Biomedicines doi: 10.3390/biomedicines14040901

Authors: Mai S. Sater Hayder A. Giha

Background/Objectives: Chronic metabolic acidosis (CMA) is a mild, persistent acid–base imbalance characterized by low serum bicarbonate and urinary pH and is common in chronic illness, aging, and metabolic disorders such as type 2 diabetes (T2D). This review highlights the critical, yet often overlooked, role of CMA in T2D (CMAD) and its contribution to disease pathophysiology. Methods: We conducted a comprehensive review of the systemic impacts of CMA, from molecular mechanisms to organ-specific dysfunction. The analysis covers physiological pH dynamics in intracellular (IC) and extracellular (EC) fluids and explores their effects on cellular processes, including the cell cycle and apoptosis. Results: At the molecular level, acidosis significantly alters enzyme kinetics, macromolecule metabolism, and ion conductance. Cell-level analysis shows that pH shifts impact proliferation and programmed cell death. Systemically, the manifestations of CMA align closely with T2D features in vital organs, including the pancreas, liver, skeletal muscle, adipose tissue, and the renal, nervous, and immune systems. Our findings indicate that the pathophysiological landscape of T2D largely mirrors the biological effects of chronic acidosis. Conclusions: The alignment between the effects of CMA and the clinical features of T2D suggests that T2D pathophysiology is worth revisiting through the lens of CMAD. This perspective is further supported by therapeutic interventions showing preliminary efficacy signals in limited studies of acid-neutralization in managing T2D symptoms and progression.

Biomedicines doi: 10.3390/biomedicines14040900

Authors: Osama Al-Bataineh Rula Abdallat Ausilah Alfraihat

Background: Noninvasive transdermal insulin delivery using ultrasound technology has gained attention for improving the glycemic control of insulin-dependent patients. Methods: Indirect functional comparison and evaluation of insulin dosage, between noninvasive ultrasound-mediated transdermal delivery and needle injection methods, was achieved utilizing in vivo blood glucose measurements of temporary hyperglycemic rabbits. Nine rabbits were divided into three groups: (i) untreated control, (ii) subcutaneous injection and (iii) ultrasound-mediated transdermal delivery. Animals were anesthetized using a combination of ketamine hydrochloride and sodium xylazine to produce temporary hyperglycemic rabbits during the experiments. The rabbits in the control group did not receive insulin, while the animals in the ultrasound group received insulin transdermally for 10 min utilizing a customized single-element piston-shaped ultrasound transducer operated by multi-frequency electrical signals from 100 to 200 kHz. Rabbits in the direct subcutaneous injection group were anesthetized and injected with 0.25 units/kg of insulin. Results: With an initial blood glucose baseline level of 228.7 ± 13.1 (mg/dL) for all rabbits, the in vivo results of control group showed an increase above the baseline by 129.7 ± 27.3 (mg/dL) at the end of the in vivo experimental period (80 min). However, the ultrasound-mediated delivery and subcutaneous injection groups showed noticeable statistically significant percentage reductions in blood glucose levels by 43.9 ± 5.4 and 42.7 ± 6.6, respectively, compared to the control group by the end of the in vivo experiments. Conclusions: In vivo glucose response results confirmed that piston-shaped ultrasound transducers achieved indirectly similar insulin dosage delivery by ultrasound energy for tested animals with no statistically significant differences once compared to the results of the subcutaneous needle injection group.

Biomedicines doi: 10.3390/biomedicines14040899

Authors: Morgan Szalay Ian Seguel Strange Kyle Banwell Sophia Campana Adina Dass Sereen Hej-Ali Halima Mohamed Sahar Khan

Clonal hematopoiesis of indeterminate potential (CHIP) is the clonal expansion of somatically mutated hematopoietic stem cells (HSCs) in the bone marrow. CHIP mutations are relatively common in multiple myeloma (MM) and have been identified as potential biomarkers for poorer survival outcomes. MM is a hematological malignancy that, despite treatment advances, remains aggressive and incurable for many patients. The potential impact of CHIP mutations on the outcomes of MM treatments has been the topic of several recent studies, yet both the magnitude and the modality by which CHIP exerts its negative effects on treatment and disease progression remain to be fully elucidated. Evidence suggests that CHIP mutations may contribute to inferior survival and treatment tolerances, as well as contribute to greater treatment toxicity and related frailty. In this review, we synthesize and discuss the available literature to provide an updated understanding of the complex role that CHIP plays in altering the MM microenvironment, and the resulting impact on standard MM treatments, autologous stem cell transplant (ASCT) and B-cell maturation antigen (BCMA)-targeted therapy/CAR-T, and the important role of immunomodulatory drug (IMiD) maintenance therapy in clinical outcomes.

Biomedicines doi: 10.3390/biomedicines14040898

Authors: Karim Gasmi Moez Krichen Afrah Alanazi Sahar Almenwer Sarah Almaghrabi Samia Yahyaoui

Context: In numerous low- and middle-income nations, malaria remains a significant issue due to the challenges associated with diagnosing it through thin blood smears. The appearance of images can vary significantly depending on the microscope type, magnification, lighting conditions, slide preparation methods, and staining techniques. Due to the delicate morphology of parasites, false negatives might adversely affect patient care. Objective: To achieve optimal outcomes from validation, it is essential to construct a robust and easily replicable process. This pipeline should integrate the optimal elements of classical machine learning and end-to-end deep learning, enhance reliability by pairwise ensembling, and select ensemble weights in a logical, data-driven manner. Method: To achieve our objective, we propose two tracks. The initial track encompasses real-time augmentation, convolution-based feature extraction, and the training of calibrated classical classifiers. The second module focuses on training many convolutional networks from inception to completion. Subsequently, we construct paired ensembles and employ a hybrid methodology to select convex weights for combining the findings. This method initially evaluates a set of candidate weights and then refines them to maximise validation accuracy. Results: The precision of the two-track architecture consistently improves, transitioning from conventional baselines to end-to-end models. Optimal and consistent enhancements are achieved through weighted ensembling. Utilising optimised fusion reduces the incidence of false negatives for subtle parasites and false positives caused by staining artefacts. This yields an accuracy of 96.35% on the reserved data and reduced variance across folds. Conclusions: The integration of augmentation, multiple modelling tracks, and optimal pairwise ensembling yields the highest accuracy in categorising malaria smears. It facilitates further enhancements by incorporating supplementary models, multi-class extensions, and operating-point calibration.

Biomedicines doi: 10.3390/biomedicines14040897

Authors: Angela Gyamfi William A. Cisneros Priyadharshini Manikandan Christopher A. Subi-Kasozi Theodore R. Cummins James A. Marrs

Background/Objectives: Despite the decades-old ban on lead in fuel, plumbing, consumer goods, industrial processes, and various materials, it remains a public health threat due to its persistent nature. Zebrafish (Danio rerio) are highly effective for modeling several disorders, including those affecting neurological and behavioral functions, and are well-suited for assessing the impact of environmental toxins like lead. This study aimed to investigate the neurodevelopmental effects of embryonic lead exposure using the zebrafish model system. Methods: Embryos were exposed to lead acetate (PbAc) at concentrations ranging from 0.3 to 0.7 µg/mL using an exposure window of 6 to 48 h post-fertilization (hpf). Results: PbAc exposure produced sublethal teratogenic effects in a subset of larvae across concentrations, including tail and spinal deformities, craniofacial abnormalities, and uninflated swim bladder observed at 7 dpf. At 3 days post-fertilization (dpf), spontaneous circle swimming behavior suspected to be seizure-like was observed in the lead-exposed larvae and was more pronounced under light conditions in a dose-dependent manner. Electrophysiological recordings confirmed that larvae exhibiting circle swimming behavior had heightened neural activity, indicating a potential seizure-like phenotype driven by lead exposure. Conclusions: Our findings suggest that embryonic lead exposure leads to morphological defects and seizure susceptibility, demonstrating lead’s neurotoxic potential during early development. Seizure-like behaviors occurred in a non-linear concentration-dependent manner with a photosensitive component, and elevated baseline neural excitability was confirmed by local field potential (LFP) recordings.

Biomedicines doi: 10.3390/biomedicines14040896

Authors: Milan Potić Ivan Ignjatović Dragoslav Bašić Ljubomir Dinić Bojan Vučković Nebojša Jovanović Slavica Stojnev

Background/Objectives: Hernia and pelvic organ prolapse often involve defects necessitating reinforcement with synthetic materials. Polypropylene meshes of varying weights are commonly used in abdominal wall repair; however, their impact on tissue response, potentially linked to adverse events during wound healing, remains incompletely understood. This study aimed to evaluate and compare the tissue response to lightweight polypropylene (LWPP) and heavyweight polypropylene (HWPP) grafts used for abdominal wall defect closure in Wistar rats. Methods: Abdominal wall defects were surgically created in male Wistar rats and repaired using either LWPP (43 g/m2) or HWPP (76 g/m2) mesh. Three weeks post-implantation, tissue response and wound closure were evaluated across different phases by assessing expression of CD68, CD56, GATA-3, CD138, vimentin, α-smooth muscle actin, and collagen deposition. Results: HWPP promoted a more pronounced foreign body response, as evidenced by increased CD68, CD56, and CD138 expression, while LWPP improved tissue compliance, enhanced fibroblast migration, and increased vimentin-associated cellular activity. Conclusions: Three weeks post-implantation, HWPP was found to induce a stronger inflammatory response, whereas LWPP implantation was associated with increased vimentin expression, indicating differences in the balance between inflammation and extracellular matrix remodelling during early wound healing.

Biomedicines doi: 10.3390/biomedicines14040895

Authors: Li Wei Chunchen Gao Hongyan Qin

Metabolic dysfunction-associated (non-alcoholic) steatotic liver disease (MASLD) is a chronic inflammatory liver disorder characterized by excessive hepatic lipid accumulation. Its progressive subtype, metabolic dysfunction-associated (non-alcoholic) steatohepatitis (MASH), is featured by enhanced inflammation and liver injury. Some MASH cases are accompanied by hepatic fibrosis, which may progress to cirrhosis and hepatocellular carcinoma (HCC). MASLD is also associated with comorbidities such as cardiovascular disease and chronic kidney disease. To date, only Resmetirom has been approved by the FDA for MASH treatment, highlighting the urgency of investigating MASH pathogenesis and developing effective therapeutic agents. Establishment of experimental animal models which can mimic the clinical symptom of MASLD are fundamental to explore therapeutic targets and advance clinical drugs development. Therefore, this review focus on the pathological features of MASLD/MASH and comprehensively summarizes the current MASH-related mouse models, which can be useful for researchers to select appropriate models in order to explore the underlying mechanisms and dig novel targets for MASH treatment.

Biomedicines doi: 10.3390/biomedicines14040894

Authors: Botond-István Kiss Daniela Tatiana Sala Renáta Moriczi Szabolcs-Attila Gábor Árpád Török Gabriel-Mircea Muresan Tivadar Bara Márton István Dénes Szilárd-Leó Kiss Szilárd-Leó Kiss Orsolya Kiss-Toth Radu-Mircea Neagoe

Background: Major morbidity and mortality remain important concerns after colorectal cancer (CRC) surgery. Cancer-related sarcopenia and heightened systemic inflammation may increase the risk of early postoperative major complications. Methods: In this retrospective single-center study, we analyzed 190 patients undergoing major CRC surgery. Skeletal muscle gauge (SMG) and the pan-immune-inflammation value (PIV) were assessed as preoperative risk markers, and 30-day major complications were evaluated. Results: Low SMG was strongly associated with major complications (OR 6.50, 95% CI 3.24–13.05; p < 0.001), and high PIV was also associated with increased risk (OR 3.51, 95% CI 1.77–6.99; p < 0.001). In multivariable analysis adjusting for age, surgical urgency, and procedure type, low SMG and emergency surgery remained independent predictors of 30-day major complications. The highest-risk phenotype (high PIV/low SMG; n = 23) had a major complication rate of 78.3% (18/23) (p < 0.001). A clinical model including age, urgency, and procedure type yielded an AUC of 0.739 (95% CI 0.661–0.816). Adding low SMG improved discrimination (AUC 0.784, 95% CI 0.711–0.857), with only a small additional increase after adding high PIV (AUC 0.791, 95% CI 0.717–0.864). Conclusions: Preoperative low SMG was independently associated with 30-day major complications after CRC surgery, while PIV provided complementary risk-stratification value. The combined high-PIV/low-SMG phenotype identified patients with particularly high postoperative risk.

Biomedicines doi: 10.3390/biomedicines14040893

Authors: Guillermo Valencia Zaida Morante Yomali Ferreyra Rosario Jacome Patricia Rioja Alexandra Saavedra Silvia Neciosup Tatiana Vidaurre Henry L. Gómez

Background: Hormone receptor-positive/HER2-negative (HR+/HER2−) early breast cancer (EBC) presents a persistent risk of relapse, even beyond 5 years, driving the need for adjuvant intensification strategies. This review analyzes the clinical evidence for CDK4/6 inhibitors (CDK4/6i) in the adjuvant setting. This evidence is then integrated with molecular findings to support the concept of the “carry-over” effect, which is understood as a lasting benefit that persists after the end of active treatment, reflected by a sustained separation of invasive disease-free survival (iDFS) curves during follow-up. Relevant Sections: The main adjuvant trials in EBC are reviewed, with consideration of the “carry-over” effect. Emerging biomarkers and the impact of financial toxicity are also described. Results: PALLAS did not demonstrate a clear on-treatment or post-treatment benefit, whereas PENELOPE-B suggested, at most, a transient early advantage that was not maintained with longer follow-up; therefore, neither trial provides convincing evidence of a durable “carry-over” effect. In contrast, monarchE (abemaciclib) and NATALEE (ribociclib) showed significant improvements in iDFS and, in the case of abemaciclib, a signal of benefit in overall survival, supporting the existence of a clinically relevant post-treatment effect. Conclusions: From a biological perspective, the review proposes that the “carry-over” effect should not be considered a uniform class effect, but rather the result of a sequence of events modulated by pharmacological selectivity (CDK4 vs. CDK6 and additional targets), the induction of cellular senescence, and immunomodulatory effects that could favor the control of micrometastases. In addition, elements that influence interpretation and the need to optimize adherence and toxicity management to “materialize” the benefit in a potentially curable context are discussed.

Biomedicines doi: 10.3390/biomedicines14040892

Authors: Xiangsen Zou Peng Song Shicong Song Guowei Zhang Wang Xiao Tingkang Yang Lin Zhou Yixiong Lin

Background: Pancreatic adenocarcinoma (PAAD) is a highly malignant cancer posing severe clinical challenges. Although the dual role of pyroptosis in tumor progression is increasingly recognized, the prognostic value of its molecular heterogeneity in PAAD remains underexplored. Methods: We integrated multi-omics data and applied interpretable machine learning to construct a predictive framework centered on pyroptosis heterogeneity. Using non-negative matrix factorization (NMF) on pyroptosis-related genes (PRGs), patients were classified into distinct molecular subtypes. Evaluating 117 machine learning combinations, we employed random survival forest (RSF) to build the final model, followed by comprehensive internal and external validation. SHapley Additive exPlanations (SHAP) analysis provided global and local interpretability. Clinical potential was assessed via nomogram, drug sensitivity prediction, single-cell analysis, and immunohistochemical validation. Results: We identified two biologically distinct pyroptosis subtypes and developed a ten-gene pyroptosis subtype-associated gene signature (PSAGS). PSAGS demonstrated robust performance across training, test, and multiple external validation cohorts, outperforming most published models. Multivariate analysis confirmed its independent prognostic value, and a PSAGS-based nomogram exhibited clinical utility. PSAGS-stratified subgroups showed differential responses to immunotherapy, chemotherapy, and targeted agents. Single-cell analysis revealed cell type-specific links between PSAGS scores and pyroptosis activity, indicating that high-PSAGS malignant cells foster an immunosuppressive microenvironment through extracellular matrix (ECM)-mediated signaling. Protein-level validation confirmed upregulation of signature genes in PAAD tissues. Conclusions: This work presents a biologically reliable prognostic model for personalized PAAD management and elucidates how pyroptosis heterogeneity drives tumor progression through cellular interactions.

Biomedicines doi: 10.3390/biomedicines14040891

Authors: Pier-Valerio Mari Alberto Ricci Angelo Coppola Davide Onofrio Fontana David Selvaggio Lorenzo Carriera Simone Ielo Matteo Siciliano Loreta Di Michele Veronica Ojetti

Background: Airway mucus plugging is a key but long-overlooked mechanism of persistent airflow obstruction in both asthma and chronic obstructive pulmonary disease (COPD). Type 2 (T2) cytokines, particularly interleukin (IL)-4 and IL-13, drive goblet cell metaplasia, MUC5AC overexpression, and impaired mucociliary clearance, while eosinophil-derived products increase mucus viscosity and promote plug persistence. Methods: A comprehensive narrative review was conducted by searching PubMed and ClinicalTrials.gov databases from inception to February 2026. Search terms included “mucus plugs,” “mucus plugging,” “biologics,” “dupilumab,” “tezepelumab,” “mepolizumab,” “benralizumab,” “IL-4,” “IL-13,” “MUC5AC,” “quantitative CT,” “functional respiratory imaging,” “asthma,” and “COPD.” Studies were included if they reported original data or systematic evidence on mucus plug quantification, biologic-mediated changes in mucus plug scores, or imaging modalities for mucus assessment in asthma or COPD. Editorials, case reports with fewer than three patients, and studies not available in English were excluded. Two authors (P.-V.M. and A.C.) independently screened titles and abstracts; discrepancies were resolved by consensus. Randomized controlled trials, observational studies, and preclinical studies evaluating mucus plug outcomes and T2-targeted therapies were included. Reference lists of retrieved articles were hand-searched for additional relevant publications. Results: A recent systematic review identified multiple randomized controlled trials and observational studies that showed CT-assessed mucus plug scores go down with biologic therapies targeting the T2 pathway in asthma. Observational data extend this evidence to anti-IL-5/IL-5Rα agents. The VESTIGE trial provided the first functional respiratory imaging evidence of mucus plug resolution with dupilumab. In COPD, the BOREAS/NOTUS and MATINEE trials established the efficacy of dupilumab and mepolizumab in eosinophilic phenotypes; however, differences in inclusion criteria—particularly regarding FeNO thresholds and prior exacerbation burden—may explain divergent effects on lung function endpoints. Mucus plug outcomes have not been evaluated in COPD biologic trials. Quantitative imaging modalities, including HRCT mucus plug scoring, functional respiratory imaging, and hyperpolarized gas MRI, now enable objective assessment of mucus burden. Conclusions: Mucus plugging meets the definition of a treatable trait: it can be measured with CT scoring, it matters clinically, and it responds to T2 cytokine blockade. Adding mucus plug assessment to routine clinical evaluation, together with mucolytic strategies where needed, could move treatment decisions from empirical to biology-based across the asthma–COPD spectrum. Further studies are needed to confirm that mucus plug scoring works as a biomarker of treatment response in COPD and to test whether combining biologics with mucolytics improves outcomes.

Biomedicines doi: 10.3390/biomedicines14040890

Authors: Anna Sinitskaya Maria Khutornaya Alyona Poddubnyak Maxim Asanov Alexander Kostyunin Alexey Tupikin Marsel Kabilov Maxim Sinitsky

Background: Infective endocarditis (IE) affects both native and prosthetic heart valves, the endocardial surface, as well as cardiac implantable electronic devices. Identifying specific IE biomarkers for its early risk stratification remains crucial, particularly in cases with blood culture-negative endocarditis. Methods: Eleven native heart valves obtained from IE and calcific aortic valve disease (CAVD) patients were analyzed. Immunohistochemical analysis of a pan-leukocyte marker (CD45), macrophage marker (CD68), T-lymphocyte marker (CD3), B-lymphocyte marker (CD19), neutrophil myeloperoxidase (MPO), and marker of vascular endothelial cells (CD31) was performed. Differentially expressed genes (DEGs) were identified by whole-transcriptome sequencing; proteomic profiling was performed by dot-blotting. Results: The immunophenotyping demonstrates the infiltration of macrophages and neutrophils, as well as occasional T-lymphocytes in the IE-affected aortic valves, and the CAVD-affected heart valves were characterized by the absence of neutrophils. For the whole-transcriptome sequencing, 157 DEGs were identified: 124 DEGs were upregulated, and 33 genes were downregulated in the IE-affected heart valves compared to the CAVD-affected ones. According to the dot-blotting, 35 cytokines were identified in the studied heart valves, but only 21 molecules were expressed in both IE and CAVD-affected heart valves. Analysis of proteases and their inhibitors allowed the identification of 13 protease molecules and 18 enzyme inhibitor molecules in all examined heart valves. Conclusions: The results of the present study can help to improve our understanding of the IE pathogenesis. In addition, we identified the candidate cellular and molecular-genetic features of IE-affected native heart valves.

Biomedicines doi: 10.3390/biomedicines14040889

Authors: Neal G. Simon Michael J. Brownstein Karen E. Anderson Shi-fang Lu Hilda T. Maibach

Background: Neurodegenerative diseases are a major health problem, and the neuropsychiatric symptoms seen in these diseases adversely impact the lives of patients, families, and caregivers. Inappropriate aggressive behavior is a highly disruptive symptom and a leading cause of institutionalization. There are no approved drugs specifically for the treatment of problematic aggression, and the off-label use of antipsychotics has limited benefit with significant side effects and safety risks. This review discusses dysregulated arginine vasopressin (AVP) signaling in fear–threat circuitry as a key driver of inappropriate aggression. Because the AVP 1a receptor (V1aR) is the dominant subtype in the CNS, the selective antagonism of this receptor represents a well-rationalized target for the treatment of aggression across neurodegenerative, psychiatric, and neurodevelopmental disorders. Objectives: Our goal was to summarize the basis for using V1aR antagonists as a treatment for irritability and aggressive behavior. We describe its discovery, biosynthesis, receptor pharmacology, and CNS distribution, emphasizing V1aR localization in central fear–threat circuits. Translational evidence from animal studies, pharmacological neuroimaging, and lesion network mapping is presented. These data support the suggestion that heightened vasopressinergic tone biases socioemotional information processing toward negative valence, increasing threat sensitivity and the likelihood of inappropriate aggressive responses. Emerging clinical data support this framework. Highly selective, CNS-penetrant V1aR antagonists reduced aggressive behavior and had an excellent safety profile in phase 2 studies in Huntington’s disease and intermittent explosive disorder, with efficacy signals across caregiver-reported, clinician-rated, and incident-based measures. Furthermore, pharmacological neuroimaging showed that V1aR antagonism normalizes AVP-induced alterations in activity within fear–threat circuitry. Conclusions and Future Directions: Preclinical, translational, and clinical findings to date support V1aR antagonism as a promising strategy for treating pathological aggression across disorders. Additional experimental medicine studies and clinical trials are needed to conclusively establish efficacy in various disease populations, and we note the need for improved trial designs and analytical methods as part of the development process.

