Journal of Natural Products

Axenic cultivation of novel bacterial lineages, referred to as "gold standard in microbiology", remains challenging for fastidious or uncultured taxa due the challenges of replicating adequate growth conditions. We isolated strain PM69, a representative of the previously undescribed Bacillota class SHA-98, from a phenyl acid degrading, oligotrophic batch culture. By employing a broad spectrum of (anaerobic) culture techniques, biochemical, physiological, and genomic analyses, we characterised the strain as Thermoaminiphila catenidiffluenda, gen. nov., sp. nov., a thermophilic, strictly anaerobic, bacterium fermenting monosaccharides to acetate. Its motility, biofilm forming capacity, and ecological niche in biogas fermenters and hydrocarbon-associated habitats suggest adaptive strategies for harsh environments exhibiting e.g., high concentrations of aromatic compounds. This description of a new bacterial class not only expands the taxonomic diversity of phylum Bacillota but also provides insights into the metabolic versatility of yet uncultured microorganisms, with implications for carbon cycling and biotechnological applications. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=142 SRC="FIGDIR/small/718153v1_ufig1.gif" ALT="Figure 1"> View larger version (38K): org.highwire.dtl.DTLVardef@109cdd5org.highwire.dtl.DTLVardef@a896faorg.highwire.dtl.DTLVardef@502388org.highwire.dtl.DTLVardef@14ce1fd_HPS_FORMAT_FIGEXP M_FIG C_FIG

Recombinant peptide production was pioneered in the 1970s for the generation of therapeutic peptides, with notable examples including insulin and somatostatin. These early methods required the use of cyanogen bromide (BrCN) for cleavage of the native peptide sequence from a fusion protein. Since that time, while numerous BrCN-dependent peptide methods continue to be reported, the accessibility and cost of site-specific proteases have improved dramatically. These developments have enabled alternative approaches to recombinant peptide generation that obviate the need for BrCN, an environmentally destructive toxin. We recently created an immobilized SUMO protease that can replace BrCN usage in recombinant peptide production workflows by releasing native peptides expressed as part of a SUMO-peptide fusion protein. We have used this approach to generate P113 peptide, the minimal active fragment of the antifungal peptide Histatin 5. In this report, we describe the creation and characterization of this immobilized SUMO protease and its application in the production of experimentally viable quantities of active P113 peptide.

Flavonoids have been widely investigated for their antiviral and anti-inflammatory properties, but their mechanisms of action often remain insufficiently defined. In the present study, high-purity flavonoids were evaluated using an integrated workflow combining molecular docking, LigPlot interaction mapping, surface plasmon resonance (SPR), fluorescence-based TMPRSS2 inhibition assays, and cell-based viability studies. Docking with AutoDock Vina identified Hesperidin as the strongest overall candidate among the compounds evaluated. Hesperidin showed strong active-site engagement with TMPRSS2, including interactions with catalytic residues His296, Asp345, and Ser441, and stable binding within the SARS-CoV-2 main protease (Mpro) pocket. Comparative docking showed weaker or more peripheral interaction patterns for Rutin and moderate Spike binding for Hesperidin and Rutin. Experimental validation demonstrated dose-dependent inhibition of TMPRSS2 activity with an IC50 of 79.1 uM for Hesperidin and 43.5 uM for Hesperetin, while Rutin showed partial inhibition without a defined IC50 in the tested range. In Calu-3 cells, pre-treatment with Hesperidin or Rutin reduced SARS-CoV-2 Spike-induced cytotoxicity by approximately 30 percent without detectable intrinsic toxicity at the concentrations tested. Docking analysis of Hesperidin and Rutin with the SARS-CoV-2 Spike protein revealed moderate interaction patterns involving residues such as Asn343, Ser371, and Val367. Hydrogen bond distances were generally in the range of approximately 2.9-3.3 A, indicating moderate stabilization compared with the stronger active-site interactions observed for Hesperidin in TMPRSS2. The resulting binding poses suggest that these flavonoids can associate with structurally relevant regions of the Spike receptor-binding domain; however, they do not strongly overlap with the key residues required for ACE2 interaction. Rutin, in particular, exhibited a more peripheral and distributed binding mode within the Spike-ACE2 complex, indicating limited potential for direct disruption of the binding interface. In addition to SARS-CoV-2 targets, docking analysis extended to influenza viral proteins revealed moderate interaction of Hesperidin with hemagglutinin (HA) and strong catalytic-pocket binding of Rutin to neuraminidase (NA), involving key residues associated with enzymatic activity. These findings broaden the scope of the study to include influenza viral entry and release mechanisms, supporting a multi-virus, multi-target framework.

