The Innovation

Particulate matter exposure has a direct impact on airways diseases, such as asthma and chronic obstructive pulmonary disease (COPD), and over the next 30 years, rising particulate air pollution is expected to increasingly affect disease outcomes. We identified transcriptional mechanisms of particulate exposure in the airway epithelium that connect with disease risk using genetics and multiomics. We first defined and compared rapid-transient nascent transcription responses across particulate exposures. Using hyaluronic acid metabolism as a prototype, we showed that rapid-transient responses to particulates were relevant to steady-state mRNA expression and COPD pathobiology. We then found genetic links between nascent transcription responses and asthma or COPD risk by associating single nucleotide polymorphisms (SNPs) with disease in the All of Us study. By combining nominal association statistics, pre- and post-association filters, and rigorous external validation, we identified SNPs associated with disease across multiple ancestries and cohorts. We then derived epigenetic and gene regulatory mechanisms from these SNPs. Our results highlighted plausible transcriptional mechanisms of disease, such as regulation of TOMM7 expression by rs13243243. By applying detailed transcriptional analysis to study particulate exposures, we identified novel SNPs and genes that define gene-environment interactions for airways disease.

Fine particulate air pollution (PM2.5) in the United States has fallen by roughly half since 2000, yet linked health outcomes such as diabetes and childhood ADHD have not improved in parallel. One reconciling possibility is that pollution exposure in early life produces health effects that emerge only years or decades later, after pollution itself has declined. Using two decades of U.S. county-level data, we relate annual PM2.5 estimates to birth outcomes, diabetes prevalence, and small-area estimates of childhood attention-deficit/hyperactivity disorder (ADHD) across short and long time scales. Within counties, changes in low birth weight rates are associated with changes in PM2.5 during the same year and the year prior to birth. At longer time scales, cross-county comparisons show that PM2.5 exposure is associated with higher prevalence of adult diabetes and ADHD after approximately a decade. Together, these patterns suggest that population-level health risks from air pollution may persist over decades, even as pollution itself declines.

Background Wildfire activity and smoke exposure are increasing worldwide because of climate and land-use change. Although fine particulate matter (PM2.5) may impair pulmonary immune defences against tuberculosis (TB), population-level evidence remains limited. We estimated the effect of wildfire-related PM2.5 exposure on TB notification rates in Brazil. Methods We conducted a nationwide panel study linking municipality-level monthly TB notifications from Brazil's SINAN system with wildfire-related PM2.5 estimates from GEOS-Chem simulations across 5,545 municipalities (2003-2023). We estimated the impact of high-exposure days (PM2.5 > 25 g/m3) on monthly TB notifications using Poisson regression with fixed effects for municipalities, state-by-year, and state-by-month, controlling for time-invariant differences, secular trends, and seasonality. Distributed lag effects were estimated over 1-24 months before notification. Models accounted for meteorological conditions, GeneXpert diagnostic coverage, and spatial correlation using Conley standard errors. We computed attributable fractions among exposed municipality-months (AFE). Sensitivity analyses evaluated alternative PM2.5 thresholds (15 and 35 g/m3), co-pollutants, and agricultural expansion. Findings From Jan 1, 2003 to Dec 1, 2023, 1,758,982 TB cases were reported. Of these, 353,319 (20.1%) had at least one high-exposure day (PM2.5 > 25 g/m3) 1-24 months before notification. An additional 14 high-exposure days over the 24-month lag period was associated with an average monthly increase of 2.9% [95% CI: 0.9-4.9%] in TB notification rates. Effects peaked at 13 months (IQR: 11-14) prior to notification. Results showed a dose-response relationship across PM2.5 thresholds and were robust to controlling for NO2, O3, and agricultural expansion. Overall, wildfire-related PM2.5 exposure accounted for 2.1% [0.7-3.5%] of TB notifications in exposed municipality-months, corresponding to 7,802 [2,612-12,544] attributable cases. The AFE reached 10.7% [7.1-14.0%] in Pantanal and 7.3% [6.1-8.5%] in Amazonia, areas most impacted by wildfires. Interpretation Wildfire-related PM2.5 exposure may represent an increasingly important and modifiable risk factor for TB. As wildfire activity increases across many regions of the world, these findings highlight the need for integrating air quality into climate adaptation and TB control strategies.

