Biomedicines doi: 10.3390/biomedicines14081742

Authors: Peter Alekseevich Shatalov Anna Aleksandrovna Bukaeva Egor Mikhailovich Veselovsky Alexey Aleksandrovich Traspov Maria Alexandrovna Revkova Polina Vladimirovna Mishakova Elena Alekseevna Kirillova Maria Pavlovna Raygorodskaya Daria Valerievna Bagdasarova Irina Alexeevna Leukhina Anna Petrovna Shinkarkina Alena Valerevna Murzaeva Yulia Arturovna Mechenici Andrei Dmitrievich Kaprin Petr Viktorovich Shegai

Background/Objectives: Triple-negative breast cancer (TNBC) is enriched for germline pathogenic variants in BRCA1/2 and other DNA repair genes, but the additional value of extended germline profiling beyond BRCA1/2 remains insufficiently characterized in Russian patients. We aimed to characterize germline pathogenic/likely pathogenic variants (GPV/LPVs) and variants of uncertain significance (VUSes) in cancer predisposition and homologous recombination repair (HRR)-related genes in Russian TNBC patients and to assess the additional yield of extended germline profiling beyond BRCA1/2. Methods: We retrospectively analyzed germline DNA from 275 patients with histologically confirmed TNBC. Exome sequencing was performed for 204 patients and focused HRR-panel testing for 71 patients on the MGISEQ-G400 platform. Results: Overall, 114 patients (41.5%) harbored at least one germline GPV/LPV. BRCA1/2 GPV/LPVs were detected in 53 patients (19.3%), with BRCA1 predominating over BRCA2 (48 vs. 5 carriers) and the recurrent BRCA1 c.5329dup variant accounting for 23 cases. In the exome subset, 79 of 204 patients (38.7%) carried a GPV/LPV in Groups 1–3, including 48 (23.5%) with BC-related GPV/LPVs; non-BRCA Group 1 genes contributed seven additional carriers beyond BRCA1/2-only analysis. The expanded HRR set identified 52 GPV/LPV carriers (25.5%) versus 45 (22.1%) in the reference HRR panel, while the common HRR gene set identified 68 carriers (24.7%) in the full cohort. In addition, 183 unique VUSes were found in Groups 1–3 in the exome subset, affecting 128 patients (62.7%). Conclusions: Russian TNBC patients show a substantial germline GPV/LPV burden dominated by BRCA1/2 alterations and the recurrent BRCA1 c.5329dup founder variant. Extended germline profiling identified additional non-BRCA and HRR-related findings beyond BRCA1/2, but the incremental yield was moderate and accompanied by a considerable VUS burden. Broader germline testing may therefore support hereditary risk assessment and exploratory HRR-focused stratification, but non-BRCA HRR findings should not be considered sufficient for therapy selection without additional tumor-level or clinical evidence.

Biomedicines doi: 10.3390/biomedicines14081741

Authors: Vlad-Alexandru Ionescu Gina Gheorghe Claudiu Stefan Turculet Teodor Florin Georgescu Razvan Matei Bratu Ioana-Alexandra Baban Cristina Mambet Carmen Cristina Diaconu Valentin Enache Laura Denisa Dragu Ana Iulia Neagu Ioana Madalina Pitica Marian Constantin Camelia Cristina Diaconu Coralia Bleotu

Background: The mechanistic link between HERV-associated transcript expression and the inflammatory pathways involved in colorectal cancer (CRC), particularly the IL-17 axis, remains insufficiently characterized. We performed an exploratory paired-tissue study to investigate whether HERV-associated transcript deregulation in CRC aligns with a distinct local inflammatory signature, potentially consistent with endogenous retroviral re-expression linked to local immune activation and viral mimicry-related processes within the tumor microenvironment. Methods: Matched colorectal tumor tissue (T) and histologically tumor-free adjacent tissue (N) were obtained from 21 treatment-naive CRC patients. Expression of selected HERV-associated loci/transcripts (HERV-FRD, HERV-R, HERV-K, HERV-V1, HERV-1, HERV-H, and ERVWE1), HHLA1/HHLA2, IL-17 family members, and NFKB1 was assessed by RT–qPCR. Results: CRC tumors exhibited selective upregulation of HERV-FRD, HERV-R, HERV-K, HERV-V1, ERVWE1, HHLA1, and HHLA2, whereas HERV-1 and HERV-H showed no significant paired differences. IL-17E was also increased in tumor tissue, while NFKB1 did not show significant paired variation. Paired differential expression analysis [Δ(T−N)] identified a coordinated HERV/ERVWE1 signature that positively correlated with IL-17E after FDR correction, whereas HHLA1/HHLA2 displayed partially distinct behavior. No consistent associations were observed with clinicopathological variables. Conclusions: CRC appears to harbor a selective HERV/ERVWE1-associated transcriptional program linked to IL-17E, supporting a candidate inflammatory axis that may reflect a biologically relevant component of the tumor microenvironment. This expression pattern may also be compatible with a localized endogenous retroviral reactivation state linked to viral mimicry-related inflammatory signaling, although this interpretation remains indirect and requires mechanistic validation. These findings are associative and hypothesis-generating, but they provide a conceptual basis for future mechanistic validation and for exploring whether HERV-linked inflammatory signatures may have biomarker or therapeutic relevance in CRC.

Biomedicines doi: 10.3390/biomedicines14081740

Authors: Stojanovic Djokic Stojanovic Stojanovic Jovanovic Matic

Eosinophils are tissue-adapted granulocytes with important roles in host defense, immune regulation, barrier responses, and inflammatory disease. Beyond classical degranulation, activated eosinophils can release eosinophil extracellular traps (EETs), extracellular DNA and protein structures that arise through cytolytic nuclear EETosis or rapid mitochondrial DNA (mtDNA) release. These traps may contribute to host protection by immobilizing pathogens and concentrating eosinophil granule proteins at sites of mucosal or tissue inflammation. However, when EET formation is excessive, persistent, or insufficiently cleared, the same structures may promote epithelial injury, mucus viscosity, thromboinflammation, tissue remodeling, fibrosis, and chronic eosinophilic inflammation. This review summarizes the cellular mechanisms, structural organization, inducing stimuli, and disease relevance of EETs. Particular attention is given to infectious settings, eosinophilic airway and middle-ear disease, systemic vasculitic and hypereosinophilic disorders, inflammatory skin and ocular diseases, eosinophilic gastrointestinal disease, and cancer. Current evidence indicates that EETs should be interpreted as context-dependent eosinophil effector structures with dual biological potential. Although their mechanistic relevance is increasingly recognized, further histological, functional, and clinical validation is needed before EETs can be established as reliable biomarkers or therapeutic targets.

Biomedicines doi: 10.3390/biomedicines14081738

Authors: Shiyun Peng Zhenkun Wen Xiao-Ting Cai

Background/Objectives: Glioblastoma (GBM) is widely recognised as a highly aggressive form of malignant tumour arising within the central nervous system. The proneural-to-mesenchymal (PN–MES) state transition is a key process underlying malignant progression and therapeutic resistance. The present study was designed to elucidate the association between the PN–MES transition and arachidonic acid (AA) metabolic reprogramming in GBM. Methods: We integrated four public single-cell RNA-sequencing cohorts, two spatial transcriptomic cohorts, and three bulk RNA-sequencing cohorts for GBM. By combining single-cell transcriptomics, spatial transcriptomics, pseudotime trajectory inference, and cell–cell communication analysis, we systematically evaluated the cell-state dependence and spatial heterogeneity of AA metabolism in GBM. We further incorporated machine learning-based survival modelling, molecular docking, molecular dynamics simulations, and in vitro functional assays to identify and validate potential prognostic biomarkers and therapeutic targets. Results: Mesenchymal-like (MES-like) tumour cells showed the highest transcriptionally inferred AA metabolism-related score, and AA metabolism-related gene expression was closely coupled with PN–MES state transition. The peptidylprolyl isomerase A (PPIA)–basigin (BSG) signalling axis was selectively enriched in tumour cells with high expression of arachidonic acid metabolism-related genes (AAMGs), whereas virtual knockout of BSG perturbed MES- and invasion-related gene modules. Spatial transcriptomic analysis confirmed that PPIA–BSG-associated communication was enhanced within MES-like tumour niches and was coupled with inflammatory and hypoxic programmes. AA metabolism-related genes enabled robust prognostic stratification. Molecular simulation and in vitro experiments suggested that Venetoclax could bind BSG and suppress GBM cell viability. Conclusions: AA metabolism-related transcriptional reprogramming defines mesenchymal-associated malignant niches in GBM and may contribute to PN–MES-related state evolution through PPIA–BSG-mediated microenvironmental communication. AA metabolism-related features and the BSG-associated signalling axis may provide candidate directions for prognostic stratification and targeted intervention.

Biomedicines doi: 10.3390/biomedicines14081739

Authors: Shreeram Behera Koushik Bhattacharya Soumya Jal Gopal Krishna Purohit Ashok Kumar Sah Shagun Agarwal Rasha Babiker Ashwani Bhardwaj Ayman Husein Mohamed Alfeel

Metabolic syndrome (MetS) is a complex metabolic disorder that includes central fat accumulation and insulin resistance, abnormal lipid levels, high blood pressure, and ongoing mild inflammation as its main symptoms. The global rise in metabolic syndrome parallels the increasing prevalence of male infertility, suggesting a potential association between metabolic health and reproductive function. Male fertility suffers from obesity, which represents a major element of metabolic syndrome because it disrupts hormonal balance, raises scrotal temperatures, and causes oxidative damage and HPG axis malfunction. Excess body fat functions as an endocrine system that induces the body to produce pro-inflammatory cytokines, which include tumor necrosis factor-α and interleukin-6, and various other inflammatory substances that result in ongoing body-wide inflammation. The state of inflammation leads to damaged spermatogenesis, which results in lower testosterone production and impaired semen quality that includes changes in sperm concentration, motility, morphology, and DNA integrity. The combination of insulin resistance and metabolic disturbances that occur in MetS leads to oxidative stress and mitochondrial dysfunction, which harms male reproductive abilities in testicular tissue. Recent research shows that adipokines and endocrine disruptors, together with epigenetic modifications, function as mediators that establish the connection between metabolic syndrome and male infertility. The development of targeted therapeutic strategies and lifestyle interventions requires researchers to study how obesity and chronic inflammation interact with each other to create reproductive dysfunction. This review defines the pathophysiological mechanisms that connect metabolic syndrome to male infertility by focusing on how obesity-related inflammation affects male reproductive health.

Biomedicines doi: 10.3390/biomedicines14081737

Authors: Ángela Y. García Fonseca Carlos Javier Alméciga-Díaz Andrés F. Aristizábal-Pachón

Background: Metastasis is the leading cause of mortality in lung cancer and is regulated by multiple molecular mechanisms, including microRNAs (miRNAs). Although cancer stem cells (CSCs) and epithelial–mesenchymal transition (EMT) contribute to metastatic progression, the miRNA networks underlying these phenotypes remain poorly characterized in non-small cell lung cancer (NSCLC). Aim: To identify miRNA signatures associated with CSCs and EMT in NSCLC and functionally characterize hsa-let-7a-3p. Methods: EMT was induced in A549 and NCI-H1975 cells by dCas9-mediated activation of TWIST, whereas CSC-enriched populations were generated by CD133-based sorting and stem cell culture conditions. Small RNA sequencing, bioinformatic analyses, qPCR validation, and functional assays were performed to identify and characterize phenotype-associated miRNAs. Results: Small RNA sequencing identified distinct miRNA expression profiles associated with EMT and CSC enrichment. Comparative analysis identified 13 commonly downregulated and 13 commonly upregulated miRNAs shared by CSCs from A549 and H1975 cells, suggesting conserved post-transcriptional regulatory mechanisms. Functional enrichment and miRNA–target interaction network analyses linked the miRNA signatures to pathways involved in epithelial plasticity, stemness, and tumor progression, including Wnt, TGF-β, mTOR, focal adhesion, adherens junction, and regulation of the actin cytoskeleton. Among the dysregulated miRNAs, hsa-let-7a-3p was consistently upregulated in CD133+ CSC-enriched cells from both NSCLC cell lines. Functional assays showed that hsa-let-7a-3p overexpression significantly reduced clonogenic capacity and showed a trend toward decreased invasion without affecting proliferation. Conclusion: This study identifies miRNA signatures associated with CSC-enriched and EMT-associated phenotypes in NSCLC and demonstrates that these signatures represent coordinated post-transcriptional regulatory programs involved in epithelial plasticity, stemness, and metastatic progression. Functional validation of hsa-let-7a-3p further supports its role as a context-dependent regulator of CSC biology and highlights the potential of miRNA signatures as diagnostic biomarkers in NSCLC.

Biomedicines doi: 10.3390/biomedicines14081736

Authors: Victor Pasecinic Dorin Novacescu Flavia Zara Cristina-Stefania Dumitru Antonia Armega-Anghelescu Silviu Latcu Alin Cumpanas Radu Caprariu Pavel Banov Ademir Horia Stana Radu Gheorghe Dan Raluca Dumache

Background/Objectives: International Society of Urological Pathology (ISUP) Grade Group discordance between prostate biopsy and radical prostatectomy (RP) affects 25–50% of men with localized prostate cancer and has direct implications for active surveillance, nerve-sparing, and adjuvant-treatment decisions. We synthesized contemporary evidence on the frequency of biopsy-to-RP grade migration and on emerging imaging, molecular/genomic, and artificial-intelligence (AI)-based predictors of migration. Methods: MEDLINE (PubMed) was searched from 1 January 2010 to 5 May 2026, supplemented by citation searching of recent systematic reviews. Eligible studies reported paired biopsy–RP pathology under the modified 2005 Gleason or the 2014/2019 ISUP Grade Group system, with ≥50 paired cases. Risk of bias was assessed with QUIPS, PROBAST, and QUADAS-2; certainty of evidence was rated with a GRADE framework adapted for prognostic research. Synthesis followed the SWiM reporting guidance. Results: Thirty-seven primary studies and eight prior systematic reviews were included. Concordance ranged from 44% to ~70%, upgrading from 14% to 67%, and downgrading from 5% to 26%. Biopsy GG1 disease showed the highest absolute upgrading risk (55–67%). Four predictors reached moderate GRADE certainty: PI-RADS category, biopsy approach (combined vs. systematic), PSMA-PET maximum standardized uptake value, and PSA density (PSAD). Cribriform/intraductal carcinoma at biopsy, the Decipher genomic classifier, the Prostate Health Index, and machine-learning and radiomics models reached very low certainty; clinical nomograms reached low certainty; germline alterations as direct predictors reached very low certainty. PROBAST flagged the analysis domain as high risk in five of eight prediction-model studies, and only one of eight models was externally validated. Conclusions: Despite a decade of refinements in grading, imaging, biopsy technique, and molecular profiling, biopsy-to-RP grade discordance remains substantial. Contemporary evidence supports incorporating PI-RADS, biopsy approach, and PSAD into shared decision-making; emerging molecular and AI-based predictors require prospective external validation in adequately powered, geographically representative cohorts before routine clinical adoption.

Biomedicines doi: 10.3390/biomedicines14081735

Authors: Hui Li Junyu Zhou Xu Zhu

Background: Hepatocellular carcinoma (HCC) is the predominant histological subtype of primary liver cancer, and many patients with unresectable disease require systemic therapy. This narrative review asks how current systemic regimens should be selected and sequenced, how oncogenic pathways inform resistance and biomarker development, and which emerging platforms have sufficient evidence for clinical translation. Methods: We synthesized key oncogenic pathways, pivotal trials, major guidelines, and selected translational studies cited in this review. Randomized phase III evidence was prioritized for clinical claims, whereas observational, early-phase, and preclinical studies were used to define evidence gaps and emerging approaches. Results: Immune-based combinations have reshaped first-line treatment for advanced HCC. Treatment selection must account for liver function, performance status, bleeding risk, and contraindications to anti-VEGF or immune checkpoint therapy. Evidence for post-immunotherapy sequencing remains largely observational, and validated predictive biomarkers are scarce. Conclusions: Future progress requires prospective sequencing studies, biomarker-enriched trials, and rigorous early-phase evaluation of cell, antibody, gene-editing, viral, and nanodelivery platforms.

Biomedicines doi: 10.3390/biomedicines14081734

Authors: Galya Popova-Daskalova Kostadin Kostadinov Krasimira Eneva Albena Masarlieva Dimitar Vuchev Angel Uchikov Lyubka Aleksova Nikolay Nikolov Tanya Nedkova

Background/Objectives: Cystic echinococcosis is primarily managed surgically, yet postoperative recurrence affects 2–22% of patients despite albendazole prophylaxis. This study describes the tolerability of adding praziquantel to postoperative albendazole and reports recurrence signals where efficacy cannot be inferred. Methods: In this single-centre, retrospective, two-arm cohort, 60 patients undergoing hydatid cyst surgery (2024–2026) received albendazole and 30 additionally received praziquantel. The follow-up was substantially shorter in the combination arm (median 10 versus 24 months), and recurrence was first ascertainable only from 12 months, the central limitation. Analyses used standardized mean differences, person time incidence, propensity score and inverse probability weighting, a DAG-based hepatotoxicity model, and a Bayesian Beta-Binomial re-analysis. Results: Baseline covariates were markedly imbalanced (14/15 with |SMD| > 0.10). Two recurrences occurred, both in the albendazole arm (3.73/100 person-years) and none in the combination arm’s shorter window. Adverse events were similar (43.3% vs. 50.0%), with no added hepatotoxicity after adjustment (odds ratio 1.45, 95% CI 0.49–4.37). The Bayesian posterior probability of benefit (0.88–0.93) remained below 0.95, a descriptive signal only. Conclusions: Albendazole plus praziquantel was well tolerated, but severe baseline imbalance, differential follow-up, and only two events mean efficacy cannot be assessed. The results of our study are hypothesis-generating. Further confirmation requires a prospective, multicentre randomized trial with balanced long-term follow-up.

Biomedicines doi: 10.3390/biomedicines14081733

Authors: Mete Hakan Karalök Bağnu Dündar Ayhan Parmaksız Tugba Elgun Gül Ipek Gündogan Asiye Gök Yurttaş

Background: Anti-Müllerian hormone (AMH) is widely used as a marker of ovarian reserve and is strongly influenced by chronological age. Although inflammation has been implicated in ovarian aging and follicular dysfunction, whether routinely measured peripheral inflammatory markers provide additional information regarding AMH concentrations remains unclear. Objective: This study aimed to evaluate the age-adjusted associations of C-reactive protein (CRP) and the neutrophil-to-lymphocyte ratio (NLR) with serum AMH concentrations in reproductive-age women. Methods: This retrospective cross-sectional study included women aged 18–45 years with available AMH, complete blood count, and CRP measurements. After reapplying the prespecified eligibility criteria and excluding confirmed data-entry or analytical errors, 819 women were included in the final analysis. NLR was calculated from absolute neutrophil and lymphocyte counts. Because CRP and NLR showed right-skewed distributions, Box–Cox transformations were applied. Associations with Box–Cox-transformed AMH were evaluated using an age-adjusted left-censored Tobit regression model. Results: The mean age of the participants was 34.07 ± 6.94 years, and the median AMH concentration was 1.07 ng/mL (interquartile range, 0.29–2.50). Chronological age was inversely associated with AMH (β = −0.123, 95% CI: −0.135 to −0.111, p < 0.001). After adjustment for age, neither CRP (β = −0.038, 95% CI: −0.085 to 0.009, p = 0.114) nor NLR (β = −0.012, 95% CI: −0.222 to 0.198, p = 0.912) was independently associated with AMH. Conclusions: In this retrospective outpatient cohort, routine peripheral inflammatory markers did not explain additional variation in AMH beyond chronological age. These results do not exclude a potential role for local ovarian inflammation, which may not be adequately captured by peripheral CRP or NLR measurements.

Biomedicines doi: 10.3390/biomedicines14081732

Authors: Xi-Feng Jin Ling Liu Jun-Jun Zhang

Resistance to mTOR inhibition is mediated by multiple adaptive mechanisms, including feedback activation of the PI3K/AKT/mTOR pathway, autophagic adaptation, metabolic reprogramming, immune evasion, tumor heterogeneity, and microenvironmental crosstalk. Recent research has also elucidated the crucial role of RNA modifications, particularly m6A methylation, and the regulatory impact of non-coding RNAs in facilitating tumor adaptation to mTOR blockade. Accordingly, novel therapeutic approaches are emerging. The combination of PI3K/AKT/mTOR inhibitors with immune checkpoint blockades represents a promising strategy for overcoming therapeutic resistance. Additionally, next-generation mTOR inhibitors and synthetic lethality strategies are being tailored to target specific tumor profiles. Furthermore, advancements in multi-omics technologies and AI-powered predictive models are transforming personalized cancer treatment. This comprehensive review delves into the molecular intricacies of mTOR inhibitor resistance and explores innovative strategies aimed at enhancing therapeutic outcomes and improving patient responses.

Biomedicines doi: 10.3390/biomedicines14081731

Authors: Ismael Leyva Martínez Abdi Ramírez Arteaga Hugo Martínez-Rojano Víctor Arturo Rocha Herrera Josué Salvador Aguilar Aja

Background and Objective: Pulsed electromagnetic fields (PEMF) and extracorporeal magnetotransduction therapy (EMTT) have shown promising results in various musculoskeletal disorders; however, substantial heterogeneity in application parameters has hindered the establishment of clear relationships between electromagnetic dosing and clinical response. This systematic review aims to evaluate the safety and effectiveness of PEMF and EMTT therapies for treating chronic musculoskeletal injuries in adults, based on research from 2015 to 2026. By analyzing how specific dosage parameters (such as intensity, frequency, and duration) influence clinical results, the study seeks to establish standardized, evidence-based guidelines for electromagnetic treatment prescriptions. Methods: A systematic review was conducted in accordance with PRISMA 2020 guidelines and prospectively registered in PROSPERO (CRD420251059359). A comprehensive search was performed in PubMed/MEDLINE, CENTRAL, and ScienceDirect for the 2015–2025 period. Randomized clinical trials, quasi-experimental studies, and case reports were included. Study selection, data extraction, and risk of bias assessment were performed by two independent reviewers using RoB 2, ROBINS-I, and Joanna Briggs Institute checklists. Results were synthesized narratively following SWiM (Synthesis Without Meta-analysis) recommendations. Results: Twenty-eight studies (1060 participants) were included. A dosimetric dichotomy emerged: high-intensity protocols with rapid field variation rates (e.g., EMTT > 60 kT/s) were consistently associated with robust improvements in structural and mechanically driven pathologies. Conversely, low-intensity protocols (e.g., <5 mT) often yielded results indistinguishable from placebo or active controls, particularly in nociplastic pain conditions. Quantitative analysis indicated that in structural pathologies, functional gains were maintained or amplified post-treatment, whereas nociplastic conditions showed potential symptom recurrence. No serious adverse events were reported, confirming a favorable safety profile. Conclusions: PEMF and EMTT are promising adjunctive therapies for musculoskeletal disorders, with efficacy appearing sensitive to electromagnetic dosimetry. However, significant methodological heterogeneity and the use of combination therapies limit these findings to exploratory trends. The lack of standardized safety reporting and technical parameters (e.g., waveform, dB/dt) underscores a critical translational gap. Future research must prioritize rigorous, standardized randomized controlled trials to establish definitive, evidence-based therapeutic recommendations.

