[Poly(ADP-ribose) polymerase (PARP): physiological and pathological roles].

PARPs are a large family of 18 enzymes found in most eukaryotes. PARP-1, the most abundant isoform, is activated by DNA breaks and catalyzes the post-translational modification of proteins. It forms polymers ofADP-ribose and attaches them to acceptor proteins, including histones, DNA repair proteins, transcription factors. PARP-1 is a key enzyme involved in a maintenance of genomic stability. Excessive activation of the enzyme has been shown to contribute to tissue injury and inflammatory disorders. PARP is a key mediator of cell death in oxidative stress, ischemia and DNA damage. It also promotes the activation ofproinflammatory gene expression. Inhibition of PARP-1 provides significant protection in animal models of cardiovascular, autoimmune and inflammatory diseases. PARP inhibitors have shown antitumor activity because they compromise ability of cancer cells to repair DNA. PARP-1 is a promising therapeutic target.

[Effect of NF-kappaB activation inhibitor curcumin on the oogenesis and follicular cell death in immune ovarian failure in mice].

In experiments on CBA mice, we studied the influence of an inhibitor of nuclear transcription factor kappaB activation curcumin, obtained from Curcuma longa, on the meiotic maturation of oocytes and apoptotic and necrotic death of follicular cells at immune ovary failure induced by immunization of animals with allogenic ovarian extracts. NF-kappaB plays a pivotal role in the induction of genes encoding pro-inflammatory factors (cytokines, adhesion molecules, inducible NO-synthase and cyclooxygenase) and in regulation of cell proliferation and death. It has been shown that immunization of mice increased the death of follicular cells through anapoptotic and necrotic pathways, which led to inflammatory response (according to blood leukogram and impairment the oocyte meiotic maturation at metaphase I and II). Intragastric administration of curcumin (Sigma, USA, 2 mg of the mouse weight, four times a week during the period of immunization) reduced the number of the follicular cells died through apoptotic and especially necrotic pathway. Curcumin attenuated an inflammatory response and improved the meiotic maturation of oocytes impaired under experimental immune ovarian failure in mice.

[Functional activity of peritonal macrophages in liver immune damage of cellular and antibody genesis in mice].

The aim of present work was to compare the functional activity of peritoneal macrophages (Mf) at T-cellular and antibody induced hepatitis in mice of CBA line. T-cellular hepatitis was caused by concanavalin A (ConA), antibody-induced hepatitis was caused by administration of xenogenic anti-liver antibodies: gamma-globulin fractions of antihepatocytotoxic serum (gamma-AHCS). It was found that single injection of ConA or gamma-AHCS caused damage of liver with cytolytic syndrome through 20 hours. Functional activity of Mf in these conditions was significantly different. Application of ConA resulted in the decrease in phagocytosis of latex particles and oxygen-dependent metabolism; application of gamma-AHCS--to increase of these processes. Weakening of Mf activity may be one of the reasons for the decrease of dead cell eliminations that results in the maintenance of inflammatory reaction. At the same time significant amplification of phagocytic Mf activity may be one of the pathways of free radical endogenic sources increase that causes cell alteration and plays its role as mediators at inflammation.

[Proliferation and death of liver mononuclear cells in immune lesions induced by concanavalin A and anti-liver antibodies in mice].

Two types of experimental liver failure in mice were investigated to study the immune mechanisms of liver disease: 1) T-cell-mediated injury induced by administration of concanavalin A (ConA) and 2) antibody-mediated injury induced by administration of anti-liver antibodies (ALA, gamma-globulin fraction of sera from rabbits immunized with liver tissue). It was established, that both types of liver injury were accompanied by the activation of immune processes in the liver, as shown by the increase of liver mononuclear cell proliferation, estimated using IPO-38 monoclonal antibodies. In contrast to ConA treatment, the immune activation under ALA-treatment was also associated with the increase in the percentage of plasma cells and small lymphocytes in liver mononuclear cells. At the same time, an increase in apoptotic and necrotic mononuclear cell death was more pronounced under ConA-treatment. This was accompanied by enhanced Fas receptor expression in these cells. Thus, it was shown that in case of T-cell mediated liver injury, the balance between cell proliferation and cell death in mononuclear liver cells was shifted toward the significant increase of apoptotic and necrotic cell death, particularly Fas-mediated apoptosis, while immune processes activation and cell proliferation were more pronounced in the case of antibodies-mediated injury.

[The protective effect of molsidomin in immune ovarian pathology in mice].

