Autophagy in Hepatocytes during Distant Tumor Growth.

Structural changes in the liver of CBA mice were studied during the development of experimental hepatocarcinoma-29 inoculated into the hip. A decrease in the volume density of hepatocyte cytoplasm, mitochondria, endoplasmic reticulum, and lipid inclusions and an increase in the volume density of lysosomal structures during tumor growth were observed. All stages of intracellular autophagy were recorded by the method of electron microscopy. These stages included the appearance of autophagosomes, autophagolysosomes, and secondary lysosomes in the hepatocyte cytoplasm. Fragments of cytoplasm, glycogen rosettes, mitochondria, and fragments of endoplasmic reticulum with ribosomes were found in autophagosomes. The obtained data indicate the development of non-selective autophagy in the liver during distant tumor growth in aimed at the maintenance of intracellular homeostasis in hepatocytes and energy and trophic homeostasis of organism.

Effects of Bone Marrow Multipotent Mesenchymal Stromal Cells and Their Secretory Products on Microcirculation in the Broad Ligament of the Uterus of Wistar Rats during Experimental Chronic Genital Inflammation.

Effects of bone marrow multipotent mesenchymal stromal cells and their secretory products released into the conditioned medium on microcirculatory bed in the broad ligament of the uterus were studied in Wistar rats with chronic genital inflammation. Opposite changes in the parameters of microcirculation and lymphatic drainage in the broad ligament of the uterus were observed after administration of cells and conditioned medium via different routes, which should be taken into account during the treatment of inflammatory and degenerative processes in the pelvic organs.

APOPTOSIS AND AUTOPHAGY IN HEPATOCARCINOMA CELLS INDUCED BY DIFFERENT FORMS OF LITHIUM SALTS.

Hepatocellular carcinoma (HCC) is one of the aggressive and resistant to drug therapy cancer. It is believed that the development of HCC is correlated with dysregulation of programmed cell death. So, the search for effective inducers of HCC cell death is very important. The aim of the work was to identify structural changes leading to HCC cell death when exposed to nanoscale and original forms of lithium salts. Using light and electron microscopy and flow cytometry, structural features of autophagy and apoptosis in HCC cells after their incubation with various forms of lithium salts has been revealed. It is shown that a more pronounced effect on HCC-29 cells have nanoscale forms of lithium carbonate and lithium citrate. At the same time, nanoscale lithium citrate mainly induced apoptosis, and nanosized form of lithium carbonate, along with apoptosis, induced autophagic death of HCC-29cells.

Expression of Molecular Markers of Angiogenesis, Lymphangiogenesis, and Proliferation Depending on the Stage of Skin Melanoma.

The expression of molecular markers characterizing activity of the tumor process and metastases (proliferation marker Ki-67, angiogenesis marker CD34, and lymphangiogenesis markers podoplanin and LYVE-1) was assessed by immunohictochemical method in the primary tumor specimens collected during surgery for cutaneous melanoma (40 patients). Proliferative activity of the tumor tissue and volume density of peritumoral blood and lymph vessels increased with increasing tumor malignancy, which could indicate the risk of metastases.

Cell Composition of Central and Peripheral Lymphoid Organs of Wistar Rats Treated with a Biomedical Cell Product.

We studied the effect of a biomedical cell product (bone marrow multipotent mesenchymal stromal cells and products secreted by these cells in conditioned medium) on subpopulation composition of lymphoid cells of central and peripheral lymphoid organs of Wistar rats under normal conditions. Changes in the subpopulation composition of lymphoid organs depended on the route of administration of biomedical cell product.

[PERIPHERAL LYMPHOID STRUCTURES: FORMATION AND FUNCTION].

The problem of the formation of new lymphoid structures (neolymphogenesis) is quite controversial and widely discussed in the literature. Under normal conditions, lymphoid organs arise only in the process of fetal development (organogenesis), however in long-standing chronic inflammatory processes, nonhealing wounds, autoimmune diseases, oncologic pathology spontaneous formation of new lymphoid structures was noted. The structures of the peripheral lymphoid formations include the lymphocytes arranged singly and in clusters (infiltration), lymphoid nodules and lymph nodes. The morphogenesis of the components of lymphoid tissue and the possibility of creating artificial lymphoid structures, reproducing the function of the natural ones, is demonstrated. Important role in the development of lymphoid structures is played by mediators of inflammation, cytokines of the family of lymphotoxins, tumor necrosis factor. The possibilities of prosthetic substitution of the functions of the lymphoid structures are described for the activation of protective processes in the body.

