Fibronectin type III domain-containing protein 4 (FNDC4) is a circulating hepatokine with anti-inflammatory and cardioprotective properties, but its prognostic relevance after ST-segment elevation myocardial infarction (STEMI) remains uncertain. We investigated whether admission serum FNDC4 levels were associated with 12-month major adverse cardiovascular events (MACE) in patients with STEMI undergoing primary or urgent percutaneous coronary intervention (PCI). This single-center prospective observational study included 274 patients. Serum FNDC4 was measured before PCI using an enzyme-linked immunosorbent assay, and participants were followed for 12 months. Associations and prognostic performance were assessed using Spearman correlation, receiver operating characteristic, Kaplan–Meier, and Cox proportional hazards analyses. During follow-up, 51 patients (18.6%) experienced MACE. Serum FNDC4 levels were significantly lower in patients with MACE and were inversely correlated with interleukin-6 (IL-6) and N-terminal pro–B-type natriuretic peptide (NT-proBNP). The area under the receiver operating characteristic curve (AUC) for FNDC4 was 0.771 (95% confidence interval [CI], 0.695–0.839). Adding FNDC4 to a clinical model increased the AUC from 0.874 to 0.905 (ΔAUC, 0.031; 95% CI, 0.009–0.061). After adjustment for age, NT-proBNP, left ventricular ejection fraction (LVEF), estimated glomerular filtration rate (eGFR), and Killip class III/IV, each 10 ng/mL decrease in FNDC4 was associated with a higher risk of MACE (hazard ratio, 1.821; 95% CI, 1.360–2.438; p<0.001). Lower admission FNDC4 levels were independently associated with 12-month MACE after PCI for STEMI and may provide complementary prognostic information; however, multicenter external validation and serial FNDC4 measurements are required before clinical application.
The widespread adoption of contemporary human epidermal growth factor receptor 2 (HER2)-targeted therapies has altered survival in HER2-positive metastatic breast cancer, potentially limiting the applicability of older prognostic models. We aimed to develop and internally validate nomograms for overall survival (OS) and breast cancer-specific survival (BCSS) in patients with de novo stage IV HER2-positive breast cancer diagnosed during 2018–2021. Using the Surveillance, Epidemiology, and End Results (SEER) database, 1,516 patients were randomly assigned to training (n = 1,061) and internal hold-out validation (n = 455) cohorts. Random Survival Forest analysis with a 98% out-of-bag concordance index criterion selected variables for multivariable Cox regression models predicting 1-, 3-, and 5-year survival. Discrimination, calibration, clinical utility, and risk stratification were assessed. Nine variables were retained for OS and seven for BCSS. Concordance indices in the training and validation cohorts were 0.764 and 0.741 for OS and 0.747 and 0.736 for BCSS, respectively. Areas under the time-dependent receiver operating characteristic curves (AUCs) ranged from 0.73 to 0.82. Calibration was generally acceptable, and decision curve analysis suggested potential net clinical benefit. High-risk patients had markedly poorer OS (hazard ratios, 5.05 in training and 5.07 in validation) and BCSS (4.32 and 4.09, respectively; all p < 0.001). Sensitivity analyses addressing the time-varying chemotherapy effect and competing risks broadly supported the primary findings. The nomograms showed good internal discrimination and may support individualized prognostic assessment once initial treatment information is available. However, SEER does not record specific HER2-targeted regimens; therefore, this cohort represents a contemporary calendar period rather than a treatment-confirmed dual-HER2-blockade population. External validation in treatment-annotated cohorts is required, and 5-year estimates should be interpreted cautiously given the median follow-up of 27 months.