Experimental immune ovarian failure in CBA mice was induced by either administration of xenogenic anti-ovarian antibodies (scheme 1) or immunization with allogenic ovarian extracts (scheme 2). It was shown that both types of treatment impaired the meiotic maturation of oocytes: the number of cells at the stages of metaphase I and metaphase II decreased compared to the cells of control mice. In both schemes of experiments, impaired oogenesis was accompanied by reduction of percentage of viable follicular cells and by increase in the part of cells possessing morphological features of apoptosis. In contrast, the number of necrotic follicular cells increased in scheme 2 only. The donor of nitric oxide molsidomin (10 mg/ kg), when injected an hour before administration of xenogenic anti-ovarian antibodies or allogenic ovary extracts, improved the meiotic maturation of oocytes and favored follicular, lymph nodes and thymus cells survival by decreasing the number of apoptotic and necrotic cells.

[Effect of thromboxane B2 on morphological features of apoptosis in cultured rat hepatocytes exposed to hepatotropic substances].

Thromboxanes (TX) are known to have damaging effect on a liver but their influence on the cell death, in particular on hepatocyte apoptosis and its morphological features is not investigated enough. Cell death of the rat hepatocytes was investigated in primary culture by double vital staining with fluorescent nuclear stains Hoechst 33342 and propidium iodide and by electron microscopy. It was established that exogenous Tx B2 increases the amount of hepatocytes with early stages of apoptosis - the condensed chromatin and nucleus and cell size reduction. The changes in a percentage of hepatocytes with morphological features of the late stages of apoptosis - fragmented nuclei and division on apoptotic bodies were not revealed. Tx B2 intensified the carbon tetrachloride action on hepatocyte apoptotic death and increased chromatin condensation. Tx B2 application to hepatocytes injured by chenodeoxycholic acid significantly increased the amount of cells with a final stage ofapoptosis.

[Death of the ovarian follicular cells in mice with impaired oogenesis of the immune nature].

An impairment of the meiotic maturation of the oocytes has been shown in vitro for 2 types of immune damage of the ovaries in mice induced by xenogenic antiovarial antibodies and immunization with allogenic ovaria. Impairment of the oogenesis was followed by the follicular cell death, primarily by on the apoptic way, but under the immunization with allogenic ovary a necrotic way of their death was also activated.

[Effect of high-dispersed silica on the cultured rat hepatocytes].

Highdispersed silica (HDS) is an active substance of medicine "Siliks", which demonstrates high protein adsorption property and is used for hatching out toxic agents of protein origin. To discover new potential applications of this substance, it is necessary to study the direct effect of HDS on cell viability. We studied the effects of HDS in concentration 0,0001%, 0,001% and 0,01% on cultured rat hepatocytes at 4 and 24 hours. To estimate the number of alive vs. apoptotic and necrotic cells, fluorescent nucleic dyes Hoechst 33342 and propidium iodide were employed. Cells that underwent autophagic processes were estimated using a specific fluorescent autophagosome marker monodansylcadaverine. We show here that HDS has dose-dependent effect on cell death; the number of cells with apoptotic features increased after 4 hours and in greater extend after 24 hours of treatment with HDS. At the lowest concentration HSD did not significantly affect cell viability, and we observed decrease in postapoptotic necrosis in cell culture. The highest concentration of HSD dramatically increased cell death through both necrosis and apoptosis. At the same time, autophagic activity was suppressed by 0,01% HSD.

[The role of nitric oxide in the development of humoral immune response in mice].

The immune response in CBA mice was evoked by injection of sheep erythrocytes. The number of antibody-producing cells in the spleen, as well as nitric oxide production, oxygen-dependent metabolism and 5"-nucleotidase activity of peritoneal macrophages and lymphocytes were studied on days 1-5-14 after immunization. It was shown that during the inductive phase of the immune response (day 1), the peritoneal cells increased nitric oxide production, while later their functional activity increased and NO level became normal. The use of NO-synthase inhibitors (non-selective L-NNA and iNOS inhibitors SMT and dexamethasone) increased the immune response and decreased the macrophage functional activity. The use of NO-donator SNP resulted in reverse effect: decrease of the immune response and stimulation of peritoneal cells functional activity. The data obtained indicate that nitric oxide participates in the immune response regulation, in particular, through the suppressive effect of macrophages.

[Effect of inhibitors of cyclooxygenase and lypoxygenase pathway of metabolism of arachidonic acid on the development and suppression of the immune response in mice]. / Vplyv blokatoriv tsyklo- ta lipoksyhenaznoho shliakhiv metabolizmu arakhidonovoï kysloty na rozvytok i supresiiu imunnoï vidpovidi u myshei.

