GRK6 regulates the hemostatic response to injury through its rate-limiting effects on GPCR signaling in platelets.

G protein-coupled receptors (GPCRs) mediate the majority of platelet activation in response to agonists. However, questions remain regarding the mechanisms that provide negative feedback toward activated GPCRs to limit platelet activation and thrombus formation. Here we provide the first evidence that GPCR kinase 6 (GRK6) serves this role in platelets, using GRK6-/- mice generated by CRISPR-Cas9 genome editing to examine the consequences of GRK6 knockout on GPCR-dependent signaling. Hemostatic thrombi formed in GRK6-/- mice are larger than in wild-type (WT) controls during the early stages of thrombus formation, with a rapid increase in platelet accumulation at the site of injury. GRK6-/- platelets have increased platelet activation, but in an agonist-selective manner. Responses to PAR4 agonist or adenosine 5\u27-diphosphate stimulation in GRK6-/- platelets are increased compared with WT littermates, whereas the response to thromboxane A2 (TxA2) is normal. Underlying these changes in GRK6-/- platelets is an increase in Ca2+ mobilization, Akt activation, and granule secretion. Furthermore, deletion of GRK6 in human MEG-01 cells causes an increase in Ca2+ response and PAR1 surface expression in response to thrombin. Finally, we show that human platelet activation in response to thrombin causes an increase in binding of GRK6 to PAR1, as well as an increase in the phosphorylation of PAR1. Deletion of GRK6 in MEG-01 cells causes a decrease in PAR1 phosphorylation. Taken together, these data show that GRK6 regulates the hemostatic response to injury through PAR- and P2Y12-mediated effects, helping to limit the rate of platelet activation during thrombus growth and prevent inappropriate platelet activation

TC21/RRas2 regulates glycoprotein VI–FcRγ‐mediated platelet activation and thrombus stability

Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/145353/1/jth14197.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/145353/2/jth14197_am.pd

Comparative transcriptomics of pathogenic and non-pathogenic Listeria species

Comparative RNA-seq analysis of two related pathogenic and non-pathogenic bacterial strains reveals a hidden layer of divergence in the non-coding genome as well as conserved, widespread regulatory structures called ‘Excludons', which mediate regulation through long non-coding antisense RNAs

Data report: composite depth scale and splice revision for IODP Site U1488 (Expedition 363 Western Pacific Warm Pool) using XRF core scanning data and core images

The Western Pacific Warm Pool (WPWP) is a major source of heat and moisture to the atmosphere. Small perturbations in WPWP sea-surface temperatures greatly influence local Hadley and Walker cells, thereby affecting global atmospheric circulation patterns. International Ocean Discovery Program (IODP) Expedition 363 sought to document the regional expression and driving mechanisms of WPWP climate variability during the Neogene on millennial, orbital, and geological timescales. Located in the heart of the WPWP, IODP Site U1488 (02°02.59ʹN, 141°45.29ʹE) was drilled in 2604 m water depth on the southern part of the Eauripik Rise in the Caroline Basin. At Site U1488, a continuous shipboard composite stratigraphic section from 0 to ~331 m core composite depth below seafloor (CCSF) was compiled using high-resolution shipboard physical property data from three holes. This section comprises upper Miocene to recent foraminifer-rich nannofossil ooze and foraminifer-nannofossil ooze, making Site U1488 ideally suited to reconstruct the paleoceanographic history of the central WPWP region. However, the high carbonate content (>90% below ~180 m CCSF) of Site U1488 sediments means that the physical property data sets commonly used for splice construction (gamma ray attenuation bulk density, magnetic susceptibility, and natural gamma radiation) were too low amplitude to provide robust constraints on splice tie points below 120 m CCSF. As a result, P-wave data, which are relatively untested as a correlation tool, became critical for correlating between holes. Here, we verify and extend the Site U1488 shipboard composite splice using high-resolution (2 cm) X-ray fluorescence Ba/Sr core scanning data combined with composite linescan images. Overall, using these data at Site U1488 resulted in revised core offsets that differ by up to 0.84 m relative to the shipboard core offsets and a composite depth scale down to 329.33 m revised CCSF. The revised splice will allow optimization of postexpedition research and ensure that high-resolution studies of Site U1488 are conducted on a continuous stratigraphic section

Lycopene Cyclase Paralog CruP Protects Against Reactive Oxygen Species in Oxygenic Photosynthetic Organisms

In photosynthetic organisms, carotenoids serve essential roles in photosynthesis and photoprotection. A previous report designated CruP as a secondary lycopene cyclase involved in carotenoid biosynthesis [Maresca J, et al. (2007) Proc Natl Acad Sci USA 104:11784–11789]. However, we found that cruP KO or cruP overexpression plants do not exhibit correspondingly reduced or increased production of cyclized carotenoids, which would be expected if CruP was a lycopene cyclase. Instead, we show that CruP aids in preventing accumulation of reactive oxygen species (ROS), thereby reducing accumulation of β-carotene-5,6-epoxide, a ROS-catalyzed autoxidation product, and inhibiting accumulation of anthocyanins, which are known chemical indicators of ROS. Plants with a nonfunctional cruP accumulate substantially higher levels of ROS and β-carotene-5,6-epoxide in green tissues. Plants overexpressing cruP show reduced levels of ROS, β-carotene-5,6-epoxide, and anthocyanins. The observed up-regulation of cruP transcripts under photoinhibitory and lipid peroxidation-inducing conditions, such as high light stress, cold stress, anoxia, and low levels of CO2, fits with a role for CruP in mitigating the effects of ROS. Phylogenetic distribution of CruP in prokaryotes showed that the gene is only present in cyanobacteria that live in habitats characterized by large variation in temperature and inorganic carbon availability. Therefore, CruP represents a unique target for developing resilient plants and algae needed to supply food and biofuels in the face of global climate change