Biomedicines doi: 10.3390/biomedicines14081730

Authors: Christoph Lange Elena Kaschina

Aneurysms represent a life-threatening disease due to their risk of rupture. Recent research points towards a role of endothelial–mesenchymal transition (EndMT) in their pathophysiology. EndMT acts as a cellular program during which endothelial cells undergo phenotypic and functional transformation into cells with mesenchymal properties. EndMT is implicated in embryogenesis, tissue repair, remodeling, angiogenesis and fibrosis. The studies performed in the last decade have gained new insights in the cell signaling pathways during aneurysm development in the context of EndMT and suggested new drug targets to prevent aneurysmal growth. This review summarizes the current knowledge on EndMT in various forms of aneurysm, such as abdominal and thoracic aortic aneurysms, intracranial aneurysms and coronary artery aneurysm due to Kawasaki disease.

Biomedicines doi: 10.3390/biomedicines14081729

Authors: Sukanta Jash John M. Sedivy

The human placenta is a temporary organ structured to optimize exchange between the maternal and fetal circulatory systems. Its fetal component consists of highly branched chorionic villi, which are anchored to the maternal uterine wall and project into the intervillous space. The outer surface of these villi is lined by a multinucleated, continuous layer called the syncytiotrophoblast, which is supported by an underlying layer of proliferative cytotrophoblast cells and the invasive extravillous trophoblast (EVT). This cellular bilayer forms a selective barrier that directly bathes in maternal blood, allowing for the efficient transfer of oxygen and nutrients while structurally preventing the direct mixing of maternal and fetal blood cells. Human placental studies have been stymied by ethical and accessibility constraints. Stem cell biology has now revolutionized the capacity to model human placental development, in particular with the derivation of human trophoblast stem cells (hTSCs) and organoids. Authentic, self-renewing human trophoblast stem cells (hTSCs) were first derived not from pluripotent stem cells but from primary tissue—first-trimester villous cytotrophoblasts and blastocysts. Derivation from human pluripotent stem cells (PSCs) followed only subsequently, along two principal routes: conversion of naive PSCs, which retain extraembryonic competence, and induction from primed PSCs, as well as by direct reprogramming of somatic cells to induced hTSCs. An important advance underlying these improvements is the mapping of a global reprogramming roadmap. Multi-omic and lineage-tracing experiments have mapped the stepwise transcriptional and epigenetic conversions of fibroblasts to hTSCs, including sequential chromatin reconfiguration, trophoblast gene network activation, and repression of somatic signatures. These results identify major regulatory bottlenecks and intermediate states, improving reprogramming fidelity. The derivation of stem-cell-based trophoblast organoids now enables complex modeling of placental architecture, function, and disease susceptibility in vitro. These organoids accurately recapitulate placental barrier functions and immunological features, allowing for examinations of maternal–fetal health, pregnancy disorders, and placental infection response to viruses like cytomegalovirus and SARS-CoV-2. Looking ahead, the integration of reprogramming and organoid technologies will propel patient-specific and tailor-made models for personalized diagnostics, drug screening, and mechanism studies. As we unravel the molecular ballet of trophoblast induction, such discoveries have the potential to bridge basic translational gaps in reproductive biology and maternal–fetal medicine.

Biomedicines doi: 10.3390/biomedicines14081728

Authors: Łukasz Rodzeń Damian Dyńka Mateusz Rodzeń Hanna Karakuła-Juchnowicz Dorota Łojko Sebastian Kraszewski Żaneta Grzywacz Benjamin Bikman Jen Unwin David Unwin Serafino Fazio

Hypertension (HTN) is one of the greatest public health challenges of the 21st century. Its prevalence has reached alarming levels in recent decades. The search for effective prevention and treatment strategies includes lifestyle choices, such as dietary interventions. Although not applicable to everyone, particularly interesting in this context is the ketogenic diet (KD), known to clinicians and also applied in epilepsy treatment for over a century. Its possible beneficial effects are increasingly often reported in many other conditions, including HTN. The aim of the present study is to analyse the effect of a KD on blood pressure, and the mechanisms that may modulate that effect. Based on the available literature, key potential pathways of the influence of a KD on blood pressure regulation have been identified: (1) reduced body weight; (2) reduced visceral adipose tissue; (3) improved insulin sensitivity; (4) improved water and electrolyte balance; and (5) anti-inflammatory effect. Meta-analyses and randomised controlled trials consistently indicate that ketogenic dietary interventions are associated with reductions in blood pressure, although the magnitude of the effect varies considerably depending on the ketogenic diet model, study population, and comparator. In light of these observations, it has been found that in patients undergoing pharmacological treatment, it may be necessary to appropriately reduce antihypertensive medication dosage in advance. To maximise the hypotensive effect of a KD, the need to ensure an adequate supply of potassium, magnesium, and high-quality products, as well as proper hydration has been emphasised. Further studies are needed, with the effect of a KD on systolic and diastolic blood pressure as the primary endpoint, taking into account the role of the qualitative composition of the diet in the observed effects.

Biomedicines doi: 10.3390/biomedicines14081727

Authors: Donghua Yu Jing Wang Mingxia Xie Youyuan Lu Hanqing Wang

Background: Bovis Calculus (NBC) is widely used for the management of convulsive disorder, but its clinical application is constrained by limited availability and high cost. Bovis Calculus Artifactus (BCA) and Bovis Calculus Sativus (BCS) are commonly employed as substitutes. However, systematic evidence comparing their anticonvulsant efficacy and pharmacological characteristics remains limited. Methods: This study evaluated the substitution potential of BCA and BCS relative to NBC and characterized their distinct pharmacological profiles in a pentylenetetrazol (PTZ)-induced acute seizure mouse model. Seizure latency and duration were evaluated, hippocampal pathology was examined by hematoxylin–eosin and Nissl staining, neurotransmitters and neuroinflammatory factors were quantified. Network pharmacology and molecular docking analyses were used to explore the underlying mechanisms. Results: The results showed that preventive administration of NBC, BCA, and BCS significantly alleviated PTZ-induced acute seizures, although their efficacy patterns differed markedly. NBC exhibited the broadest anticonvulsant spectrum, significantly prolonging seizure latency at multiple seizure stages. BCS showed a more restricted effect, primarily delaying stage IV seizures, whereas BCA predominantly reduced seizure frequency without significantly affecting seizure latency, suggesting that seizure frequency may represent a sensitive endpoint for evaluating certain anticonvulsant agents. Histopathological analyses demonstrated that all three preparations exerted neuroprotective effects in hippocampal subregions. NBC provided the most extensive morphological preservation in H&E staining, while all treatments significantly improved neuronal survival as assessed by Nissl staining. Biochemical analyses revealed distinct regulatory characteristics among the three Bovis Calculus sources. All treatments partially restored PTZ-induced neurotransmitter disturbances and attenuated neuroinflammatory responses. BCA showed prominent regulation of neurotransmitters and IL-1β, NBC exhibited broader modulation of neurotransmitter homeostasis, whereas BCS displayed stronger suppression of IL-6 and TNF-α despite limited effects on 5-HT regulation. These findings indicate that comparable protective effects against PTZ-induced acute seizures may be achieved through different neurochemical and inflammatory regulatory patterns. Network pharmacology and molecular docking analyses further suggested the potential involvement of PI3K-AKT and MAPK-related signaling pathways in the effects of NBC and its substitutes. Conclusions: Collectively, this study provides a comparative evaluation of NBC, BCA, and BCS and indicates that their protective activities against PTZ-induced seizures are associated with distinct neurochemical and anti-inflammatory profiles. These findings offer experimental evidence supporting the rational evaluation and application of alternative Bovis Calculus resources.

Biomedicines doi: 10.3390/biomedicines14081726

Authors: Jiwon Kim Taewoo Kim Han Gyeol Kim Seung Hyeun Lee

Background/Objective: The incidence and clinical utility of pleural tissue debris, occasionally encountered in drainage systems, have not been systematically evaluated. We aimed to determine the frequency of tissue debris detection in patients undergoing drainage for pleural effusion, and its utility for the differential diagnosis of exudative effusion and pathological diagnosis of malignant pleural effusion (MPE). Methods: This retrospective proof-of-concept analysis included 325 consecutive patients who underwent percutaneous effusion drainage. The incidence, histopathological findings, and potential predictors of tissue debris were evaluated. For patients with MPE, the adequacy of tissue for immunohistochemistry and molecular testing was evaluated, and the diagnostic sensitivity of tissue debris for MPE was calculated and compared with that of cytology with cell blocks. Results: Tissue debris was detected in 151 patients (46.5%), more frequently in exudates than in transudates (51.7% vs. 23.3%, p < 0.001), and most frequently in patients with MPE (64.6%, p < 0.001). MPE was the only independent predictor of tissue debris detection (adjusted odds ratio 5.74, 95% CI 2.25–9.31). The pathological findings of the debris were classified into metastatic carcinoma/lymphoma (49.0%), acute/chronic inflammation (30.4%), fibrinoid material with inflammation (17.2%), mesothelial reaction (2.6%), and acute suppurative pleuritis/abscess (0.7%). Among the 74 cases with MPE diagnosed using tissue debris, immunohistochemistry and molecular testing were feasible in 85.1% and 69.0% cases, respectively. Diagnostic sensitivity of tissue debris for MPE was significantly higher than that of cytology (77.9% vs. 66.3%; p = 0.035) with a combined sensitivity of 84.2%. Conclusions: Systematic collection of tissue debris, a previously underrecognized clinical specimen obtained during effusion drainage, may be highly valuable for the diagnosis of MPE and adequate for immunohistochemistry and molecular testing, reducing the reliance on invasive biopsy and supporting precise oncological decisions.

Biomedicines doi: 10.3390/biomedicines14081725

Authors: Jinan Abugharsa Özlem Balcıoğlu Barçın Özcem Mahmut Cerkez Ergoren Aya Badeea Ismail Selma Yilmaz

Background/Objectives: Chronic venous insufficiency (CVI) is a progressive vascular disorder characterized by sustained venous hypertension leading to structural and functional alterations of the venous wall. Despite its clinical significance, the molecular mechanisms driving venous remodeling remain incompletely understood. The Wnt signaling pathway has emerged as an important regulator of vascular homeostasis and remodeling; however, the expression and coordination of its core components in chronic venous insufficiency (CVI)-affected tissue have not been directly investigated. This study aimed to analyze mRNA expression of FZD2, DVL2, and AXIN1 in great saphenous vein specimens. In addition to gene expression analysis at the individual level, an investigation of co-regulatory networks was also performed to provide insight into Wnt cascade regulation in the varicose and healthy vein tissues. Methods: This single-center, retrospective observational study was conducted on 60 great saphenous vein specimens (32 CVI patients and 28 healthy controls) using re-al-time quantitative PCR. Results: The findings indicate that individual mRNA expression levels of FZD2, DVL2, and AXIN1 remained stable between varicose vein and healthy control tissues, but there were significant changes in transcriptional coordination among these genes in varicose vein tissues, especially enhanced co-expression between AXIN1 and DVL2. All three co-regulated genes exhibited moderate and generally comparable positive correlations in healthy controls: FZD2 with DVL2 (r = 0.526, p-value = 0.0069), DVL2 with AXIN1 (r = 0.486, p-value = 0.0161), and AXIN1 with FZD2 (r = 0.593, p-value = 0.0014). While in varicose vein tissues, DVL2-AXIN1 showed the highest correlation (r = 0.722, p-value < 0.0001), FZD2-DVL2 (r = 0.517, p-value = 0.0096) and AXIN1-FZD2 (r = 0.480, p-value = 0.0084) remained comparable to the healthy tissues. Conclusions: This finding indicates that the involvement of FZD2, DVL2, and AXIN1 in CVI may be driven by altered transcriptional coordination rather than by differential individual gene expression, which may contribute to the molecular pathology of chronic venous insufficiency.

Biomedicines doi: 10.3390/biomedicines14081723

Authors: Silviu Latcu Flaviu Lucian Petrica Dorin Novacescu Roland Costica Vlad Dema Victor Pasecinic Ionel Chiriac Mihai Baghina Radu Caprariu Radu Dragomir Razvan Bobora Alin Cumpanas

Prostate biopsy is among the most frequently performed diagnostic procedures in men, and two shifts have reshaped it over the past decade: the incorporation of multiparametric MRI (mpMRI) with targeted sampling, and migration from the transrectal to the transperineal route. Because these changes are usually adopted together, the practical choice facing many units is between a contemporary transperineal MRI-targeted pathway and the historical transrectal systematic one. This narrative review weights cancer detection, procedure-related harms, and implementation approximately equally, drawing on evidence that isolates route from targeting wherever possible. Across randomized trials and meta-analyses, the two pathways achieve broadly comparable detection of clinically significant cancer (ISUP grade group ≥ 2), although the transperineal route samples anterior and apical tumors more effectively and the largest randomized comparison reported a modest transperineal advantage. The principal transperineal benefit is a marked reduction in infectious complications, enabling reduced antibiotic prophylaxis and improved stewardship; its main cost is greater procedural discomfort. We review mpMRI and PI-RADS with illustrative examples, fusion technique, cost-effectiveness, special populations, emerging adjuncts, and divergent guidelines, concluding that route and targeting decisions are best individualized, with infection risk and lesion location dominant.

Biomedicines doi: 10.3390/biomedicines14081724

Authors: Xinyi Wang Zhouyang Qian Zhenchao Gong Lu Sun Liang Feng Yanjun Yang Xiaobin Jia

Background/Objectives: Baicalin (BA), a natural flavonoid with anti-inflammatory activity, shows promise for treating acute pharyngitis (AP) but its clinical application is hindered by poor water solubility and low oral bioavailability. Based on the clinically validated traditional Chinese medicine formula Pudilan Oral Liquid, we identified that BA and chlorogenic acid (CGA) can self-assemble into nanocomplexes (BA-CGA@NPs). This study aims to construct such a nanocomplex to enhance BA absorption and anti-AP efficacy with favorable biocompatibility. Methods: BA-CGA@NPs were prepared via supramolecular self-assembly and characterized by dynamic light scattering (DLS), X-ray diffraction (XRD), transmission electron microscopy (TEM), Fourier-transform infrared spectroscopy (FT-IR), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), and molecular dynamics simulation (MDS). In vivo pharmacokinetics evaluated BA absorption. Biocompatibility and therapeutic efficacy were assessed using lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages and an AP rat model. Results: BA-CGA@NPs were successfully formed with enhanced biocompatibility. In vitro, they reduced nitric oxide (NO), reactive oxygen species (ROS), tumor necrosis factor-alpha (TNF-α), and interleukin-1 beta (IL-1β) in macrophages. In AP rats, oral BA-CGA@NPs significantly increased systemic BA exposure, ameliorated pharyngeal histopathology, and lowered IL-1β, TNF-α, and interleukin-6 (IL-6) in serum and pharyngeal tissue, outperforming free BA or CGA alone. Mechanistic studies suggested that the anti-inflammatory effect was associated with modulation of the Toll-like receptor 4 (TLR4)/MyD88/Nuclear factor-κB (NF-κB) signaling pathway. Conclusions: Self-assembled BA-CGA@NPs enhance BA absorption and biocompatibility, alleviating AP inflammation through mechanisms associated with modulation of the TLR4/MyD88/NF-κB pathway, offering a promising nano-traditional Chinese medicine strategy for poorly soluble active ingredients.

Biomedicines doi: 10.3390/biomedicines14081721

Authors: Eman H. Yousef Mohamad A. El-Gammal Muhammed M. Salahuddin Mahmoud Abdelbadie Salem Amal Abdullah Alrashidi Ahmed G. Abd Elhameed

Background: Doxorubicin-induced cardiotoxicity (DIC) is a major limitation of anthracycline chemotherapy and is characterized by oxidative stress, apoptosis, and myocardial remodeling. Dysregulation of AMP-activated protein kinase (AMPK) and its downstream effector sirtuin-1 (SIRT1) has been implicated in the molecular pathogenesis of DIC. Vardenafil (Var), a selective phosphodiesterase-5 inhibitor, has shown cardiovascular benefits; however, its impact on AMPK/SIRT1 signaling during DIC remains unclear. Methods: DIC was induced in rats by cumulative doxorubicin administration. Cardiac function, serum injury biomarkers, oxidative stress indices, histopathological alterations, apoptosis, fibrosis, and molecular markers associated with the AMPK/SIRT1 pathway were evaluated. In addition, molecular docking was performed to assess the potential interaction of Var with AMPK. Results: Var significantly improved cardiac function and reduced serum levels of lactate dehydrogenase, creatine kinase, and blood urea nitrogen. Treatment attenuated myocardial oxidative stress, restored glutathione content, reduced lipid peroxidation, and alleviated histopathological damage. Furthermore, Var suppressed caspase-3 and TGF-β1 expression while enhancing HO-1 levels. Var treatment was associated with restoration of cardiac phosphorylated AMPK (p-AMPK) expression and increased SIRT1, Nrf2, and PPARγ levels in doxorubicin-treated rats, consistent with modulation of an AMPK/SIRT1-associated cytoprotective network. Molecular docking demonstrated favorable interactions between Var and AMPK, providing supportive in silico evidence for the observed molecular findings. Conclusions: These findings demonstrate that Var attenuates doxorubicin-induced cardiotoxicity, and its cardioprotective effects are associated with restoration of the p-AMPK/SIRT1/Nrf2/PPARγ signaling pathway, together with reductions in oxidative stress, apoptosis, and fibrosis. Collectively, these findings suggest that Var attenuates early Dox-induced cardiac injury, potentially through modulation of the AMPK/SIRT1 signaling pathway, warranting further validation in long-term and dose–response studies.

Biomedicines doi: 10.3390/biomedicines14081722

Authors: Mia Yang Ang Li Chen Lanni Song Leonard Lipovich Siew Woh Choo

Background/Objectives: Precision medicine requires translation of genetic and molecular variation into clinically actionable therapeutic targets. However, many disease-associated signals identified by genome-wide association studies (GWAS) reside in non-coding regulatory regions, making biological interpretation and therapeutic prioritization difficult. Long non-coding RNAs (lncRNAs) are regulatory molecules with growing relevance to disease mechanisms, biomarker discovery, patient stratification, and RNA-based therapeutics. This review presents a translational framework for AI-guided lncRNA target discovery, linking non-coding genetic signals to experimental validation and clinical implementation. Methods: This narrative review synthesizes literature on GWAS interpretation, quantitative trait loci analysis, epigenomic annotation, single-cell and spatial transcriptomics, multi-omics integration, artificial intelligence and machine learning, experimental validation, RNA therapeutic modality selection, delivery assessment, safety evaluation, and biomarker-informed precision medicine. Results: Genetic and multi-omics data can nominate disease-relevant lncRNAs, but no single evidence layer is sufficient to establish causality, mechanism, druggability, or clinical utility. AI can integrate heterogeneous biomedical datasets, rank candidate lncRNAs, detect regulatory patterns, and prioritize transcripts for validation. However, computational prediction should be interpreted as decision support rather than proof of therapeutic relevance. Candidate targets require disease-context expression validation, functional perturbation, mechanistic assessment, appropriate model systems, therapeutic modulation, delivery-feasibility assessment, safety evaluation, and patient-selection strategies. Conclusions: LncRNAs represent a promising but challenging therapeutic target class. A responsible translational pipeline should connect non-coding genetic evidence and multi-omics support with AI-guided prioritization, experimental validation, RNA therapeutic strategy selection, delivery assessment, safety evaluation, and clinical implementation. The framework defines qualification criteria and decision gates to reduce premature target claims during translational development.

Biomedicines doi: 10.3390/biomedicines14081720

Authors: Anamaria Ardelean Roxana Furău Florina Buleu Nicoleta Mirica Oana Todut Ion Petre Izabella Petre Tiberiu Buleu Daian-Ionel Popa Mircea Iurciuc Oana Suciu Cristian George Furău

Background: The adverse cardiometabolic outcomes that have been associated with early menopause include obesity, insulin resistance, and metabolic syndrome. Other studies have also linked metabolic dysfunction with vitamin D deficiency. However, the independent association of serum 25-hydroxyvitamin D [25(OH)D] concentrations with metabolic syndrome and type of menopause is unclear. Thus, the present study was designed to assess the association between vitamin D status and metabolic syndrome in women with early menopause compared with those with natural menopause. Methods: A total of 301 postmenopausal women were enrolled. Metabolic syndrome was assessed independently according to the NCEP-ATP III and IDF diagnostic definitions. They were classified into two groups: early menopause (EM, n = 79) and physiological menopause (PHYSM, n = 222). Demographic, anthropometric, biochemical, and lifestyle characteristics were recorded. The criteria for diagnosing metabolic syndrome were those of the National Cholesterol Education Program Adult Treatment Panel III (NCEP-ATP III) and the International Diabetes Federation (IDF). Serum 25(OH)D was also categorized as deficiency, insufficiency, or sufficiency. Multivariable logistic regression analyses were used to determine independent predictors of metabolic syndrome. Multivariable linear regression analyses were used to determine the association between serum 25(OH)D and individual metabolic parameters. Results: Women with early menopause had a higher body mass index than those with physiological menopause, whereas current smoking was more prevalent among women with physiological menopause. No significant differences were observed between the groups in the prevalence of metabolic syndrome, 25(OH)D concentrations, lipid profile, blood pressure, glycaemic indices, or other biochemical parameters. In multivariable analyses, menopausal status was not independently associated with metabolic syndrome according to either the NCEP-ATP III or IDF criteria. Body mass index was the only independent predictor of metabolic syndrome in the adjusted IDF model (OR 1.2, 95% CI 1.0–1.4; p = 0.047). Serum 25(OH)D concentrations were independently associated only with lower fasting glucose levels and were not independently associated with metabolic syndrome or its other components. Although vitamin D categories showed significant associations with metabolic syndrome in unadjusted analyses, these associations were attenuated after adjustment for demographic, anthropometric, menopausal, and lifestyle-related factors. Conclusions: Menopausal type was not independently associated with the prevalence of metabolic syndrome or vitamin D status. Although metabolic syndrome prevalence differed across vitamin D categories in unadjusted analyses, these associations were attenuated after adjustment for demographic, anthropometric, menopausal, and lifestyle-related factors. Obesity appeared to be the strongest independent predictor of metabolic syndrome within this cohort. Given the retrospective observational design, these findings should be interpreted cautiously, and prospective longitudinal studies are warranted to confirm these associations.

Biomedicines doi: 10.3390/biomedicines14081719

Authors: Noha Osama Abdellateef Mohamed Amna Fathalrahman Altayeb Mohammed Ensaf Alrsheed Sulman Mohmeed Asawer Mansour Yahia Daoud Tsabeeh Ali Mohammed Altoum Alseydhkhadiga Abdalbagi Babiker Nasir Enas Ataallah Hussien Jah Elrasoul Enaam Hussein Mohamed Ahmed Awad Elkareem Abass Mahmmoud Shagun Agarwal Ashwani Bhardwaj Lola Izzatullaevna Makhmudova Ashok Kumar Sah Ayman Husein Mohamed Alfeel

Background: Vitamin D deficiency is highly prevalent in patients with sickle cell disease (SCD) and may contribute to hematological dysregulation and increased vaso-occlusive crisis (VOC) burden. However, data from Sudan, a region with a substantial SCD burden, remain limited. This study investigated the prevalence and determinants of vitamin D deficiency and its association with hematological parameters and VOC frequency in Sudanese patients with SCD. Methods: A cross-sectional observational study was conducted among 50 patients with confirmed SCD attending a hematology outpatient clinic in Sudan. Serum 25-hydroxyvitamin D [25(OH)D] levels were measured using chemiluminescence immunoassay and categorized as deficient (<20 ng/mL), insufficient (20–29 ng/mL), or sufficient (≥30 ng/mL). Clinical and laboratory data, including hemoglobin concentration, annual VOC frequency, genotype, hydroxyurea use, hospitalization frequency, and sun exposure, were collected. Statistical analyses included chi-square tests, independent t-tests, Pearson correlation, and multivariable linear regression. Results: Vitamin D deficiency was identified in 54.0% of participants, whereas only 16.0% demonstrated sufficient vitamin D levels. Mean serum 25(OH)D concentration was 21.23 ± 7.44 ng/mL. HbSS patients exhibited significantly lower vitamin D levels compared with HbAS patients (19.89 ± 6.96 vs. 25.03 ± 7.71 ng/mL, p = 0.031). Serum 25(OH)D levels positively correlated with hemoglobin concentration (r = 0.584, p < 0.001) and inversely correlated with annual VOC frequency (r = −0.692, p < 0.001). Multivariable regression demonstrated that serum vitamin D was independently associated with hemoglobin levels (β = 0.048, p = 0.002) and annual VOC frequency (β = −0.121, p < 0.001) after adjustment for age, sex, genotype, sun exposure, and hydroxyurea use. Sun exposure was significantly associated with vitamin D status (p < 0.001). Conclusions: Vitamin D deficiency is highly prevalent among Sudanese patients with SCD and is significantly associated with anemia severity and increased VOC frequency. Given the single-center design and modest sample size, these findings should be regarded as exploratory and hypothesis-generating. They support routine vitamin D assessment and indicate that targeted supplementation strategies warrant evaluation as potential adjunctive approaches in SCD management in prospective, adequately powered studies.

