Visnyk of Dnipropetrovsk University. Biology, medicine

8-12

Authors: T. V. Martyshuk

This article presents the results of research on the influence of oxidative stress on the intensity of the process of lipid peroxidation and the activity of the glutathione system of antioxidant defense in the organisms of rats. Intramuscular injection of 50% solution of tetrachloromethane at a dose of 0.25 ml per100 gof body weight to rats from the experimental group causes activation of the process of free radical lipid oxidation with excessive accumulation of intermediate anf final products of lipid peroxidation. The research results indicate that the development of oxidative stress leads to significant and probable acceleration of the formation and accumulation in the plasma of the rats, in all stages of the experiment, of lipid hydroperoxides and malonic dialdehyde. The highest level of hydroperoxides of lipids in the blood plasma of rats under oxidative stress was on the second day of the experiment, where it was 843 unE/ml, whereas in the control this index was 0.245 unE/ml. During the research into the content of malondialdehyde it was found that in the experimental group of animals it was 2.03 times higher than in the control group on the 5th day. On the 10th and 14th days of the experiment we observed a slight reduction in the levels of lipid hydroperoxides and malondialdehyde. The development of oxidative stress also leads to inhibition of the glutathione system of antioxidant defense in the rats’ organism. This shows the low activity of glutathione peroxidase and the low level of restored glutathione in the blood of the rats from the experimental group. On the 5th day of experiment the activity of glutathione peroxidase and restored glutathione level in the blood of the rats which were injected with carbon tetrachloride was at its lowest, compared with the control these indices decreased respectively by 53% and 51%. On the 10th and 14th days of the experiment the activity of glutathione peroxidase and restored glutathione level in the blood of the rats from the experimental group were slightly increased, but compared to the control they were still significantly lower. Significant disturbances were found in the oxidation-antioxidant balance of the animals under oxidative stress, which is characterized by the activation of the processes of free radical lipid oxidation with excessive accumulation of intermediate and final products and the inhibition of the antioxidant defense system.

18-26

Authors: T. O. Tretiak, O. V. Severynovska, M. Boyko

The contents of education must correspond not only to the level of knowledge and competencies, but also be directed to self-determination of the personality and creation of conditions for its self-development and self-realization. The aim of this study was to identify the psycho-physiological peculiarities of Biology students and to establish the connection between the students’ academic performance, their cognitive abilities and their psycho-physiological characteristics, which is essential for improving the academic process and bringing it to the high level of professional education necessary for graduates of higher educational establishments to compete successfully in the job market. It was established that the majority (46%) of Biology students are ambiverts, 34% of them are extroverts, and 18% – introverts. There is a statistically reliable connection between the academic grades and the personality type: the more successful students are ambiverts and introverts. Extroverts demonstrate lower academic results. According to different types of asymmetry manifestation among Biology students, the right-handed accounted for 35% of the tested, those with prevailing right hand – 51%, those with prevailing left hand – 1.2% and the left-handed – 12%. There is no dependence of academic performance on the asymmetry manifestation. Most Biology students have a sanguine-choleric temperament and slightly expressed or absent asymmetry, which testifies to the power of nervous processes and is a precondition for successful studying. The majority of students, irrespective of the academic performance level, have a low level of stress resistance and a medium level of personality anxiety. Students who score A and B points for academic performance, have a medium level of situational anxiety. Those who score C points have a high level of anxiety. There is no connection between the academic points and personality anxiety level, but there is significant connection between academic performance and situational anxiety. There has been established a significant dependence of academic performance on the level of situational anxiety: introverts and extroverts have a high level, ambiverts have a medium level of situational anxiety. Introverts and ambiverts with a low and medium anxiety level demonstrate higher academic performance. Extroverts with a high anxiety level find it more difficult to study. The majority of Biology students have a high level of visual-imaginative, object-active and creative thinking types. 83% of students have a high and a medium level of verbal-logical thinking type. Abstract-symbolic and intuitive thinking are developed in lesser degree. There has been established a reliable negative correlative connection between object-active thinking type and introversion, and a positive connection between intuitive thinking and introversion, irrespective of academic results. The more introversion is expressed, the more intuition is developed. Academically successful students demonstrated a positive correlation between the abstract-symbolic thinking type and introversion. Extroverts with low academic performance don’t have significant correlation connections with the thinking types, whereas it is characteristic for academically successful students to demonstrate a higher level of visual-imaginative, verbal-logical, abstract-symbolic and creative thinking along with an increase in demonstrating extroversion. Practically all thinking types (except for the object-active) are more fully expressed in successful ambiverts; the manifestation of a certain thinking type increases while ambiversion increases. Less academically successful students have a reliable correlative connection between ambiversion and visual-imaginative, abstract-symbolic and creative thinking. Performance of tasks connected with all thinking types irrespective of typological characteristics of students was accompanied by a reliable increase in saturation. More diversified reactions in the process of thinking activity were observed in the cardio-vascular system: introverts and ambiverts who demonstrated high results in the tests, showed significant decrease of heart beat rate in the process of intuitive and visual-imaginative thinking. Extroverts who demonstrated low results in the tests, by contrast, showed an increase in heart beat rate. When students of all typological groups were doing tasks connected with verbal-logical thinking, their heart beat rate significantly increased. Ambiverts and introverts were more successful. Thus, the whole range of psychological and physiological characteristics of a person defines his successful academic performance and real activity.

32-36

Authors: G. A. Al-Maali, N. A. Bisko, A. M. Ostapchuk

The aim of our research was to study the influence of citrate and sulfate of copper, manganese and zinc on the carbohydrates composition of biomass of the medicinal mushroom Trametes versicolor (L.) Lloyd. cultivated in a liquid medium. The studied strain of Trametes versicolor 353 was obtained from the Culture Collection of Mushrooms (IBK) from the M. G. Kholodny Institute of Botany, National Academy of Sciences of Ukraine. Citrates of copper, manganese and zinc were obtained from the Institute of Nanobiotechnologies and Resource Conservation of Ukraine, Kyiv. In this study we used glucose-peptone-yeast extract medium (GPY) with the following composition of (g/L): glucose – 25, peptone – 3, yeast extract – 3, K2HPO4 – 1, KH2PO4 – 1, MgSO4 ∙ 7H2O – 0.25, distilled water – 1,000 ml; pH 6,5 (control medium). Cu2+ (sulfate form or citrate form) were added to control medium in concentration 4 mg/L, Zn2+ and Mn2+ (sulfate form or citrate form) were added to control medium in concentration 1 mg/L. Mycelium was grown in a submerged culture on a rotary shaker (120 rpm) at 26 °C in 250 ml Erlenmeyer flasks, containing 50 ml of liquid medium. The biomass was harvested after 9 days of cultivation in the liquid medium. Monosaccharides and sugar alcohols was analyzed by high-performance liquid chromatography Agilent 1200 (Agilent technologies, USA). The results of our research demonstrated that the fraction of total carbohydrates from T. versicolor 353, which were cultivated on the control medium, consists of glucose (91.5%), galactose (1.9%), mannose (5.4%), fucose (0.6%) and xylose (0.7%). The adding of sulfate of zinc or copper to the liquid medium didn’t change the content of monosaccharides of T. versicolor 353 biomass relative to the control medium. At the same time, in the medium with citrate of zinc the amount of glucose increased and the amount of mannose decreased relative to the control medium. Xylose and fucose were not detected in this case. In mycelium of T. versicolor 353 that was cultivated in the medium with manganese sulfate or manganese citrate we detected a small amount of ribose. But the content of other monosaccharides didn’t change in the medium with sulfate of manganese. At the same time, the addition of citrate of manganese to the medium raised the amount of mannose and reduced the amount of glucose in the biomass of T. versicolor 353 relative to the control medium. Also copper citrate reduced the content of glucose in mycelium of T. versicolor 353 relative to the control medium. But in this case we detected mannitol and sorbitol in the biomass. This data indicates that synthesis of sugur alcohols in T. versicolor 353 depend on copper ions in bioactive forms of citrate.

