OBJECTIVES/GOALS: Glioblastoma (GBM) is the most common and aggressive adult primary brain malignancy. Clinically, GBM is refractory to T cell immune checkpoint blockade (ICB), in part due to its dense immune suppressive myeloid stroma. Here we show that myeloid-targeting STING agonists can repolarize the GBM microenvironment to cure ICB-refractory GBM models. METHODS/STUDY POPULATION: Using the synthetic cyclic di-nucleotide STING agonist IACS-8803 (8803) we treated orthotopic ICB-refractory QPP8 orthotopic murine GBM tumors intratumorally. We then analyzed survival and performed high parameter flow cytometry profiling of the tumor immune microenvironment following STING agonist treatment. To assess the contribution of adaptive immunity to STING agonist therapeutic efficacy, we treated orthotopic QPP8 tumors implanted in RAG1 KO mice and monitored survival. RESULTS/ANTICIPATED RESULTS: We found that STING agonist therapy cured murine orthotopic QPP8 tumors, in contrast to ICB that showed no survival benefit. In RAG1-/- mice bearing QPP8 tumors STING agonist therapy extended survival, however, the curative effect observed in wild-type mice was lost in the absence of adaptive immunity. STING agonist-treated QPP8 tumors displayed increased counts of CD8 T cells and NK cells, and decreased CD8 T cell PD1 expression. Infiltration of STING-treated gliomas by Ly6C+ F4/80+ Mono-MDSC substantially increased; however, these cells expressed reduced CD206 and CD163, suggestive of reduced immuno-suppression. Finally, in the cervical LN of QPP8-treated mice the frequency and CD80/CD86 expression of cDC1 cells increased.​ DISCUSSION/SIGNIFICANCE: ICB has failed in GBM, and the suppressive myeloid stroma remains a major barrier to generating anti-GBM T cell responses. Our work shows that STING activation, which primarily targets innate immunity myeloid cells 'upstream' of T cells in the antitumor immunity cycle, can cure ICB-refractory GBM tumors in an adaptive immunity-dependent manner.

OBJECTIVES/GOALS: As hospitals across the nation respond to the need to address community violence, there is a dearth of Hospital-based Violence Intervention Programs (HVIPs) in the South despite having disproportionate rates. This research aims to identify key factors and strategies for implementation of an HVIP among rural patient populations in a southern state. METHODS/STUDY POPULATION: Semi-structured interviews will be conducted with medical providers, social service organizations, and patients transferred from four high-risk rural areas in Arkansas. Data will be analyzed using Framework Analysis, a rapid analysis approach involving framework development, code application, impactful statement identification, and content analysis. Evidence- Based Quality Improvement (EBQI), a group consensus making process, will be conducted to identify key implementation strategies and factors to adapt based interview findings. Priority areas for adaptation will be identified via systematic rating. The EBQI team, including researchers and key rural stakeholders will engage in a series of discussion, vote on final strategies, and develop a guide for future HVIP implementation and pilot testing. RESULTS/ANTICIPATED RESULTS: Findings from this study will result in a prioritized list of barriers and facilitators across sample groups. Factors will be rated by level of importance. Cluster maps will display the relationships among factors. Go and no-go zones will be identified based on importance and feasibility. Implementation strategies will be mapped to barriers and facilitators. DISCUSSION/SIGNIFICANCE: The findings will result in a culturally and geographically relevant HVIP model and package of implementation strategies to test in future hybrid trials (feasibility pilot & multi-site RCT); and shape the future of violence prevention efforts in healthcare settings across the rural South.

Seed retention, and ultimately seed shatter, are extremely important for the efficacy of harvest weed seed control (HWSC) and are likely influenced by various agroecological and environmental factors. Field studies investigated seed-shattering phenology of 22 weed species across three soybean [Glycine max (L.) Merr.]-producing regions in the United States. We further evaluated the potential drivers of seed shatter in terms of weather conditions, growing degree days, and plant biomass. Based on the results, weather conditions had no consistent impact on weed seed shatter. However, there was a positive correlation between individual weed plant biomass and delayed weed seed–shattering rates during harvest. This work demonstrates that HWSC can potentially reduce weed seedbank inputs of plants that have escaped early-season management practices and retained seed through harvest. However, smaller individuals of plants within the same population that shatter seed before harvest pose a risk of escaping early-season management and HWSC.

