Molecular tumor boards (MTBs) integrate clinical, pathological, and molecular data to prioritize therapy options. Because MTB-guided treatment starts at varying time points after MTB discussion, vulnerabilities to immortal time bias occur if treatment delay is ignored. Retrospective survival analysis of 949 consecutive individuals discussed at the University Cancer Center Hamburg MTB (2016-2022). Index date (t0) was the first MTB discussion. Therapy initiation was modeled as a time-dependent exposure using Mantel-Byar methods and visualized with Simon-Makuch plots. Therapy strata were priority 1 targeted, priority 2 targeted, and non-targeted therapy. Among solid tumors (n = 854), targeted therapy was implemented in 24% and was off-label in 51%. ORR/DCR were 30.8%/61% for priority 1, 11.4%/36.4% for priority 2, and 37.5%/52.1% for non-targeted therapy (P = .017). The progression-free survival ratio (MTB-directed therapy vs immediately preceding line) exceeded 1.3 in 63% of targeted therapy recipients. From the MTB date, median survivalMTB was 16.5 months (priority 1), 15.5 months (priority 2), and 11.0 months (non-targeted). In Mantel-Byar analysis, progression-free survivalSimonMakuch favored priority 1 (P = .002) with 1-year rates of 28%, 12%, and 15% for priority 1, priority 2, and non-targeted therapy, respectively. One-year overall survivalSimonMakuch was 52%, 34%, and 42%. In a time-dependent Cox model, priority 1 was associated with lower risk of death versus no targeted post-MTB therapy (hazard ratio 0.80, 95% confidence interval 0.64-0.99, P = .038). After correction for immortal time bias, implementation of the top-priority MTB recommendation was associated with improved disease control and survival signals, although confounding and selection effects remain key limitations.

Previous research has shown that observers can predict the target object of a grasping action from early hand preshaping cues. However, critical questions remain unexplored: how predictions adapt to the available kinematic information and evolve throughout the movement timeline. We address these fundamental gaps by combining kinematic analysis with machine-learning approaches. Using motion capture technology, we recorded reach-to-grasp actions toward large and small objects and had participants predict target size from hand kinematics at varying time points. Our analysis revealed that prediction performance not only evolved with increasing information but, crucially, differed significantly between target size choices. To provide insight into the participants' performance, we developed a comparative framework using two distinct machine learning models: Support Vector Machines modeling kinematic information and convolutional neural network-recurrent neural networks extracting visual patterns. This comparison indicates that predicting the target objects of observed actions adapts to the available kinematic information depending on the target object, with prediction changing over time accordingly. These findings advance our understanding of action prediction and have significant implications for social cognition and human-machine interaction.

Postparathyroidectomy Bone Density Changes in Patients With Biochemically Mild Primary Hyperparathyroidism.

There is limited research on women's perception of risk regarding pregnancy after caesarean. Interpregnancy interval alters risk in the next pregnancy and birth following a caesarean. Interpregnancy interval is a potentially modifiable risk that health care providers need to consider when counselling women about future pregnancy plans. What is women's experience of decision making regarding interpregnancy interval after caesarean birth? Twelve women participated in in-depth interviews; these were analysed thematically assisted by the use of NViVO software. The overarching theme identified was "Finding my way through the confusion to make informed decisions". Women are receiving confusing biased information delivered at varying time points. This is not meeting women's information needs leading them to seek knowledge through other sources, including online peers, to make sense of the confusion. They gather their own knowledge in order to advocate for themselves, make pregnancy spacing decisions that fit with their expectations and maximise the chances of having the birth they want. Consistent evidence based information is lacking in the current maternity care environment regarding interpregnancy interval. This creates barriers to making informed decisions for next pregnancy and birth planning. Women are looking for evidence based information to make individualised decisions. This requires women to seek knowledge outside of the maternity care system to empower themselves to navigate the system and make decisions rather than following prescriptive directives. There is a need to provide evidence based pregnancy planning resources to women that facilitate informed shared decision making.

Diurnal changes of cerebrospinal fluid and global signal coupling.

