THE INFLUENCE OF CHRONIC HYPERHOMOCYSTEINEMIA ON PHAGOCYTIC AND METABOLIC ACTIVITY OF PERIPHERAL BLOOD NEUTROPHILS IN CASE OF LIPOPOLYSACCHARIDE-INDUCED PERIODONTITIS.

The aim of the study was to investigate the phagocytic and metabolic activity of peripheral blood neutrophils in rats with lipopolysaccharide (LPS)-induced periodontitis combined with chronic thiolactone hyperhomocysteinemia (HHcy).The experiment included non-linear mature male rats (n=48), which were divided into 4 groups: control; animals with a periodontitis model; animals with a model of chronic thiolactone HHcy; animals with a model of periodontitis in combination with chronic thiolactone HHcy. Phagocytic activity, phagocytic index and phagocytic number were determined as indicators of phagocytosis of peripheral blood neutrophils. The oxygen-dependent bactericidal activity of peripheral blood neutrophils was studied using nitroblue tetrazolium test (NBT-test).Our research has found that LPS-induced periodontitis in rats is accompanied by dysfunction of phagocytosis process (increased phagocytic activity with a simultaneous decrease of absorption capacity) and activation of oxygen-dependent microbicidal mechanisms of peripheral blood neutrophils, as indicated by an increase of indices of spontaneous and activated NBT-test. Chronic thiolactone HHcy adversely affects the functional and metabolic activity of peripheral blood neutrophils in case of periodontitis, which is confirmed by a violation of the process of phagocytosis, a more pronounced decrease in absorption capacity and depletion of metabolic reserves of these cells in rats with comorbid course of LPS-induced periodontitis vs. animals with only LPS-induced periodontitis. The observed disorders in the process of phagocytosis in rats with comorbid course of periodontitis are an important factor in reducing the non-specific organism resistance which contributes to the progression of periodontitis. The obtained results reveal new aspects of the high Hcys plasma level influence on the course of inflammatory process in periodontal tissues, which opens opportunities for improving pathogenetic therapy in patients with periodontal disease combined with chronic HHcy.

[PECULIARITIES OF IMMUNE PROFILE IN BLOOD OF RATS IN THE DYNAMICS OF IMMOBILIZATION STRESS ON THE BACKGROUND OF HYPOTHYROIDISM].

The aim of our study was to investigate the peculiarities of changes in the indices of cellular and humoral immunity in the case of immobilization stress, which is implemented against the background of hypothyroidism. It was used 84 white Wistar male rats to study the peculiarities of the stress reaction against hypothyroidism. The study of cellular and humoral immunity was performed on a Beckman Coulter Epics-XL flow cytometer. During the immobilization stress in rats, the cellular immunity indices reliably decrease at the anxiety stage with adaptation to the effect of stress after 48 h in the resistance stage with subsequent exhaustion of the adaptation reserves at the depletion stage. Against the background of a decrease in the T-lymphocyte level at the anxiety stage, the B-lymphocyte population significantly increased (by 26.5%). In animals with deficiency of iodine-containing thyroid hormones, we noted a significant decrease in cellular and humoral immunity at all stages of immobilization stress, which was accompanied by a significant decrease of immunoregulatory index (p<0.05).

THE SPECIFIC FEATURES OF CELL DEATH OF CIRCULATING NEUTROPHILS IN A SETTING OF EXPERIMENTALLY INDUCED CRUSH SYNDROME.

Although the overall incidence of trauma-induced crush syndrome (CS) is as low as 0.1 per 10,000 of population, which makes it one of the least common traumatic injuries, the hazard of CS lies in the high incidence of hemodynamic and metabolic complications, which call for an early diagnosis of this syndrome. This paper reports the specific features of cell death of neutrophils in rat blood as they changed with time using the model of endogenous toxemia formed in a setting of crush syndrome. To assess apoptosis/necrosis in neutrophils ANNEXIN V FITC assay kit (manufactured by Beckmann Coulter, USA) was used. The results obtained in this study suggest an increase in apoptotic and necrotic processes in a setting of CS from the first day of the experiment through the end of observation. Also we have found a significant impact of reactive oxygen species generation on neutrophils death in a setting of CS. However, other mechanisms to launch apoptosis/necrosis in case of experimental crush syndrome should not be neglected.

THE INFLUENCE OF MONOSODIUM GLUTAMATE ADMINISTRATION ON GENERATION OF REACTIVE OXYGEN SPECIES AND APOPTOSIS OF BLOOD LEUKOCYTES IN RATS.

The use of food additives in food manufacturing has been a public health issue for many years. Currently, more than 3000 food additives have been approved and estimates has shown that each person may consume 3.6 to 4.5 kg of food additives per year on average. One of the most common food additives in Ukraine and in Europe is monosodium glutamate (MSG). This paper reports the effect of monosodium glutamate administration at "safe" (allowed) doses on the generation of reactive oxygen species and apoptosis in leukocyte blood suspension of rats. Experimental studies were conducted on 32 non-linear, male, white rats weighing 180-200 g. The experimental animals were administered MSG (Sigma-Aldrich, USA) at a dose of 15, 30 and 45 mg/kg body weight. Control group of animals received normal saline. We have found that consumption of MSG at low dosage was associated with development of excessive reactive oxygen species generation and apoptotic processes in blood leukocytes of rats. Therefore it is advisable not only to investigate the established dangerous doses of E621, but also to study the molecular mechanisms of the "safe" (allowed) doses of MSG effect on a living organism.

