International Journal of Molecular Sciences

21 pages, 3916 KB

Open AccessArticle

Combination of Melittin and Clinacanthus nutans Extract Enhances Antiviral Efficacy Against Dengue Virus Serotype 2 Infection

by Natthanich Boonsatit, Somluethai Fungfueang, Saruda Thongyim, Yingmanee Tragoolpua, Terd Disayathanoowat, George S. Baillie and Aussara Panya

Int. J. Mol. Sci. 2026, 27(17), 7782; https://doi.org/10.3390/ijms27177782 (registering DOI) - 31 Aug 2026

Abstract

Dengue virus (DENV) remains a major global health threat, particularly in tropical and subtropical regions, due to the lack of effective antiviral therapies. In this study, we evaluated a novel combination strategy using melittin, a bee venom-derived antiviral peptide, together with Clinacanthus nutans [...] Read more.

Dengue virus (DENV) remains a major global health threat, particularly in tropical and subtropical regions, due to the lack of effective antiviral therapies. In this study, we evaluated a novel combination strategy using melittin, a bee venom-derived antiviral peptide, together with Clinacanthus nutans extract to enhance antiviral efficacy while reducing melittin-associated cytotoxicity against dengue virus serotype 2 (DENV-2). Cytotoxicity was assessed in Vero cells using a cell viability assay, whereas antiviral activity was evaluated by focus-forming unit (FFU) reduction assay, cell-based ELISA, FFU titration assay, and immunofluorescence assay (IFA). Melittin (1.25–2.5 µg/mL) and C. nutans extract (15.625–500 µg/mL) maintained cell viability above 80%, whereas 5 µg/mL melittin markedly reduced cell viability. Notably, co-treatment with C. nutans restored cell viability to above 80%, demonstrating a significant cytoprotective effect. More importantly, the combination treatment exhibited greater antiviral activity than either agent alone, resulting in complete inhibition of viral infection at lower concentrations while simultaneously suppressing intracellular viral protein expression and production of progeny viruses. These findings suggest that C. nutans may enhance the antiviral activity of melittin while reducing its cytotoxicity under the conditions tested. Overall, this study provides preliminary evidence supporting the potential of this natural compound-based combination strategy and warrants further preclinical investigation as a candidate antiviral approach against DENV infection. Full article

(This article belongs to the Special Issue Novel Antiviral Agents: Challenges and Opportunities from Molecular Targeting to Validation)

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17 pages, 7014 KB

Open AccessArticle

A Comparative Analysis of the Methylation Status of Non-Coding RNA Promoters in Fibroid and Matched Myometrium

by Tsai-Der Chuang, Shawn Rysling, Abigail Wiseman, Gabriela Alfaro, Sayna Pejouhesh Jahromi, Sepideh Pejouhesh Jahromi, Daniel Baghdasarian and Omid Khorram

Int. J. Mol. Sci. 2026, 27(17), 7781; https://doi.org/10.3390/ijms27177781 (registering DOI) - 31 Aug 2026

Abstract

Uterine fibroids exhibit dysregulated expression of non-coding RNAs (ncRNAs), although the underlying mechanisms remain incompletely understood. We investigated promoter DNA methylation and its relationship with ncRNA expression in fibroids. Genomic DNA from eight paired fibroid and matched myometrial tissues was analyzed using MeDIP–chip [...] Read more.

Uterine fibroids exhibit dysregulated expression of non-coding RNAs (ncRNAs), although the underlying mechanisms remain incompletely understood. We investigated promoter DNA methylation and its relationship with ncRNA expression in fibroids. Genomic DNA from eight paired fibroid and matched myometrial tissues was analyzed using MeDIP–chip to identify differentially methylated ncRNA promoters. Selected candidates were validated by methylation-specific PCR (MSP) in 16 paired samples, and transcript expression was assessed by qRT-PCR in 68–94 paired specimens. MeDIP–chip identified 538 lncRNAs and 61 miRNAs with differential promoter methylation, including 300 hypermethylated and 238 hypomethylated lncRNAs and 47 hypermethylated and 14 hypomethylated miRNAs. Promoter methylation was not significantly correlated with transcript expression (r = −0.1224). MSP confirmed hypermethylation of LINC-PINT and MIR9-3 and hypomethylation of WT1-AS and TTLL10-AS1. Correspondingly, LINC-PINT and MIR9-3 expression was decreased, whereas WT1-AS and TTLL10-AS1 expression was increased in fibroids. However, LINC-PINT and TTLL10-AS1 methylation did not fully correspond with MeDIP–chip findings. These results reveal widespread ncRNA promoter methylation alterations in uterine fibroids but demonstrate that genome-wide methylation does not consistently predict transcript expression, highlighting the complexity of ncRNA epigenetic regulation and the importance of locus-specific validation. Full article

(This article belongs to the Section Molecular Genetics and Genomics)

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19 pages, 4932 KB

Open AccessArticle

Targeting MET and mTOR Synergistically Overcomes Adaptive Resistance in Glioblastoma

by Yunzhan Li, Hanif Khan, Seyma Demirsoy, Muhammad Younis, Guilan Shi, Hannah Valensi, William Bernhardt, Jeongwu Lee, Mitchell Machtay, Dawit Aregawi, Michael Glantz, Pierre Giglio, Shengyu Yang, Todd Schell, Vonn Walter, Yasin Uzun and Inan Olmez

Int. J. Mol. Sci. 2026, 27(17), 7780; https://doi.org/10.3390/ijms27177780 (registering DOI) - 30 Aug 2026

Abstract

Glioblastoma (GBM) is the most aggressive primary brain tumor with limited therapeutic options and extremely poor prognosis. Aberrant activation of the receptor tyrosine kinase MET drives tumor progression, therapeutic resistance, and reduced survival, particularly in the mesenchymal GBM subtype. Given its crucial role [...] Read more.

