The aim of the presented study was to identify the features of the quantitative formation of axoplasmic organelles with a primary analysis of the components of the axonal cytoskeleton (microtubules and neurofilaments) of myelinated and non-myelinated nerve fibers in the dental pulp during the eruption of permanent teeth in humans. Material and methods. The neurovascular bundle of dental pulp was studied in children aged 5, 10 and 14 years. On ultramicrographs of pulp sections at a magnification of ×40000, the diameters of myelin and non-myelin conductors were measured; per 1 µm2 of axoplasm of nerve conductors, the density of axoplasmic organelles, the number of microtubules and neurofilaments were calculated. The reliability of differences between groups was assessed using multivariate analysis of variance (MANOVA), the significance of average values was assessed using the Tukey test, correlation analysis was carried out using the Pearson criterion. Results. The process of eruption of permanent teeth was accompanied by a gradual increase in the number of organelles per unit cross-sectional area of the nerve, regardless of its characteristics (myelinated or nonmyelinated fiber type). The highest density of organelles was observed in large-diameter fibers of both myelinated and non-myelinated conductors throughout the indicated segment of ontogenetic development. The presence of myelin in the nerve sheath correlates positively with microtubule saturation in fibers of large (r=0.267, p=0.001) and small (r=0.314, p=0.000) diameters, and negatively in fibers with medium diameter (r=-0.246, p=0.002). The proportion of neurofilaments represented among the axoplasmic organelles of the neuroplasm as a whole was especially high in myelinated and unmyelinated nerve fibers of small diameter, averaging up to 70%. The presence of the myelin sheath and fiber diameter are not associated with neurofilament density with the exception of medium-diameter fibers (r=0.195, p=0.001). Conclusion. The process of eruption of permanent teeth is accompanied by an increase in the number of organelles, cytoskeletal elements in the axoplasm of nerve fibers innervating the dental pulp, providing the morphological substrate of neuroplasticity, which must be taken into account, for example, in the process of dental implantation and dental prosthetics.

The oxide-ruthenium electrode is used for the analysis of oxygen-containing biologically active organic substances. This electrode can be recommended for the analysis of formic acid in dilute solutions (no more than 2 ml in 100 ml of solution). Due to the presence of two display potentials, the reliability of the determination of formic acid in solution is quite high. For the determination of nitrogen-containing compounds (urea), oxide-ruthenium electrodes are unsuitable. The obvious differences in the polarization curves of magnesium orotate tautomers indicate that the structural differences of tautomers persist in aqueous solutions. An electrochemical method using a ruthenium oxide electrode can be recommended for the analysis of the isomeric state of organic compounds containing OH and COOH groups. The electrochemical method has clearly shown that the difference in the structural state of magnesium orotate tautomers persists in solutions.

The aim of the study was to reveal associations between the number of functioning microvessels and formation of adrenergic control (nerve endings containing catecholamine) of the dental pulp in children and adolescents of Udmurtia during the period of permanent teeth eruption. Material and methods. The study included neurovascular bundle of the pulp of teeth extracted for medical reasons at the age of 5 to 14 years. At the light-optical level, the number of functioning capillaries was counted; an electron microscopic technique was used to qualitative assess the microvascular bed. Identification of catecholamines and counting the number of adrenergic nerve endings were performed histochemically using glyoxylic acid. Results. The process of permanent teeth eruption is accompanied by an intensively increased number of functioning capillaries and an increased density of adrenergic nerve endings. The number of functioning microvessels increases significantly (by 3.36 times) during childhood (from 5 to 9 years); the number of capillaries stabilizes in adolescence. The increased number of nerve endings by the end of the childhood period (9 years) is 45.2 ± 12.4 % compared with the age of 5 years. Notably, adrenergic nerve endings modify qualitatively: a pronounced variety of mediatorcontaining forms is replaced by their isomorphic phenotype. With age, predominant distribution of nerve endings along the vessels of the microvasculature tends to increase. Conclusion. Active formation of the microvascular bed during the eruption of permanent teeth is accompanied by an age-dependent increase in the density of localization and qualitative restructuring of adrenergic nerve endings, which provides full catecholaminergic control from the sympathetic nervous system.

