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22 pages, 8108 KB  
Article
Melanin Deposition and Screening of Melanogenesis-Related Differential RNAs and Construction of ceRNA Regulatory Network in Liancheng White Ducks
by Wenli Shi, Li Li, Bangzhe Zhao, Qiannan Cai, Xiaopan Liu, Zhiming Zhu, Linli Zhang, Zhongwei Miao, Qinlou Huang, Nenzhu Zheng and Qingwu Xin
Animals 2026, 16(12), 1891; https://doi.org/10.3390/ani16121891 - 18 Jun 2026
Viewed by 247
Abstract
Liancheng white ducks have a distinctive “white feathers, black beak, and green feet” phenotype, making them a useful model for studying pigmentation traits in waterfowl. The previous study found that the F1 generation of Liancheng white ducks crossed with white-feathered ducks and hemp-feathered [...] Read more.
Liancheng white ducks have a distinctive “white feathers, black beak, and green feet” phenotype, making them a useful model for studying pigmentation traits in waterfowl. The previous study found that the F1 generation of Liancheng white ducks crossed with white-feathered ducks and hemp-feathered ducks were all gray-black in color. This indicates the specificity and complexity of melanin deposition in Liancheng white ducks, which makes the selection and breeding of pigment traits through phenotyping difficult. The aim of this study was to investigate the candidate transcriptomic regulatory signals of melanogenesis in Liancheng white ducks. Skin, mouth skin, foot skin, liver, and muscle samples were collected from 130-day-old Liancheng white ducks. Morphological differences were observed via histological analysis, and extraction-based pigment levels were determined. The results showed that melanin granules were clearly observed in tissues other than the liver and were distributed mainly in the basal layer of the epidermis and around feather follicles; the pigment values in the tissues decreased in the order mouth skin > liver > foot skin > muscle and skin. However, the relatively high liver value should be interpreted cautiously because obvious melanin granule deposition was not observed histologically. Whole-transcriptome sequencing was performed on mouth skin and skin samples. In total, 3074 differentially expressed genes (DEGs) were screened; upregulated genes associated with melanogenesis included melanocyte inducing transcription factor (MITF) and tyrosinase (TYR); downregulated genes included agouti signaling protein (ASIP) and adenylate cyclase 2 (ADCY2). Eighteen differentially expressed microRNAs (DEmiRNAs) were identified. Based on target prediction and pathway enrichment analysis, novel_290 and apl-miR-11588-3p were identified as candidate miRNAs potentially associated with melanogenesis-related pathways, and their predicted target genes included phosphatidylinositol 3-kinase (PI3K) and Janus kinase 1 (JAK1). Additionally, 364 differentially expressed long noncoding RNAs (DElncRNAs) were identified; TCONS_00063335 and TCONS_00019814 were identified as candidate lncRNAs potentially associated with melanogenesis-related genes, including TYR and TYRP1. A putative ceRNA network was constructed based on the predicted miRNA–mRNA and miRNA–lncRNA relationships, and ENSAPLT00000025522–apl-miR-11588-3p–MAPK8IP3 was identified as a candidate network relationship associated with MAPK-related pigmentation pathways. However, because this relationship was inferred mainly from bioinformatic prediction and expression association analysis, further functional validation is required to confirm whether it contributes to melanogenesis regulation. These findings provide candidate transcriptomic and noncoding RNA information for the further investigation of tissue-specific pigmentation in Liancheng white ducks. Full article
(This article belongs to the Section Animal Genetics and Genomics)
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11 pages, 343 KB  
Article
Maternal Genetic Variants and Gestational Duration: A Replication Study in a Japanese Cohort
by Rina Tanabu, Kaori Iino, Maki Sato, Mako Nakamura, Macthi Yokoyama, Yoshinori Tamada, Ken Itoh, Tatsuya Mikami, Koichi Murashita and Yoshihito Yokoyama
J. Clin. Med. 2026, 15(11), 4269; https://doi.org/10.3390/jcm15114269 - 1 Jun 2026
Viewed by 203
Abstract
Background/Objectives: The objective of this study is to evaluate whether maternal genetic variants previously associated with gestational duration in European-ancestry populations are associated with gestational duration in Japanese women. Methods: We analyzed 347 women with a history of delivery from a [...] Read more.
