[Influence of ionizing radiation on enzymatic activity and state of nucleus-nucleolar apparatus in rat hepatocytes].

The effects of a single exposure of rats to the whole-body roentgen irradiation at the doses of 3.5 Gy and 4.5 Gy on the activity of creatine kinase, purine nucleoside phosphorylase, alanine aminotransferase, aspartate aminotransferase, as well as on the state of the nuclear-nucleolar apparatus in rat hepatocytes on the 6th and 13th days after radiation exposure have been studied. Irradiation at the above doses induced changes in the levels of enzymatic activity of different values and different directions within the same time periods, as well as oscillating changes in this type of enzymatic activity over time. This demonstrates various radiosensitivity and adaptation abilities of these enzymatic activities. The changes in the enzymatic activity significantly correspond to the changes in the morphometric indices of nuclear-nucleolar apparatus of hepatocytes, as well as the distribution of hepatocytes within the ploidy classes: in particular, stabilization of the enzymatic activity on the 13th day after irradiation correlates with the increased transcriptional activity, which is detectable through the increased number of nucleoli per nucleus and the expanded space of a hepatocyte nucleus. The compensation mechanisms are likely to be targeted at the changes in the functional activity of surviving hepatocytes, rather than at the replacement of the damaged cells by the new ones.

[The reaction of the neuroblastoma cells in the culture on the influence of tretionine and neurotoxine].

Effect of the tretionine (retinoid) and aluminum chloride (neurotoxin) on the growth and differentiation of neuroblastoma cells in culture after their introduction into the medium separately and in combination was studied. The introduction of these substances creates a new information field in the medium, which becomes apparent by the reactions of neuroblastoma found on the populational and cellular levels of its organization. The presence of tretionine stimulates proliferation and induces differentiation of the cells into astrocytes. Aluminum chloride inhibits cell proliferation and enhances the process of their destruction in the monolayer. The variety of the reactions of neuroblastoma cells to the presence of these substances in the medium indicates the existence and functioning of a mechanism that selects from the information introduced only the portion which may contribute to adaptation of neuroblastoma cells to the changed culture conditions.

[Pathology of lymphoid tissue cells infected by African swine fever virus in vitro].

The authors studied the pathology of bone marrow (BM) lymphoid cell from pigs infected by African swine fever virus (ASFV) in vitro. Monocytes were shown to be primarily afflicted in unstimulated BM culture. These cells disappeared completely 72 hours after infection. Just 24 hours following ASFV infection, there were atypical lymphocytes amounting to 12% of the general lymphoid population at hour 72 after inoculation.The area and perimeter of minor, middle, and large lymphocytes tended to reduce during both BM cell cultivation and inoculation. Lymphoblasts and monocytes were generally triploid in both the control and test groups, but among them there were diploid, triploid, and tetraploid cells. Cytophotometric assay revealed that the amount of nuclear DNA significantly increased in BM lymphoblasts and monocytes in the early stages of ASFV infection (within 24 hours). This effect was also rather pronounced in the lymphoblasts in the later stages (at hour 72).

[The influence of solcoseryl on population and cellular parameters of HeLa and RD cultures].

Changes of population and cellular parameters of HeLa and RD cultures after introducing of solcoseryl in culture medium were studied by methods of scanning cytophotometry and cytomorphometry. Monolayer density, proliferation activity, the number of dead cells in a monolayer, the number of nucleoli in nuclei and distribution of cells in the populations by this parameter, RNA and DNA masses in nuclei and nucleoli, total volumes and surface areas of the nuclei and nucleoli were determined. It has been shown that solcoseryl differently affects the cultures both on population and on cellular levels of their organization. The results of multi-parametric analysis of the influence of solseryl on the cultures allow considering it as a biologically active compound with the features typical for cell and cell population growth regulating factors.

[Multiparametric study and cluster-analysis of the transformed cultures of human cells].

