Cisplatin sensitivity of testis tumour cells is due to deficiency in interstrand-crosslink repair and low ERCC1-XPF expression

<p>Abstract</p> <p>Background</p> <p>Cisplatin based chemotherapy cures over 80% of metastatic testicular germ cell tumours (TGCT). In contrast, almost all other solid cancers in adults are incurable once they have spread beyond the primary site. Cell lines derived from TGCTs are hypersensitive to cisplatin reflecting the clinical response. Earlier findings suggested that a reduced repair capacity might contribute to the cisplatin hypersensitivity of testis tumour cells (TTC), but the critical DNA damage has not been defined. This study was aimed at investigating the formation and repair of intrastrand and interstrand crosslinks (ICLs) induced by cisplatin in TTC and their contribution to TTC hypersensitivity.</p> <p>Results</p> <p>We observed that repair of intrastrand crosslinks is similar in cisplatin sensitive TTC and resistant bladder cancer cells, whereas repair of ICLs was significantly reduced in TTC. γH2AX formation, which serves as a marker of DNA breaks formed in response to ICLs, persisted in cisplatin-treated TTC and correlated with sustained phosphorylation of Chk2 and enhanced PARP-1 cleavage. Expression of the nucleotide excision repair factor ERCC1-XPF, which is implicated in the processing of ICLs, is reduced in TTC. To analyse the causal role of ERCC1-XPF for ICL repair and cisplatin sensitivity, we over-expressed ERCC1-XPF in TTC by transient transfection. Over-expression increased ICL repair and rendered TTC more resistant to cisplatin, which suggests that ERCC1-XPF is rate-limiting for repair of ICLs resulting in the observed cisplatin hypersensitivity of TTC.</p> <p>Conclusion</p> <p>Our data indicate for the first time that the exceptional sensitivity of TTC and, therefore, very likely the curability of TGCT rests on their limited ICL repair due to low level of expression of ERCC1-XPF.</p

Hydrogel Containing Anti-CD44-Labeled Microparticles, Guide Bone Tissue Formation in Osteochondral Defects in Rabbits

Hydrogels are suitable for osteochondral defect regeneration as they mimic the viscoelastic environment of cartilage. However, their biomechanical properties are not sufficient to withstand high mechanical forces. Therefore, we have prepared electrospun poly-ε-caprolactone-chitosan (PCL-chit) and poly(ethylene oxide)-chitosan (PEO-chit) nanofibers, and FTIR analysis confirmed successful blending of chitosan with other polymers. The biocompatibility of PCL-chit and PEO-chit scaffolds was tested; fibrochondrocytes and chondrocytes seeded on PCL-chit showed superior metabolic activity. The PCL-chit nanofibers were cryogenically grinded into microparticles (mean size of about 500 µm) and further modified by polyethylene glycol–biotin in order to bind the anti-CD44 antibody, a glycoprotein interacting with hyaluronic acid (PCL-chit-PEGb-antiCD44). The PCL-chit or PCL-chit-PEGb-antiCD44 microparticles were mixed with a composite gel (collagen/fibrin/platelet rich plasma) to improve its biomechanical properties. The storage modulus was higher in the composite gel with microparticles compared to fibrin. The Eloss of the composite gel and fibrin was higher than that of the composite gel with microparticles. The composite gel either with or without microparticles was further tested in vivo in a model of osteochondral defects in rabbits. PCL-chit-PEGb-antiCD44 significantly enhanced osteogenic regeneration, mainly by desmogenous ossification, but decreased chondrogenic differentiation in the defects. PCL-chit-PEGb showed a more homogeneous distribution of hyaline cartilage and enhanced hyaline cartilage differentiation

Mechanistic insights into p53‐regulated cytotoxicity of combined entinostat and irinotecan against colorectal cancer cells

