Biomedicines doi: 10.3390/biomedicines14051008

Authors: Theodoros Panou Paschalis Steiropoulos Fotios Drakopanagiotakis

The dipeptidyl peptidase (DPP) family comprises enzymes with important metabolic and immunomodulatory properties. This narrative review summarizes recent clinical and experimental evidence on the role of DPP-1, DPP-4, DPP-9, and DPP-10 in pulmonary diseases. The strongest translational evidence currently supports DPP-1 inhibition in non-cystic fibrosis bronchiectasis, where brensocatib reduces exacerbations and prolongs time to first exacerbation, with additional DPP-1 inhibitors in development. By contrast, the roles of DPP-4, DPP-9, and DPP-10 are supported mainly by preclinical studies in pulmonary hypertension, acute lung injury (ALI)/acute respiratory distress syndrome (ARDS), pulmonary fibrosis, asthma, non-small cell lung cancer (NSCLC), and nonsteroidal anti-inflammatory drugs (NSAIDs)/aspirin-exacerbated respiratory disease. Across these models, DPP inhibition modulates inflammation, protease activation, epithelial or endothelial mesenchymal transition, extracellular matrix (ECM) remodeling, and related signaling pathways. Overall, DPP-targeted interventions are promising in pulmonary medicine, but broader clinical translation will require well-designed prospective trials.

Biomedicines doi: 10.3390/biomedicines14051007

Authors: Lutfi Cagatay Onar Ersin Guner Havva Nur Alparslan Yumun Hasan Dindar Ibrahim Yilmaz Gunduz Yumun

Background: Colchicine is a microtubule-targeting anti-inflammatory agent with emerging relevance in cardiovascular disease; however, its effects on injury-induced vascular remodeling remain incompletely defined. Methods: In this study, a rat iliac artery clamp injury model was used to evaluate the effects of colchicine (0.5 mg/kg/day, oral gavage) over 28 days. Histomorphometric, histopathological, and immunohistochemical analyses were performed to assess vascular remodeling. In parallel, molecular docking and STRING/Cytoscape-based protein–protein interaction (PPI) network analyses were conducted to provide structural and systems-level context. Results: Colchicine significantly reduced intimal thickness, the intima-to-media (I/M) ratio, luminal stenosis, adventitial thickness, and collagen deposition, while preserving the lumen area and improving the remodeling index. Medial thickness was not significantly affected. Proliferative activity showed a decreasing trend without statistical significance. Circulating inflammatory cytokines, including TNF-α and IL-1β, did not differ significantly between groups. Docking analyses suggested potential interactions with β-tubulin, ADAM17, NLRP3, IKKβ, and RELA, while network analysis identified an interaction architecture centered on NF-κB-related regulatory components and inflammasome-associated signaling pathways. Conclusions: Colchicine attenuates injury-induced vascular remodeling in this experimental model. These findings, together with complementary in silico analyses, suggest a multi-target, inflammation-associated framework involving NF-κB-related and inflammasome-linked pathways. The in silico analyses provide supportive mechanistic context but do not establish causal relationships.

Biomedicines doi: 10.3390/biomedicines14051006

Authors: Sıtkı Ün Veli Kaan Aydın Özgür Kurt Gergana Lengerova Martina Bozhkova Steliyan Petrov Aylin Köseler

Background/Objectives: Bladder cancer treatment is frequently hindered by chemoresistance to agents such as cisplatin, a process in which autophagy is hypothesized to play a cytoprotective role. This study aimed to investigate the time-dependent transcriptional dynamics of autophagy-related genes in response to cisplatin in bladder cancer cell lines to better elucidate the molecular underpinnings of this resistance. Methods: Two human bladder cancer cell lines, T24 and 5637, were exposed to varying concentrations of cisplatin. Cell viability and half-maximal inhibitory concentration (IC50) values were determined at 24 and 48 h using the MTS assay. Subsequently, the relative mRNA expression levels of key autophagy-related genes (ULK1, BECN1, ATG5, ATG7, LC3B, SQSTM1/p62, LAMP1, and TFEB) were quantitatively analyzed via RT-qPCR at 0, 6, 24, and 48 h intervals. Results: Cisplatin exerted a dose- and time-dependent cytotoxic effect, with 5637 cells exhibiting significantly greater sensitivity compared to T24 cells. Transcriptional analysis revealed a dynamic, multiphasic modulation of the autophagic pathway: an early-phase upregulation of initiation genes (ULK1, BECN1), a mid-phase increase in autophagosome formation genes (ATG5, ATG7), and a late-phase alteration in lysosomal regulation genes (LAMP1, TFEB). Notably, the more chemoresistant T24 cells mounted a robust and sustained autophagic transcriptional response, whereas the sensitive 5637 cells demonstrated a more limited and transient reaction. Conclusions: Cisplatin modulates the autophagic pathway at the transcriptional level in a highly dynamic, time-dependent, and cell-line-specific manner. Interpreted alongside established functional evidence in the literature, the sustained autophagic gene expression observed in the resistant cells is consistent with a potential cytoprotective role, warranting further functional validation at the protein level. These findings map the temporal genetic landscape of cisplatin-induced autophagy, providing a theoretical framework for optimizing the timing of autophagy-targeted combination therapies in bladder cancer.

Biomedicines doi: 10.3390/biomedicines14051005

Authors: Adelin-Rareș Candrea Laura Ioana Gavrilaș Andrei Mocan Adriana Rusu Gianina Crișan

Background/Objectives: The gut microbiota has been increasingly studied in the context of gastrointestinal, neuropsychiatric, and immune-mediated disorders. However, comparative data across these conditions within the same population remain limited. This observational study aimed to perform a comparative analysis of gut microbiota composition across individuals with irritable bowel syndrome (IBS), anxiety disorders (AG), and autoimmune diseases (AI) and healthy controls (HC) from a Romanian cohort, aiming to identify population-specific microbial patterns. Methods: This study included 59 Romanian adults categorized into IBS (n = 27), AG (n = 13), AI (n = 11), and HC (n = 8) groups. Gut microbiota profiles were analyzed using 16S rRNA gene sequencing (NGS), and fecal short-chain fatty acids were quantified by headspace gas chromatography–mass spectrometry. Results: At the phylum level, Actinobacteria showed significantly higher relative abundance in HC compared to both the AI (adjusted p = 0.028) and the AG (adjusted p = 0.023) groups, with the AG–HC difference confirmed by LinDA analysis (FDR-adjusted p = 0.049). At the genus level, Bifidobacterium spp. was significantly lower in the AG compared to HC (adjusted p = 0.036), while Bacteroides spp. showed higher abundance in the AG compared to AI (adjusted p = 0.013), a finding supported by Kruskal–Wallis analysis. Lactate production was significantly lower in both the AI (p = 0.014) and AG (p = 0.038) compared to HC. The Simpson index revealed significantly lower microbial evenness in the AG compared to IBS and AI (FDR-adjusted p = 0.042). Beta diversity analysis identified significant differences in community composition between groups (PERMANOVA: Pseudo-F = 2.395, R2 = 0.116, p = 0.039), with the most distinct separation between AI and AG (FDR-adjusted p = 0.048). Spearman correlation analysis revealed strong positive associations between Bifidobacterium and lactate, Bacteroides and the acetate/propionate ratio, and Actinobacteria and lactate (all FDR-adjusted p < 0.001). Conclusions: The present study provides exploratory evidence of disease-associated microbial and functional patterns in a Romanian cohort, with reduced Actinobacteria in the anxiety group as the most robust observation, suggesting a potential role of the gut microbiota in gut–brain–immune interactions and contributing preliminary population-specific data from an underrepresented Eastern European population.

Biomedicines doi: 10.3390/biomedicines14051004

Authors: Kai Lin Mei-Wei He Fei Wang Xin-Yu Hu Zi-Yue He Chen-Lu Zhang Zhi-Qiang Huang Hong-Wei Wang

Background/Objectives: Cancer cachexia (CC) is a metabolic syndrome characterized by the progressive loss of skeletal muscle and adipose tissue during tumor progression. Despite its clinical prevalence, effective therapeutic options are currently lacking. Phloretin, a natural flavonoid with potent anti-inflammatory and antioxidant properties, has unclear efficacy against CC. This study investigates the therapeutic potential of phloretin in ameliorating cancer cachexia. Methods: Mouse models of CC were established using BALB/c mice implanted with C26 colon carcinoma cells and C57BL/6 mice implanted with Lewis lung carcinoma (LLC) cells. Upon the detection of palpable tumors, phloretin (10 mg/kg) was administered daily via intraperitoneal injection. At the endpoint, hind limb skeletal muscle, inguinal white adipose tissue (iWAT), and hearts were harvested and weighed. Lean body mass was assessed by analyzing the weight of the carcass following the excision of skin, subcutaneous fat, and visceral organs. Gene expression and protein levels in muscle tissues were subsequently quantified. Results: Phloretin administration significantly alleviated tumor-induced loss of tumor-free body weight. It effectively preserved skeletal muscle mass in both C26 and LLC cachexia models, while significantly attenuating adipose tissue depletion in the C26 model. In vitro, phloretin treatment mitigated myotube atrophy induced by C26 conditioned medium. Mechanistically, phloretin inhibited STAT3 activation in skeletal muscle. This inhibition suppressed the expression of the E3 ubiquitin ligases MuRF-1 and Atrogin-1. Furthermore, phloretin concurrently modulated the autophagy pathway. Conclusions: Phloretin effectively ameliorates cancer cachexia-induced muscle wasting by targeting STAT3-mediated protein degradation and autophagy pathways. These findings suggest that phloretin represents a promising therapeutic agent for the clinical management of cancer-associated cachexia.

Biomedicines doi: 10.3390/biomedicines14051003

Authors: Peihao Yang Demeng Liu Jiyao Wang Chao Liu Yan Fang

Background: Microfibrillar-associated protein 2 (MFAP2) is implicated in various malignancies, yet its specific role and molecular mechanisms in glioblastoma (GBM) progression remain poorly understood. Methods: We analyzed MFAP2 expression in human clinical specimens and murine models. Functional impacts were assessed in U251 cells via gain- and loss-of-function assays. Mechanistic studies explored the interplay between autophagic flux and Wnt/β-catenin signaling. An orthotopic GL261 syngeneic orthotopic model validated these findings in vivo. Results: MFAP2 was significantly overexpressed in GBM, correlating with poor patient prognosis. In vitro, MFAP2 markedly enhanced U251 viability, migration, and invasion while suppressing apoptosis. Mechanistically, MFAP2 triggered autophagic flux, subsequently activating the Wnt/β-catenin cascade and its downstream targets (MMP9, c-Myc, Cyclin D1). Pharmacological inhibition of either autophagy or Wnt signaling effectively abrogated these oncogenic phenotypes. In vivo, MFAP2 knockdown reduced tumor volume by 62.4% and suppressed the autophagy–Wnt axis. Conclusions: MFAP2 is an oncogenic regulator in glioblastoma models that links autophagy activity to Wnt/β-catenin signaling. Our findings support MFAP2 as a candidate prognostic biomarker and a potential therapeutic target; however, additional validation in larger molecularly annotated clinical cohorts and multiple GBM models is warranted.

Biomedicines doi: 10.3390/biomedicines14051002

Authors: Caroline E. R. Souleyrette Phillip C. West Stacy S. Kirkpatrick Joshua D. Arnold Michael R. Buckley Michael B. Freeman Oscar H. Grandas Lauren B. Grimsley Michael M. McNally Deidra J. H. Mountain

Background/Objectives: RNA interference (RNAi) is a promising strategy for mitigating diseases at the molecular level. However, RNAi is limited by its instability in biological fluids and impermeability to cellular membranes. In response, our lab has previously patented a non-ionizable lipid nanoparticle (LNP) platform (R8-PLP) for RNAi therapeutic delivery. This formulation incorporates 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DSPE-PEG) to improve particle stability and drug retention. However, long-anchored PEGylated lipids like DSPE-PEG may impair internalization and stimulate immune responses. The literature suggests substituting short-anchored PEGylated-lipids like 1,2-dimyristoyl-rac-glycero-3-[methoxy(polyethylene glycol)-2000] (DMG-PEG) to attenuate these effects. Here, we evaluated whether substituting DMG-PEG for DSPE-PEG in our R8-PLP would improve in vitro cellular delivery and gene transfection without compromising in vitro critical quality attributes (CQAs) or increasing cytotoxicity. Methods: CQAs [encapsulation efficiency (EE%), particle size (nm), homogeneity (polydispersity index; PDI), and membrane zeta-potential] were assessed at assembly and after storage for up to 28 days at 4 °C. Additionally, in-serum stability at 4 °C and serum release kinetics at 37 °C were assessed. Human aortic smooth muscle cells (HASMCs) were treated with R8-PLPs and analyzed for cellular uptake (fluorometry), cytotoxicity (LIVE/DEAD stain), and gene modulation (qPCR). Results: DMG-PEG incorporation at variable mol% did not alter R8-PLP size, homogeneity, or siRNA EE% at assembly or after long-term storage, but did accelerate siRNA release kinetic profiles compared to DSPE-PEG controls. DMG-PEG substitution enhanced cellular uptake compared to DSPE-PEG R8-PLPs without increasing cytotoxicity. DMG-PEG incorporation also achieved significant silencing versus non-treated controls but did not improve gene silencing compared to DSPE-PEG R8-PLPs. Conclusions: Thus, DMG-PEG substitution did not enhance R8-PLP in vitro gene modulation efficacy despite improving cellular uptake and maintaining CQAs.

Biomedicines doi: 10.3390/biomedicines14051001

Authors: Sri Banerjee W. Sumner Davis Kay Banerjee Joseph McMillan Claret Onukogu Pat Dunn Arturo Olazabal Mekuria Asfaw Heather Esnaola Stephanie Watkins Rafael Gonzales-Lagos

Introduction: The relationship between epilepsy and alcohol use is complex and clinically significant. Alcohol acts as a neurochemical modulator capable of lowering the seizure threshold during both intoxication and withdrawal, while chronic misuse may contribute to epileptogenesis through neuronal injury, metabolic stress, and neurotransmitter dysregulation. However, the long-term impact of alcohol use on mortality among people with epilepsy (PWE) remains insufficiently characterized. The purpose of this study was to assess all-cause mortality risk among individuals with epilepsy based on alcohol use history, stratified by race/ethnicity. Methods: Data from the 2008–2018 National Health Interview Survey (NHIS) were linked to mortality outcomes on 31 December 2019 from the National Death Index (NDI) for U.S. adults aged 18 years and older. PWE and alcohol use were determined using self-reported data. Survival probabilities were estimated using weighted Kaplan–Meier methods, and hazard ratios were calculated using Cox proportional hazards models adjusted for demographic and clinical covariates. Results: Our results indicated that among PWE, alcohol use was associated with increased all-cause mortality. The unadjusted hazard ratio (HR) for alcohol use among individuals with epilepsy was 1.30, increasing to 1.40 after multivariable adjustment. In contrast, alcohol use alone without epilepsy was not associated with elevated mortality risk after adjustment. When stratified by race, the combined effect of epilepsy and alcohol use was significantly associated with increased mortality among Black individuals but not White individuals. Conclusions: In this nationally representative cohort, the combined presence of epilepsy and alcohol use was associated with higher all-cause mortality compared with alcohol use alone. Racial differences were observed, underscoring the need for integrated clinical care and further research into genetic, biological, and social determinants influencing epilepsy outcomes.

Biomedicines doi: 10.3390/biomedicines14051000

Authors: Fatih Saglam Bilal Altunisik Ahmet Yaltir Necip Nas

Background: Substance use disorder (SUD) is associated with systemic inflammatory activation and may influence hematological and coagulation-related pathways. Chronic exposure to psychoactive substances may promote immune activation, endothelial dysfunction, and increased platelet reactivity, thereby contributing to inflammation–coagulation crosstalk. Routine laboratory markers derived from complete blood count and coagulation tests have recently gained attention as potential indicators of these biological alterations. Objective: This study aimed to evaluate hematological inflammatory indices, platelet-related parameters, and coagulation markers in individuals with SUD compared with healthy controls. Methods: This cross-sectional study included individuals diagnosed with SUD (n = 55) and age- and sex-matched healthy controls (n = 40). Hematological parameters, including neutrophil, lymphocyte, monocyte, platelet indices, and derived inflammatory ratios, were analyzed. Coagulation parameters, including prothrombin time (PT), activated partial thromboplastin time (aPTT), international normalized ratio (INR), fibrinogen, and D-dimer levels, were also evaluated. Statistical analyses were performed to compare laboratory findings between groups. Results: Significant alterations were observed in several hematological and fibrinolytic parameters in individuals with SUD. Specifically, the neutrophil-to-lymphocyte ratio (NLR), platelet count (PLT), and D-dimer levels were significantly higher in the study group compared with the controls, suggesting increased inflammatory activity, platelet activation, and fibrinolysis. In contrast, the fibrinogen levels did not show clinically meaningful differences between the groups. Conclusions: Individuals with SUD exhibit alterations in hematological inflammatory indices, platelet-related parameters, and fibrinolytic markers. Routine hematological and coagulation parameters may provide accessible indicators of systemic inflammatory and hemostatic disturbances associated with SUD and may support future research aimed at identifying potential biological markers of addiction.

Biomedicines doi: 10.3390/biomedicines14050999

Authors: Khairul Anwar Abdul Rahman Nabil Mohammad Azmi Shahrun Niza Abdullah Suhaimi Zairul Azwan Mohd Azman Farhana Raduan Khairul Najmi Muhammad Nawawi Shamsul Azhar Shah Geok Chin Tan Yin Ping Wong Sayyidi Hamzi Abdul Raub

Colorectal cancer (CRC) develops gradually from precancerous adenomas and is highly curable when detected early. In Malaysia, however, most cases are diagnosed at advanced stages, leading to poorer outcomes despite the availability of screening programmes such as the immunological Faecal Occult Blood Test (iFOBT). Limited screening uptake and poor adherence contribute to delayed diagnosis. Therefore, effective non-invasive and patient-friendly screening tools are essential. Background/Objectives: This study aims to validate the diagnostic performance of the multi-target stool DNA (mt-sDNA) test. Methods: This cross-sectional validation study was conducted at the Endoscopic Center, Hospital Canselor Tuanku Muhriz (HCTM), from January 2024 to September 2025. Adults aged 18–75 years undergoing elective or emergency colonoscopy were included. Results: Among 246 patients, most were male (56.5%), Malay (65.2%), and aged 55–75 years (91.3%). CRC prevalence was 8.5%. A significant association was observed between age group and colonoscopy findings (p = 0.005), with older individuals more likely to have CRC or adenomatous lesions. Most CRC (90.4%) and advanced adenoma (84.6%) cases occurred in symptomatic patients; however, this difference did not demonstrate a significant association with colonoscopy outcomes (p = 0.069). Per rectal bleeding, constitutional symptoms, altered bowel habit, abdominal pain and constipation were significantly associated with CRC and adenomatous lesions. The mt-sDNA test showed a sensitivity of 63.2%, specificity of 85.0%, positive predictive value of 36.4%, and negative predictive value of 94.4%. Conclusions: Preliminary findings indicate that mt-sDNA demonstrates good specificity and high negative predictive value, but moderate sensitivity and low positive predictive value.

Biomedicines doi: 10.3390/biomedicines14050998

Authors: Bowen Xiao Yong Q. Chen Shaopeng Wang

Background/Objectives: Alzheimer’s disease (AD) remains a progressive neurodegenerative disorder with limited therapeutic options. This study aimed to develop an integrative multi-omics computational pipeline to identify diagnostic biomarkers and prioritize druggable therapeutic targets for AD. Methods: We integrated transcriptomic data from 1047 samples (547 AD, 500 controls) using weighted gene co-expression network analysis (WGCNA) and three machine learning algorithms (LASSO, Random Forest, SVM) with strict separation of training, feature selection, and evaluation. Single-cell RNA sequencing of 48,481 nuclei from entorhinal cortex, two-sample Mendelian randomization (MR) with Bayesian colocalization, and structure-based molecular docking with triplicate 500 ns molecular dynamics (MD) simulations were also employed. Results: Machine learning identified 10 consensus biomarker genes involved in synaptic vesicle cycling, ion transport, and calcium homeostasis (internal test AUC = 0.891, 95% CI: 0.836–0.946; external validation on GSE48350: AUC = 0.847, 95% CI: 0.798–0.896). Covariate-adjusted differential expression and MR with Bayesian colocalization converged on eight immune-related therapeutic targets including APOE, TREM2, and TYROBP (p<0.05; Bonferroni-corrected threshold p<0.00625). Single-cell analysis revealed oligodendrocyte expansion in AD (28.5% versus 24.8%), with target genes predominantly expressed in microglia and astrocytes. Virtual screening of 2634 FDA-approved drugs prioritized 10 exploratory repurposing candidates; indomethacin–TREM2 and celecoxib–CSF1R are primary exploratory candidates given structurally validated binding pockets. Triplicate MD simulations (15 μs aggregate) showed force-field-consistent structural stability (RMSD ≤ 3.2 Å). A quantitative multi-omics convergence framework identified four Tier 1 targets (APOE, TREM2, TYROBP, CX3CR1) supported by ≥5 analytical layers (Pperm=0.0003; note: three of five layers share the same transcriptomic input). Conclusions: These findings provide a multi-evidence computational framework linking diagnostic biomarkers and druggable neuroinflammatory targets for AD. All predictions require experimental validation in biochemical and cellular models before clinical conclusions can be drawn.

Biomedicines doi: 10.3390/biomedicines14050997

Authors: Xiao Chen Zhimin Yin Rui Jiao Hui Han Liangcai Yuan Jie Zhou

Background/Objectives: Diabetic foot ulcers (DFUs) are a common and challenging complication of diabetes, significantly impacting the quality of life for patients due to impaired wound healing. Exploring effective and targeted therapies for DFUs is therefore both important and meaningful. Sulforaphane (SFN), a natural bioactive compound found in cruciferous vegetables, shows promise in this area. However, its role and underlying mechanisms in promoting wound healing in DFUs have not been fully understood. Methods: Human umbilical vein endothelial cells (HUVECs) were cultured under high-glucose conditions to establish an in vitro diabetic model. Cell viability, inflammation, apoptosis, and mitochondrial function were assessed. The expression and activation of Nrf2 were examined following SFN treatment. Additionally, Nrf2 overexpression was performed to validate its role in mediating the protective effects of SFN under high-glucose stress. Results: High-glucose conditions significantly reduced HUVEC viability and increased inflammation, apoptosis, and mitochondrial dysfunction. Treatment with SFN effectively counteracted these detrimental effects. SFN robustly activated Nrf2 signaling, and overexpression of Nrf2 recapitulated the protective effects of SFN, attenuating cellular damage under high-glucose conditions. Conclusions: SFN activates Nrf2 expression and protects HUVECs from high-glucose-induced injury by improving cell viability, mitochondrial function, and inflammatory response. These findings suggest that SFN may serve as a promising targeted therapy for diabetic foot ulcers.

