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.

Research among non-industrial societies suggests that body kinematics adopted during running vary between groups according to the cultural importance of running. Among groups in which running is common and an important part of cultural identity, runners tend to adopt what exercise scientists and coaches consider to be good technique for avoiding injury and maximising performance. In contrast, among groups in which running is not particularly culturally important, people tend to adopt suboptimal technique. This paper begins by describing key elements of good running technique, including landing with a forefoot or midfoot strike pattern and leg oriented roughly vertically. Next, we review evidence from non-industrial societies that cultural attitudes about running associate with variation in running techniques. Then, we present new data from Tsimane forager–horticulturalists in Bolivia. Our findings suggest that running is neither a common activity among the Tsimane nor is it considered an important part of cultural identity. We also demonstrate that when Tsimane do run, they tend to use suboptimal technique, specifically landing with a rearfoot strike pattern and leg protracted ahead of the knee (called overstriding). Finally, we discuss processes by which culture might influence variation in running techniques among non-industrial societies, including self-optimisation and social learning.

In the USA, as many as 20 % of recruits sustain stress fractures during basic training. In addition, approximately one-third of female recruits develop Fe deficiency upon completion of training. Fe is a cofactor in bone collagen formation and vitamin D activation, thus we hypothesised Fe deficiency may be contributing to altered bone microarchitecture and mechanics during 12-weeks of increased mechanical loading. Three-week old female Sprague Dawley rats were assigned to one of four groups: Fe-adequate sedentary, Fe-deficient sedentary, Fe-adequate exercise and Fe-deficient exercise. Exercise consisted of high-intensity treadmill running (54 min 3×/week). After 12-weeks, serum bone turnover markers, femoral geometry and microarchitecture, mechanical properties and fracture toughness and tibiae mineral composition and morphometry were measured. Fe deficiency increased the bone resorption markers C-terminal telopeptide type I collagen and tartate-resistant acid phosphatase 5b (TRAcP 5b). In exercised rats, Fe deficiency further increased bone TRAcP 5b, while in Fe-adequate rats, exercise increased the bone formation marker procollagen type I N-terminal propeptide. In the femur, exercise increased cortical thickness and maximum load. In the tibia, Fe deficiency increased the rate of bone formation, mineral apposition and Zn content. These data show that the femur and tibia structure and mechanical properties are not negatively impacted by Fe deficiency despite a decrease in tibiae Fe content and increase in serum bone resorption markers during 12-weeks of high-intensity running in young growing female rats.

Community research advisory councils (C-RAC) bring together community members with interest in research to support design, evaluation, and dissemination of research in the communities they represent. There are few ways for early career researchers, such as TL1 trainees, to develop skills in community-engaged research, and there are limited opportunities for C-RAC members to influence early career researchers. In our novel training collaboration, TL1 trainees presented their research projects to C-RAC members who provided feedback. We present on initial evidence of student learning and summarize lessons learned that TL1 programs and C-RACs can incorporate into future collaborations.

Sub-acute ruminal acidosis (SARA) can reduce the production efficiency and impair the welfare of cattle, potentially in all production systems. The aim of this study was to characterise measurable postmortem observations from divergently managed intensive beef finishing farms with high rates of concentrate feeding. At the time of slaughter, we obtained samples from 19 to 20 animals on each of 6 beef finishing units (119 animals in total) with diverse feeding practices, which had been subjectively classified as being high risk (three farms) or low risk (three farms) for SARA on the basis of the proportions of barley, silage and straw in the ration. We measured the concentrations of histamine, lipopolysaccharide (LPS), lactate and other short-chain fatty acids (SCFAs) in ruminal fluid, LPS and SCFA in caecal fluid. We also took samples of the ventral blind sac of the rumen for histopathology, immunohistopathology and gene expression. Subjective assessments were made of the presence of lesions on the ruminal wall, the colour of the lining of the ruminal wall and the shape of the ruminal papillae. Almost all variables differed significantly and substantially among farms. Very few pathological changes were detected in any of the rumens examined. The animals on the high-risk diets had lower concentrations of SCFA and higher concentrations of lactate and LPS in the ruminal fluid. Higher LPS concentrations were found in the caecum than the rumen but were not related to the risk status of the farm. The diameters of the stratum granulosum, stratum corneum and of the vasculature of the papillae, and the expression of the gene TLR4 in the ruminal epithelium were all increased on the high-risk farms. The expression of IFN-γ and IL-1β and the counts of cluster of differentiation 3 positive and major histocompatibility complex class two positive cells were lower on the high-risk farms. High among-farm variation and the unbalanced design inherent in this type of study in the field prevented confident assignment of variation in the dependent variables to individual dietary components; however, the CP percentage of the total mixed ration DM was the factor that was most consistently associated with the variables of interest. Despite the strong effect of farm on the measured variables, there was wide inter-animal variation.

