OBJECTIVES/GOALS: In triple negative breast cancer (TNBC), obesity is associated with poor outcomes. Adipose stem cells (ASCs) from obese patients (obASCs) secrete higher levels of adipokines compared to ASCs from lean individuals. Leptin, one of these adipokines, has been implicated in many cancers. This study seeks to examine the role of leptin signaling in TNBC. METHODS/STUDY POPULATION: Previous work in conjunction with a collaborating lab has shown that leptin signaling promotes metastasis and increased expression of epithelial-mesenchymal transition (EMT) markers in triple negative breast cancer cell lines. This project expands upon this work through using both patient-derived cell lines and and patient-derived xenografts (PDX), and examines the role of leptin signaling both in vitro and in vivo. To determine the effects of obesity upon a PDX model of TNBC, a high fat diet was used to induce obesity in vivo. A pharmacological inhibitor of the leptin receptor was used to test the requirement for leptin signaling both in vivo and in vitro. RESULTS/ANTICIPATED RESULTS: Exposure to conditioned media harvested from obASCs increased the percentage of TNBC cells that expressed cancer stem cell markers, whereas exposure to an inhibitor of the leptin receptor decreased the percentage of cells with cancer stem cell markers. PDX tumors implanted into mice with diet-induced obesity had an increased volume compared to tumors implanted into lean controls. Further analysis will be conducted on metastasis, circulating tumor cells, and survival in both lean and obese mice. DISCUSSION/SIGNIFICANCE: Understanding the complex signaling events in the obese tumor microenvironment is essential, as these molecular differences may contribute to different outcomes for obese and lean individuals with triple negative breast cancer. Therefore, study of the crosstalk between obASCs and TNBC cells is critical.

OBJECTIVES/GOALS: Current approaches to drug development for the aggressive triple negative breast cancer rely on current 2D and 3D in vitro models which have limited capabilities. We have developed a translational microphysiological system that can maintain the human breast microenvironment to capture the complex interaction with the tumor microenvironment. METHODS/STUDY POPULATION: Three different TNBC cell lines were seeded in BC-MPS: MDA-MB-231 parental cell line, MDA-MB-231wiht the gene, LKB1 overexpressed, which is a tumor suppressor, and MDA-MB-231 with the enzyme, ERK5, an enzyme associated with increased metastasis and drug resistance, knocked out. These three TNBC cell lines were cultured in a standard 2D 96-well plate and in BC-MPS. Time-lapse videos were taken to track cellular mobility. RNA-sequencing was performed to compare different expression levels of various cancer related genes of the cell lines cultured in standard 2D and BC-MPS. RESULTS/ANTICIPATED RESULTS: The LKB1 overexpressed MDA-MB-231 and the ERK5-ko MDA-MB-231 cell lines are expected to have decreased mobility compared to the parental cells. The cell lines are expected to have increased expression of cancer related genes when cultured in BC-MPS than when cultured in standard 2D due to the presence of human breast tissue. DISCUSSION/SIGNIFICANCE: BC-MPS is a promising new translational MPS that facilitates studying long term interactions between real human breast tissue and cancer cells. The BC-MPS systems ability to support the growth of established cell lines has been demonstrated. Future studies will focus on developing the model for personalized medicine.

Stem cells give rise to the entirety of cells within an organ. Maintaining stem cell identity and coordinately regulating stem cell divisions is crucial for proper development. In plants, mobile proteins, such as WUSCHEL-RELATED HOMEOBOX 5 (WOX5) and SHORTROOT (SHR), regulate divisions in the root stem cell niche. However, how these proteins coordinately function to establish systemic behaviour is not well understood. We propose a non-cell autonomous role for WOX5 in the cortex endodermis initial (CEI) and identify a regulator, ANGUSTIFOLIA (AN3)/GRF-INTERACTING FACTOR 1, that coordinates CEI divisions. Here, we show with a multi-scale hybrid model integrating ordinary differential equations (ODEs) and agent-based modeling that quiescent center (QC) and CEI divisions have different dynamics. Specifically, by combining continuous models to describe regulatory networks and agent-based rules, we model systemic behaviour, which led us to predict cell-type-specific expression dynamics of SHR, SCARECROW, WOX5, AN3 and CYCLIND6;1, and experimentally validate CEI cell divisions. Conclusively, our results show an interdependency between CEI and QC divisions.