43–47

Authors: E. Z. Ivanchenko

In studies on healthy humans we examined changes in the H reflex amplitude and the intensity of background EMG of the soleus muscle within the premotor period of voluntary extension of the contralateral ankle joint. The tested persons were in the lying prone position with relaxed legs, or in the standing position with support on both feet or only on one foot with additional hand support. The soleus H reflex was induced using the standard technique by transcutaneous stimulation (1 ms long current pulses) of the tibial nerve afferents in the region of the popliteal dimple. Using superficial electrodes, we recorded the integral EMG reflex discharge from the m. soleus, amplified, and visualized it using a two-channel digital oscillograph, Handiscope HS3 (TiePieEngeneering, Netherlands). In other tests, we also recorded tonic background EMG activity from the soleus muscle. After fullwave rectification and low-frequency filtration we estimated the area under the integral curve enveloping oscillations of the above mentioned EMG. In the lying position the extension of contralateral ankle joint evoked anticipatory facilitation of the testing soleus H-reflex. It manifested in 90–60 ms and reached the maximum 30 ms before the conditioning movement. Considerable changes of the soleus background EMG-activity did not occur during the premotor period. Consequently, it is supposed that the mechanism leading to the H-reflex facilitation is the decrease of presynaptic inhibition at 1a. In the standing position with support on both feet contralateral ankle joint extension caused anticipatory inhibition of the soleus H-reflex and simultaneous suppression of background EMG 90–60 ms before the conditioning movement. These results indicate the anticipatory changes of muscle tone and suggest participation of postsynaptic mechanism in the inhibition of the testing H-reflex. When the moving contralateral extremity had no support, the contralateral ancle joint extension did not cause anticipatory inhibition but facilitation of the soleus H-reflex. Its time course was similar to that observed in the lying position. The considerable changes of the soleus background EMG-activity did not arise. The results of the experiments indicate that the nature of premotor changes in the spinal neuron circuits, associated with the lower limb muscles, depends not only on the type of upcoming voluntary movement. Being caused by the contralateral limb movements, they depend greatly on the fulfillment or nonfulfillment of the support function by the lower limbs.

53–58

Authors: V. P. Zhalko-Tytarenko, E. A. Synetar, S. I. Savoschenko

It is well known that the ureter of mammals and man works as a flow system that normally remains uninfected. In biotechnology flow systems are used to produce microbial mass on orders greater than when grown in laboratory and industrial periodic cultures. Therefore, these systems are used in the microbiological industry. But there ina flow systems one can observe a growth in microbial populations, the patterns of which can be fruitful in studying the process of catheter-associated urinary tract infections. The main factors are the speed of propagation and speed of flow, and their variation in conditions of the catheterized urinary tract, which determines the magnification factor of microbial infection. In this lies the general biological need to study them from a biotechnology perspective. Hence the question arises – why are there changes in the flow system of the urinary tract during catheterization, which contribute to the reproduction of opportunistic microorganisms, as a factor of infection? In the study of the biological properties of agents of catheter-associated infections this issue has not attracted sufficient attention from researchers. In view of the above, the purpose of our study was to establish patterns of population growth of agents of urinary tract infections on the model of the yeast fungus species Candida albicans (C.P. Robin) Berkhout, 1923 in the catheterized human urinary tract, treated as a flow system. The results of the research show that the emergence of catheter-associated urinary tract infections is a regularity inherent throughout the flow system. It was proved that with the model C. albicans, bacteria in urine multiply faster than in the medium Saburo. The study established that one of the factors leading to catheter-associated urinary tract infections is the persistent presence of urine deposits around the head of the catheter in the bladder, which provides bacteria with sufficient conditions for reproduction and where bacteria cannot be eliminated by urine flow. When the pathogen enters and reproduces in the bladder, the latter is a reservoir of infection like a "royal culture", with a gradual overgrowth in the catheter of yeast fungi of the species C. albicans. These data suggest that the mechanism of strengthening catheter-associated urinary tract infections are the formation of biofilms by microorganisms around the catheter head and on its surfaces. It is recommended to further improve the design of urinal catheters in such a way that ensures the complete emptying of the bladder and avoidance of urethral reflux.

65–70

Authors: A. G. Morenko

An important component of the general biological problem of coordinated activity of the brain’s motor systems and of the executive apparatus during the implementation of motor functions is the question of the interconnection of electroencephalogram (EEG) rhythm characteristics, above all α-rhythm, with control peculiarities of manipulative movements (MM). A test group consisting of 113 healthy right-handed women from the ages of 19 to 21 was divided into two groups according to the average magnitude of their individual modal α-frequency – groups with high and low values of individual modal α-frequency (IαF). The ideal time of a simple sensorimotor reaction and choice-point behavior as well as speed capabilities of nervous processes during the finger tapping test, and measures of the power spectrum of ЕЕG spectral components individually determined for each testee in quiescent intervals and while performing alternating movements by the fingers of the right hand were evaluated. Alternate female finger movements were accompanied by a decrease in the EEG α2- and α3-activities in the posterior cortical areas, and β1- and γ-activities in the frontal, temporal and central areas of the cortex compared with the same in the immobile state; they were also characterized by the generalized growth of θ-oscillations and local (in the frontal leads) – α1- and β2-activities. Thus, in both groups of women surveyed the increase in activity of those cortical structures that ensure its implementation – sensory perception, motor start and motor programming commands and sense-motor coordination, was found. When MM are performed by women with a higher output α-frequency, they were characterized by more local changes in the electrical activity of the cerebral cortex. People with a lower modal α-frequency were characterized by somewhat less specific and differentiated features of the cortex activation. Alternating movements of fingers performed by women with low IαF were associated with higher EEG θ-, α1-, α2-activities, generally in the cortex, and α3-, β- and γ-oscillations – in the posterior areas than in those with high IαF. Instead of this, a relatively lower power spectral EEG was recorded in the frontal leads of the range consisting of α3-, β- and γ-oscillations. Intergroup differences found by us may indicate a higher status regarding a specific cortical tone, its readiness for activity and control of information processes in people with high IαF in comparison with those who had low IαF. It is natural that different levels of features of such activation systems achieve specific outcomes in testees with a different starting IαF. Women with a higher IαF had better speed characteristics of nervous processes.

93-96

Authors: V. L. Sokolenko, S. V. Sokolenko

The objective of this research is to assess the relation of radioactive contamination degree to immune system functioning, in the absence or presence of additional potential immunosuppressants. To achieve the objective, during the period of 1995–2015 we examined 250 people, students of Cherkasy State University, who lived in the areas of enhanced radiation monitoring before. Also we evaluated the additional impact of the emotional stress caused by examinations on examined students. Indicators of cellular immunity were determined by immunophenotyping and dyeing using Romanowsky-Giemsa method. The level of immunoglobulins in blood serum was determined by radial immunodiffusion (Mancini method). The level of cortisol in blood serum was determined by immunoenzyme method. We have found that in absence of the emotional stress among residents of the areas contaminated with radionuclides, cortisol level remained at the upper limit of homeostatic norm. There is an average positive correlation between the activity of radionuclides in the territories of residence and the level of cortisol. There are marked average positive correlations between the activity of radionuclides and the level of neutrophils, and low positive correlations with the levels of IgG and IgM in blood serum. Average negative correlations between the activity of radionuclides and the following parameters are also observed: absolute and relative number of functionally mature T-lymphocytes with phenotype CD3+, absolute and relative number of their helper subpopulation CD4+, absolute and relative number of natural killer cells with phenotype CD16+; and strong negative correlations with immunoregulatory index CD4+/CD8+. Cortisol level shows the similar correlation with the same parameters, but correlation coefficient is lower. Under conditions of additional stress, caused by emotional load during the examinations, cortisol level significantly increases. This enhanced previously discovered correlations and revealed new ones: average negative correlations between radionuclide activity / cortisol level and expression of pan-T cell marker CD5, medium positive correlations with absolute and relative number of suppressor T-lymphocytes with phenotype CD8+, correlation with the level of serum IgG changes from positive to negative. Additional short-term stress factor combined with chronic influence of low doses of radiation increases the dependence of immunosuppression, caused by radiation contamination of the areas where examined students live, and causes failures in the work of compensatory mechanisms, particularly at the level of serum immunoglobulin products.

