International Journal of Molecular Sciences

30 pages, 654 KB

Open AccessReview

Beyond the Needle: Is Liquid Biopsy the Future of Veterinary Medicine?

by Iga Horodyska, Patrycja Kasperska, Daria Będkowska, Sara Al-Ameri, Aleksandra Adam, Izabela Herman, Marta Miszczak and Joanna Bubak

Int. J. Mol. Sci. 2026, 27(18), 8183; https://doi.org/10.3390/ijms27188183 - 14 Sep 2026

Abstract

The detection of circulating tumor DNA (ctDNA) and circulating tumor cells (CTCs) is a minimally invasive approach for diagnosing and monitoring cancer. These liquid biopsy-based strategies enable the identification of primary or metastatic tumors and provide valuable information on tumor biology and treatment [...] Read more.

The detection of circulating tumor DNA (ctDNA) and circulating tumor cells (CTCs) is a minimally invasive approach for diagnosing and monitoring cancer. These liquid biopsy-based strategies enable the identification of primary or metastatic tumors and provide valuable information on tumor biology and treatment response. A variety of techniques are employed to analyze components obtained through liquid biopsy. While CTCs are typically detected using cell-based methods, ctDNA is commonly analyzed using highly sensitive molecular approaches, including quantitative PCR (qPCR), digital PCR (dPCR), and next-generation sequencing (NGS). Extracellular vesicles (EVs), which carry tumor-derived nucleic acids, proteins, and other biomolecules, are increasingly investigated as potential biomarkers, while tumor-educated platelets (TEPs) can reflect tumor-associated molecular changes and may provide additional information for cancer detection and monitoring. Furthermore, emerging multi-cancer early detection (MCED) approaches aim to identify molecular signatures associated with multiple cancer types from a single blood sample, highlighting the broader diagnostic potential of liquid biopsy. Due to the high specificity of tumors and somatic mutations, ctDNA serves as a real-time biomarker for tracking cancer progression. The advantages of ctDNA diagnostics include its low invasiveness and its capacity to detect cancer at early stages. In addition to confirming the presence of a tumor, liquid biopsy approaches facilitate the assessment of malignancy and the evaluation of treatment response. In the field of human medicine, ctDNA plays a pivotal role in diagnostic procedures. It is utilized not only for screening tests that detect the presence of cancer but also for the development of targeted treatment protocols for specific patients. These protocols are informed by the detection of genomic alterations and are designed to monitor the response to therapy over the course of treatment. Similarly, CTC, EV, TEP, and MCED approaches are being investigated as complementary tools for cancer detection, molecular characterization, prognosis, and longitudinal disease monitoring. In the domain of veterinary medicine, ctDNA has been instrumental in the diagnosis of various neoplasms, including osteosarcomas, hemangiosarcomas, lymphomas, canine mammary tumors, and melanomas. Other liquid biopsy components, including CTCs and EVs, also show promise for the detection and characterization of tumors in companion animals, although their clinical application remains less developed than in human medicine. Another significant application is in the detection of minimal residual disease (MRD), where a small number of cancer cells remain undetectable by conventional tests, such as blood counts. This underscores the significance of advanced ctDNA analysis. However, challenges persist, including the low concentration of ctDNA in blood, the low ratio of mutated to normal DNA fragments, potential contamination, biological variability, and the need for standardized protocols. This review synthesizes the current knowledge on liquid biopsy, including ctDNA, CTCs, EVs, TEPs, and emerging MCED approaches, and the potential for transferring these technologies from human medicine to animal medicine. Continued research is necessary to enhance the sensitivity and specificity of detection, which could facilitate early cancer diagnosis in animals and improve survival through timely treatment. Full article

(This article belongs to the Section Molecular Oncology)

20 pages, 3380 KB

Open AccessArticle

Fail-Closed Validation of Lineage-Associated Transcript-Count Diversity After IFN-β Stimulation in a Public Human PBMC Dataset

by Roberto Navarro Quiroz, Katherine Escorcia Lindo, Andrea Jaruffe Pinilla, Yirys Díaz-Olmos, Cecilia Fernández-Ponce, Eloina Zarate Peñata, Yesit Bello Lemus, Lisandro Pacheco Lugo, Leonardo Pacheco Londoño, Antonio Acosta Hoyos, Nataly Galan Freyle, Katy Elena Retamoza Chamorro, Jose Luis Villarreal-Camacho and Elkin Navarro Quiroz

Int. J. Mol. Sci. 2026, 27(18), 8182; https://doi.org/10.3390/ijms27188182 - 14 Sep 2026

Abstract

Shannon entropy of single-cell transcript counts reflects composition and sampling. We tested whether interferon-beta (IFN-β) stimulation was associated with donor-consistent diversity changes after employing technical controls. We reanalyzed GSE96583 peripheral blood mononuclear cells from eight donors with lupus, treating donors—not the 8949 target [...] Read more.