Biomedicines doi: 10.3390/biomedicines14040888

Authors: Joana Caetano Pedro de la Villa Polo José Delgado Alves Luis Monteiro Rodrigues

Background: Reactive hyperemia (RH) is used to assess microcirculatory function in vivo and has traditionally been interpreted as a local, ischemia-driven vasodilatory response following arterial occlusion. However, perfusion changes consistently observed in contralateral, non-challenged limbs question the exclusively local nature of RH. Objective: This study aimed to characterize reactive hyperemic responses elicited by subsystolic cuff pressures, below arterial occlusion pressure (AOP), and to investigate their effects on glabrous and non-glabrous skin microcirculation and on global hemodynamics. Methods: Seven healthy women underwent a standardized protocol consisting of baseline stabilization, a 2 min subsystolic cuff inflation (70–80% of resting AOP) in one arm, and a recovery period. Microvascular perfusion was simultaneously assessed in both hands using laser Doppler flowmetry (LDF) on glabrous skin and polarized light spectroscopy (PSp) on non-glabrous dorsal skin. Hemodynamic indicators were continuously monitored using CNAP (Continuous Non-invasive Arterial Pressure) technology. Ipsilateral and contralateral responses were compared across experimental phases. Results: Subsystolic cuff inflation induced significant perfusion changes not only in the challenged limb but also in the contralateral limb, despite the absence of a complete arterial occlusion. Conclusions: These findings confirm the adaptive nature of RH emphasizing the major role for the sympathetic nervous system in glabrous skin. In glabrous (palmar) skin, a similar perfusion profile is shown in both hands but significant differences could only be found in the ipsilateral hand. In contrast, non-glabrous (dorsal) skin demonstrated region-specific increases in perfusion, again evident in the ipsilateral hand, suggesting venous stasis. No changes in global hemodynamic variables were observed throughout the protocol. Further studies in larger, more diverse populations are needed to confirm these observations and refine the mechanistic understanding of reactive hyperemia.

Biomedicines doi: 10.3390/biomedicines14040887

Authors: Wen Yang Qi He Zhen Sun Xiaochang Zhang Qingyu Li Changdong Zhou Yuke Cui Zhenqiao Wei Jingqi Shi Chenhui Wang Yuanyuan Jiao Liang Guo Yaling Xing Shengqi Wang

Background/Objectives: Influenza A virus (IAV) infection triggers robust inflammation and acute lung injury. Andrographolide, a primary active compound from Andrographis paniculata, can mitigate IAV-induced inflammation; however, its precise mechanisms remain poorly elucidated. This study aimed to define its host-directed protective effects and molecular mechanisms. Methods: We used a lethal IAV (H1N1, PR8) model in BALB/c mice and infected A549 cells. Survival, lung pathology, cytokines, and viral titers were measured. Lung RNA sequencing identified dysregulated signaling pathways. PI3K/AKT and pyroptosis pro-teins were analyzed by Western blot. The PI3K/AKT axis was functionally validated with the AKT inhibitor in vivo and AKT1 siRNA in vitro. Results: Andrographolide improved survival, attenuated body weight loss, and reduced lung pathology and inflammatory cytokine levels in IAV-infected mice, without exhibiting direct antiviral activity. Consistent with the in vivo findings, andrographolide enhanced cell viability and suppressed cytokine secretion in infected cells. RNA sequencing revealed marked upregulation of the PI3K/AKT signaling pathway in the lungs of treated mice, as confirmed by increased PI3K and AKT phosphorylation. Furthermore, andrographolide downregulated the expression of key pyroptosis-executing proteins, including cleaved caspase-3 and the gasdermin E (GSDME) N-terminal fragment. These protective effects were substantially abrogated by an AKT inhibitor and AKT1 siRNA. Conclusions: These findings reveal a novel host-directed mechanism by which andrographolide alleviates IAV-induced immunopathology by activating the PI3K/AKT pathway, thereby suppressing caspase-3/GSDME-dependent pyroptosis. Thus, this axis represents a promising target for controlling excessive inflammation in severe influenza.

Biomedicines doi: 10.3390/biomedicines14040886

Authors: Ahmed Mansour Al Rajeh

The lung–kidney axis forms an important physiologically integrated system which controls multiple essential functions of the body. An important observation of this interaction is tissue oxygenation and erythropoiesis, a vital process that involves erythropoietin (EPO) release by the kidney to bring red cell production into the bone, while pulmonary gas exchange ensures adequate oxygen delivery to the cells. Subsequently, the lung–kidney activation of the renin angiotensin system (RAS) influences vascular tone, blood pressure, and tissue perfusion, influencing the delivery of oxygen and the body’s requirement for erythropoietin. Additionally, beyond oxygen sensing, studies have evidenced the role of hypoxia-inducible factors (HIFs), inflammatory mediators, endothelial signaling pathways and iron availability. These modulate erythropoietin production, which enhances the process of erythropoiesis and arterial oxygen balance. Localized variations in renal oxygen levels together with hemodynamic control mechanisms enable the body to produce erythropoietin independently from systemic hypoxia conditions. This concept emerged to include the renal oxygen extraction fraction (OFE) and intrarenal microvascular shunting with perfusion oxygen coupling in governing EPO production. The present review refines the traditional knowledge to further expand our understanding of the lung–kidney axis regulating the process of erythropoiesis and arterial oxygen content. The integrative framework demonstrates that pulmonary arterial oxygenation and renal oxygen sensing together with bone hematopoietic responses operate as a unified system which maintains both oxygen equilibrium and hematopoietic balance throughout the body.

Biomedicines doi: 10.3390/biomedicines14040885

Authors: Sheng-Huei Wang Kuang-Yao Yang Chau-Chyun Sheu Biing-Ru Wu Ming-Cheng Chan Jia-Yih Feng Chia-Min Chen Yi-Cheng Shen Wei-Hsuan Huang Chung-Kan Peng Shih-Ming Huang

Background: End-stage renal disease (ESRD) is an immunocompromised state that confers a high risk of infection. We aimed to integrate bioinformatics analyses with a clinical cohort to explore the association between ESRD and sepsis. Methods: We retrieved transcriptomic data from the Gene Expression Omnibus and used computational tools, including Gene Set Enrichment Analysis, the eXtreme Gradient Boosting algorithm, and Mendelian randomization, to characterize gene expression changes, biological pathways, and genetic features in ESRD and sepsis. A multicenter retrospective cohort of patients with sepsis due to carbapenem-resistant Acinetobacter baumannii (CRAB) pneumonia in intensive care units (ICUs) was used to compare clinical presentation and outcomes between patients with and without ESRD. Results: Differential gene expression analysis showed widespread transcriptomic dysregulation in ESRD, and functional enrichment analysis revealed perturbations in immune signaling and vesicular transport pathways. Both the innate and adaptive immune systems appeared compromised, with marked depletion of lymphoid lineages in ESRD. An XGBoost machine-learning model derived from immune cell enrichment scores demonstrated a similar immune microenvironment in ESRD and sepsis. Mendelian randomization analysis supported an association between genetic variants predisposing to ESRD and an increased risk of sepsis, using genome-wide association study datasets. In the clinical cohort, patients with ESRD had significantly higher Sequential Organ Failure Assessment (SOFA) scores and in-hospital mortality than patients with normal renal function. Conclusions: ESRD shares similar immune microenvironmental features and genetic signatures with sepsis. These shared characteristics may contribute to the greater sepsis severity and poorer outcomes observed in patients with ESRD.

Biomedicines doi: 10.3390/biomedicines14040884

Authors: Aurora Ferro Andrea Segreti Nardi Tetaj Martina Ciancio Simone Pasquale Crispino Riccardo Cricco Chiara Fossati Fabio Pigozzi Francesco Grigioni

In heart failure (HF), elevated blood lactate levels, particularly during exercise or in advanced disease stages, reflect impaired tissue perfusion and altered metabolic regulation. Beyond its traditional role as a marker of anaerobic metabolism, lactate has emerged as a dynamic indicator of metabolic reserve and ventilatory control. This narrative review summarizes current evidence on lactate dynamics at rest and during exercise, highlighting their pathophysiological and clinical relevance. In HF patients, exercise-induced lactate accumulation occurs earlier and at lower workloads, reflecting impaired oxidative capacity and reduced peripheral oxygen utilization. This phenomenon is closely associated with ventilatory inefficiency, as demonstrated by the relationship between lactate levels and the VE/VCO2 slope during cardiopulmonary exercise testing (CPET). Emerging data suggest that lactate is not only a marker of disease severity but also a potential mediator of chemoreflex activation and abnormal ventilatory responses. Furthermore, both pharmacologic and non-pharmacologic interventions may influence lactate production and utilization, supporting its role as a potential tool for therapeutic monitoring. Overall, the integration of lactate assessment, particularly during exercise, into clinical evaluation may provide additional insight into disease mechanisms, improve risk stratification, and contribute to personalized therapeutic optimization in patients with HF.

Biomedicines doi: 10.3390/biomedicines14040883

Authors: Isabella Luisa Walther Karim Mostafa Agreen Horr Sandra Freitag-Wolf Hatim Seoudy Oliver J. Müller Sarah Krutmann Olav Jansen Patrick Langguth

Purpose: Thymic involution, a hallmark of immune ageing, is associated with chronic low-grade inflammation (“inflammaging”) and has been implicated in age-associated inflammatory diseases, including atherosclerosis. This study aimed to evaluate the association between persistent thymus and coronary artery calcification based on the Agatston score. Materials and Methods: In an exploratory effort, we retrospectively analyzed 206 patients aged 40–64 years who underwent ECG-triggered thoracic CT between 2019 and 2024. Coronary artery calcifications were quantified on virtual non-contrast reconstructions using the Agatston score. Thymic tissue was graded on a five-point scale based on the extent of fatty replacement, with higher grades indicating greater thymic preservation. Results: The cohort included 126 men and 80 women. Complete fatty replacement of the thymus (Grade 0) was seen more often in men compared to women (51/126 vs. 18/80; p = 0.011). Linear regression analysis revealed a significant inverse association between thymus grade and coronary Agatston score (Beta (B) = −28.8 (95% CI −45.3 to −12.3); p = 0.001). After adjusting for age and sex, higher thymic grades remained significantly associated with lower coronary Agatston scores (B = −22.2 (95% CI: −41.7 to −2.6); p = 0.03). Further analysis with adjustments for cardiovascular risk factors was not performed. Conclusions: Residual thymic tissue was significantly inversely associated with coronary artery calcification, and this association persisted after adjustment for age and sex. These findings support the hypothetical concept that morphologically detectable thymic remnants may reflect interindividual differences in immune ageing and inflammaging that are associated with age-related inflammatory disease phenotypes. The results of this hypothesis-generating study give incentive to further investigate the nature and strength of these associations in prospective studies.

Biomedicines doi: 10.3390/biomedicines14040882

Authors: Shang-Wun Jhang Liang-Fang Lin Gizem Naz Canko Bill Cheng

Background/Objectives: Macrophage phenotype and function are highly sensitive to environmental cues; however, most in vitro studies rely on 2D culture systems that lack physiologically relevant structural context. The spatial dimensionality can influence immune cell signaling, yet the roles of these cells in regulating macrophage behavior remain incompletely understood. This study aimed to investigate how cultural dimensionality affects the phenotype, signaling, and functional activity of monocyte-derived macrophages. Methods: GFP-expressing THP-1 monocytes were differentiated into M0, M1, and M2 macrophages and cultured either on planar substrates or within 3D matrices composed of Matrigel or type I collagen. Macrophage morphology and viability were monitored. Membrane receptor expression and secreted cytokines were examined and quantified. Functional activity was further assessed through coculture experiments with RFP-expressing MDA-MB-231 breast cancer cells. Results: Compared with 2D culture, 3D environments induced distinct morphological and viability changes in macrophages. Collagen matrices supported sustained growth, subtype-specific morphologies, and enhanced functional activity, whereas Matrigel promoted aggregation and reduced viability. Core lineage markers remained stable across conditions, but activation-associated receptors and cytokine profiles were strongly influenced by dimensionality. 3D culture enhanced TNF-α expression and altered serglycin glycosylation patterns. In coculture assays, macrophage effects on tumor cell growth depended on polarization state and were more pronounced in 3D systems. Conclusions: These findings demonstrate that culture dimensionality and ECM composition are key regulators of macrophage phenotype and function. Collagen-based 3D systems better reproduce physiologically relevant macrophage behaviors than conventional 2D platforms, highlighting the value of structurally biomimetic models for immunological studies and therapeutic screening.

Biomedicines doi: 10.3390/biomedicines14040881

Authors: Yaling Zhang Ming Li Yanwen Luo Liming Huang Sipei Chen Guisen Li Yi Li Li Wang

Background: Chronic kidney disease (CKD) associated vascular calcification (VC) is a leading cause of cardiovascular mortality, partially driven by osteogenic transdifferentiation of vascular smooth muscle cells (VSMCs). Chaperone-mediated autophagy (CMA) is a selective lysosomal degradation cellular process. However, the precise role and mechanism of CMA in CKD-associated vascular calcification remain unknown. Methods: We studied calcified arteries from CKD patients and rats fed on a high-phosphate diet using histological and ultrastructural methods. VSMCs' calcification was induced by a calcification medium containing high phosphate and calcium. CMA activity was measured by a KFERQ reporter and lysosomal staining. The expression of LAMP2a and HSP90AA1 was knocked down by siRNA, overexpressed by plasmid, and activated by QX77.1. Bioinformatic analysis, protein interaction studies, immunofluorescence and co-immunoprecipitation were performed to investigate the potential mechanism of CMA in VC. Results: The expression of LAMP2a was increased in human calcified radial artery tissues (n = 3, p < 0.05) and rats' calcified aortic tissues (n = 3, p < 0.01), accompanied by lysosomal abnormalities. The activity of CMA was increased during the osteogenic transdifferentiation of VSMCs, as indicated by increased expression of RUNX2 and reduced expression of SM22α (p < 0.05). LAMP2a knockdown attenuated VSMCs’ calcification (p < 0.05), whereas pharmacological activation of CMA aggravated calcification in VSMCs (p < 0.01). Bioinformatic screening identified HSP90AA1 as a candidate involved in CMA in vascular calcification. Elevated HSP90AA1 expression was observed in human calcified radial artery tissues (n = 3, p < 0.01) and rat calcified aortic tissues (n = 3, p < 0.01), which promoted osteogenic transdifferentiation of VSMCs (p < 0.05). HSP90AA1 interacted with LAMP2a and positively regulated its expression (p < 0.01). Conclusions: These findings support an association between CMA activation and CKD vascular calcification. It suggests that HSP90AA1 facilitates vascular calcification in chronic kidney disease involving chaperone-mediated autophagy.

Biomedicines doi: 10.3390/biomedicines14040880

Authors: Anna Tsiakiri Christos Kokkotis Dimitrios Tsiptsios Leonidas Panos Nikolaos Aggelousis Konstantinos Vadikolias Foteini Christidi

Background/Objectives: Minor neurocognitive disorder (minor NCD) represents a heterogeneous and potentially modifiable stage along the continuum from normal aging to dementia, offering a critical window for targeted health promotion interventions. Early identification of individuals at increased risk of progression is essential for implementing preventive strategies that may delay functional decline. This study developed a transparent machine learning (ML) framework to predict diagnostic change from minor to major NCD at 12 and 24 months using baseline demographic, clinical, and multidomain neuropsychological data. Methods: A retrospective cohort of 162 memory clinic patients was analyzed using a rigorously controlled pipeline incorporating nested stratified cross-validation, SMOTE-based imbalance correction, and sequential forward feature selection. Logistic regression, support vector machines (SVMs), and XGBoost were evaluated, with SHapley Additive exPlanations (SHAPs) applied to ensure interpretability. Results: SVM achieved the most balanced predictive performance at both 12 months (accuracy = 0.90) and 24 months (accuracy = 0.81). Short-term progression was primarily driven by subtle multidomain cognitive inefficiencies, while longer-term risk reflected continued cognitive vulnerability modulated by metabolic factors such as diabetes. Conclusions: These findings highlight the potential of explainable ML as a health promotion tool and suggest that explainable ML can uncover clinically meaningful cognitive risk signatures at the earliest stages of NCD. By identifying modifiable systemic contributors alongside cognitive risk profiles, this framework supports precision-oriented preventive strategies and proactive longitudinal monitoring in minor NCD.

Biomedicines doi: 10.3390/biomedicines14040878

Authors: Yildirimcan Demirtas Husnu Cagri Genc Mustafa Ozkaraca Mahmut Sahin Alper Serhat Kumru Atilla Kurt

Objectives: Acute peritonitis remains a critical condition with high mortality rates, further complicated by the rising antibiotic resistance. This study aimed to investigate the therapeutic potential of Chlorella vulgaris (CHL), both alone and in combination with standard antibiotic therapy (SFT), in a rat model of cecal ligation and puncture (CLP)-induced peritonitis. Methods: Seventy (70) male Wistar albino rats were divided into seven groups (n = 10): Control, Peritonitis Control, Low-dose CHL (CHL I) (150 mg/kg), High-dose CHL (CHL II) (300 mg/kg), the standard first-line therapy group (SFT) (Ceftriaxone + Metronidazole), SFT + CHL I, and SFT + CHL II. Following CLP-induced peritonitis, treatments were administered for 7 days. Peritoneal tissues were evaluated histopathologically and immunohistochemically for TNF-α, IL-1β, IL-10, and iNOS expression. Total Antioxidant Status (TAS) and Total Oxidant Stress (TOS) were measured to assess the oxidative stress. Results: Histopathological analysis showed that CLP-induced severe inflammatory damage was significantly reduced in all treatment groups, with the most prominent recovery observed in the SFT + CHL II group. CHL treatment led to a significant decrease in pro-inflammatory markers (TNF-α, IL-1β, iNOS) compared to the peritonitis control group (PC) (p < 0.05). Furthermore, CHL administration significantly improved the oxidative balance by increasing TAS and reducing TOS levels. Conclusions: Chlorella vulgaris exhibits significant anti-inflammatory, antioxidant, and immunomodulatory properties in experimental peritonitis. When used as an adjunct to standard antibiotic therapy, high-dose CHL provides synergistic effects that contribute to limiting tissue damage and controlling systemic inflammation. These findings suggest that CHL may be a promising supportive agent in the clinical management of acute peritonitis.

Biomedicines doi: 10.3390/biomedicines14040879

Authors: Lamprini Vlachodimitropoulou Marios Lampros George A. Alexiou Anastasia K. Zikou Spyridon Voulgaris Paraskevi V. Voulgari

Traumatic brain injury (TBI) is a major cause of death and disability, mainly in persons under 45 years of age and it remains clinically challenging due to its heterogeneous pathophysiology and unpredictable course. Except from the initial mechanical damage, secondary injury —largely driven by neuroinflammation—plays a critical role in outcome and extent of recovery. Cytokines are central mediators of this immune response and have therefore been extensively studied as potential biomarkers for TBI diagnosis, need of imaging and prognosis. Among pro-inflammatory cytokines, IL-1β is rapidly upregulated after TBI and contributes to blood–brain barrier disruption and secondary damage. Furthermore, experimental studies suggest that IL-1 inhibition could be neuroprotective. IL-6 is up to date the most extensively studied cytokine and shows strong associations with injury severity, neuroimaging abnormalities, mortality and long-term functional outcomes across multiple adult and pediatric studies. Nevertheless, results vary depending on the biological compartment and timing. Anti-inflammatory IL-10 levels correlate with injury severity and has shown promise in distinguishing CT-positive from CT-negative mild TBI patients, potentially reducing unnecessary imaging, though findings are inconsistent. Other cytokines, including IL-17, TNF-α, IL-8, IL-9, and IL-15, have been correlated to post-traumatic neuroinflammation and may have diagnostic or prognostic value. Overall, IL-6 and IL-10 currently appear to be the most promising cytokine as biomarkers, however future research should focus on standardized cytokines assessment methods and possible use of multimarker panels.

Biomedicines doi: 10.3390/biomedicines14040877

Authors: Elliott L. Fite Nikhil Sekar Jenish S. Venancius Mina S. Makary

Hepatocellular carcinoma (HCC) remains a major contributor to global cancer mortality, with many patients presenting beyond the bounds of upfront curative therapy (resection/transplant). Locoregional therapies, particularly transarterial chemoembolization (TACE), transarterial embolization (TAE), and transarterial radioembolization (TARE), therefore play an essential role in bridging and downstaging strategies designed to enable curative intent in otherwise ineligible patients. Bridging therapy aims to maintain transplant eligibility and reduce waitlist dropout, whereas downstaging seeks to reduce tumor burden to meet accepted criteria for resection or transplantation. This review synthesizes current evidence on TACE, TAE, and TARE for bridging to resection and transplantation, as well as for downstaging to surgical eligibility, drawing from systematic reviews and cohort studies in the recent literature. We examine modality-specific outcomes, contextualized by tumor biology, liver function, and treatment selection criteria. Comparative effectiveness and the need for standardized outcome measures will be highlighted, reflecting heterogeneity in study endpoints and patient populations. Finally, future directions in personalized locoregional therapy, integration with systemic therapies, and refined conversion strategies will be discussed, with emphasis on the need for consensus in defining treatment success. By integrating evolving clinical evidence with practical application, this review will help clarify the expanding role of locoregional therapies in enabling curative-intent strategies for HCC.

Biomedicines doi: 10.3390/biomedicines14040876

Authors: Filippo Catalani Arianna Pannunzio Emanuele Valeriani Walter Ageno Pasquale Pignatelli Sandor Györik

Background: Cancer and pulmonary circulation disorders represent increasingly intersecting clinical entities. The prevalence of malignancy in patients with pulmonary hypertension (PH), particularly those with chronic thromboembolic pulmonary hypertension (CTEPH), is higher than in the general population. Moreover, cancer and antineoplastic therapies have been implicated in the development of PH through multiple mechanisms. Methods: We performed a systematic review and meta-analysis of the literature focusing on the prevalence of cancer in patients with PH. Mortality incidence and mortality risk were also evaluated for patients with PH with or without cancer. Specific sub-analyses for patients with CTEPH were also performed. Finally, we evaluated the prevalence of PH and its risk of mortality in patients with cancer. Results: Overall, 12 studies including 4402 patients were selected in the quantitative analysis. All the included studies had an observational design. The prevalence of cancer in patients with any PH group was 13% (95% CI: 11–16%); mortality incidence in patients with any PH group and cancer was 41% (95% CI: 26–58%), compared to 10% (95% CI: 1–48%) in those without cancer. The association was even stronger when considering only patients with CTEPH, with a mortality incidence of 4% (95% CI: 2–9%) in those without cancer compared to 19% (95% CI: 8–37%) in patients with cancer (p for difference: < 0.01). Finally, prevalence of any PH group in patients with cancer was 22% (95% CI: 15–31%). Conclusions: We observed a possible correlation between PH and cancer, with a significant impact on mortality in patients with PH, particularly those with CTEPH. This association suggests the need for a close clinical surveillance for early detection of cancer and PH.

Biomedicines doi: 10.3390/biomedicines14040875

Authors: Rodrigo L. Castillo Emilio Jofré-Saldía Daniela Cáceres-Vergara Georgina M. Renard Esteban G. Figueroa

Population aging has markedly increased the burden of cancer in older adults, in whom frailty, sarcopenia, and reduced physiological reserve limit tolerance to treatment and worsen clinical outcomes. Aging is accompanied by progressive functional decline and by biological processes such as cellular senescence, characterized by irreversible cell cycle arrest, chronic low-grade inflammation, and impaired immune surveillance. The accumulation of senescent cells and the persistence of a senescence-associated secretory phenotype contribute to tissue dysfunction and generate a microenvironment that favors tumor initiation and progression. Physical exercise has been associated with attenuation of inflammation, improvements in metabolic and immune function, and with lower levels of senescence-related biomarkers. Although aerobic exercise has been extensively studied in this setting, resistance training holds relevance for older adults due to its capacity to counteract sarcopenia, preserve muscle strength and power, and sustain functional independence. Structured and periodized approaches to resistance exercise may further enhance these benefits by delivering targeted stimuli aligned with age-related physiological deficits. Block strength training (BST), a periodized model that concentrates training adaptations into sequential phases of maximal strength, power, and muscular endurance, has demonstrated consistent improvements in functional performance and reductions in frailty risk in community-dwelling older adults. BST improves physical function. It may also influence biological processes related to aging and cancer; however, mechanistic evidence specific to BST remains to be established. We hypothesized that the exercise in block as a targeted, a structured and physiologically grounded resistance training intervention highlights the potential of BST to promote functional aging and healthy. In the case of cancer biology, and the environment near to tumour, the relationship between aging mechanisms in older adults and controlled exercise effects are currently in advance, but mechanistic trials are still lacking. Finally, we propose a novel training method, structured and personalized, that could impact different clinical outcomes in older patients with cancer.

