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Volume 11, Nº 4 (2017)

Article

Apoptosis and autophagy in hepatocarcinoma cells induced by different forms of lithium salts

Bgatova N., Gavrilova Y., Lykov A., Solovieva A., Makarova V., Borodin Y., Konenkov V.

Resumo

Hepatocellular carcinoma (HCC) is an aggressive cancer that is resistant to drug therapy. It is believed that the development of HCC is correlated with misregulation of programmed cell death. Discovery of effective inducers of HCC cell death is very important for HCC therapy. The aim of this work was to identify structural changes leading to the death of HCC cells exposed to nanosized and original forms of lithium salts. Structural features of autophagy and apoptosis were revealed in HCC cells after their incubation with various forms of lithium salts by light, electron microscopy, and flow cytometry. It was shown that nanosized forms of lithium carbonate and lithium citrate had a pronounced effect on HCC-29 cells. Of these forms, the nanosized lithium citrate induced mainly apoptosis, while the nanosized form of lithium carbonate, along with apoptosis, induced autophagic death of HCC cells.

Cell and Tissue Biology. 2017;11(4):261-267
pages 261-267 views

The MEK/ERK pathway is essential for maintenance of cytoprotective autophagy in E1A+cHA-RAS transformants after exposure to radiation

Kochetkova E., Blinova G., Zubova S., Bykova T., Pospelov V., Pospelova T.

Resumo

Autophagy is a conserved process of protein and organelle degradation that serves to maintain cell viability. Autophagy is frequently induced in response to stress or to exposure to DNA-damaging agents or retinoids, as well as to starvation and deficiency of growth factors. In this work, autophagy induced in E1A+cHA-RAS transformed cells in response to X-ray radiation was studied, with a focus on the role of the MEK/ERK signaling pathway in the regulation of radiation-induced autophagy. It was found that inhibition of the MEK/ERK pathway diminished cell viability and altered the sequence of events in radiation-induced autophagy. In particular, it caused aberrations in its final stages, leading to cytoplasmic accumulation of the p62/SQSTM1 adaptor protein in autophagic cavities of unclear origin. Thus, the MEK/ERK pathway activity is essential for the induction and maintenance of autophagy, increasing the viability of exposed cells in response to radiation.

Cell and Tissue Biology. 2017;11(4):268-274
pages 268-274 views

Structural modifications of macrophages initiated by tick-borne encephalitis virus

Drobot E., Lyapun I., Plekhova N., Pustovalov E., Somova L., Leonova G.

Resumo

Macrophages are cells of natural immunity and play a key role in pathogenesis of viral infections. Results of ultrastructural research on macrophages infected with tick-borne encephalitis virus (TBEV), an agent that causes dangerous infections affecting nervous system in human beings and belongs to the Flaviviridae family, were shown here. Using virology methods, it was ascertained that the TBEV is consumed by macrophages and multiplies in them. Ultrastructural research showed that the virus penetrates into the cytoplasm by means of local plasmalemma lysis and newly synthesized virus particles escape from the cell by the same path. At the same time, induration of the perinuclear space of cytoplasm was observed, where ribosomes, microfilaments, ribonucleoprotein threats, and virus-specific structures, namely, nucleocapsids, tube formations, and viroplasts, were found in large quantities. On the surface of viroplasts, newly synthesized virus particles were visualized. Thus, evidence was presented that microphages can play certain role in spreading of TBEV and are the target of the virus. Like active antigen-presenting cells, such macrophages can modulate the protective response of an organism and affect the pathogenesis of tick-borne encephalitis.

Cell and Tissue Biology. 2017;11(4):275-285
pages 275-285 views

The phenomenon of atypical DNA comets

Zhanataev A., Anisina E., Chayka Z., Miroshkina I., Durnev A.

