The COVID-19 pandemic highlighted gaps in infection control knowledge and practice across health settings nationwide. The Centers for Disease Control and Prevention, with funding through the American Rescue Plan, developed Project Firstline. Project Firstline is a national collaborative aiming to reach all aspects of the health care frontline. The American Medical Association recruited eight physicians and one medical student to join their director of infectious diseases to develop educational programs targeting knowledge gaps. They have identified 5 critical areas requiring national attention.

Effusion cooling is the state-of-the-art cooling technology for gas turbine hot-gas path components. Typically, effusion cooling holes across the entire combustor liner are aligned with the combustor axis, rendering a nominal zero compound angle between highly directional miniature effusion cooling jets and the main flow direction. The pitch of effusion cooling holes is optimised accordingly. However, the swirling main flow results in a non-zero compound angle and an effectively different pitch from the design. The directional effect of effusion cooling as a result of swirling main flow on the adiabatic film cooling effectiveness (AFE) is a combined effect of a non-zero compound angle and a varied pitch. The current experimental study aims to investigate the isolated effects of compound angle on AFE by excluding the influences of varying pitch. With an improved understanding of the sole effects of non-zero compound angles on AFE, the roles that a varied pitch plays in modifying AFE are further discussed to guide future effusion cooling designs under swirling main flow conditions. Binary pressure sensitive paint (PSP) was used to determine AFE experimentally.

We investigated experimentally the settling behaviour of vertically aligned spherical particles within various quiescent media at different release frequencies. The particles had a diameter of $d = 4$ mm and density of $\rho _s = 2200$ kg m$^{-3}$, and were released near the free surface of water, ethanol, a G60 water–glycerine mixture (60 % glycerine by weight) and oil media at frequencies of $f_P = 4$, 6 and 8 Hz, thereby allowing study of Galileo numbers, $Ga \in [16, 976]$. Particle tracking velocimetry quantified the motion of nearly 800 particles in a 600 mm high tank, and particle image velocimetry examined flow patterns around the particles. Results revealed that the centre of mass of the particle trajectories exhibited preferential in-plane motions, with significant lateral dispersion and large $Ga$ in water and ethanol, and nearly vertical paths with low $Ga$ in the G60 mixture and oil media. Varying degrees of particle separation resulted in higher terminal velocities than for a single particle. Hence, particle drag decreased in all cases, with the oil medium showing the highest drag reduction under the closest particle separation, reaching up to nearly 70 % of that for the single particle. The vertical and lateral pair dispersions, $R^2_z$ and $R^2_L$, exhibited ballistic scaling, with dependences on the initial separation, $r_0$, and the type of medium. With large $Ga$, $R^2_z$ displayed a ballistic regime followed by a slower rate, whereas with small $Ga$, $R^2_z$ maintained a consistent ballistic regime throughout settling. Finally, normalized $R^2_z$ demonstrated distinct scaling (exponent 2/3 and 1) dependent on the normalized initial separation and $Ga$.

We experimentally explored the effect of single-sidewall cooling on Rayleigh–Bénard (RB) convection. Canonical RB was also studied to aid insight. The scenarios shared tank dimensions and bottom and top wall temperatures; the single sidewall cooling had the top wall temperature. Turbulence was explored at two canonical Rayleigh numbers, $Ra=1.6\times 10^{10}$ and $Ra=2\times 10^9$ under Prandtl number $Pr=5.4$. Particle image velocimetry described vertical planes parallel and perpendicular to the sidewall cooling. The two $Ra$ scenarios reveal pronounced changes in the flow structure and large-scale circulation (LSC) due to the sidewall cooling. The density gradient induced by the sidewall cooling led to asymmetric descending and ascending flows and irregular LSC. Flow statistics departed from the canonical case, exhibiting lower buoyancy effects, represented by an effective Rayleigh number with effective height dependent on the distance from the lateral cooling. Velocity spectra show two scalings, $\varPhi \propto f^{-5/3}$ Kolmogorov (KO41) and $\varPhi \propto f^{-11/5}$ Bolgiano (BO59) in the larger $Ra$; the latter was not present in the smaller set-up. The BO59 scaling with sidewall cooling appears at higher frequencies than its canonical counterpart, suggesting weaker buoyancy effects. The LSC core motions allowed us to identify a characteristic time scale of the order of vortex turnover time associated with distinct vortex modes. The velocity spectra of the vortex core oscillation along its principal axis showed a scaling of $\varPhi _c \propto f^{-5/3}$ for the single sidewall cooling, which was dominant closer there. It did not occur in the canonical case, evidencing the modulation of LSC oscillation on the flow.