Biomedicines doi: 10.3390/biomedicines14050996

Authors: Lara Klehr Anne Thiele Merle Bendig Christine Kloetzer Thorsten Herr Nursena Armagan Sebastian Strauss Robert Fleischmann

Background/Objectives: Monoclonal antibodies targeting the calcitonin gene-related peptide (CGRP) pathway are effective drugs for migraine prevention. The worsening of symptoms after treatment discontinuation has raised the question of whether these agents are associated with sustained central neurophysiological adaptation. This study investigated treatment-associated changes in sensory processing and cortical network efficiency during preventive treatment with CGRP monoclonal antibodies (mAbs). Methods: Twenty-two patients with episodic migraine (21 female, 46.2 ± 13.8 years) and 22 age- and sex-matched healthy controls underwent visual and somatosensory evoked-potential (VEP, SSEP) assessments and quantitative electroencephalography (qEEG). Patients were investigated before treatment initiation (V0) and after 3 months of CGRP mAb treatment (V3). Healthy controls were assessed once. Results: The lack of habituation of VEPs at V0 shifted toward habituation at V3 following treatment with CGRP mAbs (Δslope: −0.37 ± 0.83, p = 0.03). VEP habituation at V3 no longer differed significantly from controls. SSEP amplitudes remained stable and did not differ significantly between groups across the study interval. Exploratory qEEG parameters indicated a less efficient cortical network organization at V0 that was no longer significantly different from controls at V3. Conclusions: Three months of CGRP mAb treatment was associated with a partial normalization of selected neurophysiological parameters, particularly VEP habituation and exploratory qEEG network measures. Given the study design and small sample size, these findings indicate adaptive changes in multi-domain processing, yet these should not be overinterpreted as proof of disease modification.

Biomedicines doi: 10.3390/biomedicines14050995

Authors: Yujia Lin Panpan Lu Qiang Ding Xiang Tao Qinghai Tan Mei Liu

Opioid-induced constipation (OIC) represents a prevalent adverse effect of opioid analgesics, affecting 60–90% of patients and ssignificantly compromising quality of life. This review delineates the multifactorial pathogenesis of OIC. Peripheral μ-opioid receptor (MOR) activation suppresses enteric neuronal excitability, inhibits intestinal motility and secretion, and impairs rectoanal function. Notably, the colon appears to exhibit a distinctive lack of tolerance to opioids. Enteric glial cell activation has been implicated in neuroinflammation, while interstitial cells of Cajal show impaired pacemaker function. Central mechanisms are increasingly recognized to involve the brain–gut axis. Furthermore, opioid-induced barrier disruption, microbiota dysbiosis, and LPS/TLR4-mediated inflammation are proposed to interact and may contribute to a self-reinforcing cycle. Animal models have been instrumental in dissecting these mechanisms. However, they present limitations in reproducibility, clinical phenotype fidelity, and translational validity, particularly regarding microbiome composition and neuroimmune responses. Future research should prioritize the development of standardized, physiologically relevant animal models incorporating multi-omics approaches, and validate mechanism-based therapeutic strategies, including peripherally acting MOR antagonists and microbiota-targeted interventions, for precision management of OIC.

Biomedicines doi: 10.3390/biomedicines14050994

Authors: Maya Nehemia Hilit Cohen Orly Gruzman Tal Meushar Vega Amador Shimon B. Levy Sorina Grisaru-Granovsky Ofra Ben Menachem-Zidon

Mesenchymal stem/stromal cells (MSCs) are widely recognized as potent modulators of inflammation and immune function in bacterial and viral infections. However, their roles in fungal disease remain comparatively under-defined despite the growing clinical burden of invasive and opportunistic mycoses. This Feature Review synthesizes emerging evidence that MSCs influence antifungal outcomes through two complementary axes: (i) host-directed effects, including modulation of immune responses, particularly macrophage responses, and tissue/barrier conditioning; and (ii) fungus-directed effects (direct antifungal activity mediated by contact-dependent mechanisms and secreted antimicrobial factors). We will summarize how MSCs reshape cytokine and chemokine networks and tune innate immune effector functions, with emphasis on macrophage polarization, pattern-recognition receptor signaling, and downstream phagocytic and fungicidal pathways. In parallel, we will review data suggesting that MSCs can interact more directly with fungal pathogens through sensing, physical engagement, and secretion of antimicrobial mediators while highlighting mechanistic uncertainties and model-dependent limitations. A dedicated section will address MSC-derived secretome products (conditioned media, extracellular vesicles) as a cell-free strategy to enhance antifungal immunity. We will critically evaluate conflicting findings across studies, highlighting that outcomes depend on pathogen and host context. Clarifying these context dependencies is essential to rationally develop MSC or secretome-based interventions that are safe, reproducible, and tailored to specific fungal pathogens and patient populations.

Biomedicines doi: 10.3390/biomedicines14050993

Authors: Qiang Zhang Maoqiang Lin Shishun Yan Fei Huang Haiyu Zhou

Objective: This study aims to analyze the dynamic changes in lymphatic endothelial cell (LEC) markers during the progression of intervertebral disk degeneration (IDD) and to investigate their association with the progression of IDD. Method: In this study, intervertebral disk (IVD) specimens were first collected from patients who underwent open lumbar fusion surgery for spinal fractures (control group, n = 10) and lumbar disk herniation (IDD group, n = 10). Concurrently, a mouse IDD model was established, and IVD specimens were collected from mouse in the Sham group and the IDD group 1, 3, and 6 weeks after modeling (n = 5 per group at each time point). Pathological morphological changes in human and mouse IVD specimens were observed using Hematoxylin and Eosin (H&E) and Masson’s Trichrome staining. The degree of degeneration in the mouse IVD specimens was quantified using a histopathological scoring system. Subsequently, real-time quantitative polymerase chain reaction (RT-qPCR), immunohistochemistry (IHC), and immunofluorescence (IF) staining were employed to examine LEC markers in IVD tissue, including lymphatic vessel endothelial hyaluronan receptor 1 (LYVE-1), podoplanin (PDPN), prospero homeobox protein 1 (PROX-1), and vascular endothelial growth factor receptor 3 (VEGFR-3), as well as matrix metabolism-related markers such as matrix metalloproteinase 13 (MMP-13) and collagen II (Col II). Finally, we performed Spearman’s rank correlation analysis between the histopathological scores of all mouse IVD specimens and the corresponding expression levels of LEC markers. Results: In human IVD tissue, expression levels of LYVE-1, PDPN, PROX-1, and VEGFR-3 were extremely low in the normal group. In contrast, expression of these markers was significantly upregulated in the IDD group. In the mouse IDD model, compared with the Sham group at the same time point, the IDD group exhibited higher histopathological scores in IVD tissue, accompanied by upregulation of LYVE-1, PDPN, PROX-1, and MMP-13, as well as downregulation of Col II. In-depth analysis revealed that these differences between the Sham and IDD groups were not static but exhibited a dynamic pattern of increasing magnitude over time. Concurrently, as the modeling period progressed, the histopathological scores of mouse IVD in the IDD group, as well as the expression levels of LYVE-1, PDPN, PROX-1, and MMP-13, showed a progressive upward trend, while Col II expression progressively decreased. In addition, Spearman’s rank correlation analysis revealed that the expression levels of LYVE-1, PDPN, and PROX-1 in mouse IVD tissue were all significantly positively correlated with histopathological scores. Conclusions: In the process of IDD, the dynamic upregulation of LEC markers is highly consistent with its severity in the time dimension. At the same time, there was also a significant positive correlation between the expression level of LEC markers and the severity of IDD. Taken together, these findings suggest that the dynamic upregulation of LEC markers may be potentially associated with the pathological progression of IDD.

Biomedicines doi: 10.3390/biomedicines14050992

Authors: Elen Deng Manvita Mareboina Ilias Georgakopoulos-Soares Nelson S. Yee

Pancreatic cancer is a leading cause of cancer-related mortality in the United States and worldwide. Most patients are diagnosed with pancreatic cancer at advanced stages, when curative therapy is no longer possible. The stage of pancreatic cancer at diagnosis critically impacts the treatment options and thus the clinical outcomes. Currently, there is no established screening program or tests for its early detection. Studying and understanding how those factors influence the stage of pancreatic cancer at diagnosis helps identify barriers and develop screening strategies. Tumoral and demographic factors, as well as social determinants of health, tend to be associated with localized vs. advanced stage of pancreatic cancer at diagnosis. Socioeconomic factors have been shown to be important mediators of racial disparities in stage at diagnosis as well as germline genetic testing. Recently, screening initiatives, blood-based molecular biomarker tests for early detection of pancreatic cancer, and machine learning-based models for risk prediction and imaging diagnostics have been developed. By determining and understanding the factors associated with the stage at diagnosis, risk-stratified screening can be feasible by combining demographics, genetics, comorbidities, lifestyle, and social determinants. Moreover, regulatory policies that address the social determinants of health can guide the development of screening strategies to allocate resources for equitable access to healthcare and to reduce disparities in patients with pancreatic cancer.

Biomedicines doi: 10.3390/biomedicines14050991

Authors: Yuejun Luo Bo Wang Wanjiang Feng Zibo Xu Hongyu Wu Ziming Yan Haoyu Guo Weifeng Liu

Background: The CD163+ macrophage is considered a key component of the tumor immune microenvironment (TIME) in osteosarcoma (OS) in relation to tumor progression and chemotherapy resistance. However, the relationship between CD163+ macrophage infiltration and the efficacy of neoadjuvant chemotherapy (NACT) in OS remains unexplored. Methods: This study collected a total of 195 biopsy samples from newly diagnosed, pretreated OS patients. The infiltration of CD163+ macrophages was evaluated using immunohistochemical (IHC) staining with anti-CD163 antibody. Additionally, multiplex fluorescence staining (CD8, CD68, CD163, and PDL1) was employed to further characterize the TIME profiles associated with different levels of CD163+ macrophage infiltration. The relationships between various clinical characteristics, survival outcomes, and CD163+ macrophage infiltration levels were also assessed. Results: CD163+ macrophages in biopsy tissues ranged from 2.25 cells/mm2 to 3974.79 cells/mm2 and 1.37 cells/mm2 to 3027.20 cells/mm2 in the training and validation cohorts respectively. Multivariate analysis identified that CD163+ macrophage density was an independent predictor for NACT response. Importantly, patients with high CD163+ macrophage infiltration exhibited poorer DFS, DMFS, and RFS than their counterparts. Conclusions: CD163+ macrophage infiltration was an independent predictor for NACT response. Patients with high CD163+ macrophage density benefited less from NACT and exhibited a more immunosuppressive TIME than low-density patients.

Biomedicines doi: 10.3390/biomedicines14050990

Authors: Raluca Terteşş Lucian Cristian Petcu Constantin Ionescu Ionuţ Bulbuc Anca Daniela Pînzaru Bogdan Florentin Niţu Lavinia-Carmen Daba Elena Mocanu Stela Halichidis Nicolae Cârciumaru Simona Claudia Cambrea

Background/Objectives: Sepsis is defined as life-threatening organ dysfunction caused by a dysregulated host response to infection. Identifying reliable biomarkers that reflect the underlying immune pathophysiology of sepsis and support early risk stratification remains a major clinical priority. This prospective study aimed to evaluate the prognostic value of interleukin-6 (IL-6) measured at ICU admission in patients with sepsis and septic shock. Methods: This prospective observational study included adult patients with sepsis and septic shock admitted to the Intensive Care Unit (ICU) of the Clinical Hospital of Infectious Diseases Constanța between 2021 and 2025. Receiver operating characteristic (ROC) curve analysis with DeLong comparisons, Kaplan–Meier survival analysis, and Cox proportional hazards regression modeling were performed to assess the association between baseline IL-6 levels, in-hospital mortality, and time to death. Results: Among the analyzed biomarkers, IL-6 demonstrated the highest discriminatory performance for in-hospital mortality (AUC = 0.956; 95% CI: 0.893–0.987; p < 0.0001). The optimal cut-off value (>135.14 pg/mL) yielded a sensitivity of 87.65% and specificity of 92.86% (Youden index = 0.805). However, despite this excellent discrimination between survivors and non-survivors, baseline IL-6 levels were not significantly associated with time-to-death in Cox proportional hazards regression analysis. Conclusions: Admission IL-6 showed excellent discriminatory performance for mortality risk stratification but was not associated with survival duration in time-to-event analyses. These findings suggest that IL-6 should be interpreted primarily as an early risk stratification biomarker rather than a predictor of survival duration in patients with sepsis.

Biomedicines doi: 10.3390/biomedicines14050989

Authors: Gabriela Ribeiro Silva Amanda Lewis Rubim Flavia da Cunha Vasconcelos Luciana Bueno Ferreira John Greenman Etel Rodrigues Pereira Gimba

Background/Objectives: Anaplastic thyroid carcinoma (ATC) is one of the most aggressive and lethal forms of malignant neoplasm of the endocrine system, and osteopontin (OPN) has been shown to be aberrantly expressed in this tumor type. Among the five OPN splicing isoforms (OPN-SI), OPN-4 has been recently reported in several tumor types, including ATC, but its functional role(s) have not yet been elucidated. Methods: To characterize OPN-4 roles in ATC cells, OPN-4 was ectopically overexpressed in the c643 ATC cell line, generating the c643/OPN-4 cells. OPN-roles were evaluated by cell functional assays, including cell proliferation and viability, using Carboxyfluorescein Succinimidyl Ester (CFSE), crystal violet, and trypan blue assays. For migration, clonogenicity, cell cycle and apoptosis assays were used. For assessment, c643/OPN-4 cells were cultured in two-dimensional (2D) monolayers or three-dimensional (3D) spheroids with the latter being maintained in a bespoke microfluidic system. Results: OPN-4 overexpression led to a significant reduction in cell proliferation, viability, migration and clonogenicity. c643/OPN-4 cells displayed a significant accumulation in the G0/G1 phase and a decrease in the S phase of the cell cycle; however this did not affect cell death or the expression levels of other OPN-SI. In a spheroid model of c643/OPN-4 cells, no significant differences were found in spheroid size or viability when compared to those formed by control cells. Notably, OPN-4 overexpression enhanced the effects of sorafenib on cell viability under dynamic treatment conditions involving continuous perfusion. Conclusions: These early findings point to the fact that OPN-4 may reduce some aspects of tumor progression features in ATC cells and open new avenues for investigating OPN-4 as a biomarker of therapeutic response in personalized treatment strategies.

Biomedicines doi: 10.3390/biomedicines14050988

Authors: Ivana Milošević Branko Beronja Nada Tomanović Marina Đelić Nikola Mitrović Dragana Kalajanović Ankica Vujović

Background/Objectives: Chronic liver diseases, including metabolic dysfunction-associated steatohepatitis (MASH) and chronic viral hepatitis (CVH), are major global health concerns due to their potential progression to cirrhosis, liver failure, and hepatocellular carcinoma. Because liver biopsy, despite meeting the diagnostic gold standard, is invasive and associated with complications, non-invasive fibrosis assessment tools have been increasingly recommended in clinical practice. This study aimed to compare the diagnostic performance of several non-invasive fibrosis markers (ARR, APRI, FI, FIB-4, API, NFS, BARD) and transient elastography in detecting advanced liver fibrosis (F4) in patients with MASH and CVH. Methods: This retrospective study included 237 adult patients (77 MASH, 160 CVH) who underwent liver biopsy between 2017 and 2025 at the University Clinical Center of Serbia. CVH included chronic hepatitis B (CHB) and C (CHC). Patients were evaluated using serum fibrosis indices and TE, and results were compared to histological staging (F0–F4). ROC analysis assessed diagnostic performance. Results: Cirrhosis (F4) was more common in CVH than MASH (p < 0.001). In MASH, NFS (AUROC 0.931), FIB-4 (0.915), BARD (0.872), and APRI (0.878) showed high diagnostic accuracy for F4. In CHC, APRI (0.931), FIB-4 (0.863), and TE (0.938) had strong performance, while in CHB, TE (0.987) outperformed FIB-4 (0.821). Sensitivity and specificity varied by test and cohort, with TE consistently yielding the best results where available. Conclusions: Non-invasive methods, particularly NFS and FIB-4 for MASH and TE for CVH, effectively identify advanced fibrosis. Their application could significantly reduce the need for biopsy, especially in high-risk groups. TE demonstrated superior accuracy, but access limitations highlight the continued relevance of serum-based scores.

Biomedicines doi: 10.3390/biomedicines14050986

Authors: Yuan Yuan Buyi Zhu Linfei Yang Yumu Leng Feifei Chen Zhenhua Yang Wei Gu Kai Zhang

Background: Lung cancer ranks among the most common and deadly malignant tumors worldwide. Drug resistance is a critical factor hindering the effect of chemotherapy for lung cancer. Exosomes, as intercellular signaling molecule carriers, play an important role in carcinogenesis, metastasis and drug resistance. Our study was aimed at exploring the impact of exosomes derived from docetaxel (DTX)-resistant lung cancer cells on regulating biological behaviors of DTX-sensitive cells, further investigating the molecular mechanisms regarding exosome-mediated intercellular communication. Methods: We extracted and identified the exosomes derived from A549, A549/DTX, H1299 and H1299/DTX cells, and then analyzed the expression of exosomal miR-373-3p between DTX-sensitive and DTX-resistant cells. Cell proliferation and apoptosis experiments were verified using a CCK-8 assay, a colony formation assay, a TUNEL assay and flow cytometry. The molecular interaction between miR-373-3p and PDCD4 was evaluated using a dual-luciferase reporter assay. The function of miR-373-3p was further assessed using an in vivo mouse xenograft model. Results: We found that the exosomal miR-373-3p level from DTX-resistant A549/DTX or H1299/DTX cells significantly exceeded that from DTX-sensitive A549 or H1299 cells. In addition, both exosomes derived from DTX-resistant lung cancer cells and miR-373-3p mimics could promote the proliferation of DTX-sensitive cells and inhibit their apoptosis. Moreover, we identified PDCD4 as a key target gene of miR-373-3p, which could induce the malignant behaviors of DTX-sensitive cells by reducing PDCD4 expression. Conclusions: Our results demonstrated that DTX-resistant lung cancer cells could transfer miR-373-3p to DTX-sensitive cells through exosomes, where miR-373-3p could exert its carcinogenic effect via targeting PDCD4.

Biomedicines doi: 10.3390/biomedicines14050987

Authors: Han Zhou Zhenchao Wu Beibei Liu Yipeng Du Rui Wu Ning Shen

Objective: To identify risk factors for progression to sepsis in patients with non-ventilator hospital-acquired pneumonia (NV-HAP) and to develop a practical nomogram for individualized risk assessment in this population. Methods: We retrospectively screened 408 hospitalized patients with hospital-acquired pneumonia at Peking University Third Hospital between January 2017 and December 2021. After excluding patients with an unclear diagnosis date or missing critical variables required for SOFA score calculation, 368 eligible patients with NV-HAP were included and randomly divided into a training cohort (n = 260) and an internal validation cohort (n = 108). An independent temporal validation cohort of 68 patients admitted between January 2022 and December 2022 at the same center was further used for temporal validation. Univariable and multivariable logistic regression analyses with backward stepwise selection were performed in the training cohort to identify predictors associated with progression to sepsis. A nomogram was then constructed based on the final model and evaluated by discrimination, calibration, and decision curve analysis. Results: A total of 368 patients were included in the model development dataset. The final multivariable model retained six predictors: male sex (OR = 2.393, 95% CI: 1.333–4.296), diabetes (OR = 2.205, 95% CI: 1.126–4.319), coagulation dysfunction (OR = 3.327, 95% CI: 1.726–6.413), PaO2/FiO2 (OR = 0.955 per 10-unit increase, 95% CI: 0.912–1.001), platelet count (OR = 0.900 per 10 × 109/L increase, 95% CI: 0.853–0.949), and bilirubin (OR = 1.176 per 1 μmol/L increase, 95% CI: 1.100–1.258). The nomogram showed acceptable performance, with an apparent C-index of 0.809 and a bootstrap-corrected C-index of 0.792 in the training cohort. The C-index was 0.750 (95% CI: 0.658–0.841) in the internal validation cohort and 0.754 (95% CI: 0.639–0.870) in the temporal validation cohort. Calibration analysis showed acceptable agreement between predicted and observed probabilities, and decision curve analysis indicated a positive net clinical benefit across clinically relevant threshold probabilities. Conclusions: In patients with NV-HAP, male sex, diabetes, coagulation dysfunction, lower PaO2/FiO2, lower platelet count, and higher bilirubin were associated with progression to sepsis. The developed nomogram showed acceptable discrimination, calibration, and clinical utility, and may serve as a practical tool for early individualized risk stratification in patients with NV-HAP.

Biomedicines doi: 10.3390/biomedicines14050985

Authors: Hao Wu Yi Wei Xing-Ding Zhang Lin Qi

Lower-grade gliomas (LGGs) often follow a relatively protracted clinical course; however, a substantial proportion eventually undergo malignant transformation to high-grade, treatment-refractory disease. This process has traditionally been interpreted in the context of stepwise histopathologic progression and recurrent genetic alterations. Increasing evidence, however, suggests that malignant transformation is more accurately understood as an evolutionary process shaped by the interplay among epigenetic constraints, cell-state plasticity, and selective pressures. In this review, we examine current evidence supporting a model in which early LGGs, particularly isocitrate dehydrogenase (IDH)-mutant tumors, are initially maintained in relatively restricted cellular states by metabolically imposed epigenetic programs, but progressively escape these constraints under the cumulative influence of therapy, hypoxia, immune remodeling, and genomic instability. We summarize recent advances demonstrating that progression from lower-grade to high-grade disease is accompanied by cell-state transitions characterized by altered lineage identity, acquisition of stem-like features, increased proliferative capacity, and adaptation to cellular stress. We further discuss how these transitions are reinforced by microenvironmental evolution, including vascular remodeling, extracellular matrix reorganization, and changes in immune composition, thereby creating conditions that favor clonal expansion, invasion, and therapeutic resistance. Particular attention is given to longitudinal, single-cell, and spatially resolved studies, which collectively indicate that malignant transformation is not a discrete event but a continuous process of evolutionary selection and phenotypic reprogramming. Finally, we discuss the translational implications of this framework for early risk stratification, biomarker development, and mechanism-based therapeutic intervention. By reframing malignant transformation in LGGs as a process of cell-state escape under persistent selective pressure, this review aims to provide an integrated view of glioma progression and to highlight new opportunities for precision monitoring and treatment.