Anesthesia and surgical stress may produce distinct neuronal and metabolic responses, but comparative data on perioperative biomarker trajectories after general and spinal anesthesia remain limited. This prospective observational study aimed to compare circulating neuron-specific enolase (NSE), adiponectin, and visinin-like protein-1 (VSNL1) in adults undergoing elective surgery under general anesthesia (GA) or spinal anesthesia (SA). Ninety-seven patients (GA, n = 52; SA, n = 45) underwent serum sampling preoperatively and at 6 and 24 hours postoperatively, and biomarker concentrations were measured using enzyme-linked immunosorbent assay (ELISA). Longitudinal associations were assessed using linear mixed-effects models adjusted for age, body mass index (BMI), operation duration, and surgical procedure type, with additional baseline-adjusted analyses for adiponectin. NSE increased in both groups but rose less in the SA group than in the GA group at 6 hours (adjusted interaction estimate, -7.765 ng/mL; p < 0.001) and 24 hours (-4.005 ng/mL; p < 0.001). Adiponectin decreased postoperatively, but group-by-time interactions were not significant, and baseline-adjusted analyses showed no significant association between anesthesia modality and adiponectin at either postoperative time point. VSNL1 increased over time; the group-by-time interaction was not significant at 6 hours but was significant at 24 hours (estimate, 0.635 ng/mL; p < 0.001), indicating a differential late trajectory. Perioperative biomarker trajectories differed between GA and SA, most consistently for NSE. However, nonrandom anesthesia assignment, substantial differences in surgical case mix and operation duration, and the absence of neurocognitive outcome assessment preclude causal or clinical interpretation. These findings should be regarded as exploratory adjusted associations and validated in larger, more homogeneous surgical cohorts.
Human-pathogenic orthohantaviruses are significant rodent-borne zoonotic pathogens in Europe. Southeastern Europe (SEE) is a distinct endemic region where Dobrava-Belgrade virus (DOBV) and Puumala virus (PUUV) co-circulate, causing hemorrhagic fever with renal syndrome (HFRS) of varying severity. This narrative review synthesizes the epidemiology, transmission dynamics, clinical characteristics, ecological drivers, and public health challenges of hantavirus infections across SEE, identifying priorities for regional preparedness. We searched PubMed, Scopus, and Web of Science from their inception through June 2026, supplementing with reference screening. Regional epidemiological patterns were assessed descriptively using national surveillance data, European surveillance reports, published studies, and available population-based incidence estimates. Long-term surveillance reveals persistent endemic transmission with significant geographic and interannual heterogeneity. Slovenia and Croatia exhibit the highest cumulative burden and most pronounced periods of increased HFRS activity. Several years-2008, 2012, 2014, 2017, 2019, 2021, and 2023-showed partially synchronized increases across multiple neighboring countries. These patterns align with shared ecological influences, including rodent population fluctuations, mast-seeding events, and climatic variability. However, local ecology, human exposure, and differences in surveillance capacity modify their magnitude. The coexistence of generally milder PUUV-associated disease and more severe DOBV-associated HFRS further distinguishes SEE's clinical landscape. Therefore, hantavirus epidemiology in SEE is best understood as an interconnected regional ecological system. Integrated human, animal, environmental, and genomic surveillance within a coordinated One Health framework is crucial for improving outbreak prediction, cross-border preparedness, and future disease control.
Recurrent aphthous ulceration (RAU) is a prevalent inflammatory condition affecting the oral mucosa. However, the localized epithelial junctional changes and their connection to immune cell distribution are not well understood. This study sought to characterize junction-associated protein expression and compartment-specific lymphocyte distribution in RAU. This retrospective, multicenter, paired tissue study included 56 consecutive patients with clinically and histopathologically confirmed RAU. Semiquantitative immunohistochemistry was employed to compare E-cadherin, claudin-1, and claudin-6 expression. Comparisons were made between ulcer-adjacent non-ulcerated regenerative epithelium and morphologically preserved distant mucosa from the same patient. The study also evaluated cluster of differentiation 4 (CD4)-, cluster of differentiation 8 (CD8)-, and cluster of differentiation 20 (CD20)-positive lymphocytes in both intraepithelial and subepithelial/ulcer-base compartments. All distant mucosal samples exhibited strong expression of the three junctional proteins. In contrast, within the ulcer-adjacent epithelium, E-cadherin expression was weak in 35.7% and moderate in 64.3% of cases. Claudin-1 was absent in 17.9% and weak in 82.1%, while claudin-6 was absent in 71.4% and weak in 28.6%. Each marker demonstrated significantly lower expression in all paired cases (p < 0.001). Descriptively, CD8-positive lymphocytes were consistently present intraepithelially in all cases, whereas CD20-positive lymphocytes were absent. The ulcer base and adjacent subepithelial tissue contained CD4-, CD8-, and CD20-positive lymphocytes in all cases. RAU is consistently linked to a localized reduction of epithelial junctional proteins and distinct compartmental lymphocyte patterns. These findings suggest epithelial junctional remodeling within the RAU microenvironment, though they do not confirm functional barrier impairment or causality.