Connections of climate change and variability to large and extreme forest fires in southeast Australia

The 2019/20 Black Summer bushfire disaster in southeast Australia was unprecedented: the extensive area of forest burnt, the radiative power of the fires, and the extraordinary number of fires that developed into extreme pyroconvective events were all unmatched in the historical record. Australia’s hottest and driest year on record, 2019, was characterised by exceptionally dry fuel loads that primed the landscape to burn when exposed to dangerous fire weather and ignition. The combination of climate variability and long-term climate trends generated the climate extremes experienced in 2019, and the compounding effects of two or more modes of climate variability in their fire-promoting phases (as occurred in 2019) has historically increased the chances of large forest fires occurring in southeast Australia. Palaeoclimate evidence also demonstrates that fire-promoting phases of tropical Pacific and Indian ocean variability are now unusually frequent compared with natural variability in preindustrial times. Indicators of forest fire danger in southeast Australia have already emerged outside of the range of historical experience, suggesting that projections made more than a decade ago that increases in climate-driven fire risk would be detectable by 2020, have indeed eventuated. The multiple climate change contributors to fire risk in southeast Australia, as well as the observed non-linear escalation of fire extent and intensity, raise the likelihood that fire events may continue to rapidly intensify in the future. Improving local and national adaptation measures while also pursuing ambitious global climate change mitigation efforts would provide the best strategy for limiting further increases in fire risk in southeast Australia

Engineering key components in a synthetic eukaryotic signal transduction pathway

Signal transduction underlies how living organisms detect and respond to stimuli. A goal of synthetic biology is to rewire natural signal transduction systems. Bacteria, yeast, and plants sense environmental aspects through conserved histidine kinase (HK) signal transduction systems. HK protein components are typically comprised of multiple, relatively modular, and conserved domains. Phosphate transfer between these components may exhibit considerable cross talk between the otherwise apparently linear pathways, thereby establishing networks that integrate multiple signals. We show that sequence conservation and cross talk can extend across kingdoms and can be exploited to produce a synthetic plant signal transduction system. In response to HK cross talk, heterologously expressed bacterial response regulators, PhoB and OmpR, translocate to the nucleus on HK activation. Using this discovery, combined with modification of PhoB (PhoB-VP64), we produced a key component of a eukaryotic synthetic signal transduction pathway. In response to exogenous cytokinin, PhoB-VP64 translocates to the nucleus, binds a synthetic PlantPho promoter, and activates gene expression. These results show that conserved-signaling components can be used across kingdoms and adapted to produce synthetic eukaryotic signal transduction pathways

Mutation Detection with Next-Generation Resequencing through a Mediator Genome

The affordability of next generation sequencing (NGS) is transforming the field of mutation analysis in bacteria. The genetic basis for phenotype alteration can be identified directly by sequencing the entire genome of the mutant and comparing it to the wild-type (WT) genome, thus identifying acquired mutations. A major limitation for this approach is the need for an a-priori sequenced reference genome for the WT organism, as the short reads of most current NGS approaches usually prohibit de-novo genome assembly. To overcome this limitation we propose a general framework that utilizes the genome of relative organisms as mediators for comparing WT and mutant bacteria. Under this framework, both mutant and WT genomes are sequenced with NGS, and the short sequencing reads are mapped to the mediator genome. Variations between the mutant and the mediator that recur in the WT are ignored, thus pinpointing the differences between the mutant and the WT. To validate this approach we sequenced the genome of Bdellovibrio bacteriovorus 109J, an obligatory bacterial predator, and its prey-independent mutant, and compared both to the mediator species Bdellovibrio bacteriovorus HD100. Although the mutant and the mediator sequences differed in more than 28,000 nucleotide positions, our approach enabled pinpointing the single causative mutation. Experimental validation in 53 additional mutants further established the implicated gene. Our approach extends the applicability of NGS-based mutant analyses beyond the domain of available reference genomes

Making the cut: The production of 'self-harm' in post-1945 Anglo-Saxon psychiatry.

'Deliberate self-harm', 'self-mutilation' and 'self-injury' are just some of the terms used to describe one of the most prominent issues in British mental health policy in recent years. This article demonstrates that contemporary literature on 'self-harm' produces this phenomenon (to varying extents) around two key characteristics. First, this behaviour is predominantly performed by those identified as female. Second, this behaviour primarily involves cutting the skin. These constitutive characteristics are traced back to a corpus of literature produced in the 1960s and 1970s in North American psychiatric inpatient institutions; analysis shows how pre-1960 works were substantially different. Finally, these gendered and behavioural assertions are shown to be the result of historically specific processes of exclusion and emphasis