TET-mediated DNA hydroxymethylation is negatively influenced by the PARP-dependent PARylation

Background Poly(ADP-ribosyl)ation (PARylation), a posttranslational modification introduced by PARP-1 and PARP-2, has first been implicated in DNA demethylation due to its role in base excision repair. Recent evidence indicates a direct influence of PARP-dependent PARylation on TET enzymes which catalyse hydroxymethylation of DNA—the first step in DNA demethylation. However, the exact nature of influence that PARylation exerts on TET activity is still ambiguous. In our recent study, we have observed a negative influence of PARP-1 on local TET-mediated DNA demethylation of a single gene and in this study, we further explore PARP–TET interplay. Results Expanding on our previous work, we show that both TET1 and TET2 can be in vitro PARylated by PARP-1 and PARP-2 enzymes and that TET1 PARylation negatively affects the TET1 catalytic activity in vitro. Furthermore, we show that PARylation inhibits TET-mediated DNA demethylation at the global genome level in cellulo. Conclusions According to our findings, PARP inhibition can positively influence TET activity and therefore affect global levels of DNA methylation and hydroxymethylation. This gives a strong rationale for future examination of PARP inhibitors' potential use in the therapy of cancers characterised by loss of 5-hydroxymethylcytosine

EpiCRISPR targeted methylation of Arx gene initiates transient switch of mouse pancreatic alpha to insulin-producing cells

Introduction: Beta cell dysfunction by loss of beta cell identity, dedifferentiation, and the presence of polyhormonal cells are main characteristics of diabetes. The straightforward strategy for curing diabetes implies reestablishment of pancreatic beta cell function by beta cell replacement therapy. Aristaless-related homeobox (Arx) gene encodes protein which plays an important role in the development of pancreatic alpha cells and is a main target for changing alpha cell identity. Results: In this study we used CRISPR/dCas9-based epigenetic tools for targeted hypermethylation of Arx gene promoter and its subsequent suppression in mouse pancreatic αTC1-6 cell line. Bisulfite sequencing and methylation profiling revealed that the dCas9-Dnmt3a3L-KRAB single chain fusion constructs (EpiCRISPR) was the most efficient. Epigenetic silencing of Arx expression was accompanied by an increase in transcription of the insulin gene (Ins2) mRNA on 5th and 7th post-transfection day, quantified by both RT-qPCR and RNA-seq. Insulin production and secretion was determined by immunocytochemistry and ELISA assay, respectively. Eventually, we were able to induce switch of approximately 1% of transiently transfected cells which were able to produce 35% more insulin than Mock transfected alpha cells. Conclusion: In conclusion, we successfully triggered a direct, transient switch of pancreatic alpha to insulin-producing cells opening a future research on promising therapeutic avenue for diabetes management. 1 Introductio

Endotracheal stent increased survival length in patients with invasive thymic adenocarcinoma

We reported the first case of inoperative thymic adenocarcinoma successfully palliative treated by the double-stent procedure. In a patient who expressed stridor, computed tomography was done and necrotic mediastinal mass, which protrudes into a trachea, was demonstrated. Fiberoptic bronchoscopy showed tracheal infiltration and 70% stenosis; therefore, surgical resection was inapplicable. Recanalization with repeated argon plasma coagulation and debridement of necrotic mass was performed, followed by placement of the endotracheal stent, radiotherapy, and chemotherapy. After 1 year, the patient developed gastric aspiration and tracheoesophageal fistula; therefore, the esophageal stent was placed. The outcome was lethal, but the placement of endotracheal stent significantly increased a length of survival for the patient with invasive thymic adenocarcinoma

Achieving a Superhydrophobic, Moisture, Oil and Gas Barrier Film Using a Regenerated Cellulose–Calcium Carbonate Composite Derived from Paper Components or Waste

It has been a persistent challenge to develop eco-friendly packaging cellulose film providing the required multiple barrier properties whilst simultaneously contributing to a circular economy. Typically, a cellulosic film made from nanocellulose materials presents severe limitations, such as poor water/moisture resistance and lacking water vapour barrier properties, related primarily to the hydrophilic and hygroscopic nature of cellulose. In this work, alkyl ketene dimer (AKD) and starch, both eco-friendly, non-toxic, cost-effective materials, were used to achieve barrier properties of novel cellulose–calcium carbonate composite films regenerated from paper components, including paper waste, using ionic liquid as solvent. AKD and starch were applied first into the ionic cellulose solution dope mix, and secondly, AKD alone was coated from hot aqueous suspension onto the film surface using a substrate surface precooling technique. The interactions between the AKD and cellulose film were characterised by Fourier-Transform Infrared Spectroscopy (FTIR) and X-ray Diffraction (XRD) showing the formation of a ketone ester structure between AKD and the hydroxyl groups of cellulose. The presence of calcium carbonate particles in the composite was seen to enhance the cellulose crystallinity. The initial high-water vapour and oxygen transmission rates of the untreated base films could be decreased significantly from 2.00 to 0.14 g m−2 d−1, and 3.85 × 102 to 0.45 × 102 cm3 m−2 d−1, respectively. In addition, by applying subsequent heat treatment to the AKD coating, the water contact angle was markedly increased to reach levels of superhydrophobicity (>150°, and roll-off angle < 5°). The resistance to water absorption, grease-permeation, and tensile strength properties were ultimately improved by 41.52%, 95.33%, and 127.33%, respectively, compared with those of an untreated pure cellulose film. The resulting regenerated cellulose–calcium carbonate composite-based film and coating formulation can be considered to provide a future bio-based circular economy barrier film, for example, for the packaging, construction and agriculture industries, to complement or replace oil-based plastics.Peer reviewe