Biomedicines doi: 10.3390/biomedicines14040934

Authors: Sanyukta Bihari Dia Yang Devarshi Mukherji Aya Haggiagi

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

Biomedicines doi: 10.3390/biomedicines14040933

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

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

Biomedicines doi: 10.3390/biomedicines14040932

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

In the original publication [...]

Biomedicines doi: 10.3390/biomedicines14040931

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

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

Biomedicines doi: 10.3390/biomedicines14040930

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

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

Biomedicines doi: 10.3390/biomedicines14040929

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

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

Biomedicines doi: 10.3390/biomedicines14040928

Authors: Martina Perše

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

Biomedicines doi: 10.3390/biomedicines14040927

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

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

Biomedicines doi: 10.3390/biomedicines14040926

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

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

Biomedicines doi: 10.3390/biomedicines14040925

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

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

Biomedicines doi: 10.3390/biomedicines14040924

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

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

Biomedicines doi: 10.3390/biomedicines14040917

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

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

Biomedicines doi: 10.3390/biomedicines14040923

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

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

Biomedicines doi: 10.3390/biomedicines14040921

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

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

Biomedicines doi: 10.3390/biomedicines14040922

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

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

Biomedicines doi: 10.3390/biomedicines14040920

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

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

Biomedicines doi: 10.3390/biomedicines14040919

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

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

Biomedicines doi: 10.3390/biomedicines14040918

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

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

Biomedicines doi: 10.3390/biomedicines14040916

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

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

Biomedicines doi: 10.3390/biomedicines14040915

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

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

Biomedicines doi: 10.3390/biomedicines14040914

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

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

Biomedicines doi: 10.3390/biomedicines14040913

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

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

Biomedicines doi: 10.3390/biomedicines14040912

Authors: Yujiang Luo Yaxiong Deng Li Yuan Siqi Fu

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

Biomedicines doi: 10.3390/biomedicines14040910

Authors: Lingzhi Peng Na Sun Biqiong Ren

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

Biomedicines doi: 10.3390/biomedicines14040911

Authors: John K. Triantafillidis

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

Biomedicines doi: 10.3390/biomedicines14040907

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

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

Biomedicines doi: 10.3390/biomedicines14040909

Authors: Veronika A. Prikhodko Sergey V. Okovityi

Metabolic dysfunction-associated steatotic liver disease (MASLD) is characterized by an exceptionally high global prevalence that is projected to continue rising in the near future. MASLD is strongly associated with a spectrum of cardiometabolic risk factors, and may itself, in turn, contribute to cardiovascular morbidity and mortality. This interconnection warrants the development of integrated treatment strategies targeting shared pathophysiological processes and addressing both hepatic, metabolic, and cardiovascular outcomes. In this work, we review the modern MASLD clinical development pipeline and highlight the most prominent drug candidates with known or purported cardiovascular benefits, discussing mechanistic links and supporting evidence ranging from preclinical experiments to real-world data. Although the drug development pipeline is extensive and diverse, evidence supporting cardiovascular benefits for most candidate molecules remains limited. Both of the FDA-approved therapies, resmetirom and semaglutide, have been found to significantly reduce the risk of major adverse cardiovascular events as well as cardiovascular and all-cause mortality in patients with MASH. In addition, significant improvements were observed in patients with heart failure with preserved ejection fraction treated with semaglutide, highlighting incretin mimetics as a promising class for managing cardiovascular disease concomitant with MASLD/MASH. Other investigational compounds, targeting the farnesoid X receptor, peroxisome proliferator-activated receptors, de novo lipogenesis enzymes, and fibroblast growth factors, have demonstrated improvements in blood lipid spectrum and glycemic control; however, their clinical effectiveness in patients at cardiovascular risk has yet to be established.

Biomedicines doi: 10.3390/biomedicines14040908

Authors: Nikolaos Martinos Christos Kroupis Andreas C. Lazaris Georgia-Eleni Thomopoulou

