[The application of method of flow cytometry in diagnostics of hereditary spherocytosis (eosin-5 maleimid binding test)].

The technique of binding eosin-5 maleimid fluorescent dye with lysine-430 of first extracellular protein bulge of band 3 of erythrocytes' membranes makes it possible to detect the defects of cytoskeleton of erythrocytes as a biological foundation of pathogenesis of hereditary spherocytosis. The samples of peripheral blood from 125 adult persons and 18 children with established absence of hematologic disorders were analyzed The samples of peripheral blood from 19 patients with verified hereditary spherocytosis were analyzed too. The method of flow cytometry was applied to register the average intensity of fluorescence of eosin-5 maleimid. The decrease of average intensity of fluorescence of eosin-5 maleimid of erythrocytes of patients with hereditary spherocytosis as compared with data from comparison groups was established in all cases.

[Systemic control of the molecular, cell, and epigenetic mechanisms of long-lasting consequences of stress].

Based on M.E. Lobashev's views of the systemic control of genetic and cytogeneitc processes and a substantial effect of excitability on plastic changes in the central nervous system (CNS), the effect of prolonged emotional and pain stress (PEPS) on the molecular, cell, and epigenetic mechanisms of injury memory was studied in rat strains bred for a certain excitability of the nervous system. PEPS was for the first time found to cause long-lasting (2 months) morphological alterations of the CA3 region of the hippocampus and to modify the genome activity of its pyramidal neurons. The two phenomena were potentiated by a genetically determined low functional state of the CNS. The post-stress regulation of the genome function in hippocampal neurons was mediated by changes in heterochromatin conformation, activation of methyl-CpG-binding protein (MeCP2) synthesis, and subsequent changes in acetylation of histone H4. Genetically determined high excitability of the nervous system proved to be a risk factor that affects the specifics and time course of the observed molecular, cell, and genetic transformations of neurons. The results provide for a better understanding of the epigenetic mechanisms of injury memory, which forms a pathogenetic basis for posttraumatic stress disorder and other human psychogenic conditions characterized by a prolonged duration.

Effect of prolonged emotional and pain stress on the content of methylcytosine-binding protein MeCP2 in nuclei of hippocampal neurons in rats with different excitability of the nervous system.

In rats with low excitability threshold of the nervous system demonstrating significant and persistent behavioral disorders under stress conditions, the content of methylcytosine-binding protein MeCP2 in neuronal nuclei of hippocampal field CA3 decreased over 2 weeks after long-term emotional and pain stress. It was hypothesized that protein MeCP2 triggers epigenetic changes in DNA that underlie "stress memory".

Effect of long-term mental and pain stress on the dynamics of H4 histone acetylation in hippocampal neurons of rats with different levels of nervous system excitability.

Priority data on specific effect of long-term mental and pain stress on the dynamics of H4 histone acetylation in the pyramidal neuron nuclei of the hippocampal CA3 field in rats selected by the nervous system excitability were obtained using a comparative genetic method. The congruency of long-term post-stress modification of H4 histone acetylation in neurons of rats with high threshold excitability and behavioral changes intrinsic of these rats suggest that increased acetylation of H4 histone together with changes in heterochromatin conformation play a triggering role in long-term modifications of genome expression underlying the pathogenesis of posttraumatic stress disorders and other psychogenias.

[Characteristics of heterochromatin of heterophase nuclei from interphase neurons of various brain structures selected for the nervous system excitability]. / Kharakteristika geterokhromatina interfaznykh iader neironov razlichnykh struktur golovnogo mozga krys, selektirovannykh po vozbudimosti nervnoi sistemy.

Quantitative characteristics (the area and number of chromocenters) of the interphase C-heterochromatin in the nuclei of pyramidal neurons of the midbrain reticular formation, sensorimotor cortex, and hippocampus (CA3) of rat strains with different genetically determined excitability were studied in the normal state of the animals and after exposure to a short-term emotional pain stress. The results indicate a relationship between the excitability of the nervous system and structural-functional state of the neuronal interphase heterochromatin. The role of cytogenetic features of different brain structures in the CNS functioning and behavior and their relation with genetically determined excitability of the nervous system are discussed.

