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Intrinsic period stability of the cyanobacterial circadian oscillator across in vitro and in vivo conditions
A defining feature of circadian clocks that enables adaptation to Earth’s rotation is their ability to sustain an approximately 24 h period with precision, regardless of environmental factors such as temperature. This remarkable reliability of circadian timing can be reconstituted in vitro using only three cyanobacterial clock proteins: KaiA, KaiB, and KaiC. In vivo circadian rhythms, however, are governed not only by this Kai protein–based oscillator but also by transcription–translation feedback loops and additional clock components. The contribution of the Kai protein–based circadian oscillator to the overall reliability of the in vivo circadian rhythm remains unclear. In this study, we compared over 20 KaiC period mutants with periods ranging from 15 to 60 h under in vitro and in vivo conditions. In both cases, the period was insensitive to environmental conditions, suggesting a compensatory mechanism independent of metabolic state or rhythm amplitude. The ATPase activity of KaiC, the pacemaker of the cyanobacterial circadian clock, exhibited a stronger correlation with in vitro circadian frequency than in vivo circadian frequency. These results indicate that the KaiC ATPase-driven protein-based circadian oscillator inherently encodes a reliable circadian period independent of rhythm amplitude or environmental conditions. This intrinsic property likely plays a critical role in preserving the precision and stability of circadian timing in vivo while being influenced by the intracellular environment.
Abstract WE434: Higher Gestational Weight Gain Is Associated With A Lower Risk Of Incident Diabetes
Objective: To investigate the association between gestational weight gain (GWG) and risk of future diabetes (DM). Methods: This was a retrospective study in an electronic health record-based cohort of pregnancies at a single institution from 1998 to 2016. We included individuals aged ≥18 at the time of their 1st eligible singleton pregnancy and no prior DM diagnosis, with available follow-up data from non-obstetric clinical encounters. DM was defined using diagnostic codes for diabetes (≥ 2 encounters) or HbA1c ≥6.5%. The primary exposure was total GWG between 12 weeks’ gestation and delivery. Secondary exposures included GWG above vs. below the median and 2009 Institute of Medicine (IOM) recommended GWG categories. Follow-up started at delivery of the 1st eligible pregnancy and continued until DM diagnosis, last non-obstetric clinical encounter, date of delivery of an ineligible pregnancy, or 12/31/2024. In unadjusted analyses, we generated Kaplan-Meier curves, stratified by GWG quintile and by median GWG. In adjusted analyses, we fit Cox models with time-varying covariates: gestational age at delivery, BMI at 12 weeks’ gestation, gestational diabetes, nulliparity, marital status, insurance status, race/ethnicity, and zip-code based social deprivation index. Age at delivery of the 1st pregnancy was included as a non-time-varying covariate. Results: We included 37,595 pregnancies in 27,652 individuals (Table 1). The median (IQR) age at delivery was 31.6 years (27.4-35.0). The majority (57.6%) of participants had a BMI <25 kg/m2 at 12 weeks’ gestation. Over a median (IQR) of 11.5 (4.7-17.9) years, 3.2% of individuals (n= 882) developed DM (incidence rate = 2.7/1000 person-years, Figure 1). In the adjusted analyses, each 2 kg increase in total GWG was associated with a lower DM risk (Hazard ratio 0.97 [95% CI 0.942 to 0.997, p= 0.03). GWG ≥ the median (13.2 kg) was associated with lower DM risk (Hazard ratio 0.86 [95% CI 0.745 to 0.999], p= 0.049) vs. GWG below the median. There was no association observed between IOM recommended GWG category and DM risk (Table 2). Conclusions: Higher GWG is associated with a reduced risk of incident DM in the decade after delivery. Further investigation is needed to elucidate the potential mechanisms behind this association.
Abstract WE436: Identifying the Clinical Threshold of C-Reactive Protein-Triglyceride Glucose Index for Risk Stratification of Diabetes
Introduction: C-reactive protein-triglyceride glucose index (CTI) is a novel composite biomarker for type 2 diabetes mellitus. However, its clinical threshold for diabetes across different age and sex groups remains unclear. Methods: Data from 3,517 National Health and Nutrition Examination Survey (NHANES) participants recruited from 2021 to 2023 with complete age, gender, triglyceride (TG), fasting blood glucose (FBG), and high-sensitivity C-reactive protein (hs-CRP) were analyzed. CTI was calculated as 0.421xLn(hs-CRP)+Ln(TG mg/dLxFBG mg/dL/2). Odds ratios (OR) with 95% CI for diabetes across five quintiles of CTI were calculated. A restricted cubic spline (RCS) of CTI with 4 knots was fit for the general population (adjusted for age and sex) and across gender and age groups. The clinical threshold was calculated by the first CTI that had an OR greater than 1. The area under the receiver operating curve (AUC) was calculated for models across all groups. Results: Across 3517 participants, 597 had diabetes. Mean CTI values are higher among diabetic participants compared to those who are not (mean CTI 9.56 vs. 8.62, P<0.001). OR for diabetes from lowest(1) to highest quintile(5) was 1.00 (ref), 1.87(95% CI 1.11–3.26), 2.56 (95% CI 1.56–4.39), 4.79 (95% CI 2.98–8.08), 17.3 (95% CI 11.0–28.8). RCS analysis in the general population and across sex and age groups revealed a dose-response relationship that differed by age group (P = 0.00019) but not by gender (P = 0.103). The CTI threshold where CTI reaches an OR > 1 was 8.75 (all), 8.75 (male), 9.21 (female), 9.95 (<20 years), 9.76 (20-34 years), 9.36 (35-44 years), 8.94 (45-54 years), 8.37 (55-64 years), and 7.84 (65+ years). The AUC for CTI was 0.845 (general population), 0.838 (males), 0.845 (females), 0.977 (<20 years), 0.935 (20-34 years), 0.868 (35-44 years), 0.816 (45-54 years), 0.746 (55-64 years), 0.721 (65+ years). Discussion: Diabetes risk is highest with increasing CTI quintiles. CTI exhibits a dose-response relationship to diabetes, with thresholds varying across age groups but not gender groups. The predictive value of CTI is highest in younger individuals, supporting its potential use for risk-stratifying individuals at an earlier stage of life. Conclusion: CTI is a novel composite biomarker in diabetes with varying clinical thresholds for different age groups. CTI with different thresholds may be used to risk-stratify participants at various age groups through further trials that validate these thresholds.