Background/Objectives: The objective of this study is to evaluate whether maternal genetic variants previously associated with gestational duration in European-ancestry populations are associated with gestational duration in Japanese women. Methods: We analyzed 347 women with a history of delivery from a community-based cohort in Aomori Prefecture, Japan. Gestational age at first delivery (weeks) was obtained from the Maternal and Child Health Handbook. Four representative maternal single-nucleotide polymorphisms were analyzed: rs2946169 near EBF1, rs2955117 in/near EEFSEC, rs12037376 in/near WNT4, and rs9861425 in/near ADCY5. Maternal genotype data were obtained from existing Japonica Array data generated as part of the Iwaki Health Promotion Project. Associations with gestational age at first delivery were tested using linear regression under an additive genetic model, adjusted for maternal age at first delivery. A Bonferroni-corrected threshold of p < 0.0125 was applied for four SNPs. Results: Mean gestational age at first delivery was 39.4 weeks. Each additional rs2946169 T allele was nominally associated with shorter gestation (adjusted β = −0.2679 weeks per allele; p = 0.021), but this association did not remain significant after Bonferroni correction. No significant associations were observed for rs2955117, rs12037376, or rs9861425. Conclusions: These findings provide suggestive evidence that maternal variation at the EBF1 locus may be related to gestational duration in Japanese women. However, the association did not remain significant after multiple-testing correction, and larger studies are needed to confirm this finding and clarify population-specific genetic effects. Full article
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24 pages, 3049 KB  
Article
From Transcriptional Reprogramming to Fat Quality Improvement: Dietary Artemisia ordosica Krasch. Optimizes Fatty Acid Profile in Cashmere Goats
by Lianguang Jiang, Yanli Zhao, Qingyue Zhang, Shangxiong Zhang, Xiaoyu Guo, Yongmei Guo and Sumei Yan
Animals 2026, 16(7), 1097; https://doi.org/10.3390/ani16071097 - 2 Apr 2026
Viewed by 1137
Abstract
This experiment investigated the effects of dietary Artemisia ordosica Krasch. (AOK) supplementation on the n3-polyunsaturated fatty acid (n3-PUFA) profile of subcutaneous adipose tissue (SADT) in Arbas cashmere goats and explored the underlying transcriptional mechanisms. Forty healthy, weaned kids (120 ± 10 days of [...] Read more.
This experiment investigated the effects of dietary Artemisia ordosica Krasch. (AOK) supplementation on the n3-polyunsaturated fatty acid (n3-PUFA) profile of subcutaneous adipose tissue (SADT) in Arbas cashmere goats and explored the underlying transcriptional mechanisms. Forty healthy, weaned kids (120 ± 10 days of age; similar body weight) were randomly allocated to two groups (n = 20): a control group (CON, basal diet) and an AOK group (AOK, basal diet with 3% of the roughage replaced by AOK). The feeding trial spanned 104 days, consisting of a 14-day adaptation period and 90 days of data acquisition. Compared with the CON group, AOK significantly reduced the content of saturated fatty acids (SFAs) and n6-polyunsaturated fatty acids (n6-PUFAs)/n3-PUFAs (n6/n3). In contrast, the levels of n3-PUFAs in the SADT of cashmere goats increased markedly (p < 0.05). Compared with the CON group, AOK exhibited significantly higher activities of hormone-sensitive lipase (HSL) (p = 0.027), adenylyl cyclase 2 (ADCY2) (p = 0.010), adenylyl cyclase 5 (ADCY5) (p = 0.046), cluster of differentiation 36 (CD36) (p = 0.013), solute carrier family 27 member 4 (SLC27A4) (p = 0.021), and fatty acid binding protein 4 (FABP4) (p = 0.040), along with significantly lower activities of fatty acid synthase (FAS) (p = 0.002), lipoprotein lipase (LPL) (p = 0.048), and stearoyl-coa desaturase (SCD) (p = 