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.

Chitosan-g-oligo(L,L-lactide) copolymer hydrogel for nervous tissue regeneration in glutamate excitotoxicity: in vitro feasibility evaluation.

Over the last decade, a number of hydrogels attracted great attention in the area of brain tissue engineering. The hydrogels are composed of hydrophilic polymers forming 3D network in water. Their function is promoting structural and functional restoration of damaged brain tissues by providing mechanical support and navigating cell fate. This paper reports on the neurocompatibility of chitosan-g-oligo(L,L-lactide) copolymer hydrogel with primary rat cortical neuron culture. The hydrogel was produced by a molding technique on the base of photocurable composition consisting of chitosan-g-oligo(L,L-lactide) copolymer, poly(ethylene glycol) diacrylate and photosensitizer Irgacure 2959. The influence of the hydrogel on cell viability, phenotype and calcium homeostasis, mitochondrial potential and oxygen consumption rate in glutamate excitotoxicity was analyzed using primary neuron cultures obtained from a neonatal rat cortex. This study revealed that the hydrogel is non-cytotoxic. Dissociated neonatal rat cortical cells were actively attaching to the hydrogel surface and exhibited the phenotype, calcium homeostasis and mitochondrial function in both standard conditions and glutamate excitotoxicity (100 µM) similar to the control cells cultured without the hydrogel. To conclude, in this study we assessed the feasibility of the application of chitosan-g-oligo(L,L-lactide) copolymer hydrogel for tissue engineering therapy of brain injury in an in vitro model. The results support that the hydrogel is able to sustain realization of the functional metabolic activity of neonatal rat cortical cells in response to glutamate excitotoxicity.

[Biomarkers in children with traumatic brain injury].

OBJECTIVE: to study the content of biomarkers of diagnostic and prognostic value in the assessment of traumatic brain injury (TBI) severity in children. MATERIAL AND METHODS: Authors determined the levels of glial protein S100B, neuronspecific enolase (NSE), autoantibodies (aAb) to glutamate receptors and natural autoantibodies (nAb) to S100B and brain-derived neurotrophic factor (BDNF) in serum/plasma of children with different outcomes of TBI. All parameters were analyzed in the 1-3rd, 7-8th, 14-15th and 20-23rd days after TBI, and, in some cases of severe brain injury and long stay patients in hospital, in 11-12 months after TBI. The severity and outcome of TBI were assessed according the Glasgow coma scale (GCS) and the Glasgow outcome scale (GOS), respectively. RESULTS AND CONCLUSION: The content of NSE and S100B increased immediately after TBI regardless of TBI severity, but in cases with favorable outcome it dropped to a normal level in the first 3 days. The maximum levels of S100B protein and NSE were observed in children with fatal TBI, and higher values were observed throughout the post-traumatic period. The low levels of aAb to NR2-subtype of glutamate receptors that were similar to controls and the high level of nAb to S100B in the first days after severe TBI indicate the failure of compensatory-adaptive immunological mechanisms and the high permeability of the brain-blood barrier, which were poor prognostic signs for children with severe TBI. Mild and severe TBI with full recovery were accompanied by higher values of ВDNF in the 1st day followed by a decline to the 3rd day. The level of BDNF in the 1st day of TBI was the lowest and subsequently continued to decline in patients with severe TBI with fatal outcome.

[Peripheral Blood Lymphocytes Mitochondrial Function in Children with Traumatic Brain Injury].

BACKGRAUND: It is known that mitochondria play an important role in the mechanisms of brain cells damage and death following traumatic brain injury (TBI). However, the relationship between the severity of brain damage following TBI and mitochondrial dysfunction are not well defined. AIM: to study activities of NADN- and succinate dehydrogenases, a key enzyme of mitochondrial oxidative phosphorylation in children with TBI of varying severity and different outcomes; to detect ATP content in lymphocytes; the level of NOx and 3-nitrotyrosine in serum and plasma. Methods: all parameters were determined in the dynamics of one month following TBI, and in some cases up to the death ofpatients. The severity of TBI was scored by Glasgow Coma Scale (GCS), the outcome of TBI-Glasgow Outcome Scale (GOS). Based on the clinical examination children with TBI were divided into 3 groups: (1) mild TBI; (2) severe TBI and (3) severe TBI with fatal outcome. RESULTS: we found that activity of dehydrogenases is significantly reduced only in patients with the poor neurologic outcome. The greatest decrease in these parameters was observed in patients with severe traumatic brain injury and fatal outcome. A direct correlation was found between the indices of dehydrogenases activity and A TP content in lymphocytes (r = 0.97, p = 0.005). The levels of NOx metabolites and 3-nitrotyrosine were significantly increased in children with severe TBI. CONCLUSION: obtained results suggest that mitochondrial dysfunction, impaired cerebral energy metabolism and oxidative stress contribute to cell death in the brain and thus represent therapeutic targets for the treatment of TBI.

