Sorption of K from mixed KCl and CaCl2 solutions by K-depleted, Ca-saturated phlogopite was studied to determine the effect of particle size. The experiments were done at room temperature with 25 mg of K-depleted phlogopite samples in 50 ml solutions which were 0·002 N with respect to KCl and 0·02 N with respect to CaCl2.

Sorption of K increased sharply with increase in particle size. The 54–75 μm fraction sorbed nearly all, whereas the 0·2–2 μm fraction sorbed less than half, of its depleted K. The 5–20 μm fraction sorbed an intermediate amount. This relationship is explained by the same hypothesis which accounts for the increase of K release with increase in particle size. That is, bending of unit mica layers due to peripheral expansion is greater in large and thick particles than in small and thin ones. This increased bending induces the greater K release from large particles. Similarly, bending due to peripheral collapse of hydrated layers is greater in large particles than in small ones. Thus, more energy is needed to initiate layer collapse and restrict further K uptake in the large particles which results in their greater K sorption capacity.

These results imply that in natural conditions, as in soils, the coarse vermiculite and weathered mica fractions may be more effective in sorbing K from solution than their fine counterparts.

Rates of exchange of K with Ca for fine (0·2–2 μm) and coarse (54–75 μm) and for thin and thick (37–45 μm) phlogopite particles were determined using a repeated batch technique, which gave a measure of K selectivity.

Potassium selectivity of the fine fraction was higher than that of the coarse one throughout the exchange process in which 93 per cent of the total K was exchanged from the fine fraction and 100 per cent from the coarse one. Potassium selectivity of the thin 37–45 μm particles was higher initially than that of the thick 37–45 μm particles but the difference disappeared subsequently and practically 100 per cent of the total K was exchanged from both the thin and thick particles.

The results are interpreted as tentatively confirming the hypothesis that bending and deformation of elementary layers during K exchange increase with particle thickness, which in turn increase K exchange and decrease K selectivity.

The K exchange curves for the fine and coarse phlogopite fractions suggest that in natural conditions, as in soils, where K is not continuously removed from solution, vermiculization of coarse mica particles may be not only more complete but also more rapid than vermiculization of fine mica particles.

Electron probe micro-analysis studies on individual particles (40–60 mesh) of weathered micas treated with solutions containing equivalent amounts of Rb and Sr showed partial segregation of these elements. Rb was concentrated at particle and step edges, at cracks, and, in the case of partially K-depleted biotite, at boundaries of vermiculite and mica zones (“wedge zones”). The scarcity of wedge zones in mica from which nearly all of the K had been removed reduced the overall selectivity for Rb. The restricted exchange of interlayer Mg ions from vermiculite-like zones by a mixed Rb-Sr solution was observed in earlier studies with these micas. The proposed explanation for these results was a closing down of the interlayer space at the edge of the particle due to Rb concentration in these positions. This explanation is confirmed by the present study.

The selectivity of K over Ca of Amelia biotite increased sharply upon oxidation by H2O2 at pH 6·0. This increase in K selectivity was only partially reversible upon reduction by Na2S2O4. Oxidation by H2O2 of the Ca-form of this biotite resulted in a loss of no more than 2·8 and 0·4 per cent of the total Fe and Al, respectively, and caused a small and perhaps insignificant decrease in layer charge. Although 95 per cent of the structural Fe2+ of the Ca-form of this biotite was ozidized by H2O2, only 17 per cent was reduced again by four treatments with Na2S2O4.

The evidence indicates that under the conditions of this experiment the loss of protons from structural hydroxyls was the dominant mechanism by which electroneutrality in the biotite was maintained during oxidation of structural Fe2+. Because this mechanism increases the bond strength of interlayer K, it explains the increased K selectivity of biotite upon H2O2 oxidation. The relatively small reduction by Na2S2O4 of structural Fe3+ to Fe2+, which implies an equally small reprotonation of structural hydroxyls, explains the incomplete reversibility of K selectivity by Na2S2O4 treatment.

Using hydrodynamic simulations and photoionization calculations, we demonstrate that quasar emission line spectra contain information on the driving mechanism of galaxy-scale outflows. Outflows driven by a hot shocked bubble are expected to exhibit LINER-like optical line ratios, while outflows driven by radiation pressure are expected to exhibit Seyfert-like line ratios. Driving by radiation pressure also has a distinct signature in the narrow UV lines, which is detected in an HST-COS spectrum of a nearby quasar hosting a large-scale wind.

