International Journal of Molecular Sciences

20 pages, 7105 KB

Open AccessArticle

Dengue Virus NS5 Target Discovery: A Comprehensive in Silico Exploration of Novel Druggable Sites for Pan-Serotype Antiviral Design

by João Trigueiro-Louro, Vanessa Correia, Inara dos Santos Ali, Bulbul Ahmed and Helena Rebelo-de-Andrade

Int. J. Mol. Sci. 2026, 27(12), 5639; https://doi.org/10.3390/ijms27125639 - 22 Jun 2026

Viewed by 803

Abstract

Dengue is the most common vector-borne viral disease worldwide, posing an increasing global health threat. Despite its high burden, no approved antiviral treatments or widely applicable vaccines exist, and patient management remains limited to supportive care, underscoring the urgent need for antiviral development. [...] Read more.

Dengue is the most common vector-borne viral disease worldwide, posing an increasing global health threat. Despite its high burden, no approved antiviral treatments or widely applicable vaccines exist, and patient management remains limited to supportive care, underscoring the urgent need for antiviral development. The NS5 protein is a prime antiviral target, owing to its crucial role in viral replication, high conservation across dengue virus (DENV) serotypes and lack of a human orthologue. We conducted a comprehensive sequence-to-structure analysis to identify conserved druggable regions within NS5, integrating large-scale sequence analysis with structural characterization across all four DENV serotypes. We identified four highly promising Consensus Druggable Pockets within the NS5 dimer—CDP1d, CDP3d, CDP5d and CDP12d—that overlap functionally critical regions, alongside 149 new potential hot spot residues. Domain-specific analysis revealed that MTase offers more densely conserved targets, whereas RdRp provides broader druggable surfaces, revealing complementary features for pharmacological modulation. Several identified pockets spatially overlap known inhibitor binding sites, and preliminary docking analyses support their capacity to accommodate small molecules, reinforcing their therapeutic relevance as candidate targets. Collectively, these findings provide a robust framework for the rational design of pan-serotype anti-DENV NS5 antivirals with an enhanced barrier to resistance. Full article

(This article belongs to the Section Molecular Informatics)

► Show Figures

Figure 1

13 pages, 2789 KB

Open AccessArticle

Orthogroup-Based Comparative Analysis of Prophage Gene Content in Candidatus Liberibacter Asiaticus Supports a Predominantly Conserved Global Repertoire with Limited Accessory Variation

by Abdullah F. Alhashel, Ali A. Almasrahi, Mohammed A. Alsaleh, Arya Widyawan, Mahmoud H. El-Komy and Yasser E. Ibrahim

Int. J. Mol. Sci. 2026, 27(12), 5638; https://doi.org/10.3390/ijms27125638 - 22 Jun 2026

Viewed by 442

Abstract

Huanglongbing, a destructive citrus disease of global importance that is also present in Saudi Arabia, is associated with Candidatus Liberibacter asiaticus (CLas) and remains a major threat to citrus production. Although previous studies have documented sequence variation and prophage polymorphism in CLas, broader [...] Read more.

Huanglongbing, a destructive citrus disease of global importance that is also present in Saudi Arabia, is associated with Candidatus Liberibacter asiaticus (CLas) and remains a major threat to citrus production. Although previous studies have documented sequence variation and prophage polymorphism in CLas, broader comparisons of prophage-associated gene content remain limited. In particular, comparative orthogroup analysis of prophage gene-content conservation across geographically structured CLas populations has rarely been explored. In this study, we analyzed 42 CLas prophage genomes from Saudi Arabia and other geographic regions using a comparative orthogroup framework. OrthoFinder assigned 99.1% of predicted proteins (1825 of 1841) to 64 orthogroups, with only 16 genes remaining unassigned. A small number of rare orthogroups restricted to only a few genomes were identified, and no orthogroup was detected in all genomes. Presence–absence analyses supported a predominantly conserved prophage gene repertoire together with a small accessory component, while also indicating that apparent absences should be interpreted in light of mixed assembly status and prophage-region completeness. Saudi Arabian genomes were distributed within the broader global framework and exhibited generally similar gene-content profiles rather than a deeply separated lineage. Functional interpretation of representative orthogroups identified conserved prophage-associated genes related to replication, helicase activity, and phage packaging, whereas variable orthogroups were primarily associated with hypothetical or accessory prophage-related functions. Overall, these results are consistent with a model in which CLas prophage diversification is associated more with sequence-level variation and localized structural differences than with extensive gain or loss of prophage genes. These findings further refine current understanding of CLas genome evolution and highlight conserved prophage-associated targets that may support molecular diagnostics and epidemiological surveillance. Full article

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

► Show Figures

Figure 1

21 pages, 13573 KB

Open AccessArticle

Caveolin-1 Attenuates Excitotoxic Signaling by Regulating NMDA, AMPA, and Kainate Receptor-Mediated Calcium Influx in Hippocampal Neuronal Cultures

by Swapna Kannothum Kandy, Madhura Milind Nimonkar, Suravi Sasmita Dash, Prashanth N. Vashista, Bhupesh Mehta and Yogananda S. Markandeya

Int. J. Mol. Sci. 2026, 27(12), 5637; https://doi.org/10.3390/ijms27125637 - 22 Jun 2026

Viewed by 1608

Abstract

Glutamate excitotoxicity is a critical pathological mechanism underlying neuronal death in ischemic stroke, epilepsy, and neurodegenerative diseases. Caveolin-1 (Cav-1), a structural protein of caveolae membrane microdomains, has emerged as a potential modulator of neuronal survival, yet its precise mechanisms in excitotoxicity remain incompletely [...] Read more.

Glutamate excitotoxicity is a critical pathological mechanism underlying neuronal death in ischemic stroke, epilepsy, and neurodegenerative diseases. Caveolin-1 (Cav-1), a structural protein of caveolae membrane microdomains, has emerged as a potential modulator of neuronal survival, yet its precise mechanisms in excitotoxicity remain incompletely understood. In this study, we investigated the role of Cav-1 in regulating glutamate-induced calcium dysregulation, reactive oxygen species (ROS) generation, and mitochondrial dysfunction in primary hippocampal neurons. Using Cav-1 overexpression (Cav-1OE) and Cav-1 knockdown (Cav-1KD) approaches, we demonstrate that Cav-1OE significantly attenuates glutamate-stimulated intracellular Ca²⁺ elevation, reduces ROS generation, and prevents mitochondrial membrane potential (Ψ_m) depolarization. Further investigation revealed that Cav-1OE reduces, while Cav-1KD enhances, calcium responses mediated by NMDA, AMPA, and KA receptors. These findings establish that Cav-1 functionally attenuates excitotoxic signaling by negatively regulating ionotropic glutamate receptor-mediated Ca²⁺ influx. Full article

(This article belongs to the Special Issue Molecular Targets and Toxic Interactomes of Pathogenic Proteins in Neurodegeneration)

► Show Figures

Graphical abstract

14 pages, 518 KB

Open AccessReview

The Microbiota in the Diagnosis and Treatment of Autism Spectrum Disorder

by Ekaterina A. Trifonova, Roman A. Ivanov, Alex V. Kochetov and Sergey A. Lashin

Int. J. Mol. Sci. 2026, 27(12), 5636; https://doi.org/10.3390/ijms27125636 - 22 Jun 2026

Viewed by 536

Abstract

Autism spectrum disorder (ASD) is a complex neurodevelopmental condition characterized by impaired social interaction, communication deficits, and repetitive behaviors. Recent research highlights the role of the gut microbiota in ASD pathophysiology, particularly through the microbiota–gut–brain axis. The microbiota may influence neurodevelopment via multiple [...] Read more.

Autism spectrum disorder (ASD) is a complex neurodevelopmental condition characterized by impaired social interaction, communication deficits, and repetitive behaviors. Recent research highlights the role of the gut microbiota in ASD pathophysiology, particularly through the microbiota–gut–brain axis. The microbiota may influence neurodevelopment via multiple signaling pathways, including the GABAergic and serotonergic systems, as well as the mTOR signaling pathway. This analytical review examines current evidence on microbiota alterations in ASD and evaluates microbiota-targeted strategies for diagnosis and treatment, focusing on fecal microbiota transplantation (FMT), probiotics, and diet-based therapeutic approaches. The review also provides a critical appraisal of the microbiota–gut–brain axis in the context of autism spectrum disorder. Full article

(This article belongs to the Special Issue Molecular Research of Gut Microbiota in Human Health and Diseases)

► Show Figures

Figure 1

23 pages, 12189 KB

Open AccessArticle

Activated Macrophages Promote TNF-α-Associated Tumor Cell Necroptosis in Pituitary Apoplexy Through the PIEZO1–NFATC2/REL Axis

by Xingbo Li, Luowen Zhou, Zhuowei Lei, Sihan Li, Quanji Wang, Haochen Zhao, Linpeng Xu, Juan Chen, Xueyan Wan, Yimin Huang and Ting Lei

Int. J. Mol. Sci. 2026, 27(12), 5635; https://doi.org/10.3390/ijms27125635 - 22 Jun 2026

Viewed by 566

Abstract

Pituitary apoplexy is an uncommon but clinically urgent complication that often involves intrasellar hemorrhage and tissue necrosis. The mechanisms linking acute tissue injury to the inflammatory tumor microenvironment remain incompletely defined. Here, we characterized the apoplexy-associated microenvironment and examined whether macrophage mechanosensitive signaling [...] Read more.

