Vol 146, No 6 (2014)

Regression analysis was performed to detect the relationship between the changes in the histological structures of the brain (neurons, glial cells and microvessels) and the degree of ethanol poisoning (according to the data of alcohol concentration in the blood and urine). The material was obtained from 40 bodies of men and women aged between 20- 81 years. It was shown that the reduction of the diameter of microvessels in the cerebral and cerebellar hemisphere cortex and its increase in the white matter, together with the increased number of blood vessels with the stasis, of astrocytes demonstrating cytoplasmic granularity and of various signs of brain edema, as well as the reduction in the number of processes of astrocytes and the stainability of red blood cells in the zone of stasis with glycine cresyl red, correlated with the root mean square ethanol concentration (RMSEC) in the blood and urine. These results give the opportunity to assess this index during the histological examination of the brain. On the contrary, knowing RMSEC, it is possible to evaluate the presence of certain histological changes in the brain.

An immunohistochemical method was used to study the effect of capsaicin treatment on morphometric and structural characteristics of neurons containing 28 kDa calbindin (CAB) in the posterior horns of the spinal cord (SC) gray matter T II segment and in the sensory ganglion of the second thoracic spinal nerve (SGSN) in Wistar rats (n=4). Capsaicin was administered to adult animals 3 times with an interval of 24 hrs, in a total dose of 125 mg/kg, the material was taken on the 14th day. The administration of capsaicin caused a decrease in the proportion of CABimmunopositive (CAB-IP) neurons in SGSN (by 60%) and in dorsal horn laminas I-II-III (by 8, 18 and 15%, respectively), while the average size of CAB-IP neurons increased due to intracellular edema. As a result of deafferentation, similar morphometric and structural changes of CAB-IP neurons developed in both SGSN and posterior horn of SC gray matter, which were manifested by the central chromatolysis, vacuolation of nucleus and cytoplasm indicative of hydropic dystrophy. The irreversibility of the changes observed in the neurons of SGSN and SC dorsal horn laminas I, II and V was supported by the observations of their nuclear deformation, lysis of nucleolus, reduction of the number of CAB-containing neurons, signs of neuronophagia with the formation of residual nodules in place of the dead cells.

In male Wistar rats (n=25) aged 3, 10, 20, 30 and 60 days, subjected to chemical deafferentation by a single injection of capsaicin at postnatal Day 2, localization, relative content and morphometric characteristics of neurons expressing NO synthase (NOS) were studied in caudal ganglion of vagus nerve (CGVN) and sensory ganglia of spinal nerves (SGSN) using immunohistochemical and morphometric methods. The control group consisted of rats (n=25) of the appropriate age. The results suggest that in the control group the proportion of NOS-immunopositive neurons in SGSN increased during the first 10 days of life and decreased between Day 30 and Day 60. In CGVN, the proportion of NOS-immunopositive neurons did not change significantly during the ontogenesis. In animals of the experimental group, the proportion of NOS-positive neurons decreased rapidly during the first 20 days of life. More pronounced decrease in the number of NOS-containing neurons was observed in SGSN as compared to that one in CGVN. The data obtained indicate deleterious effects of capsaicin on NOS-positive neurons, which confirms the role of NO in the mechanisms of nociception.

Neuropeptide Y (NPY)-containing fibers and intramural ganglionic neurons, as well as Y 1 and Y 2 receptors to NPY were detected in the myocardium of rats during postnatal ontogenesis using immunohistochemical methods. The data obtained suggest age-dependent changes of the NPY-mediated heart innervation. The density of NPY-positive fibers was the least in newborn animals and increased during first 20 days of life. Y 1 and Y 2 receptors were found in the heart of newborn and more adult rats. Y 1 receptors were observed in the coronary heart arteries, arterioles and cardiomyocytes from the moment of birth. The density of NPY Y 2 receptors was very low in the cardiac muscle of young (prior to the 2 nd month of age) and adult (up to 1 year-old) rats but substantially increased in aged animals. During the postnatal ontogenesis? all the intramural ganglionic neurons were Y 1-positive and Y 2-negative.

Gastrocnemius muscle of outbred female albino rats was studied from postnatal Day 14 to Day 180 in normal animals (control group, n=18), after neonatal chemical deafferentation (n=18), after neonatal chemical desympathization (n=18) and after combined neonatal chemical denervation (deafferentiation plus desympathization, n=18). Histochemical methods were used to demonstrate cholinesterase activity (substrate: thioacetic acid) and alkaline phosphatase (substrate: naphthol AS-BS phosphate). adaptation capacity and a tendency to earlier development of After combined chemical denervation compared with control a reduction in the activity of cholinesterase was observed, leveling of the early age elimination of neuromuscular synapses, lower proportion of complex enzymatic zones, and later formation of the vascular loops around the motor endings. The vascular network was characterized by a larger diameter and a predominantly planar orientation of the vessels. Developing «muscle fiber - neuromuscular junction - vessel net» system had low involutive processes.