Biomedicines doi: 10.3390/biomedicines14040874

Authors: Peixin Wu Yue Yin Jinxia Liu Zhenfei Mo Jiabo Ren Xiuqing Ma Zhixin Liang Miaoyu Wang Chunsun Li Liangan Chen

Background/Objectives: High-altitude pulmonary edema (HAPE) remains a serious condition with limited preventive options. This study evaluated the prophylactic protective effects of nebulized human umbilical cord mesenchymal stem cell-derived extracellular vesicles (hUC-MSC-EVs) in a rat model of hypobaric hypoxia-induced lung injury and explored potential mechanistic clues, with a focus on oxidative stress and TEK/Tie2 signaling. Methods: Rats were exposed to hypobaric hypoxia (47 kPa; 9.7% O2) for 72 h and received prophylactic nebulized hUC-MSC-EVs (300 μg/rat). Lung injury was evaluated by histopathology, wet-to-dry ratio, and bronchoalveolar lavage fluid (BALF) protein concentration. Invasive pulmonary function indices were measured using a forced oscillation system. BALF cytokines (TNF-α, IL-6, and IL-10), reactive oxygen species (ROS), and TEK/Tie2 expression in lung tissue were assessed. In addition, transcriptome sequencing (RNA-seq) was performed to characterize global transcriptional changes. N-acetylcysteine (NAC), a classical antioxidant, was included as an auxiliary mechanistic intervention to assess the association of ROS with TEK/Tie2 changes. Results: Compared with hypoxia controls, prophylactic nebulized hUC-MSC-EVs reduced histopathological injury, pulmonary edema, and barrier leakage, and improved pulmonary function indices. hUC-MSC-EV intervention also attenuated inflammatory responses in BALF, with decreased TNF-α and IL-6 and increased IL-10. Hypobaric hypoxia increased ROS accumulation and decreased TEK/Tie2 expression, whereas nebulized hUC-MSC-EVs reduced ROS and partially preserved TEK/Tie2 expression. NAC pretreatment similarly reduced ROS and was accompanied by Tie2 preservation. Conclusions: Prophylactic nebulized hUC-MSC-EVs mitigated hypobaric hypoxia-induced lung injury, accompanied by reduced oxidative stress, improved vascular barrier integrity, and preservation of TEK/Tie2 expression. These findings support nebulized hUC-MSC-EVs as a potential lung-targeted prophylactic strategy for hypobaric hypoxia-induced lung injury and suggest that ROS imbalance may be associated with Tie2 preservation.

Biomedicines doi: 10.3390/biomedicines14040872

Authors: Dmitrii A. Leonov Irina A. Mednova Alexander A. Chernonosov

Background: Untargeted metabolomics enables comprehensive profiling of biological systems, but accurate metabolite annotation remains a critical bottleneck due to incomplete spectral libraries and structural isomerism. The use of in silico annotation tools can increase the coverage of annotated compounds, but it remains unclear whether these tools, in the absence of reference standards, can reliably annotate real-world experimental LC-HRMS data and whether they are sufficient for this task. Methods: This study assesses the performance and limitations of four widely used in silico structure prediction tools (MassFrontier, MetFrag, MS-FINDER, and SIRIUS/CSI:FingerID) when applied to an experimentally acquired feature set previously used to differentiate patients with depressive disorders from healthy controls. To ensure uniform evaluation across tools under realistic but optimized conditions, the quality of MS/MS data was improved using a parallel reaction monitoring method, allowing acquisition of interpretable fragmentation spectra for 26 of the 28 detected features. Results: For most features, all tools were able to suggest structure candidates. However, none of the tools proved sufficient as a standalone solution for reliable metabolite annotation. Due to their different algorithms, each tool had strengths and weaknesses in fragmentation interpretation, candidate generation, and ranking, resulting in incomplete or inconsistent annotations. While the combined application of all four tools provided a substantial improvement in putative annotation over conventional spectral library matching, the in silico structure prediction tools often prioritized chemically implausible, biologically irrelevant, or artifactual candidates. Consequently, manual expert evaluation was required to assess the chemical plausibility and biological relevance of the proposed structures. This ultimately reduced the number of biologically plausible metabolites putatively associated with disease to ten. Conclusions: Overall, these results demonstrate that existing in silico annotation tools can substantially support the annotation of experimental metabolomics data, but are insufficient on their own. Reliable identification of metabolites in complex biological matrices still depends on high-quality MS/MS data acquisition, the combined use of complementary tools, and mandatory post-annotation expert curation.

Biomedicines doi: 10.3390/biomedicines14040873

Authors: Nathan Engel Daniel Oliveira Craig Sykes Amanda P. Schauer Jasmine L. King Thy Le Soumya Rahima Benhabbour Mackenzie Cottrell

Background/Objectives: Women and girls, particularly in sub-Saharan Africa, face high risks for both HIV and unintended pregnancy. Inconsistent condom use underscores the need for new multipurpose prevention technologies (MPTs) that combine HIV pre-exposure prophylaxis (PrEP) and contraception. Long-acting (LA) injectables are especially promising. To this end, an LA cabotegravir (CAB)/medroxyprogesterone acetate (MPA) in situ-forming implant (ISFI) has been developed. We report pharmacokinetic (PK) modeling to characterize CAB and MPA disposition and absorption to support the development of the MPT ISFI. Methods: Female BALB/c mice received single intravenous (IV) or subcutaneous (SQ) bolus doses of CAB or MPA. Sparse plasma samples were collected (~3 mice/timepoint) for PK analysis by LC-MS/MS. Noncompartmental analysis assessed SQ bioavailability. Macroparameterized compartmental PK models were fit to IV data to derive unit impulse responses (UIRs) for each drug. Results: CAB and MPA exhibited 61% and 42% bioavailability, respectively. CAB IV PK was best described by a two-compartment model with macroconstant parameters: A = 16,621 ng/mL, α = 4.52 h−1, B = 30,206 ng/mL, and β = 0.053 h−1. MPA IV PK was also best described by a two-compartment model, with A = 2506 ng/mL, α = 10.5 h−1, B = 439 ng/mL, and β = 0.65 h−1. These values define the UIR for CAB and MPA. Conclusions: Our IV PK modeling framework fully characterizes CAB/MPA disposition in mouse, enabling downstream deconvolution-based estimation of absorption from controlled-release formulations. This provides a foundation for in vitro–in vivo correlation, facilitating preclinical evaluation of long-acting formulations such as ISFIs.

Biomedicines doi: 10.3390/biomedicines14040871

Authors: Angela Vidal Anaïs Y. Kilian Vithusha Vinayahalingam Branislav Zagrapan Janna Pape Tanya Karrer Michael von Wolff

Background: Chronic endometritis (CE) is a subtle, often asymptomatic endometrial inflammation marked by CD138+ plasma cell infiltration and linked to recurrent implantation failure (RIF), recurrent pregnancy loss (RPL), and unexplained infertility. Emerging evidence implicates endometrial microbiome dysbiosis in CE. Objective: To systematically review and conduct meta-analysis on the association between CE and endometrial microbiome alterations and their reproductive implications. Methods: We searched MEDLINE, Embase, Web of Science, Scopus, Cochrane CENTRAL, and Google Scholar for studies diagnosing CE via CD138 immunostaining, assessing microbiota with molecular techniques. Data extraction, quality assessment, and meta-analysis were performed. Results: Twenty-two studies including 4022 women were analyzed. CE was associated with reduced prevalence of Lactobacillus-dominated microbiota and increased detection of non-Lactobacillus species, particularly Streptococcus spp., Enterococcus spp., Escherichia coli, Staphylococcus spp., Ureaplasma spp., and Gardnerella vaginalis. In the meta-analysis (2947 women), Enterococcus spp. and Ureaplasma spp. were significantly more prevalent in women with CE, whereas Streptococcus spp., E. coli, Staphylococcus spp. and G. vaginalis showed non-significant trends. Only E. coli and Streptococcus spp. showed significant heterogeneity between-studies. Conclusions: CE is linked to microbial dysbiosis with reduced Lactobacillus dominance and enrichment of potentially pathogenic taxa, notably Enterococcus and Ureaplasma spp. These findings suggest that the endometrial microbiome contributes to chronic inflammation and adverse reproductive outcomes, yet heterogeneity and limited evidence call for standardized diagnostics and robust trials before clinical implementation.

Biomedicines doi: 10.3390/biomedicines14040869

Authors: Cengizhan Kurt Arif Gök

Background/Objectives: The medial meniscus is crucial for load transmission and knee stability. Meniscal tears disrupt joint biomechanics, increasing the risk of cartilage degeneration. However, few studies have quantitatively compared how different tear types affect stress and contact mechanics using finite element analysis (FEA). This study aims to analyze stress distributions for various meniscal tear types and develop a predictive model for meniscal stress behavior. This study investigates how stress distributions differ between healthy and torn medial menisci under identical loading conditions. The study examines which meniscal tear type produces the highest stress concentrations. The effects of different tear types on penetration, gap formation, pressure distribution, and sliding distance at the meniscus interface are also analysed. Materials and Methods: The FEA model of the knee joint, including femoral and tibial cartilage and the medial meniscus, was developed. Simulations were conducted for a healthy meniscus and for menisci with radial, horizontal and complex tears. Stress, penetration, gap, pressure, and sliding distance were calculated, and a mathematical model describing their relationships was established. Results: All torn menisci exhibited significantly higher stresses than the healthy meniscus (p < 0.001). Radial tears generated the highest stress concentrations (p < 0.001). Pressure was mainly influenced by meniscal geometry, while the gap remained nearly constant. Penetration increased slightly (p < 0.05). The predictive model demonstrated a strong correlation between meniscal stress and interface parameters (R2 > 0.9). In a healthy meniscus, stress distribution is homogeneous (≈26 MPa). Stress concentration increases depending on the tear type: limited in a horizontal tear (≈26.5 MPa), significant in a vertical tear (≈30.8 MPa), and highest in a radial tear (≈40.6 MPa). These results indicate that as the tear progresses, the load-bearing capacity of the meniscus decreases, and stresses concentrate at the tear edges. Conclusions: Meniscal tears, especially radial ones, substantially alter knee biomechanics and elevate tissue stress. These biomechanical insights highlight the importance of early diagnosis and targeted rehabilitation strategies to prevent further cartilage damage and osteoarthritis progression.

Biomedicines doi: 10.3390/biomedicines14040867

Authors: Jing He Ruiyun Wang Pengcheng Yang Zhuanglong Xiao Tao Bai Xiaohua Hou Lei Zhang

Objective: ZhiZhu Kuanzhong (ZZKZ) capsule, a Chinese herbal extract, is extensively employed for the clinical management of functional dyspepsia (FD) in China. This study aimed to elucidate the therapeutic efficacy and underlying mechanisms of ZZKZ on the co-morbidity of anxiety and depression of FD. Methods: The FD model was established in Sprague–Dawley rats via neonatal gastric irritation with 0.1% iodoacetamide. Subsequently, FD rats were gavaged with ZZKZ or fluoxetine. Depression-like behaviors were evaluated using the sucrose preference test (SPT) and forced swimming test (FST), while anxiety-like behaviors were assessed via light-dark box (LDB) and open field tests (OFTs). Network pharmacology and molecular docking were conducted to explore the mechanisms of ZZKZ’s action. Hippocampal levels of monoamine neurotransmitters and monoaminergic system components were evaluated by HPLC and RT-qPCR, respectively. Serum concentrations of HPA axis hormones were determined by ELISA. Results: ZZKZ administration reversed the deficits in body weight gain and food intake in FD rats. Behaviorally, ZZKZ increased sucrose consumption in SPT and prolonged swimming duration in FST, and it increased duration and entries into the central zone in OFT. According to the prediction of network pharmacology, ZZKZ treatment elevated hippocampal levels of 5-HT/NE/DA, increased expression of TPH2/TH, and decreased expression of MAOA/SERT in FD rats. Molecular docking further confirmed high-affinity binding between core ingredients of ZZKZ and TPH2/TH/MAOA/SERT. Moreover, ZZKZ administration attenuated the stress-induced elevation of serum CRH/ACTH/CORT. Conclusions: ZZKZ effectively ameliorates the disordered gut–brain interaction and mitigates anxiety-like and depression-like behaviors, which might be modulated by the hippocampal monoaminergic system and hypothalamic–pituitary–adrenal axis response.

Biomedicines doi: 10.3390/biomedicines14040868

Authors: Fangqiao Zheng Zhengyi Lan Hangrong Chen Ming Ma

Background: Osteosarcoma (OS) is an aggressive bone tumor. The lack of physiologically relevant three-dimensional models that recapitulate the native tumor microenvironment hampers drug development and mechanistic studies. The study aimed to develop bone-mimetic microspheres for the construction of an OS model. Materials and Methods: We employed droplet microfluidics to fabricate bone-mimetic microspheres (named MSHA) from a composite of gelatin methacryloyl, polyethylene glycol diacrylate, and nano-hydroxyapatite (nHA). MNNG/HOS cells were cultured on MSHA microspheres and subsequently evaluated for their bioactivity and capabilities of stemness, migration, and invasion. Results: The microfluidic platform enabled efficient and scalable production of highly uniform MSHA microspheres with controlled sizes. MNNG/HOS cells cultured on MSHA maintained high viability and spontaneously formed compact tumor spheroids after 7 days. Compared with two-dimensional cultures, cells cultured on these microsphere-based platforms exhibited enhanced migration and invasion capacities, along with increased expression of relevant biomarkers. RNA sequencing further revealed the activation of cancer-related pathways. Notably, the incorporation of nHA into microspheres amplified these malignant phenotypes, potentially through the activation of ECM–receptor interaction and calcium signaling pathways. Conclusions: The microfluidics-fabricated MSHA microspheres, as biomimetic three-dimensional culture scaffolds, offer a promising platform for applications in mechanistic studies of osteosarcoma progression and drug screening.

Biomedicines doi: 10.3390/biomedicines14040870

Authors: Panayotis K. Thanos Shtakshe Chatrath Colin Hanna Fiona Comstock John Butsch Kenneth Blum Albert Pinhasov Lucy Mastrandrea Teresa Quattrin Lesley Georger Alan Posner

Background: This study aimed to further explore the application of genetic risk assessments in 24 metabolic bariatric surgery (MBS) patients to predict weight loss outcomes three years after the procedure. Methods: Participants were assessed using the Genetic Addiction Risk Severity (GARS) test, which evaluates neurogenic polymorphisms linked to addiction and reward deficiency. Genetic and psychosocial data collected prior to surgery were analyzed in relation to post-operative weight loss measures, including weight change, body mass index (BMI), percentage of total weight loss (%TWL), and percentage of expected weight loss (%EWL). The analysis examined associations between specific genetic risk alleles, weight-related outcomes at three to four years post-surgery, and psychosocial trait scores. Results: Spearman’s correlations revealed that the DRD2 risk allele is negatively correlated with 3-year BMI (rs = −0.481, p < 0.05, 95% CI: –0.746 to –0.083). One-way ANOVA indicated that there is a significant difference in 3-year BMI (p = 0.018) between 0 and 1 DRD2 risk allele copy. There is also a significant difference in ∆weight (p = 0.022), ∆BMI (p = 0.014), and %EWL (p = 0.032) among the different SNP expression values of the MAOA risk allele. In addition, Spearman’s correlation revealed that FCQ scores are negatively correlated with ∆BMI (rs = −0.470, p < 0.05, 95% CI: −0.767, −0.005), %TWL (rs = −0.561, p < 0.05, 95% CI: −0.814, −0.129), and %EWL (rs = −0.533, p < 0.05, 95% CI: −0.800, −0.090) at 3 years post-surgery and positively correlated with 3-year weight (rs = 0.576, p < 0.05, 95% CI: 0.151, 0.821) and 3-year BMI (rs = 0.552, p < 0.05, 95% CI: 0.117, 0.810). Lastly, GARS scores are positively correlated with 3-year ∆weight (rs = 0.422, p < 0.05, 95% CI: 0.010, 0.712).

Biomedicines doi: 10.3390/biomedicines14040866

Authors: Wendy N. Phoswa Lawrence Chauke Kabelo Mokgalaboni Gaynor Balie Sidney Hanser Olive P. Khaliq

Background: Pre-eclampsia (PE) is a significant cause of maternal and perinatal morbidity and mortality globally and is characterized by impaired endothelial function and disturbances in coagulation pathways. The effects of Human Immunodeficiency Virus (HIV) on the immune and coagulation systems have been investigated during pregnancy, but there are few reports on anticoagulant factors in pregnant women who are infected with HIV and develop PE. This investigation compares plasma protein C levels in pregnant women with pre-eclampsia and those without pre-eclampsia, and compares the results based on their HIV status. Methods: A hospital-based cross-sectional study design was used for the current research, which was carried out at Charlotte Maxeke Johannesburg Academic Hospital, South Africa. A total of 83 pregnant women participated in the study and were categorized into one of four groups: normotensive HIV-negative (n = 36); normotensive HIV-positive (n = 18); pre-eclamptic HIV-negative (n = 21); and pre-eclamptic HIV-positive (n = 8). Data collected included demographic information and clinical characteristics that were abstracted from maternity records. Plasma protein C concentrations were determined by ELISA (enzyme-linked immunosorbent assay). Nonparametric statistical methods were used to compare the mean values of plasma protein C between each of the four groups, and significance was set at p < 0.05. Subgroup analyses, particularly for the pre-eclamptic HIV-positive group (n = 8), were considered exploratory due to small sample sizes. Results: As would be anticipated, both systolic and diastolic blood pressure values were significantly elevated in the pre-eclamptic group when compared to the normotensive control subjects (p < 0.0001). There were no statistically significant differences in plasma protein C concentration between the normotensive and pre-eclamptic groups, nor between the HIV-negative and HIV-positive groups. Similarly, there were no significant differences in plasma protein C concentration when comparing all four study groups (Kruskal–Wallis test p = 0.2295). Conclusions: Plasma protein C concentrations did not vary significantly according to the presence of pre-eclampsia or HIV status in this cohort. These findings suggest that protein C concentrations were not measurably altered between groups within this study population. However, due to the small sample size in key subgroups, these findings should be considered preliminary and interpreted with caution. Larger, adequately powered studies are required to further investigate potential associations between HIV infection, pre-eclampsia, and anticoagulant pathways during pregnancy.

Biomedicines doi: 10.3390/biomedicines14040865

Authors: Gianni Allais Massimo Autunno Florindo D’Onofrio Luisa Fofi Maria Gabriella Saracco Fabiola Bergandi Chiara Benedetto Francesca Silvagno Loredana Bergandi

Background: Menstrual migraine (MM), including pure menstrual migraine (PMM) and menstrually related migraine (MRM), is characterized by attacks occurring in close temporal association with menstruation and is often more severe, longer lasting, and less responsive to treatment than non-menstrual migraine. Prostaglandin-mediated inflammation and calcitonin gene-related peptide (CGRP) release play a key role in MM pathophysiology. Phycocyanin (PC) and palmitoylethanolamide (PEA) are nutraceutical compounds with anti-inflammatory, analgesic, and neuroprotective properties that may be beneficial as short-term perimenstrual prophylaxis. Objectives: To evaluate the effectiveness of an oral supplementation combining phycocyanin and palmitoylethanolamide as a short-term prophylaxis for menstrual migraine in a real-world clinical setting, a retrospective observational study without a control group was conducted in five Italian centers between May 2023 and June 2025. Methods: Clinical records of 800 women were reviewed, and 220 patients receiving perimenstrual supplementation with phycocyanin and palmitoylethanolamide were screened. Sixty-one women diagnosed with migraine without aura, according to the International Classification of Headache Disorders, met all inclusion criteria and were analyzed. Phycocyanin and palmitoylethanolamide were taken at a dosage of two capsules daily from five days before to five days after the onset of menstruation for three consecutive months. Outcomes during the perimenstrual window were compared with a three-month period without supplementation. Primary outcomes included migraine severity, frequency, and duration of the attacks; secondary outcomes included analgesic consumption and menstrual migraine-associated symptoms. Results: Among the 61 included patients, phycocyanin and palmitoylethanolamide supplementation was associated with a significant reduction in migraine severity across all monitored perimenstrual days (p < 0.0001). While the overall monthly frequency of migraine attacks did not change, the number of migraine days during the perimenstrual window significantly decreased from the first month of supplementation (p < 0.05). Moreover, migraine duration during the perimenstrual window was significantly reduced at one, two, and three months of phycocyanin and palmitoylethanolamide supplementation compared with baseline. Analgesic use and the number of days with migraine-associated symptoms (nausea, vomiting, photophobia/phonophobia) were also significantly reduced. Treatment was well tolerated. Conclusions: In this real-world retrospective study, perimenstrual supplementation with phycocyanin and palmitoylethanolamide was associated with reduced severity, duration, and perimenstrual frequency of menstrual migraine attacks, along with decreased analgesic use, suggesting a safe and potentially beneficial short-term prophylactic strategy for women with menstrual migraine.

Biomedicines doi: 10.3390/biomedicines14040864

Authors: Changxin Fan Yanyu Chen Qinghua Huang Wai Ying Ou Cancan Zhang Yanlin Sun Tianyue Wu On Yee Leung Hei Ching Iu Jiacheng Shi

Hair serves essential functions, including mechanical sensing, head protection, and body temperature regulation, while also playing a significant role in human aesthetics. However, factors such as hormonal imbalances, autoimmune disorders, infections, and psychological stress contribute to the widespread issue of hair loss, particularly among the elderly, adversely affecting self-confidence and self-esteem. Although treatments such as minoxidil, finasteride, and dutasteride have received regulatory approval, their associated side effects, such as sexual dysfunction, neuropsychiatric issues, and cardiovascular symptoms, can impede patient recovery. While follicular unit transplantation and stem cell therapy show promising outcomes, they are not suitable for all types of hair disorders. Short peptides that mimic intracellular signals and exhibit diverse biological effects have emerged as a promising approach for stimulating hair regrowth. By combining different formulations and nanosystems, the limitations of short peptides can be effectively addressed. This review systematically summarizes recent advances in peptide-based treatments for hair loss, highlighting their advantages and limitations.