Protein synthesis represents an attractive target space for the development of anti-malarials with novel modes of action. Natural-product inhibitors of the eukaryotic 80S ribosome can have potent anti-malarial activity but are often poorly selective due to mammalian cytotoxicity. Blasticidin S (BlaS) is a microbially-produced natural product that broadly inhibits prokaryotic and eukaryotic protein synthesis by binding to the ribosomal peptidyl transferase center. In this study, we explored the potential for improving the anti-malarial potency and selectivity of the blasticidin S scaffold with semi-synthetic analogs that are modified at the C6 and C4 sites. The two best analogs were two orders of magnitude more potent than BlaS against Plasmodium falciparum drug-sensitive and -resistant lines while displaying low cytotoxicity towards mammalian cells. These analogs exhibited improved kinetics of inhibition of protein synthesis in cultured parasites and blocked the development of asexual stages expressing the plasmodial surface anion channel, a transporter required for nutrient acquisition and BlaS uptake. They also exhibited a dramatically improved speed of killing over BlaS. Molecular docking analysis revealed that these analogs are able to form more interactions with the P. falciparum ribosomal peptidyl transferase center than is BlaS, which is consistent with their increased potency. Together, these studies demonstrate the feasibility of generating BlaS analogs with potent anti-malarial activity and provide a roadmap for further development.

The emergence of antimicrobial resistance has been rapid, necessitating the development of alternative therapeutic approaches beyond traditional antibiotics. In this proof-of-concept study, we examined the antibacterial activity of citrate-stabilized, colloidally aggregated silver nanoparticles (AgNPs) against Escherichia coli by combining physicochemical characterization with experimental antibacterial testing The synthesis of silver nanoparticles was done through a modified thermal citrate reduction protocol, and UV-visible spectroscopy, dynamic light scattering (DLS), and zeta potential were used to characterize the nanoparticles. Spectroscopy analysis showed a clear surface plasmon resonance peak at 310-320 nm, indicating the formation of nanoparticles. DLS measurements showed that the dominant hydrodynamic diameter was around 250-270 nm, which is indicative of controlled colloidal aggregation, and near-neutral values of zeta potential indicated steric stabilization of the nanoparticle clusters. Agar tests demonstrated a clear zone of inhibition, and broth cultures showed a lower turbidity and slower bacterial growth with AgNPs. The above findings suggest that nanoparticles that are colloidally aggregated maintain a significant antimicrobial activity even though the surface area is lower than that of monodispersed systems. Mechanistically, the observed antibacterial effect can be explained by a multi-modal effect through direct membrane disruption, localized release of silver ions, and the induction of oxidative stress pathways in bacterial cells. The aggregated form could also help to increase the nanoparticle cell interactions through the provision of multivalent contact points of nanoparticles, and thus the antibacterial efficacy. Controlled colloidal aggregation of AgNPs is a promising approach to the development of effective and possibly more stable antimicrobial agents. These results indicate the possibilities of aggregated nanoparticle systems in fighting drug-resistant pathogens and a basis on future studies of its clinical use.

