In The Pune Maternal Nutrition Study, vitamin B12 deficiency was seen in 65% of pregnant women, folate deficiency was rare. Maternal total homocysteine concentrations were inversely associated with offspring birthweight, and low vitamin B12 and high folate concentrations predicted higher offspring adiposity and insulin resistance. These findings guided a nested pre-conceptional randomised controlled trial ‘Pune Rural Intervention in Young Adolescents’. The interventions included: (1) vitamin B12+multi-micronutrients as per the United Nations International Multiple Micronutrient Antenatal Preparation, and proteins (B12+MMN), (2) vitamin B12 (B12 alone), and (3) placebo. Intervention improved maternal pre-conceptional and in-pregnancy micronutrient nutrition. Gene expression analysis in cord blood mononuclear cells in 88 pregnancies revealed 75 differentially expressed genes between the B12+MMN and placebo groups. The enriched biological processes included G2/M phase transition, chromosome segregation, and nuclear division. Enriched pathways included, mitotic spindle checkpoint and DNA damage response while enriched human phenotypes were sloping forehead and decreased head circumference. Fructose-bisphosphatase 2 (FBP2) and Cell Division Cycle Associated 2 (CDCA2) genes were under-expressed in the B12 alone group. The latter, involved in chromosome segregation was under-expressed in both intervention groups. Based on the role of B-complex vitamins in the synthesis of nucleotides and S-adenosyl methionine, and the roles of vitamins A and D on gene expression, we propose that the multi-micronutrient intervention epigenetically affected cell cycle dynamics. Neonates in the B12+MMN group had the highest ponderal index. Follow-up studies will reveal if the intervention and the altered biological processes influence offspring diabesity.

Politiko-Troullia has generated the largest radiocarbon (14C) dataset from a Prehistoric Bronze Age settlement on Cyprus. We present Bayesian modeling of 25 calibrated AMS ages, which contributes to an emerging multi-site 14C chronology for Cyprus covering most of the Prehistoric Bronze Age. Our analysis places the six stratified phases of occupation at Troullia between about 2050 and 1850 cal BCE, in contrast to a longer estimated occupation inferred from pottery analysis. We provide a rare 14C determination for the transition from Prehistoric Bronze Age 1 to 2 just after 2000 cal BCE, associated with a major architectural dislocation at Politiko-Troullia in response to local landscape erosion, possibly due to increased regional precipitation. We present a regional 14C model for Prehistoric Bronze Age Cyprus combining the chronology for Politiko-Troullia with modeled 14C ages from Sotira Kaminoudhia and Marki Alonia, which is bolstered by individual ages from five other settlements on Cyprus. Through the Prehistoric Bronze Age, agrarian villages on Cyprus developed the foundations for the emergence of urbanized settlement and society during the ensuing Protohistoric Bronze Age. Politiko-Troullia, in conjunction with other key settlements on Cyprus, provides a significant independent contribution to increasingly robust Bronze Age 14C chronologies in the Eastern Mediterranean.

In the wake of the demise of the WTO's Appellate Body, there has been a growing trend of states resorting to the dispute settlement mechanisms under their regional trade agreements (RTAs) to resolve international trade disputes. While the vast majority of these mechanisms have never been used, many contain defective procedural provisions that are likely to slow down or completely derail the dispute settlement process should those provisions be invoked. This is particularly true of mechanisms that effectively permit a respondent to block or delay the composition of a panel to hear a dispute. This article examines the issues of ‘panel blocking’ and panel composition delay tactics in RTA dispute settlement with reference to both past and present practice, and provides a textual analysis of a cross-section of existing RTAs to identify procedural defects and prescribe solutions for ensuring timely panel composition in future disputes.

With type 2 diabetes presenting at younger ages, there is a growing need to identify biomarkers of future glucose intolerance. A high (20%) prevalence of glucose intolerance at 18 years was seen in women from the Pune Maternal Nutrition Study (PMNS) birth cohort. We investigated the potential of circulating microRNAs in risk stratification for future pre-diabetes in these women. Here, we provide preliminary longitudinal analyses of circulating microRNAs in normal glucose tolerant (NGT@18y, N = 10) and glucose intolerant (N = 8) women (ADA criteria) at 6, 12 and 17 years of their age using discovery analysis (OpenArray™ platform). Machine-learning workflows involving Lasso with bootstrapping/leave-one-out cross-validation identified microRNAs associated with glucose intolerance at 18 years of age. Several microRNAs, including miR-212-3p, miR-30e-3p and miR-638, stratified glucose-intolerant women from NGT at childhood. Our results suggest that circulating microRNAs, longitudinally assessed over 17 years of life, are dynamic biomarkers associated with and predictive of pre-diabetes at 18 years of age. Validation of these findings in males and remaining participants from the PMNS birth cohort will provide a unique opportunity to study novel epigenetic mechanisms in the life-course progression of glucose intolerance and enhance current clinical risk prediction of pre-diabetes and progression to type 2 diabetes.