ABSTRACT IMPACT: This formative evaluation can inform selection and development of implementation strategies for implementing this and other similar interventions in future implementation studies or practice. OBJECTIVES/GOALS: Al-Anon mutual-help groups help concerned others (COs; e.g., families, friends) of persons with an alcohol use disorder better cope with their own problems. Despite widespread availability of Al-Anon meetings, participation is limited. We developed and evaluated an intervention to facilitate CO engagement in Al-Anon. METHODS/STUDY POPULATION: Al-Anon Intensive Referral (AIR) was developed to facilitate COs’ engagement in Al-Anon through four coaching sessions and is being tested in a NIAAA-funded randomized controlled trial (RCT). Consistent with a hybrid type 1 effectiveness-implementation design, we also conducted a formative evaluation to learn about facilitators, barriers and recommendations for AIR implementation in substance use disorder (SUD) treatment programs. We interviewed key informants (director and two staff) at eight sites in the AIR RCT and two ‘naive’ sites unfamiliar with AIR. Sites included community and Veterans Administration (VA) treatment programs in Arkansas, California, and Nebraska. Semi-structured interviews were based on the Consolidated Framework for Implementation Research, and were thematically analyzed. RESULTS/ANTICIPATED RESULTS: Facilitators included AIR’s face validity, adaptability, and alignment with staff values and skills, requiring only minimal training. Several community sites thought AIR would fit with their current practices (e.g. family groups), and some sites reported having sufficient staff available for delivering AIR. Barriers included limited staff time (some sites), and VA sites having limited resources for providing services to COs. Furthermore, many clients have no COs, or COs who are unwilling or unable to engage. Recommendations included fitting AIR within existing workflows and focusing on COs with highest readiness. Participants also thought AIR could be adapted as an online or smartphone app, which may expand its reach to younger and more tech-savvy populations while decreasing staff burden. DISCUSSION/SIGNIFICANCE OF FINDINGS: AIR has strong potential for implementation, but sites vary on implementation capacity and readiness. Most sites could implement it partially (e.g., case-by-case basis), and sites with sufficient capacity (e.g., family groups, staff time) could implement it more fully. An app-based AIR could help mitigate some barriers.

Potential effectiveness of harvest weed seed control (HWSC) systems depends upon seed shatter of the target weed species at crop maturity, enabling its collection and processing at crop harvest. However, seed retention likely is influenced by agroecological and environmental factors. In 2016 and 2017, we assessed seed-shatter phenology in 13 economically important broadleaf weed species in soybean [Glycine max (L.) Merr.] from crop physiological maturity to 4 wk after physiological maturity at multiple sites spread across 14 states in the southern, northern, and mid-Atlantic United States. Greater proportions of seeds were retained by weeds in southern latitudes and shatter rate increased at northern latitudes. Amaranthus spp. seed shatter was low (0% to 2%), whereas shatter varied widely in common ragweed (Ambrosia artemisiifolia L.) (2% to 90%) over the weeks following soybean physiological maturity. Overall, the broadleaf species studied shattered less than 10% of their seeds by soybean harvest. Our results suggest that some of the broadleaf species with greater seed retention rates in the weeks following soybean physiological maturity may be good candidates for HWSC.

Seed shatter is an important weediness trait on which the efficacy of harvest weed seed control (HWSC) depends. The level of seed shatter in a species is likely influenced by agroecological and environmental factors. In 2016 and 2017, we assessed seed shatter of eight economically important grass weed species in soybean [Glycine max (L.) Merr.] from crop physiological maturity to 4 wk after maturity at multiple sites spread across 11 states in the southern, northern, and mid-Atlantic United States. From soybean maturity to 4 wk after maturity, cumulative percent seed shatter was lowest in the southern U.S. regions and increased moving north through the states. At soybean maturity, the percent of seed shatter ranged from 1% to 70%. That range had shifted to 5% to 100% (mean: 42%) by 25 d after soybean maturity. There were considerable differences in seed-shatter onset and rate of progression between sites and years in some species that could impact their susceptibility to HWSC. Our results suggest that many summer annual grass species are likely not ideal candidates for HWSC, although HWSC could substantially reduce their seed output during certain years.

