[Immunolocalization of hemeoxygenase-2 in neurons of the human vasomotor center in arterial hypertension].

We studied the immunolocalization of hemeoxygenase-2 in neurons of the medulla oblongata in men (n=8), aged 18-44 years, who died from causes unrelated to the injury of the central nervous system and in people with the lifetime diagnosis of hypertension (n=6). It has been found that neurons with enzyme positive reaction are present in all parts of the medulla oblongata with concentrations ranging from 0.5 to 13.7% of the total number of cells. The high proportion of small neurons with the high or moderate density of deposits was found in the sensory nuclei. Large cells of the motor nuclei often exhibit the negative or low intensity of the enzymatic reaction. In arterial hypertension, a decrease in the proportion AH NO-positive neurons and the average optical density of the reaction product was noted. The reduction was seen in most affected neurons in the rostral part of the solitary tract nucleus and the lateral reticular nucleus. In the motor nuclei and in the dorsal nucleus of the vagus nerve, these parameters decreased as well although the reduction was not as great as observed in the sensory nuclei.

[Histochemical and immunohistochemical localization of choline acetyltransferase in the nuclei oblongata rat brain].

ChAT-positive neurons in the nuclei of the medulla oblongata of Wistar rats have been studied with the use of histochemical and immunohistochemical methods. We have found that the topography and the number of cholinergic neurons at the projection nuclei studied largely depend on the method of detection of these neurons. Histochemical method always revealed more neurons than immunohistochemistry. Such a feature of ChAT-positive neurons detection was clearly seen among the majority of nuclei in the medial region and in some of the nuclei in the lateral region of the medulla oblongata. The number of immunoreactive cells in the nuclei varied from 17 to 26%, whereas the histochemical reaction determined 1.5-3 times more neurons in the same nuclei. ChAT-positive cells in the nuclei of the back seam were detected mainly by a histochemical method.

[Heme oxygenase-2 neurons brain and spinal cord of human].

Immune localization of heme oxygenase-2 in neurons of some nuclei of the spinal cord and brain stem in 6 men 18-44 years old who died from causes unrelated to injury of central nervous system was studied. Neurons with positive reaction are determined for all studied regions of the brain where their contents in various nuclei ranging from 0.5 to 16% of the total number of cells detected by methylene blue. In all the sensory nuclei there is a high proportion of small neurons with a high or moderate density of reaction produce deposits. Large cells of motor nuclei often exhibit negative or low intensive enzyme reaction.

[Immunocytochemical characterization H2S-positive neurons in the nuclei of bulbar cardiovascular center in the development of renovascular hypertension].

Immunocytochemical method in Wistar rats in 6 nuclei of the medulla oblongata, related to the cardiovascular center, identified nerve cells (H2S-neurons) with a positive reaction to cystathionine beta-synthase (CBS)--an enzyme involved in the synthesis of hydrogen sulfide in the brain. Found that in rats with renovascular hypertension only after 6 weeks of hypertension in the neurons of the reticular nuclei observed a significant reduction of CBS, while the number of H2S-positive cells in this period does not change significantly. Only at week 12 renovascular hypertension notes marked reduction in the proportion of positive H2S-neurons in the reticular nuclei of most of the medial zone, although further reduction in the intensity of the enzymatic reaction in the neurons were observed. Between 14-16 weeks of hypertension to further reduce the proportion of neurons and H2S-content of the enzyme is not observed. In a single nucleus and reticular lateral nucleus, which receive afferent impulses in the development of renovascular hypertension specify the minimum change of the indicators.

[H2S-positive neurons in some nuclei of cardiovascular center of rat brain].

The aims of this study were to investigate the presence of H2S-positive (H2S+) neurons and to study their distribution of in the nuclei of the medulla oblongata and the pons which are included in the cardiovascular center. Using immunocytochemical method to detect cystathionine beta-synthase, the presence of H2S+ neurons was demonstrated in the nuclei studied of Wistar rats (n = 16). More intense the reaction was detected in the large cells of motor nuclei, whereas the small cells of sensory nuclei usually had low-to-moderate enzyme activity. The data obtained provide a considerable amount of new information regarding the organization of brain nerve centers, including those controlling the cardiovascular system.

