ISOLATION AND COMPERATIVE STUDY OF THE GROWTH INHIBITING THERMOSTABLE PROTEIN COMPLEX FROM THE BONE MARROW OF THE ADULT MICE.

The growth inhibiting thermostable protein complex (TPC) from the bone marrow cells of adult mice was isolated and partially characterized. A comparative analysis of TPC components was performed by polyacrylamide gel electrophoresis. It has been established that the complex isolated from the bone marrow of adult mice, like complexes obtained from other organs, contains two relatively high-molecular-weight and low-molecular-weight sub fractions of proteins. In addition, minor content of low molecular weight components was detected in bone marrow TPC. It has been established that the thermostable protein complex of adult mice bone marrow cells does not have the ability to inhibit the proliferation of homotypic cells. This may be due to the minor content of active component (low molecular weight subfraction) in the complex and the structural and functional properties of constantly renewable hematopoietic tissue, such as the nature of factors that regulate proliferation, their different cell penetration and the expression of membrane receptors.

PECULIARITIES OF ACTIVATION OF COMPENSATORY-ADAPTIVE PROCESSES IN ADULT RAT LIVER CAUSED BY UNILATERAL NEPHRECTOMY.

In order to determine the general patterns of activation of the inter organ compensatory-adaptive processes, the peculiarities of activation of compensatory processes caused by unilateral nephrectomy in adult rat liver at the initial stage of hepatopathy (first three days after the operation) have been studied. In particular, it has been established that only small but visible abnormalities in the spatial relationship of sinusoids and hepatic plates are revealed morphologically in rat liver histology. The increasing of the functional load caused by changes in histoarchitecture, at these times, does not stimulate hepatocyte proliferation. At the same time, it is revealed that at the initial stage of hepatopathy caused by unilateral nephrectomy, preference is given to polyploidy from compensatory-adaptive processes charachterised to liver. In particular, it has been shown that liver responses mainly by quantitative increases in binucleated cells (2cx2) to the functional load induced by unilateral nephrectomy at an early stage.

THE STUDY OF THE IMMUNE CORRECTIVE PROPERTIES OF GREEK WALNUT (JUGLANS REGIA L.) SEPTA ON THE EXPERIMENTAL MODEL OF LEUKOPENIA.

From the positive effects of the drugs prepared from various parts of walnut described for a wide variety of diseases, their antitumor effect is remarkable. This feature can be used for treatment of leukopenia caused by radiotherapy and/or chemotherapy. Therefore, to study the immunocorrective properties of Greek walnut, the walnut septa were selected, aqueous extract of which has been studied on experimental model of leukopenia in white mice caused by a single injection of cyclophosphamide. The material of the study were the blood and bone marrow smears from intact and tested adult mice stained by Giemsa's dye. The quantity of leukocytes in peripheral blood were determined by the counting chamber under the light microscope with standard protocol. Cloud-point extraction, HPLC analysis and UV-visible spectrophotometry were used to study the composition of the walnut septa extract. It has been established that the Greek walnut septa extract has the correction ability of suppressed myelopoiesis in white mice caused by injection of cyclophosphamide. The blood formula normalization process by the mentioned extract is provided by the fast increasing in number of immature (band neutrophil) and mature neutrophils in the peripheral blood. It was shown that walnut septa extract stimulates the division, differentiation and maturation of blast forms of myeloid as well as lymphoid line in the bone marrow of mice with leukopenia. Cloud-point extraction and liquid-chromatographic analysis revealed the presence of hydrophilic and hydrophobic components in the walnuts septum extract.

COMPARATIVE STUDY OF GROWTH INHIBITORY PROTEIN COMPLEX DERIVED FROM DIFFERENT FORMS OF CHILDREN HEMANGIOMA.

The comparative analysis of the thermostable protein complexes obtained from the capillary and the cavernous hemangioma was carried out. It is shown that the active component (with low molecular weight 12-14 kD) is in minor content in the complexes obtained from capillary, as well as from the cavernous hemangioma. The effect of protein complexes from cavernous hemangiomas on proliferation of the brain and pancreatic cells in growing rats was studied. It was established that the thermostable protein complex from cavernous hemangiomas does not have the ability to inhibit the proliferation of heterotypic cells due to the minor content of the active ingredient in protein complexes.

Ubiquitin in combination with alcohol stimulates proliferative activity of hepatocytes.

There are several factors involved in aetiology of Alcoholic Liver Disease. The one of the most important role in regulation of intracellular transformations belongs to ubiquitin proteasome system that is comparably well studied, but less is known about the functions of extracellular ubiquitin. The goal of presented work is to study the effect of extracellular ubiquitin on partially hepatectomized alcoholic liver regeneration. Experiments were carried out on white female rats. Proliferative activity of hepatocytes was determined by colchicine mitotic index and immunohistochemical staining on Ki67. We have shown that in vivo injected extracellular ubiquitin stimulates proliferative activity of hepatocytes in partially hepatectomized alcoholic liver whereas in non-alcoholic rats hepatocytes proliferative activity was decreased. The main attraction of the experiment is the combination of alcohol and ubiquitin, as without alcohol ubiquitin inhibits proliferative activity of hepatocytes and blood cells. Further investigation of its action will provide new insights into possible pathway of ubiquitilation involved in stimulation of proliferative activity in alcoholic liver.

Pancreatic cell proteome - qualitative characterization and function.