While exposure to volatile organic compounds such as ethylene dichloride and vinyl chloride monomer is a well-established cause of liver disease, particularly hepatic hemangiosarcoma, characterizing real-world exposure profiles in communities surrounding industrial centers remains challenging. Calvert City, Kentucky (population ~2,500), provides a unique setting characterized by both active industrial emissions and legacy sources of air toxics. To address these complexities, this method paper describes the framework for the Biomonitoring and Environmental Assessment for Community Outreach and Neighborhood Safety (BEACON) study. By utilizing a novel, multi-dimensional exposure assessment strategy, BEACON aims to characterize air toxic exposures and provide actionable data for community health and safety. For the BEACON study, we will leverage Kentucky Department of Air Quality measures of air toxics, analyze urine samples in a small cohort of community volunteers, analyze community urine via wastewater in an adjacent community, geocode citizen odor reporting, assess blood markers in wildlife, survey small and large animal veterinarians in the area for anomalies in morbidity and mortality, and work with the regional health system to enhance vigilance for health issues associated with toxicants present in the area. In addition, blood samples will be collected at three time points and biobanked for future analyses. Efforts will be made to link this study to additional large-scale long-term cohorts where possible. Throughout the project, community engagement will play a critical role by raising awareness, fostering collaboration, and ensuring that the voices of affected residents are heard.

Background: Lead exposure is estimated to cause approximately 3.5 million premature deaths a year, yet the key ongoing sources of lead exposure are unclear. Methods: We estimated the contribution of dietary lead intake to global blood lead levels (BLLs) for 7-year-old children and 22-year-old adults by applying the All-Ages Lead Model (AALM) to calculate blood lead levels (BLLs) based on 25 total diet studies (TDS) that quantify dietary lead intake across 46 countries. Results: For children, the population-weighted average dietary lead intake in low- and middle-income countries (LMICs) (32.0 g/day) was found to be more than three times higher than in high-income countries (HICs) (9.3 g/day), and more than 10 times higher than the FDA reference level for children (2.2 g/day). The average impact on BLLs for children is estimated to be near 29 g/L in LMICs and near 12 g/L in HICs. Averaged across the TDS data, vegetables (27%) and cereals (24%) were found to contribute the most to dietary lead. Conclusions: While there are limitations associated with biokinetic modelling and the TDS data from LMICs, these results suggest that the contribution of dietary lead intake to global lead exposure is in the region of 40 to 50%, suggesting, in turn, that dietary lead intake is likely a major global driver of lead poisoning. Lead absorbed from the environment into food crops is expected to be the key driver of dietary lead. Current regulatory levels for maximum lead concentrations in foods (0.05-0.3 mg/kg) are out-of-date and may imply a dietary lead intake of 200 g/day, far higher than the FDA reference level (2.2 g/day). Collecting representative TDS data in high lead burden countries should be a priority. Further research is also recommended on upstream lead sources and pathways of lead uptake in plants, driving global food contamination.

Background: Surgical care centralization in the U.S. delays access and increases carbon emissions. Global targets suggest patients live within 2-hours of a surgical facility. This study quantifies the environmental impact of travel for cataract surgery in rural Michigan and models the potential emissions reductions from decentralizing surgical and follow-up services. Methods: A retrospective, cross-sectional study analyzed electronic medical records from a rural Michigan ophthalmology practice (March-November 2023). We calculated travel distances using population-weighted centroids and estimated emissions using U.S. Department of Energy vehicle data. A k-means clustering model optimized additional facility placement, and a gradient analysis identified optimal numbers for decentralization points, for emissions reductions. Results: The 920 patients traveled a median of 55.45 km (IQR: 43.33-88.20 km) for surgery and 55.07 km (IQR: 43.54-87.82 km) for follow-up visits, generating Total Surgical Access Emissions (TSAE) of 57,168 kgCO2; (median of 59.20 kgCO2; IQR: 32.31-81.87) under the centralized model. The k-means decentralization model and gradient analysis identified 7 hospitals and 9 clinics, respectively, as the optimal expansion points, reducing emissions by 34.07% (19,475 kgCO2 saved) and 39.52% (22,590 kgCO2; saved). The Surgical Access Carbon Impact (SACI) model demonstrated that achieving two-hour access to clinic services reduced excess emissions by 54.7%. Sensitivity analyses using fuel-efficient vehicles (Toyota Prius and Tesla Model 3) or reducing follow-up visit frequency reduced emissions by 54.03% (30,888 kgCO2) and 25.83% (14,768 kgCO2), respectively. Conclusion: Decentralizing surgical services in rural U.S. settings could cut travel-related emissions by up to 40%, significantly reducing healthcare-related carbon footprints while improving timely access to care. The SACI metric provides a novel framework for integrating environmental sustainability into U.S. health policy and service planning

