International Journal of Molecular Sciences

18 pages, 2298 KiB

Open AccessArticle

Rapid Specific PCR Detection Based on THCAS and CBDAS for the Prediction of Cannabis sativa Chemotypes: Drug, Fiber, and Intermediate

by Patwira Boonjing, Worakorn Wiwatcharakornkul, Chayapol Tungphatthong, Taksina Chuanasa, Somchai Keawwangchai, Tae-Jin Yang, Wanchai De-Eknamkul and Suchada Sukrong

Int. J. Mol. Sci. 2025, 26(11), 5077; https://doi.org/10.3390/ijms26115077 (registering DOI) - 24 May 2025

Abstract

Cannabis sativa L. is divided into three main groups: drug-type, intermediate-type, and fiber-type. The presence of tetrahydrocannabinol (THC) exceeding 0.2–0.3% in drug-type and intermediate Cannabis that utilized for recreational and medicinal purposes renders them illegal due to potential mental health implications. Fiber-type contains [...] Read more.

Cannabis sativa L. is divided into three main groups: drug-type, intermediate-type, and fiber-type. The presence of tetrahydrocannabinol (THC) exceeding 0.2–0.3% in drug-type and intermediate Cannabis that utilized for recreational and medicinal purposes renders them illegal due to potential mental health implications. Fiber-type contains high cannabidiol (CBD) and low THC, making it suitable for household use such as textiles and animal feed. Accurate classification is essential to prevent misuse of the plant. High-performance thin-layer chromatography (HPTLC) and ultra-performance liquid chromatography (UPLC), used respectively for the qualitative and quantitative analyses of THC and CBD particularly in female inflorescences, categorized 85 samples of 46 cultivars used in this study into three distinct chemotypes. While chemotype analysis of a very specific organ of the plants accurately identifies Cannabis groups, it requires time-consuming plant development to maturity. Genotype analysis targeting tetrahydrocannabinolic acid synthase (THCAS) and cannabidiolic acid synthase (CBDAS) genes offers a faster alternative for classifying Cannabis types, allowing for sample determination from any part at any developmental stage of the plant. DNA sequencing allowed a phylogenetic analysis based on these genes, classifying all 85 samples of 46 cultivars into the same three groups identified by chemotype analysis. This study is the first to successfully examine the relationship between chemotype and genotype in 85 samples of 46 cultivars. Rapid identification of Cannabis types through genotype analysis lays the groundwork for future development of detection kits. Full article

(This article belongs to the Special Issue Plant Biodiversity and Molecular Marker Technology: Discovery and Application of DNA Polymorphisms)

17 pages, 860 KiB

Open AccessArticle

Serums miR-24-3p and miR-1301-3p as Potential Biomarkers in MEN1 Syndrome

by Simone Donati, Cinzia Aurilia, Francesca Marini, Francesca Giusti, Gaia Palmini, Irene Falsetti, Federica Cioppi, Francesco Ranaldi, Teresa Iantomasi, Arcangelo Moro, Francesco Tonelli and Maria Luisa Brandi

Int. J. Mol. Sci. 2025, 26(11), 5076; https://doi.org/10.3390/ijms26115076 (registering DOI) - 24 May 2025

Abstract

Multiple endocrine neoplasia type 1 (MEN1) is a rare hereditary tumor syndrome caused by inactivating mutations of the MEN1 gene and characterized by the occurrence of multiple endocrine tumors within a single patient (i.e., parathyroid, pituitary, and pancreatic neuroendocrine tumors (NETs)). However, the [...] Read more.

Multiple endocrine neoplasia type 1 (MEN1) is a rare hereditary tumor syndrome caused by inactivating mutations of the MEN1 gene and characterized by the occurrence of multiple endocrine tumors within a single patient (i.e., parathyroid, pituitary, and pancreatic neuroendocrine tumors (NETs)). However, the lack of a genotype–phenotype correlation does not allow individual disease evolution to be foreseen. Epigenetic factors, such as microRNAs, are suspected to contribute to MEN1 tumorigenesis, presumably explaining the lack of genotype–phenotype association. Our previous studies indicated miR-24-3p, miR-1301-3p, miR-664a-3p, and miR-4258 as potentially involved in MEN1 parathyroid tumorigenesis. In this study, we examined the expression of two circulating microRNAs (c-miRNAs), miR-24-3p and miR-1301-3p, in the serum of MEN1 patients. c-miRNAs were evaluated by RT-qPCR in serum collected from 25 MEN1 patients and 25 age- and gender-matched healthy volunteers (HCs). Receiver operating characteristic (ROC) curves were constructed to determine miRNA sensitivity and specificity. RT-PCR analysis revealed that expression levels of circulating miR-1301-3p were significantly downregulated, while those of miR-24-3p were significantly upregulated in the serum of MEN1 patients compared to HCs. Additionally, ROC analysis exhibited a good diagnostic power for both miRNAs (area under the ROC curve (AUC) values: 0.7356 and 0.7928 for miR-1301-3p and miR-24-3p, respectively) in distinguishing MEN1 patients from matched HCs. These preliminary data suggest circulating miR-1301-3p and miR-24-3p as potential non-invasive diagnostic biomarkers for MEN1 syndrome, regardless of different clinical phenotypes and MEN1 mutation types. Full article

(This article belongs to the Section Molecular Biology)

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18 pages, 3224 KiB

Open AccessArticle

Astilbin Alleviates IL-17-Induced Hyperproliferation and Inflammation in HaCaT Cells via Inhibiting Ferroptosis Through the cGAS-STING Pathway

by Xiaohan Xu, Huizhong Zhang, Aqian Chang, Hulinyue Peng, Shiman Li, Ke Zhang, Wenqi Wang, Xingbin Yin, Changhai Qu, Xiaoxv Dong and Jian Ni

Int. J. Mol. Sci. 2025, 26(11), 5075; https://doi.org/10.3390/ijms26115075 (registering DOI) - 24 May 2025

Abstract

Psoriasis, a chronic inflammatory skin disorder, is driven by dysregulated immune responses and keratinocyte dysfunction. Here, we explore the therapeutic potential of Astilbin (AST), a flavonoid with potent anti-inflammatory properties, in modulating ferroptosis and the cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) [...] Read more.

