Coagulation Parameters in Post-Covid-19 Condition in Relation to Various Titers of Anti-SARS-CoV-2 IgG in Blood Plasma.

Background and Objectives: Post-COVID-19 condition is thought to affect 10-20% of people at least 3 months after a diagnosis of COVID-19 and two months of symptoms. Post-COVID-19 condition presents itself with many clinical effects with varying degrees of severity ranging from a mild cough to a life-threatening coagulopathy. Our study aimed to identify a relationship between the titers of anti-SARS-CoV-2 IgG and anticoagulation parameters: antithrombin III (ATIII), protein C (PC) and thrombomodulin (TM). Materials and Methods: Blood plasma was collected from healthy donors aged 25-45 who had recovered from COVID-19 3-6 months ago and their titers of anti-SARS-CoV-2 IgG and ATIII, PC, and TM were measured. Results: We found that concentrations and activities of key anticoagulation parameters (ATIII, PC, and TM) measured in donor plasma during the post-COVID-19 varied in relation to the titers of anti-SARS-CoV-2 IgG. Conclusion: While we identified a dysfunction of anticoagulation parameters in patients with post-COVID-19, we aim to explore the subpopulation antibody IgG fraction directly using in vivo and in vitro experiments with the possibility to contribute to the development of treatment options for post-COVID-19 conditions.

CHANGES IN THE SENSORIMOTOR CORTEX OF THE RAT BRAIN UNDER THE MODELING OF HEMORRHAGIC STROKE.

OBJECTIVE: The aim: To assess the structural and metabolic changes in the sensorimotor cortex of the rat brain under conditions of hemorrhagic stroke. PATIENTS AND METHODS: Materials and methods: The experiment was carried out on rats of the control and experimental groups with a model of hemorrhagic stroke. We used histological, electron microscopic, biochemical methods and biological markers. RESULTS: Results: In the sensorimotor cortex of the ipsilateral cerebral hemisphere of rats under conditions of hemorrhagic stroke, cerebral edema and progression of neurodegenerative changes were observed; an increase in the size of mitochondria, which is caused by edema of their matrix; activation of lipid peroxidation processes and a decrease in the activity of enzymes of the antioxidant system, a decrease in the level of apoptosis markers and inhibition of ERK1/2 expression. The study of DNA fragmentation in the cerebral cortex revealed a significant number of manifestations of necrosis and an insignificant number of cells in a state of apoptosis. CONCLUSION: Conclusions: after modelling a hemorrhagic stroke in the right hemisphere of the brain, perivascular and pericellular edema of the energy apparatus, cell death by necrosis and apoptosis, and activation of lipid peroxidation processes were established as well as a decrease in the activity of enzymes of the antioxidant system.

PROTEOLYTIC ACTIVITY IN THE HEART OF RATS WITH HYPERHOMOCYSTEINEMIA.

OBJECTIVE: The aim: To investigate the distribution of proteolytic activity and cytokine profile in the heart of rats with hyperhomocysteinemia. PATIENTS AND METHODS: Materials and methods: A total of 60 albino non-linear male rats was used in the study. Hyperhomocysteinemia was induced by intragastric administration of DL-homocysteine thiolactone. Total proteolytic activity was measured using casein as a substrate. To determine the activity of metal-dependent and serine proteases, ethylenediaminetetraacetic acid, and phenylmethylsulfonyl fluoride were used. The level of matrix metalloproteinases, tissue inhibitor of metalloproteinases-1, and cytokines was studied by enzyme-linked immunosorbent assay. RESULTS: Results: It was found an increase in the total proteolytic activity in the heart of young, adult, and old animals. In addition, the redistribution of proteolytic activity was revealed - the portion of metal-dependent enzymes increased in all groups while the percentage of serine proteases decreased in the old animals with hyperhomocysteinemia. The state of mild inflammation, evidenced by the increased level of some pro-inflammatory cytokines, was found in the heart of young and old animals with hyperhomocysteinemia. CONCLUSION: Conclusions: The pathogenesis of hyperhomocysteinemia is accompanied by a change in the proteolytic activity in the heart as well as a change in the cytokine profile.

