The COVID-19 pandemic accelerated the development of decentralized clinical trials (DCT). DCT’s are an important and pragmatic method for assessing health outcomes yet comprise only a minority of clinical trials, and few published methodologies exist. In this report, we detail the operational components of COVID-OUT, a decentralized, multicenter, quadruple-blinded, randomized trial that rapidly delivered study drugs nation-wide. The trial examined three medications (metformin, ivermectin, and fluvoxamine) as outpatient treatment of SARS-CoV-2 for their effectiveness in preventing severe or long COVID-19. Decentralized strategies included HIPAA-compliant electronic screening and consenting, prepacking investigational product to accelerate delivery after randomization, and remotely confirming participant-reported outcomes. Of the 1417 individuals with the intention-to-treat sample, the remote nature of the study caused an additional 94 participants to not take any doses of study drug. Therefore, 1323 participants were in the modified intention-to-treat sample, which was the a priori primary study sample. Only 1.4% of participants were lost to follow-up. Decentralized strategies facilitated the successful completion of the COVID-OUT trial without any in-person contact by expediting intervention delivery, expanding trial access geographically, limiting contagion exposure, and making it easy for participants to complete follow-up visits. Remotely completed consent and follow-up facilitated enrollment.

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.

An emergent volume electron microscopy technique called cryogenic serial plasma focused ion beam milling scanning electron microscopy (pFIB/SEM) can decipher complex biological structures by building a three-dimensional picture of biological samples at mesoscale resolution. This is achieved by collecting consecutive SEM images after successive rounds of FIB milling that expose a new surface after each milling step. Due to instrumental limitations, some image processing is necessary before 3D visualization and analysis of the data is possible. SEM images are affected by noise, drift, and charging effects, that can make precise 3D reconstruction of biological features difficult. This article presents Okapi-EM, an open-source napari plugin developed to process and analyze cryogenic serial pFIB/SEM images. Okapi-EM enables automated image registration of slices, evaluation of image quality metrics specific to pFIB-SEM imaging, and mitigation of charging artifacts. Implementation of Okapi-EM within the napari framework ensures that the tools are both user- and developer-friendly, through provision of a graphical user interface and access to Python programming.

A field experiment was conducted in 2019 and 2020 that included six site-years and four locations in Arkansas to determine the optimal sequence and timing of dicamba and glufosinate applications when applied alone, sequentially, or in combination to control Palmer amaranth by size: labeled (<10 cm height) and non-labeled (13 to 25 cm height). Single applications of dicamba, glufosinate, and dicamba plus glufosinate (not labeled) resulted in less than 80% Palmer amaranth control, regardless of weed size. The mixture of dicamba plus glufosinate was antagonistic for Palmer amaranth control and percent mortality. Sequential applications, averaged over all time intervals and herbicides, improved the percentage of Palmer amaranth control 11 to 17 percentage points over a single application, regardless of weed size at application 28 d after final application (DAFA). Palmer amaranth control with glufosinate followed by (fb) glufosinate and dicamba fb dicamba, pending weed size, were optimized at intervals of 7 d, and 14 to 21 d, respectively. Because single site of action (SOA) postemergence herbicide systems increase the likelihood of the development of resistant biotypes and are not a best management practice (BMP) in that regard; sequential applications involving both dicamba and glufosinate were more effective. Furthermore, the sequence of application mattered with a preference for applying dicamba first. Dicamba fb glufosinate at a 14-d interval was profit-maximizing and the only herbicide treatment that resulted in 100% weed control when size was <10 cm. For larger weed sizes, economic analysis revealed that dicamba fb dicamba performed better than dicamba fb glufosinate when no penalty was assigned for using a single SOA. This resulted in greater yield loss risk and soil weed seed bank in comparison to timelier weed control with the smaller weed size. Hence, timely weed control and two SOAs to control Palmer amaranth are recommended as BMPs that reduce producer risk.

Fluting is a technological and morphological hallmark of some of the most iconic North American Paleoindian stone points. Through decades of detailed artifact analyses and replication experiments, archaeologists have spent considerable effort reconstructing how flute removals were achieved, and they have explored possible explanations of why fluting was such an important aspect of early point technologies. However, the end of fluting has been less thoroughly researched. In southern North America, fluting is recognized as a diagnostic characteristic of Clovis points dating to approximately 13,000 cal yr BP, the earliest widespread use of fluting. One thousand years later, fluting occurs more variably in Dalton and is no longer useful as a diagnostic indicator. How did fluting change, and why did point makers eventually abandon fluting? In this article, we use traditional 2D measurements, geometric morphometric (GM) analysis of 3D models, and 2D GM of flute cross sections to compare Clovis and Dalton point flute and basal morphologies. The significant differences observed show that fluting in Clovis was highly standardized, suggesting that fluting may have functioned to improve projectile durability. Because Dalton points were used increasingly as knives and other types of tools, maximizing projectile functionality became less important. We propose that fluting in Dalton is a vestigial technological trait retained beyond its original functional usefulness.

From 2014 to 2020, we compiled radiocarbon ages from the lower 48 states, creating a database of more than 100,000 archaeological, geological, and paleontological ages that will be freely available to researchers through the Canadian Archaeological Radiocarbon Database. Here, we discuss the process used to compile ages, general characteristics of the database, and lessons learned from this exercise in “big data” compilation.

This study quantified CO2 emissions from tropical peat swamp soils in Brunei Darussalam. At each site, soil was collected from areas of intact and degraded peat and CO2 flux, and total organic content were measured ex situ. Soil organic content (~20–99%) was not significantly different between intact and degraded forest samples. CO2 flux was higher for intact forest samples than degraded forest samples (~1.0 vs. ~0.6 μmol CO2 m−2 s−1, respectively) but did not differ among forest locations. From our laboratory experiments, we estimated a potential emissions of ~10–20 t CO2 ha−1 y−1 which is in the lower range of values reported for other tropical peat swamps. However, our results are likely affected by unmeasured variation in root respiration and the lability of resident carbon. Overall, these findings provide experimental evidence to support that clearance of tropical peat swamp forests can increase CO2 emissions due to faster rates of decomposition.

We present the data and initial results from the first pilot survey of the Evolutionary Map of the Universe (EMU), observed at 944 MHz with the Australian Square Kilometre Array Pathfinder (ASKAP) telescope. The survey covers $270 \,\mathrm{deg}^2$ of an area covered by the Dark Energy Survey, reaching a depth of 25–30 $\mu\mathrm{Jy\ beam}^{-1}$ rms at a spatial resolution of $\sim$ 11–18 arcsec, resulting in a catalogue of $\sim$ 220 000 sources, of which $\sim$ 180 000 are single-component sources. Here we present the catalogue of single-component sources, together with (where available) optical and infrared cross-identifications, classifications, and redshifts. This survey explores a new region of parameter space compared to previous surveys. Specifically, the EMU Pilot Survey has a high density of sources, and also a high sensitivity to low surface brightness emission. These properties result in the detection of types of sources that were rarely seen in or absent from previous surveys. We present some of these new results here.