conclusions: Low levels of estradiol involved in the protective mechanism of neurosteroids were the dominant factor in mitochondria of cerebral cortex cells in females with CNP and malignant neoplasms. Males did not demonstrate such dominant factor. In animals with CNP+B16/F10, the response nature of cerebral cortex mitochondria changed: all defense mechanisms in the brain of females were suppressed by the 3rd week of tumor development in presence of pain. In males, mitochondria of cerebral cortex cells were more resistant to the influence of two pathologies, and only few changes in the neurosteroid status were recorded.

Mitochondria are dynamic organelles which constantly change their shape, size, and location within the cells. Mitochondrial dynamics is associated with mesenchymal metabolism or epithelial-mesenchymal transition to regulate the stem cell differentiation, proliferation, migration, and apoptosis. The transfer of mitochondria from one cell to another is necessary to improve and maintain homeostasis in an organism. Mitochondrial transplantation is a therapeutic approach that involves an introduction of healthy mitochondria into damaged organs. Recent evidence data have shown that the physiological properties of healthy mitochondria provide their ability to replace damaged mitochondria, with suggesting that replacing damaged mitochondria with healthy mitochondria may protect cells from further damage. Moreover, mitochondria can also be actively released into the extracellular space and potentially be transferred between the cells in the central nervous system. This increased interest in mitochondrial therapy calls for a deeper understanding of the mechanisms, which build the basis for mitochondrial transfer, uptake, and cellular defense. In this review, questions related to the involvement of mitochondria in the pathogenesis of cancer will be discussed. Particular attention will be paid to mitochondrial transplantation as a therapeutic approach to treat the mitochondrial dysfunction under some pathological conditions.

e23556 Background: The system of insulin-like growth factors (IGF) is involved in the pathogenesis of many malignant tumors. At the same time, the specificity of this system in recurrent soft tissue sarcomas is unknown. The purpose of this study was to analyze the IGF system in tumors and their peritumoral areas in men with recurrent soft tissue sarcomas. Methods: The study included 26 men aged 57.8±6.2 years with primary (n = 12, group 1, controls) and recurrent (n = 14, group 2, main group) soft tissue sarcoma of the extremities, T2bN0M0. Grade 1 tumors were diagnosed in 6 patients of group 1 and in 7 patients of group 2, while the other patients had G3 or G4 tumors. 95% of tumors were liposarcomas. All patients of group 2 had previously underwent surgical and radiation treatment for primary sarcomas and their relapses (up to 2 episodes), with the last surgery more than 1 year ago. Levels of the IGF system components were determined in tumor (T) and peritumoral (PT) tissues by ELISA, and ratios of IGF(T)/IGF(PT) were calculated. Results: Levels of the IGF system components in patients of group 1 were similar in T and PT. In group 2, levels of IGF1 and IGFВР2 in T were respectively 2.2 and 1.6 times (p < 0.05) lower than in PT, but did not differ significantly from the levels in group 1. The IGF1(T)/IGF1(PT) ratio in patients of group 2 was 1.8 times (p < 0.05) lower than in group 1. PT levels of IGF2 in group 2 depended on the tumor grade: they were 1.5 times (p < 0.05) higher in G1 than in G3-G4, while no such dependence was observed in T. Conclusions: The levels of the IGF system components in recurrent soft tissue sarcomas in older men had some peculiarities. Unlike primary sarcomas with similar levels of the studied components of the IGF system in tumors and in peritumoral tissues, recurrent tumors contained less IGF1 and IGFBP2 than the corresponding peritumoral areas. The levels of IGF2 in the peritumoral tissues of recurrent tumors depended on the tumor grade.

