[Cytokine regulation in patients with brain tumours].

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.

[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.

[Immune reactivity and cytokine status in polytrauma].

The PHA-induced and serum levels of IFN-alpha and -gamma, TNF-alpha, IL-4, IL-6, IL-8, IL-RA, as well as CD3, CD4, CD8, CD20, CD56 lymphocytic subpopulations and serum IgG, IgM, and IgA were studied in 53 patients with multiple trauma to detect the most informative parameters of laboratory monitoring of the immunodeficiency developing during treatment. The stages of the study corresponded to the phases of development of an immune response: (1) an induction phase (days 1-4); (2) an immunoregulatory phase (days 5-10); (3) an effector phase (days 11-24). The findings suggest that at the early stage of the study, the drastic reduction was revealed in CD3, CD4 T-lymphocytic subpopulations, accompanied by inadequate humoral immunological parameters. By Stage 3 of the study, the majority of these parameters became normal; the serum levels of IgG and IgM remained lower only in patients with a fatal outcome. Throughout all the 3 stages of the study, the serum concentration of IFN-alpha remained in the almost normal ranges; there was no significant elevation in both the serum and induced levels of IL-4 either. The serum content of TNF-alpha in the examinees was significantly higher than the normal levels, by remaining virtually at the same level up to the end of treatment. The elevated serum levels of TNF-alpha were attended by its increased induction in vitro, without reducing up to Stage 3. From Stage 3, only some patients with a fatal outcome were observed to have the reduced induced and spontaneous levels of TNF-alpha. According to the findings, at Stage 1 of the study, the production of IFN-gamma in all patients was much increased, by significantly dropping by the end of treatment (by more than twice), without achieving, however, the normal levels, except the examinees with a fatal outcome, in whom the concentration of IFN-alpha increased or underwent no changes. A relationship was also found between the serum levels of IL-6 and IL-8 and the outcome of the disease: throughout the observation, their concentrations remained invariably high in patients with a fatal outcome whereas in those with a good outcome, there was a uniform decrease in the content of both cytokines by the end of the observation. Thus, the simultaneous increase in the levels of IFN-gamma, IL-6, IL-8, and TNF-alpha and accompanying reductions in CD3, CD4 and serum IgG, IgM are poor prognostic factors and may be used both as additional immunodiagnostic criteria and for immunotherapy monitoring in patients with polytrauma.

[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.

[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 experimental and clinical antitumor action of the interferon inducer larifan]. / Protivoopukholevoe deistvie induktora interferona larifana v éksperimente i klinike.

The paper presents the results of experimental and clinical studies on the antitumor effect of larifan. A statistically significant inhibition of growth of some continuous mouse tumors by larifan in a dose of 2.5 mg/kg given for 5-6 days and by a combination of larifan with an immunomodulator, inosiplex, tested on hepatoma XXIIa which has a rather low sensitivity to larifan. The antitumor effect of larifan may be due to interferon production and immunomodulating effect. Treatment with 0.05% larifan ointment of patients with cervical dysplasia of moderate degree resulted in a significant increase of previously diminished production of alpha-interferon after 10-14 vaginal tampons and clinical cure in 88% of cases. A course of 8-10-day larifan monotherapy given before radial therapy resulted in a considerable improvement in patients with cervical cancer of II-III stages, and subsequently, for 1 1/2-month combined radial therapy, no significant changes in production of alpha-IF were observed. At the same time, in patients with cervical cancer treated by radial therapy alone a significant decrease of alpha-IF production by blood mononuclears was observed by the end of therapy.