Medical and Clinical Chemistry

INFLUENCE OF GLYCEMIA LEVELS IN PATIENTS WITH DIABETES ON THE STATUS OF THE MICROBIOME OF DIFFERENT BIOTOPES

L. I. Malinovska — 2023-02-28

Introduction. The incidence of diabetes mellitus, which is steadily increasing in the human population, and the complications that occur in such patients, caused by representatives of the autochthonous microflora, prompt the study of the reasons that contribute to such processes, in particular, the influence of glycated hemoglobin levels and changes in the pathogenic properties of microorganisms in patients with diabetes.

The aim of the study – to investigate literature data on changes in the structure and properties of the microbiome of certain biotopes, patients with diabetes, and to find out the influence of glycated hemoglobin levels on the properties of representatives of the microflora.

Research Methods. To carry out this review, we analyzed articles published in scientific journals. For the search, we used databases of leading foreign publications. Those articles containing information about the course of diabetes and the state of the microbiome of certain areas were chosen as the selection criteria.

Results and Discussion. The review presents and analyzes the results of domestic and foreign research, with references to primary sources, on changes in the structure and certain properties of microorganisms – representatives of the autochthonous microflora of the oral cavity, stomach, intestines, conjunctiva, the frequency of detection of pathogenic and conditionally pathogenic microorganisms in the above biotopes. Their impact on glucose metabolism, the impact of glycemic levels on biodiversity and virulence properties of microbiota, in particular S. aureus, Helicobacter pylori, Staphylococcus epidermidis and others, changes in the ratio of aerobic and anaerobic intestinal microbiota in patients with diabetes and when it is combined with other nosological forms and in experimental conditions.

Conclusions. Summarizing the results of the studied literature data, it can be stated that the level of glycated hemoglobin directly or indirectly affects the frequency of bacterial complications in patients with DM, namely by increasing the virulence properties of representatives of the normal flora of different biotopes, increasing the colonization of specific areas of the mucous membranes or skin by a certain microbe, and disrupting the relationship between different representatives of a separate biotope.

THE EFFECT OF METFORMIN AND ITS COMBINATION WITH MODULATORS OF HYDROGEN SULPHIDE METABOLISM ON THE LEVEL OF GLYCEMIA AND THE STATE OF THE H2S SYSTEM IN THE KIDNEYS OF RATS WITH STREPTOZOTOCIN-INDUCED DIABETES

O. B. Strutynska — 2023-02-28

Introduction. Diabetic nephropathy belongs to one of the severe microvascular complications of diabetes mellitus (DM) and is one of the causes of patient disability and mortality. An important role in the treatment of diabetic nephropathy belongs to the sugar-lowering drug metformin. The question of the molecular mechanisms of metformin’s action, in particular the role of the H₂S signaling system in its pharmacological activity, remains unclear.

The aim of the study – to evaluate the effect of metformin and its combination with modulators of H₂S, exchange on the level of glycemia and H₂S metabolism in the kidneys of rats with streptozotocin-induced diabetes.

Materials and Methods. The experiments were performed on 75 white non-linear male rats weighing 150–240 g. The animals were divided into five groups: group 1 – control; group 2 – animals with experimental DM, which was initiated by a single intraperitoneal injection of streptozotocin (40 mg/kg of weight) in 0.1 M citrate buffer (pH 4,5); group 3 – animals with experimental DM were treated with metformin (500 mg/kg/day, intragastrically) from the 3^rdto the 28^th day; group 4 – animals with DM along with metformin were given NaHS (56 μmol/kg/day, intragastrically); group 5 – animals with DM along with metformin, were administered propargylglycine (PPG, 442 μmol/kg/day, intragastrically). The glucose content was determined in the peripheral blood. H₂S level, activity of H₂S-synthesizing enzymes (cystathionine-γ-lyase – CSE, cystathionine-β-synthase – CBS, cysteineaminotransferase/3-mercaptopyruvate sulfurtransferase – CAT/3-MST), activity of thioredoxin reductase (TRR) and rate of H₂S utilization were evaluated in the supernatant of the kidney homogenate,

Results and Discussion. It was established that streptozotocin-induced diabetes (ST-diabetes) causes a significant increase in blood glucose levels (by 4.6 times, p˂0.001), a decrease of H₂S contents, activity of H₂S-producing enzymes (CSE, CBS and CAT / 3-MST), activity of TRR in the kidney by 33.2–58.1 % (p˂0.001) and an increase of H₂S utilization rates by 79.4 % (p˂0.001) compared with control group. The use of metformin in ST-diabetes reveals hypoglycemic activity (glucose level decreases by 25.2 %, p˂0,001, compared with untreated animals), reduces H₂S deficiency in the kidneys (H₂S level increases by 27.9 %, p˂0.001), increases the activity of H₂S-producing enzymes and TRR (by 15.2–60.0 %, p˂0.05), and also reduces the rate of H₂S utilization (by 32.7 %, p˂0.001). The introduction of the donor H₂S – NaHS potentiates the hypoglycemic activity of metformin and its ability to correct H₂S exchange in the kidneys while the introduction of the inhibitor of H₂S synthesis – PPG reveals the opposite effect in ST-diabetes.