0.026) in SADT. Compared with the CON group, the activities of superoxide dismutase (SOD) (p = 0.032), catalase (CAT) (p = 0.010), glutathione peroxidase (GSH-PX) (p = 0.029), and total antioxidant capacity (T-AOC) (p = 0.002) were significantly increased in the AOK group. Transcriptomic profiling revealed that AOK supplementation downregulated mRNA levels of ADCY2, ADCY5, LPL, FAS, SCD, stearoyl-CoA desaturase 1 (SCD1), stearoyl-CoA desaturase 2 (SCD2), glycogen synthase 1 (GYS1), acyl-CoA oxidase 1 (ACOX1), acetyl-CoA carboxylase (ACC), diacylglycerol acyltransferase 1 (DGAT1), fatty acid desaturase 1 (FADS1), solute carrier family 27 member 2 (SLC27A2), erythroblastic leukemia viral oncogene homolog 4 (ERBB4), and carnitine palmitoyltransferase 1B (CPT1B) (p < 0.05). It also markedly induced acyl-CoA synthetase long-chain family member 4 (ACSL4) (p < 0.01) in SADT. Genes significantly enriched in the adenosine-monophosphate-activated protein kinase (AMPK) signaling pathway included LPL, SCD1, CPT1B, and GYS1 (p = 0.010). Genes significantly enriched in the phosphatidylinositol 3-kinase-akt (PI3K-Akt) signaling pathway included GYS1 and ERBB4 (p = 0.015). CPT1B, ADCY2, and GYS1 were identified as the genes significantly enriched in the insulin resistance signaling pathway (p = 0.048). LPL was the only gene significantly enriched in the cholesterol metabolism pathway (p = 0.049). Genes showing a tendency toward significant enrichment in the peroxisome-proliferator-activated receptor (PPAR) signaling pathway included ACSL4, CPT1B, SCD1, and LPL (p = 0.051). These interconnected cascades improve insulin sensitivity, stimulate triglyceride (TG) hydrolysis, and modulate n3-PUFA levels. Supplementation with AOK enhances n3-PUFA content by accelerating TG breakdown while simultaneously restraining FA oxidation in SADT. Consequently, AOK supplementation can be effectively used to enhance the nutritional value of cashmere goat meat through improved n3-PUFA deposition in SADT. Full article
(This article belongs to the Section Small Ruminants)
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19 pages, 4796 KB  
Article
Enhanced Toxicity Induced by Combined Exposure to Neonicotinoid Insecticides and Fluoroquinolone Antibiotics in Human Neuroblastoma SK-N-SH Cells
by Gulijiazi Yeerkenbieke, Tao Wang, Yun Yang, Shuai Shi and Xiaoxia Lu
Toxics 2026, 14(3), 195; https://doi.org/10.3390/toxics14030195 - 25 Feb 2026
Viewed by 999
Abstract
Neonicotinoid insecticides and fluoroquinolone antibiotics frequently co-occur in aquatic and terrestrial environments, posing a threat to human health, yet their combined neurotoxic potential remains poorly characterized. This study aimed to assess the cytotoxicity of typical neonicotinoids and fluoroquinolones as well as their mixtures [...] Read more.
Neonicotinoid insecticides and fluoroquinolone antibiotics frequently co-occur in aquatic and terrestrial environments, posing a threat to human health, yet their combined neurotoxic potential remains poorly characterized. This study aimed to assess the cytotoxicity of typical neonicotinoids and fluoroquinolones as well as their mixtures in human neuroblastoma SK-N-SH cells and identify affected pathways. SK-N-SH cells were exposed to clothianidin (CLO), imidacloprid (IMI), enrofloxacin (ENR), and ofloxacin (OFX) individually and in fixed-ratio mixtures (50% of each compound’s IC50) for 24 h and 48 h, and cell viability was quantified using the alamarBlue® method. Single-compound dose–response testing showed time-dependent cytotoxicity, with higher potency for fluoroquinolones (24 h IC50: ENR 1.446 mM, OFX 2.742 mM; 48 h IC50: ENR 0.826 mM, OFX 2.005 mM) than neonicotinoids (24 h IC50: IMI 4.754 mM, CLO 5.356 mM; 48 h IC50: IMI 3.631 mM, CLO 4.029 mM). Concentration-addition analysis indicated that most mixtures produced synergistic interaction