Tissue-Engineered Constructions in Biophysics, Neurology and Other Fields and Branches of Medicine.

This paper describes the gangliopexy method, a method for creating a new center of local neurohumoral regulation, based on the formation of new connections discovered between the nervous system and the vascular system. The prospects for the development of this method are studied. At the same time, novel concepts about the cycles of nitric oxide and the superoxide anion radical are introduced. A possible role of these cycles is examined in the protection of cells and the body as a whole against oxidative and nitrosative stress, which develops when (in 5-30% of cases) destructive changes in the displaced ganglion lead to vascular complications and an increased risk of mortality. Mechanisms that can protect nerve cells, prevent the development of destructive changes in these cells and reduce the risk of mortality are also investigated.

Causal Relationship between Physiological and Pathological Processes in the Brain and in the Gastrointestinal Tract: The Brain-Intestine Axis.

The brain and gastrointestinal tract are the most important organs responsible for detecting, transmitting, integrating, and responding to signals coming from the internal and external environment. A bidirectional system of neurohumoral communication (the "intestine-brain" axis) combines the activity of the intestine and brain (or brain and intestine) of a person. It affects human development and behavior. This paper analyzes the literature data on the existence of a relationship between the central and enteral nervous systems. Based on data on the number of neurons in the enteral nervous system (approximately 250 million nerve cells), the concept of a "second brain" in the intestine has been proposed in foreign literature, which, by its influence on the brain, can have a more powerful influence than the spinal cord (approximately 10 million neurons) with its autonomic nervous system. However, it turned out that Russian scientists, academicians of the Academy of Sciences of the Soviet Union I.P. Pavlov, K.M. Bykov, and A.M. Ugolev, analyzed cortical-visceral relationships in the 20th century and wrote about the existence of a connection between the central and enteral nervous systems. One of the urgent problems of modern physiology, pathophysiology, biophysics, biochemistry, and medicine is to clarify the causal relationship between the central and enteral nervous systems, as well as between neurological, mental, and gastrointestinal diseases in order to combine the efforts of specialists of various medical and biological profiles to solve urgent medical problems.

[APOΕ gene polymorphism and markers of brain damage in the outcomes of severe traumatic brain injury in children]. / Polimorfizm gena APOΕ i markery povrezhdeniya mozga v iskhodakh tyazheloi cherepno-mozgovoi travmy u detei.

OBJECTIVE: To compare apolipoprotein E (APOE) genotypes with outcomes and levels of neuromarkers in children with severe traumatic brain injury (TBI). MATERIAL AND METHODS: APOE polymorphisms were genotyped in 69 children with severe TBI. The following markers of brain damage were identified: neuron-specific enolase (NSE), glial protein S100b, content of autoantibodies (aAB) to glutamate receptors (to the NR2 subunit of NMDA receptors), aAB to S100b and brain-derived neurotrophic factor (BDNF). RESULTS AND CONCLUSION: There was no association between APOE 3/3, 3/4, 3/2 genotypes and outcomes assessed by the Glasgow Outcome Scale (GOS). The greatest number of favorable outcomes was noted in the group of APOE 3/3 genotype carriers (60%). The ratio of favorable outcomes to unfavorable outcomes was equal (50%:50%) in groups with APOE 3/4 and APOE 3/2 genotypes. An association between APOE polymorphism and BDNF was found: there were normal BDNF levels in the APOE 3/3 group and reduced levels in the APOE 3/2 group. The correlation between neuromarkers and GOS scores was shown for BDNF and aAB to S100b. In children with favorable TBI outcomes, normal BDNF levels and a lower level of aAB to S100b were observed. Regardless of APOE genotypes, almost all children with severe TBI (95%) showed a significant increase in aAB to glutamate receptors in the remote period and most children had an increase in aAB to S100b in the blood. This fact can be explained by the presence of cerebral hypoxia, activation of autoimmune processes and increased BBB permeability, which may be enhanced by increased NO content and intensification of oxidative processes in children with severe TBI.

Molecular Mechanisms of Action of Gas Transmitters NO, CO and H2S in Smooth Muscle Cells and Effect of NO-generating Compounds (Nitrates and Nitrites) on Average Life Expectancy.