Conclusion. Metformin exhibits hypoglycemic activity and corrects impaired H₂S metabolism in the kidneys in ST-diabetes. The use of NaHS enhanced the hypoglycemic activity of metformin and potentiated its effect on the renal H₂S system in the kidneys while the use of PPG reduced the ability of metformin to correct hyperglycemia and renal H₂S metabolism in the kidneys.

FEATURES OF LIPOPEROXIDATION PROCESSES IN THE PROSTATE AT CHRONIC ALCOHOL INTOXICATION

M. S. Hnatjuk — 2023-02-28

Introduction. Chronic alcohol intoxication leads to damage to almost all organs and systems, the degree of structural and functional disorders of which in this pathology is different and depends on the duration and severity of intoxication.

The aim of the study – to learn the features of lipoperoxidation processes in the prostate gland during chronic alcohol intoxication.

Research Methods. Biochemical and morphometric methods investigated the structures of the prostate of 60 laboratory adult white male rats, which were divided into 2 groups. Group 1 included 30 intact experimental animals, group 2 included 30 rats that were at chronic alcohol intoxication. A month after the start of the experiment, the experimental animals were euthanized by bloodletting under thiopental anesthesia. The content of diene conjugates, active products of thiobarbituric acid, was determined in prostate homogenates. Correlation analysis was carried out between biochemical and studied histostereometric parameters with determination of correlation coefficient (r). Quantitative indicators were processed statistically.

Results and Discussion. It was established that in conditions of chronic ethanol intoxication, the processes of lipoperoxidation in the tissues of the prostate significantly increase. The level of diene conjugates increased by 2.3 times (p<0.001), the concentration of active products of thiobarbituric acid by 2.4 times (p<0.001). Stromal-parenchymatous relation in chronic ethanol intoxication in the studied organ increased by 1.57 times, nuclear-cytoplasmic ratios in endotheliocytes – by 7.6 %, epitheliocytes of glands – by 9.6 % (р<0.001), the relative volume of damaged endotheliocytes – 11.66 times, gland epitheliocytes – 19.7 times (p<0.001). Strong positive correlations between biochemical and morphometric indicators were revealed.

Conclusions. Chronic alcohol intoxication leads to a marked increase in peroxide oxidation of lipids in prostate tissues and damage to the endotheliocytes of arteries and epithelial cells of glands, infiltrative and sclerotic processes. The degree of lipoperoxydation disorders in simulated pathology correlates with the severity of changes in structural homeostasis, the relative volumes of damaged endotheliocytes, glandular epitheliocytes, and stromal-parenchymal relations in the organ under study.

EFFICIENCY OF ENDOTHELIAL DYSFUNCTION CORRECTION OF VESSELS IN THE PERI- AND POSTOPERATIVE PERIOD IN PATIENTS WITH ACUTE CORONARY SYNDROME ON THE BACKGROUND OF TYPE 2 DIABETES

M. I. Shved — 2023-02-28

Introduction. The high level of morbidity and mortality of patients with myocardial infarction and type 2 diabetes and the limitation of the possibility of using highly effective early invasive coronary interventions in this comorbid condition determines the tactics of active study of the role of mutually aggravating pathogenetic factors, including vascular endothelial dysfunction, which contributes to energetic metabolism disorders and ischemic damage of cardiomyocytes.

The aim of the study – to analyze the effectiveness of the correction of endothelial dysfunction of combined interventional and drug treatment in patients with ST elevation acute coronary syndrome against the background of type 2 diabetes with a high risk of developing cardial complications at the inpatient stage of treatment.

Research Methods. 56 patients with acute coronary syndrome in combination with type 2 diabetes were included in the study. They were divided into 2 groups. 32 patients of the main research group, who in addition to standard treatment with acute coronary syndrome received 2.0 g of L-carnitine intravenously in the form of a solution for infusions of 100 ml once per day due to 7 days. One infusion of the drug was performed before interventional intervention. The control group included 24 patients who also underwent percutaneous coronary intervention and standard medical treatment. To compensate for type 2 diabetes, patients of both groups were additionally prescribed dapagliflozin (Forxiga) 10 mg/day. In addition to general clinical, instrumental and laboratory methods, the functional state of the vascular endothelium was determined by the concentration of endothelin-1 (ET-1) in blood plasma and the content of stable NO metabolites.