Powder X-ray diffraction is highly sensitive to sample-delivery conditions, particularly when measurements are performed on platforms originally designed for single-crystal diffraction. In this study, we developed a modified Terasaki-plate-based sample-delivery method for PXRD using a laboratory single-crystal diffractometer implemented with the XtalCheck-S plate-reader module at Turkish Light Source. The method was evaluated against standard loop/pin-based loading and a grease-based Terasaki setup using [4-(2-methoxyphenyl)piperazin-1-yl]methyl}-1,3,4-oxadiazol-2-thiol as a model analyte. While the loop-based method allowed initial PXRD measurements, it provided limited sample volume and insufficient particle statistics. The grease-based plate setup enabled multi-well data collection at a time, but yielded weaker, more diffuse patterns due to increased background noise. Inversely, modification of the Terasaki wells with Kapton tape enabled secure low-volume powder loading, improved diffraction clarity, and supported batch-mode data collection. Comparative search-match and profile-fitting analyses showed that all three loading strategies sampled the same crystalline material, whereas the Kapton-based setup presented the most reliable diffraction profile, with the lowest profile residual (Rp = 9.6%). These findings indicate that this novel sample-delivery method, rather than instrument hardware, can largely improve PXRD performance on an existing in-situ crystallography platform. The Kapton-Terasaki method provides a simple, cost-effective, and practical pipeline for high-throughput PXRD analysis of small powder samples under laboratory conditions.

Microorganisms significantly influence human health, and dysbiosis of the oral microbiome plays a critical role in the development and progression of both oral and systemic diseases. This highlights the urgent need for novel therapeutics targeting specific pathogens. Here, we presented a structure-based pipeline to efficiently identify potential phage-derived periodontal lysins (LysPds) from nearly one million proteins. We predicted the structures of candidate lysins using AlphaFold2 and developed an innovative structure-based similarity network to classify them into distinct clusters, each with unique functional properties. A systematic characterization of 16 representative LysPds from 11 superfamilies revealed that over 90% demonstrated potent antibacterial activity against key periodontal pathogens. Among these, LysPd078 was identified as a promising preclinical drug candidate, effectively reconfiguring microbiome communities while demonstrating significant efficacy and safety in mouse models of periodontitis and calvarial infection. Our findings highlight the effectiveness of structure-based similarity networks in exploring vast protein spaces and underscore the potential of LysPd078 as a targeted modulating agent for the oral microbiome.

Antimicrobial resistance (AMR) has resulted in the need for the development of alternative strategies for combating pathogens and growth promotion of poultry, including the use of plant-derived compounds such as hop (Humulus lupulus) isolates. The present study evaluates the correlation patterns of the biological activity of five major hop isolates (humulone, lupulone, isohumulone, xanthohumol, and isoxanthohumol) against Bacillus subtilis, Micrococcus luteus, and a chicken cell line UMNSAH/DF-1 using a two-dimensional checkerboard assay. Fractional inhibitory concentrations ({sum}FIC) were used to classify interactions as additive, synergistic, or antagonistic, and selectivity indices assessed antibacterial versus cytotoxic effects. On B. subtilis, combinations were predominantly additive, whereas M. luteus, in contrast, showed variable interactions, including also synergistic (humulone + lupulone) and antagonistic combinations (isohumulone + isoxanthohumol), demonstrating the impact of the metabolic resilience of the target organism. Cytotoxicity in UMNSAH/DF-1 cells was largely additive, with synergistic effects observed only for isomerized compounds. Selectivity analysis highlighted humulone-lupulone combinations as most favorable, offering high antibacterial activity with minimal cytotoxicity. These results provide novel insights for selecting hop isolate combinations for the development of phytogenic feed additives (PFAs), emphasizing that both compound composition and target organism physiology critically shape efficacy and safety outcomes.

