[Blockade of proteasome activity disturbs the rhythm of synthesis of the protein marker of direct cell-cell interactions].

The effect of inhibition of proteasome activity on direct cell-cell interactions in primary hepatocyte cultures was studied. The circahoralian rhythm of protein synthesis was a marker of cell-cell communication. The addition of the proteasome inhibitor MG132 at doses of 10 or 20 µM to the medium with hepatocyte cultures for 19 h resulted in a significant reduction in the total pool of 3H-leucine in cells. The incorporation of leucine into proteins changed slightly or negligibly, whereas the content of free labeled leucine in hepatocytes decreased. The rhythm of protein synthesis was distorted compared to the control. The rhythm was restored by external organizers, such as gangliosides and melatonin, as well as by enhancing the activity of protein kinases--the key factor in the organization of the rhythm of protein synthesis. A short-term (3-h) exposure to MG132 did not change the pool of leucine, but the rhythm of protein synthesis was also disturbed. Thus, protein catabolism affects cell-cell interactions organizing the rhythm of protein synthesis. Another factor of the downregulation of the rhythm of protein synthesis, the secretion of proteins from the hepatocytes in vivo, which was shown in vivo in many studies, was also revealed in our study when measuring the content of proteins stained with Coomassie Brilliant Blue G250 in the medium with hepatocyte cultures.

[Effect of cell microenvironment on cell functions associated with tumour promotion and progression].

To study the tumour-promotion activity of cell environment the transformed embryonic rat fibroblasts (clone CL-1-1) were transfect to immunodeficient mice then the cells of the formed tumour were cultivated (clone CL-1-1). The cells before and after transplantation were compared by morphology, proliferation activity and gap junction intracellular communications. The clone CL-1 cells proliferated much faster than clone CL-1 cells. The CL-1-1 cells had changed morphology structure and unlike CL-1 the contract inhibition was absent in CL-1-1. The number of CL-1 cells in phase G1 was significantly greater than that of CL-1-1 cells, while the number of CL-1-1 cells in G2/M phases was much more the number of CL-1 cells. The activity of gap junction intercellular communications in both cell types was near the same. It was concluded that cell microenvironment act as a tumour-promoter and tumour progression factor in the case of cell transplantation to immunodeficient mice.

[Melatonin injected into rat efficiently synchronizes the rhythm of protein synthesis in primary hepatocyte cultures isolated from this rat].

Melatonin injected intraperitoneally into rat synchronizes the ultradian rhythm of protein synthesis after 100 min in primary hepatocyte cultures isolated from this rat, which are studied after 1 or 2 days. The effective synchronization concentrations of melatonin--0.01-0.02 microg per kg of rat weight--are three orders lower than melatonin doses used in clinical practice in human treatment.

[Activation of reconstructive processes in rat tissues under the action of radiofrequency current with a periodic impulse mode of modulation].

The action of a current in the radio frequency range with a periodic impulse mode of modulation on the activation of recovery processes in the skin and skeletal muscles has been studied. The action of a radio frequency current with a power of 1 W, as opposed to that of the weaker action (0.1 W) and stronger (4 W) action, leads to the activation of recovery processes in the skin and skeletal muscles. Recovery processes are manifested in the increase in proliferation and activation of angiogenesis in the skin, and also in formation of new muscle fibers. Recovery processes in muscles are accompanied by activation and migration of satellite cells of muscle tissue in the zone of action of the radio frequency current.

[Melatonin promotes and synchronizes protein synthesis in hepatocyte culture from old rats].

The effect of 1 to 1000 nM melatonin was studied on daily cultures of rat hepatocytes on slides in serum-free medium. The minimum melatonin concentration (1 nM) proved to synchronize protein synthesis in asynchronous sparse cultures of hepatocytes from rats of different age, and a circahoralian rhythm of protein synthesis was revealed in them. In dense weekly synchronous hepatocytes from old rats (2.5-years-old with the weight of about 600 g), melatonin improved cell synchronization to the level of young animals. Melatonin treatment increased the mean rate of protein synthesis in rats of different age.

[The role of the vascular factor in the reorganization of water metabolism in denervated liver after bacterial endotoxin poisoning].

Intoxication with bacterial lipopolysaccharide (endotoxin) is accompanied by considerable rearrangements in the systems of blood microcirculation and water metabolism of the liver. These rearrangements are manifested as increased sinusoid area, changed total area of the cytoplasm and nuclei as well as the nucleocytoplasmic ratio in hepatocytes, increased content of total water in the organ, and changed magnetic relaxation properties (spin-lattice and spin-spin relaxation times). Preliminary parasympathetic denervation of the liver (vagotomy) changes the pattern of the organ response to bacterial endotoxin poisoning as indicated by the kinetics of studied morphological and biophysical parameters.