In-hospital mortality remains an important concern in patients with ST-elevation myocardial infarction (STEMI), highlighting the need for simple and readily available markers for early risk stratification. This study investigated the prognostic value of the blood urea nitrogen-to-serum albumin ratio (BAR) for in-hospital mortality in patients with STEMI undergoing primary percutaneous coronary intervention (pPCI). This single-center retrospective cohort study included 388 consecutive patients who underwent pPCI in 2024. Admission BAR was calculated using blood urea nitrogen (mg/dL) and serum albumin (g/L). Its association with in-hospital mortality was evaluated using receiver operating characteristic analysis and exploratory multivariable logistic regression, with Firth-penalized regression and bootstrap validation performed as sensitivity analyses. In-hospital death occurred in 18 patients (4.6%). Patients who died had significantly higher BAR values than survivors (0.7 vs. 0.4, p < 0.001). BAR showed good discrimination for in-hospital mortality, with an area under the curve of 0.869 (p < 0.001); the bootstrap-corrected area under the curve was also 0.869. An exploratory cut-off of 0.475 yielded 83.3% sensitivity and 75.4% specificity. In the multivariable model, each 0.1-unit increase in BAR was associated with higher odds of in-hospital mortality (odds ratio 1.412, 95% confidence interval 1.161–1.718; p = 0.0006), with a similar estimate in the Firth-penalized analysis (odds ratio 1.346, 95% confidence interval 1.121–1.616; p = 0.0008). Admission BAR may represent a simple candidate marker for early mortality risk stratification in STEMI patients undergoing pPCI; however, its prognostic performance and clinical utility require validation in larger prospective multicenter cohorts.
Sepsis-induced myocardial dysfunction (SIMD) is a frequent and life-threatening complication of sepsis driven by interconnected inflammatory, oxidative, metabolic, and mitochondrial disturbances. Post-translational modifications (PTMs) have emerged as key regulators of these processes by dynamically altering protein activity, stability, localization, and signaling. This narrative review aimed to summarize the roles of major PTMs in SIMD and evaluate their potential as therapeutic targets. Relevant original studies and authoritative reviews published predominantly within the past decade were identified through targeted searches of PubMed and Web of Science and qualitatively synthesized. Current evidence indicates that phosphorylation, acetylation, ubiquitination, protein methylation, ADP-ribosylation, and palmitoylation regulate major pathways involved in myocardial inflammation, oxidative stress, mitochondrial dysfunction, metabolic reprogramming, apoptosis, and impaired contractility. Crosstalk among these modifications further integrates signaling networks, while upstream epigenetic mechanisms, including DNA methylation and N6-methyladenosine RNA modification, can influence PTM-related enzymes and downstream signaling. Experimental studies also suggest that pharmacological modulation of PTM-regulated pathways may attenuate myocardial injury; however, most mechanistic and therapeutic evidence derives from animal models and cultured cells, with limited validation in human septic myocardium. PTMs therefore represent promising mechanistic and therapeutic targets in SIMD, but systematic characterization of PTM crosstalk and rigorous human translational studies are required before PTM-directed strategies can be incorporated into clinical practice.