When bacteria are treated with multiple antibiotics simultaneously, resistance is highly unlikely to evolve. In contrast, resistance against multiple phages frequently arises during therapy. Why does resistance against multi-phage cocktails evolve so easily? Using a mathematical model, we show how the bacterial evolutionary dynamics and phage replicative dynamics uniquely intertwine, facilitating the rapid evolution of multi-phage resistance. As different phages replicate and become inhibitory at varying time points, bacteria can sequentially acquire resistance rather than simultaneously - increasing the chance of multi-resistance by orders of magnitude. We predict and experimentally verify a regime where multi-phage resistance is robustly prevented. Our findings provide a framework for the rational design of phage cocktails to curtail resistance development. Resistance can be minimized by reducing the dose of the most potent phages or by using phages with longer latent periods, as this helps synchronize multi-phage selection.

The restricted regenerative potential of adult hearts poses a significant barrier to effective repair following injury. In contrast to numerous vertebrates, mammalian hearts exhibit only transient neonatal renewal capacity during the initial days of life. Beyond cardiomyocytes, understanding the diverse compositions of non-cardiomyocytes (non-CMs) is imperative for maintaining heart microenvironment homeostasis during neonatal heart regeneration. Here, we conduct single-cell ATAC sequencing on neonatal hearts at varying time points post-apical resection to profile the epigenetic landscape. Intriguingly, fibroblasts and endothelial cells, as the most abundant populations in the heart, exhibit the most dynamic chromatin remodeling upon injury. Furthermore, we reveal CEBPD and AP-1 family transcriptional factors as pivotal trans-regulators orchestrating these alterations, governing beneficial fibroblast activation and endothelial cell angiogenesis crucial for cardiac regeneration, respectively. Collectively, our study delineates the cellular identity of non-CMs at the epigenome level using single-cell approaches, offering insights into cell type-targeted interventions for heart regeneration.

Completion of follow-up colonoscopy after a positive stool-based test varies across health settings. Colonoscopy performed > 6months following a positive stool test is associated with adverse colorectal cancer outcomes. This retrospective study included 701 patients aged 45-75 in an urban, integrated health system with a positive stool test between February 1, 2022, and January 31, 2023. We examined rates of timely (i.e., within 180days) follow-up colonoscopy and at varying time points (90days, 365days, and any time during our follow-up period). Multivariable Cox proportional hazard models examined factors associated with timely colonoscopy, and a Pareto analysis identified barriers to timely completion. The median age of this cohort was 64years (IQR 56-70); 52.1% were female, 48.2% were non-Hispanic White, and 52.4% had a Charlson Comorbidity Index score ≥ 3. The rate of timely follow-up colonoscopy was 59.6%, with rates of 44.5% and 68.5% at 90days and 365days, respectively. In Cox models, patient outreach increased timely colonoscopy by 52% (HR 1.52, 95% CI 1.21-1.91), while Direct Access patients were less likely to complete timely colonoscopy (HR 0.59, 95% CI 0.41-0.86); no associations were observed with sociodemographic factors. The most common barriers to timely colonoscopy were (1) lack of gastroenterology clinic visit, (2) patient refusal, and (3) colonoscopy no-show or cancellation. The rate of timely, follow-up colonoscopy in this older, sicker population is suboptimal. System-level factors impact timely completion. A Pareto analysis reveals multiple elements that contribute to delays in colonoscopy and can inform interventions.