Results and Discussion. In the initial state, severe microcirculation and endothelial function disorders and their additional deterioration in the peri- and postoperative periods after percutaneous coronary intervention, accompanied by the development of reperfusion syndrome with varying duration and severity of left ventricular failure and rhythm and conduction disturbances, were found in patients with acute coronary syndrome in combination with type 2 diabetes. Under the influence of standard medical treatment, the presence of treatment-resistant supraventricular and ventricular extrasystoles was noted in patients of the control group. At the same time, in patients with ST elevation acute coronary syndrome in combination with type 2 diabetes of the research group, under the influence of complex drug treatment including L-carnitine and dapagliflozin, already on the second day of observation, a significant improvement of the endothelial function of the vessels parameters and a decrease in the frequency of rhythm and conduction disturbances were noted. And after the end of the treatment course in comorbid patients of the research group, a complete recovery of endothelial function and a decrease in the frequency of rhythm and conduction disturbances by 59.4 % and manifestations of acute heart failure by 42.6 % were noted.

Conclusions. Violation of endothelial function in patients with acute coronary syndrome on the background of type 2 diabetes is advisable to be corrected by metabolic therapy with L-carnitine and the sglt2 inhibitor dapagliflozin in the preoperative period of percutaneous coronary intervention to prevent manifestations of reperfusion syndrome and to continue it in the postoperative period to stabilize the clinical condition of patients and prevent complications.

PATHOGENETIC ROLE OF SURFACTANT PROTEIN B IN THE FORMATION OF PULMONARY PATHOLOGY IN ANIMALS WITH STREPTOZOTOCIN-INDUCED DIABETES

L. M. Zaiats — 2023-02-28

Introduction. Nowadays, most researchers consider the lungs to be one of the main targets of diabetes mellitus. According to the literature data, pulmonary surfactants, particularly surfactant protein B play the leading role in the pathogenesis of respiratory diseases.

The aim of the study – to establish the pathogenetic role of surfactant protein B (SP-B) in the formation of lung pathology in animals with streptozotocin-induced diabetes.

Research Methods. The experiments were performed on 88 white male Wistar rats weighing 170–210 g. Animals were divided into three groups: 1 – intact (n=10); 2 – control (n=40); 3 – experimental (n=38) with a model of diabetes mellitus, which was reproduced by intraperitoneal injection of streptozotocin by “Sigma” company (USA), diluted in 0.1 M citrate buffer with a pH of 4.5, at a rate of 60 mg/kg body weight. The control group of animals received an intraperitoneal injection with an equivalent dose of 0.1 M citrate buffer solution with a pH of 4.5. SP-B content in blood serum was determined by Rat enzyme-linked immunosorbent assay (ELISA) kit (Elabscience, the USA) 14, 28, 42, and 70 days after streptozotocin injection.

Results and Discussion. Biochemical studies of blood serum showed an increase in SP-B levels in animals with streptozotocin-induced diabetes at all stages of the research. In particular, serum SP-B levels increased by 8.5 % in 14 days, 37.0 % in 28 days, 54.2 % in 42 days and 74.5 % in 70 days compared to the control group of animals.

Conclusions. During the entire study period, streptozotocin-induced diabetes is accompanied by an increase of surfactant protein B levels in the blood serum which plays a key role in the pathogenesis of lung injury in this pathology.

THE CORRECTIVE EFFECT OF PEPTIDES ON THE PRO- AND ANTIOXIDANT SYSTEM INDICATORS' CHANGES IN RATS OF DIFFERENT AGES AFFECTED BY HEAVY METALS AND ROUNDUP

Ye. B. Dmukhalska — 2023-02-28

Introduction. Heavy metals and organophosphorus compounds used in agriculture cause diseases of the liver and other organs, which contributes to the formation of reactive oxygen species (ROS), which can induce lipid peroxidation and inhibit the antioxidant system. The basis of the action of heavy metals is the blocking of functionally active groups of structural proteins, enzyme proteins, the blocking of sulfhydryl (thiol, SH) groups is of greatest importance. Under the action of heavy metals, most proteins lose their physicochemical and biological properties, which leads to disruption of protein and other metabolism.

The aim of the study – to investigate the corrective effect of peptides on the state of pro- and antioxidant systems in rats of various ages affected by lead acetate, cuprum sulfate, and glyphosate (in the form of roundup herbicide).

Research Methods. Experiments were conducted on laboratory non-linear white male rats of 3 age periods (sexually immature, sexually mature and old), which were administered intragastrically for 30 days with aqueous solutions of lead acetate, copper sulfate and glyphosate. For the purpose of correction, on the 21st day, 6 hours after the introduction of toxicants, peptides were administered for 10 days. Glutathione peroxidase, glutathione reductase, catalase, superoxide dismutase activity and the content of SH-groups, ROS, TBA-active products (TBA-AP) and diene conjugates (DC) were determined in blood serum and liver homogenate of affected and corrected animals by the spectrophotometric method.

