Japanese hop (Humulus japonicus Siebold & Zucc.) is an emerging invasive plant that has been observed to invade and spread throughout wetlands. As an annual vine, H. japonicus can smother native vegetation, forming dense stands and reducing biodiversity. At a restored floodplain forest in Joslin, IL, formerly used as an experimental site to test the effectiveness of different reforestation methods, H. japonicus has invaded stands of the previously dominant invasive, reed canarygrass (Phalaris arundinacea L.). We conducted an observational field study to examine the spatiotemporal dynamics of H. japonicus invasion relative to gradients in canopy cover and species composition. Ten transects, with half the transect extending into and half extending beyond H. japonicus patches, were established in October 2022. Seven quadrats per transect were surveyed for vegetation cover and canopy cover in October 2022, June 2023, and October 2023. Transects were evenly split between forested and open areas based on the reforestation treatments. Humulus japonicus cover significantly increased from October 2022 to October 2023, resulting in a slight decrease and replacement of P. arundinacea across the site. Shade reduced H. japonicus cover, indicating its preference for sunlit conditions. Species richness was higher in forested transects compared with open ones, most likely due to the absence of both P. arundinacea and H. japonicus in shaded transects. Along transects, quadrats that had been invaded by H. japonicus differed in species composition from quadrats that had not been invaded in both October 2022 and October 2023. Humulus japonicus cover was much lower in June than October, suggesting that temporal niche partitioning may allow P. arundinacea to persist, and indicating that monitoring for H. japonicus should occur late in the growing season. Both invasive species are shade intolerant, suggesting that planting fast-growing trees should be an effective long-term solution for controlling invasion.

OBJECTIVES/GOALS: Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system. MS affects more than two million people worldwide, resulting in physical disability, cognitive impairment, and decreased quality of life. We are investigating the role of senescent cells, a hallmark of aging, in inflammation and disease progression in MS. METHODS/STUDY POPULATION: Female mice on a C57BL/6 background were subjected to the Hooke Laboratory EK-2110 experimental autoimmune encephalomyelitis protocol (EAE; n=10) to induce hind limb paralysis, and matching control mice received no injections (naīve; n=10). Immunofluorescent staining was used to visualize senescence cells and their localization in spinal cord tissue sections from naīve and EAE mice based on antibody detection of cell surface or intracellular proteins. Tissue sections from each group were analyzed in duplicates for each antibody (n=3-4/group). Flow cytometry was performed to immunophenotype the senescence cells using conjugated fluorescent antibodies to cell surface or intracellular proteins (n=5/group). RESULTS/ANTICIPATED RESULTS: Immunostaining demonstrated an increase in the cell senescence markers p16 (15-fold; unpaired t-test, p=0.01; n=3) and p21 (15-fold; unpaired t-test, p=0.003; n=3) in the spinal cord of EAE mice compared to naīve mice. Next, we showed that p16+ and p21+ cells increase with disease progression in the meninges adjacent to the ventral demyelinating lesions (one-way ANOVA, p=0.0009; n=3/group). We further found that 30±9.7% of M2 macrophages, a subset of myeloid cells, express p21 in the spinal cord of EAE mice by using flow cytometry analysis compared to only 5±1.6% in naīve mice (unpaired t-test, p=0.002; n=5/group). DISCUSSION/SIGNIFICANCE: Our findings demonstrate that senescent cells accumulate in the meningeal compartment following EAE induction, suggesting that decreasing senescent cell burden is a promising avenue in delaying disease progression and preventing neuroinflammation in MS.