Biomedicines doi: 10.3390/biomedicines14040863

Authors: Weronika Rzeska Aneta Adamiak-Godlewska

Background: Endometrial cancer (EC) is the most common gynecologic malignancy in developed countries and one of the few solid tumors with a steadily rising incidence, paralleling global trends in obesity and insulin resistance. Its strong epidemiologic association with systemic metabolic dysfunction positions EC as a uniquely accessible model for metabolically informed chemoprevention. Methods: This narrative review was conducted through a systematic search of PubMed/MEDLINE and Embase using the following terms: “endometrial cancer” AND (“insulin resistance” OR “metabolic syndrome” OR “PI3K” OR “chemoprevention” OR “bariatric surgery” OR “metformin” OR “cellular senescence”). Searches were limited to English-language publications; no date restriction was applied for foundational molecular studies, while clinical and translational evidence was reviewed from 2000 to 2025. Additional references were identified through manual review of reference lists of included articles. Results: We examine metabolic amplification as a conceptual framework in which hyperinsulinemia, inflammatory reinforcement, and redox-epigenetic modulation intensify proliferative signaling in biologically susceptible endometrial tissue, particularly within molecular subtypes enriched for PI3K pathway activation such as tumors lacking a specific molecular profile (NSMP). Bariatric surgery offers the strongest human evidence supporting the principle that durable metabolic correction can substantially reduce EC incidence. In contrast, pharmacologic interventions including metformin, anti-inflammatory agents, and nutraceutical compounds demonstrate variable or limited preventive efficacy, and short-term biomarker modulation cannot substitute for validated reduction in cancer risk. The endometrial intraepithelial neoplasia (EIN) model provides a uniquely accessible platform for biomarker-guided intervention. Conclusions: Integration of genomic subtype classification with metabolic profiling may enable precision prevention strategies in clearly defined high-risk populations. Effective chemoprevention will require molecular enrichment, confirmation of tissue-level target engagement, and clinically meaningful endpoints, while acknowledging the translational limits of pathway-directed approaches.

Biomedicines doi: 10.3390/biomedicines14040860

Authors: Shah Rezlan Shajahan Nurhidayah Hamid Blaire Okunsai Norshafarina Shari Muhammad Danial Che Ramli

Background: Alzheimer’s disease (AD) is a multifactorial neurodegenerative disorder characterized by amyloid-β aggregation, tau hyperphosphorylation, oxidative stress, and mitochondrial dysfunction. Emerging evidence indicates that the gut microbiota plays a critical role in modulating neuroinflammatory, and metabolic pathways involved in AD pathogenesis through the microbiota-gut-brain axis. Objective: This systematic review aims to comprehensively evaluate the role of the microbiota-gut-brain axis in Alzheimer’s disease, with a particular focus on its mechanistic links to oxidative stress, mitochondrial dysfunction, and amyloid pathology, as well as its therapeutic potential. Methodology: A comprehensive literature search was conducted using PubMed, Scopus, and Web of Science databases, focusing on studies evaluating gut microbiota composition, metabolomic changes, oxidative stress markers, mitochondrial activity, and therapeutic interventions in AD models and patients. Results: Altered gut microbial composition in AD is associated with increased pro-inflammatory taxa (Escherichia-Shigella, Bacteroides) and depletion of short-chain fatty acid (SCFA) producing bacteria (Faecalibacterium, Roseburia). Dysbiosis contributes to systemic inflammation, disrupted intestinal permeability, and microglial activation, leading to oxidative damage and mitochondrial impairment in neurons. Preclinical and clinical studies indicate that probiotics, prebiotics, and fecal microbiota transplantation can restore redox balance, reduce neuroinflammation, and improve cognitive outcomes. Multi-omics and AI-based models are emerging as tools for identifying microbiome-derived biomarkers for early AD detection. Conclusion: The gut microbiota-mitochondria-oxidative stress axis represents a promising therapeutic target in Alzheimer’s disease. Future research should focus on longitudinal human studies, standardized microbial profiling, and personalized microbiome-based interventions to translate these mechanistic insights into clinical benefit.

Biomedicines doi: 10.3390/biomedicines14040862

Authors: Mehmet İncebıyık Adalet Göçmen

Objective: To identify clinical and neuroimaging predictors of atypical Posterior Reversible Encephalopathy Syndrome (PRES) in eclampsia and evaluate the role of multi-regional cerebral involvement (neuroimaging burden). Methods: This retrospective cohort study included 266 patients with eclampsia and radiologically confirmed PRES (2018–2025). Patients were classified as typical (n = 234, 88.0%) or atypical (n = 32, 12.0%). A two-stage multivariable logistic regression was performed to identify independent predictors, sequentially incorporating clinical and neuroimaging variables. Results: Peak systolic blood pressure was significantly higher in atypical vs. typical groups (191.6 ± 20.4 vs. 172.4 ± 18.5 mmHg, p < 0.001). Furthermore, atypical cases exhibited a significantly higher systemic inflammatory burden, characterized by markedly elevated Systemic Immune-Inflammation Index (SII) and CRP levels (p < 0.001). Atypical cases exhibited a markedly greater neuroimaging burden, with a higher mean number of involved brain regions (4.4 ± 1.2 vs. 2.1 ± 0.6, p < 0.001). In Model 1 (clinical variables only), systolic blood pressure was a strong predictor of atypicality (OR: 1.24 per 10 mmHg increase, 95% CI: 1.12–1.38, p < 0.001). After incorporating neuroimaging features in Model 2, the total number of involved brain regions emerged as the strongest independent predictor (OR: 2.08, 95% CI: 1.52–2.85, p < 0.001), while the independent effect of blood pressure was attenuated. Conclusions: Atypical PRES in eclampsia reflects extensive, high-burden cerebral vasogenic edema rather than a distinct radiological subtype. While hypertension initiates the process, the total regional burden determines the atypical signature. This burden-focused perspective improves risk stratification and diagnostic vigilance in high-risk obstetrics.

Biomedicines doi: 10.3390/biomedicines14040861

Authors: Antonio Bueno-Nava Diana-Karina Díaz-Hernández Rogelio Paniagua-Pérez Paul Carrillo-Mora José-Antonio Martínez-Cortez Claudia Hernández-Arenas Saúl-Renán León-Hernández Adriana Olmos-Hernández Antonio Verduzco-Mendoza Alberto Avila-Luna Arturo Gálvez-Rosas

Introduction: The histaminergic pathway has been implicated in Parkinson’s disease (PD). Histamine is metabolized by histamine N-methyltransferase (HNMT), and the gene encoding this enzyme has a C314T polymorphism, in which cytosine is replaced by thymine. This results in reduced enzymatic activity. Objective: To analyze the C314T polymorphism of the HNMT gene in Mexican patients with idiopathic PD. Materials and Methods: In this study, peripheral blood samples were collected from patients with PD and healthy controls for genomic DNA extraction. HNMT genotyping was performed using the restriction fragment length polymorphism (RFLP) technique. Quantitative variables were compared using Student’s t test, and categorical variables were compared using Pearson’s χ2 test. The risk of PD was estimated using odds ratios (ORs) and 95% confidence intervals (CIs). Results: According to the results of the bivariate analysis, compared with the controls, the patients were significantly older (p = 0.001) and had a higher incidence of hypertension (p = 0.020). HNMT RFLP analysis suggested an association between the C allele and PD development, with an OR (95% CI) of 7.424 (0.866–63.646). In contrast, the T allele appeared to confer a protective effect, with an OR of 0.134. In the age-adjusted Mantel—Haenszel stratified analysis of the HNMT C314T polymorphism, the C allele was identified as a risk factor for PD development in this small cohort, with an OR (95% CI) of 12.0 (0.8–160.4; p = 0.041). Conclusions: Advanced age, hypertension, and the C allele of the HNMT gene were associated with an increased risk of PD, whereas the T allele appeared to be associated with a protective role.

Biomedicines doi: 10.3390/biomedicines14040859

Authors: Yujia Lin Panpan Lu Qiang Ding Mei Liu

The pathogenesis of inflammatory bowel disease (IBD) is driven by an interplay among intestinal dysbiosis and aberrant mucosal immune responses. This review centers on the microbiota as a pivotal pathogenic hub, systematically dissecting how three hallmark features of dysbiosis—reduced microbial alpha diversity, depletion of immunomodulatory commensals, and expansion of pro-inflammatory pathobionts—collectively compromise epithelial barrier function, promote bacterial translocation, and sustain chronic mucosal inflammation. We further integrate emerging evidence implicating bidirectional gut-brain axis communication in amplifying both peripheral inflammation and central nervous system (CNS)-mediated behavioral comorbidities. Building on this mechanistic framework, we critically evaluate next-generation microbiota-targeted interventions: standardized fecal microbiota transplantation (FMT), rationally designed live biotherapeutic products (LBPs), precision phage cocktails targeting defined pathobionts, and microbiome-informed dietary strategies. Collectively, these approaches represent a paradigm shift—from broad-spectrum immunosuppression toward mechanism-guided, ecosystem-level modulation—thereby advancing the goal of precision medicine in IBD.

Biomedicines doi: 10.3390/biomedicines14040858

Authors: Xin-Fang Leong

Cardiovascular disease continues to impose a substantial global health burden and arises from interconnected pathological processes, including oxidative injury, inflammatory signaling, endothelial dysfunction, metabolic imbalance, and progressive cardiac and vascular structural remodeling. Growing interest has therefore emerged in naturally derived compounds capable of influencing multiple disease pathways simultaneously. Pterostilbene, a dimethoxylated stilbene structurally related to resveratrol, has gained attention due to its enhanced lipophilicity and improved bioavailability. Recent experimental studies have investigated the cardiovascular effects of pterostilbene in both cellular systems and animal models. Evidence from in vitro studies indicates that this compound modulates key regulatory networks involved in cellular energy metabolism, redox homeostasis, endothelial signaling, and stress-associated cardiomyocyte injury. These actions involve pathways linked to 5′ adenosine monophosphate-activated protein kinase and sirtuin-1 signaling, nitric oxide regulation, antioxidant responses, and ferroptosis-related mechanisms. Findings from in vivo investigations further demonstrate protective effects across multiple cardiovascular disease models, including pulmonary hypertension, pressure overload-associated cardiac remodeling, ischemic myocardial injury, toxin-induced cardiotoxicity, and metabolic or atherosclerotic vascular dysfunction. Improvements in functional, structural, and biochemical parameters have been reported in these experimental settings. Overall, current preclinical evidence suggests that pterostilbene may act as a multifunctional modulator of key processes involved in cardiovascular pathology. Although clinical evidence remains limited, the convergence of mechanistic and experimental findings highlights its potential as a multi-target cardiometabolic therapeutic candidate and provides a foundation for future translational and clinical investigation.

Biomedicines doi: 10.3390/biomedicines14040857

Authors: Anton Kolarov Irina Chakarova Valentina Hadzhinesheva Venera Nikolova Stefka Delimitreva Maya Markova Ralitsa Zhivkova

Background/Objectives: Osteoarthritis has been increasingly described as associated with systemic inflammation, raising the question of how it would affect fertility in young women with or without reproductive hormone administration. We studied oogenesis in mice with collagenase-induced osteoarthritis (CIOA) as a model system with fewer ethical limitations after estradiol (E2) or follicle-stimulating hormone (FSH) treatment. Methods: Oocytes have been isolated from mice subjected to various treatment regimens. The meiotic spindle, the chromatin, and the actin cap were fluorescently labeled and analyzed. Results: In addition to reduced maturation rates, specific oocyte abnormalities were registered when CIOA, FSH, or E2 were applied in isolation. Combined treatments showed that the spindle, chromatin, and actin cytoskeleton parameters were differently affected in oocytes from groups with CIOA treated by estradiol and those treated with FSH. Enlarged spindles, ooplasmic tubulin asters, aligned metaphases, and predominantly normal actin caps, often with an actin halo, were typical for groups with CIOA combined with estradiol. The groups with CIOA and FSH had slightly enlarged spindles, unaligned metaphases with degenerated chromatin surrounded by a cloud of depolymerized tubulin, and small actin caps. Conclusions: Our results show that experimental osteoarthritis with or without exogenous reproductive hormones negatively affects oogenesis, presumably due to systemic inflammatory factors making the ovarian microenvironment less capable of supporting oocyte maturation. Estradiol supplementation does not benefit oogenesis. FSH treatment induced cytoskeletal and chromatin abnormalities that presumably disturb the fertilization and development potential of affected oocytes. These data can have implications for assisted reproduction in cases of patients with osteoarthritis.

Biomedicines doi: 10.3390/biomedicines14040855

Authors: Anja-Maria Ladek Leonie Priebe Thomas Harrer Ellen Harrer Georg Michelson Thomas S. Knauer Diogo X. Dias-Nunes Christian Y. Mardin Antonio Bergua Bettina Hohberger

Background: Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a debilitating disorder, characterized by symptoms such as post-exertional malaise (PEM) and cognitive impairments. This study assessed reaction time (RT) metrics in three-dimensional (3D) visual tasks with the aim of objectively quantifying the cognitive impairments in ME/CFS patients compared to controls. Methods: A total of 120 participants (60 ME/CFS patients and 60 controls) were recruited at the Department of Ophthalmology, Universität of Erlangen-Nürnberg. RT was assessed using a virtual reality–oculomotor test system, presenting 3D stimuli at three disparity levels (275″, 550″, and 1100″) within three gaming repetitions (R1, R2, and R3). Mixed-effects models were used to evaluate group differences, with age and gender as covariates. Pairwise contrasts were calculated to assess changes across repetitions. Fatigue self-assessments were recorded by validated questionnaires, (FACIT Fatigue Scale, Chalder Fatigue Scale, Bell Score and Health Assessment Questionnaire), and their correlation with RT metrics was portrayed using a Spearman correlation matrix. Results: Estimated means (EM-means) for RT were significantly prolonged in ME/CFS patients compared to controls at disparity 275″ (1969 ms vs. 1384 ms; p = 0.0001), 550″ (1409 vs. 1071 ms; p = 0.0012) and 1100″ (1126 ms vs. 891 ms; p = 0.00223). Age was a significant covariate (p < 0.001), while gender showed no effect. Both groups demonstrated improvements in RT over repetitions; however, ME/CFS patients showed a significantly lower improvement compared to controls, reaching significance in R3 (p = 0.0042). RT metrics did not correlate with patients’ self-assessment scores. Conclusions: ME/CFS patients showed consistently slower RTs compared to controls, particularly in later, easier gaming repetitions, potentially reflecting the impact of fatigue.

Biomedicines doi: 10.3390/biomedicines14040856

Authors: Didik Setyo Heriyanto Fahrul Nurkolis Jinwon Choi Sohyun Park Min Choi Raymond Rubianto Tjandrawinata Amama Rani Moon Nyeo Park Min-Jin Kwak Bum Sang Shim Bonglee Kim

Lung cancer remains the leading cause of cancer-related mortality worldwide, with non-small cell lung cancer accounting for the majority of cases and exhibiting persistent challenges related to therapy resistance and metastatic progression. Increasing evidence indicates that dysregulated G protein-coupled receptor signaling and ion channel activity function cooperatively as master regulators of tumor cell proliferation, migration, survival, and therapeutic response. Cannabinoids, including phytocannabinoids such as delta-9-tetrahydrocannabinol and cannabidiol, as well as endogenous endocannabinoids, are uniquely positioned to modulate both G protein-coupled receptors and ion channels, thereby influencing key oncogenic signaling networks. This review synthesizes current knowledge on the role of major ion channel families, including transient receptor potential channels, potassium channels, and sodium channels, and principal G protein-coupled receptor pathways involved in lung cancer progression. We further discuss how cannabinoids reprogram these interconnected signaling systems through canonical cannabinoid receptors, non-classical targets such as G protein-coupled receptor 55 and adenosine receptors, and direct modulation of ion channel activity. Special attention is given to G protein-coupled receptor–ion channel coupling within membrane microdomains and to the capacity of cannabinoids to act as biased ligands, redirecting downstream pathways, such as the phosphoinositide 3-kinase–protein kinase B–mechanistic target of rapamycin and epidermal growth factor receptor signaling, toward apoptosis and reduced metastatic potential. Emerging strategies, including cannabinoid-based combination therapies, selective receptor biasing, and targeted delivery systems, are also highlighted. Altogether, cannabinoid-driven rewiring of G protein-coupled receptor and ion channel signaling represents a promising mechanistic framework for developing innovative therapeutic approaches against lung cancer.

Biomedicines doi: 10.3390/biomedicines14040854

Authors: John A. Dangerfield Christoph Metzner

Regenerative medicine is becoming more widely integrated with longevity-oriented and preventive care as populations age and chronic degenerative diseases burden healthcare systems. Mesenchymal stem cell (MSC) therapies have progressed from experimental interventions to approved products, yet scalability, safety, cost, and regulatory complexity constrain widespread implementation in medical wellness contexts. The predominant therapeutic effects of MSCs are mediated via paracrine mechanisms, leading to cell-free approaches based on the MSC secretome—a complex mixture of bioactive factors including all types of biomolecules and assemblies thereof, such as exosomes. These acellular products offer compelling advantages: multiple batches from single-donor sources, standardized dosing, reduced allogeneic cell risks, and shorter outpatient-compatible administration. Preclinical and clinical data indicate that secretome-based products exert potent regenerative effects in osteoarthritis, chronic wounds, stroke, traumatic brain injury, and neurodegenerative diseases. This review examines the evolution from cell-based to cell-free regenerative strategies, focusing on human umbilical cord Wharton’s jelly MSC secretome for precision longevity medicine. It compares MSC therapies with secretome- and exosome-based formulations across mechanistic, manufacturing, safety, practical and regulatory dimensions. Regional perspectives highlight Southeast Asia, and especially Thailand, as an emerging regenerative-longevity hub. Finally, it outlines the preventive patient journey integrating cell-free interventions within multi-modal programs aimed at extending healthspan.

Biomedicines doi: 10.3390/biomedicines14040851

Authors: Yafeng Ma Ya-Wen Chiang Therese M. Becker Jon Hyett

Preeclampsia (PE), pregnancy-associated high blood pressure linked to organ damage, affects 3–8% of all pregnancies and results worldwide in 70,000 maternal and 500,000 perinatal deaths each year. Untreated PE may progress to eclampsia with long-term health implications for both mother and child. Non-invasive prenatal diagnosis or screening applies cell-free DNA approaches and offers a less invasive and more economical method for early diagnosis and prediction of various pregnancy complications. Recently, cell-free assays, particularly blood-based cell-free DNA and RNA analysis, have shown great potential in early PE prediction and diagnosis. Here, we provide an updated review of the current understanding and discoveries of PE, focusing on recent publications (1 January 2019–30 December 2025) of liquid biopsy-derived circulating fetal cells (circulating trophoblasts and fetal nucleated red blood cells), cell-free DNA, cell-free RNA and small extracellular vesicles (i.e., exosomes). We aim to discuss the conceptual framework and technical evolution of liquid biopsy applications in preeclampsia pathogenesis, prediction and diagnosis. Progressing novel screening and diagnostic molecular biomarkers have high potential to facilitate early detection for patients at risk of PE. Liquid biopsy-based screening strategies may aid in providing timely intervention and treatment.

Biomedicines doi: 10.3390/biomedicines14040853

Authors: Stephen Shei-Dei Yang Kuan-Yu Chen Earl Fu Hsi-Hsien Chang Kuo-Feng Huang

Background/Objectives: The renin–angiotensin–aldosterone (RAA) system is a key regulator of cardiovascular homeostasis. Recent evidence suggests that Angiotensin II (Ang II) can trigger ferroptosis, an iron-dependent form of cell death. We previously demonstrated that periodontitis induces neurovascular dysfunction, and our preliminary observations indicate that this oral inflammatory model is associated with elevated blood pressure. However, the mechanism by which Ang II impaired nitrergic vasodilation and triggered ferroptosis in cerebral arteries remains unclear. This study investigates the functional effects of electrical and chemical nerve stimulation in adult spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY). Methods: Endothelium-denuded basilar arterial (BA) rings from SHRs and WKYs were used to assess the impact of Ang II on neurogenic relaxation via wire myography. Results: Vascular relaxation responses to nicotine and transmural nerve stimulation (TNS) were significantly diminished in SHRs compared to WKYs. This impairment was reversed by both acute preincubation and chronic treatment with losartan (an AT1 receptor antagonist). In WKY BAs, exogenous Ang II pretreatment inhibited relaxation responses to nicotine, TNS, and isoproterenol. Importantly, this inhibition was effectively reversed by marimastat (MMP inhibitor), catalase (antioxidant), and ferrostatin-1 (ferroptosis inhibitor). Conclusions: Our findings indicate that Ang II induces functional alterations in neurovascular signaling patterns by triggering ferroptosis within nerve terminals. This process leads to a functional imbalance between sympathetic and parasympathetic influences, ultimately impairing neurogenic nitrergic dilation in the BAs of SHRs. These results suggest that targeting Ang II-induced ferroptosis may alleviate the neuroinflammation and cognitive decline associated with hypertension-related cerebrovascular dysfunction.

Biomedicines doi: 10.3390/biomedicines14040852

Authors: Tihana Nađ Martina Kos Ana Stupin Ines Drenjančević Nikolina Kolobarić Zrinka Mihaljević Petar Šušnjara Mia Damašek Darjan Kardum Ivana Jukić

Objective: Vascular function serves as an early indicator of cardiovascular (CV) risk. The intake of n-3 polyunsaturated fatty acids (PUFAs) has been reported to improve arterial properties and reduce CV risk, but evidence in healthy individuals remains limited. This study investigated the effects of consuming n-3 PUFAs-enriched chicken meat on vascular reactivity at both microvascular and macrovascular levels in healthy young adults. Materials and Methods: In this placebo-controlled, double-blind, randomized interventional trial (ClinicalTrials.gov: NCT05725486), 39 participants (aged 20–26 years) were assigned to either the Control group (n = 20; approximately 118 mg n-3 PUFAs/day) or the n-3 PUFA group (n = 19; approximately 1500 mg n-3 PUFAs/day) for three weeks. Microvascular reactivity was assessed via post-occlusive reactive hyperemia (PORH), acetylcholine-induced dilation (AChID), local thermal hyperemia (LTH), and sodium nitroprusside-induced (SNPID) responses. Macrovascular reactivity was measured by brachial artery flow-mediated dilation (FMD) and nitroglycerine-mediated dilation (NTG-MD). Body composition and blood pressure (BP) were recorded before and after the intervention. Results: Both microvascular (PORH, AChID, and LTH) and macrovascular (FMD) endothelium-dependent vasodilation increased in the n-3 PUFAs group following the dietary protocol compared to the Control group. Conversely, the three-week dietary intervention did not influence endothelium-independent dilation in either the microvasculature (SNPID) or macrovasculature (NTG-MD) within the groups compared to baseline, nor were any differences observed between the groups. No significant changes were noted in BP or body composition after either diet. Conclusions: In healthy young adults, consuming the n-3 PUFAs-enriched chicken meat for three weeks improved endothelium-dependent vasodilation in both micro- and macrocirculation, without affecting endothelium-independent responses. These findings suggest that dietary n-3 PUFA intake may provide vascular benefits even in healthy, disease-free individuals at rest.

Biomedicines doi: 10.3390/biomedicines14040850

Authors: Qilei Chen Yuhang Ma Yingyi Liu Xiaojie Wang Guanhua Huang Yizhao Yang Joshua Ka-Shun Ko Hubiao Chen

Immune-mediated inflammatory diseases, such as rheumatoid arthritis, multiple sclerosis, and systemic lupus erythematosus, impose a severe and growing global health burden, where current therapies are limited by poor specificity and significant side effects. The peptidylarginine deiminase (PAD)/citrullination axis, in which protein citrullination catalyzed by PADs drives autoantigen generation and sustains inflammation, has emerged as a critical therapeutic target. This review outlines a comprehensive strategy for targeting this axis using natural products. We first detail the established role of natural compounds as direct PAD inhibitors, covering their chemical diversity, inhibitory mechanisms, and therapeutic applications in disease models. Subsequently, the discussion extends to their broader, indirect modulatory functions, highlighting how these compounds can suppress pathogenic citrullination by regulating upstream processes like NETosis and inflammatory signaling. Furthermore, the review introduces the innovative substrate-centric intervention strategy, which represents a paradigm shift toward shielding key arginine residues on autoantigens, thereby preventing the formation of immunogenic neoepitopes. The translational challenges and future directions for each of these avenues are outlined, addressing persistent obstacles including achieving isoform selectivity and biomarker validation. By integrating these multifaceted strategies, from direct inhibition and indirect modulation to substrate protection, this work provides a strategic roadmap for advancing the next generation of more precise, effective, and safe anti-citrullination therapies, ultimately moving beyond conventional enzyme inhibition toward targeted immunomodulation in immune-mediated inflammatory diseases.