in reduction in cell viability, with ENR+OFX showing the strongest effect at 48 h (Observed viability 7.138% vs. Predicated viability 82.368%). RNA-seq (24 h) revealed that binary mixtures generally induced more differentially expressed genes than single exposures, and ENR-containing mixtures showed the largest transcriptomic shifts, enriching pathways related to cellular stress and injury as well as neuronal signaling and connectivity. RT-qPCR validated the changes in expressions of five key neurobiology-relevant genes (LMO3, NOS1, ADCY8, FGF7 and TNFRSF12A). These findings highlight the importance of assessing insecticide–antibiotic mixtures when evaluating their hazards in environment. Full article
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20 pages, 3960 KB  
Article
Transcriptome Analysis of Different Stages in the Early Ovarian Development of the Greater Amberjack (Seriola dumerili)
by Qiuxia Deng, Yang Huang, Xiaoying Ru, Haoyi Lin, Xinxin You, Ruijuan Hao, Hang Li, Qin Hu, Dongying Zhang, Yanfei Zhao, Jinhui Wu, Guangli Li and Chunhua Zhu
Animals 2026, 16(5), 709; https://doi.org/10.3390/ani16050709 - 25 Feb 2026
Viewed by 682
Abstract
The greater amberjack (Seriola dumerili) is a globally important aquaculture fish species belonging to the family Carangidae. However, current research on the ovarian development and regulation of the greater amberjack is limited, particularly with regard to early ovarian development in artificially [...] Read more.
The greater amberjack (Seriola dumerili) is a globally important aquaculture fish species belonging to the family Carangidae. However, current research on the ovarian development and regulation of the greater amberjack is limited, particularly with regard to early ovarian development in artificially bred individuals. Therefore, this study aims to analyze the ovarian transcriptomes at the primary growth stage in one-year-old fish at the chromatin nucleolar stage and two-year-old fish at the perinucleolar stage to suggest how early ovarian development takes place in artificially bred greater amberjacks. To do this, the study constructed ovarian mRNA expression profiles of different stages of artificially bred fish and identified differentially expressed genes (DEGs), Gene Ontology terms, and Kyoto Encyclopedia of Genes and Genomes pathways important for various physiological processes. A functional analysis revealed that the DEGs closely related to ovarian development were involved in cell growth and death (e.g., ccnd1, cdk1, cdc6, cdc25b, fbxo43, cdkn1a and cdkn1d), the TGF-beta signaling pathway (e.g., gdf6, bmpr1b, gsdf, amh, tgfb3, and smad6), steroid hormone biosynthesis (e.g., hsd3b1, hsd11b2, hsd17b12 and hsd20b2), and the endocrine system (e.g., mapk8a, mapk11, mapk12a, mapk13 and adcy9). These findings suggest that ovarian development is the result of the coordinated regulation of various genes responsible for various functions. This study provides a theoretical basis for exploring the underlying molecular mechanisms of early ovarian development in artificially bred greater amberjacks. Full article
(This article belongs to the Section Aquatic Animals)
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14 pages, 2810 KB  
Article
ADCY9 Regulates Neural Stem Cells Via Mitofusin-1 to Maintain Planarian (Dugesia japonica) Cephalic Ganglia Regeneration
by Xinrui Wang, Sitong Hu, Ruijia Zhang, Xinlu Han, Lili Gao, Fengtang Yang, Zhonghong Cao and Hui Zhen
Cells 2026, 15(5), 389; https://doi.org/10.3390/cells15050389 - 24 Feb 2026
Viewed by 740
Abstract
ADCY9, a crucial member of the adenylate cyclase family, exerts neuroprotective and analgesic effects in the nervous system by modulating the activity of the cAMP/AMPK signaling pathway. However, the role of the ADCY9 gene in neural regeneration remains unreported. In this study, [...] Read more.