[Immunohistochemical study of H(2)S-positive neurons in some brain structures of people with arterial hypertension].

The post-mortem study included 8 healthy men, aged 18-44 years, and 27 men with the life-time diagnosis of hypertension, stage I-III. Using immunocytochemical method, we studied topography, quantitative distribution and enzyme activity in H2S-positive neurons of some nuclei of the medulla oblongata and pons that are part of the cardiovascular center. Changes in cystathionine-beta-synthase activity and in the number of immunopositive cells were found in arterial hypertension. The intensity of these parameters was correlated with the severity of disease. It has been proposed that changes in morphometric parameters of H2S-positive neurons inside the nuclei reflect the character of between-nuclei interactions as well as the organization of the cardiovascular center in whole.

[Cystathionine beta-synthase in the structural elements of the human brain and spinal cord].

Investigated by immunohistochemistry the presence and distribution of CBS in the nuclei of the spinal cord and brainstem in 8 mens of 18-44 years old, died from mechanical injury is not related to the central nervous system damage. Established that the CBS-neurons are revealed in all parts of stem and spinal cord, but their content in the studied cores, ranging from 0.9 to 17 %. Observed a definite relationship between the activity of CBS and the quantity of neurons. Large cells of motor nuclei often have a high and very high intensity of the reaction. In the sensory nuclei of a high proportion of small neurons with low activity of the enzyme.

[The comparative characteristic seratonergic neurons in some nucleus of the oblong brain of the rat].

Immunohistochemistry in Wistar rats identified serotonergic neurons in 8 cores of the medulla oblongata belonging to the so-called "bulbar vasomotor center". Using morphometry revealed that the proportion of serotonergic neurons located in the projection of the cores studied ranged from 17 to 26 %, and value of the index rised to 34-40 % only in the cores of the back seam. Single immunopositive cells that can perform the integrative function in the regulation ofhemodynamics were detected between the cores as well as between the cores and pathways.

[Serotonin- and nitroxidergic neurons of rat medulla oblongata].

The purpose of the present work was topochemical mapping of serotonin- and nitroxidergic neurons in medulla oblongata of 12 Wistar rats in eight nuclei envolved in so-called "bulbar vasomotor to the center". It was found that a portion of serotoninergic neurons lying in a projection of the investigated nuclei, was equal, on the average, to 12-15%, and those in the nuclei of posterior raphe group increased up to 31-43%. Nitroxidergic neurons were distributed more uniformly, and their portion in different nuclei varied from 19 to 49%. 2-6% of neurons were located between the nuclei, and between the nuclei and conducting pathways; these cells presumably carry out integration functions in hemodynamics regulation.

[Receptor system of brain vessels in arterial hypertension].

We conducted a post-mortem study of the receptor system of brain vessels in patients with arterial hypertension (AG), stages I-III, and healthy people using impregnation and a method of measuring NADPH-diaphorase. The receptor system of arterial wall is represented by treelike and glomerular structures with moderate enzyme activity. Reactive and destructive disturbances of the receptor system that were distinctly seen on histochemical preparations were found in brain arteries of AG patients. The intensity of changes in the nervous apparatus of brain vessels was closely related to the artery caliber, their localization, disease duration and severity, with the most early and deep changes in the afferent fibers and receptors of the pia mater arteries. In intracerebral arteries, reactive changes in receptor structures were found in long-term hypertension and destructive changes were found in AG, stages II-III.

[Age-related changes in nitroxidergic neurons in some nuclei of rat medulla oblongata].

The paper presents a comparative study of NO neurons in the solitary tract nucleus, giant-cell, and lateral reticular nuclei in rats at 4, 7, 10, 14, 30, 45, and 60 days old and 3, 6, 12, 18, 24 months old. We determine the active quantitative and qualitative changes that occur in NO-positive neurons in the studied nuclei during the course of postnatal development. A low level of enzyme activity is observed on the first day; it reaches a peak level around the first-third month, then slowly declines. The size and number of nitroxidergic neurons increases, while the relative cell density decreases until the third month of life. We reveal local differences in the ontogenetic development of NO neurons in the studied nuclei. Solitary tract neurons have the highest rate of development, while NO neurons of old animals undergo early and extreme changes as compared to other studied nuclei of rat medulla oblongata.