Glioblastoma (GBM) is the most aggressive primary brain tumor with limited therapeutic options and extremely poor prognosis. Aberrant activation of the receptor tyrosine kinase MET drives tumor progression, therapeutic resistance, and reduced survival, particularly in the mesenchymal GBM subtype. Given its crucial role in GBM recurrence and progression, we investigated the mechanisms of resistance to MET inhibition using patient-derived glioma-initiating cells (GICs) and orthotopic xenograft mouse models. GICs were treated with the MET inhibitor crizotinib to elucidate the mechanism of adaptive resistance. Prolonged MET inhibition induced a senescent-like phenotype in GICs, associated with downregulation of BNIP3, a mitochondrial protein regulating mitophagy. We showed that BNIP3 downregulation led to activation of mTOR signaling, promoting cellular survival and adaptive resistance. Combining crizotinib with the mTOR inhibitor everolimus effectively suppressed mTOR activity, reduced cell viability, and induced mitochondrial alterations, apoptosis, and necroptosis. In orthotopic GBM xenograft models, combined MET and mTOR inhibition significantly prolonged survival compared with single-agent treatments. Notably, sequential treatment—crizotinib followed by everolimus—further enhanced therapeutic efficacy. These effects were achieved without significant weight loss, supporting tolerability of the treatment regimen. Our findings identify the BNIP3-mTOR axis as a critical mediator of resistance to MET inhibition and demonstrate that combined inhibition of MET and mTOR exhibits significant synergy against GBM. Full article

(This article belongs to the Special Issue Emerging Therapeutic Strategies for Glioblastoma)

18 pages, 13622 KB

Open AccessArticle

Metformin Suppresses Metastatic Potential and Sensitizes Ovarian Cancer Cells to Chemotherapeutics

by Megha J. Pandya, Ayokunnumi Ogunsanya, Adedoyin Ajibade and Achuth Padmanabhan

Int. J. Mol. Sci. 2026, 27(17), 7779; https://doi.org/10.3390/ijms27177779 (registering DOI) - 30 Aug 2026

Abstract

The failure of current therapeutics to elicit a durable response in metastatic ovarian cancer patients highlights the urgent need to develop more effective treatment strategies. In this study, we demonstrate that the anti-diabetic drug metformin exerts cytotoxic effects across a broad panel of [...] Read more.

The failure of current therapeutics to elicit a durable response in metastatic ovarian cancer patients highlights the urgent need to develop more effective treatment strategies. In this study, we demonstrate that the anti-diabetic drug metformin exerts cytotoxic effects across a broad panel of ovarian cancer cell lines. Transcriptomic analyses revealed that, in addition to reducing cell viability and proliferation, metformin modulates pathways associated with metastatic progression, including cell migration, extracellular matrix remodeling, and cellular responses to chemotherapeutic agents. Consistent with these molecular changes, metformin significantly impaired ovarian cancer cell migration and invasion through Matrigel. Furthermore, metformin diminished the ability of ovarian cancer cells to form multicellular aggregates, a key property that facilitates successful metastatic dissemination within the peritoneal cavity. Although metformin produced only a modest increase in carboplatin sensitivity, systematic screening of FDA-approved drugs identified several agents whose efficacy was enhanced by metformin, including drugs not currently used to treat ovarian cancer. Collectively, these findings demonstrate that metformin suppresses metastatic phenotypes and enhances therapeutic vulnerability in ovarian cancer cells, supporting its potential repurposing as a combination therapy to improve treatment outcomes in ovarian cancer. Full article

(This article belongs to the Special Issue Advances in Ovarian Cancer Metastasis and Chemotherapy Resistance)

17 pages, 6869 KB

Open AccessArticle

1-Hydroxypyrene Promotes Bladder Cancer Progression and Tumor-Associated Immunosuppression via MIF

by Yiting Liu, Zongyu Li, Yingchun Kuang, Ke Chen and Lilong Liu

Int. J. Mol. Sci. 2026, 27(17), 7778; https://doi.org/10.3390/ijms27177778 (registering DOI) - 30 Aug 2026

Abstract

This study examined how 1-hydroxypyrene (1-OHP) affects bladder cancer (BCa) progression and tumor-associated immunosuppression. Public transcriptomic data and target-prediction databases were used to screen for targets related to 1-OHP and BCa. Machine learning was then used to narrow down the candidate genes. Molecular [...] Read more.

This study examined how 1-hydroxypyrene (1-OHP) affects bladder cancer (BCa) progression and tumor-associated immunosuppression. Public transcriptomic data and target-prediction databases were used to screen for targets related to 1-OHP and BCa. Machine learning was then used to narrow down the candidate genes. Molecular docking and single-cell RNA sequencing were also used to examine the selected targets and related pathways. The role of the main target was tested in vitro. CCK-8, colony formation, EdU, and Transwell assays were used to assess cell growth, migration, and invasion. Protein expression was examined by Western blotting. A tumor cell–CD8⁺ T-cell co-culture system was used to evaluate changes in CD8⁺ T-cell function. We identified 29 targets shared by 1-OHP and BCa. Machine learning further narrowed these targets to six genes, and macrophage migration inhibitory factor (MIF) was chosen for experimental validation. Treatment with 1-OHP increased BCa cell viability, proliferation, migration, and invasion. Silencing MIF partly reduced these effects. 1-OHP also increased MIF and CD74/CD44 expression and enhanced PI3K/AKT/mTOR signaling. These changes were reduced after MIF silencing. In the co-culture system, 1-OHP-treated BCa cells reduced CD8⁺ T-cell proliferation and function. Ki67, GZMB, and IFNG levels were decreased. MIF knockdown partly reversed these changes. Collectively, our findings show that 1-OHP promotes bladder cancer progression and tumor-associated immunosuppression, at least in part, through a MIF-dependent mechanism. By linking environmental exposure to activation of the CD74/CD44–PI3K/AKT/mTOR axis and impaired CD8⁺ T-cell function, our study provides new insight into the molecular basis of environmentally associated bladder carcinogenesis. Full article