The aim of research is to find out the effect of hyperhomocysteinemia on the regenerative process of liver cells. The histological structure of the liver was studied in Rattus norvegicus Berk white rats (n=25) (control group - 10 animals, comparison group with moderate hyperhomocysteinemia - 15 animals). On histological sections stained with hematoxylin and eosin, the following were calculated: the number of nuclei, the proportion of binuclear cells, the area and the diameter of the nucleus; the nuclear cytoplasmic index was calculated. With the use of immunohistochemical stain by antibodies to detect the expression of the Ki-67 marker (rabbit IgG, 1:200; Cell Marque Corporation, USA), the number and intensity of expression of Ki-67-positive cells in the field of view of the microscope were determined. In the liver of animals with moderate hyperhomocysteinemia, the presence of two processes was revealed at the same time. It is reactive-dystrophic (the presence of periportal leukocyte-lymphocyte infiltrates, the appearance of cells in a state of dystrophy and necrosis) and regenerative (an increase in the core area from 52,51±4,5 to 56,68±5,58 µm, nuclear-cytoplasmic ratio, an increase in the number and intensity of Ki-67+cells expression). The presence of hepatocytes with very large nuclei (polyploid), which make up 12,5 % of the entire population is characteristic of homocysteine-induced liver pathology. Hyperhomocysteinemia, along with a decrease in the number and dystrophy of individual hepatocytes, leads to an increase in the diameter and area of the cell nucleus, an increase in the intensity of proliferation, the appearance of polyploid nuclei, which increases the regenerative potential of the liver and provides a crucial role in the homeostasis of the gland. The findings require further research to determine the "critical point" of transition, which will allow modulation of liver tissue function.

The aim of the study was to analyze the microvasculation of the liver in administration of tautomeric forms of orotic acid for a methionine-induced liver pathology to assess the efficacy of the modified drugs and the potentialof their clinical use.Material and methods. The study included 30 white outbred rats, that were simulated methionine induced hyperhomocysteinemia by the administration of methionine, dosage 0.15 g/100 g of the body we ight, via food during 4 weeks. Animals with hyperhomocysteinemia were divided into 4 groups: animals of group 1 received orotic acid (OA) in the initial form (oxo-tautomer), animals of group 2 received hydroxy–tautomer OA after mechanomodification for 1 hour, animals of group 3 received dihydroxy-tautomer OA after mechanomodification for 6 hours. Histological preparations of the liver were used to evaluate the area of hepatocytes; diameters and areas of the central vein, the interlobular vein and artery, the bile duct, and the Visotto coefficient were calculated.Results. Morphometric data of the hepatic microvascular bloodstream with simulated hyperhomocysteinemia evidenced a general increase in the diameter and area of blood vessels, changes affected the system of blood inflow and outflow at the organ level. Administration of OA had a normalizing effect on the liver bloodflow, but the effects were different: the most pronounced effect was detected in administration of the hydroxy-form of the preparation, this can be explained by a modificationin the dispersion of the preparation (without changing the crystal lattice), an increase in the rate of dissolution in water and aqueous solutions, an increasein the number of functionally active groups in the heterocycle of the hydroxy-form of OA. Thus, the hydroxy-tautomer of orotic acid had the greatest efficacy relating to vasodilation of microvessels of the liver bloodstream in hyperhomocysteinemia; the fact suggesting feasibility of its further study in the clinical environment.

The aim of  the  study  was  to  analyze  the  microvasculation  of  the  liver  in  administration  of  tautomeric forms of orotic acid for a methionine-induced liver pathology to assess the efficacy of the modified drugs and the potentialof their clinical use. Material and  methods. The  study  included  30  white  outbred  rats,  that  were  simulated  methionine induced hyperhomocysteinemia by the administration of methionine, dosage 0.15 g/100 g of the body we ight, via food  during  4  weeks.  Animals  with  hyperhomocysteinemia  were  divided  into  4  groups:  animals  of  group  1  received orotic acid (OA) in the initial form (oxo-tautomer), animals of group 2 received hydroxy–tautomer OA after mechanomodification for 1 hour, animals of group 3 received dihydroxy-tautomer OA after mechanomodification for 6 hours. Histological preparations of the liver were used to evaluate the area of hepatocytes; diameters and areas of the central vein, the interlobular vein and artery, the bile duct, and the Visotto coefficient were calculated. Results. Morphometric  data  of  the  hepatic  microvascular  bloodstream  with  simulated  hyperhomocysteinemia evidenced a general increase in the diameter and area of blood vessels, changes affected the system of blood inflow and outflow at the organ level. Administration of OA had a normalizing effect on the liver bloodflow, but the effects were different: the most pronounced effect was detected in administration of the hydroxy-form of the preparation, this can be explained by a modificationin the dispersion of the preparation (without changing the crystal lattice), an increase in the rate of dissolution in water and aqueous solutions, an increasein the number of functionally active groups in the heterocycle of the hydroxy-form of OA. Thus, the hydroxy-tautomer of orotic acid had the greatest efficacy relating to vasodilation of microvessels of the liver bloodstream in hyperhomocysteinemia; the fact suggesting feasibility of its further study in the clinical environment.

