Analysis of the Link between rs4977574 Single Nucleotide Polymorphism of the Long Non-Coding RNA ANRIL Gene and Prostate Cancer Development

The purpose of the study was to investigate the possible association between ANRIL gene rs4977574-polymorphism and prostate cancer occurrence among men of the Ukrainian population. Materials and methods. A total of 250 males were enrolled in the study. Of these, the experimental group included 184 prostate cancer patients, and the control group included 66 men without a history of malignant tumors. Genotyping of the ANRIL rs4977574 locus was performed by real-time polymerase chain reaction. The reaction was performed on a Quant Studio 5 DX Real-Time instrument (Applied Biosystems, USA) in the presence of TaqMan assays (TaqMan®SNP Assay C_31720978_30). The genotyping results were statistically processed using the SPSS software package (version 17.0). Values of p less than 0.05 were considered as statistically significant. Results and discussion. ANRIL (Antisense Non-coding RNA in the INK4 Locus), also known as CDKN2B-AS1, is a long non-coding RNA (3.8-kb) transcribed from the short arm of the human chromosome 9 (p21.3). ANRIL transcripts promote their main molecular effects through interaction with proteins of Polycomb repressive complex 1 and Polycomb repressive complex 2. Ultimately, this leads to epigenetic cis-inactivation of the tumor growth suppressor genes located in the Chr9p21 region: CDKN2A/p16INK4A, CDKN2A/p14ARF, CDKN2B/p15INK4B. Recent experimental studies have demonstrated the involvement of ANRIL in the development of malignant tumors of different localization. At the same time, there is almost no information about the role of the gene polymorphisms of this RNA in the occurrence of prostate cancer. The possible link between ANRIL gene polymorphism and prostate cancer risk in the Ukrainian population is not fully understood. It was found that the control men and prostate cancer patients did not differ significantly in the frequency of rs4977574-genotypes (p = 0.886). No significant difference was found during the corresponding comparison separately among persons with normal weight, overweight, without, and with the habit of smoking (p >0.05). Analysis of the association of different rs4977574 genotypes of the ANRIL gene with the risk of prostate cancer using logistic regression also did not show a reliable relationship under different models of inheritance, both before and after adjustment for age, body mass index and smoking (p >0.05). Conclusion. Thus, for the first time, we performed an analysis of the relation between ANRIL gene polymorphism and the development of malignant tumors of the genitourinary system in the Ukrainian population. The results showed that the polymorphic locus rs4977574 is not associated with the risk of prostate cancer

Features of Blood Supply to the Bones of the Lower Leg in the Ankle Joint Region

The purpose of the study was to investigate the topography of permeable (nutrition) arteries in the lower third of leg during human ontogenesis. Materials and methods. The research was conducted on 30 specimens of human prefetuses and fetuses aged 3-9 months of prenatal development; 28 bone specimens of adult tibia and fibula and 27 series of computerized tomography scans of lower extremities of adults aged 21-72 were used. We have used a complex of methods for morphological research: anthropometry, morphometry, macromicroscopy, injections of blood vessels of X-ray contrasting mixtures, three-dimensional reconstruction and statistical analysis. Results and discussion. Cartilage osteogenesis occurs in the prefetal period of prenatal human development, in which blood vessels play a crucial morphofunctional role. During endochondral ossification of the tibia, the vessels of the bone cuff grow into the diaphysis of the cartilaginous model of the bone and osteogenic cells go beyond their limits. In the third trimester of prenatal development, blood vessels grow into the epiphyseal part of the cartilaginous model and an epiphyseal centre of ossification forms. Between the epiphysis and diaphysis centers of ossification, a metaphysical growth plate forms that develops intraosseous anastomosis between the diaphyseal and metaphysical blood vessels. In the metaphysis region, there are extra-skeletal anastomoses. Often, the insertion of nutrition arteries into the bone of the tibia is usually the middle third of bones (55%) and distal metaphysis (37%). In 10%, there were nutrition foramina in the upper third of the tibia. Three-dimensional reconstruction also clearly shows a more extensive and developed network of vessels in the bone marrow canal in metaphysical regions. In the middle third of the tibia and fibula, 1-3 nutrient foramina can be traced on most specimens. On the fibular, in addition to the posterior-medial localization of the nutrient arteries, there were variants with a "high" (above the metaphysis) placement of the entrance of the vessel into the bone substance. Conclusion. The most common places where arteries enter the bones of the lower leg are the middle third of the bones (55%) and the distal metaphysis (37%). In 10%, there were nutrient foramina in the upper third of the tibia. The three-dimensional reconstruction also clearly shows a more extensive and developed network of vessels in the bone marrow canal in metaphysical regions. During the postnatal period, we most often encountered the localization of nutritional arteries on the anterior-lateral surface of the tibia (23%), or their combination – the presence of both nutrient arteries on the posterior and medial bone surfaces (18.3%)

