OBJECTIVES/GOALS: The goal of this project is to characterize the efficacy of FOXA2 as a potential biomarker for patients with metastatic castrate-resistant prostate adenocarcinoma (CRPC) before transitioning to neuroendocrine prostate cancer (NEPC). NEPC currently has no therapeutic options and poor mechanistic understand of its origins. METHODS/STUDY POPULATION: In our study, we have utilized a multi-omics approach to characterize the potential efficacy of FOXA2 as a prognostic biomarker in numerous patient-derived castrate-resistant prostate cancer (CRPC) models. We have performed ATAC-, ChIP-, RNA-seq and proteomics to fully characterize where FOXA2 is binding genome-wide, how FOXA2 alters chromatin accessibility dynamics, identify regulatory gene targets of FOXA2, and identify FOXA2 protein-protein interactors. We have supported these findings using publicly available data from independent CRPC and NEPC patient cohorts and prostate cancer cell models. RESULTS/ANTICIPATED RESULTS: Our findings show that FOXA2 overexpression suppressed androgen signaling and promoted progression to a NEPC phenotype under short- and long-term androgen deprivation conditions, respectively. Further, FOXA2 redirected the chromatin accessibility landscape to be consistent with an NEPC gene expression program, including increased chromatin accessibility for key NEPC transcription factors. FOXA2 ChIP-seq showed FOXA2 to be bound at known NEPC driver genes and epigenetic modifiers across multiple stages of prostate cancer progression. Lastly, we discovered that FOXA2 physically interacts with key NEPC TFs and epigenetic regulators, suggesting that these FOXA2 physical interactions are required for NEPC progression. DISCUSSION/SIGNIFICANCE: This project will determine the efficacy of FOXA2 as a biomarker in advanced prostatecancer samples, which will translate as a potentially useful tool for clinicians to use for treatment of advanced prostate cancer patients.

We report on the generation and delivery of 10.2 PW peak power laser pulses, using the High Power Laser System at the Extreme Laser Infrastructure – Nuclear Physics facility. In this work we demonstrate for the first time, to the best of our knowledge, the compression and propagation of full energy, full aperture, laser pulses that reach a power level of more than 10 PW.

Trematodes are the main macroparasites in coastal waters. The most abundant and widespread form of these parasites is metacercaria. Their impact on their host fitness is considered relatively low but metacercarial larvae of some species can have deleterious effects on individuals and/or populations. This review focused on the cockle Cerastoderma edule and four species of the genus Himasthla; a common host–parasite system in marine coastal environments. Our aims were (1) to review literature concerning Himasthla continua, Himasthla elongata, Himasthla interrupta and Himasthla quissetensis in cockles; (2) to provide molecular signatures of these parasites and (3) to analyse infection patterns using a 20-year monthly database of cockle monitoring from Banc d'Arguin (France). Due to identification uncertainties, the analysis of the database was restricted to H. interrupta and H. quissetensis, and it was revealed that these parasites infect cockles of the same size range. The intensity of parasites increased with cockle size/age. During the colder months, the mean parasite intensity of a cockle cohort decreased, while infection occurred in the warmest season. No inter-specific competition between trematode parasites was detected. Furthermore, even if the intensity of H. interrupta or H. quissetensis infection fluctuated in different years, this did not modify the trematode community structure in the cockles. The intensity of infection of both species was also positively correlated with trematode species richness and metacercarial abundance. This study highlighted the possible detrimental role of Himasthla spp. in cockle population dynamics. It also revealed the risks of misidentification, which should be resolved by further molecular approaches.

Quantitative plant biology is an interdisciplinary field that builds on a long history of biomathematics and biophysics. Today, thanks to high spatiotemporal resolution tools and computational modelling, it sets a new standard in plant science. Acquired data, whether molecular, geometric or mechanical, are quantified, statistically assessed and integrated at multiple scales and across fields. They feed testable predictions that, in turn, guide further experimental tests. Quantitative features such as variability, noise, robustness, delays or feedback loops are included to account for the inner dynamics of plants and their interactions with the environment. Here, we present the main features of this ongoing revolution, through new questions around signalling networks, tissue topology, shape plasticity, biomechanics, bioenergetics, ecology and engineering. In the end, quantitative plant biology allows us to question and better understand our interactions with plants. In turn, this field opens the door to transdisciplinary projects with the society, notably through citizen science.