Aims are to study the nature of the processes of carcinogenesis of experimental B16/F10 melanoma in uPA gene knockout mice modified by chronic neurogenic pain and investigate some electrophysiological mechanisms of melanoma development. Materials and methods. We used 48 C57BL/6-PlautmI. IBug-ThisPlau6FDhu/GFDhu mice of both genders with urokinase gene knockout and 102 C57BL/6 mice of both genders with the normal genotype. Chronic neurogenic pain (CNP) was produced due to bilateral ligation of the sciatic nerve. Against the above background, all animals were transplanted with B16/F10 melanoma. To study the mechanism of CNP, studies of the intracellular electrophysiological activity of neurons of the central nervous system of the snail Helix pomatia in the body in vivo were carried out. CNP was reproduced by dosed pressing of four main nerves with Fresnel hairs that with time turned into increasing pain. Membrane potential (MP), action potential (AP) and firing rate (FR) parameters of intracellular bio-potentials of the command neuron RPaG3, continuously recorded using an ultrathin glass microelectrode for 4-5 days, were analyzed. Results. It was detected that an activation of cancerogenesis during the modification of the progression of experimental B16/ F10 melanoma in C57BL/6-PlautmI.IBug-ThisPlau6FDhu/GFDhu mice with uPA gene knockout using CNP is accompanied by a 2-fold acceleration in the time of tumor production, stimulation of the growth of the primary tumor nodes from 1.05±0.08 cm3 to 9.50±0.98 cm3 (p<0.001) and multiple metastasizing to the lungs, a reduction in the life span from 34.67±0.67 to 21.33±2.19 days (p<0.001) <0.05) in the genetically modified mice, by changing some gender-specific characteristics of the progression of the malignant process. The neuropathic nature of pain resulting from command neuron compression or ligation of the sciatic nerves is essentially identical to the implementation of genetic programs responsible for the control of life and death, reproducing events in carcinogenesis with the progression of a malignant tumor. Conclusion. The initiation and chronization of pain at the local level of the nervous system can lead to generalization of the pain syndrome and contribute to the cancellation of genetically predetermined programs of carcinogenesis.

11556 Background: Tumor necrosis factor alpha (TNFa) and its receptor (TNFa-R) play an important role in tumor genesis. However, their involvement in the recurrence of sarcomas is poorly studied. The purpose of this study was to analyze levels of TNFa and TNFa-R in the blood of patients with recurrent soft tissue sarcomas. Methods: The study included 64 male and female patients, mean age 63.4±5.2 years. Main groups included patients with recurrent G1 and G3 soft tissue sarcomas T2bN0M0; comparison groups included patients with primary G1 and G3 soft tissue sarcomas T2bN0M0. 95% of tumors were liposarcomas, with solitary rhabdamyosarcomas, myxofibrosarcomas, epithelioid and undifferentiated sarcomas. All recurrent patients had previously underwent surgical and radiation treatment for primary sarcomas and their relapses (up to 2 episodes), with the last surgery more than 1 year ago. Control groups (n = 10 each) included healthy donors of similar age. Levels of TNFa and TNFa-R were measured in the blood serum by ELISA before the treatment. Results: No significant differences were found between TNFa levels in patients with primary and recurrent soft tissue sarcomas and in donors. Levels of TNFa-R in men with primary G3 sarcomas were 1.5 times (p < 0.05) higher compared with donors, 1.7 times (p < 0.05) higher than in men with primary G1 sarcomas, and 1.3 times (p < 0.05) higher than in women with primary G3 sarcomas. TNFa-R increased by 1.7 times (p < 0.05) in all men with recurrent sarcomas regardless of the tumor grade, compared with the levels in healthy men; however, in G1 it was 1.9 times (p < 0.05) higher than in men with primary G1 sarcomas, while in G3 it did not differ significantly from the levels in men with primary G3 sarcomas. TNFa-R in women increased only in recurrent G3 sarcomas by 2.5 times compared to healthy women. No differences were observed in TNFa-R levels between men and women with recurrent G3 sarcomas. Conclusions: An excess of TNFa-R in the blood is characteristic for the recurrent soft tissue sarcomas G1 in older men and G3 in older women. At the same time, high grade tumors in men are accompanied by an increase in TNFa-R in the blood in both primary and recurrent sarcomas.

The aim of this research work is to study the cAMP level in the cardiac mitochondria and serum in the C57BL/6 strain mice of both genders under the independent melanoma B16/F10 growth versus the melanoma B16/F10 growth linked to chronic neurogenic pain (CNP). Materials and methods. Mice of strain C57BL/6 (n=336) have been grouped as follows: the intact group of the mice (male n=21; female n=21), the reference group (male n=21; female n=21) with the reproduced CNP model, the comparison group (male n=63; female n=63) to include the mice with melanoma B16/F10, and the main test group (male n=63; female n=63) to cover the mice with the melanoma growth against the CNP background. Upon expiration of 1 week, 2 and 3 weeks of the melanoma growth, in the animals of the above experimental groups the cardiac mitochondria have been isolated with the centrifugation using high-performance refrigerated centrifuge Avanti J-E, BECMAN COULTER, USA. With ELISA Kit (RayBio USA) we have determined cAMP concentrations in serum and in the cardiac mitochondria. Results. CNP has induced a decrease in the cAMP level in the cardiac mitochondria by a factor of 3,6 in the female mice only. In the animals of the comparison group the cAMP level in the heart has been increasing beginning with week 2 of the tumor growth on average by a factor of 4, while in the main test group starting from week 1 of the tumor growth it has been recorded 2-4 times higher and was depleted by the end of the experiment. As to the cAMP concentration in serum, the dynamics thereof has not been found to be in correlation with the cardiac mitochondrial data, and its concentration decrease has been recorded both in the females and the males. Conclusion. So, the changes in the cAMP concentration in the cardiac mitochondria demonstrate their gender-specific feature; the female mice as against the males have responded to an independent impact produced by CNP. As to the main test group, CNP has stimulated an increase in the cAMP level in the cardiac mitochondria 1 week earlier than it is the case with the comparison group, and it has resulted in the full cAMP depletion by the 3rd week of the experiment.