Shannon entropy of single-cell transcript counts reflects composition and sampling. We tested whether interferon-beta (IFN-β) stimulation was associated with donor-consistent diversity changes after employing technical controls. We reanalyzed GSE96583 peripheral blood mononuclear cells from eight donors with lupus, treating donors—not the 8949 target cells obtained from them—as inferential units. Analyses included sampling without replacement at 562 unique molecular identifiers, a 750-molecule sensitivity analysis, Miller–Madow correction, IFN-gene removal, and exact compositional decomposition. Mean paired-donor raw-count plug-in entropy increased by 0.138 bits in CD14+ monocytes and 0.329 bits in FCGR3A+ monocytes and decreased by 0.080 bits in natural killer cells. Directions persisted across prespecified robustness variants. Monocyte increases accompanied higher evenness and lower transcript dominance; natural killer cells showed the opposite pattern, without consistent richness changes. Adding IFN activity to donor-, condition-, and technical-covariate-adjusted models increased R² by at most 0.0108. Entropy contributions strongly overlapped pseudobulk expression changes. In GSE194122, the implemented binary chromatin-accessibility entropy was determined by open-peak count, precluding biological cross-modality coupling claims. These findings support a lineage-divergent association within this single lupus cohort; independent replication is required before broader biological or diagnostic interpretation. Full article

(This article belongs to the Section Molecular Biophysics)

14 pages, 1853 KB

Open AccessArticle

Terminal V5 Tagging of All Four Newcastle Disease Virus Structural Proteins Is Compatible with Particle Formation in Sf9 Cells

by Aurelia Schweda, Dzmitry Dauhalevich and Lukasz Rabalski

Int. J. Mol. Sci. 2026, 27(18), 8181; https://doi.org/10.3390/ijms27188181 - 14 Sep 2026

Abstract

Newcastle disease virus-like particles (NDVLPs) assemble from the matrix (M), nucleoprotein (NP), fusion (F) and hemagglutinin–neuraminidase (HN) proteins. Terminal epitope tags make these proteins easy to detect but may interfere with assembly, and it is not well-documented how much terminal modification the four [...] Read more.

Newcastle disease virus-like particles (NDVLPs) assemble from the matrix (M), nucleoprotein (NP), fusion (F) and hemagglutinin–neuraminidase (HN) proteins. Terminal epitope tags make these proteins easy to detect but may interfere with assembly, and it is not well-documented how much terminal modification the four proteins tolerate. Eight pFastBac Dual donor plasmids were used to generate recombinant baculoviruses, and twelve NDVLP variants were produced in Sf9 cells and purified by sucrose gradient ultracentrifugation. Particle-like structures were recovered from all twelve preparations. Western blotting confirmed production of the native and V5-tagged proteins and their co-purification in the particle fractions; transmission electron microscopy showed enveloped, roughly spherical particles with measured diameters of 55 to 150 nm (median 90 nm) in every preparation, similar in appearance to native NDV. Particles of a similar size were also present in the beta-glucuronidase control preparation, so the assignment of the particles rests on the protein composition of the fractions rather than on morphology alone. Particles formed regardless of which structural protein carried the tag, including a variant in which all four proteins were tagged simultaneously. Under the conditions tested, 19-residue terminal insertions were compatible with NDVLP recovery at each of the four positions examined. Full article

(This article belongs to the Special Issue Viruses as Therapeutic Tools: Medical and Biotechnological Applications)

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14 pages, 5079 KB

Open AccessArticle

OsBTBZ1 Regulates Nitrate Transport in Arabidopsis thaliana via NRT2.1 Modulation Under Salt Stress

by Laurent Soulard, Nopphakhun Khunpolwattana, Chutarat Punchkhon, Triono Bagus Saputro, Luca Comai, Teerapong Buaboocha and Supachitra Chadchawan

Int. J. Mol. Sci. 2026, 27(18), 8180; https://doi.org/10.3390/ijms27188180 - 14 Sep 2026

Abstract

Nitrogen availability and salinity are two major environmental factors affecting plant growth and development. Nitrate uptake is strictly regulated through transcriptional networks that balance nutrient assimilation and stress responses. The role of BTB-ZF transcription factors, which have been implicated in various abiotic stress [...] Read more.

Nitrogen availability and salinity are two major environmental factors affecting plant growth and development. Nitrate uptake is strictly regulated through transcriptional networks that balance nutrient assimilation and stress responses. The role of BTB-ZF transcription factors, which have been implicated in various abiotic stress responses, in coordinating nitrate uptake remains largely unknown; specifically, the precise molecular mechanisms through which OsBTBZ1, a BTB-ZF transcription factor from Oryza sativa, confers salt tolerance remain to be fully elucidated. Here, we characterize the function of OsBTBZ1 in Arabidopsis thaliana under nitrate and salt stress conditions. Transcriptomic analysis of the bt3 mutant and revertant lines expressing OsBTBZ1 in the bt3 mutant background indicated a relationship with NRT2.1, which encodes a high-affinity nitrate transporter. The bt3 mutant exhibited severe root growth inhibition under low nitrate and salt stress; however, expression of OsBTBZ1 restored root growth under combined stress, supporting a role for AtBT3 in stress adaptation. Our findings suggest that OsBTBZ1 may contribute to the coordination of nitrate-responsive pathways and root plasticity under stress, potentially through regulation of nitrate transport-related processes, including those associated with NRT2.1. These findings highlight the potential of OsBTBZ1-mediated regulatory mechanisms for improving nitrogen use efficiency and stress tolerance in future crops. Full article

(This article belongs to the Special Issue Exploring Abiotic Stress in Plants: Mechanisms, Adaptations, and Mitigation Strategies (2nd Edition))

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17 pages, 2055 KB

Open AccessArticle

The First Genomic Study in a Romanian Spotted Calf with Ectopia Cordis Thoracalis

by Viorel Herman, Simona Marc, Oana Maria Boldura, Alina Cărunta, Anca-Alexandra Tămaș, Ioan Claudiu Crăciun, Adrian Olariu-Jurca, Gabriel Otavă, Corina Badea and Alexandru Eugeniu Mizeranschi

Int. J. Mol. Sci. 2026, 27(18), 8179; https://doi.org/10.3390/ijms27188179 - 14 Sep 2026

Abstract

Congenital heart defects (CHDs) are defects present at birth that can often lead to perinatal death, or, rarely, can be asymptomatic and detected later in life. Ectopia cordis is a rare type of malformation in which the heart is not located in its [...] Read more.

