[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.

[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.

[Recognition of cell patterns and prospects of applying it in analytical microscopy]. / Raspoznavanie obrazov kletok i perspektivy ego ispol'zovaniia v analiticheskoi mikroskopii.

On the base of multiparametric analysis of scanograms, a method of creation of cell patterns and of their recognition in n-dimensional space is suggested. The prospects of its application in the analytic microscopy and in differential diagnostics of diseases are discussed.

[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.

[The morphofunctional changes in the hepatocytes of rats with experimental acute pancreatitis]. / Morfofunktsional'nye izmeneniia gepatotsitov u krys pri éksperimental'nom ostrom pankreatite.

Under experimental acute pancreatitis of rats, certain changes were revealed in the liver histological structure, hepatocyte population composition, intensity of hepatocyte DNA-synthesis, their proliferation, and in hepatocyte glycogen contents. These changes reflect a two-step process of hepatocyte activation. This results well compares with the earlier evidence on changes in pancreatic cell population under acute pancreatitis. The role of hepatocyte protective reactions during regenerative process in pancreas, under acute pancreatitis and its transition into chronic condition, is discussed.

[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.

[Morphofunctional changes in the exocrine pancreatic cells in acute experimental pancreatitis in rats]. / Morfofunktsional'nye izmeneniia ékzokrinnykh pankreatsitov pri ostrom éksperimental'nom pankreatite u krys.

Experimental pancreatitis was induced by cooling the splenetic part of rat pancreas with chlorethyl, and the cells of duodenal area of the pancreas were studied at different stages of pancreatitis using cytomorphometry, cytomorphology and autoradiography. Interlobular and interacinar oedemas were observed at the first hours after treatment. In 24 hours the intracellular oedema of exocrine pancreatic cells (EP) was detected. On day 14 after treatment typical acute edematous pancreatitis developed. The observed changes involve a pathological activation of EP of the duodenal area, a subsequent restoration of the structure of this area, and later a passage of pancreatitis into the chronic form. The usefulness of this model of pancreatitis for quantitative cytochemical studies of EP during pathogenesis and drug treatment is discussed.

[Cytological research on the action of sodium thiosulfate on the process of induced acute pancreatitis in rats]. / Tsitologicheskoe issledovanie deistviia tiosul'fata natriia na protsess indutsirovannogo ostrogo pankreatita u krys.

The influence of sodium thiosulfate (STS) on the process of experimental acute pancreatitis (EAP) in rats was studied by cytomorphology, morphometry, autoradiography and cytophotometry. The influence was shown to vary at different stages of disease development. At the first stage ("primary effect" state) STS leads to the increase in the stability of exocrine pancreacytes (EP) against the toxins and to the decrease in the activity of proteases formed during necrobiosis. This results in the drop of the number of degrading EP and of the degree of inter- and intracellular oedema, and brings about shifts towards the normal values of the nucleus cytoplasm shapes, the nucleus/cytoplasm ratio, the EP population structure and their RNA and protein content. At the second stage STS stimulates DNA synthesis in EP and their proliferation leading to accelerated restoration of the number of viable cells. STS also stimulates the regeneration process hence preventing pancreatitis from passage to its chronic form. The mechanism of STS action of EP functions in normal cells and during pathogenesis is discussed.

[DNA synthesis and content in the macrophage nuclei of normal mice, during carcinogenesis and during the administration of retinoids to the animals]. / Sintez i soderzhanie DNK v iadrakh makrofagov myshei v norme, pri kantserogeneze i pri vvedenii zhivotnym retinoidov.