Biomedicines doi: 10.3390/biomedicines14081718

Authors: Zoe Pasvanis Antony Boynes Roy C. K. Kong Elsa C. Chan Raymond C. B. Wong Jennifer Fan Gaskin

Background: Glaucoma filtration surgery (GFS) is performed to slow down disease progression in glaucoma, a leading cause of irreversible blindness worldwide. Following surgery, pathological wound healing may lead to conjunctival fibrosis and filtering failure. Myofibroblast transition of human Tenon’s fibroblast (HTF) is responsible for postoperative conjunctival scarring, with TGFβ signalling playing a key role in this process. However, the detailed molecular mechanism underlying myofibroblast transition in HTF remain understudied. Methods: In this study, we performed transcriptomic analysis to delineate the TGFβ1-induced changes in the transcriptome of HTF. HTF was isolated from three patients following GFS and treated with TGFβ1 for 5 days, followed by RNA-seq for transcriptomic profiling. Results: Our results identified 3362 differentially expressed genes (DEGs) in HTF following TGFβ1 treatment, of which 1532 were upregulated and 1820 were downregulated. Using gene ontology analysis, we identified signalling pathways associated with the pathogenesis of conjunctival fibrosis, including pathways involved in myofibroblast differentiation, TGFβ-signalling, collagen and extracellular matrix organisation, epithelial to mesenchymal transition, and cell cycle regulation. This study provided detailed characterisation of the transition from HTF to myofibroblast, and highlighted key genes that are upregulated (LDLRAD4, CDKN2B, FZD8, MYOZ1) and downregulated (SOD3, LTBP4 and RCAN2) in this myofibroblast transition process. Conclusions: Overall, this study provided insights into the transcriptional landscape of HTFs and myofibroblast differentiation, which has important implication to understanding the pathophysiology of conjunctival scarring and development of new therapeutic agents.

Biomedicines doi: 10.3390/biomedicines14081717

Authors: Wei-Luen Yen Yi-Lin Chiu Hsin-Chung Lin Hsuan-Wei Chen

Background and Objectives: In Taiwan, hepatitis B virus (HBV) or hepatitis C virus (HCV) infections are the most common causes of hepatocellular carcinoma (HCC). However, an increasing number of non-HBV and non-HCV (NBNC) patients developing HCC has been noted in recent years. The pathogenic connection between NBNC status and HCC development remains largely elusive. This study aims to explore the differences in clinical manifestations and pathological findings among HCC patients with HBV, HCV, and NBNC etiologies. Methods: This retrospective study analyzed 521 HCC patients with complete hepatic viral profiles at a single medical center in Taiwan between 2016 and 2020. Differentially expressed gene (DEG) analysis, xCell stromal cell estimation, and Gene Set Enrichment Analysis (GSEA) were employed to explore the distinct oncogenic mechanisms between the viral and NBNC groups. Results: The final cohort included 38 NBNC-HCC patients. Clinically, the NBNC-HCC patients exhibited a significantly lower FIB-4 index than the HCV-HCC patients (3.191 vs. 6.077, p = 0.0019). Furthermore, fewer NBNC-HCC patients presented with imaging-defined cirrhosis compared to HBV-HCC patients (44.4% vs. 68.1%, p = 0.0057), and a lower proportion had high Ishak scores (5–6) compared to HCV-HCC patients (31.3% vs. 75.6%, p = 0.0025). DEG analysis revealed differential expression in oncogenes (OIT3, AKR1B10) and liver fibrosis-related genes (COLEC10, CXCL14, and LINC01093). Subsequent GSEA and stromal cell analyses highlighted significant differential enrichment in hepatic stellate cells and vascular endothelial cells. Conclusions: NBNC-HCC patients present with significantly lower rates of cirrhosis and fibrosis compared to their viral counterparts. The distinct gene expression profiles and cell-type enrichment features observed between the viral and NBNC groups indicate a divergent, etiology-specific pathogenesis driving NBNC-HCC.

Biomedicines doi: 10.3390/biomedicines14081715

Authors: Jingwen Zeng Jialing Zhang James Schulz Azhar Dzulhadj B. Arafah Sora Lee Michelle Yam Yi Shen Fanfan Zhou Ting Zhang Mark C. Gillies Ling Zhu

Background: Regional heterogeneity of the macula and peripheral retinas contributes to the differences in retinal physiology, metabolic demand and susceptibility to macular diseases such as age-related macular degeneration (AMD), diabetic macular oedema (DME) and Macular telangiectasia type 2 (MacTel). Extracellular vesicles (EVs) released from the retinal pigment epithelium (RPE)-choroid are increasingly recognised as mediators of extracellular communication and correlate with the molecular biological states of their tissue of origin. However, regional variation in EV composition in human RPE-Choroid tissues remains poorly characterised. Methods: Human macular and peripheral RPE-choroid explants from non-diseased donor eyes (n = 4) were cultured ex vivo using a transwell system. EV and EV-depleted conditioned media were collected by differential ultracentrifugation, respectively. EVs were characterised by transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA). Proteomic profiling was performed using LC-MS/MS followed by multivariate, pathway, and differential expression analyses. Results: TEM and NTA confirmed the presence of vesicle-like particles in a 119–140 nm size range, indicating small EVs in both regions. Proteomic analysis demonstrated a region-associated separation between the macular and peripheral samples in both EV proteome and EV-depleted soluble secretome datasets. GO enrichment analysis revealed that macular EV proteomes were enriched for wound healing, cell–substrate adhesion, and focal adhesion-related terms with a high abundance of integrin and annexin family members, whereas peripheral EV proteomes were enriched for retinoid- and vitamin-binding terms. In the EV-depleted soluble secretome, macular-enriched proteins were associated with actin binding and extracellular matrix-related terms, while peripheral-enriched proteins were correlated with RNA localisation and nuclear compartment terms. Comparative analysis identified EV-specific, secretome-specific, and shared extracellular protein pools. Several EV-specific markers and membrane proteins such as annexins and integrins, showed a relatively high enrichment in macular EV proteomes, whereas CD63 is more abundant in peripheral EV proteomes. In contrast, EV-depleted soluble secretome contains cytokine- and ligand-associated proteins, including MIF, SPP1, and IL6, which may suggest that EVs and soluble secreted proteins represent partially distinct extracellular signalling compartments. Conclusions: Human macular and peripheral RPE-choroid explants released secretory proteins in a regional differentiated manner, supported by PLS-DA and GO enrichment analyses of both EV proteomes and EV-depleted soluble secretome. Further analysis of the EV proteomes may show that such differences were also able to be reflected in protein categories such as EV-specific markers, mitochondrial and membrane proteins. These findings provide a foundation for future investigations into the role of EV-mediated communication in macular diseases and may support the development of region-specific extracellular biomarkers and therapeutic targets.

Biomedicines doi: 10.3390/biomedicines14081716

Authors: Vincent G. Yuan

Bone remodeling is increasingly recognized as an immunologically regulated process in which inflammatory signaling governs the balance between bone formation and resorption. While extensive efforts have focused on pathways that promote osteoclastogenesis, endogenous mechanisms that restrain inflammatory bone destruction remain less well defined. Here, we propose NLRP12 as a previously underappreciated osteoimmune checkpoint that integrates innate immune regulation with skeletal homeostasis. Emerging evidence demonstrates that NLRP12 suppresses NF-κB and MAPK signaling, antagonizes NLRP3 inflammasome activation, and limits the production of osteoclastogenic cytokines, thereby constraining pathological bone resorption. Beyond its direct effects on osteoclast precursors, NLRP12 may shape the broader bone marrow immune niche through regulation of macrophages, dendritic cells, neutrophils, immunometabolic pathways, and host–microbiota interactions. We synthesize current knowledge linking NLRP12 to osteoclast differentiation, inflammatory bone diseases, and osteoimmune communication, and highlight key unanswered questions regarding its functions in osteoblasts, osteocytes, and skeletal aging. By framing NLRP12 as an important regulator of inflammatory tone within the skeletal microenvironment, we introduce an osteoimmune checkpoint paradigm that provides new conceptual insights into the pathogenesis of osteolytic disorders and identifies opportunities for therapeutic intervention.

Biomedicines doi: 10.3390/biomedicines14081714

Authors: Omar W. Maher Norhan M. El-Sayed El-Sayed E. El-Awady Naglaa F. El-Orabi Asmaa Radwan

Background and Objectives: Pulmonary fibrosis (PF) is a life-threatening respiratory disorder involving complex pathophysiological mechanisms such as inflammation, collagen deposition, and epithelial cell injury. Methotrexate (MTX), a chemotherapeutic and immunosuppressive agent, is frequently associated with pulmonary toxicity, representing a significant adverse effect with unpredictable outcomes. Thus, the current study investigates the possible protective effect of dulaglutide (DUL) against MTX-induced lung injury. Methods: Sixty male Wistar rats were allocated into six groups: group 1 served as a control group, group 2 received MTX (14 mg/kg/week, p.o) for 2 weeks, group 3 was treated with MTX treatment (14 mg/kg/week, p.o) prior to dexamethasone (DEXA) (0.5 mg/kg/week i.p), Groups 4–5 were treated concurrently with MTX (14 mg/kg/week, p.o) and DUL at doses of 0.05 and 0.1 mg/kg/week, s.c. and Group 6 received DUL (0.1 mg/kg/week, s.c.) only. Animals were sacrificed on day 15 for histopathological and biochemical assessments. Results: Our data demonstrated that MTX markedly increased some oxidative stress markers, inflammatory biomarkers and fibrotic and apoptotic indicators, alongside a significant decrease in Beclin-1 and Adenosine Monophosphate Activated Protein Kinase (AMPK) levels. DUL administration significantly ameliorated these alternations in a dose-dependent manner, with histopathological findings corroborating biochemical data through Hematoxylin & Eosin (H&E) and Mason’s Trichrome (MTC) staining. Conclusions: DUL confers significant protection against MTX-induced pulmonary fibrosis, likely through its antioxidant, anti-inflammatory, and anti-apoptotic properties, which are associated with the restoration of total AMPK and Beclin-1 levels. This highlights its therapeutic potential in preventing drug-induced lung toxicity.

Biomedicines doi: 10.3390/biomedicines14081713

Authors: Ping Zheng Aihong You Yong Fan

Background/Objectives: The NLRP3 inflammasome–Caspase-1–IL-1β pyroptotic axis participates in peripheral inflammatory responses, yet its function in peripheral–central neuroimmune crosstalk underlying endometriosis (EM)-associated pain remains unclear. This study aimed to clarify whether NLRP3-mediated pyroptosis establishes a brain–lesion neuroimmune axis connecting ectopic lesion inflammation with central neuroimmune remodeling and to explore the therapeutic mechanism of transcranial direct current stimulation (tDCS). Methods: An EM rat model was established to detect NLRP3 pathway expression in ectopic lesions and anterior cingulate cortex (ACC), together with central nervous system pathological alterations. Animals received tDCS intervention to evaluate inflammatory, neuropathological and pain behavioral changes. The closed-loop brain–lesion regulatory circuit was further interpreted. In a clinical cohort including 40 EM patients, pain and quality-of-life scores were compared between active and sham tDCS groups. Results: NLRP3, Caspase-1 and IL-1β were upregulated in ectopic lesions and ACC of EM rats, accompanied by ACC mitochondrial injury, microglial activation and thalamic demyelination. tDCS inhibited pyroptosis-related molecules, decreased systemic proinflammatory cytokines, improved central pathological lesions and relieved pain hypersensitivity. Mechanically, top-down descending pain inhibitory pathways, vagal cholinergic anti-inflammatory pathway and the HPA axis jointly mediate therapeutic effects, whereas circulating cytokines and visceral afferents transmit peripheral inflammatory signals to the brain. Clinical data demonstrated that active tDCS effectively alleviated EM-related pain and improved patients’ quality of life. Conclusions: NLRP3-mediated pyroptosis acts as a key mediator linking peripheral and central neuroimmune communication. Targeting this pathway via tDCS interrupts the inflammation–pain vicious cycle through multiple neuroregulatory pathways and remodels the central neuroimmune microenvironment in endometriosis.

Biomedicines doi: 10.3390/biomedicines14081712

Authors: Junqi Zhang Sian Xie Yongjun Wang

Colorectal cancer (CRC) is a major cause of cancer morbidity and mortality worldwide, and metabolic reprogramming is increasingly recognized as an important feature of its progression. Among these changes, fatty acid metabolism (FAM) has drawn growing attention because it supports energy supply, membrane synthesis, redox balance, and stress adaptation in tumor cells. CRC cells can increase fatty acid uptake, activate de novo synthesis, adjust fatty acid oxidation (FAO), and alter lipid droplet (LD) dynamics according to metabolic demand. These processes are strongly influenced by the hypoxic tumor microenvironment. Under hypoxic conditions, signaling pathways centered on hypoxia-inducible factors (HIFs) reshape lipid uptake, synthesis, oxidation, and storage, allowing CRC cells to maintain survival and adapt to limited oxygen and nutrient availability. Increasing evidence suggests that this metabolic shift is closely linked to invasion, metastasis, stem-like behavior, and resistance to therapy. In this review, we provide an integrated overview of the hypoxia–FAM axis in CRC. We first summarize the major steps of FAM reprogramming, then highlight how hypoxia reshapes these processes through HIF-dependent and related pathways. We also discuss FAM crosstalk with stromal and immune cells, experimental models, and metabolic heterogeneity between primary CRC and liver metastases. Finally, we discuss therapeutic strategies targeting FAM and hypoxia-associated signaling in CRC.

Biomedicines doi: 10.3390/biomedicines14081711

Authors: Lidia Boldeanu Alice Elena Ghenea Marius Bogdan Novac Virgilios Galatis Rodica Pădureanu Mohamed-Zakaria Assani Vlad Pădureanu George G. Mitroi Ancuța-Ramona Boicea Camen Mihail Virgil Boldeanu

Atopic dermatitis (AD) is a chronic inflammatory skin disease characterized by epidermal barrier dysfunction, immune dysregulation, and marked clinical heterogeneity. Growing evidence implicates the gut microbiome in AD-related pathways through microbial metabolites, intestinal barrier function, and systemic immune signaling. This narrative review synthesizes current evidence on gut microbial alterations in AD, with particular attention to short-chain fatty acids, tryptophan-derived aryl hydrocarbon receptor ligands, intestinal permeability, gut–skin microbiome interactions, and microbiota-targeted interventions. Human studies have reported associations between AD and altered abundance of selected microbial taxa, metabolite profiles, and markers of intestinal barrier dysfunction, whereas animal and in vitro studies provide complementary mechanistic evidence. However, findings remain heterogeneous across age groups, disease phenotypes, geographic populations, analytical platforms, and treatment exposures, and causality is incompletely established. Probiotic and synbiotic interventions have shown strain-specific and context-dependent effects, while postbiotics, fecal microbiota transplantation, washed microbiota transplantation, and metabolite-directed approaches remain investigational. AI-assisted multi-omics methods may improve biological stratification and hypothesis generation, but current applications are limited by small sample sizes, cohort heterogeneity, overfitting, insufficient external validation, and limited clinical implementation. Current evidence therefore supports the gut microbiome as a mechanistically plausible contributor, potential biomarker, and therapeutic target in AD while underscoring the need for longitudinal, phenotype-aware, and externally validated studies before routine clinical translation.

Biomedicines doi: 10.3390/biomedicines14081710

Authors: Wenzhe Hao Zhigang Wang Yandong Zhang Zheng Qu Dongting Chen Gang Song

Background: Prostate cancer (PCa) exhibits strong multidimensional heterogeneity. The dynamic evolution of Luminal subsets and their crosstalk with the tumor microenvironment (TME) during malignant progression remain poorly understood. Conventional single-cell transcriptomics lacks spatial context, while low-resolution spatial technologies cannot resolve single-cell and subcellular interactions. This study aimed to map a high-resolution spatial transcriptomic atlas to characterize Luminal subset heterogeneity and stromal crosstalk across progressive PCa lesions. Methods: We used high-resolution CosMx spatial molecular imaging (SMI) to profile 6 treatment-naive PCa patients and constructed a single-cell spatial transcriptomic atlas covering 385 fields of view and 154,168 high-quality cells. Unsupervised clustering, functional enrichment, cell–cell communication analysis, and flow cytometry were performed. Results: We identified 28 cell clusters, including 8 Luminal subsets (0–7) and 9 Fibroblast subsets (0–8). Pathological microregion-specific spatial distribution was observed: Luminal 0–2 dominated in BPH/PIN, while Luminal 3–7 were enriched in poorly differentiated PCa and vacuolated adenocarcinoma. These subsets were functionally linked to androgen resistance, EMT, and immune regulation. COLLAGEN, FN1, and LAMININ were identified as key pathways mediating pathological-specific tumor–stromal crosstalk. Flow cytometry verified T-cell exhaustion and immunosuppression in the TME. Conclusions: This study delineated pathological microregion-specific Luminal subset plasticity and dynamic epithelial–stromal communication across the full spectrum of PCa lesions. The uncovered molecular signatures offered mechanistic clues linking EMT, hormone resistance, and local immune status during lesion evolution. Our spatial atlas provided preliminary molecular evidence to support pathological stratification and targeted precision therapeutic strategies for PCa.

Biomedicines doi: 10.3390/biomedicines14081709

Authors: Andrei Surguchov

Type 2 diabetes mellitus (T2D) is increasingly recognized as a major risk factor for Alzheimer’s disease (AD), supporting the concept that chronic metabolic dysfunction contributes to neurodegeneration. Recent advances have identified caveolin-1 (CAV-1), the principal structural protein of caveolae, as an important regulator of insulin signaling, lipid metabolism, mitochondrial homeostasis, neurovascular integrity, and amyloid precursor protein processing. Since our previous review published in 2020, substantial evidence has demonstrated that altered CAV-1 expression and function are associated with AD-related pathology under diabetic conditions through multiple mechanisms, including endothelial dysfunction, impaired brain insulin signaling, disruption of mitochondria–endoplasmic reticulum contact sites (MERCSs), neuroinflammation, mitochondrial dysfunction, and defective autophagy. Experimental studies further show that restoring neuronal or endothelial CAV-1 expression improves insulin signaling, preserves synaptic function, attenuates amyloid pathology, and ameliorates cognitive decline in preclinical models. This review summarizes recent advances in understanding of the CAV-1-dependent mechanisms linking T2D and AD and discusses the emerging potential of CAV-1 as a biomarker and therapeutic target for diabetes-associated neurodegeneration. Collectively, current evidence identifies CAV-1 as a central molecular hub integrating metabolic, vascular, and neurodegenerative pathways and supports its further investigation as a promising therapeutic target.

Biomedicines doi: 10.3390/biomedicines14081708

Authors: Shengnan Zhou Ming Zhang Shaobei Bai Jinxiu Liu Chunyan Zhang Shuangli Mi Jian Zhang

Background: Carotid plaque rupture is a critical event in ischemic stroke, yet the potential involvement of the intraplaque microbiota across disease stages remains unclear. Methods: We performed dual-omics profiling by analyzing host transcriptomes and PathSeq-derived microbiomes from 48 human carotid RNA-seq specimens spanning early lesions (intimal thickening; n = 10), stable plaques (n = 20), and unstable plaques (n = 18). Host transcriptomes were profiled alongside intraplaque microbiomes extracted via the GATK PathSeq pipeline with rigorous in silico decontamination. We integrated differential expression analysis, microbial diversity metrics, and functional inference. Furthermore, an integrated machine learning approach (incorporating Boruta feature selection) was employed to identify exploratory cross-kingdom diagnostic biomarkers. Results: Microbial beta diversity diverged significantly across disease stages, accompanied by the progressive upregulation of 54 host genes critical for extracellular matrix remodeling and immune chemotaxis. Strikingly, despite the inherent noise and artifacts associated with low-biomass sequencing, we computationally detected the distinct enrichment of 21 bacterial taxa in unstable plaques, predominantly oral and gut mucosal pathobionts. Computationally inferred functional profiling revealed that these unstable plaque-associated microbiota were significantly linked to predicted cell death, IL-17, and HIF-1 signaling pathways and exhibited strong positive correlations with host matrix-degrading transcripts. Statistical modeling suggested associative links among specific microbial enrichment, host transcriptomic dysregulation, and plaque instability, highlighting concurrent biological cross-talk. Importantly, our integrated machine learning pipeline established a 14-feature cross-kingdom biomarker panel (10 host genes and 4 bacteria) that discriminated stable from unstable plaques (cross-validated AUC = 0.869). Conclusions: Intraplaque microbiome dynamics computationally associate with host transcriptomic alterations during carotid plaque evolution. This synergistic host–microbiome association provides a hypothesis-generating framework linking microbial dysbiosis to plaque destabilization, offering novel mechanistic insights and highlighting the exploratory cross-kingdom biomarker panel as a highly promising foundation for future experimental validation and stage-tailored clinical diagnostics.

Biomedicines doi: 10.3390/biomedicines14081707

Authors: Takayuki Yokota Masumi Iketani Toui Tsuchiya Kosuke Sakamoto Yasuhiro Nakao Tsutomu Mitsuie Eri Inoue Kota Inoue Tomoaki Kusaka Takayuki Wakabayashi Kosuke Koyano Takanori Miki Masaki Ueno Shinji Nakamura Takashi Kusaka

Background/Objectives: Hydrogen gas (H2) inhalation has shown neuroprotective effects in neonatal hypoxic–ischemic models. However, its impact on pulmonary endothelial activation and respiratory function after hypoxic–ischemic insult remains unclear. This study investigated whether H2 inhalation augments pulmonary endothelial activation or impairs pulmonary function during the acute phase after neonatal hypoxic–ischemic injury. Methods: Sixteen newborn Camborough® piglets within 24 h of birth were subjected to hypoxic–ischemic insult and randomized to an untreated group (HI, n = 8) or an H2-treated group (HI-H2, n = 8). The HI-H2 group received 2.1–2.7% H2 for 6 h. Pulmonary ICAM-1 and inducible nitric oxide synthase (iNOS) expression were assessed by immunofluorescence, lung neutrophils were quantified histologically, and pulmonary function was evaluated using arterial blood gases, oxygen index, and alveolar–arterial oxygen difference. The relationship between lung neutrophil counts and right ventricular cardiac output was also examined. Results: H2 inhalation did not increase pulmonary ICAM-1 expression or iNOS induction compared with untreated animals. Although lung neutrophil counts were significantly higher in the HI-H2 group, pulmonary gas exchange remained preserved, with no significant differences in arterial blood gases, oxygen index, or alveolar–arterial oxygen difference. Lung neutrophil counts were positively correlated with right ventricular cardiac output, suggesting that enhanced pulmonary perfusion may contribute to neutrophil redistribution rather than inflammatory recruitment. Conclusions: H2 inhalation preserved pulmonary endothelial homeostasis without impairing respiratory function during the acute phase after neonatal hypoxic–ischemic insult. The observed increase in lung neutrophils was not accompanied by evidence of endothelial activation or deterioration of gas exchange, supporting the acute pulmonary safety of H2 inhalation while suggesting that neutrophil accumulation may, at least in part, reflect hemodynamic redistribution rather than injurious inflammatory infiltration.