Influence of Low Concentrations of Carbon Monoxide on Metabolism of Isolated Heart under Conditions of Ischemia-Reperfusion

The purpose of the study was to determine the effect of different concentrations of carbon monoxide on the metabolism of isolated mice hearts. Materials and methods. To elucidate the effect of low concentrations of carbon monoxide on the myocardium, we performed retrograde perfusion of isolated hearts of laboratory mice with Krebs-Henseleit solution, which was saturated with carbon monoxide for 5, 10, and 30 minutes. We then determined how different concentrations of carbon monoxide affected coronary volumetric flow rate, myocardial glucose and calcium uptake, creatinine release, and aspartate aminotransferase release. During perfusion, R-wave amplitude and R-R interval were measured using an electrocardiograph. To determine the effect of ischemia on the heart muscle during perfusion with solutions of different concentrations, we measured the area of the affected myocardium after staining with 2,3,5-triphenyltetrazolium chloride. Results and discussion. After these studies, it was found that different concentrations of carbon monoxide had a dose-dependent effect on the isolated mouse heart. However, the dependence of the effects does not follow the pattern «lowest concentration – lowest effect». At the same time, an increase in concentration did not mean an increase in adverse effects on the myocardium. Even on the contrary, the smallest concentration led to increased signs of ischemic myocardial damage. In particular, the use of the solution, through which carbon monoxide was passed for 5 minutes, caused vasoconstrictor effect during perfusion. At the end of reperfusion, vasoconstrictor effect was observed after using a solution through which carbon monoxide was passed for 10 minutes. Increased glucose uptake was observed in the group with 30-minute carbon monoxide permeation against the background of the minimal myocardial creatinine release. In this group there was also a decrease in Ca2+ loss at the beginning of reperfusion (immediately after ischemia). The above phenomenon explains the least degree of ischemic myocardial damage in the isolated mouse heart. The obtained data should be expanded. Since it is difficult to accurately determine the dose of carbon monoxide, then the use of donor compounds is promising. Such compounds include CORM-2 and CORM-3. Under physiological conditions, they decompose in a controlled manner, releasing a specific amount of carbon monoxide. Conclusion. The obtained results indicate that at different concentrations of carbon monoxide can differently influence different structures of cardiomyocyte: at one concentration it binds to calcium channels, other concentrations influence ion channels of plasma membrane, which can explain all these dependencies

Study of Correction of Somatic Pain under the Conditions of Experimental Pathology of Multiple Sclerosis