The macromolecular protein complex form adult white rat pancreas has been obtained and partially characterized. The most important distinguishing characteristics of the complex is the thermostability of its components. Using polyacrylamid gel by electrophoresis and chromatography of hydrophobic interaction has been established, that the components with relatively high (45-60 kD) molecular weight, are hydrophobic, while the low molecular weight components (11-12 kD) corresponds to hydrophilic proteins according to its column retention time. Participation of TPC in the regulation of the homotypic cell growth was determined. It is shown that the pancreatic TPC through inhibition of transcription decreases cell mitotic activity in growing rats. It is also shown that this complex is involved in pancreas regeneration processes. On the basis of the qualitative characteristics and cell growth regulatory function of this macromolecular protein complex described by us, it can be considered as a pancreatic cell proteome.

Influence of halothane narcosis on a change of the number of GABA-positive cells in the hippocampus of adult rats with the midasolam premedication.

The most difficult questions in pediatric anesthesiology still remain relevant at surgeries to correct congenital maxilla-facial pathologies, accompanied by severe respiratory distress, speech and auditory sensation. For a long time the anesthetics were widely used among the neonatal and young children with extreme care due to their indicated suppressive effects on the immature organ systems. According to this fact, the choice of anesthesia is a matter of extremely importance. In addition, the study of molecular mechanisms indicating to the adverse effects induced by application of anesthesia will make it possible to develop therapeutic strategies for prevention of postoperative complications. The purpose of present study is to investigate a change of the quantity of GABA positive cells in the hippocampus of white adult rats under the conditions of halothane narcosis and with the premedication by midazolam. Subjects of a study - 30 adult white rats with the weight of 130-140 g. Animals were subdivided into three groups. I group - intact animals; II group - sham-operated rats under the conditions of halothane narcosis; III group - sham-operated animals, premedicated by midazolam, under the halothane narcosis. A change in the quantity of GABA positive cells was determined by the immunohistochemical method (Rabbit Anti-GABA). All data were processed by the method of standard variation statistics. The authenticity of results was evaluated according to the Student's criterion within limits of 95-99% reliability. The results of conducted by us studies revealed, that during the application of halothane narcosis causes an increase in the GABA positive cells in the fields CA1 on hippocampus of adult white rats. In 24 hours after sham operation, at the condition of application of halothane narcosis a quantity of GABA positive cells raises in the CA3 field of hippocampus in II group animals as well. Moreover, in the CA1 field correlation between GAD65/67 positive and GABA positive cells is revealed which was not observed at later stages. The study of this question is the purpose of our next investigations.

In vivo investigation of extracellular ubiquitin effect on liver histoarchitectonics.

Ubiquitin is a small regulatory protein of living cells. It affects almost all cellular processes. Investigation of ubiquitin is very important for discovering the new therapeutics for different heavy human diseases. The main goal of presented work was to reveal effect of extracellular ubiquitin on liver histoarchitectonic of mature and immature white rats. The obtained data indicate the inefficiency of used doses--200 µg/ml and 500 µg/ml--of extracellular ubiquitin single and multiple in vivo injections to evident changes of histoarchitectonics of immature and mature white rat liver. Further investigation of in vivo injection effects on histoarchitectonics of intact and damaged liver and hepatocyte cell cycle regulation by immunological assays seems to be actual and interesting.

The positive effects of Midazolam on functional activity of white rat brain cells in conditions of halothane anesthesia.

For the elimination of postoperative complications, which are evident in infants with congenital palatine and upper lip cleft after halothane anesthesia during standard premedication (Atropine, Dimedrol) the usage of agents of benzodiazepine group with anti-hypoxic effects is advised. The latter may modulate the blocked ion channels by neuromediators, which are activated by GABA and GABA receptor function. The neuro-protective ability of halothane is demonstrated. To reveal the mechanisms of positive effects using benzodiazepine group for premedication we have investigated the effects of midazolam of brain cell functional activity of experimental animals (white rats) in conditions of halothane anesthesia. For the estimation of white rat brain cells functional activity the nuclear transcriptional activity was studied (based on the intensity of [14C]-UTF inclusion), also the glutamic acid decarboxilaze activity (GAD65/67) using immunohistochemistry. It is estimated that halothane inhibits the transcription in rat brain cells. During midazolam premedication the halothane inhibitory effect on RNA synthesis is not revealed. After an hour of pseudo-operation halothane also induces proved decrease of quantity of GAD65/67 positive cells in CA3 hippocampal field. At the same time the quantity of similar cells are increased in CA1 field. The increase of GAD65/67 positive cells in CA1 is more evident during midazolam premedication. Based on the data received we can conclude that the positive effect of midazolam results in increase of GAD65/67 positive cells in CA1 hippocampal field.

[Kinetic features of a white rat hepatocyte population under hormonal disbalance before and after partial hepatectomy].

Bilateral adrenalectomy, followed in 4 days by a partial hepatectomy, was performed using white rats weighing as much as 120-140 g. Under hormonal disbalance caused by bilateral adrenalectomy, the number of polyploid (4c, 4c x 2, and 8c) hepatocytes significantly increased, compared to that in non-operated control rats. Six hours after a partial hepatectomy, the share of highly ploid hepatocytes falls, being accompanied by a 9-fold increase in mitotic index. It is supposed that under hormonal disbalance condition, a partial hepatectomy may induce "early" mitoses in hepatocytes blocked in G2-phase of the cell cycle.

[Regeneration of white rat liver during cholestasis]. / Regeneratsiia pecheni belykh krys pri kholestaze.

Cholestasis inhibits expression of early response genes in rat hepatocytes stimulated to proliferation and restricts the process related to reparative regeneration. The occlusion of general bile duct induces bile acid accumulation, which suppresses 11-beta-hydroxysteroid-dehydrogenase activity. Hence, the concentration of corticosterone is increased, which suppresses the expression of early response genes in rat hepatocytes. The restoration of liver mass occurs due to proliferation of reserved cells. An unusual mitotic peak of these cells is registered 34 h after the operation.