Pituitary apoplexy is an uncommon but clinically urgent complication that often involves intrasellar hemorrhage and tissue necrosis. The mechanisms linking acute tissue injury to the inflammatory tumor microenvironment remain incompletely defined. Here, we characterized the apoplexy-associated microenvironment and examined whether macrophage mechanosensitive signaling contributes to inflammatory amplification and tissue damage in pituitary neuroendocrine tumors (PitNETs). We combined single-cell RNA sequencing (scRNA-seq), histological validation, clinical stratification, and in vitro functional assays using apoplectic and non-apoplectic human PitNET specimens. Macrophage state transitions, intercellular communication, and transcriptional regulatory programs were analyzed, followed by an experimental assessment of the PIEZO1–Ca²⁺ axis and macrophage-conditioned medium-induced tumor cell death. Histological validation confirmed macrophage accumulation in apoplectic PitNETs, including a 1.67-fold increase in IBA-1-positive cells (p < 0.001). CellChat-inferred interaction metrics increased descriptively in apoplectic samples. Apoplectic tissues showed higher TNF-α expression (3.00-fold; p < 0.0001) and higher PIEZO1 fluorescence in IBA-1-positive regions (1.39-fold; p = 0.001). Yoda1 increased Calcium 520 fluorescence in macrophages (1.72-fold; p = 0.002), whereas Piezo1 knockdown reduced the Yoda1-associated response (p = 0.003). Conditioned medium from activated macrophages increased total Annexin V/PI-positive death in AtT-20 cells (0.53 ± 0.53% to 32.48 ± 1.14%; p < 0.001) and GH3 cells (0.82 ± 0.50% to 30.92 ± 1.11%; p < 0.001); Piezo1 knockdown or TNF-α neutralization attenuated this effect. Clinically, pathological necrosis was associated with higher symptom frequencies and a greater adjusted likelihood of two or more clinical symptoms. Together, these findings indicate that PIEZO1-related macrophage signaling may participate in TNF-α-associated tumor cell necroptosis in pituitary apoplexy. Pathological necrosis was linked to greater acute symptom burden and perioperative hormonal abnormalities, suggesting that it may identify a clinically severe apoplexy subtype. Full article

(This article belongs to the Section Molecular Immunology)

► Show Figures

Graphical abstract

21 pages, 18036 KB

Open AccessArticle

Localization and Biological Activities of Bioflavonoids from Taxus canadensis Marshall

by Svetlana M. Zaytseva, Elena A. Kalasnikova, Rima N. Kirakosyan, Jing Liang, Elizaveta A Bolotina and Nikolay A. Trusov

Int. J. Mol. Sci. 2026, 27(12), 5634; https://doi.org/10.3390/ijms27125634 - 22 Jun 2026

Cited by 1 | Viewed by 475

Abstract

Relict yew plants (Taxus L.) are not only ornamental plants with valuable wood but also have the ability to synthesize the unique compound taxol, which is successfully used in the treatment of cancer due to its powerful cytotoxic effect. Due to the [...] Read more.

Relict yew plants (Taxus L.) are not only ornamental plants with valuable wood but also have the ability to synthesize the unique compound taxol, which is successfully used in the treatment of cancer due to its powerful cytotoxic effect. Due to the presence of taxol, all parts of yew plants are extremely poisonous, but there have been cases where animals have eaten yew cones without fatal consequences. The biosynthesis of taxol is carried out due to the interaction of the isoprenoid and phenolic pathways of the secondary metabolism of plants. Despite the close attention of researchers to the peculiarities of taxol metabolism, there is very little data on the tissue and intracellular localization of both taxols and phenolic compounds in yew plants. Polyphenols are known to be physiologically active mediators involved in respiration, photosynthesis, plant growth and development, as well as in the process of in vitro dedifferentiation. Since Taxus is a relict species and has a limited and hard-to-reach range in nature, technologies that allow yew plants to be restored without removing plant material from the natural environment are of great practical importance: overcoming deep physiological dormancy of seeds, microclonal reproduction and initiation of plant growth. In vitro cultures are possible sources of biologically active and medicinal products. The aims and objectives of this study are to determine the characteristics of the formation and localization of phenolic compounds with high biological activity in various organs of plants of the genus Taxus and to determine the biological activity of ethanolic extracts from this plant. The objects of this study were the generative organs of Taxus canadensis, collected during the entire growing season (April–October) from plants growing in the Moscow region. The localization of various classes of polyphenols was determined by histochemical methods using light microscopy. Histochemical studies have shown the abundant presence of polyphenols in yew megastrobiles, microstrobiles, cones, seeds and aril. Ethanolic plant extracts were used to determine the biological activity. Flavans were dominant in the aril at various stages of vegetation, which was confirmed by our biochemical and histochemical studies. Extractive substances of T. canadensis show high antibacterial activity, especially in its shoot extracts. Ethanolic extracts from plant shoots showed greater biological activity than seed extracts. Aril extracts had the lowest cytotoxicity. Full article

(This article belongs to the Special Issue Extraction and Application of Natural Compound)

► Show Figures

Figure 1

19 pages, 758 KB

Open AccessArticle

Systemic Molecular Alterations of TP53, SIRT-1, and miR-34a Expression in Atrial Fibrillation: A Prospective Exploratory Biomarker Study

by Monika Różycka-Kosmalska, Izabela Szymczak-Pajor, Agnieszka Śliwińska, Małgorzata Kozłowska, Jerzy Krzysztof Wranicz and Marcin Kosmalski

Int. J. Mol. Sci. 2026, 27(12), 5633; https://doi.org/10.3390/ijms27125633 - 22 Jun 2026

Viewed by 606

Abstract

p53, miR-34a, and SIRT-1 are involved in cellular stress responses, senescence, and inflammation—processes central to the pathophysiology of atrial fibrillation (AF). In this study, circulating TP53 and SIRT-1 serum miR-34a expression were determined in patients with and without AF, in order to assess [...] Read more.

p53, miR-34a, and SIRT-1 are involved in cellular stress responses, senescence, and inflammation—processes central to the pathophysiology of atrial fibrillation (AF). In this study, circulating TP53 and SIRT-1 serum miR-34a expression were determined in patients with and without AF, in order to assess their associations with AF. We also checked their potential diagnostic utility as systemic biomarkers associated with AF. The study included 189 adults, 94 AF+, 95 AF−. Clinical, anthropometric, and biochemical data were collected. Whole-blood TP53 and SIRT-1 mRNA expression and serum miR-34a expression were quantified by RT-qPCR. ROC analysis and Youden-derived odds ratios assessed exploratory diagnostic performance. AF patients had significantly higher expression of TP53 (0.0352 vs. 0.0253; p < 0.001) and miR-34a (0.0215 vs. 0.0099; p < 0.001), but significantly lower expression of SIRT-1 (0.0079 vs. 0.0145; p < 0.001). The level of SIRT-1 expression showed the highest discriminatory performance (exploratory AUC = 0.6987; p < 0.0001). TP53 expression levels exceeding 0.0295 were associated with nearly threefold higher odds of AF (OR = 2.92, 95% CI: 1.61–5.28, p = 0.0006), whereas the expression levels of SIRT-1 and miR-34a were not significantly associated with AF in cut-off analysis. In the AF group, a positive correlation was found between the expression of TP53 and SIRT-1 (Rho = 0.3609, p < 0.001); however, it was not consistent with a canonical model of miR-34a-mediated SIRT-1 suppression. In turn, the expression of miR-34a correlated positively with age and C-reactive protein level and negatively with estimated glomerular filtration rate (eGFR). The obtained results suggest that AF is associated with altered expression of circulating TP53, SIRT-1, and miR-34a. However, due to the fact that the expression levels were measured in peripheral compartments, and not in atrial tissue, the obtained results should not be interpreted as direct evidence of AF-related atrial remodeling. For these reasons, further investigations involving simultaneous measurements of the TP53/miR-34a/SIRT-1 regulatory axis, both in the circulating compartment and atrial tissue, should be performed. Full article

(This article belongs to the Special Issue Molecular and Translational Mechanisms of Cardiac Arrhythmias: From Oxidative Stress to Novel Therapeutic Strategies)

► Show Figures

Figure 1

44 pages, 3073 KB

Open AccessReview

From Chronic Inflammation to Malignancy: Molecular Mechanisms and Therapeutic Insights in Oral Carcinogenesis

by Ying-Jia Huang, Gaiping Shi, Fengyuan Lv, Ronghua Deng, Qingfeng Zhan, Zixuan Zhang, Jiangyuan Song and Zhi Xu

Int. J. Mol. Sci. 2026, 27(12), 5632; https://doi.org/10.3390/ijms27125632 - 22 Jun 2026

Cited by 1 | Viewed by 1175

Abstract

Oral squamous cell carcinoma (OSCC) frequently develops within chronically injured oral mucosa and may be preceded by clinically recognizable oral potentially malignant disorders (OPMDs), which provide an important window for cancer interception. This review examines how etiological exposures, persistent inflammation, and lesion-specific epithelial–stromal–immune [...] Read more.