To study the intensity of lipid peroxidation (LPO) and the activity of the main antioxidant protection enzyme: superoxide dismutase (SOD) in heart tissues and tumors in rats of both genders with Guerin’s carcinoma (GC) and the tumor growth against the background of diabetes mellitus (DM). Materials and methods. Our research work was carried out in 80 outbread albino male and female rats, divided into 4 groups, with 10 animals of each gender in a group. The animals of two groups, namely, an intact animal group and a group of rats treated with alloxan DM (with a 5-fold increase in glucose levels) were subcutaneously transplanted with the Guerin’s carcinoma (GC) strain cells, and at the same time we used one group of the intact rats and another group of the animals with DM as the references. The content of malondialdehyde (MDA), diene conjugates (DC), and the SOD activity in the heart and the tumor tissues were determined by conventional spectrophotometric methods. Results. The most pronounced changes were found in the heart in the female rats with isolated GC and GC growing against the background of DM: a more than threefold increase in MDA, with a significant increase in DC and a multiple increase in the SOD activity as compared with the intact animals. In the GC tissue, the dependence of the severity of the increase in the MDA content on the size of the tumor was traced: the maximum increase in both parameters was observed in males with GC tumor growing against the background of DM. In the males, the volume of the subcutaneous tumor nodes was 1.8 times greater than that in the reference group and in the females with combined pathology, while in the females with combined pathology the volume of their tumors was 1.3 times less than that in the reference group, although the area of tumor lesions in them was maximum due to extensive metastasizing. Conclusion. DM has changed the specifics of oncogenesis depending on the gender of the animals. The identified gender differences in the redox status of the heart and the tumor in rats with combined pathology contribute to specifics of oncogenesis in males and females and determines their life expectancy.

e17515 Background: The development of deep postovariectomy disorders in young women with cervical and breast cancers is of extreme concern. They are associated with the suppression of the hypothalamic-pituitary-gonadal axis of regulation, estrogen involution, a sharp depression of the psycho-emotional state, and social distancing. In fact, these postovariectomy syndrome (POES) events are associated with low-reactivity stress syndrome. Our purpose included the transition of dominant stress into archetypes of anti-stress adaptive reactions under the influence of programmable exponential dose regimens of xenon-oxygen therapy (XOT) in the early period after the removal of the female reproductive organs. Methods: 123 patients of reproductive age diagnosed with cervical cancer pT1B2N0M0 and 24 patients with hormone-positive breast cancer pT2N1M0 and concomitant gynecological pathology underwent hysterectomy with oophorectomy and developed POES. All patients received a cycle (5 procedures) of low-dose inhalation XOT. The therapy consisted in a gradual increase in the percentage of xenon in the inhaled xenon-oxygen mixture with a reciprocal decrease in exposure time, an exponential regimen in the concentration range from 12–14% to 20–24% and exposure from 25 to 10 minutes. Blood levels of FSH, LH, estradiol, progesterone, and testosterone were measured by RIA (Immunotech, Czech Republic) before and after XOT. The psycho-emotional status was assessed by the generally accepted quality of life scales for cancer patients MOS-SF-36 and ESAS, the type of adaptive reactions was identified by Garkavi using the analysis of Schilling's leukogram, the severity of POES was determined by the menopausal index (MMI). Results: The POES manifestation and the dominance of acute stress were due to the inversion of hormonal metabolism - a decrease in the level of estradiol 487.3±52.4 and progesterone 1.8±0.1 relative to the initial levels 1017.9±83.4 nmol/L and 11.8±0.7 nmol/L, respectively, with an increase in FSH by 5.8 times, and LH by 2.4 times (p < 0.05). An accompanying XOT resulted in a significant MMI decrease (p < 0.05), training and calm activation reactions prevailed in 80% cases with harmonious changes in the regulation of hormonal processes and a clear regression of the acute estrogen deficiency (elevation of estradiol levels to 751.4±61.4 nmol/L and a decrease in FSH and LH by 1.5 and 2.1 times, respectively (p < 0.05)). MOS-SF-36 and ESAS assessment showed a significant decrease in pathological symptoms. Conclusions: XOT in the early postoperative period in cancer patients of the reproductive age with POES normalized their hormonal status, corrected functional disorders and improved their quality of life.