103-107

Authors: H. N. Shiyntum, G. A. Ushakovа

Cadmiumclassified as a major carcinogen is considered a poisonous and unwanted heavy metal to a lot of tissues in many organisms. Of many publications already available, the general consensus is that the cadmium attenuating element is metallothionein (MT) through its interchangeable mechanism with Zn triggered by the presence of Cd, providing binding sites for Cd ions. MT was first discovered in the kidney cortex of the horse; it represents a low molecular weight protein, rich in cysteine residues which effectively bind with metals. Its functions consist in detoxification of heavy metals like mercury, arsenic, cadmium, homeostasis of essential metals including copper and zinc, anti-oxidation against reactive oxygen species, protection against DNA damage, oxidative stress, cell survival, angiogenesis, apoptosis, and increase of proliferation. In this work, we sought to highlight the protective function of MT in the brain and serum of rats by means of detoxification under induced effects of controlled Cd doses. We have done this by exposing Wistar rats to Cd at different doses in drinking water at different time intervals. In two independent experiments, 58 rats were subjected to 0.1 or 1.0 µg Cd2+/kg of body weight for 15 or 36 days under different conditions. The obtained data indicates the different functioning systems for the brain and the blood for MT metabolism under Cd effect. Our results indicate significant loss of metallothionein level in the brain and important increases in the amount of MT in serum proving that even minimal ingestion of toxic Cd is enough to trigger the release of MT protein in blood.

113-118

Authors: I. V. Prischepa, O. G. Prokushenkova, V. S. Nedzvetsky

Reactivation of glial cells, induced by metabolic disorders of glucose utilization and development of oxidative stress in retina under diabetes mellitus, is the key pathogenetic factor of diabetic retinopathy. Nanoparticles of C60 fullerene and some of their water-soluble derivates are known as one of the strongest antioxidants having neuroprotective effect in a number of pathologies and harmful influences. In the present study, for the first time, the effects of nanostructures of hydrated C60 fullerene (C60HyFn) on the expression and polypeptide composition of glial fibrillary acidic protein (GFAP) in retina of rats with streptozotocin (STZ)-induced diabetes have been evaluated. Using immunoblotting, 1.93-fold up-regulation of GFAP in diabetic rat retina as compared with control was shown, as a result of retinal glial cells reactivation induced by hyperglycemia. Increase in GFAP-immunolabeling associated with the reactive gliosis development in retina of diabetic rats was also confirmed by immuno-histochemical method. Consumption of C60HyFn solution (90 nM) as drinking water by diabetic rats for 12 weeks caused 1.51-fold decrease of GFAP level compared to untreated diabetic animals. In addition, C60HyFn caused statistically significant lowering of glycosylated hemoglobin concentration in blood serum of STZ-diabetic rats 1.58-fold. However, nanoparticles C60 did not affect neither insulin nor glucose levels in blood of diabetic rats. In conclusion, results obtained indicate that protective action of hydrated fullerene in the initial period of diabetic retinopathy of aged animals is realized through suppression of excessive activation of GFAP-positive retinal cells.

125-132

Authors: L. V. Altukhova, K. V. Kot, Y. G. Kot, K. S. Morozova, Y. E. Persky

Mechanisms of influence of volumetric autotransplantation of fibroblasts and of the mixture of fibroblasts and keratinocytes on the development of the local 3rd degree X-ray burn and the radiation skin ulcer in guinea pigs were investigated. We used deepadministration into the irradiation zone on its perimeter of 6 doses, which contained (150–160)×103 fibroblasts and (130–140)×103 keratinocytes in 100 µl. It is shown that this autotransplantation carried out 1 hour after the irradiation, and then every 24 hours, reduces the area of burn on the 35th day, compared to the control by 63%. Radiation ulcer appears on the 10th day after irradiation and is completely healed on the 25th day. With the same regimen of administration of only fibroblasts containing (200–210)×103 cells in 100 µl, these parameters of treatment were equal to 31% on 4th and 35th day, respectively. It is shown that as a result of radiation in the area of burn the level of gene expression of collagen types I and III, elastin, fibronectin, vinculin, decorin, hyaluronansynthases 1, 2, 3, matrix metalloproteinases 1, 2, 3, 7, 9 and hyaluronidase is reduced. Besides, in the burn area the level of gene expression of transforming growth factor α, fibroblast growth factors 1, 2, 8 and anti-inflammatory cytokines – interleukin 10 and transforming growth factor-β1 – is reduced, while the level of gene expression of proinflammatory cytokine (interleykin1β) increases. Both types of autotransplantation cause the growth of the expression level of all the structural genes and regulatory proteins of biopolymers and decrease in the expression level of interleukin 1β, which leads to activation of tissue regeneration and healing of the burn wound. Reasonsfor the higher efficiency of autotransplantation using the mixture of fibroblasts and keratinocytes compared to autotransplantation by fibroblasts only are both the larger total number of live cells regularly replacing dead cells in the burn area, and mutual stimulation of auto-fibroblasts and auto-keratinocytes to proliferate and to synthesize biologically active substances, i.e. cytokines and growth factors.

141-145

Authors: G. S. Maslak

Tumor growth progression of blood cells occurs due to changes in their genetic apparatus, which affects not only the cells morphological characteristics, but also their functional activity which to a greater extent depends on the membrane surface structures, a significant part of which is of glycoprotein nature. Complex type N-glycans are components of surface glycoproteins in the most of leukocytes. Thus, the study of changes in carbohydrate determinants of glycoproteins on the surface of leucocytes in tumorigenesis can help to reveal the mechanisms of this process. The aim of our study was to investigate the monocytes and granulocytes cytoplasmic membrane N-glycosylation in patients with chronic leukemia. The object of the study were blood cells of patients with chronic lymphocytic leukemia (n = 12) and polycythemia vera (n = 15) aged 58–66 years. Healthy hematologic volunteers (n = 15) aged 55 to 65 years were in the control group. N-glycan exposure on monocytes and granulocytes was investigated by flow cytometer Beckman Сoulter EPICS with Canavalia ensiformis lectin – Con A conjugated with fluorescent labels. The number of dead cells was monitored by means of binding them with propidium iodide. The result has been analyzed with FC Express. According to our data, levels of ConA-positive monocytes and granulocytes were 9,9 ± 1,0% and 32,7 ± 3,2%, respectively, in peripheral blood of healthy persons. The level of ConA-positive monocytes decreased to 31,0 ± 2,3% and the number of ConA-binding granulocytes increased to 66,7 ± 3,8% in patients with chronic lymphocytic leukemia compared with the norm. The number of ConA-positive monocytes decreased 3.3 times, and the level of granulocytes interacting with Canavalia ensiformis lectin slightly increased relative to control in polycythemia vera patients. There is significant increase in Con A-positive epitopes on granulocytes in patients with chronic lymphocytic leukemia and polycythemia vera compared with the control group, and absence of any difference in Con A-positive epitopes on monocytes. 100 times increase of the number of ConA-binding glycotopes is observed on the surface of granulocytes in peripheral blood of patients with polycythemia vera, and 3,3 times increase in patients with chronic lymphocytic leukemia, which may serve as an additional marker for the diagnosing these diseases.

151-155

Authors: D. Z. Vorobets

Methods of estimation of effectiveness of the open and laparoscopic pyeloplasty, as well as endo-urological palliative methods – laser resection, balloon dilatation and endopyelotomy, which determine the anatomical and functional peculiarities of renal pelvis and pyelo-ureteral junction with the help of ultrasound diagnostics during the forced diuresis, have been proposed. Changes of the area of renal pelvis, the velocity of post-furosemide increase of the scope of renal pelvis, rate of its drainage, changes in the diameter of pyeloureteral junction have been studied. This methodical approach is non-invasive, informative and simple in application. It is shown that dispersions of samples of patients after the open surgery do not differ from the dispersions of samples of the same patients before the operation on such parameters as areas of renal pelvis before the induction of furosemide, areas of renal pelvis after 15 minutes administration of furosemide, the rate of drainage after furosemide, the original diameter of pyeloureteral junction. This may indicate the stability of surgery results. For example, the larger renal pelvis by kidney size before the operation corresponded to the larger designed pelvis after the operation; renal pelvis drained faster before the operation, features faster drainage after the operation as well. Variation in the areas of renal pelvis which decreased in 40 minutes after furosemide, percent rate of longitudinal pelvis area, rate of after-furosemide increase in pelvis area, diameter of pyeloureteral junction in 15 minutes administration of furosemide after the open pyeloplasty was significantly different compared to the variation in the same parameter for the same patients before the operations. More substantial difference was observed in the same patients before and after Anderson-Hynes surgery by parameters of relative rate of after-furosemide drainage of pelvis and increase in diameter of pyeloureteral junction after 15 min administration of furosemide. That is, the same principle of operation provides similar results by anatomical parameters, such as size and diameter of pyeloureteral junction, but quite different results by functional parameters which reflect the possibility of draining of kidney in forced diuresis. Successful open pyeloplasty leads to a significant decrease in the pelvis area at different time intervals after furosemide administration, the relative increase in the pelvis area on the background of the induction of diuresis, rate of pelvis drainage, increase (normalization) in diameter of pyeloureteral junction, including the larger (better) gap of pyeloureteral junction after administration of diuretic. Concerning laparoscopic pyeloplasty, the dispersion of mean values of S, SPR, Vpr, V, VOT, D, DD after the operation was significantly different from those before the operation. This means that as in the case with open surgery, satisfactory clinical results such as reduction in renal pelvis and restoring the passage of urine through sufficient diameter pyeloureteral junction after laparoscopic pyeloplasty lead nevertheless to significant differences in the digital parameters during the objectification of operation effect by means of diuretic ultrasonography using furosemide-induced diuresis. Endoscopic surgery such as laser resection, endopyelotomy and balloon dilatation stably provides similar results (equal variances) by such parameters as pelvis area, which decreases in 40 min after furosemide administration, formation of the wide enough diameter of pyeloureteral junction and its minor fluctuations with the diuretic load. The decrease to normal parameters of all planes of renal pelvis (both before and after loading) and significant improvement of pelvis drainage (parameters responsible for the functional state of kidneys and pyeloureteral junction) indicate the success of palliative surgery in elimination of the narrowing of pyeloureteral junction.