(This article belongs to the Special Issue Toxicity Mechanism of Emerging Pollutants: 2nd Edition)

18 pages, 1006 KB

Open AccessArticle

Systemic Immune-Inflammatory Biomarkers in Epithelial Ovarian Cancer: Subgroup-Dependent Prognostic Performance and Integrated Risk Stratification

by E Sun Paik, Young Eun Chung, Seoyoung Youn, Seongyun Lim, Jun-Hyeong Seo, Chel Hun Choi, Tae-Joong Kim, Jeong-Won Lee and Yoo-Young Lee

Int. J. Mol. Sci. 2026, 27(17), 7777; https://doi.org/10.3390/ijms27177777 (registering DOI) - 30 Aug 2026

Abstract

Systemic immune-inflammation index (SII), neutrophil-to-lymphocyte ratio (NLR), and platelet-to-lymphocyte ratio (PLR) are surrogate markers of the systemic immune-inflammatory response proposed as perioperative prognostic biomarkers in epithelial ovarian cancer (EOC), yet their performance across subgroups remains unexamined. In 373 EOC patients undergoing primary cytoreductive [...] Read more.

Systemic immune-inflammation index (SII), neutrophil-to-lymphocyte ratio (NLR), and platelet-to-lymphocyte ratio (PLR) are surrogate markers of the systemic immune-inflammatory response proposed as perioperative prognostic biomarkers in epithelial ovarian cancer (EOC), yet their performance across subgroups remains unexamined. In 373 EOC patients undergoing primary cytoreductive surgery, postoperative day-1 changes (ΔSII, ΔNLR, ΔPLR) were compared for overall survival (OS) and progression-free survival (PFS) across 10 subgroups, and a Clinical-Inflammatory Risk Score (CIRS) combining inflammation with stage and residual disease was developed. Individual markers showed modest discrimination (area under the curve [AUC] 0.579–0.615); the marker with the highest AUC varied by context, with ΔPLR ranking first most often, notably in non-serous tumors, though none was statistically superior. Seeking a molecular counterpart, two public transcriptomic cohorts were re-analyzed: VWF was consistently higher in clear cell than serous carcinoma, whereas IL6–STAT3-related differences were cohort-dependent. The CIRS achieved AUCs of 0.752 (OS) and 0.764 (PFS), a six-fold mortality gradient (7.2% vs. 44.4%), and remained independently associated with OS and PFS (hazard ratio 2.55 and 2.24 per standard deviation), though discrimination was not significantly better than stage and residual disease alone. These biomarkers show subgroup-dependent prognostic value, and the CIRS provides an exploratory risk-stratification framework warranting prospective validation. Full article

(This article belongs to the Special Issue Molecular and Biomarker Advances in Gynecologic Oncology)

18 pages, 2785 KB

Open AccessArticle

Natriuretic Peptides as Predictors for the Diagnosis of Pulmonary Hypertension Secondary to Left Heart Disease and for the Assessment of Its Severity

by Filip Sawczak, Agata Kukfisz, Aleksandra Soloch, Kamila Kurkiewicz-Sawczak, Magdalena Dudek, Ewa Straburzyńska-Migaj and Marta Kałużna-Oleksy

Int. J. Mol. Sci. 2026, 27(17), 7776; https://doi.org/10.3390/ijms27177776 (registering DOI) - 30 Aug 2026

Abstract

Pulmonary hypertension (PH) frequently complicates heart failure with reduced ejection fraction (HFrEF) and worsens prognosis. Right heart catheterization (RHC) remains the diagnostic gold standard, but natriuretic peptides may help identify patients requiring invasive assessment. We retrospectively analyzed 563 HFrEF patients who underwent RHC. [...] Read more.

Pulmonary hypertension (PH) frequently complicates heart failure with reduced ejection fraction (HFrEF) and worsens prognosis. Right heart catheterization (RHC) remains the diagnostic gold standard, but natriuretic peptides may help identify patients requiring invasive assessment. We retrospectively analyzed 563 HFrEF patients who underwent RHC. Patients with and without PH were compared. Associations between B-type natriuretic peptide (BNP), N-terminal pro-B-type natriuretic peptide (NT-proBNP) and RHC parameters were assessed using correlations, restricted cubic splines, logistic regression and receiver operating characteristic (ROC) curves. PH was present in 443 patients (78.7%). Of the study group, 87 (15.5%) were females and 476 (84.5%) were males, the median age was 55 years and the median ejection fraction was 20%. Patients with PH had significantly higher BNP (p < 0.001), NT-proBNP (p < 0.001), New York Heart Association (NYHA) class (p < 0.001) and lower ejection fraction (p < 0.001). Increase in PH risk (p < 0.001), mean pulmonary artery pressure (p < 0.001), pulmonary vascular resistance (PVR) (p < 0.001) and pulmonary arterial wedge pressure (p < 0.001) and decrease in cardiac index (p < 0.001) were associated with an increase in BNP up to approximately 500 pg/mL and NT-proBNP up to approximately 3000 pg/mL. BNP and NT-proBNP were independent predictors of PH and PVR > 5 Wood units. BNP and NT-proBNP predicted PH with area under the curve (AUC) 0.798 and 0.727, respectively, while prediction of PVR > 5 Wood units (AUC 0.661 and 0.664, respectively) or cardiac index < 2.0 L/min/m² was less accurate (AUC 0.607 and 0.623, respectively). Measurement of plasma natriuretic peptides may support PH screening in HFrEF, but their limited ability to detect severe precapillary component confirms RHC as the definitive tool for hemodynamic phenotyping. Full article

