Galactose specific lectins from prostate tissue with different pathologies: biochemical and cellular studies.

BACKGROUND: Galectins-galactose-specific lectins are involved in various types of cell activities, including apoptosis, cell cycle regulation, inflammation and cell transformation. Galectins are implicated in prostate malignat transformation. It is not known yet if prostate glands with different grade of pathologies are expressing different galectins and if these galectins express different effects on the cell viability. METHODS: Cytosolic galactose-spesific lectin fractions from prostate tissue with different diagnosis were purified by affinity chromatography and analyzed by electrophoresis in polyacrylamide gel electrophoresis with sodium dodecyl sulphate. The lectin effects in a source-dependent maner were studied on cell viability on peripheral lymphocytes by MTT reduction method and on apoptosis by flow cytometry method. RESULTS: Affinity purified galactose-specific lectins fractions from normal and pathological tissue samples are characterized with different protein composition and they express different effects on cell viability and apoptosis. CONCLUSION: The effects of cytosolic galactose-specific lectins depend on the source of lectin fraction (glandular tissue disease). We suppose that the released cytosolic galectins from prostatic high grade intraepithelial neoplasia and adenocarcinoma tissue could suppress the immune status of the host patients.

Bio-Effects of TiO2 Nanoparticles on Human Colorectal Cancer and Umbilical Vein Endothelial Cell Lines

Background: Due to the possible biomedical potential of nanoparticles, titanium dioxide nanoparticles (TiO2 NPs) have received great attention in cancer research. Although selectivity of cytotoxicity with TiO2 NPs in various cells is clinically significant comparisons of cancer and non-cancer cells have been limited. Therefore, we here studied exposure to TiO2 NPs in colorectal cancer cells (CRCs) and human umbilical vein endothelial cells (HUVECs). Methods: After characterization of TiO2 NPs, culture and treatment of cells (HCT116, HT29 and HUVEC), viability was assessed by MTT assay and in terms of morphological features. Acridine orange (AO) and propidium iodide (PI) assays were carried out to estimate the incidence of apoptosis. The RT-PCR method was also employed to evaluate the expression of P53, Bax, Bcl-2 and Caspase 3. Results: Exposure to increasing concentrations of TiO2 NPs enhanced overall cell survival of HCT116 cells and reduced the Bcl-2 and Caspase 3 expression while the ratio of Bax/Bcl-2 was down-regulated. TiO2 NPs at 400 and 50 µg/ml concentrations suppressed cell proliferation and induced apoptosis of HT29 cells and also up-regulated P53 and Bax at the mRNA level, enhanced the Bax/Bcl-2 ratio and eventually up-regulated Caspase 3 mRNA. Although, inhibition of cell proliferation in HUVECs was seen at 200 and 400 µg/ml TiO2 NPs, it was not marked. Conclusion: TiO2 NPs have selective bio-effects on exposed cells with dose- and cell-dependent influence on viability. Cell proliferation in HCT116 as a metastatic colorectal cancer cell line appeared to be stimulated via multiple signaling pathways, with promotion of apoptosis in less metastatic cells at 50 and 400 µg/ml concentrations. This was associated with elevated P53, Bax and Caspase 3 mRNA and reduced Bcl-2 expression. However, TiO2 NPs did not exert any apparent significant effects on HUVECs as hyperproliferative angiogenic cells.

Luminescence enhancement in nanocomposite consisting of polyvinyl alcohol incorporated gold nanoparticles and Nile blue 690 perchlorate.