Biomedicines doi: 10.3390/biomedicines14040849

Authors: María F. Naranjo-Ortíz Luz Parada-Rubio José Fuentes-Montoya Jean Carlos Villamil Poveda Francy Elaine Torres-Suarez Heidy-C. Martínez-Díaz Laura Daniela Ardila Ortiz Juliana Velosa-Porras Lorenza Jaramillo Jorge Andrés Castillo Jairo Jaime Nelly S. Roa Adriana P. Corredor-Figueroa

Background: Infections caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have generated major public health concerns worldwide. Young adults represent a critical group for viral transmission due to their high proportion of asymptomatic infections. Objective: To characterize the dynamics of SARS-CoV-2-specific antibodies in individuals aged 20–29 years from Bogotá, Colombia, across two longitudinal phases. Methods: Phase I assessed seroprevalence, seroconversion, spatial clustering, symptoms associated with seropositivity and antibody kinetics following natural infection. Phase II evaluated vaccine-induced antibodies, immune memory, and neutralizing capacity. Analyses included Functional Principal Component Analysis, survival analysis, clustering, and predictive modeling. Results: In Phase I, a seroprevalence of 15.59% (17/109 participants enrolled) was observed, while seroconversion among those who completed all six sampling points was 30.18% (16/53), with clusters of positive cases in different areas of Bogotá. The symptoms most associated with seropositivity included mucus hypersecretion, fever, and respiratory difficulty. Antibody responses were heterogeneous: naturally infected individuals generally showed high titers during the first 1–2 months, remaining detectable up to 4 months. The reduction in dimensionality suggested dominant humoral patterns, and clustering revealed two immune profiles differing in the risk of seroconversion. Predictive modeling indicated diverse antibody trajectories over 12 months. In Phase II (2024), three long-term immune memory clusters (low, medium, high) were observed; post-vaccination IgG titers were observed, although in most cases they lacked neutralizing activity. Conclusions: This longitudinal exploratory observational study provides an initial characterization of antibody dynamics in young adults, suggesting their potential epidemiological relevance and offering preliminary insights into post-infection and post-vaccination immunity.

Biomedicines doi: 10.3390/biomedicines14040848

Authors: Monica Denisa Elena Popescu Costel-Valentin Manda Octavian Croitoru Daniela-Maria Calucică Johny Neamțu Andrei Biță Amelia Maria Găman Simona-Daniela Neamțu

Background/Objectives: Epirubicin, Olaparib, and Ribociclib are widely used anticancer agents whose serum concentrations exhibit significant inter-individual variability, supporting the need for reliable and robust analytical methods suitable for pharmacokinetic evaluation and therapeutic exposure assessment. Variations in metabolism, drug–drug interactions, organ function, and treatment regimens may substantially influence systemic exposure, highlighting the importance of accurate quantification in clinical practice. This study describes the development and validation of a solid-phase extraction–liquid chromatography–mass spectrometry (SPE–LC–MS) method for the simultaneous quantification of these drugs in human serum. Methods: Sample preparation was performed using Oasis PRiME HLB® cartridges to ensure efficient clean-up, optimal recovery, and reduced matrix effects. Chromatographic separation was achieved using gradient elution with 0.1% formic acid and acetonitrile on a reversed-phase column, followed by single-quadrupole mass spectrometric (QDa) detection in the selected ion recording mode. The total run time was 13 min, enabling high-throughput analysis. Results: The method demonstrated good linearity (r > 0.997) over the tested concentration ranges, along with adequate selectivity, precision, accuracy, recovery, and stability, fulfilling the ICH M10 guideline validation criteria. No significant carry-over or interference from endogenous compounds was observed. Conclusions: Application to patient samples confirmed reliable performance in real clinical matrices and consistent quantification across different concentration levels. The proposed approach provides a potentially more accessible alternative in laboratories already equipped with LC-MS systems compared to LC-MS/MS platforms and can be applied in pharmacokinetic studies, representing a proof-of-concept for exposure assessment in oncology.

Biomedicines doi: 10.3390/biomedicines14040847

Authors: Sanja Dimic-Janjic Ivana Buha Jelena Cvejic Nikola Kostadinovic Slavko Stamenic Anka Postic Ana Ratkovic Kristina Stosic-Markovic Ivana Sekulovic-Radovanovic Marija Vukoja Nikola Trboljevac Lidija Isovic Ruza Stevic Nikola Colic Katarina Lukic Spasoje Popevic Natasa Djurdjevic Milan Savic Nikola Subotic Mihailo Stjepanovic

Background: Low-dose computed tomography (LDCT) lung cancer screening (LCS) frequently identifies emphysema in high-risk smokers. However, the extent to which CT-detected emphysema reflects underlying physiological impairment remains uncertain. We evaluated whether spirometry can detect functional abnormalities in this population beyond structural imaging findings. Methods: This cross-sectional study included 323 individuals with LDCT- detected emphysema and no lung cancer or prior chronic respiratory diseases within a screening cohort (n = 3076). Participants underwent pre-bronchodilator spirometry and symptom assessments (COPD Assessment test (CAT) and Modified Medical Research Council (mMRC) Dyspnea Scale). Pre-bronchodilator airflow limitation was defined as forced expiratory volume in one second to forced vital capacity ratio (FEV1/FVC) < 0.70. Small airways dysfunction was defined by ≥2 reduced mid-expiratory flow parameters (<60% predicted). Flow–volume curve morphology was assessed qualitatively. Results: Pre-bronchodilator airflow limitation was observed in 45.2% of participants, predominantly mild. Small-airway dysfunction was present in 52%, and an abnormal flow–volume curve morphology in 67.5%. Notably, functional abnormalities were frequently observed despite preserved FEV1. Symptom burden was low, with only 7.7% of participants reporting clinically significant symptoms. Functional impairments often overlapped and were common in minimally symptomatic individuals. Conclusions: In a lung cancer screening (LCS) cohort with CT-detected emphysema, functional abnormalities are frequently observed, including in individuals with preserved FEV1 and minimal symptoms. Spirometry provides additional physiological insight beyond structural imaging; however, these findings are descriptive and should not be interpreted as diagnostic of COPD. Further studies are needed to determine their clinical relevance.

Biomedicines doi: 10.3390/biomedicines14040846

Authors: Shasha Mei Hua Chen Jiezhi Dai

Background: Diabetic foot ulcers (DFUs) represent a severe and costly complication of diabetes mellitus. This meta-analysis aims to compare the efficacy of ultrasound-assisted wound debridement (UAWD) and conventional debridement in promoting wound healing in patients with DFUs. Methods: A systematic literature search was conducted using PubMed, EMBASE, BIOSIS, Web of Science, and the Cochrane Library from inception to 31 October 2025. Randomized controlled trials (RCTs) that compared UAWD with a placebo or standard wound care in patients with DFUs were included. Primary outcomes were the healing rate, time to complete healing, and reduction in wound area. Results were expressed as odds ratios (ORs) or mean differences (MDs) with 95% confidence intervals (CIs). This study was registered on the PROSPERO platform (CRD420251229633). Results: Ten RCTs that involved 386 patients were included. The meta-analysis showed that the treatment group had a significantly higher complete wound healing rate compared with the control group (OR: 2.92; 95% CI: 1.82 to 4.70; p = 0.75; I2 = 0%). The rate of wound area reduction was also significantly greater in the treatment group (MD: 21.29%; 95% CI: 3.03 to 39.56; p = 0.003; I2 = 75%). Furthermore, the time to complete healing was significantly shorter in the treatment group (MD: −4.84 weeks; 95% CI: −8.65 to −1.03; I2 = 61%, p = 0.05). Conclusions: UAWD appears to be more effective than conventional debridement alone in improving healing rates and accelerating wound closure in diabetic foot ulcers. However, safety data were inadequately reported across most included studies, with adverse events poorly characterized. Future large-scale RCTs should prioritize rigorous adverse event reporting to establish both the efficacy and safety profile of this intervention.

Biomedicines doi: 10.3390/biomedicines14040845

Authors: Zhouxia Xiang Wenqian Wei Shunian Guo Hanyu Meng Shu Rong

Objectives: To investigate the effects of Roxadustat and recombinant human erythropoietin (rHuEPO) on glycemic control and glycated hemoglobin (HbA1c) in non-dialysis type 2 diabetic kidney disease (DKD) patients with anemia. Methods: This retrospective study enrolled 449 patients, who were divided into three groups—the rHuEPO group (n = 252), the Roxadustat group (n = 102), and the switch group (n = 95)—in which patients were converted from rHuEPO to Roxadustat. All treatments lasted for more than three months. Changes in HbA1c and other indicators within groups as well as differences among groups were evaluated. Results: In the rHuEPO group, HbA1c levels decreased from 7.08 ± 1.19 to 6.41 ± 0.60 (p < 0.001), and they returned to baseline levels by 6–12 months (p > 0.05). In the Roxadustat group, HbA1c fluctuated but none of the differences reached statistical significance (p > 0.05). In the switch group, HbA1c decreased during rHuEPO treatment (p < 0.05) and returned to baseline after switching to Roxadustat (p > 0.05). No significant changes in blood glucose levels were observed in any group after treatment (p > 0.05). Multivariate linear regression analysis showed that changes in iron metabolism parameters, erythrocyte parameters, inflammatory markers, and glucose-lowering or lipid-lowering regimens had no significant effect on the change in HbA1c in the Roxadustat group (F = 0.834, p = 0.620), while the multivariate model of rHuEPO group also lacked statistical significance (F = 1.142, p = 0.170). After treatment, all three groups showed improvements in anemia, iron metabolism, renal function, inflammatory markers, and lipid profiles compared with baseline (p < 0.05). Additionally, further improvements in these parameters were observed after the transition from rHuEPO to Roxadustat (p < 0.05). Compared with rHuEPO group, the Roxadustat group exhibited significantly greater increases in hemoglobin, red blood cell count, total iron-binding capacity, transferrin, and serum iron (p < 0.05). Conclusions: In non-dialysis DKD patients with anemia, rHuEPO can significantly decrease HbA1c values, while Roxadustat does not. Roxadustat offers advantages over rHuEPO in terms of efficacy and assessment of glycemic control.

Biomedicines doi: 10.3390/biomedicines14040844

Authors: Dan Faganeli Metoda Lipnik-Stangelj

Cannabinoid receptors occupy strategic control nodes within motor circuitry, making them potential targets for modulating different motor manifestations. They are positioned both within basal ganglia circuits that regulate movement and within spinal circuits that control skeletal muscle tone. Consequently, cannabinoids have been studied across diverse motor disorders, most notably in movement disorders and tone disorders, particularly those resulting in spasticity. Because motor control spans multiple anatomically and functionally distinct levels, relating cannabinoid signaling to effects on motor function is not straightforward. Limited understanding of cannabinoid receptor distribution has led to cannabinoids being tested even in disorders where receptor localization would predict little or no benefit. Mapping receptor distribution within individual motor circuits and integrating them with their pharmacological effects can help anticipate how cannabinoid signaling shapes motor output. Combined with characteristic motor manifestations, one can identify motor disorders in which cannabinoids may have therapeutic value. In this review, we integrate existing evidence to place cannabinoid receptors within key motor pathways, ranging from basal ganglia circuits controlling movement to peripheral mechanisms governing muscle tone. We consider both cannabinoid 1 receptor (CB1R) and cannabinoid 2 receptor (CB2R), with CB2R gaining attention only recently for its potential relevance within the central nervous system. Building on this framework, we infer how cannabinoids acting at these sites may modulate motor control, and consequently, influence motor manifestations across major motor disorders. Finally, we examine how these distribution-based expectations align with available clinical observations.

Biomedicines doi: 10.3390/biomedicines14040843

Authors: Gabriela Calin Mihnea Costescu Marcela Nour Camer Salim Nicu Lungu Alina Stefanache Roman Rusnac Elena Costescu Mihai Cozmin Petruta Moraru Alina Mitocaru Tatiana Iov Letiția Duceac

Fractures are becoming a bigger and bigger global health problem, with an estimated 178 million new cases each year and 455 million people living with disabilities caused by fractures. Donor site morbidity, the risk of immune rejection, and limited functional integration all make current grafting techniques less effective. Biomaterials that come from nature, like collagen, gelatin, chitosan, alginate, hyaluronic acid (HA), and silk fibroin, have become promising scaffolds because they are bioactive, mimic the extracellular matrix (ECM), and can be broken down by enzymes. Crosslinking and composite reinforcement can greatly change how well they work. For example, collagen scaffolds that are highly crosslinked with glutaraldehyde keep up to 51.9% of their tensile strength after being exposed to enzymes, while non-crosslinked scaffolds only keep 12% of their strength. Chitosan–hydroxyapatite matrices, on the other hand, can reach compressive strengths of 2–12 MPa, which is close to the strength of cancellous bone. Additive manufacturing and 4D printing allow for precise control of structures and the ability to change their shape over time, which helps with vascularization and mechanical adaptation. Injectable and in situ-forming hydrogels show clinically important results, such as filling 85% of osteochondral defects in rabbits, improving left ventricular ejection fraction by up to 9% in large-animal cardiac models, and speeding up healing by 25–40% in chronic wounds. Even with these improvements, it is still hard to get batch consistency, a standardized way to test mechanical properties, and production that meets GMP (Good Manufacturing Practices) standards and can be scaled up.

Biomedicines doi: 10.3390/biomedicines14040842

Authors: Ioana Maria Suciu Călin Muntean Laura Gaiță Teodora Mateoc-Sîrb Daliborca Cristina Vlad Bogdan Timar Dan Gaiță

Background/Objectives: Sodium–glucose cotransporter-2 (SGLT2) inhibitors have demonstrated cardiovascular benefits beyond glycemic control, yet the specific biological pathways potentially linking SGLT2 inhibitor exposure to cardiovascular outcomes after acute myocardial infarction (AMI) remain incompletely characterized. Two biologically plausible pathways, serum uric acid (SUA) reduction and renal functional preservation, have been proposed, but not directly compared in a unified analytical framework. This study aimed to explore whether associations between SGLT2 inhibitor exposure and recurrent post-AMI outcomes may be more strongly linked to SUA reduction and to renal functional changes, using a hypothesis-generating causal mediation analysis. Methods: This retrospective observational cohort study included 142 consecutive patients hospitalized for AMI who underwent percutaneous coronary intervention (PCI) during the index hospitalization, reflecting standard-of-care management for AMI in this tertiary center. Patients were categorized by SGLT2 inhibitor exposure (n = 57) vs. controls (n = 85). Both diabetic (47.2%) and non-diabetic (52.8%) patients were included. The primary endpoint was change in SUA (ΔUA); the secondary endpoint was myocardial infarction (MI) recurrence. Causal mediation analysis with nonparametric bootstrap simulation tested both mechanistic pathways. Results: SGLT2 inhibitor therapy was associated with significant SUA reduction (ΔUA = −0.99 mg/dL vs. +0.56 mg/dL in controls; p < 0.001), consistent across diabetic and non-diabetic subgroups and independent of AMI recurrence. Each 1 mg/dL decrease in SUA was associated with lower odds of recurrent MI in the initial model (β = −0.25; p = 0.041). However, after incorporation of renal functional change, the uric acid-mediated pathway lost significance (ACME p = 0.462), whereas the renal-mediated pathway remained significant (ACME p = 0.038). Serum creatinine change emerged as the strongest independent predictor of MI recurrence (β = 2.22; p = 0.015). Conclusions: The findings are more consistent with a renal-mediated pathway than with an independent uric acid-mediated pathway in explaining the observed associations between SGLT2 inhibitor exposure and recurrent post-AMI outcomes. These hypothesis-generating results from a retrospective design warrant prospective validation.

Biomedicines doi: 10.3390/biomedicines14040841

Authors: Pamela Soto-Santillan Andres Jacobo-Ruvalcaba Michael Eduard Wasung-de Lay Oscar Orihuela-Rodriguez

Heart failure (HF) affects approximately 64 million people globally. HF often coexists with chronic kidney disease. HF may affect the heart during diastolic filling, systolic ejection, or both. Conventionally, HF is categorized by left ventricular ejection fraction (LVEF). One of the leading causes of death in chronic kidney disease (CKD) patients of cardiovascular origin increase hospitalizations and worsen quality of life by causing fluid and electrolyte overload. As kidney function declines, increases risk of development of HF in CKD, with a negative impact and worse prognosis in these patients. This narrative review provides healthcare professionals—including nephrologists, car-diologists, internists, and general practitioners—with evidence-based strategies to iden-tify and manage this complex comorbidity, aiming to reduce hospitalization and mor-tality in CKD patients. By synthesizing recent findings on risk stratification, diagnostic modalities, and individualized treatment—particularly for patients undergoing renal replacement therapy—clinicians can enhance volume management and optimize patient outcomes. Considering the increasing prevalence of chronic kidney disease and associated cardiovascular comorbidities, this review addresses pathogenic mechanisms, diagnostic approaches, pharmacological treatments, and dialytic therapy modifications.

Biomedicines doi: 10.3390/biomedicines14040840

Authors: Sungsu Oh Jeogin Jang Yunseong Ko Hyunsu Lee Seungjin Lim

Background: Liver transplant recipients are highly susceptible to infectious complications due to surgical invasiveness and immunosuppressive therapy, and post-transplant bloodstream infection is associated with substantial morbidity and mortality. Although several prediction models for bloodstream infection have been proposed, most focus on emergency department or general ward populations and rely on black-box approaches. This limits their applicability and clinical interpretability in liver transplant settings. Therefore, this study aimed to develop predictive models for post-transplant bloodstream infection using preoperative and perioperative clinical data and to derive an interpretable risk equation through symbolic regression. Methods: We conducted a retrospective observational study including 245 adult liver transplant recipients treated at a single tertiary center. Clinical and laboratory variables were extracted from electronic medical records and analyzed using standard statistical methods. For prediction tasks, multiple conventional machine learning models were developed and compared with a symbolic regression-based model. Predictive performance and model interpretability were evaluated using discrimination metrics and Shapley Additive Explanations. Results: Post-transplant bloodstream infection occurred in 82 patients (33.4%). In the test set, conventional machine learning models showed modest discriminative performance (area under the curve, 0.53–0.64). The symbolic regression model achieved comparable discrimination (area under the curve, 0.63) while providing transparent, threshold-based risk equations. While conventional models primarily relied on laboratory variables, symbolic regression additionally identified perioperative clinical factors and viral serologic markers as important predictors. Discussion: Although overall predictive performance was modest, symbolic regression highlighted viral serologic markers as potential indicators of immunologic vulnerability, extending beyond standard laboratory predictors. Conclusions: This interpretability-focused approach may inform future risk stratification models incorporating richer perioperative data.

Biomedicines doi: 10.3390/biomedicines14040838

Authors: Felicia Gabriela Beresescu Razvan Marius Ion Adriana-Stela Crisan Andrea Bors

Background: Severe obesity is associated with chronic low-grade inflammation, dysglycemia, and higher periodontitis risk. Sleeve gastrectomy (SG) is now a dominant bariatric procedure and reliably improves weight and metabolic status yet reported oral and periodontal trajectories after surgery remain heterogeneous. Objective: To synthesize SG-centered evidence on periodontal outcomes, oral and gut microbiome remodeling, and mechanistic pathways that may link postoperative physiology to the gut–oral axis. Methods: We conducted a structured narrative review guided by SANRA principles using targeted searches of PubMed/MEDLINE, Web of Science, Scopus, and Embase, complemented by citation chaining of key reviews and mechanistic anchor papers; evidence was organized into clinical, oral microbiome, gut microbiome, and mechanistic gut–oral axis streams and interpreted with a pragmatic evidence hierarchy. Results: Small prospective SG cohorts suggest bleeding on probing (BOP), gingival indices, and sometimes probing depth (PD) may improve in some patients, particularly alongside weight loss, improved glycemic control, and lower systemic inflammatory burden, whereas clinical attachment level (CAL) and longer-term structural trajectories remain mixed; mixed-procedure syntheses also report early deterioration in some settings. Oral microbiome findings after bariatric surgery are site- and time-dependent, and salivary signals do not necessarily mirror subgingival plaque, whereas gut microbiome remodeling and bile acid signaling changes are more consistently reported and provide plausible but indirect mediator candidates. At the same time, reflux, vomiting, salivary changes, diet patterning, medications, and periodontal care can modify or counteract potential periodontal benefits and may increase competing risks such as caries or erosive tooth wear. Conclusions: The SG–gut–oral axis-periodontal pathway is a biologically plausible working hypothesis rather than a proven causal pathway in humans. The present evidence for any periodontal benefit relies mainly on small observational cohorts and is most credibly demonstrated for inflammatory, not structural, endpoints.

Biomedicines doi: 10.3390/biomedicines14040839

Authors: Sofía Tejada Andrés Giglio Maria Aranda Antonia Socias Alberto del Castillo Joana Mena Sara Franco Maria Ortega Yasmina Nieto Marcio Borges-Sa

Background/Objective: Coagulopathy is a hallmark of sepsis, associated with poor outcomes. Although platelet count has commonly been used for risk stratification, its prognostic value has remained limited. This study compared the ability of the International Normalized Ratio (INR) and platelet count to predict in-hospital mortality in septic patients; Methods: A retrospective study was conducted including adult patients diagnosed with sepsis and admitted to Hospital Universitario Son Llàtzer (Spain) between 2006 and 2022. The INR and platelet count at diagnosis were categorized using clinical thresholds. The primary outcome was in-hospital mortality. Results: Among 6433 patients (60.6% females), mortality was 8.8%. Mortality increased from 6.3% (INR ≤ 1.2) to 20.2% (INR 2.0–3.0), slightly decreasing at INR > 3.0 (10.8%). The platelet count showed a weaker association, with the highest mortality observed at <50 × 109/L (24.6%). The combined markers identified a high-risk subgroup with 50% mortality (INR > 3.0 and platelet count < 50 × 109/L). In the full cohort, multivariable analysis confirmed the INR as an independent predictor of mortality (OR 2.183, p = 0.0002), whereas the platelet count was not significant. The model including the INR achieved an AUC 0.746, while adding the platelet count did not improve performance. Conclusions: the INR at diagnosis was a strong and independent predictor of in-hospital mortality, outperforming the platelet count. These findings could support the consideration of the INR in early risk stratification frameworks and highlight the need for prospective validation before integration into sepsis guidelines.