ABSTRACT IMPACT: This study highlights the importance of creatine in developmental myelination and remyelination to investigate whether creatine provides a therapeutic value during a central nervous system (CNS) demyelinating insult with a potential value in patients with Multiple Sclerosis. OBJECTIVES/GOALS: Creatine is vital for ATP buffering in the brain. Interestingly, the cells that generate myelin express the main enzyme for creatine synthesis, Gamt. Patients with Gamt mutations display intellectual delays and impaired myelination. Therefore, we hypothesize that creatine is essential for developmental myelination and improves remyelination. METHODS/STUDY POPULATION: To investigate these hypotheses, we developed a new transgenic mouse model with LoxP sites flanking exons 2-6 of the guanidinoacetate methyltransferase (Gamt) gene where excision leads to expression of a green fluorescent tag allowing us to track the cells normally expressing Gamt. We used immunohistochemistry techniques to look at the corpus callosum, the main white matter tract in the brain, and evaluate the number of oligodendrocytes (OL), glial cells responsible for generating myelin. We also used the cuprizone model of toxic demyelination to investigate whether dietary creatine and cyclocreatine, a planar analog of creatine that more efficiently crosses the blood-brain barrier, can enhance remyelination. RESULTS/ANTICIPATED RESULTS: In this mouse model, we show a 95% (+/-0.47%, n=3) co-localization of Gamt within mature OL during postnatal (P) day P14 with no co-localization in neurons or other glial cells. This suggests that mature OL are the main cells making creatine in the CNS. Next, we show that knocking out Gamt leads to a significant reduction in OL in the developing corpus callosum, at P14 and P21 (P14: 0.007, n=3; P21: 0.04, n=3). We also show that creatine supplementation causes a trending increase in mature OL density in the corpus callosum following cuprizone demyelination (2% creatine; p=0.052; n=4). Interestingly, cyclocreatine supplementation significantly increased mature OL density in the corpus callosum following cuprizone demyelination (0.1% cyclocreatine; p=0.034; n=4). DISCUSSION/SIGNIFICANCE OF FINDINGS: These studies highlight the important role creatine plays in developmental myelination and remyelination to investigate whether creatine and cyclocreatine provide a therapeutic value during a CNS demyelinating insult. This work investigates a potential therapeutic value of creatine to patients with Multiple Sclerosis.

OBJECTIVES/GOALS: Oligodendrocytes (OL) are glial cells of the central nervous system (CNS) responsible for the energy demanding task of generating myelin sheaths during development and remyelination after demyelinating injury. One metabolite shown to significantly increase ATP production in OL is the nitrogenous organic acid, creatine. Creatine plays an essential role in ATP buffering within tissues with highly fluctuating energy demands such as brain and muscle. Interestingly, mature OL, which are the cells capable of myelin production, are the main cells in the CNS expressing the rate-limiting enzyme for creatine synthesis, guanidinoacetate methyltransferase (Gamt). Patients with mutations in Gamt display intellectual disabilities, impaired myelination and seizures. Therefore, we hypothesize that creatine may be essential for developmental myelination and improve remyelination. METHODS/STUDY POPULATION: To investigate these hypotheses, we developed a new transgenic mouse model with LoxP sites flanking exons 2-6 of the Gamt gene where excision leads to expression of a green fluorescent tag allowing us to track the cells normally expressing Gamt. RESULTS/ANTICIPATED RESULTS: In this mouse model, we show a 95% (±0.47%, n = 3) co-localization of Gamt within mature OL during postnatal (P) day P14. Next, we show that knocking out Gamt leads to a significant reduction in OL in the major CNS white matter tract, the corpus callosum, at P14 and P21 (P14: 0.007, n = 3; P21: 0.04, n = 3). Here, we also investigate whether dietary creatine can enhance remyelination in the cuprizone model of toxic demyelination. DISCUSSION/SIGNIFICANCE OF IMPACT: These studies highlight the important role creatine plays in developmental myelination and investigate whether creatine can provide a therapeutic value during a CNS demyelinating insult.

With the LAMOST DR4 and Gaia DR2 common red clump giant stars, we investigate the three-dimensional kinematics of Milky Way disk stars in mono-age populations between Galactocentric distances of R = 6 and 15 kpc. We confirm the 3D asymmetrical motions of recent works, and provide time tagging of the Galactic outer disk asymmetrical motions. Radial motions present a north-south asymmetry in the region corresponding to recent density and velocity substructures that were sensitive to the perturbations in the early 6 Gyr. What’s more, we discover a new velocity substructure in the north side corresponding to density dip found recently (“south-middle opposite”) in the radial and azimuthal velocity. Meanwhile, the vertical velocity with clear vertical bulk motions or bending mode motions has no clear asymmetry corresponding to the in-plane asymmetrical features.