The purpose of the study was to experimentally substantiate the ways of pharmacological correction of somatic pain syndrome in conditions of the experimental equivalent of multiple sclerosis through a comparative system analysis and the use of complex methodological approaches. Materials and methods. To study multiple sclerosis, we used an experimental model with autoimmune mechanisms of inflammatory demyelination – a model of experimental allergic encephalomyelitis. To assess the antinociceptive activity of painkillers, we used the method of electrical stimulation of the rats’ tail root. The activity of the enzyme prostaglandin H-synthetase was also determined. Results and discussion. A comparative analysis of the analgesic activity indicators of combinations of methylprednisolone with analgesics under the condition of the formed experimental allergic encephalomyelitis showed that their antinociceptive potential (taking into account the basic therapy with methylprednisolone) decreased in the series meloxicam > lornoxicam ≈ ketorolac ≈ paracetamol > celecofenacoxib ≈ sodium diclofupene ≈ diclofupene ≈ diclofupene. Accordingly, the maximum effect on the threshold of nociception under these experimental conditions was exerted by meloxicam and lornoxicam. The combined administration of methylprednisolone with diclofenac sodium, celecoxib and meloxicam reduced the activity of prostaglandin N-synthetase in the brain structures by 49.8% (p <0.05), 50.4% (p <0.05) and 51% (p <0.05), respectively, compared with the indicators of the control group. The same drugs markedly reduced the activity of prostaglandin N-synthetase in the spinal cord by 23.9% (p <0.05) (Methylprednisolone + diclofenac), by 34% (p <0.05) (Methylprednisolone + celecoxib) and by 47.4% (p <0.05) (Methylprednisolone + meloxicam) compared with the control group. Our analysis of the analgesic activity of antidepressants and anticonvulsants as means of correcting nociceptive pain in experimental allergic encephalomyelitis found that their antinociceptive potential was inferior to the severity of the analgesic effect of nonsteroidal anti-inflammatory drugs. Conclusion. Among the studied non-steroidal anti-inflammatory drugs, antidepressants and anticonvulsants, the maximum therapeutic efficacy as a means of correcting nociceptive pain in experimental allergic encephalomyelitis against the background of basic methylprednisolone therapy was shown by meloxicam, which gives grounds to recommend it as the analgesic of choice for eliminating somatic pain syndromes

Changes in Glycanic Determinants of Lymphocytes Membranes in Peripheral Blood in Patients with B-Cell Chronic Lymphocytic Leukemia under Antitumor Therapy

The purpose of the study was to study the nature of changes in the exposure of surface glycans of peripheral blood lymphocytes in patients with B-cell chronic lymphocytic leukemia under conditions of antitumor therapy. Materials and methods. We studied the features of exposure of surface glycotopes of peripheral blood lymphocytes in patients with B-cell chronic lymphocytic leukemia under conditions of antitumor therapy using a set of seven lectins labeled with FITC and monoclonal antibodies to Tn-antigen- FITC for the detection of Tn antigen and CD43 exposure on blood lymphocytes. Cytostatic therapy included cyclophosphamide, vincristine (oncovin), prednisolone. Data were recorded on a Beckman Coulter EPICS flow cytometer. The results were processed using FCS3 Express. Results and discussion. The number of lymphocytes of healthy donors with a positive reaction to ConA, PHA-L, SNA, MAA-II and α1-acid glycoprotein amounted to 16.0±3.0%, 23.0±2.3%, 15.0±1.5%, 25.0±1.8% and 15.0±1.3%, respectively. The number of LABA-, UEA I-positive lymphocytes was 0.90±0.03% and 2.9±0.2%, respectively, and there was no binding to antibodies to Tn- and CD43-antigens. In the blood of patients with chronic lymphocytic leukemia, the level of ConA-, SNA- and MAA-II-positive lymphocytes increased relative to control by 2.2, 3.7 and 2.6 times, respectively. The number of LABA- and UEA I-positive lymphocytes in patients with chronic lymphocytic leukemia increased by 11 (p <0.01) and 23 (p <0.001) times and amounted to 10.5±0.5% and 67.5±5.5% respectively. The number of lymphocytes with CD43 antigen on their surface increased by 72 times, and the Tn antigen increased by 80 times. Cytostatic therapy reduced the level of LABA- and UEA I-positive lymphocytes by almost half, and MAA II-positive cells and lymphocytes interacting with antibodies to CD43 and Tn antigen by a third. The level of PHA-L-positive lymphocytes in the blood of chronic lymphocytic leukemia patients after undergoing alkylating therapy increased by 18.0±2.0% and almost did not differ from those obtained in the control group. Conclusion. 1. In chronic lymphocytic leukemia patients, the structure of glycoconjugates in peripheral blood lymphocytes changes, manifested in increased exposure of L-fucose, α-mannose and N-acetylneuraminic acid, which is confirmed by a significant increase in relation to the control of the number of ConA-, SNA-, MAA-II-, LABA I-positive cells. 2. Patients with chronic lymphocytic leukemia showed a significant increase in the number of lymphocytes, in which the markers of carcinogenesis CD43 and Tn antigens were found. 3. Cytostatic therapy significantly reduced the level of LABA-, UEA I- and MAA II-positive cells, as well as partially Tn- and CD43-antigen-positive lymphocytes, which indicates its positive effect on the treatment of chronic lymphocytic leukemia

