Thiopental Anesthesia Research Articles

Changes in morphometric indicators of liver blood vessels under conditions of long-term ethanol exposure

Hyperdynamic splanchnic blood circulation is an important component of portal hypertension syndrome, which develops under conditions of chronic alcoholic hepatitis. Angiogenesis caused by alcohol consumption contributes to the development of splanchnic hyperemia and the development of portal-systemic collaterals. The purpose of the study is to study the morphometric indicators of the vascular bed of the liver of rats under the conditions of chronic alcoholic hepatitis modeling. Experiments were performed on 30 white, sexually mature male Wistar rats weighing 180–220 g. The animals were divided into 2 groups: I (control group) contained 6 rats; II – 24 rats that modeled chronic alcoholic hepatitis by the method of forced intermittent alcoholization for 5 days, with a repeat after two days by intraperitoneal injection of a 16.5% ethanol solution in a 5.0% glucose solution, at the rate of 4 ml/kg of body weight. Animals were removed from the experiment on days 35, 42, 49, and 56 by taking blood from the right ventricle of the heart under thiopental anesthesia. On the 35th day of simulation of chronic alcoholic hepatitis, the inner diameter of central veins, lobular arterioles and interlobular veins increased, and interlobular arteries and lobular venules decreased compared to the control. On the 42nd day of simulation of chronic alcoholic hepatitis, the internal diameter of central veins, lobular arterioles and interlobular veins increased, and interlobular arteries decreased compared to the control. On the 49th day of simulation of chronic alcoholic hepatitis, the inner diameter of the central veins, lobular arterioles, and interlobular veins increased, and the interlobular arteries decreased compared to the control. On the 56th day of simulation of chronic alcoholic hepatitis, the inner diameter of the central veins and lobular arterioles increased, and the interlobular arteries and lobular venules decreased compared to the control. Keywords: arteries, veins, arterioles, venules, sinusoidal capillaries.

OXIDATIVE MODIFICATION OF PROTEINS IN COLORECTAL CANCER UNDER THE INFLUENCE OF 5-FLUOROURACIL AND MOLECULAR HYDROGEN

The aim of the study – to investigate the effect of water saturated with molecular hydrogenand 5-fluorouracil on the content of carbonyl groups of oxidatively modified proteins inthe blood serum of white rats with colorectal cancer.Material and Methods. The experiments were conducted on 50 male Wistar rats. Theanimals were modeled with colorectal cancer (CRC) by subcutaneous injection of1,2-dimethylhydrazine (DMH) at a dose of 7.2 mg/kg body weight once a week for 30weeks. 5-Fluorouracil was administered intraperitoneally for 4 days at 12 mg/kg andfor another 4 days every other day at 6 mg/kg. Animals consumed water enriched withmolecular hydrogen at a concentration of 0.6 ppm ad libitum. Rats were euthanizedunder thiopental anesthesia. Blood serum was used for the study, in which the content ofcarbonyl groups was determined by the colorimetric method. Statistical data processingwas performed using the SPSS-22 software package.Results. It was found that the modeling of colorectal cancer led to a 1.77-fold increase inthe content of carbonyl groups in the blood serum of rats compared to intact animals. Thecontent of carbonyl groups in the blood serum of rats with CRC treated with chemotherapyby administration of 5-fluorouracil increased 1.93 times compared to the intact groupand 1.1 times compared to animals with CRC. Consumption of water saturated withmolecular hydrogen 30 days after modeling CRC and administration of 5-fluorouracilto white rats led to a 1.18-fold decrease in the content of carbonyl groups in their bloodserum compared to animals with CRC who were administered 5-fluorouracil but did notconsume water saturated with molecular hydrogen.Conclusions. The use of molecular hydrogen saturated water is an effective methodof reducing oxidative modification of proteins in rats with colorectal cancer under5-fluorouracil chemotherapy.

Neuroprotective Effects of Bexarotene and Icariin in a Diabetic Rat Model.

Objective Type 2 diabetes mellitus (T2DM), a chronic metabolic disorder affecting over 400 million people globally, is increasingly recognized for its detrimental impact on the central nervous system. T2DM is linked to neurodegenerative diseases like Alzheimer's and vascular dementia. This study investigates the neuroprotective effects of bexarotene and icariin in a T2DM rat model, focusing on brain-derived neurotrophic factor (BDNF), glial fibrillary acidic protein (GFAP), and neurofilament-light chain (NfL) levels. Methods Before the study, rats underwent fasting blood glucose tests, lipid profile assessments, and general health evaluations, followed by a high-fat diet for two weeks and a single streptozotocin dose (35 mg/kg). Rats with fasting blood glucose levels ≥250 mg/dl were classified as diabetes mellitus (DM) and continued on the high-fat diet throughout the experiment. Forty-seven male Wistar Albino rats were divided into six groups: a healthy control group, a DM control group, a DM group treated with bexarotene, a DM group treated with icariin, and two DM groups treated with combinations of low and high doses of bexarotene and icariin. After the 45-day treatment, blood samples were collected under thiopental sodium anesthesia, with HbA1c (glycosylated hemoglobin) and hematological parameters analyzed within eight hours, and serum stored at -80°C for further analysis. The animals were then euthanized, and brain tissues were harvested, frozen, and stored at -80°C until further examination. Brain tissues were analyzed for BDNF, GFAP, and NfL levels using ELISA (enzyme-linked immunosorbent assay). For comparing multiple groups, the Kruskal-Wallis test was applied to nonparametric data, and one-way ANOVA was used for parametric data, followed by Bonferroni's post hoc test for pairwise comparisons. Statistical significance was determined with two-tailed tests at p < 0.05. Results Significant changes in GFAP levels were observed across groups (p < 0.001). The DM control group showed the highest GFAP levels, while treatment groups exhibited reductions. The DM control group also showed the highest BDNF levels, while treatment groups exhibited reductions. The DM control group showed the lowest NfL levels, while treatment groups exhibited increments. Conclusion This study highlights the neuroprotective potential of bexarotene and icariin in a diabetic rat model, evidenced by significant changes in GFAP levels. The lack of significant changes in BDNF and NfL suggests that longer study durations may be necessary to observe these effects. Future research should include extended study periods, larger sample sizes, varied dosages, and comprehensive behavioral assessments to better understand the therapeutic potential of these agents.

