Rapid dissection and model-based optimization of inducible enhancers in human cells using a massively parallel reporter assay

Learning to read and write the transcriptional regulatory code is of central importance to progress in genetic analysis and engineering. Here we describe a massively parallel reporter assay (MPRA) that facilitates the systematic dissection of transcriptional regulatory elements. In MPRA, microarray-synthesized DNA regulatory elements and unique sequence tags are cloned into plasmids to generate a library of reporter constructs. These constructs are transfected into cells and tag expression is assayed by high-throughput sequencing. We apply MPRA to compare >27,000 variants of two inducible enhancers in human cells: a synthetic cAMP-regulated enhancer and the virus-inducible interferon-β enhancer. We first show that the resulting data define accurate maps of functional transcription factor binding sites in both enhancers at single-nucleotide resolution. We then use the data to train quantitative sequence-activity models (QSAMs) of the two enhancers. We show that QSAMs from two cellular states can be combined to design enhancer variants that optimize potentially conflicting objectives, such as maximizing induced activity while minimizing basal activity.National Human Genome Research Institute (U.S.) (grant R01HG004037)National Science Foundation (U.S.) ((NSF) grant PHY-0957573)National Science Foundation (U.S.) (NSF grant PHY-1022140)Broad Institut

QCD and strongly coupled gauge theories : challenges and perspectives

We highlight the progress, current status, and open challenges of QCD-driven physics, in theory and in experiment. We discuss how the strong interaction is intimately connected to a broad sweep of physical problems, in settings ranging from astrophysics and cosmology to strongly coupled, complex systems in particle and condensed-matter physics, as well as to searches for physics beyond the Standard Model. We also discuss how success in describing the strong interaction impacts other fields, and, in turn, how such subjects can impact studies of the strong interaction. In the course of the work we offer a perspective on the many research streams which flow into and out of QCD, as well as a vision for future developments.Peer reviewe

Lung lipid deposition in pneumonias of viral and non-viral aetiology

AbstractPneumonia is an acute respiratory disease of varying aetiology, which drew much attention during the COVID-19 pandemic. Among many thoroughly studied aspects of pneumonia, lipid metabolism has been addressed insufficiently. Here, we report on abnormal lipid metabolism of both COVID-19- and non-COVID-19-associated pneumonias in human lungs. Morphometric analysis revealed extracellular and intracellular lipid depositions, most notably within vessels adjacent to inflamed regions, where they apparently interfere with the blood flow. Lipids were visualized on Sudan III- and Oil Red O-stained cryosections and on OsO4-contrasted semi-thin and ultrathin sections. Chromato-mass spectrometry revealed that unsaturated fatty acid content was elevated at inflammation sites compared with the intact sites of the same lung. The genes involved in lipid metabolism were downregulated in pneumonia, as shown by qPCR andin silicoRNAseq analysis. Thus, pneumonias are associated with marked lipid abnormalities, and therefore lipid metabolism can be considered a target for new therapeutic strategies.</jats:p

Cumulative Effects of Rapid Land-Cover and Land-Use Changes on the Yamal Peninsula, Russia

The Yamal Peninsula in northwest Siberia is undergoing some of the most rapid land-cover and land-use changes in the Arctic due to a combination of gas development, reindeer herding, and climate change. Unusual geological conditions (nutrient-poor sands, massive ground ice and extensive landslides) exacerbate the impacts. These changes will likely increase markedly as transportation corridors are built to transport the gas to market. Understanding the nature, extent, causes and consequences (i.e., the cumulative effects) of the past and ongoing rapid changes on the Yamal is important for effective, long-term decision-making and planning. The cumulative effects to vegetation are the focus of this chapter because the plants are a critical component of the Yamal landscape that support the indigenous Nenets people and their reindeer and also protect the underlying ice-rich permafrost from melting. We are using a combination of ground-based studies (a transect of live locations across the Yamal), remote-sensing studies, and analyses of Nenets land-use activities to develop vegetation-change models that can be used to help anticipate future states of the tundra and how those changes might affect traditional reindeer herding practices and the thermal state of the permafrost. This chapter provides an overview of the approach, some early results, and recommendations for expanding the concept of cumulative-effects analysis to include examining the simultaneous and interactive effects of multiple drivers of change.</p