Hyperhomocysteinemia causes stress of the endoplasmic reticulum, which suggests the formation of nucleolar stress. The purpose of this work is to clarify the relationship between structural changes in the nucleus and the region of the nucleolar organizer in hyperhomocysteinemia to prove nucleolar stress in hyperhomo-cysteinemia, which can serve as an additional diagnostic marker of the disease. The object of the study was white mongrel rats with methionine-induced hyperhomocysteinemia. Histologic sections of the liver were stained with hematoxylin and eosin (to assess the histological structure of the organ, hepatocyte nuclei), ammonia silver (to analyze the areas of the nucleolar organizer - AgNORs). Morphometry has allowed to establish that hyperhomocysteinemia decreases the number of nuclei (1,86 times, p

Relevance. The topicality of the work is determined by the wide spread of hypomagnesemia among the people, which makes it necessary to correct it. The aim of the work is to elucidate the cell-mediated response of the thymus mastocytic link to magnesium deficiency and its correction by the mechanoactivated form of magnesium orotate. Materials and Methods . Animals with drug-induced magnesium deficiency (administration of furosemide 30 mg/kg for 14 days) were administered either the initial preparation Magnerot (Magnerot, Vervag Pharma, Germany), or its mechanoactivated form. The level of magnesium in the blood was determined by test systems ARKREY (Japan). The concentration of magnesium in the thymus tissue was determined by the method of emission spectroscopy with inductively coupled (argon) plasma on an atomic emission spectrometer. Density of mastocytes and the indices of degranulation and granulolosis were calculated on paraffin sections of the thymus after coloration with toluidine blue. Results and Discussion . It was shown that furosemide administration the amount of magnesium decreased in the blood (from 1,750,08 to 0,9020,18 mmol/l, p0,05), but increased in the thymus (from 1,60,6 in the control to 3,71,2 mg/l); in the gland tissue, the number of mastocytes of morphotype A decreased and the number of mastocytes of morphotype D, after active degranulation, increased (by 7,1 times, p0,05). The type of mastocyte secretion in hypomagnesemia is represented by the merocrine variant. The administration of the initial magnesium orotate led to an increase in the concentration of magnesium in the blood to 1,150,25 mmol/l, which is 65,7% of the initial level, the amount of magnesium in the thymus remained elevated (3,41,1 mg/l), the number of actively degranulating cells (morphotype D) was increased. Mechanoactivated magnesium orotate restored the concentration of Mg2+ in the blood to 89,1% (1,560,18 mmol/l, p0,05) and decreased in the thymus (to 2,30,7 mg/l), restored the subpopulation of mastocytes saturated with heparin (type A), reduced the number of mastocytes of morphotype D. Conclusion . The mechanoactivated form of magnesium orotate has a normalizing effect on the population of thymic mastocytes, shows pronounced immunomodulatory activity, which allows us to consider it as a potential therapeutic agent for clinical testing in the complex therapy of hypomagnesemia and associated immunodeficiency.

Under the conditions of a chronic methionine diet (daily addition of amino acids to food (0.15 g/100 g) and water (1% solution)) during 2–12 weeks, the dynamics of liver tests, infl ammatory changes in the blood and blood lipids was monitored. It was found that a methionine diet (MD) leads, starting from 4 weeks of MD, to medium hyperhomocysteinemia, an increase in liver enzymes (AsAT – 1.73, AlAT – 1.5 times, p<0.05) and bilirubin (by 62.25%), which indicates the formed hepatopathy. Further (12 weeks of MD), the condition is aggravated by an abnormality of excretory liver function and the development of cholestasis (an increase in alkaline phosphatase by 1.65, bilirubin – by 3.31 times, p<0.05).