Characteristics of HIF-1Α and HSP70 MRNA Expression, Level, and Interleukins in Experimental Chronic Generalized Periodontitis.

Periodontal diseases are a rather complex problem of modern dentistry and do not have only medical but also social significance. The objective of this study is to weigh the effect of a mixture of Thiotriazoline and L-arginine (1:4) on the parameters of the system of endogenous cytoprotection of blood and periodontal illness in rats with experimental chronic generalized periodontitis and substantiate further study of this blend. The study aimed to evaluate the impact of a combination of Thiotriazoline and L-arginine (in a ratio of 1:4) on the parameters of the endogenous blood cytoprotection system and periodontium in rats with experimental chronic generalized periodontitis. A group of outbred rats weighing 190-220 g and sourced from the vivarium of the Institute of Pharmacology and Toxicology of the Academy of Medical Sciences of Ukraine were divided into four groups, each consisting of 10 animals. (1) Intact group, animals that were injected intragastrically with a solution of sodium chloride to chloride 0.9% for 30 days. (2) control, animals with experimental CGP who intragastrically sodium chloride solution 0.9% for 30 days. (3) animals with experimental CGP were injected intramuscularly with Thiotriazoline + L-arginine (1:4) in a dosage of 200 mg/kg (30 days). (4) animals with experimental CGP, for which daily intragastric reference drug Mexidol, in dosage 250 mg/kg (30 days). In this study, we utilized two substances: Thiotriazoline and L-arginine hydrochloride. The combination of Thiotriazoline and L-arginine (in a ratio of 1:4) was prepared at the Department of Pharmaceutical Chemistry of ZSMU. At the conclusion of the experiment, the rats were carefully removed from the study while under thiopental-sodium anesthesia, and administered at a dosage of 40 mg/kg. We have found that the administration of a combined preparation of Thiotriazoline with L-arginine to rats with CGP leads to a significant decrease in the blood concentration of pro-inflammatory cytokines IL-1b and TNF-a by 56.1% and 71%, respectively. The administration of Mexidol at a dosage of 250 mg/kg, as well as the combination of Thiotriazoline and Larginine in a ratio of 1:4 at a dosage of 200 mg/kg, resulted in a significant reduction in gingival pocket depth in animals with CGP. Specifically, the gingival pocket depth was reduced to 6 mm (p < 0.05) with Mexidol and further reduced to 4 mm (p < 0.05) with the combination of Thiotriazoline and L-arginine. Additionally, the animals exhibited minimal bleeding, swelling, and tooth mobility when treated with the combination of Thiotriazoline and L-arginine. The administration of a combination of Thiotriazoline and L-arginine (in a ratio of 1:4) at a dosage of 200 mg/kg to animals with CGP resulted in a noteworthy reduction in the blood concentration of pro-inflammatory cytokines IL-1b and TNF-a. Specifically, there was a significant decrease of 56.1% (p < 0.05) in IL-1b and 71% (p < 0.05) in TNF-a levels. The course administration of a combination of Thiotriazoline and L-arginine (1:4) (200 mg/kg) to animals with CGP led to an increased expression of HSP70 mRNA (p < 0.05) in the periodontium by 8.2 times and HIF-1a mRNA by 8.2 times. 2.8 times (p < 0.05) against the background of an increase in the blood concentration of HSP70 by 95% (p < 0.05). Also, in the periodontium of animals in this group, a decrease in the expression of c-Fos mRNA by 36.7% (p < 0.05) was found compared to the control group.

QUANTITATIVE MORPHOLOGICAL ANALYSIS OF THE FEATURES OF REMODELING OF HEMOMICROCIRCULATORY VESSELS IN THE TESTIS OF EXPERIMENTAL ANIMALS AT CONDITIONS OF ETHANOL INTOXICATION

Ethanol is classifi ed as a vascular poison, long-term intoxication of which negatively aff ects almost all organs and systems body of humans and experimental animals. Morphological changes in testes during ethanol poisoning have not been studied enough. The aim of the study was a quantitative morphological analysis of the features of the remodeling of blood vessels of the hemomicrocirculatory bed of the left and right testes at conditions of ethanol intoxication. The structures of the left and right testis of 30 white male rats, which were divided into 2 groups, were morphologically and morphometrically studied. The 1 group included 15 intact animals, the 2-15 rats, which were injected intragastrically with a 30 % ethanol solution at the rate of 2 ml per 100 g of body weight for 28 days once a day. A month after the start of the experiment, the experimental animals were euthanized by bloodletting under thiopental anesthesia. Histological and semithin sections were made from testes. The outer and inner diameters, wall thickness of arterioles, precapillary arterioles, hemocapillaries, postcapillary venules and venules were determined. Quantitative indicators were processed statistically.It was established that twenty- eight days of ethanol intoxication in laboratory sexually mature white male rats leads to pronounced remodeling of the blood vessels of the hemomicrocirculatory bed of the left and right testes, which is characterized by narrowing of the lumens and thickening of the walls of its arterial (arterioles, precapillary arterioles) and exchange (hemocapillaries) vessels. pronounced expansion and thinning of the walls of postcapillary venules and venules, venous hemoptysis, hypoxia, dystrophic- necrotic changes in cells, stromal structures, infi ltration and sclerosis. The revealed remodeling of blood vessels of the hemomicrocirculatory bed in conditions of ethanol poisoning dominates in the left testicle.