Biomedicines doi: 10.3390/biomedicines14050984

Authors: Konstantinos C. Mpakogiannis Paraskevi Chasani Ioanna Nefeli Mastorogianni Konstantinos H. Katsanos Fotios S. Fousekis

Introduction: Despite proven efficacy of anti-TNF agents in inflammatory bowel disease, primary non-response affects up to one-third of patients, while secondary loss of response occurs at 13–21% per patient-year, often requiring dose optimization or switching to alternative advanced therapies. Methods: The present single-center cohort study at the University Hospital of Ioannina included biologic-naïve patients receiving anti-TNF therapy as their first biologic treatment. First anti-TNF treatment failure was defined as discontinuation due to persistent IBD activity despite maximal dose optimization (infliximab 10 mg/kg every 4 weeks, adalimumab 40 mg weekly). Patients with measurable anti-drug antibodies prior to anti-TNF dose intensification or discontinuation were excluded. Of 528 anti-TNF-treated patients, 286 (173 with CD, 113 with UC) met the inclusion criteria and were included in the final statistical analysis. Results: Anti-TNF failure occurred in 32.7% of Crohn’s (CD) and 32.9% of ulcerative colitis (UC) patients. Multivariable Cox regression identified complicated phenotype (stricturing or/and penetrating CD; HR = 1.9, p = 0.032) and concomitant corticosteroid use at anti-TNF initiation (HR = 2.03, p = 0.012) as independent predictors of anti-TNF failure in CD. Age at CD diagnosis showed a trend for statistical significance (HR = 1.02, p = 0.061), and after stratification, age at diagnosis ≥ 40 years conferred higher risk (HR = 1.93, p = 0.016), alongside persistent effects of complicated phenotype (HR = 1.83, p = 0.027) and corticosteroid use (HR = 2.01, p = 0.013). In UC patients, female sex predicted anti-TNF failure (HR = 2.13, p = 0.025). IBD-related bowel resection occurred in 26.6% of patients with CD and in 5.3% of patients with UC. Conclusions: Anti-TNF failure remains common despite optimization. Identifying immunogenicity-independent predictors may enable personalized treatment strategies and improve outcomes.

Biomedicines doi: 10.3390/biomedicines14050983

Authors: Ersin Guner Omer Faruk Yilmaz Muharrem Furkan Yuzbasi Mehmet Albayrak Fatih Ugur Ibrahim Yilmaz

Background: Intervertebral disc degeneration (IVDD) is driven by the interplay between inflammatory signaling, extracellular matrix (ECM) degradation, and impaired cellular adaptation. Although several nutraceutical compounds have been reported to exert protective effects in IVDD-related models, their multitarget mechanisms within integrated molecular networks remain incompletely characterized. Methods: An in silico framework integrating molecular docking with network-based analyses was employed to evaluate resveratrol, quercetin, melatonin, curcumin, and baicalein against a predefined panel of IVDD-associated targets, within an exploratory in silico framework. Binding affinities and interaction profiles were assessed using molecular docking, followed by protein–protein interaction (PPI) network construction, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses, and hub gene identification. Results: Docking analyses revealed binding energies ranging from −4.59 to −13.25 kcal/mol, with curcumin and quercetin showing plausible docking poses across a subset of selected targets under the applied protocol. Network analysis showed a highly interconnected structure centered on key inflammatory regulators, including NFKB1, IL6, TNF, IL1B, STAT3, and NLRP3, together with ECM-associated components such as ACAN, COL2A1, SOX9, MMP13, and ADAMTS5. Enrichment analyses further suggested significant associations with inflammatory signaling pathways, cytokine regulation, and ECM organization. Conclusions: These findings are compatible with a distributed, multitarget interaction pattern of nutraceutical compounds within IVDD-associated molecular networks. By integrating molecular docking with network-based analyses, this study offers a system-level framework for interpreting previously reported effects within a disease-specific context. Docking-derived interaction patterns should be interpreted as qualitative and exploratory observations, as docking scores represent model-dependent estimates and do not establish comparable pharmacological effects across heterogeneous targets. The results should be considered hypothesis-generating and require experimental validation.

Biomedicines doi: 10.3390/biomedicines14050982

Authors: Elena Grossini Teresa Esposito Mohammad Mostafa Ola Pour Carlo Smirne Giovanni Casali Mario Pirisi Vincenzo Cantaluppi Daniele Pierelli Rosanna Vaschetto Sakthipriyan Venkatesan

Cardiac surgery represents a cornerstone of modern cardiovascular medicine, yet it is intrinsically linked to significant systemic stress responses that can compromise remote organ function. Among postoperative complications, cardiac surgery-associated acute kidney injury (CSA-AKI) remains a significant clinical challenge characterized by high morbidity and complex pathophysiology. While hemodynamic instability and ischemia–reperfusion injury are established risk factors, renal dysfunction frequently persists despite optimal perfusion. This observation suggests the involvement of potent circulating mediators in cellular injury. Extracellular vesicles (EVs) are essential for intercellular communication and serve as central hubs for transporting bioactive lipids, proteins, and genetic material. Accumulating evidence indicates that the mechanical and oxidative stress inherent to cardiopulmonary bypass triggers substantial release of EVs from platelets, erythrocytes, and injured vascular tissues. These vesicles may function as vectors that traffic oxidized mitochondrial components and pro-inflammatory cargo to the renal parenchyma. This signaling cascade appears to disrupt renal homeostasis through a proposed “dual-hit” mechanism involving the induction of endothelial dysfunction and endothelial-to-mesenchymal transition (EndMT), followed by tubular epithelial injury via mitochondrial fragmentation, redox imbalance, and downregulation of anti-aging factors. The complexity of these EV-mediated interactions may contribute to an incomplete understanding of why specific patient phenotypes fail to recover. This narrative review examines the mechanisms of surgery-induced EV biogenesis, the molecular pathogenesis of endothelial and tubular damage, and the role of intercellular crosstalk. Additionally, we discuss future perspectives on targeting the “EV vector” through therapeutic apheresis and mitochondrial pharmacotherapy to potentially improve clinical outcomes in high-risk surgical patients.

Biomedicines doi: 10.3390/biomedicines14050980

Authors: Antonio José Bollas Becerra Marcelino Cortés García Jorge Balaguer Germán Carlos Rodríguez-López José María Romero Otero José Antonio Esteban Chapel Luis Nieto Roca Mikel Taibo Urquía Ana María Pello Lázaro José Tuñón

Background: Heart failure with reduced ejection fraction (HFrEF) and chronic kidney disease (CKD) are common in the growing population of elderly patients, yet little evidence specifically targeting this population exists. The purpose of this study is to analyze the effect of SGLT2 inhibition in this cohort. Methods: A single-center, real-world observational study was performed. Patients aged >75 with HFrEF and CKD and theoretical indication for sodium–glucose cotransporter 2 (SGLT2) inhibitors were enrolled. Results: A total of 173 patients were included, with a mean age of 84.7 years, mean left ventricle ejection fraction of 29.5% and estimated glomerular filtration rate of 45.9 mL/min/1.73 m2. During a median follow-up of 39 months, 73 (42.2%) deaths from any cause and 95 (53.3%) major clinical events (composite of mortality and heart failure admission) were recorded. Multivariate Cox proportional hazards regression analyses were performed to identify associated variables, and SGLT2 inhibition showed to be a protective factor for the mortality endpoint (hazard ratio 0.324 [0.117–0.894]). Male sex was shown to be a risk factor for both endpoints, diabetes mellitus for the mortality endpoint and diuretic use for the major clinical event endpoints. Conclusions: In a real-world study, treatment with SGLT2 inhibitors in elderly patients with HFrEF and CKD was associated with a lower rate of all-cause mortality.

Biomedicines doi: 10.3390/biomedicines14050981

Authors: Francesco Biscotti Sara Ragucci Massimo Bortolotti Federica Falà Chiara Perrone Nicola Landi Andrea Bolognesi Antimo Di Maro Letizia Polito

Background/Objectives: Ribosome-inactivating proteins (RIPs) are plant-derived enzymes with potent cytotoxic activity, widely studied as anticancer agents, particularly as toxic payloads in immunoconjugates. Despite numerous encouraging results reported, their clinical application has been limited by their immunogenicity. RIPs from edible plants have been proposed as potentially more suitable candidates due to their possible improved tolerability. However, this aspect still requires validation in vivo in animal models. This study investigated the cytotoxic activity, mechanisms of action and translational potential of sodin 5 (a recently characterized type 1 RIP derived from the edible plant Salsola soda L.) in human colon cancer models, comparing it to the well-known type 1 RIP saporin-S6. Methods: The effects of sodin 5 and saporin-S6 on cell viability, cell death mechanisms and epithelial barrier integrity were assessed on HT29 and Caco-2 cell lines. Sodin 5 cross-reactivity with other anti-type 1 RIP sera was evaluated by ELISA. Finally, its structural characteristics were analyzed. Results: Sodin 5 showed a cytotoxic effect comparable to that of saporin-S6 in HT29 and Caco-2 colon cancer cells, with time- and concentration-dependent reductions in viability. Both type 1 RIPs disrupted the integrity of the intestinal epithelial barrier in mono- and co-culture models and predominantly activated the apoptotic pathway, without inducing necrosis. Sodin 5 exhibited limited immunological cross-reactivity and a conserved catalytic core, supporting its potential relevance as a therapeutic payload for intestinal cancer therapy. Conclusions: Our results indicate that sodin 5 possesses promising characteristics for anticancer applications, particularly in the treatment of intestinal malignancies, where local exposure and repeated administration are often required.

Biomedicines doi: 10.3390/biomedicines14050979

Authors: Tomislav Bulum Martina Tomić

Diabetes is a global pandemic and, with its chronic complications, represents a major health issue worldwide, as 12 [...]

Biomedicines doi: 10.3390/biomedicines14050977

Authors: May M. Alrashed Hajera Tabassum Dara Aldisi Maha H. Alhussain Sadia Arjumand Mahmoud M. A. Abulmeaty

Background/Objectives: Chronic Mild Stress (CMS) provokes neuroendocrine dysregulation and oxidative injury that compromise neuronal integrity and plasticity. Disruption of the canonical Wnt/β-catenin signaling pathway has been increasingly linked to stress-induced neurobiological dysfunction. Vitamin D3, a neuroactive hormone with antioxidant and immunomodulatory properties, may exert neuroprotection through modulation of this pathway and attenuation of oxidative damage. The study aims to investigate whether vitamin D3 mitigates CMS-induced alterations in Wnt/β-catenin signaling, oxidative stress markers, and oxidative DNA damage in male Wistar rats. Methods: Thirty-two male Wistar rats were randomly allocated into four groups (n = 8/group): control, CMS only, CMS + vitamin D3 (1000 IU/kg), and CMS + vitamin D3 (10,000 IU/kg). Vitamin D3 was administered intramuscularly three times weekly for 28 days. Hippocampal mRNA expression of Wnt pathway components and brain-derived neurotrophic factor (BDNF) was quantified by RT-qPCR using the 2−ΔΔCt method. Oxidative stress was evaluated by measuring malondialdehyde, glutathione, superoxide dismutase, and catalase, while DNA damage was assessed via 8-OHdG ELISA. Results: CMS significantly downregulated Wnt1, β-catenin, and Axin2 mRNA expression (p < 0.05) while markedly upregulating GSK-3β (p < 0.001). Expression of BDNF was also reduced (p < 0.05). Biochemically, CMS increased MDA and 8-OHdG levels (both p < 0.001) and decreased glutathione (p < 0.001), superoxide dismutase, and catalase activities (p < 0.05). Vitamin D3 supplementation significantly reversed these transcriptional and biochemical alterations, restoring β-catenin signaling, improving antioxidant defenses, and reducing oxidative and genotoxic damage. Conclusions: Vitamin D3 confers significant neuroprotection under chronic stress by modulating Wnt/β-catenin signaling and attenuating oxidative and DNA damage, thereby enhancing neuronal resilience to prolonged stress exposure.

Biomedicines doi: 10.3390/biomedicines14050978

Authors: Fei Yu Yue Zhu Na Li Qing Peng Fanghang Ye Qianlan Luo Jiajun Xia Xiaoyu Hu

The sodium-taurocholate cotransporting polypeptide (NTCP) plays a critical dual role in liver function: maintaining bile acid (BA) enterohepatic circulation and acting as a receptor for the entry of hepatitis B and D viruses into hepatocytes. This review outlines the impact of various post-translational modifications (PTMs) of NTCP—including phosphorylation, oligomerization, ubiquitination, and glycosylation—on its dynamic regulatory network. These modifications coordinate the modulation of NTCP’s membrane localization, stability, conformational state, and protein interactions, precisely controlling its functions in BA uptake and viral invasion. Targeting this PTM network presents a promising strategy for next-generation therapies that selectively inhibit viral infection while preserving BA transport, overcoming the limitations of conventional inhibitors that indiscriminately disrupt virus–NTCP interactions. By synthesizing recent insights into NTCP PTM research, this article highlights its role as a central regulator of its bifunctional properties and reveals potential avenues for precision therapies in viral hepatitis, cholestasis, and related liver diseases. However, most existing evidence is derived from in vitro or cell-based models, whereas in vivo studies and clinical validation remain limited; thus, the translational feasibility of strategies targeting post-translational modifications of NTCP still requires further investigation.

Biomedicines doi: 10.3390/biomedicines14050975

Authors: Kaicheng Zhou Yanyang Nan Mengyang Li Dongyue Hou Caili Xu Haiyan Yu Jun Feng Dianwen Ju Ziyu Wang

Background: Non-small cell lung cancer (NSCLC) remains a leading cause of cancer-related mortality, and although PD-1/PD-L1 immune checkpoint blockade has improved outcomes in some patients, therapeutic responses remain heterogeneous. Tumor-intrinsic heterogeneity within malignant epithelial populations is increasingly recognized as a critical determinant of disease progression and therapy response. Methods: Here, we constructed a comprehensive single-cell atlas of NSCLC by integrating 650,461 cells from 216 tumor and normal samples. Tumor-derived epithelial cells were reclustered to identify transcriptionally distinct subpopulations. Pseudotime analysis, functional experiments, and in vivo validation using a humanized xenograft model were performed to investigate the role of COL3A1. Results: Reclustering of tumor-derived epithelial cells revealed 25 transcriptionally distinct subpopulations. Among these, a high-risk cluster exhibited coordinated activation of epithelial–mesenchymal transition (EMT) and angiogenesis programs and was associated with poor patient survival. Within this aggressive subpopulation, Collagen type III alpha 1 (COL3A1) emerged as a tumor-intrinsic gene associated with extracellular matrix remodeling and angiogenic signaling. Pseudotime analysis indicated that COL3A1+ cells represent a late-stage, poorly differentiated malignant state. Functional experiments demonstrated that COL3A1 knockdown impaired NSCLC cell proliferation, migration, and invasion. Virtual knockout further suggested that COL3A1 may be associated with transcriptional programs involved in PD-L1 upstream signaling pathways, indicating a potential indirect link between tumor-intrinsic states and immune regulatory networks. Consistently, in vivo silencing of COL3A1 enhanced the antitumor efficacy of PD-L1 blockade. Conclusions: Collectively, our study identifies COL3A1 as a tumor-intrinsic gene enriched in malignant epithelial cells with mesenchymal features and a potential therapeutic target. These findings provide a rationale for exploring combinatorial strategies integrating tumor-intrinsic pathway inhibition with immune checkpoint blockade in NSCLC.

Biomedicines doi: 10.3390/biomedicines14050976

Authors: Stefano Andrea Marchitto Gabriele Abbatecola Rola S. Zeidan Lauren Morgan Riccardo Calvani Anna Picca Mathias Schlögl Matteo Tosato Christiaan Leeuwenburgh Stephen D. Anton Francesco Landi Emanuele Marzetti Stefano Cacciatore

Sarcopenia is an age-related skeletal muscle disorder characterized by reduced muscle mass, strength, and physical performance, as well as increased risk of disability, hospitalization, and mortality. Emerging evidence suggests that gut microbiota alterations may contribute to muscle decline via a microbiota–gut–muscle axis, acting as a context-dependent modulator rather than a primary causal driver. This narrative review synthesizes mechanistic, clinical, and translational evidence linking gut dysbiosis to sarcopenia. Preclinical studies show that microbiota modulation (e.g., antibiotics, probiotics, prebiotics, postbiotics, fecal microbiota transplantation) affects muscle mass, strength, and metabolism through pathways including inflammation, mitochondrial dysfunction, altered short-chain fatty acid production, and impaired anabolic signaling. In humans, observational studies associate lower microbial diversity and reduced short-chain fatty acid-producing taxa with poorer muscle outcomes, but findings are heterogeneous and non-causal. Interventional trials remain limited and characterized by small sample sizes, with effects more consistent for functional outcomes than muscle mass. Overall, the gut microbiota represents a modifiable contributor within the complex biology of sarcopenia. Future studies should integrate microbiome profiling and multi-omics approaches within well-designed clinical trials to identify responder phenotypes and define the role of microbiota-targeted strategies within multimodal interventions.

Biomedicines doi: 10.3390/biomedicines14050974

Authors: Matea Buljubašić Franić Petar Todorović Ivana Tica Sedlar Natalija Filipović Nela Kelam Anita Racetin Andrea Kopilaš Ana Dunatov Huljev Katarina Vukojević

Background/Objectives: Clear cell renal cell carcinoma is the most common subtype of kidney cancer and exhibits marked biological heterogeneity, even among tumors of the same histological grade. Although tumor grade remains a key prognostic parameter, the molecular alterations associated with tumor differentiation are not fully understood. This study aimed to evaluate grade-dependent tissue-level expression patterns of proteins involved in cellular stress response, growth regulation, stemness, and apoptosis in clear cell renal cell carcinoma. Methods: Protein expression of heat shock protein 70, insulin-like growth factor 1, octamer-binding transcription factor 4, and apoptosis-inducing factor were analyzed in human clear cell renal cell carcinoma samples and normal renal cortex. Low-grade and high-grade tumors were compared using immunofluorescence staining combined with semi-quantitative and quantitative image analysis. The proportion of positive signals and the number of positive cells were assessed across tissue compartments. In addition, publicly available transcriptomic data from The Cancer Genome Atlas kidney renal clear cell carcinoma cohort were analyzed to explore associations between gene expression levels and overall survival. Results: Distinct grade-dependent expression patterns were observed for all investigated proteins. Heat shock protein 70, insulin-like growth factor 1, and octamer-binding transcription factor 4 showed a higher expression in normal renal tissue with a progressive reduction across tumor grades. In contrast, apoptosis-inducing factor exhibited increased expression in tumor tissue, particularly in low-grade tumors, with a relative decrease in high-grade carcinomas. Stromal compartments of tumor tissue showed minimal or no expression for most markers. Transcriptomic survival analysis did not reveal significant differences in overall survival between high- and low-expression groups for any of the investigated genes. Grade-stratified transcriptomic analysis of the TCGA KIRC cohort revealed consistent patterns for HSP70 family members and OCT4, with progressive grade-dependent mRNA reduction toward higher grades, while IGF1 showed an inverse mRNA trend and AIFM1 showed a uniform reduction across all tumor grades without a clear inter-grade pattern. Conclusions: The findings demonstrate that stress response, growth-related, stemness-associated, and apoptotic proteins display distinct grade-dependent tissue-level expression patterns in clear cell renal cell carcinoma, with the expression profiles of high-grade tumors being of particular translational interest given the aggressive clinical behavior and therapeutic resistance characteristic of this disease stage. These alterations appear to reflect tumor differentiation and biological behavior rather than independent prognostic value, highlighting the complexity of molecular regulation in renal tumorigenesis.

Biomedicines doi: 10.3390/biomedicines14050973

Authors: Qing Gao Kehong Wei Deshen Pan Yufei Xi Chaoliang Xu Deshui Jia

Background: Small cell lung cancer (SCLC) is an aggressive neuroendocrine tumor characterized by an intrinsic resistance to immunotherapy, primarily due to its low immunogenicity and immune-cold tumor microenvironment. The mechanisms underlying this resistance remain poorly understood. Methods: A systematic screen of 796 epigenetic regulators was performed to identify candidate genes associated with effector CD8+ T cell infiltration and clinical outcomes following chemoimmunotherapy in SCLC. This analysis integrated several public SCLC datasets, including single-cell RNA sequencing (scRNA-seq) data from 20 SCLC tumors, bulk RNA-seq data from the IMpower133 cohort, proteomic profiles from the TU-SCLC cohort, and an independent scRNA-seq dataset of 39 SCLC tumors. In vitro and in vivo functional experiments were conducted to investigate the role of the candidate genes in SCLC. Results: The epigenetic regulator TAF1 emerged as a key candidate, with its expression negatively correlating with effector CD8+ T cell infiltration in SCLC. Clinically, patients with low TAF1 expression in tumors showed better outcomes following atezolizumab-based chemoimmunotherapy, particularly in ASCL1-high tumors. Additionally, TAF1 expression was inversely correlated with MHC-I expression. Knockdown of TAF1 in SCLC cells restored MHC-I expression, suppressed tumor growth in immunocompetent mice, and increased CD8+ T cell infiltration. Conclusions: TAF1 functions as a potential epigenetic suppressor of MHC-I expression in SCLC. Targeting TAF1 may represent a promising therapeutic strategy to enhance immunotherapy efficacy in SCLC.

Biomedicines doi: 10.3390/biomedicines14050972

Authors: Qiuyi Zhang Die Dai Yikun Yang Lihong Guo Jiesheng Su Shiqi Lyu Suni Huang Meng Zhang Jianhua Chang

Background: Small-cell lung cancer (SCLC) represents an aggressive malignancy associated with a poor prognosis, underscoring the critical demand for enhanced monitoring methodologies. Circulating tumor DNA (ctDNA) constitutes a promising non-invasive biomarker; however, reports employing highly sensitive tumor-informed assays in SCLC remain scarce. This investigation aimed to assess the clinical utility of a personalized ctDNA monitoring strategy for predicting therapeutic outcomes and resistance in SCLC patients. Methods: This prospective observational study enrolled patients diagnosed with unresectable SCLC. Whole exome sequencing was conducted on baseline tumor specimens to design customized 16-plex multiplex polymerase chain reaction (PCR) panels. Serial blood samples were obtained at baseline, at six-week intervals during treatment, and upon disease progression. Detection of ctDNA-based minimal residual disease (MRD) was performed using a tumor-informed assay (Huajianwei® bespoke MRD) with ultra-deep sequencing. Results: Among seven evaluable patients, the baseline ctDNA-MRD positivity rate was 100%. A significant positive correlation was observed between the baseline ctDNA levels and radiographic tumor burden (r = 0.821, 95% confidence interval [CI] 0.179–0.973, p = 0.034). Longitudinal analysis indicated that patients exhibiting an early decline in MRD levels demonstrated a non-significant trend toward superior progression-free survival (PFS) compared to those with an MRD increase. Though this between-group difference did not reach conventional statistical significance, it represented a trend-level finding (p = 0.0665, hazard ratio [HR] = 0.24, 95% CI: 0.02–3.19), with no definitive prognostic association confirmed in this pilot cohort. Notably, an elevation in MRD preceded radiographic progression by as much as 135 days in certain instances. Conclusions: This study shows that dynamic tumor-informed ctDNA-based MRD monitoring reflects tumor burden changes and may correlate with clinical outcomes in SCLC, supporting its potential to guide personalized treatment and facilitate earlier therapeutic interventions compared to conventional imaging techniques. Prospective multicenter validation is needed to confirm its clinical utility.

