OBJECTIVES/GOALS: Neuroblastoma (NB) is an embryonal cancer of the sympathetic nervous system that affects mostly infants and young children. The complex genetic background present across NB patients results in diverse clinical response and difficulty in individualizing therapy. Currently, NB patients undergo a regimen of genotoxic chemotherapeutics, radiation therapy, and new immunotherapy that, while effective, has significant side effects, including excruciating pain. One promising avenue for targeted therapy in neuroblastoma focuses on anaplastic lymphoma kinase (ALK), a cell surface neural receptor tyrosine kinase. We previously identified activating point mutations within the tyrosine kinase domain of ALK as the primary cause of hereditary NB, and we and others subsequently showed that these same alterations are the most common somatic single-nucleotide mutations in the sporadic forms of the disease. Crizotinib, a first-generation small molecule ATP-competitive inhibitor of the ALK tyrosine kinase, showed limited anti-tumor activity in patients with relapsed NB harboring ALK F1174 and F1245 mutations. We have demonstrated that lorlatinib, a novel ATP-competitive ALK inhibitor, overcomes this de novo resistance in preclinical models of ALK-driven NB. Recent clinical trials with lorlatinib in patients with non-small cell lung cancer harboring an ALK fusion, and in patients with NB harboring ALK mutations show the emergence of multiple or compound ALK mutations as a mechanism of resistance. We postulate that these compound mutations disrupt the interaction between and ALK and cause resistance. In this study, we employ a computational approach to model mutated ALK in complex with lorlatinib as well as ATP to understand whether the new mutations alter the affinity or mode of lorlatinib/ATP binding to ALK, and thus cause suboptimal ALK inhibition. METHODS/STUDY POPULATION: We employ methods in computational structural biology and drug design, primarily based on molecular modeling, molecular dynamics (MD), and molecular docking. Based on existing crystal structures of wildtype ALK, we model the mutations and perform MD simulations in order to characterize the activation state of the protein as well as perform ensemble docking calculations to assess the binding affinities and modes in ALK-lorlatinib and ALK-ATP complexes. RESULTS/ANTICIPATED RESULTS: We expect that the compound mutations cause resistance to lorlatinib either by lowering protein affinity for the drug or increasing the affinity for ATP. Alternatively, the compound mutations may disrupt the protein activation state, in which case ALK may no longer be active, and another protein/pathway could be driving the resistance. DISCUSSION/SIGNIFICANCE OF IMPACT: The results of this study will enable the understanding of the mechanism of resistance to lorlatinib and facilitate the design of new ALK inhibitors, or help develop more optimal and mechanism-guided therapies aimed to overcome the resistance.

Restless legs syndrome (RLS) is a neurological disorder characterized by an urge to move and uncomfortable sensations. Genetic studies have identified polymorphisms in up to 19 risk loci, including MEIS1 and BTBD9. Rodents deficient in either homolog show RLS-like phenotypes. However, whether MEIS1 and BTBD9 interact in vivo is unclear. Here, with C. elegans, we observed that the hyperactive egg-laying behavior caused by loss of BTBD9 homolog was counteracted by knockdown of MEIS1 homolog. This was further investigated in mutant mice with Btbd9, Meis1, or both knocked out. The double knockout mice showed an earlier onset of the motor deficit in a wheel running test but did not have increased sensitivity to heat stimuli as observed in single knock outs. Meis1 protein level was not influenced by Btbd9 deficiency, and Btbd9 transcription was not affected by Meis1 haploinsufficiency. Our results demonstrate that MEIS1 and BTBD9 do not regulate each other.

Field trials were conducted in North Carolina in 2017 and Louisiana and Mississippi in 2018 to determine the effect of pretransplanting applications of diquat on sweetpotato crop tolerance, yield, and storage root quality. In North Carolina treatments consisted of two rates of diquat (560 or 1,120 g ai ha−1) alone or mixed with 107 g ai ha−1 flumioxazin and applied 1 d before transplanting (DBP), sequential applications of diquat (560 or 1,120 g ha−1) 1 and 17 DBP, 107 g ha−1 flumioxazin alone, and a nontreated check. In Louisiana and Mississippi treatments consisted of diquat (560 or 1,120 g ha−1) applied 1 DBP either alone or followed by (fb) rehipping rows or 107 g ha−1 flumioxazin immediately prior to transplanting. Additional treatments included 546 g ha−1 paraquat applied 1 DBP and a nontreated check. In North Carolina injury was ≤3% for all treatments through 23 d after transplanting (DAP), and no injury was observed after 23 DAP. Visual sweetpotato stunting pooled across the Mississippi and Louisiana trials ranged from 1% to 14%, 0% to 6%, and 0% to 3% at 2, 4, and 6 wk after planting (WAP), respectively, and no crop injury was observed after 6 WAP. Diquat applied 1 DBP and not fb rehipping resulted in greater crop injury (12%) than comparable treatments that were rehipped (2%). In North Carolina single and sequential diquat applications resulted in reduced No. 1 sweetpotato yield (24,230 and 24,280 kg ha−1, respectively) compared with the nontreated check, but No. 1 yield when diquat plus flumioxazin (26,330 kg ha−1) was used was similar to that of the nontreated check. No. 1 yield did not differ by treatment in Louisiana and Mississippi.