Across the world, in many places in which English is not widely spoken, English text often appears on posters, storefronts, billboards, street signs, warning signs, menus, and many other forms of publicly visible written texts. English is often featured alongside one or more additional languages. These signs are typically seen as unremarkable by passersby looking to buy goods, for information regarding prohibited activity, or even just for a comfortable coffee shop. An increasingly robust scholarly literature has examined these signs since the publication of Landry and Bourhis' (1997) foundational study which popularized the term linguistic landscape. Linguistic landscape scholarship now includes large-scale quantitative examinations of publicly visible signs in cities such as Tokyo (Backhaus, 2007), smaller-scale examinations of the written language within individual workplaces in a single building (Hanauer, 2010), studies on mobile linguistic landscapes such as shopping bags (Alomoush, 2019) and many other studies conducted in widely varying locations with widely varying foci (McKiernan, 2019; Yuan, 2019).

We propose the nasal administration of calcium-enriched physiological salts as a new hygienic intervention with possible therapeutic application as a response to the rapid and tenacious spread of COVID-19. We test the effectiveness of these salts against viral and bacterial pathogens in animals and humans. We find that aerosol administration of these salts to the airways diminishes the exhalation of the small particles that face masks fail to filter and, in the case of an influenza swine model, completely block airborne transmission of disease. In a study of 10 human volunteers (5 less than 65 years and 5 older than 65 years), we show that delivery of a nasal saline comprising calcium and sodium salts quickly (within 15 min) and durably (up to at least 6 h) diminishes exhaled particles from the human airways. Being predominantly smaller than 1 μm, these particles are below the size effectively filtered by conventional masks. The suppression of exhaled droplets by the nasal delivery of calcium-rich saline with aerosol droplet size of around 10 μm suggests the upper airways as a primary source of bioaerosol generation. The suppression effect is especially pronounced (99%) among those who exhale large numbers of particles. In our study, we found this high-particle exhalation group to correlate with advanced age. We argue for a new hygienic practice of nasal cleansing by a calcium-rich saline aerosol, to complement the washing of hands with ordinary soap, use of a face mask, and social distancing.

Seven half-day regional listening sessions were held between December 2016 and April 2017 with groups of diverse stakeholders on the issues and potential solutions for herbicide-resistance management. The objective of the listening sessions was to connect with stakeholders and hear their challenges and recommendations for addressing herbicide resistance. The coordinating team hired Strategic Conservation Solutions, LLC, to facilitate all the sessions. They and the coordinating team used in-person meetings, teleconferences, and email to communicate and coordinate the activities leading up to each regional listening session. The agenda was the same across all sessions and included small-group discussions followed by reporting to the full group for discussion. The planning process was the same across all the sessions, although the selection of venue, time of day, and stakeholder participants differed to accommodate the differences among regions. The listening-session format required a great deal of work and flexibility on the part of the coordinating team and regional coordinators. Overall, the participant evaluations from the sessions were positive, with participants expressing appreciation that they were asked for their thoughts on the subject of herbicide resistance. This paper details the methods and processes used to conduct these regional listening sessions and provides an assessment of the strengths and limitations of those processes.