Dynamics of Changes in the Indicators of Mathematical Analysis of Heart Rate in Conditions of Adrenaline Myocardial Dystrophy against the Background of Different Functional Activity of the Pineal Gland

The purpose of the study was to examine the features of heart rate variability in experimental hyper- and hypofunction of the pineal gland, complicated by adrenal myocardial dystrophy. Materials and methods. To solve the set tasks, three models of functional states were created: hypo-, hyperfunction of the pineal gland, adrenaline myocardial dystrophy. Hypermelatoninemia was reproduced by administering melatonin. The hypofunction of the pineal gland was simulated by round-the-clock lighting. An experimental model of cardiac pathology, namely adrenaline myocardial dystrophy, was reproduced by administering adrenaline hydrochloride. The degree of tension of regulatory mechanisms and mechanisms of nervous regulation was assessed by mathematical analysis of heart rate variability. Results and discussion. The task of mathematical analysis is to extract “hidden information” and thereby assess the state and degree of tension of the regulatory mechanisms of the whole organism. According to the authors, mathematical analysis allows a differentiated approach to the assessment of regulatory effects, which is done by the nervous autonomic system, subcortical structures and the cerebral cortex. The data obtained by the authors suggest that the introduction of adrenaline in conditions of hypofunction of the pineal gland have cause-and-effect changes. These changes lead to disorders of autonomic regulation and possibly a trigger mechanism for a decrease in melatonin production by the pineal gland. Studies have shown that one of the advantages in conditions of hyperfunction of the pineal gland is the dominant role of parasympathetic processes in the mechanisms of adaptation of the heart to conditions of adrenal myocardial dystrophy. Timely and sufficient activation of melatonin provides a high efficiency of compensatory-adaptive changes and a positive role in increasing myocardial resistance. Conclusion. The results of the study showed that the dynamics of changes in the values of the indicators of the mathematical analysis of the heart rate in adrenal myocardial dystrophy against the background of hypo- and hyperfunction of the pineal gland revealed unidirectional changes. These changes are manifested in a decrease in heart rate, stress index and vegetative balance indicator. Consequently, the results obtained suggest that the introduction of melatonin against the background of adrenal myocardial dystrophy has a general corrective effect on the restoration of the functional state of the heart. Based on the results, the authors consider it appropriate to continue the study of the influence of various stressors on the functional state of the heart in conditions of different physiological activity of the pineal gland

Changes of Rat Embryon Nephrogenesis during Cadmium Salt Intoxication in Pregnant Females

The purpose of the experimental study was to determine morphogenetic disorders of embryonic and fetal kidney development in chronic intragastric exposure to cadmium salts (cadmium chloride, cadmium citrate) in pregnant females. Materials and methods. Low doses of cadmium salts were selected for the study, which can be compared with the actual concentration of cadmium in the daily diets of women, including pregnant women, in industrial regions. In the experiment, female rats with a given gestational age were divided into groups as follows: Group 1 – control (number of females – n = 16, of which 8 left the experiment on the 13th day of pregnancy, and 8 on the 20th; the number of embryos – n13 = 76; n20 = 77); Group 2 – administration of cadmium chloride at a dose of 1.0 mg/kg body weight of the female (number of females – n = 16; number of embryos – n13 = 65; n20 = 62); Group 3 – administration of cadmium citrate at a dose of 1.0 mg/kg body weight of the female (number of females – n = 16; number of embryos – n13 = 69; n20 = 70). Results and discussion. New quantitative data on the effect of cadmium salts were obtained on the thickness of the cortical and cerebral layers of the kidneys, the diameter and area of the cavity of the nephron capsule in the prenatal period of development in chronic female intoxication. On the 13th day, the effect of cadmium salts on the development of embryonic kidneys was multidirectional: the effect of cadmium chloride led to an increase in the thickness of the mesonephros and mesonephric duct, and the effect of cadmium citrate reduced the studied parameters. On the 20th day of rat embryogenesis in the group exposed to cadmium chloride, the renal weights increased, and when exposed to cadmium citrate, the weight of the kidneys decreased significantly (p˂0.05) both in comparison with control values and in the group exposed to cadmium chloride. On the 20th day of development, in order to exclude an error in estimating the dynamics of changes in the weight of embryo mass and kidney mass, the nephrofetal index was calculated, i.e. the ratio of wet kidney mass to wet weight of fixed fetus. Histological parameters of nephron diameters were also compared. Conclusion. Cadmium citrate has been shown for the first time to be less non-photoxic than cadmium chloride. Calculation of the area of the cavity of the nephron capsule by the spline contour method showed a 2.6-fold decrease in the average area of the capsule cavity in the group exposed to cadmium chloride relative to the control mean values, indicating a violation of nephrogenesis. When exposed to cadmium citrate, there was also a decrease in the area of the cavity of the nephron capsule by 1.8 times, which indicates a lower level of nephrotoxic cadmium citrate compared to cadmium chloride, despite the identity of the dose of exposure