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.

[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.

[S100B protein and autoantibodies to S100B protein in diagnostics of brain damage in craniocerebral trauma in children].

Levels of antibodies AB (AB) to S100B and S100B protein were studied in the blood serum of children with different severity and outcomes of traumatic brain injury (TBI) from the 1st to 15-75th days after TBI. Severity and outcomes were assessed using the Glasgow Coma Scale (GCS). Patients were stratified by outcomes into the following groups: complete recovery (group 1), moderate disability (group 2), high disability (group 3), vegetative state (group 4) and fatal outcome (group 5). In patients of groups 1-3, the changes of S100B in the blood serum didn't depend on the severity of brain's damage; the significant increase of S100B protein levels in the 1st day was accompanied by the decrease to the normal range in the following 2-3 days. On the contrary, the levels of nAB in these groups increased starting from 3-5 days corresponding to the severity of brain's damage. The development of vegetative state was accompanied by low S100B and high AB to S100B levels in the blood serum. The maximal level of S100B protein and increased levels of AB were observed in patients with fatal outcome. In most patients with combined TBI, the levels of both parameters were higher compared to those with separate TBI.

[EPR study of the influence of hypoxia on nitric oxide formation in the blood of Krushinskii-Molodkina rats].

The influence of hypoxia on nitric oxide formation in the blood of Krushinskii-Molodkina rats has been studied by electron paramagnetic resonance. It was found that nitric oxide synthesis in Krushinskii-Molodkina rats is increased compared with that in Wistar rats. A significant enhancement of the EPR signal of Hb-NO complexes in the animal blood was observed after hypoxia simulating the altitude of 5000 m above the sea level, in particular in the presence of sodium nitrite and the NO-synthase inhibitor L-nitroarginine. It was assumed that NO synthesis and nitro-/nitrite- reductase systems are activated under hypoxic conditions.

Autoantibodies to glutamate receptors and products of nitric oxide metabolism in serum in children in the acute phase of craniocerebral trauma.

Levels of serum autoantibodies (aAb) to glutamate receptors and products of nitric oxide (NO) metabolism, i.e., nitrates and nitrites, were assayed in children with recent craniocerebral trauma (CCT) of different levels of severity. All the children showed increases in serum aAb to both AMPA and NMDA receptor subtypes from day 1 to day 10 after trauma. The highest levels of serum aAb were to the NMDA subtype of glutamate receptor, which was characteristic of children with mild CCT (MCCT), with Glasgow Coma Scale (GCS) scores of 14-15 points. Levels of aAb to NMDA (NR2A) receptors in children with severe CCT (SCCT, GCS < 9 points) were lower than in children with MCCT, the lowest levels being seen in the group of children with lethal CCT (SCCT-2). Serum concentrations of NO metabolites increased by large factors in the group of children with SCCT, indicating marked brain hypoxia.

[Autoantibodies to glutamate receptors and metabolic products of nitric oxide in blood serum of children in the acute period of brain trauma].

Autoantibodies (AB) to glutamate receptors of AMPA (Glur1) and NMDA (NR2A) types and nitric oxide metabolites, nitrates and nitrites (NOx), were studied in the blood serum of children with brain trauma of different severity. The level of both AB types increased from the 1st to the 10th day after trauma. The level of NMDA (NR2A) AB was higher comparing to AMPA (Glur1). Patients with mild brain trauma, scoring 14-15 on the Coma Glasgow Scale, had the highest AB concentration while patients with severe brain trauma (scores <9), had the lower level of NMDA (NR2A) AB. The lowest level of AB and the highest level of NOx in the blood serum were found in a group of children with the fatal outcome of severe brain trauma. The many-fold increase of NOx concentration in this group points to marked hypoxia after severe brain trauma.

[Autoimmune mechanisms of modulation of the activity of glutamate receptors in children with epilepsy and craniocerebral injury].