161-164

Authors: Y. G. Kot, E. V. Kot, E. S. Morosova, E. E. Persky, M. A. Gritsenko, N. I. Bulankina

We studied gene expression of five metallothioneins (MT 1-5), ubiquitin and protein p53 and their products in fibroblasts culture of the skin and lungs of white rats of different ages (2 weeks, 1, 3, and 24 months) and determined its (metallothionein 1-5 types, ubiquitin, p53) product quantity. All these proteins are protective ones, but perform their functions by using different mechanisms. Metallothionein bind, transport and excrete ions of bivalent metals, ubiquitin controls the cleavage of the defective and short-lived proteins in the proteasome, protein p53 controls apoptosis, thus ensuring the genome stability. The similarity of age dynamics of gene expression of ubiquitin and MT of cells of both sources has been shown – maximum at 3 months. Expression of p53 gene has a difference: both in the skin and lungs expression increases up to 24 months. Product quantity of p53 has a minimum in the skin at 3 months and remains constant; in the lungs, this value has a maximum at 1 month.

9-16

Authors: S. L. Popel, B. M. Mitckan, S. Z. Krasnopolsky, I. V. Melnik

The purpose of this paper is to study the morphological changes of neurocytes in spinal cord of rats in hypokinesia and subsequent physical loading. Studies were performed on 55 laboratory rats of Wistar line. Materials of the research were the anterior horns of the gray matter of L5-S2 spinal segments. Preparations stained by Nissl and Viktorov were examined histologically. Hypokinesia was modeled following on the author’s technique. It was established that during prolonged hypokinesia in neurocytes of spinal cord of rats morphological changes in cell size and shape of the motor nuclei of all segments under study have been recorded. The number of hypochromic, hyperchromic destructively unchanged and hyperchromic destructively altered neurocytes increase; shadow cells appears, as well as cases of satellitosis and neuronophagia. Decrease in of albumen synthetical neurocyte function has been recorded. Physical loading of the average aerobic capacity leads to normalization of structural and functional state of neurocytes and enhances the reparative processes, as evidenced by a number of positive changes in morphometric parameters: increase in the number of normochromic neurocytes and decreasing the number of hyper- and hypochromic neurocytes with destructive signs, absence of pyknotic forms. Morphological parameters of neurocytes and their nuclei after physical loading of average aerobic capacity do not differ from those in the control group of intact animals. In neurocytes of this group of rats RNA concentration increases by 12.6% compared to animals after prolonged hypokinesia. Neurocytes of spinal cord of rats after prolonged hypokinesia develop significant morphological changes which are characterized by emergence of a significant number of hyperchromic neurocytes with signs of destructive changes and shadow cells, as well as and hypochromic neurocytes with signs of destructive changes, reduction in size and change of shape of perikaryons of neurocytes and their nuclei. Morphological changes of neurocytes after prolonged hypokinesia are accompanied by violations of biosynthetic processes, as evidenced by RNA decrease in the cytoplasm of efferent neurocytes of spinal cord of rats. Physical loading of average aerobic capacity leads to normalization of structural and functional state of neurocytes and promotes the reparative processes suported by positive changes in morphometric parameters.

23-27

Authors: O. V. Kulibaba, S. M. Kozishkurt, O. O. Duzenko, I. L. Vovchuk, S. A. Petrov

The paper presents the results of determining the amount of reduced glutathione, glutathione reductase and glutathione peroxidase activity at transplantation of muscle tissue taken from rats from the same litter. During the work 2 surgical interventions were carried out: transplantation of muscle tissue taken from the same litter of rats, and operation without replanting. Transplantation was performed on nonlinear white male rats weighing 180–300 g. The animals were taken out of the experiment by passing electric current through the medulla oblongata. For donor transplants males from the same litter were used. In adult donor rats abdominal muscle tissue was extracted and transplanted to homologous tissue of the recipient rat. The same procedure was carried out with femoral muscle tissue. Operation without replanting was used to compare the effects of changes in the number of investigated parameters of the surgical exposure. Tissue not subject to any surgery served as a control. Parameters under study were determined on the first, third and seventh day after transplantation. Glutathione levels in tissues were determined byE. Butler, A. Dyubona, B. Kelly. Determination of glutathione reductase is based on the reduction of NADPH registration. Glutathione peroxidase activity was determined by accumulation of oxidized glutathione. Surgery is known to lead to expression of oxidative stress in the organs operated. Special role in antioxidant protection of the body is given to thiols. Expressed hydrophilic properties provide their high content in water fraction of cells and ability to protect biologically important molecules (enzymes, nucleic acids, hemoglobin) from oxidative damage. Property of thiol compounds is known to inhibit both fermented and not fermented free radical oxidation. For the first time it was found that transplantation of muscle tissue taken from animals from the same litter lead to significant reduction of glutathione in the femoral muscle of the recipient and abdominal muscle tissue of the donor and recipient and to significant increase in glutathione reductase activity in the femoral muscle tissue of the donor and in the abdominal muscle tissue of the donor and recipient.

32-35

Authors: O. Y. Galkin, Y. V. Gorshunov, V. F. Solovjova

To address a number of fundamental and applied problems in immunology, molecular and cellular biology and biotechnology it is necessary to obtain Fc-fragments of immunoglobulins. Fc-fragments may be used for studying of the effector functions of antibodies which are mediated by these areas. They are often used as an immunogen to produce anti-specie (based on so-called secondary antibody) conjugate in the development of serological tests for diagnostics (predominantly such conjugate based on monoclonal antibodies). The work is aimed to develop improved methods of obtaining and allocation of Fc-fragments of human IgA. To achieve this objective, optimization of hydrolysis of IgA with subsequent purification of Fс-fragments have been carried out. Improved method of obtaining Fc-fragments of IgA provides: papain hydrolysis of immunoglobulin in the environment of nitrogen for 4 h, allowing to achieve maximum output of Fc-fragments without their further degradation: isolation and purification of Fc-fragments of human IgA by one-stage gel filtration on sephadex G-100; control of purity of the target product by electrophoresis in polyacrylamide gel with sodium dodecyl sulfate and Ouchterlony immunodiffusion. Enzymatic hydrolysis was carried out at the optimal temperature of papain (37 °C). As the oxygen in the air may have inhibitory effect on enzymatic hydrolysis reaction, the reaction mixture was incubated in the nitrogen atmosphere to prevent inactivation of papain. To reduce the incident degradation of immunoglobulin molecules, papain hydrolysis was carried out without using an enzyme activator (cysteine). Usage of the proposed scheme allows obtaining Fc-fragments of human IgA of high purity. Outcome of Fc-fragments after all stages of purification was about 18% of the initial amount of IgA in the preparation. Molecular weight from Fc-fragments of human IgA was equal to approximately 70 kDa.