Oral squamous cell carcinoma (OSCC) frequently develops within chronically injured oral mucosa and may be preceded by clinically recognizable oral potentially malignant disorders (OPMDs), which provide an important window for cancer interception. This review examines how etiological exposures, persistent inflammation, and lesion-specific epithelial–stromal–immune interactions cooperate during the transition from mucosal injury to dysplasia, carcinoma in situ, and invasive OSCC. Major carcinogenic exposures, including tobacco, alcohol, and areca nut, are considered together with context-dependent contributors such as microbial dysbiosis, viral infection, and immune-mediated epithelial injury. At the molecular level, inflammation-driven oral carcinogenesis involves cytokine and chemokine amplification, oxidative and nitrosative stress, NF-κB and STAT3 activation, the COX-2/PGE₂ axis, genomic instability, field cancerization, epithelial–stromal crosstalk, angiogenesis, immune dysregulation, and epigenetic and non-coding RNA-mediated reprogramming. Emerging tools such as molecular risk assessment, liquid biopsy, optical imaging, spatially resolved profiling, and artificial intelligence-assisted models may improve identification of high-risk lesions, although most biomarkers require further prospective validation. Prevention should therefore integrate exposure control, biopsy-based diagnosis, local treatment when indicated, long-term surveillance, and trial-based precision strategies according to lesion risk, intervention window, and safety profile. This review supports a shift from lesion-centered management toward risk-adapted precision prevention in inflammation-driven oral carcinogenesis. Full article

(This article belongs to the Section Molecular Pathology, Diagnostics, and Therapeutics)

► Show Figures

Figure 1

17 pages, 16014 KB

Open AccessArticle

Microbial Contamination, Degradation Characteristics of Dominant Bacteria on the Hull of the Nanhai No. 1 Shipwreck

by Yu Wang, Yeqing Han, Cen Wang, Zeao Wang, Zhiqian Guan, Naisheng Li and Jiao Pan

Int. J. Mol. Sci. 2026, 27(12), 5631; https://doi.org/10.3390/ijms27125631 - 22 Jun 2026

Viewed by 357

Abstract

To clarify the microbial contamination and wood degradation risk of the Nanhai No. 1 shipwreck hull and verify on-site antibacterial agent effectiveness, microbial samples were collected and analyzed via SEM, metagenomic sequencing, bacterial isolation, enzyme activity detection, and antibacterial experiments. The results showed [...] Read more.

To clarify the microbial contamination and wood degradation risk of the Nanhai No. 1 shipwreck hull and verify on-site antibacterial agent effectiveness, microbial samples were collected and analyzed via SEM, metagenomic sequencing, bacterial isolation, enzyme activity detection, and antibacterial experiments. The results showed that Actinomycetota was the dominant phylum, and Brachybacterium, Microbacterium, and Brevibacterium were the dominant genera. Seven bacterial strains were isolated and purified, among which Brevibacterium sp. (NH.SH-B6) had the strongest wood degradation ability, possessing cellulase, LiP, MnP, and Lac activities. When cultured with hull wood as the sole carbon source, LiP was the dominant degrading enzyme of NH.SH-B6, and its maximum enzyme activity was achieved under the optimal conditions of pH = 7, 10% NaCl, 1000 mg/L FeSO₄, and no PEG400 added. 50 mg/mL cinnamaldehyde and 0.5% isothiazolinone K100 had good inhibitory effects on the isolated bacteria, and bacterial proliferation was due to incomplete antibacterial agent spraying. This study clarifies the microbial degradation risk of the Nanhai No. 1 shipwreck hull and provides a scientific basis for optimizing the on-site protection strategy of the shipwreck. Full article

(This article belongs to the Section Molecular Microbiology)

► Show Figures

Figure 1

12 pages, 1312 KB

Open AccessArticle

Differential Modulation of GLP-1R by Dietary Ginsenosides Points to a Putative Extracellular Allosteric Site

by Ayelet Caspi, Netaly Khazanov, Aharon Helman, Hodaya Lankry, Berta Levavi-Sivan, Hanoch Senderowitz and Zohar Kerem

Int. J. Mol. Sci. 2026, 27(12), 5630; https://doi.org/10.3390/ijms27125630 - 22 Jun 2026

Viewed by 490

Abstract

The glucagon-like peptide-1 receptor (GLP-1R) is a class B G protein-coupled receptor (GPCR) central to metabolic regulation, and its potential modulation by dietary phytochemicals is increasingly recognized as physiologically relevant. Understanding how such compounds interact with GLP-1R is important for clarifying mechanisms that [...] Read more.

The glucagon-like peptide-1 receptor (GLP-1R) is a class B G protein-coupled receptor (GPCR) central to metabolic regulation, and its potential modulation by dietary phytochemicals is increasingly recognized as physiologically relevant. Understanding how such compounds interact with GLP-1R is important for clarifying mechanisms that may contribute to gut-to-brain signaling. In this study, we examined three structurally related dietary ginsenosides, Rg1, Rg2, and Rg3, as potential modulators of GLP-1R using luciferase reporter assays and computational analyses. Despite sharing similar molecular weights, a common dammarane scaffold, and comparable sugar moieties, the three ginsenosides displayed distinct effects on GLP-1R activity: Rg2 and Rg3 potently reduced receptor activation in a dose-dependent manner when co-administered with Exendin-4, whereas Rg1 had minimal effect. Computational screening of the GLP-1R structure for binding sites identified a putative extracellular pocket on the protein that can accommodate these compounds, while molecular docking and binding free energy calculations provided predicted affinities qualitatively reflecting the phytochemicals’ experimental activities. These findings point to a plausible extracellular mechanism through which dietary ginsenosides may influence GLP-1R responsiveness at the intestinal interface. Our results point to the possibility that non-absorbed phytochemicals can differentially modulate gut-expressed receptors, suggesting a novel pathway for dietary signaling relevant to ethnopharmacology and metabolic health. Full article

(This article belongs to the Special Issue Biosynthesis of Phytochemicals and Their Applications in Novel Drug Lead Development)

► Show Figures

Figure 1

21 pages, 30091 KB

Open AccessArticle

Comparative Analysis of Serum and Tissue miRNA Expression Profiles and Regulatory Pathways in Early-Stage Ovarian Cancer Using Public Databases

by Shuya Cai, Hui Tan, Xiaoyu Niu, Nirupal Eskar and Zaoling Liu

Int. J. Mol. Sci. 2026, 27(12), 5629; https://doi.org/10.3390/ijms27125629 - 22 Jun 2026

Viewed by 592

Abstract

To characterize the distinct expression profiles of microRNAs (miRNAs) in serum and tissue and to delineate the heterogeneity of their regulatory mechanisms in early-stage ovarian cancer (EOC), thereby identifying candidate biomarkers for non-invasive early diagnosis. Differentially expressed miRNAs were identified by integrating publicly [...] Read more.

To characterize the distinct expression profiles of microRNAs (miRNAs) in serum and tissue and to delineate the heterogeneity of their regulatory mechanisms in early-stage ovarian cancer (EOC), thereby identifying candidate biomarkers for non-invasive early diagnosis. Differentially expressed miRNAs were identified by integrating publicly available datasets of EOC tissues and serum samples from the Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA). Core miRNAs were subsequently screened through integrated differential expression analysis, weighted gene co-expression network analysis (WGCNA), and feature importance ranking derived from optimized machine learning models. Protein–protein interaction (PPI) networks and functional enrichment analyses (GO and KEGG) were performed on predicted target genes to systematically compare the functional discrepancies between serum- and tissue-derived miRNAs. No overlapping core miRNAs were observed between the two compartments. Serum miRNAs exhibited an overall up-regulated trend, whereas tissue miRNAs were predominantly down-regulated. Although the regulatory pathways demonstrated significant heterogeneity, they ultimately converged on the cell cycle and the PI3K-Akt signaling pathway, indicating high functional homology. Furthermore, serum miRNAs are not merely passive leakage products from tissues; current evidence suggests they may be selectively packaged into exosomes to participate in tumor regulation. Despite divergent expression profiles, serum and tissue miRNAs share homologous regulatory functions in EOC. These findings suggest that serum miRNAs accurately reflect the core molecular status of tumor tissues, providing a robust molecular foundation for liquid biopsy-based early detection strategies. Full article

(This article belongs to the Section Molecular Informatics)

► Show Figures

Figure 1

16 pages, 3340 KB

Open AccessArticle

Comparison of PBS-Caffeine and Caffeine Buffers for Inhibiting Exocytosis During Horseshoe Crab Blood Collection and Improving the Yield of Limulus Amebocyte Lysate (LAL) for Endotoxin Detection

by Jessica Zhang, Sophia Zhang and Mengmeng Zhang

Int. J. Mol. Sci. 2026, 27(12), 5628; https://doi.org/10.3390/ijms27125628 - 22 Jun 2026

Viewed by 329

Abstract

Limulus amebocyte lysate (LAL) detects bacterial endotoxin through a serine-protease coagulation cascade in which Factor C responds to lipopolysaccharide and Factor G to (1,3)-β-D-glucan. Sustainable LAL production depends on collection buffers that prevent amebocyte degranulation while preserving these clotting factors. We previously showed [...] Read more.