Biomedicines doi: 10.3390/biomedicines14081706

Authors: Gabriella Milan Chiara Compagnin Isabel Zucal Giuseppe Perale Silvia Bettini Anna Pilatone Marco De Monti Yves Harder Corrado Parodi Daniel Schmauss Vincenzo Vindigni Franco Bassetto Oliver Felthaus Dmytro Oliinyk Lukas Prantl Luca Busetto

Background: Multiple symmetric lipomatosis (MSL) is an orphan disease characterized by pathological expansion of lipomatous tissue (LT) in the subcutaneous adipose tissue (SAT) areas, causing severe deformities and compression of vital organs. To date, the only treatment options available are either lifestyle modification or surgical excision/liposuction. Since sex hormones play an important role in adipose tissue (AT) expansion, they may represent an opportunity for pharmacological treatment. Methods: LT and control SAT were collected from eight MSL patients. Both tissues and derived stromal vascular fraction (SVF) were compared in terms of aromatase (hCYP19A1) and 5-α-reductase isoform (hSRD5A1, -2 and -3) mRNA expression. Primary cultures obtained from one MSL patient were exposed to an aromatase inhibitor, 5-α-reductase inhibitor and DL-α-Lipoic Acid and assayed for cell viability, proliferation and adipogenic differentiation. Results: hCYP19A1, hSRD5A1 and hSRD5A3 were expressed in both MSL-affected and MSL-spared tissues at different levels. Cell viability and proliferation of LT-derived SVF cells were slightly increased by treatment with DL-α-Lipoic Acid and the aromatase inhibitor. Adipogenesis was slightly reduced by Finasteride and strongly reduced by Exemestane (more in LT- than in SAT- derived primary cultures). Conclusions: These preliminary results pave the way for the use of aromatase and 5-α-reductase inhibition as a potential pharmacological approach to tackling dysfunctional adipogenesis in MSL. Therefore, the reported study should be interpreted as exploratory and further experiments are recommended in a larger number of patients to validate these promising results.

Biomedicines doi: 10.3390/biomedicines14081705

Authors: Samuel Fernández-Lorenzo Cristian Marín-Pagán Lorena Ponce Juan Gambini Remus Iulian Lupu Francisco Javier Martínez-Noguera Javier Escobar

Physical performance can be understood as a continuum throughout the life course, ranging from peak athletic ability in early life to the preservation of mobility and functional independence in old age. This narrative review explores whether the biological and genetic pathways involved in athletic performance might also modulate the risk of geriatric motor dysfunctions (GMDs), a conceptual umbrella proposed here for sarcopenia, dynapenia, lower-limb weakness and the motor component of physical frailty. The available evidence suggests a convergence between performance and motor decline in mechanisms such as mitochondrial function and mitophagy, anabolic–catabolic balance, oxidative stress and low-grade chronic inflammation, neuromuscular integrity, satellite cell function, mechanotransduction, myokine-mediated signalling, and the gut–muscle axis. Although classic candidate genes such as ACTN3, ACE and PPARGC1A have been useful for formulating mechanistic hypotheses, genome-wide association studies support a highly polygenic architecture for strength, lean mass, muscle weakness and frailty. These effects are strongly modulated by the exposome, particularly by physical activity, nutrition and comorbidities. Overall, the relationship appears consistent with predominantly beneficial pleiotropy, although context-dependent effects cannot be ruled out. Genetics may influence functional reserve and decline trajectories, but exercise, particularly strength and power training, along with adequate nutrition and the management of comorbidities, remain the primary strategies for preventing or delaying sarcopenia, frailty and lower-limb weakness.

Biomedicines doi: 10.3390/biomedicines14081704

Authors: Maria V. Chatziathanasiadou

Gliomas are a broad category of primary brain tumours accounting for 80% of all malignant central nervous system (CNS) tumours [...]

Biomedicines doi: 10.3390/biomedicines14081703

Authors: Miri Pevzner Dror Sasson Chen Porat Daniel Porat Arik Dahan

Medical cannabis use is steadily increasing worldwide, including among pregnant and breastfeeding women. This Perspective provides an overview of current evidence regarding cannabis exposure during pregnancy and lactation and its implications for clinical decision-making. The available literature indicates relatively consistent evidence linking prenatal Δ9-tetrahydrocannabinol (THC) exposure with adverse perinatal outcomes, including reduced birth weight, increased risk of preterm delivery, and potential neurodevelopmental effects, although the findings remain heterogeneous and often confounded. In contrast, evidence regarding cannabis exposure during breastfeeding is sparse, and long-term infant outcomes associated with postnatal exposure through breast milk remain insufficiently characterized. Data on isolated cannabidiol (CBD) exposure during pregnancy and lactation are particularly limited, and most available evidence derives from preclinical studies or pharmacokinetic modeling rather than clinical outcome studies. Current recommendations from major health organizations uniformly advise avoiding cannabis and cannabinoid use during pregnancy and breastfeeding. This Perspective highlights the need to interpret the evidence according to cannabinoid type, exposure period, route and pattern of use, and the distinction between recreational exposure and regulated medical use. Such distinctions are particularly important because evidence is relatively stronger for prenatal THC exposure, whereas data on CBD and lactational exposure remain limited and are derived largely from indirect clinical, pharmacokinetic, and preclinical sources. Within this evidence landscape, individualized clinical interpretation may be needed when exposure has already occurred, cessation is not immediately feasible, or maternal treatment decisions remain complex, while remaining anchored in current avoidance recommendations.

Biomedicines doi: 10.3390/biomedicines14081702

Authors: Chase M. Walton Ben A. Strickland

Background/Objectives: Glioblastoma (GBM) is the most common and lethal primary malignant brain tumor in adults, with a median survival of approximately 15 months despite maximal multimodal therapy. The complement system plays a paradoxical dual role in GBM, mediating both antitumor immunity and immunosuppressive signaling within the tumor microenvironment, yet no systematic synthesis of complement-targeted therapeutic strategies exists. We aimed to comprehensively identify, appraise, and synthesize studies investigating complement-targeted therapies and complement-associated prognosis in GBM. Methods: Following PRISMA 2020 guidelines, we searched PubMed/MEDLINE and the Cochrane Library (CENTRAL) without date or language restrictions. Preclinical and clinical study designs were eligible. Risk of bias was assessed using SYRCLE, ROBINS-I, and study-type-specific checklists. Certainty of evidence was evaluated using GRADE. Statistical pooling was planned only for sufficiently comparable studies; clinical prognostic studies were synthesized narratively because they assessed non-equivalent constructs. Results: Forty-one studies were included, comprising 15 preclinical in vivo, 13 preclinical in vitro, 6 clinical observational, and 7 bioinformatics studies. Five preclinical survival studies entered a structured quantitative synthesis, but no pooled cross-target effect was calculated because their interventions, comparators, and reported summary measures were non-equivalent. Clinical prognostic studies evaluated either individual protein biomarkers or multigene immune-risk signatures and were not pooled. GRADE certainty was “Very Low” for both outcomes. C3b opsonization and the C5a/C5aR1 axis were among the most frequently studied targets. Conclusions: Complement modulation remains a promising biological hypothesis in GBM rather than evidence for clinical application, and certainty of evidence is very low. Methodological and mechanistic heterogeneity across complement targets underscore the need for standardized preclinical models and randomized clinical trials.

Biomedicines doi: 10.3390/biomedicines14081701

Authors: Anjali Kariyarath Valappil Seung-Nam Kim

Autism spectrum disorder (ASD) is a heterogeneous neurodevelopmental condition characterized by impairments in social communication, restricted and repetitive behaviors, sensory dysregulation, and frequent psychiatric comorbidities. Increasing attention has been directed toward acupuncture as a complementary neuro-modulatory intervention; however, its underlying molecular mechanisms remain incompletely understood. This review synthesizes evidence from preclinical, clinical, and molecular studies published between 2015 and 2025 to examine how acupuncture influences neurobiological pathways relevant to ASD. Current evidence indicates that acupuncture modulates multiple neurotransmitter systems, including glutamatergic, GABAergic, dopaminergic, serotonergic, and noradrenergic signaling, while also influencing neurotrophin-mediated plasticity, neuroinflammatory responses, and synaptic function. Studies conducted directly in ASD models demonstrate regulation of excitatory/inhibitory balance, monoaminergic signaling, neurotrophin pathways, and ASD-associated behavioral outcomes, whereas evidence from related neuropsychiatric conditions provides complementary mechanistic support for these pathways. Collectively, the findings suggest that acupuncture may act through coordinated modulation of interconnected neurotransmitter and neuroplasticity networks rather than a single molecular target. However, direct mechanistic evidence in ASD-specific models and clinical populations remains limited, and considerable heterogeneity exists among acupuncture protocols and outcome measures. Future studies integrating standardized stimulation paradigms with molecular, electrophysiological, neuroimaging, and behavioral assessments will be essential to validate the proposed mechanisms and clarify the translational potential of acupuncture in ASD.

Biomedicines doi: 10.3390/biomedicines14081700

Authors: Özen Toranbeki Mehmet Gagari Caymaz Atalay Elver Meryem Güvenir

Background/Objectives: Pericoronitis is a polymicrobial, biofilm-associated odontogenic infection; however, selected facultative Gram-positive bacteria may contribute to oral infectious persistence. This study compared the residual antibacterial activity of chlorhexidine (CHX), low-level laser therapy (LLLT), injectable platelet-rich fibrin (i-PRF), and amoxicillin-loaded i-PRF against selected oral and odontogenic bacterial strains. Methods: Standardized planktonic suspensions of Streptococcus mutans ATCC 25175, Staphylococcus aureus ATCC 25923, and Enterococcus faecalis ATCC 29212 were prepared at 0.5 McFarland turbidity. Five conditions were evaluated: 0.2% CHX, i-PRF, amoxicillin-loaded i-PRF, 976 nm diode LLLT, and 0.9% sodium chloride as the negative control. Twelve independent tubes were used for each treatment and species, yielding 180 experimental units. After a residual 24-h exposure/incubation period, samples were serially diluted, plated on 5% sheep blood agar, and quantified as CFU/mL. Data were analyzed using non-parametric tests. Results: Significant treatment-dependent differences were observed for all species (p < 0.001). CHX showed the most consistent antibacterial activity, with median residual counts of 550 CFU/mL for S. mutans, 1 CFU/mL for S. aureus, and 5.5 CFU/mL for E. faecalis. i-PRF alone was comparable to the negative control. Amoxicillin-loaded i-PRF markedly reduced S. aureus counts, whereas S. mutans and E. faecalis remained at high residual levels. LLLT showed partial, variable inhibition mainly against S. aureus. Conclusions: CHX demonstrated the most consistent antibacterial effect in this planktonic residual 24-h in vitro model. i-PRF alone showed no measurable antibacterial activity, while amoxicillin-loaded i-PRF and LLLT exhibited limited, species-dependent effects.

Biomedicines doi: 10.3390/biomedicines14081699

Authors: Bing Song Zhangrong Zhan Hua Gao Hui Che Yuefeng Hao You Li Dan Hu

Objectives: Frozen shoulder (FS) is characterized by chronic inflammation and fibrosis of the joint capsule, yet the causal roles of immune cell subtypes remain unclear. This study aimed to identify immune cell traits causally associated with FS and evaluate the potential impact of glucocorticoids (GCs) on these immune signatures. Methods: A two-sample Mendelian randomization (MR) analysis was performed using genome-wide association study data for 731 immune cell traits and FS. RNA sequencing (RNA-seq) of FS and control capsular tissues was performed to validate immune cell-related signatures and immune infiltration patterns. MR analysis was further used to investigate the relationship between GCs, FS, and immune cell traits. Results: MR identified seven immune cell traits as potential risk factors and 17 as potential protective factors in FS. RNA-seq confirmed the up-regulation of five of these traits, which positively correlated with the inflammatory, fibrotic and angiogenic features of FS pathology. Notably, GCs showed no causal association with FS but were negatively associated with a protective immune cell trait. Conclusions: Our findings reveal a complex immune cell landscape underlying FS, characterized by both pathogenic and protective immune traits. The integration of MR and RNA-seq analyses highlights potential therapeutic targets and suggests that the immunological consequences of GC therapy warrant further investigation.

Biomedicines doi: 10.3390/biomedicines14081698

Authors: Marta Mallardo Furqan Memon Ludovica D’Auria Raffaella Pagliaro Aurora Daniele Ersilia Nigro

Background: Colorectal cancer (CRC) remains one of the most prevalent and lethal malignancies worldwide. Adiponectin, a hormone secreted by adipose tissue, has been increasingly recognized for its pleiotropic effects in several malignancies. In particular, an inverse association between circulating adiponectin levels and CRC incidence supports a potential protective role for adiponectin and its receptors (AdipoR1, AdipoR2). The aim of this study was to investigate the effects of adiponectin on Caco-2 cells, a human CRC cell line, by examining intestinal epithelial differentiation using AdipoRon (AR), a synthetic adiponectin receptor agonist. Methods: The effects of AR on Caco-2 cells were assessed by evaluating dome formation and the expression of key molecular markers involved in intestinal epithelial differentiation, including KLF-4, DPPIV, SI, and KRT20, at both transcriptional levels using qRT-PCR and at protein levels using immunofluorescence and Western blot analysis. In addition, mitochondrial reactive oxygen species (ROS) production was assessed using MitoSOX™ Red. Results: Our findings showed that AR administration was associated with a dose-dependent increase in AdipoR1 and AdipoR2 expression as well as with dome formation in Caco-2 cells. Furthermore, AR administration reduced Ki-67 expression with no changes in cell cycle and an increase in the differentiation markers DPPIV and SI, while KRT20 remained unchanged. Finally, AR decreased mitochondrial ROS levels during differentiation. Conclusions: Our findings provide new evidence that AR contributes to intestinal epithelial differentiation and may represent a potential therapy for colorectal cancer. AdipoRon may contribute to the regulation of intestinal epithelial differentiation and may contribute to restoring epithelial homeostasis in tumor cells. Further research is needed to clarify the underlying mechanisms and assess the translational relevance of adiponectin-based therapies.

Biomedicines doi: 10.3390/biomedicines14081697

Authors: Sarah Bergren Hannah Shelby Julian Wier Edward M. Schwarz Denis Evseenko Jay R. Lieberman

Osteoarthritis (OA) is a significant clinical problem that places a substantial burden on both patients and the healthcare system. Characterized by progressive cartilage degeneration, synovial inflammation, and subchondral bone remodeling, OA is a rapidly growing and increasingly studied disease affecting millions around the world. Growing evidence has expanded on the traditional view of OA as a mechanical “wear-and-tear” disease, highlighting that disease progression is driven not only by mechanical stress but also by chronic dysregulation of the osteoimmune environment. Activation of innate and adaptive immune pathways, macrophage M1 polarization, and dysregulated cytokine signaling all contribute to progressive joint degeneration. While current therapeutics often focus on managing symptoms or restoring joint mechanics, interventions often overlook the role of osteoimmunology in disease progression. This review summarizes the biological and bioengineering strategies emerging to address OA. These platforms include bioceramics, metal-based scaffolds, hydrogels, nanoparticles, and microsphere systems. Furthermore, small molecules, cell-based therapies, and gene-modified systems have also been shown to modulate the inflammatory microenvironment, enhance regulatory immune responses, and restore cartilage homeostasis. Together, these approaches represent the evolving research landscape, shifting away from just symptom alleviation and towards targeted disease-modifying therapies, including osteoimmunomodulation. However, significant barriers to clinical translation remain, such as limited large animal studies and species immune system differences, which need to be addressed for the development of clinically applicable interventions.

Biomedicines doi: 10.3390/biomedicines14081696

Authors: Federica Li Pomi Maria Clara Gama de Souza Silva Giuseppe Murdaca Francesco Borgia Adele Bottaro Sebastiano Gangemi Alessandro Allegra

Malondialdehyde (MDA) is one of the main end-products of lipid peroxidation (LPO) and represents a widely investigated marker of oxidative stress (OS) and oxidative tissue damage. Beyond its role as a measurable byproduct of polyunsaturated fatty acid peroxidation, MDA can interact with proteins and nucleic acids, generating adducts that may contribute to mutagenic, genotoxic, and cytotoxic events involved in carcinogenesis and tumor progression. This narrative review summarizes current evidence on the role of MDA in cancers, including breast, lung, head and neck, colorectal, cervical, and cutaneous tumors. Across these cancer types, increased circulating or tissue MDA levels have frequently been associated with enhanced LPO, impaired antioxidant defenses, tumor burden, advanced disease stage, aggressive histopathological features, treatment-related oxidative injury, and, in selected studies, poorer clinical outcomes. MDA-derived DNA adducts may further reflect oxidative DNA damage and provide mechanistic insight into the relationship between chronic redox imbalance, inflammation, and malignant transformation. However, MDA remains a non-specific biomarker influenced by age, smoking, diet, metabolic disorders, systemic inflammation, comorbidities, treatment exposure, and analytical methodology. Current evidence therefore supports MDA as a biologically relevant indicator of oxidative damage rather than a validated stand-alone diagnostic, prognostic, or therapeutic biomarker. Larger prospective studies using standardized and specific analytical methods are needed to clarify its clinical utility and to integrate MDA within broader redox biomarker panels.

Biomedicines doi: 10.3390/biomedicines14081695

Authors: Yuting Zhu Xun Hu Zhuo Shi Yuan Liang Dengfeng Li Peiqing Ma Dong Yan Jianwei Liang Qian Wang

Aim: To establish and validate a quantitative transporter-mediated imaging framework based on gadoxetic acid-enhanced T1 mapping for assessing liver functional reserve (LFR) and to investigate the physiological significance of the relative change in longitudinal relaxation rate (ΔR1%) as a quantitative imaging biomarker. Methods: Female C57BL/6J mice (6–8 weeks old) representing five experimental liver conditions (control, transporter-deficient Slco1b2/Slco1a5 double-knockout, carbon tetrachloride-induced fibrosis, methionine–choline-deficient diet-induced steatohepatitis, and alcohol-associated fatty liver disease; n = 6 per group) underwent serial Gd-EOB-DTPA-enhanced T1 mapping. Quantitative ΔR1% was calculated to characterize hepatobiliary enhancement kinetics. Liver functional reserve was independently evaluated using multispectral optoacoustic tomography of indocyanine green (ICG) pharmacokinetics and serum ICG retention assays, with histopathological and hepatocellular transporter analyses performed for mechanistic validation. Longitudinal data were analyzed using restricted maximum likelihood (REML)-based mixed-effects models. Intergroup comparisons were performed using one-way ANOVA or Kruskal–Wallis tests with appropriate post hoc analyses, and associations between imaging and functional parameters were evaluated using Spearman rank correlation analysis. A two-sided p < 0.05 was considered statistically significant. Results: Five experimental liver models exhibited distinct transporter-dependent hepatobiliary enhancement patterns. The transporter-deficient knockout mice showed minimal enhancement, whereas fibrosis and steatotic liver injury models demonstrated intermediate but clearly distinguishable functional profiles. Longitudinal mixed-effects analysis identified significant effects of time, experimental group, and time-by-group interaction on ΔR1% dynamics (all p < 0.0001). Although MRI-derived ΔR1% parameters were not significantly correlated with regional optoacoustic ICG kinetics, ΔR1% area under the curve showed a strong inverse correlation with serum ICG retention at 600 s (r = −0.729, p < 0.0001), indicating that MRI-derived ΔR1% and ICG-based measurements provide complementary rather than interchangeable assessments of liver function. Histological and molecular analyses further demonstrated marked heterogeneity in fibrosis, steatosis, and hepatobiliary transporter expression across models, whereas transporter abundance alone did not consistently predict imaging-derived functional performance. Conclusions: Quantitative Gd-EOB-DTPA-enhanced T1 mapping provides a transporter-mediated imaging framework for evaluating hepatic functional reserve across mechanistically distinct liver injury models. As a normalized quantitative imaging biomarker, ΔR1% captures the integrated functional consequences of hepatobiliary transport dysfunction and complements established liver function tests. These findings support the translational potential of quantitative T1 mapping as a standardized, noninvasive approach for assessing liver functional reserve.

Biomedicines doi: 10.3390/biomedicines14081694

Authors: Monika Rudkowska Jarosław Mołdoch Olga Wronikowska-Denysiuk Monika Agacka-Mołdoch Anna Pradiuch Karolina Wojtunik-Kulesza

Background/Objectives: Voltage-gated sodium channels (VGSCs) are among the most important molecular targets in epilepsy therapy. Unlike classical antiseizure medications (ASMs), newer sodium channel modulators selectively affect slow inactivation or persistent sodium currents, potentially improving seizure control while preserving physiological neuronal activity. This review summarizes the pharmacology, mechanisms of action, clinical efficacy, and therapeutic potential of eslicarbazepine acetate, lacosamide, and cenobamate. Methods: A narrative review of published clinical trials, meta-analyses, and real-world evidence was conducted. The analysis focused on sodium channel modulation, pharmacokinetic properties, efficacy in monotherapy and adjunctive therapy, and safety profiles in focal epilepsy. Main findings of the review: Eslicarbazepine acetate and lacosamide primarily enhance slow inactivation of VGSCs, suppressing pathological repetitive neuronal firing with limited effects on normal neuronal signaling. Both agents demonstrated efficacy in monotherapy and add-on therapy, with favorable pharmacokinetic properties and a relatively low potential for drug–drug interactions. Cenobamate represents a novel therapeutic approach through preferential inhibition of persistent sodium currents combined with positive allosteric modulation of GABAA receptors. Clinical trials and real-world studies demonstrated high responder and seizure freedom rates, particularly in patients with drug-resistant focal epilepsy. The most common adverse effects across these agents included dizziness, somnolence, fatigue, and gastrointestinal symptoms, while notable safety concerns included hyponatremia with eslicarbazepine acetate and drug interactions or dose-dependent adverse effects with cenobamate. Conclusions: Recent advances in sodium channel modulation have expanded therapeutic options for focal epilepsy and support the development of more selective, mechanism-based ASM therapies. Eslicarbazepine acetate, lacosamide, and cenobamate demonstrate favorable efficacy and tolerability profiles and may improve seizure control in patients with drug-resistant epilepsy.