(This article belongs to the Special Issue Molecular Pathology and Treatment of Heart Failure)

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22 pages, 31600 KB

Open AccessArticle

Chitosan Oligosaccharides Modulate Macrophage Inflammatory Signaling: Molecular Docking and Transcriptomic Evidence

by Yujun Sung, Siriporn Namwongsa, Sineenart Songkoomkrong, Supawadee Duangprom and Napamanee Kornthong

Int. J. Mol. Sci. 2026, 27(17), 7775; https://doi.org/10.3390/ijms27177775 (registering DOI) - 30 Aug 2026

Abstract

Oxidative stress-induced macrophage activation and vascular injury are major contributors to atherosclerosis, a chronic inflammatory disease associated with approximately 20 million deaths worldwide. This study investigated the antioxidant and anti-inflammatory activities of structurally characterized low-molecular-weight chitosan oligosaccharides (COS) derived from mud crab ( [...] Read more.

Oxidative stress-induced macrophage activation and vascular injury are major contributors to atherosclerosis, a chronic inflammatory disease associated with approximately 20 million deaths worldwide. This study investigated the antioxidant and anti-inflammatory activities of structurally characterized low-molecular-weight chitosan oligosaccharides (COS) derived from mud crab (Scylla olivacea) shell waste by hydrochloric acid hydrolysis and explored their molecular interactions with inflammation-related targets. Structural characterization by ¹³C-NMR and MALDI-TOF confirmed the identity of COS, while DPPH and ABTS analysis demonstrated concentration-dependent antioxidant activity. In LPS-induced RAW 264.7 macrophages, COS showed no cytotoxicity and significantly reduced nitric oxide production at 80 and 160 µg/mL. Molecular docking predicted favorable interactions of COS with several inflammation-associated proteins, with iNOS and COX-2 exhibiting the most favorable docking scores and extensive hydrogen-bonding and polar interactions. Transcriptomic profiling further revealed broad transcriptional remodeling and enrichment of pathways related to inflammation and atherosclerosis, including TNF, NF-κB, MAPK, Toll-like receptor, and lipid-and-atherosclerosis signaling. COS markedly suppressed inflammatory mediators, particularly Nos2 (iNOS) and Ptgs2 (COX-2) together with multiple cytokines and chemokines, consistent with reduced nitric oxide production and modulation of macrophage activation and foam cell-associated processes. Integration of docking and transcriptomic analyses identified iNOS and COX-2 as convergent candidate anti-inflammatory targets of COS, supporting its ability to attenuate inflammatory signaling through relevant pathways. These findings suggest that COS may modulate macrophage inflammatory responses through coordinated regulation of oxidative stress and inflammation-related signaling pathways. Full article

(This article belongs to the Special Issue Natural Products: Potential Anti-Inflammatory or Anti-Oxidative Mechanisms)

16 pages, 2049 KB

Open AccessReview

Targeting NR4A1 (Nur77) in Macrophage Polarization: Molecular Mechanisms and Translational Prospects for Cardiac Repair After Myocardial Infarction

by Jiaqian Lin, Jianmin Wang, Lei Sun and Rui Zhang

Int. J. Mol. Sci. 2026, 27(17), 7774; https://doi.org/10.3390/ijms27177774 (registering DOI) - 30 Aug 2026

Abstract

Sterile inflammation in the course of myocardial infarction can be a factor contributing to cardiac remodeling and heart failure. To break this pathological vicious circle, the polarization of macrophages to a reparative M2 phenotype, to start the tissue repair process and alleviate the [...] Read more.

Sterile inflammation in the course of myocardial infarction can be a factor contributing to cardiac remodeling and heart failure. To break this pathological vicious circle, the polarization of macrophages to a reparative M2 phenotype, to start the tissue repair process and alleviate the inflammation, is the key. Recent studies have shown that the nuclear receptor subfamily 4 group A (NR4A) orphan nuclear receptor family, particularly nuclear receptor subfamily 4 group A member 1 (NR4A1/NUR77), plays a role in macrophage reprogramming in relation to the modulation of macrophage polarization. Herein, we discuss in detail the specific pathways regulated by NR4A receptors during cellular reprogramming, including the nuclear factor kappa B (NF-κB) pathway and the oxidative phosphorylation pathway. We list below the beneficial effects of NR4A-treated macrophages in the hypoxic cardiac tissue, including their essential contribution to the formation of a neovascular network, the maintenance of the extracellular matrix by fibroblasts, and the direct rescue of remaining cardiomyocytes. In addition, given the translational potential of targeted therapy with immunomodulatory molecules against NR4A, we also review the design of macrophage-specific nanodelivery systems and the creation of novel small-molecule activators for ischemic cardiovascular diseases to identify therapeutic targets for cardiac repair based on NR4A. Full article

(This article belongs to the Section Molecular Immunology)

17 pages, 5511 KB

Open AccessArticle

Evolutionary Adaptability Coupled with Computation-Driven Engineering for Thermostability Enhancement of a Deoxynivalenol-Detoxifying Fusion Enzyme

by Yiting Pan, Hao Zhu, Qingwei Jiang, Bin Ma, Changhe Chen, Fengxia Lu, Huibing Chi and Ping Zhu

Int. J. Mol. Sci. 2026, 27(17), 7773; https://doi.org/10.3390/ijms27177773 (registering DOI) - 30 Aug 2026

Abstract

Deoxynivalenol (DON), a trichothecene mycotoxin commonly found in cereal grains and their derived products, poses significant risks to human and animal health. In previous work, a fusion enzyme composed of the dehydrogenase DADH and the aldo-keto reductase AKR13B3 was engineered to convert DON [...] Read more.