OBJECTIVES/GOALS: Our goal is to develop a cost-effective approach for precision medicine treatment by providing computational predictions for new uses of currently available FDA approved, and experimental drugs for NSCLC. METHODS/STUDY POPULATION: Cell Lines: A549 (ATCC- CCL-185) Human epithelial Lung Carcinoma cells, H1792 (ATCC-CRL-5895) Human Lung Carcinoma cells. In Vitro Cytotoxicity Assay: A Vybrant® MTT Cell Proliferation Assay was used. Colony Formation Assay: NCI-H1792, A549 cells were seeded at a density of 500 cells/ dish, then treated with ARS-1620, Osimertinib. The Computational Analysis of Novel Drug Opportunities (CANDO):  Herein, we employed the bioanalytic docking (BANDOCK) protocol within CANDO to calculate the compound-protein interaction scores for a library of 13,218 compounds from DrugBank against a library of 5,317 protein structures from the Protein Data Bank, resulting in a proteomic interaction signature for each compound, and identified Osimertinib as the most likely EGFR/ErbB inhibitor to synergize with ARS-1620. RESULTS/ANTICIPATED RESULTS: ARS-1620 and Osimertinib in combination displays potent anti-tumor activity as evident by a decrease in cell viability with cytotoxicity assays, as well as reduced number of colonies in the colony formation assay for both A549 and H1792 cells. By using CANDO, and cross-referencing the obtained rankings with known experimental information, we have obtained drug predictions within the context of precision medicine. Our preliminary data indicates that EGFR inhibitor Osimertinib may be most structurally similar to KRAS G12C inhibitors overall, compared to other ErbB/ EGFR inhibitors. Validations with human cancer cell lines A549 and H1792 have confirmed that Osimertinib in combination with KRAS G12C inhibitor ARS-1620 may exhibit a synergistic effect in decreasing cellular proliferation and colony formation. DISCUSSION/SIGNIFICANCE: This suggests that this innovative drug combination therapy may help improve treatment outcomes for KRAS G12C(H1792) and KRASG12S(A549) mutant cancers. Cell migration and cell invasion studies in response to treatment with Osimertinib and ARS-1620 are currently ongoing.

We present the first Bayesian 14C modeling based on AMS ages from stratified sediments representing continuous occupation across the Early Bronze III/IV interface in the Southern Levant. This new high-precision modeling incorporates 12 calibrated AMS ages from Khirbat Iskandar Area C using OxCal 4.4.4 and the IntCal 20 calibration curve to specify the EB III/IV transition at or slightly before 2500 cal BCE. Our results contribute to the continuing emergence of a high chronology for the Levantine Early Bronze Age, which shifts the end of EB III 200–300 years earlier than the traditional time frame and increases the length of EB IV to about 500 years. Data from Khirbat Iskandar also help direct greater attention to the importance of sedentary communities through EB IV, in contrast to the traditional emphasis on non-sedentary pastoral encampments and cemeteries. Modeling of AMS data from Khirbat Iskandar bolsters the ongoing revision of Early Bronze Age Levantine chronology and its growing interpretive independence from Egyptian history and contributes particularly to re-examination of the EB III/IV nexus in the Southern Levant.

