OBJECTIVES/GOALS: Glioblastomas (GBMs) are heterogeneous, treatment-resistant tumors that are driven by populations of cancer stem cells (CSCs). In this study, we perform an epigenetic-focused functional genomics screen in GBM organoids and identify WDR5 as an essential epigenetic regulator in the SOX2-enriched, therapy resistant cancer stem cell niche. METHODS/STUDY POPULATION: Despite their importance for tumor growth, few molecular mechanisms critical for CSC population maintenance have been exploited for therapeutic development. We developed a spatially resolved loss-of-function screen in GBM patient-derived organoids to identify essential epigenetic regulators in the SOX2-enriched, therapy resistant niche. Our niche-specific screens identified WDR5, an H3K4 histone methyltransferase responsible for activating specific gene expression, as indispensable for GBM CSC growth and survival. RESULTS/ANTICIPATED RESULTS: In GBM CSC models, WDR5 inhibitors blocked WRAD complex assembly and reduced H3K4 trimethylation and expression of genes involved in CSC-relevant oncogenic pathways. H3K4me3 peaks lost with WDR5 inhibitor treatment occurred disproportionally on POU transcription factor motifs, required for stem cell maintenance and including the POU5F1(OCT4)::SOX2 motif. We incorporated a SOX2/OCT4 motif driven GFP reporter system into our CSC cell models and found that WDR5 inhibitor treatment resulted in dose-dependent silencing of stem cell reporter activity. Further, WDR5 inhibitor treatment altered the stem cell state, disrupting CSC in vitro growth and self-renewal as well as in vivo tumor growth. DISCUSSION/SIGNIFICANCE: Our results unveiled the role of WDR5 in maintaining the CSC state in GBM and provide a rationale for therapeutic development of WDR5 inhibitors for GBM and other advanced cancers. This conceptual and experimental framework can be applied to many cancers, and can unmask unique microenvironmental biology and rationally designed combination therapies.

After five positive randomized controlled trials showed benefit of mechanical thrombectomy in the management of acute ischemic stroke with emergent large-vessel occlusion, a multi-society meeting was organized during the 17th Congress of the World Federation of Interventional and Therapeutic Neuroradiology in October 2017 in Budapest, Hungary. This multi-society meeting was dedicated to establish standards of practice in acute ischemic stroke intervention aiming for a consensus on the minimum requirements for centers providing such treatment. In an ideal situation, all patients would be treated at a center offering a full spectrum of neuroendovascular care (a level 1 center). However, for geographical reasons, some patients are unable to reach such a center in a reasonable period of time. With this in mind, the group paid special attention to define recommendations on the prerequisites of organizing stroke centers providing medical thrombectomy for acute ischemic stroke, but not for other neurovascular diseases (level 2 centers). Finally, some centers will have a stroke unit and offer intravenous thrombolysis, but not any endovascular stroke therapy (level 3 centers). Together, these level 1, 2, and 3 centers form a complete stroke system of care. The multi-society group provides recommendations and a framework for the development of medical thrombectomy services worldwide.

We used pollen and high-resolution charcoal analysis of lake sediment to reconstruct a 7600 yr vegetation and fire history from Anthony Lake, located in the Blue Mountains of northeastern Oregon. From 7300 to 6300 cal yr BP, the forest was composed primarily of Populus, and fire was common, indicating warm, dry conditions. From 6300 to 3000 cal yr BP, Populus declined as Pinus and Picea increased in abundance and fire became less frequent, suggesting a shift to cooler, wetter conditions. From 3000 cal yr BP to present, modern-day forests composed of Pinus and Abies developed, and from 1650 cal yr BP to present, fires increased. We utilized the modern climate-analogue approach to explain the potential synoptic climatological processes associated with regional fire. The results indicate that years with high fire occurrence experience positive 500 mb height anomalies centered over the Great Basin, with anomalous southerly component of flow delivering dry air into the region and with associated sinking motions to further suppress precipitation. It is possible that such conditions became more common over the last 1650 cal yr BP, supporting an increase in fire despite the shift to more mesic conditions.

The phenomenon of buying-shopping disorder (BSD) was described over 100 years ago. Definitions of BSD refer to extreme preoccupation with shopping and buying, to impulses to purchase that are experienced as irresistible, and to recurrent maladaptive buying excesses that lead to distress and impairments. Efforts to stop BSD episodes are unsuccessful, despite the awareness of repeated break-downs in self-regulation, experiences of post-purchase guilt and regret, comorbid psychiatric disorders, reduced quality of life, familial discord, work impairment, financial problems, and other negative consequences. A recent meta-analysis indicated an estimated point prevalence of BSD of 5%. In this narrative review, the authors offer a perspective to consider BSD as a mental health condition and to classify this disorder as a behavioral addiction, based on both research data and on long-standing clinical experience.