Background: Randomized controlled trials have shown that computed tomographic (CT) screening reduces lung cancer mortality. Improved identification of at-risk groups, by leveraging non-smoking risk factors, could help refine screening selection. Aim: To evaluate polygenic risk scores (PRSs) and ambient air pollution (AAP) exposure for risk stratification in the NELSON lung cancer screening cohort. Methods: Two PRSs (PRS-McKay/PRS-Byun) and several AAPs (including nitrogen dioxide, ozone, and particulate matter [PM]) were assessed in the NELSON lung cancer screening trial (N=7,364). PRSs were validated in the Rotterdam Study (N=11,493). Associations with lung cancer, mortality, screening results, and discriminative ability to distinguish lung cancer were evaluated. Results: PRS-McKay and PRS-Byun were associated with lung cancer (odds ratio [OR] per SD [95%CI]: 1.22 [1.08-1.37] and 1.28 [1.13-1.44], respectively) and lung cancer-specific mortality (OR [95%CI]: 1.24 [1.05-1.47], for both), but not with non-lung cancer mortality (OR [95%CI]: 1.01 [0.94-1.10] and 1.03 [0.95-1.12], respectively). Exposure to PM2.5 was associated with lung cancer (OR [95%CI]: 1.11 [1.01-1.22]). PM constituents were associated with adenocarcinoma, particularly PM10 (OR [95%CI]: 1.16 [1.01-1.32]) and ultra-fine particles (OR [95%CI]: 1.16 [1.04-1.30]). PRS and AAP added modestly to the discriminative ability for lung cancer on top of pack-years, age, and sex (area under the curve [95%CI]: 0.659 [0.624-0.695] vs. 0.643 [0.608-0.679]). Conclusions: PRSs and exposure to PM were associated with lung cancer in a high-risk screening population. The primary potential of PRSs may reside in refining lung cancer screening selection toward individuals at higher risk of dying from lung cancer specifically.

Here, we present the first graph-based Korean Pangenome Reference (K-PanRef), constructed from 14 healthy Korean individuals. K-PanRef comprises 13 high-quality diploid Korean genome assemblies (mean QV ~62.0) and KOREF1-G-TTAGGA, the first complete Korean reference genome. Integration of these assemblies generated a ~3.2-Gb pangenome graph containing ~39.3 million nodes and ~53.8 million edges, with the accumulation of common sequences (frequency [≥]10%) reaching a plateau. Additionally, K-PanRef contains ~4.3 million Korean-specific small variants and ~76.0 thousand Korean-specific SVs absent from the Chinese and human pangenome references, improving the representation of Korean genetic diversity relative to these references. To evaluate its utility for short-read-based SV analysis, we genotyped 75 whole-genome sequencing (WGS) samples, including 15 patients with early-onset myocardial infarction (MI). Although constructed entirely from healthy genomes, K-PanRef supported the identification of putative disease-relevant SVs in this exploratory application. K-PanRef-based genotyping identified ~95.6 thousand small variants and 820 SVs observed only in the early-onset MI samples. Among the early-onset MI-group SVs, 491 were absent from public databases, suggesting that they may represent previously unrecognized candidate variants related to early-onset MI. Of these, 164 SVs overlapped 134 genes, of which 89 had reported associations with 42 cardiovascular diseases or traits, including eight genes previously linked to MI. Together, these results establish K-PanRef as a valuable resource for representing Korean genetic diversity and enabling more comprehensive discovery of population-specific and novel putative disease-relevant variants from short-read sequencing data.