Gestational diabetes mellitus (GDM) is a global public health problem, and in India, it affects about 20% of pregnancies. India, despite being a tropical country with abundant sunshine has a high prevalence (80%) of vitamin D deficiency (VDD) among reproductive-aged women. Global and Indian evidence links VDD with a higher risk of hyperglycaemia in pregnancy and GDM. VDD has also been implicated in gestational hypertension, preterm birth and poorer offspring health. Global scientific consensus acknowledges the need for maternal vitamin D screening and supplementation, but knowledge gaps exist about optimal blood levels (50–100 nmol/l), and the required vitamin D dosage (400–4000 IU). Diet can provide <10% of the vitamin D requirements, food fortification can deliver limited amounts, and hence optimal antenatal supplementation is key. Prenatal calcium supplements containing 400 IU of vitamin D may be sufficient for calcium absorption and bone health, but may not provide immunomodulatory benefits, including GDM prevention. Increasing evidence calls for higher maternal vitamin D requirements (2000–4000 IU) for skeletal, metabolic and immune health benefits. Current screening and supplementation for maternal VDD in India is low. We need to invest in future studies to determine optimal maternal vitamin D requirements and formulate policies for vitamin D supplementation to prevent GDM. Improving the maternal vitamin D status is an important nutritional priority for policymakers to reduce the large economic burden of non-communicable diseases (10% of India's gross domestic product), and eventually achieve the 2030 UN sustainable development goals.

Experience gained from responding to major outbreaks may have influenced the early coronavirus disease-2019 (COVID-19) pandemic response in several countries across Africa. We retrospectively assessed whether Guinea, Liberia and Sierra Leone, the three West African countries at the epicentre of the 2014–2016 Ebola virus disease outbreak, leveraged the lessons learned in responding to COVID-19 following the World Health Organization's (WHO) declaration of a public health emergency of international concern (PHEIC). We found relatively lower incidence rates across the three countries compared to many parts of the globe. Time to case reporting and laboratory confirmation also varied, with Guinea and Liberia reporting significant delays compared to Sierra Leone. Most of the selected readiness measures were instituted before confirmation of the first case and response measures were initiated rapidly after the outbreak confirmation. We conclude that the rapid readiness and response measures instituted by the three countries can be attributed to their lessons learned from the devastating Ebola outbreak, although persistent health systems weaknesses and the unique nature of COVID-19 continue to challenge control efforts.

The COVID-19 pandemic is giving way to increases in military engagements in health-related activities at the domestic level. This article situates these engagements amid issues of continuity, change, and resistance in contemporary redefinitions of military health roles. It positions the COVID-19 pandemic as a pivotal moment in global health military practice. I identify three emerging trends within national military responses to COVID-19: (1) Minimal technical military support; (2) Blended civil-military responses; and (3) Military-led responses. The dynamics that underpin each type of military involvement follow context-specific military political legacies. These levels of involvement also relate to national public health approaches and the degree of capacity within health care systems. Each identified trend points towards specific trajectories for the future co-constitution of global and local civil-military engagements.

Expansion of cultivated lands and field management impacts greenhouse gas (GHG) emissions from agriculture soils. Soils naturally cycle GHGs and can be sources or sinks depending on physical and chemical properties affected by cultivation and management status. We looked at how cultivation history influences GHG emissions from subtropical soils. We measured CO2, N2O, and CH4 fluxes, and soil properties from newly converted and continuously cultivated lands during the summer rainy season in calcareous soils from south Florida. Newly converted soils had more soil organic matter (OM), more moisture, higher porosity, and lower bulk density, leading to more GHG emissions compared to historically cultivated soils. Although more nutrients make newly converted lands more desirable for cultivation, conversion of new areas for agriculture was shown to release more GHGs than cultivated lands. Our data suggest that GHG emissions from agricultural soils may decrease over time with continued cultivation.

We present two new Bayesian 14C models using IntCal20 that incorporate 17 new calibrated AMS ages for Early Bronze IV Tell Abu en-Ni‘aj and Middle Bronze Age Tell el-Hayyat, located in the northern Jordan Valley, Jordan. These freshly augmented suites of carbonized seed dates now include 25 AMS dates from Tell Abu en-Ni‘aj and 31 AMS dates from Tell el-Hayyat. The modeled founding date for Tell Abu en-Ni‘aj strengthens an emerging high chronology for Early Bronze IV starting by 2500 cal BC, while the end of its habitation by 2200 cal BC may exemplify a regional pattern of increasingly pervasive abandonment among late Early Bronze IV settlements in the Southern Levant. In turn, our modeled date for the Early Bronze IV/Middle Bronze Age transition at Tell el-Hayyat around 1900 cal BC pushes this interface about a century later than surmised traditionally, and its abandonment in Middle Bronze III marks an unexpectedly early end date before 1600 cal BC. These inferences, which coordinate Bayesian AMS models and typological ceramic sequences for Tell Abu en-Ni‘aj and Tell el-Hayyat, contribute to an ongoing revision of Early and Middle Bronze Age Levantine chronologies and uncoupling of their attendant interpretive links between the Southern Levant and Egypt.