[Nitricoxideergic neurons of the human bulbar vasomotor center in arterial hypertension].

The distribution of nitricoxideergic neurons and activity of neuronal NO-synthase (nNOS) was studied in some nuclei of the medulla oblongata in patients with the lifetime diagnosis of arterial hypertension, stages I-III (AG I-III). In AG I, the significant decrease of nNOS activity was observed in most nuclei, although the relative content of NO-neurons was not changed. The marked changes of these parameters were noted in the nucleus of solitary tract compared to those of the reticular formation. In AG II, a portion NO-neurons in the nuclei was markedly reduced, however the following decrease of nNOS activity was not found. In AG III, there was the augmentation of nNOS activity and the subtle decrease in the portion of NO-neurons. Therefore, in AG, there is the decrease of quantitative parameters that characterize the state of the oxide-ergic system. This is one of the causes of the hyperactivation of sympathetic nervous system and elevation of arterial pressure. The exception is the dorsal nucleus of the vagus nerve, in which the relatively larger number of NO-neurons and higher nNOS activity were found, that may be related with the cholinergic mechanism of compensatory activation of the nitric oxide-ergic system.

[Nitric oxide in the mechanisms of afferent innervations of arteries of the brain].

Structural elements of afferent innervations of brain arteries in rats such as receptors and nervous fibers, neurons of ganglia jugularis unit and the nucleus of a single way were investigated with the help of histochemical and electron cytochemical methods. The presence of three types of receptors and afferent fibers has been established in vessels. Neurons with positive reaction to NADPH-diaphorase have been allocated in the ganglia jugularis unit and the nucleus of a single way (17.4 and 24.5% accordingly).

[Efferent innervation of the arteries of human leptomeninx in arterial hypertension].

Structure of the efferent nerve plexuses (adrenergic, acetylcholinestherase- and cholinacetyltranspherase-positive, NO-dependent), was studied in the arteries of human leptomeninx with different diameters. Material was obtained from the corpses of the healthy people and of the patients with initial stages of arterial hypertension (AH). It was shown that the concentrations of cholinergic and adrenergic nerve fibers and varicosities in axon terminal part, innervating the arteries with the diameters ranging from 450 till 100 microm, were not significantly different. In these arteries, NO-ergic plexuses were also detected. In patients with AH, regardless the arterial diameters, the significant increase (up to 15-20%) of adrenergic nerve fiber and varicosity concentrations was found. The changes in cholinergic nerve fiber concentration were found to depend on the vessel diameter: the significant decrease of these parameter was observed only in arteries with the diameter of 100-200 microm. No significant changes in nerve plexus concentration was noticed in the arteries with greater or smaller diameter. In NO-ergic neural conductors, the enzyme activity decreased only in the large arteries, and remained almost unchanged in the small vascular branches. The changes in the vasomotor innervation described in AH, are interpreted as a vasomotor innervation dysfunction of the leptomeninx arteries that may result in the hemodynamic disturbances.

[Nitroxidergic neurons in some nuclei of medulla oblongata in human and rat].

Seven nuclei of medulla oblongata bearing a relation to human and rat bulbar vasomotor center and in which four types of neurons were distinguished by intensity of reaction to NADPH-diaphorase were analyzed. The analysis has shown that majority of human and rat nuclei have similar pattern on nitroxid-positive neuron distribution: neurons with high NO-synthase activity predominate in vasomotor nuclei, and neurons with low one predominate in sensitive nuclei.

[Vasomotor innervation of the human pia mater in arterial hypertension].

