RESUMO
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.
Assuntos
Nucléolo Celular , Hepatócitos , Fígado , Radiação Ionizante , Alanina Transaminase/metabolismo , Animais , Aspartato Aminotransferases/metabolismo , Nucléolo Celular/enzimologia , Nucléolo Celular/efeitos da radiação , Creatina Quinase/metabolismo , Hepatócitos/enzimologia , Hepatócitos/efeitos da radiação , Fígado/enzimologia , Fígado/efeitos da radiação , Masculino , Ploidias , Purina-Núcleosídeo Fosforilase/metabolismo , Ratos , Irradiação Corporal TotalRESUMO
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.
Assuntos
Compostos de Alumínio/farmacologia , Antineoplásicos/farmacologia , Cloretos/farmacologia , Neuroblastoma , Células Neuroepiteliais/efeitos dos fármacos , Neurotoxinas/farmacologia , Tretinoína/farmacologia , Adaptação Fisiológica , Cloreto de Alumínio , Animais , Astrócitos/citologia , Astrócitos/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Divisão Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , DNA/análise , Teoria da Informação , Células Neuroepiteliais/patologia , RNA/análise , Células Tumorais CultivadasRESUMO
The aim of the study was to investigate serum levels, as well as the ability to produce in vitro, some pro-inflammatory and anti-inflammatory cytokines in patients with brain tumours (BT). Serum concentrations of IFN-gamma, IFN-alpha, TNF-alpha, IL-4, IL-6, IL-8, and RA IL-1 were determined in 17 patients with brain tumours, and in 21 healthy donors. The results showed that baseline levels of cytokines studied was above normal in most patients (78%) had the highest rates of IFN-gamma. During the whole observation period, slightly higher in patients compared with healthy individuals (t=1.8) was the content of IFN-alpha. Serum concentrations of IL-4 in patients before treatment were significantly higher than in control group (p<0.01), remaining at about the same level, and after treatment. Levels of Rail-1, IL-6 and IL-8 in patients before treatment were within normal limits, while remaining unchanged at the end of the survey. Indicators induced in vitro production of cytokines studied to some extent correlated with their serum levels. Hyperproduction in vitro IFN-gamma was observed in 61% of patients, low production - 28% of patients, and only in 11% of cases, these figures were within normal limits. In patients with malignant brain tumors were identified violations of serum levels and production of cytokines studied. The most significant was the increase of PHA-induced production of IFN-gamma, which was not reduced by the end of treatment. High serum and, in particular, PHA-induced levels of IFN-gamma at the end of treatment may be recommended as an additional criterion for immunodiagnosis in a dynamic laboratory monitoring.
Assuntos
Neoplasias Encefálicas/sangue , Neoplasias Encefálicas/diagnóstico , Citocinas/sangue , Adulto , Citocinas/metabolismo , Feminino , Humanos , Interferon-alfa/sangue , Interferon-alfa/metabolismo , Interleucina-4/sangue , Interleucina-4/metabolismo , Interleucina-6/sangue , Interleucina-6/metabolismo , Interleucina-8/sangue , Interleucina-8/metabolismo , Masculino , Pessoa de Meia-Idade , Receptores de Interferon/sangue , Receptores de Interferon/metabolismo , Fator de Necrose Tumoral alfa/sangue , Fator de Necrose Tumoral alfa/metabolismo , Receptor de Interferon gamaRESUMO
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).
Assuntos
Febre Suína Africana/patologia , Linfócitos/patologia , Tecido Linfoide/patologia , Monócitos/patologia , Febre Suína Africana/imunologia , Febre Suína Africana/virologia , Vírus da Febre Suína Africana/fisiologia , Animais , Apoptose , Medula Óssea/patologia , Técnicas de Cultura de Células , Núcleo Celular/patologia , Forma Celular , DNA/análise , Linfócitos/imunologia , Tecido Linfoide/imunologia , Monócitos/imunologia , Suínos , Tetraploidia , Fatores de Tempo , TriploidiaRESUMO
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.
Assuntos
Actiemil/farmacologia , Divisão Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Nucléolo Celular/efeitos dos fármacos , Núcleo Celular/efeitos dos fármacos , Meios de Cultura , Citoplasma/efeitos dos fármacos , Células HeLa , Humanos , Fatores de TempoRESUMO
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.
Assuntos
Diferenciação Celular , Linhagem Celular Tumoral/citologia , Morte Celular , Linhagem Celular Transformada , Nucléolo Celular/genética , Nucléolo Celular/metabolismo , Nucléolo Celular/ultraestrutura , Núcleo Celular/genética , Núcleo Celular/metabolismo , Núcleo Celular/ultraestrutura , Análise por Conglomerados , DNA/metabolismo , Humanos , Citometria por Imagem , Índice Mitótico , PloidiasRESUMO
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.
Assuntos
Células CACO-2/citologia , Contagem de Células , Técnicas de Cultura de Células , Divisão Celular , Nucléolo Celular/genética , Núcleo Celular/genética , Humanos , Citometria por Imagem , Poliploidia , Fatores de TempoRESUMO
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.