[Biologically Active Peptides Isolated from Dill Anethum graveolens L].

Peptide mixtures with molecular weights of 1000-2000 Da and in vivo membrano-trophic activity against mouse hepatocyte culture at very low concentrations were isolated from dill Anethum graveolens L. leaves. It has been found that plant peptides in aqueous solution formed larger nanosized particles of approximately 90 nm with a secondary structure mainly composed of ß-structures and random coil structures. We demonstrated that peptides isolated from A. graveolens in vitro at an ultra-low dosage affected the size of the area of pigmented cells of amphibian liver, which are analogous to Kupffer cells of the mammalian liver, using roller organotypic newt liver culture models.

[Identification of a new bioregulator acting in ultralow doses in bulb onion (Allium cepa L.)].

A bioregulator that has physicochemical and biological properties similar to a group of bioregulators isolated from various animal tissues has been found in the bulb onion (Allium cepa L.). It was determined that the biological action of the plant bioregulator is determined by a peptide with molecular weight of 4036 +/- 2 Da whose 18-C-terminal amino acid sequence consisted of 18 residues. On models of seed germination of some vegetable cultures, the ability of the bioregulator isolated from supernatant of onion extract in ultralow doses (10(-13) mg of protein/ml) to inhibit growth and development was demonstrated.

[Study of a new group of bioregulators isolated from the greater plantain (Plantago major L.)].

Proteins with physicochemical properties and biological activity similar to those of membrano-tropic homeostatic tissue-specific bioregulators that had been found earlier in various animal tissues were discovered in leaves of the common plantain (Plantago major L.). To study the specific activity of these plant proteins, we developed an experimental model for organotypic roller cultivation of newt (Pleurodeles waltl) skin tissue in vitro. We showed that the plant proteins of interest exert the wound-healing effect, which is characteristic of this plant, on the skin of vertebrates both in vitro and in vivo.

[MALDI-TOF mass spectrometric identification of novel intercellular space peptides].

We performed the matrix-assisted laser desorption/ionisation, time-of-flight mass spectrometry (MALDI-TOF) analysis of the peptides entering into the composition of not yet explored bioregulators derived from the extracellular matrix of the tissues of the various organs of the mammals, and also plants and fungi. The study included 15 different mammalian tissues, 13 species of plants, and 2 species of fungi. Exploring the bioregulators derived from eye tissues, we demonstrated that their composition includes peptide components with the same values of the molecular weight. The composition of the bioregulators derived from the tissues of various organs of mammals or different species of plants and fungi includes the peptides with different values of molecular weight. Obtained data indicate the growing evidence of the assumptions about the major function of the bioregulators of this group--their involvement in the regulation of tissue-organ homeostasis in the biological systems.

Participation of RNA synthesis in the process of the development and maintenance of conditioned reflexes (an investigation using antitheines).

It has been established that an increase in RNA synthesis in the neurons of the cerebral cortex of rats at the stage of consolidation is manifested in well-trained animals more strongly than in poorly trained animals. The selective influence of propylnorantitheine and demethylated derivatives of ethylnorantitheine on the maintenance of conditioned reflexes has been demonstrated. The effects of these substances on consolidation and long-term memory correlate with the change in the RNA-synthesizing activity of neurons during the effect both in systemic experiments and with the direct interaction with the chromatin of the neurons. The participation of the RNA synthesis of cerebral cortical neurons in the mechanisms of long-term memory is discussed.

Involvement of the genetic apparatus in memory formation mechanisms: the role of the neuronal calcium-regulatory system in rats.

Stimulators of long-term memory (ethylnorantiphein and its analogs M1 and M2) were used to study the dynamics of several components of the neuronal calcium-regulatory system in the rat cortex and hippocampus. There were no changes in the activity of the Mg, Ca-ATPase transporter and actomyosin-like Ca-ATPase in synaptosomes 5, 15, 60, and 180 min after dosage with these agents. On exposure to ethylnorantiphein, M1, and M2, activation of RNA transcription at 60 min was accompanied by notable increases in chromatin Ca-ATPase activity, along with an increase in the synthesis of synaptosomal proteins at 180 min, with an increase in synaptic membrane protein kinase C activity. An increase in chromatin Ca-ATPase activity was also seen during fixation of a conditioned active escape reflex. It is suggested that the increase in protein kinase C activity is associated with secondary rearrangements of the synaptic membranes. The question of the role of direct activation of the genetic apparatus by neuroactive substances in the molecular mechanisms of memory formation is discussed.