Background/Objectives: Deep remission, defined as the coexistence of endoscopic remission and histologic healing, has emerged as an advanced therapeutic target in ulcerative colitis (UC). However, reliable non-invasive biomarkers capable of accurately reflecting histologic inflammatory status remain limited. We aimed to evaluate the association between serum interleukin-10 (IL-10) concentrations and deep remission in UC and to assess its discriminatory performance. Methods: In this prospective single-center observational study, consecutive adult patients with ulcerative colitis undergoing clinically indicated colonoscopy were enrolled. Serum IL-10 concentrations were measured prior to endoscopy using an enzyme-linked immunosorbent assay. Histologic healing was defined as a Geboes score < 2.0, and deep remission as the coexistence of endoscopic remission and histologic healing. Associations were evaluated using Firth penalized logistic regression. Discrimination was assessed using receiver operating characteristic analysis with bootstrap internal validation (500 resamples). Results: Among 44 patients, 18 (40.9%) achieved deep remission. Serum IL-10 levels were significantly higher in patients with histologic healing compared with those with active histologic inflammation (p < 0.001). Log-transformed IL-10 was independently associated with deep remission in both univariable and multivariable models. The primary multivariable model demonstrated apparent good discrimination (apparent AUC 0.88; optimism-corrected AUC 0.85). Among patients in endoscopic remission, non-detectable IL-10 identified persistent histologic activity with high sensitivity (92.9%) and negative predictive value (94.4%), although these findings are exploratory. Conclusions: Serum IL-10 concentrations were independently associated with deep remission and showed potentially promising internally validated discriminatory performance within this cohort. These findings are hypothesis-generating and require external validation in larger cohorts before clinical application.

Biomedicines doi: 10.3390/biomedicines14040906

Authors: Berna Yıldırım Burcu Biltekin Mete Hakan Karalök Ayhan Bilir

Background/Objectives: Endometrial cancer frequently develops resistance to apoptosis-based therapies, highlighting the need for alternative strategies that control tumor growth independently of cell death induction. Three-dimensional (3D) tumor models more accurately recapitulate tumor architecture, cellular interactions, and treatment resistance compared to conventional two-dimensional (2D) cultures. This study aimed to investigate whether imatinib-based combination treatments can enforce sustained cytostatic responses in a 3D endometrial cancer model. Methods: Ishikawa spheroids were treated with imatinib alone or in combination with lithium chloride or medroxyprogesterone acetate. Proliferation was assessed by bromodeoxyuridine incorporation, cell cycle distribution by flow cytometry, and apoptosis by Annexin V/propidium iodide staining over 96 h. Results: Imatinib monotherapy produced modest antiproliferative effects, whereas combination treatments resulted in sustained suppression of DNA synthesis, increased G0/G1 accumulation, and reduced S-phase entry. Despite strong growth inhibition, apoptotic fractions remained low across all groups. Conclusions: Imatinib-based combinations suppress 3D endometrial cancer growth predominantly through sustained cell cycle arrest rather than apoptosis induction. Targeting apoptosis-resistant proliferation through cytostatic mechanisms may represent a complementary therapeutic strategy for hormone-responsive endometrial cancer and warrants further translational evaluation.

Biomedicines doi: 10.3390/biomedicines14040904

Authors: Jinjing Huang Dapeng Wang Chengyun Xu Jianping Wu

Background: Dodder, the dried mature seed of Cuscuta chinensis Lam. (CCL), has demonstrated anti-tumor activity, but its molecular and immunological mechanisms in clear cell renal cell carcinoma (ccRCC) remain unclear. Objective: To identify potential targets and elucidate the immune mechanisms by which CCL exerts therapeutic effects against ccRCC. Methods: A network pharmacology approach was employed to predict CCL’s bioactive components and their putative targets in ccRCC. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were used to explore relevant pathways. Molecular docking validated the binding of key compounds to hub proteins. In vitro assays—including cell viability, colony formation, invasion, and apoptosis measurements—assessed the effects of quercetin, a principal CCL constituent, on 786-O renal carcinoma cells. Flow cytometry was performed to determine the percentage of CD163+ cells. An in vivo xenograft model evaluated CCL’s anti-tumor efficacy. Western blotting, flow cytometry, and multiplex immunohistochemistry (mIHC) examined the modulation of signaling pathways and immune cell markers. Results: Network pharmacology identified IL-6, EGFR, TLR4, MMP9, CD44, and IFN-γ as core targets of CCL in ccRCC. Enrichment analyses implicated immune regulation, inflammation modulation, and PI3K/AKT signaling inhibition. Molecular docking revealed strong quercetin–MMP9 binding affinity. Immuno-correlation analyses indicated that high MMP9 levels positively correlated with macrophage infiltration and M2 polarization, suggesting a role in tumor immune escape. Quercetin significantly reduced the viability of 786-O cells in a dose-dependent manner, showing approximately 45% inhibition at 80 μM (p < 0.01). In addition, quercetin decreased MMP9 expression and reduced the proportion of CD163-positive macrophages. These effects were reversed by FSL-1 TFA (Toll-like receptor 2/6 agonist), which is the agonist of MMP-9. In the xenograft model, tumor volume in the quercetin-treated group was reduced by approximately 50% compared with the control group. Conclusions: CCL, particularly its active component quercetin, may inhibit ccRCC progression via inhibiting MMP9-mediated M2 macrophage polarization.