ADCY9, a crucial member of the adenylate cyclase family, exerts neuroprotective and analgesic effects in the nervous system by modulating the activity of the cAMP/AMPK signaling pathway. However, the role of the ADCY9 gene in neural regeneration remains unreported. In this study, we utilized Dugesia japonica, a highly regenerative planarian species, as a model to systematically examine the spatiotemporal expression pattern of the ADCY9 gene during planarian brain regeneration and investigate its regulatory function in this process. The results demonstrated that the downregulation of ADCY9 resulted in abnormal brain regeneration in planarians, characterized by partial loss of the nerve cord, reduced numbers of collateral branches, and significant inhibition of the regeneration and differentiation of multiple neuron types. RNA sequencing revealed that the downregulation of ADCY9 led to 499 differentially expressed genes, with KEGG enrichment pathway analysis indicating significant associations with neurodegenerative diseases such as Alzheimer’s disease and Parkinson’s disease. Double RNAi experiments demonstrated that simultaneous knockdown of ADCY9 and Mitofusin-1 significantly restored neural regeneration. Collectively, ADCY9 might promote the comprehensive reconstruction of neural structure by hierarchically regulating the regeneration intensity through negative regulation of the downstream inhibitory factor Mitofusin-1. This study discloses the function of ADCY9 in planarian neural regeneration, providing a theoretical foundation for its application in investigating neural regeneration mechanisms and neurodegenerative disease pathogenesis in higher vertebrates. Full article
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24 pages, 6258 KB  
Article
Psoralen Promotes Direct Chemical Reprogramming of Mouse Embryonic Fibroblasts into Osteoblast-like Cells
by Wenjie Li, Haixia Liu, Xinyu Wan, Ding Cheng, Ruyuan Zhu and Zhiguo Zhang
Pharmaceutics 2026, 18(2), 279; https://doi.org/10.3390/pharmaceutics18020279 - 23 Feb 2026
Cited by 1 | Viewed by 939
Abstract
Background/Objectives: Cells derived from direct chemical reprogramming into osteoblasts represent a promising source for bone regeneration, but the efficiency needs improvement. Here, we systematically evaluated whether the natural compound psoralen (Psr) could enhance this process and explored its therapeutic potential and mechanism [...] Read more.
Background/Objectives: Cells derived from direct chemical reprogramming into osteoblasts represent a promising source for bone regeneration, but the efficiency needs improvement. Here, we systematically evaluated whether the natural compound psoralen (Psr) could enhance this process and explored its therapeutic potential and mechanism of action. Methods: Mouse embryonic fibroblasts (MEFs) were treated with a cocktail of forskolin and phenamil (FP), supplemented with Psr. In vitro differentiation was assessed by alkaline phosphatase and Alizarin Red S staining, reverse transcription quantitative PCR, immunofluorescence and Western blot. The bone-regenerative potential of the derived chemically induced osteoblast-like cells (ciOBs) was evaluated in critical-sized calvarial defects, femoral cortical defects and a subcutaneous ectopic implantation model, using micro-computed tomography and histology. Mechanistic insights of Psr were gained by analyzing the adenylyl cyclase 9 (ADCY9)/cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA)/cAMP response element-binding protein (CREB) axis using inhibitor SQ22536. Results: Psr acted synergistically with the FP cocktail to drive efficient osteogenic reprogramming of MEFs. At an optimal concentration of 25 μM, Psr enabled the most robust induction of early osteogenic markers and generation of mature, mineralizing ciOBs in vitro. In vivo, FP + Psr-induced ciOBs repaired critical-sized calvarial and femoral cortical defects and generated substantial, vascularized bone tissue in ectopic sites. Mechanistically, Psr co-treatment potently activated the ADCY9/cAMP/PKA/CREB pathway, and pharmacological inhibition of this pathway completely abolished the pro-osteogenic effects of Psr. Conclusions: Psr acts as a potent synergistic enhancer of direct chemical reprogramming, generating functional osteoblast-like cells with robust bone-regenerative capacity via activation of the ADCY9/cAMP/PKA/CREB pathway. Full article
(This article belongs to the Section Biopharmaceutics)
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18 pages, 4326 KB  
Article
DCS: A Zero-Shot Anomaly Detection Framework with DINO-CLIP-SAM Integration
by Yan Wan, Yingqi Lang and Li Yao
Appl. Sci. 2026, 16(4), 1836; https://doi.org/10.3390/app16041836 - 12 Feb 2026
Viewed by 1313
Abstract
Recently, the progress of foundation models such as CLIP and SAM has shown the great potential of zero-shot anomaly detection tasks. However, existing methods usually rely on general descriptions such as “abnormal”, and the semantic coverage is insufficient, making it difficult to express [...] Read more.
