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.

For this study, we adapted the Montgomery Borgatta Caregiver Burden Scale, used widely in the United States, to the Saudi Arabian context. To produce an Arabic, culturally sensitive version of the scale, we conducted semi-structured interviews with 20 Saudi family caregivers. The Arabic version of the scale was tested, and participants were asked to comment on the appropriateness of items for the construct of “caregiver burden” using the repertory grid technique and laddering procedure – two constructivist methods derived from personal construct theory. From interview findings, we examined the content of the items and the caregiver burden construct itself. Our findings suggest that the use of constructivist methods to refine constructs and quantitative instruments is highly informative. This strategy is feasible even when little is known about the investigated constructs in the target culture and further elucidates our understanding of cross-cultural variations or invariance of different versions of the scale.

Objectives: The purpose of this study was to investigate the longitudinal trajectory of self- and informant-subjective cognitive complaints (SCC), and to determine if SCC predict longitudinal changes in objective measures (OM) of cognitive function. Methods: The study included healthy and cognitively normal late middle-aged adults enriched with a family history of AD who were evaluated at up to three visits over a 4-year period. At each visit (Visit 1–3), self- and informant-SCC and OM were evaluated. Linear mixed models were used to determine if the longitudinal rate of change of self- and informant-SCC were associated with demographic variables, depressive symptoms, family history (FH), and apolipoprotein epsilon 4 (APOE4) status. The same modeling approach was used to examine the effect of Visit 1 SCC on longitudinal cognitive change after controlling for the same variables. Results: At Visit 1, more self-SCC were associated with fewer years of education and more depressive symptoms. SCC were also associated with poorer performance on cognitive measures, such that more self-SCC at Visit 1 were associated with poorer performance on memory and executive functioning measures at Visit 1, while more informant-SCC were associated with faster rate of longitudinal decline on a measure of episodic learning and memory. FH and APOE4 status were not associated with SCC. Discussion: Self- and informant-SCC showed an association with OM, albeit over different time frames in our late middle-aged sample. Additional longitudinal follow-up will likely assist in further clarifying these relationships as our sample ages and more pronounced cognitive changes eventually emerge. (JINS, 2017, 23, 617–626)

When solid stands of foxtail millet are cut for swath grazing, the grazing season for cattle is extended and winter feeding costs reduced. The economic success of this practice depends on inexpensive weed control. Eight single- and double-rate herbicide combinations (g ai/ha) were evaluated and compared to a weed-free check: MCPA + bromoxynil (280 + 280 or 560 + 560); MCPA + clopyralid (560 + 100 or 1120 + 200); MCPA + fluroxypyr (562 + 108 or 1124 + 216); and MCPA + clopyralid + fluroxypyr (560 + 100 + 144 or 1120 + 200 + 288). This study was conducted at Indian Head, Saskatchewan (SK) in 2004, 2005, 2006, and 2007 and at Scott, SK, in 2006 and 2007. Crop injury and dry matter yield were measured. Results of the study indicate that crop injury exceeded 20% at only one out of six sites at 7 to 14 d after herbicide treatment. Double-rate MCPA + bromoxynil treatments had higher injury ratings than the weed-free check 7 to 14 d after treatment in all site years. As the growing season progressed, injury ratings tended to decline except at Scott in 2007, where injury ratings at the 21 to 35 d period were numerically greater than the other two rating periods. There were no differences among treatments for crop biomass production. We conclude that all four herbicide combinations at the labeled rate are safe to use on foxtail millet in Saskatchewan, and probably in other areas with similar environmental growing conditions.

This study determined whether surrogate organisms can predict activity against Clostridium difficile spores and compared the efficacy of hand hygiene preparations against C. difficile. Our data suggest that surrogate organisms were not predictive of C. difficile spore removal. Four preparations were significantly more effective than tap water at removing C. difficile.

The effects of vascular rarefaction (the loss of small arteries) on the circulation of blood are studied using a multiscale mathematical model that can predict blood flow and pressure in the systemic and pulmonary arteries. We augmented a model originally developed for the systemic arteries by Olufsen and coworkers and Ottesen et al. (2004) to (a) predict flow and pressure in the pulmonary arteries, and (b) predict pressure propagation along the small arteries in the vascular beds. The systemic and pulmonary arteries are modelled as separate bifurcating trees of compliant and tapering vessels. Each tree is divided into two parts representing the ‘large’ and ‘small’ arteries. Blood flow and pressure in the large arteries are predicted using a nonlinear cross-sectional-area-averaged model for a Newtonian fluid in an elastic tube with inflow obtained from magnetic resonance measurements. Each terminal vessel within the network of the large arteries is coupled to a vascular bed of small ‘resistance’ arteries, which are modelled as asymmetric structured trees with specified area and asymmetry ratios between the parent and daughter arteries. For the systemic circulation, each structured tree represents a specific vascular bed corresponding to major organs and limbs. For the pulmonary circulation, there are four vascular beds supplied by the interlobar arteries. This paper presents the first theoretical calculations of the propagation of the pressure and flow waves along systemic and pulmonary large and small arteries. Results for all networks are in agreement with published observations. Two studies were done with this model. First, we showed how rarefaction can be modelled by pruning the tree of arteries in the microvascular system. This was done by modulating parameters used for designing the structured trees. Results showed that rarefaction leads to increased mean and decreased pulse pressure in the large arteries. Second, we investigated the impact of decreasing vessel compliance in both large and small arteries. Results showed that the effects of decreased compliance in the large arteries far outweigh the effects observed when decreasing the compliance of the small arteries. We further showed that a decrease of compliance in the large arteries results in pressure increases consistent with observations of isolated systolic hypertension, as occurs in ageing.

Changes in pre- and post-fire biomass of hypogeous fungal sporocarps remaining at locations where an endangered mycophagous marsupial, the northern bettong (Bettongia tropica), had foraged, were assessed in fire-prone sclerophyll forest in northeastern Australia. Low to medium intensity experimental fires were set during the late dry season in 1995 and 1996 and post-foraging biomass of sporocarps (expressed as biomass of sporocarps remaining at recent B. tropica diggings) was measured at unburnt and burnt sites at approximately six-week intervals for a period of 14 months. Post-foraging biomass was significantly higher at burnt sites immediately following fire compared with control sites, solely due to increased biomass of hypogeous species belonging to the family Mesophelliaceae. Several months after fire, post-foraging biomass was significantly higher on unburnt sites compared with very low biomass on burnt sites. Twelve months after fire, the biomass on burnt and unburnt sites was not significantly different, having returned to biomass observed pre-fire. All evidence points toward mesophellioid fungi being greatly more available to bettongs on recently burnt ground, but fire may make several other sporocarp taxa considerably less available several months following fire.