[Changes in the morphofunctional status of the liver in rats with acute tetrachloromethane poisoning and their correction with antihypoxants, antioxidants and actoprotectors]. / Izmeneniia morfofunktsional'nogo sostoianiia pecheni krys pri ostroi intoksikatsii tetrakhlormetanom i ikh korrektsiia antigipoksantami, antioksidantami i aktoprotektorami.

The acute tetrachlormethane intoxication leads to structural and metabolic alterations in the rat liver. Morphological changes include centrolobular necroses and blood stasis in dilated sinusoidal capillaries. Metabolic changes are manifested by an increase in the water content and time of the spin-lattice (T1) and spin-spin (T2) relaxation (this indicates a lesser degree of water structuralization and its enhanced lability), and distortion of the correlation between T1 and T2. The prophylactic administration of one of the antihypoxants, antioxidants or actoprotectors normalizes the morphofunctional condition of the liver. According to the degree of the therapeutic efficacy, the examined preparations form the following series: dibunol > gamma-sodium hydroxybutyrate > tomerzol.

[Changes in the serum albumin system during toxic syndrome and their pharmacologic correction]. / Izmeneniia sistemy syvorotochnykh al'buminov pri toksicheskom sindrome i ikh farmakologicheskaia korrektsiia.

In connection with the important role of serum albumins in pathogenesis and sanogenesis of numerous toxic states, we examined binding capacity of these proteins and conditions of their binding sites after acute poisoning with tetrachloromethane (TCM) and administration of antihypoxic agents (sodium gamma-hydroxybutirate), antioxidants (Dibunol), and actoprotector (Tomersol) to rats. We demonstrated that tetrachloromethane intoxication (3.2 g/kg over 24 h) was accompanied by a certain decrease (by 13.8%) in the total blood serum level of albumins and a tendency to a decrease in the value of the binding constant of negatively charged fluorescent probe 1-(phenylamino)-8-sulfonaphthalene. Under these conditions, the mean number of probe binding sites per albumin molecule increases, and as a result, the total concentration of albumin binding sites in the serum remains virtually unchanged. We found that accessibility of the probe to a quenching agent (potassium nitrate) increases in the protein--probe complex under intoxication conditions, suggesting that the type of interaction between the protein and the fluorescent probe changes as well. Therapeutic/prophylactic administration of an antioxidant, antihypoxic agent, or actoprotector leads to an increase in the level of albumin in the serum (Tomersol), partial normalization of its binding properties (binding constant in the case of sodium gamma-oxybutirate, mean number of binding sites per molecule for Dibunol and Tomersol), and the state of binding sites (sodium gamma-oxybutirate, Dibunol).

[Morpho-biochemical study of the liver and serum albumins during tetrachloromethane intoxication and administration of antihypoxants, antioxidants, and actoprotectors]. / Morfobiokhimicheskoe issledovanie sostoianiia pecheni i sistemy syvorotochnykh al'buminov v usloviiakh intoksikatsii tetrakhlorometanom i vvedeniia antigipoksantov, antioksidantov i aktoprotektorov.

Organism intoxication with tetrachloromethane induces a complex of transformations in the liver and serum albumins system manifested in the liver as necroses in the central lobe, dilatation of sinus capillaries and blood stasis in them, increased content of total water, and changed magneto-relaxation properties indicating decreased structurization of water. The system of serum albumins shows decreased total concentration of albumins in the serum, decreased constant of 1-(phenylamino)-8-sulfonaphthalene probe binding, increased mean number of probe binding per albumin molecule, and increased accessibility of probe in the protein-probe complex for the quencher indicating changed type of interaction between the protein and fluorescent probe. Introduction of a antihypoxant, antioxidant, or actoprotector has a positive effect on the condition of the liver and serum albumins system: The studied indices are nearly completely normalized.

[The rhythm of protein synthesis in denervated rat liver]. / Ritm sinteza belka v denervirovannoi pecheni krysy.