e23558 Background: Experimental data showed proliferative and anti-apoptotic effects of thyroxine (T4) and triiodothyronine (T3) on cancer cells which regulate gene expression and stimulate estrogen-like effects. The purpose of the study was to analyze effects of hypothyroidism on the development of malignant tumors in male and female rats. Methods: Female (n = 15) and male (n = 15) white outbred rats weighing 150 g and more received mercazolil (2.5 mg/100 g of weight) daily for 30 days (total dose of 75 mg/100 g of weight). Blood levels of T3, T4 and TSH were measured in all animals. When persistent hypothyroidism was observed, Guerin's carcinoma (GC, group 1) and sarcoma 45 (S45, group 2) were inoculated at a dose of 2 million cells in 0.5 ml of saline under the skin of the back to animals of both genders. The control group included rats of both genders with subcutaneous inoculation of Guerin's carcinoma and sarcoma-45 in the same dosage and volume but without preliminary reproduction of the hypothyroidism model. Results: Average tumor volumes in females with GC and hypothyroidism were less than in animals of the control group: after 4 days by 1.3 times (p < 0.05), after 7 and 10 days by 1.4 times (p < 0.05), after 14 days by 1.5 times (p < 0.05), after 18 days by 1.3 times (p < 0.05), and after 21 days by 1.4 times (p < 0.05). The survival of female rats in the main group was 1.6 times higher (p < 0.05) than in rats of the control group. Average tumor volumes in females with S45 and hypothyroidism were less than in animals of the control group: after 4 days by 1.4 times (p < 0.05), after 7 and 10 days by 1.6 and 3.2 times (p < 0.05), after 14 days by 3.9 times, and after 18 days by 4.8 times. The survival of female rats in the main group was 1.8 times higher (p < 0.05) than in rats of the control group. Average tumor volumes in males of the main group with GC and hypothyroidism after 18 and 21 days were similar to the values in animals of the control group. Their survival did not differ from the survival of control males. Average tumor volumes in males of the main group with S45 and hypothyroidism after 10 and 21 days were similar to the values in animals of the control group. Their survival did not differ from the survival of control males. Conclusions: Hypothyroidism in female rats with GC and S45 inhibited the growth of malignant tumors and improved the survival of animals, while in males no such inhibiting effect was observed.

e17554 Background: Thrombosis in patients with gynecologic cancers worsens the outcome of antitumor treatment and is one of the leading causes of their death. The kallikrein-kinin system (KKS) is involved in both the regulation of thrombus formation and the development of cancer. The purpose of this study was to analyze characteristics of the KKS components in the blood in patients with endometrial cancer (EM) and ovarian cancer (OC) with and without secondary thrombosis (VTEC). Methods: The study included 39 patients, mean age 58.0±4.2 years: main groups – patients with OC T1c-3cN0M0 (n = 10) or EC T1a-2N0M0 (n = 9) with VTEC; comparison groups - patients with OC (n = 10) or EC (n = 10) without VTEC. EC was represented by adenocarcinomas (G1-G3), OC by serous carcinomas (90%) and clear cell adenocarcinomas (10%). The control group included healthy women of the corresponding age (n = 10). Blood levels of kallikrein 1 (K1), kallikrein 14 (K14), and kininogen (KG) were measured by ELISA after the surgery. Results: EC patients with VTEC showed 1.5 times (p < 0.05) higher blood levels of K1, compared to donors, while other indices were unchanged. EC patients without VTEC had 4 times lower KG levels, compared to donors and EC+VTEC. In OC, regardless of the presence or absence of VTEC, levels of K1 in the blood increased, as well as in women with EC+VTEC, by 1.5 times (p < 0.05) compared with donors, which was combined with a twofold increase in KG in OC+VTEC and a decrease in KG by 1.4 times in OC patients without VTEC. Blood levels of K14 increased only in OC patients without VTEC by an average of 1.9 times (p < 0.05) compared with donors and OC+VTEC. Conclusions: The revealed changes in some KKS components in the blood demonstrate the similarity (K1 increase) and differences (KG increase only in OC) in the pathogenesis of thrombosis associated with gynecologic cancers. The mechanisms of protection against VTEC in the early postoperative period also showed common (KG decrease) and specific features associated with the characteristics of cancer (K14increase only in OC). The development of therapy aimed at correcting the identified disorders will allow an antitumor treatment program for this category of patients with maximum efficiency improving their quality of life and survival.

