RAD54L2 counters TOP2-DNA adducts to promote genome stability.

The catalytic cycle of topoisomerase 2 (TOP2) enzymes proceeds via a transient DNA double-strand break (DSB) intermediate termed the TOP2 cleavage complex (TOP2cc), in which the TOP2 protein is covalently bound to DNA. Anticancer agents such as etoposide operate by stabilizing TOP2ccs, ultimately generating genotoxic TOP2-DNA protein cross-links that require processing and repair. Here, we identify RAD54 like 2 (RAD54L2) as a factor promoting TOP2cc resolution. We demonstrate that RAD54L2 acts through a novel mechanism together with zinc finger protein associated with tyrosyl-DNA phosphodiesterase 2 (TDP2) and TOP2 (ZATT/ZNF451) and independent of TDP2. Our work suggests a model wherein RAD54L2 recognizes sumoylated TOP2 and, using its ATPase activity, promotes TOP2cc resolution and prevents DSB exposure. These findings suggest RAD54L2-mediated TOP2cc resolution as a potential mechanism for cancer therapy resistance and highlight RAD54L2 as an attractive candidate for drug discovery.

Profile of the multicenter cohort of the German Cancer Consortium's Clinical Communication Platform.

Treatment concepts in oncology are becoming increasingly personalized and diverse. Successively, changes in standards of care mandate continuous monitoring of patient pathways and clinical outcomes based on large, representative real-world data. The German Cancer Consortium's (DKTK) Clinical Communication Platform (CCP) provides such opportunity. Connecting fourteen university hospital-based cancer centers, the CCP relies on a federated IT-infrastructure sourcing data from facility-based cancer registry units and biobanks. Federated analyses resulted in a cohort of 600,915 patients, out of which 232,991 were incident since 2013 and for which a comprehensive documentation is available. Next to demographic data (i.e., age at diagnosis: 2.0% 0-20 years, 8.3% 21-40 years, 30.9% 41-60 years, 50.1% 61-80 years, 8.8% 81+ years; and gender: 45.2% female, 54.7% male, 0.1% other) and diagnoses (five most frequent tumor origins: 22,523 prostate, 18,409 breast, 15,575 lung, 13,964 skin/malignant melanoma, 9005 brain), the cohort dataset contains information about therapeutic interventions and response assessments and is connected to 287,883 liquid and tissue biosamples. Focusing on diagnoses and therapy-sequences, showcase analyses of diagnosis-specific sub-cohorts (pancreas, larynx, kidney, thyroid gland) demonstrate the analytical opportunities offered by the cohort's data. Due to its data granularity and size, the cohort is a potential catalyst for translational cancer research. It provides rapid access to comprehensive patient groups and may improve the understanding of the clinical course of various (even rare) malignancies. Therefore, the cohort may serve as a decisions-making tool for clinical trial design and contributes to the evaluation of scientific findings under real-world conditions.

Clinical prospects of WRN inhibition as a treatment for MSI tumours.

The discovery of synthetic lethal interactions with genetic deficiencies in cancers has highlighted several candidate targets for drug development, with variable clinical success. Recent work has unveiled a promising synthetic lethal interaction between inactivation/inhibition of the WRN DNA helicase and tumours with microsatellite instability, a phenotype that arises from DNA mismatch repair deficiency. While these and further studies have highlighted the therapeutic potential of WRN inhibitors, compounds with properties suitable for clinical exploitation remain to be described. Furthermore, the complexities of MSI development and its relationship to cancer evolution pose challenges for clinical prospects. Here, we discuss possible paths of MSI tumour development, the viability of WRN inhibition as a strategy in different scenarios, and the necessary conditions to create a roadmap towards successful implementation of WRN inhibitors in the clinic.

Study of the Properties of a Biodegradable Polymer Filled with Different Wood Flour Particles.