ABSTRACT IMPACT: Plasma and CSF are not reliable estimates of drug exposure in tissue compartments relevant for treatment and prevention of infectious diseases. OBJECTIVES/GOALS: Globally, high dose fluconazole is widely used in the management of cryptococcal meningitis. While it is known to readily penetrate into cerebrospinal (CSF), less is known about drug concentrations in brain parenchymal tissues. Similarly, distribution of fluconazole into gynecological tissues has not been robustly characterized. METHODS/STUDY POPULATION: With informed consent from next-of-kin, we conducted autopsies within 24h of death for hospitalized Ugandans receiving fluconazole for treatment or secondary prophylaxis of cryptococcal meningitis. Dosing history was abstracted from medical chart and caregiver interviews. Fluconazole concentrations were determined using high-performance liquid chromatography- tandem mass spectrometry (LC-MS/MS) in plasma, CSF, 10 brain compartments (frontal, parietal, and occipital lobes, corpus callosum, globus pallidus, hippocampus, midbrain, medulla oblongata, spinal cord, and choroid plexus) and 4 female genital compartments (cervix, vagina, ovary, and uterus), depending on tissue availability. Descriptive statistics of tissue to plasma ratios were used to describe concentrations relative to plasma. RESULTS/ANTICIPATED RESULTS: Fluconazole concentrations were measured in available tissues of 21 individuals with detectable fluconazole in plasma. Daily doses of fluconazole were 200 mg (n=4), 400 mg (n=1), 800 mg (n=4), 1200 mg (n=9) or unknown (n=3). CSF concentrations (n=10) ranged from 93-1380% (median 100%) of plasma while brain concentrations (n=3) across all 10 compartments ranged from 45% to 89% (median 69%) of plasma. In the female genital tract, cervical concentrations (n=10) were 9-78% (median 65%) of plasma and in the 2 individuals with available tissue, concentrations in vaginal, ovarian, and uterine tissues were similar to cervix, ranging from 63-105% of plasma. DISCUSSION/SIGNIFICANCE OF FINDINGS: Measuring drug concentrations directly in tissues, the presumed site of action, improves estimates of drug efficacy. While fluconazole concentrations in CSF were similar to plasma, actual brain tissues were consistently lower. Concentrations were similar between upper and lower female genital tracts, but were consistently lower than plasma.

We report on a two-arm hybrid high-power laser system (HPLS) able to deliver 2 × 10 PW femtosecond pulses, developed at the Bucharest-Magurele Extreme Light Infrastructure Nuclear Physics (ELI-NP) Facility. A hybrid front-end (FE) based on a Ti:sapphire chirped pulse amplifier and a picosecond optical parametric chirped pulse amplifier based on beta barium borate (BBO) crystals, with a cross-polarized wave (XPW) filter in between, has been developed. It delivers 10 mJ laser pulses, at 10 Hz repetition rate, with more than 70 nm spectral bandwidth and high-intensity contrast, in the range of 1013:1. The high-energy Ti:sapphire amplifier stages of both arms were seeded from this common FE. The final high-energy amplifier, equipped with a 200 mm diameter Ti:sapphire crystal, has been pumped by six 100 J nanosecond frequency doubled Nd:glass lasers, at 1 pulse/min repetition rate. More than 300 J output pulse energy has been obtained by pumping with only 80% of the whole 600 J available pump energy. The compressor has a transmission efficiency of 74% and an output pulse duration of 22.7 fs was measured, thus demonstrating that the dual-arm HPLS has the capacity to generate 10 PW peak power femtosecond pulses. The reported results represent the cornerstone of the ELI-NP 2 × 10 PW femtosecond laser facility, devoted to fundamental and applied nuclear physics research.