Psoriasis, a chronic inflammatory skin disorder, is driven by dysregulated immune responses and keratinocyte dysfunction. Here, we explore the therapeutic potential of Astilbin (AST), a flavonoid with potent anti-inflammatory properties, in modulating ferroptosis and the cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) pathway in IL-17-stimulated HaCaT keratinocytes. Our psoriatic cell model recapitulated key pathological features, including hyperproliferation, membrane integrity loss, mitochondrial dysfunction, and heightened oxidative stress, alongside elevated proinflammatory cytokine levels. Ferroptosis-related biomarkers were significantly altered, with increased malondialdehyde (MDA) accumulation, reduced glutathione (GSH) levels, iron overload (Fe²⁺), and enhanced lipid peroxidation (detected via C11-BODIPY). Mechanistically, mitochondrial damage triggered cytoplasmic leakage of mitochondrial DNA (mtDNA), activating the cGAS-STING pathway, as evidenced by upregulated pathway-associated protein expression. AST intervention effectively mitigated these pathological changes by suppressing ferroptosis and modulating cGAS-STING signaling. These findings reveal a dual-pathway regulatory mechanism, positioning AST as a promising therapeutic candidate for psoriasis. By elucidating the interplay between ferroptosis and the cGAS-STING pathway, this study provides new insights into psoriatic inflammation and offers a rationale for targeting these pathways in therapeutic strategies. Full article

(This article belongs to the Section Molecular Pharmacology)

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21 pages, 10284 KiB

Open AccessArticle

Genome-Wide Identification of the PR-1 Gene Family in Pyrus betulaefolia Bunge and Its Expression Analysis Under Fire Blight Stress

by Abudusufuer Wufuerjiang, Jingyi Sai, Yue Wen, Lei Wang, Chen Chen, Xu Li and Jia Tian

Int. J. Mol. Sci. 2025, 26(11), 5074; https://doi.org/10.3390/ijms26115074 (registering DOI) - 24 May 2025

Abstract

Fire blight, caused by Erwinia amylovora, is a devastating bacterial disease threatening apple, pear, and other Rosaceae species. In our prior study, transcriptome analysis identified a fire blight-resistant variety, Duli (Pyrus betulifolia Bunge), and highlighted the PR1 gene as a key [...] Read more.

Fire blight, caused by Erwinia amylovora, is a devastating bacterial disease threatening apple, pear, and other Rosaceae species. In our prior study, transcriptome analysis identified a fire blight-resistant variety, Duli (Pyrus betulifolia Bunge), and highlighted the PR1 gene as a key resistance factor. Using Duli’s genomic data, we systematically identified and characterized the Pb-PR-1 gene family through bioinformatics analysis. A total of 31 Pb-PR-1 genes were found, encoding proteins of 123–341 amino acids. Phylogenetic analysis grouped these genes into four subfamilies, with 27 genes distributed across seven chromosomes, all contain a conserved CAP superfamily domain. Their promoter regions were enriched in hormone and stress-responsive elements. After inoculation with E. amylovora, susceptible Duli showed lesion development by day 2, with rapid disease progression, while resistant plants exhibited slower disease advancement and smaller lesions. Enzyme activity assays revealed that in resistant plants, PPO (polyphenol oxidase) and CAT (catalase) activities peaked on day 6, showing a 2.4-fold and 3.81-fold increase compared to susceptible Duli. At the same time, MDA (malondialdehyde) content decreased by 16.6%. The activities of SOD (superoxide dismutase) and PAL (phenylalanine ammonia-lyase) peaked on day 4, with increments of 34.32% and 47.1% over susceptible Duli. qRT-PCR showed significant differences in Pb-PR-1 gene family expression between resistant and susceptible plants post-inoculation. Notably, Pb-PR-1-11, Pb-PR-1-21, and Pb-PR-1-26 expression increased with infection duration, aligning with PPO and CAT activity trends. Other genes showed high early infection expression but declined by day 6. Pb-PR-1-3, Pb-PR-1-6, Pb-PR-1-8, Pb-PR-1-16, and Pb-PR-1-30 were upregulated 13.17-fold on average by day 2. In summary, the Pb-PR-1 family exhibited elevated expression during early infection and enhanced defense-related enzyme activities, improving plant resistance. This study provides a foundation for understanding the PR-1 family’s role in Duli and advancing fire blight resistance in Pyrus species. Full article

(This article belongs to the Section Molecular Plant Sciences)

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16 pages, 2534 KiB

Open AccessArticle

TGF-β Induced by Allergic Lung Inflammation Enhances Os-Teosarcoma Lung Metastasis in a Mouse Comorbidity Model

by Marco J. Sanchez-Rojas, Belen Tirado-Rodriguez, Gabriela Antonio-Andres, Giovanny Soca-Chafre, Daniel D. Hernandez-Cueto, Cesar O. Martinez-Calderon, Mayra Montecillo-Aguado, Juan C. Hernandez-Guerrero, Marco A. Duran-Padilla, Rogelio Hernandez-Pando and Sara Huerta-Yepez

Int. J. Mol. Sci. 2025, 26(11), 5073; https://doi.org/10.3390/ijms26115073 (registering DOI) - 24 May 2025

Abstract

TGF-β is a central mediator of pulmonary allergic inflammation recently associated with lung metastasis of osteosarcoma. Given the controversial links between cancer and allergic diseases, this study aimed to evaluate the effects of allergic airway inflammation—particularly TGF-β—on osteosarcoma lung metastasis using a comorbidity [...] Read more.