Abstract Background Dysregulated STING signalling in intestinal and colonic tissue is associated with dysbiosis and increased susceptibility to enteric pathogens, however, the correlation between the cGAS-STING pathway and IBD appears to be context dependant and requires further investigation. Human adenovirus are non-enveloped dsDNA viruses classified into 7 distinct species. Species F includes both type 40 (Ad40) and type 41 (Ad41); enteric pathogens that specifically infect the intestinal epithelium and are the third leading cause of infantile gastroenteritis. Adenovirus infection has been shown to disrupt the host microbiome disrupting intestinal homeostasis, however, research into the association of viral infection and STING activation in the intestinal epithelium remains poorly characterized. Aims I hypothesize that epithelial intrinsic STING signalling serves a critically protective role in the host defence response, with dysregulation causing increased susceptibility to viral infection. Aim 1 is to characterize the physiological effect of epithelial intrinsic STING signalling to intestinal homeostasis. Aim 2 is to then investigate the implications of the STING-mediated response to enteric adenovirus infection. Methods Through our existing collaboration with the Canadian National Organoid Network (CNON) and existing library of patient-derived organoid samples, I utilize ileal and duodenal organoids to serve as a primary human model. To elucidate if the cGAS-STING pathway serves as the primary line of defence against enteric adenovirus, I infected human duodenal and ileal organoids with Ad41 to study the host response to infection through western blot, RT-qPCR, and confocal microscopy. Results Human duodenal organoids possess a functional cGAS-STING pathway, activated by diABZI. Despite robust STING activation by agonists, infection of organoid with Ad41 did not trigger STING phosphorylation, even at varying time points or high viral doses. Infection was confirmed by expression of viral transcripts, suggesting Ad41 evades STING-mediated antiviral signaling in intestinal epithelial cells. Conclusions Previous studies on adenovirus sensing by cGAS-STING have been limited to cancer or immune cells infected with the respiratory Ad5 strain. My findings indicate that enteric adenovirus evades cGAS-STING detection in human intestinal epithelium, revealing its ability to exploit this tissue as a replicative niche. Funding Agencies CCC, CIHRNew Frontiers in Research Fund

Calcitonin gene-related peptide (CGRP) is thought to be a key player in the pathogenesis of migraine, but there is a fundamental mystery in that the known neuronal actions of CGRP do not account for how it causes pain. We now report the first finding of CGRP-induced nociceptive neuronal activation using a novel method of intra-carotid infusion to achieve a more targeted delivery to cranial tissues. Single-unit recordings were performed in anesthetized rats to measure CGRP effects on first- and second-order trigeminovascular neurons. CGRP was administered via intra-carotid infusion. Neuronal activation and sensitization were assessed by spontaneous firing rates and responses to mechanical stimulation of dural and facial receptive fields. Lidocaine was applied locally to the dura or trigeminal ganglion at varying time points to determine the peripheral contribution to CGRP-induced activity. Intra-carotid CGRP infusion (5 µg/kg/min, 20 min) activated 62% of Aδ-fibers and 56% of C-fibers, with significant increases in firing rates beginning within the first 30 minutes for Aδ-fibers and after one hour for C-fibers. It also activated 75% of central trigeminovascular neurons, significantly increasing spontaneous firing and sensitizing dural and facial receptive fields. Similar effects were produced by CGRP injection into the trigeminal ganglion. These effects of CGRP were impeded by local anesthetic blockade of the dura or trigeminal ganglion before but not 1 hour after CGRP infusion. No significant sex differences were found in baseline firing or in the magnitude and timing of CGRP-induced responses across all neuron types. These finding provide the first evidence of peripheral nociceptive neuronal activation by CGRP, with a site of action in the meninges, and support a rationale for early, peripherally acting CGRP-targeted migraine treatments.

Tissue-bone homeostasis manipulation (TBHM) has been proven effective for knee osteoarthritis (KOA), but its optimal timing and underlying mechanisms remain unclear. Melatonin serves as a key biomarker of circadian rhythm, while electroencephalography (EEG) evaluates pain-related central mechanisms. This study investigated the efficacy of TBHM at different time points based on circadian principles and explored potential central mechanisms using EEG. In this 4-week randomized controlled single-blind trial, 88 KOA patients were randomized into four groups: group A (TBHM at 8 a.m.), group B (TBHM at 1 p.m.), group C (TBHM at 6 p.m.), and group D (joint mobilization). Interventions were administered once daily (20min/session, 5 days/week). Primary outcome was Visual Analog Scale (VAS) for pain; secondary outcomes were resting-state EEG and Hamilton Anxiety Rating Scale. Salivary melatonin levels were measured to explore circadian mechanisms. Assessments were conducted at baseline and after 4 weeks. Statistical analyses employed two-way repeated-measures ANOVA. Of the 88 patients randomized, 82 completed the study. After 4 weeks, all groups showed reduced VAS scores and increased melatonin levels. Post-treatment, group C exhibited significantly lower VAS scores than group B (FDR adjusted P = 0.023), and group B had significantly lower VAS scores than group A (FDR adjusted P = 0.037). Although group A showed lower scores than group D, the difference was not statistically significant (FDR adjusted P > 0.05). Melatonin levels increased significantly in the three TBHM groups after treatment. group C was demonstrated significantly higher melatonin levels than group B (FDR adjusted P < 0.001), group B was significantly higher than group A (FDR adjusted P = 0.007), and group A was higher than group D (FDR adjusted P = 0.026). After treatment, a decrease in β band activity and an increase in θ band activity were observed in the frontal and central regions of groups C and B in EEG analysis, but there was no significant difference (FDR adjusted P > 0.05). The TBHM at all time points can better improve the pain of KOA patients than joint mobilization, regulate cortical electrical activity, and increase the secretion of melatonin at night. ChiCTR2400080820. Registered on Feb.07,2024.