Lignocellulosic wood flour particles with three different sizes were used to reinforce Solanyl® type bioplastic in three compositions (10, 20, and 30 wt.%) and further processed by melt-extrusion and injection molding to simulate industrial conditions. The wood flour particles were morphologically and granulometric analyzed to evaluate their use as reinforcing filler. The Fuller method on wood flour particles was successfully applied and the obtained results were subsequently corroborated by the mechanical characterization. The rheological studies allowed observing how the viscosity was affected by the addition of wood flour and to recover information about the processing conditions of the biocomposites. Results suggest that all particles can be employed in extrusion processes (shear rate less than 1000 s-1). However, under injection molding conditions, biocomposites with high percentages of wood flour or excessively large particles may cause an increase in defective injected-parts due to obstruction of the gate in the mold. From a processing point of view and based on the biocomposites performance, the best combination resulted in Solanyl® type biopolymer reinforced with wood flour particles loaded up to 20 wt.% of small and medium particles size. The obtained biocomposites are of interest for injected molding parts for several industrial applications.

Neuromarketing Applied to Educational Toy Packaging.

This research work comes in response to the question of which aspects are more relevant for consumers when purchasing educational toys as opposed to other toys that are focused solely on leisure. This empirical research focuses on an educational toy distributed in Spain by the Educa brand (Conector family, reference "I learn English"), which is the product best-selling product of its brand in this area, and analyses how consumers make decisions concerning this product in relation to other products designed by competitors. The research looks into customer reactions while looking at these products, measuring brain activity generated by different aspects of product design and its influence on choice. The aim of the present study was to propose a model that optimizes the design of educational toy packaging. Through the use of neuromarketing techniques -attention through eye tracking, and emotion using galvanic skin response- as well as qualitative research techniques, the objective of this research is to determine the motivations in the processes of buying educational toys. The packaging design elements analyzed are brand, product family, toy name, recommended age, game image, number of questions/topics, and additional texts. The results suggest that the most important elements are the graphic details of the packaging, obtaining a perception of a higher educational level as more questions are addressed by the game. The simultaneous combination of qualitative techniques monitored with galvanic skin response (neuro-qualitative study) allows additional conclusions to be aligned with the end user of the product, including a prominent social component when the product is purchased as a gift for a third party.

Parallel CRISPR-Cas9 screens clarify impacts of p53 on screen performance.

CRISPR-Cas9 genome engineering has revolutionised high-throughput functional genomic screens. However, recent work has raised concerns regarding the performance of CRISPR-Cas9 screens using TP53 wild-type human cells due to a p53-mediated DNA damage response (DDR) limiting the efficiency of generating viable edited cells. To directly assess the impact of cellular p53 status on CRISPR-Cas9 screen performance, we carried out parallel CRISPR-Cas9 screens in wild-type and TP53 knockout human retinal pigment epithelial cells using a focused dual guide RNA library targeting 852 DDR-associated genes. Our work demonstrates that although functional p53 status negatively affects identification of significantly depleted genes, optimal screen design can nevertheless enable robust screen performance. Through analysis of our own and published screen data, we highlight key factors for successful screens in both wild-type and p53-deficient cells.

The invention of CRISPR-Cas9 genome editing has unlocked a greater understanding of the human genome. Researchers can use this system to make targeted cuts in any gene in the genome, forcing the cell to perform a rapid repair at the cut site. These repairs often introduce mutations into the damaged area, adding or removing DNA letters and disrupting the gene. This allows researchers to study what happens to cells when specific genes are missing, which can help to uncover what each gene is for. One of the most comprehensive ways to use this technique is to perform a CRISPR-Cas9 screen, which disrupts each gene in the genome one by one. For a CRISPR-Cas9 screen to work well, a cell needs to survive the cuts to its genome. But there is a crucial gene that can stop this happening. Often described as the 'guardian of the genome', this gene codes for a protein called p53, a tumour suppressor that helps to stop a cell turning cancerous when its DNA becomes damaged. This protein activates when the cell senses a cut in its genetic material and can kill the cell if it fails to make a successful repair. Recent work has shown that the presence of a working copy of the gene for the p53 protein might limit the ability of CRISPR-Cas9 to edit genes. But the evidence was inconclusive. So, Bowden, Morales-Juarez et al. performed two parallel CRISPR-Cas9 screens in human cells with and without p53 to find out more. This revealed that CRISPR-Cas9 can inactivate genes in both normal cells and cells lacking the p53 protein, but that it works better in cells without p53. This was because, when p53 was active, the cells initiated a protective response against the CRISPR-Cas9 cuts. This changed the patterns of genes successfully inactivated by the screen, but it did not make the results unusable. Careful experimental design and thorough data analysis made it possible to get useful results even in cells with functional p53 protein. The gene for p53 has mutations in around half of human cancers. So, understanding how it affects CRISPR-Cas9 screens could influence the design of future experiments. It is possible that the effects of the p53 protein could vary from cell type to cell type, and with different p53 mutations. Comparisons like the one performed here could help to further unpick how the cell's DNA repair systems might interfere with future CRISPR experiments.