Results and Discussion. Heavy metals and organophosphorus compounds caused the formation of ROS, such as superoxide ions, hydrogen peroxide and hydroxyl radicals. With the combined action of lead acetate, copper sulfate, and glyphosate, the processes of free radical oxidation of lipids and the generation of ROS in rats were activated with age, which was evidenced by the increase in the content of DC, TBA-AP, superoxide anion radical, and hydroxyl radical. As our studies showed, the introduction of toxicants led to a decrease in glutathione peroxidase, glutathione reductase, catalase, superoxide dismutase activity, the level of SH-groups in blood serum and liver homogenate of affected animals. The use of peptides as correction factors contributed to a decrease towards the norm in the content of ROS and products of lipid peroxidation and normalization of the activity of enzymes of the antioxidant system, which obviously indicates the antioxidant and chelating properties of peptides.

Conclusion. Exposure of rats to lead acetate, copper sulfate and glyphosate at a dose of 1/20 LD50 leads to an increase in the content of TBA-AP, DC, ROS and a decrease in the activity of enzymes of the antioxidant system in blood serum and liver homogenate. Administration of peptides as corrective factors to animals of various ages with toxic liver damage increases glutathione peroxidase, glutathione reductase, catalase, and superoxide dismutase activity toward normal levels and reduces the content of lipid free radical oxidation products and ROS.

HORMONAL PROFILE IN WOMEN WITH REPRODUCTIVE FAILS AND METABOLIC DISORDERS

S. V. Khmil — 2023-02-28

Introduction. Only the full interaction of the hypothalamic-pituitary-gonadal axis provides the normal functioning of the reproductive system. Hormonal regulation disorders are observed in women of reproductive age with polycystic ovary syndrome (PCOS). Infertility and reproductive losses are characteristic of women with PCOS. Key links of pathogenesis, which are directly related to the occurrence of hormonal imbalance, are also important for understanding right treatment approaches. These are the current issues for further research.

The aim of the study – to determine the levels of sex hormones in women with infertility against the background of PCOS and its correlation with the body mass index.

Materials and Methods. The clinical data analysis of patients’ cards was performed and the level of hormones of the reproductive system was determined. Patients were divided into two groups: with normal body weight and with obesity. In this case, 4 subgroups were identified in the group of studied groups based on the BMI.

Results and Discussion. The analysis of associations between the levels of sex hormones and BMI showed that the LH/FSH ratio and the concentration of FSH, prolactin, estradiol and testosterone depended on body mass. It was established that BMI has a weak inverse relationship with the level of FSH, as well as a direct relationship with the ratio of LH/FSH and the concentration of LH, prolactin, estradiol and testosterone.

Conclusions. Women with infertility on the background of PCOS are characterized by hormonal imbalance (increased levels of anti-Müllerian and luteinizing hormones, estradiol and testosterone, decreased level of FSH).

DEVELOPMENT OF OXIDATIVE PROCESSES IN THE HEART OF RATS DURING THE DEVELOPMENT OF IMMOBILIZATION STRESS

O. V. Denefil — 2023-02-28

Introduction. In recent months, due to the war on the territory of Ukraine, the number of people who spent a long time in a sedentary position in a stressful state has increased significantly. The leading link in the development of any pathology remains the development of oxidative stress, which affects the development of disorders of the cardiovascular system.

The aim of the study – to evaluate the development of oxidative stress in the homogenate of the rat heart during the development of immobilization stress.

Research Methods. Experiments were performed on 60 outbred male rats weighing 180–210 grams. Animals were divided into 5 groups – control, stress 1 day, stress 2 days, stress 3 days, stress 4 days. Stress was modeled by one-hour immobilization of animals lying on the back. Animals in each group were investigated in control, 1 day after the last immobilization. Under thiopental-sodium anesthesia, heart sampling was performed, in the homogenate of which diene conjugates (DC), TBA-active products (TBA-ap), superoxide dismutase (SOD) and catalase activity (Cat) were determined.

Results and Discussion. In male rats, one day after a single episode of stress, compared to the control, an increase in the concentration of DC by 41.4 % (p<0.001) and TBA-ap – by 21.1 % (p<0.01) and an increase in SOD activity by 38.5 % (p<0.001) and catalase activity by 68.5 % (p<0.001) was noted. There was an increase in DC and TBA-ap after 2 and 3 days of stress, but the indicators decreased after 4 immobilizations. There was an increase in SOD and Cat after 2 days of stress, but the indicators decreased after 3 days, and after 4 days, compared to the previous term, it increased.