Abstract TU185: Life’s Essential 8 Protection Against Heart Failure: Metabolomic Insights from the REGARDS Study
Background: Heart failure (HF) is a leading cause of morbidity and mortality. Modifiable health behaviors and factors influence HF development, but pathways linking cardiovascular health to HF remain incompletely understood. The American Heart Association’s Life’s Essential 8 (LE8) score captures cardiovascular health across eight components: diet, physical activity, nicotine exposure, sleep, body mass index, blood lipids, blood glucose, and blood pressure, integrating both behavioral and clinical factors. Hypothesis: We hypothesize that higher LE8 scores are associated with lower HF risk through metabolite-mediated pathways that differ by race and sex. Methods: Complete LE8 scores (scaled 0-100) were calculated at baseline for 12,775 (30% Black; 56% female) Reasons for Geographic and Racial Differences in Stroke (REGARDS) study participants. Incident HF events (n=746) were ascertained over a median 14.8 years. Cox models adjusted for age, sex, race, income, and education estimated LE8-HF associations. Metabolomics quantified 358 metabolites in 2,435 participants. Causal mediation analyses (using bootstrap confidence intervals) evaluated the extent to which metabolites explained the association of LE8 (composite or components) with incident HF. Analyses were further stratified by race and sex. We report metabolites with plausible effect estimates and significant natural indirect effects (p<0.05). Results: Each 10-unit higher overall LE8 score was associated with a 26% lower risk of HF (hazard ratio=0.74, 95% CI: 0.70-0.78). This effect was consistent across sex and race, with hazard ratios ranging from 0.63 to 0.76. Cotinine, a nicotine exposure marker, mediated ~20% of the overall LE8-HF association, and 41% and 33% of the LE8-HF association in men and White participants, respectively. Lysophosphatidylethanolamine (LPE 18:2) isomers mediated 13%-25% of the LE8-HF association. Among LE8 components, dimethylguanidino valeric acid mediated 14% of the blood glucose-HF association in Black participants. Conclusions: Higher LE8 scores consistently predict lower HF risk across multiple metabolic pathways, regardless of sex or race. Nicotine exposure-, glucose-, and lipid-related metabolites partially explain the protective association between cardiovascular health, as measured by LE8, and HF. These circulating metabolites may serve as intermediate biomarkers linking lifestyle and clinical factors to HF, reinforcing LE8 as a comprehensive target for prevention.
Targeted NMR signal enhancement of RNA by site-directed bis-nitroxide labeling
MicroRNAs regulate gene expression through sequence-specific interactions with target messenger RNAs (mRNAs), and their misregulation is a hallmark of cancer. MicroRNA-34a (miR-34a), a key modulator of the tumor suppressor p53, binds the mRNA encoding sirtuin 1 (mSirt1) and adopts multiple conformational states that influence repression efficiency. While such dynamics have been characterized in vitro, extending these studies to cellular environments is hampered by weak signals and substantial background inherent to nucleic acid NMR. To overcome this limitation, we developed a site-directed spin labeling strategy for RNA that enables targeted dynamic nuclear polarization (DNP) signal enhancement. Using the bisnitroxide polarizing agent AsymPol-NCS-SDSL, we conjugated spin labels to specific positions of mSirt1 RNA and annealed them to 13 C, 15 N-cytidine-labeled miR-34a. At 9.4 T, we observed up to 27-fold signal enhancements. The selectivity of polarization transfer within the RNA duplex relative to the surrounding environment could be tuned by matrix deuteration, while doping with paramagnetic metal ions accelerated polarization build-up times, with Cu II proving more efficient than Gd III . This work establishes bisnitroxide-based SDSL as a powerful approach for targeted DNP of nucleic acids, enabling high-sensitivity studies of nucleic acids at concentrations ≤40 µ m and paves the way for structural investigations of microRNA–mRNA interactions in cells.