Neuropathological changes in the TASTPM mouse model of Alzheimer’s disease and their relation to hyperexcitability and cortical spreading depolarization

Although Alzheimer’s disease (AD) is characterized by distinct pathological changes, their precise impact on cortical functions are not well understood. Here we used TASTPM mice as an AD model and asked whether the development of neurodegenerative changes has an impact on the extracellular space (ECS) and neuronal excitability, in particular cortical spreading depolarization (CSD) which requires intact neuron and glial functions. We studied wildtype (WT) and TASTPM mice (3, 6, and 12 months old). TASTPM mice showed progressive proliferation of neocortical Amyloid-beta (Aβ) plaques between 3 and 12 months (more deposits in females than in males) and Aβ accumulation in cortical vessels. As plaques proliferated, neuroinflammatory microglial reaction (CD68, CD39 and Galectin-3) and astrogliosis (GFAP) developed progressively. The cortical ECS volume shrank significantly to about half the size of the WT. CSD in both WT and TASTPM mice showed considerable heterogeneity but did not correlate with the histological changes. However, CSDs were easier to elicit in TASTPM than in WT mice at 3 months, and also compared to older TASTPM mice. Moreover, TASTPM mice showed more hyperexcitability manifested as clonic-tonic behavior after sodium thiopental anesthesia. Thus, AD pathology was associated with abnormal hyperexcitability but did not homogenously alter CSD susceptibility.

Submicroscopic changes of type II alveolocytes in the late development of experimental acute pancreatitis

Background. Today, acute pancreatitis is one of the most common and severe visceral disorders in patients worldwide. The aim of the work was to study the dynamics of ultrastructural changes of type II alveolocytes in the long-term periods of development of experimental acute pancreatitis. Material and methods. The experiments were carried out on 54 white Wistar male rats weighing 180–220 g. The animals were divided into three groups: first — intact, second — control, third — experimental with a model of acute pancreatitis, which was reproduced by intraperitoneal administration of a 20% solution of L-arginine at a total dose of 5 g/kg at one-hour interval. The control group of animals was intraperitoneally injected with an equivalent dose of isotonic sodium chloride solution. All research were performed under sodium thiopental anesthesia at the rate of 60 mg/kg body weight. Lung tissue for electron microscopic examination was collected from the lower lobe of the left lung at 3–5 and 7 days. Pieces of lung tissue measuring 1×1×1 mm were fixed in a 2.5% glutaraldehyde solution, followed by additional fixation in a 1% osmium tetroxide solution. After dehydration, the material was poured into Epon-Araldite. Sections with a thickness of 20–50 nm obtained on “Tesla BS-490” ultramicrotome were studied in a PEM-125K electron microscope. Results. It was found that three days after the start of the study, along with dystrophic-destructive changes, type II alveolocytes in a state of increased functional activity were observed. Continuation of the experiment (5 days) leads to the progression of ultrastructural changes in type II alveolocytes are determined, which are most pronounced on the 7 day of the study. Conclusion. Acute experimental pancreatitis is accompanied by marked changes in the ultrastructural structure of type II alveolocytes. The nature and severity of structural changes of type II alveolocytes depends on the duration of the course of arginine-induced acute pancreatitis.&#x0D;

The influence of chronic hyperhomocysteinemia on the structure and immune processes of the spleen in young rats

Homocysteine is a sulfur-containing amino acid that is an intermediate product of methionine metabolism. Entering the body with products of animal origin, methionine undergoes a number of biochemical transformations. Hyperhomocysteinemia has a negative effect on the body, causing damage to all organs and systems and leading to disruption of homeostasis. The study aims to study the characteristics of changes in the structure and immune processes of the spleen of young rats with chronic hyperhomocysteinemia. Experiments were performed on 22 white male rats. The animals were divided into control and experimental groups (11 individuals in each group) during the experiment. Chronic hyperhomocysteinemia was achieved by administering D, L-thiolactone homocysteine hydrochloride to experimental group animals at a dose of 200 mg/kg of body weight intragastrically in a 1 % starch gel solution once a day for eight weeks. After the end of the experimental simulation of chronic hyperhomocysteinemia, the animals were removed from the experiment by anaesthetising by decapitation and using thiopental anaesthesia. Histological preparations were studied using an SEO SСAN light microscope. Experimental modelling of chronic hyperhomocysteinemia in young rats was not accompanied by significant morphological changes in the stroma and red pulp of the spleen. T-cell periarterial sheaths and B-cell nodules in animals from the experimental group underwent changes. B-cell proliferation leads to growth within the white pulp, expansion of the marginal zones, which become more blurred and infiltration of these cells into the red pulp. Evaluation of the red pulp in the spleen of young rats from the experimental group showed a particular expansion in the venous sinuses, which may be a sign of reaction to the influence of homocysteine and slight swelling of these vessels. The number of T-cells in young animals in the periarterial cuffs is slightly reduced, which can be explained by the increasing activity of macrophages. However, T-cells also infiltrated the red pulp. Such penetration of white blood cells into the red pulp is accompanied by the expansion and “washing out” of the marginal zones of lymph nodes. Thus, the increased homocysteine level significantly potentiated proliferation and partially inhibited apoptosis in T-lymphocytes but did not directly affect dormant T-cells.