Environmental exposures are major modifiable determinants of human aging, yet the evidence remains fragmented across organ-agnostic summaries and rarely confronts population inequity. Here we present an exposomic atlas of pan-organ aging in ~300,000 UK Biobank adults, mapping 164 environmental and behavioural exposures onto biological aging of the whole body and nine organ subsystems. Comprising 1,476 systematically tested exposure-subsystem associations, the atlas reveals that environmental effects on human aging are pervasively organ-specific, with 65.9% of exposures acting divergently across organ subsystems. This landscape resolves into nine navigable modules that preserve organ selectivity, predict 23 major age-related diseases, and expose distinct dimensions of health inequity. In-silico analyses further show that priorities for ameliorating aging are target-dependent rather than universal, diverge markedly from the whole-body ranking (Kendall's {tau} = 0.52 to 0.39), reorder substantially across population strata, with findings externally validated in an ethnically distinct cohort. The atlas establishes an organ-resolved and target-aware foundation for precision environmental health.

Exposure to higher levels of ambient air pollution during pregnancy has been linked to multiple adverse pregnancy outcomes. However, studies on acute exposures and reduced gestation length have reported inconsistent findings. This project aims to examine the acute association between ambient air pollution and preterm (28-36 gestational weeks) or early-term (37-38 gestational weeks) births. Daily concentrations of 12 air pollutants, based on bias-corrected numerical model outputs, were linked to vital records of singleton live preterm and early-term births from 2005-2017 in California, Florida, Georgia, Kansas, Nevada, New Jersey, North Carolina (2005-2015), and Oregon. Under a time-stratified case-crossover design, odds ratios (OR) were estimated via conditional logistic regression with adjustment for risks among ongoing pregnancies, meteorology, time trends and federal holidays. We estimated cumulative associations up to a 6-day lag using distributed lag models. Risk estimates per interquartile range (IQR) increase in exposure were pooled across states using inverse-variance weighting. Our study included 1,085,162 preterm and 3,901,185 early-term births. We observed positive associations between 0-2 day cumulate exposure to several air pollutants and early-term births, including NO2 (OR: 1.0023, 95% CI:1.0010, 1.0037 per 7.1 g/m3 increase), PM2.5 (OR=1.0022, 95% CI: 1.006, 1.0038 per 4.6 g/m3 increase), PM2.5 organic carbon (OR= 1.0026, 95% CI: 1.0013, 1.0039 per 1.7 g/m3 increase) and PM2.5 elemental carbon (OR=1.0025, 95% CI: 1.0014, 1.0035 per 0.26 g/m3 increase). Associations with preterm birth were mostly null. In conclusion, we found positive associations between short-term air pollution exposure, including PM and major PM2.5 components, and risks of early-term birth.

ABTRACT Background Children from low- and middle-income countries are particularly vulnerable to air pollution, a major environmental health risk, due to the immaturity of their lungs and their proximity to sources of household pollution. This study aimed to investigated the effect of exposure to biomass combustion through domestic and maternal occupational activities on respiratory health of children living in disadvantaged urban areas of Abidjan, Cote dIvoire. Methods Between February and December 2023, we conducted a cross-sectional observational study among children <16 years from households of women using biomass fuel for cooking (Group (G) 1), engaged in occupational fish smoking activities (G2), or primarily using gas for domestic cooking (G3). We assessed reported respiratory symptoms through standardized questionnaires and the presence of lung function impairments (LFI) though pulmonary function tests (spirometry and Rint). We assessed the association between study groups and key covariates with respiratory symptoms and LFI using mixed-effects regression models. Results Of 210 children enrolled - 119 (56.8%) female, median age 9 (6-12) years, 82 (39.0%) in G1, 47 (22.4%) in G2, and 81 (38.6%) in G3 - 15 (7.1%) reported wheezing in the last 12 months, 82 (39.0%) reported dry cough at night, 9 (4.9%) presented with dyspnea and 5 (2.7%) had chest pain on clinical examination, for an overall proportion of children with reported respiratory symptoms of 43.8% (92/210). Of 176 children who underwent pulmonary function testing, 59 (33.5%) had LFI detected, including 34 (45.9%) in G1, 8 (22.2%) in G2, and 17 (25.8%) in G3 (p = 0.011). Study group was associated with respiratory symptoms (G1 vs G3; aOR 3.82, 95% CI 1.68-8.68; p < 0.001), as well as with LFI (p = 0.042). Girls were at greater risk of LFI than boys (aOR 2.69, 95% CI 1.24-5.80; p = 0.012). Children whose mothers used charcoal or wood as cooking fuel had higher odds of respiratory symptoms (OR 2.61, 95% CI 1.22-5.58; p = 0.013) but no association was found with LFI (p = 0.459) compared with unexposed children. Conclusion Respiratory symptoms and lung function impairments were highly prevalent among children living disadvantaged, especially when mothers cook with wood or charcoal. Targeted maternal awareness and broader interventions to reduce household air pollution in disadvantaged urban areas are urgently needed to protect long-term respiratory health.

