Life

Abstract Background: Acute Lymphoblastic Leukemia (ALL) is a highly heterogeneous pediatric malignancy. Despite high survival rates, relapse and the involvement of central nervous system (CNS) remains a significant clinical challenge. Traditional clinical parameters often lack the precision required for early detection and risk stratification. This study utilizes high-throughput proteomics and machine learning to identify molecular signatures in cerebrospinal fluid (CSF) that characterize disease effect and treatment response. Methods: 82 CSF samples from 41 pediatric ALL patients at diagnosis (VD) and remission (VR) were analyzed. Proteomic profiling of 276 proteins was performed using Olink Proximity Extension Assay. Differentially abundant proteins were identified (q-value< 0.05, |Log_2FC| > 0.5) using the Wilcoxon rank-sum test. Three machine-learning algorithms - Random Forest, LASSO, and SVM-RFE - were integrated to select the differentially abundant proteins in VR and VD and between CNS involvement levels. To validate the data Pan-Cancer Atlas analysis was done using two different platforms. Results: In the remission phase, we observed significant alterations in the expression of key proteins compared to diagnosis, with ADGRG1 and KYNU showing a marked increase, while CCL17, CD5, CD27, CXCL9, CXCL11, FASLG, GZMA, and TNFRSF9 were significantly downregulated. Furthermore, our analysis identified distinct protein signatures associated with CNS involvement: CCL4, CTSC, CXCL10, CXCL9, and MMP7 were differentially abundant at the VD stage, whereas CAIX, CASP-8, HAGH, CXCL9, MMP7, MCP-2, and VWC2 at the VR stage. Conclusion: Integrating Olink proteomics with machine learning identified molecular signatures in ALL that have the potential to be further developed to a biomarker panel for monitoring treatment response and guiding personalized therapeutic strategies shifting the focus toward the Precision One Health approaches.

BackgroundThe physical properties of leukocytes, such as cell size and stiffness, are critical for their circulation in microcapillary networks where rapid shape changes are required to squeeze through vascular constrictions. Alterations in the cells physical phenotype can promote venous thromboembolism (VTE), a common cause of death in cancer patients receiving chemotherapy. While biochemical VTE predictors are well studied, physical properties of blood cells receive less attention. MethodsUsing real-time deformability cytometry (RT-DC), we monitored for the first time the physical phenotype of leukocytes in a longitudinal study of a breast cancer patient treated with epirubicin/cyclophosphamide (EC) and paclitaxel (Pax). ResultsThe leukocyte counts extracted from RT-DC were in good agreement with standard clinical leukograms and EC had no immediate effect on leukocyte properties. However, Pax caused a significant softening of granulo/monocytes and a stiffening of lymphocytes immediately after administration. Leukocyte size was constant throughout the therapy, but we observed an overall increase in leukocyte stiffness, which was restored to normal values 45 weeks post treatment. ConclusionTaken together, our data reveal chemotherapy-induced specific alterations of leukocyte stiffness potentially critical for microcirculation. Thus, RT-DC measurements can add important, yet currently not available information to VTE prediction in cancer patients.

O_LIThe effect of some bioactive compounds on living organisms is dependent on their concentration and gradients, as is the case of hormones and signalling peptides, determining cell identity, activity and organism development. C_LIO_LIThere are a handful of methods that allow to produce spatially confined peaks of concentration local application of biochemicals on plants, such as agar blocks and microinjection, but they lack in precision, throughput and/or simplicity. C_LIO_LIWe developed the MicroTron, a microfluidics-based method specifically for filamentous organisms or life cycle stages, like the moss plant Physcomitrium patens protonemata, that serves as a platform for the application of chemicals on single cells and study the cell response. C_LIO_LIWe show how chemical applications could be performed on cells, either on the side or apically with dyes and hormones, targeting the cell wall, cell membrane, cytosol and nucleus. C_LIO_LITreatments could be applied on single filaments and with a precision of up to single cells in optimal conditions. C_LIO_LIThis method could be used to study live responses to chemicals with high spatiotemporal resolution. C_LI