45Ca exchange in rat cortex slices in conditions of long-term potentiation.

Existing data on the role of Ca2+ ions in the development of long-term potentiation were used as a basis for studying changes in different Ca2+ compartments in cells in living rat olfactory cortex slices during potentiation. The kinetics of 45Ca2+ exchange were studied at 5, 15, and 30 min of potentiation. During the induction phase (1-5 min) of long-term potentiation, the fraction of tightly-bound intracellular Ca2+ decreased. There were no changes in the content of Ca2+ ions in other fractions at this stage. During maintenance of potentiation, which lasted 15-25 min, Ca2+ levels in the extracellular and intracellular compartments did not differ from controls. At 30 min, during extinction of long-term potentiation, there was a significant redistribution of Ca2+ in cells: the levels of free and loosely-bound Ca increased, as did extracellular Ca2+.

[45Ca metabolism in slices of rat cerebral cortex during long-term potentiation]. / Obmen 45Ca v srezakh kory mozga krys v usloviiakh dolgovremennoi potentsiatsii.

Ca45 kinetics was studied in 5, 15 and 30 min after potentiation. In the induction phase (1-5 min), the potentiation decreased the fraction of intracellular bound Ca. The 15-25 min potentiation the intra- and extracellular Ca levels was equal to the control ones. In 30 min, a considerable redistribution of Ca in the cells occurred.

Number of correlates of membrane metabolism and long-term potentiation in rat brain slices.

Long-term potentiation was elicited in living slices of rat olfactory cortex by stimulation of the lateral olfactory tract. A group of interdependent parameters of membrane metabolism was studied, i.e., the kinetics of 45Ca metabolism, lipid peroxidation, and antioxidant defense; cytochemical measurements were made of Na+, K(+)-ATPase activity in neurons and glial cells; the functional (GTPase) activity of G-proteins was also studied. All parameters were compared with the bioelectrical activity of slices at three time points after tetanization, i.e., 3-5, 15, and 30 min. In most cases, regular phasic changes in metabolic parameters occurred, and their functional significance is discussed.

[The effect of mexidol on the dynamics of the normalization of the neurochemical and behavioral parameters of rats after deprivation of the paradoxical sleep phase]. / Vliianie meksidola na dinamiku normalizatsii neirokhimicheskikh i povedencheskikh parametrov u krys posle lisheniia paradoksal'noi fazy sna.

Using cytophotometric method it was shown that intraperitoneal injection of mexidol in the dose of 50 mg/kg resulted in a significant accumulation of proteins and RNA in cells of dorsal raphe nucleus (DRN) and locus coeruleus (LC) 6 hs following the injection. In rats deprived of paradoxical sleep for 24 hours, the effect of mexidol prevented the decrease in DRN proteins and RNA and even increased them, and prevented their decrease in LC. In the postdeprivation period, the beneficial effect of mexidol manifested itself in the acceleration of normalizing the levels of proteins and RNA in DRN and LC. The antistress effect of mexidol manifested itself also in the acceleration of normalizing the rat memory in the restoration period following the deprivation of paradoxical sleep. The normalization of metabolism and functional activity in serotonergic DRN is suggested to determine to a considerable extent the functional state of cerebral structures participating in the control of memory as well.

[The dynamics of the normalization of the behavioral and neurochemical disorders evoked in rats by deprivation of the paradoxical sleep phase]. / Dinamika normalizatsii nekotorykh povedencheskikh i neirokhimicheskikh narushenii, vyzvannykh u krys deprivatsiei paradoksal'noi fazy sna.

The 24-hr deprivation of paradoxical sleep interferes with the passive avoidance response in rats and decreases the absolute amount of proteins in neurons and gliocytes of the raphe nucleus' dorsal portion while increasing the amount of proteins and nucleic acids in the locus coeruleus' cells. The protein metabolism in serotoninergic cells of the raphe nucleus seems to be involved in the processes of memory and behaviour.