RIG-I plays a dominant role in the induction of transcriptional changes in Zika virus-infected cells and protects from virus-induced cell death

The Zika virus (ZIKV) has received much attention due to an alarming increase in cases of neurological disorders including congenital Zika syndrome associated with infection. To date, there is no effective treatment available. An immediate response by the innate immune system is crucial for effective control of the virus. Using CRISPR/Cas9-mediated knockouts in A549 cells, we investigated the individual contributions of the RIG-I-like receptors MDA5 and RIG-I to ZIKV sensing and control of this virus by using a Brazilian ZIKV strain. We show that RIG-I is the main sensor for ZIKV in A549 cells. Surprisingly, we observed that loss of RIG-I and consecutive type I interferon (IFN) production led to virus-induced apoptosis. ZIKV non-structural protein NS5 was reported to interfere with type I IFN receptor signaling. Additionally, we show that ZIKV NS5 inhibits type I IFN induction. Overall, our study highlights the importance of RIG-I-dependent ZIKV sensing for the prevention of virus-induced cell death and shows that NS5 inhibits the production of type I IFN

Infection with a Brazilian isolate of Zika virus generates RIG‐I stimulatory RNA and the viral NS5 protein blocks type I IFN induction and signalling

Zika virus (ZIKV) is a major public health concern in the Americas. We report that ZIKV infection and RNA extracted from ZIKV infected cells potently activated the induction of type I interferons (IFNs). This effect was fully dependent on the mitochondrial antiviral signalling protein (MAVS), implicating RIG‐I‐like receptors (RLRs) as upstream sensors of viral RNA. Indeed, RIG‐I and the related RNA sensor MDA5 contributed to type I IFN induction in response to RNA from infected cells. We found that ZIKV NS5 from a recent Brazilian isolate blocked type I IFN induction downstream of RLRs and also inhibited type I IFN receptor (IFNAR) signalling. We defined the ZIKV NS5 nuclear localization signal and report that NS5 nuclear localization was not required for inhibition of signalling downstream of IFNAR. Mechanistically, NS5 blocked IFNAR signalling by both leading to reduced levels of STAT2 and by blocking phosphorylation of STAT1, two transcription factors activated by type I IFNs. Taken together, our observations suggest that ZIKV infection induces a type I IFN response via RLRs and that ZIKV interferes with this response by blocking signalling downstream of RLRs and IFNAR

VESTA - Very-High-Temperature Heat Aquifer Storage

Energy storage is one of the key challenges of the energy transition. Eight international partners from Germany, Switzerland and the USA address this challenge in the joint project VESTA. Goal of VESTA is the generic development and demonstration of high-temperature storage in the underground. Four pilot sites in the DACH region in various geologies and project phases allow feedback loops between generic scientific investigations and application of new geothermal technologies. Specifically, pilot sites that shall 1) demonstrate HT-ATES technology, 2) evaluate technical and non-technical barriers, 3) support development and implementation by providing techniques and optimized component design, and 4) support agencies with scientific and technical knowledge as a basis for advancing regulatory provisions. With this scientific program, VESTA shall form a technical-economic bases for future operational concepts

Non-isothermal flow in low permeable porous media: a comparison of Richards’ and two-phase flow approaches

The present work compares the performance of two alternative flow models for the simulation of thermal-hydraulic coupled processes in low permeable porous media: non-isothermal Richards and two-phase flow concepts. Both models take vaporization processes into account: however, the Richards model neglects dynamic pressure variations and bulk flow of the gaseous phase. For the comparison of the two approaches first published data from a laboratory experiment is studied involving thermally driven moisture flow in a partially saturated bentonite sample. Then a benchmark test of longer-term thermal-hydraulic behavior in the engineered barrier system of a geological nuclear waste repository is analyzed (DECOVALEX project). It was found that both models can be used to reproduce the vaporization process if the intrinsic permeability is relative high. However, when a thermal-hydraulic coupled problem has the same low intrinsic permeability for both the liquid and the gas phase, only the two-phase flow approach provides reasonable results

A numerical manifold method model for analyzing fully coupled hydro-mechanical processes in porous rock masses with discrete fractures