Emerin (EMD), an inner nuclear membrane protein essential for nuclear architecture integrity, gene expression, cellular signaling, and chromatin stability, interacts with the LINC complex and participates in cytoskeleton-nucleoskeleton communication by binding to nuclear actin filaments. EMD is implicated in migration, invasion, and metastasis in some tumors, but its role in glioblastoma (GBM) remains unclear. This study evaluated the effects of EMD knockdown and overexpression in GBM cell lines following genotoxic treatment with cisplatin. In both wild-type p53 (U87-MG) and mutant p53 (U138-MG) GBM cells, EMD expression is high, and cisplatin treatment did not affect these protein levels. EMD knockdown in U87-MG cells significantly increased cisplatin IC50, viability, and proliferation. Conversely, stable overexpression of EMD in U87-MG cells led to reduced cisplatin IC50, viability, proliferation, and migration. EMD knockdown or overexpression did not affect any U138-MG phenotypes, with or without cisplatin treatment. Modulation of EMD levels causes morphological changes in stress fiber cytoskeleton, whereas overexpression of EMD in U87-MG cells promotes an increase and a decrease in nuclear and cytoplasmic actin levels, respectively. These biological responses of U87-MG cells overexpressing EMD were coincidentally associated with alterations in the levels of pH2AX(Ser139), p-p53(Ser15), p53, and p21Kip1 proteins after cisplatin exposure. In sum, modulation of EMD levels affects the viability, migration, and proliferation of wild-type p53 GBM cells treated with cisplatin, suggesting unknown roles in the DNA damage response and repair. This work highlights EMD as a potential regulator of GBM chemoresistance and a target for therapeutic intervention.

Rationale. Racial disparities in outcomes among patients with acute respiratory failure are well-described, but the contributions of clinicians to these disparities have not been evaluated. Objectives. Among mechanically ventilated patients, we evaluated racial disparities in severity of illness trajectories and adapted value-added modeling to quantify nurse and physician relationships with these disparities. Methods. In a retrospective cohort of mechanically ventilated patients across five hospitals between 2018 and 2022, we used generalized estimating equations to model the change in Laboratory-based Acute Physiology Score version 2 (LAPS) from the start to end of intensive care unit admission ({Delta}LAPS). Consistent with value-added modeling, we randomly allocated the cohort into development and testing partitions, and fit separate multiple linear regression models of {Delta}LAPS using concurrent nurse and physician assignments (determined at 4-hour intervals), patient race, and clinician-race interaction terms as fixed effects. Clinician-specific and clinician-race interaction coefficients were extracted to determine race-specific value-add for each clinician. We defined the race-contextual value-add difference (RCVAD) as a clinician-level measurement of the difference in that clinician's value-add between Black and White patients in their care; a positive RCVAD indicates a more favorable severity of illness trajectory for Black relative to White patients and vice versa. Measurement and Main Results. Among 6,555 distinct patients, 7,247 clinical encounters, 405 nurses, and 70 physicians, Black patients accounted for 2,926 (40%) encounters. Overall, Black patients had significantly less improvement in {Delta}LAPS than White patients (difference in LAPS decline = 2.26 [0.23, 4.29], p=0.029). In the development partition, median nurse RCVAD was -0.10 (interquartile range [IQR]: -1.17, 1.14) with 191 (47%) nurses having a positive RCVAD; median physician RCVAD was -0.18 (IQR: -1.34, 0.56) with 29 (41%) having a positive RCVAD. Conclusions. Black mechanically ventilated patients experience less improvement in severity of illness during intensive care unit admission than White patients. While the majority of physicians and nurses were associated with disparities-exacerbating illness trajectories, many other clinicians were associated with disparities-mitigating trajectories. Future work to understand practices associated with disparities-exacerbating and disparities-mitigating care profiles could inform interventions to reduce disparities overall.