The distinct turbulence dynamics and transport modulated by a common seagrass species were investigated experimentally using a flexible surrogate canopy in a refractive-index-matching environment that enabled full optical access. The surrogate seagrass replicated the dynamic behaviour and morphological properties of its natural counterpart. The flows studied were subcritical with Froude numbers $Fr<0.26$ and concerned five Reynolds numbers $Re\in [3.4\times 10^{4}, 1.1\times 10^{5}]$ and Cauchy numbers $Ca\in [120, 1200]$. Complementary rigid canopy experiments were also included to aid comparative insight. The flow was quantified in wall-normal planes in a developed region using high-frame-rate particle image velocimetry. Results show that the deflection and coordinated waving motion of the blades redistributed the Reynolds stresses above and below the canopy top. Critically, in-canopy turbulence associated with the seagrass lacked periodic stem wake vortex shedding present in the rigid canopy, yet the flexible canopy induced vortex shedding from the blade tips. Inspection of spatial and temporal characteristics of coherent flow structures using spectral proper orthogonal decomposition reveals that Kelvin–Helmholtz-type vortices are the dominant flow structures associated with the waving motion of the seagrass and that modulated the local flow exchange in both rigid and flexible canopies. A barrier-like effect produced by the blade deflections blocked large-scale turbulence transport, thereby reducing vortex penetration into the canopy. In addition, we uncovered a transition from sweep-dominated to ejection-dominated behaviour in the surrogate seagrass. We hypothesise that the vortices created during the upward blade motion period play a major role in the sweep-to-ejection-dominated transition. Conditionally averaged quadrant analysis on the downward and upward blade motion supports this contention.

ABSTRACT IMPACT: Cholecystokinin-B Receptor -Mediates Growth of Hepatocellular Carcinoma with the use proglumide. Proglumide is a non-selective antagonistic drug therefore, strategies that block signaling at the CCK-BR may provide to be a novel therapeutic option for Hepatocellular Carcinoma treatment OBJECTIVES/GOALS: Cholecystokinin (CCK)and gastrin mediate the growth of Hepatocellular Carcinoma (HCC) through CCK-R and interruption of this signaling pathway could decrease HCC. CCK-Receptors are overexpressed in HCC and proliferation may be mediated through CCK-B. Blockade of the CCK-BR with proglumide decreased both growth in vitro and tumor growth in vivo. METHODS/STUDY POPULATION: RNA was extracted from murine Hepa1-6, RIL-175 and human HepG2 cells and was evaluated by qRT-PCR for expression of CCK-AR, CCK-BR and gastrin. CCK-R protein expression was analyzed by flow cytometry. HCC cells were treated in vitro with CCK peptide, the CCK-AR antagonist or the CCK-BR antagonist. Proliferation of selective CCK-R KO cells was compared to that of wild-type cells. To determine the effect of a CCK-R antagonist on tumor growth in vivo two cohorts of mice bearing subcutaneous Hepa1-6 or RIL-175 HCC tumors were treated with an oral bioavailable CCK-R antagonist proglumide or untreated water for 3-4 weeks. The mice bearing Hepa1-6 tumors were placed on a high-fat diet to raise blood CCK levels. Mice bearing RIL-175 tumors were fed standard chow to determine if proglumide could block autocrine growth by gastrin. RESULTS/ANTICIPATED RESULTS: The mRNA expression of CCK-AR, CCK-BR and gastrin were increased 80-90-fold in all HCC cell lines compared to that of normal liver. CCK-BRs were detected on >85% of the cells by flow cytometry. CCK peptide (1nM) stimulated HCC growth in vitro in both wild-type cells and in CCK-AR KO cells but not in CCK-BR KO cells. CCK-BR antagonist blocked CCK-stimulated growth in vitro but the CCK-AR antagonist did not, suggesting that the CCK-BR was responsible for mediating proliferation. In vivo tumor growth was significantly reduced with proglumide treatment by 70% (p<0.05) in Hepa1-6 and by 73% (p<0.001) in RIL-75 tumors, respectively. DISCUSSION/SIGNIFICANCE OF FINDINGS: CCK-Rs are overexpressed in HCC and proliferation appears to be mediated through the CCK-BR. Downregulation with CRISPR Cas9 or blockade of the CCK-BR with an antagonist decreases growth in vitro and proglumide therapy decreases tumor growth in vivo. Strategies that block signaling at the CCK-BR maybe a novel therapeutic option for HCC treatment.

