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.

The Astrobiology Graduate Conference (AbGradCon) is an annual conference both organized for and by early-career researchers, postdoctoral fellows, and students as a way to train the next generation of astrobiologists and develop a robust network of cohorts moving forward. AbGradCon 2021 was held virtually on 14–17 September 2021, hosted by the Earth-Life Science Institute (ELSI) of Tokyo Institute of Technology after postponement of the in-person event in 2020 due to the COVID-19 pandemic. The meeting consisted of presentations by 120 participants from a variety of fields, two keynote speakers, and other career-building events and workshops. Here, we report on the organizational and executional aspects of AbGradCon 2021, including the meeting participant demographics, various digital aspects introduced specifically for a virtual edition of the meeting, and the abstract submission and evaluation process. The abstract evaluation process of AbGradCon 2021 is unique in that all evaluations are done by the peers of the applicants, and as astrobiology is inherently a broad discipline, the abstract evaluation process revealed a number of trends related to multidisciplinarity of the astrobiology field. We believe that meetings like AbGradCon can provide a unique opportunity for students and early career researchers in astrobiology to experience community building, inter- and multidisciplinary collaboration, and career training and would be a welcome sight in other fields as well. We hope that this report provides inspiration and a basic roadmap for organizing future conferences in any field with similar goals.

Background: Children with pathogenic variations in SCN8A can present with early infantile epileptic encephalopathy-13, benign familial infantile seizures-5 or intellectual disability alone without epilepsy. In this case series, we discuss six children with variants in SCN8A managed at BC Children’s Hospital. Methods: We describe clinical and genetic results on six individuals with SCN8A variants identified via clinical or research next-generation sequencing. Functional consequences of two SCN8A variants were assessed using electrophysiological analyses in transfected cells. Results: Clinical findings ranged from normal development with well-controlled epilepsy to significant developmental delay with treatment-resistant epilepsy. Phenotypes and genotypes in our cohort are described in the table below. Functional analysis supported gain-of-function in P2 and loss-of-function in P4. Conclusions: Our cohort expands the clinical and genotypic spectrum of SCN8A-related disorders. We establish functional evidence for two missense variants in SCN8A, including LoF variant in a patient with intellectual disability, and autism spectrum disorder without seizures.

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During the last fifteen years there has been a paradigm shift in the continuum modelling of granular materials; most notably with the development of rheological models, such as the $\mu (I)$-rheology (where $\mu$ is the friction and I is the inertial number), but also with significant advances in theories for particle segregation. This paper details theoretical and numerical frameworks (based on OpenFOAM) which unify these currently disconnected endeavours. Coupling the segregation with the flow, and vice versa, is not only vital for a complete theory of granular materials, but is also beneficial for developing numerical methods to handle evolving free surfaces. This general approach is based on the partially regularized incompressible $\mu (I)$-rheology, which is coupled to the gravity-driven segregation theory of Gray & Ancey (J. Fluid Mech., vol. 678, 2011, pp. 353–588). These advection–diffusion–segregation equations describe the evolving concentrations of the constituents, which then couple back to the variable viscosity in the incompressible Navier–Stokes equations. A novel feature of this approach is that any number of differently sized phases may be included, which may have disparate frictional properties. Further inclusion of an excess air phase, which segregates away from the granular material, then allows the complex evolution of the free surface to be captured simultaneously. Three primary coupling mechanisms are identified: (i) advection of the particle concentrations by the bulk velocity, (ii) feedback of the particle-size and/or frictional properties on the bulk flow field and (iii) influence of the shear rate, pressure, gravity, particle size and particle-size ratio on the locally evolving segregation and diffusion rates. The numerical method is extensively tested in one-way coupled computations, before the fully coupled model is compared with the discrete element method simulations of Tripathi & Khakhar (Phys. Fluids, vol. 23, 2011, 113302) and used to compute the petal-like segregation pattern that spontaneously develops in a square rotating drum.

Dense granular flows can spontaneously self-channelise by forming a pair of parallel-sided static levees on either side of a central flowing channel. This process prevents lateral spreading and maintains the flow thickness, and hence mobility, enabling the grains to run out considerably further than a spreading flow on shallow slopes. Since levees commonly form in hazardous geophysical mass flows, such as snow avalanches, debris flows, lahars and pyroclastic flows, this has important implications for risk management in mountainous and volcanic regions. In this paper an avalanche model that incorporates frictional hysteresis, as well as depth-averaged viscous terms derived from the $\unicode[STIX]{x1D707}(I)$-rheology, is used to quantitatively model self-channelisation and levee formation. The viscous terms are crucial for determining a smoothly varying steady-state velocity profile across the flowing channel, which has the important property that it does not exert any shear stresses at the levee–channel interfaces. For a fixed mass flux, the resulting boundary value problem for the velocity profile also uniquely determines the width and height of the channel, and the predictions are in very good agreement with existing experimental data for both spherical and angular particles. It is also shown that in the absence of viscous (second-order gradient) terms, the problem degenerates, to produce plug flow in the channel with two frictionless contact discontinuities at the levee–channel margins. Such solutions are not observed in experiments. Moreover, the steady-state inviscid problem lacks a thickness or width selection mechanism and consequently there is no unique solution. The viscous theory is therefore a significant step forward. Fully time-dependent numerical simulations to the viscous model are able to quantitatively capture the process in which the flow self-channelises and show how the levees are initially emplaced behind the flow head. Both experiments and numerical simulations show that the height and width of the channel are not necessarily fixed by these initial values, but respond to changes in the supplied mass flux, allowing narrowing and widening of the channel long after the initial front has passed by. In addition, below a critical mass flux the steady-state solutions become unstable and time-dependent numerical simulations are able to capture the transition to periodic erosion–deposition waves observed in experiments.