Background: Large language models (LLMs) struggle with dynamic, longitudinal clinical reasoning. We developed a Multi-Stage Iterative Clinical Reasoning Agent framework to address this gap and systematically decouple the clinical efficacy of static retrieval-augmented generation (RAG) from dynamic self-refinement. Methods: Ten complex longitudinal nephrology cases, rigorously selected via a modified Delphi consensus technique, were blindly evaluated by four board-certified nephrologists and a multi-model AI panel. We compared three architectures across nine cognitive steps: (Model A) a baseline frontier LLM, (Model B) an LLM augmented with static guideline-based RAG, and (Model C) our proposed multi-agent framework featuring RAG integrated with iterative self-critique and refinement. Results: In human evaluations (20-point scale), Model C (mean 17.2, SD 1.2) significantly outperformed both Model A (16.1, 1.3) and Model B (16.2, 1.2) (P < 0.001). Implementing static RAG (Model B) yielded no significant improvement over the baseline. Automated AI evaluations (15-point scale) corroborated these findings: Model C (14.7, 0.6) outscored Model A (14.2, 0.9, P < 0.001) and Model B (14.3, 0.9, P = 0.01). While monolithic models exhibited severe score degradations in planning-heavy tasks such as dynamic differential diagnoses, the multi-agent framework effectively intercepted error cascades, achieving significantly higher diagnostic accuracy (mean 17.6, P = 0.019) and therapeutic management scores (17.3, P = 0.002). Conclusions: Static knowledge retrieval alone fails to enhance frontier LLM performance in longitudinal medical reasoning. Distributing clinical workflows into a multi-agent dynamic refinement pipeline significantly improves reasoning completeness, intercepts error cascades, and safely resolves planning bottlenecks in complex patient care.

Abstract Objective: To synthesize the available evidence on the association between SARS-CoV-2-related microvascular thrombosis and acute kidney injury (AKI), with emphasis on renal outcomes, mortality, and renal replacement therapy requirements. Methods: This systematic review followed the PRISMA 2020 statement and was prospectively registered in PROSPERO (CRD420251132701). PubMed/MEDLINE, Scopus, and Embase were searched for systematic reviews, including meta-analyses, and umbrella reviews investigating the association between SARS-CoV-2-related microvascular thrombosis and acute kidney injury. Two reviewers independently performed study selection, data extraction, and methodological quality assessment using AMSTAR-2 and ROBIS. Evidence was synthesized through a structured narrative synthesis supported by quantitative data extracted from the included reviews. Results: Six evidence syntheses evaluating kidney involvement, thrombotic events, and microvascular mechanisms in COVID-19 were included. AKI incidence was 9.2% (95%CI 4.6-13.9) among hospitalized patients and 32.6% (95%CI 8.5-56.6) among critically ill patients. In children with multisystem inflammatory syndrome associated with SARS-CoV-2, AKI incidence was 20% (95%CI 14-28). Microvascular or thrombotic events were associated with adverse renal outcomes (OR 2.14; 95%CI 1.32-3.48). AKI was associated with increased mortality (OR 4.68; 95%CI 1.06-20.70) and greater likelihood of renal replacement therapy requirement (OR 2.87; 95%CI 1.45-5.68). The certainty of evidence ranged from moderate to high for the principal outcomes. Conclusion: Current evidence supports an important association between microvascular thrombotic injury and COVID-19-associated AKI. These findings reinforce the relevance of endothelial dysfunction and thromboinflammatory pathways in kidney involvement during COVID-19 and highlight the need for early renal monitoring, risk stratification, and kidney-protective strategies in high-risk patients. Keywords: COVID-19; Acute Kidney Injury; Microvascular Thrombosis; SARS-CoV-2; Renal Replacement Therapy; Systematic Review