40-44

Authors: G. І. Zvir, O. М. Moroz, S. O. Hnatush

Objects of the study were sulfate-reducing bacteria Desulfovibrio desulfuricans ІМV К-6, isolated from Yavorivske lakе. This strain is kept in the collection of microorganisms at the Department of Microbiology of Ivan Franko National University. Bacteria were grown in the Kravtsov-Sorokin’s liquid medium with the following composition (g/l): Na2SO4 × 10H2O – 0.5, NaH2PO4 – 0.3, K2HPO4 – 0.5, (NH4)2SO4 – 0.2, MgSO4 × 7H2O – 0.1, C3H5O3Na – 2.0. The bacteria were grown for 10 days at 30 °C under anaerobic conditions. In order to study the sensitivity of the sulfate reducing bacteria to action of Uragan and Raundup herbicides, the cells of D. desulfuricans ІМV К-6 were grown at the concentrations of herbicides as follows: 0,28 mМ, 2,8 mМ (concentration recommended for use) and 5,6 mM. Biomass was determined by photometric method. Concentration of hydrogen sulfide in the culture medium was determined by photo-colorimetric method. Concentration of sulfate-ions in the medium was determined by turbidimetric method. Capacity of sulfate reducing bacteria D. desulfuricans ІМV K-6 to grow, reducing sulfates to hydrogen sulfide upon influence of Uragan and Raundup herbicides was studied. Accumulation of bacterial biomass in the control and upon influence of herbicides was the highest on the fourth-sixth day of cultivation, and after that the stationary growth phase began. It was shown that sulfate reducing bacteria upon influence of herbicides grew more intensively compared with the control. It was discovered that the level of biomass changed depending on the increasing concentration of Uragan or Raundup herbicides in the medium. Sulfate reducing bacteria D. desulfuricans ІМV K-6 could reduce sulfates to hydrogen sulfide in the presence of sulfates and organic compounds in the medium (dissimilatory sulfate reduction). Stimulatory influence of Uragan and Raundup on the dissimilatory sulfate reduction process of D. desulfuricans ІМВ К-6 has been discovered. The formation of hydrogen sulfide correlates with the usage of sulfatе ions. The capacity of sulfate reducing bacteria D. desulfuricans ІМV K-6 to grow, reducing sulfate ions to hydrogen sulfide upon influence of Uragan and Raundup may be caused by presence of inert components (sulfates) in these herbicides that can be used by microorganisms as electron acceptors during sulfate respiration.

51-56

Authors: I. U. Pismenetskaya, T. D. Butters

Free oligosaccharides (FOS) are unbound structural analogs of glycans of glycoconjugates. There are several sources of them inside the cell: 1) multistep pathways of N-glycosylation, 2) the cell quality control and endoplastic reticulum-associated degradation of mis-glycosylated and/or misfolded glycoproteins, 3) lysosomal degradation of mature glycoconjugates. Some of these FOS are the earliest indicators of potential glycosylation alterations that would be revealed in the course of the cell quality control and the endoplastic reticulum-associated degradation. Ischemia and hypertension cause stress of intracellular organelles leading to disruption of their functions. The main objective of the work was the characterization of free oligosaccharides (FOS) in plasma obtained from patients with cardiovascular diseases compared to those from healthy subjects to evaluate the potential of these compounds for diagnostics. Chromatographic profiles of FOS composed of 4–12 monosaccharides were obtained and analyzed for quantitative and qualitative differences between the samples. After plasma deproteinization and FOS purification the oligosaccharides were labelled with anthranilic acid (2-AA), separated into the neutral and charged with QAE Sephadex (Q25-120) chromatography and analysed using high-performance liquid chromatography (HPLC). Glucose unit values were determined following comparison with a 2-AA-labelled glucose oligomer ladder derived from a partial hydrolysate of dextran as an external standard. The data were collected and processed using Empower software. The charged FOS were digested with the sialidase from Arthrobacter ureafaciens. 2-AA – labelled free oligosaccharides from transferrin were used as an external standard for the structure decoding. The profiles obtained were compared with intracellular free oligosaccharides of known structures and with the glycan structures and their descriptions in the databases GlycoBase and EUROCarbDB. These approaches allowed predicting a range of glycan structures for each of the main peaks of HPLC profiles of plasma free oligosaccharides and managing ways for their future experimental analysis. In the case of cardiovascular disorders, HPLC profiles of FOS revealed a changing pattern of heterogeneity, depending on the severity of the disease. Three main enlarged glycan species in the netral fraction and one peak in the charged fraction distinguished the FOS of the patients from those of the healthy volunteers. It has been revealed that the neutral marker peaks were represented by polimannose glycans with 5–7 mannose residues and 1–2 residues of N-acetylglucosamine, and one of the major peaks of the charged fraction – by two-antennary complex N-glycan with two sialic acid residues. The study of free oligosaccharides of blood plasma is a new field of glycobiology allowing an evaluation of an organism state at the level of the cell organelle functional status and openning up broad prospects for finding early diagnostic and prognostic markers of cardiac insufficiency.

63-67

Authors: O. E. Shaul’ska, L. M. Diachenko, T. P. Nikolayenko-Kamyshova, A. I. Shevtsova

Destruction of malignant cell clones and search for markers of the effectiveness of chemotherapy in patients with hematologic malignancies is an urgent direction of research. The purpose of this paper was to study plasma of patients with various forms of blood tumor, the activity of trypsin-like enzymes, matrix metalloproteinases MMP2 and MMP9, content of α1-proteinase inhibitor and α2-macroglobulin before and after the cytotoxic therapy by anthracycline antibiotics, doxorubicin and daunorubicin. It was established that concentration of inhibitors and activity of trypsin-like enzymes increased in patients with acute myeloid leukemia and multiple myeloma against the backdrop of multidirectional changes of proMMP9. Activity of the latter enzyme was reduced to 0,03±0,01 rel. u. in acute myeloid leukemia, and after chemotherapy it increased 7 times. Reducing level of inhibitors and increasing activity of MMP9 were found in chronic lymphocytic leukemia. The presence of the correlation between the α2-macroglobulin and gelatinase activity in the treatment indicates that the inhibitor is an important mechanism for storing of gelatinоlytic potential. Balance between proteolysis and antiproteolysis depends on the type of proliferating cells and the course of the disease. Indicators of tissue destruction may be an additional criterion for monitoring the stage of blood tumors and efficacy of treatment.

90-93

Authors: O. S. Voronkova, O. A. Sіrokvasha, T. N. Shevchenko, A. І. Vіnnіkov

Today film-forming strains of bacteria play very important role in clinical pathology. Staphylococci are ones of most dangerous of them. This bacteria can determine different pathological processes, for example, complication of airway mucosa. The ability to form a biofilm is one of the main properties of nosocomial strains. These strains should be monitored and their carriers are to be properly treated. To determine the origin of staphylococci strains we used bacteriophages from the International kit. The aim of research was to determine the phage type of staphylococci film-forming strains, that were isolated from naso-pharingial mucosa. Phagetyping has been carried out for 16 film-forming strains of S. aureus. To solve this problem, we used the International phage kit by Fisher’s method. As a result, sensitivity to phages from the International kit showed 53.8% of studied strains of S. aureus. 64.3% of sensitivity strains were lysed by one of the phage, 21.4% – were by two of the phages, 14.3% – by three of the phages. Isolates were sensitive to phages: 81 – 42.9%, 75 – 35.7%, 28.6%were sensitive to phages 47 and 53. All cases of detection of sensitivity to phage 47 coincided with the ability to form biofilm. Among non-film-forming strains there was no sensitive strains for this phage. Film-forming strains resist to erythromycin (62.5%), ciprofloxacin (43.8%), gentamicin (56.3%), tetracycline (87.5%), amoxicillin (93.8%), and cefuroxime (37.5%). All cases of sensitivity to phage 47 coincided with resistance to erythromycin, amoxicillin and tetracycline. For two of these strains,wealso defined resistance to gentamicin and for one of them – to ciprofloxacin. Results of research allowed to relate the bacterial cultures for determining the type. This may have implications for studying of film-forming ability, because surface structures of bacterial cell take place in this process. Belonging of an isolate to specific phage type may indicate possible differences in the structure of the cell wall, composition of the cytoplasmic membrane, receptors, etc. So, phage typing may be used as one of teststo study film-forming processes. Besides, phage typing can be helpful in finding of the source and modes of transmission of nosocomial strains of bacteria. Given the increased level of antibioticresistance among film-forming strains of staphylococci, the importance of this monitoring is indisputable, because spreading of these strains may cause the failure of therapy.