Overdose deaths involving prescription opioids killed more than 17,000 Americans in 2017, marking a five-fold increase since 1999. High prescribing rates of opioid analgesics have been a substantial contributor to prescription opioid misuse, dependence, overdose and heroin use. There was recognition approximately ten years ago that opioid prescribing patterns were contributing to this startling increase in negative opioid-related outcomes, and federal actions, including Medicare reimbursement reform and regulatory actions, were initiated to restrict opioid prescribing. The current manuscript is a description of those actions, the effect of those actions on opioid prescribing and related patient outcomes. We also describe our proposal of methods of expanding these efforts as an important piece to further reduce opioid-related misuse, dependence, and overdose death.

Caliochory, or seed dispersal by birds as nest material, has been reported for several species, but its effectiveness remains unclear in most cases. Darwin’s finches are traditionally regarded as seed predators, but the observation of two nests challenges this assumption by demonstrating that they can act as seed dispersers via caliochory. Darwin’s finches incorporate cotton-like materials into their nests, including seeds of Darwin’s cotton (Gossypium darwinii), a shrub endemic to the Galápagos (Ecuador). Bird nests typically break down after intense rainfall, so the seeds incorporated into nests might benefit from suitable conditions for germination. By simulating the germination conditions experienced over a 72-h period by cotton seeds in a naturally fallen nest, this study qualitatively confirms the long-term viability of at least a small fraction of the seeds at the surface of the nest. Darwin’s finches might therefore provide seed-dispersal services to Darwin’s cotton and possibly, other native and exotic plants of the Galápagos commonly incorporated into nests. However, larger confirmatory studies are needed.

Using the finite-element code Elmer, we show that the full Stokes modeling of the ice-sheet/ice-shelf transition we propose can give consistent predictions of grounding-line migration. Like other marine ice-sheet models our approach is highly sensitive to the chosen mesh resolution. However, with a grid size down to <5 km in the vicinity of the grounding line, predictions start to be robust because: (1) whatever the grid size (<5 km) the steady-state grounding-line position is sensibly the same (6 km standard deviation), and (2) with a grid-size refinement in the vicinity of the grounding line (200 m), the steady-state solution is independent of the applied perturbation in fluidity, provided this perturbation remains monotonic.

Predictions of marine ice-sheet behaviour require models able to simulate grounding-line migration. We present results of an intercomparison experiment for plan-view marine ice-sheet models. Verification is effected by comparison with approximate analytical solutions for flux across the grounding line using simplified geometrical configurations (no lateral variations, no buttressing effects from lateral drag). Perturbation experiments specifying spatial variation in basal sliding parameters permitted the evolution of curved grounding lines, generating buttressing effects. The experiments showed regions of compression and extensional flow across the grounding line, thereby invalidating the boundary layer theory. Steady-state grounding-line positions were found to be dependent on the level of physical model approximation. Resolving grounding lines requires inclusion of membrane stresses, a sufficiently small grid size (<500 m), or subgrid interpolation of the grounding line. The latter still requires nominal grid sizes of <5 km. For larger grid spacings, appropriate parameterizations for ice flux may be imposed at the grounding line, but the short-time transient behaviour is then incorrect and different from models that do not incorporate grounding-line parameterizations. The numerical error associated with predicting grounding-line motion can be reduced significantly below the errors associated with parameter ignorance and uncertainties in future scenarios.

Patients with pulmonary regurgitation after tetralogy of Fallot repair have impaired aerobic capacity; one of the reasons is the decreasing global ventricular performance at exercise, reflected by decreasing peak oxygen pulse. The aims of our study were to evaluate the impact of pulmonary valve replacement on peak oxygen pulse in a population with pure pulmonary regurgitation and with different degrees of right ventricular dilatation and to determine the predictors of peak oxygen pulse after pulmonary valve replacement.

The mean and median age at pulmonary valve replacement was 27 years. Mean pre-procedural right ventricular end-diastolic volume was 182 ml/m2. Out of 24 patients, 15 had abnormal peak oxygen pulse before pulmonary valve replacement. We did not observe a significant increase in peak oxygen pulse after pulmonary valve replacement (p=0.76). Among cardiopulmonary test/MRI/historical pre-procedural parameters, peak oxygen pulse appeared to be the best predictor of peak oxygen pulse after pulmonary valve replacement (positive and negative predictive values, respectively, 0.94 and 1). After pulmonary valve replacement, peak oxygen pulse was well correlated with left ventricular stroke and end-diastolic volumes (r=0.67 and 0.68, respectively).

