Objectives/Goals: Novel therapeutics to control Staphylococcus aureus (S. aureus) infections are needed for people with cystic fibrosis (CF, PwCF). In this study, our objective is to determine if the pharmacologic MEK1/2 inhibitor compound ATR-002 can restrict the growth of S. aureus clinical isolates and modulate infection in a murine model of S. aureus infection. Methods/Study Population: To evaluate the anti-inflammatory effects of ATR-002 on human macrophages, cells were stimulated with TLR2 agonists FSL1 or Pam3CSK4 with a dose range of ATR-002, and secretion of proinflammatory cytokines were measured by ELISA. To determine the direct antibacterial effect of ATR-002, minimum inhibitory concentration (MIC) assays were performed with the community-associated methicillin-resistant S. aureus strain USA300 and 40 S. aureus isolates from PwCF. To validate our results in vivo, mice were provided i.p. treatment with either vehicle, the MEK1/2 inhibitor compound PD0325901 (20 mg/kg), or ATR-002 (10 mg/kg) prior to intranasal infection with 1x10^7 CFU of USA300. Bacterial burdens at 4- and 24-hour post-infection (p.i.) and inflammatory cell recruitment at 24 hours p.i. were quantified. Results/Anticipated Results: Macrophages treated with ATR-002 exhibited a dose-dependent decrease in secretion of proinflammatory cytokines TNF and IL-8 following TLR stimulation. Our studies identified that ATR-002, but not PD0325901 or other MEK1/2 inhibitors, had direct antibacterial effects, and ATR-002 had an MIC range of 8 to above 64 ug/mL on CF S. aureus isolates. In the murine pulmonary infection model, delivery of ATR-002 and PD0325901 significantly prevented infection-induced loss of body mass and decreased neutrophil inflammation. However, when bacterial burdens were quantified 4-hours p.i., only ATR-002 treatment reduced lung bacterial burden compared to vehicle or PD0325901-treated groups. Discussion/Significance of Impact: These results are the first demonstration of the in vivo anti-inflammatory and antibacterial effects of ATR-002. Our results further demonstrate that ATR-002 exhibits direct antibacterial effects across a collection of clinical isolates of S. aureus. Future studies will continue to investigate the therapeutic potential of ATR-002.

Objectives/Goals: The never in mitosis kinase (NEK) family regulates vital processes, namely cell cycle progression, but their potential as therapeutic targets in TNBC has not been fully explored. Our studies aim to develop a toolkit to investigate the functional roles of NEKs in pathologies including carcinogenesis. Methods/Study Population: To assess differential NEK expression in normal and tumor tissues and correlation of gene expression with patient survival, we used Gene Expression Profiling Interactive Analysis (GEPIA) and Kaplan–Meier Plotter (KMPlot) pan-cancer analysis, respectively. Basal NEK protein levels were determined by immunoblot across a panel of cell lines, including breast cancer, osteosarcoma, hepatocellular carcinoma, and non-cancerous cells, to identify appropriate systems for evaluation of NEK function. Doxycycline-inducible cell lines were generated by transduction with lentiviral stocks of NEK shRNA and overexpression constructs and antibiotic selection. Expression was analyzed by qPCR and immunoblot. Results/Anticipated Results: Expression of NEK2, 4, 5, 6, 8, and 11 was higher in breast tumors compared to normal tissue by GEPIA analysis. Further examination using KMPlot showed a correlation between elevated NEK6 expression and decreased overall survival in patients with aggressive cancers. As an initial proof-of-concept study, we analyzed NEK6 protein expression in breast cancer cells. Levels of NEK6 were elevated in TNBC cells (MDA-MB-231) compared to hormone receptor positive (HR+) breast cancer cells (MCF7). Using complementary approaches to investigate the functional role of NEK6 in breast cancer, we depleted NEK6 expression using shRNAs in TNBC cells and expressed NEK6 in HR+ cells Discussion/Significance of Impact: Because kinase dysregulation promotes oncogenesis and metastasis, targeting kinases is a key strategy in therapeutic development. A NEK-specific molecular toolkit allows researchers to elucidate NEK functions and contributions to carcinogenesis, promoting advancement of novel therapies.