Biomedicines doi: 10.3390/biomedicines14050970

Authors: Beata Krasińska Tomasz Urbanowicz Ievgen Spasenenko Krzysztof J. Filipiak Krzysztof Bartuś Zbigniew Krasiński Andrzej Tykarski Anetta Hanć

Background: Trace elements function as essential micronutrients involved in oxidative balance, mitochondrial activity, and cardiovascular metabolism. Cigarette smoking represents a significant source of toxic metals and may disrupt systemic trace element homeostasis. Alterations in micronutrient and metal balance may contribute to oxidative stress, endothelial dysfunction, and myocardial remodeling, which are central mechanisms in the pathogenesis of heart failure with preserved ejection fraction (HFpEF). This study aimed to investigate whether smokers with HFpEF exhibit distinct hair trace element profiles compared with smokers without HFpEF. Methods: In this prospective pilot study, scalp hair samples were collected from adults undergoing clinical evaluation for suspected cardiovascular disease. Trace element concentrations were determined using inductively coupled plasma mass spectrometry (ICP-MS). Participants were first stratified according to smoking status and subsequently, within the smoker subgroup, according to HFpEF diagnosis based on the Heart Failure Association Pre-test assessment, Echocardiography and natriuretic peptide score (HFA-PEFF) algorithm. Differences in trace element concentrations were analyzed using appropriate statistical tests, with multiple-comparison correction using the Benjamini–Hochberg false discovery rate (FDR). Active smoking was defined as ≥10 cigarettes per day for at least 1 year, and cumulative exposure was quantified in pack-years. Results: Fifty-eight participants were included, including 27 active smokers. In unadjusted analyses, several trace elements differed between smokers with HFpEF and those without HFpEF, including vanadium, lithium, aluminum, and copper. However, after FDR correction, only copper remained significantly elevated in smokers with HFpEF (q = 0.004). Hair copper concentrations were markedly higher in the HFpEF group compared with smokers without HFpEF. These differences were observed alongside echocardiographic features consistent with diastolic dysfunction and structural cardiac remodeling. Conclusions: In this hypothesis-generating pilot study, smokers with HFpEF demonstrated elevated hair copper concentrations, suggesting disturbances in trace element and micronutrient homeostasis. Altered copper metabolism may reflect oxidative stress-related cardiometabolic remodeling associated with HFpEF. These findings raise the hypothesis that cardiometabolic phenotype, rather than smoking exposure alone, may modulate trace element homeostasis in HFpEF; however, causal relationships cannot be established.

Biomedicines doi: 10.3390/biomedicines14050971

Authors: Serin Moon Sooeun Oh Dong-Min Kim Jieun Lee Ahwon Lee

Background/Objectives: MicroRNAs (miRNAs) are key regulators of gene expression and have been implicated in multiple aspects of cancer progression. However, the role of miR-214-3p in breast cancer remains controversial. In this study, we investigated the functional role of miR-214-3p and explored its potential regulatory target in breast cancer, particularly in triple-negative breast cancer (TNBC). Methods: miR-214-3p expression was evaluated in breast cancer cell lines. Luciferase reporter assays were performed to assess functional targeting of the FRK 3′-UTR. Functional assays, including proliferation, migration, and invasion assays, were conducted following miR-214-3p overexpression or FRK silencing. Results: miR-214-3p was markedly upregulated in TNBC cells (MDA-MB-231), while Fyn-related kinase (FRK), a potential tumor suppressor, showed an inverse expression trend. Luciferase reporter assays demonstrated that miR-214-3p functionally targets the 3′-UTR of FRK. Functional analyses revealed that overexpression of miR-214-3p significantly increased cell proliferation, migration, and invasion. Notably, silencing of FRK recapitulated these effects, supporting its role as a functional mediator of miR-214-3p. Conclusions: This study identifies a miR-214–FRK regulatory axis in breast cancer and suggests its contribution to aggressive tumor behavior. Targeting miR-214-3p or modulating FRK activity may represent a potential therapeutic strategy.

Biomedicines doi: 10.3390/biomedicines14050969

Authors: Philippe Pinton

Microbiome-based therapies are reshaping the therapeutic landscape for ulcerative colitis (UC), offering new avenues for disease management beyond conventional immunomodulatory and biologic treatments. UC remains a chronic, relapsing condition with significant unmet clinical needs, as many patients fail to achieve sustained remission or experience adverse effects with current therapies. The gut microbiome has emerged as a central contributor to UC pathogenesis, influencing epithelial barrier integrity, immune homeostasis, and metabolic signaling. Interventions such as fecal microbiota transplantation (FMT) and defined microbial consortia have demonstrated proof-of-concept efficacy in early-phase clinical trials, each leveraging distinct mechanistic strategies. FMT, as a broad ecological intervention, restores microbial diversity and functional redundancy, potentially addressing multiple pathogenic mechanisms simultaneously. In contrast, defined consortia enable precise targeting of specific metabolic and immunological pathways, including short-chain fatty acid production, bile-acid remodeling, epithelial barrier reinforcement, immune modulation, and succinate degradation. Recent clinical evidence suggests that consortia with broader mechanistic coverage may achieve more consistent biological activity than narrowly focused designs. This review synthesizes mechanistic and clinical insights across broad and defined microbial consortia, integrates evidence from randomized controlled trials and early-phase LBP studies, and outlines a precision-medicine framework to guide therapy selection. We highlight the importance of aligning therapeutic mechanisms with patient-specific microbial, metabolic, and immune profiles, and discuss future directions including biomarker-guided stratification, hybrid consortia, and adaptive trial designs. Advancing both broad and defined approaches, while incorporating ecological principles, mechanistic understanding, and patient stratification, will be essential to realizing the full therapeutic potential of microbiome-based therapies in UC.

Biomedicines doi: 10.3390/biomedicines14050966

Authors: Guzel R. Sagitova Anna V. Slizova Andrey O. Morozov Anastasia S. Fatyanova Majid Ebrahimi Warkiani Andrei V. Zvyagin Alexey S. Rzhevskiy

Background: The early detection of prostate and testicular tumors remains challenging as standard diagnostic tools often lack sensitivity and produce ambiguous results. Seminal fluid is a biologically rich medium that closely reflects the state of male reproductive tissues and has therefore emerged as a promising source of non-invasive molecular biomarkers. Objective: This study aimed to critically evaluate the evidence regarding cell-free DNA, RNA, proteins and metabolites in seminal fluid, and to assess their potential for improving the early detection of male reproductive cancers. Methods: A systematic review was performed according to PRISMA guidelines. Comprehensive searches of the PubMed and Scopus databases were conducted to identify original clinical studies analyzing molecular biomarkers in seminal fluid from patients with prostate or testicular tumors. For each study, data were extracted on biomarker types, cohort characteristics, analytical methods and diagnostic performance. Results: Forty-two eligible studies were included, covering multiple biomarker classes. Most were observational, single-center investigations classified as level 3b evidence. Across the different types of biomarkers, seminal fluid was associated with tumor-associated molecular changes. Alterations in the concentration, fragmentation and methylation patterns of cell-free DNA (e.g., GSTP1, RARβ2, LGALS3 and OCT3/4) distinguished malignant from benign conditions with sensitivities of up to 80–100%. RNA-based markers, including microRNAs, small non-coding RNAs, and tRNA fragments, showed improved performance in several studies, with multimarker models achieving areas under the curve (AUCs) of 0.85–0.93. Proteomic analyses identified high-specificity candidates such as TGM4, AMACR, PROS1 and DKK3. Metabolomic profiling further strengthened the diagnostic potential; reduced seminal citrate outperformed prostate-specific antigen (AUC 0.748 vs. 0.548), and reproducible shifts in amino acid and lipid profiles were observed in testicular tumors. However, substantial heterogeneity in study design, patient selection, and analytical platforms was observed. Risk of bias varied, and large prospective validation cohorts were lacking. Conclusions: Current evidence suggests that seminal fluid contains molecular signals associated with tumors that could be used for diagnosis. However, the available data are predominantly exploratory and methodologically heterogeneous. Before seminal fluid-based biomarkers can be considered for routine clinical implementation, robust prospective studies with standardized protocols are required.

Biomedicines doi: 10.3390/biomedicines14050965

Authors: Eva Manuela Pena-Burgos Jose Juan Pozo-Kreilinger Rita María Regojo-Zapata María De La Calle

Background/Objectives: Oocyte donation (OD) pregnancies involve complete maternal–fetal genetic disparity and are associated with increased placental dysfunction and adverse perinatal outcomes. However, a unified histopathological framework to characterize alloimmune-mediated placental injury in OD gestations is lacking. This study evaluates immune and vascular alterations in OD triplet placentas and proposes a structured scoring system, the Placental Allograft Rejection-like Score (PARS), to quantify immunovascular dysregulation. Methods: This retrospective study included all OD triplet pregnancies with placental examination performed during 24 years at a tertiary referral center. Maternal, obstetric, fetal, neonatal, and pathological variables were analyzed at the pregnancy level. Histological and immunohistochemical features previously shown to differ between OD and non-OD pregnancies were grouped into six domains: innate immunity, adaptive immunity, checkpoint regulation, vascular remodeling, complement activation, and trophoblastic behavior. Binary thresholds, immunoreactive scores or established morphological cut-offs, were applied to construct a 20-point score classified into three grades. Results: Forty-five OD triplet pregnancies were analyzed. Intra-pregnancy concordance for PARS components was high, with intraclass correlation coefficient ≥0.70 in 87.3% pregnancies. Increasing PARS grades demonstrated a clear clinical gradient. Grade 3 pregnancies had significantly lower birthweight, higher rates of prematurity (<34 weeks), and increased fetal growth restriction. Placental weight decreased progressively with higher PARS. Histologically, grade 3 placentas showed significantly increased accelerated villous maturation and intervillous fibrin deposition. Conclusions: PARS integrates immune and vascular placental lesions into a structured and reproducible framework that correlates with clinically relevant perinatal outcomes and may support future clinical risk stratification, although further validation in larger, multicenter prospective cohorts is required.

Biomedicines doi: 10.3390/biomedicines14050968

Authors: Timur Mušić Tamara Lah Turnšek

The endocannabinoid system (ECS) is a fundamental regulator of brain and body homeostasis, integrating neural, immune, and stress-related signaling pathways. Dysregulation of ECS components, including cannabinoid receptors (CB1 and CB2), endocannabinoids such as anandamide (AEA) and 2-arachidonoylglycerol (2-AG), and their metabolic enzymes (FAAH and MAGL), has been increasingly implicated in the pathophysiology of neuropsychiatric disorders, including mood, anxiety, psychotic, stress-related, and eating disorders. Altered endocannabinoid signaling contributes to maladaptive stress responses, emotional dysregulation, and impaired synaptic plasticity, highlighting the role of the ECS as a core integrative mechanism. Therapeutic strategies targeting ECS, particularly through FAAH inhibition and the use of plant-derived cannabinoids, such as cannabidiol (CBD), show promise in restoring endogenous homeostasis while minimizing the adverse cognitive and affective effects associated with direct CB1 activation. ECS function and treatment response are further influenced by genetic polymorphisms in CNR1, CNR2, FAAH, and MGLL, as well as epigenetic mechanisms, including DNA methylation, histone modifications, and microRNA regulation. Despite these advances, clinical translation remains limited by interindividual variability, the complexity of ECS interactions, and the relatively small size of existing clinical studies. Future research integrating longitudinal clinical trials with multi-omics approaches is essential to support the development of evidence-based, personalized interventions. Overall, understanding ECS mechanisms and dysregulation provides a valuable framework for the development of targeted therapies in neuropsychiatric disorders.

Biomedicines doi: 10.3390/biomedicines14050967

Authors: Marina Lasa Constanza Contreras-Jurado

Thyroid cancer is the most common malignancy of the endocrine system and represents a biologically heterogeneous disease driven by the interplay between endocrine regulation, oncogenic signaling pathways, and tumor microenvironment dynamics. Although most follicular cell-derived thyroid cancers follow an indolent clinical course, a subset progresses toward aggressive, therapy-refractory phenotypes, underscoring the need for refined molecular understanding and improved biomarkers. This review comprehensively examines the molecular determinants of thyroid cancer progression, with particular emphasis on Thyroid Hormone (TH) signaling, the Mitogen-Activated Protein Kinase (MAPK) and Phosphoinositide 3-Kinase (PI3K)/AKT pathways, and the emerging role of microRNAs (miRNAs). We discuss how oncogenic alterations, most notably the V600EBRAF mutation, act as central drivers of tumor initiation and aggressiveness by sustaining MAPK/ERK signaling, promoting dedifferentiation, metabolic reprogramming, immune evasion, and resistance to targeted therapies. The cooperative role of PI3K/AKT signaling in reinforcing survival, invasion, and treatment resistance is highlighted, emphasizing the network-level integration of oncogenic pathways rather than linear dependency on single drivers. In parallel, thyroid hormones exert context-dependent effects on tumor biology through both genomic actions mediated by nuclear thyroid hormone receptors and non-genomic mechanisms initiated at the integrin αvβ3 receptor, linking endocrine status to cancer progression and therapeutic response. Finally, we review the expanding evidence supporting miRNAs as critical regulators of thyroid carcinogenesis and as promising diagnostic, prognostic, and predictive biomarkers. The clinical validation of miRNA-based panels and circulating miRNAs offers new opportunities to improve preoperative risk stratification, reduce overtreatment, and guide personalized therapeutic strategies. Collectively, these insights support a multidimensional framework for understanding thyroid cancer progression and highlight future directions for precision oncology.

Biomedicines doi: 10.3390/biomedicines14050964

Authors: Thalla Kapre Palakurthi Kolimi Akkireddy Vaskuri Kommineni Sharma Kim Palakurthi

The utilization of immunomodulatory nanomaterials, i.e., leveraging their unique properties to enhance immune responses, represents a transformative approach for the treatment of various diseases. Recent advancements in nanotechnology have enabled the design of nanomaterials capable of delivering immunomodulatory agents in a targeted manner, such as cytokines, antibodies, and nucleic acids, to specific cells or tissues involved in immune regulation. These nanomaterials, including nanoparticles, liposomes, nanogels, nanoemulsions, dendrimers, MXenes and extracellular vesicles, have been increasingly tailored to modulate immune responses with precision and efficacy. This targeted approach not only enhances therapeutic outcomes but also reduces off-target effects, minimizing systemic toxicity. In this review, an overview of immunomodulatory nanomaterials and their biomedical applications are highlighted. Herein, we have discussed different types of nanomaterials and their design strategies, interactions with different immune system components (macrophages, dendritic cells (DCs), neutrophils, T lymphocytes (CD4+ helper T-cells, CD8+ cytotoxic T-cells, regulatory T-cells/Tregs, and memory T-cells), and B lymphocytes), and immunomodulation mechanisms. Furthermore, nanomaterial-based immunomodulation strategies to enhance cancer immunotherapy, wound healing, and bone regeneration and the treatment of infectious diseases, autoimmune diseases, and allergy and are discussed in detail. In addition to therapeutic applications, selected nanomaterial platforms demonstrate significant potential in pharmaceutical formulations by improving drug stability, controlled release, and bioavailability, as well as in cosmetology through skin-targeted delivery, anti-inflammatory activity, immune protection, and enhanced tissue regeneration. Finally, clinical trial updates, challenges and future prospects are outlined. Key findings indicate that lipid-based, polymeric, inorganic nanoparticles and dendrimers provide complementary advantages for immunomodulation, including efficient delivery, controlled release, multifunctionality, and precise immune targeting. Despite safety, regulatory, and scalability challenges, these systems show strong potential for advancing precision and personalized medicine. Taken together, these innovations hold great promise for personalized medicine approaches, wherein nanomaterials can be tailored to individual patient profiles for more effective and precise disease treatment and prevention strategies. This review focuses primarily on the mechanistic interactions between immunomodulatory nanomaterials and immune cells, including macrophages, dendritic cells, neutrophils, T lymphocytes, and B lymphocytes, rather than providing an exhaustive treatment of physicochemical optimization parameters such as particle size or surface modification chemistry, which fall outside the defined scope of this work.

Biomedicines doi: 10.3390/biomedicines14050963

Authors: Tongyu Wang Jiling Zeng Mengquan Tan Meiling Zhong Huixian Zhou Yaling Dai Siyuan Song

Diabetic retinopathy (DR) is a common cause of vision loss in diabetes, and it often progresses without early symptoms. DR reflects injury of the retinal neurovascular unit (NVU), which includes neurons, Müller glia, astrocytes, endothelial cells, pericytes, and immune cells. Chronic hyperglycemia drives oxidative stress, advanced glycation end products–receptor for advanced glycation end products (AGE–RAGE) signaling, mitochondrial injury, and low-grade inflammation. These changes disrupt endothelial junctions, promote leukostasis, weaken pericyte support, increase basement membrane thickening, and lead to capillary dropout and hypoxia. Hypoxia-related signaling increases anti-vascular endothelial growth factor (VEGF) activity, which raises vascular leakage and supports neovascular disease. Glial stress and microglial activation add cytokines and reactive oxygen species, and neural dysfunction can appear early and can weaken neurovascular coupling. Modern diabetes care changes the short-term risk landscape because potent therapies can lower HbA1c quickly. Large and rapid HbA1c reductions can trigger early worsening of diabetic retinopathy (EWDR), mainly in patients with high baseline HbA1c and moderate-to-severe baseline DR. Semaglutide’s retinopathy complication signal in SUSTAIN-6 fits an EWDR-like pattern that tracks with rapid glycemic improvement in vulnerable eyes. In parallel, surgery adds acute stress, inflammation, glucose swings, hemodynamic shifts, and medication interruptions. These factors can worsen microvascular instability during recovery. Current perioperative guidelines and regulatory recommendations describe glucose targets and medication safety considerations, including preoperative interruption of SGLT2 inhibitors to reduce euglycemic ketoacidosis risk; however, the retina-specific implications of these measures remain indirect. This review summarizes current evidence linking NVU biology, EWDR risk, and perioperative diabetes-related factors. It discusses how these factors may interact in patients with diabetes and how they may influence retinal outcomes. The review is intended to synthesize current evidence and mechanistic interpretations rather than to provide formal clinical practice recommendations.

Biomedicines doi: 10.3390/biomedicines14050962

Authors: Lou Marie Salomé Schleicher Dorotea Zivalj Hadid Joseph Farzad Diamee Jan Finderle Antea Krsek Lara Baticic

Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system with onset typically in early adulthood and the potential for long-term disability. Current therapies are initiated after symptom onset and do not address early disease triggers, highlighting the need for preventive strategies. Epstein–Barr virus (EBV) infection has emerged as the strongest candidate upstream factor in MS development. This narrative review provides a focused and critical synthesis of current evidence, with particular emphasis on the prevention perspective and the conceptual framing of MS as a potentially infection-driven disease. We integrate epidemiological, immunological, and mechanistic data while explicitly addressing key uncertainties and limitations in causal interpretation. Longitudinal studies indicate that EBV infection precedes MS onset in most cases and is associated with a markedly increased risk following seroconversion. However, EBV infection alone is not sufficient to cause MS. Proposed mechanisms include immune dysregulation and molecular mimicry, though key uncertainties remain. Based on current evidence, EBV represents a promising but unproven target for MS prevention. Future strategies may include prevention of EBV infection or infectious mononucleosis, alongside improved risk stratification and long-term studies to assess the impact of EBV-targeted interventions on MS incidence.

Biomedicines doi: 10.3390/biomedicines14050960

Authors: Tanja Hochstrasser Leon Kaub Christoph Schmitz

Background/Objectives: Radial extracorporeal shock wave therapy (rESWT) is used to treat neuromuscular disorders such as spasticity, but the mechanisms by which rESWT modulates muscle tone remain incompletely understood. One proposed mechanism involves mechanical perturbation of the neuromuscular junction (NMJ), particularly destabilization of acetylcholine receptor (AChR) clusters in the postsynaptic membrane. Because rapsyn knockout mice are not viable, Caenorhabditis elegans (C. elegans) provides an alternative model for studying neuromuscular signaling, expressing the rapsyn homolog RPY-1, a postsynaptic scaffolding protein involved in AChR organization at the NMJ. This study examined whether loss of RPY-1 alters locomotor responses of C. elegans to radial extracorporeal shock wave (rESW) exposure. Methods: Wild-type worms and rpy-1 knockout worms (rpy-1-KOs) were exposed to defined numbers of rESWs. Locomotor behavior was quantified using automated tracking of parameters describing speed, trajectory and body-wave dynamics. Behavioral responses were analyzed both as absolute values and relative to genotype-specific baseline values. Results: rESW exposure produced pronounced alterations in locomotor behavior across all parameters analyzed. Absolute values revealed baseline differences between genotypes. After normalization to genotype-specific baseline values, wild-type worms and rpy-1-KOs responded similarly to moderate exposure levels. At higher exposure levels, genotype-dependent differences became more apparent. Locomotor impairment was most pronounced immediately after exposure but improved during the subsequent recovery period of three hours. Conclusions: rESWs induced strong but largely reversible locomotor alterations in C. elegans during the first hours after exposure. Loss of the rapsyn homolog RPY-1 modified these responses, particularly at higher exposure levels. These findings indicate that RPY-1 influences behavioral responses to rESW exposure, while direct effects on NMJ structure or AChR organization cannot be determined from the present data.

Biomedicines doi: 10.3390/biomedicines14050961

Authors: Tadashi Shiohama Satoru Takahashi Ryo Takeguchi Yuichi Akaba Hironori Sato Masaki Ishikawa Yusuke Kawashima Asuka Koshi Chihiro Abe Shin Nabatame Keita Tsujimura Hiromichi Hamada Keiichiro Suzuki

Background/Objectives: CDKL5 deficiency disorder (CDD) is a rare X-linked developmental and epileptic encephalopathy characterized by early onset refractory epilepsy and severe neurodevelopmental impairment with autistic features. Despite advances in genetic diagnosis, objective biomarkers reflecting disease mechanisms remain limited. Extracellular vesicles (EVs) circulating in the blood may contain disease-related proteins derived from the central nervous system. This study aimed to characterize the plasma EV proteome in CDD in a hypothesis-generating exploratory framework and identify the candidate molecular pathways associated with this disorder. Methods: Plasma samples from seven patients with genetically confirmed CDD and seven neurotypical developmental controls were analyzed. Extracellular vesicle-containing preparations (EVs-cp) were isolated via immunoprecipitation using antibodies against CD9, CD63, and CD81. Proteomic profiling was performed using data-independent mass spectrometry. Differentially expressed proteins were identified using Welch’s t-test with a false discovery rate correction. Functional enrichment, protein interaction network, and correlation analyses were performed using CDKL5 Clinical Severity Assessment (CCSA) scores. Results: In total, 5617 proteins were identified, of which 3510 were used for quantitative analysis. Compared to the controls, 2108 proteins were upregulated and 158 were downregulated in the CDD samples. Enrichment analysis revealed alterations in vesicle-mediated transport, cytoskeletal organization, and immune-related pathways. Several proteins were also correlated with clinical severity scores. Conclusions: Plasma EV proteomics revealed molecular alterations associated with CDD and provided a potential approach for biomarker discovery and mechanistic investigation.