Ex situ conservation of species is risky and expensive, but it can prevent extinction when in situ conservation fails. We used the IUCN Guidelines on the Use of Ex Situ Management for Species Conservation to evaluate whether to begin ex situ conservation for the South-east Asian subspecies of Bengal florican Houbaropsis bengalensis blandini, which is predicted to be extinct in the wild within 5 years. To inform our decision, we developed a decision tree, and used a demographic model to evaluate the probability of establishing a captive population under a range of husbandry scenarios and egg harvest regimes, and compared this with the probability of the wild population persisting. The model showed that if ex situ conservation draws on international best practice in bustard husbandry there is a high probability of establishing a captive population, but the wild population is unlikely to persist. We identified and evaluated the practical risks associated with ex situ conservation, and documented our plans to mitigate them. Modelling shows that it is unlikely that birds could be released within 20–30 years, by which time genetic, morphological and behavioural changes in the captive population, combined with habitat loss and extinction of the wild population, make it unlikely that Bengal florican could be released into a situation approximating their current wild state. We considered the philosophical and practical implications through a decision tree so that our decision to begin ex situ management is not held back by our preconceived notions of what it means to be wild.

Laboratory experiments are conducted to investigate the mechanism controlling the formation of stable and unstable acoustic fountains at the free surface of a quiescent body of water. Fountains are induced by focused ultrasonic, a new modality that allows for better spatiotemporal control of water flow. Particle image velocimetry was used to characterize the induced flow field in the vicinity of the ultrasonic focal spot. We used two types of ultrasonic transducers with distinct wave frequencies. We examined three fountain formation regimes by varying the pressure level of the transducers, namely weak, intermediate (stable) and highly forced fountains (explosive). Between different regimes, the fountain height underwent a step-change in response to the increase in acoustic pressure. A force estimation obtained from the flow field shows that the magnitude of axial momentum flux is orders of magnitude lower than that of gravity and surface tension, indicating that the dominant driving force for the fountain generation is the acoustic radiation force (Nightingale et al., Ultrasound Med. Biol., vol. 28, 2002, pp. 227–235). We propose a simple model to estimate the shape of a stable fountain; it accounts for the applied acoustic pressure, gravity, surface tension and axial momentum. The model neglects viscous force, which precludes capturing the intermediate fountain surface curvature. However, the model successfully predicts the geometry of the weak and intermediate fountains.

Recently, innovations in ice drilling have yielded considerable improvements to existing drilling techniques, as well as innovative drilling technologies that can be used in new types of applications. However, some specific challenges have to be addressed for improving existing drilling methods and developing new ones: (1) combination and unification of different drilling systems; (2) facilitating ice core breaking; (3) improving existing systems and developing new rapid-access ice drilling systems; (4) reliable elimination of ice hydraulic fracturing problems; (5) developing new environment-friendly methods of drilling in the sub-glacial lake sediments; and (6) design of unconventional ice drilling systems. Possible solutions to these problems are presented herein.

The AD 775 peak in Δ14C (henceforth, M12) was first measured by Miyake et al. and has since been confirmed globally. Here we present earlywood and latewood Δ14C values from tree rings of pinyon pine (Pinus edulis) from Mummy Cave, Canyon de Chelly National Monument, Chinle, Arizona, USA, for the period AD 770–780. These data reconfirm the timing of M12 and show a small rise in Δ14C in AD 774 latewood. Allowing for the delay in lateral transfer of radiocarbon produced at high latitude, this suggests that 14C peak production occurred in late winter or spring of AD 774. Additionally, Δ14C decreased slightly in the earlywood of AD 775 and increased in the latewood of AD 775 to a higher level than that observed in AD 774.

Praziquantel (PZQ) is the drug of choice for schistosomiasis. The potential drug resistance necessitates the search for adjunct or alternative therapies to PZQ. Previous functional genomics has shown that RNAi inhibition of Ca2+/calmodulin-dependent protein kinase II (CaMKII) gene in Schistosoma adult worms significantly improved the effectiveness of PZQ. Here we tested the in vitro efficacy of 15 selective and non-selective CaMK inhibitors against Schistosoma mansoni and showed that PZQ efficacy was improved against refractory juvenile parasites when combined with these CaMK inhibitors. By measuring CaMK activity and the mobility of adult S. mansoni, we identified two non-selective CaMK inhibitors, Staurosporine (STSP) and 1Naphthyl PP1 (1NAPP1), as promising candidates for further study. The impact of STSP and 1NAPP1 was investigated in mice infected with S. mansoni in the presence or absence of a sub-lethal dose of PZQ against 2- and 7-day-old schistosomula and adults. Treatment with STSP/PZQ induced a significant (47–68%) liver egg burden reduction compared with mice treated with PZQ alone. The findings indicate that the combination of STSP and PZQ dosages significantly improved anti-schistosomal activity compared to PZQ alone, demonstrating the potential of selective and non-selective CaMK/kinase inhibitors as a combination therapy with PZQ in treating schistosomiasis.