Insights into the paleoneurology and endocranial anatomy of ornithopod dinosaurs come largely from Northern Hemisphere taxa. The recently described non-hadrosaurid iguanodontian Fostoria dhimbangunmal from the Cenomanian of eastern Australia includes a partial skull that offers novel insights into its endocranial anatomy (i.e., the cavity housing the brain). Here, we describe the paleoneurology of F. dhimbangunmal based on a digital cranial endocast obtained from computed tomography. The endocast is mostly complete; however, it is diagenetically dorsoventrally compressed and its ventral limits are not preserved. The endocranial anatomy of F. dhimbangunmal is generally consistent with that of other non-hadrosaurid iguanodontians, including a well-developed olfactory apparatus, suggesting a good sense of smell. In contrast to hadrosaurids and some non-hadrosaurid iguanodontians, F. dhimbangunmal possesses the ancestral flexure condition, in which cranial and pontine flexure angles are subequal. The cerebrum makes up a significant portion of the endocast volume; however, the cerebral hemispheres are not as enlarged or bulbous as seen in hadrosaurids. The forebrain of F. dhimbangunmal did not fill the braincase to the same extent as in hadrosaurids. A distinct vacuity in the supraoccipital of F. dhimbangunmal may represent a new autapomorphy. This study provides the first insights into the neuroanatomy of an Australian iguanodontian dinosaur.

We use a combination of theory and experiment to study the incentives for firms to share knowledge when they engage in research and development (R&D) in an uncertain environment. We consider both symmetric and asymmetric starting points with regards to the amount of initial knowledge firms have before conducting R&D and look at how differences in starting positions affect the willingness of firms to share knowledge. We investigate when and if firms find R&D cooperation beneficial and how investment in R&D is affected by the outcome of the sharing decisions. The experimental evidence shows that overall subjects tend to behave consistently with theoretical predictions for the sharing of knowledge, although leaders who are not compensated by a side payment from laggards are more willing to share than predicted by the theory, and leaders who are compensated are less willing. The data on investment suggests less investment with sharing than without, consistent with theory. Compared to exact numerical predictions, there is overinvestment or underinvestment except for symmetric firms under no sharing. All cases of overinvestment and underinvestment, regardless of sharing or not and regardless of starting positions, are well explained by smoothed-out best (quantal) responses.

The recommended first-line treatment for insomnia is cognitive behavioral therapy for insomnia (CBTi), but access is limited. Telehealth- or internet-delivered CBTi are alternative ways to increase access. To date, these intervention modalities have never been compared within a single study. Further, few studies have examined a) predictors of response to the different modalities, b) whether successfully treating insomnia can result in improvement of health-related biomarkers, and c) mechanisms of change in CBTi. This protocol was designed to compare the three CBTi modalities to each other and a waitlist control for adults aged 50-65 years (N = 100). Participants are randomly assigned to one of four study arms: in-person- (n=30), telehealth- (n=30) internet-delivered (n=30) CBTi, or 12-week waitlist control (n=10). Outcomes include self-reported insomnia symptom severity, polysomnography, circadian rhythms of activity and core body temperature, blood- and sweat-based biomarkers, cognitive functioning, and magnetic resonance imaging.

We examined the association between influenza vaccination policies at acute care hospitals and influenza vaccination coverage among healthcare personnel for the 2021–22 influenza season. Mandatory vaccination and masking for unvaccinated personnel were associated with increased odds of vaccination. Hospital employees had higher vaccination coverage than licensed independent practitioners.

