Thiopental Anesthesia Research Articles

УЛЬТРАСТРУКТУРНІ ОСОБЛИВОСТІ НЕЙРО-ГЛІАЛЬНО-КАПІЛЯРНИХ ВІДНОСИН СА3 ПОЛЯ ГІПОКАМПА ЧЕРЕЗ 21 ДОБУ ЗА УМОВ ЕКСПЕРИМЕНТАЛЬНОЇ ТЕРМІЧНОЇ ТРАВМИ

Severe thermal injury leads to a number of morphofunctional changes that occur in violation of integrity or loss of skin in deep burns and endogenous intoxication on the background of which significant structural and metabolic disorders in all organs and systems of the affected organism are developed. This is manifested in a complicated symptom complex of burn disease. Changes in the central nervous system play an important role in the pathogenesis of burn injury, the nature and depth of morphofunctional changes in individual cerebral structures, in particular in the hippocampus (central organ of the limbic system, which is especially sensitive to various pathogens), after thermal lesions remain poorly studied. The aim of our study was to establish the features of submicroscopic changes in the structural components of the hippocampus after 21 days under conditions of experimental thermal injury. The experimental study was performed on white laboratory male rats. Third-degree burns were applied under thiopental-sodium anesthesia with copper plates heated in boiled water to a temperature of 97–100°C. The size of the affected area was 18–20% of the rats’ body epilated surface. Histological changes were studied on the 21st day from the beginning of the experiment. For electron microscopy, the obtained ultrathin sections were contrasted with uranyl acetate and lead citrate according to the Reynolds method. It was found that on the 21st day after the experimental thermal injury significant alternative changes in neurons, glial cells and the microcirculatory tract of the hippocampus were defined which developed on the background of excessive endogenous intoxication. Neurons are hypochromic and sharply hyperchromic, disorganized and have an altered shape; there are cell-free areas. Endoplasmic reticulum, Golgi complex, mitochondria are destructively changed. Degeneration of macroglial cells has been noticed. Their nuclei are high in electron density in which heterochromatin predominates. Destruction and lysis of organelles, lysosomes have also been found in the cytoplasm. These changes of astrocytic glia lead to impaired trophic function and the relation between astrocytes and blood capillaries. Hemocapillaries wall remodeling is manifested by the total edema, compaction of the basement membrane. The lumenal surface of the plasmolemma of endothelial cells is homogeneous, indistinctly contoured. The cytoplasm of endothelial cells is swollen, enlightened, and poor in organelles; there is a small number of micropinocytic bubbles and vesicles.

Intermittent exposure to green and white light-at-night activates hepatic glycogenolytic and gluconeogenetic activities in male Wistar rats.

Exposure to light-at-night (LAN) has been reported to impair blood glucose regulation. The liver modulates blood glucose through mechanisms influenced by several factors that include peroxisome proliferator-activated receptor gamma coactivator-1alpha (PGC-1α) and glucose-6-phosphatase (G6Pase). This study investigated the effect of intermittent exposure to green and white LAN on some hepatic glucose regulatory factors in male Wistar rats. Animals were divided into three equal groups. Group I (control) was exposed to normal housing conditions. Groups II and III were each daily exposed to either green or white LAN for 2h (7-9pm) for 14days. Body weight and blood glucose was monitored on days 0, 7, and 14. Thereafter, retro-orbital sinus blood was obtained after light thiopental anaesthesia and serum insulin was determined. Liver samples were also obtained and evaluated for glycogen, PGC-1α, and G6Pase activity. Insulin resistance was estimated using the HOMA-IR equation. Body weight and blood glucose on days 7 and 14increased in groups II and III compared to control. Hepatic PGC-1α and G6Pase increased in group II (2.33±0.31; 2.07±0.22) and III (2.31±0.20; 0.98±0.23) compared to control (1.73±0.21; 0.47±0.11). Hepatic glycogen was 71.8 and 82.4% reduced in groups II and III compared to control. Insulin in group II increased (63.6%) whiles group III values reduced (27.3%) compared to control. Insulin resistance increased in group II (0.29±0.09) compared to control (0.12±0.03) and group III (0.11±0.03), respectively. Exposure to 2h green and white LAN in the early dark phase increases hepatic glycogenolysis and gluconeogenetic activities resulting in increased blood glucose. In male Wistar rats, exposure to green but not white LAN may predispose to insulin resistance.

Changes in Morphometric Indicators of Blood Vessels of Masticatory Muscles in Simulation of Diabetic Angiopathy in Rats

Summary. Relevance. Diabetic joint damage is common in the clinic. The blood vessels of the masticatory muscles that affect the morphogenesis of diabetic arthropathy of the temporomandibular joint remain poorly studied in this pathology.
 Objective: to study the features of remodeling of blood vessels of the masticatory muscles of the temporomandibular joint in experimental hyperglycemia by quantitative morphological methods.
 Materials and Methods. The study material was 45 laboratory mature white male rats, which were divided into 3 groups. The 1st group consisted of 15 intact apparently healthy animals, the 2nd included 15 rats with one-month diabetic angiopathy, and the 3rd had 15 rats with two-month specified pathology. Diabetes mellitus was simulated by a single intraperitoneal injection of streptozotocin from Sigma at a dose of 50 mg/kg. Euthanasia of animals was performed by bloodletting under thiopental anesthesia one and two months after the start of the experiment. The blood glucose concentration was determined. Histological micropreparations were made from the temporomandibular joint and the temporal and external pterygoid muscles. Morphometric measurements of small caliber arteries (outer diameter 26-50 μm), veins and microvessels of the hemomicrocirculatory tract were performed on muscle micropreparations; relative volumes of damaged synoviocytes and chondrocytes were determined on micropreparations of joints. Quantitative indicators were processed statistically.
 Results and Conclusions. It was revealed that the gleam of arteries considerably decreases, their wall thickens, and endotheliocytes are damaged in the studied muscles with the modeled pathology. Venous vessels are dilated, full-blooded with a reduced wall thickness. The most pronounced changes in these experimental conditions are the structure of microvessels, which is characterized by narrowing of arterioles, precapillary arterioles and hemocapillaries, pronounced expansion of postcapillary venules and venules. Plethora, stasis, thrombosis, irregularity of the lumen, sacculation, and varicose expansions were noted in venous microvessels. Venous fullness led to hypoxia, disorders of trophism and metabolism, dystrophy, necrobiosis of muscle fibers and stromal structures, foci of infiltration and sclerosis. It was revealed that morphological changes in the vascular bed of the masticatory muscles affect the morphogenesis of diabetic arthropathy of the temporomandibular joint.