TGF-β is a central mediator of pulmonary allergic inflammation recently associated with lung metastasis of osteosarcoma. Given the controversial links between cancer and allergic diseases, this study aimed to evaluate the effects of allergic airway inflammation—particularly TGF-β—on osteosarcoma lung metastasis using a comorbidity mouse model. Osteosarcoma cells were implanted in BALB/c mice with induced allergic airway inflammation. Lung metastasis was quantified, while PCNA/TGF-β expression was assessed by immunohistochemistry and digital pathology. Bioinformatic analyses of patient datasets compared TGF-β and PCNA expression in metastatic vs. normal tissues, and their association with survival. Mice with allergic inflammation showed increased lung metastases associated with TGF-β production. In patient samples, both TGF-β and PCNA were upregulated in metastatic tissues and correlated with poor overall survival. PCNA was also linked to genes involved in cell proliferation, DNA replication, and repair. Our results show an association between allergic airway inflammation and extensive lung metastasis of osteosarcoma in a comorbidity mouse model with elevated expression of TGF-β and PCNA. Full article

(This article belongs to the Special Issue Molecular Perspectives in Lung Diseases: Pathogenesis, Diagnosis, and Treatment)

16 pages, 2634 KiB

Open AccessArticle

QTL Mapping and Developing KASP Markers for High-Temperature Adult-Plant Resistance to Stripe Rust in Argentinian Spring Wheat William Som (PI 184597)

by Arjun Upadhaya, Meinan Wang, Chao Xiang, Nosheen Fatima, Sheri Rynearson, Travis Ruff, Deven R. See, Michael Pumphrey and Xianming Chen

Int. J. Mol. Sci. 2025, 26(11), 5072; https://doi.org/10.3390/ijms26115072 (registering DOI) - 24 May 2025

Abstract

Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is a destructive disease of wheat worldwide. William Som (WS), an Argentinian spring wheat landrace, has consistently exhibited high-level resistance to stripe rust for over 20 years in our field evaluations [...] Read more.

Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is a destructive disease of wheat worldwide. William Som (WS), an Argentinian spring wheat landrace, has consistently exhibited high-level resistance to stripe rust for over 20 years in our field evaluations in Washington state, USA. A previous study showed high-temperature adult-plant (HTAP) resistance in WS. To map the HTAP resistance quantitative trait loci (QTL) in WS, 114 F_5-8 recombinant inbred lines (RILs) from the cross AvS/WS were evaluated for their stripe rust response in seven field environments in Washington. The RILs and parents were genotyped with the Infinium 90K SNP chip. Four stable QTL, QYrWS.wgp-1BL on chromosome 1B (669–682 Mb), QyrWS.wgp-2AL on 2A (611–684 Mb), QyrWS.wgp-3AS on 3A (9–13 Mb), and QyrWS.wgp-3BL on 3B (476–535 Mb), were identified, and they explained 10.0–19.0%, 10.2–16.7%, 7.0–15.9%, and 12.0–27.8% of the phenotypic variation, respectively. The resistance in WS was found to be due to additive interactions of the four QTL. For each QTL, two Kompetitive allele-specific PCR (KASP) markers were developed, and these markers should facilitate the introgression of the HTAP resistance QTL into new wheat cultivars. Full article

(This article belongs to the Special Issue Advances in Plant Genomics and Genetics: 3rd Edition)

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15 pages, 3503 KiB

Open AccessArticle

Regulation of Vascular Calcification by M1-Type Macrophage-Derived Semaphorin 4D

by Hyun-Joo Park, Yeon Kim, Mi-Kyoung Kim, Hyung Joon Kim, Soo-Kyung Bae and Moon-Kyoung Bae

Int. J. Mol. Sci. 2025, 26(11), 5071; https://doi.org/10.3390/ijms26115071 (registering DOI) - 24 May 2025

Abstract

Vascular calcification is a critical pathological hallmark of cardiovascular diseases. Although previous studies have indicated that M1 macrophages significantly promote calcification, the exact underlying mechanisms remain unclear. This study examined whether semaphorin 4D (Sema4D), a class IV semaphorin involved in atherosclerosis development, is [...] Read more.

Vascular calcification is a critical pathological hallmark of cardiovascular diseases. Although previous studies have indicated that M1 macrophages significantly promote calcification, the exact underlying mechanisms remain unclear. This study examined whether semaphorin 4D (Sema4D), a class IV semaphorin involved in atherosclerosis development, is secreted by M1 macrophages and contributes to the calcification of vascular smooth muscle cells (VSMCs). We observed elevated expression and secretion of Sema4D in both M1 and M2 macrophages, with significantly higher levels in M1-polarized cells. M1 macrophages promoted VSMC calcification in both co-culture and conditioned medium systems, as evidenced by increased alkaline phosphatase activity, enhanced calcium deposition, and upregulation of osteogenic markers. Notably, neutralization of Sema4D in M1 conditioned medium using pepinemab, an anti-Sema4D antibody, effectively attenuated VSMC calcification induced by M1 macrophages. Conversely, supplementation of conditioned medium with recombinant Sema4D enhanced calcification and osteogenic signaling in VSMCs, further supporting the pro-calcifying role of Sema4D. Collectively, these findings highlight macrophage-derived Sema4D as a pivotal regulator of vascular calcification and a promising therapeutic target. Full article

(This article belongs to the Special Issue Smooth Muscle Cells in Vascular Disease)

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15 pages, 649 KiB

Open AccessArticle

Evaluating Glial Fibrillary Acidic Protein and Neurofilament Light as Potential Biomarkers for Spinocerebellar Ataxia 7

by Rana Hanna Al-Shaikh, Karen Jansen-West, Audrey Strongosky, Zoe Parrales, Judith A. Dunmore, Yuping Song, Tania F. Gendron, Juan C. Guevara, Helio A. G. Teive, Jarosław Dulski, Jarosław Sławek, Leonard Petrucelli, Zbigniew K. Wszolek and Mercedes Prudencio

Int. J. Mol. Sci. 2025, 26(11), 5070; https://doi.org/10.3390/ijms26115070 (registering DOI) - 24 May 2025

Abstract

Spinocerebellar ataxia type 7 (SCA7), a rare form of ataxia, possesses a wide phenotypic spectrum ranging from classic ataxic symptoms to blindness, multiorgan failure, cardiomyopathy, and early death among younger age groups. Biomarkers associated with disease progression and severity could aid in disease [...] Read more.