Congenital heart defects (CHDs) are defects present at birth that can often lead to perinatal death, or, rarely, can be asymptomatic and detected later in life. Ectopia cordis is a rare type of malformation in which the heart is not located in its normal position. Other congenital abnormalities can be associated with this condition. In this article, we describe a case of ectopia cordis thoracalis with atrial septal defect in association with unilateral renal agenesis in a male Romanian Spotted calf. Clinical examination, dissection, and whole-genome sequencing were performed to describe and identify a possible cause of the abnormalities. Clinical examination revealed the presence of the heart in the thoracic region, subcutaneously, without apparent impairment of major physiological functions. Upon dissection, the external appearance of the heart was normal. However, the internal examination revealed an atrial septal defect measuring 1 cm in diameter in the proximal third of the interatrial septum. The case presented unilateral agenesis of the left kidney. VEP annotation of the joint callset identified 2270 missense variant annotations predicted as deleterious by SIFT and, separately, 12 high-impact start-loss variants by VEP consequence annotation. The OR51F5C and OR8J17 variants were homozygous in the calf. Analyzing genes involved in cardiac and renal development, the following genes—SHH, TBX18, NOTCH1, GATA3, TBX2, FOXC1, and RET had mutations with moderate impact. Mutations in the FOXC1 gene occurred in homozygous states for both the calf and mother genomes, while the RET gene had two mutations in a homozygous state just in the calf genome, and in a heterozygous state in the mother’s genome. One limitation of the present study is the unavailability of the paternal genome for comparative analysis; in addition, maternal genomic profiling did not reveal a contribution to the identified variants. Although the specific pathogenic relevance of the variants detected to the observed phenotype cannot be conclusively established, the findings expand the current genomic knowledge of this rare disorder and provide valuable data for future investigations into its genetic basis. Full article

(This article belongs to the Special Issue Molecular Progression of Genetics in Breeding of Farm Animals, 2nd Edition)

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3 pages, 170 KB

Open AccessComment

The Potential Role of Organ-to-Liver Ratios in Immune-Related FDG PET Imaging. Comment on Panikar et al. Exploratory FDG PET/CT Imaging in mCRPC Patients Treated with Sipuleucel-T ± IL-7: A Phase II Trial Subanalysis. Int. J. Mol. Sci. 2026, 27, 5129

by Alev Çınar

Int. J. Mol. Sci. 2026, 27(18), 8178; https://doi.org/10.3390/ijms27188178 - 14 Sep 2026

Abstract

We read with great interest the article by Panikar et al [...] Full article

(This article belongs to the Topic New Advancements in Innate Immunity and Cancer Immunotherapy)

2 pages, 153 KB

Open AccessCorrection

Correction: Ti et al. Blood-Based α-Synuclein Biomarkers in Parkinson’s Disease: Molecular Diversity, Analytical Advances and Clinical Translation. Int. J. Mol. Sci. 2026, 27, 6254

by Kailiang Ti, Yi Zhao, Eng King Tan and Dongrui Ma

Int. J. Mol. Sci. 2026, 27(18), 8177; https://doi.org/10.3390/ijms27188177 - 14 Sep 2026

Abstract

In the original publication [...] Full article

(This article belongs to the Section Molecular Neurobiology)

17 pages, 5338 KB

Open AccessArticle

Mating System and Fine-Scale Spatial Genetic Structure of the Tropical Epiphytic Orchid Rhynchostylis gigantea

by Wen-Chang Li, Zhi-Heng Chen, Hao-Tian Zhong, Ming-Xun Ren, Zhe Zhang and Xi-Qiang Song

Int. J. Mol. Sci. 2026, 27(18), 8176; https://doi.org/10.3390/ijms27188176 - 14 Sep 2026

Abstract

Habitat fragmentation caused by human activities threatens plant genetic diversity, but the mechanisms shaping gene flow and spatial genetic structure in epiphytic orchids remain poorly understood. Here, we investigated the genetic structure and gene flow patterns of the epiphytic orchid Rhynchostylis gigantea in [...] Read more.

Habitat fragmentation caused by human activities threatens plant genetic diversity, but the mechanisms shaping gene flow and spatial genetic structure in epiphytic orchids remain poorly understood. Here, we investigated the genetic structure and gene flow patterns of the epiphytic orchid Rhynchostylis gigantea in a human-modified landscape on Hainan Island, China, using SNP markers. Based on 2005 high-quality SNPs generated via double-digest restriction site-associated DNA sequencing (ddRADseq) from 275 individuals, we assessed genetic diversity, population differentiation, and fine-scale spatial genetic structure (FSGS) and further explored mating patterns through parentage analysis of 100 F1 offspring. The adult population maintained moderate genetic diversity (H_o = 0.243, F_IS = 0.082), whereas offspring cohorts showed stronger heterozygote deficiency (F_IS = 0.198), suggesting that high contemporary geitonogamous selfing may be counterbalanced by post-zygotic selective mortality during early life stages prior to adult recruitment. The two adult subpopulations separated by agricultural fields exhibited significant genetic differentiation (Φ_PT = 0.131, p = 0.001), indicating reduced gene exchange connectivity caused by landscape fragmentation. Significant FSGS was detected across the population and subpopulations, with three-dimensional spatial analyses revealing the influence of both spatial distance and host-tree distribution. Parentage analysis revealed a high rate of geitonogamy and a pattern in which single pollen donors fertilized multiple flowers on the same recipient plant. This indicates that pollinator movement is highly restricted, leading to localized mating within immediate flower clusters. Our results demonstrate that the genetic structure of R. gigantea is shaped by the combined effects of mixed mating systems, limited seed dispersal, host-tree dependence, and habitat fragmentation. Conservation of epiphytic orchids should therefore integrate the protection of orchid populations, host trees, and landscape connectivity to maintain long-term genetic diversity. Full article