Field trials were conducted near Pontotoc, Mississippi; Chase, Louisiana; and Clinton, North Carolina, in 2017 and 2018 to determine the effect of pendimethalin rate and timing application on sweetpotato crop tolerance, yield, and storage root quality. Treatments consisted of five pendimethalin rates (266, 532, 1,065, 1,597, and 2,130 g ai ha−1) by two application timings (0 to 1 or 10 to 14 d after transplanting). Additionally, a nontreated check was included for comparison. Crop injury (stunting) was minimal (≤4%) through 6 wk after transplanting (WAP) and no injury was observed from 8 to 14 WAP, regardless of application timing or rate. The nontreated check yielded 6.6, 17.6, 5.5, and 32.1 × 103 kg ha−1 of canner, no. 1, jumbo, and total grades, respectively. Neither pendimethalin application timing nor rate influenced jumbo, no. 1, marketable, or total sweetpotato yield. Overall, these results indicate that pendimethalin will be a valuable addition to the toolkit of sweetpotato growers.

The Japanese Murrelet Synthliboramphus wumizusume is a rare, globally ‘Vulnerable’ seabird, endemic to Japan and South Korea. However, little is known of its at-sea distribution, habitat or threats. We conducted several years of at-sea surveys around Japan to model Japanese Murrelet density in relation to habitat parameters, and make spatial predictions to assess the adequacy of the current Japanese marine Important Bird and Biodiversity Area (IBA) network for the species. During a five-year period, 3,485 km of at-sea surveys recorded 3,161 Japanese Murrelets around four breeding locations. Maximum murrelet group size was 90 individuals with a mean group size of 2.9 ± 4.2 individuals. Models of Japanese Murrelet at-sea density around the two largest breeding locations predicted that almost all murrelets occur within 30 km of the breeding colony and most within 10 km. Murrelets were predicted closer to the colony in May than in April and closer to the colony at a neritic colony than at an offshore island colony. Additionally, murrelets breeding on an offshore island colony also commuted to mainland neritic habitat for foraging. The marine habitat used by Japanese Murrelets differed between each of the four surveyed colonies, however oceanographic variables offered little explanatory power in models. Models with colony, month and year generated four foraging radii (9–39 km wide) containing murrelet densities of > 0.5 birds/km2. Using these radii the Japanese marine IBA network was found to capture between 95% and 25% of Japanese Murrelet at-sea habitat while breeding and appears appropriately configured to protect near-colony murrelet distributions. Given the range of marine habitats that breeding murrelets inhabit, our simple models offer an applicable method for predicting to unsampled colonies and generating ecologically-informed seaward extension radii. However, data on colony populations and further at-sea surveys are necessary to refine models and improve predictions.

Laboratory experiments were performed on a geometrically scaled vertical-axis wind turbine model over an unprecedented range of Reynolds numbers, including and exceeding those of the full-scale turbine. The study was performed in the high-pressure environment of the Princeton High Reynolds number Test Facility (HRTF). Utilizing highly compressed air as the working fluid enabled extremely high Reynolds numbers while still maintaining dynamic similarity by matching the tip speed ratio (defined as the ratio of tip velocity to free stream, $\unicode[STIX]{x1D706}=\unicode[STIX]{x1D714}R/U$ ) and Mach number (defined at the turbine tip, $Ma=\unicode[STIX]{x1D714}R/a$ ). Preliminary comparisons are made with measurements from the full-scale field turbine. Peak power for both the field data and experiments resides around $\unicode[STIX]{x1D706}=1$ . In addition, a systematic investigation of trends with Reynolds number was performed in the laboratory, which revealed details about the asymptotic behaviour. It was shown that the parameter that characterizes invariance in the power coefficient was the Reynolds number based on blade chord conditions ( $Re_{c}$ ). The power coefficient reaches its asymptotic value when $Re_{c}>1.5\times 10^{6}$ , which is higher than what the field turbine experiences. The asymptotic power curve is found, which is invariant to further increases in Reynolds number.

On August 25, 2017, Hurricane Harvey made landfall near Corpus Christi, Texas. The ensuing unprecedented flooding throughout the Texas coastal region affected millions of individuals.1 The statewide response in Texas included the sheltering of thousands of individuals at considerable distances from their homes. The Dallas area established large-scale general population sheltering as the number of evacuees to the area began to amass. Historically, the Dallas area is one familiar with “mega-sheltering,” beginning with the response to Hurricane Katrina in 2005.2 Through continued efforts and development, the Dallas area had been readying a plan for the largest general population shelter in Texas. (Disaster Med Public Health Preparedness. 2019;13:33–37)

The seventh meeting of the World Congress of Pediatric Cardiology and Cardiac Surgery was an opportunity for healthcare professionals from around the world to meet and discuss current issues affecting patients with acquired and CHD. A dedicated anaesthesia track facilitated the exchange of ideas and fostered many new friendships. This review highlights the congenital cardiac anaesthesia track and the involvement of the Congenital Cardiac Anesthesia Society in the congress.