Chronic obstructive pulmonary disease (COPD), a chronic lung disease, involves complex metabolic disturbances that remain poorly characterized using non-invasive matrices. The metabolic alterations associated with cigarette smoke (CS), one of the major drivers of disease progression in COPD patients, have not been explored in detail. This study primarily aimed to investigate the metabolic signatures in COPD patients categorized into smoker (n=15), ex-smoker (n=11), and non-smoker (n=3) subgroups. Utilizing saliva as a noninvasive sample, we identified 26 metabolites with differential expression in smokers and 31 in ex-smokers. However, no such significant alteration was observed in the non-smokers subgroup. The multivariate analysis distinctly separated the COPD subgroups from healthy controls. Additionally, pathway enrichment analysis revealed perturbations in key metabolic pathways, including unsaturated fatty acid biosynthesis, arginine biosynthesis, the tricarboxylic acid (TCA) cycle, and pyruvate metabolism. Moreover, univariate Random forest analysis identified four metabolites (cyclopentanone, tetradecane 4-methyl, acetophenone, and scyllo-inositol) as potential biomarkers distinguishing COPD subgroups from healthy controls. This study offers novel molecular insights into the association of smoking with disease progression and provides a mechanistic understanding of COPD in different subgroups for better management of the disease. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=163 SRC="FIGDIR/small/717654v1_ufig1.gif" ALT="Figure 1"> View larger version (41K): org.highwire.dtl.DTLVardef@11db4org.highwire.dtl.DTLVardef@1451fb5org.highwire.dtl.DTLVardef@124b62aorg.highwire.dtl.DTLVardef@133872a_HPS_FORMAT_FIGEXP M_FIG C_FIG

Rademikibart (CBP-201) is a human monoclonal antibody with higher binding affinity to IL-4R compared to dupilumab. Dupilumab is a first-generation interleukin-4 receptor alpha (IL-4R) inhibitor for treating IL-4R-dependent inflammatory disorders, including several dermatologic and respiratory conditions. Rademikibart, however, demonstrated better inhibition of STAT6 intracellular signaling in vitro and similar potency in inhibiting both IL-4 induced TARC release and IL-4 induced B cell activation. To further characterize the molecular function of rademikibart and its differentiation from dupilumab, we determined the crystal structure of the rademikibart fragment antigen binding (Fab) bound to IL-4R at 2.71 [A] resolution and compared this to the 2.82 [A] resolution structure of dupilumab Fab bound to IL-4R. The rotation angle between dupilumab and rademikibart bound to IL-4R is 54.88{degrees}. This rotation enables the binding epitopes of rademikibart, but not dupilumab, on IL-4R to overlap more closely with the conserved binding interface naturally utilized by IL-4 and IL-13 cytokines. Molecular dynamics (MD) studies on rademikibart and dupilumab bound to IL-4R examined the stability of the complexes and effects of amino acid mutations on receptor complex formation. MD simulations demonstrated that the third interface loop (residues 145 to 153 in domain 2) of IL-4R interacts directly with rademikibart, which is absent in the dupilumab/IL-4R complex. This finding is confirmed by increased hydrogen bond interactions at the interface between rademikibart and IL-4R, demonstrating superior binding energy for rademikibart. Through analysis of the x-ray crystallography structures, MD-equilibrated structures, and computational point-mutation analysis of rademikibart, we identified residue Y50 and R55 of the light chain and R97, R99, and Y101 of the heavy chain of rademikibart as key residues interacting with IL-4Rs third interface loop. Our data provides a molecular and structural rationale for the enhanced IL-4R inhibition by rademikibart over dupilumab, confirming rademikibart as an optimized second-generation IL-4R inhibitor.