Background: Deceased-donor kidney transplantation is a component of the Brazilian transplant system and takes place within a deceased-donation environment that in-cludes donor identification, notification, family approach and authorization, organ allocation, and logistics. This study described temporal, macroregional, and federative unit-level variation in deceased-donor kidney transplantation in Brazil from 2014 to 2024, together with hospitalization-based indicators and contextual indicators of the deceased-donation environment. Methods: This nationwide descriptive time-series study used publicly available secondary data from the Hospital Information System of the Unified Health System (SIH/SUS), the Brazilian Transplant Registry (RBT/ABTO), and the Brazilian National Transplant System (SNT). Indicators includ-ed the number of transplants, transplant rates per million population, kidney transplant waiting list stock, mean length of hospital stay, in-hospital mortality, potential donor notifications, and family interview and refusal proportions. The three databases were analyzed separately and in parallel, without linkage. Results: The annual number of deceased-donor kidney transplants increased over the study period. The kidney trans-plant waiting list stock also increased. Mean length of hospital stay decreased, and in-hospital mortality decreased over time. Marked macroregional and federative unit-level heterogeneity was observed in transplant activity, hospitalization-based indica-tors, and contextual indicators of the deceased-donation environment. Conclusions: Deceased-donor kidney transplantation increased in Brazil between 2014 and 2024, although regional disparities persisted. These findings support monitoring strategies that incorporate contextual indicators alongside measures of transplant activity. Be-cause this was a descriptive study based on secondary data from distinct sources, the findings should not be interpreted as evidence of causal relationships.

Background: Uremic pruritus affects up to 90% of individuals and substantially impairs quality of life, in the group of patients undergoing chronic dialysis. Despite multiple therapeutic options, an optimal and well tolerated treatment remains elusive. Ketotifen, a mast cell stabilizer with antihistaminic properties prevent itch by inhibition of mast cell derived tryptase, which modulates protease-activated receptor-2 (PAR-2) in the cowhage itch pathway. Materials and Methods: In this prospective observational study, 230 chronic dialysis patients were screened, of whom 48 (20.9%) had clinically significant pruritus identified using a structured questionnaire. Twenty-four patients with moderate-to-severe symptoms who were prescribed ketotifen as part of routine clinical care consented to prospective follow-up. Ketotifen was initiated at 1 mg twice daily, with dose escalation to 2 mg twice daily in patients with persistent symptoms according to routine clinical practice. Pruritus severity was assessed using visual (VAS), verbal (VRS), and numerical (NRS) rating scales before and after treatment. Results: After two weeks of initial 1mg therapy, 19 showed significant clinical improvement. Mean scores reduced 77.5 [-&gt;] 27.1 (VAS), 87.5 [-&gt;] 20.8 (VRS), and 74.2 [-&gt;] 25 (NRS) (around 65% reduction with p < 0.001 across all scales). Clinical relief was achieved in 83.3% overall and mild tolerable drowsiness occurred only at the 2mg dose. Conclusion: Ketotifen is a safe, effective and well tolerated option for moderate to severe uremic pruritus in dialysis patients. Larger multicenter studies are warranted to confirm efficacy and optimize dosing.

Introduction Hospitalised community-acquired pneumonia (CAP) is heterogeneous in aetiology, severity, and outcome. Phenotyping and endotyping approaches offer potential to stratify patients biologically and guide targeted therapy, but require well-characterised cohorts with linked biosamples. We describe the PARIS (Pneumonia: Acute Respiratory Infection +/- Sepsis) study: a prospective observational cohort of hospitalised patients with pneumonia, designed to characterise functional outcomes and to provide a biobank for translational immunological research. Methods Adults admitted with CAP to a single NHS district general hospital were enrolled within 24 hours of admission between December 2020 and March 2022. Clinical, functional, and physiological data were collected at enrolment, hospital discharge, and 6-8 week follow-up. Serial blood samples were collected for flow cytometry, transcriptomics, pathogen DNA detection, and plasma biobanking. Results Forty-seven patients were enrolled (15 without and 32 with sepsis [SOFA >=2] at enrolment); 87% met sepsis criteria by 24 hours post enrolment. Most patients (30/47, 64%) were managed as COVID-19, microbiologically confirmed in 27. Mean age was 57 years (SD 16), 70% were male, and baseline comorbidity burden was low. Severity was moderate (median NEWS2 4 at enrolment, rising to 6 by 24 hours post enrolment; p<0.001). Mortality was 4/47 (8.5%), with 44/47 (94%) alive at hospital discharge. Median length of stay was 8 days (IQR 5.5-11). Translational samples were collected from the majority: fresh flow cytometry (44/47, 94%), transcriptomics from the sepsis subgroup (31/32, 97%), pathogen DNA sampling (35 samples received across study timepoints; see Table 5), and stored plasma (29/47, 62%). The primary outcome of functional decline (Barthel score decrease >=1.85) occurred in only 1/29 patients with paired assessments (3.4%). Persistent CRP elevation (>3 mg/L) at 6-8 week follow-up was present in 16/31 (52%) survivors with available data. Conclusions The PARIS cohort provides a well-characterised clinical platform and linked biobank to support translational studies of pneumonia and sepsis. The low rate of functional decline reflects the younger, lower-comorbidity, COVID-predominant population recruited. Primary protocol endpoints were not achieved owing to pandemic-related disruption. Data and samples underpin a programme of linked translational studies.