Using the radiochemical method, we found a circahoralian rhythm of protein synthesis in slices of rat liver two weeks after transection of the vagus nerve anterior stem or bilateral subphrenic vagotomy with simultaneous transection of the celiac plexus hepatic branches. The rhythm was similar to those found in slices of normal liver or in a monolayer culture of hepatocytes. The incorporation of leucine in proteins and the pool of free leucine were similar in the denervated and normal liver. The self-synchronization of individual cell oscillations in protein synthesis intensity was shown earlier in experiments performed on cell cultures. A study of protein synthesis in the denervated liver suggests that cell interactions are also the main mechanism underlying the synchronization of oscillating cellular functions in the organ in situ.

[Changes in reactivity of the contractile system of the small intestine caused by disturbed parasympathetic innervation]. / Izmenenie reaktivnosti sokratitel'nogo apparata tonkoi kishki v usloviiakh narushennoi parasimpaticheskoi innervatsii.

We studied the role of parasympathetic division of vegetative nervous system in regulation of the intestine activity in rats subjected to bilateral subdiaphragmal vagotomy, the response of the neuromuscular apparatus of the small intestine to stimulation by aceclydine taken as an example. Intravenous injection of aceclydine (0.5 mg/kg) into intact animals is accompanied by characteristics changes in the electrical and contractile activities of the intestine, which reflect its enhanced motor activity. Vagotomy leads to changes of response of the intestine to pharmacological stimulation, most expressed within 3, 7, and 30 days. It was proposed that distortion of reactive properties of the intestine contractile system under these conditions is related to functional changes in rhythm-setting neurons of intramural ganglia and M-cholinoreceptors of neuromuscular synapses.

[The role of the parasympathetic nervous system in regulating the kinetics of the regional blood circulation in the digestive system organs]. / Rol' parasimpaticheskoi nervnoi sistemy v reguliatsii kinetiki regionarnogo krovoobrashcheniia v organakh pishchevaritel'noi sistemy.

Cranio-bulbar part of the parasympathetic nervous system proved to be important for regulation of the blood flow distribution between the organs of the gastroduodenal complex (liver, stomach, and small intestine), rate of the local blood flow and its kinetics. Investigation of the blood supply of the above mentioned organs various periods (1-60 days) after bilateral subdiaphragmatic truncal vagotomy has demonstrated the most pronounced transformations in macro- and micro-hemodynamics in the 14-30 days period. Time pattern of these changes has one maximum in liver and stomach (14-30 days) and two maxima (7 and 30 days) in small intestine.

[Experimental validation of the use of betamecil in ophthalmology]. / Eksperiemntal'noe obosnovanie primeneniia betametsila v oftal'mologii.

Wound-healing activity of 2% betamicil ointment in comparison with the traditional stimulants of corneal repair regeneration (methyluracyl and solcoseril) was studied in experiments. The rate of epithelialization of a standard trephination wound in rabbit cornea (28 animals), mitotic activity of the anterior corneal epithelium, and strength of the regenerated tissue were assessed after use of different stimulants and in control. A maximal positive effect of local betamicil (2% ointment) was observed: the wounds epithelialized 57% sooner than in control, but there was no reliable difference from solcoseril; the regenerate of linear corneal wound was 1.5 times stronger. Use of this ointment in ophthalmology is validated.

[Use of antihypoxants and antioxidants for pharmaco-correction of disorders of water metabolism in the liver and small intestine after vagotomy]. / Primenenie antigipoksantov i antioksidantov dlia farmakokorrektsii narushenii metabolizma vody v pecheni i tonkoi kishke posle vagotomii.

Bilateral subdiaphragmatic vagotomy in rats leads to hepatic and small intestinal water metabolic disorders appeared as higher levels of total water, longer spin-grid (T1) and spin-spin (T2) relaxation and impaired correlation between T1 and T2. The time course of changes in water metabolism in the liver is one-sided (maximum after 7 days) at the same time in the small intestine is two-sided (maximum after 7 and 30 days). The antihypoxant tomerzole and the antioxidant dibunol produce a partial normalizing effect on water metabolism in the studied viscera of the animals vagotomized (at day 30), the highest efficiency was demonstrated by dibunol whose pharmacological activity is the greatest in the small intestine.

[The role of the nervous factor in regulation of oxygen supply to the small intestine]. / Znachenie nervnogo faktora v reguliatsii kislorodnogo snabzheniia tonkoi kishki.

The experimental model of rat bilateral subdiaphragmatic stem vagotomy was used to study the role of parasympathetic nervous system in regulation of circulation and oxygen supply to the small intestine. Cut of the vagus nerves is shown to cause redistribution of the blood flow between gastroduodenal organs, to slow down local circulation, to reduce oxygen tension in the muscular lining of the jejunum, to raise water content in the wall and change magnetorelaxational characteristics. Morphologically this manifested in altered arteriolar and capillary configuration, dilation of the capacity microvessels, dystrophia of microvessel wall, red cell aggregation in capillaries and venules, defective permeability of the vascular wall. The above indices changed most 7 and 30 days after vagotomy. It is evident that impaired vagus innervation produces secondary circulation hypoxia in the small intestine.