Epitranscriptomics has recently gained significant momentum due to technological advances and translational applications, however, studies on bacterial RNA modifications remain limited. Bacterial RNA remains notoriously prone to degradation and methodologies to investigate the epitranscriptome are challenging. Prior research has shown RNA modifications modulate antimicrobial resistance, virulence and pathogenicity. This research employed CRISPR interference to knock down five known Escherichia coli rRNA modification genes (rlmF, rlmJ, rluD, rsmF and rsmG) in three E. coli strains. These isolates underwent growth curves, proteome analysis and native RNA sequencing CRISPRi adequately silenced the majority of RNA modification genes in E. coli (>80% reduction). Significant growth delays were associated with rlmF, rsmF and rsmG repression. Unique protein pathways corresponding with RNA modification loss were found for rlmJ (TreB, XylF), rluD (CysH, HycB, PutP, TrpB), rsmF (EvgA) and rsmG (OppC). Known rRNA modification sites for rluD ({Psi}) and rsmG (m7G) were detected from analysis of nanopore electrical signal, however, only a weak signal was apparent for m6A (rlmF, rlmJ) and m5C (rsmF) modifications. The inhibition of rRNA modifications resulted in mRNA modification changes including for genes ompC, cspC, dbhA, dbhB and secY. Our work provides an approach for unravelling the epitranscriptome of E. coli and gain insight into its functional role.

Biological nitrification inhibition (BNI) is a plant-mediated process that suppresses nitrification and is widely considered beneficial for reducing nitrous oxide emissions. Here, we show that BNI compounds also inhibit methane oxidation by methanotrophic bacteria, revealing a previously unrecognized trade-off in greenhouse gas regulation. Across soil bioreactor systems and pure cultures of both Type I and Type II methanotrophs, BNI compounds consistently suppressed methane oxidation activity. Kinetic analyses indicated an uncompetitive-like inhibition pattern, characterized by concurrent decreases in Vmax and Km, while reversibility assays showed that inhibition was not associated with loss of cellular viability. Experiments under copper-replete and copper-depleted conditions further showed that inhibition is predominantly associated with particulate methane monooxygenase (pMMO). Transcriptomic analyses demonstrated compound-specific responses, including suppression of methane oxidation pathways and differential regulation of stress-associated genes. These findings suggest that BNI-mediated inhibition of methane oxidation may offset reductions in nitrous oxide emissions, with implications for predicting net greenhouse gas fluxes in agricultural and wetland ecosystems. Incorporating BNI effects into biogeochemical models will be critical for accurately evaluating their role in the global methane budget.

Pantothenamides (PanAms) comprise a promising class of antimalarial compounds that kill asexual blood-stage Plasmodium falciparum parasites and block transmission. Intriguingly, the most advanced PanAm in drug development, MMV693183, is approximately 100 times more potent against female gametocytes than males. We hypothesized that this specificity is explained by a difference in PanAm uptake, which we studied using a PanAm-based photoaffinity labelling (PAL) probe. We successfully synthesized a probe that competed with MMV693183 in drug sensitivity assays, while the probe did not display high potency by itself. We observed no significant difference in median fluorophore-labelled probe signal intensity between male and female gametocytes, although there might be a difference in subcellular localization of the probe between the sexes. By combining PAL with affinity purification and mass spectrometry, we were not able to identify novel candidate PanAm transporters. We conclude that PAL provides evidence that differences in PanAm uptake do not underly differences in PanAm sensitivity between the gametocyte sexes.

Irritable Bowel Syndrome (IBS) affects a substantial proportion of university students, yet its factors remain incompletely characterised in South Asian populations. We reanalysed a publicly available dataset of 550 Bangladeshi students from Hasan et al. (2025), conducting a data audit that identified implausible records, including males reporting menstrual symptoms, and reduced the analytic sample to 506 observations. Using Explainable Boosting Machines (EBMs), which capture non-linear effects and pairwise interactions without sacrificing interpretability, we found that psychological distress, elevated BMI and academic dissatisfaction were the strongest predictors of IBS (mean AUC = 0.852 across 100 stratified train-test splits). Critically, several findings diverged from the original logistic regression analysis. Physical activity showed a non-linear risk pattern only at high intensity, the association with gender was substantially weaker when we accounted for metabolic and psychological factors as well and malnourishment does not have a strong an impact as in the original study. These divergences likely arise because the machine-learning model captures non-linear effects and interactions that were not represented in the original regression specification. Our findings underscore the value of reanalysing existing datasets with methods suited to capturing complexity and highlight data quality verification as a necessary step in the secondary analysis.