Shallow granular avalanches on slopes close to repose exhibit hysteretic behaviour. For instance, when a steady-uniform granular flow is brought to rest it leaves a deposit of thickness $h_{stop}(\unicode[STIX]{x1D701})$ on a rough slope inclined at an angle $\unicode[STIX]{x1D701}$ to the horizontal. However, this layer will not spontaneously start to flow again until it is inclined to a higher angle $\unicode[STIX]{x1D701}_{start}$, or the thickness is increased to $h_{start}(\unicode[STIX]{x1D701})>h_{stop}(\unicode[STIX]{x1D701})$. This simple phenomenology leads to a rich variety of flows with co-existing regions of solid-like and fluid-like granular behaviour that evolve in space and time. In particular, frictional hysteresis is directly responsible for the spontaneous formation of self-channelized flows with static levees, retrogressive failures as well as erosion–deposition waves that travel through the material. This paper is motivated by the experimental observation that a travelling-wave develops, when a steady uniform flow of carborundum particles on a bed of larger glass beads, runs out to leave a deposit that is approximately equal to $h_{stop}$. Numerical simulations using the friction law originally proposed by Edwards et al. (J. Fluid Mech., vol. 823, 2017, pp. 278–315) and modified here, demonstrate that there are in fact two travelling waves. One that marks the trailing edge of the steady-uniform flow and another that rapidly deposits the particles, directly connecting the point of minimum dynamic friction (at thickness $h_{\ast }$) with the deposited layer. The first wave moves slightly faster than the second wave, and so there is a slowly expanding region between them in which the flow thins and the particles slow down. An exact inviscid solution for the second travelling wave is derived and it is shown that for a steady-uniform flow of thickness $h_{\ast }$ it produces a deposit close to $h_{stop}$ for all inclination angles. Numerical simulations show that the two-wave structure deposits layers that are approximately equal to $h_{stop}$ for all initial thicknesses. This insensitivity to the initial conditions implies that $h_{stop}$ is a universal quantity, at least for carborundum particles on a bed of larger glass beads. Numerical simulations are therefore able to capture the complete experimental staircase procedure, which is commonly used to determine the $h_{stop}$ and $h_{start}$ curves by progressively increasing the inclination of the chute. In general, however, the deposit thickness may depend on the depth of the flowing layer that generated it, so the most robust way to determine $h_{stop}$ is to measure the deposit thickness from a flow that was moving at the minimum steady-uniform velocity. Finally, some of the pathologies in earlier non-monotonic friction laws are discussed and it is explicitly shown that with these models either steadily travelling deposition waves do not form or they do not leave the correct deposit depth $h_{stop}$.

In 2008, for the first time in the history of this country, a black woman became First Lady of the United States. During Barack Obama's presidency, Michelle Obama was ever present in the public eye for her advocacy on issues related to health, military families, education, and for promoting the interests of women and girls. This article contributes to ongoing scholarly discourse, as well as extensive media coverage and analysis, regarding Obama's role as wife and first lady by critically examining how the particular model of motherhood she embraced and exhibited, a model firmly rooted in the black American community, was designed to challenge negative stereotypes of black women, maternity, and families. We address the following questions in this work: How did Obama's identity as a black woman influence the policies she championed as first lady? Does Obama's mothering relate to stereotypes of black mothers and help (re)define black motherhood, and if so, how? What does it mean to be a black mater gentis or mother of the nation? Drawing on her speeches and policy initiatives, we reveal how Michelle Obama defied dominant and oppressive stereotypes of black women and mothers while simultaneously (re)defining black womanhood and motherhood for the nation.

Background: Cervical sponylotic myelopathy (CSM) may present with neck and arm pain. This study investiagtes the change in neck/arm pain post-operatively in CSM. Methods: This ambispective study llocated 402 patients through the Canadian Spine Outcomes and Research Network. Outcome measures were the visual analogue scales for neck and arm pain (VAS-NP and VAS-AP) and the neck disability index (NDI). The thresholds for minimum clinically important differences (MCIDs) for VAS-NP and VAS-AP were determined to be 2.6 and 4.1. Results: VAS-NP improved from mean of 5.6±2.9 to 3.8±2.7 at 12 months (P<0.001). VAS-AP improved from 5.8±2.9 to 3.5±3.0 at 12 months (P<0.001). The MCIDs for VAS-NP and VAS-AP were also reached at 12 months. Based on the NDI, patients were grouped into those with mild pain/no pain (33%) versus moderate/severe pain (67%). At 3 months, a significantly high proportion of patients with moderate/severe pain (45.8%) demonstrated an improvement into mild/no pain, whereas 27.2% with mild/no pain demonstrated worsening into moderate/severe pain (P <0.001). At 12 months, 17.4% with mild/no pain experienced worsening of their NDI (P<0.001). Conclusions: This study suggests that neck and arm pain responds to surgical decompression in patients with CSM and reaches the MCIDs for VAS-AP and VAS-NP at 12 months.