The aim is to study macro- and microscopic structure, as well as the cellular composition of the glands of the sphincter zones of large intestine of adults of different age groups.Material and methods. On autopsy material obtained from 30 people, without signs of pathology of the digestive tract of three age groups: 20–29 years, 50–59 years, 90–99 years, the structure of the glandular apparatus of the sphincter zones was studied. The areas of the Gerlach flap, Girsch sphincters, Payr–Strauss, Bally, O'bern–Pirogov–Moutier were considered. Quantitative morphometry was performed on histological preparations stained with methylene blue, followed by fixation in a saturated solution of ammonium molybdenum (picric acid), hematoxylin-eosin, picrofuchsin according to Van Gieson. Methods of parametric statistics based on the Statistica 6.0 program were used for statistical data processing.Results. The analysis of the number, size and cellular composition of the glands of the sphincter zones of the large intestine revealed an increase in both the number of glands and their size in all age groups compared to the proximally adjacent areas of the intestine, on average by 1.3–1.5 times. In a similar range, individual indicators of the number of epithelial cells in the glands of the sphincter zones of the colon increased. At the same time, the cellular composition of the glands of the sphincter zones and adjacent areas of the intestinal wall was similar to neighboring areas, with predominant goblet-shaped epithelial cells (52.9–54.2% of cells on the longitudinal section of the gland) and the presence of absorption cells (29.9–31.2%), undifferentiated – 11.9–13.2% and argyrophilic endocrinocytes – 1.4–5.3%.Conclusion. Against the background of narrowing of the lumen of the large intestine in the area of the sphincters and changes in the nature of the mucous membrane, there is an increase in the size and density of the localization of glands in all age groups. This confirms the thesis about the formation of a protective barrier that provides a local adaptive potential of this area of the intestinal wall, against the background of increased mechanical effects of intestinal masses.

Цель исследования: сравнительная оценка терапевтического эффекта таутомеров магния оротата оксо- и гидрокси-форм в коррекции магниевого дефицита и липидного статуса у магнийдефицитных животных. Материал и методы. У животных формировали лекарственно-обусловленный дефицит магния (введение фуросемида 30 мг/кг 14 дней), затем крысы получали оксо-форму (Magnerot, Верваг Фарма ГмбХ и Ко, Германия) и гидрокси-форму магния оротата (МО), полученную методом механоактивации. В крови анализировали уровни магния, кальция и натрия, рассчитывали скорость компенсации магния, оценивали липидный статус (триглицериды, общий холестерин, липопротеиды низкой, очень низкой и высокой плотности), вычисляли индекс атерогенности. Результаты. При введении гидрокси-формы МО в отличие от оксо-формы, скорость компенсации магниевого дефицита выше (на 14 день составляет 29,2 при оксо- и 77,8 при гидрокси-форме МО), восстанавливается соотношение Ca/Mg. Положительная динамика показателей липидного обмена в сравнении с магнийдефицитными животными отмечается на 14 день введения препаратов магния оротата в обеих группах экспериментальных животных, однако, у крыс с введением оксо-формы уровни общего холестерина (на 13,68, р0,05) и холестерина ЛПНП (в 1,28 раз, р0,05) остаются выше исходных показателей, тогда как в группе с введением гидрокси-формы МО результаты сопоставимы с данными интактного контроля. Различия в уровнях ХС ЛПНП между экспериментальными группами на этом сроке введения препаратов составляют 25,66. Коэффициент атерогенности, рассчитываемый на основе полученных данных, возвращается к исходным показателям в группе, получавшей механомодифицированный магний оротат (гидрокси-форма) и остается повышенным (на 26,14) у животных, получавших исходную оксо-форму препарата. Выводы. Гидрокси-форма магния оротата, полученная методом механоактивации исходного препарата Магнерот проявляет более выраженную биологическую (терапевтическую) активность, что проявляется в опережающем восстановлении уровней магния в крови, скорости компенсации дефицита магния, восстановлении соотношения Mg/Ca, нормализации уровня триглицеридов и холестерина ЛПНП, выравнивании коэффициента атерогенности.

