Semispecific TPPII inhibitor Ala-Ala-Phe-chloromethylketone (AAF-cmk) displays cytotoxic activity by induction of apoptosis, autophagy and protein aggregation in U937 cells

Introduction. The main component of extralysosomal proteolysis is the ubiquitin-proteasome system (UPS), which is supplemented by tripeptidyl peptidase II (TPPII). That system is a target for anticancer strategies by using proteasome inhibitors. Data from several studies on leukemic cells share evidence for the beneficial and potential role of TPPII in cell survivability. Therefore, the aim of this work was to analyze the effect of AAF-cmk, a membrane permeable semi-specific TPPII inhibitor, on human monocytic leukemic cells U937 for translational research. Material and methods. We studied the viability of U937 cells incubated with AAF-cmk using tetrazolium salt reduction assay (MTT) and apoptosis induction by assessing caspase activation by Western blotting and Annexin V binding assays. Transmission electron microscopy (TEM), a gold standard for apoptosis and autophagy detection, was used to assess the ultrastructure of U937 cells. Results. Incubation of cells with AAF-cmk reduced their viability and induced apoptosis by intrinsic pathway. In groups treated with AAF-cmk, activation of caspases 9 and 3 was observed and caspase inhibition by zVDA restored cell viability. TEM revealed the presence of ultrastructural features of apoptosis and authophagy. Moreover, we identified two types of protein aggregates. The first one was found in close proximity to the endoplasmic reticulum (ER) and corresponds to Aggresome-Like Structure (ALIS); however, the second novel type of aggregate was not related to ER elements, but rather to free cytosolic ribosomes. This type did not correspond to the aggresome neither in localization nor the structure, thus we referred these aggregates as ALiSNER (Aggresome-Like Structure Not Associated With the ER). Conclusions. Our results provide novel and important findings about the role of TPPII in protein homeostasis and cell survival. Since semispecific TPPII inhibitor AAF-cmk displays cytotoxic activity against leukemic U937 cells in vitro it can be considered as a potential anticancer agent

Extremely Low-Frequency Magnetic Field Enhances the Therapeutic Efficacy of Low-Dose Cisplatin in the Treatment of Ehrlich Carcinoma

e present study examines the therapeutic efficacy of the administration of low-dose cisplatin (cis) followed by exposure to extremely low-frequency magnetic �eld (ELF-MF), with an average intensity of 10 mT, on Ehrlich carcinoma in vivo. e cytotoxic and genotoxic actions of this combination were studied using comet assay, mitotic index (MI), and the induction of micronucleus (MN). Moreover, the inhibition of tumor growth was also measured. Treatment with cisplatin and ELF-MF (group A) increased the number of damaged cells by 54% compared with 41% for mice treated with cisplatin alone (group B), 20% for mice treated by exposure to ELF-MF (group C), and 9% for the control group (group D). Also the mitotic index decreased signi�cantly for all treated groups ( ). e decrement percent for the treated groups (A, B, and C) were 70%,65%, and 22%, respectively, compared with the control group (D). Additionally, the rate of tumor growth at day 12 was suppressed signi�cantly ( ) for groups A, B, and C with respect to group (D). ese results suggest that ELF-MF enhanced the cytotoxic activity of cisplatin and potentiate the bene�t of using a combination of low-dose cisplatin and ELF-MF in the treatment of Ehrlich carcinoma

Influence of infrared radiation on the electrical characteristics of the surface-barrier nanostructures based on MBE HgCdTe

Impact of illumination on the admittance of the MIS structures based on MBE Hg1-xCdxTe with graded-gap layers and single quantum wells was investigated. It is shown that for HgCdTe-based nanostructures the illumination greatly affects the capacitance and conductance dependencies. The capacitance-voltage characteristics exhibit a low-frequency behavior, which is associated with a decrease in the differential resistance of the space charge region. Especially informative illumination exposure is in the study of deep traps in n-HgCdTe (x=0.21-0.23) without graded-gap layer. Illumination leads to the low-frequency behavior of capacitance-voltage characteristics of MIS structures based on p-HgCdTe with HgTe single quantum well in the active region, and maximums in the voltage dependences do not appear

Graphene coating obtained in a cold-wall CVD process on the Co-Cr Alloy (L-605) for medical applications

Graphene coating on the cobalt-chromium alloy was optimized and successfully carried out by a cold-wall chemical vapor deposition (CW-CVD) method. A uniform layer of graphene for a large area of the Co-Cr alloy (discs of 10 mm diameter) was confirmed by Raman mapping coated area and analyzing specific G and 2D bands; in particular, the intensity ratio and the number of layers were calculated. The effect of the CW-CVD process on the microstructure and the mor-phology of the Co-Cr surface was investigated by scanning X-ray photoelectron microscope (SPEM), atomic force microscopy (AFM), scanning electron microscopy (SEM), and energy dispersive X-ray spectroscopy (EDS). Nanoindentation and scratch tests were performed to determine mechanical properties of Co-Cr disks. The results of microbiological tests indicate that the studied Co-Cr alloys covered with a graphene layer did not show a pro-coagulant effect. The obtained results confirm the possibility of using the developed coating method in medical applications, in particular in the field of cardiovascular diseases

Steroids and bronchodilators for acute bronchiolitis in the first two years of life: systematic review and meta-analysis

Objective To evaluate and compare the efficacy and safety of bronchodilators and steroids, alone or combined, for the acute management of bronchiolitis in children aged less than 2 years

Synthesis of bifunctional thiophenes via Fiesselmann condensation of ynone trifluoroborate salts

Ynone trifluoroborate salts undergo a base-promoted condensation reaction with alkylthiols to generate thiophene boronates with complete regiocontrol. The products are isolated in high yield and can be further derivatized through conventional C-B bond functionalization reactions