(This article belongs to the Section Molecular Plant Sciences)

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20 pages, 8308 KB

Open AccessArticle

The Role of Glucose Metabolism in Xenopus Primitive Myeloid Cell Development and Function

by Loucas Demetriou, Paris A. Skourides and Neophytos Christodoulou

Int. J. Mol. Sci. 2026, 27(18), 8175; https://doi.org/10.3390/ijms27188175 - 14 Sep 2026

Abstract

Primitive Myeloid Cells (PMCs) arise from the anterior ventral blood islands during Xenopus laevis embryogenesis and migrate to colonize embryonic tissues, contributing to wound healing, innate immunity, and development. Because cell movement and growth are metabolically demanding, we investigated whether metabolic pathways regulate [...] Read more.

Primitive Myeloid Cells (PMCs) arise from the anterior ventral blood islands during Xenopus laevis embryogenesis and migrate to colonize embryonic tissues, contributing to wound healing, innate immunity, and development. Because cell movement and growth are metabolically demanding, we investigated whether metabolic pathways regulate PMC behaviour integrating public single-cell RNA-seq data with whole-mount in situ hybridization (WMISH) to profile metabolic gene expression in PMCs, followed by functional assays using pathway-specific inhibitors: 2-deoxy-D-glucose 2-DG (global glucose metabolism), YZ-9 (glycolysis), 6-AN (pentose phosphate pathway, PPP), and ST045849 (hexosamine biosynthetic pathway, HBP). The analyses showed that PMCs express high levels of glycolytic and ancillary metabolic enzymes. Global inhibition impaired PMC colonization and wound recruitment and modestly reduced the size of the specification domain, confirming a requirement for glycolytic flux. PPP and downstream glycolysis inhibition produced only modest, non-significant shifts in recruitment, whereas HBP inhibition abolished wound recruitment and reduced PMC numbers. These findings identify glucose metabolism as a regulator of PMC colonization and immune function, revealing how metabolic reprogramming governs early immune cell behaviour independent of a functional vasculature. Full article

(This article belongs to the Special Issue Developmental Biology: Computational and Experimental Approaches, 3rd Edition)

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20 pages, 13064 KB

Open AccessArticle

Photocatalytic Reduction of Cr(VI) by MOF@Azo-COF Under Strongly Acidic Conditions

by Xiaolong Gong, Guojian Ren, Qi Zhou, Linglong Yi, Qingxiang Wu, Yingjie Hua and Qinhe Pan

Int. J. Mol. Sci. 2026, 27(18), 8174; https://doi.org/10.3390/ijms27188174 - 14 Sep 2026

Abstract

Cr(VI) poses a serious threat to the ecosystem and human health; in order to efficiently remove Cr(VI), a stable MOF@COF core-shell photocatalyst (MOF-808@TpAzo) was developed. Through a post-modification strategy, MOF-808 was functionalized with amino groups on its surface. Subsequently, the TpAzo-based covalent organic [...] Read more.

Cr(VI) poses a serious threat to the ecosystem and human health; in order to efficiently remove Cr(VI), a stable MOF@COF core-shell photocatalyst (MOF-808@TpAzo) was developed. Through a post-modification strategy, MOF-808 was functionalized with amino groups on its surface. Subsequently, the TpAzo-based covalent organic framework (COF) was grown in-situ on its surface, successfully yielding the core-shell structured MOF-808@TpAzo composite. Under highly acidic conditions, the MOF-808@TpAzo composite shows an excellent photocatalytic reduction of Cr(VI) performance, which is significantly better than that of its single component. Especially, as little as 5 mg of the catalyst, 99.42% of Cr(VI), was removed from the acidic simulated wastewater within 120 min, and the material exhibited excellent cycling stability. The highly effective catalysis is attributed to the heterojunction formed via the covalent connection between the MOF and COF components, promoting charge separation and transport. This work provides a new alternative for developing stable photocatalysts capable of the reduction of Cr(VI) in strongly acidic environments. Full article

(This article belongs to the Special Issue The Design, Synthesis, and Application of Functional Porous Materials)

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28 pages, 413 KB

Open AccessReview

Macrophage–Fibroblast Crosstalk in Kidney Injury: A Narrative Review

by Panpan Qiang, Lizheng Han, Ruyan Lv, Zhenfei Dong, Teng Ma, Xiangting Wang and Qingyou Xu

Int. J. Mol. Sci. 2026, 27(18), 8173; https://doi.org/10.3390/ijms27188173 - 14 Sep 2026

Abstract

Renal fibrosis is a multifaceted pathological process driven by cellular interactions, with fibroblasts and macrophages serving as central regulators. Fibroblasts activate into matrix-producing myofibroblasts, directly contributing to fibrogenesis and disease progression. Macrophages, characterized by pronounced heterogeneity, dynamically modulate the balance between tissue injury [...] Read more.