The influence of indomethacin (IM) and nordihydroguaiaretic acid (NDGA) as inhibitors of cyclooxygenase and lypoxygenase pathways of arachidonic acid metabolism, accordingly, on the development of the immune response (IR) to sheep red blood cells (SRBC), as well as on the formation and functional activity of the antigen-induced (by the tolerogenous dose of SRBC) T-suppressors (AITs) was studied. Investigation was carried out on mice of line CBA. The IR was estimated by quantity of antibody-forming cells in the mouse spleen. The functional activity of AITs was determined in the transfer adopting system by the intensity of IR suppression in mice--recipients. The data obtained have demonstrated, that both inhibitors mainly stimulated the IR which was more expressed at NDGA application and depended on a phase of the IR. The stimulation of the IR was related with a suppression of AITs and a decrease in their functional activity.

[Lipoxygenase inhibitors induce rat hepatocyte apoptosis in primary culture]. / Blokatory lipoksyhenaz indukuiut' apoptoz hepatotsytiv shchuriv u pervynnii kul'turi.

Arachidonic acid metabolites have been shown to have a wide range of effects on cell proliferation and viability. In this study, the effects of lipoxygenase (LO) inhibitors nordihydroguaiaretic acid (NDGA) and caffeic acid (CA) on the viability of cultured rat hepatocytes (HC) were investigated. As a result, treatment with NDGA and CA for 4 h and 24 h decreased ALT release from HC and increased a number of apoptotic cells. Apoptosis inducing effects of general LO inhibitor NDGA were more pronounced, than those of 5-LO inhibitor CA. The results suggest that lipoxygenase pathway of arachidonic acid metabolism, in particular 5-LO, is essential regulator of hepatocyte survival and apoptosis.

[Effect of exogenous leukotrienes and lipoxygenase inhibitors on apoptosis and necrosis in cultured rat hepatocytes]. / Vplyv ekzohennykh leikotryieniv i blokatoriv lipoksyhenaz na apoptoz ta nekroz hepatotsytiv shchuriv u pervynnii kyl'turi.

Liver cell death by apoptosis and necrosis occurs upon the liver injury. Lipoxygenase pathway of arachidonic acid metabolism is known to regulate the viability and apoptosis in some cell types, but its role in hepatocyte cell death is not fully understood. We studied the influence of leukotrienes (LT) and lipoxygenase inhibitors on apoptosis and necrosis in rat hepatocyte primary culture by double staining with Hoechst 33342 and propidium iodide and electron microscopy. Treatment with general lipoxygenase inhibitor nordihydoguaiaretic acid and 5-lipoxygenase inhibitor caffeic acid (2. 10(-5) M) for 4 and 24 h induced hepatocyte apoptosis. LTB4 and LTC4 (10(-8) M) decreased the number of living cells and increased the number of necrotic cells. LTs exerted the same necrotic effect on hepatocytes, treated with lipoxygenase inhibitors. It is important that LTs decreased apoptosis induced by inhibitors treatment. These data suggest that lipoxygenase pathway of arachidonic acid metabolism is important regulator of hepatocytes viability and apoptosis The increase of lipoxygenase product formation, in particular LTs, may diminish apoptosis and increase necrosis in hepatocytes upon the liver injury.

[From Bogomoletz anti-reticular cytotoxic serum to monoclonal antibodies]. / Vid antyretykuliarnoï tsytotoksychnoï cyrovatky Bohmol'tsia do monoklonal'nykh antytil.

The paper devoted to the 120th anniversary of A.A. Bogomoletz birth deals with the developmental stages of the cytotoxin (xenogenic antibody) doctrine from creating the anti-reticular cytotoxic serum (ACS) of Bogomoletz to the use of monoclonal antibodies both in biological investigations and in medicine. The use of ACS for treating various diseases and the mechanisms of its effect; obtaining other anti-organ and anti-tissue cytotoxic sera and establishing their experimental effects; the use of a set of cytotoxic sera, taken in both inhibitory and stimulating doses, in medicine and agriculture, as well as the drawbacks of cytotoxic sere (polyclonal xenogenic antibodies) are described. The experience of using monoclonal antibodies to identify the structures of protein antigenic determinants and to establish their role in different diseases are analysed. The use of monoclonal antibodies for treating various diseases is reviewed. The advances in the techniques of obtaining monoclonal antibodies are noted.

[Effect of inhibitors of cyclooxygenase and lipoxygenase pathways of arachidonic acid oxidation on the immune response and the activity of the monoxygenase system and lipid peroxidation in the spleen and liver in mice]. / Vplyv blokatoriv tsyklooksygenaznogo ta lipoksygenaznogo shliakhiv metabolizmu arakhidonovoï kysloty na imunnu vidpovid', aktyvnist' monooksygenaznoï systemy i perekysnogo okysnennia lipidiv u selezintsi ta pechintsi myshei.