Transforming growth factor-β (TGF-β) contributes to fibrosis, immunosuppression, and tumor progression, but systemic TGF-β inhibition is limited by toxicity and lack of selectivity. This exploratory proof-of-concept study evaluated novel peptides designed to target the TGF-β receptor I/II complex and examined their effects on canonical and noncanonical TGF-β signaling in vitro. Peptides 2_5 and 2_6, selected by in silico docking of receptor–ligand interface-derived sequences, were tested in human embryonic kidney 293T (HEK293T) cells at 20 and 50 μM, with the TGF-β receptor I inhibitor SD-208 as a reference. Total and phosphorylated SMAD2, SMAD3, extracellular signal-regulated kinase 1/2 (ERK1/2), and c-Jun N-terminal kinase (JNK) were assessed by Western blotting and densitometry. Peptide 2_5 showed the strongest activity, reducing total ERK1/2 by 65% at 50 μM and phosphorylated ERK1/2 by 90.4% and 83.4% at 20 and 50 μM, respectively; SD-208 reduced total and phosphorylated ERK1/2 by 64.3% and 54.5%, respectively. Peptide 2_5 also reduced total SMAD2 by 81% at 50 μM, whereas its effect on SMAD2 phosphorylation was inconsistent. Peptide 2_6 showed minimal effects, JNK was largely unaffected, and SMAD3 results were inconclusive because of weak signals. These preliminary findings suggest that peptide 2_5 may differentially modulate TGF-β signaling, with a stronger effect on ERK1/2 than on SMAD2; however, receptor targeting and pathway selectivity require confirmation in fully replicated quantitative studies.
Most women with operable breast cancer increasingly survive long enough for non-cancer causes to become important contributors to mortality, particularly at older ages. We investigated whether the recorded sequence of systemic therapy relative to surgery among women receiving chemotherapy was associated with circulatory mortality and examined how causes of death evolve in long-term survivors. This retrospective population-based cohort study included 74,233 women with stage II–III non-metastatic invasive breast cancer diagnosed between 2010 and 2015 in the Surveillance, Epidemiology, and End Results (SEER) database who received chemotherapy and definitive surgery. A six-month landmark was used to reduce immortal-time bias, and treatment groups were balanced using stabilized inverse-probability-of-treatment weighting. Weighted cumulative incidence, cause-specific hazard, and Fine–Gray subdistribution models were used to evaluate circulatory mortality, while mortality patterns after a five-year landmark were examined descriptively by age. Median follow-up was 10.8 years. The weighted 10-year cumulative incidence of circulatory death was 2.06% after recorded preoperative systemic therapy and 2.10% after postoperative therapy, corresponding to a risk difference of −0.05 percentage points (95% confidence interval [CI], −0.31 to 0.18). The cause-specific hazard ratio was 1.092 (95% CI, 0.968–1.231), and the subdistribution hazard ratio was 1.024 (95% CI, 0.920–1.140). Among deaths occurring after the five-year landmark, the circulatory share increased with age, reaching 25.5% in women aged ≥75 years at diagnosis; however, other non-circulatory causes remained the largest category (49.9%). In women aged ≥75 years who survived five years, circulatory mortality approached and eventually exceeded breast-cancer mortality, but the difference was not significant at the prespecified seven-year time point. The recorded systemic-therapy/surgery sequence was not associated with long-term circulatory mortality. In older long-term breast-cancer survivors, late mortality is predominantly multimorbid rather than circulatory-dominant, supporting survivorship care that addresses cardiovascular risk within a broader framework of age-related competing health risks.
Valproic acid (VPA) is widely used to treat neurological and psychiatric disorders but may cause hepatotoxicity partly through oxidative stress and impaired cellular metabolism. This study investigated whether black mulberry (Morus nigra L.) fruit extract attenuates VPA-induced injury in rat FaO hepatoma cells. Cells were exposed to VPA acutely (500 μg/mL for 28 hours) or chronically (120 μg/mL for 4 weeks). Mulberry extract (0.1 mg/mL) was added 3 hours before acute VPA exposure and maintained throughout co-treatment or administered during the final 24 hours of chronic exposure. Cell viability, hydrogen peroxide production, antioxidant enzyme activities, hepatocellular injury markers, glucose concentration, and extracellular 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical-scavenging capacity were assessed. Compared with VPA alone, mulberry extract significantly increased glutathione peroxidase (GPx) activity under both acute and chronic conditions. During chronic VPA exposure, it also increased glutathione S-transferase (GST) activity and extracellular ABTS radical-scavenging capacity, improved cell viability, and reduced extracellular alanine aminotransferase (ALT) activity. During acute exposure, mulberry significantly reduced extracellular aspartate aminotransferase (AST) activity, whereas the improvement in cell viability was not statistically significant. Hydrogen peroxide production did not differ among acute treatment groups, and mulberry did not restore superoxide dismutase or catalase activity. These findings indicate that black mulberry fruit extract partially mitigates VPA-induced hepatocellular injury, primarily by enhancing glutathione-dependent antioxidant defenses. Further dose-response and in vivo studies are required to confirm its hepatoprotective potential.