Abstract Introduction Limited data exist on semen parameters post vasectomy reversal. Specifically, no studies have rigorously evaluated the likelihood of sperm return following discovery of azoospermia at some time point post vasectomy reversal. Objective To evaluate the rate of return of sperm among men who experience an azoospermia result post vasectomy-reversal. Methods A large, prospective dataset has been maintained of men undergoing vasectomy reversals at a single center from 2019 to the present. Men who had at least two post-operative semen analyses were included for review. The data were then analyzed in a manner to identify those who experienced a 0 count at varying time points and then either went on to continue to have 0’s beyond that point or who experienced a return of sperm. Results A total of 4,335 men underwent a vasectomy reversal since 2019, with 3,517 having semen analysis data available for review. Overall demographics demonstrated a mean age of 41, partner age 33, median time since vasectomy of 6 years, 93% first-time cases, and distribution of repair as noted: VV/VV (81%), VV/EV (12%), EV/EV (5%), VV/x (1%), EV/x (&amp;lt;1%), x/x (0%). Of men who had 0 sperm at varying time points, the percentages of those who experienced a return of sperm are noted as follows: 0 sperm at &amp;lt;2 months (52% experienced a return at some point, n=467 used in calculation), 0 at 2-4 months (36% return, n=250), 0 at 5-7 months (17% return, n=226), 0 at 8-10 months (20% return, n=147), 0 at 11-18 months (11% return, n=111). The data are limited by the fact that relatively few men continued to send in semen analysis samples beyond 12 months (n=481 with at least one result &amp;gt;12 months and n=160 with results &amp;gt;24 months). Conclusions Men who experience azoospermia post-vasectomy reversal may experience a return of sperm over time, with the likelihood for return declining based on the duration of persistence of 0 sperm. Disclosure Any of the authors act as a consultant, employee or shareholder of an industry for: PathRight medical

Akkermansia muciniphila (Akk), a mucin-degrading bacterium residing in the human gut, plays a pivotal role in intestinal health. This study investigated its temporal dynamics, strain-level diversity, and cross-regional transmission using longitudinal metagenomic data from the Chinese Microbiome Project (CMP). We observed significant fluctuations in Akk relative abundance across 52 time points in 7 healthy individuals, with detection rates of 56.9% (16S rRNA gene sequencing) and 33.3% (whole-genome sequencing, WGS). Notably, "short-term blooms"–-rapid increases followed by declines in relative abundance–-were identified in multiple subjects. Genomic analysis of 39 Akkermansia metagenome-assembled genomes (MAGs), combined with 89 publicly available strains with complete genome, revealed phylogenetically distinct clusters (average nucleotide identity, ANI < 98% between clusters). Strikingly, individuals harbored different clusters at varying time points (e.g., AmII replaced by AmIb and later AmIa in subject P4), suggesting strain replacement and recurrent colonization. Furthermore, high-similarity strains (ANI > 99%) were shared between individuals with close contact (e.g., cohabiting subjects P2 and P4) and across geographically distant regions (China, South Korea, and the United States), implicating human-mediated or environmental transmission pathways. These findings underscore the dynamic nature of Akk within the gut microbiota and highlight the need to explore factors driving its colonization, strain competition, and ecological dissemination.Supplementary InformationThe online version contains supplementary material available at 10.1186/s13568-025-01982-7.

Impact of adolescent ethanol binge on serotonin signaling and pain sensitivity post-withdrawal.