Abstract TU236: Cumulative Cardiometabolic Risk Increases with A1C Even at Subtherapeutic A1C Levels (5.0% to 5.7%)
Background: Current guidelines link elevated hemoglobin A1C above 5.7% with increased cardiometabolic risk; lipid profile, hyperglycemia, and vascular inflammation all serve as key risk indicators. We aimed to assess whether these indicators correlationally increase with A1C levels within the normal range, affecting clinical decisions made in preventative care. Hypothesis: In a real-world rural cardiology clinic population, do biomarkers of lipid burden, vascular inflammation, and discordance rise significantly within the A1C range 5.0%–5.7%, suggesting early cardiometabolic risk? Methods: We extracted laboratory results for A1C and corresponding non-HDL, LDL, and HDL cholesterol, triglycerides (TG), apolipoprotein B (ApoB) , myeloperoxidase (MPO), lipoprotein(a) (Lp(a)), high-sensitivity C-reactive protein (hs-CRP), homocysteine (Hcy) and insulin resistance score (IRS) levels measured over a 5-year period from 2019 to 2024. Results: Key biomarkers for 2,373 patients with A1C values between 4.0% and 7.0%, and specifically 1,422 patients with A1C 5.0–5.7%, were analyzed. Lipid abnormalities: As A1C rose through 7%, TG values and the percentage of patients with TG >150 mg/dL increased significantly. ApoB-LDL-C discordance rose linearly with A1C from 4.5%–7%, with a 5–10% increase beginning at A1C 5.0%. Vascular inflammation patterns: Both MPO and Lp(a) levels peaked at A1C 6.6% and 6.9%. The frequency of elevated hs-CRP (defined as >2 mg/L) levels, showed a notable positive correlation with A1C from 5.0%–7.0% (R 2 =0.17). In contrast, MPO (R 2 =0.007) and Hcy (R 2 =0.013) had weak correlations with A1C in this range. IRS demonstrated the strongest positive correlation with rising A1C values from 5.0%–7.0% (R 2 =0.339) although not as strong when limited to subtherapeutic levels as hs-CRP. In summary: 33.76% of patients with an A1C of 5.0%–7.0% had one elevated marker, 20.15% had two, 8.49% had three, and a small subset had ≥4. The proportion with two or more elevated markers increased across A1C 5.0–5.7%, reflecting cumulative cardiometabolic burden. Elevated TG, apoB-LDL-C discordance, and hs-CRP were the most common abnormal biomarkers in this range (R 2 =0.70, 0.35, 0.54). Discussion: Despite widespread preventative measures taken by our patient population, trends of increased cardiovascular risk exist at traditionally subtherapeutic A1C values. We conclude that these trends emphasize the importance of proactive medical therapies to overall cardiovascular health.
Abstract TH880: Feasibility and Acceptability of Fitness Testing at the Minnesota State Fair
Introduction: Wearable technology paired with mobile device applications can provide a convenient, low-barrier method to investigate individual fitness and evaluate cardiovascular (CV) health in a precise and equitable manner. In this study, we aimed to assess the feasibility of the University of Minnesota 3-Minute Heart Health app, analyzing technical function and usability feedback. Methods: This study was conducted at the Minnesota State Fair, enrolling healthy, asymptomatic adults. Participants were fitted with a Bluetooth continuous heart rate monitor chest band connected to the 3-Minute Heart Health app. They then performed 2 minutes of vigorous exercise followed by 1 minute of recovery (Figure). Subsequently, they completed a survey including demographic questions and the System Usability Scale (SUS). The SUS is a ten-item survey assessing subjective usability. It is scored using a Likert scale from 0 to 100, with values higher than the industry mean SUS score of 68 serving as a good indicator of user satisfaction. Results: Of the 263 individuals who enrolled, 201 individuals were able to achieve a heart rate necessary to assess CV fitness, a completion rate of over 75%. Twenty-one technical failures were documented, involving an inability to detect heart rate related to issues with Bluetooth connection, band fit, or another unknown problem. A total of 41 individuals (15.6%) were unable to reach an adequate exercise heart rate necessary for the app to provide a heart fitness grade. Of the 201 individuals who completed exercise testing, age ranged from 18 to 90 years. A total of 230 participants completed the SUS survey. Scores indicated that the app was well-accepted; mean SUS score for all survey participants was 75.2. Scores were high across age and education subgroups, including among individuals over 65 years of age (SUS score of 74.1). Participants provided generally positive written feedback after using the app, finding it to be user-friendly. Suggestions for areas of improvement included adding more options for heart rate monitoring, improving the fit of the chest band, and optimizing Bluetooth connectivity. Conclusions: The participant mean SUS score of 75.2 is well above the accepted benchmark of 68, indicating satisfaction with app usability. Going forward, this app may be an accessible screening tool for asymptomatic individuals to evaluate CV fitness without needing to pursue costly, time-intensive procedures and specialty care.