Background Hypercapnia may indicate a primary ventilatory syndrome, a complication of another illness, or an epiphenomenon of severe disease. The presenting context of hypercapnia is poorly quantified, limiting clinical interpretation and synthesis of epidemiologic studies. Methods We performed a retrospective cross-sectional study of Medical Information Mart for Intensive Care IV (MIMIC-IV) hospital admissions linked to an emergency department (ED) presentation from 2011 through 2019. Admissions were included if the triage chief complaint was not missing and at least one prespecified criterion for hypercapnia was met: an International Classification of Diseases (ICD) code for hypercapnic respiratory failure or obesity hypoventilation syndrome, arterial blood gas (ABG) PCO2 45 mmHg, venous blood gas (VBG) PCO2 50 mmHg, or indeterminate-source blood gas PCO2 50 mmHg. Triage chief-complaint text was classified by natural language processing (NLP) into 17 National Hospital Ambulatory Medical Care Survey reason-for-visit (RFV) categories using a multi-label framework. Primary analyses estimated admission-level RFV category prevalences; secondary analyses compared distributions by overlapping ascertainment indicator, age, and acidemia. Results The total cohort included 11,941 admissions: 1,542 (12.9%) met both blood-gas and ICD-code criteria, 9,958 (83.4%) met blood-gas criteria only, and 441 (3.7%) met ICD-code criteria only. Median age at admission was 68 years (IQR 56-78), and 6,423 admissions (53.8%) were for male patients. Respiratory RFV categories were most prevalent (30.2%), followed by administrative reasons (17.5%), digestive symptoms (14.0%), injuries and adverse effects (14.0%), and nervous-system symptoms (13.8%); categories were not mutually exclusive. Respiratory categories were more common in ICD-positive admissions (50.2%) than in VBG-defined (36.3%) or ABG-defined admissions (27.3%). Injuries and adverse effects were most prevalent among admissions for patients aged 18-39 years (34.4%), whereas respiratory categories increased from 13.7% among admissions for patients aged 18-39 years to 36.5% among admissions for patients aged 80 years. NLP-derived classifications showed mean set-F1 of 0.84 against adjudicated clinician labels in the full annotated benchmark sample. Conclusions Among ED-linked admissions with hypercapnia by diagnosis code, blood gas, or both, respiratory complaints were the most common chief-complaint category but represented fewer than one-third of admissions. Presentation context should be incorporated when defining, comparing, and interpreting hypercapnia cohorts, particularly those ascertained by blood-gas criteria.