BackgroundPatent ductus arteriosus (PDA) is a common and potentially serious cardiovascular condition in preterm infants, particularly those with low gestational age and birth weight. Its management remains controversial due to variability in screening, diagnostic criteria, and treatment strategies. This study aimed to evaluate risk factors, outcomes, and management strategies for PDA in preterm infants, and to identify predictors of clinical and echocardiographic response to therapy. MethodsWe conducted a retrospective cohort study over a 4-year period (2016-2019) in the neonatal intensive care unit (NICU) of a tertiary care center. All consecutive preterm infants admitted during the study period were eligible. Infants with echocardiographically confirmed PDA who received pharmacological treatment with intravenous paracetamol or ibuprofen were included in the analysis. Missing data were minimal and handled using available-case analysis. Statistical analyses included descriptive statistics, Pearsons chi-square test, and multivariable logistic regression. ResultsAmong 2154 preterm infants admitted to the NICU, 60 were diagnosed with PDA (incidence : 2.8%). The mean gestational age was 29 {+/-} 2.6 weeks, and the median birth weight was 1200 g. Respiratory distress occurred in 95% of cases, mainly due to hyaline membrane disease (86.7%). PDA was symptomatic in 80% of infants. First-line treatment resulted in clinical improvement in 77% and ductal closure in 83.3% of cases, most within 3 days. Predictors of successful closure included gestational age [&ge;] 28 weeks (OR = 5.9; 95% CI : 1.7-20.2) and antenatal corticosteroid exposure (OR = 1.2; 95% CI : 1.0-1.6). Overall mortality was 35% and was significantly higher in infants < 28 weeks (OR = 5.0; 95% CI : 2.4-10.3). Clinical improvement (OR = 3.7) and echocardiographic closure (OR = 4.5) after first-line treatment were associated with reduced mortality. ConclusionsPDA in preterm infants is associated with substantial morbidity and mortality, particularly in those born before 28 weeks of gestation. Early diagnosis, antenatal corticosteroid exposure, and timely pharmacological treatment may improve outcomes. Systematic echocardiographic screening in high-risk neonates should be considered.

ImportanceGuideline-concordant care for young children with attention-deficit/hyperactivity disorder (ADHD) includes recommending parent training in behavior management (PTBM) as first-line treatment. However, assessing guideline adherence through manual chart review is time-consuming and costly, limiting scalable and timely quality-of-care measurement. ObjectiveTo evaluate the accuracy and explainability of large language models (LLMs) in identifying PTBM recommendations in pediatric electronic health record (EHR) notes as a scalable alternative to manual chart review. Design, Setting, and ParticipantsThis retrospective cohort study was conducted in a community-based pediatric healthcare network in California consisting of 27 primary care clinics. The study cohort included children aged 4-6 years with [&ge;] 2 primary care visits between 2020-2024 and ICD-10 diagnoses of ADHD or ADHD symptoms (n=542 patients). Clinical notes from the first ADHD-related visit were included. A stratified subset of 122 notes, including all cases with model disagreement, was manually annotated to assess model performance in identifying PTBM recommendations and rank model explanations. ExposuresAssessment and plan sections of clinical notes were analyzed using three generative large language models (Claude-3.5, GPT-4o, and LLaMA-3.3-70B) to identify the presence of PTBM recommendations and generate explanatory rationales and documentation evidence. Main Outcomes and MeasuresModel performance in identifying PTBM recommendations (measured by sensitivity, positive predictive value (PPV), and F1-score) and qualitative explainability ratings of model-generated rationales (based on the QUEST framework). ResultsAll three models demonstrated high performance compared to expert chart review. Claude-3.5 showed balanced performance (sensitivity=0.89, PPV=0.95, and F1-score=0.92) and ranked highest in explainability. LLaMA3.3-70B achieved sensitivity=0.91, PPV=0.89, and F1-score=0.90, ranking second for explainability. GPT-4o had the highest PPV [0.97] but lowest sensitivity [0.82], with an F1-score of 0.89 and the lowest explainability ranking. Based on classifications from the best-performing model, Claude-3.5, 26.4% (143/542) of patients had documented PTBM recommendations at their first ADHD-related visit. Conclusions and RelevanceLLMs can accurately extract guideline-concordant clinician recommendations for non-pharmacological ADHD treatment from unstructured clinical notes while providing clear explanations and supporting evidence. Evaluating model explainability as part of LLM implementation for medical chart review tasks can promote transparent and scalable solutions for quality-of-care measurement.