Limulus amebocyte lysate (LAL) detects bacterial endotoxin through a serine-protease coagulation cascade in which Factor C responds to lipopolysaccharide and Factor G to (1,3)-β-D-glucan. Sustainable LAL production depends on collection buffers that prevent amebocyte degranulation while preserving these clotting factors. We previously showed that caffeine buffer inhibits degranulation, but caffeine-collected pellets aggregated upon resuspension in 5 mM CaCl₂, unlike phosphate-buffered saline (PBS). We therefore developed a PBS-caffeine collection solution and compared it with caffeine buffer. Over one bleeding season, 121 crabs were bled; blood was collected in caffeine, PBS-caffeine, or PBS-caffeine supplemented with EDTA, EGTA, or both, and LAL activity was measured by chromogenic and turbidimetric assays. Both buffers prevented degranulation and gave comparable LAL activity, but PBS-caffeine reduced aggregation and clotting. Treating PBS-caffeine LAL with 10% PEG-8000 selectively abolished endotoxin-sensitive Factor C activity while preserving (1,3)-β-D-glucan–sensitive Factor G activity, and the resulting Factor G lysate, formulated in 20 mM acetate (pH 5.6), remained stable for 27 months. These results define an improved collection buffer and identify conditions that selectively stabilize Factor G zymogen in liquid form. Full article

(This article belongs to the Section Biochemistry)

► Show Figures

Figure 1

21 pages, 5296 KB

Open AccessArticle

IMMUND: A Diagnostic and Therapeutic Pipeline to Uncover the Convergence in Functional Perturbation at Early Stages of Neurodegenerative Diseases and Multiple Sclerosis Based on Protein Markers

by Ashmita Dey, Dwipanjan Sanyal, Krishnananda Chattopadhyay, Ujjwal Maulik, Vladimir N. Uversky and Sagnik Sen

Int. J. Mol. Sci. 2026, 27(12), 5627; https://doi.org/10.3390/ijms27125627 - 22 Jun 2026

Viewed by 911

Abstract

Neuroinflammation is a key hallmark of both neurodegenerative and neurospecific autoimmune diseases, including multiple sclerosis (MS), where immune dysregulation contributes to cellular stress, autophagy, and disease progression in Alzheimer’s disease (AD), Parkinson’s disease (PD), and MS. Emerging evidence suggests a shared mechanism behind [...] Read more.

Neuroinflammation is a key hallmark of both neurodegenerative and neurospecific autoimmune diseases, including multiple sclerosis (MS), where immune dysregulation contributes to cellular stress, autophagy, and disease progression in Alzheimer’s disease (AD), Parkinson’s disease (PD), and MS. Emerging evidence suggests a shared mechanism behind MS, AD, and PD, driven by chronic interaction between the peripheral immune system and the central nervous system (CNS). While MS was traditionally viewed as a primary autoimmune condition, recent research indicated that all three disorders involve a breakdown of the blood–brain barrier (BBB). This structural failure enables peripheral immune cells and cytokines to enter the brain, causing sustained neuroinflammation and accelerating disease progression. Here, we propose an end-to-end framework for identification of the diagnostic and therapeutic cell-specific protein markers commonly regulated in mild–moderate AD (MMAD), early-stage PD (ESPD), and MS within peripheral blood mononuclear cells (PBMCs). PBMC markers were first identified based on shared differential protein expression, followed by filtering for BBB permeability. Subsequently, sorted cell markers were mapped to disease-specific neural cell types. Our analysis suggests that PBMC-derived cells, including astrocyte- and monocyte-like populations, share overlapping transcriptional signatures and functional similarity with macrophages and neuroglial cells, indicating potential transcriptional similarity or functional convergence. Furthermore, intra- and inter-cellular pathway analysis suggested both shared and disease-specific signaling mechanisms, with kinase–integrin interactions emerging as key regulatory factors. Selected potential seed markers, primarily kinases and immunoglobulins, were further analyzed through evolutionary sequence–structure space to identify druggable structural features. Next, protein moonlighting possibilities were tested to enhance the temporal functional trajectory of the markers for precise therapeutic impact. Hence, the framework provides a robust strategy to identify immune-based disease-specificcandidate diagnostic andpotential therapeutic targets. Full article

(This article belongs to the Topic Applications of Biomedical Technology and Molecular Biological Approach in Brain Diseases, 2nd Edition)

► Show Figures

Figure 1

61 pages, 12191 KB

Open AccessReview

Curcumin and Its Derivatives as Anticancer Agents in Head and Neck Cancer: Molecular Mechanisms and Preclinical Evidence

by Luana Pinto, João P. N. Silva, Luís Monteiro and Patrícia M. A. Silva

Int. J. Mol. Sci. 2026, 27(12), 5626; https://doi.org/10.3390/ijms27125626 - 22 Jun 2026

Cited by 2 | Viewed by 700

Abstract

Head and neck cancer (HNC), particularly oral squamous cell carcinoma (OSCC), remains a major clinical challenge due to its aggressive behavior, high recurrence rates, and frequent resistance to conventional therapies. Natural compounds, especially curcumin and its derivatives, have gained increasing attention as potential [...] Read more.

Head and neck cancer (HNC), particularly oral squamous cell carcinoma (OSCC), remains a major clinical challenge due to its aggressive behavior, high recurrence rates, and frequent resistance to conventional therapies. Natural compounds, especially curcumin and its derivatives, have gained increasing attention as potential anticancer agents due to their ability to target multiple molecular pathways involved in tumor progression. This review critically evaluates the current preclinical and translational evidence supporting curcumin and its derivatives as monotherapeutic agents in HNC, with particular emphasis on oral cancer. We integrate the available evidence to assess the biological rationale, therapeutic potential, and current limitations of curcumin-based approaches. The molecular mechanisms underlying their antitumor activity are discussed, including modulation of EGFR/ERK and PI3K/Akt signaling pathways, inhibition of NF-κB and STAT3 activation, induction of apoptosis, regulation of oxidative stress, and suppression of epithelial–mesenchymal transition and tumor invasiveness. In addition, we address the impact of curcumin on the tumor microenvironment and its role in overcoming intrinsic cellular resistance mechanisms. The review also highlights advances in drug delivery strategies, such as nanoformulations, that are designed to improve curcumin’s bioavailability and therapeutic efficacy. By critically integrating current evidence, this review highlights both the promise and the challenges associated with curcumin-based monotherapy in HNC, emphasizing the need for more robust and clinically relevant studies to support future translation. Full article

(This article belongs to the Special Issue Bioactive Compounds in Cancers: Second Edition)

► Show Figures

Figure 1

19 pages, 1390 KB

Open AccessReview

Deubiquitinating Enzymes as Therapeutic Candidates in Hepatocellular Carcinoma and Other Liver Disease

by Young-Hoon Jeong, Hwa-Hyeong Lee, Young-Jun Kim, Hye-Rim Lee and Key-Hwan Lim

Int. J. Mol. Sci. 2026, 27(12), 5625; https://doi.org/10.3390/ijms27125625 - 22 Jun 2026

Viewed by 572

Abstract

Hepatocellular carcinoma is challenging to detect at an early stage, and its severity increases over time. Recently, the incidence of hepatocellular carcinoma has increased, partly due to lifestyle-related factors such as excessive alcohol intake, sedentary behavior, and diets high in fat, which contribute [...] Read more.