Biomedicines doi: 10.3390/biomedicines14081691

Authors: Marina Khodanovich Mikhail Svetlik Daria Kamaeva Anna Usova Tatyana Anan’ina Irina Wasserlauf Maria Shadrina Valentina Pashkevich Marina Moshkina Victoria Obukhovskaya Nadezhda Kataeva Anastasia Levina Valentina Alifirova Marina Titova Irina Zhukova Svetlana Vasilieva Evgeny Schastnyy Yana A. Tumentceva Vasily Yarnykh

Background/Objectives: Normal-appearing white (WM) and gray (GM) matter outside focal demyelinating lesions (WMH) has been found impaired in many diseases and normal aging. The present study aimed to compare global GM and WM alterations in patients with multiple sclerosis (MS), Parkinson’s disease, long COVID (LC) and normal aging. Methods: The study population comprised 196 participants including patients with MS (n = 42), PD (n = 16), LC (n = 75), and healthy volunteers (n = 63). All participants underwent MR scanning using the fast macromolecular fraction (MPF) mapping protocol and routine clinical sequences. MPF in global normal-appearing WM, GM, and mixed WM-GM was measured after exclusion of segmented focal lesions. To eliminate the influence of gender, age, and brain atrophy on the results, the corresponding covariates were included in the analysis. Results: Significant WMH volume increase and MPF decrease in global WM, WM-GM, and GM were found both in normal aging (75–85 years) and MS, PD, and LC patients (except for WM and WMH volume in LC patients). The greatest MPF decrease was observed in MS patients. The youngest (18–24 years) controls had a significant MPF decrease in global WM, WM-GM, and GM compared to middle-aged (35–44 years) participants. Among LC patients, the greatest global MPF reduction was found in patients with depression and insomnia as COVID-19 complications. In healthy controls, the peak age of myelination was estimated as 42.2 years for WM, 46.5 years for WM-GM, and 45.4 years for GM. Significant correlations were found between MPF and EDSS in MS patients (r = −0.71 in GM, r = −0.58 in WM-GM, r = −0.44 in WM) and stage of PD in PD patients (r = −0.58 in GM, r = −0.55 in WM-GM, r = −0.52 in WM). Conclusions: MPF mapping showed high sensitivity to age-related and disease-related differences in brain myelination, particularly in GM. Our results confirm the feasibility of using MPF mapping in large-scale clinical studies.

Biomedicines doi: 10.3390/biomedicines14081693

Authors: Ghazala Muteeb Rayan A. Siraj

Antimicrobial resistance (AMR) in respiratory infections represents a major global health challenge, compounded by biological barriers that limit the effectiveness of conventional antibiotics, including mucus hypersecretion, biofilm formation, and intracellular pathogen persistence. Nanotechnology has emerged as a promising platform for addressing these limitations through advanced drug-delivery strategies. This narrative review provides an integrated overview of nanocarrier systems—including lipid-based (e.g., liposomes, solid lipid nanoparticles), polymeric (e.g., PLGA, chitosan), and inorganic nanoparticles (e.g., silver, gold, zinc oxide)—with emphasis on their pharmaceutical design parameters for pulmonary delivery. Key mechanisms by which nanotechnology enhances antimicrobial efficacy include targeted and controlled drug delivery, improved penetration of mucus and biofilms via surface engineering, synergistic combination therapies, and intrinsic antimicrobial activity through mechanisms such as reactive oxygen species generation. Preclinical studies targeting major respiratory pathogens, including Pseudomonas aeruginosa, Mycobacterium tuberculosis, Streptococcus pneumoniae, and methicillin-resistant Staphylococcus aureus, demonstrate enhanced biofilm disruption, intracellular drug delivery, and reductions in bacterial burden. However, important translational challenges remain, including long-term safety, manufacturing scalability, regulatory complexity, and the potential for microbial adaptation. Future directions focus on stimuli-responsive systems, inhalable formulations, and biomimetic platforms to improve targeting and therapeutic precision. Collectively, nanotechnology represents a delivery-oriented strategy with the potential to enhance existing antimicrobial therapies and support the development of more effective interventions against resistant respiratory infections.

Biomedicines doi: 10.3390/biomedicines14081692

Authors: Yanyan Chen Jingni Zhu Jinghang Qian Sheng Li Liu Yang

The myocyte enhancer factor 2 (MEF2) transcription factor family plays crucial roles in differentiation, lineage specification, stress responses, and tissue homeostasis. Recent investigations have shown that the dysregulation of MEF2A, MEF2B, MEF2C, and MEF2D is associated with tumorigenesis, tumor progression, and adverse clinicopathological features in several cancers. MEF2B has a particularly important role in B-cell malignancies, where recurrent mutations deregulate BCL6 and promote lymphoma progression. MEF2A, MEF2C, and MEF2D also regulate malignant phenotypes, including proliferation, migration, invasion, apoptosis, drug resistance, angiogenesis, inflammation, and immune evasion, by the mechanism of regulating cell-cycle regulators, apoptosis-related genes, EMT-related genes, and other transcriptional programs. This review summarizes the mechanisms by which MEF2 family members contribute to tumor initiation and progression, with added emphasis on MEF2B mutations and MEF2D fusions. We also discuss clinical associations with overall survival and recurrence in solid tumors and hematologic malignancies. Because MEF2 proteins are transcription factors with broad physiological functions, we evaluate therapeutic strategies: RNA interference, genetic perturbation, small molecules that alter MEF2-dependent transcription, and PROTAC or oligonucleotide-PROTAC. These platforms remain promising but require MEF2-specific validation, tumor-selective delivery, and careful toxicity assessment.

Biomedicines doi: 10.3390/biomedicines14081690

Authors: Yafei Kang Peiling Li Xue Zhang Xinyi Dong Suying Yuan Xu Wen Zhixuan Yang Xiaoxue Yang Xiaoqing Zhang Hu Zhao Yonghong Tian Donghui Huang

Objectives: This study aimed to investigate the mechanisms underlying impaired sperm motility in high-fat diet (HFD)-induced obesity. Methods: Six-week-old male C57BL/6J mice were used to establish an HFD-induced obesity model. Sperm motility and male reproductive function were then evaluated. Macrophage polarization was evaluated by flow cytometry, and key differentially expressed genes were identified by RNA-seq analysis of epididymal adipose tissue, the epididymis, and testes. Finally, pyruvate carboxylase (PC) and the PC inhibitor erianin (Eri) were used to examine the role of adipocyte-derived PC in regulating macrophage polarization. Results: HFD-induced obese mice exhibited abnormal testicular and epididymal morphology, with reduced sperm motility and viability. Flow cytometry revealed that macrophages were polarized toward the M1 phenotype in epididymal adipose tissue and the epididymis in the HFD group. RNA-seq analysis demonstrated that PC was the most significantly upregulated gene in epididymal adipose tissue. Furthermore, PC expression increased during adipocyte maturation in vitro. Mature adipocytes promoted macrophage polarization toward the M1 phenotype, while Eri attenuated this effect. In addition, supernatants from LPS-induced M1 macrophages contained elevated levels of inflammatory cytokines and significantly reduced sperm motility and viability. Conclusions: HFD-induced obesity impaired sperm quality and male reproductive function. The underlying mechanism may involve increased PC expression, which drives epididymal macrophages toward M1 polarization and triggers local chronic inflammation, ultimately compromising sperm motility and viability.

Biomedicines doi: 10.3390/biomedicines14081689

Authors: Mengru Cai Yu Qiu Mingkai Yao Jiahui Kong Xiang Li Qian Zhang Yiman Jia Zicheng Zhu Yukun Zhao Dong Bai Yuqin Yang

Plant-derived exosome-like nanoparticles (PELNs) are lipid bilayer nanostructures containing endogenous lipids, proteins, nucleic acids, and phytochemicals, which have attracted increasing interest as bioinspired carriers for cancer therapy. This review evaluates how plant source, isolation, purification procedures, vesicle composition, cargo-loading strategy, and administration route shape the quality and tumor-delivery performance of PELNs. The available evidence indicates that plant source and processing are major determinants of particle size, purity, surface charge, cargo profile, and biological activity. Ultracentrifugation remains widely used but is limited by contaminant co-isolation and poor scalability, whereas density-gradient purification and size-exclusion chromatography improve purity, and ultrafiltration and tangential flow filtration offer greater potential for large-scale manufacturing. Passive incubation generally preserves vesicle integrity and is most suitable for hydrophobic small molecules, whereas electroporation, sonication, and extrusion can increase cargo loading but may cause aggregation, membrane remodeling, or loss of endogenous components. Preclinical studies suggest that PELNs can exert intrinsic antitumor effects, modulate the tumor microenvironment, improve chemotherapeutic delivery, and help overcome drug resistance. However, evidence for in vivo tumor-targeting remains less robust than evidence for cellular uptake, and direct comparisons with established nanocarriers remain scarce. Clinical translation will require standardized nomenclature and characterization, reproducible manufacturing, quantitative loading and release assays, route-specific biodistribution studies, and repeated-dose safety evaluation. These findings provide a framework for the rational development of PELNs as reproducible tumor-oriented nanocarriers.

Biomedicines doi: 10.3390/biomedicines14081688

Authors: Daniel Cernach Ayres Ruan Pimenta Gabriel Arantes dos Santos Patrícia Candido Milena Antunes Nelson Gaspar Dip Junior Giovanni S. Marchini Fábio C. Torricelli Fábio C. Vicentini Alexandre Danilovic Carlos A. Batagello Kátia R. M. Leite William C. Nahas Sabrina T. Reis Eduardo Mazzucchi

Introduction: Cystinuria is an inherited disorder characterized by impaired proximal tubular reabsorption of cystine, resulting in recurrent cystine stone formation. Mutations in the SLC3A1 and SLC7A9 genes are the main genetic causes of the disease, while variants in PBX1 have also been associated with cystinuria in specific populations. Despite advances in understanding its molecular basis, cystinuria remains clinically heterogeneous, making disease progression and prognosis difficult to predict. This study aimed to investigate circulating microRNAs (miRNAs) as potential molecular biomarkers, characterize their expression profile in patients with cystinuria, and evaluate their association with clinical indicators of disease severity. Methods: The study group included 16 patients with a clinically established diagnosis of cystinuria, while the control group consisted of 11 healthy individuals without a history of nephrolithiasis. Candidate miRNAs were selected based on their predicted regulatory interactions with genes implicated in cystinuria pathogenesis. Eight microRNAs (miR-29a-3p, miR-29b-3p, miR-29c-3p, miR-1207-3p, miR-10a-3p, miR-3658, miR-141-3p, and miR-200a-3p) were analyzed by quantitative real-time PCR (qPCR). Results: Among the eight miRNAs evaluated, miR-10a, miR-141, miR-200a, and miR-3658 were significantly downregulated in patients with cystinuria compared with healthy controls. Higher expression levels of miR-10a, miR-141, and miR-200a were significantly associated with a greater number of previous surgical procedures (p < 0.05). Additionally, increased miR-10a expression was associated with impaired renal function, defined as serum creatinine levels above 1.2 mg/dL. Conclusions: miR-10a, miR-141, miR-200a, and miR-3658 were significantly downregulated in patients with cystinuria compared with healthy controls. Within the cystinuria cohort, higher expression of miR-10a was associated with impaired renal function, whereas increased expression of miR-10a, miR-141, and miR-200a correlated with a greater number of surgical interventions, suggesting a potential relationship with disease severity. Further studies in larger, genetically characterized cohorts are warranted to clarify the biological role and clinical utility of these miRNAs as biomarkers of cystinuria.

Biomedicines doi: 10.3390/biomedicines14081687

Authors: Grace Clouse Samantha Penman Aidan Powell Faisal Sadar Nihamul Ehan Isaiah T. Taylor Ayanna Varma Michael Hadjiargyrou David E. Komatsu Panayotis K. Thanos

Introduction: Osteoporosis is a debilitating bone disease that affects a large segment of the population. Currently, pharmacological treatment options for osteoporosis, including antiresorptive drugs and those that prevent bone loss, are limited. Multiple studies have shown that the endocannabinoid system and cannabis may play a significant role in bone remodeling. As both the use of cannabis and high-quality research on its effects continue to grow, novel medicinal uses of selected components, such as tetrahydrocannabinol (THC), are being investigated. The present study examined the potential effect of chronic THC inhalation during early adulthood on an ovariectomized rodent model of post-menopausal osteoporosis. Materials and Methods: Young (26-week-old) adult female rats received daily either THC or air inhalation for 8 weeks. The rats then underwent an ovariectomy (OVX) or sham surgery (Sx) when they were 34 weeks old. They were then euthanized 4 weeks post-OVX, and skeletal tissues were collected for analysis. Results: MicroCT analysis of femora showed that THC + OVX rats showed a significant decrease (45%) in trabecular bone volume compared to Air + Sham Sx rats. As expected, the Air + OVX rats also showed a 17% reduction in trabecular bone volume. Discussion: Overall, early adulthood THC exposure increased susceptibility to osteoporosis following ovariectomy-induced menopause. This suggests that THC exerts long-lasting effects on bone health, which may exacerbate post-menopausal bone loss.

Biomedicines doi: 10.3390/biomedicines14081686

Authors: Jianhua Mu Yitian Wang Han Liu Xuanhong He Zhuangzhuang Li Minxun Lu Fan Tang Yi Luo Yong Zhou Li Min Chongqi Tu

Background: Although the tumor coagulome interacts with the tumor immune microenvironment (TME) in solid tumors, its role in osteosarcoma (OS) remains uncharacterized. This study aimed to delineate this transcriptomic interplay and identify potential prognostic targets. Methods: This study was performed with bulk RNA sequencing (RNA-seq), single-cell RNA sequencing (scRNA-seq), and clinical phenotype data. Bioinformatic approaches were employed at the transcriptomic level to investigate the impact of the tumor coagulome on the TME and prognosis in OS. We validated the above findings using immunohistochemistry and immunofluorescence. Results: The activity of a coagulation-related transcriptional signature was found to correlate with the degree of malignancy in OS. Its activity score demonstrated predictive value for OS prognosis, with a maximum area under the curve (AUC) of 0.802. scRNA-seq analysis indicated that inflammatory cancer-associated fibroblasts (iCAFs) and APOE+ macrophages were predominantly enriched in the high coagulation score subgroup. Our data further suggest that iCAFs may facilitate the M2 polarization of APOE+ macrophages via the C3–C3AR1 axis, potentially contributing to poorer clinical outcomes in patients with OS. Conclusions: These findings imply that within a high coagulation-related transcriptional score group, the interaction between iCAFs and APOE+ macrophages, likely mediated by the C3–C3AR1 axis, could facilitate OS progression. Consequently, the C3–C3AR1 signaling pathway might represent a promising target for future therapeutic strategies and coagulome monitoring in OS.

Biomedicines doi: 10.3390/biomedicines14081685

Authors: Carlos Fernandez-Pereira Alfredo E. Rodriguez

Background: Coronary stenting remains the cornerstone of interventional cardiology. Drug-eluting stents (DES) have reduced restenosis compared to bare-metal stents (BMS), but their high cost and need for prolonged dual antiplatelet therapy (DAPT) limit accessibility in many regions. Recent evidence has revisited the potential role of systemic pharmacologic adjuncts—oral sirolimus (rapamycin), prednisone and colchicine—as cost-effective therapies to mitigate restenosis and adverse events following BMS implantation. Objective: This review critically evaluates the clinical and mechanistic evidence supporting the use of oral immunosuppressive or anti-inflammatory drugs following BMS implantation, with emphasis on their applicability in both resource-limited and general cardiology settings. Three randomized clinical trials comparing this strategy against DES are analyzed and discussed in this review. Conclusions: Oral sirolimus, prednisone, and colchicine demonstrate promising anti-inflammatory and antiproliferative effects that translate into clinical outcomes comparable to DES in a highly select population. Their systemic administration following BMS implantation may provide a feasible, cost-effective alternative where DES use is restricted by cost or clinical contraindications. Larger-scale, long-term trials are warranted to confirm safety, optimize dosing, and identify ideal patient populations for this “pharmacologic stent hybrid” strategy.

Biomedicines doi: 10.3390/biomedicines14081684

Authors: Danial Qasim Butt Maaz Anwer Memon Masitah Hayati Harun Shazana Hilda Shamsuddin Nur Asyilla Binti Che Jalil Saidi Jaafar Teffanie Arputheraj Basaruddin Ahmad

Introduction: DNA aptamers are single-stranded nucleic acids capable of selectively binding target proteins and have emerged as potential alternatives to antibodies in molecular diagnostics. This study aimed to develop and characterize in silico designed DNA aptamers targeting Interferon alpha-inducible protein 6 (IFI6) and Radical S-adenosyl-L-methionine domain-containing protein 2 (RSAD2) for oral squamous cell carcinoma (OSCC). Methods: Genomic transfer RNA sequences from Homo sapiens, Mus musculus, and Escherichia coli were computationally truncated and optimized to generate DNA aptamer candidates ranging from 35–50 mers. Secondary and tertiary structures were generated using Mfold and RNAComposer, followed by molecular docking with AutoDock Vina and molecular dynamics simulations in GROMACS to evaluate docking interactions and structural behavior comparatively. Selected DNA aptamer candidates were synthesized and evaluated by aptahistochemistry (AHC) to qualitatively explore preliminary tissue reactivity in formalin-fixed paraffin-embedded OSCC tissues under optimized conditions. Results: Molecular docking demonstrated favorable comparative docking scores ranging from −15.6 to −18.7 kcal/mol. RMSD analysis demonstrated that selected IFI6 and RSAD2 DNA aptamer–protein complexes reached a plateau with relatively small fluctuations following maximum RMSD values. In contrast, RMSF analysis identified increased flexibility predominantly within loop regions. Cross-reactivity analysis explored preliminary target selectivity, with no interactions observed within the selected target-binding regions. During AHC optimization, the 50-IFI6 and 45-RSAD2 DNA aptamer candidates exhibited cytoplasmic brown granular staining in >50% of OSCC tumor cells under the optimized experimental conditions. Conclusions: The findings support the applicability of an integrated in silico workflow for generating structurally optimized DNA aptamer candidates and provide preliminary proof-of-concept for their exploratory histopathological application in OSCC.

Biomedicines doi: 10.3390/biomedicines14081683

Authors: Ceren Pamukcu Ahmet Emin Atik

Background/Objectives: Glycation, a non-enzymatic post-translational modification, can increase structural heterogeneity in therapeutic monoclonal antibodies (mAbs) and is considered a critical quality attribute in biosimilar (BS) development. Despite extensive studies on glycation in individual mAbs, comparative reports on originator (OR) and BS mAbs under forced glycation conditions remain limited. This study aimed to comparatively evaluate the glycation profiles of one OR and three BS anti-tumor necrosis factor-alpha (anti TNF-α) mAb products using integrated mass spectrometry-based methods. Methods: Forced glycation was induced by incubating mAbs with 200 mM D-glucose at 37 °C for 7 days. Intact mass analysis and peptide mapping were used to assess glycation extent and site distribution, respectively. Results: Intact mass analysis revealed a consistent mass increase of approximately 486 Da across all major glycoform species for each mAb product, indicating predominant formation of the tri-glycated mAb population under the applied stress conditions. The overall glycation levels were comparable at the intact level, ranging from 67% to 73% among the OR and BS mAb products. Peptide mapping identified nine glycated lysine (K)-containing peptides, among which three major glycation hotspots (LC:V5 K145/K149, LC:V7 K183, and HC:V7 K250/K252) showed elevated occupancies (~10–16%). These sites collectively accounted for the dominant intact level mass shift. The remaining glycated peptides exhibited only minimal modification levels (<2%). Despite their distinct manufacturing processes, all tested mAb products showed nearly identical site-specific glycation profiles. Conclusions: Forced glycation susceptibility in the studied mAbs was driven by a limited set of structurally preferred K hotspots, leading to highly comparable glycation profiles across OR and BS mAb products. The combined intact mass and peptide mapping strategy provides a robust analytical platform for comparative glycation assessment in BS characterization.

Biomedicines doi: 10.3390/biomedicines14081682

Authors: Kanwarjot Singh Dervens Michaud Tal Parness Odinaka Mgbeke Henry Okodaso Kwami Jones Bawo Teddy Ikolo Shellon Thomas Felicia Ikolo

Sickle cell disease (SCD) is a monogenic hemoglobinopathy in which hemoglobin S (HbS) polymerizes on deoxygenation, leading to sickling of red blood cells (RBCs) and driving the acute vaso-occlusive crisis (VOC). With curative (transformative) therapies inaccessible to most patients in sub-Saharan Africa and the Caribbean, where burden is highest, optimizing acute VOC care is a priority; yet oxygen is given reactively, after hypoxemia is documented, not at symptom onset when polymerization is most interruptible. We conducted a structured narrative review integrating HbS polymerization kinetics, clinical studies of oxygen-based therapies, trial protocols, and home-care implementation evidence (PubMed, Scopus, Google Scholar; 1974–2026). When RBCs release oxygen, HbS does not solidify at once: a brief pause, the nucleation delay phase (tD), precedes polymerization and sickling. Because tD falls steeply as deoxygenated hemoglobin rises (inversely, to its ~30th–50th power), a small early gain in oxygen saturation (~5–10%) lengthens it many-fold, opening a short window, roughly 30 min from onset, during which oxygen may abort a crisis. The clinical message is that timing, not dose, is decisive: oxygen at the first symptoms may prevent a crisis that the same oxygen, given later, cannot. Standard, high-flow, hyperbaric, and inhaled nitric-oxide modalities help in selected hospital settings, and the DREPADOM model shows community delivery before hospital arrival is feasible and safe. The central argument is precision in timing, not quantity: starting oxygen at symptom onset could turn a low-cost, universally available treatment into a crisis-aborting one for patients beyond the reach of curative therapy. We advance this as a mechanistically grounded but clinically untested hypothesis; regional implementation trials in the Caribbean and sub-Saharan Africa are urgently warranted.

Biomedicines doi: 10.3390/biomedicines14081681

Authors: Jeongmi Lee Seung Hyeun Lee

The therapeutic paradigms for both non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC) have entered an unprecedented era of precision oncology, a transformative evolution fundamentally driven by the seamless integration of translational science [...]

Biomedicines doi: 10.3390/biomedicines14081680

Authors: Rajashree Patnaik Yajnavalka Banerjee

Background: Factor Xa (FXa) is increasingly recognized as a non-hemostatic signaling protease that amplifies inflammatory responses through protease-activated receptor-2 (PAR-2), but its contribution to osteoarthritis-relevant chondrocyte signaling remains incompletely defined. A central unresolved question is whether an endogenous ligand of FXa can selectively dampen its receptor-mediated signaling output. Objectives: We investigated whether palmitoylcarnitine (PalCar), an endogenous long-chain acylcarnitine previously shown to bind FXa, modulates FXa-driven inflammatory and catabolic responses in bone marrow mesenchymal stem cell (BMSC)-derived chondrocytes, and the extent to which that output is hard-wired through PAR-2. Methods: BMSC-derived chondrocytes, validated histologically and by COL2A1 expression, were challenged with FXa (1 U/mL, 24 h) alone or together with PalCar (0.5 and 1.0 µg/mL). Cytokine, catabolic and signaling responses were assessed by RT-qPCR, Western blotting and ELISA; surface RANK and RANKL by flow cytometry; and mitochondrial membrane potential by Rhodamine 123. Receptor dependence was tested by siRNA-mediated PAR-2 silencing, and data were analyzed by one-way ANOVA with Dunnett’s post hoc test. Results: FXa stimulation induced PAR-2 expression, increased TNF-α, IL-1β, and MCP-1 production, enhanced ERK1/2 activation, upregulated the catabolic mediators SOX4 and ADAMTS5, promoted surface RANK/RANKL expression, and impaired mitochondrial membrane potential. PalCar at 0.5 and 1.0 µg/mL significantly attenuated these responses at both transcript and protein levels and restored mitochondrial membrane potential under FXa-induced stress. Mechanistically, siRNA-mediated PAR-2 silencing substantially blunted, but did not completely quench, the inflammatory response to FXa, identifying PAR-2 as the dominant, though not exclusive, signaling conduit in this system. Conclusions: By combining pharmacological challenge with receptor knockdown, our data define PalCar as an endogenous, PAR-2-directed suppressor of the FXa–PAR-2–ERK inflammatory axis in chondrocytes. Given emerging evidence linking osteoarthritis with increased cerebrovascular risk, PalCar-sensitive FXa signaling may represent a mechanistically relevant intersection between joint inflammation and broader thrombo-inflammatory disease biology, providing a mechanistic basis for evaluating FXa-responsive lipid pathways in osteoarthritis and related thrombo-inflammatory states.