Background: Lateral lymph node metastasis (LLNM) is associated with poor prognosis in patients with rectal cancer and may influence the indication for lateral lymph node dissection. Accurate preoperative identification of LLNM remains challenging. This study aimed to develop and internally validate a clinicoradiological model for preoperative prediction of LLNM in rectal cancer. Methods A retrospective cohort of 64 patients undergoing lateral lymph node dissection (LLND) for rectal cancer was analysed; 21 (32.8%) had pathological lateral lymph node metastasis (LLNM). A prespecified preoperative clinicoradiological model was fitted using penalised logistic regression with L2 regularisation (ridge), incorporating MRI-measured lateral lymph node short-axis diameter (LLN-SAD), dichotomised clinical T stage (T3-4 vs T1-2), dichotomised clinical N stage (N+ vs N0), and log(CA19-9+1). Model performance was evaluated using the area under the receiver operating characteristic curve (AUC), calibration analysis, and bootstrap internal validation. Results The model showed good discrimination (AUC 0.914), with an optimism-corrected AUC of 0.887 on bootstrap validation. Calibration remained acceptable after optimism correction (calibration intercept -0.127; slope 1.045). Decision curve analysis suggested net benefit across clinically relevant threshold probabilities, particularly between 0.10 and 0.30. The model was implemented as a web-based calculator to facilitate clinical use. Conclusion This clinicoradiological model showed good discrimination, acceptable calibration, and potential clinical utility for preoperative assessment of LLNM risk in rectal cancer. It may assist individualized risk stratification and treatment planning, although external validation is required before routine clinical implementation.

Objective Cerebral palsy (CP) affects approximately 1 million Americans and 18 million individuals worldwide, yet contemporary US epidemiologic data remains limited. We aimed to use Cerebral Palsy Research Network (CPRN) clinical registry to describe demographics and clinical characteristics of individuals with CP across the US and determine associations with gross motor function and genetic etiology. Methods Registry subjects were included if they had clinician-confirmed CP and prospectively entered data for Gross Motor Function Classification System (GMFCS) Level, gestational age, genetic etiology, CP distribution, and tone/movement types. Logistic regression was used to determine which of these variables plus race, sex, ethnicity, and age were associated with GMFCS level and genetic etiology. Results A total of 9,756 children and adults with CP from 22 CPRN sites met inclusion criteria. Participants were predominantly White (73.0%), male (57.3%), non-Hispanic (87.8%), and younger than 18 years (73.7%). Most were classified as GMFCS levels I-III (55.6%), born preterm (52.8%), had spasticity (83.8%), and had quadriplegia (41.9%); 12.2% were identified as having a genetic etiology. Tone/movement types, CP distribution, and gestational age were significantly associated with both GMFCS level and genetic etiology (p<0.001). Compared to White individuals, Black individuals were more likely to have greater gross motor impairment (p<0.001). Conclusion In this large US cohort, clinical and demographic factors, including race, were associated with gross motor function and genetic etiology in CP. These findings highlight persistent disparities and demonstrate the value of a national clinical registry for informing prognostication, quality improvement efforts, and targeted genetic testing strategies.