CARDIOVASCULAR SYSTEM OF THE MATURE RATS WITH CONGENITAL HYPOTHYROIDISM AND ARTERIAL HYPERTENSION.

OBJECTIVE: The aim: Studying changes in the ultrastructure of blood circulatory capillaries of the myocardium of mature rats with hypothyroidism and arterial hypertension. PATIENTS AND METHODS: Materials and methods: Experiments were conducted on (240 days) 10 ISIAH (inherited stress-induced arterial hypertension) line rats with AH (arterial hypertension), 10 Wistar line rats with congenital hypothyroidism and 10 intact animals. Arterial pressure was measured, and the development of hypothyroidism was controlled by the immune enzyme method. The study of the left ventricle myocardium of the rat heart was carried out by electron microscopic and morphometric studies. RESULTS: Results: In in rats with AH the following changes were observed in the blood capillaries of the myocardium: decrease in the number of capillaries; disturbance of blood circulation; the number of organelles of the biosynthetic plan and structures involved in the transendothelial transfer of substances decreased in endothelial cells; lysis and edema of the latter; mucinous perivascular edema, confirmed by the accumulation of fine-fibrillar structures, collagen fibers, cellular detritus. By the same term, in the group with congenital hypothyroidism, dystrophic-destructive changes in the blood capillaries of the myocardium acquired the highest degree, which resulted in a decrease in their number due to destruction. Ultrastructure of the biosynthetic plan organelles and structures of the transendothelial transfer of substances were in decompensated state. CONCLUSION: Conclusions: The rats (in 240 days) with AH and congenital hypothyroidism express breakdown of compensatory processes in the capillaries of the myocardium. This is manifested by the further dilution of capillaries, the development of hypoxic state in them as well as mucinous edema of interstitium, the decrease of activity of biosynthetic and transport processes.

INFLUENCE OF COMBINED PHARMACOTHERAPY ON CARDIO-VASCULAR SYSTEM OF ARTERIAL HYPERTENSION (IN EXPERIMENT).

OBJECTIVE: The aim: Study of the patterns of structural changes in the left ventricular myocardial capillaries of rats with arterial hypertension with combined pharmacotherapy with Bisoprolol and Thiotriazolinum. PATIENTS AND METHODS: Materials and methods: Experiments were conducted on 30 line rats with congenital stress-induced arterial hypertension: 10 animals without treatment and 10 animals with treatment. Pharmacological correction of spontaneous arterial hypertension was performed with 20 mg / kg of Bisoprolol and 50 mg / kg of Thiotriazolinum per os once a day. Pharmacotherapy began at 5 months of age, that is, at a time when compensated heart failure was formed in rats with arterial hypertension. Animals were withdrawn from the experiment 100 days after the start of the correction. Control was provided by intact animals (10 rats) of the corresponding age. While extracted from the experiment rats of all experimental groups had their arterial pressure measured using a plethysmograph, electron microscopic examination of the left ventricular myocardium and morphometric study of volumetric and quantitative densities, cross-section area and form factor of micropinocytotic vesicles were conducted. RESULTS: Results: In rats with arterial hypertension after application of Bisoprolol and Thiotriazolinum, arterial pressure significantly decreases in experimental rats compared to animals without correction. The number of capillaries in the myocardium after pharmacotherapy increases up to control values, which shows their reparation. In most endothelial cells, organelles retain their integrity and presence that are characteristic of intact rats. The well-expressed processes of transcytosis are shown by the statistical similarity of the quantitative density and the size of the micropinocytotic vesicles in the endothelial cells of the myocardium capillaries of compared experimental animals. CONCLUSION: Conclusions: In rats with arterial hypertension, the combination of Bisoprolol and Thiotriazolinum prevents the decrease in the number of capillaries in the myocardium of the left ventricle, promotes the preservation of the ultrastructure of their endothelial cells and maintains the processes of transedothelial transfer of substances at the level of intact animals.