Perhaps because scholars of political ambition have focused almost entirely on electoral ambition, the presence of elections has been thought to play a major role in shaping who expresses interest in public service. In this article, we examine whether the presence or absence of elections changes women’s political ambition. Using surveys of law students, federal bureaucrats, and the general public, we find the relationship between gender and ambition for elected office is similar to the relationship between gender and ambition for bureaucratic and judicial service. We show that, although women are deterred from public service by the elections that act as gateways to those opportunities, the effects of elections on gendered political ambition duplicate the effects of other components of public service. Rather than unique, elections are duplicative in their effects, reinforcing the relationship between gender and ambition rather than fundamentally changing who expresses ambition for public service.

Why do US voters allow politicians to hold the country’s economy hostage during debt ceiling negotiations? In this research note, we argue that ignorance and uncertainty over the consequences of a debt ceiling breach play a nontrivial role in public support for hard-line negotiating positions. In a pre-registered survey experiment, two weeks before the June 2023 deadline to raise the US debt ceiling, we show that providing credible information about the consequences of default increases support for concessions among both Democrats and Republicans. Further, more certain information about the consequences of a debt ceiling breach has a larger effect than less-certain information suggesting that the unpredictable consequences of the crisis also help explain voter reluctance to accept concessions. The findings have implications for understanding debt ceiling negotiations and other crisis bargaining situations where the public serves as a relevant third party.

In humid, continental Michigan, we identified pedogenic carbonate in a soil profile developed on glacial drift sediments, as rinds, rhizoliths, and filaments (at depths >50 cm). Given that the climate setting is unusual for pedogenic carbonate, we investigated its formation with environmental monitoring and isotope analyses of carbonate (δ13C, δ18O, Δ47, and 14C) and waters (δ18O and δ2H). We found covariation in δ13C and Δ47 amongst the carbonate types (rhizoliths, rinds, filaments, bulk soil, and detrital clasts), and 14C ages of rinds that predate plausible formation ages. The δ13C and Δ47 values of the bulk carbonate and some of the pedogenic morphologies are not fully compatible with pedogenic formation in the modern environment. The δ18O data from precipitation and river waters and from carbonates are not uniquely identifying; they are compatible with the soil carbonate being pedogenic, detrital, or a mix. We conclude that the soil carbonate is likely a physical mix of pedogenic and detrital carbonate. Pedogenic carbonate is forming in this humid setting, likely because seasonal cycles in soil respiration and temperature cause cycles of dissolution and re-precipitation of detrital and pedogenic carbonate. The pedogenic carbonate may be a transient feature as carbonate-rich till undergoes post-glacial chemical weathering.

We have conducted a widefield, wideband, snapshot survey using the Australian SKA Pathfinder (ASKAP) referred to as the Rapid ASKAP Continuum Survey (RACS). RACS covers $\approx 90$% of the sky, with multiple observing epochs in three frequency bands sampling the ASKAP frequency range of 700–1 800 MHz. This paper describes the third major epoch at 1 655.5 MHz, RACS-high, and the subsequent imaging and catalogue data release. The RACS-high observations at 1 655.5 MHz are otherwise similar to the previously released RACS-mid (at 1 367.5 MHz) and were calibrated and imaged with minimal changes. From the 1 493 images covering the sky up to declination $\approx +48^\circ$, we present a catalogue of 2 677 509 radio sources. The catalogue is constructed from images with a median root-mean-square noise of $\approx 195$ $\unicode{x03BC}$Jy PSF$^{-1}$ (point-spread function) and a median angular resolution of $11{\stackrel{\prime\prime}{\raise-0pt\hbox{.}}}8 \times 8{\stackrel{\prime\prime}{\raise-0pt\hbox{.}}}1$. The overall reliability of the catalogue is estimated to be 99.18%, and we find a decrease in reliability as angular resolution improves. We estimate the brightness scale to be accurate to 10%, and the astrometric accuracy to be within $\approx 0{\stackrel{\prime\prime}{\raise-0pt\hbox{.}}}6$ in right ascension and $\approx 0{\stackrel{\prime\prime}{\raise-0pt\hbox{.}}}7$ in declination after correction of a systematic declination-dependent offset. All data products from RACS-high, including calibrated visibility datasets, images from individual observations, full-sensitivity mosaics, and the all-sky catalogue are available at the CSIRO ASKAP Science Data Archive.