Abstract TU111: Advanced Peripheral Neuropathy is Associated with Death in Older Adults
Objectives: Peripheral neuropathy (PN) is common in older adults, even in the absence of diabetes. The association of PN with mortality in older adults is poorly characterized. We aimed to assess the association of advanced PN with mortality in community-dwelling adults aged 70-95 years. Methods: We conducted a prospective cohort analysis of participants in the Atherosclerosis Risk in Communities (ARIC) Study who underwent the 5.07 Semmes–Weinstein nylon monofilament insensitivity test for PN between 2016 and 2017 (ARIC Visit 6). Advanced PN was defined as lack of sensation at one or more foot sites during the monofilament test. We used Kaplan Meier survival analyses to estimate crude differences in all-cause mortality by PN status, and Cox regression models adjusting for demographics and clinical factors to quantify the association of advanced PN with all-cause mortality, overall and stratified by diabetes status. Results: Among 3,501 participants (median age 79 years; 41% male; 21% Black; 35% with diabetes, median follow up time 4.0 years), 34% had advanced PN. Estimated 5-year mortality was 25.5% (95%CI 23.0%–28.0%) in individuals with advanced PN and 14.2% (95%CI: 12.7%–15.6%) in those without PN (P<0.001). Participants with advanced PN had higher all-cause mortality compared to participants without PN regardless of diabetes status (Log-rank P<0.001; Figure ). After risk adjustment, advanced PN was associated with a higher risk of all-cause mortality overall (HR 1.33, 95%CI 1.15–1.55) and among participants without diabetes (HR 1.53, 95%CI 1.26–1.87). However, the association of PN with mortality was no longer significant among participants with diabetes (HR 1.10, 95%CI 0.87–1.38; P-value for interaction = 0.01). Conclusions: Older adults with advanced PN detected by monofilament testing have a higher risk of all-cause mortality compared to older adults without advanced PN, particularly among patients without diabetes. Reduced foot sensation may be an important screening tool for mortality risk in older individuals regardless of their diabetes status.
Imported, not invented, genes prevail among <i> <i>Escherichia coli</i> </i> ORFans
Bacterial genomes contain numerous ORFans—genes lacking homologs outside the species in which they are found. The source of these genes remains enigmatic because the major mechanism by which new genes originate—by duplication and divergence—is rare in bacteria. The proposed explanations for the birth of ORFan genes include horizontal transfer from sources unrepresented in the databases and rapid divergence from preexisting sequences; however, the lack of direct homology-based evidence has left this issue unresolved. We curated a high-confident set of Escherichia coli -specific ORFans whose distributions were then charted across the species’ pangenome. Based on their patterns of occurrence, ORFan genes could be assigned to one of two modes of origin. The majority were recently acquired via horizontal transfer, with phage transduction making a significant contribution. A smaller fraction of genes emerged via sequence divergence from resident coding genes or de novo from noncoding sequences. Those acquired horizontally are chiefly of unknown function, whereas those arising from resident sequences are primarily involved in defense and membrane-associated activities. This phylogeny-informed approach demystifies the origins of ORFan genes and offers a route toward establishing their source across bacterial taxa.
Abstract TU183: Genome-wide association study identified novel genes involved in transporting and metabolizing blood per- and polyfluroalkyl substances
Introduction: Per- and polyfluroalkyl substances (PFAS) are persistent environmental pollutants known as “forever chemicals” that bioaccumulate and cause adverse health effects. The US Environmental Protection Agency (EPA) has proposed to designate PFAS as hazardous substances and implement measures to eliminate perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS) from food and water sources. Understanding the genetic mechanisms regulating PFAS metabolism and clearance may facilitate the development of novel interventions to reduce their body burden. We aim to identify genetic determinants of plasma PFAS levels. Methods: We performed genome-wide association analyses (GWAS) of plasma PFOA and PFOS among 9992 White participants from the Canadian Longitudinal Study of Aging (CLSA) cohort. After genomic control, genetic variants reaching suggestive significance (P<1E-4) were further evaluated for replication using summary statistics from 6,136 Finland men from the Metabolic Syndrome in Men (METSIM) study. PFOS and PFOA were quantified using the untargeted Metabolon platform in both cohorts. GWAS results from the two cohorts were meta-analyzed using METAL software. Variants achieving nominal significance (P<0.05) in replication analyses and genome-wide significance (P<5E-8) in the meta-analysis and having consistent effect directions were considered significant. Results: As shown in Table , we identified three novel loci associated with plasma PFOS concentrations: a missense (stop gained) variant in ABCG2 (rs2231142), an intronic variant in CYP2C9 (rs112704334), and an intergenic variant (rs148375281). The minor T allele of ABCG2 rs2231142 was associated with higher PFOS levels, whereas the minor alleles of rs112704334 and rs148375281 were associated with lower PFOS levels. No genome-wide significant variants were identified for PFOA. The ABCG2 locus contains genes involved in xenobiotics transportation in the kidney. The CYP2C9 locus contains genes encoding enzymes of the cytochrome P450 family, which are important for xenobiotics metabolism. Conclusion: We identified three novel and functionally relevant loci influencing blood PFOS levels. These findings provide new insight into the genetic regulation of PFAS metabolism and may inform future strategies to reduce PFAS accumulation in humans.