Background: Prediction of sudden cardiac arrest (SCA) from the ECG is of great importance, but is complicated by the uncertainty introduced by dynamic remodeling of the ECG over time. Traditional machine learning models trained on ECG snapshots have not captured this dynamic remodeling. Hypothesis: Artificial intelligence (AI)- models trained on temporal ECG feature sequences will reveal dynamic biomarkers of cardiac arrest. . Methods: Our registry consisted of 108,704 12-lead ECG recordings from 2,837 patients (age=61 ± 14 years, 34% female, 46% white) who experienced cardiac arrest within 1 year. Routine ECG features (e.g. P-duration, QRS-duration, QT-interval) were indexed by “time to arrest”. Training, test and validation cohorts were created using patient-stratified splitting, and training data was augmented to make the transformer model robust against noisy timestamps and data sparsity. We calculated the importance of features dynamically over time using Harrell’s Concordance Index (C-Index) and global SHAP values at multiple prediction horizons (30-360 days). (Fig 1) Results: In the training cohort, SCA prediction performance peaked at 60 days before arrest (C-index = 0.89) and tapered monotonically thereafter. In the hold-out test cohort, SCA prediction peaked at 120 days (C-index = 0.78) and fell only slightly to 0.76 at 365 days (Fig 2). Features most important for long-term prediction were the RR-interval (importance ~50%), with an emergence of QT-interval, P-duration and PR-interval as additional co-predictors at timepoints closer to the SCA event (cumulative importance ~ 32%) (Fig 3). Conclusion: ECG-features’ vary in their importance to predicting SCA over a one-year period. Using time-aware AI-transformer models, RR interval was the strongest predictor from 1 year preceding the event, with other features emerging as the event draws closer. This study shows the importance of using repeated 12-lead ECGs in at-risk patients, and combining them into dynamic-predictive indices.

Long-read whole genome sequencing improves clinical management in acute leukemia

BackgroundEarly prediction of upper limb recovery is important to optimise rehabilitation and inform patients but remains challenging due to inter-individual variability. This study aims to (1) develop and validate a machine learning model to predict arm-hand capacity at six months post-stroke using clinical variables from the first week; (2) compare its performance to a mixed-effects model; and (3) co-design a user-friendly output visualisation with clinician input.MethodsFrom data of 451 first-ever ischemic stroke patients, we selected total Action Research Arm Test score (ARAT), shoulder abduction, and finger extension as predictors. An XGBoost model was trained on these variables measured at varying time points within the first five months, using 5-fold, 5-repeat cross-validation. We employed bootstrap aggregation to obtain generalisable predictions and prediction intervals to quantify uncertainty. The model's performance was validated on a hold-out set and compared against a mixed-effects model using median absolute error (MedAE).Results The XGBoost model achieved a MedAE of 4.2 points (IQR = [1.2, 12.6]) on the ARAT when applied at seven days post-stroke, compared to 13.7 points (IQR = [4.6, 27.8]) for the mixed-effects model in the same patients.ConclusionOur model provides significantly more accurate predictions of upper limb recovery, with a 69% error reduction compared to the mixed-effects model. Its ease of use, interpretability, and use of routinely collected clinical data make it suitable for digital clinical workflows. Future research could validate the model in larger, more recent cohorts and explore integrating neuroimaging and temporal features.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12984-025-01743-4.

ABSTRACT Previous research has established that observers can predict action targets through hand preshaping. However, two critical questions remain unexplored: how predictions adapt to the available kinematic information and evolve throughout the movement timeline. We address these fundamental gaps by combining kinematic analysis with machine-learning approaches that differentiate between motor and visual cues. Using motion capture technology, we recorded reach-to-grasp actions toward large and small objects and had participants predict target size from hand kinematics at varying time points. Our analysis revealed that prediction performance not only evolved with increasing kinematic information but, crucially, differed significantly between target size choices. To provide insight into the underlying processes, we developed a comparative framework using two distinct machine learning approaches: Support Vector Machines (SVM) modeling kinematic information and CNN-RNN networks extracting visual patterns. The stronger alignment between human performance and SVM predictions offers empirical evidence that kinematic cues, rather than visual patterns, mostly guide target prediction. These findings advance our understanding of action prediction and have significant implications for social cognition and human-machine interaction. PUBLIC SIGNIFICANCE STATEMENT Understanding others’ intentions by observing their hand movements is crucial for social interaction, from passing objects to coordinating complex tasks. This study reveals that people use different cues to predict whether someone is reaching for a large versus a small object from the earliest stages of hand movement. By comparing human performance with Artificial Intelligence models, we found that people primarily rely on motor cues to make these predictions. These insights could improve rehabilitation techniques for individuals with social interaction difficulties and enhance the design of intuitive robotic assistants.