99-103

Authors: G. S. Maslak

Tn-antigen (Thomsen-nouvelle antigen) is tumor-associated carbohydrate antigen with only one GalNAc residue attached to serine or threonine of polypeptide chain. There is not enough data about the expression of this glycotope in hematologic processes. But the correlations between increasing Tn-antigen expression on the cell surface and tumor growth progression, invasion, and activation of cell migration are well known. Therefore, the currently important area of modern research is studying of the impact of anticancer therapy by expression of this carbohydrate antigen in the onco-proliferative process. There are two types of cytostatic therapies in clinical hospitals of Ukraine: COP-therapy (cyclophosphamide, vincristine, prednisone) and FC-therapy (fludarabine, cyclophosphamide), which are the most popular due to their effectiveness and low price. The aim of our study was to investigate Tn-antigen exposure on the surface of lymphocytes, monocytes and granulocytes in polycythemia vera and subleukemic myelosis; to examine the influence of COP- and FC-therapies on Tn-antigen exponation in patients with chronic lymphocytic leukemia. The objects of the study were blood cells of patients with chronic lymphocytic leukemia (n = 25), polycythemia vera (n = 15) and subleukemic myelosis (n = 15) aged 58–66 years. Healthy hematologic volunteers (n = 15) aged 55 to 65 years were in the control group. Lymphocytes of patients with chronic lymphocytic leukemia (n = 25) were also studied after the chemotherapy treatment of patients divided into two groups: those who took COP-therapy (n = 13); and those who treated with FC-therapy (n = 12). Tn-antigen exposure on lymphocytes, monocytes and granulocytes was investigated by Beckman Сoulter EPICS flow cytometer with primary monoclonal Tn-antigen anybodies (Institute of Immunology, Moscow, Russia) and secondary fluorescein isothiocyanate labeled antybodies (Millipore, USA). The number of dead cells was monitored by binding them with propidium iodide. The result was analyzed with FC Express. According to our data, Tn-antigen exposure was not detected on the surface of blood cells (lymphocytes, monocytes and granulocytes) in the control group and in patients with polycythemia vera and subleukemic myelosis. Nevertheless, Tn-antigen was identified on the surface of more than 80% of lymphocytes in chronic lymphocytic leukemia patients. The intensity of this tumor-associated antigen exposure on lymphocytes membrane was 100 times higher compared with that in normal lymphocytes. In chronic lymphocytic leukemia patients after COP-treatment the number of lymphocytes with surface Tn-antigen was equal to 28,1 ± 0,8%, and after FC-treatment it decreased to 9,5 ± 0,5%. Moreover, positive effect of cytotoxic therapy used in treatment of patients with chronic lymphocytic leukemia on intensity of Tn-antigen exposure on the surface of lymphocytes was shown. FC-therapy (fludarabine, cyclophosphamide) is more effective; compared with the data prior to this treatment it 40 times reduced the relevant index. Therefore, it can be applied in Ukraine for chemotherapeutic treatment schemes effective against Tn-antigen.

110-114

Authors: A. H. Yaderna, L. P. Golodok, A. I. Vinnikov

The method of polymerase chain reaction (PCR) in real-time was used to analyze the quantitative characteristics of normal and potentially pathogenic aerobic/facultative-anaerobic and anaerobic biota in the urethra, cervical channel and vagina in healthy women aged 10–40. The biota of all the women under 40 years and some of women older than 40 was mostly represented by lactobacilli. Microbialcomposition of the biocenosis in some women older than 40 is characterized by reduction in quantity of lactobacilli and their replacement by anaerobic microorganisms, mainly, such as Atopobium vaginae (16%), Gardnerella vaginalis (12%), Megasphaera spp. (8%), Dialister spp. (8%), Eubacterium spp. (8%) and Porphyromonas spp. (4%). Rarely, a wide range of other pathogens plays its role, including inter alia: Trichomonas vaginalis (8%), Mycoplasma genitalium (4%), Neisseria gonorrhoeae (2%) andChlamydia trachomatis (1%). The most frequent are the following strain associations: A. vaginae andG. vaginalis,Eubacterium spp. andPorphyromonas spp.,U. (urealyticum + parvum) andM. genitalium,N. gonorrhoeae andCh. trachomatis,U. (urealyticum + parvum) andCandida spp. This is connected with anatomical and physiological characteristics of genitals, hormonal and immune system action. Quantitative study of the biota of urogenital tract in Dnipropetrovsk women with the use of real-time PCR is the sensitive method for diagnosing both physiological and pathological changes, and dysbiotic disorders at early stages and preventing their further development into more serious forms.

121-124

Authors: E. Tkach, M. Vakerich, V. Nikolaychuk

Specificneed in manganese is a characteristic feature of all plants. At present time, participation of this element in functioning in a number of processes and structures in the plant organism is proved. Indispensabilityof manganese as a component of plant metabolism is evidenced by death of plants aged 2–4 weeks at absolute exclusion of manganese nutrient medium, and symptoms of disorders at all levels of metabolism due to lack of this element. As opposed to symptoms of deficiency which are specific for every element, the signs of excess are more or less the same. In conditions of gradual increase of the concentration of ions as factors of influence, gradual manifestation of plant organism depression is observed: growth inhibition, root dying and leaf chlorosis, etc. Foliar feeding of grain crops including winter wheat with manganese is very important because they have a great need for such elements as manganese and copper. Efficiencyof the types of foliar feeding is connected with the mechanisms of penetration of manganese into the plant cells. Unlike other metals, manganese can be mobilized by sprouts from seed integument: metal absorption from wheat seeds lasts till the seventh day. That’s why, the aim of our work was to investigate the influence of pre-sowing processing of winter wheat seeds with solutions of different concentrations of manganese sulfate for detection of the optimal concentration and studying of phytotoxic influence of the excess manganese action. Vegetation tests were carried out in the laboratory conditions for performing the studies. Winter wheat seeds were steeped into solutions of manganese sulfate in such concentrations as 0.001%, 0.01%, 0.1%, 0.5% and 1%. After that they were couched in multi-component environment (sod-podzol soil). As a control variant, an experiment without treatment was used. The influence of different concentrations on the growth processes such as seed germination, length of the sprout, weight of the plants and their root system was estimated. It was determined that the optimal concentration of manganese sulfate for the pre-sowing processing was equal to 0.001–0.01%. The critical one, at which the phytotoxic action is manifested in growth depression and decrease of sprout biomass, is the use of 0.5–1.0% solution of manganese sulfate for the pre-sowing seed treatment.

131-137

Authors: V. Borisenko, S. Belov, I. Sorokina, N. Gorgol

The research is aimed at the study of dynamic pathomorphological changes of choledoch and acute cholangitis development factors determined during the experiment. 36 rats of Wistar line were under trial. The main group consisted of 30 animals undergoing the open laparotomy, choledoch ligation and puncture modeling of acute cholangitis by E. coli culture in 1 х 108 CFU/ml concentration under general anesthesia. 6 healthy rats were included in the control group. Samples of general biliary duct under autopsy for pathomorphological study were taken on the 3rd, 7th, 14th, 21st and 30th day. In panoramic samples colored by hematoxilin and eozin the degree of dystrophic, necrobiotic, hemodynamic, inflammatory and atrophic manifestations’ changes were studied. Average depth of choledoch wall and height of its epithelial lining were morphometrically estimated. Collagen of the IV type as well as expressing receptors to CD34 were defined with the help of monoclonal antibodies in choledoch epithelial cells of basal membranes and choledoch vessels endotheliocytes. In choledoch, enhancement of edema and inflammatory infiltration by lymphoplasmocytic elements with the admixture of neutrofils with granulation tissue was detected from the 3rd up to the 30th day of the experiment. From the 14th day formation of bile clots of blood was detected in choledoch clearance, part of which was locked to its de-epitheliolized internal surface. According to morphometrical study data, choledoch wall depth increased from 261.1 ± 3.13 µm on the 3rd day to 572.5 ± 3.42 µm on the 30th day of the experiment. Mucosa membrane has lost its folding on the 14th day, epitheliocytes flattening was replaced by their destruction with fragments rejection into the duct lumen by the 30th day of the experiment. The epithelium height index decreased from 14.8 ± 0.09 µm on the 3rd day to 11.7 ± 0.15 µm on the 30 day of the experiment. Collagen of the IV type fluorescence intensity of vessel basal membranes from the 3rd day closely matched the control, its enhancement was detected by the 30th day. Choledoch epitheliocytes fluorescence, expressing the receptors to CD34 reliably decreased by the 30th day of the study. In its turn, content of collagen of the IV tyre, as well as vessels endothelium expression in preparations treated by PQA to CD34 reliably increased by the 30th day of the experiment. Thus, in pathogenesis of acute cholangitis apart from well-known factors such as cholestasis and infection the third factor was detected, in the way of biliary ducts mucosa membrane lesion. In morphogenesis of choledoch mucosa membrane lesion the decrease in role of epitheliocytes adhesive properties was stated as well as deficit of collagen of the IV type in the structure of epithelial basal membranes. During observations where epithelial covering consistency was preserved, inflammatory changes in choledoch were insignificant, which is proved by sufficient resistance of biliary ducts epithelium to infection in the presence of cholestasis and bacteriocholia, but in cases when mucosa membrane de-epitheliolization took place the development of severe purulent-destructive cholangitis and pericholangitis was detected.