We have experimentally demonstrated that the emission of visible light from the polymer matrix doped with luminescent dye and gold nanoparticles (GNPs) can be enhanced with the use of surface plasmon coupling. GNPs can enhance the luminescence intensity of nearby luminescent dye because of the interactions between the dipole moments of the dye and the surface plasmon field of the GNPs. The electric charge on the GNPs and the distance between GNPs and luminescent dye molecules have a significant effect on the luminescence intensity, and this enhancement depends strongly upon the excitation wavelength of the pumping laser source. In particular, by matching the plasmon frequency of GNPs to the frequency of the laser light source we have observed a strong luminescence enhancement of the nanocomposite consisting of GNPs coupled with luminescent dye Nile blue 690 perchlorate. This ability of controlling luminescence can be beneficially used in developing contrast agents for highly sensitive and specific optical sensing and imaging. This opens new possibilities for plasmonic applications in the solar energy field.

Investigation of the Mg-HCO3⁻-ATPase activity of thyroid tissue cells under various pathologies.

Distribution of the Mg²âº-dependent, HCO3⁻-stimulated ATPase (HCO3⁻-ATPase) was investigated in sub-cellular fractions of the tissue of the human thyroid gland. It was found that especially high enzymatic activity is characteristic of mitochondria and the endoplasmic reticulum. Correlation between various pathologies of the thyroid and HCO3⁻-ATPase activity was demonstrated. The activity was evaluated using the difference between active and passive ATPase. During development of the various pathologies, the sub-cellular fractions of the gland showed uneven alterations of HCO3⁻-ATPase activity. Intriguingly, the enzyme kinetic parameters are also changed, i.e. in the different pathologies both enzyme activity and its affinity towards bicarbonate ions are altered. This raises the question whether pathology is caused by, or results in, changes in the enzyme at the molecular level.

Long-lasting stress, quantitative changes in nitric oxide concentration and functional state of brain mitochondria.

We have studied biochemical changes occurring in brain cell mitochondria of white rats on the background of stress induced by 30-day isolation and disruption of circadian rhythms. It was ascertained that due to long-lasting stress, there occurs activation of oxidative processes in mitochondria as well as inhibition of anti-oxidant system activity, causing development of energy deficiency in brain cells. The above-mentioned biochemical processes become the reason for activation and opening of the mitochondrial permeability transition pore (MPTP), which, in its turn, signals the start of neuroapoptosis and various neurodegenerative processes.

Functional state of rat cardiomyocytes and blood antioxidant system under psycho-emotional stress.

We studied the functionality of the antioxidant system in laboratory rat cardiomyocytes and blood under psycho-emotional stress. It was found that 40-day isolation and violation of diurnal cycle among the animals were accompanied by the intensification of lipid peroxidation process and marked with a reduced activity of antioxidant system enzymes, such as catalase and superoxide dismutase activity. The results suggested that psycho-emotional stress was accompanied by oxidative stress, causing a reduction in the intensity of energy metabolism in cardiomyocytes, which was further strengthened by the fact that the activity of the enzymes involved in ATP synthesis in mitochondria was reduced. Based on the results, we proposed that psychological stress is one of the factors contributing to the development of various cardiac diseases.

Social isolation in rats inhibits oxidative metabolism, decreases the content of mitochondrial K-Ras and activates mitochondrial hexokinase.

Recent observations have suggested that Ras signaling includes combinations of extracellular-signal-regulated Ras activation at the plasma membrane and endomembranes, and translocation of Ras from the plasma membrane to intracellular compartments. In this study we have shown that social isolation of rat decreases the content of Bcl-2-associated K-Ras in hippocampal mitochondria, whereas the amount of H-Ras is increased in the microsomal fraction. Furthermore, we have found that galectin 1, a binding partner of activated Ras, was increased in the soluble fractions. The redistribution of Ras isoforms was accompanied by acceleration in mitochondrial hexokinase and inhibition of mitochondrial aconitase, succinate dehydrogenase, and creatine kinase, whereas the activity of aldolase, as well as cytoplasmic creatine kinase was not changed. Our data suggest that inhibition of mitochondrial oxidative metabolism by reactive oxygen species (ROS) and compensatory elevation of glycolysis in hippocampus occurs during social isolation of rats and Ras trafficking could play an important role in switching of impaired oxidative phosphorylation to anaerobic glycolysis.