The importance of studying the radiocarbon content of dissolved inorganic carbon (DI14C) in the oceans has been recognized for decades. Starting with the GEOSECS program in the 1970s, 14C sampling has been a part of most global survey programs. Early results were used to study air-sea gas exchange while the more recent results are critical for helping calibrate ocean general circulation models used to study the effects of climate change. Here we summarize the major programs and discuss some of the important insights the results are starting to provide.

OBJECTIVES/GOALS: The goal of this study was to understand the impact of a high sodium diet on gene networks in the kidney that correlate with blood pressure in female primates, and translating findings to women. METHODS/STUDY POPULATION: Sodium-naïve female baboons (n=7) were fed a low-sodium (LS) diet for 6 weeks followed by a high sodium (HS) diet for 6 weeks. Sodium intake, serum 17 beta-estradiol, and ultrasound-guided kidney biopsies for RNA-Seq were collected at the end of each diet. Blood pressure was continuously measured for 64-hour periods throughout the study by implantable telemetry devices. Weighted gene coexpression network analysis was performed on RNA-Seq data to identify transcripts correlated with blood pressure on each diet. Network analysis was performed on transcripts highly correlated with BP, and in silico findings were validated by immunohistochemistry of kidney tissues. RESULTS/ANTICIPATED RESULTS: On the LS diet, Na+ intake and serum 17 beta-estradiol concentration correlated with BP. Cell type composition of renal biopsies was consistent among all animals for both diets. Kidney transcriptomes differed by diet; analysis by unbiased weighted gene co-expression network analysis revealed modules of genes correlated with BP on the HS diet. Network analysis of module genes showed causal networks linking hormone receptors, proliferation and differentiation, methylation, hypoxia, insulin and lipid regulation, and inflammation as regulators underlying variation in BP on the HS diet. Our results show variation in BP correlated with novel kidney gene networks with master regulators PPARG and MYC in female baboons on a HS diet. DISCUSSION/SIGNIFICANCE: Previous studies in primates to identify molecular networks dysregulated by HS diet focused on males. Current clinical guidelines do not offer sex-specific treatment plans for sodium sensitive hypertension. This study leveraged variation in BP as a first step to identify correlated kidney regulatory gene networks in female primates after a HS diet.

In the December 2021 issue of Cardiology in the Young, Hubrechts and colleagues, from Brussels and Leuven in Belgium, describe their experience in which the pulmonary veins were normally connected to the morphologically left atrium. By virtue of the presence of a shelf dividing the morphologically left atrium, however, the venous return was to the morphologically right atrium, with no evidence of formation of the superior interatrial fold, meaning that there was no obstruction of flow into the systemic venous circulation. The question posed by the Belgian authors is whether the shelf dividing the morphologically left atrium is a deviated primary atrial septum, as the arrangement has previously been interpreted. As they discuss, it is currently impossible to arbitrate this conundrum. In our commentary, we discuss the background to the dilemma. We point out that, as yet, it is not possible to code accurately this congenital cardiac malformation within The International Paediatric and Congenital Cardiac Code (IPCCC), nor within the newly produced 11th Revision of the International Classification of Diseases (ICD-11).

OBJECTIVES/GOALS: A functional precision medicine platform to identify therapeutic targets for a glioblastoma patient with Li Fraumeni syndrome was performed. Comparative transcriptomics identified druggable targets and patient derived organoids and a 3D-PREDICT drug screening assay was used to validate the pipeline and identify further therapeutic targets. METHODS/STUDY POPULATION: A comparative transcriptomics pipeline was used to identify druggable genes that are uniquely overexpressed in our patient of interest relative to a cancer compendium of 12,747 tumor RNA sequencing datasets including 200 GBMs. Mini-ring patient derived organoid-based drug viability assays were performed to validate the comparative transcriptomics data. Additionally, a spheroid-based drug screening assay (3D-PREDICT) was performed and used to identify further therapeutic targets. RESULTS/ANTICIPATED RESULTS: Using comparative transcriptomics STAT1 and STAT2 were found to be significantly overexpressed in our patient, indicating ruxolitinib, a Janus kinase 1 and 2 inhibitor, as a potential therapy. Druggable pathways predicted using comparative transcriptomics corresponded with ruxolitinib sensitivity in a panel of patient derived organoids screened with this compound. Cells from the LFS patient were among the most sensitive to ruxolitinib compared to patient-derived cells with lower STAT1 and STAT2 expression levels. Additionally, 3D-PREDICT screening identified the mTOR inhibitor everolimus as a potential candidate. These two targeted therapies were selected for our patient and resulted in radiographic disease stability. DISCUSSION/SIGNIFICANCE: This research illustrates the use of comparative transcriptomics to identify druggable pathways irrespective of actionable DNA mutations present. Our results are promising and serve to highlight the importance of functional precision medicine in tailoring treatment regimes to specific patients.