Peculiarities of cytolytic syndrome development with associated injury of the abdominal organs and massive blood loss in the experiment

The function of the organs responsible for detoxification processes, in particular the liver, deserves a special role in the conditions of combined trauma. It is proved that ischemia-reperfusion of the limb caused by the imposition of a tourniquet, is the cause of significant toxemia. Under such conditions, it becomes clear that the detoxification function may be impaired, especially against the background of blunt combined trauma to the abdominal organs complicated by massive bleeding. Changes in the detoxification function of the liver are indicated by markers of the cytolytic syndrome. However, there is no clear understanding of how ischemia - limb reperfusion affects the development of cytolytic syndrome.Objective of research: to study the effect of massive blood loss associated with ischemia-reperfusion of the limb on the cytolytic syndrome indicators in case of injury of the abdominal organs. Materials and methods. 100 mature albino male Wistar rats with the body weight of 190-220 grams kept on standard vivarium forage were used in the experiment. The animals were divided into the control and 3 research groups (containing 10 animals each). Arresting bleeding tourniquet was applied proximally on the lower limbs of rats from the first research group for 120 minutes under thiopental-sodium anesthesia (40 mg/kg-1), which caused development of ischemic-reperfusion. Closed abdominal injury was simulated by means of delivering two dosed blows in the region of the abdominal cavity in the second research group; massive blood loss was simulated by means of cutting the femoral vessels and bloodletting from 25 to 30 % of the circulating blood volume in the group. Injuries from the first two groups were combined in the third group. The control group of animals received anesthesia without formation of any injuries. With the aim to determine alterations of cytolytic syndrome the activity of enzymes – markers of cytolytic syndrome ALT and AST were determined by means of a unified method. The animals of the research groups were removed from the experiment under thiopental-sodium anesthesia by means of the total bloodletting from the heart.Results. Analysis of the results of the study found that cytolytic syndrome develops irrespective of the kind of injuries simulated. Thus, even during a day of post-traumatic period ALT and AST activity increases which is clearly registered in all the three research groups. Further destruction of cellular membranes increases till the 3rd day, which is manifested by a considerable increase of ALT and AST activity in the blood serum. This process continues till the 7th day. A tendency to ALT and AST decrease is observed only in the 1st group on the 7th day. Comparison of the 2nd and 3rd research groups within the time interval enables to understand that application of tourniquets on the limbs of animals with closed abdominal injury and massive bleeding deteriorates cytolytic processes.Conclusions: Simulated injuries in the research groups were associated with the occurrence of cytolytic processes possessing a tendency to develop in the period of early signs of traumatic disease. Ischemic-reperfusion syndrome of the limbs accelerates considerably the development of systemic changes with abdominal injuries and hypovolemic shock, which was manifested by a considerable increase of ALT and AST parameters in the group with abdominal trauma and massive blood loss and becoming higher than that of the control in end of the experiment.

Dynamics of connective tissue metabolites in the blood of rats of different age under hypokinesia

Topicality. Hypokinesia is a condition of insufficient motor activity of the body with limited pace and volume of movement, and is currently the fourth leading cause of endemic death in the world. It is known that hypokinesia can affect the metabolism of components of bone and cartilage, which is the basis for the development of pathological processes, but there are no biochemical markers of this process in the literature. Thus, currently, there is insufficient number of experimental scientific works devoted to the study of biochemical parameters characterizing the state of the connective tissue in the experiment to assess the impact of hypokinesia on the body, and this fact determines the relevance of the study. Aim. To study the dynamics of the main metabolites characterizing the state of the connective tissue in the blood serum in hypokinesia in rats of different ages. Materials and methods. The studies were performed on 42 white male rats aged 3 and 12 months, kept in the vivarium of the Sytenko Institute of Spine and Joint Pathology National Academy of Medical Sciences of Ukraine, Kharkiv. The conditions of hypokinesia were reproduced using a specially designed device that limited the mobility of the animal without disturbing the ventilation of the body. Animals were removed from the experiment by decapitation in Day 7 and 30 of immobilization under thiopental anesthesia. The content of sialic acids in the serum was determined by the Hess method, chondroitin sulfate – by Nemeth-Csoka method modified by L. I. Slutsky, the fractional composition of glycosaminoglycans (GAG) – by the reaction with resoquine. Results and discussion. In the study of the rat serum, it was found that in intact animals of 3 and 12 months of age the content of sialic acids was different; it was 2.94 ± 0.16 and 1.98 ± 0.09 mmol/L, respectively. After 7 days of hypokinesia, their concentration in young animals did not change, but in older animals, this figure increased. By Day 30, the serum sialic acid levels suddenly increased compared to intact animals. In animals of 12 months of age, the concentration of sialic acids in the blood serum increased by both Day 7 and Day 30 of hypokinesia, especially at the end of the experiment. Hypokinesia also showed a significant increase in serum concentrations of chondroitin sulfates on Day 7 of the experiment in animals of both age groups. At the same time, the level of these metabolites on Day 30 was at the same level. In young rats on Day 7 the fraction of GAG remained unchanged, but on Day 30 there was a significant decrease. During this period, the second fraction of GAG increased. In animals aged 12 months on Day 7 and 30, the content of most GAG fractions was lower than the values characterizing the fractional composition of GAG in intact rats. Conclusions. Thus, the determination of sialic acids, GAG fractions and chondroitin sulfates in the serum can be presented as a set of biochemical tests to assess disorders of the connective tissue metabolism while limiting the motor activity of the body. It has been found that the 30-day hypokinesia causes significant disorders of the connective tissue, and it is reflected in changes in the biochemical parameters of the serum of experimental rats.