Gaseous signaling molecules (gas transmitters) take an especial position among the numerous signaling molecules involved in the regulation of both intracellular processes that occur in different types of cells and cell-cell interactions. At present time, gas transmitters include three molecules whose enzymatic systems of synthesis and degradation, physiological action and intracellular effectors, the change of which under the action of gas transmitters may result in physiological and/or pathophysiological effects are well- determined. These molecules include nitrogen oxide (NO), carbon monoxide (CO) and hydrogen sulfide (H2S). They are involved in the regulation of functions of various organs and systems of the human body, including the circulatory system. Interaction of NO, CO and H2S with various enzymatic and structural components of endothelial and, especially, smooth muscle cells has a significant impact on vascular tone and blood pressure. Furthermore, the crossing of NO-, CO- and H2S-mediated signaling pathways at common effectors and interaction with each other can determine the end, resulting functional response of the cell. The knowledge of the molecular targets of gas transmitters' action, the structure of the binding centers for gas transmitters and their interaction with each other may be essential in the development of methods of regulation of these signaling systems by targeted, directed action. This review summarizes the molecular mechanisms of the NO, CO and H2S interaction with the main targets, which carry out their regulatory effect on vascular smooth muscle cells. Also we describe here different ways of cross-regulation of NO-, CO- and H2S-dependent signaling pathways. We analyzed NO-synthase and nitrite reductase systems of nitric oxide cycle and discuss the nitrate-nitrite background of the existence of modern man, which can substantially modify the signaling system, the metabolism of virtually all cell ultrastructure of neurons, neuron-neuron and neuron-glial interactions and exerts its influence on socially significant diseases that can affect the quality and the average life expectancy.

[Complexes of nitric oxide with hemoglobin and paramagnetic metalloenzymes in the brain and blood of mammals after intermittent hypoxia]. / Kompleksy oksida azota s gemoglobinom i paramagnitnye metallofermenty v mozge i krovi mlekopitayushchikh posle kratkovremennoi gipoksii.

AIM: To investigate an effect of hypoxia on the formation of nitric oxide (NO) in the brain and blood of rats and an influence of L-NNA, the NO-synthase inhibitor, and sodium nitrite on NO generation in separate and simultaneous introduction of these compounds in the animal body. MATERIAL AND METHODS: Experiments were conducted on 42 rats of Krushinsky-Molodkina (K-M) line without adaptation to hypoxia and short-term adaptation to hypobaric hypoxia. Animals were intraperitoneally injected with sodium nitrite (0.5 mg/100 g) and the non-specific inhibitor of NO-synthase L-NNA (2.5 mg/100 g) at different times. Changes in NO content were assessed using electron paramagnetic resonance (EPR) by the intensity of nitrosyl Hb-NO complexes in ERP blood spectra. RESULTS AND CONCLUSION: The production of NO in the brain, compared with that in the blood, was significantly reduced, not only under physiological conditions, but also after the administration of moderate doses of NaNO2. In the short-term hypobaric hypoxia, there was the intensification of the processes of the reduction of NO2- to NO and the formation of nitrosyl heme-NO complexes not only in the blood but also in the mammalian brain.

Gasotransmitters: Physiological Role and Involvement in the Pathogenesis of the Diseases.

In recent decades, it has been found the existence of a new class of biologically active substances - gaseous mediators (gasotransmitters), performing in the cells the signaling function and with high specificity involved in the intercellular and intracellular communication. This review characterizes the main gasotransmitters: nitric oxide, carbon monoxide, hydrogen sulfide, sulfur dioxide and polysulfides. Their physiological role and involvement in the pathogenesis of diseases is described. Basic information about main gasotransmitters is generalized in the original table. Nitrate-Nitrite background, which is a chemical background of the existence of modern man, affects the intra- and intercellular signaling system, alters the ultrastructure of neurons, neuron-neuron and neuron-glia interaction, eliminates the effects of endogenous gasotransmitters and affects the average life expectancy. In accordance with the principle of cyclicity is proposed and substantiated the hypothesis of the existence of "hydrogen sulfide cycle", combining three sulfur-containing gasotransmitters. It is suggested that the cyclic organization of gasotransmitters in the cells and the whole body can be due to the existence of a global principle of cyclicity, which can spread its influence on almost all the structural and functional levels in the animate and inanimate nature.