Spinocerebellar ataxia type 7 (SCA7), a rare form of ataxia, possesses a wide phenotypic spectrum ranging from classic ataxic symptoms to blindness, multiorgan failure, cardiomyopathy, and early death among younger age groups. Biomarkers associated with disease progression and severity could aid in disease prognostication. We evaluated the utility of glial fibrillary acidic protein (GFAP) and neurofilament light (NfL) in distinguishing patients with SCA7 from healthy controls and estimating patient prognosis. GFAP and NfL levels were measured in 23 plasma and 20 cerebrospinal fluid (CSF) samples from asymptomatic (N = 3) and symptomatic SCA7 participants (N = 10) and from healthy controls (N = 8). GFAP concentrations were elevated in the plasma (82.7 pg/mL) and CSF (9318 pg/mL) of patients with SCA7 compared to controls (plasma: 48.0 pg/mL; CSF: 89,056 pg/mL). Similarly, NfL plasma (21.6 pg/mL) and CSF (2615.0 pg/mL) levels were also significantly upregulated in SCA7 compared to controls (plasma: 8.2 pg/mL; CSF: 414.6 pg/mL). Higher levels of NfL, but not of GFAP, significantly discriminated symptomatic SCA7 patients from controls (area under de curve, AUC: 0.898, p = 0.0059, in plasma, and AUC: 1.0, p = 0.0012, in CSF). The levels of both biomarkers increased overtime, with plasma NfL levels strongly associated with a worse score in the scale for the assessment and rating of ataxia (SARA) (Spearman r: 0.8354, p = 0.0007; regression analysis: β: 0.021, 95% CI: 0.008–0.035, p = 0.0048). These findings suggest that NfL could serve as a valuable biomarker for monitoring disease progression and prognosis in SCA7 patients. Full article

(This article belongs to the Special Issue Advances in Rare Diseases Biomarkers: 2nd Edition)

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25 pages, 4271 KiB

Open AccessArticle

Cyclic GMP-AMP Synthase (cGAS) Deletion Promotes Less Prominent Inflammatory Macrophages and Sepsis Severity in Catheter-Induced Infection and LPS Injection Models

by Chatsuree Suksamai, Warerat Kaewduangduen, Pornpimol Phuengmaung, Kritsanawan Sae-Khow, Awirut Charoensappakit, Suwasin Udomkarnjananun, Sutada Lotinun, Patipark Kueanjinda and Asada Leelahavanichkul

Int. J. Mol. Sci. 2025, 26(11), 5069; https://doi.org/10.3390/ijms26115069 (registering DOI) - 24 May 2025

Abstract

Activation of cGAS, a cytosolic receptor recognizing double-stranded DNA, in macrophages is important in sepsis (a life-threatening condition caused by infection). The responses against sepsis induced by subcutaneous implantation of the Pseudomonas-contaminated catheters in cGAS-deficient (cGAS^−/−) mice were lower than [...] Read more.

Activation of cGAS, a cytosolic receptor recognizing double-stranded DNA, in macrophages is important in sepsis (a life-threatening condition caused by infection). The responses against sepsis induced by subcutaneous implantation of the Pseudomonas-contaminated catheters in cGAS-deficient (cGAS^−/−) mice were lower than in wild-type (WT) mice as indicated by liver enzymes, white blood cell count, cytokines, and M1-polarized macrophages in the spleens. Likewise, a lethal dose of lipopolysaccharide (LPS) induced less severe sepsis severity as determined by mortality, organ injury, cell-free DNA, and serum cytokines. Patterns of the transcriptome of lipopolysaccharide (LPS)-stimulated bone marrow-derived macrophages were clearly different between cGAS^−/− and WT cells. Gene set enrichment analysis (GSEA; a computational statistical determination of the gene set) indicated more prominent enrichment of oxidative phosphorylation (OXPHOS; the mitochondrial function) and mTORC1 pathways in LPS-activated cGAS^−/− macrophages compared with WT. Meanwhile, LPS upregulated cGAS and increased cGAMP (a cGAS inducer) only in WT macrophages along with less severe inflammation in cGAS^−/− macrophages, as indicated by supernatant cytokines, pro-inflammatory molecules (nuclear factor kappa B; NF-κB), M1 polarization (IL-1β, CD80, and CD86), and macrophage extracellular traps (METs; web-like structures composed of DNA, histones, and other proteins) through the detection of citrullinated histone 3 (CitH3) in supernatant and immunofluorescent visualization. In conclusion, less prominent pro-inflammatory responses of cGAS^−/− macrophages than WT were demonstrated in mice (catheter-induced sepsis and LPS injection model) and in vitro (transcriptomic analysis, macrophage polarization, and METs). Full article

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

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16 pages, 2991 KiB

Open AccessArticle

Anti-Tumor Activities of Anti-Siglec-15 Chimeric Heavy-Chain Antibodies

by Kexuan Cheng, Jiazheng Guo, Yating Li, Qinglin Kang, Rong Wang, Longlong Luo, Wei Wang and Jiansheng Lu

Int. J. Mol. Sci. 2025, 26(11), 5068; https://doi.org/10.3390/ijms26115068 (registering DOI) - 24 May 2025

Abstract

Immune checkpoint inhibitors like programmed cell death 1 (PD-1) antibodies have revolutionized cancer treatment, but patient response rates remain limited. Sialic acid-binding Ig-like lectin 15 (Siglec-15) has emerged as a promising new immune checkpoint target. Through phage display technology using a Bactrian camel [...] Read more.