Histological changes of the adrenal glands vessels after experimental thermal trauma and under the conditions of lyophilized xenoskin use

Burns, as a stress factor, cause circulatory and microcirculation disorders in the organs. In the scientific literature, the issues of structural reorganization of the vascular bed of the adrenal glands in thermal trauma and under conditions of correction by lyophilized xenoskin are insufficiently covered and therefore require further study. The aim of our study was to study the microscopic changes of the vessels of the adrenal glands in the dynamics after experimental thermal trauma and under conditions of correction. The experiment was performed on adult white male rats. Grade III burn (18-20 % of the skin surface) was applied under thiopental-sodium anesthesia with copper plates heated in water to a temperature of 97-1000C, on the epilated surface of the skin of the back for 10 seconds. One day after the thermal injury, an early necrectomy was performed and the wound was covered with a substrate of lyophilized xenoskin. Animal decapitation and microscopic changes were studied at days 7, 14, and 21 of injury modeling. The manufacture of histopreparations was carried out according to conventional methods. Histological examinations after experimental thermal trauma and correction of xenoskin substrate on the 7th day showed that already in this period of the experiment there were less pronounced destructive changes in the structure of the vessels of the adrenal glands. Some blood vessels were unevenly filled and dilated. The hemocapillaries of the microcirculatory tract are blood-filled, the vessel wall is destructively altered, and perivascular edema is present. On the 14th day of the experiment, moderately blood vessels were observed. The wall of large vessels is slightly swollen, partially thickened, hemocapillaries are moderately blood-filled, with dilated lumens and continuous endothelial lining. Under the conditions of application of the substrate of lyophilized xenoskin on the 21st day of the experiment, the structural organization of the vascular bed of the adrenal glands was significantly better compared to the previous terms of the study. Most vessels had evenly dilated lumens and moderate blood supply, and wall edema was insignificant. The walls of microvessels are preserved, well structured, endotheliocytes without severe damage, without signs of infiltration and edema. Thus, the results of microscopic examination showed that early necrectomy, and subsequent closure of the wound with a substrate of lyophilized xenoskin, creates optimal conditions for regenerative processes in both burns and vascular bed of the adrenal glands. On the 7th day after the burn and under conditions of correction, the study of the vascular bed of the adrenal glands showed a moderate degree of destructive changes in the vascular bed of the adrenal glands and the initial signs of their regeneration. In the late terms (14-21 days) of the experiment the relative normalization of cellular parenchymal components of the organ was established against the background of restoration of the structural organization of the microcirculatory tract.

Microscopic changes of the pancreas vessels in the dynamics after experimental thermal injury

The problem of burns is relevant in the world and in Ukraine. Thermal injury is not limited to local tissue changes, but causes significant morphofunctional disorders in the organs and systems of the affected organism, the result of this factor is a violation of blood supply to organs, including the pancreas. The aim of this work was to establish the histological condition of the vessels of the pancreas after experimental thermal injury. The experiments were performed on 40 adult white rats. Burns were applied under thiopental-sodium anesthesia with copper plates at a temperature of 96-97℃ on the epilated surface of the skin of the back of animals for 15 seconds (the lesion area was 18-20 % of the body surface). Decapitation of animals was performed on 1, 7, 14 and 21 days of the experiment. Pieces of the pancreas were taken for histological examination and processed according to generally accepted methods. Histological sections 5-6 μm thick were stained with hematoxylin-eosin, impregnated with silver by the Gordon-Sweets method, as well as staining by the tricolor method of MSB (ORB). Histological specimens were examined and documented using an SEO SCAN light microscope and a Vision CCD Camera. In the early stages of the experiment revealed a violation of the blood supply to the body. The lumens of most arteries were found to be dilated, and the wall thickness increased due to hypertrophy of the smooth muscle cells of the media. Leukocyte infiltration was determined in the perivascular spaces. The lumens of the small and middle veins were dilated and filled with blood, with parietal thrombi in some of them. Capillaries are characterized by a moderate expansion of their lumens, stasis. There was swelling of the paravasal spaces. Significant destructive-degenerative, inflammatory and sclerotic changes were observed in the later stages of the experiment. The walls of the vessels were moderately swollen. Smooth myocytes of the arterial media were hypertrophied, the cytoplasm was vacuolated, and contained pyknotically altered nuclei. Adventitia of vessels, especially veins, was swollen and fibrous. Occasionally there were violations of the wall of the components of the microcirculatory tract, which was accompanied by hemorrhage. There are single thrombosed vessels, spasmed arterioles, hemocapillaries with dilated lumens. The greatest changes, especially on the 21st day, were the adventitial membrane with the growth of collagen and reticular fibers. The blood supply to the vessels was moderate. Vessels of small diameter and hemomicrocirculatory tract were also significantly altered, their wall is deformed, thin, homogeneous. The lumens are dilated, with the formation of blood clots. Thus, in the early stages of the experiment there are adaptive-compensatory processes and initial destructive changes in the walls of the vessels of the organ, manifested by stasis, thrombosis, edema, deformation, infiltration of the wall and perivascular space. In the late period, significant destructive-degenerative changes in the structure of the walls of the vessels of the pancreas, accompanied by hemorrhage, inflammation, sclerosis of the wall and edema of the adventitia. These changes lead to a deterioration of blood supply and, consequently, a decrease in the functional activity of the body.

Reorganization of cerebellar cortex structural components one day after experimental thermal injury

Thermal injury is a lesion caused by thermal agents. With deep and large burns, the course of this type of injury becomes a general disease of the whole organism with the involvement of vital organs. The skin is the first to perceive the action of this exogenous factor, and secondarily there are changes in all body systems, in particular in the cerebellum. The aim of our study was to establish micro- and submicroscopic changes in the structural components of the cerebellar cortex one day after the experimental thermal injury. Simulation of experimental thermal injury was performed on white laboratory male rats. Grade III burns were applied under thiopental-sodium anesthesia with copper plates heated in boiled water to a temperature of 97-100°C. The size of the affected area was 18-20 % of the epilated body surface of rats. The cerebellum was collected after one day, further processing of the material for micro- and submicroscopic examination was performed according to accepted methods. Histological specimens were stained with methylene blue, and for electron microscopy the obtained ultrathin sections were contrasted with uranyl acetate and lead citrate according to the Reynolds method. At the micro- and submicroscopic levels, it was found that 1 day after the experimental thermal injury, reactive changes are observed in neurons, microcirculatory tract and glial cells. Thus, one day after the thermal trauma of the skin, the experimental animals showed the initial violations of the blood-brain barrier of the cerebellar cortex, which are adaptive-compensatory in nature. Initial, destructive, changes in neuro-glial-capillary relations, which occur primarily on their damage to the walls of hemocapillaries and are manifested by a violation of the micro- and ultrastructure of the main cells of the cerebellum – Purkinje, neurocytes of molecular and granular layers, neuroglyocytes, with peri-gliocyocytes were found.