The authors' proposal for the evolutionary origins of historical myths does not hold up to scrutiny, as illustrated by a simple mathematical model. Group-level explanations, such as defining the conditions for in-group membership, are dismissed by the authors but are far more plausible, as illustrated by the ongoing war in Ukraine.

We evaluated herbicides for controlling the annual grass ventenata [Ventenata dubia (Leers) Coss.], with particular interest in indaziflam, a preemergence cellulose biosynthesis inhibitor. In 2016, indaziflam was applied postemergence alone and in mixture with glyphosate, imazapic, propoxycarbazone-sodium, or rimsulfuron to an improved pasture in southwestern Montana. A non-sprayed control was included for comparison purposes. Canopy cover of each species was assessed annually for 7 yr; cover was grouped by life-form and longevity, and species richness was calculated. Five years (2021) after treatment, the seedbank was assessed. Our results indicated that treatments including indaziflam reduced V. dubia cover 1 to 3 yr and even up to 6 yr after application, with V. dubia cover being zero or close to zero. However, at 7 yr (2023) after treatment, V. dubia was low across all treatments, including the non-sprayed control. Perennial grasses and forbs and annual forbs were generally unaffected by any treatment and did not increase in cover over the 7 yr, even though V. dubia decreased. Two years after treatment, species richness was lowest in treatments that included indaziflam, but at 7 yr, species richness was similar across all treatments. Indaziflam depleted the monocot and dicot seedbank, with fewer than 5 seedlings of any species emerging from treatments that included indaziflam, while other treatments resulted in 60 to 165 seedlings per sample (40 cm3 of soil). In summary, at our study site, a single application of indaziflam controlled V. dubia for 6 yr, appeared to deplete the seedbank at 5 yr, and cover of perennial and annual vegetation and species richness was unaffected. By the end of the study, though, V. dubia cover appeared to be influenced by factors other than herbicide treatments, possibly variable precipitation over time, an exclusion of grazing, and competitive perennial grasses dominating the site.

Non-native plants negatively impact ecosystems via a variety of mechanisms, including in forested riparian areas. Japanese knotweed [Polygonum cuspidatum Siebold & Zucc.] and its hybrids (referred to as Polygonum spp. hereafter) are widely spread throughout North America and can impact flora and fauna of riparian habitats. Thus, information improving our ability to understand and predict the potential spread and colonization of Polygonum spp. is valuable. One dispersal mechanism is hydrochory (i.e., dispersal by water), including the downstream dispersal of viable stems that can facilitate rapid invasion within a watershed. We used passive integrated transponder (PIT) telemetry in experimental releases of Polygonum spp. stems to track the downstream transport of Polygonum spp. in a small (second-order) stream in northern New Hampshire, USA, in the summers of 2021 and 2022. A total of 180 (90 each year) Polygonum spp. stems were released at three sites within the stream reach, with 185 (∼98%) being recaptured at least once, with a total of 686 recaptures. Individual relocated stems moved a maximum distance of 30 to 875 m downstream in 2021 and 13 to 1,233 m in 2022 during regular flows; however, a high-streamflow event in July 2021 flushed out all remaining stems downstream of the study area. Generalized additive mixed models (GAMMs) identified site-specific differences in stem movement rates and a general reduction in movement rates with increased duration of time elapsed since post-release. In general, Polygonum spp. stems moved farther downstream in sites with lower channel sinuosity, although other fine-scale habitat factors (e.g., water depth, habitat type, and presence of wood and debris jams) likely contribute to the ability for Polygonum spp. to further disperse or otherwise be retained within the channel. Thus, stream morphology and stream flow are likely to affect where Polygonum spp. stems will be retained and potentially reestablish. Predictive tools identifying areas of higher probability of hydrochory-based dispersal could help to focus removal efforts when employed or to identify riparian habitats at highest risk for spread.