Understanding depressive symptoms dynamics and their determinants is crucial for designing effective mental health support initiatives. This study compared two methods for describing youth depressive symptoms trajectories and investigated associations of early-life factors (maternal education, maternal perinatal depression, domestic violence, physical, emotional, or sexual abuse, bullying victimisation, psychiatric disorder) with trajectory features. Prospective data from 8,264 mostly White European participants (54% female), including self-reported Short Moods and Feelings Questionnaires on ten occasions between 10-25 years, were used. Trajectories were summarised using functional principal component analysis (FPCA) and P-splines linear mixed-effect (PLME) models. Estimated derivatives were used to obtain magnitude and age of peak symptoms and peak symptoms velocity. Both methods performed comparably, but PLME models tended to over-smooth trajectories. Peak symptoms and peak velocity were higher and occurred >1 year earlier in females than males. All early-life factors were associated with higher peak symptoms, and most associated with higher and earlier peak velocity. Abuse and bullying additionally associated with earlier age of peak symptoms. FPCA is a useful alternative for characterising depressive symptoms trajectories and informing time-sensitive preventative measures to reduce impact of depression before symptoms reach their peak. Early-life stressors may accelerate timeline and intensity of symptoms escalation during adolescence. Lay summaryUnderstanding development of depressive symptoms and factors shaping them is crucial for designing effective mental health support initiatives. This study used data from over 8,000 young people regularly followed up from before birth to compare two cutting-edge methods for describing depressive symptoms trajectories and examined how known risk factors for adulthood depression relate to the severity and rate of change of depressive symptoms in adolescence. We found that both methods performed well and that the peaks in depressive symptoms and their rate of change were, on average, higher and occurred over a year earlier in females than males. Our findings additionally suggest that early-life stressors (e.g., abuse, bullying) may accelerate the development of depression, highlighting the importance of early prevention.

Background: Preterm births contribute to approximately 35% of neonatal deaths globally, with an estimated 13.4 million infants born prematurely each year. Despite this substantial burden, limited evidence exists on time to discharge and its determinants among preterm neonates admitted to Neonatal Intensive Care Units (NICUs), particularly in rural Ugandan settings. This study aimed to investigate time to discharge and associated factors among preterm neonates admitted to Kiwoko Hospital in Nakaseke District, Uganda. Methods: A retrospective cohort study was conducted using secondary data from Kiwoko Hospital on preterm neonates admitted to the Neonatal Intensive Care Unit (NICU) between 2020 and 2021 (n = 847). The cumulative incidence function was used to estimate the probability of discharge within 28 days of admission, accounting for competing events. A Fine and Gray sub-distribution hazard regression model was fitted to identify factors associated with time to discharge. Results: Of the 847 preterm admissions, 70.1% were discharged alive within 28 days. The median time to discharge was 14 days. The cumulative incidence of discharge by 28 days was 68%, accounting for competing events. During follow-up, 165 neonates did not complete the 28-day period, including 88 deaths. Factors significantly associated with time to discharge included place of delivery (SHR: 0.62; 95% CI: 0.53-0.73; p<0.001), maternal residence in other districts (SHR: 0.69; 95% CI: 0.48-0.99; p=0.044), extreme preterm (SHR: 0.05; 95% CI: 0.03-0.09; p<0.001), very preterm (SHR: 0.18; 95% CI: 0.14-0.25; p<0.001), moderate preterm (SHR: 0.59; 95% CI: 0.46-0.76; p<0.001), triplet births (SHR: 0.40; 95% CI: 0.23-0.68; p=0.001), 2-4 ANC visits (SHR: 0.70; 95% CI: 0.56-0.87; p=0.002), <=1 ANC visit (SHR: 0.64; 95% CI: 0.49-0.85; p=0.002), respiratory distress syndrome (SHR: 0.64; 95% CI: 0.48-0.74; p<0.001), and birth trauma (SHR: 2.62; 95% CI: 1.60-4.29; p<0.001). Conclusions: Respiratory distress syndrome, fewer antenatal care visits, out-of-district residence, and higher degrees of prematurity were associated with prolonged time to discharge among preterm neonates. Strengthening antenatal care utilization and improving access to quality neonatal care in underserved areas may enhance discharge outcomes.