Status of the "protein kinase CK2-HMG14" system in age-related amnesia in rats.

The experiments described here demonstrate that disruption of the phosphorylation of transcription factors of the HMG cAMP/Ca-independent protein kinase CK2 class may be the cause of decreased gene expression in age-related cognitive deficits. Amnesia for a conditioned passive avoidance reaction (CPAR) in aged rats (24 months old) was accompanied by decreases in the synthesis of synaptosomal proteins and transcription in nuclei isolated from cortical, hippocampal, and striatal neurons. There was a decrease in chromatin protein kinase CK2 activity and a significant decrease in the phosphorylation of HMG14 by protein kinase CK2. Selective activators of protein kinase CK2 (1-ethyl-4-carbamoyl-5-methylcarbamoylimidazole and 1-ethyl-4,5-dicarbamoylimidazole) increased HMG14 phosphorylation by protein kinase CK2, increased transcription, increased the synthesis of synaptosomal proteins, and decreased amnesia for the CPAR in aged rats. Thus, activation of the "protein kinase CK2-HMG14" system is accompanied by optimization of synaptic plasticity in aged animals. The results provide evidence for the high therapeutic potential of protein kinase CK2 activators.

[Therapeutic potential of protein kinase CK2 modulators]. / Terapevticheskii potentsial moduliatorov proteinkinazy CK2.

Data on the nuclear cascade of signal transduction, including protein kinase CK2 (PKCK2), transcription factor HMG14 and chromatin myosin-like proteins, are generalized with regard for the modern understanding of mechanisms of synaptic plasticity. The role of the neurospecific isoform and subunit structure of PKCK2, of the individual subunit autophosphorylation of PKCK2, of phosphorylation of substrate-proteins in the enzyme activity and of conformation transformations of chromatin is examined. Data on changes in the CK2-induced cascade and synaptic plasticity in learning, on age-related amnesia and on cognitive deficits induced by ethanol and chloridine in rat embryos are presented. The prospects for using modulators PKCK2, 4,5-di(N-methylcarbamoyl)-l-alkyl-imidazoles, as potential nootropics are discussed.

[Current aspects of the search of effective nootropic drugs]. / Sovremennye aspekty poiska éffektivnykh nootropnykh sredstv.

The authors review different mechanisms of mnemotropic and cerebroprotective effects of nootropic drugs. The data concerning the molecular mechanisms of action of the structural analogs of the memory stimulant ethylnorantifeine (etimizol) have been summarized and analyzed. It is shown that the effects of antifeines on the retention of the conditioned reflexes are independent of their effects on the cAMP system and structural-functional condition of the neuronal membrane. The key role in the action of these compounds on the long-term memory is played by the activity of the genetic apparatus. The existence of nootropic receptors in neuronal chromatin is assumed.

[The participation of the genetic apparatus in the mechanisms of memory trace formation: the role of the calcium-regulatory system of the neurons in rats]. / Uchastie geneticheskogo apparata v mekhanizmakh sledoobrazovaniia: rol' kal'tsii-reguliatornoi sistemy neironov krys.

The dynamics of changes in some components of the calcium-regulated system of cortical and hippocampal neurons under the influence of long-term memory enhancers (ethylnorantifein and its analogues M1 and M2) was studied in rat brain. No change was found in the activity of transport Mg, Ca-ATPase and actomyosin-like Ca-ATPase in synaptic membranes 5, 15, 60, and 180 min after the injection of memory enhancers. The activation of the RNA transcription (60 min after the injection) was accompanied by an appreciable increase in activity of the chromatin Ca-ATPase. The amplification of synaptosomal protein synthesis (180 min) was accompanied by an increase in the activation of protein kinase C of synaptic membranes. The increase in Ca-ATPase activity of chromatin was also shown during the consolidation of the conditioned active avoidance in rats. The increase in the activity of protein kinase C seems to be connected with secondary rearrangement in synaptic membranes. The role in the long-term memory is discussed of direct activation of the genetic apparatus by neuroactive substances.