Morphometric methods were used to examine the dynamics of the quantitative parameters of the surviving portion of the adrenal medulla (AM) in Wistar rats (n=41) for 2 months after its laser damage. On paraffin sections of AM stained using L. H. Honore method, the volume fractions and the average cross-sectional areas of adrenalocytes and noradrenalocytes as well as the volume fractions of blood vessels, stroma and neurons were determined. The relative volume of the blood vessels reached its maximum value at Day 7, and that of the chromaffinocytes - at Day 28 of the experiment. The volume fractions of the stroma and neurons demonstrated no significant changes during the experiment. The original method was used to calculate the real decrease and increase in the volume of glandular parenchyma of the surviving portion of AM. The real decline of chromaffinocyte volume reached a maximum of 1.5% at Day 7, and the maximum real growth of 4.7% was observed at Day 28 (the growth rate amounted to 1-2.5% per week). Until Day 28 chromaffinocyte hyperplasia dominated in the regenerative processes, while on Day 28 the hypertrophy was also found to significantly contribute to it. By Day 56 the surviving zone of AM was normalized.

The state of the liver was studied in 13 puppies with experimental aortic coarctation, 12 animals with pulmonary trunk stenosis and 10 control (intact) dogs of the appropriate age. Histological, morphometric methods and immunohistochemical reaction with the monoclonal antibodies against α-SMA were used. Studies have shown that aortic coarctation led to hypotonia and atrophy of the walls of the arteries, branches of the portal vein and veins related to the hepatic vein system. On the contrary, in pulmonary trunk stenosis the thickening of the walls of branches of the hepatic veins and the development of veno-arterial and veno-venous reactions were observed. Despite the significant differences in hemodynamic disorders in these defects, an increase in the number and degree of development of the adaptational muscular structures was found in the afferent vascular bed of the liver. In the efferent vascular bed, in coarctation, the relaxation and the atrophy of muscle cushions in the veins related to the hepatic vein system was found, which was accompanied by a decrease in blood storage in the liver. In pulmonary trunk stenosis, to prevent venous congestion, the hypertrophy of the muscular cushions occured in these vessels. After some time, in both defects, hepatic blood vessels demonstrated angiosclerosis; degenerative changes developed in the parenchyma and sclerosis - in the stroma.

Using a complex methodology of in vivo monitoring of the motor activity of the ciliary apparatus in combination with histological and morphometric methods, the epithelial lining of the trachea 110 Wistar rats was studied from birth until old age (24 months). In newborn animals, tracheal epithelium was characterized by the minimal content of the ciliated cells (18%), the maximum frequency of the ciliary beat (FCB) -15 Hz and the lack of directional movement of mucus. Within the 1st month of life, the most intensive growth in the number of the ciliated cells and their morphometric parameters (the height of the cell, the length of the cilia) was marked together with the formation of the pseudostratified structure of the epithelium. By the end of this period morphologically identifiable goblet cells first appeared. The emergence of mucociliary transport (0.03 mm/s) was registered since Day 14 after birth. Tracheal epithelium reached structural and functional maturity by the 3rd month of postnatal development, at this time, the velocity of mucus movement (VMM) increased to 0.1 mm/s, FCB at the age of 6-12 months was equal to 11.3-11.7 Hz. In the aged (24 month old) animals FCB increased to 12.9 Hz, while the morphometric parameters of the ciliated cells, the proportion of goblet cells and VMM reached their maximum values. At the same time, the number of the basal (cambial) cells in the epithelial layer was reduced, while the number of dying cells was increased. The results of the study indicate a close relationship between processes of structural and functional differentiation of the ciliated epithelium of the respiratory tract ensuring the effective operation of mucociliary apparatus throughout the life.

Reversible hemodynamic model of pulmonary trunk stenosis was created in experiments on 34 puppies, 12 of which were later subjected to corrective surgery. Animals of both groups were followed up for 6-12 months. 12 dogs of respective age were used as a control group. The kidneys of animals of all series of experiments were studied using a complex of histological and morphometric methods. It was shown that the constriction of the pulmonary trunk lead to adaptive and pathological remodeling of renal arteries, veins and glomeruli. The former became apparent as the hypertrophic-hyperplastic changes in renal vascular smooth muscle with thickening of the vessel walls, narrowing of the arterial lumen and the remodeling of some of them into the closing type. The latter were manifested by vascular sclerosis of arterial and venous vessels, glomerular collapse and sclerosis. Significant changes were detected in renal tubules and stroma: tubules became atrophic, while stroma underwent sclerosis. All these changes reflect structural remodeling of the kidneys. Elimination of experimental defect triggered the regeneration mechanism resulting in the «reversal» of some changes that developed earlier. During this process, the complete recovery of all renal tissue components was not achieved.

The aim of the study was to establish the possibility of assessment of the state of peripheral nerve based on acetylcholinesterase (AChE) activity. In a series of experiments on rats, the method for demonstration the of AChE activity on the sections of the peripheral nerves was modified (priority reference № 2013157128 of 23.12.13) and it was shown that the reaction intensity depended upon the state of the nerve fibers. During the atrophic processes, the enzyme activity decreased, while it was dramatically increased during fiber sprouting. It was shown that by using the intensity of AChE reaction as a criterion, the state of the nerve could be evaluated within 1-2 hours, which may be used later during the reconstructive operations (intraoperatively).