The timeliness of the work is due to the prevalence of magnesium deficiency associated with the use of drugs that contribute to the excretion of magnesium from the body. The aim of the work is to elucidate the cell-mediated reaction of the thymus to magnesium deficiency caused by the administration of furosemide. Magnesium deficiency was modeled by intraperitoneal administration of furosemide to experimental rats. The amount of magnesium and sodium in the blood and thymus tissue was determined by inductively coupled plasma atomic emission spectrometry, the cell composition of the thymus was evaluated on histological sections. It is shown that at furosemide load the amount of magnesium decreases in the blood, but increases in the tissue of the thymus gland. The areas of the structural zones of the thymus (subcapsular zone, cortex, medulla), their percentage; cortical/medullary ratio were calculated. Cell density, lymphocyte count large, medium and small lymphocytes, reticular epithelial cells, macrophages, mast cells, apoptotic cells, thymic corpuscle were counted in each structural zone per unit area (100 μ2). In experimental animals the amount of magnesium in the blood decreases, but in the thymus tissue increases, there is leukocytosis and lymphocytosis, eosinophilia. Revealed histo- and cytostructural morphological rearrangements indicate a change in the functional activity of the gland. It was shown that the furosemide-induce deficiency of magnesium the area of the medulla increases, the number of macrophages and apoptotic elements increases; without affecting on the mast cells, but their secretory activity increases. There are size thymic corpuscle and the number of cells in them increases. Thus, the furosemide load is accompanied by magnesium imbalance, proinflammatory changes induce, is accompanied by a proapoptotic action and stimulates the starting of a macrophage reaction and degranulation of mastocytes in the thymus.

he aim of study was to identify a response of the thymus CD68+ cells population to the introduction of tautomeric forms of magnesium orotate under magnesium deficiency.Material and methods. Magnesium-deficient status in rats was simulated by furosemide loading (30 mg/kg 14 days); then the animals were injected magnesium orotate in the initial oxo-form (Magnerot©, Vervag, Germany) and hydroxy-form obtained by mechanical activation, dosage 50 mg/kg, for 14 days., The density of thymocytes and macrophages was calculated in the subcapsular zone, cortex and brain matter (stained with hematoxylin and eosin) in the thymus, macrophages were studied immunohistochemically using monoclonal CD68 antibodies (Clone PG-M1).Results. The oxo-form of the drug was shown to lead to an increase in the number of CD68+ cells in the cortex of thymus (1.7 compared with the control) and medulla of thymus (2.3 times), an increase in the intensity of immunofluorescence (22.5%), the appearance of the thymus lobule of large CD68+cells in the cortex and the medulla part with a clearly traced intracellular localization of granules. The introduction of mechanically activated hydroxyl-form was accompanied by an increase in the number of CD68+ macrophages in the cortical area (1.5 times), and cortico-medullary partition (in 1.4 times), but did not affect the cerebral area of the thymus. The overall intensity of fluorescent staining reduced to 27.87±2.2 relative units (RU).Conclusion. The tautomeric forms of magnesium orotate have a diverse effect on the macrophage population of the thymus: the introduction of the oxo-form is accompanied by a significant activating effect on the CD68+macrophages in contrast to the hydroxyl form having a moderate modulating effect.

Микроэлементы в медицине 20(4): 51-56 ОРИГИНАЛЬНАЯ СТАТЬЯ БИОЭЛЕМЕНТНЫЙ СОСТАВ ОРГАНОВ ЭКСПЕРИМЕНТАЛЬНЫХ ЖИВОТНЫХ ПРИ ФУРОСЕМИДНОЙ НАГРУЗКЕ Н.Н.Чучкова 1 *, О.М

The purpose of the study is a quantitative analysis of myelin nanotubular apparatus of the conductors included in the neurovascular bundle of the tooth pulp in children born and living in places with high ecological and technogenic load. The object of the study was the neurovascular bundle of the tooth (№ 540), removed for medical reasons, in children aged 5 to 14 years (the period of eruption of permanent teeth), living in conditionally favorable areas of Udmurtia and in places with high ecological and technogenic load. Areas with high ecological and technogenic load were considered to be places near the machine-building plant, poultry farms, large motor transport enterprises and highways, etc. (Izhevsk); location of the largest oil depot in the Republic, asphalt and chemical plants of the Kama region of the Republic (Kambarka); the territory of woodworking, furniture factories, woodworking factory “Red Star” (Mozhga); forestry industry (Uva). The influence of living conditions on the composition and ultrastructural provision of the myelin nerve fibers of the tooth pulp is shown: the number of microtubules (regardless of the fiber diameter) and neurophilaments (for conductors of large and small diameters) in the conditions of ecological and technogenic load is reduced. It is noted that differences in living conditions (places with ecological and technogenic load) do not affect the ultrastructural provision of myelin conductors at the time of eruption of permanent teeth (5 years), but quantitative changes progress over time, starting from the period of second childhood (8-12 years) reaching maximum values by the age of 14. The age of 11 years is a critical period with regard to the definition of the neurotubular apparatus of myelin nerves in children living in places with unfavorable ecological and technogenic load.