Biomedicines doi: 10.3390/biomedicines14040905

Authors: Juraj Jug Martina Lovrić Benčić Tomislav Bulum Ingrid Prkačin

Background: R wave amplitude in the aVL ECG lead (RaVL) has been identified as a marker of cardiovascular (CV) risk, hypertension-mediated target organ damage (HMOD), and mortality in patients with arterial hypertension (AH), where RaVL > 1.1 mV suggests left ventricular hypertrophy. However, the exact threshold for identifying high-risk patients has yet to be determined. Therefore, we compared RaVL values among hypertensive patients with and without hypertensive urgencies (HUs) and healthy subjects, aiming to identify the predictors of elevated RaVL and to compare its prognostic value with the SCORE 2 model. Methods: This cross-sectional study included 339 participants divided into three groups according to ambulatory blood pressure monitoring results: 100 patients with AH and HU from the emergency department, 134 patients with AH without HU, and 105 healthy subjects recruited from four family medicine practices. Basic laboratory parameters were determined, SCORE 2 risk was calculated, PWV was measured using oscillometry, and a standard 12-lead ECG was recorded in all participants. Results: Participants with AH and HU had the highest RaVL values compared to those with AH without HU and healthy subjects (averages of 0.76 ± 0.24 mV, 0.49 ± 0.27, 0.22 ± 0.25, respectively; p < 0.001). Significantly higher RaVL values were observed in males compared to females (0.56 ± 0.31 vs. 0.41 ± 0.34 mV; p < 0.001) and in non-dippers compared to dippers (0.56 ± 0.34 mV vs. 0.41 ± 0.31 in dippers; p < 0.001). Age, mean arterial pressure, PWV, and SCORE 2 were shown as independent predictors of RaVL. Compared with SCORE 2, individuals with RaVL > 0.40 mV had high CV risk (sensitivity of 58.16%, specificity of 73.68%; p < 0.001). Conclusions: In this study, RaVL demonstrated good prognostic value for CV risk stratification. However, larger studies are needed to determine a precise high-risk threshold to improve CV risk estimation and HMOD detection in patients with marginal SCORE 2 CV risk.

Biomedicines doi: 10.3390/biomedicines14040903

Authors: Tim Rozovsky Adrian Siapno David Y. C. Cheung Sara M. Telles-Langdon Allison Ledingham Paris R. Haasbeek Lauren Castagna Lana Mackic Leah Schwartz James A. Thliveris Danielle Desautels Joerg Herrmann Davinder S. Jassal

Background/Objectives: While doxorubicin (DOX) and trastuzumab (TRZ) improve overall survival in women with breast cancer, these two anti-cancer drugs increase the risk of developing heart failure. As a novel and largely unexplored approach, our aim was to evaluate whether the prophylactic use of the sodium-glucose co-transporter 2 inhibitor empagliflozin (EMPA), with and without the angiotensin converting enzyme inhibitor perindopril (PER), is cardioprotective in preventing DOX + TRZ-mediated cardiotoxicity. Methods: In a chronic in vivo murine model, female mice received prophylactic treatment with PER (3 mg/kg), EMPA (10 mg/kg), or EMPA + PER via oral gavage for a total of 3 weeks as a run-in period prior to weekly administration of DOX + TRZ (8 mg/kg and 3 mg/kg, respectively) intraperitoneally for an additional 3 weeks (total of 6 weeks). Results: In mice treated with DOX + TRZ, the left ventricular ejection fraction (LVEF) decreased from 75 ± 2% at baseline to 40 ± 4% at week 6. Prophylactic treatment with either PER, EMPA, or EMPA+PER improved LVEF to 58 ± 3%, 66 ± 3%, and 67 ± 4% at week 6, respectively (p < 0.05). Histological analyses confirmed significant disruption of myofibrils, vacuolization, and loss of sarcomere integrity in the DOX + TRZ-treated mice. Prophylactic administration with PER, EMPA, or EMPA + PER, however, improved myofibril integrity at week 6 in mice receiving DOX + TRZ. Finally, although the Bax/Bcl-xL ratio was significantly elevated by 1.5-fold in mice treated with DOX + TRZ, this marker of apoptosis was attenuated by prophylactic treatment with either PER, EMPA, or EMPA + PER. Conclusions: Prophylactic administration of EMPA mitigated adverse cardiovascular remodeling in a chronic in vivo model of DOX + TRZ-mediated cardiotoxicity.