Colorectal cancer (CRC) is challenging to track because its molecular changes are very complex as the disease progresses, creating significant challenges for robust biomarker discovery. In this study, we developed a machine learning framework by integrating monotonic progression and the StepMiner approach. We conducted external validation to identify reproducible, consistent transcriptomic biomarkers associated with CRC progression. Gene expression datasets were analyzed across four disease states from publicly available GEO: normal colon, adenoma, primary colorectal cancer, and metastasis. First, we identified genes with monotonic expression, then used the StepMiner approach to identify genes that act as switches between stages. A balanced 74-gene signature was used for machine-learning classification with a Random Forest. External validation showed strong performance in tissue-based datasets. However, tissue-derived signatures and plasma and blood-based datasets showed poor performance, highlighting biological differences between transcriptomic profiles. Cross-filtering between tissue-derived genes and blood expression datasets was performed, which resulted in the selection of 62 blood-compatible gene signatures. Leakage-free retraining on GSE164191 achieved a mean AUC of 0.868 with balanced precision. Functional enrichment analysis showed that these genes are highly active in cancer growth. Specifically, genes CBX3, S100A11, PDK4, NCOR1, and SOX4 demonstrated stable and reliable performance across the validation fold. Overall, our study presents a progression-aware transcriptomic framework for CRC biomarker discovery and demonstrates the importance of external validation. Additionally, we evaluate whether tissue-derived signatures can predict blood profiles. This proposed approach may help the future development of tissue-based diagnostics and minimally liquid-biopsy strategies for CRC. To ensure reproducibility, our proposed workflow was automated as a Nextflow pipeline. The tissue-derived model was deployed as an application utilizing Angular, ASP.NET Core, and Plumber (R).

Abstract Familial kidney disease is common in Cyprus. Patients with a glomerular phenotype are most likely to have an autosomal dominant variant in collagen type IV alpha 3 chain (COL4A3) or collagen type IV alpha 4 chain (COL4A4) genes but pathogenic variants are not found in the majority of families. We compare the clinical phenotype between two groups of 10 Turkish Cypriot families who lack a pathogenic variant of COL4A3 or COL4A4 but have either the COL4A4 variant p.G545A or p.G999E. Both groups had identical clinical phenotypes with microscopic haematuria detected at least once in 76% of affected family members; urine protein was less than 1 g/day until glomerular filtration rate (GFR) was <30 ml/min. End-stage kidney disease (ESKD) occurred in 24.1% of those over 50 at a median age of 62.8 (36-86) years. Although the genetic cause of renal injury in this large group is still unknown, these families present with a clinical phenotype best characterised as familial hypertensive nephropathy. We propose that this condition accounts for the great excess of renal failure in the Eastern Mediterranean in those over 65 years of age.

Rising costs, slow accrual and molecular substratification of cancers necessitate novel clinical trial designs. We demonstrate that artificial intelligence-generated synthetic patients can replace real controls to reproduce results of the SORAML trial. Using external multimodal data from 1,377 acute myeloid leukemia (AML) patients from previous trials and a real-world registry, we fine-tuned a tabular foundation model to generate synthetic patients, reproducing clinical and genetic features and outcome associations. Synthetic patients were then matched to the original SORAML intervention group using Cox risk scores, replacing the original control and reproducing the original trial result with near-identical median event-free survival (EFS) and treatment effect (original hazard ratio [HR] 0.64, 95%-confidence interval [CI] 0.47-0.87, p=0.004; with synthetic control HR 0.66, 95%-CI 0.48-0.90, p=0.009). Our findings demonstrate that AI-generated synthetic patients can serve as statistically rigorous controls supporting novel trial designs.

Anoxia is a major stress for most vertebrates and frequently accompanies harsh winter conditions, particularly in species that spend much of the season frozen solid. North American freeze-tolerant wood frogs (Rana sylvatica) can survive several months without oxygen and endure whole-body freezing for up to eight months of the year, with [~]70% of total body water frozen as extracellular ice, yet revive when temperatures rise in spring. Survival depends on multiple adaptations, including tolerance of prolonged oxygen deprivation while frozen, when breathing and circulation are halted. A key strategy involves hepatic glycogen mobilization, producing large amounts of glucose that are distributed to tissues where it functions both as a cryoprotectant and as a substrate for anaerobic ATP production. The present study examines the role of histone lysine methylation and demethylation in regulating liver proteins under anoxic conditions. Relative protein expression of seven histone methyltransferases (ASH2L-S, ASH2L-L, RBBP5, SETD8, SMYD2, ESET, SETD1), six lysine demethylases (KDM1A, KDM3B, KDM4A, KDM4B, KDM5A, KDM5C), and eight histone marks (H3K4me1, H3K4me2, H3K9me3, H3K27me3, H3K36me3, H3K79me3, H4K20me1, H4K20me3) were evaluated in wood frog liver under control, 4-hour, and 24-hour anoxia exposures. The data indicate that histone lysine methylation and demethylation contribute significantly to transcriptional regulation under anoxia. Specifically, H3K4, H3K36, and H3K79 methylation were associated with transcriptional activation, whereas H3K9, H3K27, and H4K20 methylation correlated with transcriptional repression. These findings highlight the dynamic role of epigenetic regulation in supporting hypometabolism and stress adaptation in freeze-tolerant wood frogs.