Hepatocellular carcinoma is challenging to detect at an early stage, and its severity increases over time. Recently, the incidence of hepatocellular carcinoma has increased, partly due to lifestyle-related factors such as excessive alcohol intake, sedentary behavior, and diets high in fat, which contribute to the growing prevalence of fatty liver and hepatitis. Various therapeutic strategies are being explored for hepatocellular carcinoma, among which therapies targeting deubiquitinating enzymes (DUBs) have attracted growing attention. Ubiquitination acts as a crucial modulator in the regulation of intracellular signaling across many diseases. E3 ligase recognizes the target protein and transfers ubiquitin, received from the E2 enzyme, to the lysine residues of the substrate, thereby conferring specificity to the ubiquitination process. Once a ubiquitin chain is attached to a target protein by an E3 ligase, the protein is directed to the ubiquitin–proteasome system (UPS) for degradation. In this process, the 26S proteasome complex recognizes the ubiquitin chain and degrades the target protein, thereby serving as a major mechanism for maintaining protein homeostasis. Through this pathway, cells regulate signal transduction, eliminate abnormal proteins, and perform various essential functions. On the other hand, deubiquitinating enzymes (DUBs) recognize the ubiquitin chains on target proteins and remove them by hydrolyzing the isopeptide bonds of ubiquitin, thereby enabling the target proteins to evade degradation by the proteasome system. Furthermore, deubiquitinating enzymes independently remove ubiquitin from proteins and can serve as central regulators in signaling pathways related to hepatocellular carcinoma. Full article

(This article belongs to the Special Issue Liver Diseases: From Pathophysiology to Novel Therapeutic Approaches)

► Show Figures

Figure 1

20 pages, 4507 KB

Open AccessArticle

Liraglutide, a GLP-1 Receptor Agonist, Mitigates LPS-Induced Osteoclastogenesis and Bone Loss by Downregulating Macrophage TNF-α Expression

by Kou Murakami, Hideki Kitaura, Fumitoshi Ohori, Aseel Marahleh, Angyi Lin, Ziqiu Fan, Kohei Narita, Tomoko Ishiyama, Jin Hu, Huidan Zheng and Hiroyasu Kanetaka

Int. J. Mol. Sci. 2026, 27(12), 5624; https://doi.org/10.3390/ijms27125624 - 22 Jun 2026

Cited by 1 | Viewed by 653

Abstract

Liraglutide, a glucagon-like peptide-1 (GLP-1) receptor agonist, restores hyperglycemic conditions in patients with type 2 diabetes and has recently shown promising anti-inflammatory properties. In this study, we explored its potential to suppress osteoclast formation and bone loss triggered by lipopolysaccharide (LPS), an inflammatory [...] Read more.

Liraglutide, a glucagon-like peptide-1 (GLP-1) receptor agonist, restores hyperglycemic conditions in patients with type 2 diabetes and has recently shown promising anti-inflammatory properties. In this study, we explored its potential to suppress osteoclast formation and bone loss triggered by lipopolysaccharide (LPS), an inflammatory agent. In animal models, the co-administration of liraglutide with LPS on the calvaria regions in mice markedly reduced osteoclast numbers and bone resorption areas relative to treatment with LPS alone. Furthermore, the expression levels of receptor activators of the NF-κB ligand (RANKL) and tumor necrosis factor (TNF)-α mRNA were notably lower in the group receiving liraglutide and LPS compared to treatment with LPS alone. Moreover, in vitro tests revealed that liraglutide has no direct inhibitory effect on RANKL-induced osteoclastogenesis and TNF-α-induced osteoclastogenesis. In addition, liraglutide had no direct inhibitory effect on LPS-stimulated RANKL expression in osteoblasts. Moreover, liraglutide effectively suppressed TNF-α mRNA expression in macrophages stimulated by LPS. These findings suggest that liraglutide prevents inflammatory bone destruction not by targeting osteoclast formation directly but by inhibiting the production of TNF-α within macrophages. Full article

(This article belongs to the Special Issue Metabolic Regulators of Bone Health)

► Show Figures

Figure 1

19 pages, 2746 KB

Open AccessReview

A Systematic Review on the Association Between Water Fluoride Levels and Dental Fluorosis: Exploring the ‘Halo Effect’ and Confounding Environmental Factors

by Mnqweno Funcuza, Bheki T. Magunga, Phoka C. Rathebe and Thokozani P. Mbonane

Int. J. Mol. Sci. 2026, 27(12), 5623; https://doi.org/10.3390/ijms27125623 - 22 Jun 2026

Viewed by 964

Abstract

Dental fluorosis (DF) remains a global public health challenge traditionally attributed to elevated water fluoride F⁻. However, the Halo Effect and environmental factors now complicate this dose–response relationship. Following PRISMA 2020 guidelines, this systematic review identified 20 observational studies (n [...] Read more.

Dental fluorosis (DF) remains a global public health challenge traditionally attributed to elevated water fluoride F⁻. However, the Halo Effect and environmental factors now complicate this dose–response relationship. Following PRISMA 2020 guidelines, this systematic review identified 20 observational studies (n = 21,780) via PubMed, Scopus, and Web of Science. Inclusion logic utilized the PICOS framework, specifically selecting human studies that reported quantitative water F⁻ levels alongside environmental or dietary confounders. Quality was assessed via the Newcastle–Ottawa Scale. Synthesis revealed that in optimal fluoridated areas (0.7 mg/L), mild DF prevalence reached 15–20% in cohorts with high “Halo Effect” exposure (infant formula, processed beverages) a twofold increase over historical benchmarks. High altitude (>2000 m) and arid climates further exacerbated toxicity by altering renal clearance. These factors sustain systemic fluoride levels that inhibit protease activity (MMP-20/KLK4) and induce endoplasmic reticulum stress during enamel maturation, causing hypomineralization. Current water-centric monitoring is insufficient for modern risk assessment. A transition toward Total Daily Intake (TDI) models and context-specific standards accounting for altitude and dietary diffusion is essential to balance caries prevention with systemic safety. Full article

(This article belongs to the Special Issue Oral and Dental Infectious Diseases: Molecular Pathology, Diagnosis and Therapy (2nd Edition))

► Show Figures

Figure 1

28 pages, 23802 KB

Open AccessArticle

Synergistic Induction of Caspase-8-Mediated Leukaemic Cell Death by Fisetin and Pinocembrin

by Narawan Kaewthawee, Ankita Sharma, James Michael Brimson and Sirikalaya Brimson

Int. J. Mol. Sci. 2026, 27(12), 5622; https://doi.org/10.3390/ijms27125622 - 22 Jun 2026

Viewed by 517

Abstract

Fisetin is a bioactive flavanol with reported anticancer activity, although its mechanisms in leukaemia and potential for combination therapy remain incompletely understood. This study investigated the cytotoxic and mechanistic effects of fisetin, alone and combined with pinocembrin, in human leukaemia cells. Cell viability, [...] Read more.

Fisetin is a bioactive flavanol with reported anticancer activity, although its mechanisms in leukaemia and potential for combination therapy remain incompletely understood. This study investigated the cytotoxic and mechanistic effects of fisetin, alone and combined with pinocembrin, in human leukaemia cells. Cell viability, apoptosis, and cell cycle progression were assessed by flow cytometry; protein expression in Jurkat cells was assessed by Western blotting; and molecular docking was used to evaluate interactions with the Fas receptor. Drug interactions were quantified using ZIP synergy analysis, and cytotoxicity and clonogenic survival were evaluated using soft-agar colony formation assays in K562 cells. Fisetin significantly reduced cell viability and induced apoptosis, accompanied by caspase-8 cleavage, p62 accumulation, and CDK4 downregulation, consistent with activation of extrinsic apoptosis, impaired autophagic flux, and cell cycle inhibition in Jurkat cells. Docking analysis supported a potential interaction with the Fas receptor, which was confirmed using the Fas receptor antagonist Met-12. Co-treatment with pinocembrin enhanced fisetin-mediated cytotoxicity and produced synergistic effects, particularly in Jurkat cells (ZIP score > 10), while synergistic interactions at specific sub-IC₅₀ concentrations were also observed in K562 cells. Combination treatment further enhanced caspase-8 activation, reduced CDK4 expression in Jurkat cells, and significantly suppressed clonogenic survival in K562 cells compared with single-agent treatments. These findings suggest that fisetin promotes caspase-8-dependent apoptosis, potentially involving Fas-associated signalling, and highlight fisetin–pinocembrin combination therapy as a promising strategy for leukaemia treatment. Full article

(This article belongs to the Special Issue Biological Applications, Theoretical Studies, Nanomedicine and Molecular Docking Studies in Drug Discovery)

► Show Figures

Figure 1

33 pages, 4421 KB

Open AccessArticle

Effects of Nonionizing Millimeter-Wave on Spheroid-like Irradiated Non-Small-Cell Lung Cancer (NSCLC) Cells

by Helena Tuchinsky, Boris Litvak, Vladimir Freydin, Firas Simaan, Rawad Said, Dhaval Patel, Yosef Pinhasi, Asher Yahalom and Stella Liberman-Aronov

Int. J. Mol. Sci. 2026, 27(12), 5621; https://doi.org/10.3390/ijms27125621 - 22 Jun 2026

Viewed by 665

Abstract

Non-thermal millimeter-wave (MMW) irradiation represents a promising non-invasive strategy for cancer therapy, yet its effects in physiologically relevant 3D systems remain poorly defined. Here, we evaluated the biological impact of MMW exposure in 3D non-small-cell lung cancer (NSCLC) spheroids (NCI-H1299, A549) and normal [...] Read more.