Biomedicines doi: 10.3390/biomedicines14081679

Authors: Michele Gottardi Federico De Marchi Giulia Ciotti Marco Basso Vittoria Raimondi Vincenzo Ciminale Giorgia Simonetti Martina Ghetti Rosa Di Liddo Roberta De Marchi Islam Ab Abouzeid Alessandra Sperotto

Acute myeloid leukemia (AML) develops within a bone marrow environment that influences leukemic stem cell behavior, residual disease, and response to therapy. This review examines evidence that the marrow microenvironment is not only a site of leukemic growth, but can actively shape AML initiation, maintenance, and treatment resistance. Clinical observations such as donor cell leukemia after allogeneic transplantation, together with experimental models in which stromal or osteolineage abnormalities induce myeloid disease, suggest that altered niches may contribute to leukemogenesis in selected settings. In established AML, vascular and endosteal compartments provide adhesive, chemokine, inflammatory, and metabolic signals that promote leukemic-cell retention, quiescence, survival, and chemotherapy tolerance. AML cells also remodel the surrounding marrow, suppressing normal hematopoiesis and generating stromal, endothelial, osteoblastic, adipocytic, and immune-cell programs that favor leukemic persistence. These interactions are especially relevant to drug resistance, including resistance to venetoclax-based therapy, where cytokine-mediated changes in apoptotic dependence, fatty-acid metabolism, mitochondrial adaptation, and stromal support may all contribute. Several therapeutic approaches have attempted to disrupt niche-mediated protection, including targeting CXCL12/CXCR4 signaling, adhesion pathways, inflammatory circuits, Hedgehog signaling, and metabolic dependencies. Although early-phase studies have shown activity in some AML subsets, randomized evidence remains limited and results have been inconsistent. We discuss how a better understanding of microenvironmental biology may help define when niche-directed therapy is most likely to complement conventional and molecularly targeted AML treatment.

Biomedicines doi: 10.3390/biomedicines14081678

Authors: Stelian Adrian Ritiu Sonia Elena Popovici Marius Papurica Dorel Sandesc Adelina Baloi Daiana Toma Norbert Wellmann Petru Bucuras Claudiu Rafael Barsac Ovidiu Bedreag

Hemorrhagic shock is a leading cause of preventable death following multiple trauma, driven by a cascade of interacting cellular and molecular disturbances that extend well beyond simple volume loss. Acute blood loss initiates tissue hypoperfusion and cellular hypoxia, setting in motion the lethal triad of hypothermia, acidosis, and coagulopathy through several converging pathways: complement activation with excessive C3a and C5a production; neutrophil-mediated tissue injury; NADPH-oxidase-driven reactive oxygen species (ROS) overproduction that overwhelms superoxide dismutase defenses; mitochondrial respiratory chain impairment; dysregulation of the pro-inflammatory cytokine network; and endothelial apoptosis with degradation of the endothelial glycocalyx and disruption of interendothelial junctions, with consequent vascular hyperpermeability. These mechanisms provide the biological rationale for the resuscitation strategy. Each class of fluid acts on these pathways in a distinct way: crystalloids modulate acid–base homeostasis, chloride-mediated renal vasoconstriction, and coagulation factor activity; colloids influence oncotic pressure, endothelial integrity, and microvascular perfusion; and blood products, particularly plasma and whole blood, actively modulate mitochondrial metabolism, endothelial permeability, and pro-apoptotic signaling beyond their volume-expanding role. Translating this biology to the bedside requires a multimodal monitoring framework that converts molecular endpoints into real-time therapeutic targets, integrating lactate and base excess as markers of cellular oxygen debt, dynamic preload indices such as pulse pressure and stroke volume variation, advanced hemodynamic platforms, point-of-care ultrasonography, viscoelastic coagulation testing, and near-infrared spectroscopy of tissue oxygenation. This review synthesizes the biological basis of hemorrhagic shock and its translation into an individualized, goal-directed resuscitation strategy for the critically ill polytrauma patient.

Biomedicines doi: 10.3390/biomedicines14081677

Authors: Jiahua Zhou An Phuc Ta Catherine Yang Ahmed El Shamy

Background/Objectives: Substance use behaviors share a complex, overlapping polygenic architecture, yet translating genome-wide association study (GWAS) findings into actionable biological mechanisms remains challenging. This study aimed to characterize the genetic architecture of five substance use traits (alcohol consumption, alcohol dependence, nicotine use, illicit drug use, and behavioral disinhibition) and identify shared and distinct gene expression signatures within the neural circuits governing addiction. Methods: We reanalyzed 7188 individuals from the Minnesota Center for Twin and Family Research (MCTFR) cohort utilizing longitudinal composite phenotypes spanning five substance-use domains and general behavioral disinhibition. Post-QC, 6874 individuals were retained for downstream analysis. Following genomic imputation and linear mixed model GWAS (GEMMA), we utilized the SNipar framework to partition polygenic risk scores (PRS) into direct and indirect genetic effects, investigating intergenerational shifts in genetic penetrance and effects of assortative mating. Finally, we integrated our summary statistics with brain tissue reference panels to perform a transcriptome-wide association study (TWAS) modeling genetically regulated gene expression within neural circuits relevant to addiction. Results: Partitioning of polygenic risk revealed that while surface-level parental DNA correlations were modest (r = 0.08), underlying latent genetic correlations approached unity (Rδ ≈ 0.99), indicating that addiction risk clustering in families is driven by intense assortive mating and concentrated biological inheritance. Multi-phenotype TWAS identified several significant gene–phenotype associations—notably ADAM32 and SLC9A3, which demonstrated pleiotropic effects across multiple substance use categories. Crucially, these significant TWAS signals were enriched in striatal structures (caudate, putamen, substantia nigra) and frontal cortical regions. Conclusions: Our findings support a model of shared genetic liability across diverse substance use behaviors, mediated by specific gene expression patterns in the mesolimbic dopamine system and frontal cortex. By integrating multi-phenotype GWAS and TWAS, this study highlights pleiotropic candidate genes and provides critical insights into the tissue-specific neurobiological pathways underlying addiction vulnerability.

Biomedicines doi: 10.3390/biomedicines14081676

Authors: Georgiana-Anca Vulpoi Tudor-Ștefan Rotaru Cătălina Elena Bistriceanu Lenuța Bîrsanu Codrina-Madalina Palade Dan Iulian Cuciureanu

Background: Musculoskeletal and peripheral nerve disorders have been reported among occupational computer users. However, the relationship between the duration of occupational computer use and ulnar neuropathy at the elbow (UNE) remains uncertain. This exploratory observational study investigated whether the duration of occupational computer use was associated with electrodiagnostically confirmed UNE and with ulnar nerve conduction parameters in a selected symptomatic population. Methods: Patients aged 23–68 years who were referred for nerve conduction studies (NCS) because of symptoms suggestive of upper-limb entrapment neuropathy were consecutively evaluated over a 1.5-year period. Among the 62 patients included, 22 met the neurophysiological diagnostic criteria for UNE, whereas 40 did not. The associations between occupational computer-use duration, bilateral ulnar sensory nerve action potential (SNAP) amplitudes, across-elbow motor nerve conduction velocities (NCV), and the presence of ulnar neuropathy were assessed using univariable and multivariable analyses. Results: Longer occupational computer-use duration showed modest unadjusted associations with lower bilateral ulnar SNAP amplitudes but was not associated with across-elbow motor NCV. In multivariable analysis, age and sex were significant independent predictors of SNAP amplitude, whereas computer-use duration was not (B = −0.086, β = −0.048, p = 0.756). Exposure duration was also not independently associated with bilateral changes in across-elbow motor NCV or with electrodiagnostically confirmed UNE. Conclusions: The findings do not support an independent association between the number of years of occupational computer use and ulnar nerve electrodiagnostic abnormalities beyond the effects of age and sex. However, small associations cannot be excluded because of the limited sample size, the indirect assessment of biomechanical exposure, and the inclusion of a selected symptomatic referral population.

Biomedicines doi: 10.3390/biomedicines14081675

Authors: Lorenzo Carriera Pier-Valerio Mari Roberto Lipsi Simone Ielo Eugenio De Corso Stefano Baglioni Alberto Ricci Luca Richeldi

The therapeutic goals related to chronic airway diseases are evolving from short-term symptom control toward sustained suppression of disease activity and prevention of future risk. In severe asthma, this shift has been captured by the concept of clinical remission, generally defined by absence of exacerbations, no need for oral corticosteroids, symptom control, and stable or improved lung function. In chronic obstructive pulmonary disease (COPD), the analogous concept has more often been described as disease stability. Although remission in asthma and stability in COPD have developed within different biological and clinical frameworks, they may reflect disease-specific expressions of the same therapeutic ambition. Recent studies support COPD stability as a measurable and clinically meaningful state, associated with reduced exacerbation risk and mortality. Evidence from optimized inhaled triple therapy, particularly with fluticasone furoate/umeclidinium/vilanterol, indicates that multidimensional stability can be achieved and maintained in a proportion of patients, while real-world studies reinforce its applicability beyond randomized trials. The emergence of biologic therapies for selected patients with eosinophilic or type 2 COPD further strengthens the rationale for considering stability as an ambitious treatment target. In this narrative review, informed by a structured literature search, we discuss the conceptual relationship between asthma remission and COPD stability, and summarize the evidence supporting disease stability as an attainable and prognostically relevant outcome. In addition, we propose a pragmatic multidimensional definition based on symptom stability, absence of moderate or severe exacerbations, no systemic corticosteroid use, and maintained lung function over 12 months. COPD stability should not be viewed as a weaker goal than asthma remission, but rather as the most appropriate COPD-specific expression of sustained low disease activity.

Biomedicines doi: 10.3390/biomedicines14081674

Authors: Thai Hau Koo Rishi Chowdhary Kirti Arora Kelly Chun Lynn Lai Andee Dzulkarnaen Zakaria

Colorectal cancer (CRC) kills approximately 850,000 people annually and is the second leading cause of cancer mortality worldwide. After curative-intent surgical resection, recurrence rates of 15–20% in stage II and 30–40% in stage III diseases highlight the fundamental limitations of pathological staging as the sole basis for adjuvant treatment decision-making. Circulating tumor DNA (ctDNA), which comprises tumor-derived cell-free DNA fragments shed into the bloodstream by dying cancer cells, offers a direct and dynamic readout of molecular residual disease (MRD) in the postoperative period. Because ctDNA clears within hours of macroscopically complete surgery, any persistently detectable postoperative signal reflects viable microscopic disease rather than surgical contamination. Over the past decade, evidence has matured rapidly from proof-of-concept observational studies to completed randomized clinical trials, culminating in the recent publication of DYNAMIC-III and ALTAIR in 2025–2026. The DYNAMIC trial established that ctDNA-guided management reduced adjuvant chemotherapy use from 27.9% to 15.3% in stage II colon cancer while maintaining an equivalent five-year recurrence-free survival (88% vs. 87%; difference 1.1%, 95% CI 5.8% to 8.0%). The GALAXY study confirmed ctDNA as the most powerful prognostic biomarker in resected CRC, with a disease-free survival hazard ratio of 11.99. Critically, DYNAMIC-III demonstrated that escalating chemotherapy intensity in ctDNA-positive stage III patients did not improve recurrence-free survival (HR 1.11, p = 0.6), and the phase III ALTAIR trial showed that trifluridine/tipiracil failed to significantly improve disease-free survival at the point of molecular recurrence (HR 0.79, p = 0.107). This review critically synthesizes the current evidence for ctDNA-guided treatment decisions in resected CRC across three therapeutic strategies: de-escalation in ctDNA-negative patients, escalation in ctDNA-positive patients, and post-adjuvant therapy at the point of molecular recurrence. Future progress requires biomarker-matched escalation strategies, adaptive trial designs, and formal validation of ctDNA clearance as a surrogate endpoint for predicting patient outcomes.

Biomedicines doi: 10.3390/biomedicines14081673

Authors: Abdurrahim Tekin Umut Çelik Akın Öztürk Salih Dere Engin Can Evren Sönmez Lokman Ayhan Suna Dilbaz Nuri Serdar Baş Serdar Çevik

Aim: The relationship between lumbar intervertebral disc degeneration and paraspinal muscle degeneration remains controversial. Although previous studies have reported associations between disc degeneration and muscle atrophy and fatty infiltration, the effect of age on this relationship has not been sufficiently clarified. This study aimed to investigate the associations that cumulative and L4–L5 level-specific disc degeneration have with paraspinal muscle morphology, and to determine whether these associations persisted after adjustment for age. Materials and Methods: This retrospective cross-sectional study included 84 patients who underwent lumbar magnetic resonance imaging (MRI) for low back pain. Disc degeneration was evaluated at all lumbar levels according to the Pfirrmann classification, and the Total Pfirrmann Score was calculated. Relative functional cross-sectional area (rFCSA) and fatty infiltration (FI) measurements of the multifidus, erector spinae, and psoas muscles at the L4–L5 level were performed using ImageJ software. The relationships between variables were examined using Spearman correlation analysis. Multiple linear regression analysis was performed to investigate independent predictors. Age-controlled partial correlation analyses were also performed to evaluate the potential confounding effect of age. Results: Of the 84 patients included in the study, 48 (57.1%) were female and 36 (42.9%) were male, with a mean age of 42.3 ± 8.4 years. The mean Total Pfirrmann Score was 14.3 ± 3.8. A moderate positive correlation was found between age and the Total Pfirrmann Score (rho = 0.519; p < 0.001). Significant positive correlations were observed between the Total Pfirrmann Score and multifidus fatty infiltration (rho = 0.316; p = 0.003) and erector spinae fatty infiltration (rho = 0.300; p = 0.006). A weak negative correlation was detected between psoas rFCSA and the Total Pfirrmann Score (rho = −0.247; p = 0.023). Multifidus and erector spinae rFCSA were not significantly associated with the Total Pfirrmann Score (p = 0.094 and p = 0.095, respectively). At the L4–L5 level, higher Pfirrmann grade was associated with lower multifidus rFCSA (rho = −0.233; p = 0.033) and lower psoas rFCSA (rho = −0.319; p = 0.003), whereas the inverse association with erector spinae rFCSA was borderline (rho = −0.215; p = 0.050). Multiple linear regression analysis showed that age was the only statistically significant predictor of the Total Pfirrmann Score in the multivariable model (p < 0.001). After adjustment for age, the associations of multifidus FI (r = 0.193; p = 0.079) and erector spinae FI (r = 0.194; p = 0.078) with the Total Pfirrmann Score did not reach statistical significance. In the age-adjusted L4–L5 analysis, only the inverse association between Pfirrmann grade and psoas rFCSA remained statistically significant (r = −0.232; p = 0.034). Conclusions: Greater cumulative lumbar disc degeneration was associated with increased fatty infiltration of the multifidus and erector spinae muscles and lower psoas rFCSA, whereas multifidus and erector spinae rFCSA were not associated with the Total Pfirrmann Score. At L4–L5, greater disc degeneration was associated with lower multifidus and psoas rFCSA, while the association with erector spinae rFCSA was borderline. After adjustment for age, the associations of multifidus and erector spinae fatty infiltration with cumulative disc degeneration were attenuated and did not reach statistical significance; however, a weak independent association cannot be excluded.

Biomedicines doi: 10.3390/biomedicines14081672

Authors: Sara Palma Gullì Rocco Morena Francesca Serapide Alessandro Russo

Background: Colonization with multidrug-resistant (MDR) Gram-negative bacteria is common in critically ill patients, yet the meaning of a positive surveillance culture is not always clear at the bedside. For some patients, colonization remains asymptomatic; for others, it may represent the first step toward an invasive infection that can develop rapidly. Objectives: This review examines the transition from colonization to infection in intensive care unit (ICU) patients carrying MDR Pseudomonas aeruginosa, carbapenem-resistant or extended-spectrum β-lactamase-producing Enterobacterales, and carbapenem-resistant Acinetobacter baumannii (CRAB). Discussion: Across these pathogens, colonization should not be viewed as an isolated microbiological finding. Its clinical relevance depends on the organism involved, site and persistence of carriage, extent of colonization, prior antibiotic exposure, severity of illness, invasive devices, impaired host defences, and the epidemiological pressure within the ICU. Intestinal carriage of resistant Enterobacterales is particularly relevant because it may act as an endogenous reservoir for subsequent invasive infection. In patients colonized with CRAB, respiratory and multisite carriage appear to identify those at greatest risk of pneumonia and poor outcomes. For Pseudomonas aeruginosa, the relationship is less uniform but becomes more relevant in the presence of respiratory vulnerability, ICU exposure, and previous broad-spectrum antibiotic use. Conclusions: Colonization results should therefore support risk stratification, infection-prevention measures, and informed empirical treatment when infection is suspected, rather than automatically leading to antibiotic therapy.

Biomedicines doi: 10.3390/biomedicines14081670

Authors: Clara F. López-León Julia Sala-Jarque José Antonio del Río Jordi Soriano

Background: Tauopathies are neurodegenerative disorders characterized by the abnormal hyperphosphorylation and aggregation of the microtubule-associated protein tau, leading to disrupted neuronal connectivity and progressive brain dysfunction. Despite their clinical relevance, most in vitro models have focused primarily on molecular and cellular aspects, with limited emphasis on alterations in network dynamics and functional connectivity. Methods: We developed an in vitro tauopathy model based on mouse primary neuronal cultures, enabling the investigation of network-level alterations under controlled conditions. We compared three experimental groups: untreated control cultures, cultures exposed to extracellular wild-type tau, and cultures treated with pathological tau (pTau) isolated from the sarkosyl-insoluble fraction of P301S (+/−) transgenic mice. To increase susceptibility to tau-induced pathology, all conditions were additionally transduced with adeno-associated viral vectors encoding human P301L tau. To quantify for damage, spontaneous neuronal activity was monitored throughout network maturation—from day in vitro (DIV) 7 to 16—using fluorescence calcium imaging, and multiple metrics describing network dynamics and functional organization were compared at DIV 12. Results: We observed that exposure to pTau did not induce overt cytotoxicity or major disruptions in global network dynamics, although a mild increase in network bursting activity was observed. Longitudinal analysis of network maturation further revealed largely similar developmental trajectories across experimental groups, with only subtle and persistent differences in bursting-related activity in pTau-treated cultures. Conclusions: We propose that the early developmental stage of the cultures, together with the intrinsic bursting and ongoing synaptic plasticity of primary neuronal networks, masks subtle pathological effects. This may limit the sensitivity of two-dimensional in vitro systems to detect network-level dysfunction, suggesting that more mature or structurally complex models, such as brain organoids, may be required to reveal robust functional deficits.

Biomedicines doi: 10.3390/biomedicines14081671

Authors: Serav Koç Şule Gül Aydın Hasan Can Toyganözü Emel Lüftüoğlu Ali Can Koç Cenk Parlatan Cevdet Adıgüzel Emre Destegül

Background: This study aimed to evaluate the clinical utility of shear wave elastography (SWE) in assessing abscess wall stiffness and its potential contribution to predicting the need for surgical intervention in patients with tubo-ovarian abscess (TOA). Materials and methods: This prospective cross-sectional study included 34 women diagnosed with TOA who underwent SWE evaluation upon admission. Demographic characteristics, laboratory parameters, abscess size, abscess wall elasticity, and abscess content elasticity were recorded. These patients were categorized as either medically managed or surgically treated. Group comparisons, receiver operating characteristic (ROC) curve analysis, and multivariable binary logistic regression were performed to identify independent predictors of surgical intervention. Results: There were no significant differences between the operated and non-operated groups regarding age, body mass index, and laboratory parameters (p > 0.05). The median length of hospital stay was significantly longer in the operated group (15 vs. 12 days, p = 0.017). Abscess diameter was significantly larger in the operated group (6.75 ± 1.17 cm vs. 4.65 ± 1.20 cm; p = 0.001). Likewise, the median abscess wall SWE value was significantly higher in the operated group (3.12 vs. 1.47 kPa; p = 0.027). The optimal cut-off value for abscess wall SWE was 2.04 kPa (AUC: 0.721, 95% CI: 0.551–0.892, sensitivity: 64.7%, specificity: 70.6%). Abscess size demonstrated a higher predictive performance (AUC: 0.905, 95% CI: 0.796–1.000). However, in the multivariable logistic regression analysis, only abscess size remained an independent predictor of surgical intervention (OR: 4.14, 95% CI: 1.50–11.39, p = 0.006), whereas abscess wall SWE and baseline C-reactive protein were not independently associated with treatment failure. Conclusions: Increased abscess wall stiffness measured by SWE was associated with the need for surgical intervention in patients with TOA. Although SWE was not an independent predictor after adjustment for abscess size and baseline C-reactive protein, it may serve as a complementary imaging biomarker when interpreted together with established clinical, laboratory, and conventional ultrasonographic findings.

Biomedicines doi: 10.3390/biomedicines14081669

Authors: Francesco Paolo Lo Muzio Lorenzo Fassina Jens Ötvös Katharina Wierling Sebastian Baer Leonhard Berboth Andreas Kind Alessandro Faragli Alessio Alogna

Background: Heart failure with preserved ejection fraction (HFpEF) is a heterogeneous syndrome requiring invasive and/or exercise stress testing for diagnosis, in contrast with the homogeneous nature of HF with reduced EF (HFrEF). We previously demonstrated that acoustic cardiography (AC), combined with advanced signal analysis, can non-invasively estimate left ventricular (LV) functional indices in an experimental swine model of HFrEF. Objectives: To evaluate whether AC-derived features capture stress-dependent LV functional changes, supporting phenotypic differentiation between porcine models of HFpEF, HFrEF, and Control conditions. Methods: Synchronized invasive LV-pressure and non-invasive ECG, pulse oximetry, and AC signals were collected from 12 anesthetized, closed-chest Göttingen minipigs (Control, n = 3; HFrEF, n = 5; HFpEF, n = 4). Through signal analysis, we derived time and frequency features from the non-invasive signals to predict, using our AI model, the invasively measured LV functional indices. Atrial pacing was performed up to 160 bpm as a controlled heart-rate stress paradigm. Two AI modeling strategies were employed: a standard 80/20 train/test ratio, and a leave-one-animal-out (i.e., 1 animal per health status) to assess generalizability. Results: Standard blind testing achieved >95% accuracy in phenotype classification with <3% relative error for predicted LV indices. The leave-one-animal-out classification performance remained robust (79–95% accuracy), supporting translational potential despite inter-animal variability. Notably, HFpEF animals exhibited greater variability in AC-features across increasing heart rates compared to Control and HFrEF. Conclusions: AC-based modelling offers a rapid, non-invasive approach for assessing LV function. Our methodology may complement existing diagnostic tools, particularly for conditions like HFpEF, but warrants further validation in clinical populations.

Biomedicines doi: 10.3390/biomedicines14081668

Authors: Da Mi Kim Zheng Shen Quan Pan Zeyu Liu Wanbao Yang Natividad R. Fuentes Robert S. Chapkin Gus A. Wright Bhimanagouda Patil Shaodong Guo Yuxiang Sun

Background: Endotoxemia is a severe inflammatory condition that is characterized by acute immune responses and oxidative stress; endotoxemia can further develop into a cytokine storm and sepsis leading to severe organ damage. Our recent studies revealed that the growth hormone secretagogue receptor (GHSR) regulates macrophage polarization in obesity- and aging-associated chronic inflammation. However, its role in acute inflammation during endotoxemia remains unclear. Methods: We subjected myeloid-specific Ghsr knockout mice (LysM-Cre;Ghsrf/f) to lipopolysaccharide (LPS)-induced endotoxemia in vivo and treated bone marrow-derived macrophages (BMDMs) with LPS in vitro. Subsequently, mouse survival rate and inflammatory signatures in the blood, peritoneal cavity, liver, and BMDM were assessed. In the ex vivo study, conditioned medium (CM) from BMDMs was applied to primary hepatocytes to assess how BMDM-derived CM influences hepatocyte inflammatory responses. Results: Myeloid-specific Ghsr knockout mice exhibited a significantly improved survival rate following LPS-induced endotoxemia, accompanied by reduced systemic inflammation, evident in the blood, peritoneal macrophages, and liver. In addition, Ghsr deficiency suppressed LPS-induced caspase-1 activation and pro-inflammatory cytokine secretion in macrophages. Consistent with these results, conditioned media from Ghsr-deficient BMDMs attenuated the inflammatory responses of primary hepatocytes. Mechanistically, LPS increased GHSR expression in BMDMs, and Ghsr-deficient BMDMs activated mitochondrial respiration and suppressed production of mitochondrial reactive oxygen species (ROS), resulting in downregulation of inflammatory activation of macrophages following LPS exposure. Conclusions: These data demonstrate that macrophage GHSR promotes systemic and tissue inflammation during endotoxemia by regulating mitochondria-associated macrophage polarization. The findings suggest that macrophage GHSR may represent a promising immunomodulatory target for acute inflammatory states, including endotoxemia and sepsis.