It has been found that nearly 50% of the lymph node and spleen macrophages (MP) of the CBA line mice contain DNA at levels superior to the diploid value (H2c--H4c in mononuclear MP, and up to H16c among polynuclear ones, the latter comprising 2.5-9.0% of the whole MP population). No DNA synthesis and mitosis were detected by autoradiography, cytophotometry, and cytomorphological analysis. During carcinogenesis the proportion of MP with elevated DNA amounts ("activated MPs") decreases due to their migration to tumours. Also immature MPs (1.6%) appear in the population, which synthesize DNA, but do not divide. Injection of retinoids restores the percentage of MPs with elevated DNA amounts to the levels characteristic of the intact animals, the fraction of DNA-synthetizing cells increasing up to 2.8%. It is proposed that retinoids may accelerate the processes of MP maturation, activation and renewing. A mechanism of cooperative action of MPs and retinoids is discussed in addition to the role of DNA hyper-replication.

[Changes in the content of nucleic acids and total protein in exocrine pancreatocytes and the composition of their population in experimental acute pancreatitis in rats]. / Izmenenie soderzhaniia nukleinovykh kislot i summarnogo belka v ékzokrinnykh pankreatsitakh i sostava ikh populiatsii pri éksperimental'nom ostrom pankreatite u krys.

Using cytophotometry, contents of DNA, RNA and total protein were measured in the rat's exocrine pancreatocytes (EP) in normal conditions and at different stages of pancreatitis induced by cooling the spleen part of the pancreas with chlorethyl. In the duodenal (not damaged) part of the pancreas some drastic changes in the EP ploidy distribution were shown to occur. They led to the formation of a qualitatively new population pattern with 4c and 2c + 2c cells prevailing (more than 60% of the total content), which are by 1.5-3 times more active in RNA and protein synthesis and accumulation than the normal cells. The population structure rearrangement in the EP and their functional activity rise took place at two stages. At the first one the intracellular defense mechanisms of the EP in response to acute necrobiotic processes in the pancreas tissue were activated, at the second one the supracellular mechanisms regulating synthetic processes leading to a rapid adaptation of viable EP to new conditions were switched on.

[Two types of fragmentation of total chromatin by DNAase I--an artifact or reality?]. / Dva tipa fragmentatsii total'nogo khromatina DNKazoi I--artefakt ili real'nost?

The possibility of N+2N repeat (a nucleosomal-type repeat in which the even-numbered peaks dominate) being an artifact has been studied. The repeat results from digestion of chromatin of several rat cells by DNAase I. Endogenous nucleases are not shown to be involved in formation of the repeat. N+2N repeat is also formed during digestion of nuclei isolated from homogeneous lymphocyte populations indicating that the repeat is inherent of chromatin of distinct cells and is not the result of superimposition of different repeats arising from diverse tissue cells. We suppose the N+2N repeat to indicate the existence of the second type of total chromatin differing from the one giving rise to the dinucleosome repeat under the same conditions.

[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.

[The coefficient of the relative position of particles and its use in the quantitative analysis of substance distribution in cells]. / Koéffitsient vzaimoraspolozheniia chastits i ego ispol'zovanie pri kolichestvennom analize raspredeleniia veshchesta v kletkakh.

A new method of objective estimation of material distribution in micro objects is described, namely, computing of the coefficient of mutual location of particles, while the cell image is presented as a digital map. The two methods of estimation of distribution of the material are compared - the coefficient of mutual location and the texture coefficient. Advisability of the parallel use of both the methods is substantiated.

[Total protein content of cells as a factor regulating their proliferation]. / Soderzhanie summarnogo belka v kletkakh kak faktor reguliatsii ikh proliferatsii.

Proliferation kinetics in stimulated stationary cultures of chick embryo fibroblasts was studied using cytophotometric and autoradiographic methods. Part of 4c cells are blocked at G2 when the culture becomes stationary. A fraction is formed among them which fails to divide in response to proliferation stimulus. Such cells differ from cycling G2 cells by a higher total protein content. Cells with an elevated total protein content are found among 2c cells too, and these also fail to synthesize DNA in response to stimulation to proliferation. It is concluded that the quantity of protein accumulated in the cytoplasm may be one of the factors controlling cell proliferation.