Abstract 30: Gestational weight gain, post-partum weight retention, and women’s mid-life cardiovascular health in Project Viva
Background: Inadequate or excessive gestational weight gain (GWG) is associated with adverse pregnancy outcomes. Less is known about how GWG and post-partum weight retention impact women’s long-term cardiovascular health (CVH). Methods: Participants are from Project Viva, a prospective cohort of pregnant women and their offspring. We defined GWG as the last clinically measured pre-delivery weight from clinical records minus pre-pregnancy weight (from self-report). We defined post-partum weight retention as post-partum weight (measured at 6 months; self-reported at 1 and 2 years) minus pre-pregnancy weight. At the Mid-life Visit (2017-2021), we measured blood pressure, body mass index (BMI), glucose, hemoglobin A1c, cholesterol, and participants self-reported health behaviors (sleep, diet, physical activity, smoking). We derived American Heart Association Life’s Essential 8 (LE8) scores (0-100, higher=better) and used linear regression to examine associations of overall and trimester-specific GWG and post-partum weight retention at 6 months, 1, and 2 years post-partum with overall LE8, its biomedical and behavioral domains, and individual metrics. Models were adjusted for pre-pregnancy body mass index, age of mother at enrollment, and mother’s educational attainment at enrollment. Results: Among 819 women (mean [SD] age 32.6 [5.0] years at enrollment and 51.0 [5.1] years at mid-life; 68% Non-Hispanic White, 15% Non-Hispanic Black), pre-pregnancy BMI was 24.6 (5.1) kg/m 2 ( Table 1 ). Higher GWG (per 1 kg) was associated with a worse BMI score at mid-life for pregnancy overall (β = -0.71, 95% CI: -1.02, -0.39), and for the 1 st (β = -1.07, 95% CI: -1.67, -0.48), 2 nd (β = -1.14, 95% CI: -1.89, -0.40) and 3 rd (β = -0.72, 95% CI: -1.31, -0.13) trimesters individually ( Table 2 ). Higher post-partum weight retention (per 1 kg) at all time points was associated with a worse LE8 biomedical domain (indicating worse CVH), a worse BMI score, and a worse BP score ( Table 3 ). Higher post-partum weight retention at 6 months (β = -0.36, 95% CI: -0.68, -0.04) and 2 years (β = -0.36, 95% CI: -0.67, -0.05) was also associated with a worse blood glucose score. Conclusion: In this prospective cohort, higher GWG was associated with higher mid-life BMI, and higher post-partum weight retention was associated with higher BMI, BP, and worse overall CVH. Minimizing post-partum weight retention may be an important target for optimizing women’s long-term CVH.
Abstract TU136: Quantifying the Rising Burden of Heart Failure Due to Cocaine Use in High-Income Countries: A 34-Year Global Perspective
Background: Cocaine use disorder (CUD) represents a significant, yet often underappreciated, public health challenge in high-income countries (HIC), contributing to substantial cardiovascular morbidity and mortality. Cocaine-associated cardiotoxicity can manifest as Cocaine-Induced Cardiomyopathy (CIC), ultimately leading to Heart Failure (HF). Method: We analyzed prevalence and years lived with disability (YLD) for HF due to CUD, using standardized GBD 2023 tool stratified by age, sex, year and location across HIC from 1990 to 2023. Cocaine exposure was estimated using DisMod-MR 2.1, a Bayesian meta-regression model. The burden attributable to cocaine was derived using the Comparative Risk Assessment framework, integrating exposure prevalence, relative risks, and theoretical minimum risk levels to calculate population-attributable fractions. Results: Between 1990 and 2023, the absolute number of prevalent cases of HF due to CUD increased from 3,028 to 8,608, while YLDs rose from 279.9 to 793.2 in HIC. The age-standardized prevalence rate increased significantly, with an AAPC of 7.07%, and the age-standardized YLD rate grew at 7.08%. Region-wise, the highest AAPC in prevalence was observed in Australasia (8.05%), followed by High-income North America (HINA) (6.71%), Southern Latin America (6.68%), Western Europe (4.39%), and High-income Asia Pacific (3.36%). The peak burden of HF YLDs due to cocaine use was observed in the 30-34 year age group, reaching approximately 0.51 YLDs per 100,000 population while prevalence rate 5.53 per 100,000 in 2021. HINA was the dominant regional contributor, accounting for over 85% of the total YLDs in the young-to-middle-aged adult population in 2021. Sex-wise trends showed that males had a higher AAPC in YLDs (6.17%), while females showed slightly higher AAPC in prevalence (6.17%) vs. males (5.60%), indicating differing trajectories in disability progression and healthcare utilization. Conclusion: HF attributable to CUD has escalated significantly in HIC over the past three decades, with HINA consistently contributing the highest burden. Australasia showed the steepest rise in prevalence, while the peak disability occurred in adults aged 30–34 years. Males experienced greater YLD progression, whereas females showed a faster rise in prevalence.
The conserved fibroblast growth factor receptor–based signaling is required for dendrite regeneration
Both axons and dendrites of a neuron are susceptible to physical insults during stroke and trauma. Unlike axon regeneration, the mechanisms of dendrite regeneration remain largely elusive. In this study, we developed a high-throughput method to induce dendrite injury in the PVD neuron of Caenorhabditis elegans , enabling systematic analysis of regenerative mechanisms. We combined this efficient injury method with RNAi to screen through 825 genes that are highly expressed in PVD neurons. We identified key regulators of dendrite regeneration, including the fibroblast growth factor receptor (FGFR) EGL-15 and its downstream signaling components, regulators of F-actin dynamics, and transcription factors. Moreover, our findings indicate that FGFR/EGL-15 is both necessary and sufficient within the neuron at an early point after injury to initiate branching from the broken dendrite tip, and it is also required in the epidermal tissue to prevent the distal dendrite degeneration. Further analysis revealed that the FGF ligand, EGL-17, functions within the surrounding epithelia for the neuron-specific role of EGL-15. Downstream of EGL-15, the GRB2/SEM-5 adaptor-dependent signaling arm involving GEF/SOS-1, RAS/LET-60, and MAPK-1/MPK-1 is required for dendrite regeneration. In summary, our study identified the conserved FGFR signaling that responds to dendrite injury to initiate dendrite regeneration. Our screening also identified pathways involving F-actin dynamics and transcription factors, which will help the dissection of the dendrite regeneration mechanism in the future.