Pancreatic ductal adenocarcinoma (PDAC) is often diagnosed at an advanced stage, highlighting the need for minimally invasive biomarkers capable of distinguishing it from related risk conditions. This study evaluated relative leukocyte telomere length (LTL) and gut microbiome profiles as potential biomarkers for PDAC. The study included 244 participants with PDAC (n = 37), chronic pancreatitis (CP; n = 56), type 2 diabetes mellitus (T2DM; n = 99), or no related risk conditions (controls; n = 52). LTL was measured by monochrome multiplex quantitative polymerase chain reaction (qPCR), while gut microbiome composition was assessed by 16S ribosomal RNA (rRNA) gene sequencing in a subset of participants with PDAC (n = 12), CP (n = 13), and controls (n = 7). LTL varied with age, sex, and smoking status. After adjustment for these factors, patients with PDAC had significantly longer LTL than controls (p = 0.029), whereas differences between PDAC and CP or T2DM were not significant. LTL showed limited ability to discriminate PDAC from controls (area under the receiver operating characteristic curve = 0.651) and was not associated with overall survival. Gut microbiome diversity and taxonomic composition did not differ significantly among the analyzed groups. These findings indicate that LTL has limited utility as a standalone diagnostic or prognostic biomarker for PDAC but may warrant further evaluation as a complementary marker in larger prospective studies. The exploratory microbiome findings also require validation in adequately powered cohorts.
Rheumatoid arthritis (RA) is a chronic immune-mediated disease in which persistent inflammation and oxidative stress contribute to progressive joint damage. This study evaluated the prophylactic effects of sabinene in complete Freund’s adjuvant (CFA)-induced arthritis and explored potential molecular associations using integrated in vivo and computational approaches. Rats were allocated to vehicle control, arthritic control, piroxicam, or sabinene groups (15, 30, or 60 mg/kg; n = 6/group); sabinene was administered orally beginning 30 minutes before CFA injection and continued for 28 days. Paw swelling, hematological and biochemical parameters, inflammatory and oxidative-stress markers, and gene expression were assessed, followed by network pharmacology, enrichment analysis, and molecular docking. Sabinene dose-dependently attenuated CFA-induced paw swelling and body-weight loss and improved hematological, hepatic, renal, and inflammatory parameters. Treatment also enhanced antioxidant defenses and reduced lipid peroxidation, prostaglandin E₂, 5-lipoxygenase, and anti-cyclic citrullinated peptide antibody levels. Sabinene reduced messenger RNA expression of nuclear factor kappa B (NF-κB), toll-like receptor 4 (TLR4), NOD-like receptor family pyrin domain-containing 3 (NLRP3), caspase-1, gasdermin D, and other pro-inflammatory genes while increasing interleukin-4 and interleukin-10 expression. Network pharmacology identified 133 overlapping sabinene–RA targets, and molecular docking predicted interactions with tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), signal transducer and activator of transcription 3 (STAT3), and interferon gamma (IFN-γ). These findings support a prophylactic protective effect of sabinene against CFA-induced arthritis associated with reduced inflammation, improved antioxidant status, and modulation of inflammasome-related gene expression. Further protein-level, functional, and post-induction studies are required to confirm the proposed mechanisms and therapeutic relevance.