Classical swine fever (CSF) is a highly contagious disease caused by Classical swine fever virus (CSFV), posing a serious threat to the global swine industry. This study aimed to investigate the effect of CSFV on differential genes of histone lactylation at the H3K18 site in the PI3K-AKT signaling pathway. The site with the most significant change in histone lactylation antibody level was screened by Western blot. Omics analysis was performed using CUT&Tag technology to identify differential genes in the PI3K-AKT pathway between the CSFV-infected group and the mock group, followed by validation using RT-qPCR. Functional analysis of significantly differential proteins was conducted, and the protein expression level of THBS4 was detected by Western blot. The results showed that after CSFV infection of 3D4/21 cells, the H3K18la site exhibited the most significant difference in antibody level. A total of 8,859 differential genes at the H3K18la site were identified by CUT&Tag analysis, including 6,349 up-regulated genes and 2,510 down-regulated genes. Further focusing on the PI3K-AKT signaling pathway, 10 differential genes were identified, comprising 6 up-regulated genes and 4 down-regulated genes. Compared with the control group, the mRNA expression levels of CD19, LAMA1, PDGFRA, BDNF, ANGPT4, and THBS4 were up-regulated in the CSFV-infected group, while FOXO3 and NRTN were down-regulated. Western blot results showed that the protein expression level of THBS4 increased after CSFV infection. These findings lay an important foundation for understanding the molecular mechanisms regulating viral replication and immune evasion, and have significant scientific implications and potential application value.

Cryogenic-electron microscopy (cryo-EM) has revolutionized the field of protein structural biology. The structures of large membrane proteins are now routinely determined by cryo-EM to near atomic resolution. However, in the medium resolution range of cryo-EM maps (>[~]2 [A]), negatively charged sidechains of acidic residues are not well-resolved due to the negative electrostatic potential of the region. This may lead to incorrect sidechain models for residues like glutamic acid or aspartic acid that are central for proton transfer activity in various respiratory and photosynthetic enzymes. We previously proposed that the acidic residues with weak or non-existent cryo-EM density can be modeled to represent their low proton affinity conformations. Here, we tested this hypothesis on a larger data set of acidic amino acid residues in two high-resolution respiratory complex I structures. By using faster sidechain modeling and proton affinity prediction tools, we created a workflow that generates sidechain conformations of selected amino acid residues. We validated the sidechain conformation predictions by Q-score analysis and atomistic molecular dynamics simulations in different charged states. The proposed workflow provides a way to rapidly obtain sidechain conformations of acidic residues with weak cryo-EM densities and can be integrated into the existing cryo-EM modeling pipelines to speed up sidechain rotamer prediction.

The impact of weather on the health of Americans and the American health system is substantial. Using available health and economic data, we developed a data-driven scenario that describes a compounded heat emergency in an archetypal community in the United States. We then characterize the potential human and economic costs of such a heat emergency to demonstrate the widespread impact on health outcomes, health systems, and society.

Despite the significant technical advancement in spatial transcriptomics, its clinical usage is largely untapped. Here, we develop an integrated system, ENDO-Genome, for minimally invasive in-body transcript sampling to facilitate live spatial transcriptomic analysis of human internal organs. This is achieved by integrating a nanoarrayed biochip with existing endoscope to perform pressure-sensor-calibrated "Touch & Go" RNA extraction directly from human internal organs, including the highly vascularized liver or kidney, without the need for tissue biopsy, voiding any bleeding risks. By a demonstration using gastrointestinal endoscopy, multiplexed landscape of 55 mRNA transcripts was obtained from multiple locations of human intestinal tract via a 5-minute operation in routine examinations. Benefiting from a sequencing-free approach, each assay costs less than 10 US dollars. For the clinical study involving 15 Crohn' s disease (CD) patients, no complication case was reported out of 47 ENDO-Genome operations, showcasing the gentle deposition and excellent safety of the technique. The live spatial transcriptomics provides direct in vivo pictures of the heterogenous spatial transcriptional programs underlying CD pathological response at different intestinal locations, revealing distinct ileal phenotypes. This is manifested by unique microscale scattering of inflammation gene clusters, along with the discovery of a tissue-specific cooperative mechanisms between inflammation and RNA methylation regulations at single- or multi-cell scales.