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.

OBJECTIVES/GOALS: The goal of this proposal is to develop a technology that combines calcium imaging via confocal microscopy, and force measurement via monolayer stress microscopy to perform simultaneous quantitative measurements of agonist-induced Ca2+ and mechanical signals in HASMCs. METHODS/STUDY POPULATION: The methods by which second messenger signals and changes in mechanical forces determine specific physiological responses are complex. Recent studies point to the importance of temporal and spatial encoding in determining signal specificity. Hence, approaches that probe both chemical and mechanical signals are needed. We combine hyperspectral imaging for second messenger signal measurements, monolayer stress microscopy for mechanical force measurements, and S8 analysis software for quantifying localized signals. Imaging was performed using an excitation-scanning hyperspectral microscope. Hyperspectral images were unmixed to identify signals from fluorescent labels and microparticles. Images were analyzed to quantify localized force dynamics through monolayer stress microscopy. RESULTS/ANTICIPATED RESULTS: Results indicate that localized and transient cellular signals can be quantified and mapped within cell populations. Importantly, these results establish a method for simultaneous interrogation of cellular signals and mechanical forces that may play synergistic roles in regulating downstream cellular physiology in confluent monolayers. DISCUSSION/SIGNIFICANCE: We will measure the distribution of chemical and mechanical signals within cells, providing insight into the dynamics of cell signaling. Studies will have implication in the understanding of infections, drug delivery in which non-uniform distributions of drugs are a certainty, and in understanding coordinated responses in cellular systems.

Lairage staff at 11 abattoirs were asked to rate which producers regularly provided pigs which were ‘easy’ (EH) or ‘difficult’ (DH) to handle, on a scale of one (very DH) to five (very EH). A postal questionnaire, dealing with various aspects of post-weaning farm management, was then given to the four or five producers sending the most EH and the four or five producers sending the most DH pigs to each abattoir. Of 105 questionnaires sent, information on 26 EH and 27 DH systems was returned. The median number of replies per abattoir was two for both EH and DH systems. In most systems (77%) pigs experienced three or four housing stages from weaning to slaughter. In each of the first five housing stages, more EH pigs had access to daylight (mean of 86% ± 11.5 (SD)) than DH pigs (mean of 64% ± 10.1 (SD), P < 0.05, two-sample t test). More EH systems provided straw in the first three housing stages, although over all stages the difference was not significant. During housing stage two, the difference in provision of straw between the systems was most marked, with 58 per cent of EH and 27 per cent of DH systems providing straw. Distance walked between housing stages three to four and four to five was significantly greater for EH compared to DH systems (EH mean of 64m ± 24.1 (SD), versus DH mean of 22m ± 14.0 (SD), and EH mean of 73m ±17.2 (SD), versus DH mean of 23m ± 8.5 (SD), P <0.001 and 0.01 respectively, two-sample t test). At loading for pre-slaughter transport, moving from daylight to daylight conditions occurred in 65 per cent of EH and 25 per cent of DH systems. Overall, the results provide circumstantial evidence that environmental factors can affect ease of handling, and hence pig welfare during pre-slaughter transport and lairage.

The solution to weak bureaucratic capacity in developing countries is often presumed to be more accountability. This paper shows how accountability initiatives, intended to reduce corruption, can actually hinder the development of capable government agencies by making it harder for directors to recruit experts and spend their budgets. It further highlights a common way public servants escape the accountability rules that limit their effectiveness: outsourcing bureaucracies to nonstate organizations. This practice of outsourcing bureaucracy to avoid accountability rules creates what I call “shadow” state capacity and, paradoxically, it may help explain “pockets of effectiveness” among government social programs in developing countries. Drawing on in-depth interviews and descriptive statistics, I show how outsourcing was a critical factor in producing two of Brazil’s most vaunted social sector programs. However, I also suggest that outsourcing bureaucracy may ultimately limit state capacity, even if it helps to build capable programs in the short run.