Brain death (BD) is accompanied by systemic inflammation and endothelial dysfunction, but temporal changes in endothelial injury during progression to BD remain insufficiently characterized. This study aimed to evaluate serial changes in the Endothelial Activation and Stress Index (EASIX) during the clinical course leading to BD. This retrospective, single-center study included patients diagnosed with BD in the intensive care unit (ICU) between January 2019 and June 2025. EASIX, calculated from lactate dehydrogenase (LDH), serum creatinine, and platelet count, was assessed at ICU admission, 24 hours before BD, 12 hours before BD, and at BD diagnosis. Temporal differences were analyzed using the Friedman test, followed by Wilcoxon signed-rank tests with Bonferroni adjustment. Of 222 patients diagnosed with BD, 52 met the inclusion criteria. EASIX values differed significantly across the four time points (p < 0.001), with significant differences observed between ICU admission and BD diagnosis and between 24 and 12 hours before BD. These findings indicate that EASIX increases during the clinical course leading to BD and may reflect progressive systemic endothelial activation and worsening critical illness rather than a BD-specific process. Prospective comparative studies are needed to validate its clinical utility.
Human milk oligosaccharides (HMOs) are bioactive components of human milk that support gut microbial development and may also contribute to early brain maturation and cognitive function. This narrative review aimed to synthesize preclinical and clinical evidence on the associations between HMOs, brain structure, and neurodevelopment and to examine the biological mechanisms underlying these effects. PubMed, Scopus, and Web of Science were searched in March 2025 for animal and human studies evaluating specific HMOs in relation to cognitive, behavioral, neurobiological, or neuroimaging outcomes. Of 1,238 records identified, 18 studies met the eligibility criteria and were included in a qualitative thematic synthesis. Preclinical studies indicated that 2′-fucosyllactose (2′-FL) and sialylated HMOs, including 3′-sialyllactose (3′-SL) and 6′-sialyllactose (6′-SL), may improve learning, memory, emotional regulation, and hippocampal long-term potentiation (LTP), although findings varied across experimental models. Proposed mechanisms included vagus nerve-mediated gut–brain signaling, modulation of the gut microbiota, altered neurotransmitter and neurotrophic signaling, and support of synaptogenesis and myelination. Human observational studies associated higher concentrations of 2′-FL, 3′-SL, 6′-SL, and specific HMO combinations during early lactation, particularly around one month postpartum, with better cognitive, language, motor, and executive-function outcomes. Magnetic resonance imaging (MRI) findings further suggested HMO-specific associations with cortical maturation, white-matter connectivity, and myelination. Current evidence supports a potential role for HMOs in early neurodevelopment; however, causal effects remain unconfirmed. Larger, diverse longitudinal studies and randomized controlled trials using standardized neurodevelopmental outcomes are needed to determine optimal HMO compositions, doses, and periods of exposure.
Stenotrophomonas maltophilia is an opportunistic, multidrug-resistant (MDR), Gram-negative bacterium that has emerged as an important cause of hospital- and community-acquired infections, particularly in immunocompromised, critically ill, and hospitalized patients. Its increasing clinical relevance is driven by extensive virulence determinants, biofilm formation, intrinsic and acquired antimicrobial resistance, and persistence within healthcare-associated environmental reservoirs. This narrative review summarizes current evidence on the epidemiology, pathogenic mechanisms, antimicrobial resistance, and therapeutic management of S. maltophilia, with a particular focus on emerging treatment strategies. A literature search of PubMed, Scopus, Web of Science, and Google Scholar was performed to identify relevant studies published between 1997 and 2026. The available evidence demonstrates that biofilm formation, quorum sensing, adhesion factors, secretion of extracellular enzymes, and multiple resistance mechanisms—including β-lactamases, multidrug efflux pumps, and horizontally acquired resistance genes—substantially contribute to persistent infections and therapeutic failure. Hospital outbreaks are frequently associated with contaminated water systems, sinks, and medical devices, emphasizing the importance of environmental surveillance and infection control. Trimethoprim-sulfamethoxazole (TMP-SMX) remains the recommended first-line therapy, whereas minocycline, tigecycline, fluoroquinolones, and cefiderocol represent important alternatives for selected patients. Emerging approaches, including novel tetracyclines, bacteriophages, bacteriocins, ceragenins, anti-biofilm agents, and plant-derived compounds, have demonstrated promising preclinical activity against MDR and biofilm-associated infections, although robust clinical evidence remains limited. Continued surveillance, antimicrobial stewardship, pharmacokinetic/pharmacodynamic-guided therapy, and well-designed prospective clinical studies are essential to optimize the management of S. maltophilia infections and facilitate the translation of emerging therapeutic strategies into clinical practice.