Alterations of bile secretion indicators with blunt multiple abdominal injury and underlying ischemia-reperfusion of the limbs

It is known that ischemia-reperfusion has a significant negative impact on combined trauma. It is clear that there is an impression and dysfunction of internal organs against the background of activation of systemic pathological processes, especially those responsible for detoxification processes. However, these processes are little studied in detail, in particular regarding the violation of bile secretion. There is no clear data, and evidence of exactly how ischemia-reperfusion affects liver function in the case of blunt combined trauma to the abdominal organs and how much bile production deteriorates. This is what has become the subject of research.Objective of research: to study dynamic peculiarities of the liver bile secreting function in response to abdominal injury complicated by hypovolemic shock and ischemic-reperfusion syndrome. Materials and methods. 80 mature albino male Wistar rats with the body weight of 190-220 grams kept on standard vivarium forage were used in the experiment. The animals were divided into the control and 3 research groups (containing 8 animals each). Arresting bleeding tourniquet was applied proximally on the lower limbs of rats from the first research group for 120 minutes under thiopental-sodium anesthesia (40 mg/kg-1), which caused development of ischemic-reperfusion syndrome. Closed abdominal injury was simulated by means of delivering two dosed blows in the region of the abdominal cavity in the second research group; hypovolemic shock was simulated by means of cutting the femoral vessels and bloodletting from 20 to 22 % of the circulating blood volume in the group. Injuries from the first two groups were combined in the third group. The control group of animals received anesthesia without formation of any injuries. Bile secreting function of the liver was studied on the basis of identification of the content of total, conjugated/direct and unconjugated/indirect bilirubin in bile, and the volume of bilirubin conjugation degree in bile. The animals of the research groups were removed from the experiment under thiopental-sodium anesthesia by means of the total bloodletting from the heart.Results. A considerable disorder of the bile secreting function of the liver was found in response to simulated injuries: bile secretion rate in the first research group in comparison with the control one achieves minimal values in 3 days and remains on the same level till the seventh day of the post-traumatic period. In the second group the parameter does not change during 1-3 days, though it decreases sharply in seven days. Unidirectional decrease was found in the third group till the seventh day of the experiment.The data obtained are clearly indicative of a negative effect of ischemic-reperfusion syndrome on the bile-forming and bile secreting functions of the liver. Effect of the liver function was found both with isolated action and with multiple injuries of the abdominal organs, when bilateral compromised syndrome is observed. Conclusions: The study proved that the combined blunt trauma of the abdominal organs negatively affects the process of bile formation. Also, in the presence of ischemia-reperfusion of the limb, pathological processes are characterized by a tendency to increase significantly.

The effect of cranioskeletal trauma complicated by blood loss on the functional state of the liver in the early period of traumatic disease in rats with different resistance to hypoxia and their correction

Introduction. Traumatic events are considered to be one of the current problems in modern urban society. Beyond being the immediate cause of the death of the injured, the development of multiple organ failure syndrome is a perilous complication of severe multiple and combined trauma. An experimental cranioskeletal trauma is known to be accompanied by internal organ dysfunction. Nonetheless, the development of organ dysfunction in terms of hypoxia resistance in the presence of cranioskeletal trauma remains insufficiently studied.The objective of research: to establish the dynamics of the functional state of the liver in the presence of cranioskeletal trauma, complicated by blood loss in rats with different hypoxia resistance in the early period of traumatic disease and evaluate the efficacy of Thiocetam in the correction of identified abnormalities.Material and methods: The experimental studies were conducted on 196 white non-linear male rats weighing 180-200g. Initially, an individual resistance to hypoxia of the rats was estimated, and the animals demonstrated high- and low- resistance value to hypoxia (HR and LR, respectively) were selected for the further study. The HR- and LR-rats were separately divided into 4 groups: control and three experimental once. In the first experimental groups the HR-and LR-animals were induced the cranioskeletal trauma under thiopental sodium anesthesia (40 mg·kg-1); the animals of the second experimental groups were inflicted the acute blood loss of 20-22% of circulating blood volume; in the third experimental groups, the HR and LR animals were subjected to an acute blood loss following the cranioskeletal trauma and administered the intraperitoneal injection of the Thiocetam at a dose of 250 mg·кg-1 of body weight once a day for correction. The bile excretion function of the liver was analyzed in the controls and experimental groups of animals 1, 3 and 7 days after trauma.The results and discussion. The conducted studies indicated the higher rate of bile excretion in the control group of LR-rats compared to the control of HR-rats. The rate of bile excretion was occurred decreased in both HR- and LR-rats under the influence of the cranioskeletal trauma. However, the degree of decrease in the studied parameter of the LR-rats was considerably greater than the corresponding value of the HR-rats. An additional blood loss model resulted in more marked abnormalities, particularly in the experimental group of LR-rats. The 7-day administration of Thiocetam led to the significant decrease in the abnormal bile excretion rate in the experimental groups of HR- and LR-rats compared to the animals without corrective medication. The analysis of deviation degree of the studied parameter in the animals administered Thiocetam in relation to the animals without correction evidenced better efficacy of the medication in the experimental group consisted of LR-rats compared to the group of HR-rats, especially under the conditions of additional acute blood loss model. Consequently, Thiocetam is able to compensate less-developed defense mechanisms of LR-rats compared to the HR-rats, which should be taken into consideration in real-life clinical practice settings in the comprehensive treatment of the injured with cranioskeletal trauma complicated by blood loss considering the possibility to determine their resistance to hypoxia.Conclusions. The rate of bile excretion as a basic indicator of the functional state of the liver in the intact LR-rats is found to be substantially higher than in HR-rats. The degree of decrease in the rate of bile excretion is significantly greater in LR-rats after 7 days of the post-traumatic period under the influence of cranioskeletal trauma complicated by acute blood loss. The administration of Thiocetam is accompanied by a marked positive effect on the rate of bile excretion in the HR- and LR-rats, but the degree of increase in the studied parameter is considerably greater in the experimental group of LR-rats.