Morphological Criteria of Cell Differentiation Stages in Experimental Hepatocarcinoma and Evaluation of Antitumor Drug Efficiency.

Structural polymorphism of 5 cell differentiation stages of hepatocarcinoma-29 from ascitic fluid is detected and the morphological criteria for identification of these stages are defined on the base of optic and electron microscopy findings, cytofluorometry, and DNA cytometry. The percentage of cells at differentiation stages 4 and 5 in the tumor structure increases after hepatocarcinoma cell inoculation into the hip. Injection of a cell cycle-modulating substance to animals with tumor growth shifts the proportion of cells with various differentiation stages. The morphological criteria of 5 stages of hepatocarcinoma-29 cell differentiation can be used for prospective drug testing.

[Effects of lithium carbonate nanosized particles on oxidant-antioxidant status in tumor tissue of hepatocarcinoma-29].

Oxidant-antioxidant status in tumor tissue of male-mice CBA at spontaneous course of hepatocarcinoma-29 and after repeated injections of lithium carbonate nanosized particles was evaluated on changes of lipid peroxidation (LPO) products level reacted with 2-thiobarbituric acid (TBA) as indicator of oxidative stress and activity of superoxide dismutase (SOD) and catalase enzymes as indicators of antioxidant defense by spectrophotometer <> (Bio-Rad, USA). Tumor development after hepatocarcinoma-29 cells injection into muscle right leg changed the levels of LPO activity in two-phase manner. TBA-active products content were decreased in 2,4 times in comparison with the control indicates after invasion of tumor cells, it was raised in 2,1 times at excessive tumor growth and diminished at terminal stage. Catalase activity was significantly elevated, but SOD activity was reduced in tumor tissue samples at active growth of hepatocarcinoma. The repeated injections of lithium carbonate nanosized particles at hepatocarcinoma inhibited processes of lipid peroxidation in tumor tissue in 2,4 times, but didn't influence on activities of catalase and SOD. Thus the effects lithium carbonate nanosized particles injections referred on maintenance of balance between the oxidant and antioxidants may be of some help to limit the progression of precancerous condition toward malignancy and tumor growth.

Parameters of Microcirculation in the Broad Ligament of the Uterus in Wistar Rats after Injection of Autologous Biomedical Cell Product.

We studied the effects of autologous biomedical cell product (bone marrow multipotent mesenchymal stromal cells and their conditioned media) on the parameters of the microcirculatory bed in the broad ligament of the uterus of normal Wistar rats were studied. The parameters of microcirculation and lymph drainage in the broad ligament changed in opposite directions in response to injection of autologous biomedical cell product via different routes. This fact should be taken into consideration when prescribing cell therapy for inflammatory degenerative processes in the pelvic organs.

Effects of Nanosized Lithium Carbonate Particles on the Functional Activity of Macrophages During Development of Hepatocarcinoma 29.

The functional activity of macrophages in response to injection of nanosized lithium carbonate particles after initiation of hepatocarcinoma 29 in male CBA mice was evaluated by the production of NO, arginase activity, and absorption of zymosan granules. In intact animals, NO production by peritoneal macrophages increased by 4 times and arginase activity 3.1 times in response to a single injection of nanosized particles into the hip muscle. The level of NO production by macrophages remained high after 4 and 5 injections, while arginase activity returned to normal. The level of phagocytic peritoneal macrophages increased by 1.4 times after 5 injections of the particles. The level of NO production by macrophages gradually increased in animals with hepatocarcinoma developing in the hip muscle: by 1.6 times on day 3, 3.2 times on day 7, and by 2.6 times on day 13 in comparison with the corresponding parameters in intact animals. The increase of NO production by peritoneal macrophages after tumor process initiation was not paralleled by changes in arginase activity and absorption of zymosan granules. The results indicated that injection of nanosized lithium carbonate particles after inoculation of hepatocarcinoma 29 cells in the right hip muscle tissue was inessential for the function of peritoneal macrophages by the studied parameters.

[Effects of lithium carbonate nanosized particles on nitric oxide production and arginase activity in tumor and peritoneal macrophages in hepatocellular carcinoma 29].