Conclusion. The development of oxidative stress in the heart of rats under immobilization stress depends on the number of stressful episodes. With three one-hour immobilization, distress develops in male rats, with one and two immobilization – eustress.

THE EFFECT OF L-ARGININE AND AMINOGUANIDINE ON THE DEVELOPMENT OF OXIDATIVE STRESS IN THE LUNGS IN EXPERIMENTAL ANTIPHOSPHOLIPID SYNDROME

N. Ya. Mekhno — 2023-02-28

Introduction. Antiphospholipid syndrome is an autoimmune disease accompanied by the formation of antiphospholipid antibodies which entering into blood coagulation reactions, cause thrombosis of veins and arteries, as well as contribute to miscarriage. At the same time, the processes of free radical oxidation are activated, which is a consequence of ischemia, the antioxidant system is depleted, which deepens lung damage.

The aim of the study – to investigate the state of the prooxidant-antioxidant system in the lung tissue with experimental APS and when using modulators of nitric oxide synthesis (L-arginine and aminoguanidine).

Research Methods. Modeling of the antiphospholipid syndrome was performed on female BALB/c mice. The level of oxidation products and the activity of antioxidant enzymes were determined under APS and with the use of L-arginine and aminoguanidine in the lung tissue.

Results and Discussion. Under conditions of experimental APS in the lungs of mice, an increase in the content of lipid hydroperoxides and TBA-active products, inhibition of the activity of superoxide dismutase and catalase, depletion pool of the reduced glutathione, and impaired function of respiratory chain enzymes – succinate dehydrogenase and cytochrome oxidase. Positive dynamics was noted in restoring the activity of the antioxidant system and blocking the destructive action of free radicals on the lung tissue with the introduction of L-arginine and aminoguanidine and their combination.

Conclusions. Thrombosis and ischemia that occur during APS disrupt the balance of the prooxidant-antioxidant system, increasing the formation of reactive oxygen species and suppressing protective functions. The combined use of L-arginine and aminoguanidine in antiphospholipid syndrome helps to reduce the manifestations of oxidative stress in the lungs, in particular, to reduce the level of lipid peroxidation, restore the activity and content of the components of the antioxidant system, which is accompanied by the restoration of the activity of respiratory chain enzymes.

INFLUENCE OF COMBINED ACTION OF ZINC OXIDE NANOPARTICLES AND HERBICIDE GLYPHOSATE ON THE STATE OF HUMORAL LINK IMMUNE SYSTEM

Ya. Yu. Petrenko — 2023-02-28

Introduction. Nanotechnology today is the most promising direction in the development of world science. Nanomaterials have caused a step forward in many industries and are used in our overall life. Due to their unique properties, nanoparticles are widely used in scientific research, industry and medicine. With everyday increase in the pace of use of nanomaterials, less attention is paid to the possible negative effect of nanoparticles on the health of people and environmental safety as a whole. The capability of nanoparticles to increase the transport of chemicals and drugs into cells and through the body barriers provides the possibility of the potentiating of chemical contaminants toxicity in case of their simultaneous intake into the body.

The aim of the study – to investigate the ability of zinc oxide nanoparticles and chemical toxicant herbicide glyphosate to cause changes in the state of humoral immune system

Research Methods. Suspension of ZnO nanoparticles (0.5 ml) at a dose of 100 mg/kg and glyphosate (in the form of herbicide roundup) at a dose of 250 mg/kg of body weight were administered intragastrically to male rats for 14 days. The toxicants were administered simultaneously and separately. Circulating immune complexes and classes of immunoglobulins А, М, G were measured in rat blood plasma.

Results and Discussion. It was established that under the influence of zinc oxide nanoparticles, the studied indicators did not reliably differ from the control indicators. The administration of glyphosate to rats caused significant changes of indices immunoglobulins G and circulating immune complexes. However, the maximal changes of the parameters were evidenced in the group of animals that were co-administered with zinc oxide nanoparticles and glyphosate. In this case, activities of circulating immune complexes and immunoglobulins blood significantly changed compared to the similar indicators in the group of animals, which were administered with the chemical toxicant only.

Conclusion. Zinc oxide nanoparticles enhance the negative toxic effects of herbicide glyphosate. Carbon nanotubes enhance the negative toxic effects of tetrachloromethane.

DEVELOPMENT OF OXIDATIVE STRESS AT ADRENALINE DAMAGE OF HEART IN MALE RATS WITH CASTRATION AND HYPODYNAMY

R. B. Druziuk — 2023-02-28

Introduction. In recent months, due to the war in Ukraine, the number of people who are in a sedentary position and stressed for a long time has increased. The leading link in the development of any pathology is the development of oxidative stress, which affects the development of disorders of the cardiovascular system.