Abstract WE499: Social Determinants and Accumulation of Cardiometabolic Risk Factors in a Community Cohort: An Inverse Probability of Treatment Weighting Analysis
Introduction: The co-occurrence of cardiometabolic risk factors (CMRFs) is strongly associated with all-cause mortality and incident disability. Although social determinants of health (SDoH), including lifestyle and psychosocial factors, have been recognized as important contributors to chronic diseases, their causal relationships with CMRF accumulation have not been sufficiently investigated. Hypothesis: SDoH are causally associated with CMRF accumulation. Methods: A total of 5,332 participants (2,362 men and 2,970 women, ≥40 years at T0) without cardiovascular disease completed initial (T0; 2012–2015) and six-year (T1) surveys in a Japanese population-based cohort. SDoH variables assessed at both timepoints included physical activity, sedentary time, sleep, alcohol, smoking, weight change, Kessler 6, Lubben Social Network Scale, living arrangement, occupational stress, and education. We analyzed cross-sectional associations between CMRFs scores (sum of hypertension, dyslipidemia, impaired glucose metabolism, and overweight, defined according to clinical guidelines) and SDoH at T1, using inverse probability of treatment weighting (IPTW) based on T0 data to estimate causal effects, applying multinomial logistic regression to estimate odds ratios comparing 1, 2, or ≥3 CMRFs versus 0 CMRFs adjusted for sex, age, and smoking. Results: Weight gain since age 20 showed the strongest association with CMRF accumulation, and recent weight gain within the past year was also positively associated with higher CMRF scores. Past smoking was associated with increased odds of CMRF accumulation, while current smoking showed a paradoxical inverse association, likely reflecting reverse causation whereby individuals with higher disease burden had already quit smoking. The cross-sectional association between current alcohol consumption and CMRFs observed in standard multinomial analysis disappeared after inverse probability of treatment weighting adjustment for T0 confounders. Additional modest associations included low weekday physical activity, high psychological distress, and short sleep duration. Conclusions: Both early adulthood weight gain and recent weight gain are the most robust modifiable risk factors for CMRF accumulation. These findings highlight weight management throughout life, warranting targeted interventions in health screening programs. Further investigation of other social determinants is needed to fully understand their contributions to CMRF development.
Abstract TU263: Association of Difficulty Falling Asleep and Sleep Duration with the Risk of Cardiovascular Events in the General Japanese Population: NIPPON DATA 2010
Introduction: Sleep quality could be an important risk factor for cardiovascular disease, beyond the influence of sleep duration. In particular, difficulty falling asleep, a symptom of poor sleep quality, may be associated with cardiovascular disease; however, evidence in Asian populations is limited. Objective: To examine the association between difficulty falling asleep and incident major cardiovascular events (MACE) beyond the effect of sleep duration in the general population. Methods: We followed 2,507 participants (1,038 men and 1,469 women) enrolled in the NIPPON DATA 2010 study in 2010, excluding those with prior cardiovascular events, until December 31, 2019. Sleep information was obtained from self-administered questionnaires and assessed by difficulty falling asleep (frequent, occasional/rare, none) and sleep duration (short: <6, moderate: 6–<8, long: ≥8 hours). The outcome was incident MACE (stroke, myocardial infarction, PCI, or heart failure). Hazard ratios (HRs) for difficulty falling asleep and sleep duration were estimated using Cox proportional hazards models adjusted for sex, age, BMI, diabetes, hypertension, dyslipidemia, smoking, alcohol use, K6 score, and physical activity index. Both sleep variables were included in the same model for mutual adjustment. Results: During the 8.3-year observation period, 118 MACE occurred (72 in men, 46 in women). The mean age was 58.1±15.8 years. In the overall analysis, compared with those who frequently had difficulty falling asleep, the HRs (95% CIs) were 0.54 (0.33–0.88) for participants who occasionally or rarely had difficulty and 0.51 (0.26–0.99) for those who never had difficulty. For sleep duration, compared with those with moderate sleep duration, the HR (95% CI) was 0.79 (0.49–1.29) for participants with short sleep duration and 1.48 (0.91–2.41) for those with long sleep duration. In analyses stratified by sex, women with long sleep duration showed a higher HR [HR (95% CI) = 2.70 (1.25-5.83)], while this pattern was not observed in men. Conclusions: This study found that participants with frequent difficulty falling asleep had a higher risk of MACE in both sexes, independent of sleep duration, whereas long sleep duration increased the risk among women. Our findings suggest that evaluating difficulty falling asleep as an indicator of sleep quality is important for cardiovascular risk assessment, as well as sleep duration.