Business and public partnerships in socially responsible behavior have become a central pillar of global governance, but one that is unevenly developed in different countries. Despite the transnational character of business operations, national context is of theoretical as well as policy significance. To explain cross-country variation in corporate commitment to social responsibility we investigate the political conditions that encourage firms to participate in the United Nations Global Compact. Drawing on a theory of corporate social responsibility as motivated by self-interest and external pressure, we examine the influence of external actors and the locally specific mobilization of bias. Analyzing participation levels in 145 countries, we find that a democratic regime and Global Compact participation by countervailing groups are associated with higher levels of business participation in the program. Contrary to earlier studies relying on smaller numbers of countries, we find no evidence that a country's relationship with the UN or the domestic political strength of environmental interests account for cross-national variation in corporate engagement with the Global Compact.

High-resolution charcoal and pollen analyses were used to reconstruct a 12,000-yr-long fire and vegetation history of the Tumalo Lake watershed and to examine the short-term effects that tephra deposition have on forest composition and fire regime. The record suggests that, from 12,000 to 9200 cal yr BP, the watershed was dominated by an open Pinus forest with Artemisia as a common understory species. Fire episodes occurred on average every 115 yr. Beginning around 9200 cal yr BP, and continuing to the present, Abies became more common while Artemisia declined, suggesting the development of a closed forest structure and a decrease in the frequency of fire episodes, occurring on average every 160 yr. High-resolution pollen analyses before and after the emplacement of three distinct tephra deposits in the watershed suggest that nonarboreal species were most affected by tephra events and that recovery of the vegetation community to previous conditions took between 40 and 100 yr. Changes in forest composition were not associated with tephra depositional events or changes in fire-episode frequency, implying that the regional climate is the more important control on long-term forest composition and structure of the vegetation in the Cascade Range.

Woody plant herbicide screening techniques were evaluated to expedite the screening process and decrease amounts of herbicide active ingredient required. Rapid greenhouse screening of woody plant seedlings was performed in less than 6 months, and rapid seed screening was performed in less than 20 days. A traditional field screen, requiring 10 months from application to final evaluation, was performed for comparison and regression modeling purposes. Imazapyr and triclopyr were used as test chemicals and linear regressions were generated to predict traditional field screen results from rapid screens. Significant regressions were produced that predicted field responses of loblolly pine, sweetgum, and yellow-poplar with the use of both herbicides and either rapid screening technique. This indicated that rapid screening techniques could determine herbicide efficacy and/or species spectrum in much less time with significantly less herbicide. Rapid greenhouse screens of triclopyr produced more statistically significant regressions than those using imazapyr. Rapid seed screens could estimate species spectrum within 5 days after treatment. These results indicate that rapid greenhouse screen and rapid seed screen techniques can provide woody plant herbicide developers initial efficacy and spectrum of control data in a cost- and-time effective manner.

The selection of herbicide-resistant weeds in grain sorghum production has prompted researchers to explore alternative herbicides to prevent, delay, and manage herbicide-resistant weed biotypes. Greenhouse and field experiments were conducted to evaluate the differential response of sorghum hybrids to POST application of mesotrione. In a greenhouse experiment, 85 sorghum hybrids were treated with 0, 52, 105, 210, and 315 g ai/ha mesotrione when plants were at the three- to four-leaf collar stage. Sorghum response ranged from susceptible to tolerant sorghum hybrids. ‘Pioneer 84G62’, ‘Pioneer 85G01’, and ‘Triumph TR 438’ were the three most susceptible, whereas ‘Dekalb DKS35-70’, ‘Frontier F222E’, and ‘Asgrow Seneca’ were the three most tolerant hybrids. One week after treatment (WAT), the mesotrione rate causing 50% visible injury ranged from 121 to 184 and 64 to 91 g/ha in the most tolerant and susceptible hybrids, respectively. Mesotrione dose–response studies were conducted under field conditions on four sorghum hybrids. One WAT, injury symptoms were greater (up to 23%) in Pioneer 85G01 than in Asgrow Seneca (< 14%). However, all plants appeared normal by the end of the growing season. In addition, sorghum yields were not reduced by mesotrione treatments as verified by correlation coefficient analysis.

Although acknowledged to be variable and subjective, manual annotation of cryo-electron tomography data is commonly used to answer structural questions and to create a “ground truth” for evaluation of automated segmentation algorithms. Validation of such annotation is lacking, but is critical for understanding the reproducibility of manual annotations. Here, we used voxel-based similarity scores for a variety of specimens, ranging in complexity and segmented by several annotators, to quantify the variation among their annotations. In addition, we have identified procedures for merging annotations to reduce variability, thereby increasing the reliability of manual annotation. Based on our analyses, we find that it is necessary to combine multiple manual annotations to increase the confidence level for answering structural questions. We also make recommendations to guide algorithm development for automated annotation of features of interest.