[Condition of erythrocytes in intact and vagotomized rats of various age after massive hemorrhage].

Sharp massive hemorrhage (30-35 % of total amount of blood) in rats are accompanied by natural changes of morpho-functional characteristics of erythrocytes: diameter, the area, polarization, the factor of the form, integrated and specific optical density. Thus, in certain terms, after hemorrhage at young initially intact animals average diameter, the area, integrated optical density actually increase as well as the value of polarization and the factor of the form of erythrocytes growth. In vagotomized rats of the same age group, in reply to hemorrhage, the increase in value of the factor of the form only is obvious. In initially intact old animals in these conditions, an increase of integrated and specific optical density and of the factor of the form also takes place. In old vagotomized rats, the average diameter and the erythrocyte area accrue. Thus, the character and dynamics of response of erythrone in initially intact and vagotomized (through 14 days after operation) animals essentially differ and have certain features dependent on age. Thus, in young initially intact rats, the most expressed changes are marked in 3-10 hours whereas in vagotomized animals of the same age group - in 3 and 96 hours after hemorrhage. In old initially intact rats, the greatest deviations of the investigated parameters are observed in 3 and 96 hours, in the vagotomized animals - in 3 and 24 hours.

[Complex analysis of the state of sinusoid vessels of the liver after disturbance of innervation]. / Kompleksnyi analiz sostoianiia sinusoidnykh sosudov pecheni posle narusheniia innervatsii.

By means of light, electron microscopy and morphometry, the state of the sinusoid vessels and cells of their lining has been investigated in 7, 14, 30, 60 and 90 days after bilateral subdiaphragmal vagotomy. In the days mentioned the cross section area of the sinusoid vessels and amount of active labelled Kupffer cells significantly increase (except day 90 ). By the end of the 3d month these parameters decrease, but they do not reach the control level. Ultrastructure of the endothelial cells and Kupffer cells is changed, phagocytic activity of the latter is increased. The greatest destructive changes take place in Kupffer cells, especially during first two months. By the end of the 3d month there is certain tendency to normalization.

[Characteristics of the regeneration process in the liver deprived of polyploid hepatocytes]. / Osobennosti regeneratsionnogo protsessa v pecheni, ne soderzhashchei poliploidnykh gepatotsitov.

By applying autoradiography with 3H-thymidine hepatocytes of young adult guinea pigs were found to enter the DNA-synthesis phase 25 hours after partial hepatectomy. Peaks of nuclei labeling were revealed 30, 45, and 60 hours after the operation. By calculating mitotic figures on squash preparations two waves of mitosis were found (in 40 and 55 hours) after hepatectomy. Cytophotometric study of DNA content showed practically all mononuclear and binuclear hepatocytes to contain diploid nuclei in 3 and 5 days after the operation. By the end of the 7th regeneration day there were revealed 6% mononuclear tetraploid cells. The percentage of binuclear cells during the period of regeneration under study decreased from 16 to 8%. A conclusion was drawn that the main cytologicalmechanism of the guinea pig liver regeneration was full-fledged mitosis ending by cell division.

[Effects of ethidium bromide on the viscosity of chromatin secreted by rat liver in normal state and after denervation]. / Vliianie bromistogo étidiia na viazkost' khromatina, vydelennogo iz pecheni krys v norme i posle denervatsii.

The paper studies structural state of chromatin secreted from the rat's liver. For this purpose the authors studied the dependence of relative viscosity of chromatin preparations secreted from the liver of rats and three groups of animals being in different terms of postdenervation process.

[Evaluation of the functional state of the hepatocyte monooxygenase system in rats with damaged innervation of the liver]. / Otsenka funktsional'nogo sostoianiia monooksigenaznoi sistemy gepatotsitov krys pri narushenii innervatsii pecheni.

Cytochrome P-450 content measured spectrophotometrically 14 and 25 days after bilateral subdiaphragmal vagotomy in hepatocyte microsomes of inbred male rats was considerably reduced. The rate of aminopyrine demethylation and oxygen consumption decreased, while the duration of hexenal-induced sleep was prolonged. Monoxygenase responsible for p-hydroxylation of aniline proved relatively resistant to vagotomy.