Bileaflet mechanical heart valves (BMHV) create unphysiological turbulent flow. Such turbulent flow involves multiple instability mechanisms interacting with one another in a confined geometry. For instance, an impinging leading-edge vortex (ILEV) instability creates disturbances at the leading edge of the valve leaflets, while potentially promoting turbulence downstream of the BMHV (Zolfaghari and Obrist, Phys. Rev. Fluids, vol. 4, 2019). In this article, we use adjoint-based methods to study the structural sensitivity of the ILEV instability in the BMHV, and to quantify the role of this instability in the maximum disturbance energy growth in the wake of the BMHV. We first present a direct numerical simulation to show the effect of the ILEV instability on the turbulent flow in the wake of the valve. Second, we perform a modal linear stability analysis on a two-dimensional subdomain attached to the leading edge of one leaflet. We investigate the sensitivity of the global modes associated with this flow using their adjoints, and then show a passive control scenario using a local feedback source. This results in a partial improvement in the flow oscillations downstream of the leaflets. We finally present a non-modal approach to identify the optimal initial conditions for achieving maximum energy growth at arbitrary locations. We show that, for sufficiently large times, the optimal initial condition for highest energy growth in the wake points at the leading edge, which includes the ILEV instability. Our study illustrates that an improved leading-edge shape can effectively reduce turbulence in the wake of the BMHV.

New occurrences of flask-shaped and envelope-bearing microfossils, including the predominantly Cambrian taxon Granomarginata, are reported from new localities, as well as from earlier in time (Ediacaran) than previously known. The stratigraphic range of Granomarginata extends into the Cambrian System, where it had a cosmopolitan distribution. This newly reported Ediacaran record includes areas from Norway (Baltica), Newfoundland (Avalonia) and Namibia (adjacent to the Kalahari Craton), and puts the oldest global occurrence of Granomarginata in the Indreelva Member (< 563 Ma) of the Stáhpogieddi Formation on the Digermulen Peninsula, Arctic Norway. Although Granomarginata is rare within the assemblage, these new occurrences together with previously reported occurrences from India and Poland, suggest a potentially widespread palaeogeographic distribution of Granomarginata through the middle–late Ediacaran interval. A new flask-shaped microfossil Lagoenaforma collaris gen. et sp. nov. is also reported in horizons containing Granomarginata from the Stáhpogieddi Formation in Norway and the Dabis Formation in Namibia, and flask-shaped fossils are also found in the Gibbett Hill Formation in Newfoundland. The Granomarginata–Lagoenaforma association, in addition to a low-diversity organic-walled microfossil assemblage, occurs in the strata postdating the Shuram carbon isotope excursion, and may eventually be of use in terminal Ediacaran biostratigraphy. These older occurrences of Granomarginata add to a growing record of body fossil taxa spanning the Ediacaran–Cambrian boundary.

The U.S. labor market continues to grapple with a “skills gap” (Marshall & Craig, 2019): a disconnect between the skills employers need and the number of job-seekers with those skills. Compounded by historically low unemployment rates, this gap is leaving employers with unfilled jobs and narrow talent pipelines. Concurrently, there are lingering concerns regarding underrepresentation of women and minorities in certain sectors of the labor market—particularly occupations in science, technology, engineering, and mathematics (STEM). This article examines how the traditional interest-only career guidance tools used in education significantly influence the gender-based skills gaps that persist in high-demand careers and introduces YouScience, a company that is helping ameliorate the skills gap by combining measures of aptitudes and interests in a new career discovery platform. We close by presenting action steps for students, parents, educators, and counselors, as well as positing possible effects of COVID-19 on career exploration and counseling.

Our studies of mammary tumor virus have included the application of the unlabeled antibody enzyme method of Sternberger to mammary tumor derived mouse cells in culture and observation with an electron microscope. The method avoids the extravagance of covalent binding of indicator molecules (horseradish peroxidase) with precious antibody locator molecules by relying instead upon specific antibody-antigen linkages. Our reagents included: Primary Antibody, rabbit anti-murine mammary tumor virus (MuMTV) which was antiserum 113 AV-2; Secondary Antibody, goat anti-rabbit IgG gamma chain (Cappel Laboratories); andthe Indicator, rabbit anti-horseradish peroxidase - horseradish peroxidase complex (PAP) (Cappel Labs.). Dilutions and washes were made in 0.05 M Tris 0.15 M saline buffered to pH 7.4. Cell monolayers, after light fixation in glutaraldehyde, were incubated in place by a protocol adapted from Sternberger and Graham and Karnovsky, then embedded by our usual method for monolayers. Reagents were confined to specific areas by neoprene 0-rings (Parker Seal Co.) reducing the amount of reagent needed to 50 microliters, 1/6th of that required to wet a 35 mm petri dish.