Reconstruction of a Segment of the UNESCO World Heritage Hadrian’s Villa Tunnel Network by Integrated GPR, Magnetic–Paleomagnetic, and Electric Resistivity Prospections

The authors are grateful to the Director of the Villa Adriana and Villa d’Este, Andrea Bruciati, for kindly allowing us to survey the archaeological area and to Benedetta Adembri for facilitating the research on site. We are also grateful to Francesco Ferruti and the students that helped us in the data acquisition. Finally, we thank Alessandro Bertani for his help in the acquisition and formal analysis of aerial photogrammetry data. This paper also benefited from four accurate reviews that allowed us to improve the manuscript.The following are available online at https://zenodo.org/record/3351757#.XVIUHdIRWUl, Figure S1: Local reference frames used for the acquisition of GPR data, Figure S2: UAV orthophoto of the study area (Plutonium−Inferi complex) with indication of the excavated areas, Tables S1–S13: Relevant migrated and unmigrated GPR profiles for areas 1 through 13, Table S10A, transversal migrated and unmigrated GPR profiles for Areas 10.Hadrian’s Villa is an ancient Roman archaeological site built over an ignimbritic tuff and characterized by abundant iron oxides, strong remnant magnetization, and elevated magnetic susceptibility. These properties account for the high-amplitude magnetic anomalies observed in this site and were used as a primary tool to detect deep archaeological features consisting of air-filled and soil-filled cavities of the tuff. An integrated magnetic, paleomagnetic, radar, and electric resistivity survey was performed in the Plutonium-Inferi sector of Hadrian’s Villa to outline a segment of the underground system of tunnels that link different zones of the villa. A preliminary paleomagnetic analysis of the bedrock unit and a high-resolution topographic survey by aerial photogrammetry allowed us to perform a computer-assisted modelling of the observed magnetic anomalies, with respect to the archaeological sources. The intrinsic ambiguity of this procedure was reduced through the analysis of ground penetrating radar and electric resistivity profiles, while a comprehensive picture of the buried archaeological features was built by integration of the magnetization model with radar amplitude maps. The final subsurface model of the Plutonium-Inferi complex shows that the observed anomalies are mostly due to the presence of tunnels, skylights, and a system of ditches excavated in the tuff.This research was funded by the Università degli Studi di Camerino, grants FAR Schettino 2016–2018 and FAR Pierantoni 2016–2018, and by the University of Oxford, Eugene Ludwig Fund, New College

Combination Therapies Targeting Alk-Aberrant Neuroblastoma in Preclinical Models.

BACKGROUND: ALK activating mutations are identified in approximately 10% of newly diagnosed neuroblastomas and ALK amplifications in a further 1-2% of cases. Lorlatinib, a third generation ALK inhibitor, will soon be given alongside induction chemotherapy for children with ALK-aberrant neuroblastoma. However, resistance to single agent treatment has been reported and therapies that improve the response duration are urgently required. We studied the preclinical combination of lorlatinib with chemotherapy, or with the MDM2 inhibitor, idasanutlin, as recent data has suggested that ALK inhibitor resistance can be overcome through activation of the p53-MDM2 pathway. AIMS: To study the preclinical activity of ALK inhibitors alone and combined with chemotherapy or idasanutlin. METHODS: We compared different ALK inhibitors in preclinical models prior to evaluating lorlatinib in combination with chemotherapy or idasanutlin. We developed a triple chemotherapy (CAV: cyclophosphamide, doxorubicin and vincristine) in vivo dosing schedule and applied this to both neuroblastoma genetically engineered mouse models (GEMM) and patient derived xenografts (PDX). RESULTS: Lorlatinib in combination with chemotherapy was synergistic in immunocompetent neuroblastoma GEMM. Significant growth inhibition in response to lorlatinib was only observed in the ALK-amplified PDX model with high ALK expression. In this PDX lorlatinib combined with idasanutlin resulted in complete tumor regression and significantly delayed tumor regrowth. CONCLUSION: In our preclinical neuroblastoma models, high ALK expression was associated with lorlatinib response alone or in combination with either chemotherapy or idasanutlin. The synergy between MDM2 and ALK inhibition warrants further evaluation of this combination as a potential clinical approach for children with neuroblastoma

Sensitivity of Global Translation to mTOR Inhibition in REN Cells Depends on the Equilibrium between eIF4E and 4E-BP1

Initiation is the rate-limiting phase of protein synthesis, controlled by signaling pathways regulating the phosphorylation of translation factors. Initiation has three steps, 43S, 48S and 80S formation. 43S formation is repressed by eIF2α phosphorylation. The subsequent steps, 48S and 80S formation are enabled by growth factors. 48S relies on eIF4E-mediated assembly of eIF4F complex; 4E-BPs competitively displace eIF4E from eIF4F. Two pathways control eIF4F: 1) mTORc1 phosphorylates and inactivates 4E-BPs, leading to eIF4F formation; 2) the Ras-Mnk cascade phosphorylates eIF4E. We show that REN and NCI-H28 mesothelioma cells have constitutive activation of both pathways and maximal translation rate, in the absence of exogenous growth factors. Translation is rapidly abrogated by phosphorylation of eIF2α. Surprisingly, pharmacological inhibition of mTORc1 leads to the complete dephosphorylation of downstream targets, without changes in methionine incorporation. In addition, the combined administration of mTORc1 and MAPK/Mnk inhibitors has no additive effect. The inhibition of both mTORc1 and mTORc2 does not affect the metabolic rate. In spite of this, mTORc1 inhibition reduces eIF4F complex formation, and depresses translocation of TOP mRNAs on polysomes. Downregulation of eIF4E and overexpression of 4E-BP1 induce rapamycin sensitivity, suggesting that disruption of eIF4F complex, due to eIF4E modulation, competes with its recycling to ribosomes. These data suggest the existence of a dynamic equilibrium in which eIF4F is not essential for all mRNAs and is not displaced from translated mRNAs, before recycling to the next

Concepts of epigenetics in prostate cancer development

Substantial evidence now supports the view that epigenetic changes have a role in the development of human prostate cancer. Analyses of the patterns of epigenetic alteration are providing important insights into the origin of this disease and have identified specific alterations that may serve as useful diagnostic and prognostic biomarkers. Examination of cancer methylation patterns supports a stem cell origin of prostate cancer. It is well established that methylation of GSTpi is a marker of prostate cancer, and global patterns of histone marking appear to be linked to cancer prognosis with levels of acetylated histones H3K9, H3K18, and H4K12, and of dimethylated H4R3 and H3K4, dividing low-grade prostate cancer (Gleason 6 or less) into two prognostically separate groups. Elevated levels of several components of the polycomb group protein complex, EZH2, BMI1, and RING1, can also act as biomarkers of poor clinical outcome. Many components of the epigenetic machinery, including histone deacetylase (whose expression level is linked to the TMPRSS2: ERG translocation) and the histone methylase EZH2, are potential therapeutic targets. The recent discovery of the role of small RNAs in governing the epigenetic status of individual genes offers exciting new possibilities in therapeutics and chemoprevention