Marine-derived natural products are promising sources of new anticancer agents, but the bioactivity of Portuguese macroalgae and halophytes remains insufficiently explored. This study aimed to identify extracts with selective antiproliferative activity against breast cancer cells using environmentally sustainable extraction methods. Crude extracts were prepared by ultrasound-assisted extraction with water or 10% dimethyl sulfoxide (DMSO) and screened in human MCF-7 breast adenocarcinoma cells using the sulforhodamine B (SRB) assay after 48 h of exposure. Active extracts were subsequently evaluated in non-tumorigenic MCF-12A breast epithelial cells, and half-maximal effective concentrations (EC50) were determined from concentration–response curves. Of 48 crude extracts screened, 21 inhibited MCF-7 cell proliferation in a concentration-dependent manner and were selected for further evaluation. Three extracts derived from the brown macroalgae Bifurcaria bifurcata II and Ericaria selaginoides—two aqueous and one DMSO-based—showed selective activity against MCF-7 cells without detectable inhibition of MCF-12A cells within the corresponding concentration ranges. These findings identify Portuguese macroalgae as promising sources of selectively active anticancer metabolites and support further phytochemical characterization and mechanistic investigation using sustainable bioprospecting approaches.
Esophageal cancer is biologically heterogeneous, and conventional clinicopathological staging incompletely captures variation in patient outcomes. This study aimed to determine whether histopathology-derived features associated with phosphoinositide metabolism could support exploratory prognostic stratification and identify biologically interpretable epithelial states. Diagnostic whole-slide images and bulk transcriptomic data from The Cancer Genome Atlas Esophageal Carcinoma (TCGA-ESCA) cohort were integrated with single-cell RNA-sequencing data. Tissue-rich image tiles underwent manual quality control, deep features were extracted using an ImageNet-pretrained ResNet-50 model, and phosphoinositide metabolism activity was quantified by single-sample gene set enrichment analysis (ssGSEA). Associated image features were evaluated using machine-learning survival models, followed by clinicopathological adjustment and multimodal molecular characterization. The selected random survival forest plus gradient boosting machine model stratified overall survival in the training and internal validation cohorts, with concordance indices of 0.762 and 0.723, respectively. The standardized pathology-derived risk score remained associated with overall survival after adjustment for age, sex, histological subtype, and pathological stage (hazard ratio, 2.37; 95% confidence interval, 1.60–3.50). However, calibration was imperfect, and performance estimates may be optimistic because the internal validation cohort informed model selection. Most initial pathology–phosphoinositide metabolism associations were attenuated after adjustment for histological subtype. Multimodal analyses prioritized PLEKHA6 as a candidate pathology-associated epithelial-state marker rather than a subtype-independent prognostic biomarker. PLEKHA6-positive epithelial cells exhibited inflammatory and microenvironment-related transcriptional programs, predicted midkine- and macrophage migration inhibitory factor-related communication features, and stronger inferred copy-number variation-like signals. Higher tumor-level PLEKHA6 expression was also associated with distinct inferred immune, metabolic, and predicted drug-response profiles, although malignant-cell identity was not established. Because the analyzed tiles were not derived from pathologist-annotated malignant regions, the image signal represents composite diagnostic-slide tissue context. These findings identify an exploratory pathology-derived prognostic signal and a PLEKHA6-associated epithelial state in esophageal cancer that require independent, subtype-specific, pathological, and experimental validation.