Deoxynivalenol (DON), a trichothecene mycotoxin commonly found in cereal grains and their derived products, poses significant risks to human and animal health. In previous work, a fusion enzyme composed of the dehydrogenase DADH and the aldo-keto reductase AKR13B3 was engineered to convert DON into the non-toxic 3-epi-DON in a single step. However, the poor thermal stability of this fusion enzyme limited its industrial application. In this study, EVcouplings and the GRAPE-WEB platform were utilized to identify key amino acid residues governing the thermal stability of the fusion enzyme AKR13B3–DADH. Through single-point mutation screening and the combination of beneficial mutation sites, a triple mutant M361L/T508Y/Y603F (M1) was obtained. The half-life of M1 at 50 °C reached about 500 min, representing a 16.4-fold increase compared with the wild type, while its catalytic activity increased by 2.7-fold. The apparent melting temperature increased by approximately 4 °C. Molecular dynamics simulations verified that the improved thermostability results from reduced conformational flexibility in key regions, enhanced structural packing, and a strengthened hydrogen bond network. These results demonstrate the successful development of a thermostable DON-detoxifying fusion enzyme and provide a practical basis for its industrial application. Full article

(This article belongs to the Section Biochemistry)

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32 pages, 2488 KB

Open AccessArticle

Low-Molecular-Weight Chitosan Produced by Gamma Irradiation and Hydrogen Peroxide Promotes Immune Readiness, Antioxidant Responses, and Survival in Photobacterium damselae-Challenged Asian Seabass (Lates calcarifer)

by Thitirat Rattanawongwiboon, Natthapong Paankhao, Ratchanon Chumchuen, Nattaset Chinnabutra, Wasin Saisin, Benchawan Kumwan, Pakapon Meachasompop, Yosapon Adisornprasert, Pimrawee Chaemlek, Prapansak Srisapoome, Passakorn Kingwascharapong, Theeranan Tangthong, Wararut Buncharoen and Anurak Uchuwittayakul

Int. J. Mol. Sci. 2026, 27(17), 7772; https://doi.org/10.3390/ijms27177772 (registering DOI) - 30 Aug 2026

Abstract

Low-molecular-weight chitosan may exhibit improved dispersion and biological accessibility compared with high-molecular-weight chitosan, but direct comparisons in Asian seabass remain limited. This study evaluated dietary low-molecular-weight chitosan produced by combined gamma irradiation and hydrogen peroxide depolymerization. The untreated and degraded preparations had apparent [...] Read more.

Low-molecular-weight chitosan may exhibit improved dispersion and biological accessibility compared with high-molecular-weight chitosan, but direct comparisons in Asian seabass remain limited. This study evaluated dietary low-molecular-weight chitosan produced by combined gamma irradiation and hydrogen peroxide depolymerization. The untreated and degraded preparations had apparent GPC-derived molecular weights of approximately 85 and 10 kDa, respectively. Juvenile Asian seabass (Lates calcarifer) were fed a Control diet of 1.0% high-molecular-weight chitosan or low-molecular-weight chitosan at 0.25%, 0.5%, or 1.0% for four weeks. Growth, serum antioxidant and humoral immune parameters, tissue-specific gene expression, and resistance to Photobacterium damselae were evaluated. No treatment significantly affected growth or feed utilization. Both high- and low-molecular-weight chitosan reduced serum MDA relative to the Control, whereas selected antioxidant and immune responses differed among inclusion levels. The 0.5% low-molecular-weight treatment increased GSH and GPx activity, the 1.0% low-molecular-weight treatment produced the highest CAT activity, and the 0.25% low-molecular-weight treatment produced the highest total serum IgM. Low-molecular-weight chitosan generally produced broader tissue-specific transcriptional responses than 1.0% high-molecular-weight chitosan. Following bacterial challenge, all chitosan treatments improved survival relative to the Control, with the highest numerical RPS observed in the 0.5% low-molecular-weight group. These results indicate that molecular-weight reduction influenced the biological response to dietary chitosan. Among the tested levels, 0.5% produced the most consistent combined response, although the optimal level was endpoint-specific and requires longer-term validation. Full article

(This article belongs to the Special Issue IJMS 25th Anniversary: Updates and Progress on Bioactives and Nutraceuticals)

16 pages, 4389 KB

Open AccessArticle

Annexin A5 Maintains Mitochondrial Integrity by Inhibiting mPTP Opening to Protect Against Acetaminophen-Induced Acute Liver Injury

by Xiaowen Zhang, Wenwei Li, Luqi Li, Ying Wang, Wei Tang, Jing Zhang and Zichun Hua

Int. J. Mol. Sci. 2026, 27(17), 7771; https://doi.org/10.3390/ijms27177771 (registering DOI) - 30 Aug 2026

Abstract

Acetaminophen (APAP) represents a major cause of drug-induced liver injury (DILI), and effective pharmacological interventions remain limited. Annexin A5 (AnxA5), a Ca²⁺-dependent phospholipid-binding protein, participates in diverse biological processes related to tissue repair. In this study, we evaluated AnxA5 expression in [...] Read more.