Biomedicines doi: 10.3390/biomedicines14050959

Authors: Barbara Verro Roberta Oliveri Giovanni Pratelli Marianna Lauricella Diana Di Liberto Anna De Blasio Daniela Carlisi Carmelo Saraniti

Laryngeal squamous cell carcinoma (LSCC) remains a major clinical challenge within head and neck oncology, with five-year survival rates showing minimal improvement over recent decades despite advances in surgical and multimodal therapeutic strategies. Increasing evidence identifies metabolic reprogramming as a central driver of tumor progression, therapeutic resistance, and immune evasion in LSCC. Beyond the classical Warburg effect, LSCC exhibits profound metabolic reprogramming, involving coordinated alterations in carbohydrate, amino acid, lipid, and iron metabolism that support adaptation to hypoxic and nutrient-deprived microenvironments. Hypoxia-inducible factors, particularly HIF-1α, coordinate these key biochemical pathways and enzymatic steps by integrating glycolysis, glutaminolysis, folate-dependent one-carbon pathways, lipid synthesis, and mitochondrial remodeling, while also influencing stromal and immune components of the tumor microenvironment. Metabolic crosstalk between tumor cells, cancer-associated fibroblasts, and immune populations promotes immunosuppression through nutrient competition and accumulation of metabolites such as lactate and lipid-derived mediators. In parallel, dysregulated iron handling and altered ferroptosis susceptibility emerge as key determinants of tumor aggressiveness and treatment response. This review synthesizes current evidence on metabolic rewiring in laryngeal squamous cell carcinoma, highlighting how alterations in metabolic pathways create targetable vulnerabilities that drive tumor biology, immune modulation, and resistance to conventional and emerging therapies. Elucidating these metabolic dependencies may support the development of metabolism-based biomarkers and therapeutic strategies in laryngeal squamous cell carcinoma, providing an integrated and translational perspective that links tumor metabolism with microenvironmental interactions and immune modulation, while highlights emerging therapeutic vulnerabilities.

Biomedicines doi: 10.3390/biomedicines14050958

Authors: Sabina E. Nagieva Svetlana A. Smirnikhina

Background/Objectives: TCIRG1-associated infantile osteopetrosis is a severe hereditary disorder caused by impaired osteoclast function, leading to osteosclerosis, hematological abnormalities, neurological complications, and early mortality. Early diagnosis and intervention are critical. Methods: A literature-based analysis was performed on clinical manifestations, outcomes of allogeneic hematopoietic stem cell transplantation (HSCT), immunomodulatory therapy, and experimental gene therapy and cell-based approaches, including lentiviral vectors and patient-derived induced pluripotent stem cells (iPSCs). Results: Allogeneic HSCT is the only established curative therapy, restoring osteoclast function and preventing severe complications. Early transplantation with HLA-matched donors and myeloablative conditioning provides optimal outcomes. Interferon γ1b can transiently enhance osteoclast activity but is not curative and shows variable efficacy. Preclinical studies demonstrate that lentiviral TCIRG1 delivery and transgenic correction in patient-derived iPSCs restore osteoclast function and bone resorption, with stable gene expression and minimal toxicity. Base and prime editing approaches offer potential for precise correction of single-nucleotide TCIRG1 variants, minimizing risks associated with double-strand DNA breaks. Conclusions: Allogeneic HSCT remains the standard therapy for TCIRG1-associated infantile osteopetrosis. Gene therapy and cell-based strategies represent promising adjuncts or alternatives, potentially avoiding immune-related complications and expanding therapeutic options. Further studies are needed to ensure safety, stable engraftment, and long-term efficacy, supporting translation of gene therapy into clinical practice.

Biomedicines doi: 10.3390/biomedicines14050956

Authors: Fan Xiao Zhilu Lei Bo Wu Zhenyu Niu Guifang Deng Linjing Su Yaqian Cao Kerong Qi Xiaoqing Sun Qike Tan Junyu Ke Yanwu Li

Background: Colorectal intraepithelial neoplasia (CR-EN) is a precursor lesion of colitis-associated colorectal cancer (CAC). This study investigated the interventional effects and molecular mechanisms of Rhapontici Radix extract on CR-EN. Methods: An azoxymethane/dextran sulfate sodium (AOM/DSS)-induced mouse model of colonic intraepithelial neoplasia, bioinformatics analysis, organoid models, and HCT116 cell experiments were employed, coupled with histopathological examination, inflammatory cytokine detection, Western blot, immunofluorescence, and HPLC-MS/MS. Results: The results showed that the YAP/AKT-PI3K signaling pathway is aberrantly activated in CRC. Rhapontici Radix extract ameliorated colonic pathology, suppressed inflammatory responses, and remodeled gut microbiota composition in model mice. The extract selectively inhibited the proliferation of CR-EN organoids by downregulating Ki67 and β-catenin while upregulating p53, and suppressed the proliferation, colony formation, and migration of HCT116 cells. Mechanistically, the extract modulated the YAP/PI3K/AKT pathway by upregulating phosphorylated YAP (p-YAP) and downregulating phosphorylated AKT (p-AKT), phosphorylated PI3K (p-PI3K), and their downstream targets p-SRC and c-MYC. Conclusions: This study suggests that Rhapontici Radix extract intervenes in inflammation-associated carcinogenesis through a multi-pathway, multi-target strategy, offering potential therapeutic targets for CAC prevention and treatment.

Biomedicines doi: 10.3390/biomedicines14050957

Authors: Yeongji Jang Hyun Kyu Song Man Kyu Shim Yoosoo Yang

Lipid nanoparticles (LNPs) have become an important platform for the delivery of RNA therapeutics, including messenger RNA (mRNA) and small interfering RNA (siRNA). However, most clinically approved LNP formulations exhibit strong liver tropism following systemic administration, which limits efficient delivery to extrahepatic tissues. This inherent biodistribution profile has therefore been recognized as a key challenge for expanding the therapeutic applications of RNA nanomedicine. Recent efforts have focused on engineering functionalized LNP systems to improve delivery specificity beyond the liver. Surface modification with targeting ligands—such as antibodies, peptides, and nucleic acid aptamers—can promote receptor-mediated uptake by specific immune cell populations, including macrophages, dendritic cells and T lymphocytes. In parallel, advances in lipid design have improved intracellular RNA delivery by facilitating endosomal escape. These developments have broadened the potential use of RNA nanomedicine for inflammatory disorders, including autoimmune diseases, neuroinflammation, and cardiovascular inflammation. Functionalized LNPs are also being investigated for in vivo engineering of immune cells. This review summarizes current strategies for designing functionalized LNP systems, highlights their emerging applications in immune and inflammatory diseases, and discusses key challenges for clinical translation.

Biomedicines doi: 10.3390/biomedicines14050955

Authors: Alissa Groenendijk Bruce J. Aronow Nicholas Cost Mariana Cajaiba Lindsay A. Renfro Elizabeth J. Perlman Lisa Dyer Teresa A. Smolarek Elizabeth A. Mullen Sameed Pervaiz Somak Roy Phillip J. Dexheimer Peixin Lu Peter F. Ehrlich M. M. van den Heuvel-Eibrink Jeffrey S. Dome James I. Geller on behalf of the COG Renal Tumor Committee on behalf of the COG Renal Tumor Committee

Background: Translocation morphology renal cell carcinoma (tRCC) accounts for nearly half of all pediatric RCC cases. Biological study AREN14B4-Q aimed to characterize the molecular landscape of tRCC using samples acquired from patients enrolled in the Children’s Oncology Group Risk Classification and Biobanking study AREN03B2. Methods: From 2006 to 2014, patients <30 yr old with renal tumors were prospectively enrolled in AREN03B2, a Central IRB-approved biobanking study. All pediatric RCC cases underwent a detailed central pathology review and molecular diagnostics to accurately classify RCC subtypes. Samples with confirmed tRCC and appropriate informed consent were identified with adequate tissue for RNA and DNA extraction, along with germline DNA, for whole-genome sequencing (WGS), RNA sequencing, and DNA methylation analyses. Results: From 41 patients, high-quality samples allowed for 18 tumors and non-tumor DNA to be analyzed via WGS, 19 via DNA methylation, and 36 RNA samples via transcriptome sequencing. Consistent with and extending clinical cytogenetic findings, WGS and fusion transcript analyses confirmed very few additional mutations beyond the tRCC translocation. No recurrent genomic copy number gains/losses were found. RNA and WGS analyses enabled sub-classification of tRCC, closely aligning with the different TFE3 fusion partners. DNA methylation analyses demonstrated less tRCC sub-stratification compared with RNA analyses. Pathways activated in tRCC were involved in epithelial differentiation, extracellular matrix organization, apoptosis, immune regulation, signal transduction, and angiogenesis. Conclusions: Arrested epithelial differentiation is the overarching driver in tRCC and is strongly correlated with the specific subclasses of fusion transcript generated by the genetic translocation TFE fusion partner. Negative regulation of apoptosis, increased M2 macrophage expression, and enhanced angiogenesis also appear to be functional features of tRCCs, as are increased expression of matrix metalloproteinases, PI3K-AKT/mTOR/MAPK signaling, and mitochondrial metabolism, highlighting potential therapeutic options beyond direct targeting of the oncogenic driver fusions.

Biomedicines doi: 10.3390/biomedicines14050954

Authors: Thais Zielke Heidi G. Sutherland Neven Maksemous Robert A. Smith Lyn R. Griffiths

Background: Familial hemiplegic migraine (FHM) is a rare and severe form of migraine disorder featuring aura symptoms that include hemiplegia during attacks. While pathogenic missense variants in CACNA1A, ATP1A2, and SCN1A can cause FHM or its sporadic form, they explain less than 20% of suspected hemiplegic migraine cases, suggesting the involvement of other genes or genetic variations, potentially including copy number variations (CNVs). PPRT2 gene variants including CNVs have also been implicated in hemiplegic migraine. Methods: Multiplex ligation-dependent probe amplification (MLPA) assays were used to investigate the presence of CNVs in the CACNA1A, SCN1A, ATP1A2, and PRRT2 genes in a cohort of 170 unrelated probands suspected to have FHM who had tested negative for pathogenic missense or small indel variants within these genes. Potential CNVs were subsequently confirmed using quantitative PCR. Results: In 15 patients referred for FHM genetic testing, various CNVs in the target genes were detected by MLPA and subsequently validated by quantitative PCR. CACNA1A exon duplications were identified in six patients and deletions found in two. Two patients had ATP1A2 exon deletions, while one had a duplication. For SCN1A, exon deletions were found in three patients and a duplication in one. PRRT2 exon deletions were detected in five patients, with a single nucleotide polymorphism (SNP) array confirming a deletion spanning PRRT2 and neighbouring loci including 26 genes in one of those. Three patients had CNVs in more than one FHM gene. Conclusions: Our study demonstrates the presence of CNVs in FHM genes in a subset of hemiplegic migraine cases (~9%), suggesting a likely role in the disorder and highlighting the need to explore structural variation in addition to the commonly interrogated genetic mutation points. These findings contribute to further understanding of genetic mechanisms that underlie hemiplegic migraine and may inform improved diagnostic and therapeutic strategies.

Biomedicines doi: 10.3390/biomedicines14050953

Authors: Soyoung Kwak Jin Kyung Kim Yong-Uk Lee Hye Suk Baek Ye Jin Kwon Mee-Na Park Jeong-Ho Hong Seung-Bo Lee Hae Won Kim Shin Kim

Alzheimer’s disease (AD) is a progressive neurodegenerative disorder in which amyloid β accumulation, tau pathology, chronic neuroinflammation, and metabolic stress converge to drive synaptic dysfunction and neuronal loss. Rather than resulting from a single mechanism, increasing evidence indicates that neurodegeneration in AD is mediated by the coordinated activation of multiple regulated cell death pathways. These pathways include apoptosis, necroptosis, pyroptosis, ferroptosis, and autophagy-dependent cell death, each characterized by distinct molecular mediators and execution programs. Evidence from human brain tissues, animal models, and in vitro systems demonstrates that core pathological drivers such as amyloid β and tau pathology, oxidative stress, and sustained neuroinflammation engage these death pathways in a spatially, temporally, and cell-type-dependent manner across neurons and glial populations. In this review, we synthesize the current knowledge on regulated cell death mechanisms in AD, emphasizing their molecular signatures, cellular specificity, and stage-dependent involvement, together with recent advances in immunohistochemical, imaging, and biofluid-based approaches for detecting neuronal death. By integrating evidence across molecular, cellular, and system levels, this review positions regulated cell death as a unifying framework for understanding neurodegeneration and developing pathway-specific biomarkers and combinatorial neuroprotective strategies.

Biomedicines doi: 10.3390/biomedicines14050952

Authors: Jan Mateusz Zaucha Jan Maciej Zaucha Agnieszka Piekarska

Background: Chimerism analysis is a key tool for monitoring donor-cell engraftment and the risk of relapse and graft-versus-host disease (GVHD) following allogeneic hematopoietic cell transplantation (allo-HCT). The advantage of lineage-specific chimerism assessment, and its dynamics in patients receiving post-transplant cyclophosphamide (PTCy)-based GVHD prophylaxis, remains unclear. Objective: This review summarizes the current state of the art on chimerism analysis in patients with myeloid neoplasms undergoing allo-HCT with PTCy, with emphasis on lineage-specific testing and modern methodologies. Methods: A structured literature review was conducted to assess chimerism dynamics in whole blood (WB), bone marrow, and peripheral blood (PB) subpopulations, including T-cells, CD34+, myeloid, B, and NK (natural killer) cells, and their association with clinical outcomes following PTCy. Results: Lineage-specific PB chimerism, particularly in T-cells, myeloid lineage and CD34+ cells, is more sensitive than WB chimerism for predicting relapse. Declining donor myeloid chimerism or persistent myeloid mixed donor chimerism (MDC) may precede hematologic relapse and provide an early signal of graft instability or ineffective graft-versus-leukemia activity. T-cell MDC has been associated with an increased risk of relapse and a lower risk of GVHD, although persistent T-cell MDC in some patients may instead indicate immune tolerance. Declining CD34+ donor chimerism correlates with a higher risk of relapse and inferior survival outcomes and may therefore complement measurable residual disease testing. Data regarding B-cell and NK-cell chimerism remain inconsistent, likely influenced by delayed immune reconstitution. Compared to anti-thymocyte globulin, PTCy may promote higher donor T-cell chimerism, though findings across studies are variable. Next-generation sequencing (NGS) enables more sensitive detection of microchimerism and relapse prediction. Conclusions: Chimerism analysis, particularly when lineage-specific and NGS-based, offers valuable prognostic insight in allo-HCT with PTCy. Further prospective studies are needed to standardize testing and guide personalized post-HCT strategies.

Biomedicines doi: 10.3390/biomedicines14050951

Authors: Chengjing Shen Taierpuke Maimaiti Gulijikere Kuerban Mireguli Abulimiti Jialu Hu Zilala Yalihong Aikebaier Maimaiti

Background: Lung cancer remains the leading cause of cancer-related mortality worldwide, with non-small cell lung cancer (NSCLC) accounting for approximately 85% of all cases. Isorhamnetin (ISO), a natural dietary flavonoid, has demonstrated potent anti-lung cancer activity in cell models. However, its precise mechanism of action within the complex landscape of NSCLC remains to be fully elucidated. Methods: The effects of ISO on NSCLC cell viability, apoptosis, and cell cycle distribution were assessed in A549 and H1650 cells using the MTT assay, Annexin V-FITC/PI staining, and flow cytometry. Wound healing and Transwell assays were employed to evaluate the isorhamnetin impact on cell migration, invasion, and adhesion. To investigate the underlying molecular mechanisms, RNA sequencing (RNA-seq) was performed, followed by validation of key target genes and proteins using qRT-PCR and Western blot analysis. Results: ISO treatment elicited a significant, dose- and time-dependent inhibition of NSCLC cell viability, which coincided with a marked induction of apoptosis. Cell cycle analysis revealed that ISO triggered an S-phase arrest. Transcriptomic profiling identified ELFN1 and TMEM186 as significantly upregulated genes, while SETDB1 was downregulated in a concentration-dependent manner; this was accompanied by a concomitant upregulation of FGFBP1 protein expression. Functionally, ISO effectively suppressed the migratory, invasive, and adhesive capabilities of both cell lines. Conclusions: Our findings demonstrate that ISO exerts a potent anti-proliferative and anti-metastatic effect on NSCLC cells. The underlying mechanism is multifaceted, involving the induction of apoptosis and cell cycle arrest, coupled with the modulation of a novel regulatory network centered on ELFN1, TMEM186, SETDB1, and FGFBP1. These results provide new mechanistic insights into the anti-tumor pharmacology of isorhamnetin and highlight its potential as a therapeutic agent targeting both cancer cells and their supporting microenvironments.

Biomedicines doi: 10.3390/biomedicines14050950

Authors: Wen Zhang

Genetic research on neuropsychiatric disorders and other complex diseases is advancing rapidly [...]

Biomedicines doi: 10.3390/biomedicines14050949

Authors: Bryan Correa Gonzalez Akshaya Karthikeyan Love A. Moore Anamitra Bhaumik Ethan Sandoval Marion Hardy John D. McPherson Hong Li Mamta Parikh Marc Dall’Era Allen C. Gao Alan P. Lombard

Background/Objectives: Combinations of PARP inhibitors (PARPi) and androgen receptor pathway inhibitors (ARPi) have led to clinical success in treating advanced prostate cancer. However, it is unclear where in the clinical paradigm these combinations will fare best, and their mechanism of action remains unclear. We sought to address open questions and explore alternative strategies to enhance PARPi efficacy. Methods: Viability and morphology were assessed in response to (1) abiraterone, olaparib, or combination and (2) enzalutamide, talazoparib, or combination in castration-resistant C4-2B cells and abiraterone- or enzalutamide-resistant derivative cell models (ARPi-resistant). The efficacy of the ATM inhibitor lartesertib with and without a PARPi was also determined. Western blots and RNA-sequencing were used to interrogate the mechanistic effects of treatment. Results: PARPi and ARPi combinations were effective in all models but provided the most benefit in ARPi-sensitive C4-2B cells. Mechanistically, ARPi was not found to affect homologous recombination repair gene expression but may increase PARP activity. Prolonged PARP inhibition was found to increase the expression of AR target genes, and PARPi pre-treatment increased sensitivity to enzalutamide. ATM inhibition significantly increases PARPi efficacy and appears to outperform ARPi-containing combinations in ARPi-resistant models. Conclusions: PARPi and ARPi combinations are effective in ARPi-resistant models, but efficacy appears stronger in ARPi-sensitive CRPC cells. Presented findings support a novel hypothesis that PARP inhibition may increase ARPi sensitivity with increasing AR activity. Additionally, ATM inhibition may provide more benefit than an ARPi in combination with a PARPi in ARPi-resistant settings. These findings support continued PARPi development for improving patient outcomes.

Biomedicines doi: 10.3390/biomedicines14040948

Authors: Asbiel Felipe Garibaldi-Ríos Luis E. Figuera Belinda Claudia Gómez-Meda Guillermo Moisés Zúñiga-González Ingrid Patricia Dávalos-Rodríguez Patricia Montserrat García-Verdín Ana María Puebla-Pérez Irving Alejandro Carrillo-Dávila Martha Patricia Gallegos-Arreola

Background: Breast cancer (BC) is a leading cause of cancer-related mortality worldwide and a major public health concern in Mexico. Regulatory variants in KRAS, particularly within the 3′UTR and intronic regions, may influence gene expression through microRNA binding and transcriptional regulation. Methods: Five regulatory single-nucleotide variants (SNVs) in KRAS (rs12228277, rs1137196, rs8720, rs12587, and rs12245) were genotyped in BC patients and cancer-free controls. Associations were evaluated using odds ratios (ORs) with 95% confidence intervals (CIs), adjusting for age, alcohol, and tobacco use. Multiple testing was corrected using the Benjamini–Hochberg false discovery rate (FDR). Linkage disequilibrium (LD), multilocus combinations, and in silico functional analyses were also performed. Results: Variants rs12228277, rs1137196, rs8720, and rs12245 showed significant genotype-level associations with BC susceptibility, all remaining significant after FDR correction (pFDR < 0.05). No clinicopathological associations remained significant after correction in single-variant analyses. Multilocus analysis identified specific high-risk combinations (e.g., involving rs12228277, rs1137196, and rs8720) associated with increased BC susceptibility. At the nominal level, these combinations showed associations with clinicopathological features, including hormone receptor–positive status (PR and ER), proliferation markers, and Luminal B subtype; however, none remained significant after FDR correction. LD analysis indicated weak linkage among variants. In silico analyses suggested potential regulatory effects on microRNA binding and KRAS expression. Conclusions: Regulatory variants in KRAS are associated with BC susceptibility through independent effects and potential combinatorial patterns. These findings support the relevance of non-coding variation in cancer risk and warrant further functional and replication studies.

Biomedicines doi: 10.3390/biomedicines14040947

Authors: Emilia Daliana Muntean Daniela-Cornelia Lazăr Ana-Maria Pah Christian Banciu Sorin-Dan Chiriac Iasmina Denisa Boantă Florin Muntean Iulian-Alexandru Blidişel George-Andrei Drăghici Radu Jipa

Gastric cancer remains a highly heterogeneous malignancy in which chemotherapy response is limited by intrinsic and acquired resistance, cumulative toxicity, and the restricted predictive value of conventional preclinical models. This review critically synthesizes evidence on selected medicinal plants and their bioactive phytocompounds as adjuncts to standard chemotherapy for gastric cancer, with an emphasis on mechanistic plausibility, preclinical synergy, and translational barriers. Across the reviewed literature, phytocompounds from Curcuma longa, Scutellaria baicalensis, Camellia sinensis, Syzygium aromaticum, Glycyrrhiza glabra, Allium sativum, Marsdenia tenacissima, and Rhus verniciflua showed anticancer or chemopreventive activity through multitarget effects on apoptosis, proliferation, invasion, inflammation, oxidative stress, and resistance-associated signaling. The most convincing chemosensitizing evidence involved curcumin, wogonin, baicalein, EGCG, which enhanced the activity of fluoropyrimidines, platinum agents, paclitaxel, doxorubicin, or related antitumor regimens in selected gastric cancer models. However, the evidence base remains heterogeneous and is constrained by variable extract standardization, incomplete dose reporting, poor bioavailability, insufficient pharmacokinetic/pharmacodynamic integration, and underuse of clinically relevant model systems. Overall, medicinal plant bioactives remain promising adjunct candidates in gastric cancer. Still, meaningful translation will require chemically defined interventions, rigorous synergy analysis, interaction-aware study design, and validation in advanced preclinical and clinical settings.