Abstract TH957: Performance of Cardiometabolic Polygenic Scores in a high-altitude Peruvian Population
Background: Polygenic scores (PGS) aggregate the effects of multiple genetic variants to estimate disease risk. Limited ancestral diversity in discovery and validation datasets may reduce performance in underrepresented populations. We evaluated the transferability of associations for existing PGS for cardiometabolic phenotypes in a Peruvian population. Methods: We conducted a cross-sectional analysis of 685 participants from Puno, the high-altitude CRONICAS site with substantial Native South American (Quechua and Aymara) ancestry. We evaluated previously validated PGS from large, multi-ancestry, genome-wide association studies for 10 traits: systolic blood pressure (SBP), diastolic blood pressure (DBP), LDL-C, HDL-C, triglycerides, waist circumference, BMI, fasting glucose, fasting insulin, and type 2 diabetes mellitus (T2DM). We also tested PGS associations with clinically defined categorical variables for lipids, waist circumference, and blood pressure. All PGS were mean-centered and standardized. Models were adjusted for age, sex, and ancestry-informative principal components. Bonferroni correction accounted for testing across phenotypes. We estimated ancestry proportions using ADMIXTURE software. Results: Participants were predominantly of Native South American ancestry (93.2%), with minor European (5.8%), African (0.7%), and East Asian (0.2%) contributions. PGS were significantly associated (P < 0.0050) with several cardiometabolic traits after adjustment: SBP increased by 3.80 mmHg (95% CI 2.72–4.88), DBP by 2.78 mmHg (2.11–3.46), LDL-C by 9.38 mg/dL (6.68–12.07), HDL-C by 3.34 mg/dL (2.53–4.15), triglycerides by 13.07 mg/dL (6.72–19.42), and waist circumference by 1.94 cm (1.09–2.78) per 1-SD increase in their respective PGS. PGS for BMI, fasting glucose, fasting insulin, and T2D were not significantly associated with their respective phenotypes. PGS for SBP, LDL-C, HDL-C, and triglycerides were also associated with their respective clinically defined categorical variables, according to expected gradients. Conclusion: In a Peruvian cohort with predominantly Native South American ancestry, PGS derived from multi-ancestry datasets were strongly associated with multiple cardiometabolic phenotypes. These findings underscore the transferability of PGS based on cross-population genetic studies and the need for broader South American representation in genetic studies to further improve PGS performance in these distinct populations.
C4b-binding protein as an antigenic target in the pathogenesis of antiphosphatidylethanolamine autoantibodies
Autoantibodies against phosphatidylethanolamine (PE), a major phospholipid in cell membranes, are associated with symptoms of thrombosis and obstetric complications. A growing body of evidence indicates the involvement of a cofactor for the reactivity of anti-PE (aPE) antibodies. The goals of this study were to identify the putative cofactor and investigate the pathogenic roles of aPE antibodies. Existing ELISA-based assays for detecting aPE antibodies showed that the bovine plasma is a known source of cofactor for the manifestation of reactivity by aPE antibodies. We used affinity pull-down and serial fractionation to purify the cofactor from bovine plasma and identified it as the C4b-binding protein (C4BP), which is a conserved inhibitor in the complement cascade. The human C4BP was subsequently shown to bind PE membrane with specificity and is targeted by aPE antibodies which led to elevated complement activation. Similarly, murine aPE antibodies bind C4BP, hindering its inhibitory function, causing heightened complement activation in vitro and in vivo in a mouse model of renal ischemia and reperfusion, which was antagonized by the coadministration of an anti-C5 antibody. Collectively, the present data identified C4BP with PE-binding specificity and as an antigenic target for aPE antibodies, which cause complement activation. These findings shed light on the pathogenic mechanism of aPE antibodies with implications in diagnosis and therapeutic treatment.
Abstract WE513: Diet Quality and Stage I Hypertension under Contemporary Guidelines: National Trends and Dietary Simulation of Treatment Reclassification
Background: The 2025 AHA/ACC hypertension guideline newly anchors management to PREVENT 10-year risk and calls for a 3-6 month period of nonpharmacologic therapy followed by reassessment before initiating drugs in low-risk adults with stage I hypertension. Nationally representative evidence on diet quality in updated guideline-defined classes is limited. Here we quantify DASH diet adherence across new treatment eligibility classes and run pragmatic simulations to estimate reclassification for stage I at follow-up. Methods: We analyzed data from adults 20-79 years old from NHANES 2013-March 2020, categorized using 2025 AHA/ACC blood pressure cutoffs (Normal, Elevated, Stage I and II). Stage I pharmacologic eligibility followed updated guidelines - immediate pharmacotherapy with clinical CVD, diabetes, or CKD, or with PREVENT 10-year risk ≥7.5%. Diet quality was measured using the DASH diet score from 24-hour recall. For simulation analyses, we identified untreated Stage I adults who were ineligible for immediate pharmacotherapy and DASH non-adherent. We simulated a deterministic diet-only SBP reduction scaled by attenuation factors to account for real-world adherence, holding other risk factors constant. We then recomputed BP category to quantify the proportion and nationally weighted counts spared pharmacotherapy at 3-6 months. All analyses incorporated survey-weighting for national estimates. Results: We analyzed 16,040 adults (weighted N = 217 million) with mean age 46.9 years and 49.3% male. Overall DASH adherence was 8.8% (19.0 million). Across blood pressure categories, DASH adherence declined from normal to higher stages (Figure). Within stage 1, DASH adherence was low across eligibility classes with least adherence noted among those newly eligible for pharmacotherapy via PREVENT risk ≥7.5%. In simulation analyses, a diet-only SBP reduction of ~5 mmHg scaled by an attenuation factor of 0.5, downgraded 12.0% (2.4 million) from ‘Stage I’ to ‘Elevated BP’, sparing pharmacotherapy at follow up. About 88.0% (17.5 million) of stage I adults remained in stage I despite simulated SBP decline, becoming eligible for pharmacotherapy at 3-6 months per updated guidelines. Conclusion: DASH adherence was uncommon and lowest in higher BP categories, underscoring a substantial implementation gap. Simulated improvements reclassified a modest subset of stage I cases, suggesting scalable dietary support could reduce near-term pharmacotherapy initiation.