Toxoplasma gondii rhoptry protein TgROP18 is a polymorphic virulence effector that targets immunity-related GTPases (IRGs) in rodents. Given that IRGs are uniquely diversified in rodents and not in other T. gondii intermediate hosts, the role of TgROP18 in manipulating non-rodent cells is unclear. Here we show that in human cells TgROP18I interacts with the interferon-gamma-inducible protein N-myc and STAT interactor (NMI) and that this is a property that is unique to the type I TgROP18 allele. Specifically, when expressed ectopically in mammalian cells only TgROP18I co-immunoprecipitates with NMI in IFN-γ-treated cells, while TgROP18II does not. In parasites expressing TgROP18I or TgROP18II, NMI only co-immunoprecipitates with TgROP18I and this is associated with allele-specific immunolocalization of NMI on the parasitophorous vacuolar membrane (PVM). We also found that TgROP18I reduces NMI association with IFN-γ-activated sequences (GAS) in the IRF1 gene promoter. Finally, we determined that polymorphisms in the C-terminal kinase domain of TgROP18I are required for allele-specific effects on NMI. Together, these data further define new host pathway targeted by TgROP18I and provide the first function driven by allelic differences in the highly polymorphic ROP18 locus.

OBJECTIVES/GOALS: Non-alcoholic steatohepatitis (NASH) is a leading cause of cirrhosis in the world for which no anti-fibrotic therapies exist. We hypothesized that BMS-22 and maraviroc (MVC), chemokine receptor 2 (CCR2) and 5 (CCR5) antagonists, respectively, would diminish the fibrogenic activity of "fat-exposed" murine pHSCs. METHODS/STUDY POPULATION: pHSCs were isolated from livers of 6 week old male mice following 4 weeks on a NASH-inducing choline-deficient high fat diet (CDAHFD, “fat-exposed”) or standard diet (SD) and passaged in vitro. Early passage (6-12) pHSCs were plate-adhered and TGF-b-treated (10ng/mL) to maximally activate their pro-fibrogenic genes, collagen 1α1 (Col1A1), tissue inhibitor of metalloproteinase 1 (TIMP1), or α-smooth muscle actin (ACTA2). CDAHFD and SD pHSCs were then treated for 48 hours with increasing doses of BMS-22 or MVC (range: 0.3-120ng/mL) to determine (1) the degree of attenuation of the pro-fibrogenic response as measured by qPCR of fibrogenic genes (Col1A1, TIMP1,ACTA2); (2) enhancement of a fibrolytic response as measured by qPCR of matrix metalloproteinases (MMP) 2, 9 and 13 genes; and (3) pHSC migration using the scratch assay. Cell viability and CCR2 and CCR5 gene expression in response to escalating doses of antagonists were also measured. RESULTS/ANTICIPATED RESULTS: Plate- and TGF-b activated CDAHFD pHSCs had a 2-fold greater, dose-dependent attenuation of their pro-fibrogenic activity in response to BMS-CCR2-22 and MVC, when compared with plate- and TGF-b activated SD pHSCs, as measured by reductions in collagen 1α1 (Col1A1) and α-smooth muscle actin (ACTA2) gene expression. TIMP1 gene expression was unaffected by drug treatment for 48 hours. Cell viability was not affected up to doses of 30ng/mL of each drug. pHSCs also demonstrated a dose-dependent increase in CCR2, CCR5 and MMP-9 gene expression in response to surface receptor antagonism. Migration assays comparing CDAHFD and SD pHSCs in response to escalating doses of MVC and BMS-22 are ongoing and expected to demonstrate a significantly decreased migratory capacity of CDAHFD pHSCs than SD pHSCs in response to therapy, reflecting the increased susceptibility of the “fat-exposed” pHSCs to anti-fibrotic therapy than normal pHSCs. DISCUSSION/SIGNIFICANCE OF IMPACT: Anti-fibrotic drugs that dampen pro-fibrogenic activities of “fat-exposed” pHSCs are urgently needed. CCR2 and CCR5 antagonists, BMS-22 and MVC, respectively, can selectively dampen the pro-fibrogenic response of fat-exposed pHSCs, and must be considered for future trials in human NASH. CONFLICT OF INTEREST DESCRIPTION: Dr. Jill Smith has a patent licensing agreement with Immune Therapeutics, Inc.