Chronic kidney disease (CKD) is frequently accompanied by left ventricular remodeling, a major contributor to cardiovascular morbidity and mortality, while vitamin D deficiency is highly prevalent in this population. This systematic review and meta-analysis aimed to assess the association between vitamin D status and left ventricular remodeling and to evaluate whether vitamin D supplementation improves remodeling-related cardiac parameters in adults with CKD. Eight databases were searched through December 25, 2024, for observational and interventional studies reporting left ventricular mass index (LVMI), left ventricular ejection fraction (LVEF), or interventricular septal thickness (IVST). Sixteen studies were included: seven observational studies involving 1,487 participants and nine interventional studies involving 581 participants. In observational studies, higher vitamin D levels were associated with lower LVMI (r = −0.41, 95% confidence interval [CI]: −0.64 to −0.11), although heterogeneity was substantial and the certainty of evidence was very low. Vitamin D supplementation did not significantly improve LVMI (standardized mean difference [SMD] = −0.20, 95% CI: −0.62 to 0.22) or LVEF (SMD = 0.57, 95% CI: −0.32 to 1.46). A possible reduction in IVST was observed (SMD = −0.68, 95% CI: −1.32 to −0.05), but this finding was based on limited and inconsistent evidence. Overall, higher vitamin D status was associated with lower LVMI, whereas current interventional evidence does not establish that vitamin D supplementation improves left ventricular remodeling in CKD. Larger, adequately powered, long-term randomized trials are needed to determine whether specific CKD populations may derive cardiovascular structural benefit from vitamin D therapy.
Altered amino acid metabolism may reflect the acute metabolic response to atherosclerotic cardiovascular disease, but its association with acute myocardial infarction (AMI) remains unclear. This study aimed to characterize serum amino acid profiles in patients with AMI and compare non-ST-segment elevation myocardial infarction (NSTEMI) with ST-segment elevation myocardial infarction (STEMI). In this retrospective cross-sectional study, serum samples from age-comparable controls without symptoms of coronary artery disease (CAD) and 60 patients with AMI, including 35 with NSTEMI and 25 with STEMI, were analyzed using nuclear magnetic resonance (NMR) spectroscopy. Metabolic differences were evaluated using principal component analysis (PCA), orthogonal projection to latent structures discriminant analysis (O-PLS-DA), univariate testing, and logistic regression. Thirty-one serum metabolites were identified, and validated O-PLS-DA models differentiated controls, NSTEMI, and STEMI. Compared with controls, patients with AMI had significantly lower serum concentrations of isoleucine, valine, alanine, leucine, glutamine, tyrosine, histidine, and phenylalanine. All these amino acids except glutamine were also significantly lower in STEMI than in NSTEMI. Reduced isoleucine, valine, alanine, leucine, tyrosine, histidine, and phenylalanine were consistently associated with both AMI and STEMI, whereas reduced glutamine was associated with AMI only. These findings demonstrate distinct serum amino acid alterations across AMI phenotypes and support their further investigation as exploratory indicators of acute metabolic shifts. Prospective longitudinal studies are required to determine their temporal, prognostic, and mechanistic relevance.
Anterior cruciate ligament (ACL) reconstruction is commonly recommended for highly active patients, but interest in biologically augmented nonoperative treatment is increasing. This study evaluated clinical and imaging outcomes and explored factors associated with success after platelet-rich plasma (PRP)-assisted nonoperative treatment of acute ACL injuries. This retrospective case series included 40 highly active patients with magnetic resonance imaging (MRI)-confirmed ACL injuries treated within 6 weeks of injury between 2019 and 2023. PRP injections were combined with bracing and staged rehabilitation, and patients were followed for at least 18 months. Treatment success required restored ACL continuity on MRI, a pivot-shift grade ≤1, return to the preinjury sports level, and no major adverse events or subsequent surgery. ACL continuity was restored in 38 patients (95.0%), and 33 (82.5%) met all success criteria. In exploratory analyses, success was more frequent in patients aged ≥30 years than in younger patients (95.2% vs 68.4%, p = 0.027), in partial than complete tears (100.0% vs 66.7%, p = 0.019), and in femoral-side than mid-substance tears (92.6% vs 61.5%, p = 0.029). All seven treatment failures involved complete tears, and five involved mid-substance tears. However, none of these factors was independently associated with treatment failure in exploratory Firth penalized logistic regression. PRP-assisted nonoperative management yielded favorable clinical and MRI outcomes in this selected cohort. These findings are hypothesis-generating, and the independent contribution of PRP cannot be determined because of the retrospective uncontrolled design and limited number of treatment failures.