Herbicide resistance is ‘wicked’ in nature; therefore, results of the many educational efforts to encourage diversification of weed control practices in the United States have been mixed. It is clear that we do not sufficiently understand the totality of the grassroots obstacles, concerns, challenges, and specific solutions needed for varied crop production systems. Weed management issues and solutions vary with such variables as management styles, regions, cropping systems, and available or affordable technologies. Therefore, to help the weed science community better understand the needs and ideas of those directly dealing with herbicide resistance, seven half-day regional listening sessions were held across the United States between December 2016 and April 2017 with groups of diverse stakeholders on the issues and potential solutions for herbicide resistance management. The major goals of the sessions were to gain an understanding of stakeholders and their goals and concerns related to herbicide resistance management, to become familiar with regional differences, and to identify decision maker needs to address herbicide resistance. The messages shared by listening-session participants could be summarized by six themes: we need new herbicides; there is no need for more regulation; there is a need for more education, especially for others who were not present; diversity is hard; the agricultural economy makes it difficult to make changes; and we are aware of herbicide resistance but are managing it. The authors concluded that more work is needed to bring a community-wide, interdisciplinary approach to understanding the complexity of managing weeds within the context of the whole farm operation and for communicating the need to address herbicide resistance.

OBJECTIVES/SPECIFIC AIMS: The primary objective of this effort is to develop and distribute an easy to use i2b2 component that is capable of evaluating diverse complex relationships for a wide variety of exposures and outcomes over time. In this manner we are able to leverage the unique design of the i2b2 database to support health services research, comparative effectiveness, and quality improvement using a single tool. Furthermore, our novel database redesign has the potential to provide user-friendly access to individual and group CHC data for CER. METHODS/STUDY POPULATION: For this project we used software experts, clinical informatics specialists, and the existing i2b2 open-source software to convert our legacy HOME Cell into a web-client version. The tool will be used to study health outcomes within a network of Boston based Community Health Centers and the largest safety-net hospital in New England, Boston Medical Center. RESULTS/ANTICIPATED RESULTS: The new web-client HOME Cell will allow i2b2 users to model virtually any exposure (including therapeutic interventions such as medications or tests) in i2b2 against any outcome accounting for complex temporal relationships and other factors. In addition we plan to use our new Community Health Center views to enhance our community engagement activities by allowing direct access to their data for our partners. DISCUSSION/SIGNIFICANCE OF IMPACT: Our project addresses multiple national priorities related to data sharing, clinical research informatics, and comparative effectiveness. The web-client version of the HOME Cell substantially improves our community’s access to HOME Cell functionality and is a novel, sharable resource for use within the CTSA/NCATS community. Our approach provides a new way to perform large-scale collaborative research without the need to actually move patient-level data and has demonstrated that CER, health services research, and quality measurement can share a common framework. In addition, and as demonstrated in our earlier pilot work, the HOME Cell also has the potential to support large-scale multivariate analyses in a distributed manner that does not require sharing of patient-level data. We believe our approach has great promise for supporting the reuse of clinical data for rapid, transparent, health outcome assessments on a national scale. Our efforts support multiple strategic goals including: (1) support for building national clinical and translational research capacity by enhancing a broadly adopted informatics tool (i2b2); (2) enhanced consortium-wide collaborations by offering a tool that can be easily shared within the CTSA network to support multi-institutional collaboration; and (3) improving the health of our communities by offering a tool that has the potential to provide new insights into health care processes and outcomes that could drive innovation and improvement activities.

A postemergence (POST) timing study was conducted on established populations of burcucumber (Sicyos angulatus) in corn (Zea mays), and a second study examined the residual activity of several herbicides for burcucumber control under greenhouse conditions. In the field study, flumiclorac, halosulfuron, primisulfuron, CGA 152005, and CGA 152005 + primisulfuron (45, 71, 40, 40, and 20 + 20 g ai/ha, respectively) were applied at two POST timings. CGA 152005, primisulfuron, and the combination provided greater than 85% control of burcucumber 14 wk after planting (WAP). Flumiclorac and halosulfuron provided 60% control or less by 8 WAP. Timing of the POST applications did not influence burcucumber control by 11 WAP with any herbicide. In the greenhouse, germinated burcucumber seeds were placed in soil treated with atrazine, chlorimuron, primisulfuron, or CGA 152005 at normal field use rates. All treatments provided similar residual control early; however by 4 wk after treatment (WAT), control from atrazine was less than 10% compared to 69% for chlorimuron and about 50% for primisulfuron and CGA 152005. This research suggests that CGA 152005 and primisulfuron can both be effective for managing burcucumber in corn, whereas flumiclorac and halosulfuron proved ineffective.