Biomedicines doi: 10.3390/biomedicines14081667

Authors: Shiva Hooshmandi Nicholas S. Freestone Francesca I. F. Arrigoni

Purpose: Hypertension in women is a dynamic, hormone sensitive condition shaped by cumulative physiological changes across the life course. This review summarises current evidence relating vascular ageing, hormonal regulation, metabolic dysfunction, and reproductive history to blood pressure regulation in women. Materials and Methods: A narrative review of the literature was conducted using PubMed, Scopus and Google Scholar. Clinical, epidemiological, and mechanistic studies were synthesised to evaluate factors influencing hypertension in women. Reports in which menopausal status was not defined, or previous reproductive milestones were not documented, were excluded or interpreted with caution. Results: Evidence suggests that menopause, vascular ageing, metabolic dysfunction, androgen to oestrogen balance, and reproductive history interact to influence endothelial function, neurohormonal regulation, renal sodium handling, and vascular resistance. Ageing-related mechanisms, including cellular senescence, chronic low-grade inflammation, and genetic susceptibility, may contribute to increased cardiovascular risk. Hypertensive disorders of pregnancy identify women who are at higher risk of developing cardiovascular disease later in life and provide an opportunity for earlier risk assessment and prevention. Emerging therapies, including GLP-1 receptor agonists and SGLT2 inhibitors, may offer additional options for improving cardiovascular risk management, although their role in sex-specific prevention remains an evolving area of research. Conclusions: Hypertension in women is best understood within a life-course framework. Incorporating reproductive history, menopausal status, metabolic health, and emerging risk markers may improve cardiovascular risk assessment and support earlier intervention.

Biomedicines doi: 10.3390/biomedicines14081666

Authors: Toshinori Hirai Kan Katayama

Thromboembolism is a life-threatening complication of nephrotic syndrome, which is treated with oral anticoagulants (warfarin and direct oral anticoagulants [DOACs]) that possess a variety of pharmacokinetic characteristics. This narrative review focused on drug–drug interactions between oral anticoagulants and calcineurin inhibitors (CNIs) (e.g., tacrolimus, cyclosporine, and voclosporin) for the management of thromboembolism in nephrotic syndrome. While warfarin has less potential for interacting with CNIs, DOACs carry a risk of drug–drug interactions with CNIs owing to the inhibition of drug-metabolizing enzymes and transporters, including cytochrome P450 3A4 (CYP3A4) and P-glycoprotein. Specifically, a significant increase in DOAC exposure was observed when DOACs were co-administered with cyclosporine, a more potent P-glycoprotein inhibitor than tacrolimus. Limited evidence suggests that more pronounced drug interactions occur in specific cases: (1) apixaban and CNIs in patients with renal impairment, (2) edoxaban combined with cyclosporine in patients with renal impairment, and (3) a combination of rivaroxaban with dual inhibitors of P-glycoprotein and CYP3A4 (e.g., cyclosporine and fluconazole). From a pharmacological viewpoint, CNI-induced nephrotoxicity and hypertension may increase the risk of critical bleeding in patients receiving DOACs. In addition, the variation factors of pharmacokinetic parameters, such as renal function, pharmacogenomic profiles, and pathophysiological changes directly caused by nephrotic syndrome, may vary between patients, which could potentially augment the magnitude of drug–drug interactions between oral anticoagulants and CNIs. Further studies are required to understand the clinical significance of drug interactions in patients with nephrotic syndrome.

Biomedicines doi: 10.3390/biomedicines14081665

Authors: Han Zhang Dongfeng Wei Xuejie Han Wei Wu Xingfang Liu Manman Xu

Background: Global aging has led to a rise in neurodegenerative diseases. Neuroinflammation constitutes a core pathological mechanism in these disorders. The Kongsheng Zhenzhong Pill (KSZZP), a modern preparation derived from a classical Chinese herbal formula, are reputed for their sedative and cognition-enhancing effects. However, the molecular mechanism underlying its modulation of microglia-mediated neuroinflammation remains unclear. The present study was designed to assess the intervention effects of KSZZP on LPS-induced neuroinflammation in BV-2 microglial cells and to preliminarily explore the potential molecular mechanisms involved. Methods: An in vitro neuroinflammation model was established in LPS-induced BV-2 microglial cells. The chemical components of KSZZP were identified using UPLC-Q-Exactive HFX technology. The pharmacological effects of KSZZP were evaluated by assessing cell activation, inflammatory response, oxidative stress, and apoptosis. Molecular docking and Western blotting were used to explore the specific mechanism of its action on the cGAS-STING pathway. Results: Chemical analysis identified 67 components in KSZZP, primarily flavonoids, prenyl lipids, and isoflavones. KSZZP treatment dose-dependently inhibited LPS-induced BV-2 microglial activation and significantly reduced pro-inflammatory factor release. Furthermore, it alleviated oxidative stress, mitigated mitochondrial ultrastructural damage, and inhibited apoptosis induced by LPS. Molecular docking revealed that key active components of KSZZP exhibit strong binding potential to cGAS and STING proteins. Western blotting further confirmed that KSZZP dose-dependently suppressed the expression of key cGAS-STING pathway proteins (cGAS, STING) and downstream proteins associated with M1 polarization (iNOS, TNF-α, COX-2). Conclusions: This study indicates that KSZZP alleviates LPS-induced microglial activation, neuroinflammation, oxidative stress, mitochondrial damage, and apoptosis, and these effects may involve the modulation of the cGAS-STING signaling pathway. Collectively, these findings provide a preliminary experimental basis for understanding the anti-neuroinflammatory mechanism of KSZZP and support its potential application in the prevention and treatment of neurodegenerative diseases.

Biomedicines doi: 10.3390/biomedicines14081664

Authors: Jesse Chieh-Szu Yang Yu-Hung Tian En-Rung Chiang Yu-Ping Su

Background: High tibial osteotomy (HTO) is commonly performed to manage medial compartment knee osteoarthritis by correcting mechanical alignment; however, the role of adjunctive regenerative therapies remains uncertain. Methods: This retrospective study compared leukocyte-rich platelet-rich plasma (LR-PRP) with leukocyte-poor PRP (LP-PRP) in patients undergoing HTO. Forty patients were allocated into three groups: HTO alone (n = 10), HTO with LR-PRP (n = 20), and HTO with LP-PRP (n = 10). Clinical outcomes were assessed preoperatively and at 12 months using the Visual Analog Scale, Oxford Knee Score, and Western Ontario and McMaster Universities Osteoarthritis Index. Cartilage repair appearance was evaluated through second-look arthroscopy using the ICRS grading and Koshino staging systems. Multivariable analysis of covariance (ANCOVA), adjusting for baseline imbalances, was employed to evaluate postoperative outcomes. Results: All groups demonstrated significant improvements in pain and function (p < 0.05), with no significant differences among groups. However, Group B exhibited a greater shift toward lower ICRS grades than Group A (p < 0.05), whereas no significant difference was found between Groups C and A. Arthroscopic findings revealed more complete defect coverage and improved structural integrity in the LR-PRP group. Conclusions: These findings demonstrate a clear discrepancy exists between clinical and structural outcomes; while HTO drives substantial and comparable short-term functional improvements across all cohorts, adjunctive LR-PRP is positively associated with a significantly enhanced arthroscopic cartilage repair appearance compared to LP-PRP or HTO alone. Further prospective studies are needed to validate these findings and elucidate the underlying biological mechanisms.

Biomedicines doi: 10.3390/biomedicines14081663

Authors: Crawford Currie Tor Åge Myklebust Christian Bjerknes Bomi Framroze

Background: Healthy ageing depends on the maintenance of several interconnected biological processes including adequate oxygen-carrying capacity, control of systemic inflammation, and preservation of lean body mass. ProGo® is a salmon protein hydrolysate (SPH) with prior evidence of benefits on iron metabolism, inflammation, and metabolic health. Methods: This randomized, double-blind, active-controlled pilot study examined whether 56 days of daily ProGo® supplementation could enhance key biological drivers of healthy ageing in overweight adults, compared with an iso-nitrogenous whey protein isolate (WPI). Primary outcomes were hematological parameters and a composite integumentary (hair, nail, and skin) score, assessed with Holm–Bonferroni multiplicity adjustment. Secondary outcomes included change in body composition, inflammatory biomarkers and glucose metabolism. Results: ProGo® showed significant improvements in hemoglobin, RBC count, and ferritin, and in the integumentary composite score. Secondary analyses showed reductions in percent body fat and BMI, with preservation of lean body mass (LBM), alongside reductions in pro-inflammatory cytokines, HbA1c, and fasting blood glucose. WPI had minimal impact on any of these outcomes. Conclusions: ProGo® supplementation improved multiple key biological drivers of healthy ageing: erythropoietic function, systemic inflammation, glycemic control, and body composition. These findings are consistent with a bioactive rather than a purely nutritional effect and are hypothesis-generating with respect to a multi-domain profile relevant to healthy ageing and the maintenance of intrinsic capacity. Confirmation in larger studies is warranted.

Biomedicines doi: 10.3390/biomedicines14081662

Authors: Lanre Peter Daodu Yogini Raste Judith E. Allgrove Francesca I. F. Arrigoni Reem Kayyali

Background: Post-acute sequelae of COVID-19 (PASC), commonly known as long COVID, affects an estimated 10–30% of SARS-CoV-2 non-hospitalised and 50–70% of hospitalised survivors. This condition remains clinically heterogeneous, and the specific mechanisms driving the transition from acute infection to chronic sequelae remain poorly understood. We assessed independent risk factors, tracked the evolution of clinical features, defined distinct symptom-based phenotypes, and assessed the impact of different pandemic waves on the likelihood of developing long COVID in hospitalised survivors. Methods: We conducted a single-centre, retrospective cohort study at a university hospital in London. The population comprised 627 adults hospitalised with acute COVID-19 between February 2020 and December 2022. Baseline characteristics and outcomes were compared between long COVID and resolved cases using appropriate statistical tests for continuous and categorical variables. Multivariable logistic regression identified risk factors. McNemar’s test quantified the phenotypic shift from admission to follow-up. Latent Class Analysis (LCA) identified clinical subtypes based on symptom clusters. Results: Of 627 patients, 252 (40.2%) met long COVID criteria. Comorbidity burden was the strongest predictor; patients with a single condition had a 4-fold increase in odds (aOR 4.62, 95% CI 2.36–9.04). The ORs were also significantly elevated among patients with 2 (aOR 3.26), 3 (aOR 2.68), or 4 or more (aOR 3.24) comorbidities. Older age (aOR 1.04), acute disease severity (measured by length of hospital stay) (aOR 1.27 per log-day) and elevated admission fibrinogen (aOR 1.21 per g/L) were significant predictors. Temporal analysis revealed a precipitous decline in risk from Wild-type/Alpha (>50%) to Delta/Omicron (<21%). We observed a distinct phenotypic shift: while acute respiratory inflammation resolved, systemic fatigue increased fourfold (7.0% to 32.4%), and memory difficulties emerged in the post-acute phase. LCA identified two phenotypes: fatigue-dominant and multisystem phenotypes. Conclusions: Long COVID is a multifactorial syndrome driven by host susceptibility, acute severity, and persistent coagulopathy. Clinical management should move beyond a monolithic approach and favour phenotype-specific strategies.

Biomedicines doi: 10.3390/biomedicines14081661

Authors: Alina-Maria Ivaşko Corina Ureche Oana Cristina Marginean Monica Grama Teodora Nicola-Varo

Background: The increasing incidence of pediatric food allergy (FA) drives an imperative for mechanistically driven risk stratification beyond descriptive diagnostics. The current systematic review aims to assess the predictive and diagnostic value of early-life non-invasive biomarkers: cord blood IgE (CS-IgE), fecal calprotectin (FC), fecal zonulin (FZ) and secretory IgA (SIgA). Methods: In accordance with the PRISMA guidelines, we performed a systematic literature search on PubMed/MEDLINE and Scopus for English observational studies published in 2009–2025. The titles and abstracts were screened by two independent reviewers and methodological quality was assessed using the Newcastle–Ottawa Scale (NOS). Results: A total of 17 studies met the inclusion criteria. This synthesis demonstrates that CS-IgE reflects a systemic atopic predisposition rather than serving as a highly sensitive predictor for isolated FA. Fecal calprotectin was the most consistent marker of inflammatory activity in non-IgE-mediated food allergies, with median values in food allergy ranging from approximately 200 to 410 µg/g and proven useful in dietary monitoring. Fecal zonulin reflects epithelial barrier dysfunction; however, its clinical utility is limited by suboptimal assay specificity. SIgA primarily reflects mucosal immune maturation and tolerance development rather than serving as a direct diagnostic marker. Conclusions: Although CS-IgE, FC, FZ and SIgA exhibit potential in the risk stratification of early FA, their clinical implementation is complicated by methodological heterogeneity and concerns regarding analytical specificity, especially with respect to zonulin assays. We propose an integrated multi-biomarker risk stratification model as a conceptual hypothesis requiring prospective validation in future large-scale studies.

Biomedicines doi: 10.3390/biomedicines14081660

Authors: Licheng Xie Wenyao Jia Xitong Xu Huijuan Wu

Background: Chronic kidney disease (CKD) is a major global health burden, affecting more than 850 million people worldwide. Increasing evidence implicates engulfment and cell motility 1 (ELMO1), a cytoplasmic adaptor protein that regulates cytoskeletal remodeling, phagocytosis, and immune responses through the ELMO1–DOCK1–Rac1 signaling axis, in renal injury and disease progression. Methods: This narrative review identified relevant publications through searches of PubMed, Web of Science, and Google Scholar using combinations of the terms “ELMO1,” “kidney disease,” “diabetic kidney disease,” “IgA nephropathy,” “focal segmental glomerulosclerosis,” “acute kidney injury,” and “renal fibrosis.” English-language original research articles, genetic association studies, mechanistic investigations, and selected review articles were preferentially included according to their relevance to ELMO1 in kidney diseases. Results: Genetic association studies have implicated the ELMO1 locus in susceptibility to diabetic kidney disease (DKD), although associated variants and effect sizes differ across populations. Experimental studies suggest that ELMO1 regulates cytoskeletal remodeling, inflammatory responses, oxidative stress, and extracellular matrix deposition through the canonical ELMO1–DOCK1–Rac1 signaling pathway as well as Rac1-independent mechanisms. Available evidence supports a role for ELMO1 in DKD and renal fibrotic remodeling. In focal segmental glomerulosclerosis, the relevance of ELMO1 is primarily inferred from Rac1-associated podocyte injury pathways, whereas evidence in acute kidney injury remains limited and context-dependent. ELMO1 may contribute to inflammatory injury in ischemia–reperfusion settings but may also support efferocytosis and tissue repair in nephrotoxic injury models. In IgA nephropathy, evidence for a direct role of ELMO1 remains limited and is currently based largely on indirect mechanistic observations involving mucosal immunity and glomerular injury responses. Conclusions: ELMO1 is a context-dependent regulator of cytoskeletal, inflammatory, oxidative, and matrix-remodeling processes relevant to kidney disease. The strongest evidence currently supports its involvement in DKD, whereas its roles in other kidney diseases remain to be further defined in disease-specific, cell-type-specific, and stage-specific experimental models. Further studies are required to clarify its potential as a biomarker or therapeutic target in CKD.

Biomedicines doi: 10.3390/biomedicines14081659

Authors: Gohar Sevoyan Daniel Polianczyk Siranuysh Grabska Hovakim Grabski Ruben Abagyan Zaruhi Karabekian

Background/Objectives. Programmed death-ligand 1 (PD-L1) is a critical immune checkpoint protein that enables tumors to evade immune surveillance by suppressing T cell activation. Monoclonal antibodies are currently used to modulate PD-1/PD-L1 interactions. However, several immune-associated adverse effects were ascribed to those treatments. This led to the necessity for small-molecule alternatives like BMS-202. Methods. In this study, we investigated the temporal dynamics of cell-surface PD-L1 in response to the small-molecule dimerizer BMS-202. Treatment with a non-cytotoxic concentration of BMS-202 at 5 µM triggered a transient reduction in surface PD-L1 concentration, reaching its lowest level at 15 min. In the attempt to characterize the fate of PD-L1 following exposure to the BMS-202 dimerizer, we employed a low-pH wash internalization assay. Results. The results demonstrated a transient increase in intracellular PD-L1 within 5–15 min of compound exposure, followed by rapid recovery of surface PD-L1 levels. These findings suggest that BMS-202-induced changes in surface PD-L1 are acute and reversible, with levels returning to baseline within 24 h post-exposure. Conclusions. These findings reveal a dynamic regulatory mechanism where small-molecule-induced dimerization triggers rapid protein trafficking and transient surface depletion. Understanding these temporal dynamics is essential for the development of next-generation small-molecule immune checkpoint inhibitors.

Biomedicines doi: 10.3390/biomedicines14081658

Authors: Katharina Rump Michael Adamzik

Perioperative stress induces profound molecular and cellular responses that contribute to postoperative complications, including perioperative neurocognitive disorders (PND), chronic postsurgical pain, organ dysfunction, immunothrombosis, fibrosis, and cancer progression. Increasing evidence demonstrates that epigenetic mechanisms act as central regulators linking surgical trauma, inflammation, metabolic stress, ischemia–reperfusion injury, and immune activation to long-term alterations in gene expression and tissue remodeling. DNA methylation, histone modifications, chromatin remodeling, non-coding RNAs, and RNA epitranscriptomic mechanisms such as N6-methyladenosine (m6A) collectively orchestrate perioperative responses across multiple organ systems. Recent translational studies have identified histone deacetylases (HDACs), histone methyltransferases, NETosis-associated chromatin signaling, HMGB1/NF-κB activation, and epigenetic regulation of neuroimmune pathways as major contributors to postoperative cognitive dysfunction, chronic pain, cardiac dysfunction, pulmonary injury, and fibrosis. In parallel, advances in liquid biopsy, circulating tumor DNA (ctDNA), and single-cell epigenomics have opened new opportunities for biomarker-guided perioperative precision medicine. This review summarizes current evidence regarding epigenetic regulation in perioperative medicine with special emphasis on neuroepigenetics, NETosis, fibrosis, cardiac epigenetics, immune remodeling, and perioperative oncological outcomes. Furthermore, we discuss emerging therapeutic strategies targeting HDACs, DNA methylation, m6A pathways, and chromatin-associated inflammatory signaling as potential future interventions for perioperative complications.

Biomedicines doi: 10.3390/biomedicines14081657

Authors: Daniela Poenaru Claudia Gabriela Potcovaru Livia Alexandra Ion Andreea Dumitrescu Simona Elena Savulescu Delia Cinteza

Introduction: Chronic stroke frequently causes structural and functional impairments in the tibialis anterior (TA) muscle, leading to foot-drop and altered gait kinematics. We investigated the relationship between ultrasonographic parameters of the TA muscle and functional gait performance in chronic stroke survivors. Methods: This cross-sectional, observational pilot study evaluated eight consecutive chronic stroke patients (>6 months post-stroke). Structural parameters of TA (rest and contraction thickness, pennation angle) were documented by ultrasound imaging. A dynamic feature calculated was the Contraction Index (CI = effort/resting thickness). Functional metrics included the Medical Research Council (MRC) muscle strength scale and the 10-Meter Walk Test (10MWT) for gait velocity. Results: Statistical analysis revealed a moderate negative correlation trend between resting TA muscle thickness and functional gait speed (r = −0.618, p = 0.102). Conversely, a moderate positive correlation trend was found (r = 0.548, p = 0.160). Ultrasound imaging successfully differentiated three distinct pathological phenotypes: an atrophic phenotype (low pennation angle, flaccid muscle failure), a severely shortened spastic phenotype (increased resting thickness, high pennation angle, pathological CI < 1.0), and a spastic co-contraction loop. Patients with the atrophic phenotype achieved high mechanical efficiency with an ankle–foot orthosis (AFO), whereas the spastic phenotype exhibited resistance against the orthotic device. Conclusions: Musculoskeletal ultrasound provides objective parameters for post-stroke TA muscle remodeling and contributes to completing the assessment and therapy. Identifying specific structural muscle phenotypes allows clinicians to optimize target-specific neurorehabilitation strategies, predict AFO efficiency, and guide antispastic interventions such as botulinum toxin injections.

Biomedicines doi: 10.3390/biomedicines14081656

Authors: Arzu Dinç Yavaş Akasya Şaipi Sümeyye Akçay

Background/Objectives: Fibromyalgia (FM) is a multifactorial disorder characterized by chronic pain, fatigue, asthenia, and sleep disturbances. FM may have effects on the eyes, such as dry eye disease (DED) and retinal changes. The aim of this study is to evaluate these ocular changes in FM patients. Methods: It is a single-center prospective study. Forty-two female patients with FM (Group 1) and 37 healthy controls (Group 2) participated in the study. Study participants underwent screening tests for DED and the retinal nerve fiber layer (RNFL) thickness. Results: According to the Schirmer test, Group 1 had lower scores with a significant difference (8.2 ± 4.36 mm vs. 17.51 ± 2.41 mm). When the tear film breakup time (TBUT) test values were analyzed, there was a significant difference between groups; Group 1 had lower scores (4.95 ± 2.26 s vs. 9.86 ± 1.64 s). When the RNFL thickness values were analyzed by quadrants, our results in Group 1 FM patients were lower in the superior temporal quadrant (117.98 ± 13.98 µm vs. 124.35 ± 13.71 µm). Conclusions: These findings indicate that patients with fibromyalgia may have increased dry eye findings and localized RNFL thinning, suggesting that fibromyalgia may be associated with both anterior and posterior segment ocular alterations. However, these parameters should be interpreted as ocular manifestations rather than diagnostic markers. Further studies are needed to determine their clinical relevance and potential biomarker value.