Acetaminophen (APAP) represents a major cause of drug-induced liver injury (DILI), and effective pharmacological interventions remain limited. Annexin A5 (AnxA5), a Ca²⁺-dependent phospholipid-binding protein, participates in diverse biological processes related to tissue repair. In this study, we evaluated AnxA5 expression in APAP-challenged mouse livers and clinical samples from patients with liver injury. Using hepatic cell lines AML12 and HepG2, we performed overexpression-based functional assays to assess the cytoprotective effects of AnxA5 against APAP toxicity. Co-immunoprecipitation assays were applied to characterize protein interactions, and mitochondrial functional parameters were measured to dissect the underlying molecular mechanism. We found that AnxA5 was significantly upregulated in both APAP exposed mice and APAP DILI patients. Cellular functional assays showed that AnxA5 overexpression mitigated APAP triggered cytotoxicity in AML12 and HepG2 cells. Mechanistically, AnxA5 bound to voltage dependent anion channel 1 (VDAC1), restrained VDAC1 mediated mitochondrial Ca²⁺ influx, and suppressed VDAC1 oligomerization, which further inhibited mitochondrial permeability transition pore (mPTP) opening. In an APAP-induced liver injury mouse model, exogenous recombinant AnxA5 treatment maintained mitochondrial integrity and ameliorated hepatic inflammation and liver damage. Collectively, our data reveal AnxA5 as an endogenous mitochondrial protective factor and support its therapeutic potential against APAP-induced liver injury. Full article

(This article belongs to the Section Biochemistry)

12 pages, 2608 KB

Open AccessArticle

Nanoscale Electromechanical and Conductive Properties of a Layered Two-Dimensional Hybrid Perovskite

by Hee-Chang Jeon, Woohyuk Jang, Jiseon Yun, Sein Min, Joong Yeon Lim and Young-Seong Kim

Int. J. Mol. Sci. 2026, 27(17), 7770; https://doi.org/10.3390/ijms27177770 (registering DOI) - 30 Aug 2026

Abstract

Two-dimensional (2D) organic–inorganic hybrid perovskites exhibit coupled ionic, electronic, and electromechanical responses that can strongly influence local charge transport. Here, solution-processed mixed-halide butylammonium lead perovskite crystals were mechanically exfoliated and investigated using X-ray diffraction, atomic force microscopy, piezoresponse force microscopy (PFM), and conductive [...] Read more.

Two-dimensional (2D) organic–inorganic hybrid perovskites exhibit coupled ionic, electronic, and electromechanical responses that can strongly influence local charge transport. Here, solution-processed mixed-halide butylammonium lead perovskite crystals were mechanically exfoliated and investigated using X-ray diffraction, atomic force microscopy, piezoresponse force microscopy (PFM), and conductive atomic force microscopy (c-AFM). PFM measurements under −5, 0, and +5 V revealed clear bias-dependent changes in amplitude and phase, indicating an electric field-sensitive local electromechanical response. Local c-AFM measurements showed nonlinear bipolar hysteresis, with a pronounced increase in current near +7–8 V and a decrease near −7 to −6 V during the subsequent negative sweep. Because the crystals are mixed ionic–electronic conductors and the nanoscale tip–sample junction introduces substantial injection and contact barriers, the observed behavior is interpreted as resistive switching-like conductivity modulation, rather than definitive ferroelectric switching. The results are consistent with the combined contributions of charge injection, trap filling, possible ionic redistribution, and piezoelectricity-associated modulation of the local transport barrier. These findings provide nanoscale insight into electric field-dependent electromechanical and out-of-plane conductive behaviors in layered 2D hybrid perovskites. Full article

(This article belongs to the Special Issue Exploring Molecular-Scale and Nano-Scale Materials: Design, Characterization and Application)

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13 pages, 888 KB

Open AccessArticle

Proteomic Profile Differences in Immune-Related Diseases in Pediatric Patients Under Five Years Old: Asthma and IgE-Dependent Allergies—A Pilot Study

by Natalia Rzetecka-Mańka, Joanna Matysiak, Eliza Matuszewska-Mach, Paulina Sobkowiak, Irena Wojsyk-Banaszak, Anna Bręborowicz, Paulina Borysewicz, Agnieszka Klupczyńska-Gabryszak and Jan Matysiak

Int. J. Mol. Sci. 2026, 27(17), 7769; https://doi.org/10.3390/ijms27177769 (registering DOI) - 30 Aug 2026

Abstract

Asthma is a heterogeneous disease that often begins in childhood and frequently occurs alongside allergic conditions. In asthma research, it is important to focus on proteins that are the primary regulators of cellular physiology. The differences in the proteome between children with asthma [...] Read more.

Asthma is a heterogeneous disease that often begins in childhood and frequently occurs alongside allergic conditions. In asthma research, it is important to focus on proteins that are the primary regulators of cellular physiology. The differences in the proteome between children with asthma and those with an atopic background remain poorly understood. The present study included 130 serum samples from four groups of pediatric patients under the age of five: (1) with asthma and IgE-dependent allergies; (2) with non-atopic asthma; (3) non-asthmatics with IgE-dependent allergy; and (4) a control group without asthma and IgE-dependent allergies. The serum samples were used for protein–peptide profiling and proteomic identification using nanoLC-MALDI-TOF/TOF MS/MS. The obtained data were analyzed using univariate statistics and the STRING tool v12.0 to identify protein–protein potential interactions. A total of seven proteins were identified as discriminative between the study groups: A2M, AACT, IgG3, C3, ITIH2, IgG3 and IGK. All of them were upregulated in patients with IgE-dependent allergy compared to other study groups. STRING analysis identified functional associations among four proteins (AACT, A2M, C3, and ITIH2) with discriminatory potential for distinguishing between non-atopic asthma and non-asthmatic patients with IgE-dependent allergy. The results suggest that the identified putative protein markers overlap in cellular pathways, including those associated with the pathophysiology of asthma and allergic disorders. These findings provide further insight into the overall proteomic profile of pediatric patients with asthma and IgE-dependent allergy, highlighting its heterogeneity across the analyzed groups. Full article

(This article belongs to the Special Issue Molecular Research in Asthma and Allergy)

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24 pages, 5261 KB

Open AccessArticle

Pullulan Fatty Ester Nanocoatings with Tunable Surface Wettability and Bio-Inert Surface Properties

by Femke De Ceulaer, Hao-Chun Chiu, Olivier Deschaume, Carmen Bartic and Pedro Fardim

Int. J. Mol. Sci. 2026, 27(17), 7768; https://doi.org/10.3390/ijms27177768 (registering DOI) - 30 Aug 2026

Abstract

Polysaccharide derivatives with tunable hydrophobicity offer a versatile platform for tailoring material properties through controlled chemical modification. In this study, pullulan was functionalized with saturated fatty acids (C14–C18) via N,N-carbonyldiimidazole-mediated esterification, yielding degrees of substitution between 0.24 and 0.77. The effect of alkyl [...] Read more.