The aim of the study – to evaluate the effect of castration and hypodynamia on the development of oxidative processes in rats with epinephrine damage of heart (EDH).

Research Methods. Experiments were performed on 120 outbred male Wistar rats. All animals were divided into two series: 1 – control, 2 – castration and stress. Castration was performed under sodium thiopental anesthesia according to the method of Ya. D. Kirshenblatt. Hypodynamic stress was caused from 1.5 to 3 months by keeping them in cages with a limitation of the living space twice for 1.5 months. To reproduce EDH, rats were injected once intraperitoneally with a 0.18 % solution of adrenaline hydrotartrate at the rate of 0.5 mg/kg of weight (Pharmaceutical company "Darnytsia", Ukraine). The animals in each group were examined in control, after 1, 3, 7, 14 and 28 days. Under thiopental-sodium anesthesia, a blood sample was taken, where diene and triene conjugates (DC, TC), Schiff's bases (SB), TBA-active products (TBA-ap), superoxide dismutase (SOD), catalase activity (Cat), ceruloplasmin (CP).

Results and Discussion. In animals that underwent castration and stress, lower values of DC, TC, SB, SOD and CP, but higher TBA-ap and Cat. In rats of the 1st series, with the development of EDH, a greater increase in DC, TC, TBA-ap was noted up to 14 days, and SB decreased after 7 days, with the greatest increase in antioxidants after 7–14 days. In the 2nd series of rats, the maximum values of DC and TC were after 14 days, TBA-ap – after 1 day, SB – after 7 days, with the greatest increase of SOD after 14 days, and Cat and CP – after 1 day.

Conclusion. The combination of castration and stress in male rats causes a decrease in the products of lipid peroxide oxidation and the activity of superoxide dismutase and catalase and the content of ceruloplasmin. The development of EDH caused an increase in the products of lipid peroxidation, which was more pronounced in the control series of animals. Antioxidant activity increased maximally after 14 days in the control series of rats, and after 1 day in animals that underwent castration and stress.

THE ROLE OF NITROGEN (II) OXIDE IN THE MECHANISMS OF THE EXPERIMENTAL HEPATOPULMONARY SYNDROME DEVELOPMENT

I. Ya. Krynytska — 2023-02-28

Introduction. Liver cirrhosis is often accompanied by complications of the pulmonary system, which significantly reduces the life expectancy of such patients. Pulmonary complications of liver cirrhosis include hydrothorax, portopulmonary hypertension, and hepatopulmonary syndrome (HPS), the prevalence of which varies from 5 to 30 %. Nitrogen (II) oxide (NO) is involved in most metabolic processes occurring in the liver. Despite a large number of studies, today there are no clear ideas about the effect of NO and its metabolites on the biochemical mechanisms of the hepatopulmonary syndrome development.

The aim of the study – to investigate changes in the content of nitrogen (II) oxide metabolites in the blood serum and bronchoalveolar lavage of rats with simulated HPS and substantiate the role of NO in the mechanisms of lung damage in experimental HPS.

Research Methods. The experiments were carried out on 56 outbred male rats weighing 180–220 g. The first experimental model of HPS was created by applying a double ligature to the common biliary duct and then cutting it with a scalpel. The second experimental model of HPS was created by 8-week intragastric administration of an oily solution of carbon tetrachloride. Blood serum and bronchoalveolar lavage (BAL) were subjects for investigation. The content of NO metabolites was quantified by determining their sum, including nitrite ions previously present in the sample (NO₂^–) and nitrate ions reduced to nitrite (NO₃^–).

Results and Discussion. The total content of NO metabolites in the blood serum of rats on the 31st day after ligation of the common bile duct significantly increased by 3.9 times (p₁<0.001) compared with the control group No. 1. In rats with carbon tetrachloride-induced cirrhosis, the total content of NO metabolites in blood serum increased by 3.1 times (p₁<0.001). Changes in NO production occurred unidirectionally in the direction of deepening oxidative stress both in blood serum and in BAL.

Conclusions. When modeling HPS, nitroxidergic processes are activated due to a possible increase in the content of NO metabolites in the blood serum and BAL. When comparing the absolute values of the content of NO metabolites in the blood serum and BAL, a synchronous development of nitroxidergic processes at the systemic and local levels was revealed with a predominance of nitrogen (II) oxide synthesis in the lungs.