Late‐stage colorectal cancer (CRC) is still a clinically challenging problem. The activity of the tumor suppressor p53 is regulated via post‐translational modifications (PTMs). While the relevance of p53 C‐terminal acetylation for transcriptional regulation is well defined, it is unknown whether this PTM controls mitochondrially mediated apoptosis directly. We used wild‐type p53 or p53‐negative human CRC cells, cells with acetylation‐defective p53, transformation assays, CRC organoids, and xenograft mouse models to assess how p53 acetylation determines cellular stress responses. The topoisomerase‐1 inhibitor irinotecan induces acetylation of several lysine residues within p53. Inhibition of histone deacetylases (HDACs) with the class I HDAC inhibitor entinostat synergistically triggers mitochondrial damage and apoptosis in irinotecan‐treated p53‐positive CRC cells. This specifically relies on the C‐terminal acetylation of p53 by CREB‐binding protein/p300 and the presence of C‐terminally acetylated p53 in complex with the proapoptotic BCL2 antagonist/killer protein. This control of C‐terminal acetylation by HDACs can mechanistically explain why combinations of irinotecan and entinostat represent clinically tractable agents for the therapy of p53‐proficient CRC

Integrated real-time control of water reservoirs with deterministic and probabilistic multi-timescale forecasts: Application to the Lake Como

Hydro-meteorological forecasts are more and more easily available with improving skill over longer timescales and with higher spatiotemporal resolutions. Their uncertainties are commonly represented by ensemble prediction systems which are now dominant at the global and continental scale, while short-term deterministic forecasts are still used, especially in some local contexts. For effective water resources management, it is critical to understand how to use the wealth of information provided by different forecast systems over multiple timescales to help meet the water demand of key competing socio-economic sectors, while reducing short-term impacts and bringing the controlled systems to desirable states in the long term. Real-time control schemes of water reservoirs like Model Predictive Control (MPC) can help meet these goals, by providing a flexible framework to use forecasts proactively and satisfy multiple competing objectives while respecting operational constraints. In this study, we propose a new nested multi-stage stochastic MPC framework integrating the use of both deterministic and ensemble hydrological forecasts over multiple timescales from short-term (60 hours) to seasonal (7 months ahead). We demonstrate the performance of this real-time controller for the Lake Como system, located in the Italian Alps, where a large lake is regulated mainly for irrigation supply and flood control. First, seasonal ensemble forecasts are used to solve a Tree-Based MPC (TB-MPC) problem optimising the reservoir management over several months, by adopting a tree structure to summarise the ensemble information including the resolution of uncertainty in time. Second, the decisions identified so far are used to condition daily operations over a month using sub-seasonal probabilistic forecasts (up to 46 days) under the same TB-MPC approach. Third, the decisions for the first few days are then further adapted to optimize operations three days ahead using deterministic short-term forecasts with MPC. The sub-seasonal and seasonal ensemble (re-)forecasts used are those produced by the European Flood Awareness System (EFAS) from the Copernicus Emergency Management Service which uses ensemble meteorological forecasts from the European Centre for Medium-Range Weather Forecasts (ECMWF). While EFAS is uncalibrated for the study area, we apply bias-correction techniques to improve the agreement of forecasts with local observations and allow their use for resolving the water-balance within MPC. The short-term 60-h forecasts come from a locally calibrated hydrological model (TOPKAPI) using deterministic weather forecasts from the COSMO (Consortium for Small Scale Modelling) model. The skill of all these forecasts is assessed, as well as the ensemble spread–error relationship for EFAS at different lead times. To evaluate the value of the forecasts we compare the performance of the real-time MPC controller with different benchmarks including perfect forecasts, climatology, and persistence. Finally, we investigate the link between forecast skill and value for reservoir operation, and we compare the performance of the nested MPC framework integrating multi-timescale forecasts with the MPC using single forecasts

Synovial cysts of the hip

Background: Synovial cysts of the hip are relatively rare lesions comparing to other joints. Patients are usually asymptomatic, but in some cases symptoms such as pain and/or compression of vessels or nerve could be present. Purpose of the study was to define clinical features and optimal management of synovial cyst of the hip joint through an accurate review of the literature. Methods: we present 3 consecutive cases treated with 3 different therapeutic strategies: surgical excision, wait-and-see and needle aspiration. An accurate review of the literature has been performed to identify patients who had been treated for synovial cyst of the hip. Results: due to the rarity of the disease, there are no significant data in literature supporting the gold standard of treatment. Treatment of the synovial cyst depends on their size, symptoms and comorbities. Conclusions: most of the authors recommend surgical treatment for symptomatic synovial cysts and needle aspiration as an option treatment in asymptomatic patients without vessel or nerve compression. In patients that referred symptoms in correspondence with the hip joint, not strictly related with radiograph findings, a CT or MRI examinations should be performed to exclude possible differential diagnosis