Оригінальний спосіб вилучення та фіксації гіпофіза статевозрілих щурів для приготування якісних постійних гістологічних препаратів

The issue of high-quality creating histological preparations is an urgent issue of histology and pathological anatomy, experimental medicine and biology. The pituitary gland belongs to the smallest in mass and size of the endocrine glands in both humans and animals, it is an endocrine gland and occupies one of the central places in the endocrine regulation of the body's vital activity. The study of the features of the removal of the pituitary gland from Turkish saddle of the sphenoid bone of the skull of rats remains an important aspect of modern morphology. The purpose of the work was to develop an original technique for improving the first (removal) and second (material fixation) stages of creating histological preparations of the rat pituitary gland for morphological, morphometric, and immunohistochemical medico-biological experimental studies. Material and methods. The development of the methodology was carried out on 200 white sexually mature rats of different sex weighing 250-300 g at the age of 7-8 months in accordance with the National and European bioethical standards. Results and discussion. Modification of the technique of extracting the pituitary gland of rats includes the introduction of the animal into thiopental anesthesia, decapitation, separation of the skin and muscular integument of the head, resection of the occipital bone of the skull, exposure and removal of the brain, identification of the pituitary gland, followed by removal of the pituitary gland with a single complex (block) together with the pituitary fossa Turkish saddle and fragments of the sphenoid bone adjacent to it. The gland was fixed with a reduced concentration of a 5% solution of buffered neutral formalin. After 15-18 hours from the beginning of fixation, the complex was temporarily removed from the fixator, and the pituitary gland was removed from the pituitary fossa of Turkish saddle using eye scalpel and eye forceps. The organ had a well-fixed and compacted capsule, which prevented unnecessary trauma and fragmentation of the organ during removal. The removed pituitary gland was again immersed in a 5% solution of neutral buffered formalin for 2-3 hours. After the end of the fixation period, the pituitary gland had a well-fixed structure and was subject to the following standard stages of making permanent histological specimens

Ліпідний склад плазми крові щурів за дії етилтіосульфанілату і хром (VI)-індукованої токсичності

Ethylthiosulfanylate (ETS) is synthetic organosulfur compound and belongs to the class of thiosulfonates, which are the structural analogues of natural BACs of plant origin. Thiosulfonates and ETS in particular affect the regulation of pro/antioxidant status, lipid and protein metabolism in the tissues of laboratory rats. Cr(VI) compounds are characterized by potent prooxidant properties and high toxicity for cells of living organisms. The toxic effect of Cr(VI)-induced oxidative stress is accompanied by lipid metabolism disorders and the correction methods have not been sufficiently studied. The aim of our study was to investigate the effect of ETS on some indicators of lipid metabolism in blood plasma of rats under the action of K2Cr2O7-induced toxicity. Animals were divided into 7 groups. Animals of group I injected daily intraperitoneally with 150 μl of physiological solution for 7 days. Experimental groups III and IV were administered daily intraperitoneally with K2Cr2O7 in a dose of 2.5 mg Cr(VI)/kg body weight, for 7 (group III) and 14 days (group IV). Rats of group II received intragastric injection of 1000 μl of oil daily for 14 days, than animals were injected daily intraperitoneally with 150 μl of physiological solution for 7 days. Experimental group V was intragastrically injected with ETS oily solution at a rate of 100 mg/kg of body weight daily for 14 days, than animals were injected intraperitoneally 150 μl of physiological solution daily for 7 days. Animals of groups VI and VII were intragastrically administered with ETS oily solution at a rate of 100 mg/kg of body weight daily for 14 days, than animals were injected intraperitoneally daily K2Cr2O7 in a dose of 2.5 mg Cr(VI)/kg body weight, for 7 (group VI) and 14 days (group VII). Rats were decapitated under thiopental anesthesia, after which blood was taken and divided into erythrocytes and plasma. The Cr(VI) action for 7 (group III) and 14 days (group IV) led to an increase in the content of total lipids and triglycerides in the blood plasma of rats, but the percentage of nonesterified cholesterol decreased. ETS partially compensates the Cr(VI)-induced toxicity by reducing the intensity of total lipids (groups VI, VII) and triglycerides (group VI) accumulation.

Regional anesthesiological support for the implementation of surgical interventions in the abdomen of dogs

Widespread use of surgical interventions in the abdomen of dogs determines the relevance of the study. The research tested the influence of therapeutic epidural blockade with 0.2% bupivacaine after surgical interventions on the background of potentiated local anaesthesia on the body of clinically healthy dogs undergoing ovariohysterectomy. In the control group postoperative analgesia was performed with the nonsteroidal anti-inflammatory drug rimadyl on the background of potentiated thiopental anaesthesia. Wound healing rates were determined in 10 animals of both control and experimental groups; blood counts and the level of inflammatory response were indicated in 5 animals of each group. The dynamics of reparative processes and complete wound healing in dogs of the experimental group occurred earlier than in animals of the control group, which confirmed the fact that the use of bupivacaine had a beneficial effect on tissue regeneration. The results of morphological and biochemical parameters of blood in animals of the experimental group manifested only an increase of leukocytes by 1.58 times, fibrinogen by 2.02 times, while in dogs of the control group the level of erythrocytes, haemoglobin and hematocrit decreased, leukocytes increased by 1.74 times, platelets decreased by 2.87 times and fibrinogen increased by 1.72 times. Among the stress response markers in the control group there was a 1.94 times increase in glucose level after surgery. In the experimental group there was a decrease in the intensity of the inflammatory reaction in dogs, accompanied by an increase of anti-inflammatory interleukin 4 (IL-4) only on the seventh day – 1.86 times, while in the control group the level of anti-inflammatory interleukin 1 receptor antagonist (IL-1RA) after three days increased by 2.30 times, after seven days – by 2.48 times, and the level of proinflammatory interleukin 6 (IL-6) after surgery increased by 2.57 times. It will be promising to conduct further research on the influence of therapeutic blockades with bupivacaine in animals in cases of various surgical interventions and pathological conditions, which will lead to faster recovery of animals and alleviate pathological processes.