ABSTRACT IMPACT: Identifying an important pathway in treatment resistant TNBC will allow for the future development of clinical therapeutics specific for this disease. OBJECTIVES/GOALS: Triple Negative Breast Cancer (TNBC) is a subtype of breast cancer characterized by negative expression of estrogen receptor, progesterone receptor, and HER2/neu amplification. It resists therapies and has a high recurrence rate after resection. The goal of my research is to identify & characterize a TNBC pathway for future development of therapies. METHODS/STUDY POPULATION: The project uses a combination of cell lines, patient derived xenograft (PDX) models, as well as patient databases. Standard cellular and molecular biology techniques will be used including: Cell culture, qPCR, western blotting, and flow cytometry. RESULTS/ANTICIPATED RESULTS: LKB1 is a master kinase that activates 14 possible downstream kinases. The signaling pathway has been demonstrated to play a role in energy homeostasis and metabolism. Mutation of LKB1 signaling results in Peutz-Jeghers Syndrome and is associated with neoplasias of the lung, pancreas, and breast. Based on preliminary analysis, overexpression of LKB1 by shRNA in TNBC cell lines results in suppression of EMT and reduction of the cancer stem cell population. Additional studies show that LKB1 overexpression has no effect on growth rate in 2D culture while significant reduction in 3D mammosphere formations can be seen. Downstream studies using commercially available SIK1 inhibitor HG-9-91-01 is able to induce a larger fraction of CSC from reduced LKB1 overexpression as well as from baseline levels. DISCUSSION/SIGNIFICANCE OF FINDINGS: Overall, our results suggest that LKB1 acts through SIK1 to suppress EMT and the generation of cancer stem cells. This results in reduced cancer functionality, as evidenced by inhibition of mammosphere formation. These results establishes a foundation for future mechanistic studies on the LKB1 axis and its mechanisms in TNBC.

Early detection of breast cancer is required to increase the chances of a successful treatment. However, current breast-imaging systems such as X-Ray mammography, breast ultrasound, and magnetic resonance imaging have technological limitations so that novel solutions are needed to address this major societal problem. The current paper considers ultra-wideband (UWB) microwave radiation in the frequency band from 1 to 9 GHz. Given by the non-ionizing nature of microwaves frequent check-ups are more feasible. In this work, we propose algorithms for qualitative and quantitative microwave breast imaging for a transmission-based UWB system. Based on numerical and experimental data, the performance of the algorithms has been investigated and compared. Finally, microwave images obtained during an initial patient study are discussed relative to corresponding X-ray images.

We introduce the asprilo1 framework to facilitate experimental studies of approaches addressing complex dynamic applications. For this purpose, we have chosen the domain of robotic intra-logistics. This domain is not only highly relevant in the context of today's fourth industrial revolution but it moreover combines a multitude of challenging issues within a single uniform framework. This includes multi-agent planning, reasoning about action, change, resources, strategies, etc. In return, asprilo allows users to study alternative solutions as regards effectiveness and scalability. Although asprilo relies on Answer Set Programming and Python, it is readily usable by any system complying with its fact-oriented interface format. This makes it attractive for benchmarking and teaching well beyond logic programming. More precisely, asprilo consists of a versatile benchmark generator, solution checker and visualizer as well as a bunch of reference encodings featuring various ASP techniques. Importantly, the visualizer's animation capabilities are indispensable for complex scenarios like intra-logistics in order to inspect valid as well as invalid solution candidates. Also, it allows for graphically editing benchmark layouts that can be used as a basis for generating benchmark suites.

Let K be a number field or a function field of characteristic 0, let φ ∈ K(z) with deg(φ) ⩾ 2, and let α ∈ ℙ1(K). Let S be a finite set of places of K containing all the archimedean ones and the primes where φ has bad reduction. After excluding all the natural counterexamples, we define a subset A(φ, α) of ℤ⩾0 × ℤ>0 and show that for all but finitely many (m, n) ∈ A(φ, α) there is a prime 𝔭 ∉ S such that ord𝔭(φm+n(α)−φm(α)) = 1 and α has portrait (m, n) under the action of φ modulo 𝔭. This latter condition implies ord𝔭(φu+v(α)−φu(α)) ⩽ 0 for (u, v) ∈ ℤ⩾0 × ℤ>0 satisfying u < m or v < n. Our proof assumes a conjecture of Vojta for ℙ1 × ℙ1 in the number field case and is unconditional in the function field case thanks to a deep theorem of Yamanoi. This paper extends earlier work of Ingram–Silverman, Faber–Granville and the authors.