Biomedicines doi: 10.3390/biomedicines14040837

Authors: Lara Batičić Božena Ćurko-Cofek Gordana Taleska Štupica Matej Jenko Marko Zdravković Lea Cofek Antea Krsek Tanja Batinac Danijel Knežević Marino Damić Mia Šestan Aleksandra Ljubačev Maja Šoštarič Vlatka Sotošek

Open-heart surgery with cardiopulmonary bypass (CPB) is a high-risk procedure with significant morbidity and mortality. CPB, tissue injury, blood loss, endotoxemia and ischemia–reperfusion injury induce a pronounced systemic inflammatory response, leading to endothelial glycocalyx (EG) damage and vascular endothelial dysfunction. Consequently, immune cells, reactive oxygen species, and enzymes gain free access to vascular endothelial cells, resulting in their dysfunction and enhancing inflammation, vascular permeability, and microvascular impairment. EG degradation is most commonly assessed by measuring the circulating levels of its degradation products. Additionally, CPB triggers an early inflammatory response that is characterized by the secretion of interleukin (IL)-1β, IL-6, IL-8, tumor necrosis factor alpha, and IL-18, which play roles in initiating the process of EG injury. EG damage is further propagated by the sustained release of cytokines, inhibiting the regeneration of the glycocalyx layer. Heparanase and matrix metalloproteinases are enzymatic pathways involved in cytokine-mediated EG degradation after cardiac surgery, and the balance between the pro- and anti-inflammatory cytokines determines the magnitude and duration of the inflammatory response and EG impairment, which correlates with adverse clinical outcomes, including myocardial dysfunction, acute lung and kidney injury, neurological complications, and prolonged need for intensive care. Thus, identifying patients with an exaggerated cytokine response could potentially provide more personalized therapy based on the circulating biomarkers of EG shedding, and cytokine-directed preservation of EG represents a promising therapeutic strategy in vascular dysfunction prevention during and after open-heart surgery. In this review, we summarize the current knowledge on cytokine-mediated EG impairment following open-heart surgery with CPB.

Biomedicines doi: 10.3390/biomedicines14040836

Authors: Vinay Devulapalli Akash Sathiyamurthi Surabhi Gautam Pallavi Bhattaram

Alpha-Ketoglutarate (AKG), a central intermediate of the tricarboxylic acid cycle, is a crucial metabolic and signaling molecule that connects mitochondrial function with cellular homeostasis, immunological modulation, epigenetic remodeling, and lifespan. While mitochondrial AKG maintains energy metabolism, the nuclear AKG pool influences chromatin remodeling through DNA and histone modifications, which together control hypoxia responses and shape gene expression patterns. This dual role demonstrates AKG’s significance in mediating metabolic state, gene expression, and long-term cellular adaptability. AKG modulates immunological responses, reduces reactive oxygen species (ROS), promotes the polarization of anti-inflammatory macrophages, and suppresses nuclear factor kappa B (NF-κB) activation, thereby reducing chronic inflammatory processes. AKG restricts pro-inflammatory cytokine production, increases extracellular matrix synthesis, and reduces cartilage degradation in arthritic models, suggesting potential therapeutic benefits in autoimmune diseases and joint degeneration. Additionally, AKG affects lifespan in several model organisms, where supplementation enhances metabolic resilience, lowers age-related inflammation, modifies mTOR signaling, and preserves youthful epigenetic profiles. Additionally, because endogenous AKG levels decrease with age, oral supplementation of AKG, especially with calcium and arginine, has drawn attention to its potential benefits in longevity and metabolic health. Thus, AKG is versatile and has encouraging therapeutic promise for cancer, aging, and inflammatory illnesses. However, a lack of human clinical evidence prompts further research to determine ideal dosage, tissue selectivity, and long-term safety. The goal of this review is to critically examine the current mechanistic knowledge related to AKG biosynthesis and breakdown and its future implications in maintaining cellular homeostasis and controlling chronic inflammation.

Biomedicines doi: 10.3390/biomedicines14040835

Authors: Senyang Guo Jianhua Liu Hongmei Zheng Xinhong Wu

Molecular subtype–guided therapy for breast cancer (BC) remains limited in a subset of patients by suboptimal efficacy, acquired resistance, and the presence of “undruggable” targets. Proteolysis-targeting chimeras (PROTACs) represent a targeted protein degradation (TPD) strategy that differs fundamentally from conventional occupancy-driven inhibition. By inducing ubiquitination of a protein of interest and subsequent proteasomal degradation, PROTACs can directly reduce pathogenic protein abundance and potentially abrogate non-catalytic or scaffolding functions, thereby enabling more durable pathway suppression in selected resistance contexts. This review comprehensively summarizes the mechanisms of action, key molecular design elements, and the developmental landscape of PROTACs, and maps target selection and research progress across BC molecular subtypes. In hormone receptor–positive/HER2-negative BC, clinical translation is most advanced for estrogen receptor alpha-directed PROTACs; Phase III evidence indicates biomarker-dependent efficacy, with clearer benefit signals in resistant subgroups such as estrogen receptor 1 mutations, suggesting that the net clinical benefit of TPD is more likely to be realized through precision stratification. In contrast, in solid-tumor settings, including human epidermal growth factor receptor 2 (HER2)-positive BC and triple-negative breast cancer, PROTAC translation is more frequently constrained by an “exposure–selectivity–therapeutic window” trade-off driven by physicochemical liabilities, insufficient tumor penetration, and broad target expression. Accordingly, engineering strategies—such as antibody/aptamer-mediated targeted delivery, stimulus-responsive prodrugs, nanocarriers, and local administration—are emerging as decisive approaches to enable safe and effective clinical implementation. Looking forward, further progress of PROTACs in BC will depend on expanding the spectrum of E3 ubiquitin ligases and recruitment modalities, establishing predictable and dynamically monitorable biomarker systems, optimizing rational combination/sequencing regimens with exposure- and schedule-guided dosing, and advancing scalable manufacturing and quality control capabilities, thereby translating mechanistic advantages of TPD into verifiable precision-therapy applications.

Biomedicines doi: 10.3390/biomedicines14040834

Authors: Wan-Ru Yu Jia-Fong Jhang Yuan-Hong Jiang Hann-Chorng Kuo

Purpose: This study compared the therapeutic efficacy of different bladder monotherapies and multimodal therapy in patients with interstitial cystitis/bladder pain syndrome (IC/BPS). Materials and methods: In total, 190 patients with a confirmed diagnosis of IC/BPS were treated with different bladder therapies. The bladder monotherapies included intravesical platelet-rich plasma (PRP) injection (n = 60), intravesical botulinum toxin A (BoNT-A) injection (n = 33), intravesical hyaluronic acid (HA) instillation (n = 36), and low-energy shock wave (LESW) bladder therapy (n = 61). Multimodal therapy (MMT) was provided to patients who had unsuccessful initial bladder treatment targeting chronic inflammation, urothelial dysfunction, bladder pain, pelvic floor muscle pain, psychological stress, and lower urinary tract dysfunction. The treatment outcome was assessed using self-reported Global Response Assessment scores at 3 months and during the follow-up time points after bladder treatment. Results: Thirty-one patients received MMT. The 3-month success rates of bladder therapy were 55.0% for PRP injection, 57.6% for BoNT-A injection, 50.0% for HA instillation, 46.7% for LESW bladder therapy, and 58.1% for MMT. The success rates of bladder monotherapy decreased after 6 months. However, the success rate of MMT increased at 9 (67.7%) and 12 (73.1%) months. Patients treated with MMT exhibited improvement in glomerulation grade after cystoscopic hydrodistention. Only patients with successful treatment outcomes after MMT had improvement in bladder pain severity and pelvic floor muscle pain parameters. Conclusions: Bladder monotherapy such as PRP injection, BoNT-A injection, HA instillation, and LESW bladder therapy had successful treatment outcomes in patients with IC/BPS. In patients who had unsuccessful initial bladder therapy, the 3-month success rate of MMT was 58.1% and sustained improvement with time, particularly in the improvement of bladder pain and PFM pain severity.

Biomedicines doi: 10.3390/biomedicines14040833

Authors: Apostolos Sioutas Hans Lennart Persson

Background: Chronic obstructive pulmonary disease (COPD) is associated with systemic inflammation and frequent exacerbations, leading to disease progression and increased morbidity. Vitamin D deficiency has been suggested to contribute to COPD inflammation and exacerbations. Aim: This study investigated the association between serum 25-hydroxyvitamin D (25(OH)D) levels, systemic inflammation, and exacerbation frequency among patients with COPD GOLD group E. Methods: A cross-sectional study was conducted on 111 patients with stable COPD. Patients were divided into two groups based on their serum 25(OH)D levels (<50 nmol/L vs. ≥50 nmol/L). Data on exacerbation frequency for the past year, inflammatory markers, spirometric lung function parameters, and symptom burden were collected. Results: Patients with low serum 25(OH)D (<50 nmol/L) had a significantly higher CAT score and level of serum high-sensitivity (hs)-CRP and exhibited significantly more exacerbations compared to those with higher 25(OH)D levels (p < 0.001, p < 0.001, and p < 0.0001, respectively). Furthermore, lower vitamin D levels were associated with higher CAT scores (Pearson’s correlation coefficient, r = −0.30, p < 0.01) and higher serum hs-CRP levels (Spearman’s rank correlation coefficient, r = −0.25, p < 0.01), as well as a higher number of exacerbations (Pearson’s correlation coefficient, r = −0.74, p < 0.0001). Conclusions: Low vitamin D levels are significantly associated with greater symptom burden, elevated hs-CRP, and increased exacerbation frequency, indicating a strong relationship between vitamin D deficiency, systemic inflammation, and disease burden in patients with COPD belonging to GOLD group E. However, due to the cross-sectional design, no causal relationship can be inferred and prospective interventional studies are required to determine whether treating vitamin D deficiency improves clinical outcomes.

Biomedicines doi: 10.3390/biomedicines14040832

Authors: Ravi Dhital Mukut Minz Ranjana Walker Minz Shashi Anand Ritambhra Nada Sarbpreet Singh Deepesh B. Kenwar Ashish Sharma

Background: Renal allograft rejection remains a major challenge in transplantation. Current diagnostic approaches, including biopsies, are invasive and may fail to detect subclinical immune activation, potentially contributing to progressive graft dysfunction. Reliable, non-invasive biomarkers capable of monitoring immune activation and distinguishing rejection phenotypes are therefore needed. Methods: In this retrospective study, we evaluated serum neopterin as a biomarker of immune activation and graft status over 12 months following transplantation. Associations between neopterin levels and immune parameters, including natural killer (NK)-to-CD3+CD16/CD56+ T cell ratios, cytokines (IFN-γ and IL-10), and CD4+CD25+FoxP3+ T cell frequencies, were assessed. A total of 211 first renal allograft recipients were followed longitudinally, including patients with acute rejection (AR) and matched stable graft (SG) recipients. Serum neopterin was quantified by enzyme immunoassay, and immunophenotyping, mRNA expression, and cytokine profiling were performed on peripheral blood samples. Results: Serum neopterin levels were significantly elevated in AR compared to SG recipients, with a threshold of 57 nmol/L distinguishing AR with 81% sensitivity and 80% specificity. While IFN-γ demonstrated higher diagnostic performance in cross-sectional analysis, neopterin showed a more sustained elevation over time, remaining increased in AR recipients even at later post-transplant time points. Neopterin correlated positively with IFN-γ, but not IL-10, and inversely with CD4+CD25+FoxP3+ T cell frequency. NK cells were enriched during rejection, whereas CD3+CD16/CD56+ T cells were more prominent in graft stability. The NK-to-CD3+CD16/CD56+ T cell ratio was highest during acute cellular rejection. Conclusions: Neopterin reflects Th1-associated immune activation in renal allograft recipients and provides a temporally stable, non-invasive marker of immune status. Although it does not outperform IFN-γ levels at the time of rejection, its stability and sustained elevation suggest a complementary role in longitudinal monitoring. Integration of neopterin with immune parameters, including cytokine profiles and cellular subsets, may enhance the assessment of graft immunological status and support clinical decision-making.

Biomedicines doi: 10.3390/biomedicines14040831

Authors: Almira Akparova Gaukhar Kurmanova Gulzhakhan Omarova Almagul Kurmanova Moldir Zhunisbek Magripa Bapaeva Zhamilya Zhankina Sholpan Sadykova Amina Abdrakhmanova Adema Samadin

Mucins are highly glycosylated proteins that form the structural basis of mucus and represent a key component of innate immunity at mucosal surfaces, particularly in the respiratory tract. Beyond their mechanical barrier function, mucins actively participate in pathogen trapping, regulation of mucociliary clearance, modulation of inflammatory responses, and maintenance of epithelial homeostasis. Dysregulation of mucin synthesis, composition, or transport contributes to mucus hypersecretion, impaired airway clearance, and chronic inflammation in respiratory diseases such as asthma, chronic obstructive pulmonary disease, and cystic fibrosis. This review summarizes current insights into mucin biology, including their biosynthesis, structure, classification, and regulation, with emphasis on the gel-forming mucins MUC5AC and MUC5B. The role of mucins in mechanical protection, host–pathogen interactions, control of inflammation, and coordination of innate immune responses is reviewed. Attention is given to the interplay between mucins, immune cells, and microbial communities in maintaining airway barrier integrity. The article further examines mucoactive therapeutic strategies aimed at restoring mucus barrier function. Expectorants, mucolytics, mucoregulators, and mucokinetic agents are reviewed with respect to their mechanisms of action and clinical relevance. Established drugs, including N-acetylcysteine, carbocysteine, dornase alfa, ambroxol, and hypertonic solutions, are considered alongside emerging molecular targets such as NF-κB-dependent regulation of mucin expression, calcium-activated chloride channels, MARCKS-mediated mucin exocytosis, purinergic signaling pathways, and NO/cGMP signaling. Non-pharmacological approaches, including airway clearance techniques and respiratory rehabilitation, are covered concisely. Conclusions: Overall, this review highlights mucins as dynamic regulators of innate immunity and underscores the need for mechanism-based, personalized mucoactive therapies to improve outcomes in chronic inflammatory airway diseases.

Biomedicines doi: 10.3390/biomedicines14040830

Authors: Sean Hsiang-Ting Chen Tsung-You Tsai Rodney Cheng-En Hsieh Kai-Ping Chang Chung-Jan Kang Yi-An Lu Pei-Wei Huang Miao-Fen Chen Chien-Yu Lin Shanli Ding Ngan-Ming Tsang Wen-Hsin Lu Wing-Keen Yap Alex Chia-Hsin Lin

Background/Objectives: We aim to investigate whether tracking pan-immune–inflammation value (PIV) dynamics during radiotherapy (RT) can inform real-time prognosis in patients with head and neck cancer (HNC). Methods: We retrospectively reviewed the medical records of patients with HNC who received RT at our institution between 2005 and 2013. Temporal changes in the PIV throughout the RT were evaluated using the Friedman test and Wilcoxon signed-rank test. The PIV dynamics were quantified using PIV ratios, defined as the PIV at three distinct time points (PIV-2, PIV-4, and PIV-6) during treatment divided by the pretreatment PIV (PIV-0). Overall survival (OS) and progression-free survival (PFS) served as the primary and secondary endpoints analyzed. Results: A total of 676 patients with HNC were enrolled, with a median follow-up of 8.1 years. The PIV demonstrated a continuously ascending trend over time, with the most dramatic increase occurring six weeks after the start of RT. Compared with patients with a low PIV ratio at six weeks (PIV-6/PIV-0), those with a high PIV ratio showed more favorable survival outcomes (five-year OS: 58.9% versus 70.8%, p = 0.002; five-year PFS: 62.0% versus 71.1%, p = 0.013). The subgroup analyses yielded consistent results. Notably, the real-time risks of death and recurrence changed in parallel with the PIV dynamics. Multivariate analysis confirmed PIV-6/PIV-0 as an independent prognostic factor for both OS and PFS. Conclusions: Monitoring longitudinal PIV dynamics may assist in forecasting the OS and PFS in patients with HNC being treated with RT, thus enabling individualized, risk-adapted treatment management.

Biomedicines doi: 10.3390/biomedicines14040826

Authors: Mi Eun Kim Jun Sik Lee

Interleukin-10 (IL-10) is a key immunoregulatory cytokine that suppresses inflammatory gene transcription in myeloid cells through signal transducer and activator of transcription 3 (STAT3). In Alzheimer’s disease and neuroinflammation, microglia express IL10ra and exhibit STAT3 Tyr705 phosphorylation following IL-10 stimulation, indicating IL-10 receptor-dependent STAT3 activation. Recent studies demonstrate that IL-10 induces promoter-selective STAT3-dependent transcriptional regulation in microglia through chromatin-associated mechanisms, whereas gp130-dependent cytokines activate STAT3 to induce transcription of defined target genes, including Socs3 and Ccl5. Following IL-10 receptor activation, STAT3 binds regulatory regions of inflammatory genes, including Il1b, Tnf, Il6, and Nlrp3, with reduced RNA polymerase II and NF-κB binding. IL-10-dependent transcriptional repression involves formation of a nuclear SHIP1–STAT3 complex, localization of histone deacetylase (HDAC)1 and HDAC2 to H3K4me1-enriched enhancer regions, reduced H3K27ac, and decreased chromatin accessibility at regulatory regions of inflammatory genes. IL-10-activated STAT3 induces Socs3, which regulates JAK1 and TYK2 activity and STAT3 phosphorylation. Impairment of IL-10 receptor signaling in microglia is associated with increased inflammatory gene expression, enhanced inflammasome-related transcription, demyelination, and amyloid accumulation. This review focuses on IL-10–STAT3-dependent transcriptional regulation in microglia, including receptor signaling, chromatin-associated mechanisms, and disease-associated gene expression in Alzheimer’s disease and neuroinflammation.

Biomedicines doi: 10.3390/biomedicines14040829

Authors: Wei Xu Yang Yang Fuqiang Li Can Li Gaojun Tang Baofang Zhang Mingliang Cheng

Objectives: This study aims to investigate the potential molecular mechanisms by which α-hederin modulates HSC activation to alleviate liver fibrosis. Methods: An in vitro model of liver fibrosis was established by inducing LX-2 cells with TGF-β1. These cells were then treated with α-hederin (10 μg/mL) before undergoing phenotypic analysis and molecular-level detection. A mouse model of liver fibrosis induced by CCl4 was established in vivo to further evaluate the expression levels of fibrosis markers, including TRIM38. Results: In TGF-β1-induced liver fibrosis in LX-2 cells, α-hederin treatment significantly inhibited HSCs activation, as evidenced by down-regulation of α-SMA and suppressed proliferation capacity. At the same time, α-hederin significantly reduced the levels of COL1A1, COL3A1, fibronectin, and MMP-2. Transcriptome sequencing analysis revealed that α-hederin treatment significantly upregulated TRIM38 expression. Differentially expressed genes (DEGs) were significantly enriched in endoplasmic reticulum stress-related pathways. TRIM38 up-regulation inhibits HSC activation and proliferation, reducing the expression of ERS marker proteins (GRP78, p-PERK, and CHOP); Co-IP experiments further confirmed that TRIM38 and GRP78 interact directly. Further rescue experiments demonstrated that TRIM38 knockdown significantly attenuated the inhibitory effects of α-hederin on these processes. In a CCl4-induced mouse model of liver fibrosis, α-hederin (4 mg/kg) significantly reduced the liver index and serum ALT and AST levels, improved histopathological damage to the liver, upregulated TRIM38 expression in liver tissue, and inhibited the endoplasmic reticulum stress response (ERS). Conclusions: α-hederin exerts its anti-fibrotic effect by upregulating TRIM38, thereby alleviating endoplasmic reticulum stress and ultimately inhibiting the activation and proliferation of HSCs.

Biomedicines doi: 10.3390/biomedicines14040828

Authors: Junhong Li Guihua Yu Shiheng Wang Zekai Zhang Yu Wen Luting Yu Xin Gan Hao Kang Jinming Zhang Lu He

Background: Osteoarthritis (OA) is a multifactorial disease, including inflammation, autophagy and senescence. Published work has indicated that empagliflozin (EMP) exhibits robust anti-inflammatory and anti-senescence effects, while its role in autophagy appears paradoxical. Here, we aim to identify the chondroprotective effect of EMP on OA. Methods: An OA model was established both in vitro, by stimulating primary chondrocytes (isolated from C57BL/6J mice) with IL-1β, and in vivo, by performing (Destabilized medial meniscus) DMM surgery on C57BL/6J mice. (Western blot) WB and (quantitative real-time polymerase chain reaction) qRT-PCR analysis were employed to detect the gene expression. (Immunofluorescence) IF staining was employed to detect the expression and location of target protein. SA-β-gal staining was employed to evaluate cellular senescence. Autophagic flux was assessed using a GFP-RFP-LC3 adenoviral vector. Network pharmacology was applied to identify potential pathways for experimental validation. The effects of EMP in vivo were evaluated by μ-CT, histological and (Immunohistochemistry) IHC staining. Results: EMP promoted anabolism, inhibited the inflammatory response and catabolism in IL-1β stimulated chondrocytes. EMP enhanced autophagic activity and attenuated senescent phenotype in vitro. Mechanistically, EMP regulated the PI3K/Akt/mTOR and AMPK pathways. The chondroprotective effects of EMP were reversed by (3-methyladenine) 3-MA. EMP also ameliorated OA-related phenotype in DMM models. Compared with (Kartogenin) KGN, EMP showed more pronounced suppression of inflammatory and catabolic markers, while both compounds similarly promoted anabolic marker expression. Conclusions: These in vitro and in vivo data collectively indicates that EMP can alleviate OA both in IL-1β stimulated chondrocytes and DMM induced models. Beyond its established role in diabetes management, EMP is evaluated in the context of OA, emerging as a novel and promising therapeutic agent for OA.

Biomedicines doi: 10.3390/biomedicines14040827

Authors: Hongkai Dai Cheng Li Bin Yang Qianrui Huang Xiao Ran Yingfang Zheng Yongsheng Li

Background: Sepsis-induced acute lung injury (ALI) is a life-threatening condition with limited therapeutic options. Neutrophil extracellular traps (NETs) contribute to its pathogenesis. This study investigated whether polydatin (PD) protects against septic ALI by inhibiting NETs via the Nrf2/HO-1 pathway. Methods: A cecal ligation and puncture (CLP)-induced septic ALI mouse model and an LPS-stimulated neutrophil model were established. Lung injury was assessed by histology, lung wet/dry ratio, BALF protein, and inflammatory cytokines. Endothelial junction proteins and NETs markers were examined by Western blot, immunofluorescence, and SYTOX Green staining. Nrf2/HO-1 pathway activation and ML385 inhibitor experiments were performed for mechanistic validation. Results: PD dose-dependently attenuated lung injury, preserved endothelial junction proteins (ZO-1, VE–cadherin, occludin), and suppressed NETs formation in vivo. In vitro, PD activated Nrf2/HO-1, promoted Nrf2 nuclear translocation, reduced ROS, and inhibited LPS-induced NETs. These effects were abrogated by ML385, confirming pathway specificity. Conclusions: PD mitigates septic ALI by inhibiting NETs formation. In vitro mechanistic studies further suggest that this effect is mediated through activation of the Nrf2/HO-1 antioxidant pathway, positioning PD as a potential therapeutic candidate for sepsis-induced ALI.