Non-thermal millimeter-wave (MMW) irradiation represents a promising non-invasive strategy for cancer therapy, yet its effects in physiologically relevant 3D systems remain poorly defined. Here, we evaluated the biological impact of MMW exposure in 3D non-small-cell lung cancer (NSCLC) spheroids (NCI-H1299, A549) and normal WI-38 fibroblasts under active cooling to suppress bulk heating. We demonstrate that cellular responses are governed primarily by power density (PD), irradiation geometry, and genotype-dependent susceptibility. High-PD pyramidal horn (PH) irradiation (~4.9 mW/cm²) induced rapid apoptosis, metabolic collapse, and near-complete loss of clonogenic survival, whereas lower-PD waveguide (WG) irradiation (~0.6 mW/cm²) produced depth-limited, cumulative cytotoxicity. Surviving cancer cells exhibited robust senescence-associated growth arrest, particularly in p53-deficient NCI-H1299 cells, indicating a dual apoptotic–senescent anti-proliferative response. In contrast, WI-38 fibroblasts showed minimal apoptosis and only transient stress-associated senescence, confirming selective tumor vulnerability. Mechanistic modeling suggests that MMW energy couples to glycan-rich membrane domains, generating localized electromagnetic hotspots that trigger calcium influx, mitochondrial dysfunction, and depth-dependent apoptosis. These findings establish a mechanistic basis for selective, non-thermal MMW-induced cytotoxicity in 3D NSCLC models and support further preclinical development of MMW-based therapeutic strategies. Full article

(This article belongs to the Special Issue Radiofrequency on Human Health: A Molecular Perspective)

► Show Figures

Graphical abstract

28 pages, 1512 KB

Open AccessReview

Isothiocyanates as Multi-Target Natural Compounds in Leukemia: Mechanisms, Selectivity, and Therapeutic Potential

by Alberto Yoldi Vergara, Kristina Simonicova, Anna Bertova, Zdena Sulova, Albert Breier and Denisa Imrichova

Int. J. Mol. Sci. 2026, 27(12), 5620; https://doi.org/10.3390/ijms27125620 - 22 Jun 2026

Viewed by 672

Abstract

Natural compounds are increasingly explored as complementary strategies to enhance the effectiveness of chemotherapy and reduce toxicity. Among these are isothiocyanates (ITCs), bioactive metabolites derived from glucosinolates in cruciferous vegetables, which have gained substantial attention for their chemopreventive and antileukemic potential. ITCs exert [...] Read more.

Natural compounds are increasingly explored as complementary strategies to enhance the effectiveness of chemotherapy and reduce toxicity. Among these are isothiocyanates (ITCs), bioactive metabolites derived from glucosinolates in cruciferous vegetables, which have gained substantial attention for their chemopreventive and antileukemic potential. ITCs exert diverse biological effects driven by the high reactivity of the –NCS group, enabling covalent modification of key cellular proteins and modulation of signaling pathways. Well-studied representatives, including sulforaphane (SFN), allyl isothiocyanate (AITC), 6-(methylsulfinyl)hexyl isothiocyanate (6-MITC), benzyl isothiocyanate (BITC), and phenethyl isothiocyanate (PEITC), exhibit diverse antileukemic activities, including cytotoxic, pro-apoptotic, differentiation-inducing, and cell-cycle-modulating effects. Although individual compounds differ in their relative potency and predominant biological responses, their activities are generally mediated through multiple interconnected mechanisms including oxidative stress modulation, mitochondrial dysfunction, regulation of apoptosis-related proteins, and interference with key signaling pathways. In addition to apoptosis, several ITCs have also been reported to induce autophagy, ferroptosis, or cellular differentiation in leukemic cells. Taken together, the existing evidence highlights ITCs as promising candidates for leukemia chemoprevention or therapy, acting through multi-targeted mechanisms that may complement conventional treatment strategies. Further studies are needed to clarify their selectivity, mechanistic diversity, and translational potential. Full article

(This article belongs to the Special Issue Precision Nutrition and Cancer: Molecular Insights into Cancer Prevention and Development)

► Show Figures

Graphical abstract

25 pages, 3598 KB

Open AccessArticle

Aortic Single-Cell Transcriptome Analysis Reveals ApoE-Isoform-Specific Influences on Vascular Disease

by David Y. Hui, Jeyashree Alagarsamy, April Haller, Mario Medvedovic and Anja Jaeschke

Int. J. Mol. Sci. 2026, 27(12), 5619; https://doi.org/10.3390/ijms27125619 - 22 Jun 2026

Viewed by 571

Abstract

The human APOE gene is polymorphic with three major alleles that encode apolipoprotein (apo) E2, apoE3, and apoE4. Both apoE2 and apoE4 are associated with increased atherosclerosis risk. This study utilized human APOE2, APOE3, and APOE4 gene replacement mice and single-cell [...] Read more.

The human APOE gene is polymorphic with three major alleles that encode apolipoprotein (apo) E2, apoE3, and apoE4. Both apoE2 and apoE4 are associated with increased atherosclerosis risk. This study utilized human APOE2, APOE3, and APOE4 gene replacement mice and single-cell RNA sequencing approach to delineate the mechanisms underlying this association. The human APOE2, APOE3, and APOE4 mice were fed a Western-type high fat–cholesterol diet for 16 weeks. Hyperlipidemia and significant atherosclerosis were observed in APOE2 mice but not in APOE3 or APOE4 mice. However, increased vascular inflammation was observed in both APOE2 and APOE4 mice. Single-cell RNA sequencing followed by cluster analysis identified 25 major cell types in the aorta that include various immune cell types, endothelial cells, and various vascular mural cell subsets. Results showed that cells from the APOE2 mice were enriched with genes associated with intracellular lipid accumulation and inflammation, whereas cells from the APOE4 mice displayed elevated oxidative- and/or endoplasmic reticulum-stress and inflammatory response. Thus, apoE2 accelerates atherosclerosis by inducing diet-induced hyperlipidemia and inflammation, while apoE4 does not induce hyperlipidemia but enhances inflammation that may prime the vasculature for atherosclerosis development. The distinct mechanisms by which apoE2 and apoE4 promote atherosclerosis underscore the importance of including apoE genotype information in the design of therapeutics for atherosclerosis intervention. Full article

(This article belongs to the Special Issue Atherosclerosis: From Molecular Mechanisms, Pathophysiology to Novel Therapeutic Approaches (3rd Edition))

► Show Figures

Figure 1

24 pages, 1016 KB

Open AccessReview

Therapeutic Effects of Glucagon-like Peptide-1 Receptor Agonists in Non-Alcoholic Fatty Liver Disease: A Systematic Review

by Dina Mahoon, Fares Kellany, Imad Khan, Somieya Khan and Alexandra E. Butler

Int. J. Mol. Sci. 2026, 27(12), 5618; https://doi.org/10.3390/ijms27125618 - 22 Jun 2026

Cited by 1 | Viewed by 1626

Abstract

Non-alcoholic fatty liver disease (NAFLD), now increasingly termed metabolic dysfunction-associated steatotic liver disease (MASLD), is a growing cause of chronic liver disease with limited treatment options. Glucagon-like peptide-1 (GLP-1) receptor agonists, approved for type 2 diabetes and obesity, possess metabolic effects that may [...] Read more.