OBJECTIVES/GOALS: Low statistical power is a problem is many fields. We performed a systematic review to determine the median statistical power of studies of epilepsy surgery outcomes. METHODS/STUDY POPULATION: We performed a PubMed search for studies reporting epilepsy surgery outcomes for the years 1980-2000, focusing on studies using stereo-electroencephalography (SEEG). We extracted patient count data for comparisons of surgical outcome between groups, based on a prognostic factor. We defined a clinically meaningful difference the surgical outcome for MRI positive (66.9%) compared to MRI negative (45.5%) in the largest study in the series. The statistical power of a Chi-square test was computed as the percentage of simulated runs (10,000 repetitions) assuming this difference with a p-value less than 0.05. RESULTS/ANTICIPATED RESULTS: Based on 69 studies, the median sample size was 38 patients, and the median statistical power was 24%. This implies at least a 17% (0.5/[0.24+0.05)) chance a study with a significant result in false, assuming 1:1 pre-test odds. A 'typical’ SEEG study with 33 patients and 2:1 allocation had a median significant odds ratio of 6.5, which over-estimates the true odds ratio of 2.4. DISCUSSION/SIGNIFICANCE: Studies of epilepsy surgery outcomes using SEEG are statistically underpowered. This means true effects will be missed, the chance a study with a significant result is false will be inflated, and significant effects found will be over-estimated. Studies of surgery outcome need better statistical rigor if they are to reliably guide treatment.

OBJECTIVES/GOALS: Recent research has attempted to identify diagnostic, prognostic, and predictive biomarkers, however, currently, no biomarkers can accurately diagnose GBC and predict patients prognosis. Using machine learning, we can utilize high-throughput RNA sequencing with clinicopathologic data to develop a predictive tool for GBC prognosis. METHODS/STUDY POPULATION: Current predictive models for GBC outcomes often utilize clinical data only. We aim to build a superior algorithm to predict overall survival in GBC patients with advanced disease, using machine learning approaches to prioritize biomarkers for GBC prognosis. We have identified over 80 fresh frozen GBC tissue samples from Rochester, Minnesota, Daegu, Korea, Vilnius, Lithuania, and Calgary, Canada. We will perform next-generation RNA sequencing on these tissue samples. The patients clinical, pathologic and survival data will be abstracted from the medical record. Random forests, support vector machines, and gradient boosting machines will be applied to train the data. Standard 5-fold cross validation will be used to assess performance of each ML algorithm. RESULTS/ANTICIPATED RESULTS: Our preliminary analysis of next generation RNA sequencing from 18 GBC tissue samples identified recurrent mutations in genes enriched in pathways in cytoskeletal signaling, cell organization, cell movement, extracellular matrix interaction, growth, and proliferation. The top three most significantly altered pathways, actin cytoskeleton signaling, hepatic fibrosis/hepatic stellate cell activation, and epithelial adherens junction signaling, emphasized a molecular metastatic and invasive fingerprint in our patient cohort. This molecular fingerprint is consistent with the previous knowledge of the highly metastatic nature of gallbladder tumors and is also manifested physiologically in the patient cohort. DISCUSSION/SIGNIFICANCE: Integrative analysis of molecular and clinical characterization of GBC has not been fully established, and minimal improvement has been made to the survival of these patients. If overall survival can be better predicted, we can gain a greater understanding of key biomarkers driving the tumor phenotype.

We honour a great man and a true giant. Lodewyk H.S. van Mierop (March 31, 1927 – October 17, 2021), known as Bob, was not only a Paediatric Cardiologist but also a dedicated Scientist. He made many significant and ground-breaking contributions to the fields of cardiac anatomy and embryology. He was devoted as a teacher, spending many hours with medical students, Residents, and Fellows, all of whom appreciated his regularly scheduled educational sessions. Those of us who were fortunate to know and spend time with him will always remember his great mind, his willingness to share his knowledge, and his ability to encourage spirited and fruitful discussions. His life was most productive, and he will long be remembered by many through his awesome and exemplary scientific contributions.

His legacy continues to influence the current and future generations of surgeons and all providers of paediatric and congenital cardiac care through the invaluable archive he established at University of Florida in Gainesville: The University of Florida van Mierop Heart Archive. Undoubtedly, with these extraordinary contributions to the fields of cardiac anatomy and embryology, which were way ahead of his time, Professor van Mierop was a true giant in Paediatric Cardiology. The invaluable archive he established at University of Florida in Gainesville, The University of Florida van Mierop Heart Archive, has been instrumental in teaching medical students, Residents, Medical Fellows, and Surgical Fellows. Only a handful of similar archives exist across the globe, and these archives are the true legacy of giants such as Dr. van Mierop. We have an important obligation to leave no stone unturned to continue to preserve these archives for the future generations of surgeons, physicians, all providers of paediatric and congenital cardiac care, and, most importantly, our patients.

Self-control failure occurs when an individual experiences a conflict between immediate desires and longer-term goals, recognises psychological forces that hinder goal-directed action, tries to resist them but fails in the attempt. Behavioural economists often invoke assumptions about self-control failure to justify proposals for policy interventions. These arguments require workable methods for eliciting individuals’ goals and for verifying occurrences of self-control failure, but developing such methods confronts two problems. First, it is not clear that individuals’ goals are context-independent. Second, facing an actual conflict between a desire and a self-acknowledged goal, a person may consciously choose not to resist the desire, thinking that spontaneity is more important than self-control. We address these issues through an online survey that elicited individuals’ self-reported judgements about the relative importance of self-control and spontaneity in conflicts between enjoyment and health-related goals. To test for context-sensitivity, the judgement-elicitation questions were preceded by a memory recall task which directed participants’ attention either to the enjoyment of acting on desires or to the satisfaction of achieving goals. We found little evidence of context-sensitivity. In both treatments, however, judgements that favoured spontaneity were expressed with roughly the same frequency and strength as judgments that favoured self-control.

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.