Ultrastructural organization of alveolar macrophages during late development of experimental acute pancreatitis

Background. Nowadays, acute pancreatitis continues to remain an actual problem both in surgery and in intensive care medicine. The aim of the work was to study the dynamics of ultrastructural changes of alveolar macrophages in the long-term development of experimental acute pancreatitis. Material and methods. The experiments were carried out on 62 white Wistar male rats weighing 180–220 g. The animals were divided into three groups: first — intact, second — control, third — experimental with a model of acute pancreatitis, which was reproduced by intraperitoneal administration of a 20% solution of L-arginine in a total dose of 5 g/kg at one-hour interval. The control group of animals was intraperitoneally injected with an equivalent dose of isotonic sodium chloride solution. All research were performed under sodium thiopental anesthesia at the rate of 60 mg/kg body weight. Lung tissue for electron microscopic examination was collected from the lower lobe of the left lung at 3–5 and 7 days. Pieces of lung tissue measuring 1×1×1 mm were fixed in a 2.5% glutaraldehyde solution, followed by additional fixation in a 1% osmium tetroxide solution. After dehydration, the material was poured into Epon-Araldite. Sections with a thickness of 20–50 nm obtained on “Tesla BS-490” ultramicrotome were studied in a PEM-125K electron microscope. Results. It was found that three days after the start of the study, significant heterogeneity of alveolar macrophages was observed in the alveolar lumen. Next to active phagocytic macrophage elements with dystrophic-destructive changes are determined. Continuation of the experiment (5 days) leads to the progression of submicroscopic changes in alveolar macrophages, which are most pronounced on the7 day of the study. Conclusion. Acute experimental pancreatitis is accompanied by pronounced changes in the ultrastructural structure of alveolar macrophages. The features and severity of structural changes in alveolar macrophages depends on the duration of arginine-induced acute pancreatitis.

Quantitative morphological analysis of age structural changes in prostate of experimental animals with ethanol poisoning

Aim: To find out the age remodeling of the structural components of the prostate gland at alcohol poisoning using quantitative morphological analysis. Materials and Methods: The structure of the prostate gland of 4 white male rats groups were morphologically investigated. The 1 group included 30 control intact animals aged 8 months, the 2-nd group – 30 rats aged 24 months, the 3-rd group – 30 8-month-old animals with ethanol intoxication, and the 4-th group included 30 24-month-old rats with the specified simulated pathology. Ethanol intoxication was modeled by intragastric administration of 30% ethyl alcohol solution at a dose of 20 ml/kg once daily for 28 days. Rats were euthanized by bloodletting under general thiopental anesthesia 28 days after the beginning of the experiment. The area of glands, the height of glandular epithelial cells, the area of their nuclei and cytoplasm, the nuclear-cytoplasmic ratio in these cells and the stromal-parenchymal ratio in the organ were studied using light microscopy and were determined morphometrically. Morphometric parameters were processed statistically. Results: It was established that with age in the intact prostate of laboratory sexually mature white male rats, the area of glands, the height of glandular epitheliocytes, the area of their nuclei and cytoplasm, with the stability of nuclear-cytoplasmic ratios in the epithelial cells of the glands, significantly decreases, and the stromal-parenchymal ratio in the organ under study increases. Long-term ethanol poisoning leads to pronounced structural changes in the prostate, which is characterized by pronounced atrophy of the glandular epithelium, a decrease in the area of the glands, a decrease in the height of epithelial cells, a violation of nuclear-cytoplasmic relations in them, an increase in stromal-parenchymal ratio, and a prominent growth of the muscle-elastic stroma. The revealed structural changes of the studied components of the prostate dominated in 24-month-old experimental animals. Conclusions: Morphological analysis of the prostate gland established that morphometric and morphological changed significantly according to the age and were depend on the ethanol poisoning.

The structural rearrangement of testicular arteries in case of chronic ethanol intoxication

Prolonged poisoning of the body with ethanol damages almost all its organs and systems. Structural changes in testicular arteries under the influence of chronic ethanol intoxication have not been fully studied. Purpose - to analyze the morphometric parameters of the small caliber arteries of the right and left testicles of rats in case of chronic ethanol intoxication. Materials and methods. 62 laboratory sexually mature male rats was performed. The animals were divided into two groups. The Group 1 (control) included 30 rats, the Group 2 (experimental) - 32 animals. The experimental rats, which were conducted to the experiment, were injected intragastrically with a 30% ethanol solution at the rate of 2 ml per 100 g of body weight for 28 days once a day. Animals were euthanized by heart bleeding under thiopental anesthesia. Histological micro-sections were made from the left and right testicles. Morphometrically, the outer and inner diameters of the arteries, media thickness, adventitia, Vogenvoort and Kernogan indices, the height of endotheliocytes, the diameter of their nuclei, the nuclear-cytoplasmic ratio in these cells, and the relative volume of damaged endotheliocytes were determined morphometrically in small-caliber arteries. Statistical processing of digital data was carried out using Excel (Microsoft, USA) and STATISTICA 6.0 (Statsoft, USA) software. Results. It was established that in case of simulated experimental conditions of chronic ethanol intoxication, the investigated morphometric parameters of the arteries of the left and right testicles changed significantly. In case of chronic intoxication with ethanol in rats, pronounced structural rearrangement of testicular vessels, mainly small arteries, characterized by thickening of the blood vessel’s wall, narrowing of vessels’ lumen, rearrangement of endothelial cells, and an increase in the relative volume of the damaged cells of intima, is established, and is dominant in the left testis. Conclusions. In case of chronic intoxication with ethanol the structural changes in testicular tissues are dominant in the left testis. Pronounced remodeling of small caliber testicular arteries causes endothelial dysfunction, deterioration of blood supply to the testicles, hypoxia, dystrophic and necrobiotic tissue changes, infiltrative and sclerotic processes, which are the pathomorphological basis of male infertility. When carrying out experiments with laboratory animals, all bioethical norms and recommendations were observed. No conflict of interests was declared by the authors.