Change of Embryotoxic Effects of Metal Citrates Depending on the Duration of Their Introduction

Given the increase in cadmium in the environment (air, soil, water) through modern industrial processes, the absorption of significant amounts from cigarette smoke is relevant to studying the effect of cadmium compounds on embryogenesis. The purpose of the study: experimental study and comparison of embryolethality and embryotoxicity of metal citrates depending on the duration of their intragastric administration (during 13 and 20 days of gestation). Materials and methods. The study was performed on 120 white adult female Wistar rats that weigh 170-200 g. They were divided into 6 groups due to the intragastric administration of solutions of the studied metals – rats treated with citrates: cadmium at a dose of 1.0 mg/kg – 1st group (nfemale = 20, nemb = 166); cadmium at a dose of 1.0 mg/kg and cerium (1.3 mg/kg) – 2nd group (nfemale = 20, nemb = 185); cadmium in a dose of 1.0 mg/kg and germanium (0.1 mg/kg) – 3rd group (nfemale = 20, nemb = 184); cadmium at a dose of 1.0 mg/kg and zinc (1.5 mg/kg) – 4th group (nfemale = 20, nemb = 179); cadmium at a dose of 1.0 mg/kg and nanocomposite (iodine + sulfur + selenium) at a dose of 2.0 mg/kg – 5th group (nfemale = 20, nemb = 180), 6th group – control (nfemale = 20, nemb = 212) – proportional volume of sterile saline in the same way. Females were divided into 2 subgroups of 10 animals each, depending on the duration of administration of test substances. The embryotoxic and embryolethal effects of the test substances were evaluated according to generally accepted criteria, which were calculated according to well-known formulas. Results and discussion. Cadmium compounds harm the embryogenesis of rats in the experimental groups and increase the rates of embryolethality. The most pronounced differences in these parameters concerning the control were found in animals with isolated exposure to cadmium citrate: the rates of total embryonic mortality increased by 4.0 times in both study periods of embryogenesis and 20% from the 13th to the 20th day; increased pre-implantation mortality by 6.0 times with an increase in post-implantation mortality by 3 times on the 13th day and by 15.0 times and 2.8 times on the 20th day of embryogenesis, respectively. Depending on the duration of introduction, the indicators of pre-implantation mortality increased by 25.0% with an increase in post-implantation mortality by 22.2% in the same period. The experimental group of exposure to cadmium citrate at a dose of 1.0 mg/kg recorded the lowest indicators of the number of live fetuses and intrauterine survival with the highest resorption rate studied of embryonic development. At the same time, in the groups of combined exposure to cadmium citrate with metal citrates, a decrease in the indicators of total embryonic mortality was 50.0% - 30.0%, pre-implantation mortality – 50.0% - 25.0%, post-implantation mortality – 60.0% - 44.4 % and increase in the number of fetuses per female – 12.7% - 25.3%. Conclusion. Analysis of the results shows a pronounced embryotoxic effect of cadmium citrate at a dose of 1.0 mg/kg on the processes of embryogenesis, which is a significant increase in overall embryonic mortality, preimplantation, and postimplantation mortality compared with the control group in all studied terms. In the groups of combined action of cadmium citrate with metal citrates, the data obtained indicate a decrease in the accumulation of cadmium under the influence of the studied citrates, which allows them to be considered as potential bioantagonists of cadmium citrate