Morphofunctional status of cardio-vascular system of rats with arterial hypertension.

OBJECTIVE: The aim: Was to clarify the general patterns of structural changes in the left ventricular myocardial capillaries in rats with spontaneous arterial hypertension. PATIENTS AND METHODS: Materials and methods: Experiments were conducted on 50 ISIAH (inherited stress-induced arterial hypertension) line rats with arterial hypertension: juvenile young (45-day) and sexually mature (100-day) rats, as well as intact animals of the corresponding age. While extracted from the experiment rats of all experimental groups had their arterial pressure measured using a plethysmograph. Electron microscopic examination of the left ventricular myocardium and morphometric study of volumetric and quantitative densities, cross-section area, and form factor of micropinocytotic vesicles were conducted. RESULTS: Results: In sexually mature rats with arterial hypertension, a high level of pressure is maintained. In 45-day-old rats with arterial hypertension in endothelial cells of myocardial blood capillaries there is a hyperactivation of biosynthetic processes (euchromatic nucleus, large-sized mitochondria, ER canals, Golgi complex), which may be a manifestation of reactive processes in response to a non-stable increase in arterial pressure. In the 100-day rats with arterial hypertension, the mosaic of the ultrastructure of the myocardium blood vessels is preserved, but destructively-dystrophic changes become more expressive and involve not only the organelles but also the integrity of the endothelial cell itself. Destructively-dystrophic processes in rat capillaries are accompanied by compensatory and adaptive ones. This is manifested by activation of the transport of substances, both transendothelial and paracellular, and quantitative density of micropinocytotic vesicles increases statistically significantly. CONCLUSION: Conclusions: In myocardial capillaries of young (45-day) arterial hypertension rats, compensatory and adaptive changes are manifested by activation of biosynthetic processes in endothelial cells following a slight increase in micropinocytotic vesicles quantitative density and signs of destructive-dystrophic processes (minor edema and lysis of endothelial cell cytoplasm). In sexually mature (100-day) arterial hypertension rats in the blood capillaries of the myocardium, the destructive-degenerative changes increase is accompanied by preservation of signs of compensatory processes. Reducing the number of capillaries is offset by an increase in the number of micropinocytotic vesicles.

The influence of drug treatment on cardio-vascular system of the rats with congenital hypothyroidism.

OBJECTIVE: Introduction: In the treatment of hypothyroidism substitution therapy with L-thyroxine is used, it is also advisable to use the metabolites with membrane-stabilizing properties that normalize the metabolism in the body, for example, calcitonin, which significantly reduces the depth of the dystrophic phenomena in the myocardium. The aim was to study the patterns of structural changes in the left ventricular myocardial capillaries of rats with congenital hypothyroidism in combinative drug therapy with L-thyroxine and calcitonin. PATIENTS AND METHODS: Materials and methods: 30 white Wistar line rats were used as experimental animals: 10 with treatment and 10 without as well as control - 10 intact Wistar line rats of the same age. Mercazolil was used to inhibit thyroid gland in order to model congenital hypothyroidism. After birth, the rats received L-thyroxine at a dose of 10 µg / kg per os daily, calcitonin at a dose of 1.0 MU / kg per day intramuscularly, then with mother's milk, later by themselves for 100 days. The arterial pressure was measured in all experimental groups during extraction from the experiment by plethysmograph, their left ventricular myocardium was examined under electron microscope and micropinocytotic vesicles in their cells were studied morphometrically. RESULTS: Results: In rats with congenital hypothyroidism, for which L-thyroxin drug in combination with calcitonin was used as a substitution therapy, after pharmacological correction, in general there is no pronounced heteromorphism of the ultrastructure of the left ventricular myocardial blood capillaries, which was characteristic for animals without pharmacological correction. The analysis showed normalization of the content of free thyroxine in blood plasma and blood pressure of rats with congenital hypothyroidism after complex substitution therapy. CONCLUSION: Conclusions: In rats with congenital hypothyroidism, which received L-thyroxine and calcitonin at birth, the myocardium capillaries generally remain intact and have morphological and functional characteristics similar to intact animals, which is the theoretical basis for the need for calcitonin to be used in substitution therapy in hypothyroidism.