HISTOMORPHOMETRIC ASSESSMENT OF CHANGES THE STRUCTURAL COMPONENTS OF THE BONE IN RATS WITH CHRONIC CHOLESTASIS WITH CORRECTION OF THE COMPLEX FROM HEPATOPROTECTOR AND SORBENT

Problem. The disruption of the gastric duct in cholestasis causes insufficient absorption in the small intestine of macro- and microelements and fat-rich vitamins (A, D, E, K), which are necessary for healthy processes, and remodeling of bone tissue. To reduce pathological changes in bone tissue diseases of the hepatobiliary system in rats statement in cholestasis, need a comprehensive approach [6]. In the core of the preventive complex one can see a hepatoprotector, which, due to its main components (lecithin, quercetin), possesses the liver protecting properties, as well as has anti-inflammatory and bile-diluting effects, and a sorbent. Aim. To conduct a histomorphometric research of the femur in rats with chronic cholestasis and evaluate the effectiveness of the prophylactic complex. Methods. The follow-up study was carried out on male rats, divided into three groups: 1st group – intact, 2nd group – rats with simulated chronic cholestasis, 3rd group – rats, who, in parallel with the development of chronic cholestasis pathology, carried out prevention. Cholestasis in rats was modeled by ligation of the biliary duct under thiopental anesthesia [9]. A hepatoprotector at a dose of 500 mg/kg and a sorbent at a dose of 1 g/kg were administered to group 3 of the animals once over a period of 4 months. Histological analysis and photography of sections of the stegnus cyst were carried out using an additional light microscope and a camera for an Olympus LC30 microscope (Japan). Morphometric studies of rat femur were carried out using the following parameters: volume (%) of the articular epiphyseal cartilage, volume (%) of the cartilage, volume (%) of the cerebrum and the number of osteoblasts per the particular area. The main results. Certain histological changes in the epiphysis of the femoral bone were found in rats with chronic cholestasis, namely, noticeable delamination of the periosteum was noted. In the deep layers of the cartilage, a decrease in the number of isogenic groups was noted, in addition, they contained a smaller number of chondrocytes. Desolation of individual lacunae, a decrease in the number of osteoblasts, and thinning of bone trabeculae were also observed, indicating certain pathological changes in bone tissue. Analysis of the morphometric parameters of the femur in rats with chronic cholestasis revealed a decrease in the specific volume of the cartilage of the articular surfaces by 25.8%, a decrease in the specific volume of the bone marrow and the number of osteoblasts by 14.4% and 39.4%, respectively, and as well as an increase of 33.1% in the specific volume of bone beams in relation to the control group. The introduction of the prophylactic complex contributed to a significant increase in the specific volume of the cartilage of the articular surfaces – by 17.2%, the specific volume of bone marrow by 42.9%, the average number of osteoblasts by 34.9%, and a decrease in the specific volume of bone beams by 31 .7% relative to the group of animals with chronic cholestasis. That is, the use of a prophylactic complex, which consisted of a hepatoprotector and a sorbent, effectively eliminated pathological changes in bone tissue that arose against the background of the development of chronic cholestasis. Conclusions. The formation of chronic cholestasis leads to pathological changes in the cartilaginous tissue of the epiphysis of the femur of the animals under study, and it itself changes the volume of cartilage of the subglobular surfaces and causes destruction of the main components in bone tissue (reduces the volume of bone marrow and the number of osteoblasts at the background of the growth the volume of bone beams). The effectiveness of prophylactic in the elimination of pathological changes in bone tissue at the background of chronic cholestasis has manifested itself as a hepatoprotective, antioxidant, anti-inflammatory and anti-inflammatory hepatoprotector and those combined with a sorbent.

PECULIARITIES OF RESPIRATORY CELLULAR IMPLICATIONS IN STREPTOZOTOCIN-INDUCED DIABETES: A FOCUS ON TYPE I ALVEOLOCYTES

Introduction. The escalating global prevalence of diabetes mellitus has ignited concerns, given its multifaceted repercussions that extend beyond conventional metabolic considerations. Recent scientific inquiries have firmly established a profound association between diabetes mellitus and abnormalities within the respiratory system, unveiling a nuanced interplay that transcends mere glycemic control. This study delves into the intricate and far-reaching influence of diabetes on the respiratory system, recognizing it as a vulnerable target organ susceptible to systemic repercussions. Compelling epidemiological evidence underscores a heightened prevalence of respiratory complications among individuals grappling with diabetes. This observation prompts a meticulous exploration of the pathophysiological underpinnings of this intricate relationship, spanning from molecular intricacies to clinical manifestations.&#x0D; The aim was to deepen our understanding of the impact of diabetes mellitus on the respiratory system by investigating the pathological alterations in type I alveolocytes within an experimental model of diabetes mellitus.&#x0D; Methods. In this experimental model we used 88 male Wistar rats (170-210 g). The rats were divided into three groups: Group 1 (n=10) consisted of intact rats; Group 2 (n=40) served as the control group, and Group 3 (n=38) constituted the experimental group where diabetes was induced by intraperitoneal administration of streptozotocin (Sigma, USA) diluted in 0.1 M citrate buffer with pH 4.5 at a dosage of 60 mg/kg of body weight. The control group received an equivalent volume of 0.1 M citrate buffer solution with a pH of 4.5 via intraperitoneal injection. All procedures were performed under sodium thiopental anesthesia at a dose of 60 mg/kg of body weight. Tissue samples were collected at intervals of 14, 28, 42, and 70 days post streptozotocin injection.&#x0D; For electron microscopy analysis, fragments of lung tissue were immersed in a 2.5% glutaraldehyde solution for fixation, followed by fixation in a 1% osmium tetroxide solution. After dehydration, the specimens were embedded in Epon Araldite. Sections, obtained through a "Tesla VS-490" ultramicrotome, underwent examination using a "PEM-125K" electron microscope.&#x0D; Results. Throughout the experiment, discernible changes in type I alveolocytes were observed. At 14 days, nuclei exhibited rounding or oval shape with uniform chromatin distribution. Mitochondria displayed small size, and the Golgi apparatus (GA) and granulated endoplasmic reticulum (GER) showed no significant alterations. By 28 days, nuclei adopted an oval shape, chromatin localized peripherally, and mitochondria exhibited diverse morphologies. Increased micropinocytotic vesicles indicated heightened cellular activity. At 42 days, hyperhydration became pronounced, nuclei displayed lower electron density, and cellular components showed advanced changes. By 70 days, dystrophic-destructive changes included low electron density nuclei, disorganized mitochondria, and fragmented GA and GER.&#x0D; Conclusion. This comprehensive ultrastructural analysis unveils the progressive impact of diabetes on type I alveolocytes, elucidating unique facets of pulmonary alterations over time. This study contributes to a growing body of knowledge, shedding light on the dynamic nature of pulmonary changes in diabetes mellitus, ultimately urging further exploration for a holistic understanding of its implications on respiratory health.