ABSTRACTUnraveling the nanoscale distribution of small molecules in cells is of central importance for the understanding of cellular functions and the development of drugs. However, particularly the visualization of lipids such as cholesterol—a central compound of cell membranes—with high spatial resolution remains challenging because they cannot be efficiently immobilized for super‐resolution microscopy investigations. Here we developed an azido‐ and amino‐modified cholesterol probe that can be efficiently fixed and labeled with fluorophores by click chemistry. In combination with expansion microscopy, its cellular localization and interaction with other cellular proteins can be precisely determined in fixed cells at varying time points after addition. Our approach allows us to detect the endocytic pathway of cholesterol with unprecedented spatial resolution and shows that cholesterol is efficiently ingested in endocytic vesicles and accumulates as cholesterol aggregates with an average size of ∼37 nm in late endosomes and lysosomes, respectively.

8038 Background: Osimertinib has become the standard adjuvant treatment for patients (pts) with surgically resected, early-stage, EGFR -mutated non-small cell lung cancer (NSCLC), after the results of the ADAURA trial. However, the temporal distribution of EGFR mutations (mut) and its relation with recurrence patterns in this population have not been established yet. Aim of the study is to describe the prevalence of circulating tumor DNA and EGFR mut at different timepoints in early-stage NSCLC and the correlation between their presence and prognosis. Methods: This is a single center study conducted at the Vall d’Hebron Institute of Oncology, including consecutive pts with surgically resected, stage I-III NSCLC harbouring EGFR mut from 2008 to 2024. Next Generation Sequencing (NGS) with ONCOMINE panel was performed on tissue samples, archival when surgery pre-dated the start of the study. NGS on plasma samples was performed using Guardant360 panel at timepoints: 1, 3 and 6 months after surgery and at recurrence. For pts receiving adjuvant osimertinib, additional plasma samples were collected before drug initiation and during treatment. Pts were referred for genetic consultation if NGS on tissue samples detected a TP53 mut with a variant allele frequency (VAF) &gt;30%, or if NGS on plasma identified TP53 or BRCA mut with a VAF &gt;20%, or a basal T790M mutation. Results: Currently, 70 pts were enrolled, of which 24.3% were male. Median age was 68 years, and 35.7% were former/current smokers. Pts with stage IA were 37.1%, IB 24.3%, IIA 2.9%, IIB 11.4%, IIIA 14.3%, IIIB 5.7%. All pts had an EGFR mut, 54.2% detected by NGS on surgical or pre-surgical specimens. EGFR mut of the other samples were detected using PCR Cobas, and NGS are ongoing. EGFR mut was not detected on plasma after surgery (except for 1). Most pts harboured common mut (47.1% ex19del and 45.7% exon21 L858R on COBAS, 65.6% ex19del and 34.4% exon21 L858R on ONCOMINE). Interestingly, of the 43 pts with post-surgery NGS on plasma, 41.9% had a pathogenic mutation ( TP53 50%, ARID1 and BRCA1/2 11.1% each, 5.6% each SMAD4, MPL, TSC1, NOTCH1, JAK2, APC, FGFR1 and KRAS ) with a median VAF of 0.25% (tissue confirmation is needed to exclude hematopoietic origin). Liquid biopsies were obtained from all pts, with at least one sample collected post-surgery at varying time points. Adjuvant/neoadjuvant chemotherapy was administered to 31.4% of pts, while 17.1% received adjuvant osimertinib (100% after adjuvant osimertinib approbation by local label if indicated) and 11.4% also received radiotherapy. Disease recurrence occurred in 22 patients (31.4%), with 8 being local recurrence. At data lock 83% of pts were alive. Conclusions: Evaluation of EGFR in early-stage NSCLC should be standard procedure. We aim to identify a pattern associated with higher risk of recurrence and worse prognosis. Dynamic monitoring of EGFR could aid in personalising adjuvant treatments and follow-up scheduling.