Topicality. An increase in the incidence of malignant tumors progressing against the background of various comorbid pathologies determines the need to study the mutual influence of pathological processes using experimental modeling. Such models, for example, can reproduce the tumor growth modified by the comorbid condition of diabetes mellitus, the incidence rate of which is now increasing exponentially. In this case, a clear demonstration of changes in the structure of organs outside the zone of the primary tumor node, using data on the direct growth of the tumor and the content of diabetes markers therein and in the perifocal zone, can serve as an evidence-based argument for the implementation of the mechanism of modified tumor progression. The aim of our research work is to assess the state of the histological structure of some internal organs (the kidneys, the ovaries, the peritoneum) when modeling the main pathological process in animals, the growth of Guerin’s carcinoma, against the background of a comorbid state of experimental diabetes. Materials and methods. We used 32 outbred male and female rats weighing 180-220 g to reproduce the model of experimental diabetes by a single intraperitoneal injection of alloxan at a dosage of 150 mg/kg of body weight. One week after the production of persistent hyperglycemia in the range of 25.4±1.2 mmol/l, the animals were transplanted with Guerin’s carcinoma subcutaneously in the region of the right shoulder blade. Upon expiration of 2 weeks, the animals were decapitated, and the harvested organs were prepared according to the practice stages of morphological preparation for staining sections with hematoxylin-eosin, followed by morphological examination of the structure with the use of the Leica DM LS2 microscope with the Olympus optical. C-5050 Zoom video camera and the Morfotest software. Photographing was carried out with magnification x10, x40, x100. Results. Our study of the morphological portrayal of the ovary, the kidney, the visceral and parietal peritoneum bears witness to the identity of the changes, consisting in a total metastatic lesion and abnormal transformation of the normal structure of all the studied organs only in the female rats, when modeling the comorbid state of diabetes mellitus. At the same time, the aggressive nature of the tumor progression was manifested in the blood filling of the vessels and hemorrhage, followed by the release of tumor cells, their settlement, the enhanced proliferation, the formation of strands and compaction of cell aggregations throughout the volume of the organ. Some gender specific features of the tumor progression were noted, which were found in the female rats along the path of active metastasizing in case of small primary tumors, and in the male rats along the path of stimulating the growth of the primary focus without metastasizing. It was revealed that these differences are associated with different degrees of saturation of the tumor and perifocal zone with glucose, and they are determined by the state of the insulin/insulin-like growth factor (IGF) axis. Conclusion. Morphological examination of the organs affected by metastatic Guerin’s carcinoma in the female rats with diabetes mellitus makes it possible to detect not only the synergy of both pathological processes, but also a powerful pro-oncogenic effect of the comorbid state of diabetes in the implementation of the tumor growth program.

The aim is to evaluate the physiological parameters of the efficacy of cardiac mitochondria transplantation in male mice with chronic neurogenic pain and B16/F10 melanoma growth. Materials and methods. Male mice (n=37) of the C57BL/6 line were used in the research work. The animals covered experimental groups as follows: mice with chronic neurogenic pain (CNP) + B16/F10 melanoma (n=27); mice with CNB + B16/ F10 melanoma + mitochondrial therapy (MC therapy) (n=10). Mitochondria were isolated from the heart of an intact rat with the use of differential centrifugation. An introduction of mitochondria to mice was carried out daily intraperitoneally at a dose of 3.3 mg of protein for 3 weeks. Statistical analysis of the results is carried out with the Statistica 10.0 software. Results. On day 21 (week 3) of the experiment, macroscopically in the melanoma tissue in the group of animals with MC therapy, there were 2.5 times more necrosis cases than in the group without MC therapy. During the examination of the internal organs, no metastases were detected in the animals treated with MC therapy, while in 100% of the animals without MC therapy metastases were found in the lungs and in 95% of them in the spleen. In the animals received MC therapy, there was no damage to the heart muscle in 75% of the cases, while in the group of the animals without MC therapy, the presence of lesions in the form of bruises on the surface of the heart was macroscopically detected in 100% of the animals. Conclusion. Thus, intraperitoneal transplantation of intact heart mitochondria contributed to the prevention of myocardial infarction and metastases to internal organs in the C57BL/6 male mice with B16/F10 melanoma growing against the background of chronic neurogenic pain.