Immune checkpoint inhibitors like programmed cell death 1 (PD-1) antibodies have revolutionized cancer treatment, but patient response rates remain limited. Sialic acid-binding Ig-like lectin 15 (Siglec-15) has emerged as a promising new immune checkpoint target. Through phage display technology using a Bactrian camel immunized with recombinant human Siglec-15, we generated six anti-Siglec-15 camelid nanobodies and constructed chimeric heavy-chain antibodies by fusing the VHH domains with human IgG-Fc. Following expression in HEK293-F cells and purification, three antibodies (S1, S5, S6) demonstrated specific binding to both human and murine Siglec-15 in ELISA and biolayer interferometry assays. In a xenograft model established by subcutaneous inoculation of NCI-H157-S15 cells into BALB/c nude mice, these antibodies showed distinct tumor targeting and significant blockade of Siglec-15 interactions with CD44, MAG, sialyl-Tn, and LRR4C ligands. All three antibodies exhibited anti-tumor effects, with S1 showing the most potent activity. S1-treated mice had significantly smaller tumor volumes and weights compared to controls. The S1, S5, and S6 treatment groups showed enhanced anti-tumor immunity, with reduced TGF-β, IL-6, and IL-10 levels. Notably, S1 treatment significantly increased tumor-associated macrophages in tumor tissues (p < 0.05). In conclusion, S1 exhibits remarkable anti-tumor activity and has the potential to be developed as a cancer immunotherapy targeting Siglec-15. Full article

(This article belongs to the Special Issue Immunomodulatory Molecules in Cancer)

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25 pages, 3919 KiB

Open AccessReview

Regulatory Mechanism of Intestinal Stem Cells Based on Hippo Pathway and Signaling Crosstalk in Chicken

by Tao Quan, Ran Li, Yaoxing Chen and Ting Gao

Int. J. Mol. Sci. 2025, 26(11), 5067; https://doi.org/10.3390/ijms26115067 (registering DOI) - 24 May 2025

Abstract

Recently, there has been a gradual increase in the demand for chicken and eggs. The gut, as the vital place of nutrient digestion and absorption, is highly associated with the development of livestock and poultry and the quality of meat, eggs, and milk. [...] Read more.

Recently, there has been a gradual increase in the demand for chicken and eggs. The gut, as the vital place of nutrient digestion and absorption, is highly associated with the development of livestock and poultry and the quality of meat, eggs, and milk. Intestinal stem cells, as an important source of intestinal cell proliferation and renewal, exert a vital effect on repairing injured intestinal epithelial cells and keeping homeostasis. Intestinal stem cell-regulated intestinal epithelial balance is closely controlled and modulated by interlinked developmental loops that maintain cell proliferation and differentiation processes in balance. Some conservative signaling pathways, including the Wnt, Notch, hedgehog, and bone morphogenetic protein (BMP) loops, have been proved to modulate intestinal health in poultry. Meanwhile, studies have revealed the importance of the Hippo pathway in gastrointestinal tract physiology by regulating intestinal stem cells. Moreover, crosstalk between Hippo and other signaling pathways provides tight, yet versatile, regulation of tissue homeostasis. In this review, we summarize studies on the role of the Hippo pathway in the intestine in these physiological processes and the underlying mechanisms responsible via interacting with these signaling pathways and discuss future research directions and potential therapeutic strategies targeting Hippo signaling in intestinal disease. A comprehensive understanding of how these signaling pathways regulate stem cell proliferation, differentiation, and self-renewal will help to understand the regulation of intestinal homeostasis. In addition, it has the capacity for creative ways to govern intestinal damage, enteritis, and associated disorders induced by different factors. Full article

(This article belongs to the Topic Recent Advances in Veterinary Pharmacology and Toxicology)

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36 pages, 16052 KiB

Open AccessArticle

The Role of Hydrogen Sulfide in the Localization and Structural–Functional Organization of p53 Following Traumatic Brain Injury: Development of a YOLO Model for Detection and Quantification of Apoptotic Nuclei

by Evgeniya Kirichenko, Stanislav Bachurin, Anton Lisovin, Rozaliia Nabiullina, Marya Kaplya, Aleksandr Romanov, Chizaram Nwosu and Stanislav Rodkin

Int. J. Mol. Sci. 2025, 26(11), 5066; https://doi.org/10.3390/ijms26115066 (registering DOI) - 24 May 2025

Abstract

Traumatic brain injury (TBI) triggers a cascade of molecular and cellular disturbances, including apoptosis, inflammation, and destabilization of neuronal connections. The transcription factor p53 plays a pivotal role in regulating cell fate following brain injury by initiating pro-apoptotic signaling cascades. Hydrogen sulfide (H [...] Read more.