Short-term environmental exposures have been linked to cognitive and attention-related outcomes, but the robustness of these associations remains uncertain. We linked daily weather and air-pollution exposures to repeated measures of subjective cognitive difficulties and attention-related outcomes among participants in the All of Us Research Program from 2018 to 2024. Associations were evaluated using complementary longitudinal and causal-inference approaches, including fixed-effects, lagged-exposure, and event-study analyses. Machine-learning methods were used to characterize heterogeneity and latent psychosocial structure, and findings were independently evaluated using 2024 Behavioral Risk Factor Surveillance System data. Several environmental exposure measures were associated with cognitive outcomes in pooled analyses; however, most associations attenuated substantially after accounting for within-location temporal variation. In contrast, mental-health burden, loneliness, and impaired social functioning remained consistently associated with subjective cognitive difficulty across analytical approaches. Similar patterns were observed in the validation dataset. These findings suggest that some observed environmental associations may reflect broader geographic and contextual differences rather than short-term environmental effects. Overall, psychosocial factors demonstrated more consistent associations with subjective cognitive difficulties than short-term environmental exposures across multiple analytical frameworks and independent datasets.

Filovirus outbreaks caused by Ebola virus (EBOV) and Marburg virus (MARV), pose severe global health threats characterized by high rates of fatal hemorrhagic fever. While species-specific vaccines and therapeutic monoclonal antibodies are approved for Zaire ebolavirus, broadly-active therapeutics remain unavailable, leaving populations vulnerable to MARV and other pathogenic Ebola species, such as Bundibugyo (BDBV) and Sudan (SUDV) ebolaviruses. Here we report a computationally guided, infectious virus validated screening platform for the rapid discovery of broad-spectrum filovirus antivirals. By leveraging quantitative structure-activity relationship (QSAR) models, we screened 142,382 compounds in silico to prioritize 125 high-potential candidates. Subsequent dose-response and viability profiling identified 23 compounds exhibiting potent, low-micromolar pan-filovirus activity and favorable cytotoxicity profiles. Molecular docking indicates these compounds target conserved structural and functional domains--primarily the VP35 and L proteins--which may disrupt essential viral replication and immune antagonism. Furthermore, systematic combinatorial screening revealed three highly synergistic compound pairs, notably NCGC00113249-01 and NCGC00118008-01, demonstrating robust cross-species efficacy. By targeting conserved vulnerabilities across the filovirus family, this integrated in silico and in vitro pipeline provides a scalable framework to rapidly nominate and optimize synergistic therapeutic regimens against both endemic and emerging viral threats including BDBV. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=74 SRC="FIGDIR/small/737586v1_ufig1.gif" ALT="Figure 1"> View larger version (34K): org.highwire.dtl.DTLVardef@1251baorg.highwire.dtl.DTLVardef@b3a2feorg.highwire.dtl.DTLVardef@191d314org.highwire.dtl.DTLVardef@b8f710_HPS_FORMAT_FIGEXP M_FIG C_FIG