143-147

Authors: S. Kyrychenko, I. Prishchepa, V. Lagoda, M. Velika, V. Nedzvetsky

The aim of this study was to examine whether the antioxidant alpha-lipoic acid protects neurons from diabetic-reperfusion injury. The streptozotocin (STZ) rat model was used to study the glial reactivity and prevention of gliosis by alpha-lipoic acid (alpha-LA) administration. The expression of glial fibrillary acidic protein (GFAP) was determined, as well as lipid peroxidation (LPO) and glu-tathione (GSH) levels in some brain tissues. We observed significant increasing of lipid peroxidation products in both hippocampus and cortex. Changesof polypeptide GFAP were observed in hippocampus and cortex. Both soluble and filamentous forms of GFAP featured the increase in hippocampus of rat with hyperthyreosis. In the filamentfractions, increase in the intensity of 49 kDa polypeptide band was found. In the same fraction of insoluble cytoskeleton proteins degraded HFKB polypeptides with molecular weight in the range of 46–41 kDa appeared. Markedincrease of degraded polypeptides was found in the soluble fraction of the brain stem. The intensity of the intact polypeptide – 49 kDa, as well as in the filament fraction, significantly increased. It is possible that increasing concentrations of soluble subunits glial filaments may be due to dissociation of own filaments during the reorganization of cytoskeleton structures. Given the results of Western blotting for filament fraction, increased content of soluble intact 49 kDa polypeptide is primarily the result of increased expression of HFKB and only partly due to redistribution of existing filament structures. Calculation and analysis of indicators showed high correlation between the increase in content and peroxidation products of HFKB.These results indicate the important role of oxidative stress in the induction of astroglial response under conditions of diabet encefalopathia. Administration of alpha-LA reduced the expression both of glial and neuronal markers. In addition, alpha-LA significantly prevented the increase in LPO levels found in diabetic rats. GSH levels increased by the administration of alpha-LA. This study suggests that alpha-LA prevents neural injury by inhibiting oxidative stress and suppressing reactive gliosis. All these changes were clearly counteracted by alpha-lipoic acid. The results of this study demonstrate that alpha-lipoic acid provides for protection to the GFAP, as a whole, from diabet -reperfusion injuries.

7-11

Authors: O. I. Sidashenko, O. S. Voronkova, Y. A. Sirokvasha, A. I. Vinnikov

122 strains of staphylococci were identified. Among the examined 122 clinical strains of staphylococci, 67 strains belonged to coagulase-positive, and 55 strains to the coagulase-negative ones. According to the study of physiological and biochemical properties, it was found that 37 strains (30.3%) belonged to S. epidermidis species. One of the biological properties of many bacteria is the ability to film formation and these strains attract special attention, since it is known that the film antibiotic resistance is higher than in planktonic cultures. It was determined that 20 strains of those under study were film-forming, 17 strains – non-biofilm forming ones. The film was formed during three days, and settled to the bottom of the plate holes. The clinical (Cl) strain of S. epidermidis was sensitive to ceftriaxone and tetracicline. The control (C) strains of S. epidermidis were sensitive to ceftriaxone, tetracycline and sizomicine. The study of biofilm growth for 2, 3 and 4 days of incubation was carried out. The maximum rate of biofilm S. epidermidis C was observed during 2–3 days; there is the most intense increase of cells number from 5.2 × 108 CFU/ml, for S. epidermidis Cl to 5.6 × 108 CFU/ml. The effect of ceftriaxone and tetracycline on biofilm formation by 2 investigation strains of S. epidermidis was found. We determined differences in minimal inhibitory concentrations (MIC) for planktonic cultures and biofilm of strains under study. It was established that MIC antibiotics inhibited the growth of planktonic cultures on average 2 times lower compared to the MIC which inhibited the biofilm formation. MIC for planktonic culture of S. epidermidis Cl defined for ceftriaxone was equal to 10 mg/ml, and for tetracycline – 1 mg/ml. MIC of ceftriaxone for the control strain was equal to 12 mg/ml, MIC of tetracycline – 0.7 mg/ml. MIC values for dynamics biofilm formation of S. epidermidis Cl strain on the plater were as follows: to ceftriaxone – 20 mg/ml and for tetracycline – 2 mg/ml, MIC of ceftriaxone for S. epidermidis C strain – 24 mg/ml, MIC of tetracycline – 1.5 mg/ml. The effect of ceftriaxone and tetracycline was defined to the larger extent, than MIC for biofilm-forming on the plate (10, 50 and 100 times). More effective action of tetracycline was shown for 1- and 2-daily biofilm cultures of S. epidermidis clinical strain. Adding tetracycline concentration of 20 mg/ml in the culture medium of the 1-day biofilm of S. epidermidis Cl strain reduced the cell number of biofilm formation 590 times, increasing concentrations of tetracycline to 100 mg/ml and added to the 1-day biofilm of the clinical strain reduced the number of bacterial cell 4400 times compared with control.

17-22

Authors: O. V. Chaplia, J. V. Gontar, N. M. Bilko

In order to reveal the influence of genetic component on the early embryo development, the retrospective study of morphokinetic characteristics of 717 embryos subjected to preimplantation genetic testing was conducted. Blastomere biopsy for FISH-based preimplantation genetic screening of 7 chromosomes was performed on the third day of culture, while embryo developmental potential and morphological features at the cleavage and blastulation stage were studied regarding maternal age particularly in the group of younger women and patients older than 36. Results of genetic testing revealed that euploid embryos rate gradually decreased with maternal age comprising 39.9% in young women group and 25.3% of specimen belonging to elder patients. At the cleavage stage, morphological characteristics of aneuploid and euploid embryos didn’t differ significantly regardless of the age of patients that could be accounted for the transcriptional silence of embryo genome till the third day of its development. However, in case of prolonged culture chromosomally balanced embryos rarely faced developmental arrest (in 7.9%) and formed blastocysts half more frequently compared to aberrant embryos (respectively 75.6 versus 49.8%). Nevertheless, no substantial difference was found between blastocyst formation rate among embryos with similar genetic component regardless of the maternal age. Taking into consideration high rate of chromosomally unbalanced embryos specific to patients of advanced maternal age, the relative proportion of aneuplouid blastocysts was significantly higher in this group of embryos. Thus, without genetic screening there is a possibility of inaccurate selection of embryos for women of advanced reproductive age for transfer procedure even in case of prolonged culture. Consequently, increase of aneuploid embryos frequency associated with permanent preimplantation natural selection effectiveness along with the postimplantation natural selection failure may be the cause of elevated risk of chromosomally abnormal child birth for advanced age women after natural conception, as well as after assisted reproduction therapy