Influence of experimental heart failure therapy with different generations of β-adrenergic blockers on Cardiac Electrical Activity (ECG) and Autonomic Regulation of Heart Rhythm (ARHR)

Abstract: In the complex treatment of chronic heart failure (CHF) β-adrenoblockers (carvedilol, nebivolol, bisoprolol, metoprolol) are used, which due to their pharmacological properties increase the survival rate of patients, improve cardio- and haemodynamic parameters, metabolism. However, modern realities in medicine require creation of new more effective and safe β-adrenoblockers. In this regard, the potential drug Hypertirl is of interest. The aim was to evaluate the cardioprotective effect of 1-(b-phenylethyl)-4-amino-1,2,4-triazolium bromide (Hypertril) on cardiac electrical activity and autonomic regulation of heart rhythm in a model of CHF in comparison with β-adrenoblockers of different generations (Nebivolol, Carvedilol, Bisoprolol, Metoprolol). Materials and methods: Chronic heart failure was induced by 14-day administration of doxorubicin in a cumulative dose of 15 mg/kg to white mongrel rats weighing 190–220g (total 85). The investigated drugs were administered after doxorubicin course for 30 days: Hypertril at an experimentally substantiated dose of 3.5 mg/kg, Metoprolol succinate 15 mg/kg, Nebivolol 10 mg/kg, Carvedilol 50 mg/kg, Bisoprolol 10 mg/kg. At the end of drug administration under thiopental anaesthesia (40 mg/kg), electrocardiogram (ECG) and autonomic regulation of heart rhythm (ARHR) were analysed using a computer analyser CardioCom-2000plus (KAI-Medica, Ukraine). The results of the study were calculated using a standard statistical package “STATISTICA for Windows 6.0” (StatSoftInc., №AXXR712D833214FAN5), “SPSS 16.0” and “Microsoft Office Excell 2003”. Results and discussion: Hypertril administration resulted in a negative chronotropic effect, normalisation of atrial (P) and ventricular (R) spike amplitude, ST segment inversion below isoline, increased amplitude of ventricular myocardial repolarisation T. Myocardial repolarisation spike were observed, as well as normalised the duration of atrial (P) and ventricular depolarisation phase (QRS complex) to the intact value and restored the duration of electrical diastole (TR interval). Hypertril reduced systolic and diastolic myocardial dysfunction in animals with CHF. Hypertril restored autonomic mechanisms of heart rhythm regulation and balanced activity of sympathetic and parasympathetic parts of autonomic nervous system in the control of cardiac function. Conclusion: The obtained results demonstrated the undoubted advantage of the new original molecule (Hypertril) over basic β-adrenoblockers (Metoprolol, Nebivolol, Carvedilol and Bisoprolol) and experimentally justify further in-depth study to create on its basis a drug for the treatment of CHF.

Dynamics of the antioxidant-prooxidant balance of the kidney under conditions of acute blood loss of various degrees and its correction

The purpose of the study is to find out the dynamics of the antioxidant-prooxidant balance of the kidney during acute blood loss of various degrees and to evaluate the effectiveness of correction with Ringer’s lactate solution in combination with 2-ethyl-6-methyl-3-oxypyridine succinate.Material and methods. 126 male Wistar line rats weighing 0.16-0.18 kg were randomly selected for the experiment. Four experimental groups and one control group were selected. Acute blood loss of 1% of body weight was simulated in experimental group 1, acute blood loss of 2% of body weight was simulated in experimental group 2. In experimental group 3, acute blood loss in the amount of 2% of body weight was simulated, and after 1 hour, Ringer’s lactate solution was injected once into the adjacent vein in a ratio of 1:1 in the amount of blood loss volume. In experimental group 4, after simulating acute blood loss in the amount of 2% of body weight, Ringer’s lactate solution in combination with 2-ethyl-6-methyl-3-hydroxypyridine succinate at a dose of 100 mg·kg-1 was injected into an adjacent vein once. A control group of rats was only injected with thiopental sodium anesthesia. After 1, 3, and 7 days, catalase activity, the content of reagents to thiobarbituric acid were determined in the kidney, and their ratio - the antioxidant-prooxidant index (API) was calculated.Results and discussion. It was established that acute blood loss within 1-7 days of the experiment is accompanied by the accumulation of thiobarbituric acid reagents in the kidney and by a decrease in catalase activity and the value of API. Disturbances depend on the volume of blood loss and are significantly greater under conditions of acute blood loss in the amount of 2% of the mass. The injection for correction with Ringer’s lactate solution in combination with 2-ethyl-6-methyl-3-oxypyridine succinate compared to monotherapy with Ringer’s lactate solution leads to a significant decrease in the content of reagents to thiobarbituric acid in the kidney, and an increase in catalase activity and the value of API, starting from the 3-rd day of the experiment.