OBJECTIVES/SPECIFIC AIMS: The overall objective of this proposal is to establish and modulate the inflammatory profile of individuals across the spectrum of multiple sclerosis (MS), with a focus on determining the potential of interleukin 4-induced protein 1 (IL4I1) as a possible marker of progression and modulator of inflammation in human blood samples. METHODS/STUDY POPULATION: The proposed experimental approach involves isolating plasma and peripheral blood mononuclear cells (PBMCs) from individuals across the spectrum of MS phenotypes, and analyzing these samples primarily by quantitative polymerase chain reaction (qPCR) and enzyme-linked immunosorbent assay (ELISA) methods. Specifically, study groups include: (1) actively relapsing-remitting MS (a-RRMS), (2) non-actively relapsing-remitting MS (n-RRMS), (3) non-active secondary-progressive MS (SPMS), (4) other autoimmune diseases (OAD), (5) healthy controls (HC). RESULTS/ANTICIPATED RESULTS: We expect that IL4I1 treatment increases regulatory cytokine (eg, IL10, TGFb) expression while decreasing Th1 and Th17-derived cytokines (IFNg, IL17), as well as increasing relative composition of regulatory cells (Th2, Treg, M2) as compared with Th1, Th17, M1 (aim 1). Preliminary data on healthy control cells support this prediction. Our central hypothesis is that IL4I1 level indicates the body’s ability to repair itself. As such, we anticipate that all MS groups are deficient in IL4I1, to varying degrees, such that HC>n-RRMS>a-RRMS>SPMS. HC have full repair capacity. RRMS>SPMS as remission indicates existent repair capacity, which is lost in SPMS. n-RRMS>a-RRMS since both, as RRMS, capable of repair response, but a-RRMS triggered this response more recently in response to more recent relapse. In all groups, we expect IL4I-treatment to mitigate inflammation (aim 2). Finally, we expect that H2O2 production by IL4I1 is a key player in IL4I1 function, and that H2O2 will preferentially induce oxidative stress to pro-inflammatory subsets of PBCMs (aim 3). DISCUSSION/SIGNIFICANCE OF IMPACT: MS is a chronic inflammatory neurodegenerative disease of the central nervous system that, with an average age of onset of 34, afflicts over 2.3 million individuals worldwide during many of the most productive years of their lives. The pathogenesis of MS, which involves autoimmune destruction of myelin, is poorly understood. Accurate biomarkers, which could predict disease progression, are yet to be identified and would provide valuable information to patients and their treating clinicians. Likewise, effective treatments are few and in high demand. IL4I1 is a promising candidate for both roles.

In many jurisdictions, legislation requires long-term care (LTC) facilities to have a registered nurse on duty 24 hours a day, seven days per week. Although considerable research exists on LTC nurse staffing intensity, no empirical research on this requirement exists. Our retrospective observational study in Saskatchewan compared facilities with 24/7 RN coverage to facilities with less coverage supplemented with various night-shift staffing models. Adjusted for nurse staffing intensity and other potential confounders, risk ratios associated with less-than-24/7 RN coverage supplemented with licensed practical nurse night-shift staffing were 1.17, 95% CI [0.91, 1.50] and 1.00, 95% CI [0.72, 1.39]; and with less-than-24/7 RN coverage supplemented with care aide night-shift staffing, risk ratios were 1.46, 95% CI [1.11, 1.91] and 1.11, 95% CI [0.78, 1.58], for inpatient hospital admissions and Emergency Department visits respectively. Findings suggest that acute services utilization may be negatively influenced by the absence of 24/7 RN coverage.

This paper explores the relationship between digital and material-based fabrications in architecture. The notion of transient materialization proposes that immaterial architecture is a trigger for investigating new possibilities for digital fabrication through space and time. This project is mainly inspired by the beauty of nature, focusing on soap foam bubbles, which have an n-hedron structure and are usually blown by air. The paper questions this structure’s materiality, examines its physical performance and ephemeral characteristics, and expands on its meaning through an experiment in digital fabrication. In addition, we present the first phase of this technology, in which an anti-gravity and programmable foam structure was achieved. The experiment demonstrates the different shapes possible for dynamic and transient soap foam structures.