Targeting the NPL4 Adaptor of p97/VCP Segregase by Disulfiram as an Emerging Cancer Vulnerability Evokes Replication Stress and DNA Damage while Silencing the ATR Pathway

Research on repurposing the old alcohol-aversion drug disulfiram (DSF) for cancer treatment has identified inhibition of NPL4, an adaptor of the p97/VCP segregase essential for turnover of proteins involved in multiple pathways, as an unsuspected cancer cell vulnerability. While we reported that NPL4 is targeted by the anticancer metabolite of DSF, the bis-diethyldithiocarbamate-copper complex (CuET), the exact, apparently multifaceted mechanism(s) through which the CuET-induced aggregation of NPL4 kills cancer cells remains to be fully elucidated. Given the pronounced sensitivity to CuET in tumor cell lines lacking the genome integrity caretaker proteins BRCA1 and BRCA2, here we investigated the impact of NPL4 targeting by CuET on DNA replication dynamics and DNA damage response pathways in human cancer cell models. Our results show that CuET treatment interferes with DNA replication, slows down replication fork progression and causes accumulation of single-stranded DNA (ssDNA). Such a replication stress (RS) scenario is associated with DNA damage, preferentially in the S phase, and activates the homologous recombination (HR) DNA repair pathway. At the same time, we find that cellular responses to the CuET-triggered RS are seriously impaired due to concomitant malfunction of the ATRIP-ATR-CHK1 signaling pathway that reflects an unorthodox checkpoint silencing mode through ATR (Ataxia telangiectasia and Rad3 related) kinase sequestration within the CuET-evoked NPL4 protein aggregates

Simulation software for electronics

Práce je věnována rešerši simulačních nástrojů pro elektrotechniku a elektroniku. Bude popisovat a srovnávat možnosti několika simulačních nástrojů, které jsou dostupné na trhu. Dále bude dle zvoleného simulačního nástroje ukazovat vytvoření konkrétního elektrotechnické simulace daného elektrotechnického schématu.The work is devoted for background research of simulation tools for electrical engineering and electronic. It will describe and compare several options of simulation tools available on the market. Further will, according to the selected simulation tool, show the creation of a specific electrical simulation of the electrical diagram.Katedra informačních technologiíDokončená práce s úspěšnou obhajobo

Effect of Sepatronium Bromide (YM-155) on DNA Double-Strand Breaks Repair in Cancer Cells

Survivin, as an antiapoptotic protein often overexpressed in cancer cells, is a logical target for potential cancer treatment. By overexpressing survivin, cancer cells can avoid apoptotic cell death and often become resistant to treatments, representing a significant obstacle in modern oncology. A survivin suppressor, an imidazolium-based compound known as YM-155, is nowadays studied as an attractive anticancer agent. Although survivin suppression by YM-155 is evident, researchers started to report that YM-155 is also an inducer of DNA damage introducing yet another anticancer mechanism of this drug. Moreover, the concentrations of YM-155 for DNA damage induction seems to be far lower than those needed for survivin inhibition. Understanding the molecular mechanism of action of YM-155 is of vital importance for modern personalized medicine involving the selection of responsive patients and possible treatment combinations. This review focuses mainly on the documented effects of YM-155 on DNA damage signaling pathways. It summarizes up to date literature, and it outlines the molecular mechanism of YM-155 action in the context of the DNA damage field

Comparison of Capacity Fade for the Constant Current and WLTC Drive Cycle Discharge Modes for Commercial LiFeYPO4 Cells Used in xEV Vehicles

In this paper, capacity fade of LiFeYPO4/graphite commercial cells during 116 cycles under different temperatures is studied. The cells were discharged in two modes, during Drive Cycle (DrC) discharge cycles the cell was discharged with current waveform calculated for example battery electric vehicle (BEV) under WLTC 3b drive cycle conditions, whereas during Constant Current (CC) discharge cycles the cell was discharged with a constant current of the same root mean square of the current, as the WLTC 3b current waveform and with the same depth of discharge. All the cells were charged in constant current/constant voltage mode. Two fresh cells were used for each discharge mode at 25 °C and as the results were similar, only one cell per discharge mode was used at the other temperatures 5 °C and 45 °C. Furthermore, simulation P2D model of calendar and cycle life was calibrated based on experimental data. SoC floating was observed during cycling for both discharge modes, accompanied with slight increase in end discharge voltage and growth of energy efficiency. Concluding the results for 25 °C, not waveform character, but the amount of electric charge in combination with calendar aging has the most effect on the cycle life, which is also proved by the simulation. For 5 °C, the capacity fade is milder for DrC discharge cycles, but simulation results do not prove that, which would demand further investigation. The results for 45 °C are apparently dependent on a higher amount of discharged and charged electric charge and influenced by calendar life, simulated capacity fade corresponds quite well to the experiment. The best State of Health (SoH) simulation results are for temperature 45 °C, RMSE is 0.10% SoH, for the other temperatures RMSE is 0.20 and 0.93% SoH for 25 and 5 °C, respectively

Expression, purification and assembly of soluble multimeric MHC class II–invariant chain complexes

AbstractMajor histocompatibility class (MHC) II molecules are essential for running adaptive immune response. They are produced in the ER and targeted to late endosomes with the help of invariant chain (Ii) trimers. Ii trimerization may be induced by the Ii TM domain. To enable mechanistic and structural studies of MHC class II–Ii assembly, soluble forms of the complexes were expressed. We show that Ii trimerizes in the absence of the transmembrane part, prior to binding of α/β chains. The biochemical analysis supports the suggestion that the MHC class II–Ii complexes are not necessarily trimers of trimers, but that the Ii trimer can also be occupied by one or two MHC class II complexes