Replication timing maintains the global epigenetic state in human cells

ACKNOWLEDGMENTS We thank R. Didier and B. Alexander of the FSU Flow Cytometry and Confocal Microscopy Facilities for their help with flow cytometry and fluorescence-activated cell sorting for this project. Thanks to A. Brown of the FSU Biological Science Core Labs and to Y. Yang and C. Vied of the FSU Translational Labs. Thanks to S. R. Westermann of SCIGRAPHIX for generating the model figure. Thanks to B. van Steensel, J. Phillips-Cremins, and P. Fraser for critical reading of the manuscript. Funding: This work was supported by NIH grant GM083337 to D.M.G., GM035463 to V.G.C., and GM085354 to D.M.G., S.D., and V.G.C. D.L. is supported by the Hong Kong Research Grant Council (ECS 26104216). T.B. is supported by the William C. and Joyce C. O’Neil Charitable Trust, Memorial Sloan Kettering Single Cell Sequencing InitiativePeer reviewedPostprin

Fully Automated One-Step Synthesis of Single-Transcript TALEN Pairs Using a Biological Foundry

Transcription activator-like effector nuclease (TALEN) is a programmable genome editing tool with wide applications. Since TALENs perform cleavage of DNA as heterodimers, a pair of TALENs must be synthesized for each target genome locus. Conventionally, TALEN pairs are either expressed on separate vectors or synthesized separately and then subcloned to the same vector. Neither approach allows high-throughput construction of TALEN libraries for large-scale applications. Here we present a single-step assembly scheme to synthesize and express a pair of TALENs in a single-transcript format with the help of a P2A self-cleavage sequence. Furthermore, we developed a fully automated platform to custom manufacture TALENs in a versatile biological foundry. 400 pairs of TALENs can be synthesized with over 96.2% success rate at a material cost of $2.1/pair. This platform opens the door to TALEN-based genome-wide studies

Case Report: Severe COVID-19 Pneumonia in a Patient With Relapsed/Refractory Hodgkin's Lymphoma

First identified in China in December 2019, coronavirus disease 2019 (COVID-19) has rapidly evolved into a global pandemic. The presence of haematological malignancies are expected to increase the risk of adverse outcomes from this viral infection due to the immunosuppression brought about by the underlying cancer and the effects of therapy. We present a 55-year-old woman diagnosed with relapsed/refractory Hodgkin's lymphoma (HL) who had been heavily pretreated with multiagent chemotherapy, autologous hematopoietic stem cell transplantation (autoHCT), allogeneic hematopoietic stem cell transplantation (alloHCT) and was complicated with EBV associated posttransplant lymphoproliferative disease (PTLD) and chronic graft-versus-host-disease (GVHD). The patient was recently treated with brentuximab and donor lymphocyte infusion (DLI) for relapse after alloHCT. She suffered from severe COVID-19 pneumonia and eventually succumbed to acute respiratory distress syndrome (ARDS) and multiorgan failure. Of note, this is the first reported case of COVID-19 in a HL patient who was being treated with brentuximab for relapse after alloHCT

PROTECTIVE EFFECTS OF GINKGO BILOBA EXTRACT AGAINST CISPLATIN OTOTOXICITY

Cisplatin is one of the most common agents employed in standard treatments of a variety of malignant tumors. However, its clinical application is limited because of serious and sometimes irreversible side effects, that include ototoxicity. Upon cisplatin treatment, several areas of the cochlea are damaged, including outer hair cells in the organ of Corti, spiral ganglion and the stria vascularis. Notwithstanding extensive research, the presently available treatments to prevent ototoxicity are scarcely effective. Cisplatin is thought to interfere with production of endogenous antioxidants that protect inner ear against reactive oxygen species (ROS). Outer hair cells are the most sensitive to ROS damage which can lead to apoptosis: strategies of chemoprevention include the administration of antioxidants to protect hair cells at an early stage in the ototoxic pathways. In this study we evaluated the protective effect of a nutraceutical product compounds with antioxidant activity (ACUVAL®, Scharper Healthcare, Italy). Ginkgo biloba is known for antioxidant properties and for this reason we tested by cytofluorimetry the otoprotective effects of a Ginkgo biloba extract (Ginkgoselect®) against cisplatin-induced toxicity in a mouse inner ear cell line (OCk3). The results support the hypothesis that the pre-treatment with Ginkgo biloba extract (50-100 µg/ml) is able to protect OCk3 against cisplatin-induced toxicity. We are presently investigating the effects of the same extract in rat inner ear organ cultures