Biomedicines doi: 10.3390/biomedicines14040902

Authors: Valentin Bilgeri Philipp Spitaler Jasmina Gavranovic-Novakovic Theresa Dolejsi Patrick Rockenschaub Moritz Messner Marc Michael Zaruba Fabian Barbieri Agne Adukauskaite Markus Stühlinger Bernhard Erich Pfeifer Pietro Lacaita Gudrun Feuchtner Peter Willeit Axel Bauer Wolfgang Dichtl

Background: Pacemakers enable continuous long-term surveillance of atrial fibrillation detected by implanted devices. Circulating biomarkers reflecting endothelial dysfunction, inflammation, and myocardial stress may help identify patients at risk for atrial fibrillation (AF) progression and higher arrhythmic burden. Methods: This analysis included patients from the prospective ACaSA study (NCT05127720) with a dual chamber pacemaker (Microport® BOREA DR or TEO DR) and monitored weekly via remote monitoring technology (SMARTVIEW®). Individuals with permanent AF or single-chamber systems were excluded. Baseline plasma concentrations of angiopoietin-2 (ANGPT2), growth differentiation factor-15 (GDF-15), fibroblast growth factor-23 (FGF-23), bone morphogenetic protein-10 (BMP10), and tumor necrosis factor–related apoptosis-inducing ligand receptor-2 (TRAIL-R2) were quantified using enzyme-linked immunosorbent assays. N-terminal pro-B-type natriuretic peptide (NT-proBNP) was measured using electrochemiluminescence immunoassay. Biomarkers were log2-transformed, with values below assay detection limits imputed at half the lower limit of detection. Two endpoints were assessed following a 30-day blanking period: (1) progression to persistent AF, defined as ≥7 consecutive days with >99% daily AF burden, analyzed using Cox regression; and (2) AF burden, calculated as total AF time normalized to monitored days and categorized as <25%, 25–75%, or >75%, analyzed using multinomial logistic regression. Multivariable models were adjusted for age, sex, heart failure, diabetes, and prior myocardial infarction; Cox models were limited to age, sex, and heart failure due to fewer events. Results: A total of 223 patients were included (median age 75 years; 37.2% women). During follow-up, 28 patients (13.3%) progressed to persistent AF. Higher baseline ANGPT2 was the strongest predictor of progression (HR per doubling 1.83, 95% CI 1.27–2.66, p = 0.001), followed by GDF-15 (HR 1.52, 95% CI 1.03–2.24, p = 0.036). In the burden analysis, ANGPT2 demonstrated a pronounced graded relationship with arrhythmic load, with markedly increased odds of high (>75%) AF burden (OR 8.31, 95% CI 2.63–26.26, p < 0.001). GDF-15 independently predicted both medium (OR 2.05, p = 0.025) and high burden (OR 2.32, p = 0.037). NT-proBNP displayed a borderline association with high burden (OR 2.02, p = 0.061). No significant associations were observed for FGF-23, BMP10, or TRAIL-R2. Conclusions: In continuously monitored pacemaker patients, ANGPT2 and GDF-15 emerged as key biomarkers associated with AF disease severity. ANGPT2 was strongly linked to both progression to persistent AF and high AF burden, whereas GDF-15 consistently predicted higher AF burden and also contributed to risk of progression. These findings highlight endothelial and inflammatory pathways as potential markers of atrial disease progression.