Biomedicines doi: 10.3390/biomedicines14040946

Authors: Yijun Yao Weili Jiang Xinyue Yang Jianyong Wang Ruisi Tang Yuan Feng Yiming Li Mao Chen

Background/Objectives: Transcatheter aortic valve implantation (TAVI) in patients with bicuspid aortic valve (BAV) remains associated with higher rates of residual paravalvular leak (PVL), which confers a two-fold increase in mortality. Despite procedural optimization including balloon post-dilatation, a subset of patients exhibit residual ≥moderate PVL. Pre-procedural identification of these patients could guide procedural planning. Methods: We retrospectively analyzed 402 BAV patients who underwent TAVI with self-expanding valves and balloon post-dilatation between January 2016 and June 2024. A multi-modal deep learning model (Model B) was developed, integrating a 3D ResNet encoder for computed tomography (CT) imaging features with a multilayer perceptron (MLP) for clinical variables, fused via a cross-attention mechanism. Its performance was compared against a conventional model (Model A) combining clinical variables with manually derived CT measurements. Both models were evaluated on identical test folds using 5-fold stratified cross-validation. Results: Of 402 patients, 36 (9.0%) had residual ≥moderate PVL, associated with significantly larger aortic root dimensions at all anatomical levels and greater aortic valve calcification volume (median 887.6 vs. 559.2 mm3; p = 0.004). Model A achieved a mean AUC of 0.694 (95% CI: 0.596–0.792). Model B achieved a mean AUC of 0.822 (95% CI: 0.680–0.964), with a specificity of 0.971, accuracy of 0.881, and PPV of 0.860, while sensitivity was 0.429, reflecting the limited number of outcome events in this cohort. Conclusions: A multi-modal deep learning model integrating expert-segmented CT imaging with clinical variables demonstrated significantly improved discrimination over the conventional approach in this internal cohort for predicting residual PVL in BAV-TAVI, supporting the integration of segmentation-guided deep learning into pre-procedural TAVI planning. However, given the modest number of outcome events, external validation is required to confirm the generalizability of these findings.

Biomedicines doi: 10.3390/biomedicines14040945

Authors: Daniël P. C. van der Spek Renée H. Hoffenkamp Frank J. P. M. Huygen Krishna D. Bharwani Niels Eijkelkamp Maaike Dirckx

Objectives: Complex regional pain syndrome (CRPS) is a heterogeneous pain disorder with incompletely understood immunoinflammatory features. This study investigated whether autonomic receptor autoantibodies differentiate CRPS from other chronic pain conditions and healthy controls. Methods: We conducted a cross-sectional analysis of serum samples from patients referred with suspected CRPS. Patients were subsequently classified as having either CRPS or another chronic pain condition, based on the Budapest criteria. Healthy controls were included for comparison. Serum levels of autoantibodies targeting the muscarinic M2 receptor (M2R), β1-adrenergic receptor (β1AR), and the β2-adrenergic receptor (β2AR) were assessed using enzyme-linked immunosorbent assay. All analyses were performed blinded to group assignment. Results: Seventy participants were analyzed (CRPS = 22, other chronic pain = 25, healthy controls = 23). M2R autoantibody levels were higher in both CRPS and other chronic pain compared with healthy controls (mean difference [MD] = 0.37, 95%CI 0.22–0.51; and MD = 0.31 95%CI 0.19–0.44, respectively). β2AR levels were higher in other chronic pain compared with healthy controls (MD = 0.29, 95%CI 0.04–0.54), whereas no significant difference was observed in CRPS (MD = 0.21 95%CI −0.01–0.42). No meaningful differences were observed between CRPS and other chronic pain for any receptor. β1AR levels did not differ between groups. Seropositivity for any autoantibody was 55% in CRPS, 44% in other chronic pain, and 22% in healthy controls. Conclusions: Elevated autonomic receptor autoantibody levels were observed across chronic pain conditions but were not specific for CRPS.

Biomedicines doi: 10.3390/biomedicines14040944

Authors: Zhiyue Zou Pan Gao Qian Yuan Zhiwen Wang Pengli Luo Chun Zhang

Diabetic nephropathy and diabetic atherosclerosis often develop together and share similar metabolic disturbances. Lipid abnormalities are common in diabetes, yet their roles in kidney and vascular injury are not fully understood. In diabetic kidney disease, altered lipid uptake, reduced fatty acid oxidation, and accumulation of harmful lipid species contribute to cellular stress, mitochondrial injury, inflammation, and fibrosis. In parallel, disordered lipid handling in the vasculature promotes endothelial dysfunction and atherosclerotic plaque development. However, not all lipid accumulation appears to be detrimental, and some findings suggest adaptive or context-dependent effects, leading to inconsistent results across studies. In this review, we summarize current evidence on lipid metabolism in diabetic nephropathy and atherosclerosis, compare shared and distinct features, and discuss ongoing controversies. We also briefly address the therapeutic relevance of targeting lipid pathways and highlight areas that require further investigation. Compared with prior reviews that mainly discussed fatty kidney as an emerging concept in chronic kidney disease research, this review specifically focuses on diabetic kidney disease and integrates kidney-specific lipid trafficking, kidney–vessel crosstalk, conflicting evidence, and mechanism-based therapeutic implications.

Biomedicines doi: 10.3390/biomedicines14040943

Authors: Zijian Song Likai Ren Hong Wang Yanqing Wang Xinxing Du Wei Zhou Qi Zhang Jiyuan Yu Zaixu Zhao Linxiong Ye Kaidi Jin Ying Liu Wei Xue

Background: Bone metastasis is a major determinant of morbidity and therapeutic failure in advanced prostate cancer (PCa); however, the transcriptional programs and tumor microenvironmental alterations driving metastatic progression remain incompletely understood. This study aimed to systematically characterize transcriptomic differences between non-metastatic and bone-metastatic PCa and to identify key microenvironmental signaling pathways involved in tumor survival and chemoresistance. Methods: Bulk RNA sequencing was performed on 49 non-metastatic and 28 bone-metastatic PCa specimens. Differential expression analysis was integrated with weighted gene co-expression network analysis (WGCNA), gene set enrichment analysis, and immune/stromal deconvolution. Key findings were validated using in vitro functional assays, including Transwell co-culture models, small interfering RNA (siRNA)-mediated gene silencing, cell viability, apoptosis, and docetaxel resistance analyses. Results: Transcriptomic profiling identified 574 differentially expressed genes. Bone-metastatic tumors were enriched in ribosome-related and translational pathways, whereas non-metastatic tumors displayed immune-associated signatures, including natural killer (NK) cell-mediated cytotoxicity and cytokine signaling. WGCNA revealed immune-related gene modules preferentially enriched in non-metastatic disease. Immune deconvolution demonstrated significantly higher infiltration of NK cells and endothelial cells in non-metastatic tumors. Chemokine-receptor analysis highlighted upregulation of the CXCL10-CXCR3 axis in non-metastatic PCa. In vitro, PCa cells expressed CXCR3, while endothelial cells markedly increased CXCL10 expression upon co-culture. Functional assays showed that endothelial-derived CXCL10 promoted PCa cell survival, suppressed apoptosis, and conferred resistance to docetaxel via CXCR3-dependent signaling; these effects were reversed by CXCL10 or CXCR3 knockdown. Conclusions: These findings uncover a context-dependent endothelial-immune chemokine network distinguishing non-metastatic from bone-metastatic PCa and identify the CXCL10-CXCR3 axis as a critical mediator of tumor survival and chemoresistance, suggesting a potential therapeutic vulnerability in advanced prostate cancer.

Biomedicines doi: 10.3390/biomedicines14040942

Authors: Paweł Głuszak Julia Woźna Andrzej Bałoniak Jakub Pazdrowski Jan Stępka Joanna Kaźmierska Aleksandra Dańczak-Pazdrowska

Background: Acute radiation-induced skin injury (aRISI) is one of the most frequent adverse effects of radiotherapy (RT) in patients with head and neck cancer (HNC) and may compromise treatment delivery and quality of life. Topical glucocorticosteroids (GCS) are commonly used in clinical practice for aRISI management; however, evidence supporting their proactive use remains inconsistent. This systematic review and meta-analysis aimed to evaluate the efficacy and safety of proactive topical GCS therapy during RT for HNC. Methods: A systematic search of PubMed, Embase, and the Cochrane Library was conducted from database inception to July 2025 in accordance with PRISMA 2020 guidelines. Randomized controlled trials comparing topical GCS with placebo or standard skin care in adult patients undergoing curative RT or RChT for HNC were included. The primary outcomes were incidence of clinically significant aRISI (grade ≥ 2) and severe aRISI (grade ≥ 3), assessed using validated grading systems (RTOG or CTCAE). Random-effects meta-analyses were performed to calculate pooled risk ratios (RRs) with 95% confidence intervals (CIs). Risk of bias was assessed using the Cochrane RoB 2 tool. Results: Three randomized controlled trials comprising 459 patients were included. Proactive topical GCS did not significantly reduce the pooled incidence of grade ≥ 2 aRISI compared with placebo or standard skin care (RR 0.87, 95% CI 0.60–1.27). Similarly, no statistically significant reduction in grade ≥ 3 aRISI was observed in pooled analysis (RR 0.81, 95% CI 0.22–3.06). Qualitative synthesis of secondary outcomes reported in individual trials suggested potential benefits of topical GCS, including delayed onset or slower progression of aRISI, and, in one large double-blind study, a reduced risk of severe reactions. No increase in treatment-related adverse events was observed in any included trial. Conclusions: Proactive topical GCS do not significantly reduce the overall incidence of aRISI in pooled analysis. Individual studies showed trend towards delayed onset, slower progression, and reduced severe aRISI without compromising safety. These findings support the judicious use of topical GCS as part of proactive supportive care in HNC RT, while highlighting the need for larger, standardized trials to define optimal regimens and patient selection.

Biomedicines doi: 10.3390/biomedicines14040941

Authors: Magdalena Potapowicz-Krysztofiak Martyna Dąbrowska Małgorzata Maciorowska Zbigniew Orski Paweł Krzesiński Marek Kiliszek Beata Uziębło-Życzkowska

Background: Left bundle branch block (LBBB) and QRS prolongation are markers of electrical dyssynchrony in heart failure, but they do not fully reflect its mechanical consequences. Myocardial work (MW)-derived indices may provide a more comprehensive assessment of left ventricular (LV) mechanical dyssynchrony. We evaluated associations between LV MW parameters, QRS duration, and LBBB in patients with heart failure with reduced ejection fraction (HFrEF) referred for ICD/CRT implantation. Methods: In this single-centre observational cross-sectional study, 96 consecutive patients referred for ICD or CRT implantation were screened. All patients underwent standardized baseline comprehensive echocardiography followed by advanced MW analysis. Myocardial work index (MWI) dispersion was assessed using two complementary methods. MWI dispersion (SD) was calculated as the standard deviation of segmental MWI values across all LV segments, and MWI dispersion (IQR) was defined as the interquartile range (IQR) of segmental MWI values. We evaluated the associations between QRS duration and MW-derived dyssynchrony parameters (individual and composite), as well as their discriminative performance for LBBB. Seven patients were excluded from further analysis due to inadequate echocardiography image quality. Results: The final study group comprised 89 patients with HFrEF (median age 65.5 years), of whom 67.4% were assigned to CRT. LBBB was present in 41.6%, and the median QRS duration was 142 ms (112–162). All analyzed LV MW indices were significantly associated with QRS duration (all q < 0.01). The strongest correlations were observed for MWI dispersion (IQR) (r = 0.58), peak strain dispersion (PSD) (r = 0.54), lateral–septal work asymmetry (r = 0.53), and MWI dispersion (SD) (r = 0.52) (all q < 0.0001). All MW indices differed significantly between patients with and without LBBB (all q ≤ 0.0001). MWI dispersion (IQR) showed the best single-marker discrimination of LBBB (AUC = 0.852). Composite indices achieved AUC = 0.84 but did not significantly improve discrimination versus MWI dispersion (IQR) alone. Conclusions: Myocardial work-derived indices of left ventricular dyssynchrony are strongly associated with QRS duration and the presence of LBBB in patients with HFrEF. Among them, MWI dispersion (IQR) was shown to be the best-performing MW marker for identifying LBBB. These findings suggest that MW dispersion may serve as a robust echocardiographic marker of mechanical dyssynchrony and warrants further investigation as a potential tool for predicting CRT response.

Biomedicines doi: 10.3390/biomedicines14040940

Authors: Nur Akman Ahmet Ufuk Kömüroğlu Salih Çibuk Fikret Altındağ Osman Yılmaz Ahmet Ateşşahin

Background/Objectives: Autism spectrum disorder (ASD) is a neurodevelopmental condition increasingly associated with dysregulated neuroimmune signaling and altered neurotrophic homeostasis. Tumor necrosis factor-alpha (TNF-α) has been implicated in ASD pathophysiology; however, the downstream effects of TNF-α blockade on cytokine–neurotrophin interactions during neurodevelopment remain insufficiently characterized. In this study, we evaluated the effects of infliximab (IFX), a monoclonal anti-TNF-α antibody, on behavioral performance, neuroinflammatory cytokine profiles, glial activation, and brain-derived neurotrophic factor (BDNF) signaling in a propionic acid (PPA)-induced experimental ASD rat model. Methods: Experimental ASD was induced by propionic acid administration in rats. Animals were divided into control and treatment groups. Behavioral performance was assessed using the Morris Water Maze, direct social interaction, and three-chamber sociability tests. Levels of TNF-α, interleukin-1 beta (IL-1β), interleukin-6 (IL-6), and BDNF were measured in serum, hippocampal, and cerebellar tissues. Microglial and astrocytic activation were evaluated using CD11 and GFAP immunohistochemistry. Results: PPA administration resulted in pronounced impairments in learning, memory, and social behaviors, accompanied by elevated proinflammatory cytokine levels, increased BDNF expression, and marked glial activation in the hippocampus and cerebellum. Although IFX treatment significantly reduced TNF-α levels in central tissues, it did not improve behavioral deficits and was associated with persistently elevated IL-1β and IL-6 levels, sustained glial reactivity, and further alterations in BDNF levels. Conclusions: These findings suggest that TNF-α suppression alone does not normalize the disrupted cytokine–neurotrophin axis and may differentially modulate BDNF-related neuroplastic signaling during development. In conclusion, this study indicates that non-selective TNF-α blockade during neurodevelopment fails to confer behavioral benefit in experimental ASD and highlights the importance of considering cytokine–BDNF pathway interactions when designing immunomodulatory strategies for neurodevelopmental disorders.

Biomedicines doi: 10.3390/biomedicines14040939

Authors: Yu Gao Yuanyuan Cao Junyao Liu Fang Tong Xianlin Liu Jiahui Wang Peixuan Liu Yanting Xu Lu Feng Pengxin Zhang Jingchun Gao Jiwei Liu

Background/Objectives: The clinical challenges of PARP inhibitors in ovarian cancer include the lack of effective maintenance regimens for homologous recombination proficiency (HRP) patients and the emergence of acquired resistance in initially responsive homologous recombination deficiency (HRD) patients. This study aims to explore the synergistic effect and molecular mechanism of the bispecific tyrosine phosphorylation-regulated kinase 1B (DYRK1B) inhibitor AZ191 combined with the PARP inhibitor Niraparib on high-grade serous ovarian cancer (HGSOC). Methods: This study first explored the expression and prognostic significance of DYRK1B in ovarian cancer through bioinformatics analysis. Subsequently, the therapeutic effect of the DYRK1B inhibitor AZ191 combined with Niraparib on HGSOC cells and organoids was evaluated by MTT examination. Flow cytometry and Western blot were used to investigate the synergistic mechanism between the two agents. Results: Bioinformatics analysis shows that the high expression of DYRK1B in serous ovarian cancer is associated with poor prognosis of the patients. The experiments in vitro have shown that the DYRK1B inhibitor AZ191 can enhance the therapeutic effect of Niraparib on HGSOC cells and organoids, whether HRD-positive or not. Mechanistic studies have shown that the combination of AZ191 and Niraparib can synergistically increase the accumulation of DNA damage, thereby intensifying the apoptosis of HGSOC cells. In addition, the combination therapy induces ferroptosis by inhibiting the Nrf2/SLC7A11/GPX4 axis, thereby exerting cytotoxic effects. Conclusions: Our results uncover a novel mechanism by which inhibiting DYRK1B enhances the anti-HGSOC efficacy of Niraparib and may offer a promising treatment strategy to improve the maintenance therapy in both HRD and HRP ovarian cancer patients.

Biomedicines doi: 10.3390/biomedicines14040938

Authors: Adrian-Florentin Dragomir Smaranda Stoleru Aurelian Zugravu Elena Poenaru Maria Carina Dumitrescu Aurelia Cristiana Barbu Silvia Fratea Clara Maria Stoleru Oana Andreia Coman Ion Fulga

Background/Objectives: Cholinergic dysfunction plays a central role in memory impairment, yet trihexyphenidyl (THP)-based paradigms remain less explored than scopolamine-based models. This study aimed to characterize a THP-induced cholinergic challenge in a two-trial Y-maze with a 24 h interval and to compare the effects of donepezil and tacrine on recall-phase exploratory allocation. Methods: Male Wistar rats (n = 9/group) were studied in a validation phase including saline, THP 5 mg/kg, and THP 10 mg/kg groups, followed by an intervention phase including control, THP 10 mg/kg, donepezil 1 and 3 mg/kg + THP, and tacrine 3 and 5 mg/kg + THP groups. All treatments were administered intraperitoneally (i.p.). Acquisition- and recall-phase behavior was analyzed. Recall outcomes included arm times, arm entries, the novel-to-familiar arm time ratio (U/K time ratio), the novel-to-familiar arm entry ratio (U/K entry ratio), discrimination indices and time-per-entry indices. Data were analyzed by one-way ANOVA; Tukey’s post hoc test was used in the validation experiment, whereas Dunnett’s test was used in the intervention experiment for comparisons against THP 10. Results: THP at 10 mg/kg produced a robust recall-phase phenotype, with increased familiar-arm exploration, reduced novel-arm exploration and lower normalized indices. Under THP challenge, donepezil was associated with clearer effects at 3 mg/kg, whereas tacrine displayed a broader dose-dependent profile, with the strongest shift in recall-phase exploratory allocation toward the novel arm observed at 5 mg/kg. Conclusions: THP 10 mg/kg produced a robust recall-phase exploratory phenotype in a 24 h two-trial Y-maze paradigm. Under THP challenge, donepezil and tacrine were associated with shifts in recall-phase exploratory allocation. These findings support the potential utility of THP-based paradigms for studying cholinergic disruption in Y-maze settings, while direct comparison with scopolamine-based models remains to be established.

Biomedicines doi: 10.3390/biomedicines14040937

Authors: Maja Štrajtenberger Marinko Artuković Iva Bešlić Morana Belovic Ivica Lokner Liborija Lugović-Mihić

Background: The interaction between coagulation pathways, inflammatory markers, and hematological parameters has not been sufficiently clarified in patients with chronic angioedema (AE) and urticaria. This study aimed to validate previously observed associations and to further explore their relationship with clinical disease control. Methods: In this cross-sectional validation study, 102 participants were enrolled and stratified into three groups: isolated AE (n = 33), AE associated with urticaria (AE/Urt; n = 34), and healthy controls (n = 35). Serum levels of coagulation factors (D-dimer, fibrinogen, factor VII), inflammatory markers (C-reactive protein [CRP], erythrocyte sedimentation rate [ESR]), and complete blood count parameters were analyzed. Disease control was assessed using the Angioedema Control Test (AECT). Appropriate non-parametric statistical tests were applied. Results: Only D-dimer values differed significantly between groups and were higher in patients with AE/Urt than in controls. At the same time, D-dimers were significantly more often elevated in both AE groups than in healthy individuals. Additionally, CRP values in both AE groups were significantly more often elevated than in controls, with significantly higher values in both AE groups (in both groups 85%) than in controls (57%). Coagulation markers and CRP demonstrated a positive correlation with age (r = 0.268–0.392; p ≤ 0.007), with fibrinogen of all coagulation markers showing the strongest age dependency (r = 0.334; p = 0.001). Gender-related differences in coagulation parameters were not statistically significant, although elevated fibrinogen levels were more common in male participants (p = 0.030). Disease control did not correlate linearly with any inflammatory markers, coagulation factor, age or gender. Conclusions: The findings support a contributory role of coagulation pathway activation and systemic inflammation in the pathophysiology of chronic angioedema and urticaria. These mechanisms may influence clinical disease expression and could represent potential targets for improved diagnostic stratification and therapeutic approaches. However, the interpretation of the present results should be approached with caution in light of several important study limitations, including demographic heterogeneity between the study groups and the relatively limited sample size.

Biomedicines doi: 10.3390/biomedicines14040935

Authors: Ragaey Ahmad Eid Michael Nady Naguib Amr Ahmed Abd El Bary Mohamed Medhat Mohamed Zaki Marwa O. Elgendy Anwar M. Alnakhli Mohammed Gamal Mohamed Mohamed Tawfik

Background/Objectives: Non-variceal upper gastrointestinal bleeding (NVUGIB) remains a major clinical emergency, particularly among patients receiving antiplatelet or anticoagulant therapy, whose use has increased substantially in recent years. This study aimed to evaluate the clinical characteristics, endoscopic findings, risk stratification, and outcomes of NVUGIB in patients receiving antithrombotic therapy, and to compare the predictive performance of commonly used prognostic scores. Methods: This prospective cohort study included 89 patients receiving antithrombotic therapy who presented with NVUGIB at Beni-Suef University Hospitals between March 2023 and March 2025. Clinical presentation, laboratory findings, and endoscopic characteristics were recorded. Risk stratification was assessed using Glasgow–Blatchford (GBS), Rockall, Baylor, AIMS65, ABC, and PNED scores. The optimal cut-off values for prediction of rebleeding and mortality were determined using receiver operating characteristic (ROC) analysis and the Youden index. Area under the curve (AUC) values were reported with 95% confidence intervals. Results: Endoscopy revealed that peptic ulcers were the most common lesion (41/89, 46%), followed by erosive disease (27/89, 30%), with the stomach being the most frequently involved site (76.5%). Rebleeding occurred in 16 patients (18.0%), while mortality was observed in 2 patients (2.2%). The Glasgow–Blatchford score demonstrated the most consistent performance for predicting rebleeding, with an optimal cutoff value of 5.5 (derived using the Youden index), yielding 92.9% sensitivity and 78.8% specificity. For mortality prediction, AIMS65, ABC, and PNED scores showed very high AUC values, although these findings should be interpreted cautiously due to the small number of mortality events (n = 2). No statistically significant difference in rebleeding or mortality was observed between single and dual antithrombotic therapy, although patients receiving dual therapy required longer hospitalization and more transfusion units. Conclusions: In patients with antithrombotic-related GI bleeding, ulcers and erosions predominate, with minimal differences between single and dual therapy outcomes. Concomitant NSAID use trends toward higher mortality. Glasgow–Blatchford score offers optimal performance for both rebleeding and mortality prediction, with a cutoff of 5.5 providing excellent sensitivity (92.9%) and specificity (78.8%) for rebleeding risk assessment.