The cells of nepatocarcinoma (HEp-G2), adenocarcinoma of large intestine (Caco-2), embryonal kidney (HEK-293), neuroblastoma (SH-SY5Y), rabdomyosarcoma (RD), and larynx cancer (Hep-2) were studied by the methods of scanning cytophotometry, cytochemistry and cytomorphometry during 96 h of cultivation. The density of monolayers, proliferation activity, the number of dead cells, DNA content in the nuclei and distribution of the cells in the population by this parameter, total DNA content in the nucleoli (circumnucleolar chromatin), the number of nucleoli in the nuclei, distribution of cells by their number, the volume and area of the nucleus surface, total volume and area of the nucleoli surface were determined. The data obtained were used in the treelike cluster analysis of the cultures by Pierson correlation. As a result, the SH-SY5Y culture was put in a separate cluster, while Caco-2, HEp-G2, HEK-293, Hep-2 and RD cultures were placed in the tree of another cluster. The least transformed culture of neuroblastoma (SH-SY5Y) had no relationship with other cultures, which showed various rate of similarity. The cultured HEK-293, Hep-2 and RD appeared to be close to each other by all parameters. The parameters studied are of different significance for the formation of general pattern of the cell cultures. The greatest "weight" is carried by the parameters, which characterize the population as whole: the density of the monolayer, mitotic coefficient and the number of dead cells. They are followed by the content of DNA in the nuclei, the total area of the nucleoli surface, and ratios of DNA content in the nucleoli to DNA content in the nucleus and of total surface of the nucleoli to the surface of the nuclei. Other parameters are not so significant.

[Human large intestine adenocarcinoma cells (CaCo-2) in the process of cultivation].

Using cytomorphometry and cytophotometry cells of human large intestine adenocarcinoma (CaCo-2) were studied under condition of a 10 day cultivation. A reverse dependence was established between proliferative activity and monolayer density. The increase of the latter inhibits proliferation and promotes the formation of islets of polymorph cells. 2c-cells could be seen only at the beginning of culture growth; a larger part of cells polyploidized by cell blocking in G2-phase. These cells do not divide, which is testified by the absence of 2c-cells, but some part of 4c-cells start the next cycle, accumulates 8c-DNA and then divides, replenishing the 4c-cells population. In the process of cultivation, we observed an increase in the number and total volume of nucleoli in the nuclei, and a rise in DNA amount in the peri-nucleolar chromatin. The formation of numerous 4c-cells with multi-nucleolar nuclei may define an increase of functional activity of CaCo-2 culture as the whole, whereas the formation of separated groups of such cells in the monolayer may denote a possible initiation of their differentiation.

[Nuclei and nucleolar organizing regions in chromosomes of lymphocytes on different stages of periodic disease]. / Povedenie iader i iadryshkoobrazuiushchikh raionov khromosom limfotsitov na raznykh stadiiakh razvitiia periodicheskoi bolezni.

By scanning cytomorphometry a cytological study was first performed on the behavior of nuclei and nucleolar organizing regions (NOR) in chromosomes of peripheral blood lymphocytes of healthy men and of patients with periodic disease (familial Mediterranean fever, FMF) on different stages of development, including its complication with amyloidosis. The volume and total surface of nuclei, the sum total volume and sum total surface of NOR, the mean number of NOR for one nucleus and distribution of nuclei according to NOR number were measured. It is shown that the parameters of nuclei and NOR for patients with FMF on all stages clearly and trustworthy differ from those for healthy men. They are sufficiently informative, can be successfully used in clinical practice and even serve as an early diagnostic test for amyloidosis complication.

[Behavior of HEp-2 cell line cells, their nuclei and nucleoli during cultivation and under the effect of Na-ds-RNA]. / Povedenie kletok perevivaemoi kul'tury HEp-2, ikh iader i iadryshek v protsesse kul'tivirovaniia i pod deistviem Na-ds-RNK.