28-32

Authors: V. L. Sokolenko, S. V. Sokolenko

The aim of this study is to determine the role of haptoglobin phenotype in realization of adaptive responses of cellular and humoral immunity indices to moderate exercise caused by physical training. The study was implemented in the group of second-year students aged 18–20 who lived in the same climatic and geographical conditions for a long period of time. The students didn’t have any acute or chronic diseases and attended the main group of physical training. 60 persons were investigated. Immune system indices analysis was carried out in September before and after physical training lessons. Leukocyte level was calculated using hemocytometer, lymphocyte level was determined on the base of blood smear (dyeing for Romanowsky–Giemsa). Expression of surface antigene by peripheral blood lymphocyte was determined by immuno-fluorescence method with the use of monoclonal antibodies. The level of immunoglobulin in plasma was determined by radial immunodiffusion or Mancini method. To assess the phenotype of haptoglobin (Hp) we used the method of electrophoresis in starch gels. In the course of research we have detected the reduction of the relative and total number of lymphocytes regardless of haptoglobin phenotype in the group of students after physical training; this is a typical feature of the initial stages of stress response. We observed statistically reliable decrease in total number of analyzed subpopulations of T-lymphocyte in the group of students with phenotype Hp2-2 which was obviously the result of changes in the general level of lymphocytes in the peripheral blood. In the group of students with phenotype Hp1-1 absolute number of T-lymphocyte with phenotype CD3+ and CD4+ is reduced. In the group of students with phenotype Hp2-1 we have seen only the tendency to decrease in functional mature T-lymphocyte and their helper subpopulation. In the group of students with phenotype Hp2-2 the relative number of helper T-lymphocyte with the phenotype of CD4+ was reduced after moderate physical activity and it caused the decline in immune regulating index CD4+/CD8+. Changes of the level of B-lymphocyte with CD72+ phenotype were not found regardless of the haptoglobin phenotype. Analysis of the impact of moderate physical activity on major classes of serum immunoglobulin revealed the reduction of IgG level in the group of students with phenotype Hp2-2. Thus, we draw the conclusion that serum haptoglobin system shares the adaptive reactions of immune system to moderate physical activity. The most pressing phenotype is Hp2-2. Even in the group of students with immune pressing haptoglobin phenotypes, the analyzed parameters do not go beyond the homeostatic regulations after physical activity and it indicates satisfactory level of immune system adaptation to performed physical activity.

39-44

Authors: V. V. Avakov, N. N. Rozhko

The influence of the environment on the child health is one of priority issues of the present time and it is of great social importance. Increased dental diseases associated with climatic and geographical characteristics of the area are widely discussed in the literature. The leading among them are environmentally determined dental diseases in children associated with geochemical and technogenic pollution of the area where they live. Increasing amounts of hard metal salts is the urgent hygienic problem, due to severity of their multi-element analysis in microsubjects, and negative influence on health of the children population, due to tropism, ability to cumulation, long biological life in the body and antagonism of heavy metal salts to the number of microelements. Influence of hard metal salts on dental diseases development is undeniable. Particular attention is paid to their influence on caries process and treatment peculiarities. Despite the fact that modern аdhesive dentistry in recent years has made a significant breakthrough in improving adhesive systems, correct choice of adhesive system depending on changes in the structure of hard tissue under geochemical contaminants (like heavy metal salts) is the most important step. It is the decisive factor for adaptation and connection of restoration with the restoration base. We should remember that on the way of adhesive system there is an altered structure preventing from deep penetration of such system and, consequently, leading to violation of restoration tightness. Therefore, early detection of complications by clinical evaluation of quality of the restorations is of great interest. Multi-vector approach to treatment of dental caries in children living in conditions of technogenic pollution by heavy metal salts is extremely urgent and important issue. Significant niche in this approach is given to adhesive preparation methods combined with local fluoridation, using fluoride medication of the new generation. One of them is Hluftored which promotes the formation of microcrystalline СаF2. Protocol of work with this new generation drug is based on the sequential precoating of enamel with magnesium-fluoride silicate, and then with suspension of high desperasible calcium hydroxide promoting formation of microcrystalline СаF2, magnesium and copper with the size of ~50Аº, leading to easy penetration of fluoride into pores of softening range. Moreover, microcrystals of CaF2 are protected by silicic acid gel from outwashing. Therefore, they release fluoride for a long period of time (over 1 year), which promotes remineralization and prevents complications of dental caries. This article includes the data regarding effectiveness of the Hluftored use in combination with various adhesive systems. Also, we show data of clinical quality assessment conducted for restoration procedures in children of different age groups and territories by using index of USPHS, after 6 and 18 months of carrying out such procedures. Clinical observations indicate the effectiveness of deep fluoridation and different adhesive systems in the complex of caries treatment in different age groups at present time and in future.

49-53

Authors: A. Y. Varigin

The degree of phenotypic plasticity in the Idotea baltica basteri Audouin, 1827 (Crustacea, Isopoda) sex ratio of the fouling community in the Odessa bay of the Black Sea was determined. The ratio of males and females in three main phenotypes of I. baltica basteri, namely: uniformis, albafusca and lineata was found. Crustaceans were collected on the underwater surface of traverses, located in three sea districts with the different degrees of water exchange intensity with the open sea. Water depth at the wall of traverses ranged from 1 to2,5 m. In the first most opened district among the individuals of uniformis phenotype the ratio of males and females was equal to 1:4, that for albafusca – 1:1,3 and lineata – 1:1. In the second area with the release of drainage waters among the individuals of uniformis phenotype this ratio was equal to 1:4,5, while the albafusca and lineata figures were 1:1,5 and 1:1, accordingly. In the third region, with the most difficult water exchange, among the individuals of phenotypes the ratios of males and females were as follows: uniformis – 1:8, albafusca – 1:2, and lineata – 1:1. When moving from the opened to semi-enclosed area among the individuals of all phenotypes there was a gradual decline in the proportion of males and accordingly, increases of the proportion of females. The greatest number of males in all areas under study is observed among the individuals of lineata phenotype, and that of females – among the individuals of uniformis phenotype. It is found that monochromatic colored females prefer to stay in the shaded places among the seagrass beds and brightly colored males usually move actively along the outside of the substrata.

58-61

Authors: H. M. Lisunets, V. S. Nedzvetsky

The results of experimental investigation of autoimmune reaction targeted against neirospecific protein antigens in patients with congenital heart failure are presented. It is shown that titer for autoanibodies to these patients’ brain proteins is significantly higher compared with healthy persons. Elevation of titer of these autoantibodies is correlated with the ability to solve tasks for revealing complex analogies and decrease of life quality. Therefore, worsening of life quality and development of cognitive deficit in patients with cardiovascular diseases might be associated with autoimmune reaction against specific proteins of nervous tissue cells. The results obtained demonstrate that generation of autoantibodies in patients with congenital heart failure is linked with the cognitive deficit. One of the most important causes of cognitive decline could be hypoxia state due to surgery intervention. Consequences of hypoxia, in their turn, are accompanied by chronic oxidative stress. Molecular and cell injuries induced by free radicals are known to be the widely spread cause of various pathologies. Further investigations for validation of diagnostic hallmarks for estimation of the origin of progression of the cognitive deficit in patients with congenital heart failure are needed. These studies will allow clarifying mechanisms of influence of hypoxia-induced injuries associated with the aggravation of oxidative stress in nervous tissue cells. Understanding of relationship between decline of cognitive function and pathologies of cardiovascular system is necessary for more precise clinical diagnostics of the early prevention of possible complications and proper treatment of patients.

66-70

Authors: S. V. Kyrychenko, V. S. Nedzvetsky

The influence of hyperthyreosis on oxidative stress, state of glial intermediate filaments and memotry was investigated. Significant increasing of lipid peroxidation products into both hippocampus and cortex and change for the worse of memory was observed. Analysis of the behavioral reactions of rats in the test of passive avoidance conditioned reflex showed that the acquisition of skills of all groups of animals did not differ by time waiting period (latent period). Time saving memory test conditioned reflex of passive avoidance was excellent in the group of rats treated with thyroxine compared with controls. The change of polypeptide GFAP was observed in hippocampus and cortex. Both soluble and filamentous forms of GFAP increased in hippocampus of rat with hyperthyreosis. In filament fractions, increase in the intensity of 49 kDa polypeptide band was found. In the same fraction of insoluble cytoskeleton proteins degraded HFKB polypeptides with molecular weight in the region of 46–41 kDa appeared. Marked increase of degraded polypeptides was found in the soluble fraction of the brain stem. The intensity of the intact polypeptide (49 kDa), as well as in the filament fraction, significantly increased. It is possible that increasing concentrations of soluble subunits glial filaments may be due to dissociation of own filaments during the reorganization of cytoskeleton structures. Given the results of Western blotting for filament fraction, increased content of soluble intact 49 kDa polypeptide is primarily the result of increased expression of HFKB and only partly due to redistribution of existing filament structures. Calculation and analysis of indicators showed high correlation between the increase in content and peroxidation products of HFKB. These results indicate the important role of oxidative stress in the induction of astroglial reactive response under conditions of hyperthyroidism. This data shows the possibility of the glial cell cytoskeleton reconstruction under effect of thyroid hormones.