[The participation of RNA synthesis in the process of the acquisition and retention of conditioned reflexes (a study using antifeins)]. / Uchastie sinteza RNK v protsesse vyrabotki i sokhraneniia uslovnykh refleksov (issledovanie s ispol'zovaniem antifeinov).

RNA-synthesizing activity of neuronal nuclei in the neocortex of rats increases after the termination of conditioning depending on the degree of learning. RNA synthesis shifts induced by propylnorantifein and the demethylated derivatives of ethylnorantifein are correlated only with the influence of the drugs on the retention but not the learning. Participation of RNA synthesis by the neurons of the neocortex in the mechanisms of long-term memory is discussed.

[The effect of structural analogs of ethylnorantifeine on the autophosphorylation of cAMP-independent protein kinases of neuronal chromatin]. / Vliianie strukturnykh analogov étilnorantifeina na autofosforilirovanie tsAMF-nezavisimykh proteinkinaz khromatina neironov.

A significant increase in autophosphorylation of cAMP-independent protein kinase No. 11 of brain neuronal chromatin occurs in the presence of ethylnorantifeine and its demethylated analog M1 rather than allyl- and propylnorantifeine. The increase is accompanied by phosphorylation of beta-regulatory and alpha-catalytic subunits of protein kinase No. 11. The nature of the direct action of antifeines on this enzyme correlates with their effects on gene activity and long-term memory storage. Whether neuronal chromatin protein kinase No. 11 is the action target of antifeines in displaying their mnemic effects is discussed in the paper.

[Skeletal muscle nuclear ATPase under denervation]. / ATP-azy iader skeletnykh myshts pri denervatsii.

Denervation of the rabbit gastrocnemius muscle is shown to result after 18 and 72 h in a decrease of Mg2+-, Ca2+-dependent and an increase of Mg+-dependent parts of nuclear ATPase activity, with the total level of the enzyme activity retained. 7 days later the total ATPase activity of nuclei decreases as well as the expense of its EGTA-dependent part. These changes correlate with those in the nuclear Ca total content. Indirect electrostimulation of the denervated muscle increase both the 7g2+-, Ca2+-dependent ATPase activity and the Ca content in nuclei. Tenotomy for 18 h does not change these parameters.

[Comparison of superslow activity with changes in the function of the neocortical cytostructures]. / Sopostavlenie sverkhmedlennoi aktivnosti s izmeneniiami funktsional'nogo sostoianiia tsitostruktur neokorteksa.

Effects of ethimizole on the infraslow ascillations of the brain electrical activity were correlated with the pattern of its distribution within cellular structures. A negative wave of the infraslow activity was found to occur after the drug administration at the moment of its maximal inclusion into the two cellular fractions: the nuclei and the surface proteins of the endoplasmatic net. These structures seem to play part in changes of the central nervous system's functional state.

[Effect of taurine on 45Ca2+ binding by the microsomal membranes of brain cells]. / Vliianie taurina na sviazyvanie 45Ca2+ mikrosomal'nymi membranami kletok golovnogo mozga.

Effect of taurine o binding of 45Ca2+ and 8-aniline-I-naphthaline culfonate by microsomes of rat brain cells was studied. Increased accumulation of Ca2+ in phospholipids of microsomes as well as a distinct decrease in the number of sites for binding of 8-aniline-I-naphthaline sulfonate were observed in presence of taurine. Activity of Ca2+-ATPase was simultaneously increased. The data obtained suggest that the effect of taurine on biomembranes is related to local alterations in their surface charge.

[Participation of adenosine receptors in regulating the functional activity of neuronal chromatin by antifeines]. / K voprosu ob uchastii retseptorov adenozina v reguliatsii antifeinami funktsional'noi aktivnosti khromatina neironov.

The regulatory pathways of chromatin transcription in neurons have been studied. The metabolic events (adenylate cyclase, Ca/phospholipid-dependent protein kinase C) involved in signal transduction via adenosine receptors were investigated with the application of pharmacological tools--the memory stimulant ethylnorantifeine and its structural analogs. It has been modelled the biochemical condition of A1 and A2 adenosine receptors in the cortex and the striatum of rat brain. The antifeines did not influence AC of A1 or A2 receptors, unlike the non-metabolizing adenosine derivatives -PIA, NECA and isobutylmethylcanthine. No direct effect of antifeines (10(-7)-10(-3) M) on membrane-bound protein kinase C was established. The data obtained failed to provide evidence for the antifeine effect on the functional activity of chromatin with regard to the triggering role of membrane acceptor systems. The possibility of direct control of the chromatin functional activity by neuroactive drugs is discussed.