An experiment was conducted in 1998 and 1999 in southeastern Pennsylvania to examine the effect of tillage and soybean row spacing on burcucumber emergence and growth. A second experiment evaluated postemergence (POST) soybean herbicides on burcucumber control. In the tillage and row spacing study, a glyphosate-resistant soybean variety was planted in no-till and reduced-tillage systems in 38- and 76-cm row spacings. In the POST herbicide experiment, chlorimuron, glyphosate, CGA-277476, thifensulfuron, and several combinations of these herbicides were applied at two different POST application timings in 38-cm row soybean planted in a reduced-tillage system. In the tillage and row spacing study, burcucumber emergence was greatest starting in late May through mid-June and mostly ceased by early July, regardless of tillage system or row spacing. Although there was no difference in germination period in either tillage system, preplant tillage increased the number of emerged plants by 110% in 1997 and 70% in 1998 compared to the no-till system. Row spacing had no effect on burcucumber emergence or biomass production. In general, most POST herbicide programs controlled burcucumber, and there was no difference between early POST and mid-POST application timings. Chlorimuron at 13 g ai/ha, chlorimuron plus thifensulfuron, glyphosate, glyphosate plus chlorimuron, and glyphosate plus CGA-277476 provided 87% or greater control of burcucumber 12 wk after planting. These herbicides reduced burcucumber density and biomass by more than 56% in 1997 and 96% in 1998.

Experiments examining burcucumber management in glufosinate-resistant (GR) and imidazolinone-resistant (IMI) corn were conducted in 1997 and 1998 in southeastern Pennsylvania. GR corn was planted in 38- and 76-cm rows, and postemergence (POST) treatments of glufosinate and glufosinate plus atrazine were applied to corn at the V4 or V5 growth stage. In a second study, IMI corn was planted in 76-cm rows, and 15 preemergence (PRE) and POST herbicide programs were evaluated. Herbicide treatments included RPA-201772, CGA 152005, simazine, imazethapyr plus imazapyr, imazamox, chlorimuron plus thifensulfuron, nicosulfuron plus rimsulfuron plus atrazine, CGA 152005 plus primisulfuron, and combinations with atrazine. Burcucumber germinated throughout the growing season, with greatest emergence occurring in early June, gradually decreasing to minimal emergence by mid-July. Glufosinate alone controlled burcucumber 79 to 90% 7 weeks after planting (WAP) regardless of application timing or row spacing. By 10 to 13 WAP, control was 82% or less due to lack of residual control and new burcucumber emergence. Row spacing had little effect on burcucumber emergence or control and appears to have little impact on burcucumber management in corn. In general, PRE herbicide programs were less effective than POST programs, although PRE treatments containing atrazine equaled some POST herbicides. POST-applied chlorimuron plus thifensulfuron, nicosulfuron plus rimsulfuron plus atrazine, and CGA 152005 plus primisulfuron controlled burcucumber greater than 80 and 90% in 1997 and 1998, respectively. Imazethapyr plus imazapyr and imazamox applied POST controlled burcucumber 66% 10 WAP. Adding atrazine to POST herbicide programs did not increase control, with the exception of imazethapyr plus imazapyr.

An experiment examining the effect of burcucumber emergence date and corn competition on burcucumber fecundity was conducted in 1997 and 1998 in central Pennsylvania. Burcucumber seedlings were transplanted in the field in approximately 10-d intervals starting in late May through mid-August with or without competition from corn. Burcucumber plants grown without competition from corn produced 716 g dry matter and 4,500 seeds plant−1 in 1997 and 607 g dry matter and 1,800 seeds plant−1 in 1998. Biomass was greatest for plants established in late May, whereas seed production was greatest for plants established in mid-June. Although seed numbers were reduced in comparison to the May and June establishment periods, plants established as late as August still produced seed. Burcucumber established in corn produced 96% less dry matter and seed than the plants in a noncompetitive environment in both years of the study. Although the growth and seed production of burcucumber grown in corn was drastically reduced, plants established in mid-July still produced seed.