METHODS OF ENZYMATIC HYDROLYSIS FOR BLOOD SAMPLE PREPARATION FOR LABORATORY DIAGNOSTICS OF POISONING WITH GAMMA-AMINO BUTYRIC ACID DERIVATIVES

N. V. Horlachuk — 2023-02-28

Introduction. New potentially dangerous psychoactive substances, which are not included in the list of narcotic drugs and psychotropic substances, the circulation of which is prohibited or restricted in Ukraine, include ten groups of substances of different structures, among which a group of GABA-A and GABA-B receptor agonists and their derivatives can be distinguished. Abuse of new dangerous psychoactive substances, including GABA derivatives, has been recorded in 94 countries of the world, including Ukraine. In connection with the recent increase in the number of people addicted to these drugs, as well as the number of acute and even fatal poisonings, the issue of chemical and toxicological research, which includes joint detection and quantitative determination of analyzed substances in biological bodies, is becoming more and more relevant projects.

The aim of the study – to develop methods for the isolation and determination of GABA derivatives (phenibut and baclofen) in biological fluids (blood).

Research Methods. The research was conducted using the substance phenibut - (±)-4-amino-3-phenylbutanoic acid hydrochloride and baclofen tablets. Enzymes were used for hydrolysis: papain, chymotrypsin, chymopsin and hyaluronidase, trilon B substance, cysteine substance

Results and Discussion. A significant difficulty in conducting research on biological objects for GABA derivatives (phenibut, pregabalin, gabapentin, baclofen, etc.) is that these substances are able to produce internal salt in the environment of biological fluids, which significantly complicates conducting laboratory studies on them. In our work, we used the method of extraction freezing, determining the selective conditions, pH 2 with acetonitrile at a temperature of -18 ... -20 C.

Conclusion. In the course of the conducted research, it was shown that the method of enzymatic hydrolysis by proteases of low specificity can be recommended for blood sample preparation in the diagnosis of non-medicinal use of drugs from the group of GABA derivatives.

ANALYTICAL CONTROL OF CEREAL CROP PROTECTION SYSTEM PESTICIDE RESIDUES IN THE WATER AND AIR

S. T. Omelchuk — 2023-02-28

Introduction. Chromatographic analysis to develop conditions for determining 9 active substances (prosulfuron and pinoxaden herbicides, thiamethoxam and lambda-cyhalothrin insecticides, azoxystrobin, benzovindiflupyr, cyproconazole, propiconazole fungicides, trinexapac-ethyl plant growth regulator) of the cereal crop protection system in their combined presence was performed using analytical standards from which the initial standard solutions of each of the test compounds were prepared in acetonitrile. Chromatographic analysis of each calibration solution of the mixture was performed 3 times to construct the graphs of dependence between the area of the compound chromatographic peak and its concentration. Quantitative determination of the test compound was performed by the method of absolute calibration (external standard method) and the obtained results were processed by statistical methods.

The aim of the study – development and improvement of analytical control of residual amounts of pesticides in water and air when it used as a part of grain protection system.

Research Methods. The findings allowed us to calculate the retention factor (k) and to construct dependences of relative retention of test compounds on columns C18 and CN on the content of acetonitrile in the mobile phase acetonitrile + 0.1 % aqueous solution of orthophosphoric acid and to continue research on compounds separation in its combined presence as more effective using column 250/4.6 Nucleosil 100-5 C18 and a mixture of acetonitrile + 0.1 % aqueous solution of orthophosphoric acid. According to the results of studying the absorption spectra, in order to improve the chromatograms of water samples and, more particular, air, the wavelength of the UV detector was optimized, which gave a positive result.

Results and Discussion. Conditions for water and air samples preparation for further chromatographic determination of test compounds and optimal conditions for extraction and chromatographic determination of different chemical class pesticides used in the cereal crops protection system. These conditions allow to control pesticides shared content in the water and air sample with limits of detection – 0.001 mg/dm³ and 0.05 mg/m³, respectively i.e., will allow controlling the approved hygienic standards of these compounds in the water and working zone air.

Conclusion. Developed conditions for determining 9 cereal crop protection system pesticides by high-performance liquid chromatography method in its shared presence in water and air samples, can significantly speed up the analysis, reduce costs for analysis performance and improve pesticides monitoring in the environment.

CHANGES IN THE CONTENT OF MIDDLE WEIGHT MOLECULES IN FEMALE RATS WITH OVARIAN PATHOLOGY AND THEIR CORRECTION BY INOSITOL

N. Y. Terletska — 2023-02-28

Introduction. The problem of infertility is still relevant today. The main reasons for its development are: age; stress, sedentary lifestyle, exposure to heavy metal salts. Inositol preparations (In) improve a woman's reproductive function.

The aim of the study – to assess the development of endogenous intoxication in female rats with ovarian pathology and to carry out correction with In preparations.