Biomedicines doi: 10.3390/biomedicines14040901

Authors: Mai S. Sater Hayder A. Giha

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

Biomedicines doi: 10.3390/biomedicines14040900

Authors: Osama Al-Bataineh Rula Abdallat Ausilah Alfraihat

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

Biomedicines doi: 10.3390/biomedicines14040899

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

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

Biomedicines doi: 10.3390/biomedicines14040898

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

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

Biomedicines doi: 10.3390/biomedicines14040897

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

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

Biomedicines doi: 10.3390/biomedicines14040896

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

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

Biomedicines doi: 10.3390/biomedicines14040895

Authors: Li Wei Chunchen Gao Hongyan Qin

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

Biomedicines doi: 10.3390/biomedicines14040894

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

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

Biomedicines doi: 10.3390/biomedicines14040893

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

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

Biomedicines doi: 10.3390/biomedicines14040892

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

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

Biomedicines doi: 10.3390/biomedicines14040891

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

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

Biomedicines doi: 10.3390/biomedicines14040890

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

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

Biomedicines doi: 10.3390/biomedicines14040889

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

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

Biomedicines doi: 10.3390/biomedicines14040888

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

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

Biomedicines doi: 10.3390/biomedicines14040887

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

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

Biomedicines doi: 10.3390/biomedicines14040886

Authors: Ahmed Mansour Al Rajeh

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

Biomedicines doi: 10.3390/biomedicines14040885

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

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

Biomedicines doi: 10.3390/biomedicines14040884

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

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

Biomedicines doi: 10.3390/biomedicines14040883

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

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

Biomedicines doi: 10.3390/biomedicines14040882

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

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

Biomedicines doi: 10.3390/biomedicines14040881

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

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

Biomedicines doi: 10.3390/biomedicines14040880

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

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

Biomedicines doi: 10.3390/biomedicines14040878

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

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

Biomedicines doi: 10.3390/biomedicines14040879

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

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

Biomedicines doi: 10.3390/biomedicines14040877

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

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

Biomedicines doi: 10.3390/biomedicines14040876

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

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

Biomedicines doi: 10.3390/biomedicines14040875

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

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

Biomedicines doi: 10.3390/biomedicines14040874

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

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

Biomedicines doi: 10.3390/biomedicines14040872

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

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

Biomedicines doi: 10.3390/biomedicines14040873

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

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

Biomedicines doi: 10.3390/biomedicines14040871

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

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

Biomedicines doi: 10.3390/biomedicines14040869

Authors: Cengizhan Kurt Arif Gök

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

Biomedicines doi: 10.3390/biomedicines14040867

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

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

Biomedicines doi: 10.3390/biomedicines14040868

Authors: Fangqiao Zheng Zhengyi Lan Hangrong Chen Ming Ma

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

Biomedicines doi: 10.3390/biomedicines14040870

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

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

Biomedicines doi: 10.3390/biomedicines14040866

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

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

Biomedicines doi: 10.3390/biomedicines14040865

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

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

Biomedicines doi: 10.3390/biomedicines14040864

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

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

Biomedicines doi: 10.3390/biomedicines14040863

Authors: Weronika Rzeska Aneta Adamiak-Godlewska

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

Biomedicines doi: 10.3390/biomedicines14040860

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

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

Biomedicines doi: 10.3390/biomedicines14040862

Authors: Mehmet İncebıyık Adalet Göçmen

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

Biomedicines doi: 10.3390/biomedicines14040861

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

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

Biomedicines doi: 10.3390/biomedicines14040859

Authors: Yujia Lin Panpan Lu Qiang Ding Mei Liu

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

Biomedicines doi: 10.3390/biomedicines14040858

Authors: Xin-Fang Leong

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

Biomedicines doi: 10.3390/biomedicines14040857

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

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

Biomedicines doi: 10.3390/biomedicines14040855

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

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

Biomedicines doi: 10.3390/biomedicines14040856

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

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

Biomedicines doi: 10.3390/biomedicines14040854

Authors: John A. Dangerfield Christoph Metzner

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

Biomedicines doi: 10.3390/biomedicines14040851

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

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

Biomedicines doi: 10.3390/biomedicines14040853

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

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

Biomedicines doi: 10.3390/biomedicines14040852

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

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

Biomedicines doi: 10.3390/biomedicines14040850

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

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

Biomedicines doi: 10.3390/biomedicines14040849

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

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

Biomedicines doi: 10.3390/biomedicines14040848

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

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

Biomedicines doi: 10.3390/biomedicines14040847

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

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

Biomedicines doi: 10.3390/biomedicines14040846

Authors: Shasha Mei Hua Chen Jiezhi Dai

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

Biomedicines doi: 10.3390/biomedicines14040845

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

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

Biomedicines doi: 10.3390/biomedicines14040844

Authors: Dan Faganeli Metoda Lipnik-Stangelj

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

Biomedicines doi: 10.3390/biomedicines14040843

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

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

Biomedicines doi: 10.3390/biomedicines14040842

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

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

Biomedicines doi: 10.3390/biomedicines14040841

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

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

Biomedicines doi: 10.3390/biomedicines14040840

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

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

Biomedicines doi: 10.3390/biomedicines14040838

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

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

Biomedicines doi: 10.3390/biomedicines14040839

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

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

Biomedicines doi: 10.3390/biomedicines14040837

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

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

Biomedicines doi: 10.3390/biomedicines14040836

Authors: Vinay Devulapalli Akash Sathiyamurthi Surabhi Gautam Pallavi Bhattaram

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

Biomedicines doi: 10.3390/biomedicines14040835

Authors: Senyang Guo Jianhua Liu Hongmei Zheng Xinhong Wu

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