Materials of forensic-medical autopsies of men, aged 18-44 year, who died from trauma, have been studied. Arteries of the pia mater have been taken from 26 men with arterial hypertension (AH), stages I-III, and 12 men without AH. Histochemical methods have been used to study the structure of nerve plexus of medial brain artery branches of different diameters. In AH, there were changes of the nerve plexus structure in brain arterial branches, the intensity of which was related to the vessel diameter and disease stage; the decrease of density of periadventicial adrenergic nerve plexus compared to controls, which was most intensive in AH, stage I, and in the vessels of smaller size. The changes in concentration of distinctly seen bulges (varicosities) were more significant compared to those of nervous fibers. The effectiveness of nitrooxidergic innervation was also decreased in large vessels and its structure did not visibly change in small arterial branches. In conclusion, there is a formation of vasomotor innervation dysfunction in AH.

[Nitroxidergic nerve fibers of intracerabral blood vessels].

Methods of light and electron microscopic histochemistry were applied to study the structure and distribution of NADPH-diaphorase-positive neurons and processes in the parietal area of rat cerebral cortex. It was found that the most of the neurons displayed close connections with the intracerebral vessels. In the cerebral cortex, the smallest distance between the axonal plasma membrane and smooth muscle cells of the intracerebral arteries was found to be no less than 0.3-0.5 microm. Neuronal cell bodies were located in the functionally important areas of the vessels (in the areas of lateral trunk branching and in arteriolar sources), while their processes accompanied the vessels, tightly embracing them with their branches. Quite often, the neurons, the dendrites of which make contacts with the bodies or processes of over- or underlying neurons, sent their nerve fibers to the arteries, veins and capillaries. Thus, nitroxidergic neurons or their groups may control the blood flow in the different areas of vascular bed, performing the functions of the local nerve center.

[Ultrastructure of the intima of human pial arteries in arterial hypertension].

Ultrastructure of the intima of human pial arteries obtained from 5 male cadavers of practically healthy individuals and from 8 cadavers of the patients with the intravitally diagnosed grade I arterial hypertension (AH) was studied by scanning and transmission electron microscopy. AH was found to be associated with the remodeling of the intimal structural elements in the pial arteries. In most arteries, the changes were detected in the microrelief of the luminal surface and in the permeability of the vascular endothelial lining and of the subendothelial layer. During this remodeling, some endothelial cells were found in the state of structural and functional adaptation to the elevated arterial pressure, while the others were undergoing the dystrophic changes. The latter include the cells containing lipid inclusions, as well as the endothelial cells presumably in the state of apoptosis. The destruction of the intercellular junctions, the disturbances in the endothelium permeability contributed to the development of subendothelial layer edema, resulting in its significant thickening. This layer became looser and contained abundant collagen fibrils.

[Organic features of mast cells cytochemistry in rats].

The mast cells from small intestine mesentery, brain membrane, skin, uterus, wide uterical ligament, ovaries and peritoneal fluid were studied in rats by hystochemical and immunocytochemical technique. Constitutional and inducible NO-synthase, Ca2+-ATPase, cytochromoxidase, and biogenic amines (histamine, dopamine, serotonin) were revealed in mast xells, localized in different tissues. A great number of mast cells with positive cytochromoxidase reaction was presented in skin, intestine mesentery and peritoneal fluid, while the amount of must cells containing biogenic amines was found to be greater in the organs with a significant deal of Ca2+-ATPase and NO-synthase-active cells. The number of degranulating mast cells was lower in the organs with a lack of such cells (skin, wide uterical ligament).

[NADPH-diaphorase localization in the radial nerve cords of the starfish Patiria pectinifera]. / Lokalizatsiia NADPH-diaforazy v radial'nykh nervakh morskoi zvezdy Patiria pectinifera.

The presence and localization of NADPH-diaphorase (NADPH-d) in the radial nerve cords of Patiria pectonifera was shown by electron histochemistry. NADPH-d-positive structures were found in ectoneural and hyponeural regions of the radial nerve cord. Ultrastructural localization of NADPH-d was detected in neurons, sensory cells, supporting cells, and in the nerve plexus. The highest enzymatic activity in ectoneural region of the radial nerve cord is due, presumably, to the involvement of NADPH in sensory signal processing.