Our study confirms the absence of an effect of pulmonary valve replacement on peak oxygen pulse whatever the initial right ventricular volume, reflecting possible irreversible right and/or left ventricle lesions. Pre-procedural peak oxygen pulse seemed to well predict post-procedural peak oxygen pulse. These results encourage discussions on pulmonary valve replacement in patients showing any decrease in peak oxygen pulse during their follow-up.

Whereas broad-scale Amazonian forest types have been shown to influence the structure of the communities of medium- to large-bodied vertebrates, their natural heterogeneity at smaller scale or within the terra firme forests remains poorly described and understood. Diversity indices of such communities and the relative abundance of the 21 most commonly observed species were compared from standardized line-transect data across 25 study sites distributed in undisturbed forests in French Guiana. We first assessed the relevance of a forest typology based on geomorphological landscapes to explain the observed heterogeneity. As previously found for tree beta-diversity patterns, this new typology proved to be a non-negligible factor underlying the beta diversity of the communities of medium- to large bodied vertebrates in French Guianan terra firme forests. Although the species studied are almost ubiquitous across the region, they exhibited habitat preferences through significant variation in abundance and in their association index with the different landscape types. As terra firme forests represent more than 90% of the Amazon basin, characterizing their heterogeneity – including faunal communities – is a major challenge in neotropical forest ecology.

A scanning force microscope for in situ nanofocused X-ray studies (SFINX) has been developed which can be installed on diffractometers at synchrotron beamlines allowing for the combination with various techniques such as coherent X-ray diffraction and fluorescence. The capabilities of this device are demonstrated on Cu nanowires and on Au islands grown on sapphire (0001). The sample topography, crystallinity, and elemental distribution of the same area are investigated by recording simultaneously an AFM image, a scanning X-ray diffraction map, and a fluorescence map. Additionally, the mechanical response of Au islands is studied by in situ indentation tests employing the AFM-tip and recording 2D X-ray diffraction patterns during mechanical loading.

We study the mixing properties of permutations obtained as a product of two uniformly random permutations of fixed cycle types. For instance, we give an exact formula for the probability that elements 1,2,. . .,k are in distinct cycles of the random permutation of {1,2,. . .,n} obtained as a product of two uniformly random n-cycles.

Agriculture in southern Greenland has a two-phase history: with the Norse, who first settled and farmed the region between 985ad and circa 1450ad, and with the recent reintroduction of sheep farming (1920ad to the present). The agricultural sector in Greenland is expected to grow over the next century as anticipated climate warming extends the length of the growing season and increases productivity. This article presents a synthesis of results from a well-dated 1500-year lake sediment record from Lake Igaliku, south Greenland (61°00′N, 45°26′W, 15m asl) that demonstrates the relative impacts of modern and Norse agricultural activities. Pollen, non-pollen palynomorphs (NPPs), sediment mass accumulation rates, diatoms and stable isotopes of nitrogen provide a comprehensive history of both phases of agriculture and their associated impacts on the landscape and adjacent lake. The initial colonisation of southern Greenland is marked by a loss of tree birch pollen, a rise in weed taxa, and an increase in coprophilous fungi and sediment accumulation rate consistent with land-use changes. The biological and chemical proxies within the lake, however, show only slight changes in diatom taxa, and a rise in δ15N. After the Norse demise and during the Little Ice Age, most of the markers return to pre-settlement conditions. However, the continuation of non-indigenous plant taxa suggests that the landscape did not completely return to a pre-disturbance state. After 1988, the character of the lake changed markedly: mesotrophic diatoms and N isotopes all reveal major shifts consistent with a trophic shift, together with a sharp rise in sediment accumulation rate. The post-1988 lake environment, affected by modern farming development, is unprecedented within the context of the last 1500 years. These results demonstrate the potential of lake sediment studies paired with archaeological investigations to reveal the relationship between climate, environment and human societies.