[Phosphorylation of HMG proteins during change in the chromatin transcription activity of neuronal and glial nuclei of the rat brain]. / Fosforilirovanie belkov HMB pri izmenenii transkriptsionnoi aktivnosti khromatina neironal'nykh i glial'nykh iader golovnogo mozga krys.

The level of cAMP-independent phosphorylation of chromatin, the amount of weakly bound nonhistone proteins and the degree of their phosphorylation in neuronal cells are more significant than the corresponding parameters in glial cells. The mnemic stimulator ethylnorantipheine (ethymizol) increases the level of neuronal chromatin phosphorylation. In contrast, alkylnorantipheine aggravating mnemic processes decreases the phosphorylation. No effect of antipheines on phosphorylation of weakly bound proteins of glial chromatin was found. It is known that whereas ethymizol activates only neuronal, but not glial chromatin transcription, allylnorantipheine inhibits neuronal transcription. The activities of purified casein kinases N I and N II are much higher in neuronal chromatin than in glia. Antipheines have no effect on the activity of casein kinase N I but selectively modulate the phosphorylation of HMG 14 by casein kinase N II of neuronal and glial chromatin. The role of phosphorylated forms of HMG proteins and casein kinases of chromatin in regulation of the genome activity is discussed.

Role of autophosphorylation in regulation of protein kinase CK2 from rat neuronal chromatin.

The regulation of rat brain cortex protein kinase CK2 (casein kinase 2) by autophosphorylation has been investigated. Purified CK2 from rat neuronal chromatin is composed of two regulatory (beta) and two catalytic (alpha and/or alpha;) subunits. The molecular masses of the subunits--43 (alpha), 39 (alpha;), and 25 kD (beta)--were similar to those of typical CK2 subunits. A significant amount of alpha;-subunit occurred in neuronal chromatin; the molar ratios for the subunits were 1.1:0.9:1.9. Pharmacological probes (derivatives of 4,5-di(N-methylcarbamoyl)-1-alkyl-imidazole) were used to study autophosphorylation of separate subunits. These compounds have different effects on neuronal chromatin CK2, transcription, and neurological memory. Changes in the autophosphorylation of the CK2 subunits were found with the help of these compounds. Inhibitors of transcription decreased the beta-subunit autophosphorylation. Stimulators of transcription increased the beta-subunit autophosphorylation and promoted the autophosphorylation of the alpha; (but not the alpha) subunit. The beta-subunit autophosphorylation led to reduction in phosphorylation of nonhistone HMG 14 protein that has been shown to be a physiological substrate of CK2. The autophosphorylation of the alpha;-subunit raised the CK2 activity with HMG 14. The question of functional distinctions between the alpha;- and alpha-subunits of chromatin CK2 in differentiated neurons is discussed.

[Isolation and study of cAMP-independent chromatin protein kinases from brain cells]. / Vydelenie i issledovanie cAMP-nezavisimykh proteinkinaz khromatina iz kletok golovnogo mozga.

Two cAMP-independent protein kinases were purified from rat brain neuron chromatin by using extraction with ammonium sulfate with subsequent chromatography on DEAE-Sephadex A-25 and Sephadex G-150. These enzymes were identified as casein kinases NI and NII, respectively. The molecular masses of the proteins as determined by gel filtration are 4500 and 130 Da. Casein kinase NII utilizes ATP (Km = 7.5 mM) and GTP (Km = 8.5 mM) as substrates, while casein kinase NI utilizes only ATP (Km = 6 mM). The activities of the both enzymes are inhibited by Mn2+ and Ca2+, while heparin (1 microgram/ml) inhibits only casein kinase NII. The memory stimulator ethymizol (ethylnorantipheine) increases the activity of casein kinase NII only when brain proteins extracted by 0.35 M NaCl or rat liver HMG-proteins are used as reaction substrates. This substance has no effect on the phosphorylation of casein and histone HI. The role of casein kinase NII of neuronal chromatin in the realization of stimulatory effects of physiologically active substances on RNA synthesis is discussed.