Polysaccharide derivatives with tunable hydrophobicity offer a versatile platform for tailoring material properties through controlled chemical modification. In this study, pullulan was functionalized with saturated fatty acids (C14–C18) via N,N-carbonyldiimidazole-mediated esterification, yielding degrees of substitution between 0.24 and 0.77. The effect of alkyl chain length and substitution levels on the thermal, structural, and surface properties of the resulting pullulan esters was systematically investigated. Increasing chain length enhanced thermal stability and promoted structural ordering, with stearate-modified pullulan exhibiting melting transitions at 40–42 °C and higher degradation temperatures (≥312 °C). Spin-coated nanoscale coatings (110–150 nm thick) exhibited extremely smooth surfaces (S_a < 1 nm), although differences in solubility influenced solvent evaporation and induced nanostructural variations. Surface wettability increased systematically with alkyl chain length and degree of substitution, resulting in water contact angles between 85° and 105°. All coatings were non-cytotoxic and inhibited bacterial growth at the interface without leaching, suggesting a contact-dependent antibacterial effect. These results establish clear structure–property relationships in pullulan esters, demonstrating that surface wettability, structural ordering, and surface functionality can be systematically adjusted via alkyl chain length and degree of substitution. Full article

(This article belongs to the Special Issue Advanced Functional Materials and Their Applications in Energy, Environment and Biomedical Fields)

22 pages, 7616 KB

Open AccessArticle

Individual and Cooperative Photochemical–Enzymatic Processes for the Degradation of the Dye Bromothymol Blue: Kinetic and Eco-Toxicity Analysis

by Andrea S. Urquiza, Agustina Reynoso, M. Alicia Biasutti, Hernán A. Montejano and Eugenia Reynoso

Int. J. Mol. Sci. 2026, 27(17), 7767; https://doi.org/10.3390/ijms27177767 (registering DOI) - 30 Aug 2026

Abstract

The degradation of bromothymol blue (BTB) in aqueous solution was investigated through individual and sequential photochemical and enzymatic treatments. Photodegradation experiments were performed under UVC, UVB, UVA and visible irradiation at different pH values and atmosphere conditions, while enzymatic degradation was evaluated using [...] Read more.

The degradation of bromothymol blue (BTB) in aqueous solution was investigated through individual and sequential photochemical and enzymatic treatments. Photodegradation experiments were performed under UVC, UVB, UVA and visible irradiation at different pH values and atmosphere conditions, while enzymatic degradation was evaluated using Laccase from Trametes versicolor under varying pH, temperature and enzyme concentration. Kinetic analyses were performed in all cases. BTB degradation was strongly dependent on irradiation wavelength and pH. The highest photodegradation rates were obtained under UVC irradiation, particularly in alkaline medium. Additionally, our results suggest that BTB photolysis mainly proceeds through a unimolecular pathway. On the other hand, enzymatic degradation was favored at acidic pH, elevated temperature, and high amount of laccase, with optimal performance observed at pH 5 and 40 °C. Sequential treatments combining photochemical and enzymatic processes improved the overall removal efficiency, reaching degradation values above 70% regardless of the treatments order. Ecotoxicological evaluation using the Vibrio fischeri bioluminescence inhibition assay revealed a significant reduction in toxicity after all treatments, particularly those involving UVC irradiation. These results demonstrate the complementary nature of both processes and highlight the potential of combined photochemical–enzymatic treatments for the remediation of dye-contaminated waters. Full article

(This article belongs to the Special Issue Photophysics and Photochemistry in Biological Molecules)

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27 pages, 4430 KB

Open AccessReview

Molecular Mechanisms of Endocrine-Disrupting Chemicals and Emerging-Pollutant Toxicity in Human Reproduction: From Xenobiotic Exposure to Fertility Impairment and Reproductive Carcinogenesis

by Zakhia El Beaino, Jean-Marc Ayoubi and Samir Hamamah

Int. J. Mol. Sci. 2026, 27(17), 7766; https://doi.org/10.3390/ijms27177766 (registering DOI) - 30 Aug 2026

Abstract

Human fertility is declining across industrialised populations, while the incidence of hormone-dependent reproductive cancers rises. Endocrine-disrupting chemicals (EDCs) and structurally related emerging pollutants are implicated in both. These two outcomes are generally reviewed as separate studies. This review argues that they are two [...] Read more.