Abstract TU253: Predictors of Language Strength in State Legislation Related to School Nutrition
Introduction: Dietary habits are a major contribution to the risk of developing chronic disease. School-based nutrition programs play a vital role in supporting children’s nutrition, and state-level policy can shape how these programs are implemented. The specific wording used in legislation can influence how policies are interpreted/enforced. Literature suggests that the use of strong language (e.g. must, shall, required) in child nutrition bills is associated with improved dietary quality. No studies to date have examined predictors of strong language in state-level child nutrition bills; this study addresses that gap by identifying factors associated with its use. Hypothesis: We hypothesized that there would be characteristics and content areas associated with the likelihood of including strong language. Methods: A legislative database (Legiscan) was used to construct a state-level, child nutrition legislation database from 2010 and 2020. Bills were coded for general characteristics (status, year, type, branch introduced, state), content areas (e.g. meal participation, stigma, unpaid meals), and additional factors (funding, enforcement, strength of bill language). Multivariable models were used to identify predictors of strong bill language. Results: Of the 582 bills introduced, 80.1% (n=466) used strong language. In a multivariable model, open/closed campus policies, school garden, unpaid meals, funding, and enforcement were all significantly related to the use of strong language. Bills addressing open/closed campus policies had 9.5 times higher odds of using strong language (CI:1.12, 80.21) than those that did not. Bills addressing school gardens were associated with 4.4 times higher odds of strong language (CI:1.78, 10.69) compared with bills that did not. Bills addressing unpaid meals had significantly lower odds of using strong language (OR = 0.16, CI:0.05,0.54) compared with bills that did not address unpaid meals. Bills including funding provisions had 60% lower odds of using strong language than those without funding (CI:0.24, 0.96). Bills including enforcement provisions had significantly lower odds of using strong language (OR = 0.27, CI:0.12, 0.64) compared with bills without enforcement provisions. Conclusion: Both supportive and limiting predictors of strong language were identified. Understanding how bill topics are associated with bill language strength can inform the drafting of child nutrition policy and guide future advocacy efforts.
Abstract TH954: High, even more than low, ankle-brachial index (ABI) associated with elevated risk of incident cardiovascular disease and all-cause mortality: The Hispanic Community Health Study/Study of Latinos (HCHS/SOL).
Background: Low and high ankle-brachial index (ABI) values are associated with increased cardiovascular disease (CVD) risk, yet their prognostic value in Hispanics/Latinos has not been specifically studied, despite having unique vascular risk factor profiles compared to other races/ethnicities. Aim: We aimed to assess the prospective association of ABI with incident CVD and all-cause mortality in Hispanics/Latinos. Methods: We analyzed data from 8,000 adults (mean age 56, years; range, 45–74 years, 56% female) from the Hispanic Community Health Study/Study of Latinos (HCHS/SOL). The ABI was calculated from relevant extremity systolic blood pressures obtained at baseline (2008–2011) and modeled both continuously (cubic splines) and categorically (≤0.90, 0.91–1.00, 1.01–1.10 1.11–1.20 [reference], 1.21–1.30, and >1.30). Incident fatal and nonfatal CVD events (myocardial infarction, stroke, or heart failure) and all-cause mortality were assessed through 2021. Hazard ratios (HRs) were estimated using Cox models adjusting for confounders. We also conducted subgroup analysis by age, sex, smoking, diabetes, systolic blood pressure and kidney function. Results: Over a median follow-up of 11.6 years, there were 312 CVD events and 601 deaths. The average ABI was 1.08. High ABI (>1.30) vs. ABI 1.11–1.20 was significantly associated with an increased risk of incident CVD and all-cause mortality independently of potential confounders (HR 3.00, [95%CI: 1.40–6.44] and 2.70 [1.57–4.63], respectively). In contrast, low ABI (≤0.9) was significantly associated with all-cause mortality (HR 1.62 [1.04–2.54] but not with incident CVD. The results were largely similar when ABI was modeled continuously (Figure) with lowest risk at approximately ABI 1.10. The results were generally consistent across subgroups. Conclusion: High ABI was more strongly associated with mortality and incident CVD than low ABI in this Hispanic/Latino cohort. These findings have implications for interpreting ABI in this underrepresented population.