Biomedicines doi: 10.3390/biomedicines14040936

Authors: Seyed Mojtaba YazdanParast Navid Manoochehri Mohammad Abdolahad

Background: Electrochemotherapy enhances the intracellular delivery of anticancer agents through electroporation but is traditionally limited to cytotoxic drugs associated with significant side effects. Vitamin C (ascorbic acid) exhibits selective anticancer activity when accumulated at high intracellular concentrations; however, its therapeutic application is restricted by poor membrane permeability and rapid systemic clearance. Methods: In this study, we investigated whether reversible electroporation, applied using a custom-designed variable plate electrode system designed to deliver a uniform electric field, could potentiate the antitumor efficacy of low-dose vitamin C. Numerical simulations were performed to optimize electrode spacing and stimulation voltage, suggesting homogeneous electric field coverage throughout the tumor volume. The proposed approach was evaluated in vitro using MDA-MB-231 and 4T1 breast cancer cell lines and in vivo in a 4T1 murine breast cancer model. Results: Low-dose vitamin C alone produced minimal cytotoxic effects, whereas its combination with electroporation significantly reduced cell viability and increased apoptotic and necrotic cell death in vitro. In vivo, vitamin C–assisted electrochemotherapy resulted in pronounced tumor growth suppression, with tumor volumes reduced to approximately 0.34-fold of baseline by day 15, accompanied by decreased proliferation and marked tissue disruption. Conclusions: These findings demonstrate that uniform-field reversible electroporation markedly enhances the intracellular delivery and antitumor activity of low-dose vitamin C, supporting this technology-driven strategy as a promising, low-toxicity alternative to conventional chemotherapeutic agents in electrochemotherapy for solid tumors.

Biomedicines doi: 10.3390/biomedicines14040934

Authors: Sanyukta Bihari Dia Yang Devarshi Mukherji Aya Haggiagi

Diffuse midline gliomas (DMGs) are rare but highly aggressive central nervous system (CNS) tumors that can present in both pediatric and adult populations. These tumors were redefined in the 2016 WHO classification of CNS tumors based on integrated histopathological and molecular features, and were initially designated as “DMG, H3 K27M-mutant”. In the 2021 WHO update, DMGs were incorporated into the newly defined category of primarily pediatric-type diffuse high-grade gliomas, and nomenclature was changed to “DMG, H3 K27-altered” to encompass additional molecular drivers beyond the canonical H3 K27M mutation. Clinically, DMGs arise as expansile, infiltrating tumors within midline structures and may present as non-enhancing or enhancing lesions on imaging. Diagnosis is based on neuroimaging and molecular confirmation by immunohistochemistry or sequencing when tissue is available. DMGs are categorized as WHO grade 4 malignant tumors due to their aggressive biology leading to rapid and infiltrative growth. Owing to their deep and midline location, surgical resection is typically not feasible. Radiation therapy is the backbone of treatment, but there is no standard regimen of chemotherapy that has demonstrated durable efficacy. Recent progress in therapeutic approaches has led to a major breakthrough on 6 August 2025 when the U.S. Food and Drug Administration granted the accelerated approval of dordaviprone (ONC201), marking it as the first systemic therapy for progressive DMG harboring H3 K27M mutation. Other novel approaches, including chimeric antigen receptor (CAR) T-cell directed therapies and convection-enhanced delivery, are actively under investigation. We aim to comprehensively review DMGs, including the recent insights into their biology, the evolving therapeutic landscape, and the opportunities to fuel this new momentum against one of the most formidable gliomas.

Biomedicines doi: 10.3390/biomedicines14040933

Authors: Amanda Balboa Ramilo Kevin Mani Anders Wanhainen Henrik Lodén Anna Nilsson Per E. Andrén Malou Friederich-Persson Dick Wågsäter

Background and aims: Abdominal aortic aneurysm (AAA) is a vascular disease characterized by the progressive dilation of the aorta, culminating in rupture. At present, there are no pharmacological treatments to prevent AAA development or reduce rupture rate. A recent study showed that patients prescribed Glucagon-like peptide-1 receptor agonists (GLP-1RAs) have significantly lower risks of mortality, AAA repair, and acute abdominal aortic syndrome. Semaglutide is a GLP-1RA with increased agonist capacity and longer half-life, compared to earlier generations of GLP-1RAs. In this study, we aimed to investigate the role and mechanisms of semaglutide in the prevention of AAA development and rupture in a murine model. Methods: AAA was induced in apolipoprotein-E-deficient mice, by continuous subcutaneous infusion of angiotensin II. Treatment with semaglutide (12 µg/kg) began seven days after disease induction (rescue trial) or simultaneously with disease induction (prophylactic trial). At experimental endpoint, aortic diameter was measured by high-frequency ultrasound and the aortic tissue was collected for histological analysis. Results: Prophylactic treatment with semaglutide drastically reduced mortality by dissection and rupture during the first seven days of disease development, but did not affect AAA formation at 28 days. Histological evaluation of the aorta at day seven showed a normal vessel wall thickness with a trend for a higher content of collagen in the aortic wall in mice treated with semaglutide, compared to controls. Conclusions: Semaglutide prevents aortic rupture and dissection in the early phases of AAA development in the angiotensin II mouse model, likely by promoting the maintenance of an adequate proportion of collagen in the vessel wall.

Biomedicines doi: 10.3390/biomedicines14040932

Authors: Basmah Eldakhakhny Sumia Enani Hanan Jambi Ghada Ajabnoor Jawaher Al-Ahmadi Rajaa Al-Raddadi Lubna Alsheikh Wesam H. Abdulaal Hoda Gad Anwar Borai Suhad Bahijri Jaakko Tuomilehto

In the original publication [...]

Biomedicines doi: 10.3390/biomedicines14040931

Authors: Toar Jean Maurice Lalisang Vania Myralda Giamour Marbun Linda Erlina Nathaniel Jason Zacharia Kezia Nathania Limbong Allo Fadilah Fadilah Aisyah Fitriannisa Prawiningrum

Background/Objectives: Population-specific genomic data are essential for understanding hepatocellular carcinoma (HCC) biology, particularly in underrepresented regions. This study aimed to perform exploratory single-nucleotide polymorphism (SNP)-array-based profiling of HCC tumor samples from Indonesian patients and to prioritize candidate functional variants using a systematic in silico framework. Methods: This retrospective cross-sectional study included 15 resected HCC cases with available formalin-fixed paraffin-embedded (FFPE) tumor tissue. Genome-wide SNP genotyping was performed using the Illumina Asian Screening Array. Following quality control and filtering, variants were annotated using the Ensembl Variant Effect Predictor. A case-only functional prioritization approach incorporating multiple in silico prediction tools was applied, followed by gene-level burden aggregation. Results: After multistep filtering, 11 samples and 104 prioritized variants were retained for analysis. Variants consisted predominantly of splice-region, missense, and regulatory changes. Gene-level burden analysis identified Nuclear Autoantigenic Sperm Protein (NASP, rs775916096) as the highest-ranked candidate gene, while G protein-coupled receptor 78 (GPR78, rs558447540) emerged as a secondary candidate with predicted functional annotations but currently limited biological evidence in HCC. Given the tumor-only design without matched normal tissue, the prioritized variants cannot be distinguished from rare germline variants. Conclusions: This exploratory SNP-array study provides a hypothesis-generating framework for functional variant prioritization in Indonesian HCC. NASP and GPR78 represent preliminary candidates that require validation in larger cohorts with matched normal tissue and sequencing-based confirmation.

Biomedicines doi: 10.3390/biomedicines14040930

Authors: Julian Kreutz Klevis Mihali Vivien Sievertsen Lukas Harbaum Georgios Chatzis Styliani Syntila Bernhard Schieffer Birgit Markus

Background: Post-cardiac arrest syndrome (PCAS) following out-of-hospital cardiac arrest (OHCA) is driven by global ischemia–reperfusion injury, endothelial dysfunction, and a dysregulated inflammatory response. This cascade frequently culminates in profound vasoplegia and multiorgan failure, even when guideline-directed post-resuscitation management is applied. Hemoadsorption using the CytoSorb device may attenuate hyperinflammation and vasoplegia by removing circulating inflammatory and injury-related mediators. Methods: This single-centre, retrospective cohort study compared adults with PCAS following OHCA who received hemoadsorption with propensity score-matched controls (1:1 matching; n = 50 per group). For patients treated with hemoadsorption, data were analyzed within predefined intervals covering the 24 h preceding therapy initiation (T1) and the 24 h following the completion of the hemoadsorption treatment period (T2). Controls were evaluated at time points aligned to those of their matched hemoadsorption counterparts. Hemodynamic, metabolic, respiratory, and organ injury markers were assessed. Results: Formal between-group comparisons of temporal change between T1 and T2 showed no statistically significant differences between hemoadsorption-treated patients and matched controls across key parameters, including VIS (Δ −18.7 vs. −7.7; p = 0.183) and lactate (Δ −1.8 vs. −1.25 mmol/L; p = 0.780), as well as markers of organ injury, pH, and oxygenation. In exploratory ANCOVA models, only base excess was associated with treatment group (p = 0.035). Survival to hospital discharge was comparable (48% vs. 40%; p = 0.423), with similar neurological outcomes. Within the hemoadsorption group, pre–post comparisons around hemoadsorption initiation (T1–T2) demonstrated marked improvements, including reduced vasoactive support (VIS 70.0 to 12.1; p = 0.039), substantial lactate clearance (4.1 to 1.1 mmol/L; p < 0.001), and declines in organ injury markers (AST, ALT, LDH, myoglobin), alongside more pronounced platelet reduction compared with controls (129 to 57 × 103/µL vs. 189 to 123 × 103/µL). However, adjusted analyses indicated that these changes were primarily driven by baseline shock severity rather than a treatment-specific effect. Conclusions: In this propensity score-matched cohort of PCAS patients after OHCA, hemoadsorption was associated with within-group physiological changes but showed no detectable advantage over matched controls, with similar survival. These findings are hypothesis-generating and warrant prospective studies with standardized timing and phenotype-guided patient selection.

Biomedicines doi: 10.3390/biomedicines14040929

Authors: Jianan Shen Aihua Yang Xintao He Tianyi Dai Zexuan Hui Youxiang Ding Li Zhao Jun Chen

Background: The immunologically cold nature and immunosuppressive tumor microenvironment (TME) of intrahepatic cholangiocarcinoma (ICC) contribute to its poor prognosis. This study aims to identify novel biomarkers related to prognosis and TME in ICC. Methods: We first identified the high expression of m6A reader insulin-like growth factor 2 mRNA binding protein 2 (IGF2BP2) in ICC through bioinformatics screening. Subsequently, a retrospective study was conducted on 224 ICC patients who had undergone radical resection. The expression levels of IGF2BP2 and programmed death ligand 1 (PD-L1) were detected in a tissue microarray (TMA) using immunohistochemistry (IHC). The co-localization of IGF2BP2, PD-L1, programmed cell death protein 1 (PD-1), and CD8+T cells was evaluated by multiple immunofluorescence techniques. Results: IHC confirmed a significant upregulation of IGF2BP2 in tumor tissues compared with normal bile duct epithelia (p < 0.05). IGF2BP2 expression was positively correlated with PD-L1 expression (TPS R = 0.215, p = 0.016; CPS R = 0.295, p = 0.008). High IGF2BP2 expression was associated with increased PD-L1/PD-1 positivity and reduced CD8+T cell infiltration. Kaplan–Meier analysis revealed significantly worse 3-year overall survival (OS: 20.56% vs. 29.91%, p = 0.0291) and recurrence-free survival (RFS: 9.72% vs. 18.56%, p = 0.0372) in the IGF2BP2-high group. Multivariate analysis identified IGF2BP2 as an independent risk factor for both OS (HR = 1.683, p = 0.044) and RFS (HR = 1.946, p = 0.042). Conclusions: IGF2BP2, as a potential biomarker and independent prognostic factor for ICC, is associated with increased PD-L1 expression.

Biomedicines doi: 10.3390/biomedicines14040928

Authors: Martina Perše

The dextran sodium sulfate (DSS) colitis model is the most widely used experimental model of inflammatory bowel disease (IBD) due to its simplicity and versatility, with over 7000 PubMed entries in the last decade and an exponential rise in recent years. Since its initial description in 1985, DSS colitis has been extensively evaluated across species, most notably in mice and rats, and has yielded substantial insights into IBD pathogenesis. However, the model’s multifactorial nature poses a dual challenge: it offers an opportunity but complicates study design, interpretation, and translational relevance. This complexity is worsened by inconsistent reporting, which hampers reproducibility and comparability across studies. The broad use of the DSS-induced colitis model yields numerous insights about the model, which help better understand its complexity, characteristics and limitations. Although DSS colitis is induced locally, inflammation in the colon and gut barrier destruction may also affect other organs (such as the liver and brain) and their metabolism and molecular responses, which, in turn, may interfere with colitis-underlying mechanisms and drug response, and may influence the interpretation of results. These intrinsic (intra-experimental) characteristics of the DSS model are summarised in the paper (colitis, gut–brain axis, gut–liver axis). In addition, the DSS model is heavily influenced by numerous extrinsic (inter-experimental) factors (environmental, microbiological, genetic), which may further complicate the colitis model, the study outcomes, and data interpretation, and these are also discussed in the paper. As science advances and new data accumulate, understanding the intricate interplay among internal mechanisms, external factors, and technical variables becomes increasingly essential for the accurate interpretation of DSS outcomes. This review synthesises the complexity and interdependence of factors shaping the DSS model, emphasising the need for meticulous reporting and consideration of methodological nuances to enhance reproducibility, interpretation, and translational value in DSS colitis research. In addition, the review provides practical guidance through a “traps and tricks” subsection and checklist table designed to provide a framework and practical recommendations to better understand, apply, and interpret DSS model results in the context of broader systemic and methodological considerations.

Biomedicines doi: 10.3390/biomedicines14040927

Authors: Xiaoguang Liu Sean Baxely Michaeline L. Hebron Charbel Moussa

Introduction: Pathogenic mutations in leucine-rich repeat protein kinase-2 (LRRK2), particularly G2019S, constitute the most common cause of autosomal dominant PD. Methods: Mouse models encoding human mutant alpha-synuclein (SNCA A53T) and LRRK2 G2019S were treated with a brain-penetrant kinase inhibitor (BK40196). Behavior, nigrostriatal and mesolimbic dopamine (DA) pathways were examined. Results: Mice harboring LRRK2 G2019S do not show age-dependent motor symptoms, but mice encoding SNCA A53T display motor deficits, while both strains exhibit anxiety-like behavior and BK40196 improves motor and behavioral defects. BK40196, a multi-kinase inhibitor of Abelson (Abl), Discoidin domain receptor (DDR)-1, c-KIT and FYN, alters microglial morphology and alpha-synuclein levels in SNCA A53T mice and improves DA neurotransmission, primarily via the nigrostriatal system. BK40196 inhibits brain LRRK2 G2019S (IC50 of 89nM) and does not affect phosphorylated or total peripheral LRRK2 levels (lungs, kidneys, liver, etc.). LRRK2 G2019S mice treated with BK40196 exhibit distinct increases in DA in mesolimbic neurons such as the nucleus accumbens (NAcc), suggesting differential mechanisms of DA neurotransmission in mutant alpha-synuclein and LRRK2 models of PD. Conclusions: LRRK2 G2019S may primarily involve mesolimbic pathways leading to nonmotor symptoms independent of the motor and behavioral manifestations associated with alpha-synuclein via the nigrostriatal system. BK40196 may provide a comprehensive and synergistic therapeutic approach that addresses multiple mechanisms to reduce the pathologies related to LRRK2 G2019S and/or SNCA in PD. The multiple pathologies of PD necessitate a holistic approach that simultaneously targets inflammation and autophagy and LRRK2 inhibition.

Biomedicines doi: 10.3390/biomedicines14040926

Authors: Montserrat Fernández-Guarino María Luisa Hernández Bule Stefano Bacci

This perspective contains the current understanding of the cellular and molecular mechanisms involved in wound healing (the articles taken into consideration relate to the three-year period 2023–2025). Nevertheless, these biological pathways remain inadequately characterized; this is seen by the modifications leading to pathological conditions, such as keloids, chronic wounds, or hypertrophic scars and diabetic wounds. Focus is also directed to novel therapy suggested for these types of conditions. Understanding these scientific issues is crucial for professionals across many fields who see such presentations often.

Biomedicines doi: 10.3390/biomedicines14040925

Authors: Raffaele Radice Giulia Pollaroli Michela Salvatici Chiara Corrado Francesca Rispoli Stefania Pacchetti Lorenzo Drago

Background: Primary biliary cholangitis (PBC) is a rare autoimmune liver disease characterized by marked clinical and serological heterogeneity. Although diagnosis is mainly based on antimitochondrial antibodies (AMAs) and alkaline phosphatase (ALP), non-classical presentations remain a relevant cause of diagnostic delay. In this context, laboratory medicine plays a pivotal role in both diagnosis and long-term disease management. Methods: This manuscript represents a structured clinical review of laboratory biomarkers relevant to the diagnosis, monitoring, and prognostic stratification of PBC, integrated with a representative atypical case with long-term follow-up to illustrate the practical application of laboratory-driven diagnostic. Results: The analysis confirms the central role of immunological and biochemical markers in treatment monitoring and prognostic assessment, while highlighting their limitations in selected clinical scenarios. The reported case, characterized by persistent AMA negativity and consistently normal ALP levels, illustrates how expanded laboratory testing can support the identification of non-standard disease phenotypes. In this setting, parallel testing for AMA- and PBC-specific autoantibodies was essential to achieve a correct diagnosis. Moreover, alternative biomarkers, including gamma-glutamyl transferase (GGT) and selected immunological markers, provided clinically meaningful information when conventional markers were not informative. Conclusions: By integrating current evidence with a long-term clinical case, this work moves beyond a descriptive overview and proposes a practical, laboratory-driven diagnostic and follow-up framework for PBC. It highlights laboratory opportunities to facilitate timely diagnosis, appropriate prognostic stratification, and disease monitoring, including the assessment of associated comorbidities.

Biomedicines doi: 10.3390/biomedicines14040924

Authors: Tzu-Rong Peng Hung-Hong Lin Ta-Wei Wu Chin-Yu Hsu

Background/Objectives: While standard antiemetic regimens have evolved, breakthrough symptoms and anticipatory nausea persist. Lorazepam has historically been used as an adjunct, yet a comprehensive re-evaluation of its efficacy across historical trials is lacking. This study provides a synthesis of clinical evidence to re-evaluate the adjunctive therapeutic value of lorazepam, potentially addressing persistent gaps in emesis control, such as anticipatory and refractory symptoms. Methods: Following PRISMA guidelines, we analyzed eight randomized controlled trials (n = 864) published between 1989 and 1997. Primary endpoints included complete and improved responses for emesis and nausea. Results: Eight trials (n = 864), published between 1989 and 1997, met the inclusion criteria. Lorazepam-containing regimens significantly increased the complete response rate for overall emesis (OR = 1.55; 95% CI, 1.12–2.14; p = 0.008) and improved the response rate (OR = 1.50; 95% CI, 1.03–2.19; p = 0.04). Subgroup analysis of acute emesis showed consistent benefits (complete response OR = 1.77; 95% CI 1.23–2.55; p = 0.002). Nausea control also favored lorazepam, although the differences were not statistically significant. Sedation was more frequent with lorazepam (RR = 2.67; 95% CI 1.54–4.63), although no serious adverse events were reported. Conclusions: By revisiting decades of clinical evidence, this meta-analysis confirms that lorazepam provides a significant therapeutic advantage in controlling chemotherapy-related vomiting, particularly during the acute phase. While its direct efficacy alongside modern agents like NK1 receptor antagonists remains to be fully elucidated, the anxiolytic and amnestic properties of lorazepam remain a potential adjunct for managing complex CINV profiles. Careful dose titration is necessary to balance efficacy with sedation.

Biomedicines doi: 10.3390/biomedicines14040917

Authors: Aranza Mejía-Muñoz Jessica Cedillo Monter Héctor Iván Saldívar-Cerón Galileo Escobedo Sonia Leon-Cabrera

Sucralose is one of the most widely used non-nutritive sweeteners and has long been considered metabolically inert and safe within established acceptable daily intake levels. However, emerging evidence suggests that chronic exposure to sucralose may alter gut microbial composition, epithelial barrier function, mucosal inflammation, and immune responses. This review examines current experimental and clinical evidence on the effects of sucralose on the gut–immune axis, with particular attention to its potential implications for colitis and colorectal cancer (CRC). Preclinical studies indicate that sucralose may reduce beneficial short-chain fatty acid-producing taxa, alter microbial metabolic pathways, disrupt epithelial barrier-related molecules, and promote inflammatory and immune changes associated with colitis severity and inflammation-driven tumorigenesis. Experimental evidence also suggests that sucralose may impair CD8+ T-cell fitness and reduce responsiveness to immune checkpoint inhibitors through microbiome-dependent mechanisms involving altered arginine and citrulline metabolism. Human studies further indicate that sucralose can modify gut and oral microbiome composition and influence metabolic responses, although these effects appear heterogeneous and context-dependent. Overall, the current literature suggests that sucralose may act as a modifier of microbiome–immune interactions in susceptible settings, but most mechanistic evidence remains preclinical, and human data are still insufficient to establish causality. These findings highlight the need for prospective studies to determine whether sucralose-associated microbial and immune alterations translate into clinically meaningful effects in colitis, CRC, and immunotherapy response.

Biomedicines doi: 10.3390/biomedicines14040923

Authors: Yulia A. Desheva Tatiana V. Gupalova Polina A. Kudar Galina F. Leontieva Igor V. Kudryavtsev Andrey S. Trulioff Danila S. Guzenkov Victoria A. Matyushenko Elena A. Bormotova Daniil D. Sokolovsky Georgy A. Matveev Boris P. Nikolaev Alexander N. Suvorov

Background/Objective: This study is a cross-sectional investigation of long-term immune responses measured at different time intervals after COVID-19 infections, vaccinations, or combined exposure. The focus is on immune reactivity against recombinant spike (S) and nucleocapsid (N) protein antigens. Materials and Methods: Serum antibody levels were assessed up to four to four and a half years after infection or immunization, including virus-specific immunoglobulin G (IgG), IgA and IgM antibodies, as well as neutralizing antibodies against the S-protein. Cellular immunity was assessed by analyzing peripheral blood mononuclear cells (PBMC; n = 43 in first cohort, n = 32 in second cohort), including T-helper memory and cytotoxic subsets, and cytokine production after in vitro stimulation with recombinant SARS-CoV-2 proteins. A multiplex cytokine assay was used to analyze effector and regulatory immune responses. Results: Virus-specific IgG antibodies persisted for years after exposure to SARS-CoV-2, with IgG against the receptor-binding domain (RBD) correlating most strongly with neutralizing activity. Vaccinated individuals demonstrated higher IgA responses, whereas antibodies to the N-protein were associated with previous infection. No IgM antibodies were detected in any subjects, suggesting an immune response based on memory rather than ongoing infection. PBMCs from individuals with a history of both COVID-19 exposure and vaccination exhibited enhanced responsiveness, characterized by increased frequencies of memory T cells compared to vaccination alone. Stimulating with the S-protein induces higher cytokine production, including IFN-gamma, TNF-alfa, and IL-12(p70), compared with stimulation by the N-protein. Cytokines such as IL-10 and TGF-beta are also elevated, suggesting immune regulation rather than persistent inflammation. Conclusions: SARS-CoV-2 infection and vaccination are associated with persistent humoral and cellular immune responses detectable several years after exposure. Individuals with hybrid immunity exhibit broader and functionally enhanced immune reactivity, indicating more robust long-term immune memory. Future studies should focus on the long-term consequences of hybrid immunity and optimize other vaccine strategies, including recombinant antigen vaccines.