In experiments on CBA mice we studied an immune response (IR) to sheep red blood cells, the activity of monooxygenase system and lipid peroxidation (LP) in a spleen and a liver after administration of indomethacin (IND) and nordihydroguaiaretic acid (NDGA) as the inhibitors of cyclooxygenase and lypoxygenase pathways of oxydation of arachidonic acid consequently. We have found that the both inhibitors changed differently the intensity of IR during its development. IND and NDGA activate the accumulation of antibody-forming cells in the mouse spleen in a dose-dependent fashion at both the inductive and fading phases of IR. At the productive phase these changes are less expressed and they are different depending on the dose of NDGA: the smaller dose increases the immune response and the bigger one decreases it a bit. Changes in the activity of the monooxygenase system in spleen and liver, affected with the both inhibitors, independing on the dose, were of different direction: after IND administration the activity increased, but after NDGA administration it decreased at all the terms of investigation, excluding the term of the 5-th day (productive phase of IR). In these conditions changes in the activity of IR were of opposite direction as compared to the changes in the monooxygenase system.

[The in-vitro resumption of meiotic maturation and of the first polar body by mouse oocytes at different stages of the estrous cycle]. / Vidnovlennia in vitro meiotychnoho dozrivannia ta formuvannia pershoho poliarnoho til'tsia ootsytamy myshei na riznykh stadiakh estral'noho tsyklu.

Resumption of the meiotic maturation and extruded the first polar body of the murine oocytes was registered at 2 and 14 hours under culturing in the medium containing 1.7 mM Ca2+. There were differences between the groups in the percentage of GV oocytes with zona pellucida that acquired competence to undergo germinal vesicle breakdown (GVBD) at 2 hr time point 22.2% of "small" and 32.4% of "large" follicles of mouse at dioestrus and, respectively, 28.1% and 41.8% follicles of mouse at oestrous underwent GVBD. At 14 time point 21.4% of "small" and 30.3% of "large" follicles of mice at mice at dioestrus and, respectively, 26.1% and 43.6% follicles of mouse at oestrous underwent GVBD and (then) extruded the first polar body (FPB). It may be suggested, that acquisition of competence of the oocytes to resume meiosis in vitro depends on the size of follicles and the stage of murine oestrous cycle. Thus, the largest percentage of GV oocytes resuming meiosis was from "largest" follicles of mouse at oestrous.

[Effect of cultured mice hepatocytes on the proliferation of immunocompetent cells]. / Vplyv faktoriv, shcho vydiliaiutsia kultyvovanymy hepatotsytamy myshei, na proliferatsiiu imunokompetentnykh klityn.

Mice hepatocytes in primary cultures were treated during 2 h with normal rabbit antibodies (0.5 mg/ml), antiliver antibodies (0.5 mg/ml) and CCl4 (5 mM). Functional state of cells was studied according to DNA, RNA and protein synthesis. Cell-free supernatants were collected during 24 h after cells treatment. Spontaneous proliferation in vitro of syngeneic cells from the spleen, thymus, lymph node and bone marrow cultured with hepatocyte supernatants was investigated. Cultured intact hepatocytes were shown to produce factors that changed proliferation of the thymus and spleen cells. Hepatocyte supernatants demonstrated different effect on cells from different immune organs. Hepatocyte function alteration (minimal under normal antibodies treatment and maximal under CCl4 treatment) changed liver cells humoral influence on the immune cells.

[Changes of neutrophil functions resulting from intact and injured liver cells in mice]. / Zminy funktsionalnoi aktyvnosti neitrofiliv pid vplyvom faktoriv, shcho vydiliaiutsia intaktnymy ta urazhenymy klitynamy pechinky myshei.

Separately cultured parenchymatous and nonparenchymatous liver cells of mice produce factors to influence migration properties and oxygen-dependent metabolism of neutrophils. Cell-free supernatant of intact parenchymatous cells activates neutrophil chemotaxis only, while supernatant of intact nonparenchymatous cells stimulates oxygen-dependent reactions as well. Injure of the liver cells with 5 mM tetrachlormethane significantly increases neutrophil functional activation. Treatment of the liver cells with anti-liver antibodies (0.5 mg/ml) results in suppression of neutrophil activation. At the same time, supernatant of the liver cells treated with normal rabbit antibodies (0.5 mg/ml) possessed neutrophil stimulating properties, nonparenchymatous cell factors stimulating mainly the neutrophil oxygen-dependent metabolism.