Morphofunctional status of cardio-vascular system of rats with congenital hypothyreosis.

OBJECTIVE: Introduction: There is an increasing number of cases of congenital hypothyroidism. One of the most common complications of hypothyroidism is damage to the cardiovascular system, which in 30-50% of patients leads to the development of arterial hypertension. The aim: studying the features of the ultrastructure of myocardial capillaries in mature rats with hypothyroidism. PATIENTS AND METHODS: Materials and methods: Experiments were conducted on 40Wistar line rats with congenital hypothyroidism: juvenile young (45-day) and sexually mature (100-day) rats, as well as intact animals of the corresponding age. While extracted from the experiment rats of all experimental groups had their arterial pressure measured using a plethysmograph while the development of hypothyroidism was controlled by the immune-enzymatic method. Electron microscopic examination of the left ventricular myocardium and morphometric study of volumetric and quantitative densities, cross-section area, and form factor of micropinocytotic vesicles were conducted. RESULTS: Results: In the sexually mature rats with congenital hypothyroidism the quantitative density of the capillaries in the myocardium decreases. Activation of transcytosis is accompanied by significant violations of vesiculation. Some of the endothelial cells of experimental animals contain a moderate amount of transport vesicles, while others are overfilled with these structures and desquamate into the lumen. In older rats with congenital hypothyroidism there is a further dilution of capillaries, the development of hypoxic state in them, mucinous edema of interstitial space, decrease of biosynthetic and transport processes activity. CONCLUSION: Conclusions: In young (45-day) rats with congenital hypothyroidism the direction and expressiveness of compensatory processes is to enhance the transcytosis processes. Dystrophicdestructive changes are manifested by apoptosis in some endothelial cells, decrease in the number of biosynthetic organelles, lysis and edema of their cytoplasm. In sexually mature (100-day) rats with congenital hypothyroidism destructively-dystrophic processes in the blood capillaries of the myocardium are approximately balanced with compensatory-adaptive.

Experimental study of the liver ultrastructure in the presence of infection with herpes simplex virus 1.

OBJECTIVE: Introduction: Understanding of HSV-1liver infection pathogenesis is of great scientific, social and economic significance, since this is one of the main latent infections in population. However reactivation of this infection remains understudied. The aim: This experimental research aimed at studying the ultrastructure changes occurring in the liver in the presence of HSV-1infection. PATIENTS AND METHODS: Materials and methods: Experiments were conducted on 12 BALB/c line mice weighing 18-20 g. They were divided into 2 groups: experimental, and control. Experimental animals were infected with the attenuated HSV-1. On day 40 the animals were withdrawn from the experiment by decapitation. Liver fragments were excised and studied ultramicroscopically. RESULTS: Results: Liver disorders were represented by the focal damage of hepatic lobuli cells. Ultrastructure changes were found both in the microvascular endothelium and hepatocytes. The vascular disorders included swelling of endotheliocytes, their demise and desquamation into the lumen, disruption of the basal lamina integrity and diapedesis of blood cells into the subendothelial space. Finding virions in the endotheliocytes allowed to explain the possible pathway of the infection into the interstitium and hepatocytes via systemic circulation from the primary source of infection. Electron microscopy has not revealed any virions in hepatocytes, with only the following changes: significant cytosole density of the osmiophylic granules, lisosomes and lamellar bodies found. These were considered to be the consequence of the infectious process. Findings of the experimental study enable understanding of the causal relationship between the acute infection and liver damage. CONCLUSION: Conclusions: Ultrastructure changes in the liver of mice infected with HSV-1 were focal, and more rarely diffuse in nature. Non-specific cytopathological changes (swelling of the cytoplasm and reduction of the endoplasmatic reticulum, and mitochondria) were found both in the endotheliocytes of the sinusoid capillaries and hepatocytes. Endotheliocytes of the sinusoid liver capillaries in mice infected with HSV-1 lose their barrier function, which leads to direct and indirect damage of hepatocytes and development of dystrophic changes in the liver.