Human fertility is declining across industrialised populations, while the incidence of hormone-dependent reproductive cancers rises. Endocrine-disrupting chemicals (EDCs) and structurally related emerging pollutants are implicated in both. These two outcomes are generally reviewed as separate studies. This review argues that they are two latencies of a single molecular toxicology. The compounds concerned are structurally diverse: phthalates, bisphenols, per- and polyfluoroalkyl substances (PFASs), pesticides, polychlorinated biphenyls (PCBs) and dioxins, brominated and organophosphate flame retardants, pharmaceuticals and personal-care products (PPCPs), and micro- and nanoplastics. They nonetheless converge on a limited repertoire of molecular lesions. These include the disruption of hypothalamic–pituitary–gonadal (HPG) signalling through kisspeptin/GnRH and gonadotropin gene expression and interference at nuclear and membrane hormone receptors (ERα/β, AR, GPER, thyroid receptors, AhR, PPARγ). They also include the inhibition of steroidogenesis at StAR and the CYP11A1–CYP17A1–CYP19A1/3β-HSD/17β-HSD cascade and reactive-oxygen-species generation with mitochondrial dysfunction and Keap1–Nrf2 disruption. Epigenetic reprogramming through DNA methylation, histone modification and non-coding RNAs, together with crosstalk with metabolic and immune signalling, completes the set. These lesions produce measurable cytotoxic and genotoxic damage to gametes and the early embryo: sperm DNA fragmentation and 8-oxo-dG accumulation, blood–testis-barrier breakdown, oocyte meiotic-spindle defects, and granulosa-cell apoptosis and pyroptosis. The same receptor, oxidative and genotoxic hubs drive hormone-dependent reproductive carcinogenesis over longer latencies. The review makes three contributions. First, it traces these shared hubs continuously from fertility impairment to malignancy rather than treating them as separate fields. Second, it grades the certainty of the human evidence class by class, so that robust associations can be distinguished from provisional ones. Third, it integrates pseudo-persistent pollutants alongside the classical persistent compounds. These are micro- and nanoplastics, which act as both toxicants and vectors for adsorbed co-contaminants, and pharmaceutical and personal-care residues sustained by continuous wastewater input. Their inclusion demonstrates that chronic low-dose exposure does not require chemical persistence. We conclude with mitigation strategies and an explicit account of what the current evidence base cannot yet support. Full article

(This article belongs to the Special Issue Toxicity Mechanism of Emerging Pollutants: 2nd Edition)

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18 pages, 834 KB

Open AccessReview

Unique Features of Melanoma Risk and Diagnosis in Red-Haired Populations

by Kiran R. Ebrahimi, Stephen M. Ostrowski and David E. Fisher

Int. J. Mol. Sci. 2026, 27(17), 7765; https://doi.org/10.3390/ijms27177765 (registering DOI) - 30 Aug 2026

Abstract

Cutaneous melanoma is the most lethal skin cancer, and risk is strongly shaped by pigmentary phenotype. The red-hair color phenotype, marked by red hair, fair skin, freckling, and poor tanning, carries elevated melanoma risk driven largely by loss-of-function variants in the melanocortin 1 [...] Read more.

Cutaneous melanoma is the most lethal skin cancer, and risk is strongly shaped by pigmentary phenotype. The red-hair color phenotype, marked by red hair, fair skin, freckling, and poor tanning, carries elevated melanoma risk driven largely by loss-of-function variants in the melanocortin 1 receptor gene (MC1R). This risk is not explained by reduced ultraviolet protection alone. Impaired MC1R signaling shifts melanogenesis from photoprotective eumelanin toward pheomelanin, a pigment associated with oxidative stress and partly ultraviolet-independent melanomagenesis. MC1R may also influence melanoma susceptibility through pigment-independent effects on DNA damage responses, repair signaling, and genomic stability. These mechanisms support investigation of MC1R genotype, visible phenotype, nevus burden, pigment chemistry, and imaging-derived lesion metrics as complementary tools for risk stratification. Diagnosis is also distinctive in this population. Amelanotic and hypomelanotic melanomas are associated with the red-hair color phenotype, and their low pigmentary contrast may delay recognition and contribute to diagnosis at a more advanced stage. This review integrates genetic, molecular, biomarker, diagnostic, and therapeutic literature specific to red-haired populations. We argue that elevated biological susceptibility and diagnostic difficulty compound one another, and we outline priorities for MC1R-informed surveillance, imaging adapted to pigment-poor disease, pharmacologic modulation of MC1R-related pathways, and prospective risk models integrating genotype, phenotype, nevus burden, pigment biology, and imaging. Full article

(This article belongs to the Special Issue Skin Cancer: Molecular Drivers, Biomarkers, Non-Invasive Diagnostics and Therapeutic Strategies)

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21 pages, 3341 KB

Open AccessArticle

PMAVP: A Mamba-Deep Learning Framework for Antiviral Peptide Identification and Functional Activity Prediction

by Peiwei Wei, Weihao Su, Qingsong Qin, Chuliang Wei, Yi Shi and Guishan Zhang

Int. J. Mol. Sci. 2026, 27(17), 7764; https://doi.org/10.3390/ijms27177764 (registering DOI) - 30 Aug 2026

Abstract

Accurate computational prediction of antiviral peptides (AVPs) can accelerate peptide screening and reduce experimental costs. However, existing deep learning-based methods still suffer from severe class imbalance, over-reliance on handcrafted features and limited interpretability. Here, we propose PMAVP, a multi-task learning framework that integrates [...] Read more.

Accurate computational prediction of antiviral peptides (AVPs) can accelerate peptide screening and reduce experimental costs. However, existing deep learning-based methods still suffer from severe class imbalance, over-reliance on handcrafted features and limited interpretability. Here, we propose PMAVP, a multi-task learning framework that integrates the ProtT5 pre-trained protein language model with a Mamba-inspired module for AVP identification and functional activity prediction. We use ProtT5 to extract deep semantic representations from peptide sequences and a Mamba module to capture long-range dependencies at a lower computational complexity. We introduce Focal Loss to mitigate class imbalance and leverage transfer learning to enhance performance on functional activity prediction. Experimental results demonstrate that our model achieves superior performance in terms of prediction accuracy, stability, and computational efficiency. Furthermore, DeepSHAP-based interpretability analysis reveals that the first 40 amino acid residues contribute substantially to AVP prediction. Full article

(This article belongs to the Special Issue Machine Learning Applications in Bioinformatics and Biomedicine: 4th Edition)

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