Abstract Regenerative medicine is a vital area that advances methods addressing the process of liver regeneration. The liver has the unique ability to return to standard size within a short period of time after its injury or partial hepatectomy. Hepatocytes are highly active metabolic cells, which have a large number of mitochondria, suggesting a biologically significant role for the mitochondrial dynamics in hepatocyte damage and repair. The aim hereof is to develop a method for accelerating the process of liver regeneration in rats based on the transplantation of mitochondria isolated from the liver of intact rats. Materials and methods. Experimental animals (n = 30) used by us were white outbred rats divided into groups as follows: 1) a comparison group to include intact animals (n = 10); 2) a reference group with liver resection + use of saline solution (n = 10); 3) the main group with liver resection + mitochondrial therapy (MCT) (n = 10). In the reference and the main groups, under xylazoletil anesthesia, median laparotomy was performed and the maximum allowable resection of the left lobe of the liver was executed. Mitochondria of the intact liver were diluted with a 0.9% NaCl solution to a protein concentration of 11.6 g/l and injected into the post-surgery rats of the main group (group No. 3). The above procedure was carried out for 14 days. Animals of the reference group were intraperitoneally injected with 0.5 ml of saline solution according to a similar scheme. Statistical analysis was performed with the Statistica 10.0 Software. Results. According to the weight indices of the liver after MCT, the mass of the organ increased by a factor of 1.7 р˂0.05) relative to the reference group without MCT. At the same time, compared with the indicators in animals of the comparison group (intact animals), the mass of the organ exceeded 1.6 times (p˂0.05), and the liver stump itself during MCT in its mass exceeded by 3.2 times the weight indicators of the liver stump in rats without the MCT. Conclusion. As a result of the experiment on the use of MCT of the liver, it can be concluded that the resection of the liver lobe and subsequent processes in the liver tissue against the background of MCT acquire pronounced features of accelerating the physiological regeneration, addressing the restoration of stromal and parenchymal components.

e21561 Background: Global trials form a new mitochondrial paradigm for suppressing the growth of malignant tumors. The unique role of mitochondria (MC) in the processes of metabolism, proliferation, and cell death was proven by the establishment of signaling pathways and critical conditions for the self-organization and degradation of MC. There are still many unsolved problems in the development of a new strategy, including the absence of an integral picture of morphological changes in the tumor under the influence of mitochondrial therapy (MCT), and this determined the purpose of the study. Methods: The study included male and female Balb/c Nude mice weighing 21-22 g. B16/F10 melanoma was transplanted to mice subcutaneously, and suspension of live liver MC was injected intraperitoneally. MC were isolated by differential centrifugation (Avanti J-E high-speed centrifuge, BECKMAN COULTER, USA). After an MCT cycle, the melanoma structure was examined with hematoxylin and eosin staining and light microscopy (LEICA DM LS2). Results: Control samples of melanoma (without MCT) showed abundant vascularization with dense growth of epithelial-like cells with branched cytoplasm, an eccentric nucleus and a moderate content of melanin pigment. Massive hypoxic death, autolysis and autoxidation of tumor cells were registered in male mice after MCT, as well as matrix hyalinosis, pronounced subcutaneous tissue edema with segregation and necrotization of fat accumulations. The formation of a large cavity filled with detritus indicated signs of total tumor necrosis and ischemia. The accumulation of hyperchromic granules indicated the activation of granulocytic and phagocytic elements in the mechanism of tumor regression. Female mice showed rare small areas of necrosis, as well as pronounced fragmentation of nuclei and cytoplasm of cells in most fields of view induced by bioenergetic hypoxia, with the formation of apoptotic bodies. Along the edge of the cells subjected to autophagocytosis, melanin accumulated as it left the cells and accumulated as large granules, while small ones filled the space of the vessels. The process of melanoma regression was confirmed by a significant increase in free fibrous areas, where only faint cell shadows remained. Conclusions: The initial state of melanoma degrades under the influence of MCT inducing bioenergetic hypoxia in male and female Balb/c Nude mice in different ways. Gender, in fact, hormonal differences determine the dominance of different ways of cell death, necrosis (in males) and apoptosis (in females), while maintaining the MCT access to the processes of preventing tumor growth.