Recently, the progress of foundation models such as CLIP and SAM has shown the great potential of zero-shot anomaly detection tasks. However, existing methods usually rely on general descriptions such as “abnormal”, and the semantic coverage is insufficient, making it difficult to express fine-grained anomaly semantics. In addition, CLIP primarily performs global-level alignment, and it is difficult to accurately locate minor defects, while the segmentation quality of SAM is highly dependent on prompt constraints. In order to solve these problems, we proposed DCS, a unified framework that integrates Grounding DINO, CLIP and SAM through three key innovations. First of all, we introduced FinePrompt for adaptive learning, which significantly enhanced the modeling ability of exception semantics by building a fine-grained exception description library and adopting learnable text embeddings. Secondly, we have designed an Adaptive Dual-path Cross-modal Interaction (ADCI) module to achieve more effective cross-modal information exchange through dual-path fusion. Finally, we proposed a Box-Point Prompt Combiner (BPPC), which combines box prior information provided by DINO with the point prompt generated by CLIP, so as to guide SAM to generate finer and more complete segmentation results. A large number of experiments have proved the effectiveness of our method. On the MVTec-AD and VisA datasets, DCS has achieved the most state-of-the-art zero-shot anomaly detection results. Full article
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24 pages, 13198 KB  
Article
Multi-Omics Profiling of the Hepatopancreas of Ridgetail White Prawn Exopalaemon carinicauda Under Sulfate Stress
by Ruixuan Wang, Chen Gu, Hui Li, Libao Wang, Ruijian Sun, Kuipeng Fu, Wenjun Shi and Xihe Wan
Int. J. Mol. Sci. 2026, 27(2), 1056; https://doi.org/10.3390/ijms27021056 - 21 Jan 2026
Viewed by 706
Abstract
With intensifying global climate change and human activities, and with regional topography interactions, soil and water salinization has intensified, posing major ecological and environmental challenges worldwide. Here, we integrated histology, transmission electron microscopy, RNA sequencing (RNA-seq) and data-independent acquisition (DIA)-based proteomics to profile [...] Read more.
With intensifying global climate change and human activities, and with regional topography interactions, soil and water salinization has intensified, posing major ecological and environmental challenges worldwide. Here, we integrated histology, transmission electron microscopy, RNA sequencing (RNA-seq) and data-independent acquisition (DIA)-based proteomics to profile hepatopancreas responses of Exopalaemon carinicauda during acute sulfate stress (≤48 h). Sulfate exposure disrupted tubular architecture and organelle integrity, consistent with early cellular injury. Multi-omics analyses revealed metabolic reprogramming marked by suppressed glycolysis (e.g., HK2, ENO) and enhanced oxidative phosphorylation (e.g., ATP5F1B), together with activation of calcium signaling (e.g., SLC8A1, ADCY9) and reinforcement of antioxidant/one-carbon and glucose-branch pathways (e.g., SHMT2, PGAM2). These coordinated transcript–protein changes indicate a shift from rapid cytosolic ATP supply to mitochondrial ATP production while buffering Ca2+ overload and reactive oxygen species. Collectively, our results delineate the physiological and molecular adjustments that enable E. carinicauda to cope with sulfate conditions and provide mechanistic targets for selective breeding and water-quality management in saline–alkaline aquaculture. Full article
(This article belongs to the Section Molecular Genetics and Genomics)
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26 pages, 3486 KB  
Article
Evaluation of Anticancer Potential of Ganoderma lucidum on MCF-7 Breast Cancer Cells Through Genetic Transcription of Energy Metabolism
by Levent Gülüm, Emrah Güler, Emir Çapkınoğlu, Ayşe Büşranur Çelik and Yusuf Tutar
Antioxidants 2025, 14(12), 1471; https://doi.org/10.3390/antiox14121471 - 8 Dec 2025
Cited by 5 | Viewed by 2266
Abstract
Ganoderma lucidum has long been recognized for its medicinal properties, particularly due to its antioxidant, anti-inflammatory, and pro-apoptotic components such as polysaccharides and triterpenoids. This study aimed to evaluate the cytotoxic and molecular effects of ethanol and methanol extracts of G. lucidum as [...] Read more.