Biomedicines doi: 10.3390/biomedicines14040921

Authors: Michał Porada Bartosz Pawełczak Karolina Barańska-Pawełczak Krzysztof Marciniec

Background/Objectives: Myocardial ischemia–reperfusion injury (MIRI) remains an ever-growing threat in the field of cardiology, as it has become a major risk factor for unfavorable outcomes following reperfusion therapies. Oxidative stress and inflammation remain the key pathophysiological mechanisms underlying MIRI, and the presently available treatments fail to prevent this process effectively. This systematic review aimed to summarize and critically assess the latest preclinical research (2020–2026) on nanocarrier-based interventions targeting oxidative stress in MIRI, highlighting the potential of the new nanostructures in cardioprotection. Methods: A total of 24 studies meeting the PRISMA criteria have been found through a literature search of PubMed, Embase, and Web of Science databases published between 2020 and 2026. The studies eligible for inclusion had focused on the efficacy of nanocarrier-based interventions in preclinical studies of MIRI. Results: Of the 24 included studies, all investigated nanocarrier-based interventions in preclinical models of MIRI. In vitro, ex vivo, and in vivo models were diverse, with most studies being a combination of both in vitro and in vivo models. Commonly studied were lipid-based nanocarriers, polymeric nanoparticles, and biomimetic nanocarriers. Across studies assessed for this review, treatments with nanocarriers were seen to suppress inflammatory and oxidative stress pathways, with a few studies showing a suppression of cardiomyocyte apoptosis. Cardiac function was restored as determined by echocardiography analyses or ex vivo models of the myocardium, thus validating that the nanocarrier-mediated therapies are effective against MIRI. Conclusions: The analyzed preclinical studies indicate that the described therapies could provide a promising basis for future clinical trials in the treatment of MIRI, provided their safety and efficacy are confirmed in clinical trials.

Biomedicines doi: 10.3390/biomedicines14040922

Authors: Antonio Avelino Ferreira Soares Yago Rodrigues Gontijo Dante Mafra Tourino Teixeira Bruna Rodrigues Gontijo Alexandre Sampaio Rodrigues Pereira Larissa Sousa Silva Bonasser Caroline Ferreira Fratelli Calliandra Maria de Souza Silva Izabel Cristina Rodrigues da Silva

Introduction: Major Depressive Disorder (MDD) has been increasingly associated with inflammatory dysregulation, particularly involving tumor necrosis factor-alpha (TNF-α). Genetic polymorphisms within the TNFA promoter region have been investigated as potential modulators of depressive susceptibility, symptom expression, treatment response, and inflammatory comorbidity. However, findings remain inconsistent across populations and clinical contexts. Methods: This systematic review adhered to PRISMA 2020 guidelines and was registered in PROSPERO (CRD420251242724). Observational and interventional studies evaluating associations between TNFA polymorphisms—specifically rs1800629 (−308 G/A), rs1799724 (−857 C/T), and rs1799964 (−1031 T/C)—and MDD-related outcomes in adults were included. Data extraction and methodological quality assessment were performed independently using an adapted GRIPS framework. Results: Eleven studies met the inclusion criteria, with eight investigating MDD without cardiovascular comorbidity and three assessing cardiovascular populations. Across diverse cohorts, rs1800629 and rs1799724 did not demonstrate consistent associations with MDD susceptibility. Although isolated population-specific findings were reported, genotype and allele distributions were generally comparable between cases and controls. Rs1799724 was associated with symptom dimensions and altered TNF-α expression in two cohorts. Rs1799964 was not linked to disease occurrence but showed potential association with antidepressant response and adverse cardiovascular outcomes in patients with chronic heart failure and comorbid depression. Overall, findings were heterogeneous and influenced by population characteristics, sample size, and clinical context. Conclusions: Current evidence does not support a robust etiological role for TNFA promoter polymorphisms in major depressive disorder. These variants may exert context-dependent modulatory effects on symptom expression, treatment response, or inflammatory-cardiovascular interactions rather than serving as primary susceptibility determinants. Larger, ethnically diverse studies integrating genetic, inflammatory, and clinical data are required to clarify the contribution of inflammatory genetic variability in depressive disorders.

Biomedicines doi: 10.3390/biomedicines14040920

Authors: Akgül Arıcı Elif Akçay Seda Ocaklı Osman Demir Fikret Erdemir

Background/Objectives: Although certain established prognostic factors may occasionally fail to provide precise risk prediction in renal cell carcinoma (RCC), valosin-containing protein (VCP)/p97 has been implicated in a poor prognosis in various cancers, while its prognostic value in clear cell renal cell carcinoma (ccRCC) remains unknown. This study aimed to determine the independent prognostic value of VCP/p97 expression in ccRCC. Methods: This retrospective study included 137 ccRCC patients, and VCP/p97 expression was analyzed by immunohistochemistry and classified into either low or high expression based on the intensity of the staining in relation to the expression in endothelial cells. Results: High expression of VCP/p97 was significantly correlated with large tumor size (p < 0.001), Fuhrman nuclear grade (p = 0.003), advanced TNM stage (p < 0.001), and distant metastasis (p < 0.001). Kaplan–Meier analysis showed that the survival of patients with high expression of VCP/p97 was significantly reduced, and multivariate analysis revealed that high expression of VCP/p97 independently predicted poor survival (HR 2.09, 95% CI 1.06–4.15, p = 0.034) in addition to age, Fuhrman grade, and TNM stage. Conclusions: This study demonstrated that VCP/p97 expression, a newly identified prognostic factor, independently predicted a poor prognosis in ccRCC, and its expression may be a useful tool in identifying ccRCC patients with a poor prognosis.

Biomedicines doi: 10.3390/biomedicines14040919

Authors: Veronika Litvinenko Rose Alkhateeb Serafima Romanova Sandaara Kovalenko Vitalii Dzhabrailov Mikhail A. Popov Mikhail Slotvitsky Evgeniy G. Agafonov Vladislav V. Dontsov Sheida Frolova Dmitriy I. Zybin Dmitriy V. Shumakov Alexander Romanov Konstantin Agladze Valeriya A. Tsvelaya

Background/Objectives: Patient-derived mesenchymal stem cells (MSCs) represent a promising avenue for myocardial regeneration, yet therapeutic application remains limited by inconsistent differentiation capacity and the absence of standardized cardiogenic induction protocols. This study demonstrates a proof-of-concept for guiding patient-specific bone marrow MSCs toward a functional cardiomyocyte phenotype using paracrine signals from differentiating iPSC-derived cardiomyocytes (iPSC-CMs). Materials and Methods: MSCs were maintained in conditioned medium from a concurrent, validated iPSC-CM differentiation protocol, with evaluation via immunocytochemistry, optical mapping, and whole-cell patch-clamp recordings. Results: Differentiated MSCs acquired organized sarcomeric architecture with cross-striations and displayed spontaneous calcium oscillations with decay kinetics matching source iPSC-CMs (CaT50 ≈ 283 ms vs. 301 ms). In co-culture, MSC-derived cells exhibited synchronized calcium dynamics with iPSC-CMs, confirming functional coupling, while patch-clamp detected hallmark cardiac ion currents (INa, ICa,L, and IKv). Morphologically, MSC-CMs displayed more mature, elongated rod-like shapes. Conclusions: Although current densities indicate partial immaturity, their reproducible detection validates successful cardiomyogenic commitment. This “parallel differentiation” platform eliminates donor-specific protocol tuning, providing a streamlined, paracrine-mediated approach to generate autologous cardiomyocyte-like cells for disease modeling, pharmacological testing, and future regenerative applications.

Biomedicines doi: 10.3390/biomedicines14040918

Authors: Li Peng Yanqiu Li Xinglian Duan Jun Long Qin Ran Xiaojuan Zeng Bin Liu Duan Wang Jian Yang

Background/Objectives: Percutaneous coronary intervention (PCI) effectively restores coronary flow in acute myocardial infarction (AMI), but myocardial ischemia–reperfusion injury (MIRI) remains a major prognostic determinant. Mitochondrial open reading frame of the 12S rRNA-c (MOTS-c) has shown cardiovascular protective effects, yet its association with MIRI is unclear. This study aimed to investigate the relationship between serum MOTS-c levels and MIRI in AMI patients. Methods: Seventy-two AMI patients undergoing PCI were enrolled and divided into MIRI (n = 34) and non-MIRI (n = 38) groups. Clinical data and MOTS-c levels in peripheral serum and intracoronary blood were compared. Multivariate logistic regression and receiver operating characteristic (ROC) analysis were performed to identify MIRI predictors. Results: The MIRI group exhibited lower systolic blood pressure, preoperative thrombolysis in myocardial infarction (TIMI) grade, and HDL-C, but higher total ischemic time, door-to-balloon time, culprit vessel stenosis severity, Killip grade and adverse event incidence (all p < 0.05). Postoperative peripheral serum MOTS-c levels were significantly lower in the MIRI group than in the non-MIRI group (p < 0.05), while preoperative peripheral and intracoronary MOTS-c levels showed no significant differences between groups. Multivariate logistic regression identified postoperative peripheral MOTS-c levels (OR = 0.986, 95%CI: 0.976–0.996) and preoperative TIMI grade ≥ 1 (OR = 0.036, 95%CI: 0.004–0.309) as independent protective factors for MIRI, whereas serum creatinine was identified as an independent risk factor. ROC analysis demonstrated that postoperative peripheral MOTS-c levels predicted MIRI with an area under the curve of 0.648. Conclusions: Postoperative peripheral serum MOTS-c levels represent an independent protective factor against MIRI in patients with acute myocardial infarction and suggest a potential predictive value for MIRI, although its clinical utility as a standalone predictor requires further validation through dynamic monitoring and larger-scale studies. This finding may offer a potential novel biomarker and therapeutic direction for MIRI.

Biomedicines doi: 10.3390/biomedicines14040916

Authors: Ah Young Leem Shihwan Chang Chanho Lee Mindong Sung Hye Young Hong Geun In Lee Youngmok Park Seung Hyun Yong Sang Hoon Lee Song Yee Kim Kyung Soo Chung Eun Young Kim Ji Ye Jung Young Ae Kang Moo Suk Park Young Sam Kim Se Hyun Kwak Su Hwan Lee

Background/Objectives: Ventilator weaning imposes profound hemodynamic stress, unmasking cardiopulmonary vulnerability. Since conventional predictors of post-tracheostomy weaning failure remain elusive, biomarker-driven risk stratification offers a translational approach. We evaluated the prognostic utility of admission N-terminal pro-B-type natriuretic peptide (NT-proBNP) as an early triaging tool for weaning failure and explored its therapeutic implications alongside optimal tracheostomy timing. Methods: In this large-scale retrospective cohort study, we analyzed 707 critically ill patients who underwent tracheostomy in a medical intensive care unit. We investigated the association between baseline NT-proBNP levels—measured as a molecular surrogate of cardiovascular stress at ICU admission; echocardiographic parameters; and weaning outcomes. Multivariable logistic regression analysis was utilized to identify independent pathophysiological predictors associated with weaning failure. Results: Patients experiencing weaning failure exhibited significantly elevated admission NT-proBNP levels compared to those successfully weaned (3077.0 vs. 1410.0 pg/mL, p < 0.001). High admission NT-proBNP (>3271 pg/mL) was independently associated with an increased risk of weaning failure (adjusted odds ratio [aOR] 2.86, 95% confidence interval [CI] 1.81–4.53, p < 0.001). Conversely, an early clinical intervention—tracheostomy performed within 10 days of mechanical ventilation initiation—was associated with a significantly lower risk of weaning failure (aOR 0.55, 95% CI 0.35–0.87, p = 0.010). Furthermore, elevated biomarker levels strongly correlated with prolonged intensive care unit stays and higher 90-day mortality. Conclusions: Admission NT-proBNP serves as a powerful biomarker associated with cardiopulmonary vulnerability from the earliest stages of critical illness. Integrating this diagnostic biomarker with interventional strategies like optimal tracheostomy timing has significant prognostic implications. This biomarker-guided approach facilitates personalized risk stratification from ICU admission, potentially optimizing weaning pathways for mechanically ventilated patients.

Biomedicines doi: 10.3390/biomedicines14040915

Authors: Baran Balcan Duygu Vezir Ramazan Ocal Caner Cinar Erdal Aksoy Yeliz Celik Berrin Ceyhan

Background: Anemia and obstructive sleep apnea (OSA) are prevalent comorbidities in bronchiectasis and may share overlapping pathophysiological mechanisms. However, their combined impact on clinical outcomes in bronchiectasis remains underexplored. We aimed to investigate the associations between anemia, OSA, and clinical characteristics in patients with bronchiectasis, including cystic fibrosis (CF) and non-CF subtypes. Methods: 70 adults with bronchiectasis (35 CF-related, 35 non-CF) underwent polysomnography. Anemia was defined using standard hemoglobin < 13 g/dL for men and <12 g/dL for women. Clinical, nutritional, and sleep-related variables were assessed, and associations with anemia were evaluated. Results: Anemia was present in 24.3% of participants. Compared to non-anemic patients, those with anemia had significantly higher rates of female sex (38.5% vs. 6.5%, p = 0.002), nutritional problems (47.1% vs. 20.8%, p = 0.034), OSA prevalence (94.1% vs. 54.7%, p = 0.003), and annual hospitalizations (1.41 ± 0.41 vs. 0.43 ± 0.10, p = 0.002). In multivariate analysis, female sex (OR: 12.32; 95% CI: 3.12–45.96; p = 0.002) and OSA (OR: 4.70; 95% CI: 2.67–45.29; p = 0.007) remained independent predictors of anemia. Subgroup analysis showed a significant univariate association between OSA and anemia in CF patients (p = 0.045), while hospitalization frequency was an independent predictor of anemia in non-CF bronchiectasis (p = 0.040). Conclusions: Anemia in bronchiectasis is independently associated with female sex and OSA, with additional exploratory subgroup findings. These findings indicate an association between OSA and anemia within bronchiectasis populations; however, the cross-sectional design precludes conclusions regarding directionality or causality.

Biomedicines doi: 10.3390/biomedicines14040914

Authors: Jaroslav Pelisek Yankey Yundung Anna-Leonie Menges Fabian Rössler Benedikt Reutersberg Alexander Zimmermann Martin Geiger

Background/Objectives: Nuclear receptor corepressors NCOR1 and NCOR2 are key regulators of transcriptional repression, chromatin remodelling, and immunometabolic signalling. While NCOR1 has already been linked to vascular biology, its relevance in abdominal aortic aneurysm (AAA) remains unclear, particularly for NCOR2. This study aimed to investigate the expression, cellular localisation, and molecular interactions of NCOR1/2 in human AAA tissue. Methods: Human AAA samples (elective and ruptured) (n = 45) and non-aneurysmal control aortas (n = 18) were obtained from our Swiss Vascular Biobank. Transcriptomic profiling was performed using ribosomal RNA-depleted RNA sequencing. Differential expression and correlation analyses were performed using DESeq2/EdgeR and Spearman rank correlation with Benjamini–Hochberg correction. Cellular localisation was assessed through immunohistochemistry (IHC). Results: Bulk transcriptomic analyses showed no significant differences in NCOR1 or NCOR2 expression between AAA and controls. IHC revealed that NCOR1 was found in endothelial cells (ECs), smooth muscle cells (SMCs), and inflammatory infiltrates, while NCOR2 was primarily associated with macrophages. Correlation analyses suggest that NCOR1 interacts with various cellular markers, proteolytic enzymes, inflammatory mediators, and epigenetic regulators, including the lncRNA MALAT1. NCOR2 showed distinct associations with remodelling enzymes, TGFB1 signalling, selective epigenetic modifiers, and lncRNA H19. Conclusions: The lack of transcriptional differences in NCOR1 and NCOR2 between AAA and controls does not exclude cell-type-specific regulation or functional relevance. The specific cellular distributions and molecular associations in human AAA imply that NCOR1 and NCOR2 play non-redundant roles in vascular remodelling, inflammation, and epigenetic regulation. Our findings highlight NCOR pathways as potential modulators of AAA pathophysiology and promising targets for future therapies.

Biomedicines doi: 10.3390/biomedicines14040913

Authors: Yan Wang Xiran Liu Zerui Shan Yu Xia Yiya Weng Magdalena M. Stevanović Nenad Filipović Kai Zheng Junqing Ma

Background/Objectives: Biodegradable magnesium (Mg)-based biomaterials release Mg2+ ions during degradation and may promote vascular-related regeneration. However, their effects on lymphatic endothelial cells (LECs) and lymphangiogenesis remain unclear. This study investigated whether magnesium powder-derived ionic extracts could enhance lymphangiogenesis-related behaviors of LECs in vitro. Methods: Mg powder extracts were prepared and diluted for in vitro treatment. After viability screening, Mg (1:10), Mg (1:100), and Mg (1:1000) were selected for further analysis. LEC proliferation, migration, and tube formation were assessed, together with intracellular reactive oxygen species (ROS) levels and the expression of VEGFA, VEGFC, and VEGFR3. Results: Mg (1:10) and Mg (1:100) showed good cytocompatibility and significantly promoted LEC proliferation, migration, and tube formation compared with the control and Mg (1:1000) groups. These effects were accompanied by reduced intracellular ROS levels and increased expression of VEGFA, VEGFC, and VEGFR3. Conclusions: Magnesium powder-derived ionic extracts enhance lymphangiogenesis-related responses of LECs in vitro, particularly at the 1:10 and 1:100 dilutions. These findings support the potential of Mg-based biodegradable biomaterials for lymphatic tissue regeneration.

Biomedicines doi: 10.3390/biomedicines14040912

Authors: Yujiang Luo Yaxiong Deng Li Yuan Siqi Fu

A keloid is a benign fibroproliferative cutaneous disorder characterized by excessive extracellular matrix deposition, which is driven by persistent fibroblast proliferation and aberrant wound healing. Its complex pathogenesis involves genetic susceptibility, chronic inflammation, mechanical tension and dysregulated cellular signaling, resulting in poor clinical efficacy and high recurrence rates. Cellular senescence has recently become a central focus in exploring keloid pathophysiology, offering a novel perspective for elucidating its initiation, progression and recurrence. This review systematically summarizes the biological roles of cellular senescence in keloid pathology: it elaborates on the basic concepts and core molecular features of cellular senescence, details the spatial heterogeneity of senescent cell accumulation, the activation and pathological effects of senescence-associated secretory phenotype (SASP), and clarifies the molecular link between senescence-resumed proliferation (SRP) and keloid recurrence and treatment resistance. It also summarizes advances in senescence-related markers, the regulatory roles of the p53/p21 and Wnt/β-catenin pathways, and potential senescence-targeted therapies (senolytic, senomorphic, signaling intervention, cell reprogramming). Finally, we discuss the challenges and future perspectives for translating senescence research into clinical keloid treatments, aiming to provide a novel theoretical framework and therapeutic targets for keloid management.

Biomedicines doi: 10.3390/biomedicines14040910

Authors: Lingzhi Peng Na Sun Biqiong Ren

Cuproptosis represents a novel form of programmed cell death that relies on copper ions and targets the mitochondrial tricarboxylic acid cycle, offering fresh avenues for tumor therapy. Elesclomol, as a highly efficient small-molecule copper ion carrier, transports copper ions into mitochondria. Under the action of ferredoxin-1 (FDX1), it induces abnormal aggregation of lipoylated proteins and loss of iron–sulphur clusters, thereby generating protein toxicity stress and killing tumor cells. Furthermore, elesclomol effectively remodels the tumor immune microenvironment by promoting dendritic cell maturation and CD8+ T cell infiltration, demonstrating synergistic effects with immune checkpoint blockade therapies. However, tumor cells can develop resistance mechanisms through metabolic reprogramming via hypoxia-inducible factor-1α (HIF-1α) and the nuclear factor E2-related factor 2 (Nrf2)-driven reductive pathway, which partially limits the drug’s clinical efficacy. Addressing this limitation, combination therapies integrating elesclomol with targeted agents such as ferroptosis inducers or chemotherapeutic drugs have demonstrated significant antitumor advantages. Future research must urgently leverage the selection of precise biomarkers and the development of novel intelligent nanodelivery systems to further advance the safe and efficient clinical translation of elesclomol.

Biomedicines doi: 10.3390/biomedicines14040911

Authors: John K. Triantafillidis

Pyroptosis, a gasdermin-mediated and highly immunogenic form of regulated cell death, has surfaced as a critical determinant in the progression and therapeutic landscape of gastrointestinal (GI) cancers. Unlike non-inflammatory apoptotic pathways, pyroptosis involves the assembly of inflammasome complexes and the subsequent activation of caspases, leading to the cleavage of gasdermin proteins and the formation of transmembrane pores. It contributes to tumor suppression via immunogenic cell death and activation of antitumor immunity but may also promote tumor progression through chronic inflammation and remodeling of the tumor microenvironment. In this comprehensive review, we delineated the molecular architecture of pyroptotic signaling within the GI tract, highlighting the “double-edged sword” nature of this process. We further evaluated its role in the pathogenesis of GI cancers and in emerging translational strategies, including the pharmacological modulation of gasdermins and microbiome-based interventions, aiming to integrate pyroptosis induction into current immunotherapeutic frameworks.

Biomedicines doi: 10.3390/biomedicines14040907

Authors: Irmak Fatoş Korkmaz Tugba Elgun Çiğdem Aktas Ersin Gündeğer Asiye Gok Yurttas

Background: Colorectal cancer (CRC) is one of the most prevalent malignancies worldwide and remains a major health concern due to its high recurrence and mortality rates. Recent studies suggest that anesthetic agents, including ketamine, may have direct effects on cancer cell viability and apoptosis. Objective: This study aimed to investigate the in vitro effects of ketamine on the HT-29 human colorectal adenocarcinoma cell line, focusing on its cytotoxic and pro-apoptotic potential. Material and Methods: HT-29 cells were treated with ketamine for 24 h. Cell viability was evaluated using the MTT assay. Apoptosis rates were determined by flow cytometry with Annexin V-FITC/7-AAD staining. Furthermore, quantitative PCR (qPCR) was performed to assess the expression levels of key genes associated with proliferation and apoptosis. GeneQuery™ Human Basal Cell Carcinoma qPCR Array Kit (GQH-BCC-GK015-C) was used for qPCR analysıs. Molecular docking simulations were performed to investigate the potential molecular interactions between ketamine and three target proteins: the N-methyl-D-aspartate (NMDA) receptor, epidermal growth factor receptor (EGFR), and casein kinase 1 delta (CSNK1D). To ensure robustness of predictions, two independent docking methods were employed. Results: Ketamine significantly reduced cell viability in a dose-dependent manner, with an IC50 value of approximately 1.05 µM. Flow cytometry analysis demonstrated a marked increase in early apoptosis (23.9%) in treated cells. These findings suggest that ketamine exhibits potential anti-proliferative and pro-apoptotic effects on HT-29 colorectal cancer cells. Conclusions: These findings suggest that ketamine exhibits potential anti-proliferative and pro-apoptotic effects on HT-29 colorectal cancer cells in vitro. Further studies are warranted to elucidate the underlying molecular mechanisms and potential clinical implications.