The behavior of human larynx cancer cells (HEp-2) and of their nuclei and nucleoli during the cultivation without the influence of Na-ds-RNA and after its introduction into the medium was investigated by methods of cytomorphometry and cytophotometry. The density of monolayer (the number of cells on the area unit), percentage of two-nuclear cells, the number of nucleoli in the nuclei, mitotic coefficient, volume and total surface of nuclei and nucleoli have been measured. In addition, the mass of DNA in the nuclei and that of the total RNA and DNA in the nuclei and in each nucleolus was measured. Cells in the culture, not subjected to the influence of Na-ds-RNA, were weakly differentiated, kept active proliferation, and their population contained a small number of two-nuclear elements and a high share of multi-nuclear cell. During cultivation, these indices became even more pronounced, which is typical for the increase in cell malignancy. Under the influence of Na-ds-RNA, the proliferate activity decreases, the number of double-nuclear cells increases, while that of multi-nucleolar cell decreases; also, the share of cells with one- and two-nucleolar nuclei increases. The authors conclude that Na-ds-RNA may have antineoplastic activities, clearly evidenced from its influence on the culture of transformed HEp-2 cells.

[The effect of calcium precipitate of double-stranded RNA on erythropoiesis in rats with acute anemia]. / Vliianie kal'tsievogo pretsipitata dvuspiral'noi RNK na éritropoéz u krys pri ostroi anemii.

On the model of experimental acute anaemia of rats, induced by injection of phenilhydrazine, the influence of calcium precipitate of double-stranded RNA (Ca-ds-RNA), introduced during the crise of anaemia, on the process of erythron restoration, was studied. In the presence of Ca-ds-RNA the number of pro- and erythroblasts in fission increases by 1.5 times, compared with "pure" anaemia situation, and accordingly there is a marked decrease in the share of microcytes, which play an important role in the restoration of cell number, and in the erythron recovery after the crise of anaemia. Less pronounced is the influence of Ca-ds-RNA on the macrocytosis, which keeps its value in spite of the increase in the speed of development and of the number of normocytes after Ca-ds-RNA introduction. The mechanism of Ca-ds-RNA inclusion in the system, which controls the process of erythropoiesis during anaemia, is discussed. A conclusion is drawn that Ca-ds-RNA may directly affect the inductive stage of erythropoiesis, stimulating the formation of competent erythroid cells in population of stem haemopoiethic cells and their proliferation. Further development of bone marrow cells takes place according to the known programme of erythropoiesis whose variations are stimulated by the current conditions of its realization, but not by the presence of Ca-ds-RNA.

[Changes in the kinetics of the hemoglobin, total protein and DNA content in rat erythroid cells in experimental anemia. I. The peripheral blood]. / Izmeneniia v kinetike soderzhaniia gemoglobina, summarnogo belka i DNK v éritroidnykh kletkak krys pri éksperimental'noi anemii. I. Perifericheskaia krov'.

Cytophotometry of rat blood erythroid cells during anaemia, induced by phenylhydrazin (4-8 days from the beginning of injections), revealed that all forms of bone marrow containing haemoglobin were thrown into the blood. On its peak (4th day), the greatest contribution in blood haemoglobinization (50%) is made by microcytes. From the 5th day and up to the end of the restoration period the important role in this process is played by macrocytes. From the 6th day the role of normocytes increases, whose contribution by 8th day reaches 70% of the whole haemoglobin amount in blood. In contrary to anaemizated birds, whose erythroid cells ripen in blood, in rats all the transformations of erythron during anaemia are accomplished in bone marrow.

[Changes in the kinetics of the hemoglobin, total protein and DNA content in rat erythroid cells in experimental anemia. II. The bone marrow]. / Izmeneniia v kinetike soderzhaniia gemoglobina, summarnogo belka i DNK v éritroidnykh kletkakh krys pri éksperimental'noi anemii. II. Kostnyi mozg.