Corrugated mats of palygorskite occurring in regolith overlying a fissured limestone were examined by X-ray diffraction, infrared spectroscopy, and electron optical techniques after heating at 50°C intervals to 700°C. Palygorskite ‘anhydride’ formed at 400°C and sillimanite formed at 500°C. The palygorskite is believed to have formed in joints in limestone of Upper Oligocene age (Duntroonian) prior to uplift and subsequent weathering. It is highly crystalline, and appears not to have altered or weathered since precipitation.

Root-knot nematodes represent a serious threat to world coffee production, especially Meloidogyne incognita and M. paranaensis. Most cultivars of Coffea arabica are highly susceptible to these parasites and cultivation in infested areas has only been possible with the use of resistant C. canephora rootstocks. In this research, three elite clones of C. canephora, selected in areas infested by M. incognita and M. paranaensis, were evaluated in controlled conditions to assess levels of resistance against two populations of M. paranaensis, four populations of M. incognita and a mixed population of both species. The three clones were resistant to both species, but CcK1 and CcR2 were considered most promising because their vegetative growth was not impaired by nematodes.

Cover crop–based, organic rotational no-till (CCORNT) corn and soybean systems have been developed in the mid-Atlantic region to build soil health, increase management flexibility, and reduce labor. In this system, a roller-crimped cover crop mulch provides within-season weed suppression in no-till corn and soybean. A cropping system experiment was conducted in Pennsylvania, Maryland, and Delaware to test the cumulative effects of a multitactic weed management approach in a 3-yr hairy vetch/triticale–corn–cereal rye–soybean–winter wheat CCORNT rotation. Treatments included delayed planting dates (early, intermediate, late) and supplemental weed control using high-residue (HR) cultivation in no-till corn and soybean phases. In the no-till corn phase, HR cultivation decreased weed biomass relative to the uncultivated control by 58%, 23%, and 62% in Delaware, Maryland, and Pennsylvania, respectively. In the no-till soybean phase, HR cultivation decreased weed biomass relative to the uncultivated treatment planted in narrow rows (19 to 38 cm) by 20%, 41%, and 78% in Delaware, Maryland, and Pennsylvania, respectively. Common ragweed was more dominant in soybean (39% of total biomass) compared with corn (10% of total biomass), whereas giant foxtail and smooth pigweed were more dominant in corn, comprising 46% and 22% of total biomass, respectively. Common ragweed became less abundant as corn and soybean planting dates were delayed, whereas giant foxtail and smooth pigweed increased as a percentage of total biomass as planting dates were delayed. At the Pennsylvania location, inconsistent termination of cover crops with the roller-crimper resulted in volunteer cover crops in other phases of the rotation. Our results indicate that HR cultivation is necessary to achieve adequate weed control in CCORNT systems. Integration of winter grain or perennial forages into CCORNT systems will also be an important management tactic for truncating weed seedbank population increases.

Oxyfluorfen (0.28, 0.42, 0.56, and 0.84 kg ai ha−1) under clear polyethylene film was evaluated for weed control, crop injury, and effects on yields in transplanted muskmelon, cucumber, and summer squash. Numerous narrowleaf and broadleaf weeds were effectively suppressed by 0.42 ha−1 of oxyfluorfen. Crop injury, occurring soon after transplanting, was reported in New York and North Carolina. Injury was usually transient, and injured crops frequently grew more vigorously than those grown on untreated black polyethylene mulch. Muskmelons were consistently the most tolerant of the three crops. At high rates, yields of squash and cucumber in 1988 were reduced in New York and North Carolina, respectively. In greenhouse studies, positioning the cotyledons under the polyethylene film caused greater injury in all three crops than when cotyledons remained above the plastic.