Chronic hepatitis B virus (HBV) infection contributes to hepatocellular carcinoma by disrupting host transcription, cell-cycle control, and apoptotic signaling. Isochlorogenic acid A (ICAA), a natural compound with antiviral and hepatoprotective properties, was previously shown to inhibit HBV replication by interfering with multiple steps of the viral life cycle. Because chronic HBV often reflects an imbalance between proliferation and cell death, we investigated how ICAA affects gene expression related to these processes in the presence or absence of HBV. We performed transcriptome analysis using RNA sequencing (RNA-seq) in HepAD38 cells (a HepG2-derived stable HBV-expressing line) and HepG2 control cells (HBV-negative) treated with ICAA or DMSO. HBV caused major differences in gene expression in HepAD38 cells compared with HBV-negative HepG2 cells. Principal component analysis showed that ICAA significantly altered HBV-dependent expression patterns, resulting in 189 differentially expressed genes (DEGs) that were regulated in opposite directions by both HBV and ICAA. Functional enrichment analysis highlighted pathways in viral carcinogenesis, apoptosis, MAPK signaling, and p53 signaling. Annexin V/propidium iodide assays showed apoptotic cells in both treated and untreated HepAD38 cultures, with only minor pattern changes. Mechanistically, in untreated HBV-positive cells caspase-9 cleavage failed to activate PARP, suggesting that induction of intrinsic apoptosis is followed by blocked execution. In contrast, ICAA inhibits caspase-9 cleavage in a dose-dependent manner, while activating PARP. Consistent with this, ICAA treatment increased apoptotic DNA fragmentation in HepAD38, reflecting the proapoptotic potential of ICAA under these conditions facilitating the elimination of HBV-positive cells by apoptosis. These findings highlight the potential therapeutic relevance of this compound in processes associated with HBV pathogenesis, together with its antiviral effect. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=185 SRC="FIGDIR/small/733975v1_ufig1.gif" ALT="Figure 1"> View larger version (47K): org.highwire.dtl.DTLVardef@38d107org.highwire.dtl.DTLVardef@235a13org.highwire.dtl.DTLVardef@ee988aorg.highwire.dtl.DTLVardef@60cb13_HPS_FORMAT_FIGEXP M_FIG C_FIG

Background: Endothelial progenitor cells (EPCs) contribute to endothelial repair and neovascularization, and EPC dysfunction is closely associated with oxidative stress-related vascular injury. Forkhead box O3a (FoxO3a) regulates cellular stress responses, whereas miR-34a has been implicated in endothelial dysfunction, senescence, and apoptosis. However, the relationship between FoxO3a and miR-34a-3p in oxidatively injured EPCs remains incompletely defined. Objective: This study investigated the role of FoxO3a in H2O2-induced EPC dysfunction and examined whether miR-34a-3p directly interacts with the FoxO3a 3' untranslated region (3'UTR). Methods: Human umbilical cord blood-derived EPCs were identified by DiI-ac-LDL uptake, FITC-UEA-1 binding, and the expression of EPC-related markers. Oxidative stress was induced by H2O2. Cell viability, apoptosis, and angiogenic capacity were evaluated using CCK-8 assay, Annexin V/7-AAD flow cytometry, and Matrigel tube formation assay, respectively. FoxO3a expression was modulated using adenoviral overexpression or knockdown vectors, and miR-34a was modulated using mimics or antagomir. FoxO3a and miR-34a expression levels were detected by Western blot and qPCR. A dual-luciferase reporter assay was used to verify the interaction between hsa-miR-34a-3p and the FoxO3a 3'UTR. Results: H2O2 reduced EPC viability, increased apoptosis, and impaired tube formation in a concentration-dependent manner. H2O2 increased FoxO3a protein abundance and miR-34a expression, whereas FoxO3a mRNA did not change markedly. FoxO3a overexpression aggravated, whereas FoxO3a knockdown partially alleviated, H2O2-induced EPC dysfunction. Similarly, miR-34a mimics further suppressed EPC viability and tube formation, while miR-34a antagomir exerted a protective effect. Dual-luciferase reporter analysis showed that hsa-miR-34a-3p significantly reduced the activity of the wild-type FoxO3a 3'UTR reporter, while mutation of the predicted binding site abolished this suppression. Conclusion: FoxO3a and miR-34a participate in oxidative stress-induced EPC dysfunction. The dual-luciferase data demonstrate that hsa-miR-34a-3p directly targets the FoxO3a 3'UTR, suggesting the presence of miR-34a-3p-mediated post-transcriptional feedback within the FoxO3a-related stress-response network in EPCs.