Resumo

The phenomenon of atypical DNA comets observed in experiments using DNA-comet assay is described and illustrated. The current hypotheses explaining the nature of atypical DNA comets and our own take on the issue are considered. The practical importance of the registration of atypical DNA comets during the assessment of genotoxicity is discussed.

Cell and Tissue Biology. 2017;11(4):286-292
pages 286-292 views

Heterogeneity of coilin-containing nuclear domains in early mouse embryos

Bogolyubova I.

Resumo

The nucleus of two-cell mouse embryos contains coilin-containing bodies of two types: (i) one to three large spherical structures 1 μm in size and (ii) small foci, which vary in number in different blastomeres. The largest coilin-containing structures, unlike the smallest ones, contain RNA polymerase I, nucleic acid chaperon YB-1, and actin. Neither large nor small coilin-positive domains contain symplekin, one of the signature components of histone locus bodies. In nuclei of late two-cell embryos, symplekin localizes to one to two well-formed spherical bodies that are observed both in close proximity to the coilin-positive structures and apart from them. Large coilin-containing bodies were not observed in embryos at the morula stage, as well as in the nuclei of late two-cell embryos after artificial suppression of transcription activity. Thus, the population of coilin-containing bodies in the nuclei of late two-cell embryos of mice is heterogeneous in morphology and molecular composition. It can be suggested that the largest coilin-containing bodies are provisional nuclear domains that are formed against a background of significant changes in the nuclear metabolism at the final stages of embryonic genome activation and the initial stages of reactivation of nucleolar transcription.

Cell and Tissue Biology. 2017;11(4):293-299
pages 293-299 views

An immunohistochemical study of depigmented skin of vitiligo patients

Revishchin A., Panteleev D., Zaharova L., Lomonosov K., Pavlova G.

Resumo

Immunohistochemical analysis was used to study depigmented areas, macular depigmentation, and perimacular areas of the skin of vitiligo patients. It has been shown that cells containing the melanocytic cell marker TRP1 are located in both macular and perimacular areas. Within the macula of depigmentation, all TRP1-positive cells are in close contact with the base membrane. In perimacular areas, many TRP1-positive cells that have lost contact with the base membrane are located in deep layers of epidermis. About 92% of TRP1-positive perimacular cells were also vimentin-positive. Vimentin-positive cells were numerous in perimacular areas, but missing in the macula of depigmentation. Dense groups of cells that were immunopositive for the transcription factor Snail, an inducer of epithelial mesenchymal transition, were localized in perimacular areas in close proximity to the macula of the depigmentation border. Within the macula of depigmentation, such cells were extremely rare. It seems reasonable that an intense process similar to the epithelial mesenchymal transition may be the cause of melanocyte death in the perimacular area and, thus, prevents repigmentation of depigmented areas.

Cell and Tissue Biology. 2017;11(4):300-307
pages 300-307 views

Karyotyping of Amoeba proteus

Demin S., Berdieva M., Podlipaeva Y., Yudin A., Goodkov A.

Resumo

In this paper, we have developed a method to prepare spreads of mitotic chromosomes of Amoeba proteus and described the process of Amoeba proteus karyotyping. This protocol allows obtaining spread chromosomes with a characteristic pattern of chromomeres on individual chromosomes. It is shown that, in the metaphase of mitosis, amoebas of strain B (one of the type strains of A. proteus in the Amoebae Cultures Collection of the Institute of Cytology, Russian Academy of Sciences) contain 27 pairs of chromosomes. It is established that the pattern of chromomeres is a chromosome-specific feature. A typical karyogram and image bank of DAPI- and YoYo1-banded individual chromosomes of A. proteus, strain B composed of five different spreads of mitotic cells are presented.

Cell and Tissue Biology. 2017;11(4):308-313
pages 308-313 views

Features of the embryonic development of Dienia ophrydis (Orchidaceae)

Kolomeitseva G., Ryabchenko A., Babosha A.