STAGES OF MORPHOLOGICAL CHANGES OF TARGET ORGANS IN EXPERIMENTALLY SIMULATED ACUTE PERITONITIS

One of the urgent problems in urgent abdominal surgery remains acute disseminated peritonitis (AP), the mortality rate of which remains high due to the development of multiple organ failure syndrome, despite the use of modern methods of diagnosis and treatment.The purpose of the study. To substantiate the possibility of using laparoscopic methods of treatment depending on the severity and stage of morphological changes of the target organs and the duration of peritonitis simulated in the experiment.Material and methods. The experiment used 150 sexually mature non-linear female rats, weighing 180.0-220.0 grams, which were kept on the standard ration of the vivarium of the Central Research Laboratory with maintenance of food and drinking regimes recommended by the norms of keeping laboratory animals. The animals were divided into 5 groups – a control group (30 rats) and 4 experimental groups (120 rats), which underwent HP simulation by intraperitoneal injection of 1 ml of 30 % autocal. The animals of the research group were removed from the experiment under sodium thiopental anesthesia by bloodletting on the 1st, 3rd, 5th, and 7th days. Results of the study. Morphohistochemical study of the target organs on the 1st day of the experiment: singleorgan changes were observed (examination of the peritoneum revealed its pronounced swelling). A slight vascular reaction of the basal membrane of the mucous membrane was established in the small intestine. On the 3rd day, the changes remained monooraneous with increases in infl ammatory phenomena. On the 5th day, the spread of the systemic infl ammatory response to the lungs and kidneys was confi rmed. Areas with foci of dystrophic- necrotic changes in the epithelium of the tubules were visualized in individual excretory tubules in the kidneys.In the lungs: the number of erythrocytes and fi brin increased in the cellular component. On the 7th day of the experiment, the heart and liver were involved in the process. In the heart, dystonia and paresis of microcirculatory vessels, erythrocyte stasis, and the formation of cell aggregates in the lumen of vessels were observed. In the liver, it is observed: congestion of the central veins, manifestations of venous stasis.Conclusion. Based on the stages of morphological changes of the studied organs at diff erent times, the simulation of acute peritonitis allowed us to theoretically justify a diff erentiated approach to the selection of the type of surgical interventions.

Combining laser speckle imaging and spatially modulated visible illumination to study the effect of anesthetic drugs on the optical and physiological parameters of brain

This work employs an integrated optical imaging system to evaluate the effect of anesthetic drugs on several brain parameters including absorption, scattering, oxygen saturation, and cerebral blood flow (CBF). Two anesthetic drugs, namely ketamine-xylazine (KX) and thiopental, frequently used by researchers in mouse experiments, were investigated herein. The combined system is based on spatially modulated visible light illumination and a laser speckle based-technique. With this apparatus, a series of structured light (sinusoidal patterns) with phase shifts at each of two spatial frequencies (low and high) are projected onto the surface of a mouse brain and diffusely reflected light is acquired by a camera. A six-position filter wheel, equipped with five visible bandpass filters, is placed at the output of the projector. Then, structured illumination is blocked and a laser source illuminates the tissue while the diffusely reflected light is captured by the same camera through the remaining open hole in the wheel. In this manner, visible and laser speckle images are captured and processed off-line to reconstruct the cerebral tissue features. This work's major findings are six-fold: first, over a specific range of the spectrum a difference of more than 20% was found in the absorption coefficient presumably as a sensitivity to hemoglobin. Second, a difference of more than 20% was observed between drug types in the scattering coefficient along the entire spectral range. Third, CBF varied considerably in thiopental anesthesia throughout the experiment. Fourth, oxygen saturation level remains stable in both anesthetics. Fifth, thiopental keeps blood glucose in normal range while with KX glucose level increases. Sixth, the co-registered system is capable of revealing quantitative functional contrast and may be an attractive method in a variety of biomedical applications. Overall, these findings suggest that the choice of KX or thiopental should be carefully considered as a component of study design in research utilizing animal models.

CORRECTION OF DISORDERS IN THE DIGESTIVE TRACT OF RATS WITH CHRONIC COLESTASIS WITH THE LEQUIN AND MINERAL COMPLEX

Problem. Complete or partial cessation of the flow of bile into the duodenum during cholestasis not only impairs the absorption of micro- and macroelements, fats and fat-soluble vitamins in the intestine, but also disrupts the microbiocenosis in the mucous membranes of the digestive tract. Knowledge of the mechanisms of disorders in the digestive tract caused by a decrease in the outflow of bile is important for the development of effective pathogenetic therapy. The drug Lequin (lecithin and quercetin), as well as the natural sorbent Mineralol, can be considered as such a preventive agent. Aim. To investigate the condition of the mucous membranes of the digestive tract of rats against the background of chronic cholestasis and to evaluate the effectiveness of prevention with the Mineralol and Lequin complex. Methods. The research was carried out on rats, which were divided into three groups: 1 group – intact, 2 group – rats that were modeled chronic cholestasis, 3 group – rats that were injected with Lequin (500 mg/kg) and Mineral (1 g/kg) against the background of cholestasis). Cholestasis in rats was modeled by ligation of the common bile duct under thiopental anesthesia. The drugs were administered orally to the rats of the 3rd group daily in the morning on an empty stomach for 4 months. Biochemical studies were carried out in blood serum (alkaline phosphatase, alanine aminotransferase, elastase activity), liver (elastase, acid phosphatase activity and malondialdehyde content) and mucous membranes of the oral cavity, small andlarge intestine (elastase, acid phosphatase activity and malondialdehyde content). Statistical processing of the research results was carried out using the Student-Fisher method. The main results. The conducted studies established liver parenchymal damage and cholestasis phenomena in rats with ligation of the common bile duct, which was confirmed by a decrease in alanine aminotransferase activity by 30.8% against the background of an increase in elastase activity by 42.0% and alkaline phosphatase activity by 43.6% in animal blood serum. Chronic cholestasis in rats led to the development of inflammatory processes in the liver: an increase in the activity of elastase by 24.5%, the activity of acid phosphatase by 29.8%, the level of malondialdehyde by 36.8%, along with a decrease in the antitoxic function of the liver: an increase in the activity of urease 2.2 times. Chronic cholestasis in rats caused the development of inflammation in the mucous membranes of the digestive tract: an increase in elastase activity by 28.4–41.1%, acid phosphatase activity by 19.3–45.6%, and malondialdehyde content by 55.7–127.0%. Daily prophylactic use of a complex of Lequin and Mineral drugs in conditions of chronic cholestasis in rats prevented the destruction of hepatocytes, the development of cholestatic phenomena, improved the antitoxic function of the liver, effectively prevented inflammatory processes, and the activation of lipid peroxidation in the mucous membranes of the digestive tract of animals. Conclusions. Established disorders in the mucous membranes of the digestive tract of rats with cholestasis are primarily caused by a decrease in the outflow of bile, which has antimicrobial properties, as a result of which toxic bile acids accumulate in the liver parenchyma along with a violation of the antitoxic function of the liver. The preventive effectiveness of the drugs was manifested due to the hepatoprotective, antioxidant, anti-inflammatory and choleretic effects of lecithin and quercetin, which, in combination with the sorption properties of Mineralol, made it possible to support the antitoxic function of the liver and its metabolic functions within normal limits. The conducted studies can be the basis for the use of the proposed drugs in the clinic for the treatment and prevention of cholestasis.