Renal fibrosis is a multifaceted pathological process driven by cellular interactions, with fibroblasts and macrophages serving as central regulators. Fibroblasts activate into matrix-producing myofibroblasts, directly contributing to fibrogenesis and disease progression. Macrophages, characterized by pronounced heterogeneity, dynamically modulate the balance between tissue injury and repair, thereby critically influencing disease outcomes. The crosstalk between these two cell types operates through diverse signaling pathways, extracellular vesicle-mediated communication, and metabolic reprogramming, forming an intricate regulatory network that exerts stage-specific effects during renal injury. Recent insights into fibroblast activation, macrophage polarization, and their intercellular interplay have provided potential therapeutic avenues for delaying kidney fibrosis, offering renewed promise for clinical translation. Full article

(This article belongs to the Special Issue The Role of Fibroblasts in the Regulation of Tissue Inflammation and Repair)

28 pages, 2775 KB

Open AccessArticle

Snowmaking Infrastructure: A Selective Barrier or Potential Vector for Antibiotic Residues and Resistance Determinants in Mountain Catchments

by Natalia Czernecka-Borchowiec, Klaudia Stankiewicz, Klaudia Bulanda, Piotr Boroń, Justyna Prajsnar, Anna Kopacz, Zofia Schejbal and Anna Lenart-Boroń

Int. J. Mol. Sci. 2026, 27(18), 8172; https://doi.org/10.3390/ijms27188172 - 14 Sep 2026

Abstract

Snowmaking systems are increasingly used to serve winter tourism in mountain regions, yet their role in the environmental dissemination of antimicrobial resistance (AMR) remains poorly understood. Here, we investigated how snowmaking infrastructure affects the transport, retention and reduction of antibiotic residues, antibiotic resistance [...] Read more.

Snowmaking systems are increasingly used to serve winter tourism in mountain regions, yet their role in the environmental dissemination of antimicrobial resistance (AMR) remains poorly understood. Here, we investigated how snowmaking infrastructure affects the transport, retention and reduction of antibiotic residues, antibiotic resistance genes (ARGs) and phenotypically resistant bacteria along two interconnected water chains feeding technical snow production in mountain catchments. Over two winter seasons (2023/24 and 2024/25), we quantified a panel of clinically relevant antibiotics, ARGs and resistance phenotypes in wastewater effluent, intake water, reservoirs, technical snow, aged snow, meltwater and receiving streams; culturable bacteria, resistance phenotypes and ARGs were additionally assessed in reservoir sediments and snow cannon filter material. Wastewater and intake water exhibited the highest antibiotic loads and ARG richness, dominated by fluoroquinolones, macrolides and β-lactam resistance determinants. Reservoirs accumulated substantial amounts of antibiotics but their reduction occurred further along the snowmaking chain. Fresh technical snow still contained multiple antibiotics and ARGs, and harbored resistant Enterobacteriaceae, indicating that snowmaking systems can redistribute AMR determinants into the snowpack. Aged snow, meltwater and receiving streams showed nearly complete loss of antibiotic residues and reduced ARG richness, due to processes of snow transformation and downstream transport. By integrating chemical, molecular and phenotypic data with the technical layout of the infrastructure, we identify specific components as AMR retention hotspots and clarify where barrier functions are most effective. Our findings demonstrate that snowmaking infrastructure can both concentrate and distribute antibiotic residues and resistance determinants in mountain catchments, highlighting the need to consider snowmaking systems in environmental AMR risk assessments and One Health frameworks. Full article

(This article belongs to the Special Issue Molecular Research on Micropollutants in Various Enviroments)

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40 pages, 13560 KB

Open AccessArticle

A2BAR Antagonism Mitigates Pulmonary Inflammation and Fibrosis and Restores Macrophage M1/M2 Balance in Diabetic Rats

by Ángelo Torres-Arévalo, Sarah Montoya-Muñoz, Mateo Montes-Vanegas, Claudio Cappelli-León, Ignacio Árias, Claudia Jara Cancino, Claudia Quezada, Sebastián Alarcón, Benjamín Guerrero, Pamela Silva, Víctor Pola-Véliz and Rody San Martín

Int. J. Mol. Sci. 2026, 27(18), 8171; https://doi.org/10.3390/ijms27188171 - 14 Sep 2026

Abstract

The pathophysiological mechanisms underlying diabetic lung disease (DLD) remain poorly understood. Beyond hyperglycemia, dysregulated molecules such as adenosine may contribute to pulmonary inflammation and fibrosis. MRS1754, a selective antagonist of the A2B adenosine receptor (A2BAR), has demonstrated anti-inflammatory and antifibrotic effects, including reductions [...] Read more.