THE INDICES OF ENDOGENOUS INTOXICATION IN RATS WITH CARRAGEENAN SOLUTION CONSUMPTION.

With changing lifestyle and dietary transition, there is an increase in intake of processed and packaged foods which tend to have a number of food additives. This has increased our consumption of these chemical substances. One of such additives is carrageenan (CGN) - E407. This paper reports the effect of 0.5 % carrageenan solution consumption on the main indices of endogenous intoxication in rats. Experimental studies were conducted on 48 non-linear, male, white rats weighing 180-200 g. The experimental animals had free access to 0.5% carrageenan solution (Sigma-Aldrich, USA) in drinking water. Control group of animals received pure water. Syndrome of endogenous intoxication was evaluated using measurement of the middle mass molecules contents in blood serum. We have found that even the minimal intake of carrageenan, triggers the mechanisms of endogenous intoxication in rats, starting in 2 weeks of consumption. 1 month consumption of carrageenan with drinking water in concentration of 0.5% is associated with significant increase in endogenous intoxication, manifested by increased contents of middle mass molecules (both the chain amino acids and the aromatic amino acids) in blood serum. From these facts, one may conclude that although carrageenan has been used widely in food as an emulsifier, a stabilizer, and a thickener for more than 50 years, some questions of its safety are still opened.

INVESTIGATION OF BONE MINERALIZATION IN PATIENTS WITH CORONARY HEART DISEASE COMPLICATED BY CHRONIC HEART FAILURE, STAGE II-A.

The majority of the studies have shown that individuals with cardiovascular diseases have a higher risk of experiencing bone loss and thus greater predisposition to risk of fracture. On the other hand there is growing evidence that individuals with low bone mass have higher mortality for cardiovascular events compared to patients with cardiovascular disease with normal bone mass. This research aims to investigate bone mineralization in patients with coronary heart disease complicated by stage II-A chronic heart failure. The study involved 33 men with coronary heart disease complicated by Stage II-A chronic heart failure. Bone mineral density was measured using dual energy x-ray densitometry of lumbar region of spine. Structural and functional changes of bone tissue of the lumbar spine have been found in 49,2% patients with coronary heart disease complicated by Stage II-A chronic heart failure, in particular, I stage of osteopenia - in 44,6%, II stage of osteopenia - in 27,7%, III stage of osteopenia - in 10,8% and osteoporosis - in 16,9%. It was established the same type of downward trend for BMD decreasing in L1 of patients with different stages of osteopenia, but in case of osteoporosis mineralization decreased equally in all vertebrae.

THE DETERMINATION OF CORRELATION LINKAGES BETWEEN LEVEL OF REACTIVE OXYGEN SPECIES, CONTENTS OF NEUTROPHILES AND BLOOD GAS COMPOSITION IN EXPERIMENTAL ACUTE LUNG INJURY.

Acute lung injury (ALI) remains a major cause of acute respiratory failure and death of patients. Despite the achievements at the current stage in treatment, morbidity and mortality of ALI remain high. However, a deeper understanding of the pathogenetic links of ALI, identifying of the predictors that positively or negatively influence on the course of the syndrome, the correlation between some pathogenetic mechanisms will improve therapeutic strategies for patients with ALI, which makes the actuality of this study. The aim of the research was to detect additional pathogenetic mechanisms of the acute lung injury development in rats based on a comparative analysis of the correlations between the level of reactive oxygen species in blood and bronchoalveolar lavage, contents of neutrophils and blood gas composition. The experiments were performed on 54 white nonlinear mature male rats 200-220g in weight. The animals were divided into 5 groups: the 1st - control group (n=6), the 2nd - animals affected by hydrochloric acid for 2 hours (n=12), the 3rd - animals affected by hydrochloric acid for 6 hours (n=12), the 4th - animals affected by hydrochloric acid for 12 hours (n=12), the 5th - animals affected by hydrochloric acid for 24 hours (n=12). Correlation analysis was performed between all the studied indices. Coefficient of linear correlation (r) and its fidelity (p) was calculated that was accordingly denoted in the tables (correlation matrices). The correlation coefficient was significant at p<0.05. Conducted correlative analysis showed that the level of ROS in neutrophils of blood in rats with modeled ALI had a high negative correlative linkage with pH of arterial blood in 2nd and 3rd experimental groups. Conducted correlative analysis of data in BAL showed that the level of ROS in neutrophils in rats with modeled ALI had a strong positive correlative relationship with the number of white blood cells in 3-rd, 4-th and 5-th experimental groups and positive correlative relationship with the absolute number of neutrophils in the same experimental group. Consequently, the main manifestation of acute lung injury is the development of hypoxia and respiratory acidosis that is caused by the violation of gases diffusion in the alveoli. On this background non-specific inflammatory reaction is developed at lung microvessels level with violation of lung homeostasis, which is iniciated by neutrophils' activation, which are producing ROS.