Aim. Our aim has been to develop an experimental model of the tumor growth against the background of hypothyroidism in rats of both genders in order to study possible influence made by hypothyroidism on progression of malignant tumors of various histological types. Materials and methods. In our studies we have used 100 outbred albino rats of both genders, with an individual body mass of 150-180 g. The female rats (n=30) and the male rats (n=30) have received Mercazolil at a day dosage of 2,5 mg/100g of the body weight for 30 days. After hypothyroidism in the treated rodents had been confirmed, one group of them (15 females and 15 males) were subcutaneously inoculated with the Guerin’s carcinoma cells, and another group (covering other 15 females and other 15 males) has been undergone to transplantation of the Sarcoma 45 cells. The reference group has included the rats of both genders with subcutaneously inoculated the Guerin’s carcinoma cells (n=10 females and n=10 males) and Sarcoma 45 cells (n=10 females and n=10 males), but without reproduction of the hypothyroidism model. Upon expiration of one month, within the 3 day period, we have estimated with a radioisotope analysis (RIA) standard assay kits (Immunotech, Czekh Republic) the levels of the thyroid hormones in blood of the tested animals as follows: Triiodothyronine (T3) (pM/L), total Thyroxine (T4) (pM/L) and Thyroid-Stimulating Hormon (TSH) (μU/mL). The obtained data have been processed with Statistica 10.0. Results. Upon the treatment with Mercazolil, we have found in the females a decrease by a factor of 7,3 in the total level of Thyroxine and an increase by a factor 1,6 in the TSH level (p<0,05), while in the males we have recorded a reduction by a factor of 2 in the total level of Thyroxine and an increase by a factor of 1,5 in the TSH level (p<0,05). In this case, the average sizes of the tumors in the female rats with Guerin’s carcinoma and hypothyroidism have been found smaller than those found in the reference group as given below: upon expiration of 4 days they are 1,3 times smaller (p<0,05), upon expiration of 7 and 10 days the volumes have been found 1,4 times smaller (p<0,05); upon expiration of 14 days the volumes have been recorded to be 1,5 times less (p<0,05); upon expiration of 18 days they have been reported to be 1,3 times less (p<0,05), and upon expiration of 21 days they have been estimated to be 1,4 times less (p<0,05). As to the males with Guerin’s carcinoma and hypothyroidism, the average sizes of their tumors as against the reference group data have been recorded to be smaller as follows: upon expiration of 4 days they are found 13,3 times less; upon expiration of 7 days they have been recorded to be 7,5 times smaller; upon expiration of 10 days the volumes have been estimated to be 1,9 times less (p<0,05), and upon expiration of 14 days they have been found to be 2,6 times less. The survival rate in the female rats in the main test has been recorded to be 1,6 times higher (p<0,05) against the data in the reference group, while the survival rate in the males has not shown any significant differences therein. In the female rates with S 45 growing against the background of hypothyroidism the average sizes of the tumors have been found to be less than those identified in the reference group as follows: after 4 days, the sizes have been recorded to be 1,4 times less (p<0,05); after 7 and 10 days they have been recorded to be 1,6 and 3,2 times smaller, respectively (p<0,05); after 14 days they have been found to be 3,9 times less, and after 18 days they have been recorded to be 4,8 times less. In the males at tumor growth week stage 1, the tumor sizes have increased 3,1 times as against 4 days of the tumor growth; upon expiration of 10 days the sizes have been found to be 7,1 times greater as compared with the previous period; upon expiration of 2 weeks they have increased 1,5 times (p<0,05); upon expiration of 18 and 21 days the tumor sizes have been recorded to be greater by a factor of 2,3 and by a factor of 1,6, respectively (p<0,05). The life spans in the female rodents in the main test group has been reported to be longer by a factor of 1,8 (p<0,05) than it has been the case with the reference group, and the average life span in the males has reached 21 days. Conclusion. We have revealed that in the female rates diagnosed with hypothyroidism the sizes of the subcutaneous tumor nodes of Guerin’s carcinoma and S 45 show slower progression as against that recorded in the reference group, and the life span recorded in the above rodents has been found as significantly longer, while in the male rats with hypothyroidism we have observed an irregular, slower, progression of the tumor nodes of Guerin’s carcinoma and S 45 within the period of 14 days, but subsequently we have detected the same progression rate as it is the case with the reference group data.