[Ultramicroscopic features of cells and vessels of the spleen (experimental study)].

OBJECTIVE: Introduction: The spleen is a multifunctional odd organ, which is a biological filter, but its structure in experimental animals is not fully understood. PATIENTS AND METHODS: Materials and methods: The ultramicroscopic structure of the cells and vessels of the spleen of amphibian animals (Pelophylax lessonae С. - the pool frog and Pelophylax ridibundus Р. - the marsh frog) are elucidated in the research. RESULTS: Results: The cytopupulation of the spleen is formed by lymphocytes, neutrophils, erythrocytes, macrophages, plasma cells etcetera. A feature of the cellular composition of the parenchyma of the spleen of the frogs is a significant number of neutrophilic eosinophils. The organelles, which situated in the cell cytoplasm, are characteristic of somatic cells. The endothelial lining and the structure of the capillary basement membrane, which has the form of a plate, formed from 8-12 endothelial cells, grouped in complexes and adjoining contacts in adjacent zones, are experimentally substantiated by the somatic type of vascular endothelium of the microcirculatory channel. Such cells are contained on the basal membrane, surrounded by a dense interlacing of collagen, elastic fibers and the main substance, and the sites of endothelial cell adhesion have the uneven contours. The nuclei of the endothelial cells are surrounded by a cytoplasm on all sides, which reveals foamy cysts. CONCLUSION: Conclusions: The results obtained are of great interest for evolutionary immunomorphology, the extrapolation of which improves understanding of microcirculation, transport of substances in the spleen, and mechanisms for formation of the immune response.

The influence of lead nanoparticles on the morpho-functional changes of rat liver during the postexposure period.

Lead as any heavy metals may be found in soil, water, air, and is used in everyday life. Once in the body, it causes toxic effect, making the liver, which is one of the main organs of detoxification, suffer. Recently, the study of the action of not only ionic forms of lead, but also its nanoparticles, has become topical. The study aims at determining changes in the liver of rats and biochemical changes in their blood both at late term of exposure to nanoparticles of lead compounds and in the post-exposure period. The study was performed on 120 male rats of Wistar line, which were divided into two series, each series containing four groups. The first and the second groups of animals were intraperitoneally injected with colloidal solution of nanoparticles of lead sulfide of 10 and 30 nm in size, and the third group were intraperitoneally injected with a solution of lead nitrate. The fourth group of animals served as control. In the first series, the investigated substances were administered 60 times within 12 weeks. In the second series, after 60-fold administration of the investigated substances, the exposure was discontibued and animals were observed for 6 weeks-overall duration of 18 weeks. Histological, morphometrical and biochemical methods were used. The body weight was reduced in the rats exposed to PbSnano1 at week 12 of experiment and in rats exposed to both PbSnano1 and Pb(NO3 )2 in the second series. Absolute liver weight increased at week 12 of experiment in all experimental groups. In the second series this value almost reached that of the control level. Relative liver weight in the animals of all experimental groups was higher than that in the control at week 12 of experiment. In the second series this value remained higher in rats exposed to PbSnano1 . After 12 weeks of exposure dystrophic changes in the liver were found in all experimental groups. At week 6 after the exposure (the second series) destructive changes in the liver decreased. Total protein, albumin, glucose, total lipids, cholesterol, triglycerides content in blood serum corresponded with morphological data. The experiment has demonstrated that the 12 weeks long exposure to lead nanoparticles had harmful effect on the liver. Within the postexposure 6-weeks period structural changes in the liver and biochemical changes in blood serum decreased. Biochemical changes in blood serum corresponded to the morphological data. By many parameters PbSnano1 had more pronounced harmful effect. Toxicity of PbSnano2 and Pb(NO3 )2 were comparable.