Study of the intensity of microbial contamination and nonspecific immunity under the experimental diabetes after using different ways of fixing the wound edges

Diabetes mellitus and its complications are one of the most important causes of death. As a consequence of hyperglycemia in patients with diabetes there is an increased risk of concomitant diseases, and one of the important consequences of diabetes is impaired ability to repair. Healing disorders in diabetes are the result of complex pathophysiology involving vascular, neuropathic, immune and biochemical componentsAim of research: was to analyze changes in inflammation markers, microbial contamination and nonspecific immunity in skin homogenate of rats with diabetes after using sutures and skin glue.Material and research methods. Experimental studies were performed on 130 white outbred adult male rats, weight from 240 to 320 g. Diabetes mellitus was induced by using streptozotocin (Sigma, USA) (intraperitoneally - 65 mg / kg) with previous (15 minutes) injection of nicotinamide (intraperitoneally - 230 mg / kg). On the background of obesity, which was caused by 4 weeks of keeping animals on a high-fat diet. The development of diabetes mellitus 2 was confirmed by determining the concentration of glucose in the blood using a glucometer BAYER Contour Next (Germany). Animals of all groups (I-IV) under thiopental anesthesia (40 mg / kg body weight of rats) were made full-layer rectilinear incisions, 2 cm long, in the anterior-lateral region of the abdomen. All animals were divided into 4 experimental groups: І group (30 rats) – healthy rats, wound edges were sutured with Vicryl 5/0 thread (ETHICON, Inc. and Johnson & Johnson company (USA); ІI group (30 rats) – healthy rats, fixing of the wound edges was performed by applying skin glue Dermabond (ETHICON, Inc. and Johnson & Johnson company (USA)); ІII group (30 rats) – rats with diabetes mellitus, wound edges were sutured with Vicryl 5/0 thread; IV group (30 rats) – Rats with diabetes mellitus, fixing of the wound edges was performed by applying skin glue Dermabond. For control, all the results were compared with those of intact animals (10 rats). Animals were removed from the experiment on 3, 7, 28 days after surgery under thiopental anesthesia (90 mg / kg body weight of rats).The degree of contamination of the wound with opportunistic and pathogenic microflora determined by the level of urease activity using Nessler's reagent. Lysozyme activity was determined by bacteriolytic method, using as a substrate a suspension of bacteria Micrococcus lysodeicticus.Results. The obtained results prove that the wound process under conditions of streptozotocin diabetes mellitus in rats leads to a significantly greater contamination of the wound with pathogenic and opportunistic microflora and a decrease in nonspecific immunity, compared with healthy animals. Also, different effects of suture materials on the indicators of microbial contamination and immune protection were found: significantly larger deviations from the norm were found in groups of rats, where we used surgical sutures to fix the wound edges. When comparing the intensity of microbial contamination and nonspecific immunity in the long term (28 days) in the homogenate of the skin of rats with diabetes mellitus and the use of skin glue, the level of urease was 27.3% lower and the level of lysozyme 7.0% lower than in III group where we used surgical sutures.Changes in the activity of the antimicrobial enzyme lysozyme allow to assess the state of nonspecific immunity in the studied tissues. Many authors note a decrease in lysozyme activity in various bioliquids and tissues in diabetes mellitus. The results of our studies prove a significant decrease in the activity of lysozyme in the homogenate of postoperative skin wounds of rats with diabetes.Conclusion. Wound process in rats with experimental diabetes mellitus leads to reliable contamination of the wound with pathogenic and opportunistic microflora, as evidenced by increased urease activity, and reduced nonspecific immunity, characterized by reduced lysozyme activity in the homogenate of postoperative wound/scar tissue in relation to intact animals during all the terms of observation.In the remote period (28 days) in the skin homogenate of rats with diabetes where skin glue was used for wound closure, the level of urease was 27.3% lower and the level of lysozyme was 7.0% lower than that of rats with diabetes, where surgical sutures were used.

Violations of antioxidant-prooxidant spleen index and ceruloplasmin content in serum during acute blood loss, complicated by ischemia-reperfusion of limb and the effectiveness of its correction with carbacetam

Antioxidant-prooxidant imbalance is one of the key mechanisms of acute blood loss and ischemic-reperfusion syndrome. Under these conditions, the level of ceruloplasmin (CP) plays an important role, enhancing antioxidant protection and promoting hematopoiesis, exhibiting ferrooxidase action. However, in this case, the state of antioxidant-prooxidant balance in the spleen and the content of CP in the serum is studied insufficiently, thus there is no data on the effectiveness of carbacetam, which revealed a pronounced protective effect on antioxidant protection of the spleen.Objective of research: to determine the dynamics of the antioxidant-prooxidant index of the spleen and the content of CP in the serum in acute blood loss complicated by ischemia-reperfusion of the limb, and to evaluate the effectiveness of carbacetam in correction of the identified disorders.Materials of the research and their discussion. 108 nonlinear male rats weighing 200–220 g were used in the experiments. All experiments were performed under thiopental-sodium anesthesia. In animals, limb ischemia-reperfusion and acute blood loss were modeled separately and in one of the groups these injuries were combined. In a separate group, the detected disorders were corrected with carbacetam. After 1 and 2 hours, as well as after 1, 7 and 14 days in the spleen of experimental animals was determined content of reagents to thiobarbituric acid and catalase activity, and according to these results the antioxidant-prooxidant index (API) was calculated and was determined the content of CP in serum.Results of the research and their discussion. Modeling of acute blood loss, complicated by ischemia-reperfusion of the limb, contributed to a decrease in the value of API in the spleen, which indicates a shift in antioxidant-prooxidation balance towards the dominance of prooxidant mechanisms. Under these conditions, the content of CP in the blood serum increases significantly, which on the one hand compensates the lack of antioxidant protection, on the other hand indicates on the activation of the mechanisms of the body's systemic response to inflammation. The use of carbacetam helped to shift the antioxidant-prooxidant balance of the spleen towards the dominance of antioxidant mechanisms and to reduction of the CP content in the serum, which indicates on a pronounced antioxidant and anti-inflammatory effect of the drug, as well as its prospects in cases connected with hemic hypoxia and ischemia-reperfusion syndrome.Conclusions: Complication of acute blood loss by limb ischemia-reperfusion contributes to the shift of antioxidant-prooxidant balance of the spleen towards the dominance of prooxidant mechanisms with a simultaneous increase in the content of CP of serum. The use of carbacetam helps to increase the value of API and reduce the content of CP in the serum.