Non-alcoholic fatty liver disease (NAFLD), now increasingly termed metabolic dysfunction-associated steatotic liver disease (MASLD), is a growing cause of chronic liver disease with limited treatment options. Glucagon-like peptide-1 (GLP-1) receptor agonists, approved for type 2 diabetes and obesity, possess metabolic effects that may render them suitable for treating NAFLD and metabolic dysfunction-associated steatohepatitis (MASH). To evaluate the therapeutic effects of GLP-1 receptor agonists in adults with NAFLD, non-alcoholic steatohepatitis (NASH), MASLD, or MASH. PubMed, Scopus, Embase, and the Cochrane Library were systematically searched using keywords related to NAFLD and GLP-1 receptor agonists. Given heterogeneity in populations, designs, and outcomes, findings were synthesized narratively. The review is registered with PROSPERO (CRD420261337353). Twelve studies met the inclusion criteria. The most consistent outcome was a reduction in hepatic fat, seen with semaglutide, liraglutide, dulaglutide, and beinaglutide. Improvements in liver enzymes, particularly alanine aminotransferase, were less consistent and best regarded as supportive rather than definitive evidence of histological improvement. Histological benefits were strongest for steatohepatitis resolution in non-cirrhotic MASH. Fibrosis findings were mixed, with the greatest benefit in F2–F3 MASH and limited improvement in established cirrhosis. GLP-1 receptor agonists were generally well tolerated, with gastrointestinal symptoms the most common adverse effects. GLP-1 receptor agonists show promising liver-related benefits in NAFLD and MASH, particularly in obesity, type 2 diabetes, or earlier-stage disease. Their effects on advanced fibrosis and long-term outcomes remain uncertain, warranting larger, longer-term studies. Full article

(This article belongs to the Special Issue Molecular and Translational Research in Cardiovascular Endocrinology, Cardiovascular–Kidney–Metabolic Diseases and Metabolic Dysfunction-Associated Steatotic Liver Disease (MASLD): 3rd Edition)

► Show Figures

Figure 1

17 pages, 8032 KB

Open AccessArticle

Associative Analysis of lncRNA/circRNA-miRNA-mRNA Expression Profiles in Iron-Overloaded HT-1080 Fibrosarcoma Cells

by Yifan Teng, Qian Zhang, Haoxuan Ding and Jie Feng

Int. J. Mol. Sci. 2026, 27(12), 5617; https://doi.org/10.3390/ijms27125617 - 22 Jun 2026

Viewed by 410

Abstract

Iron overload disrupts cellular homeostasis and drives ferroptosis through dysregulated iron metabolism. Non-coding RNAs (ncRNAs) are considered as key regulators of various biological functions and targets for a new generation of RNA therapeutics and biomarkers. However, few studies have investigated the regulatory roles [...] Read more.

Iron overload disrupts cellular homeostasis and drives ferroptosis through dysregulated iron metabolism. Non-coding RNAs (ncRNAs) are considered as key regulators of various biological functions and targets for a new generation of RNA therapeutics and biomarkers. However, few studies have investigated the regulatory roles of ncRNAs, particularly competitive endogenous RNAs (ceRNAs) in iron overload. This study performed whole-transcriptome sequencing to characterize the ceRNA network in ferric ammonium citrate (FAC)-induced iron-overloaded HT-1080 fibrosarcoma cells. A total of 208 differentially expressed mRNAs, 83 lncRNAs, and 170 circRNAs (q < 0.05) were identified, with hierarchical clustering revealing distinct expression patterns between control and iron-treated groups. KEGG enrichment implicated vitamin B6 metabolism (q < 0.001) and lysine degradation (q < 0.001) as key disrupted pathways. ceRNA network was conducted and further demonstrated lncRNA/circRNA-mediated regulation of ferroptosis genes via shared miRNA response elements. Notably, LINC-PINT-232 was implicated in the regulation of both ferritin heavy chain (FTH) and sequestosome 1 (SQSTM1), two ferroptosis-associated mRNAs. FTH upregulation mitigates iron toxicity through ferroxidase activity, while SQSTM1 modulates lipid peroxidation in ferroptosis. These findings provide a preliminary transcriptomic landscape for hypothesis generation regarding ncRNA-mediated regulatory mechanisms in iron overload-induced ferroptosis and offer a computational foundation for future functional and therapeutic investigations. Full article

(This article belongs to the Special Issue The Role of Trace Elements in Nutrition and Health, 2nd Edition)

► Show Figures

Figure 1

31 pages, 1326 KB

Open AccessReview

Bidirectional Interactions Between Cervicovaginal Microbiota and Human Papillomavirus Drive Persistence and Disease Progression

by Daniel Osmar Suárez-Rico, Lourdes del Carmen Rizo de la Torre, Martin Zermeño-Ruiz, Luis Ricardo Balleza-Alejandri, Jesús Jonathan García-Galindo, Héctor Montoya-Fuentes and Alberto Beltrán-Ramírez

Int. J. Mol. Sci. 2026, 27(12), 5616; https://doi.org/10.3390/ijms27125616 - 22 Jun 2026

Cited by 1 | Viewed by 642

Abstract

Persistent high-risk human papillomavirus infection is a critical prerequisite for cervical intraepithelial neoplasia and cervical cancer, yet viral factors alone do not fully explain why most infections clear while a subset persists and progresses. Emerging longitudinal, multi-omics, and mechanistic evidence supports a plausible [...] Read more.

Persistent high-risk human papillomavirus infection is a critical prerequisite for cervical intraepithelial neoplasia and cervical cancer, yet viral factors alone do not fully explain why most infections clear while a subset persists and progresses. Emerging longitudinal, multi-omics, and mechanistic evidence supports a plausible model in which the cervicovaginal microbiota is not a passive bystander but a functional determinant of mucosal immunity, epithelial barrier integrity, and local metabolic tone. Lactobacillus-dominant community states, particularly those enriched in Lactobacillus crispatus, are generally associated with lower pH, regulated inflammatory signaling, stronger barrier function, and a higher likelihood of HPV clearance. In contrast, anaerobe-enriched dysbiosis is linked to elevated pro-inflammatory cytokines, altered antigen presentation, immune checkpoint signatures consistent with T-cell dysfunction, and metabolic shifts involving lactate depletion and accumulation of short-chain fatty acids and other metabolites that can influence epithelial and immune-cell programs. Importantly, the interaction is bidirectional: hrHPV can remodel the microenvironment by suppressing host defense peptides and perturbing mucosal barriers, thereby reducing Lactobacillus fitness and reinforcing dysbiosis in a feed-forward loop that favors persistence and oncogenic progression. This review integrates functional ecology, longitudinal clinical evidence, immunological and metabolic mechanisms, and translational implications, highlighting opportunities for microbiome-informed risk stratification and adjunctive interventions, as well as key gaps requiring standardized longitudinal multi-omics and rigorously designed clinical trials. Full article

(This article belongs to the Special Issue Interactions Between Microbes and Hosts: Physiology, Pathology and Treatment—Second Edition)

► Show Figures

Figure 1

18 pages, 15648 KB

Open AccessArticle

Early Flowering (ELF) Gene Integrates Vegetative Growth, Flowering Regulation, and Reproductive Development in Arabidopsis thaliana

by Rahmatullah Jan, Shahzad Iqbal, Sajad Ali, Mohammed A. Almalki, Mohammad Alfredan, Rashid Ismael Hag Ibrahim, Sajjad Asaf and Kyung-Min Kim

Int. J. Mol. Sci. 2026, 27(12), 5615; https://doi.org/10.3390/ijms27125615 - 22 Jun 2026

Viewed by 500

Abstract

Early flowering-related factors play pivotal roles in coordinating plant growth and reproductive development. In this study, we investigated the biological function of early flowering gene (ELF) in Arabidopsis thaliana using CRISPR/Cas9-mediated genome editing and construction of overexpression approaches. Two independent ELF [...] Read more.

Early flowering-related factors play pivotal roles in coordinating plant growth and reproductive development. In this study, we investigated the biological function of early flowering gene (ELF) in Arabidopsis thaliana using CRISPR/Cas9-mediated genome editing and construction of overexpression approaches. Two independent ELF overexpression (OE-ELF) and genome-edited (ge-elf) lines were generated and systemically analyzed. ELF overexpression significantly enhanced early seedling performance, increasing germination rate and seedling fresh weight by up to 8.7%, while genome-edited lines exhibited a marked reduction. Root growth was strongly promoted in OE-ELF plants, with root length increase of 85% and 75%, whereas ge-elf lines showed a reduction of up to 48%. At later developmental stages, OE-ELF plants displayed enhanced vegetative growth, including increased leaf length (32%), leaf area (91%), and accelerated flowering (21% earlier than wild type). In contrast, ge-elf delayed flowering by up to 25% and resulted in compact plant architecture. Reproductive development was severely compromised in ge-elf plants, which exhibited malformed inflorescences, reduced pollen germination, shortened silique (45%), and a drastic decrease in seed number per silique (70%). Conversely, OE-ELF plants showed increased silique number and seed per silique. Molecular analysis revealed that ELF positively regulates key flowering-related genes, including FLC, SOC1, AP1, and LFY, which correlated strongly with growth and reproductive traits. Our results demonstrate that ELF functions as a central regulator integrating vegetative growth, floral development, male fertility, and seed production in Arabidopsis thaliana. Full article

(This article belongs to the Section Molecular Plant Sciences)

► Show Figures

Figure 1

30 pages, 4590 KB

Open AccessReview

Building Disease Models for Endometriosis: iPSCs as Game-Changers

by Khalisa H. Kahar, Bushra E-Anjum, Fazlina Nordin, Angela Min Hwei Ng, Nor Haslinda Abd Aziz, Izyan Mohd Idris, Gee Jun Tye and Wan Safwani Wan Kamarul Zaman

Int. J. Mol. Sci. 2026, 27(12), 5614; https://doi.org/10.3390/ijms27125614 - 22 Jun 2026

Viewed by 713

Abstract

This review aims to evaluate the potential of endometriosis models, especially patient-derived iPSC models, to gain deeper insights into the disease, thereby advancing our understanding and treatment of endometriosis. This comprehensive narrative review utilized a structured search of the PubMed, Scopus, and Web [...] Read more.