[The Dual Nature of the Electrical Signals of the Brain (Electrical and Electrochemical) Which Were Recorded With the Help Polarizable Electrodes From Inert Metals].

In the modern neurophysiology opinion was confirmed that the electrical signals of the brain in the frequency band from DC to electroencephalogram recorded with metallic conductors of inert metal implanted in the brain are formed solely by changes in the electric field of the brain. This paper presents a review of the literature and our own data, according to which the formation of these signals involves two factors. One factor is a change in the charge of the electric double layer electrode having a capacitor property and change the value of its charge with changes in the electric field volume conductor--the brain. Another factor is an electrochemical signal is defined by local changes in the redox potential (E) neuronal-glial populations surrounding the electrode. The paper provides an overviews the electrical and electrochemical properties of the electrodes of the inert metals used in electrophysiology. It is shown that each of these factors has the characteristic parameters over time and amplitude. The data of own studies of local changes in E cortex accompanying brain's response to the implantation of electrodes in the brain's cortex, the natural behavior of animals in the wake-sleep, integrative brain function and effect of pharmacological agents. These results give evidence of the highly informative study of local changes in brain E in order to study energy metabolism in the brain of waking animals, and lay the foundation for the study of local changes in brain energy metabolism in free animal behavior.

[Inhibitors of neuronal and inducible NO-syntases enhance the effect of short-term adaptation to hypoxia in rats of Krushinsky-Molodkina strain].

We found that selective inhibitors of neuronal and inducible NOS (7-nitroindazole and aminoguanidine) significantly enhance the protective effect of short-term adaptation to hypoxia on the development of stress lesions in rats Krushinsky-Molodkina.

[Gas Signalling in Mammalian Cells].

At the end of the last century after the discovery of signaling functions of nitric oxide (NO, II), a new class of biologically active substances was admitted. It includes so-called gas transmitters acting as intercellular and intracellular regulators of different physiological functions. Currently, this class includes such gases as NO, carbon monoxide (CO) and hydrogen sulfide (H2S). It was found that these gases regulate not only functions of the. gastrointestinal tract and the cardiovascular system, where it has been determined initially, but also affect the function of the central and peripheral nervous.systems. Apparently, they constitute a single complex of gas transmitters, which easily penetrates through the membrane and regulates numerous enzymatic and non enzymatic cells reactions. This review presents the mechanisms of gas transmitters' influence on the electrical and contractile properties of smooth muscle cells (SMC) as a possible new ways to interact with the "classical" intracellular signaling cascades (Ca2+, cyclic nucleotides) and effectors systems. On account of their interactions the role of cyclic nucleotides and calcium ions in the implementation of the signal gas molecules functions is analyzed. We summarize the literature data and the results of our own research on the role of SMC membrane ion-transporting systems in myogenic effects of NO, CO and H2S and describe possible reasons of gas transmitters multidirectional influence on the excitation-contraction coupling in SMC.

[PLASTIC REORGANIZATION OF THE ULTRASTRUCTURE OF SYNAPSES IN THE CEREBELLUM DUE TO TOXIC EFFECTS OF GLUTAMATE AND NO-GENERATING COMPOUND].

Ultrastructural changes in synapses between parallel fibers (PF) and the spines of Purkinje cell dendrites (PCD) in frog cerebellum were studied after exposure to high concentrations (1 mM) of glutamate (Glu) and NO-generating compound in experimental model. It was shown that exposure to Glu resulted in the envelopment of the terminal bouton by the spine, while under the influence of NO-generating compound, on the contrary, the spine was surrounded by the bouton. Morphological study has shown that in Glu solution there was the predominance of synapses in which the glial cells surrounded the spines, while in the presence of NO they covered the boutons. After the electrical stimulation of PF, the relative number of synapses, containing the boutons surrounded by glial cells, was 10 times higher as compared to those in which the glial cells surrounded the spines. The observed morphological changes reflect the functional state of synapses between PF and PCD in response to the damaging effects of excess Glu and NO, that is expressed in different forms of synaptic contacts and neuronglial structures.

[The influence of inhibitors of neuronal and inducible NO-synthases on experimental hemorrhagic stroke]. / Vliianie ingibitorov neironal'noi i indutsibel'noi NO-sintaz na razvitie gemorragicheskogo insul'ta v éksperimente.