Dynamics of violations the absorption function of the small intestine in conditions of acute blood loss complicated by ischemia-reperfusion of the limbs and their correction

Introduction. In the structure of modern injuries in wartime and peacetime the share of gunshot wounds increased significantly. The timeliness of applying a tourniquet is one of the main methods of saving lives on the battlefield. The patterns of formation of enteral insufficiency syndrome in conditions of acute blood loss complicated by ischemia-reperfusion of the limb have not been studied enough. There are no data on the features of violations of the absorption function of the small intestine in these conditions and the effectiveness of correction with Carbacetam, which is able to reduce the manifestations of acute hemic hypoxia and ischemic-reperfusion syndrome.The objective of research: To establish the peculiarities of the absorption function of the small intestine in the pathogenesis of acute blood loss, ischemia-reperfusion of the limb and their combination. To evaluate the effectiveness of Carbacetam in the correction of identified abnormalities.Materials and methods. The experimental studies were conducted on 108 white nonlinear male rats weighing 200-220 g, which were housed in standard vivarium conditions. All animals were divided into five groups: control and four experimental. Under thiopental-sodium anesthesia in the first experimental group was simulated ischemia-reperfusion of the limb, in the second group – acute blood loss, and in the third group – these injuries were combined. In the fourth experimental group, animals with acute blood loss and ischemia-reperfusion of the limb were intraperitoneally administered Carbacetam at a dose of 5 mg per kilogram of animal weight. After 1 and 2 hours, as well as after 1, 7 and 14 days studied the absorption function of the intestines by D-xylose test.The results and discussion. Simulation of limb ischemia-reperfusion compared with the control group causes a decrease in the content of D-xylose in the urine after 1 day of the reperfusion period. Also, acute blood loss is accompanied by a significant decrease in this indicator after 1 day. Until the end of the experiment the rate increases, but up to 14 days does not reach the level of control. The combination of acute blood loss and ischemia-reperfusion of the limb causes a decrease in the content of D-xylose in the urine starting from 1 hour of the experiment, which reaches a minimum after 1 day and remains at the same level until 14 days. In these terms the rate was the lowest compared to other studied groups. The use of Carbacetam for corrective purposes in rats with acute blood loss complicated by ischemia-reperfusion of the limb, compared with animals without correction significantly reduces the detected abnormalities after 7 days and more after 14 days of use.Conclusions. Complications of acute blood loss by ischemia-reperfusion of the limb causes a significant decrease in the absorption function of the small intestine after 1 day of the experiment. The intensity of disorders was significantly higher compared to other experimental groups. The use of Сarbacetam for 7-14 days in the reperfusion period in animals with acute blood loss complicated by limb ischemia-reperfusion, compared with animals without correction, causes a significant increase in the absorption function of the small intestine. This indicates the protective effect of the drug under the conditions of simulated pathology.

The effect of Hippophae rhamnoides L. extract on acrylamideinduced brain injury in rats.

ABSTRACTPurpose:Reactive oxygen species (ROS), interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) have been shown in the pathogenesis of acrylamide neurotoxicity. Hippophae rhamnoides L. extract (HRE) has a cytoprotective effect by stabilizing the production of ROS, IL-1β and TNF-α. The objective of the article was to investigate the effect of HRE on acrylamide-induced brain damage in rats biochemically and histopathologically.Methods:To the HRE+acrylamide only (ACR) group (n=6) of the animals, HRE was administered orally at a dose of 50 mg / kg into the stomach by gavage. The same volume of solvent (olive oil) was administered orally to the ACR (n=6) and healthy (HG) (n=6) groups. One hour after HRE administration, acrylamide was given orally at a dose of 20 mg/kg to HRE+ACR and ACR groups in the same way. This procedure was repeated once a day for 30 days. At the end of this period, brain tissues extracted from animals killed with 50 mg/kg thiopental anesthesia were examined biochemically and histopathologically.Results:It has been shown that HRE prevents the increase of malondialdehyde (MDA), myeloperoxidase (MPO), IL-1β and TNF-α with acrylamide and the decrease of total glutathione (tGSH) and glutathione reductase (GSHRd) levels in brain tissue.Conclusions:HRE may be useful in the treatment of acrylamide-induced neurotoxicity.

Oxygen-dependent oxidation of proteins in the pathogenesis of spleen injury caused by acute blood loss, complicated by ischema-reperfusion of the limb and the efficiency of their correction by carbacetam

Activation of free radical processes is one of the key mechanisms of acute blood loss and ischemic-reperfusion syndrome. The spleen plays an important role in adaptive-compensatory processes during the blood loss. However, the state of oxidative modifications of proteins (OMP) in the spleen has not been studied enough. There are no data on the effectiveness of carbacetam in these conditions, that showed a pronounced protective effect on the enzyme link of the spleen antioxidant protection.Objective of research: to study the role of oxygen-dependent protein oxidation in the pathogenesis of spleen involvement during and after acute blood loss complicated by limb ischemia-reperfusion, and the effectiveness of its correction with carbacetam.Materials and methods. The experiments were performed on 108 nonlinear male rats weighing 200–220 g. All experiments were made using thiopental-sodium anesthesia. In separate groups of animals were simulated limb ischemia-reperfusion and acute blood loss as well as these combined injuries. In a separate group, the detected disorders were corrected with carbacetam. After 1 and 2 hours, as well as after 1, 7 and 14 days in the spleen of the experimental animals was determined the content of oxidative modifications of protein (OMP). The spleen was taken for additional investigation and the content of neutral (OMP370) and basic (OMP430) was determined in its homogenate.Results. Modeling of acute blood loss, complicated by ischemia-reperfusion of the limb, contributed to the increasing of the processes of uncontrolled oxidative modification of proteins in the spleen. The content of OMP370 rised significantly in the groups of animals, in which only ischemia-reperfusion of the limb or acute blood loss were simulated beginning from 2 h of the experiment. The content of OMP430 exceeded the base level in these experimental groups after 1 and 14 days. Violations of the studied indicators reached a maximum after 1 day, decreased to day 14, but did not reach the level of control group. Carbacetam had a protective effect on the intensity of OMP370-430 after 7 and 14 days of its usage, which puts carbacetam in a line of promising agents of complex therapy of spleen disturbances caused by acute blood loss and ischemic-reperfusion syndrome.Conclusions: Complication of acute blood loss by ischemia-reperfusion of the limb causes intensification of OMP in the spleen, that, compared with the control is an increase in the content of aldehyde and ketone derivatives of neutral and basic nature throughout the whole experiment with its maximal level at day 1. The use of carbacetam significantly reduces the intensity of OMP in the spleen compared to the group of animals that had no additional drug correction.