The role of glutamate receptors and their hyperstimulation in the development of autoimmune processes is discussed with reference to brain pathology associated with hypoxia and ischemia. Epilepsy, paroxismal condition, and craniocerebral injury (CCI) in children are shown to be accompanied by a rise in the levels of antibodies against AMPA and NMDA receptors of glutamate and nitric oxide markers (cGMP, nitrates + nitrites). Also enhanced in epilepsy and paroxismal condition are the levels of cGMP and antibodies against AMPA(GluR1) receptors of glutamate. Acute CCI period is characterized by a marked change in the levels of NO metabolites and antibodies to two subtypes of glutamate receptor, AMPA and NMDA. The levels of antibodies to NMDA(NR2A) receptors are significantly different within 1 day after CCI depending on its outcome. Unfavourable outcome of CCI is associated with the lowest level of antibodies and high NO metabolite content. Relationship between the levels of NO and antibodies against glutamate receptors is discussed with the use of experimental data. It is concluded that antibodies to glutamate receptors and receptor hyperstimulation play an important role in pathogenesis of hypoxia.

[Nitric oxide (NO) and NO cycle in myocardium: molecular, biochemical and physiological aspects].

The article continues the series of our publications on the problem of nitric oxide (NO) and its cyclic conversion in mammals. This review is held to analysis of nitric oxide role in regulation of cardiovascular system and in alocation of NO-synthases in myocardium. Molecular, biochemical and cytophysiological aspects that linked, with spatial localization of NO-synthases and mechanisms of NO content regulation in myocardium are considered. The results of author's investigations along the cyclic convertion of NO and literature data about compartmentalization of NO-synthases in myocardium are included in this paper. The contradictory and dissimilar facts about regulatory and toxic role of nitric oxide in cardiovascular system are represented.

[The clinical economic analysis of the methods of ischemic heart disease diagnostics].

The clinical economical analysis was applied to assess the application of different techniques of ischemic heart disease diagnostics - the electro-cardiographic monitoring, the treadmill-testing, the stress-echo cardiographic with dobutamine, the single-photon computerized axial tomography with load, the multi-spiral computerized axial tomography with coronary arteries staining in patients with different initial probability of disease occurrence. In all groups, the best value of "cost-effectiveness" had the treadmill-test. The patients with low risk needed 17.4 rubles to precise the probability of ischemic heart disease occurrence at 1%. In the group with medium and high risk this indicator was 9.4 and 24.7 rubles correspondingly. It is concluded that to precise the probability of ischemic heart disease occurrence after tredmil-test in the patients with high probability it is appropriate to use the single-photon computerized axial tomography with load and in the case of patients with low probability the multi-spiral computerized axial tomography with coronary arteries staining.

Glutamate receptor autoantibody concentrations in children with chronic post-traumatic headache.

We report here studies on the levels of autoantibodies (aAb) to AMPA glutamate receptors (GluR1 subunit) and NMDA glutamate receptors (NR2A subunit) in serum from 60 children aged 7-16 years with chronic posttraumatic headache (CPTHA) following mild craniocerebral trauma (CCT). The first group consisted of 48 children who had sustained cerebral concussion (CC), of which 34 had single-episode CC (subgroup 1a) and 14 had repeated CC (subgroup 1). The second group included 12 children with mild cerebral contusions (MCC). Serum glutamate receptor aAb levels were measured six months and one year after trauma. Increased aAb levels were expressed as percentages and were regarded as significant when increases were to 120% of the level seen in healthy children of the same age. The highest levels of aAb to NMDA receptors were seen in children with MCC (165 +/- 34%) and single CC (145 +/- 12.6%). Children with repeated CC had NMDA receptor aAb at normal levels (108 +/- 12.4%). Increases in NMDA receptor aAb were seen during the first year after trauma. Increases in AMPA receptor aAb were seen in children with repeated CC and MCC (150 +/- 16.8% and 167 +/- 31.3%). EEG studies showed that 18% of these children had nonspecific paroxysmal changes and 6% showed epileptiform activity. These results provide evidence that children with post-traumatic headache demonstrated hyperstimulation of glutamate receptors and overdevelopment of the autoimmune process. Increases in serum levels of aAb to NMDA glutamate receptors reflected hypoxic-ischemic brain lesions in children with CPTHA and dictate the need for these children to receive metabolic therapy.