Cytophotometry and population analysis of rat bone marrow erythroid cells, during anaemia induced by phenylhydrazin (4-8 days after the beginning of infections), revealed that in the period of restoration of erythron after the rats acute anaemia in the bone marrow blood formation the reciprocally complementary processes of microcytosis, macrocytosis and normal erythropoiesis occur. These processes are based on the normally functioning regulation mechanisms and on the reserve mechanisms, including DNA hyper-replication, which are turned on in the extreme conditions. The various paths of the bone marrow erythroid cells development during anaemia, the means and rates of the micro- and macrocytes formation and part of each line of the erythrocyte development in the restoration of blood cells quantity and haemoglobin homeostasis are analysed. It is suggested that mechanisms of the reserve erythropoiesis, activated during the acute anaemia of adult animals, could function also during the process of normal primary erythropoiesis of embryos.

[Changes in the population composition and the rates of proliferation and specialization of erythroid cells in rats with experimental anemia]. / Izmeneniia sostava populiatsii, tempov proliferatsii i spetsializatsii kletok éritroidnogo riada krys pri éksperimental'noi anemii.

Cytomorphologic, morphometric and population analyses have revealed that during acute anemia, induced by phenylhydrazin, the proliferative activity of erythroblasts increases, an abrupt change in the structures of erythroid populations of both bone marrow and peripheral blood is observed, together with the increase in the rates of their specialization. Also, the ratio of the stem cells which differentiate by the erythroid pathway is increased, and the macro- and microcytosis mechanisms, which are essentially salient in normal conditions, are switched on. The roles of macro- and microcytes in the restoration of the normal amount of erythroid cells and haemoglobin contents in the blood are discussed.

[DNA synthesis and content in the nuclei of epidermal cells of mice during their differentiation and specialization]. / Sintez i soderzhanie DNK v iadrakh kletok épidermisa kozhi myshei v protsesse ikh differentsirovki i spetsializatsii.

Synthesis and content of DNA in the nuclei of differentiating cells of mouse skin epidermis was studied by using cytomorphometric, autoradiographic and cytophotometric methods. It has been shown that the cells of the keratinoid series divide only in the basal layer and contain 2-4c DNA. Keratinocytes of the thorny layer are mostly tetraploid, 2c cells are lacking. H4c and 8c cells comprise 12% of the population. In the keratinocytes of the granular layer DNA content is somewhat lower due to nuclei break down and conversion of cells into anucleate scale. Part of the melanocytes of the basal layer also contain 4c DNA. Highly specialized element of the basal layer Merkel and Langerhans cells are polyploid. Conclusion is drawn that DNA hyper-replication by multiplication of the whole genome is part of the development program of the population.

[Determination of the heparin content in basophilic granulocytes by cytophotometry]. / Opredeleneie soderzhaniia geparina v bazofil'nykh granulotsitakh s pomoshch'iu tsitofotometrii.

A method for estimating heparin content in basophilic leucocytes by means of cytophotometry of alcian blue--heparin complex is proposed. Its application for measuring heparin amounts in basophils of healthy donors and of patients with allergies is demonstrated.

[In vitro cell culture of the Ararat cochineal insect, Porphyrophora hamelii Brandt]. / Kul'tura kletok Araratskoi koshenili Porphyrophora hamelii Brandt in vitro.

The possibility of cultivating the Ararat cochineal cells in the nutrient media developed for cell cultures of the other insect species was investigated. The initial cochineal cell culture was obtained in suspension. The cell culture on the substrate (glass) dies within 7 days. 2 peaks of proliferative activity was observed in the suspension culture. The increase of cell ploidy and their death were found between these two peaks. The small cells survived and kept their capacity for proliferation. The hemolymph did not stimulate cell proliferation but enabled their better attachment to the substrate. The use of different media did not reveal marked differences in cell behaviour. The pigment granules disappeared during the first week of cultivation.