Quercetin suppresses heat shock-induced nuclear translocation of Hsp72

The effect of quercetin and heat shock on the Hsp72 level and distribution in HeLa cells was studied by Western blotting, indirect immunofluorescence and immunogold electron microscopy. In control cells and after quercetin treatment, Hsp72 was located both in the cytoplasm and in the nucleus in comparable amounts. After hyperthermia, the level of nuclear Hsp72 raised dramatically. Expression of Hsp72 in cytoplasm was also higher but not to such extent as that observed in the nucleus. Preincubation of heated cells with quercetin inhibited strong Hsp72 expression observed after hyperthermia and changed the intracellular Hsp72 distribution. The cytoplasmic level of protein exceeded the nuclear one, especially around the nucleus, where the coat of Hsp72 was noticed. Observations indicating that quercetin was present around and in the nuclear envelope suggested an involvement of this drug in the inhibition of nuclear translocation. Our results indicate that pro-apoptotic activity of quercetin may be correlated not only with the inhibition of Hsp72 expression but also with suppression of its migration to the nucleus

Microstructure and Properties of Hydroxyapatite Coatings Made by Aerosol Cold Spraying–Sintering Technology

Hydroxyapatite is a widely used material used for the bioactivation of an implant’s surface. A promising hydroxyapatite coating approach is the kinetic deposition of powder particles. The possibility of solid-state deposition improvement through the merging of Aerosol Deposition and Low Pressure Cold Spraying techniques is a promising prospect for improving the deposition efficiency and the quality of coatings. The objective of the paper is to study the possibilities of hydroxyapatite coating structure modification through changes in the coating process and post-heat treatment. The novel Aerosol Cold Spraying system joining Low Pressure Cold Spraying and Aerosol Deposition was used for the deposition of coatings. The coating’s post-processing was conducted using two techniques: Spark Plasma Sintering and Pressureless Sintering. The coating’s structure was examined using scanning, transmission, and light microscopy, and X-ray diffraction. Substrate–coating bond strength was assessed using a tensile test. Homogenous buildup using Aerosol Cold Spraying of hydroxyapatite was achieved. Various pores and microcracks were visible in the sprayed coatings. The deposition process and the thermal post-processing did not lead to significant degradation of the hydroxyapatite phase. As a result of the Spark Plasma Sintering and Pressureless Sintering at 800 °C, an increase in tensile adhesion bond strength and crystal size was obtained

The Role of Bcl-2 and Beclin-1 Complex in “Switching” between Apoptosis and Autophagy in Human Glioma Cells upon LY294002 and Sorafenib Treatment

Background: Gliomas are the most malignant tumors of the central nervous system. One of the factors in their high drug resistance is avoiding programmed death (PCD) induction. This is related to the overexpression of intracellular survival pathways: PI3K-Akt/PKB-mTOR and Ras-Raf-MEK-ERK. Apoptosis and autophagy are co-existing processes due to the interactions between Bcl-2 and beclin-1 proteins. Their complex may be a molecular “toggle-switch” between PCD types. The aim of this research was to investigate the role of Bcl-2:beclin-1 complex in glioma cell elimination through the combined action of LY294002 and sorafenib. Methods: Drug cytotoxicity was estimated with an MTT test. The type of cell death was evaluated using variant microscopy techniques (fluorochrome staining, immunocytochemistry, and transmission electron microscopy), as well as the Bcl-2:beclin-1 complex formation and protein localization. Molecular analysis of PCD indicators was conducted through immunoblotting, immunoprecipitation, and ELISA testing. SiRNA was used to block Bcl-2 and beclin-1 expression. Results: The results showed the inhibitors used in simultaneous application resulted in Bcl-2:beclin-1 complex formation and apoptosis becoming dominant. This was accompanied by changes in the location of the tested proteins. Conclusions: “Switching” between apoptosis and autophagy using PI3K and Raf inhibitors with Bcl-2:beclin-1 complex formation opens new therapeutic perspectives against gliomas