Effect of adenosine triphosphate on amiodarone-induced optic neuropathy in rats: biochemical and histopathological evaluation

Objective This study aims to investigate possible preventive effect of ATP on optic nerve damage caused by amiodarone in rats. Material and method Thirty albino male Wistar rats weighing between 265 and 278 g were used in the study. Before the experiment, the rats were housed at 22 °C in a 12-h light/dark cycle under appropriate condition. The rats were equally divided into five groups of six animals each: healthy group, 50 mg/kg amiodarone (AMD-50), 100 mg/kg amiodarone (AMD-100), 25 mg/kg ATP + 50 mg/kg amiodarone (ATAD-50), and 25 mg/kg ATP + 100 mg/kg amiodarone (ATAD-100). At the end of 14th day, the animals were sacrificed using cardiac puncture under deep thiopental anaesthesia, and optic nerve tissues were harvested to measure superoxide dismutase (SOD), total glutathione (tGSH), malondialdehyde (MDA), and catalase (CAT) levels. Results The MDA levels were found to be significantly higher in the AMD-50 and AMD-100 groups compared to the healthy group (p ˂ 0.001). There was also a significant difference between the AMD-50 and ATAD-50 groups, and between the AMD-100 and ATAD-100 groups regarding MDA levels (p ˂ 0.001). tGSH, SOD, and CAT levels were significantly lower in the AMD-50 and AMD-100 groups compared to the healthy group (p ˂ 0.001). ATP was found to partially inhibit amiodarone-induced optic neuropathy. Conclusion The biochemical and histopathological results of this study demonstrated that amiodarone at high doses caused more severe optic neuropathy inducing oxidative damage, but ATP could relatively antagonise these negative effects on the optic nerve. Therefore, we believe that ATP may be beneficial in preventing amiodarone-induced optic neuropathy.

Simulation of peritoneal sepsis and its treatment with serum in an experiment: peculiarities of morphological disorders of liver, spleen and kidney tissues

Sepsis develops as a normal inflammatory response to various infections. It proceeds with the picture of a complex heterogeneous syndrome, which often leads to the development of multiple organ failure. The number of affected organs correlates with mortality. Organ failure is characterized by a sequence – initially pulmonary, then renal, cardiovascular, and in the terminal stage, there is a failure of the central nervous system function. The aim of the study was to model peritoneal sepsis in an experiment, develop a therapeutic serum as an analog of anti-reticular cytotoxic serum by O. O. Bogomolets (ACS), and investigate the therapeutic properties and specific morphological changes in the liver, spleen, and kidneys of septic and treated animals. To simulate peritoneal sepsis, laboratory mice were injected intraperitoneally with a solution of 10 % of filtered fecal suspension of guinea pig (from 0.05 to 3.0 ml), the level of the toxic dose at which all mice died was determined (0.35 ml of fecal suspension) after that lethal bloodletting was administered under thiopental anesthesia. Their spleens were removed, from which therapeutic serum was made. Guinea pigs of the experimental group were injected with a suspension of the spleen of mice with an increasing dose from 0.02 to 0.2 ml after the simulation of fecal peritonitis. The optimal therapeutic dose at which the ants continued to live for 14 days was determined: 0.08 ml. Under thiopental anesthesia (50 mg/kg), their liver, kidneys, and spleen were taken for histological studies. The processing of the obtained histological specimens was carried out according to generally accepted methods. Histological preparations were studied using an SEO SСAN light microscope. It was found that under conditions of peritoneal sepsis in the liver, the lobular organization of hepatocytes was dramatically disturbed. The central veins and vessels of the portal tracts were moderately dilated and filled with blood, but the lumens of the sinusoids were practically not visualized. The sizes of hepatocytes increased sharply, contours were erased, and intercellular connections were disrupted. Histological examination of the kidney revealed a drastic decrease in the blood volume in the vessels of the arterial bed, which is visualized by the structural manifestations in the cortical layer. Collaptoid shrinkage of glomerular vessels was observed, which manifested in their sharp reduction in size. A significant part of the endotheliocytes was damaged. Examination of the spleen revealed a significant increase in the area of the red pulp due to the pronounced expansion of the sinusoids and an increase in their blood supply. The white pulp exhibited small, moderately diffuse foci of lymphocyte clusters. Follicle structures were practically not visualized. Histological examination of the liver in animals with simulated peritoneal sepsis on the background of correction with an extract from the spleen of mice revealed a moderate expansion and full blood vessels of the portal tracts and central veins. The contours of the vast majority of hepatocytes grew clear, intercellular contacts were restored. Histological examination of the kidney revealed a moderate increase in the blood volume in the vessels of the arterial bed, mainly in the cortical layer. A mild expansion and fullness of blood vessels of the glomeruli was observed, which was manifested by their increase in size. An increase in macrophage-type cells was observed in the perivascular areas. Histological examination of the spleen revealed a pretty large area of red pulp, moderate expansion and fullness of the sinusoids, and pronounced perisinusoidal edema. However, a rapid expansion of the white pulp was observed, which manifested in the formation of follicles.