Biomedicines doi: 10.3390/biomedicines14081655

Authors: Milan Lackovic Dejan Nikolic Jovana Kuzmanovic Pficer Sladjana Mihajlovic

Background: Migraine has been associated with vascular dysfunction and adverse pregnancy outcomes. However, the relationship between changes in migraine manifestations during pregnancy and fetal growth remains incompletely understood. This prospective study aimed to investigate whether migraine symptom progression during pregnancy is associated with abnormal fetal growth patterns and placental-mediated pregnancy complications among women with pre-existing migraine. Methods: This prospective cohort study included 400 pregnant women with a pre-pregnancy diagnosis of migraine who received antenatal care at the Department of Obstetrics and Gynecology, University Hospital “Dr Dragiša Mišović”, Belgrade, Serbia, between 2024 and 2026. Maternal migraine characteristics, pregnancy complications, and fetal growth assessments were evaluated. Fetal growth was categorized according to estimated fetal weight percentiles adjusted for gestational age. Univariable and multivariable logistic regression analyses were performed to identify factors associated with abnormal fetal growth. Because of the relatively small number of events, Firth’s penalized logistic regression was additionally performed as a sensitivity analysis. Results: Persistent or worsening migraine symptoms during pregnancy were more frequently observed among pregnancies complicated by impaired fetal growth, pregnancy-induced hypertension, gestational diabetes mellitus, abnormal gestational weight gain, and preterm birth. In univariable analyses, migraine worsening was associated with increased odds of abnormal fetal growth. However, after adjustment for maternal and obstetric factors, migraine worsening was no longer independently associated with abnormal fetal growth. Pregnancy-induced hypertension and inadequate gestational weight gain remained the strongest independent predictors. Findings were confirmed in sensitivity analyses using Firth’s penalized logistic regression. Conclusions: Among women with pre-existing migraine, persistent or worsening migraine manifestations during pregnancy were associated with a higher burden of placental-mediated complications and abnormal fetal growth patterns in unadjusted analyses. However, migraine worsening was not independently associated with abnormal fetal growth after adjustment for maternal and obstetric factors, suggesting that migraine symptom progression may represent a clinical marker of shared vascular, inflammatory, endothelial, and metabolic processes rather than an independent causal factor. Further prospective multicenter studies incorporating non-migraine control groups and detailed migraine phenotyping are warranted.

Biomedicines doi: 10.3390/biomedicines14081654

Authors: I-Min Su Shih-Yuan Ye Jer-Chuan Li Du-An Wu Bang-Gee Hsu

Background/Objectives: Endothelial dysfunction represents an early stage of vascular injury in type 2 diabetes mellitus (T2DM) and contributes to increased cardiovascular risk. Leptin, a hormone involved in metabolic and inflammatory regulation, has been implicated in vascular dysfunction. However, its association with digital thermal monitoring (DTM)-derived peripheral vascular reactivity in T2DM remains unclear. Methods: This cross-sectional study enrolled 88 patients with T2DM to investigate the association between serum leptin levels and peripheral vascular reactivity, assessed using the digital thermal monitoring-derived vascular reactivity index (VRI). Based on the VRI values, patients were categorized as having good (VRI ≥ 2.0), intermediate (VRI of 1.0–1.9), or poor (VRI < 1.0) vascular reactivity. Serum leptin levels were quantified using an enzyme immunoassay. Results: Patients with poor vascular reactivity were older and had higher total cholesterol, triglyceride, fasting glucose, glycated hemoglobin, urine albumin-to-creatinine ratio, and leptin levels. In the primary parsimonious multivariable logistic regression model adjusted for age, sex, body mass index (BMI) and eGFR, higher serum leptin levels were associated with vascular reactivity dysfunction. Linear regression analysis also showed that log-transformed leptin levels were negatively associated with VRI. Exploratory analyses, including poor vascular reactivity models and penalized regression models, showed generally consistent findings but were interpreted cautiously because of the limited number of outcome events. Conclusions: Higher serum leptin levels were associated with impaired DTM-derived peripheral vascular reactivity in clinically stable patients with T2DM after adjustment for age, sex, BMI, and eGFR. These findings should be interpreted cautiously and require validation in larger prospective studies.

Biomedicines doi: 10.3390/biomedicines14081653

Authors: Yuetong Leona Ding Dominika Bernath-Nagy Chiara Heß Florian Leuschner Hugo Albert Katus Norbert Frey Jona Benjamin Krohn Evangelos Giannitsis

Background/Objectives: High-sensitivity cardiac troponin (hs-cTn) assays are used in routine diagnostics to detect myocardial injury. However, a fraction of circulating cardiac troponin T (cTnT) enclosed within extracellular vesicles (EVs) goes widely undetected. This study introduces a combined lysis- and sonication-based protocol to release and quantify EV-bound cTnT in a time-efficient manner using a state-of-the-art hs-cTnT immunoassay. Methods: Plasma samples from patients with non-ST-segment elevation myocardial infarction (NSTEMI), unstable angina, pulmonary embolism, decompensated aortic stenosis, atrial fibrillation, myocarditis, and healthy controls were treated with a lysis buffer and subsequently sonicated. Treated and untreated samples were assessed and compared to a conventional EV isolation method. Results: Following combined lysis and sonication, cTnT levels were significantly higher compared to native, unprocessed samples across all cohorts. The median increase post-processing ranged from ~10% in decompensated aortic stenosis to ~34% in young healthy controls. In NSTEMI, the EV-bound cTnT accounted for ~15% of plasma cTnT and remained stable over 72 h. The EV cTnT/plasma cTnT ratios were comparable between the combined lysis and sonication approach and the EV isolation method. Processing time prior to cTnT measurement was reduced from ~2.5 h to ~10 min using the lysis and sonication protocol. Conclusions: Our method allows for the rapid liberation of a previously inaccessible EV-bound fraction of cTnT without the need for time-consuming and resource-intensive EV isolation workflows and is therefore readily implementable alongside standard hs-cTnT testing. The observed EV-cTnT patterns suggest differential compartmentation of cTnT, potentially reflecting the myocardial pathophysiology underlying troponin elevation.

Biomedicines doi: 10.3390/biomedicines14071652

Authors: Marc J. Braem Muriel Lins Annelies Van Den Bergh Frans Verhelst Leslee Willes Ellen Collier

Background/Objectives: The study evaluates the effectiveness of mandibular advancement device (MAD) treatment in drug-induced sleep endoscopy (DISE) preselected adults with moderate-to-severe obstructive sleep apnea (OSA). Methods: It is a retrospective observational cohort study without a control arm/treatment, including 182 patients with an apnea–hypopnea index (AHI) ≥ 15 events/h of sleep (Level-1 polysomnography) from six Belgian hospitals. After DISE preselection, a custom-made MAD (EVO, ProSomnus, Pleasanton, CA, USA) was titrated to symptom relief or physiological limits. Level-3 home polygraphy with MAD was performed within five months. The primary outcome was change in AHI from baseline. Secondary outcomes included percentage change in AHI, change in OSA severity category, snoring loudness and Epworth Sleepiness Scale (ESS) scores. Analysis of covariance methods adjusting for baseline AHI were employed. Exploratory analyses compared outcome measures by OSA severity at baseline. ‘Success’ equaled AHI < 10 with ≥50% improvement. Results: Mean AHI significantly decreased from 24.4 to 7.1 (p < 0.0001) with a significant mean percent decrease by 68.6% (p < 0.0001), improving more (p < 0.0001) in severe cases. Mean change in snoring loudness improved significantly from a mean of 7.1 to 1.7 (p < 0.0001), improving more (p = 0.049) in moderate cases. Mean change from baseline in overall ESS score was −1.7 (p < 0.0001). Greater improvements were observed in participants with elevated baseline ESS scores. MAD significantly (p = 0.03) improved OSA category: moderate OSA cases improved by a mean of 1.4 levels, severe cases by a mean of 2.1 levels (p < 0.0001). A total of 75.8% of participants achieved ‘Success’. Conclusions: In this retrospective multicenter cohort, MAD therapy was associated with significant reductions in AHI, snoring loudness, and ESS scores among DISE-preselected adults with moderate-to-severe OSA. Because treatment outcomes were assessed using Level-3 home polygraphy, the findings should be interpreted as real-world effectiveness outcomes obtained within the Belgian clinical care pathway.

Biomedicines doi: 10.3390/biomedicines14071651

Authors: Ayben Erkan Ayca Irgit Calayir Halilibrahim Ciftci Belgin Sever

Background/Objectives: Chronic myeloid leukemia (CML) is a hematological malignancy driven by the constitutively active BCR-ABL1 fusion protein. Although current ABL1 tyrosine kinase inhibitors (TKIs) have improved CML management, resistance remains a major challenge, highlighting the need for novel therapeutic strategies. Methods: A chalcone derivative containing naphthalene and toluene moieties (A) was synthesized from 2-acetonaphthone and p-tolualdehyde and subsequently converted into a novel 4,5-dihydro-1H-pyrazole-1-carbothioamide derivative (B) through reaction with thiosemicarbazide. The cytotoxicity of compounds A and B against K562 CML cells was evaluated using the MTT assay. The active compound was further investigated for cytotoxic selectivity using HL-60 acute myeloid leukemia (AML) cells and healthy PBMCs. Apoptotic effects in K562 cells were analyzed using Annexin V/ethidium homodimer staining, whereas ABL1 inhibitory activity was determined using the ADP-Glo kinase assay. The potential interaction between the active compound and ABL1 was assessed by molecular docking analysis. Results: Compound B displayed potent cytotoxic activity against K562 cells with an IC50 value of 6.92 ± 1.14 µM and demonstrated selectivity toward leukemic cells over PBMCs (SI = 5.4). Treatment with compound B markedly induced apoptosis in K562 cells. In addition, compound B inhibited ABL1 activity in a concentration-dependent manner. Molecular docking studies revealed a favorable binding orientation within the ATP-binding pocket of ABL1. Furthermore, in silico absorption, distribution, metabolism, and excretion (ADME) analysis predicted favorable pharmacokinetic properties for compound B. Conclusions: These findings demonstrate that compound B possesses cytotoxic, pro-apoptotic, and ABL1 inhibitory activities against CML cells and may serve as a promising lead structure for the development of novel therapeutic agents targeting CML.

Biomedicines doi: 10.3390/biomedicines14071650

Authors: Oriana-Eliana Pelineagră Ioana Golu Melania Balaș Daniela Georgiana Amzăr Iulia Plotuna Oana Popa Diana Aruncutean Dan Cristian Roşu Ion Icma Agneta Maria Pusztai Mărioara Cornianu Mihaela Iacob Nicu Olariu Mihaela Vlad

Background: Accurate preoperative localization of hyperfunctioning parathyroid glands remains challenging, particularly in secondary hyperparathyroidism where multiglandular disease may compromise scintigraphic performance. Methods: Our study evaluated biochemical, morphometric, and histopathological predictors of 99mTc-sestamibi scintigraphy detectability in both primary and secondary hyperparathyroidism. The study group included a total of 162 patients with primary and secondary hyperparathyroidism who underwent dual-phase 99mTc-sestamibi scintigraphy followed by parathyroidectomy. Demographics, biochemical parameters, histopathological features, lesion volume and scintigraphic findings were assessed at patient and lesion level. Results: In primary hyperparathyroidism, adenomas were larger and more frequently detected than hyperplastic glands. Lesion volume and solid growth pattern were found as positive predictors of sestamibi uptake. In secondary hyperparathyroidism, nodular hyperplasia was associated with larger volume, higher cellularity, and more frequent localizing studies. Upper quadrant position and diffusely hyperplastic lesions were associated with higher lesion miss rates, while lesion volume increased the likelihood of detection. Conclusions: Our findings highlight that 99mTc-sestamibi scintigraphy performance is strongly influenced by lesion volume, histopathological architecture and anatomical position, underscoring the needs for cautious interpretation of negative or incomplete scans, especially in secondary hyperparathyroidism.

Biomedicines doi: 10.3390/biomedicines14071648

Authors: Jessica Zablocki da Luz Leonardo Vinícius Barbosa Thiago Rodrigues dos Santos Aliciane de Almeida Roque Camila Confortin Amanda Beatriz Soares Fulan Lúcia de Noronha Deisy Morselli Gysi Cleber Machado-Souza

Background/Objectives: Rhabdomyosarcoma (RMS), the most common pediatric soft tissue sarcoma, includes two main subtypes, embryonal (eRMS) and alveolar (aRMS), each with distinct molecular and clinical characteristics. Although cellular processes underlying RMS and differences between PAX3-FOXO1 fusion-positive and fusion-negative tumors are well known, the contribution of long noncoding RNAs (lncRNAs) remains poorly understood. Methods: Here, we reannotated publicly available microarray datasets to comprehensively profile lncRNA expression and reconstruct lncRNA–miRNA–mRNA regulatory networks in RMS. Results: We identified several lncRNAs with subtype-specific differential expression, including HOTAIR as a potential sponge for miR-206, DSCR8 for miR-885-5p, and PRKCQ-AS1 for miR-515-5p in eRMS. Database-supported interaction analyses identified putative regulatory relationships between these lncRNAs and cancer-related miRNAs and mRNAs. Validation using the St. Jude Cloud PeCan platform confirmed distinct lncRNA expression signatures across RMS subtypes and other pediatric solid tumors, supporting subtype-specific regulation. Conclusions: Our findings provide an updated characterization of the lncRNA landscape in RMS and identify candidate lncRNA–miRNA–mRNA regulatory networks that may contribute to disease biology. The proposed regulatory interactions are hypothesis-generating and require experimental validation. Overall, our findings provide a resource for future functional studies and support the investigation of lncRNAs as potential biomarkers and therapeutic targets in RMS.

Biomedicines doi: 10.3390/biomedicines14071649

Authors: Bawo Ikolo Mathew Oyelami Odinaka Mgbeke Kwami Jones Shellon Thomas Felicia Ikolo

Sickle Cell Disease (SCD) is a hereditary hemoglobinopathy arising from a single-nucleotide transversion (GAG → GTG) at codon six of the HBB gene on chromosome 11, substituting glutamic acid with valine in the β-globin chain and producing hemoglobin S (HbS). Under hypoxic conditions, HbS polymerizes and distorts erythrocytes into the characteristic sickle shape, initiating a cascade of vaso-occlusion, chronic hemolytic anemia, and progressive multi-organ damage that defines the clinical burden of this disease. Although SCD has ancient origins in sub-Saharan Africa, the Indian subcontinent, the Middle East, and the Mediterranean, regions where it conferred heterozygous resistance to malaria, the ease of human migration has long since made it a global health concern, affecting an estimated 300,000–400,000 newborns annually. Advances in molecular and genomic research have deepened our understanding of SCD pathophysiology, revealing the central contributions of hemoglobin polymerization, oxidative stress, endothelial inflammation, and nitric oxide depletion to disease progression. Current management rests on supportive pharmacological interventions, including hydroxyurea, chronic transfusion therapy, L-glutamine, and multimodal pain management, complemented by lifestyle modifications. Curative approaches have advanced substantially: hematopoietic stem cell transplantation (HSCT) remains the established standard of cure, while the regulatory approvals in late 2023 of the CRISPR/Cas9-based exagamglogene autotemcel (Casgevy) and the lentiviral vector-based lovotibeglogene autotemcel (Lyfgenia) represent the most transformative development in the history of SCD therapeutics. This review traces the disease from its ancient origins and molecular characterization through to its clinical manifestations, inheritance patterns, screening strategies, and the full spectrum of current and emerging therapies. Persistent challenges, prohibitive treatment costs, healthcare inequities, the ethical dimensions of genome editing, and the urgent need for long-term safety data, are examined critically, with a view to informing the research and policy agenda that must accompany these remarkable scientific advances.

Biomedicines doi: 10.3390/biomedicines14071647

Authors: Laura M. Tsujikawa Agostina Carestia Sylwia Wasiak Christopher D. Sarsons Ravi Jahagirdar Salman Azhar Dean Gilham Derek Li Li Fu Jan O. Johansson Norman C. W. Wong Michael Sweeney Ewelina Kulikowski

Background/Objectives: Obesity can cause metabolic disorders and hepatic steatosis. Continuous hepatic influx of dietary lipids leads to adaptive metabolic changes, including increased fatty acid oxidation (FAO) and ketogenesis. However, these adaptations are not enough to counter the detrimental accumulation of lipids, resulting in hepatic steatosis. Apabetalone is a clinical-stage Bromodomain and Extra-Terminal domain inhibitor (BETi) that attenuated the increase in hepatic fibrosis score (FS) and reduced the rate of ischemic major adverse cardiovascular events and hospitalizations for heart failure in a subgroup of patients having a high likelihood of Metabolic Dysfunction-Associated Steatotic Liver Disease (MASLD) in the phase 3 clinical trial, BETonMACE. Methods: To analyze apabetalone’s effects on lipid and ketone metabolism, RNA seq, Oil Red staining and triglyceride quantification were performed in livers from a mouse model of diabetes-induced obesity and ketones were measured in plasma. Results: Mice fed a high-fat diet (HFD) were obese and demonstrated liver steatosis. Apabetalone treatment maintained the beneficial metabolic adaptation induced by HFD (increased FAO) and decreased hepatic triglycerides and lipid droplets. This inhibition of lipid anabolism redirected substrates to FAO and ketogenesis, resulting in increased plasma ketones, showing for the first time the role of BETi in ketogenesis. In the heart, apabetalone treatment reduced cardiac oxidative stress and plasma NT-proBNP levels. Conclusions: Apabetalone improves hepatic lipid handling, favoring ketogenesis. As ketones have demonstrated beneficial effects on cardiac function, increased ketones induced by apabetalone may not only contribute to the observed attenuation of FS in patients, but also a reduction in cardiac events among patients with high likelihood of MASLD, as well as in the overall trial population.

Biomedicines doi: 10.3390/biomedicines14071646

Authors: Eisuke Suganuma Satoko Honda Rina Umiguchi Sayaka Ishikawa Ayako Shimamura Marina Tanaka Masashi Kyushiki Atsuko Nakazawa

Background: Patients with Kawasaki disease (KD) who develop coronary artery aneurysms (CAAs) are at increased risk of future fatal coronary events. Pharmacotherapeutic strategies to prevent coronary stenosis are still lacking. In this exploratory study, the therapeutic effect of the angiotensin receptor blocker (ARB) losartan on coronary artery (CA) stenosis was investigated in a murine model. Methods: Five-week-old male C57BL/6J mice were intraperitoneally injected with 1000 μg of Lactobacillus casei cell wall extract (LCWE) (n = 12) to induce CA stenosis. Two weeks later, the LCWE-injected mice (n = 12) were divided into two groups: six received drinking water containing losartan (100 mg/L) (LCWE+ARB), while six received normal drinking water (LCWE group). A control group (n = 5) received phosphate-buffered saline (PBS) instead of LCWE. Sixteen weeks after LCWE administration—corresponding to the peak of CA stenosis and 14 weeks after treatment initiation—the mice were euthanized for histological evaluation of the coronary arteries. Results: Losartan treatment significantly reduced the coronary arteritis score (median [IQR (interquartile range)]: 0 [0–9.5] vs. 21.5 [15–24.3], p = 0.003). LCWE-induced neointimal formation with vascular smooth muscle cell proliferation and subsequent CA stenosis were markedly attenuated in losartan-treated mice (25% vs. 100%, p < 0.001). Losartan-associated attenuation of CA stenosis was accompanied by the preservation of medial calponin expression, a reduction in the number of proliferating cell nuclear antigen (PCNA)-positive cells in the neointima, and a decrease in serum MMP-9 (matrix metalloproteinase-9) levels. Conclusions: These findings from a murine model of KD provide preliminary evidence that losartan may attenuate coronary artery remodeling. Further mechanistic studies are warranted to clarify its potential translational relevance.

Biomedicines doi: 10.3390/biomedicines14071645

Authors: Kamile Merve Bircan Abdulkerim Elmas Mustafa Akçam

Objectives: Pediatric lower gastrointestinal bleeding encompasses a broad and age-dependent spectrum of conditions, ranging from benign anorectal disorders to inflammatory bowel disease, colorectal polyps, and less common structural or vascular lesions. Despite advances in diagnostic evaluation, contemporary pediatric data on the distribution of underlying diagnoses, particularly according to age and sex, remain limited. This study aimed to evaluate the endoscopic and histopathologic spectrum of pediatric lower gastrointestinal bleeding and to examine age- and sex-related diagnostic patterns. Methods: This retrospective single-center study included children aged 0–18 years who presented with overt lower gastrointestinal bleeding and underwent colonoscopic evaluation at a tertiary pediatric gastroenterology center between May 2010 and May 2026. Demographic data, colonoscopic and histopathologic findings, final diagnoses, inflammatory bowel disease subtypes, and polyp histopathology were reviewed. Patients were categorized into four age groups: 0–3, 4–6, 7–12, and 13–18 years. Results: A total of 268 children were included. The mean age was 11.4 ± 4.9 years, and 130 patients were female (48.5%). The most common diagnosis was inflammatory bowel disease, identified in 79 patients (29.5%), including 62 with ulcerative colitis and 17 with Crohn’s disease. Normal colonoscopic and histopathologic findings were observed in 65 patients (24.3%), anal fissure in 63 (23.5%), and polypoid lesions in 30 (11.2%). Juvenile polyp was the most common polyp subtype. Diagnostic distribution differed significantly across age groups (p < 0.001). Inflammatory bowel disease was more frequent among adolescents, whereas polypoid lesions were most common in children aged 4–6 years. Exploratory sex-based analyses suggested a female predominance in ulcerative colitis and a male predominance in Crohn’s disease and polypoid lesions. Conclusions: In children undergoing colonoscopic evaluation for lower gastrointestinal bleeding, inflammatory bowel disease, normal findings, anal fissure, and polypoid lesions were the leading diagnostic categories. Age- and sex-related patterns may help guide clinical evaluation and interpretation of colonoscopic and histopathologic findings.

Biomedicines doi: 10.3390/biomedicines14071644

Authors: Sandro La Vignera Rosita A. Condorelli

Semaglutide and tirzepatide, glucagon-like peptide-1 (GLP-1) and dual glucose-dependent insulinotropic polypeptide (GIP)/GLP-1 receptor agonists, respectively, have revolutionized the management of type 2 diabetes mellitus and obesity. Beyond their established metabolic indications, emerging preclinical and clinical evidence suggests these incretin-based therapies exert pleiotropic effects across multiple organ systems through mechanisms extending beyond glycemic control and weight reduction. This comprehensive review synthesizes current evidence for unconventional applications of semaglutide and tirzepatide across five distinct therapeutic domains: oncology, psychiatry and addiction medicine, orthopedics, aesthetic medicine, and human reproduction. In oncology, both agents demonstrate antitumor activity primarily through immune and metabolic reprogramming rather than direct cytotoxicity, with promising signals in pancreatic, thyroid, breast, and colorectal cancers. In psychiatry, modulation of mesolimbic dopamine reward pathways and GABAergic neurotransmission underlies robust preclinical and observational evidence for reducing alcohol use disorder, substance use, and binge-eating behaviors. Orthopedic applications include clinically meaningful improvements in knee osteoarthritis pain and function, though bone health signals remain mixed. Aesthetic medicine faces the dual challenge of managing GLP-1 receptor agonist-associated facial volume loss while exploring therapeutic potential in inflammatory dermatoses. In reproductive medicine, metabolic improvements translate to benefits in polycystic ovary syndrome and male fertility, though periconception safety concerns persist. Across all domains, evidence derives predominantly from preclinical models, observational cohorts, and pharmacoepidemiologic studies, with randomized controlled trials remaining limited. This review critically evaluates mechanisms, efficacy signals, safety considerations, and research priorities for each application domain, providing a roadmap for translating unconventional uses of semaglutide and tirzepatide into evidence-based clinical practice.

Biomedicines doi: 10.3390/biomedicines14071643

Authors: Qiu Li Mei Yang Yingyu Luo Nannan Zhang

Diabetic kidney disease (DKD) is a leading cause of end-stage renal disease (ESRD). Early diagnosis is therefore critical for improving patient outcomes.However, traditional clinical biomarkers are constrained by diagnostic latency and limited sensitivity, particularly in cases of non-albuminuric DKD. To address these limitations, this review systematically explores the technical framework of the pathology-anchored strategy and proposes a two-phase translational approach, consisting of Pathology Anchoring Discovery and Prospective Early Validation. This strategy employs renal biopsy as the pathological gold standard in conjunction with multi-omics technologies to correlate circulating or urinary molecules with specific renal histological lesions, ultimately identifying non-invasive biomarkers with definitive pathological relevance. While numerous biomarkers demonstrate early warning potential in high-risk populations with normal conventional indicators, the pathology-anchored framework serves as a critical bridge linking these clinical biomarkers and distinct pathological changes. This review presents potential insights for early identification, risk stratification, and prognostic assessment of DKD.