The pathophysiological mechanisms underlying diabetic lung disease (DLD) remain poorly understood. Beyond hyperglycemia, dysregulated molecules such as adenosine may contribute to pulmonary inflammation and fibrosis. MRS1754, a selective antagonist of the A2B adenosine receptor (A2BAR), has demonstrated anti-inflammatory and antifibrotic effects, including reductions in macrophage (MΦ) infiltration in animal models of diabetes-associated complications; however, its effects on DLD remain unexplored. Here, diabetic Sprague-Dawley rats were treated with MRS1754, and pulmonary inflammation, fibrosis, MΦ infiltration, and polarization were assessed in bronchoalveolar lavage fluid (BALF) and lung tissue using immunohistopathological and molecular approaches. According to data normality, one-way ANOVA/Tukey’s test or Kruskal–Wallis/Dunn’s test were employed, as appropriate; ordinal data were analyzed using the Mann–Whitney test with Bonferroni correction. Compared with controls, diabetic rats exhibited pulmonary inflammation and fibrosis, increased MΦ infiltration, and an M1/M2 polarization imbalance. MRS1754 attenuated these alterations, reducing Tumor necrosis factor-alpha (TNF-α) and Transforming growth factor-beta 1 (TGF-β1) levels, decreasing MΦ accumulation in BALF and lung tissue, and fully restoring the M1/M2 balance to control levels. BALF MΦ populations, particularly the M2 (Cluster of differentiation 163-positive; CD163⁺) subset, correlated strongly with TGF-β1 and fibronectin (FN1) levels. Reanalysis of human fibrotic lung single-cell transcriptomic data further identified MΦ as a major TGF-β1-expressing cell population. Collectively, these findings suggest that A2BAR antagonism partially mitigates inflammation, fibrosis, and MΦ infiltration while fully restoring MΦ polarization balance in DLD, supporting its potential as a therapeutic strategy. Full article

(This article belongs to the Special Issue Mechanisms and Treatments of Adenosine Signaling-Regulated Diseases)

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16 pages, 8807 KB

Open AccessReview

Extracellular Hemoglobin, Hypoxia, and Macrophage-Mediated Pulmonary Vascular Remodeling in Hemolytic Disease

by Melissa J. Lucero, Eva Nozik, Kathryn Hassell, David C. Irwin, Paul W. Buehler and Scott K. Ferguson

Int. J. Mol. Sci. 2026, 27(18), 8170; https://doi.org/10.3390/ijms27188170 - 14 Sep 2026

Abstract

Pulmonary hypertension (PH) is a well-recognized complication of chronic hemolytic anemias such as sickle cell disease and thalassemia, yet the relative contributions of hypoxia and cell-free hemoglobin (Hb) to disease progression remain incompletely understood. Patients with hemolytic anemia experience a lifelong cycle of [...] Read more.

Pulmonary hypertension (PH) is a well-recognized complication of chronic hemolytic anemias such as sickle cell disease and thalassemia, yet the relative contributions of hypoxia and cell-free hemoglobin (Hb) to disease progression remain incompletely understood. Patients with hemolytic anemia experience a lifelong cycle of chronic and inter bitten hypoxia that compounds vascular injury driven by extracellular Hb and its degradation products, heme and iron. While the effects of hypoxia and Hb exposure have historically been studied in isolation, the combined impact of sustained, low-level plasma Hb together with chronic hypoxia—more representative of steady-state hemolysis—has been largely overlooked. A rat model incorporating chronic hypoxia with continuous low-dose Hb infusion via an implanted pump demonstrates that even modest plasma Hb concentrations (10–20 µM heme) exert an additive effect on hypoxia-induced PH. This effect is associated with increased adventitial macrophage accumulation, oxidative stress, and inflammation, driving more severe pulmonary vascular remodeling. Building on this model, therapeutic strategies targeting Hb-mediated vascular injury are evaluated, with particular focus on repeated-dose haptoglobin (Hp) therapy, given that Hp is often severely depleted in sickle cell disease. Restoring circulating Hp sequesters plasma Hb into a non-reactive, compartmentalized Hb–Hp complex, limiting NO scavenging and oxidative damage. These mechanistic findings are further linked to functional outcomes through studies of skeletal muscle microvascular oxygen tension and exercise capacity in Berkeley sickle cell disease mice. This review synthesizes findings across these studies to clarify the interplay between hypoxia, macrophage biology, and extracellular Hb in driving pulmonary vascular remodeling and to highlight emerging Hb-targeted therapeutic strategies for hemolysis-associated PH. Full article

(This article belongs to the Special Issue Advances in Cardiovascular and Vascular Biology)

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26 pages, 39638 KB

Open AccessArticle

Ligusticum sinense Oliv. cv. Chaxiong Extract Alleviates Nitroglycerin-Induced Migraine-like Responses in Rats: Associations with Glycerophospholipid Metabolism and PI3K/AKT Signaling

by Yu Zhu, Yanjing Dong, Shiyi Jiang, Qian Qin, Yu He, Danyang Wu, Shouwen Zhang and Juan Wei

Int. J. Mol. Sci. 2026, 27(18), 8169; https://doi.org/10.3390/ijms27188169 - 14 Sep 2026

Abstract

Ligusticum sinense Oliv. cv. Chaxiong (Chaxiong) has traditionally been used as a medicinal tea for headache relief in southern China, but its bioactive basis and mechanisms remain unclear. This study integrated ultra-performance liquid chromatography–quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS), untargeted metabolomics, network pharmacology, [...] Read more.

Ligusticum sinense Oliv. cv. Chaxiong (Chaxiong) has traditionally been used as a medicinal tea for headache relief in southern China, but its bioactive basis and mechanisms remain unclear. This study integrated ultra-performance liquid chromatography–quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS), untargeted metabolomics, network pharmacology, molecular docking, reverse-transcription quantitative PCR (RT-qPCR), and Western blotting in a nitroglycerin-induced migraine-like rat model. Chaxiong extract alleviated migraine-like behavior, reduced calcitonin gene-related peptide (CGRP), nitric oxide (NO), and tumor necrosis factor-α (TNF-α), increased 5-hydroxytryptamine (5-HT), and ameliorated brain histopathological alterations. A total of 48 constituents and 13 differential serum metabolites were annotated or putatively annotated, respectively. Glycerophospholipid metabolism was the principal metabolic pathway affected by treatment, whereas network pharmacology prioritized phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) signaling. Pathway integration identified PIK3CA, PIK3CB, AKT1, AKT2, and INS as candidate bridging targets. Exploratory docking suggested potential interactions between five representative constituents and PIK3CA/AKT1. RT-qPCR showed reduced Pik3ca and Akt1 mRNA expression, while Western blotting showed decreased p-PI3K p85α (Tyr607)/PI3K p85α and p-AKT (Ser473)/AKT1 ratios following treatment. These findings suggest that Chaxiong-associated alleviation of migraine-like responses was accompanied by altered glycerophospholipid metabolism and reduced PI3K/AKT pathway activation. Full article