Research Methods. Experiments were performed on 144 white female Wistar rats, which were divided into 4 series: 1 – control, 3.5-4-month-old rats, 2 – rats of 7.5–8 months, 3 – rats that were intragastrically injected with lead acetate solution at a dose of 0.05 mg/kg for 15 days , 4 – hypodynamic stress. There were 3 groups in each series: A – control (C), B – In, B – FT 500 plus (FT). In and FT was administered intragastrically for 15 days at a dose of 400 mg/kg of In. Middle weight molecules (MWM) were determined in blood serum at λ 280 nm, 260, 254, and 238 nm.

Results and Discussion. In the control, MWM₂₃₈ were highest in 3A. In 2A and 4A, the values were lower than K by 35.7 % and 67.9 %. MWM₂₅₄ increased in 2A and 3A, compared to 1A, by 2.4 times and 3.0 times. MWM₂₆₀ were greater than the indicators of 1A only in the 2nd series by 44.4 %. MWM₂₈₀ were the smallest in the K series of rats. Yin in rats of the 1st series caused a decrease in MWM₂₃₈ and MWM₂₈₀, and an increase in MWM₂₅₄ and MWM₂₆₀. FT resulted in a decrease in MWM₂₃₈ but an increase in MWM₂₅₄ and MWM₂₈₀. In series 2, In contributed to the reduction of MWM₂₅₄ and MWM₂₈₀. FT resulted in a decrease in all MWM. In series 3, Ying and FT contributed to the decrease of MWM₂₃₈ and MWM₂₈₀, and the growth of MWM₂₆₀. In series 4, In caused a decrease in MWM₂₈₀ but an increase in MWM₂₅₄. FT resulted in increased MSM₂₃₈, MWM₂₅₄ and MWM₂₈₀.

Conclusion. In preparations at a dose of 400 mg/kg should not be used in an experiment for healthy young female rats. In 7.5–8 month old animals, FT is optimal for reducing endogenous intoxication. In animals exposed to nanodoses of lead acetate, the effectiveness of using inositol and FT is the same. The use of inositol preparations is not effective in animals that have been in hypodynamia and stress for a long time.

EVALUATION OF THE HORMONAL BACKGROUND OF WOMEN WITH ENDOMETRIOSIS-ASSOCIATED INFERTILITY

S. V. Khmil — 2023-02-28

Introduction. Endometriosis is an estrogen-dependent chronic inflammatory disease that causes endometriosis-associated infertility in approximately 30–50 % of women with a confirmed case.

The aim of the study – to analyze the reproductive hormonal panel of patients with endometriosis-associated infertility to choose the optimal ovulation stimulation protocol in the programs of assisted reproductive technologies.

Research Methods. In accordance with the purpose, a comparative retrospective analysis of the results of the hormonal panel was carried out in 116 women with infertility on the background of endometriosis with long and short ovulation stimulation protocols and a control group in the programs of assisted reproductive technologies. Patients were divided into 3 groups. Group 1 consisted of 44 women with endometriosis-associated infertility who underwent controlled ovarian stimulation according to a long protocol with a-GnRH. Group 2 consisted of 42 patients with endometriosis-associated infertility who underwent controlled ovarian stimulation according to a short protocol with anti-GnRH. Comparison group 3 (control) consisted of 30 women with tubal factor infertility. The level of anti-Mullerian hormones (AMH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), prolactin, estradiol E2, and total testosterone in blood serum was determined by the enzyme-linked immunosorbent assay (ELISA) method in the laboratory of the Medical Center “Clinic of Professor Stefan Khmil” (accreditation No. 268604/2019). Indicators of the listed hormones were determined on certain days of the menstrual cycle. The level of total testosterone, follicle-stimulating, luteinizing hormone, estradiol was determined on the day 2–3 of the cycle, the level of progesterone was determined on the day 21–22 of the cycle, and on the day of the trigger injection the level of estradiol and progesterone was determined. The level of all the listed hormones was determined by the immunoenzyme analyzer “StatFax” and the test system “Diagnostic Systems Laboratories, Inc.” (USA).

Results and Discussion. A certain regularity of the hormonal panel of women with endometriosis-associated infertility was established, in particular, an increased level of FSH, an increased level of LH, a decreased ratio of LH to FSH, decreased anti-Mullerian hormone, which correlated with the degree of prevalence of the process, progesterone and androgen deficiency. However, the hyperestrogenic state inherent in endometriosis has been found to directly contribute to lesion growth through its proliferative and inflammatory effects on endometrial tissue.

Conclusions. Endometriosis-associated infertility is caused by a malfunction of the hypothalamic-pituitary-ovarian system, which requires more detailed further study in order to develop new technologies and treatment protocols. Undoubtedly, the best method of treating endometriosis associated infertility is the use of assisted reproductive technologies, however, patient management tactics and the stimulation protocol must be selected individually for each individual case of the diagnosis, depending on the reproductive potential (ovulatory reserve, hormonal profile) and the woman's history.