Equilibrium clustering of colloidal particles at an oil/water interface due to competing long-range interactions.

HYPOTHESIS: Colloids at fluid interfaces organize according to inter-particle interactions. The main contributions to an effective interaction potential are expected to be electrostatic dipole-dipole repulsion and capillary attraction due to fluid interface deformation. When these interactions are weak, a secondary minimum in the particle pair interaction potential is expected. EXPERIMENTS: Clean bare silica particles were deposited at an oil/water interface and their organization as well as dynamics were observed under a light microscope and analyzed in terms of radial distribution function and mean squared displacement. FINDINGS: Weak long-range competing interactions between colloids at an oil/water interface result in cluster formation. The clusters have a liquid-like structure and grow with increasing particle packing fraction. System 'ergodicity' suggests near-equilibrium assembly, which is confirmed by free particle dynamics outside the clusters. The interplay between dipole-dipole repulsion and capillary attraction responsible for the cluster formation is reflected in a secondary minimum of the effective interaction potential predicted theoretically but inaccessible experimentally from collective particle properties prior to this work.

Automated Reports for Monitoring and Improving Data Quality in a Translational Research Network.

Standardised, automated quality reports were generated at three pilot locations of the decentralized translational research network DKTK with separated local data warehouses (LDW), for assessing syntactic conformity against common data element definitions deposited in a central metadata repository (MDR). Deviations in the LDW were categorised, and locally corrected. Comparisons of reports from two time points confirm a major improvement in data quality in terms of syntactic conformity, an essential prerequisite for network-wide data integration.

Antidiarrheal, vasorelaxant, and neuropharmacological actions of the diterpene tilifodiolide.

Salvia tiliifolia is used in folk medicine as a relaxant agent and for the treatment of diarrhea and neurodegenerative diseases. Tilifodiolide (TFD) is a diterpene obtained from this plant. The purpose of this work was to evaluate the antidiarrheal, vasorelaxant, and neuropharmacological actions of TFD. These effects were selected based on the folk medicinal use of S. tiliifolia. The antidiarrheal activity of 1-50 mg/kg p.o. TFD was assessed with the castor oil related tests. The vasorelaxant effect of TFD (0.9-298 µM) was performed with smooth muscle tissues from rats, and its mechanism of action was evaluated using different inhibitors. The sedative, anxiolytic, and antidepressant effects of 1-100 mg/kg TFD were assessed. The possible mechanisms of action of the anxiolytic and antidepressant effects of TFD were evaluated using inhibitors. TFD exhibited antidiarrheal (ED50 = 10.62 mg/kg) and vasorelaxant (EC50 = 48 ± 3.51 µM) effects. The coadministration of TFD with N(ω)-nitro-L-arginine methyl ester (L-NAME) or 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), reverted the vasorelaxant action showed by TFD alone. TFD exerted anxiolytic actions (ED50 = 20 mg/kg) in the cylinder exploratory test, whereas TFD (50 mg/kg) showed antidepressant actions in the tail suspension test by 44%. The pretreatment with 2 mg/kg flumazenil partially reverted the anxiolytic actions of TFD, whereas the pretreatment with 1 mg/kg yohimbine abolished the antidepressant effects of TFD. In summary, TFD exerted antidiarrheal activity by decreasing the intestinal fluid accumulation and vasorelaxant effects mediated by nitric oxide and cyclic guanosine monophosphate. TFD showed anxiolytic and antidepressant effects by the partial involvement of gamma-Aminobutyric acid (GABA) receptors and the possible participation of α2-adrenoreceptors, respectively.