Traumatic brain injury (TBI) triggers a cascade of molecular and cellular disturbances, including apoptosis, inflammation, and destabilization of neuronal connections. The transcription factor p53 plays a pivotal role in regulating cell fate following brain injury by initiating pro-apoptotic signaling cascades. Hydrogen sulfide (H₂S) may significantly contribute to the regulation of p53. Using scanning laser confocal microscopy, we found that after TBI, p53 accumulates extensively in the damaged cerebral cortex, showing distinct subcellular localization in neurons and astrocytes. In neurons, p53 predominantly localizes to the cytoplasm, suggesting involvement in mitochondria-dependent apoptosis, whereas in astrocytes, p53 is found in both the nucleus and cytoplasm, indicating possible activation of transcription-dependent apoptotic pathways. Quantitative analysis confirmed a correlation between p53 localization and morphological signs of cell death, as revealed by Sytox Green and Hoechst nuclear staining. Modulating H₂S levels exerted a marked influence on p53 expression and distribution. Administration of the H₂S donor sodium thiosulfate (Na₂S₂O₃) reduced the overall number of p53-positive cells, decreased nuclear localization, and lowered the level of apoptosis. Conversely, inhibition of H₂S synthesis using aminooxyacetic acid (AOAA) led to enhanced p53 expression, increased numbers of cells exhibiting nuclear fragmentation, and a more pronounced apoptotic response. These findings highlight a neuroprotective role for H₂S, likely mediated through the suppression of p53-dependent cell death pathways. To improve analytical accuracy, we developed a YOLO-based deep-learning model for the automated detection of fragmented nuclei. Additionally, evolutionary and molecular dynamics analysis revealed a high degree of p53 conservation among vertebrates and indicated that, although H₂S does not form stable complexes with p53, it may modulate its conformational dynamics. Full article

(This article belongs to the Section Biochemistry)

16 pages, 512 KiB

Open AccessReview

The Role of Helicobacter pylori Heat Shock Proteins in Gastric Diseases’ Pathogenesis

by Olga Maria Manna, Celeste Caruso Bavisotto, Melania Ionelia Gratie, Provvidenza Damiani, Giovanni Tomasello and Francesco Cappello

Int. J. Mol. Sci. 2025, 26(11), 5065; https://doi.org/10.3390/ijms26115065 (registering DOI) - 24 May 2025

Abstract

Helicobacter pylori (H. pylori) is a Gram-negative bacterium that colonizes the human stomach and is associated with several gastric diseases, including gastritis, peptic ulcer disease, and gastric cancer. The bacterium’s ability to thrive in the harsh gastric environment is due, to [...] Read more.

Helicobacter pylori (H. pylori) is a Gram-negative bacterium that colonizes the human stomach and is associated with several gastric diseases, including gastritis, peptic ulcer disease, and gastric cancer. The bacterium’s ability to thrive in the harsh gastric environment is due, to some extent, to its stress response mechanisms, with its heat shock proteins (HSPs) playing a putative, yet not fully understood, role in these adaptive processes. HSPs are a family of molecules, highly conserved throughout phylogenesis, that assist in protein folding, prevent aggregation, and ensure cellular homeostasis under stressful conditions. In H. pylori, HSPs contribute to survival in the stomach’s acidic environment and oxidative stress. Furthermore, they aid in the bacterium’s ability to adhere to gastric epithelial cells, modulate the host immune response, and form biofilms, all contributing to chronic infection and pathogenicity. The role of microbial HSPs in antibiotic resistance has also emerged as a critical area of research, as these proteins help stabilize efflux pumps, protect essential proteins targeted by antibiotics, and promote biofilm formation, thereby reducing the efficacy of antimicrobial treatments. Among bacterial HSPs, GroEL and DnaK are probably the major proteins that control most of the H. pylori’s functioning. Indeed, both proteins possess remarkable acid resistance, high substrate affinity, and dual roles in protein homeostasis and host interaction. These features make them critical for H. pylori’s adaptation, persistence, and pathogenicity in the gastric niche. In addition, recent findings have also highlighted the involvement of HSPs in the crosstalk between H. pylori and gastric epithelial cells mediated by the release of bacterial outer membrane vesicles and host-derived exosomes, both of these extracellular vesicles being part of the muco-microbiotic layer of the stomach and influencing cellular signalling and immune modulation. Considering their critical role in the survival and persistence of bacteria, microbial HSPs also represent potential therapeutic targets. Strategies aimed at inhibiting microbial HSP function, combined with conventional antibiotics or developing vaccines targeting microbial HSPs, could provide new avenues for the treatment of H. pylori infections and combat antibiotic resistance. This review explores the multifaceted roles of microbial HSPs in the pathogenesis of H. pylori, highlighting their contributions to bacterial adhesion, immune evasion, stress response, and antibiotic resistance. Full article

(This article belongs to the Special Issue Pathogenicity and Antibiotic Resistance of Helicobacter pylori)

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15 pages, 1438 KiB

Open AccessBrief Report

β-Hydroxybutyrate Reduces Body Weight by Modulating Fatty Acid Oxidation and Beiging in the Subcutaneous Adipose Tisue of DIO Mice

by Violeta Heras, Virginia Mela, Pallavi Kompella, Elena Rojano, Guillermo Paz-López, Lucia Hurtado-García, Almudena Ortega-Gómez, Maria José García-López, María Luisa García-Martín, Juan A. G. Ranea, Francisco J. Tinahones and Isabel Moreno-Indias

Int. J. Mol. Sci. 2025, 26(11), 5064; https://doi.org/10.3390/ijms26115064 (registering DOI) - 24 May 2025

Abstract

β-hydroxybutyrate (BHB) serves as an alternative cellular fuel during states of low glucose availability, such as fasting or carbohydrate restriction, when the body shifts to using fats and ketone bodies for energy. While BHB has shown potential metabolic benefits, its mechanisms of action [...] Read more.