Objectives. To study the effect of inhibitors of neuronal and inducible NO-synthase on the development of hemorrhagic stroke in rats Krushinsky-Molodkina (KM) without adaptation to hypoxia and with short-term adaptation to hypobaric hypoxia. Material and methods. Ninety rats were included in the study. Experiments with short-term adaptation to hypobaric hypoxia were performed on 48 rats. The inhibitor of inducible NO-synthase (aminoguanidine, "Sigma") or the inhibitor of neuronal NO-synthase (7-nitroindasol, "Sigma") were injected in dosage 2.5 mg/100g intraperitoneally. Results. Selective inhibitors of neuronal and inducible NO-synthase had a protective effect on stress injuries in KM rats. The inhibitor of neuronal NO-synthase was more effective than the inhibitor of inducible NO-synthase in the experiments without adaptation to hypoxia. Markedly greater protective effect was achieved by the simultaneous introduction of inhibitors of neuronal and inducible NO-synthase. The greatest protective effect in the development of stress damage in rats of KM was observed in short-term adaptation to hypobaric hypoxia with simultaneous introduction of both inhibitors. Conclusions. It can be assumed that an excessive amount of NO produced by neuronal and inducible NO-synthases during the acoustic exposure in KM rats leads to stress damage. Use of selective inhibitors reduce the excess NO synthesis and the development of audiogenic stress damage caused by hemorrhagic stroke.

Effect of cation type and concentration of nitrates on neurological disorders during experimental cerebral ischemia.

Experiments were performed on the model of ischemic stroke due to bilateral occlusion of the carotid arteries. Nitrates had various effects on the dynamics of neurological disorders and mortality rate of Wistar rats, which depended on the cation type and concentration.

Effects of selective inhibitors of neuronal and inducible NO-synthase on ATP content and survival of cultured rat cerebellar neurons during hyperstimulation of glutamate receptors.

We studied the effects of selective inhibitors of neuronal and inducible NO-synthase (7-nitroindazole and aminoguanidine) and non-selective NO-synthase inhibitor L-NAME on ATP content and survival of cultured rat cerebellar neurons during hyperstimulation of glutamate receptors with toxic doses of glutamate. Application of 100 µM glutamate reduced ATP content in the primary culture of 7-8- and 14-15-day-old cerebellar granule cells by 66 and 49%, respectively, in comparison with the control. Inhibition of nitric oxide synthesis with 7-nitroindazole during glutamate exposure in the culture of 7-8-day-old neurons and with 7-nitroindazole and aminoguanidine in the culture of 14-15-day-old neurons ensured better protection of cells from ATP level decrease than non-specific inhibition with L-NAME. In addition, inhibition of neuronal and inducible NO-synthase during glutamate exposure decreased death of "young" neurons, whereas death of "old" neurons remained high under these conditions.

[The possible role of nitrogen dioxide produced in the field of bifurcation of vessels, in the processes of their damage in hemorrhagic strokes, and the formation of atherosclerotic plaques].

According to modern views the formation of atherosclerotic plaques is associated with accumulation of cholesterol in the vascular wall. This is due to an imbalance between the intake of cholesterol in the intima of vessels, together with the low-density lipoproteins (LDL) and its output with high-density lipoprotein (HDL). Change of LDL (glycosylation, lipid peroxidation, hydrolysis of phospholipids) and the effective release of cholesterol from the endothelium of the vascular wall are the factors that cause an imbalance in cholesterol metabolism. In this paper we propose a new concept of the mechanism of initial formation of atherosclerotic plaques, which can complement the existing concepts. According to this concept an important role in the early stages of atherosclerosis are highly reactive molecules of nitrogen dioxide (NO2), resulting from the violation of the cycles of nitric oxide and superoxide anion radical. Hypothesized that the mechanism of antiradical protection of cells and the organism as a whole, above all, laid out in most of the cyclic organization of metabolic processes that involve the formation of free radicals. Violation of this cyclic mechanism may be one of the causes of many diseases associated with hypoxia/ischemia and inflammation. The review considers the hypothesis of the possibility of participation of NO2 and OH-radicals formed in violation of the cycles of NO and superoxide, in the mechanisms of vascular damage with hemorrhagic stroke and in the formation of atherosclerotic plaques.