Study of morphological changes in the kidney during modeling of ischemic-reperfusion injuries of the limb and massive blood loss

The article presents the features of morphological disorders of liver tissue on the background of limb ischemia-reperfusion and massive blood loss. The aim of the work was to establish the presence of structural changes and the severity of morphological disorders of internal organs, remote from the primary place of ischemia-reperfusion during modeling the pathological process. The experiment was performed on 33 adult nonlinear white rats weighing 200-250 g, which were on the standard vivarium diet. Experimental animals were simulated with ischemic-reperfusion syndrome of the lower extremity and massive blood loss. Collection of materials was performed 1 h after intervention or release from the tourniquet and on the 1st, 3rd, 7th and 14th days. All interventions were performed under thiopental-sodium anesthesia (40 mg/kg body weight). Mostly structural violations were expressed as disturbance of blood supply of little and middle caliber blood vessels and also in initial dystrophic changes. During comparison of groups, depending on the severity, it was shown that both the isolated use of a tourniquet and massive blood loss had long-term, systemic consequences, however, more expressed in EG-2; single vascular glomeruli were shrunked, endothelial cells of arterioles were slightly damaged. On the 7th and 14th days changes in the structure of EG-1 were mostly absent, although in EG-2 the epitheliocytes of the outer layer of the capsule remained flattened, retained full blood vessels of the venous bed in the interstitium, and the vast majority of epitheliocytes of the excretory tubules were at different stages of hydropic dystrophy with partly desquamation of the epithelium in the gaps of the tubules. Also, the basal membranes of the tubules were not completely visualized, which indicates deep damage in the structures caused by acute ischemia as a result of bleeding. Thus, as combat trauma (blood loss) is in itself a life-threatening factor, the use of a tourniquet due to the development of ischemic-reperfusion process can complicate the course of the primary affection. Knowledge of the periodization of traumatic disease on the background of this pathology is important for the development of sanogenic effects in order to minimize this pathogenic factor.

Dynamics of endogenous intoxication indicators in condition of acute blood loss complicated by ischemia-reperfusion of the limb, and their correction by carbacetam

Introduction. In the structure of injuries from gunshot wounds dominate the limb injuries that are often complicated by massive bleeding from the main vessels. The only means of rescue on the battlefield is a timely application of a tourniquet. This ensures complete exsanguination of the limb. The duration of exsanguination should not exceed 2 hours. It is proved that in conditions of acute blood loss complicated by ischemia-reperfusion of the limb, there are prerequisites for the development of dysfunction of internal organs, which in turn exacerbates dysmetabolic and functional disorders, stimulates endotoxicosis and may be complicated by multiple organ failure. However, the patterns of formation of the syndrome of endogenous intoxication in the conditions of ischemia-reperfusion of the limb, acute blood loss and their combinations have not been studied. There are no data on the effect of Carbacetam on these processes. It is known that Carbacetam has proven itself to reduce the manifestations of dysfunction of internal organs in the conditions of ischemic-reperfusion syndrome of the limb.The objective of research: To establish the dynamics of endogenous intoxication in the conditions of ischemia-reperfusion of the limb, acute blood loss and their combination. To evaluate the effectiveness of Carbacetam in the correction of identified abnormalities.Materials and methods. The experimental studies were conducted on 108 white nonlinear male rats weighing 200-220 g, which were housed in standard vivarium conditions. All animals were divided into five groups: control and four experimental. Under thiopental-sodium anesthesia in the first experimental group was simulated ischemia-reperfusion of the limb, in the second – acute blood loss, and in the third – these injuries were combined. In the fourth experimental group, animals with acute blood loss and ischemia-reperfusion of the limb were intraperitoneally administered Carbacetam at a dose of 5 mg per kilogram of animal weight. After 1 and 2 hours, as well as after 1, 7 and 14 days in the blood serum was determined the content of middle-mass molecules (MMM254, MMM280).The results and discussion. It was found that ischemia-reperfusion of the limb in comparison with the control group is accompanied by a significant increase of the MMM254 fraction in the blood serum content. From the first hour of the experiment with a maximum after 1 day and normalization of the index up to 14 days. The content in the blood serum of the MMM280 fraction became significantly higher, from the control only after 1 day with subsequent normalization in the following terms of the experiment. After simulation of acute blood loss, both studied fractions of MMM increased from 1 hour of the experiment, reached a maximum after 1 day and did not return to the control level after 14 days. For the first time it was found that two-hour ischemia-reperfusion of the limb significantly aggravates the course of acute blood loss, which revealed a significantly higher content in the serum of fractions MMM254 and MMM280 compared to other experimental groups in almost all periods of the experiment. Due to the use of Carbacetam, the content of both studied fractions of MMM in the serum compared with animals without correction was significantly reduced after 7 and 14 days of the experiment. Although the studied parameters did not return to the level of the Carbacetam control group, it can be considered a pathogenetically justified means of reducing the level of endotoxicosis in the conditions of ischemia-reperfusion of the limb and acute blood loss.Conclusions. Simulation of limb ischemia-reperfusion is accompanied by accumulation of MMM254-280 fractions with a maximum after 1 day of the experiment and normalization up to 14 days. Acute blood loss causes an increase in the serum content of fractions MMM254-280 from 1 hour of the experiment, which up to 14 days do not reach the level of control. The detected disorders are significantly exacerbated after a combination of acute blood loss and ischemia-reperfusion of the limb. The use of Carbacetam in comparison with animals without correction is accompanied by a significant decrease in the content of MMM254-280 fractions in the serum after 7 and 14 days of the experiment.