β-hydroxybutyrate (BHB) serves as an alternative cellular fuel during states of low glucose availability, such as fasting or carbohydrate restriction, when the body shifts to using fats and ketone bodies for energy. While BHB has shown potential metabolic benefits, its mechanisms of action in the context of obesity are not fully understood. In this study, we examined the effects of BHB supplementation on subcutaneous adipose tissue (SAT) metabolism in a diet-induced obesity (DIO) mouse model. Adult male mice were first fed a high-fat diet for six weeks, followed by a standard diet with or without BHB supplementation for an additional six weeks. BHB supplementation led to significant body weight loss independent of food intake. This weight reduction was associated with decreased adipocyte differentiation, reflected by reduced peroxisome proliferator-activated receptor gamma (PPARγ) protein levels and lower uncoupling protein 1 (UCP1) expression, indicating altered SAT function. Transcriptomic analysis of SAT revealed upregulation of genes involved in fatty acid activation and transport (e.g., Slc27a2, Plin5, Acot4, Acsm3, Rik). Functional enrichment highlighted the activation of the PPAR signaling pathway and enrichment of peroxisomal components in the BHB group. Together, these results suggest that BHB promotes lipid remodeling in SAT, enhancing fatty acid metabolism while suppressing thermogenic pathways, and thus may represent a novel mechanism contributing to adiposity reduction and metabolic improvement. Full article

(This article belongs to the Topic Bioactive Compounds and Therapeutics: Molecular Aspects, Metabolic Profiles, and Omics Studies 2nd Edition)

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17 pages, 4963 KiB

Open AccessArticle

Characterization and Functional Analysis of a Novel Fungal Immunomodulatory Protein Gene from Ganoderma leucocontextum in B16-F10 Mouse Melanoma Cells

by Jiayi Yang, Mengyuan Jin, Lida Zhang, Yingying Wu and Xuanwei Zhou

Int. J. Mol. Sci. 2025, 26(11), 5063; https://doi.org/10.3390/ijms26115063 (registering DOI) - 24 May 2025

Abstract

Ganoderma leucocontextum, a newly identified species from the Tibetan Plateau, has been mainly studied for its polysaccharides and triterpenoids, with no prior reports on fungal immunomodulatory proteins (FIPs). This study explores the biological activity of FIP-gle2, cloned from G. leucocontextum and expressed [...] Read more.

Ganoderma leucocontextum, a newly identified species from the Tibetan Plateau, has been mainly studied for its polysaccharides and triterpenoids, with no prior reports on fungal immunomodulatory proteins (FIPs). This study explores the biological activity of FIP-gle2, cloned from G. leucocontextum and expressed in Pichia pastoris. The effects and mechanisms of recombinant FIP-gle2 (rFIP-gle2) on cell activity and melanin synthesis in mouse melanoma B16-F10 cells were investigated in vitro. The results showed that the FIP-gle2 gene, with an open reading frame (ORF) of 333 bp, encodes a 111-amino acid polypeptide with a molecular weight of 12.60 kDa and an isoelectric point of 4.48. We achieved a yield of 184.18 mg/L of rFIP-gle2. In vitro functional experiments showed that rFIP-gle2 significantly inhibited the proliferation of B16-F10 melanoma cells and induced apoptosis in a dose-dependent manner, particularly at concentrations above 1 μg/mL. At 3 μg/mL, rFIP-gle2 effectively inhibited tyrosinase activity and reduced melanin content, downregulating microphthalmia-associated transcription factor (MITF), tyrosinase (TYR), and tyrosinase-related proteins (TRP-1 and TRP-2). Furthermore, RNA-seq analysis indicated that differentially expressed genes in treated cells were enriched in the mitogen-activated protein kinase (MAPK) signaling pathway, with Western blotting confirming enhanced phosphorylation of JNK, ERK, and p38 proteins. Thus, P. pastoris is an effective host for rFIP-gle2 production, which shows potential for applications in pharmaceuticals, cosmeceuticals, and food fields. Full article

(This article belongs to the Special Issue Anticancer Drug Discovery Based on Natural Products)

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7 pages, 446 KiB

Open AccessArticle

Photophysical Properties of a Chiral Iridium-Based Photosensitizer as an Efficient Photodynamic Therapy Agent: A Theoretical Investigation

by Maciej Spiegel

Int. J. Mol. Sci. 2025, 26(11), 5062; https://doi.org/10.3390/ijms26115062 (registering DOI) - 24 May 2025

Abstract

This study employs time-dependent density functional theory to explore the photophysical properties of a chiral iridium(III) complex designed as a photosensitizer for photodynamic therapy. Key properties analyzed include one-photon absorption energies, singlet–triplet energy gaps, spin–orbit coupling constants, and intersystem crossing rate constants. The [...] Read more.

This study employs time-dependent density functional theory to explore the photophysical properties of a chiral iridium(III) complex designed as a photosensitizer for photodynamic therapy. Key properties analyzed include one-photon absorption energies, singlet–triplet energy gaps, spin–orbit coupling constants, and intersystem crossing rate constants. The potential for operation in a Type I PDT mechanism was assessed through ionization potential and electron affinity calculations. The results demonstrate that the complex is a promising PDT candidate, primarily operating in a Type II mechanism, while offering conditional viability for Type I photoreactivity under specific electronic and environmental conditions. Full article

(This article belongs to the Special Issue Exploring Organic Compounds in Photodynamic Therapy: Synthesis, Mechanism of Action and Biological Potential)

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13 pages, 1205 KiB

Open AccessArticle

Morphological Alterations of Conal Ridges and Differential Expression of AP2α in the Offspring Hearts of Experimental Diabetic Rats

by Tania Cristina Ramírez-Fuentes, Ricardo Jaime-Cruz, Carlos César Patiño-Morales, Laura Villavicencio-Guzmán, Juan Carlos Corona, María Cristina Revilla-Monsalve, Rosa Adriana Jarillo-Luna and Marcela Salazar-García

Int. J. Mol. Sci. 2025, 26(11), 5061; https://doi.org/10.3390/ijms26115061 (registering DOI) - 24 May 2025

Abstract

Neural crest cells (NCCs) play a significant role in the development of ventricular outflow tracts (OFTs), and cardiac neural crest cells (cNCCs) are involved in the development of the embryonic conus, suggesting that these cell lineages may be a teratogenic target for the [...] Read more.