Enterovirus D68 (EV-D68) is a non-polio picornavirus that has caused increasing rates of severe respiratory illness and acute flaccid myelitis in children worldwide this century. There are no approved vaccines or antivirals for EV-D68. Thus, we conducted a crystallographic fragment screening (CFS) and a high-throughput screening (HTS) biochemical assay against the EV-D68 RNA-dependent RNA polymerase 3D (3Dpol) to identify ligandable sites and non-nucleoside compounds that can spearhead anti-enteroviral drug discovery. The CFS, involving 650 fragments, identified 68 hit compounds (~10% hit rate) distributed across 3Dpol, including the functionally relevant sites RNA template channel, Active site, and RNA primer channel, and the previously unknown "Thumb site II" and "Index-middle finger pocket". Inhibition assays confirmed that compounds binding to each site can inhibit EV-D68 3Dpol activity. The HTS, a fluorescence-based PicoGreen biochemical assay, permitted screening 50,000 compounds of the ChemBridge Premium Library (0.77% hit rate). After a second-round dose-response screening, we identified 5-aminoindazole as a promising scaffold that inhibits EV-D68 3Dpol, including hit-to-lead compound 727590, which displayed an IC50 value of 25 M and preliminary structure-activity relationships. These hits offer amenable starting points for discovery and development of non-nucleoside inhibitors and provide opportunities for structure-based drug design against enteroviruses. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=107 SRC="FIGDIR/small/737532v1_ufig1.gif" ALT="Figure 1"> View larger version (35K): org.highwire.dtl.DTLVardef@14a54a6org.highwire.dtl.DTLVardef@fb6621org.highwire.dtl.DTLVardef@ee2e2aorg.highwire.dtl.DTLVardef@118f91d_HPS_FORMAT_FIGEXP M_FIG Created with biorender.com and PyMOL Molecular Graphics System, version 2.5.0. Schrodinger, LLC. C_FIG

Background: Current chemical mixture approaches are largely data-driven without considering shared biological mechanisms among mixture components, highlighting the need for biology-informed approaches. Objectives: We constructed an integrated measure of a chemical mixture's oxidative stress potential and assessed its association with mortality in the US population. Methods: The sample comprised 4,574 adults ([&ge;] 20 years) from National Health and Nutrition Examination Survey (NHANES) 2005-2010. To obtain robust estimates, we performed 1,000 repeated random 50:50 splits into training and testing sets. In each training set, we used survey-weighted quantile g-computation to model serum gamma-glutamyl transferase (GGT), an oxidative stress biomarker, as a function of a 30-chemical mixture (blood metals, urinary polycyclic aromatic hydrocarbons (PAHs), pesticides, phenols/parabens, and phthalates), adjusting for sociodemographic, behavioral, and dietary factors. We then applied the fitted model from each training set to the corresponding testing set to derive the environmental risk score for oxidative stress (ERSOS), defined by predicted GGT values. Associations of ERSOS with all-cause, cardiovascular, and cancer mortality over 11 years of follow-up were estimated in the testing sets using survey-weighted Cox proportional hazards models and summarized across the 1,000 repeated splits. Results: Chemicals with the largest positive weights in quantile g-computation included mono-(2-ethyl-5-hydroxyhexyl) phthalate, mono-2-ethyl-5-carboxypentyl phthalate, 2-hydroxyfluorene, methyl paraben, and benzophenone-3; chemicals with the largest negative weights included mono-(2-ethyl-5-oxohexyl) phthalate and PAH metabolites (1-hydroxynaphthalene, 3-hydroxyphenanthrene, and 3-hydroxyfluorene). The median correlation between observed and predicted GGT in the testing sets was 0.43 (2.5th, 97.5th percentiles: 0.40-0.48). A one standard deviation increase in ERSOS was associated with a median hazard ratio of 1.60 (2.5th, 97.5th percentiles: 1.01-2.57) for cardiovascular mortality. No associations were found for all-cause mortality or cancer mortality. Discussion: The proposed survey-weighted quantile g-computation approach may help estimate biology-informed chemical mixture effects in complex survey data, supporting the potential utility for population-generalizable environmental mixture research.

Chemotherapy treatment of colorectal cancers (CRC) using cisplatin (CDDP) encounters problems of drug resistance by the cancer cells and cytotoxicity to normal cells, highlighting the urgent need for joint therapeutical strategies. Selenium-enriched rapeseed extracts exhibit anti-cancer effects but the bioactive components and mechanisms remain unclear. Here, we applied different solvents to fractionate the extracts from Selenium-enriched rapeseed and found that the water extract (WE) fraction significantly enhanced the cytotoxic effect of CDDP on cancer cells but no damage on normal cells. HPLC-ICP-MS analysis revealed that methylselenocysteine (MSC) and selenocystine (SeCys2) were the main selenium speciation in WE. Through cell biology and integrative multi-omics analysis, we found a synergistic anti-CRC cell effect when combining CDDP with MSC, sulforaphane (SFN), celastrol (Cel), Indole-3-carbinol (I3C), -linolenic acid (ALA) or linoleic acid (LA). We propose that the CDDP-WE combination treatment holds the promise for improving curative efficacy for chemo-refractory CRC patients in the future.