Biomedicines doi: 10.3390/biomedicines14040825

Authors: Andrei Mihordea Mihai Puiu Gelu Simu Ioan-Alexandru Minciuna Radu Rosu Gabriel Gusetu Dana Pop Gabriel Cismaru

Catheter ablation has become a cornerstone therapy for atrial fibrillation, with pulmonary vein isolation as its mechanistic foundation. Radiofrequency ablation and cryoablation, the two established thermal technologies, have demonstrated robust efficacy across multiple randomized trials but remain limited by collateral tissue injury inherent to heat- or cold-mediated lesion formation. Pulsed field ablation has recently emerged as a novel non-thermal energy source based on irreversible electroporation, offering myocardial-selective injury with relative sparing of adjacent structures. This review synthesizes evidence across three complementary domains: fundamental studies; preclinical evidence; and clinical data supporting radiofrequency ablation, cryoablation, and pulsed field ablation for atrial fibrillation. We summarize mechanistic differences in lesion formation, key animal studies that established safety and efficacy profiles, and pivotal randomized clinical trials, including recent head-to-head comparisons and meta-analyses of randomized controlled trials. By synthesizing these levels of evidence, the review aims to place recent clinical results into a mechanistic and translational context. Available evidence demonstrates that pulsed field ablation achieves rhythm-control efficacy comparable to radiofrequency and cryoablation while offering procedural efficiency and a potentially improved safety profile. However, long-term durability data and broader experience remain limited. Understanding the strengths and limitations of each ablation modality is essential for informed clinical decision-making as non-thermal ablation technologies enter routine practice.

Biomedicines doi: 10.3390/biomedicines14040824

Authors: Ching-Chang Cho Gow-Chin Yen Hsin-Yi Lee Wei-Tang Chang Li-You Chen Chin-Lin Hsu

Background/Objectives: Obesity represents a critical risk factor for various chronic illnesses and metabolic dysfunctions, underscoring the urgency of identifying safe, food-based interventions to curb fat over-accumulation. Mesona procumbens Hemsl. (Hsian-tsao) is a traditional Chinese herb known for its antioxidant and health-promoting properties; however, it remains unclear how its phenolic acid profiles contribute to anti-obesity activity. This research explored the anti-adipogenic potential of various Hsian-tsao ethanol extracts, focusing on how their phenolic profiles influence lipid suppression in 3T3-L1 adipocytes. Methods: Ethanol extracts prepared using different ethanol concentrations were analyzed for total polyphenol content, antioxidant capacity, and phenolic acid profiles. Adipocytes were exposed to 0, 100, and 250 μg/mL of Hsian-tsao ethanol extract for 48 h duration to monitor changes in cell count and intracellular triglyceride levels. Results: Among all fractions, the 40% ethanol extract (40EEHT) possessed the most robust antioxidant capacity and highest polyphenol content, specifically showing enriched levels of caffeic acid, p-coumaric acid, and total phenolic acids. Notably, while 40EEHT influenced cell density at certain concentrations, it significantly and specifically reduced intracellular triglyceride content, indicating a potent inhibitory effect on lipid storage independent of changes in cell number. Comparative analysis using phenolic acid standards revealed that caffeic acid exerted the strongest inhibitory effect on lipid accumulation, suggesting that it is a key contributor to the anti-adipogenic activity of 40EEHT. Conclusions: Collectively, these findings demonstrate that phenolic acid profiles, particularly caffeic acid enrichment, critically contribute to the potential anti-adipogenic effects of specific ethanol extracts of M. procumbens. Therefore, Hsian-tsao ethanol extracts represent a promising natural source for the development of functional ingredients targeting obesity and related metabolic disorders.

Biomedicines doi: 10.3390/biomedicines14040823

Authors: Leonie Ripp Dennis Kätzel

Background: Translational circuit neuroscience delivers many candidate neurons whose manipulation could ameliorate psychiatric symptoms. However, the translation of these cellular targets into molecular targets—proteins selectively expressed in those neurons that could be pharmacologically manipulated for treatment—remains scarce. To what extent such a translation is possible or is actually impeded by a lack of highly cell type-specific expression of druggable proteins is unknown. Methods: We performed combinatorial differential expression analysis for over 7200 putatively druggable genes (Illuminating the Druggable Genome database) on large-scale single-cell RNAseq datasets from mouse and human cortex (Allen Institute Cell Types Database) to identify selectively expressed genes in important cellular candidates: several pyramidal cell types and parvalbumin, somatostatin and VIP interneurons of the prefrontal and anterior cingulate cortex and hippocampus in mice, and the cingulate cortex in humans. Results: We identified dozens of targets, including some with psychiatric relevance and/or suitability to modulate neural activity, like ion channels, GPCRs and transporters. However, none of them were expressed with absolute specificity in any of the analysed target cell types but only stood out in some comparisons, not others. Generally, results depended strongly on selectivity criteria: less conservative approaches (such as moderate p-value adjustment or grouping of contrast cell sets) yielded more targets, whereas the introduction of additional plausible constraints (difference in proportion of expressing cells, beta; absence of expression in contrast cell type) drove numbers towards zero. Generally, interneurons showed more selectively expressed targets in comparison to cells of the same region compared to excitatory ones (intra-regional comparisons), whereas the reverse was found in inter-regional contrasts comparing the same cell type across regions. Conclusions: The lack of high selectivity in the expression of genes encoding druggable targets constitutes a principal biological limit for manipulating cortical neurons of one type, specifically to leverage therapeutic action. While, currently, this conclusion is limited to the investigated neocortical and hippocampal regions, it highlights the need to develop biological heuristics for identifying targets expressed with relative specificity.

Biomedicines doi: 10.3390/biomedicines14040821

Authors: Thanh Van Le Vinh Thanh Tran Ai Thi Kim Le Linh Ha Khanh Duong

Background/Objectives: Iron deficiency (ID) is a critical comorbidity in chronic heart failure (CHF) that impairs myocardial energy metabolism and clinical outcomes. Despite its significance, data regarding ID prevalence in Southeast Asian CHF populations remain insufficient. This study aimed to determine the prevalence of ID and identify its independent clinical correlates among CHF patients at a leading tertiary hospital in Vietnam using high-precision automated diagnostic platforms. Methods: A descriptive cross-sectional study was conducted on 138 adult CHF patients. Serum iron and ferritin were quantified using photometric and electrochemiluminescence immunoassay (ECLIA) methods on cobas c702 and e602 systems. ID was defined according to 2021 ESC guidelines (ferritin < 100 ng/mL or ferritin 100–299 ng/mL with TSAT < 20%). Multivariable logistic regression identified independent factors associated with ID. Results: The overall ID prevalence was 75.4%, consisting of functional (39.9%) and absolute ID (35.5%). A significant gender disparity was observed (p = 0.0326): absolute ID was more prevalent in females (44.4%), while functional ID predominated in males (47.4%). Multivariable analysis revealed that NT-proBNP (aOR = 2.29; 95% CI: 1.13–4.66; p = 0.021) and C-reactive protein (aOR = 2.17; 95% CI: 1.07–4.43; p = 0.031) were independent correlates of ID status. The association with anemia was borderline non-significant (p = 0.055). Conclusions: ID is nearly ubiquitous among Vietnamese CHF patients in this tertiary setting, driven by hemodynamic severity and systemic inflammation. These findings advocate for integrating routine iron status screening into the standard diagnostic workup for all CHF patients, regardless of hemoglobin levels.

Biomedicines doi: 10.3390/biomedicines14040819

Authors: Tanya Sapundzhieva Lyubomir Sapundzhiev Plamen Todorov Martin Mitev Anastas Batalov

Aims. To investigate the relationship between ultrasound (US)-detected parenchymal abnormalities in the major salivary glands (MSG), joint and tendon inflammation, and systemic disease activity in patients with primary Sjögren’s syndrome (pSS). Patients and methods. This cross-sectional, multicenter study enrolled 60 patients with pSS and 20 healthy controls (HCs). Systemic disease activity was evaluated using the EULAR Sjögren’s Syndrome Disease Activity Index (ESSDAI), while symptom burden was assessed with the EULAR Sjögren’s Syndrome Patient Reported Index (ESSPRI). MSG evaluation included bilateral gray-scale (GS) and power Doppler (PDUS) assessment of the parotid and submandibular glands using a semi-quantitative 0–3 scoring system. Musculoskeletal ultrasound (MSUS) assessment comprised bilateral examination of the wrists, second to fifth metacarpophalangeal (MCP) and proximal interphalangeal (PIP) joints, the fourth extensor wrist compartment, and the flexor tendons of the second to fifth fingers for GS and PD-detected synovitis and tenosynovitis, also scored semi-quantitatively. Recorded outcomes included GS and PD synovitis scores, total synovitis score, tenosynovitis score, GS and PD glandular scores, and total glandular score. Results. Synovitis was most frequently detected in the wrists, followed by the second PIP joint. Subclinical synovitis—defined as a GSUS synovitis score > 0 in a joint without clinical swelling—was detected in 66.7% (n = 28) of patients with pSS. No significant correlations were found between joint US scores and salivary gland US scores. ESSPRI showed moderate positive correlations with both the GS synovitis score (p = 0.002) and the total synovitis score (p = 0.003), as well as significant positive correlations with all salivary gland US scores: GS (p < 0.001), PD (p = 0.002), and total glandular score (p < 0.001). ESSDAI demonstrated only a weak positive correlation with the GS salivary gland score (p = 0.030). Conclusions. In patients with pSS, the extent of US-detected MSG parenchymal abnormalities does not reflect systemic disease activity and does not correlate with US-detected joint synovitis. In contrast, patient-reported symptom burden is associated with both joint inflammation and MSG parenchymal changes on US. Larger studies are needed to further define the role of salivary gland and joint US in evaluating disease activity in pSS.

Biomedicines doi: 10.3390/biomedicines14040822

Authors: Marco Fiore Gianluigi Cosenza Francesco Maria Romano Vincenzo Pota Pasquale Sansone Francesco Coppolino Lucio Selvaggi Francesco Selvaggi Maria Caterina Pace

Background/Objectives: Abdominal sepsis remains a major contributor to morbidity and mortality among surgical and critically ill patients worldwide. Timely diagnosis is frequently hindered by the overlapping clinical and biochemical features of postoperative inflammatory responses and evolving intra-abdominal infections, which may resemble systemic sepsis. Conventional biomarkers, including C-reactive protein (CRP) and procalcitonin (PCT), are widely implemented in clinical practice but demonstrate suboptimal specificity in differentiating infectious from sterile inflammatory conditions in the early postoperative phase. Presepsin (soluble CD14 subtype, sCD14-ST), a circulating fragment released during monocyte–macrophage activation in response to bacterial endotoxins, has emerged as a biomarker reflecting innate immune engagement. This review aims to critically evaluate the current evidence regarding the diagnostic accuracy, prognostic relevance, and potential clinical role of presepsin in abdominal sepsis. Methods: A comprehensive narrative review of the biomedical literature was performed using MEDLINE (via PubMed) and supplementary academic sources. Studies assessing the diagnostic performance, prognostic associations, and clinical applicability of presepsin in abdominal infections, postoperative infectious complications, and sepsis were systematically examined. Where available, comparative analyses with established biomarkers such as CRP and PCT were evaluated to contextualize its incremental value within existing diagnostic frameworks. Results: The accumulated evidence indicates that presepsin concentrations increase early during bacterial infections and correlate with validated severity indices, organ dysfunction scores, and mortality outcomes. Across multiple surgical and intensive care settings, presepsin demonstrated moderate-to-high diagnostic performance, frequently comparable to and occasionally exceeding that of traditional inflammatory biomarkers, particularly in distinguishing septic from non-septic inflammatory states. Moreover, dynamic changes in circulating levels appear to provide additional prognostic information and may support longitudinal clinical assessment. Nonetheless, substantial heterogeneity in study design, patient populations, sampling strategies, and reported cut-off values limits direct cross-study comparability and constrains definitive clinical recommendations. Conclusions: Presepsin represents a biologically plausible and clinically promising biomarker for the early identification and risk stratification of abdominal sepsis. Although current findings are encouraging, further large-scale, methodologically standardized prospective investigations are required to define optimal diagnostic thresholds and to clarify their role within multimodal biomarker strategies in contemporary sepsis management.

Biomedicines doi: 10.3390/biomedicines14040820

Authors: Wouter T. Moest A. Lianne Messchendorp Helma Dolmans Cynthia Konijn-Janssen Stan van den Eijnden Milou van Bruchem Ineke Tieken Maarten H. L. Christiaans Arjan D. van Zuilen Marcia M. L. Kho Irma Stijnman Frederike J. Bemelman Jan-Stephan S. Sanders Mariet C. W. Feltkamp Aiko P. J. de Vries Joris I. Rotmans

Background: BK polyomavirus (BKPyV) infection is of notable concern in kidney transplant recipients, as it can cause BKPyV-associated nephropathy (BKPyVAN). Currently, there is no effective treatment for BKPyV infection, underscoring the need for preventive strategies. There is emerging evidence that donor-derived BKPyV plays a role in the development of BKPyV DNAemia. To further explore this hypothesis, we conducted a retrospective, multi-center cohort study to evaluate the risk of developing BKPyV DNAemia in kidney recipient pairs sharing the same donor. Methods: At the Leiden University Medical Center (LUMC), deceased donor kidney transplant recipients (2011–2021) were identified and classified according to the occurrence of BKPyV DNAemia within the first year post-transplantation. For each recipient, the contralateral kidney recipient from the same donor was identified through national transplant registries. Cox regression was used to assess whether BKPyV DNAemia in the LUMC recipient was associated with an increased risk of BKPyV DNAemia in the contralateral recipient. Results: Among 117 recipient pairs, BKPyV DNAemia was more frequent when the contralateral recipient was affected (28.8% [15/52]), compared with pairs in which the contralateral recipient remained unaffected (10.8% [7/65], p = 0.013). Multivariable Cox regression analysis confirmed this increased risk (HR 4.9, 95% CI: 1.8–13.6; p = 0.002). Conclusions: This study shows a significantly increased risk of BKPyV DNAemia in recipients of deceased donor kidneys when the contralateral kidney recipient develops BKPyV DNAemia. These findings highlight the influence of donor-derived factors in BKPyV transmission in kidney transplantation.

Biomedicines doi: 10.3390/biomedicines14040818

Authors: Anna Truda Angela Cordella Ilenia De Leo Armando Di Palo Roberta Iorio Simona Marino Roberto La Rocca Claudia Collà Ruvolo Nicoletta Potenza Maria Ravo Giovanna Marchese

Background: Familial prostate cancer (PCa) accounts for nearly 20% of all PCa cases and is associated with increased genetic susceptibility and earlier disease onset. However, early detection and risk stratification in genetically predisposed individuals remain challenging. Circulating cell-free DNA (cfDNA) provides a minimally invasive source of tumor-derived genomic and epigenomic information. Integrating multi-omic cfDNA analyses may enhance the discovery of biomarkers relevant to familial PCa biology. Methods: We conducted a pilot feasibility study employing whole-genome, strand-specific sequencing of cfDNA from eight patients with familial PCa. A unified analytical pipeline was used to jointly profile genomic alterations and epigenomic features. Variant calling, methylation mapping, and allele-specific methylation (ASM) analysis were performed to identify somatic mutations, characterize epigenetic dysregulation, and explore potential interactions between genetic and epigenetic mechanisms. Results: Sequencing revealed 18,878 genetic variants, including 2276 potentially pathogenic alterations. We identified 26 recurrent high-impact mutations, such as stop-gain and start-loss variants, in genes including MUC4, MCM9, and SKA3. Epigenomic profiling demonstrated widespread gene-specific hypermethylation, consistent with transcriptional repression in these loci. ASM events were detected in TTC22, TEX51, WDR89, LAIR2, and SKA3, suggesting coordinated interactions between somatic variation and epigenetic regulation in familial PCa. Conclusions: This proof-of-concept study highlights the feasibility and potential of integrating whole-genome and epigenome profiling of cfDNA to decode the molecular architecture of familial prostate cancer. By jointly capturing genomic alterations and epigenetic signatures, including allele-specific methylation, this multi-omic liquid biopsy approach supports a high-resolution exploration of biologically relevant molecular features. Moreover, this integrated profiling strategy provides a minimally invasive and clinically scalable tool that may substantially improve risk assessment. These findings offer a promising foundation for future validation studies in larger cohorts, with the aim of advancing multi-omic cfDNA analysis as a next-generation technology in the field of precision oncologic epigenetics.

Biomedicines doi: 10.3390/biomedicines14040817

Authors: Fahad F. Almutairi Jaber H. Alsalah Husam K. Alzubaidi Mohammad Mustafa Yasser Bawazir Khalid Bahamdein Hassan Balubaid Haifa Alnahdi Khalid Khashoggi Bander Almutairi Shoaa Shetewi Fahad Alrwaithi Shadi Abushaheen Dana Attraji Sara Alsaylani Huda Alamri Rawan Abdeen Hamzah H. Ahmed Mohammad Khalil

Background: Rheumatoid arthritis (RA) has been linked to increased cardiovascular risk; however, whether RA independently contributes to subclinical atherosclerosis remains unclear. This study aimed to evaluate carotid intima–media thickness (IMT) and carotid narrowing in RA patients compared with controls and to examine their associations with conventional cardiovascular risk factors. Methods: A total of 73 RA patients and 78 healthy controls underwent carotid Doppler ultrasonography to assess IMT and carotid narrowing. Non-parametric analyses were used for between-group comparisons, and associations with clinical variables were evaluated within the RA cohort. Results: The median age was 55 years (IQR: 43–63) in the RA group and 61 years (IQR: 51–68) in controls (p = 0.012). No significant differences were observed in median right CCA IMT (RA: 0.070 mm [IQR: 0.060–0.081] vs. controls: 0.068 mm [IQR: 0.050–0.076]; p = 0.619) or left CCA IMT (RA: 0.065 mm [IQR: 0.051–0.079] vs. controls: 0.065 mm [IQR: 0.050–0.074]; p = 0.701). The prevalence of carotid narrowing was also comparable between groups (right CCA: 15.1% vs. 11.5%, p = 0.633). Within the RA cohort, age was positively correlated with right CCA IMT (Spearman’s rho = 0.375, p = 0.001), and diabetes mellitus was associated with a higher prevalence of right CCA narrowing (34.8% vs. 6.0%, p = 0.003). Conclusions: Carotid IMT and narrowing were not significantly different between RA patients and controls. In this cohort, age and diabetes mellitus were more strongly associated with subclinical carotid atherosclerosis than RA status itself. These findings emphasize the importance of comprehensive cardiovascular risk assessment in RA patients, particularly focusing on traditional risk factors.

Biomedicines doi: 10.3390/biomedicines14040816

Authors: Oluwagbenga Dada Shikshita Singh Francheska Sumadchat Madison Lather Benjamin Brooks JuliAnne E. Allgood

Glioblastoma (GBM) remains the most aggressive primary malignant brain tumor in adults, with poor survival largely driven by diffuse cellular infiltration, profound heterogeneity, and near-universal recurrence following standard therapy. Although maximizing the extent of resection is a key determinant of patient outcome, current clinical imaging modalities lack the spatial resolution necessary to detect microscopic tumor invasion and therapy-resistant cell populations. Emerging in vivo imaging technologies capable of cellular and near-single-cell resolution have therefore become a major focus in preclinical neuro-oncology research, with growing relevance for surgical guidance, treatment adaptation, and translational discovery. This review evaluates multiple optical imaging modalities, including multi-photon microscopy, near-infrared II fluorescence imaging, bioluminescence imaging, photoacoustic imaging, optical coherence tomography, confocal laser endomicroscopy, Raman spectroscopy, autofluorescence microscopy, and fluorescence macroscopy with a focus on their ability to detect residual GBM cells. Despite significant advances, these approaches remain constrained by limitations in molecular target availability, probe delivery across the blood–brain barrier, and signal variability within heterogeneous tumor regions. The biological complexity of GBM further challenges detection, as residual tumor cells are spatially dispersed and phenotypically diverse, limiting the effectiveness of single-marker or single-modality strategies. Together, these findings highlight the need for integrated, biologically informed imaging approaches to improve detection of residual disease and guide surgical decision making.

Biomedicines doi: 10.3390/biomedicines14040815

Authors: Dana-Emanuela Pitic (Coţ) Ramona-Amina Popovici Codruţa-Eliza Ille Ioana-Cristina Talpoş-Niculescu Adelina Chevereşan Daniel Pop Alexandra-Ioana Dănilă Emilia Daliana Muntean Iasmina Denisa Boantă Andreea Kis Ciprian Stroia

Background/Objectives: Oral diseases remain among the most prevalent noncommunicable conditions worldwide, with biofilm-driven dysbiosis playing a central role in dental caries, gingivitis, periodontitis, and oral candidiasis. Curcumin has attracted considerable interest because of its anti-inflammatory, antioxidant, antimicrobial, and regenerative properties. However, its clinical use remains limited by poor water solubility, chemical instability, rapid metabolism, and low bioavailability. This review aimed to provide a comprehensive analysis of curcumin-based nanoformulations for oral health applications, with emphasis on their mechanistic actions, antibiofilm activity, and translational relevance. Methods: This review examined representative nanocarrier systems developed for curcumin delivery in oral health. These included polymeric nanoparticles, nanomicelles and nanoemulsions, solid lipid nanoparticles and nanostructured lipid carriers, nanogels, hydrogels, mucoadhesive films, and metallic or hybrid nanosystems. The analysis focused on molecular mechanisms of action, antimicrobial and antibiofilm effects against major oral pathogens, and key translational challenges. Results/Findings: Across the reviewed studies, nanoformulations consistently improved curcumin solubility, stability, tissue penetration, mucosal retention, and controlled release. Mechanistically, they enhanced anti-inflammatory activity through inhibition of nuclear factor kappa B (NF-κB), strengthened antioxidant defenses via the nuclear factor erythroid 2-related factor 2/heme oxygenase-1 (Nrf2/HO-1) axis, supported tissue repair and osteogenic responses, disrupted oral biofilms, and modulated local immune responses. Antimicrobial activity was reported against Streptococcus mutans, Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, and Candida albicans, with reduced exopolysaccharide production, impaired adhesion, and improved biofilm penetration. Conclusions: Curcumin-based nanoformulations represent promising adjunctive platforms for oral healthcare. However, their clinical translation still requires improved stability in the oral-environment standardized manufacturing and characterization, rigorous safety evaluation, and well-designed controlled clinical studies.

Biomedicines doi: 10.3390/biomedicines14040814

Authors: Da Miao Yao Ge Zhengye Liu Ziqi Wan Haotian Chen Xiaoyin Bai Jiarui Mi

Background: The neutrophil-to-lymphocyte ratio (NLR) is a simple biomarker that reflects the balance between innate immune response and adaptive immunity. Currently, the genetic basis and clinical implications of NLR in relation to inflammatory gastrointestinal diseases have not been extensively explored. Methods: We carried out a genome-wide association study (GWAS) on European individuals from the UK Biobank to detect genetic variants related to NLR, followed by post-GWAS analyses including colocalization analysis, transcriptome-wide association studies (TWAS), and LD score regression. Logistic regression, Cox regression, and gene–environment interaction analysis were used to evaluate the impact of NLR polygenic risk scores (PRS) on inflammatory gastrointestinal disease risks. Results: GWAS of 395,442 Europeans identified 306 genomic regions (731 lead SNPs) associated with NLR, mapping to 1542 genes enriched for immune pathways. Colocalization revealed shared genetic signals with TWAS prioritization of 59, 19, 14, 22 and 28 genes in the whole blood, spleen, terminal ileum, transverse colon and sigmoid colon, respectively. LD-score regression showed significant positive genetic correlations with CD (rg = 0.132), coeliac disease (rg = 0.124), peptic ulcer (rg = 0.138) and duodenal ulcer (rg = 0.220). One-SD increase in NLR PRS predicted higher risk of IBD (OR = 1.05, 95% CI 1.03–1.08), Crohn’s disease (OR = 1.06, 1.02–1.10), ulcerative colitis (OR = 1.05, 1.02–1.08) and coeliac disease (OR = 1.07, 1.03–1.11). Restricted cubic splines demonstrated non-linear relationships of NLR PRS for IBD, CD and UC. Gene environment analyses showed smoking and diabetes amplified the risks, while cardioprotective diet, oily fish intake and polyunsaturated fatty acid level attenuated NLR PRS-associated risk in IBD (mainly CD). Conclusions: Our study delineates the polygenic basis of NLR and establishes its genetic correlation with inflammatory gastrointestinal diseases, offering a genetically informed indicator for disease risk stratification with potential utility in population-level prevention strategies.