(This article belongs to the Section Bioactives and Nutraceuticals)

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18 pages, 4411 KB

Open AccessArticle

Peripheral Blood Transcriptomic Profile Associated with Cutaneous Radiation Injury in Cancer Patients Undergoing Radiotherapy

by Seid Muhie, Nabarun Chakraborty, Mital Patel, Alison Ross, Lauren Moffatt, Melissa McLawhorn, Aarti Gautam, Jeffrey Shupp and Rasha Hammamieh

Int. J. Mol. Sci. 2026, 27(18), 8168; https://doi.org/10.3390/ijms27188168 - 14 Sep 2026

Abstract

The peripheral blood transcriptomic profile associated with clinically observed cutaneous radiation injury (CRI) during fractionated radiotherapy remains incompletely characterized. We analyzed blood transcriptomes from 22 adults receiving radiotherapy: 10 with CRI and 12 without radiation-related skin injury at blood collection. RNA was assayed [...] Read more.

The peripheral blood transcriptomic profile associated with clinically observed cutaneous radiation injury (CRI) during fractionated radiotherapy remains incompletely characterized. We analyzed blood transcriptomes from 22 adults receiving radiotherapy: 10 with CRI and 12 without radiation-related skin injury at blood collection. RNA was assayed using two-color Agilent microarrays. Variance filtering retained 12,251 probes that were evaluated using four complementary analyses: preranked Gene-Set Enrichment Analysis (GSEA) for ranked gene-set enrichment, Gene-Set Variation Analysis (GSVA) for sample-level gene-set scoring, weighted gene co-expression network analysis for co-expression structure, and differential expression analysis at the probe level. GSEA identified positive enrichment of interferon-γ response and E2F targets and negative enrichment of heme metabolism. Consistently, GSVA showed higher interferon-response, E2F-target, unfolded-protein-response, and cholesterol-homeostasis scores and lower angiogenesis, xenobiotic-metabolism, protein-secretion, hypoxia, and heme-metabolism scores in participants with CRI. Ten modular co-expression networks met the data-derived CRI-correlation prioritization criterion. Positively correlated networks were enriched for negative regulation of response to wounding and complement/coagulation cascades. Intramodular connectivity and sample-level expression analyses prioritized KDM4C and PARPBP as exploratory high-connectivity candidates, with both retaining top-10 connectivity in 95.5% of leave-one-out iterations. Exploratory differential expression analysis identified 1842 probes, comprising 585 upregulated and 1257 downregulated probes in CRI. Genes mapped from downregulated probes were significantly enriched for ribosome, spliceosome, and oxidative phosphorylation pathways. Across complementary analytical frameworks, CRI was associated with a coherent peripheral blood profile characterized by interferon and cell-cycle enrichment; altered wound-response and complement/coagulation processes; reduced heme-metabolism, angiogenesis, and xenobiotic-metabolism scores; and downregulated transcripts involved in ribosomal function, RNA processing, and mitochondrial energy metabolism. Although the cross-sectional design and later sampling of CRI participants precluded full separation of CRI-associated differences from treatment-time and cumulative-exposure effects, these findings extend the molecular characterization of CRI and identify pathways and high-connectivity candidates for longitudinal assessment of reproducibility, temporal dynamics, and potential diagnostic or predictive relevance. Full article

(This article belongs to the Special Issue Biological Effects of Radiation on Human Cells and Tissues)

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24 pages, 4701 KB

Open AccessReview

Ion Channel-Targeting Modulators in Glioma: Pharmacological Advances and Therapeutic Perspectives

by Sheng-Nan Wu, Sheng-Che Lin and Rasa Liutkevičienė

Int. J. Mol. Sci. 2026, 27(18), 8167; https://doi.org/10.3390/ijms27188167 - 14 Sep 2026

Abstract

Highly aggressive gliomas remain difficult to treat because of their marked invasiveness, frequent recurrence, and association with glioma-related epilepsy. Increasing evidence suggests that ion channels expressed in glioma cells, including voltage-gated Na⁺ channels, large-conductance Ca²⁺-activated K⁺ (BK_Ca) [...] Read more.