e21571 Background: Mitochondria of tumor cells undergo adaptive changes for even more active reproduction of tumor cells in the acidic and hypoxic microenvironment of tumor tissue. The purpose of this study was to evaluate antitumor effect of mitochondrial therapy in BALB/c Nude mice of both genders with B16/F10 melanoma growth. Methods: Male (n = 10) and female (n = 10) BALB/c Nude mice were injected subcutaneously with 0.5 ml of a suspension of B16/F10 mouse melanoma tumor cells in saline at a dilution of 1:20. Mitochondria (MC) were isolated from rat liver. 24 hours after subcutaneous transplantation of B16/F10 melanoma, mice were intraperitoneally injected with freshly isolated mitochondria (3.3 mg of protein per animal in 0.3 ml of saline). Further, mitochondrial therapy (MC therapy) was carried out according to the scheme on days 3, 5, 9, 13, 16, 19, 21. Male BALB/c Nude mice with subcutaneous inoculation of B16/F10 melanoma receiving intraperitoneal injections with 0.3 ml of saline served as controls. Results: Subcutaneous tumors in mice of both genders could be determined on the 5th day from the moment of tumor transplantation. In males, the regressive effect of MC therapy was recorded from the 8th day of tumor growth. The average tumor volume in males with MC therapy on day 8 was 3.0 times less than in the control group. On days 12 and 15, the regressive difference in tumor volumes between the groups was 1.8 times (p < 0.05) (MC therapy). On day 19, a slower tumor growth was recorded in the group with MC therapy, where the tumor volume was 2 times lower compared to the control volumes. At the end of the experiment on day 22, the difference in the average tumor volumes was 3.2 times, i.e. a significant inhibition of tumor growth was determined in males treated with MC therapy. A sizeable tumor node in females treated with MC therapy formed longer than in males, remaining as a flat tumor spot for up to 8 days. The inhibition of the growth of a large tumor node was determined on day 12 of melanoma growth, and the difference with the control values was 1.8 times (p < 0.05). On day 15, the tumor volumes in females with MC therapy decreased by 2.2 times compared with the control values, on day 19 - by 2.1 times. As a result, by the end of the experiment (day 22), the difference with the control group was 2.7 times. Conclusions: MC therapy inhibits the growth of B16/F10 melanoma in male and female BALB/c Nude mice. Formation of a sizeable tumor node and the start the tumor growth inhibition differs in time depending on the gender of experimental animals.

Topicality. Studies of the processes of the regeneration in the liver are not new, but however the development of the technologies of bio-therapy is capable of shifting the conventional approaches to the regeneration of the lost tissues. Mitochondrial therapy (MCT) occupies one of the highly important places in the list of bio-technologies by offering some unique mechanisms of the restoration of the organs. The liver having its specific tissue property of the regeneration demonstrates upon completion of hepatoectomy mitochondrial insufficiency and inhibition of the regenerative processes. Therefore it is required to evaluate the efficacy of mitochondrial stimulation in an experimental model in vivo with the proper morphological confirmation of the productivity and acceleration of the regenerative function of the liver that has become decisive for our research study work. Materials and methods. We have resected in 30 albino outbred male rats the left lobe of the liver, which is the maximum tolerable volume, amounting to 70% of the total mass of this organ, and upon of expiration of 24 hours we have transplanted to them intraperitoneally mitochondria harvested from a donor rat. Mitochondria have been isolated with the use of the differential centrifugation with high-speed refrigerated centrifuge Avanti J-E, BECMAN COULTER, USA. The morphological ORIGINAL RESEARCH examination of the liver sections has been conducted after their embedding in paraffin wax and staining with hematoxylin-eosin using microscopic testing of the liver sections with Axiovert (Carl 44 Zeiss, Germany) and applying the visualization software Axiovision 4 (Carl Zeiss, Germany). Results and discussion. It has been established that within the same period of time the quantitative data on the mass of the liver and its stump after MCT have exceeded the respective reference data by a factor of 1,6 (р˂0,05). The efficacy of the MCT application with respect to the regenerative function of the liver has been confirmed by the quality of the hepatic self-organization and replenishment of the key structures. A considerable supply of the most important cell elements of the connective tissue to the capsulated zone of the stump of the liver, an activation of neo-angiogenesis, hyperplasia and hypertrophy of hepatocytes with the repetitive order of the arrangement of the lobe structure of full value are manifestations of the acceleration of the regeneration. Conclusion. The high energy potential of fresh mitochondria makes possible to realize the trigger mechanisms of the bio-synthetic processes addressing an immediate restoration of the damaged or disordered liver tissues that can hold the greatest promise in emergency surgery and the activation of slowly progressive processes.