The study of leukocyte phagocytic activity in the presence of herpetic infection and stroke.

OBJECTIVE: Inrtoduction: Post-stroke complications are one of the urgent and insufficiently resolved problems. According to different literature data 23% to 65% of patients suffer from the post-stroke development of an infectious process. Herpes simplex virus type 1 and 2 can also be etiological factors of stroke development, however their reactivation is seldom mentioned in clinical observations. The development of immune suppression is considered to be the cause of these complications. The aim: The current study aims at determining post-stroke changes in leukocyte component of the immunity and in the presence of concomitant herpetic infection as well as at finding changes in phagocytosis parameters during antiviral treatment. PATIENTS AND METHODS: Materials and methods: The experiments were carried out on mice of the Balb/с line. The animals were infected with the herpes simplex virus type I, and 30 days later hemorrhagic stroke was simulated by administering 0.1 ml of autoblood into the right hemisphere. Following the acute stroke some animals were given acyclovir, proteflazid or altabor. From the animals' blood leukocytes were obtained and phagocytic activity and production of reactive oxygen species of granulocytes and agranulocytes in relation to fluorescent E.coli bacteria were studied by flow cytometry. RESULTS: Results: The experiment revealed significant changes in the redistribution between two major types of leukocytes in mice with stroke (an increased number of agranulocytes by 19.9%) and decreased phagocytosis activity, in the animals infected with herpes simplex virus type І in particular. Ischemic brain damage had an immunosuppressive effect on blood leukocytes. For comparison a significant increase in phagocyte count in leukocytes was found in the case of viral infection. The use of drugs with antiviral effects did not affect the activity of granulocytes / agranulocytes. CONCLUSION: Conclusion: Stroke can be the cause of latent herpes virus infection reactivation and has essential negative effect on immune characteristics of leukocytes that remain unchanged with the use of antiviral agents.

Rat spinal ganglia in assessment of protective action of antioxidants: A morphological study.

BACKGROUND AND OBJECTIVE: Mercury pollution is one of the most pressing environmental problems. Therefore, the impact of mercury on human body, the nervous system in particular, remains topical. The aim of the study was to identify the morphological characteristics of neurons and neuroglia in spinal ganglia of rats receiving antioxidants in the presence of small doses of mercury (II) chloride. MATERIALS AND METHODS: A total of 100 white Wistar rats were divided into 5 series (10 groups), with 10 animals in each group. The first series comprised intact animals receiving saline solution instead of drugs administered in other series (control). In the second series 10 injections of mercury (II) chloride were performed (group of short-term neurointoxication) and 50 injections (group of long-term neurointoxication). In the third to the fifth series, the short- and long-term neurointoxication was followed by 10 daily injection of the drugs: unithiolum, thiotriazolinum and mildronate respectively. Spinal ganglia were obtained two weeks after the completion of drugs administration and studied microscopically and ultramicroscopically. RESULTS: Administration of thiotriazolinum, unithiolum and mildronate mitigated manifestations of toxic effects of mercury (II) chloride on spinal ganglia. Unithiolum and thiotriazolinum activated synthetic processes, while mildronate had a positive effect on restoration of cells metabolism. CONCLUSIONS: Morphological data show that unithiolum and thiotriazolinum action decreases toxic effects of mercury chloride and are similar. They demonstrate pronounced activation of synthetic processes in sensory neurons and satellite cells of spinal ganglia. Mildronate also restores cell ultrastructure and has more pronounced effect on their energetic processes and interaction between neurons and satellite cells.