This review aims to evaluate the potential of endometriosis models, especially patient-derived iPSC models, to gain deeper insights into the disease, thereby advancing our understanding and treatment of endometriosis. This comprehensive narrative review utilized a structured search of the PubMed, Scopus, and Web of Science databases, primarily covering literature published between January 2000 and May 2025. An expansive search strategy was employed to capture the full breadth of the field using keywords such as “endometriosis,” “induced pluripotent stem cells (iPSCs),” “patient-derived organoids,” “disease modeling,” and “epigenetics” without restrictive filtering, ensuring the integration of both foundational theories and emerging biotechnological advances. In total, over 170 peer-reviewed publications were analyzed, ranging from landmark genomic meta-analyses that have identified significant risk loci to state-of-the-art 3D-culture systems for modeling patient-specific endometrial disease. By synthesizing these diverse sources, the review bridges the gap between traditional anatomical classifications and modern molecular modeling to evaluate the potential of iPSC platforms for personalized medicine and therapeutic discovery. Endometriosis is a multifactorial gynecological condition that affects 176 million women worldwide and can significantly impair quality of life. It occurs when endometrium-like tissue grows outside the uterus, responsive to ovarian hormones, causing inflammation, pain, and discomfort, and leading to fibrotic tissue. World Health Organization estimates indicate that 6–10% of women suffer from this disorder, which can cause infertility and increase the risk of developing various types of cancer and autoimmune disorders. The use of patient-derived iPSC models serves to gain deeper insights into the disease by mimicking the endometrial tissue or lesions observed in affected individuals, thereby advancing our understanding and treatment of endometriosis. Full article

(This article belongs to the Special Issue Research Progress on Disease Models Based on Induced Pluripotent Stem Cells)

► Show Figures

Graphical abstract

11 pages, 4889 KB

Open AccessArticle

High-Level Secretory Expression of Recombinant Type XVII Human-like Collagen in Komagataella phaffii

by Yixuan Jia, Junhao Yue, Wanting Wu, Weirui Zhao, Sheng Hu, Lehe Mei, Peilian Wei and Changjiang Lyu

Int. J. Mol. Sci. 2026, 27(12), 5613; https://doi.org/10.3390/ijms27125613 - 22 Jun 2026

Viewed by 528

Abstract

Type XVII collagen (COL17) is crucial for skin integrity but difficult to produce. To achieve high-level secretory expression, a human COL17 segment was designed and cloned into the pPIC9K vector with six different α-mating factor signal peptides and integrated into Komagataella phaffii GS115. [...] Read more.

Type XVII collagen (COL17) is crucial for skin integrity but difficult to produce. To achieve high-level secretory expression, a human COL17 segment was designed and cloned into the pPIC9K vector with six different α-mating factor signal peptides and integrated into Komagataella phaffii GS115. Fed-batch fermentation in a 5 L bioreactor yielded 4.7 g/L of recombinant COL17. Functional assays showed that it promoted the proliferation of human skin fibroblast (HFF) and immortalized keratinocytes (HaCaT), upregulated COL1A1, COL3A1, and TIMP1 in HFF cells, and enhanced skin barrier-related genes (KRT1, KRT5, KRT10, KRT14, IVL, LOR, FLG) in HaCaT cells. In a UVB-induced photoaging model, COL17 reduced reactive oxygen species (ROS) and matrix metalloproteinase 3 (MMP3) activity. This recombinant collagen exhibits photoprotective, regenerative, and barrier-enhancing activities, offering potential for skincare and tissue engineering. Full article

(This article belongs to the Section Molecular Microbiology)

► Show Figures

Graphical abstract

30 pages, 1372 KB

Open AccessReview

The Versatile Applications of Antisense Oligonucleotides in Modern Medicine

by Xue-Hai Liang and Lingdi Zhang

Int. J. Mol. Sci. 2026, 27(12), 5612; https://doi.org/10.3390/ijms27125612 - 22 Jun 2026

Cited by 1 | Viewed by 1282

Abstract

Antisense oligonucleotides (ASOs) are a class of nucleic acid therapeutics that modulate gene expression through diverse mechanisms. Since their initial demonstration in inhibiting viral genes, advances in medicinal chemistry, pharmacology, and delivery have enabled robust and durable target engagement across multiple tissues. Chemical [...] Read more.

Antisense oligonucleotides (ASOs) are a class of nucleic acid therapeutics that modulate gene expression through diverse mechanisms. Since their initial demonstration in inhibiting viral genes, advances in medicinal chemistry, pharmacology, and delivery have enabled robust and durable target engagement across multiple tissues. Chemical modifications to the backbone, ribose, and nucleobases have improved nuclease resistance, binding affinity, and pharmacokinetics, while conjugation and delivery technologies have expanded tissue accessibility. Beyond classical RNase H–mediated RNA degradation, ASOs regulate gene expression via splicing modulation, microRNA inhibition, transcriptional activation, and translation modulation, supporting both gene silencing and upregulation strategies. Multiple ASO drugs are now approved, particularly for genetic diseases, with many more in clinical development. This review outlines the evolution of antisense technology, key chemical and delivery innovations, ASO pharmacokinetics and intracellular trafficking, the mechanisms underlying gene regulation, and current clinical applications and future opportunities. Full article

(This article belongs to the Special Issue Antisense Oligonucleotides: Versatile Tools with Broad Applications)

► Show Figures

Figure 1

16 pages, 8698 KB

Open AccessArticle

Depot-Specific White Adipose Tissue Remodeling Supports Non-Thermogenic Metabolic Homeostasis During Shallow Hibernation in Raccoon Dogs

by Ruojun Zong, Zhiqiang Han, Runzhou Liu, Manman Yang, Xin Liu, Xiuli Zhang, Jiahao Hu, Rui Du and Chao Xu

Int. J. Mol. Sci. 2026, 27(12), 5611; https://doi.org/10.3390/ijms27125611 - 22 Jun 2026

Viewed by 431

Abstract

White adipose tissue (WAT) is essential for maintaining energy homeostasis during hibernation by supplying lipolysis-derived fatty acids as a major fuel source. In raccoon dogs (Nyctereutes procyonoides), the activity of brown adipose tissue is diminished, providing a unique model to investigate [...] Read more.

White adipose tissue (WAT) is essential for maintaining energy homeostasis during hibernation by supplying lipolysis-derived fatty acids as a major fuel source. In raccoon dogs (Nyctereutes procyonoides), the activity of brown adipose tissue is diminished, providing a unique model to investigate how WAT supports metabolic homeostasis in a largely non-thermogenic state. Here, we integrated physiological, histological, transcriptomic, and molecular analyses of back-fat and tail-fat depots during autumn fattening and winter sleep. Despite reduced food intake, body weight loss, and mild hypothermia, raccoon dogs maintained systemic glucose and lipid homeostasis. Both WAT depots exhibited adipocyte atrophy and the coordinated suppression of core metabolic and biosynthetic pathways, indicating a shared program of metabolic depression. However, the two depots adopted distinct remodeling strategies. Back-fat showed collagen densification and vascular-associated remodeling, suggesting a structural adaptation that may preserve tissue integrity during winter sleep. In contrast, tail-fat displayed enhanced innate immune signaling and M2 macrophage enrichment, indicating immune niche remodeling that may support tissue protection during prolonged lipid mobilization. Together, these findings reveal that raccoon dogs maintain metabolic homeostasis during shallow hibernation through a non-thermogenic, WAT-centered strategy characterized by shared metabolic depression and depot-specific structural and immunometabolic remodeling. Full article

(This article belongs to the Section Molecular Endocrinology and Metabolism)

► Show Figures

Figure 1

5 pages, 869 KB

Open AccessEditorial

Special Issue “Fundamental and Practical Perspectives in Regenerative Medicine: Proceedings of the VI National Congress of Regenerative Medicine (2024)”

by Pavel I. Makarevich and Vsevolod A. Tkachuk

Int. J. Mol. Sci. 2026, 27(12), 5610; https://doi.org/10.3390/ijms27125610 - 22 Jun 2026

Viewed by 306

Abstract

The recent rapid advances in technological pipelines are urging faster and riskier translational moves from research to practice in regenerative medicine [...] Full article

(This article belongs to the Special Issue Fundamental and Practical Perspectives in Regenerative Medicine: Proceedings of the VI National Congress of Regenerative Medicine (2024))

► Show Figures

Figure 1

Journal Menu

Journal Browser

Int. J. Mol. Sci., Volume 27, Issue 12 (June-2 2026) – 490 articles

Further Information

Guidelines

MDPI Initiatives

Follow MDPI