Introduction People with kidney failure receiving haemodialysis experience significantly high rates of depression. However, there is a lack of evidence on how to treat depression in this population. A significant barrier to effective treatment is the high treatment burden of haemodialysis, which makes additional appointments prohibitive. Behavioural activation (BA) is an evidence-based brief therapy for depression that has been delivered in a variety of different clinical settings. However, it has not previously been evaluated in the haemodialysis setting. Methods This study aims to evaluate the feasibility and acceptability of a cluster randomised controlled trial (cRCT) of intradialytic BA for people with kidney failure. The study consists of three main components: A pilot cRCT, where we will recruit 52 people who are receiving haemodialysis and experiencing symptoms of depression across two sites. Patients will be cluster-randomised to either BA or usual care and will be followed up for three months; a qualitative process evaluation using semi-structured interviews to explore the experiences of patients, healthcare professionals and carers; and a feasibility economic evaluation exploring the feasibility of collecting healthcare resource use data. Results Key findings will include the feasibility of screening and recruiting participants, participant retention, completion of clinical outcome measures, and the acceptability of the intervention. Conclusion If feasible, the next step will be to conduct a definitive, adequately powered cRCT to determine the effectiveness of the intervention in this population, so that we can improve the identification and management of depression for people with kidney failure.

Biomolecular condensates are widespread cellular self-assembled structures with essential functions. There are suggestions of condensates formed by different proteins being near criticality. However, systematic investigation of the criticality of condensates is absent, and critical exponents defining their universality class have not been found. Here, using long-time simulations, we show that condensates exhibit typical critical phenomena, including scale-free spatiotemporal correlations, critical slowing down, divergence of correlation length and dynamic scaling. From these scaling behaviors, a set of critical exponents is determined. Based on dynamic critical exponent, diverse condensates can be divided into two distinct universality classes, arising from differences in their molecular components and interaction types.

Predicting the long-term kidney function decline is critical for timely intervention but remains challenging. While the urinary protein-to-creatinine ratio (uPCR) is a potential surrogate endpoint, its short-term reduction's link to long-term nephroprotection requires investigation. This study aimed to develop a probabilistic neural network model to capture both the estimated glomerular filtration rate (eGFR) slope and its uncertainty based on baseline clinical characteristics. Using a retrospective dataset, we designed a neural network to output a predictive distribution (mean and standard deviation {sigma}) for the eGFR slope. SHAP (SHapley Additive exPlanations) was used for model interpretation, and a simulation study quantified the impact of uPCR reduction. In the validation set, the model achieved a Pearson's correlation coefficient of 0.56 and an RMSE of 2.81 ml/min/1.73m^2/year between predicted and actual slopes. SHAP analysis identified uPCR as the most potent predictor, with higher baseline levels associated with a more rapid eGFR decline. Furthermore, a simulated 62% uPCR reduction demonstrated a significant improvement in the predicted eGFR slope, an effect most pronounced in patients with high baseline uPCR. This proof-of-concept study reinforces the critical role of uPCR in predicting eGFR slope and suggests its reduction may contribute to long-term kidney function preservation, warranting validation in larger, diverse real-world datasets.