Dynamics of Power Indicators Growth in Bodybuilders of 18-19 Years Old at the Stage of Specialized Basic Training

The purpose of the study was the analysis of the growth dynamics of power indicators of athletes-bodybuilders at the stage of specialized basic training. Materials and methods. The study was conducted from March to the end of June (4 months) 2021. It involved 60 athletes aged from 18 to 19 years. Three research groups were formed, 20 athletes in each. These groups of athletes were divided by the usual sample method and by age. Qualification and anthropometric characteristics did not differ significantly. The only difference was in the proposed training programs. Theoretical analysis of scientific and methodological literature was carried out. Test control method for determining the maximum muscle strength of athletes to assess the initial level of muscle strength development in the surveyed contingent and determine the characteristics of its dynamics, was used. The obtained data were used to calculate the indicators of the value of the training load of athletes. In addition, method for quantitative assessment of athletes' physical load was used. The mathematical processing of research results was carried out using the software packages IBM SPSS Statistics 20. Results and discussion. The research results indicate that the values of the training loads indicators of bodybuilders and the nature of their changes in conditions of the same level of athletes' fitness and the structure of the training session depend on the characteristics of the selected training means (training programs). The analysis of the results obtained shows that under the conditions of the second variant of the training program, the indicators of the working mass of the athletes' equipment change more significantly during the entire research period. A similar tendency is observed when monitoring the load volume indicators, despite the fact that the most significant increase in this indicator among bodybuilders when performing formative exercises was obtained under the conditions of using the third variant of the training program, using the “premature fatigue” method. Conclusion. The growth rates of the strength capabilities of the main muscle groups turned out to be the highest among bodybuilders of the second main group due to the long-term use of the “premature fatigue” method. When performing exercises of a formative nature, the growth rate of strength capabilities was 30.9% (p <0.05), while when performing exercises of a basic nature, such an increase was three times less (by 10.5%, p <0.05)

Reactivity of the Blood Microcirculation System in Primary School Children to the Temperature Factor

The purpose of the study was to investigate the adaptive capacity of the blood microcirculation system in children of primary school age for a functional test with heating. The article deals with the study of the blood microcirculation system in primary school children by laser Doppler flowmetry and the study of the reserve capacity of tissue blood flow in children on a test with increasing temperature factor. Materials and methods. The research was conducted on the basis of the laboratory of physiological research of the Department of Anatomy and Physiology of People and Animals of Bohdan Khmelnytsky Melitopol State Pedagogical University. The surveyed contingent consisted of 80 practically healthy children aged 6 to 9 years old, secondary school students in Melitopol. Based on a comprehensive morphofunctional study, we obtained data on individual-typological features of blood microcirculation in primary school children and the reserve capacity of tissue blood flow using functional tests. It is established that the adaptive features of the blood microcirculation system in boys and girls of early school age are due to different levels of local and central mechanisms of microvessel regulation. Results and discussion. The results obtained during the thermal test indicate different levels of reactivity of the body to increasing the temperature factor and the neurogenic vasoconstrictor effects of the sympathetic nervous system on the vessels of the arteriolar part of the microcirculatory tract. The reaction to the thermal test directly depends on the individual-typological features of blood microcirculation. Conclusion. At a ratio of reaction to thermal influence in children with various types of microcirculation, the maximum increase in tissue perfusion was observed in children with type III of laser Doppler flowgram. This level of microvascular reactivity, as well as the relatively rapid recovery of blood flow after thermal hyperemia is explained by the fact that children with hyperemic type of microcirculation have increased microvascular tone due to neurogenic effects of the sympathetic chain of tissue blood flow regulation. Children with the hypoemic type of microcirculation, on the other hand, have a somewhat reduced sympathetic tone relative to the normoemic type, as a result of which the reserve of capillary blood flow is reduced and the time of blood flow recovery after thermal exposure is prolonged