Biomedicines doi: 10.3390/biomedicines14040813

Authors: Hsin-Yu Chiang Ssu-Han Huang Tien-Yuan Wu Yen-Chen Tung Yung-Lin Chu Hsiao-Chi Wang Guor-Jien Wei Zheng-Yuan Su

Background/Objectives: Many chronic diseases, including cancer, can be developed in conjunction with excessive intracellular oxidative stress and persistent inflammation. The importance of preventive strategies is highlighted by the potential of phytochemical interventions to mitigate these diseases. The purpose of this study was to investigate how Citrus depressa leaf (CDL) extracts can prevent 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced carcinogenesis in JB6 P+ mouse skin epidermal cells. Methods: CDL extracts were prepared and characterized for their phenolic and flavonoid contents. Effects of the potent extract on cell viability, TPA-induced colony formation, intracellular reactive oxygen species (ROS) levels, and nuclear factor erythroid 2–related factor 2 (Nrf2)-related protein and mRNA expression, mediated by epigenetic modifications, were evaluated in JB6 P+ cells. Results: Both the water extract (CDL-WE) and the 95% ethanol extract (CDL-95EE) contain abundant flavonoids that inhibit TPA-induced cell transformation and colony formation without minimal cytotoxicity. Mechanistic studies indicated that CDL-95EE increased the gene expression of Nrf2-related detoxification and antioxidant enzymes, such as UDP-glucuronosyltransferase 1A (UGT1A) and heme oxygenase-1 (HO-1), and decreased intracellular ROS accumulation. Furthermore, CDL-95EE reduced the expression of epigenetic modifiers, including DNA methyltransferases (DNMTs) and histone deacetylases (HDACs), suggesting involvement in epigenetic regulation. Conclusions: These findings indicate that CDL, an agricultural by-product, may be useful in cancer prevention through antioxidant and epigenetic mechanisms.

Biomedicines doi: 10.3390/biomedicines14040811

Authors: Amanda Ringwood Chi Zhang Rob Knight

Background: Cutaneous radiation injury arises when ionizing radiation disrupts epidermal barrier integrity, triggering persistent DNA damage, oxidative stress, and senescence-associated inflammatory signaling that drive extracellular matrix degradation and impaired regeneration. Clinical burden is rising due to dose-intensified radiotherapy, but also due to an increased use of energy-based aesthetic procedures that elicit radiation-like dermal injury. Dermal fibroblasts exhibit marked sensitivity to ionizing radiation and rapidly acquire senescence-associated secretory phenotypes that suppress collagen biosynthesis and promote chronic inflammation, underpinning the need for regenerative treatments that restore tissue homeostasis and regenerative competence. Mesenchymal stem cell–derived exosomes have emerged as a promising therapeutic strategy in this setting, with increasing preclinical evidence demonstrating their capacity to attenuate oxidative stress, enhance DNA damage-repair pathways, and normalize fibroblast metabolic function. Methods: In this study, we examine the expression profiles for 14 radiation response–associated genes of irradiated human dermal fibroblasts that were treated with bone marrow and umbilical cord MSC-derived exosomes at different timepoints using quantitative RT-PCR analysis. We also explore functional relationships among these genes through interaction network analysis, and outline a framework to organize pathway-level transcriptional responses to irradiation and exosome treatment. Results: MSC-derived exosome treatment was associated with attenuated early damage response signaling at 24 h, followed by increased expression of genes associated with DNA repair and oxidative stress recovery at intermediate timepoints. Exosome-treated cells also exhibited transcriptional changes consistent with modulation of cell-cycle regulatory pathways and reduced expression of pro-inflammatory markers by 5 d. These findings suggest that MSC-derived exosomes influence the temporal organization of the fibroblast transcriptional response to ionizing radiation and may contribute to molecular programs associated with tissue recovery following ionizing radiation exposure.

Biomedicines doi: 10.3390/biomedicines14040812

Authors: Francesca Nuzzolillo Clarissa Braccia Annapaola Andolfo Stefano de Pretis Michela Palmieri

Background/Objectives: Autophagy is an evolutionarily conserved degradation and recycling pathway through which cells deliver cytoplasmic components, including toxic or damaged proteins and organelles, to lysosomes for clearance. In neurons, which are largely post-mitotic, degradative pathways are essential to prevent the accumulation of cellular waste and to maintain nutrient and energy homeostasis. Increasing evidence suggests that autophagy plays a critical role during early brain development, when neuronal circuits are established, synaptic connections are refined, and activity-dependent mechanisms shape network architecture. However, the developmental regulation of autophagy-related genes and the composition of the autophagic machinery at synapses remain poorly understood. This study aimed to characterize the maturation-dependent dynamics of autophagy–lysosomal genes and to investigate the synaptic autophagy-associated proteome during cortical development. Methods: Genome-wide transcriptomic analyses were performed in the cortical brain region across developmental stages to assess changes in the expression of autophagy–lysosomal genes. In parallel, synaptosomes were isolated and subjected to proteomic analysis to identify autophagy-related proteins associated with synaptic compartments. Results: Transcriptomic profiling revealed stage-dependent regulation of autophagy–lysosomal genes during cortical maturation. Proteomic analysis of synaptosomes identified multiple autophagy-associated proteins enriched at synaptic sites, suggesting that components of the autophagic machinery are present at synapses and may participate in synaptic remodeling and function during key phases of neuronal network formation. Conclusions: These findings provide new insights into the developmental regulation of autophagy in the brain and highlight the potential contribution of synaptic autophagy to neuronal circuit maturation. Understanding these mechanisms may help identify novel therapeutic targets for neurological disorders associated with impaired synaptic and cellular homeostasis.

Biomedicines doi: 10.3390/biomedicines14040810

Authors: Masahiko Itani Tomohiro Aoki

Background: Endovascular treatment has become the major choice for treating intracranial aneurysm (IA). The development of novel endovascular devices for IA treatment is, therefore, socially important. For this purpose, a preclinical animal model to test a prototype of devices plays a crucial role. The major problems regarding currently used preclinical animal models, mainly in medium-to-large animals, are the expense and the lack of IA pathology, as they only mimic the morphological aspect. Methods: Sprague–Dawley rats were used, and the new bifurcation was formed via end-to-side anastomosis of carotid arteries. An aneurysm lesion induced at the newly formed bifurcation site was macroscopically assessed. Endovascular coiling of the induced aneurysm was then done. Results: An aneurysm lesion with a balloon-like shape, as in human cases, was induced at the newly formed bifurcation site. Some of the induced lesions spontaneously ruptured. Endovascular coiling was successfully done by using the micro-catheter and coil used at the clinical site. Conclusions: The rat model of IAs established here provides a novel platform contributing to the development of endovascular devices to treat IAs and, therefore, significantly facilitates the development of devices to achieve more effective treatment.

Biomedicines doi: 10.3390/biomedicines14040809

Authors: Zhendong Cao Wei Liu

Although tremendous progress has been made over the past decades in elucidating the mechanisms of keloid formation and developing therapeutic interventions, keloid remains challenging to cure and prone to recurrence. Historically, keloid has been regarded as a benign skin tumor, with mechanistic investigations and therapeutic efforts focused primarily on keloid tissue and cells themselves. In contrast, Traditional Chinese Medicine (TCM) posits that skin diseases manifest from internal bodily dysfunction or dysregulation. Inspired by the holistic principles of TCM, together with the literature reports and clinical evidence of keloid constitution (a systemic condition characterized by an individual’s predisposition to keloid formation), we propose that keloid is likely a pseudo-skin tumor profoundly influenced by its pathological microenvironment. Accordingly, we propose keloid as a persistent inflammation-driven proliferative skin disorder, and elimination of the inflammatory microenvironment may be essential for curing keloid and preventing relapse. Based on this concept, we developed a holistic therapeutic approach that combines systemic treatment targeting keloid constitution with local therapies, including surgery, chemo/radiotherapy, and compression therapy, for the keloid lesion itself. The systemic component encompasses lifestyle and dietary modifications, stress management, physical exercise, as well as the oral administration of TCM herbal medicines and small chemical compounds to suppress systemic inflammatory and fibrotic status, thereby improving keloid constitution. This article introduces this novel holistic approach along with supportive case studies.

Biomedicines doi: 10.3390/biomedicines14040808

Authors: Ercan Akşit Hasan Bozkurt Mehmet Arslan

Background/Objectives: Acute coronary syndrome (ACS) remains a leading cause of sudden cardiac death worldwide; however, limited data exist regarding the relationship between coronary arterial and coronary venous dynamics in this context. The present study aimed to evaluate whether coronary sinus flow (CSF) dynamics obtained via transthoracic echocardiography (TTE) are associated with the long-term risk of ACS and cardiovascular events. Methods: This retrospective observational cohort study included 100 patients who underwent elective coronary angiography and had their CSF parameters assessed via TTE. All participants were followed up for a duration of up to 48 months. The primary endpoint was cardiovascular mortality, while secondary endpoints comprised the occurrence of ACS, refractory angina pectoris, and cerebrovascular events. Study endpoints were evaluated by dichotomizing the population into two groups using the median CSF value as the cutoff (low: CSF ≤ median; high: CSF > median). Results: The primary endpoint did not differ significantly between the two groups; however, refractory angina was significantly more common in the high CSF group [4 (8.2%) vs. 12 (24.5%), p = 0.029]. Similarly, the high CSF group exhibited a significantly higher rate of the composite endpoint than the low CSF group [13 (26.5%) vs. 25 (51.0%), p = 0.013]. In multivariate Cox analysis, CSF was an independent predictor of the composite endpoint (HR: 1.50; 95% CI: 1.11–2.02, p = 0.009). Conclusions: Baseline CSF measured by TTE was independently associated with the composite endpoint, whereas its association with isolated ACS was limited. CSF assessment may provide incremental prognostic information alongside conventional arterial and microvascular evaluation.

Biomedicines doi: 10.3390/biomedicines14040807

Authors: Jano Mathias Kosing Lucian Mureşan Gabriel Gusetu Radu Rosu Dana Pop Cecilia Lazea Simona Sorana Căinap Alina Negru Gabriel Cismaru

Background/Objectives: The Wolff–Parkinson–White (WPW) pattern is characterized by ventricular preexcitation due to an accessory atrioventricular pathway. Population-based data on the prevalence of asymptomatic WPW patterns in children are limited, and automated ECG interpretation may be misleading in the setting of preexcitation. Our aim was to determine the prevalence of the WPW pattern in a large cohort of asymptomatic Romanian school children and to describe electrocardiographic characteristics, ECG-based accessory pathway localization, and automated ECG interpretation errors. Methods: We performed a retrospective cross-sectional analysis of 12-lead ECGs obtained during a school-based screening program in Romania (May–December 2015). After exclusion of duplicates, technical errors, and participants outside the prespecified age range, 24,112 unique children aged 6–18 years were included. The WPW pattern was adjudicated by pediatric electrophysiologists. Prevalence was estimated using the Wilson score method. Sex differences were assessed using Fisher’s exact test. Results: The WPW pattern was identified in 18/24,112 children, yielding a prevalence of 0.075% (0.75 per 1000). The WPW pattern was more frequent in boys than girls (12/11,858 (0.10%) vs. 6/12,255 (0.048%), p = 0.18). Most cases demonstrated mild preexcitation, with only a minority showing marked QRS widening. ECG-based algorithms suggested a predominance of left-sided accessory pathways. Automated ECG interpretation frequently produced misleading diagnostic statements, including bundle branch block/intraventricular conduction delay (5/18; 27.8%) and pseudo-infarction/ischemia patterns (1/18; 5.6%), and did not explicitly identify WPW/preexcitation. Conclusions: In a large school-based screening cohort of asymptomatic Romanian children, WPW pattern prevalence was 0.074%, with a trend toward male predominance. Most cases exhibited mild preexcitation. Automated ECG interpretation commonly misclassified preexcitation-related ECG findings, highlighting the importance of expert ECG review in pediatric screening programs.

Biomedicines doi: 10.3390/biomedicines14040806

Authors: Boris Dinkov Diana Pendicheva-Duhlenska

GLP-1-based drugs are approved for the treatment of type 2 diabetes mellitus (T2DM) and obesity. Metabolic dysfunction-associated steatotic liver disease (MASLD) affects more than 60% of patients with T2DM, and the gut microbiome plays a critical role in its pathogenesis. The gut–liver axis represents a key mechanistic link between dysbiosis and hepatic steatosis. A narrative literature review was conducted using PubMed, Scopus, and ClinicalTrials.gov (2015–2026). Search terms included “GLP-1 receptor agonist,” “microbiome,” “MASLD,” “MASH,” “NAFLD,” “NASH,” “liraglutide,” “semaglutide,” “tirzepatide,” “dulaglutide,” and “exenatide.” Of 363 identified articles, 330 were excluded due to duplication or non-relevant study design; 33 studies (18 preclinical, 15 clinical) were included. In preclinical models, liraglutide demonstrated normalization of the Firmicutes/Bacteroidetes ratio and increased Bifidobacterium and Lactobacillus spp., while tirzepatide significantly reduced hepatic steatosis and increased Akkermansia abundance in diabetic mice. Semaglutide improved gut barrier integrity, increased Alloprevotella and Alistipes, and ameliorated MASLD in murine models. In clinical studies, tirzepatide achieved MASH resolution in 44–62% of patients in the phase 2 SYNERGY-NASH trial. In August 2025, the FDA approved semaglutide for MASH with fibrosis based on the Phase 3 ESSENCE trial. A recent longitudinal study in T2DM patients showed that baseline microbiome composition predicted glycemic response to semaglutide, without significant changes in microbiome diversity. In conclusion, GLP-1-based therapies demonstrate consistent preclinical associations with gut microbiome modulation and reduction in hepatic steatosis. Baseline microbiome composition has been suggested as a potential predictor of treatment response, supporting a personalized approach to MASLD management and warranting future clinical studies.

Biomedicines doi: 10.3390/biomedicines14040805

Authors: Antonio Sanz-Solas Noelia Pérez-Gómez Jorge Labrador Beatriz Cuevas María Victoria Cuevas Francisco Javier Díaz-Gálvez Gerardo Hermida Rodolfo Álvarez-Nuño Gonzalo Benzo Cristina Alonso-Madrigal María González-Oter Natalia García-Sancha Raquel Vinuesa Andrea Rodríguez-Lopez Jesús Novalbos Natalia Busto Raquel Alcaraz Francisco Abad-Santos Miriam Saiz-Rodríguez

Purpose: Bortezomib (BTZ) is widely used in multiple myeloma (MM), but its toxicity shows marked interindividual variability. This study aimed to identify pharmacogenetic and clinical factors associated with BTZ-related adverse drug reactions (ADRs). Methods: A retrospective and prospective observational study was conducted in 127 MM patients treated with BTZ-based regimens. Polymorphisms in CYP enzymes and ABCB1 were genotyped using qPCR. Associations between genetic variants, treatment response, and ADRs were assessed using univariate and multivariate analyses with Benjamini–Hochberg correction. Results: ADRs occurred in 98.4% of patients, most commonly gastrointestinal toxicity (49%), general toxicity (46%), and peripheral neuropathy (39%). Women showed higher rates of gastrointestinal toxicity and non-peripheral neurotoxicity. Multivariate analysis identified the ABCB1 C1236T A/G genotype as protective against gastrointestinal toxicity, while the CYP3A4 intermediate metabolizer phenotype was associated with increased psychiatric toxicity. TP53 mutations were independently associated with hematologic and renal toxicity. Kaplan–Meier analysis showed earlier onset of peripheral neuropathy and respiratory toxicity in CYP3A4 intermediate and poor metabolizers. Conclusions: Genetic variation in ABCB1 and CYP3A4, together with clinical factors such as TP53 mutation and sex, may contribute to interindividual variability in BTZ safety in MM. These findings should be considered exploratory given the sample size and require confirmation in larger cohorts. Nonetheless, they suggest a potential role for pharmacogenomics in supporting future approaches to treatment personalization.

Biomedicines doi: 10.3390/biomedicines14040803

Authors: Rosa Lauretta Giulia Puliani Marta Bianchini Marilda Mormando Antonietta Fasciglione Maria Flavia Bagaglini Ferdinando Marandino Marialuisa Appetecchia

Background: Solitary fibrous tumors are uncommon fibroblastic neoplasms. These tumors are characterized by the recurrent NAB2-STAT6 gene fusion, which is a hallmark of solitary fibrous tumors (SFTs), particularly those arising in the thoracic cavity. While SFTs are mostly found in the abdomen and pleura, they can occur in various locations, including the head and neck region (6% of cases of SFTs). Solitary fibrous tumors of the thyroid (SFTTs) are extremely rare, accounting for only 0.1% of all thyroid tumors. The gold standard imaging modality for thyroid tumors is ultrasonography, even though distinctive characteristics for these types of neoplasms are absent, making pre-operative diagnosis more challenging. Aim: The aim of this study is to perform a systematic literature review and to describe our case by analyzing the main clinical features, histological diagnostic features and treatments of this rare tumor, in order to clarify the behavior and molecular characteristics of SFTTs. Methods: A comprehensive systematic literature review was conducted according to the PRISMA guidelines for SFTTs. We searched the PubMed and EMBASE databases for articles published up to November 2025. The inclusion criteria include confirmed diagnosis of SFTT, while articles describing unrelated neoplasms or articles that were not in English were excluded. A standardized form was used to extract information on the imaging characteristics, histological diagnosis, treatment and outcome. Results: As of 2025, a total of 43 articles were selected, with 61 reported cases of SFTT in the English literature. Pre-operative diagnosis of SFTT is controversial and usually requires immunohistochemical confirmation. In our case, molecular analysis identified, for the first time, a NAB2ex6–STAT6ex17 fusion, contributing to the molecular characterization of this rare tumor. Conclusions: SFTTs are rare and difficult to diagnose; thus, they require a multidisciplinary approach for accurate diagnosis and management. The combination of imaging, cytology, histopathology, and molecular testing is essential in distinguishing SFTTs from other thyroid malignancies. Surgical excision remains the mainstay of treatment, and long-term follow-up is recommended due to the potential risk of recurrence or metastasis.

Biomedicines doi: 10.3390/biomedicines14040802

Authors: Penghui Chen Jifang Zhang Ying Wang Jiarui Chen

Background: Kölliker’s organ (KO) support cells undergo orderly, time-dependent degeneration that is essential for auditory development and is accompanied by precisely regulated autophagic activity; however, the molecular hierarchy linking autophagy to this remodeling remains obscure. This study aimed to elucidate the regulatory mechanisms connecting autophagic flux to lysosomal biogenesis and auditory function during cochlear development. Method: We established an Atg5flox/flox; Sox2Cre+ mouse model with deletion of the autophagy gene Atg5 in cochlear-supporting cells. Auditory function was assessed via Auditory Brainstem Response (ABR) testing. Transcriptomic profiling of the neonatal basilar membrane was performed to screen for downstream targets. Mechanistic validation included spatiotemporal immunofluorescence mapping (E18–P30) and in vitro functional assays using siRNA-mediated knockdown and lysosomal tracking. Results: At 2 months of age, Atg5flox/flox; Sox2Cre+ mice exhibited moderate-to-severe sensorineural hearing loss accompanied by significant outer hair cell loss. Bulk RNA-seq of the basilar membrane identified fork-head box A3 (Foxa3) as a significantly downregulated transcription factor within the lysosomal–autophagy network. Spatiotemporal immunolabelling from embryonic day 18 to postnatal day 30 revealed that FOXA3 expression becomes progressively restricted to KO cells during postnatal development, with ATG5 loss reducing FOXA3 protein levels by 62.4%. In vitro, deficiency of either Atg5 or Foxa3 in primary KO cells resulted in comparable reductions in LAMP1-positive puncta. Conclusions: These findings support a model wherein the ATG5-FOXA3 axis contributes to lysosomal biogenesis in developing KO cells, with implications for understanding mechanisms of congenital sensorineural hearing loss.

Biomedicines doi: 10.3390/biomedicines14040804

Authors: Mengji Xie Xiaoxuan Ma Ying Zhang Le Kuai Ying Luo Jiankun Song Xiaojie Ding Yi Ru Yue Luo Xiaoya Fei Seokgteong Hong Guoshu Deng Yonghua Su Ruiping Wang Bin Li Yanwei Xiang Miao Li Mi Zhou

Background: Psoriasis frequently relapses after treatment withdrawal, consistent with persistent epigenetic programs in lesional immune cells. Lysine acetylation is a reversible regulatory layer linking chromatin accessibility, transcription factor activity, and immune-cell effector programs; yet, its cell-type-resolved landscape and clinical stratification value in psoriasis remain incompletely defined. Methods: We integrated four bulk transcriptome cohorts of psoriatic and healthy skin (746 psoriasis, 515 controls) with two public skin scRNA-seq datasets. A diagnostic acetylation-regulator signature was derived from 33 curated acetylation regulators, and acetylation endotypes were defined by unsupervised clustering. The cell-type-specific expression was mapped at the single-cell resolution. Key regulators were validated by quantitative real-time polymerase chain reaction (qRT-PCR) in an imiquimod-induced psoriasis-like mouse model, and further verified in an independent dataset (GSE136757). Motif enrichment and drug–target mining were used to prioritize transcriptional regulators and candidate epigenetic therapeutics. Results: Sixteen acetylation regulators were differentially expressed in bulk skin, with histone deacetylase (HDAC1) showing the strongest upregulation and lysine acetyltransferase (KAT2A) the strongest downregulation. A 13-gene acetylation signature discriminated psoriasis from controls (area under the curve, AUC 0.886) and separated lesional samples into two acetylation endotypes with divergent pathway states (hypoxia–glycolysis versus oxidative-stress-dominated programs). Single-cell mapping demonstrated immune-restricted acetylation modules, including CREB binding protein (CREBBP)-enriched neutrophils, histone deacetylase 1 (HDAC1)-high cluster of differentiation (CD)8+ T cells, and lysine acetyltransferase 6A (KAT6A)/lymphoid enhancer binding factor (LEF1)-enriched CD4+ and regulatory T cell (Treg) subsets, coincident with interleukin (IL)-17-related inflammatory programs. In mice, qRT-PCR confirmed the coordinated dysregulation of hub genes and highlighted Hnf1a and Kat6a as reproducible candidates. External validation using the GSE136757 dataset further supports their robust diagnostic performance. Motif analysis nominated interferon regulatory factor (IRF4), YY transcription factor (YY2), and zinc finger protein (ZNF404) as putative transcriptional mediators downstream of acetylation programs, and drug–target mining prioritized epigenetic compounds with subtype-relevant potential, including histone deacetylase (HDAC) inhibitors (e.g., entinostat) and the p300/CREB binding protein (CBP) inhibitor A485. Conclusions: This integrative atlas links acetylation regulators to specific immune compartments, defines acetylation endotypes associated with distinct inflammatory programs, and provides a rationale for stratified epigenetic target selection in psoriasis.