Resumo

A new Dienia type of the embryogenesis of orchid plants differing from the Liparis type, earlier observed for the tribe Malaxideae, has been described in Dienia ophrydis (J. Köenig) Seidenf. (Orchidaceae). The Dienia-type embryogenesis is characterized by the following features: (1) development of a single-celled suspensor formed by a cb-derivative, (2) linear arrangement of embryo cells at the tetrad stage, (3) atypical origin of some tiers, and (4) no divisions of the ci and cb cells. A hypothesis about the convergent similarity between the Dienia and Caryophyllaceae types of embryogenesis has been proposed. A number of embryo sac and embryo structures typical for D. ophrydis, including “petassum,” “fitting,” and “suspensor mantle,” have been first described. A “petassum” represents the remains of cell walls of the pollen tube and probably the filamentous apparatus of synergids sealing the micropyle side of a fertilized embryo sac. The sole suspensor cell has a special appendix (“fitting”), which connects it to the embryo. The suspensor and the fitting are surrounded by a special envelope (“suspensor mantle”), which does not cover the basal cell of the embryo (ci).

Cell and Tissue Biology. 2017;11(4):314-323
pages 314-323 views

Reorganization of interphase microtubules in root cells of Medicago sativa L. during acclimation to osmotic and salt stress

Lazareva E., Baranova E., Smirnova E.

Resumo

We examined the organization of microtubule system of interphase cells in roots of Medicago sativa L. during acclimation to salt and osmotic stress at different concentrations of NaCl, Na2SO4, and mannitol. We identified morphological changes of tubulin cytoskeleton in different root tissues during the acclimation to salt and osmotic stress: (1) decreased density of the cortical microtubule network, (2) random orientation of cortical microtubule bundles, (4) thickening of the bundles, (3) nonuniform density of the bundles, (4) fragmentation of the bundles, and (5) formation of microtubule converging centers. Network thinning and thickening of the bundles were observed both under osmotic and salt stress. Random orientation of cortical microtubules was visualized under osmotic stress but not during salt stress. Fragmentation of microtubule bundles took place under salt stress with a high concentration of mannitol. Formation of microtubule converging centers was common under prolonged action of sodium sulfate, less evident under sodium chloride, and not found after mannitol treatment. Our data show that, in alfalfa root cells, cortical microtubules rearrange not only in response to different ions, but also to osmotic pressure. Thus, the signaling pathways and molecular mechanisms inducing reorganization of the microtubule system may be triggered by sodium cations, as well as by sulfate and chloride anions at concentrations that do not cause irreversible cell damage.

Cell and Tissue Biology. 2017;11(4):324-334
pages 324-334 views

The dipole-modifying effect of styrylpyridinium dyes and flavonoids on model membranes of different lipid compositions

Efimova S., Schagina L., Ostroumova O.

Resumo

Changes in dipole potential of lipid bilayers ϕd mimicking cell membranes induced by the adsorption of low-molecular-weight amphiphiles, flavonoids (phloretin and quercetin), and styrylpyridinium dyes (RH 421 and RH 237) were measured. A method based on the determination of ionophore-induced transmembrane current was used to evaluate changes in ϕd after modifier addition. The characteristic parameters of the Langmuir adsorption isotherm and the greatest changes in ϕd at an infinitely large concentration of flavonoid and its desorption constant, which reflects the affinity of the flavonoid to the lipid phase, were determined. The slopes of linear dependences of ϕd increasing on the concentration of the styrylpiridinium dyes in membrane-bathing solution were defined. It was found that the dipole-modifying effect of phloretin depends on the charge of the lipids forming the membranes, while the ability of quercetin to reduce ϕd is determined by the initial hydration of the bilayer. The results indicate that there are different mechanisms of the decrease in ϕd upon the adsorption of the tested flavonoids. It was shown that the changes in ϕd at the incorporation of styrylpyridinium dyes into bilayers are determined by the interaction of modifiers with membrane components.

Cell and Tissue Biology. 2017;11(4):335-341
pages 335-341 views

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