In this study, a numerical manifold method (NMM) model was developed for fully coupled analysis of hydro-mechanical (HM) processes in porous rock masses with discrete fractures. Using an NMM two-cover-mesh system of mathematical and physical covers, fractures are conveniently discretized by dividing the mathematical cover along fracture traces to physical cover, resulting in a discontinuous model on a non-conforming mesh. In this model, discrete fracture deformation (e.g. open and slip) and fracture fluid flow within a permeable and deformable porous rock matrix are rigorously considered. For porous rock, direct pore-volume coupling was modeled based on an energy-work scheme. For mechanical analysis of fractures, a fracture constitutive model for mechanically open states was introduced. For fluid flow in fractures, both along-fracture and normal-to-fracture fluid flow are modeled without introducing additional degrees of freedom. When the mechanical aperture of a fracture is changing, its hydraulic aperture and hydraulic conductivity is updated. At the same time, under the effect of coupled deformation and fluid flow, the contact state may dynamically change, and the corresponding contact constraint is updated each time step. Therefore, indirect coupling is realized under stringent considerations of coupled HM effects and fracture constitutive behavior transfer dynamically. To verify the new model, examples involving deformable porous media containing a single and two sets of fractures were designed, showing good accuracy. Last, the model was applied to analyze coupled HM behavior of fractured porous rock domains with complex fracture networks under effects of loading and injection

Status of the TOUGH-FLAC simulator and recent applications related to coupled fluid flow and crustal deformations

This paper presents recent advancement in and applications of TOUGH-FLAC, a simulator for multiphase fluid flow and geomechanics. The TOUGH-FLAC simulator links the TOUGH family multiphase fluid and heat transport codes with the commercial FLAC{sup 3D} geomechanical simulator. The most significant new TOUGH-FLAC development in the past few years is a revised architecture, enabling a more rigorous and tight coupling procedure with improved computational efficiency. The applications presented in this paper are related to modeling of crustal deformations caused by deep underground fluid movements and pressure changes as a result of both industrial activities (the In Salah CO{sub 2} Storage Project and the Geysers Geothermal Field) and natural events (the 1960s Matsushiro Earthquake Swarm). Finally, the paper provides some perspectives on the future of TOUGH-FLAC in light of its applicability to practical problems and the need for high-performance computing capabilities for field-scale problems, such as industrial-scale CO{sub 2} storage and enhanced geothermal systems. It is concluded that despite some limitations to fully adapting a commercial code such as FLAC{sup 3D} for some specialized research and computational needs, TOUGH-FLAC is likely to remain a pragmatic simulation approach, with an increasing number of users in both academia and industry

Immunogenomics of colorectal cancer response to checkpoint blockade: analysis of the KEYNOTE 177 trial and validation cohorts

BACKGROUND & AIMS: Colorectal cancer (CRC) shows variable response to immune checkpoint blockade, which can only partially be explained by high tumor mutational burden (TMB). We conducted an integrated study of the cancer tissue and associated tumor microenvironment (TME) from patients treated with pembrolizumab (KEYNOTE 177 clinical trial) or nivolumab to dissect the cellular and molecular determinants of response to anti- programmed cell death 1 (PD1) immunotherapy. METHODS: We selected multiple regions per tumor showing variable T-cell infiltration for a total of 738 regions from 29 patients, divided into discovery and validation cohorts. We performed multiregional whole-exome and RNA sequencing of the tumor cells and integrated these with T-cell receptor sequencing, high-dimensional imaging mass cytometry, detection of programmed death-ligand 1 (PDL1) interaction in situ, multiplexed immunofluorescence, and computational spatial analysis of the TME. RESULTS: In hypermutated CRCs, response to anti-PD1 immunotherapy was not associated with TMB but with high clonality of immunogenic mutations, clonally expanded T cells, low activation of Wnt signaling, deregulation of the interferon gamma pathway, and active immune escape mechanisms. Responsive hypermutated CRCs were also rich in cytotoxic and proliferating PD1+CD8 T cells interacting with PDL1+ antigen-presenting macrophages. CONCLUSIONS: Our study clarified the limits of TMB as a predictor of response of CRC to anti-PD1 immunotherapy. It identified a population of antigen-presenting macrophages interacting with CD8 T cells that consistently segregate with response. We therefore concluded that anti-PD1 agents release the PD1-PDL1 interaction between CD8 T cells and macrophages to promote cytotoxic antitumor activity