Neural crest cells (NCCs) play a significant role in the development of ventricular outflow tracts (OFTs), and cardiac neural crest cells (cNCCs) are involved in the development of the embryonic conus, suggesting that these cell lineages may be a teratogenic target for the development of cardiopathies in offspring conceived under a hyperglycemic environment. We evaluate the effect of the hyperglycemic intrauterine environment on the morphological and anatomical changes in the conal ridges along with the alterations in the spatiotemporal expression of AP2α in offspring hearts at 13, 15, and 17 DPC. The anatomical and histological analysis of the hearts in the experimental group presented smaller dimensions compared to the control group in the offspring at the three ages studied. Consequently, this resulted in a hyperglycemic environment that altered the immunostaining of AP2α in the hearts of the offspring at the three ages studied. Thus, the hyperglycemic intrauterine environment in offspring caused important morphological alterations in the development of conal ridges and promoted the generation of conotruncal heart defects in which the double outlet of the right ventricle, the atrioventricular (AV) canal, predominated. Therefore, knowing that exposing the offspring to more glucose potentially can lead to complications during organogenesis of the circulatory and central nervous systems. Full article

(This article belongs to the Special Issue Molecular Advances in Gestational Diabetes Mellitus)

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34 pages, 6835 KiB

Open AccessReview

Endometrial Aging and Reproductive Decline: The Central Role of Mitochondrial Dysfunction

by Hiroshi Kobayashi, Miki Nishio, Mai Umetani, Hiroshi Shigetomi, Shogo Imanaka and Hiratsugu Hashimoto

Int. J. Mol. Sci. 2025, 26(11), 5060; https://doi.org/10.3390/ijms26115060 (registering DOI) - 24 May 2025

Abstract

Socioeconomic factors have led an increasing number of women to postpone childbirth, thereby elevating the risks of reduced fertility, pregnancy complications, preterm birth, cesarean delivery, and chromosomal abnormalities. While diminished oocyte quality is a well-established contributor to age-related infertility, endometrial dysfunction also plays [...] Read more.

Socioeconomic factors have led an increasing number of women to postpone childbirth, thereby elevating the risks of reduced fertility, pregnancy complications, preterm birth, cesarean delivery, and chromosomal abnormalities. While diminished oocyte quality is a well-established contributor to age-related infertility, endometrial dysfunction also plays a pivotal role. Optimizing both oocyte quality and endometrial health is essential for enhancing reproductive outcomes. Although aging has been defined by twelve hallmarks, research specifically addressing age-related changes in endometrial function remains limited. This review examines the process of endometrial aging, with a particular emphasis on mitochondrial function. A comprehensive literature search was conducted using PubMed and Google Scholar to identify relevant studies published up to 31 January 2025. Endometrial aging is driven by multiple biological mechanisms, most notably the decline in endometrial receptivity. Key contributing factors include hormonal dysregulation, chronic inflammation, cell cycle arrest, genomic instability, epigenetic alterations, telomere attrition, and mitochondrial dysfunction. Among these, mitochondrial dysfunction emerges as a central driver of the aging process. Endometrial senescence, precipitated by irreversible mitochondrial impairment, may underlie the progressive decline in reproductive potential. Elucidating the role of mitochondrial dysfunction in aging provides critical insights into the molecular basis of fertility decline, particularly through its impact on endometrial receptivity. Full article

(This article belongs to the Special Issue Mitochondrial Dysfunction: A Metabolic, Cardiovascular, Neurodegenerative and Neuromuscular Issue: 2nd Edition)

16 pages, 2344 KiB

Open AccessArticle

Circulating FGF21 and Ketone Bodies Modify the Risk of MASLD and Mortality: Insights from the PREVEND Cohort Study

by Mateo Chvatal-Medina, Yakun Li, Wendy A. Dam, Margery A. Connelly, Han Moshage, Stephan J. L. Bakker, Robin P. F. Dullaart and Adrian Post

Int. J. Mol. Sci. 2025, 26(11), 5059; https://doi.org/10.3390/ijms26115059 (registering DOI) - 24 May 2025

Abstract

Fibroblast growth factor 21 (FGF21) and ketone bodies are markers of metabolic dysregulation, independently associated with metabolic-dysfunction-associated steatotic liver disease (MASLD) and mortality. We studied their interaction with MASLD and all-cause mortality in 6025 participants from the Prevention of Renal and Vascular End-stage [...] Read more.

Fibroblast growth factor 21 (FGF21) and ketone bodies are markers of metabolic dysregulation, independently associated with metabolic-dysfunction-associated steatotic liver disease (MASLD) and mortality. We studied their interaction with MASLD and all-cause mortality in 6025 participants from the Prevention of Renal and Vascular End-stage Disease (PREVEND) cohort. Plasma FGF21 (immunoassay) and ketone body concentrations (nuclear magnetic resonance spectroscopy) were measured at baseline. A Fatty Liver Index ≥60 was used as a proxy of MASLD. Logistic regression assessed associations with MASLD, and Cox models evaluated all-cause mortality over a median follow-up of 10.3 years. FGF21 and ketone bodies were not correlated (r = 0.02, p = 0.06), but FGF21 (OR: 1.93 [1.81–2.05], p < 0.001) and ketone bodies (OR: 1.29 [1.19–2.05], p < 0.001) were independent of each other associated with MASLD, with a positive interaction (p = 0.004). Higher FGF21 (HR: 1.24, 95% CI: 1.16–1.32, p < 0.001) and ketone bodies (HR: 1.46, 95% CI: 1.34–1.59, p < 0.001) were associated with mortality, as well as with a positive interaction (p = 0.038). After adjustment for potential confounders, only ketone bodies remained independently associated, while the association of FGF21 became dependent on ketone body levels (interaction p = 0.005). These biomarkers may serve as integrated metabolic stress markers, improving risk stratification for MASLD and adverse outcomes. Full article

(This article belongs to the Special Issue Molecular Advances in Metabolic Dysfunction-Associated Steatotic Liver Disease)

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