Introduction/Innovation Concept: Free Open Access Medical education (FOAM) is a rapidly emerging medium for the dissemination of medical knowledge, especially in Emergency Medicine. However, the most contributors to FOAM are EM attendings who write on established platforms which they also maintain. EM learners have difficulty breaking into this quickly evolving field. In an effort to encourage FOAM involvement of trainees early in their careers, CanadiEM recruited 10 junior residents and medical students with the purpose of developing the skills necessary to contribute to FOAM. These Junior Editors actively participate in the blog workflow, developing writing, editorial, and management skills necessary to operate a high-traffic EM website. Methods: Potential candidates were recruited by placing an advertisement and application on the CanadiEM website. 10 medical students or junior residents were invited to online group video interviews and were all accepted as Junior Editors (JE). Senior CanadiEM staff held online training sessions for all new JEs on how to use Wordpress to create, edit and publish posts, as well as basics in Search Engine Optimization. The junior editors collaboratively developed an instructional document containing the information they learned during these sessions. JEs then volunteered for editorial jobs via an online messaging system (Slack) as they became available. After uploading the draft of each post, the final products are reviewed by senior Editor and feedback was given to each JE. Curriculum, Tool, or Material: All JEs have learned to use the Wordpress blogging platform to create, edit, and upload posts; optimize blog posts for search engines. Following their own interests, some JEs have also learned to edit podcasts, promote the blog on social media resources (Twitter and Facebook), create infographics, and copy-edit blog posts. Conclusion: After 8 months, the JE program has yielded 6 very active editors who maintain a strong blog workflow, have well-developed social media skills, and are actively involved in developing their own content for future posts. The JE program is a strong pathway to introduce medical trainees to both the technical and creative aspects of FOAM and serves as a novel approach to transition students from passive utilization of online content to active contributors.

With Version 2 of the high-efficiency video coding standard, a new compression efficiency tool targeting redundancies among color components is specified for all 4:4:4 profiles, and referred to as cross-component prediction (CCP). This paper describes and analyses two additional extensions to the specified CCP variant. In the first extension, an additional predictor is introduced. Particularly, beside the luma component, also the first chroma component can serve as a reference for prediction of the second chroma component. The second extension proposes a method for predicting the CCP model parameter from the statistics of already reconstructed neighboring blocks. A performance analysis of coding RGB content in different color representations is given in comparison with CCP and both extensions. Experimental results show that the proposed extensions can improve the compression efficiency effectively compared with CCP, when applied in the YCbCr domain.

This article describes a formal proof of the Kepler conjecture on dense sphere packings in a combination of the HOL Light and Isabelle proof assistants. This paper constitutes the official published account of the now completed Flyspeck project.

Transition metal dichalcogenides such as WS2 show exciting promise in electronic and optoelectronic applications. Significant variations in the transport, Raman, and photoluminescence (PL) can be found in the literature, yet it is rarely addressed why this is. In this report, Raman and PL of monolayered WS2 produced via different methods are studied and distinct features that indicate the degree of crystallinity of the material are observed. While the intensity of the LA(M) Raman mode is found to be a useful indicator to assess the crystallinity, PL is drastically more sensitive to the quality of the material than Raman spectroscopy. We also show that even exfoliated crystals, which are usually regarded as the most pristine material, can contain large amounts of defects that would not be apparent without Raman and PL measurements. These findings can be applied to the understanding of other two-dimensional heterostructured systems.

In this paper, we report on a multifunctional battery assembly, which possesses a balanced combination of energy storage capability and resistance to electrical failure under mechanical impact loading. The Granular Battery Assembly (GBA) presented here exhibits a mechanical response that emerges from features of granular and cellular media. We demonstrate that for the specific GBA embodiment considered in the present study, the electrical reliability following a mechanical loading event is substantively increased compared to that of plain battery cells. The increased reliability is due to the sacrificial material elements interspersed between the battery units, attributing energy absorption and local stress limiting.