Three groups of male CBA mice were used. Group 1 consisted of intact mice. Hepatocarcinoma cells 29 (HCC-29) were transplanted into the right thigh muscle of animals group 2. Group 3 mice were injected 0.1 ml of lithium carbonate nanosized particles (NPs Li2CO3) at a dose of 0,058 mg on periphery of tumor growth one fold (3 day) and 5-fold (7 and 13 days). On day 7, numerical density of macrophages was raised in 5.8 times, the NO levels were increased by 1.9 times, and arginase activity was decreased in HCC tissue as compared with those values in normal liver. Tumor growth led to an increase in NO production by peritoneal macrophages (pMf) in 2.8 and 2.2-fold on day 7 and 13 days, respectively, and hadn't effect on arginase activity. A single injection of NPs Li2CO3 after inoculation tumor cells (3 days) didn't alter the NO levels rise in the tumor but five injections (7 days) increased it in 1.7 times as compared with values of mice group 2. The treatment of NPs Li2CO3 influenced the increasing the number density of macrophages in the tumor. Numerical density of macrophages in the tumor of mice group 3 was increased 9.6 and 1.6 times as compared with similar values in groups 1 and 2, respectively on 7 day after 5 injections of NPs Li2CO3. Treatment of NPs Li2CO3 after tumor cell transplantation didn't affect on the rise of NO levels and arginase activity in pMf. Thus, the effects of NPs Li2CO3, administered after HCC-29 cells transplantation, aimed at increasing activity of the NO-synthase way in HCC tumors and pMf, which can reduce the arginine levels required for tumor growth.

Thymus in experimental carcinogenesis of the prostate gland.

We studied structural changes in the prostate gland, thymus, and lymph nodes in CBA mice after transplantation of Ehrlich ascites tumor cells into the prostate gland. On experimental day 5, the number of blood and lymph vessels decreased in the gland; the percentage of connective tissue elements and glandular tissue and the number of immunoblasts in the thymus increased. On day 18, the number of blood vessels in the tumor decreased; the width of the cortex and glandular tissue increased in the thymus, while the number of immunoblasts was reduced. On day 28, tumor infiltration and increased number of lymphatic vessels in its stroma were observed; parenchyma was reduced, and the area of the connective tissue increased in the thymus. These structural changes indicated the development of accidental involution of the thymus during carcinogenesis of the prostate.

Effects of nanosized lithium carbonate particles on intact muscle tissue and tumor growth.

The effects of nanosized lithium carbonate particles on muscle tissue structure and development of experimental hepatocarcinoma-29 transplanted into the hip were studied in CBA mice. Necrotic changes in all structural components of the muscle were detected after intramuscular injection of nanosized lithium carbonate particles to intact animals. Regeneration of the muscle fibers after lithium carbonate treatment was associated with a significant increase in macrophage count, number of microvessels, activation of fibroblasts, and complete recovery of the organ structure. Injection of lithium carbonate nanoparticles at the periphery of tumor growth caused tumor cell necrosis, destruction of the vascular bed, and attraction of neutrophils and macrophages to the tumor focus. After the preparation was discontinued, the tumor developed with lesser number of vessels, smaller tumor cells, and lesser deformation of the cell nuclei structure.

Expression of intracellular molecular apoptosis regulator Bcl-2 in the liver in isolated and combined exposure to 24-h illumination and industrial frequency magnetic field.

We studied the effect of isolated and combined exposure to 24-h illumination and industrial frequency magnetic field on the expression of intracellular molecular apoptosis regulator Bcl-2 in the liver. Previous experiments showed enhanced expression of Bcl-2 protein in liver cells in animals maintained under conditions of 24-h illumination, whereas exposure to industrial frequency magnetic field had practically no effect on this process. Under conditions of combined exposure to 24-h illumination and industrial frequency magnetic field, magnetic field partially suppressed the expression of Bcl-2 induced by 24-h illumination.

Basal induction of cytochrome P-450 1A1/1A2 in cells of the liver, small intestinal villi, and lymph nodes in rats.

Rat liver, small intestinal wall, mesenteric, mediastinal, and popliteal lymph nodes of rats were studied by indirect immunoperoxidase method with monoclonal antibodies to cytochrome P-450 1A1/1A2. These cytochrome forms were detected in low concentrations in hepatocytes, macrophages of the small intestinal villi and mesenteric and mediastinal lymph nodes of intact animals. Basal induction of monooxygenase enzymes can be caused by cytochrome inductors entering with air (environmental pollution) or food.

The possibility of cytochrome P450 1A1/1A2 induction in cells of distant lymph nodes of rats after enteral treatment with benzo[a]pyrene.

The mesenteric, mediastinal, and popliteal lymph nodes of rats were studied by indirect immunoperoxidase method using monoclonal antibodies to cytochrome P450 1A1/1A2 after oral treatment with benzo[a]pyrene. These cytochrome forms were detected in monocytes, macrophages, reticular and littoral cells, cell detritus, and liquid contents of the paracortical and medullary sinuses of all studied lymph nodes. The results indicate that exo- and endogenous toxic substances are oxidized not only in the liver, but also in lymph nodes.