Effect of locomotor activity on ultrastructure of cerebellar neurons, neurological disturbances, and survival of Krushinsky-Molodkina rats with hemorrhagic stroke.

We studied the effect of locomotor activity on the ultrastructure of cerebellar neurons, neurological disturbances, and survival rate in Krushinsky-Molodkina rats during the development of hemorrhagic induced by acoustic stress. In animals with high spontaneous locomotor activity, severe edema of cerebellar neurons (resulting in the destruction of surrounding structures) and swelling of the synapses (terminals of mossy fibers on granule cell dendrites) were observed. By contrast, the areas of intracerebral, subdural, and subarachnoid hemorrhages were lower in rats under conditions of forced rest.

[History of physiology physiologists Evgeniy Borisovich Babski (1902-1973)].

The paper analyzes works of eminent physiologists of the twentieth century the Academician of Ukraine SSR, professor Eugene Borisovich Babskii. During 50 years of research in Moscow and Kiev E.B. Babskii published more than 400 works. His main research devoted to investigation of the motility of the digestive tract, general physiology of the nervous system, chemical factors of excitation, mechanisms of muscle contraction, medical electronics and cybernetics and history of human and animal physiology. However, the most significant contribution of Babskii E.B. is his analysis of circulation physiology--the investigation of miocard energy, the physiological effects of electrical stimulation of the heart in the experiment, neural regulation, the ionic mechanisms of automaticity of the heart and myocardial metabolism in different phases of the cardiac cycle. Babskii E.B. and his colleagues firstly created the original method of study of cardiac activity--dinamocardiografy. Academician Babskii E.B. is considered the progenitor of heart electrical stimulation method of Russian Physiology and Medicine. These and many other ideas of Babskii E.B. has been further developed by his students, colleagues and followers.

[The effect of nitrates on the outcome of acute experimental ischemic stroke].

Effects of nitrates NaNO(3), KNO(3), Mg(NO(3)) 2 on animals (Wistar rats) were studied on the basis of the experimental model of ischemic stroke induced by the occlusion of two carotid arteries. The animals were divided into two groups: the main group (n=60) and the control group (n=30). Three series of experiments were conducted. In each experiment, the rats of the main group were treated with one of nitrates and the control group was treated with physiological solution. It has been shown that nitrates exert either positive or negative effect depending on the cation type, nitrate concentration and the duration of their action on the dynamics of neurologic disturbances. Conditions of the development of neuroprotective effect of nitrates are discussed.

[Fusion of frog cerebellar granule cells induced by toxic effects of glutamate and NO-generating compound].

The ultrastructural changes of cerebellar granule cells were studied under stroke modeling conditions, after the toxic effects of glutamate (Glu) and NO-generating compound. Glu toxic doses were shown to induce two types of nuclear chromatin changes. In some cases, the appearance of practically completely decondensed nuclear chromatin was detected, while in the others the nuclei contained partially decondensed chromatin. Pathological fusion of granule cells was observed in both cases. The toxic effect of NO-generating compound on granule cells also caused the appearance of the cells with both completely and partially decondensed (flocculent) chromatin. Granule cells with different chromatin type were able to fuse with each other. Thus, Glu and NO, causing changes in nuclear chromatin, activate the processes of cell clustering with the following cytoplasmic fusion and the formation of multinuclear conglomerates. The possible physiological role of granule cells fusion induced by high concentrations of Glu and NO-generating compound is discussed. This process is considered as a realization of the compensatory-adaptive reactions under extreme conditions observed in the stroke and oxidative stress.

[Autoantibodies to α7-subunit of neuronal acetylcholine receptor in children with traumatic brain injury].

An objective of the study was to search for new biologically significant markers of brain damage. Levels of blood serum autoantibodies (aAB) to different fragments of α7-subunit of acetylcholine receptor (ACR) were studied in children with traumatic brain injury of different severity. The more severe was trauma, the higher was the level of aAB to fragments of α7-subunit of ACR in the first week after trauma. The data obtained suggest that α7-subunits of ACR and aAB to them are involved in the pathogenesis of traumatic brain lesions and, probably, play a significant role in the course of post traumatic period.