Highly aggressive gliomas remain difficult to treat because of their marked invasiveness, frequent recurrence, and association with glioma-related epilepsy. Increasing evidence suggests that ion channels expressed in glioma cells, including voltage-gated Na⁺ channels, large-conductance Ca²⁺-activated K⁺ (BK_Ca) channels, intermediate-conductance Ca²⁺-activated K⁺ (IK_Ca) channels, and inwardly rectifying K⁺ (Kir) channels, contribute to these malignant phenotypes and represent potential therapeutic targets. This perspective summarizes current evidence for eight representative drugs and phytoconstituents—perampanel, valproic acid, cilostazol, berberine, oxaliplatin, temozolomide, arecoline, and triptolide—that modulate these ion channels. Perampanel and valproic acid suppress voltage-gated Na⁺ current in glioma cells and may provide additional benefits in controlling glioma-associated epilepsy. Cilostazol activates BK_Ca channels, whereas berberine, oxaliplatin, and temozolomide inhibit IK_Ca channel activity. Arecoline suppresses IK_Ca and Kir channel activity, while triptolide inhibits Kir channels. These observations suggest that ion channel modulation may influence glioma cell proliferation, migration, invasion, and excitability. To complement the available experimental evidence, we performed molecular docking analyses as a hypothesis-generating approach to explore potential ligand–channel interactions. Because these computational predictions have not been experimentally validated, they should be interpreted cautiously and viewed as a framework for future mechanistic studies rather than definitive evidence of binding. We also discuss important translational considerations, including the limited clinical evidence and the toxicity profiles of several compounds. In particular, arecoline is associated with carcinogenicity and neurotoxicity, triptolide with significant systemic toxicity, berberine with poor oral bioavailability and potential drug interactions, and oxaliplatin with dose-limiting neurotoxicity. Overall, this perspective highlights ion channels as promising therapeutic targets in glioma while emphasizing the need for rigorous experimental validation and careful evaluation of safety before clinical translation. Full article

(This article belongs to the Special Issue Ion Regulation in Human Pathophysiology)

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22 pages, 1908 KB

Open AccessArticle

Reassessment of IRES Element Activity in Komagataella phaffii Using a Standardized Bicistronic System: Cryptic Promoter Activity Mimics Cap-Independent Translation

by Danil S. Kalinin, Ekaterina V. Schetinina, Alexey N. Fedorov and Igor E. Granovsky

Int. J. Mol. Sci. 2026, 27(18), 8166; https://doi.org/10.3390/ijms27188166 - 14 Sep 2026

Abstract

Internal ribosome entry site (IRES) elements have been proposed for constructing polycistronic mRNAs in Komagataella phaffii, but their reported activity is inconsistent and has not been evaluated under standardized conditions. Here, a standardized bicistronic EGFP-IRES-mKate2 system was developed in which the cap-dependent [...] Read more.

Internal ribosome entry site (IRES) elements have been proposed for constructing polycistronic mRNAs in Komagataella phaffii, but their reported activity is inconsistent and has not been evaluated under standardized conditions. Here, a standardized bicistronic EGFP-IRES-mKate2 system was developed in which the cap-dependent first cistron (EGFP) reports promoter activity, and translation of the second (mKate2) should depend on the IRES. The RhPV, TEV, PVY, and Saccharomyces cerevisiae GPR1 elements were assessed under P_AOX1 and P_THI11, with EMCV and HCV as negative controls. EGFP tracked promoter induction and repression, whereas mKate2 remained at background for GPR1, EMCV, and HCV; for RhPV, TEV, and PVY it was low, promoter-independent, and persisted after promoter deletion. By RT-qPCR, the transcript was intact across both cistrons, except in the RhPV construct, where its 3′ region was depleted. The residual synthesis therefore reflected cryptic promoter activity below 1% of that of P_AOX1, not IRES-dependent initiation. None of the elements directed detectable cap-independent translation above 0.4% of cap-dependent expression of the same ORF, limiting their use for polycistronic expression. The system, the first to place IRES activity in K. phaffii on a quantitative scale, distinguishes IRES-dependent translation from cryptic transcription and can facilitate the search for functional elements. Full article

(This article belongs to the Collection Advances in Cell and Molecular Biology)

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24 pages, 5120 KB

Open AccessArticle

Microemulsions of Essential Oils: Characterization and Phase Behavior

by Dilek Gazolu-Rusanova, Zlatina Mitrinova, Ivan Lesov and Slavka Tcholakova

Int. J. Mol. Sci. 2026, 27(18), 8165; https://doi.org/10.3390/ijms27188165 - 14 Sep 2026

Abstract

Essential oils are a key ingredient in foods, beverages, and personal- and home-care products due to their flavors, scents, and various biological activities. This study investigates the capacity of polysorbate 60 (PS60) micelles to solubilize eight essential oils classified into two groups based [...] Read more.

Essential oils are a key ingredient in foods, beverages, and personal- and home-care products due to their flavors, scents, and various biological activities. This study investigates the capacity of polysorbate 60 (PS60) micelles to solubilize eight essential oils classified into two groups based on their composition. The first group (tea tree, rose, lavender, and melissa oils) contains more than 40% polar aliphatic or alicyclic alcohols or aldehydes, whereas the oils from the second group (chamomile, yarrow, lemon, and orange oils) contain less than 20% of these compounds. The efficient solubilization of the studied essential oils within PS60 micelles is proposed to occur via a mechanism involving two steps: (1) the formation of mixed micelles between PS60 molecules and the polar alcohol or aldehyde constituents of the essential oils, and (2) the dehydration of PS60 ethoxy groups in the presence of highly water-soluble additives (such as citric acid and sodium citrate), which is suggested to decrease the hydrophilic barrier of the micellar corona, thereby facilitating the mass transfer of hydrophobic oil molecules into the micellar core. Stable, transparent microemulsions form only with the first group of essential oils in a certain concentration range. The second step in the proposed mechanism leads to the unexpected finding that reducing the concentration of water-soluble additives, such as citric acid and sodium citrate, impairs solubilization by allowing the micellar corona to rehydrate. Full article

(This article belongs to the Special Issue Molecular Advances on Solubility and Diffusivity in Water-Based Systems and in Biological Media)

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