Ganoderma lucidum has long been recognized for its medicinal properties, particularly due to its antioxidant, anti-inflammatory, and pro-apoptotic components such as polysaccharides and triterpenoids. This study aimed to evaluate the cytotoxic and molecular effects of ethanol and methanol extracts of G. lucidum as well as doxorubicin on MCF-7 breast cancer cells. The cytotoxicity was assessed via MTT assay. The methanol extract showed stronger cytotoxicity (IC50: 62.37 µg/mL) than the ethanol extract, while doxorubicin exhibited an IC50 value of 0.66 mM. Phenolic profiling by HPLC revealed high levels of vanillic acid, gallic acid and (−)-epicatechin in the methanol extract, while volatile compounds such as hexanal and acetic acid were identified by GC-MS. Flow cytometric analysis demonstrated G0/G1 phase cell cycle arrest and an increase in early and late apoptotic populations. Gene expression studies using RT-qPCR showed significant downregulation of ACAT1, ADCY3, and NME2, key regulators of energy metabolism and epigenetic modification. On the other hand, doxorubicin treatment upregulated ACAT1 and ADCY3, while a slight downregulation was observed in NME2. These molecular changes suggest that G. lucidum induces apoptosis and impairs cancer cell proliferation through metabolic disruption and gene modulation. Full article
(This article belongs to the Section Health Outcomes of Antioxidants and Oxidative Stress)
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16 pages, 2801 KB  
Article
Enterococcus faecium WEFA23-Derived Surface Layer Protein OTC Prevents Listeria monocytogenes Invasion by Strengthening Intestinal Barrier Function and Modulating Immune Responses
by Yao He, Bing Dong, Ke Xie, Yingsheng Hu, Yina Huang, Xueying Tao and Hua Wei
Foods 2025, 14(23), 4110; https://doi.org/10.3390/foods14234110 - 30 Nov 2025
Cited by 1 | Viewed by 676
Abstract
Listeria monocytogenes (L. monocytogenes) is a major foodborne pathogen which can invade intestinal epithelial cells and cause severe systemic infection. Probiotics, as well as their surface layer proteins, hold broad promise for enhancing intestinal barrier function and defending against pathogenic invasion. [...] Read more.
Listeria monocytogenes (L. monocytogenes) is a major foodborne pathogen which can invade intestinal epithelial cells and cause severe systemic infection. Probiotics, as well as their surface layer proteins, hold broad promise for enhancing intestinal barrier function and defending against pathogenic invasion. In the present study, the antagonistic effects of surface layer protein ornithine carbamoyltransferase (OTC) from Enterococcus faecium (E. faecium) WEFA23 against L. monocytogenes were systematically evaluated in vitro in human intestinal epithelial Caco-2 cells, including assessments of anti-adhesion and anti-invasion capacity, inflammatory cytokine responses, intestinal barrier integrity, and transcriptomic changes, by comparing the effects of wild-type E. faecium WEFA23 and a previously constructed E. faecium WEFA23 otc gene knockout strain (E. faecium WEFA23 otc−/−). The results demonstrated that E. faecium WEFA23 achieved significant stronger anti-adhesion and anti-invasion capacity of L. monocytogenes (p < 0.05) in the presence of OTC, potentially through increasing tight junction protein expression, regulating inflammatory cytokines, and modulating the virulence factors of the pathogen. To elucidate the potential mechanism of the inhibitory effect of OTC protein, RNA-seq was performed. The results revealed that the significantly regulated core differentially expressed genes (DEGs), including ADCY2, OARI3, CCL5, and CXCL9, were found to be involved in γ-aminobutyric acid (GABA)-ergic synapse, calcium, and toll-like receptor signaling pathways. These findings demonstrated that OTC is involved in blocking Listeria invasion and revealed the function of the OTC from E. faecium WEFA23 in antimicrobial and intestinal mucosal defense, providing a conceptual foundation for the development of new probiotic intervention strategies in anti-infection. Full article
(This article belongs to the Special Issue Lactic Acid Bacteria: The Functions and Applications in Foods)
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18 pages, 4537 KB  
Article
Study of FOXL2 Regulation on Ovarian Function in Chlamys farreri Through Comparative ChIP-Seq and Transcriptome Analysis Using RNA Interference
by Xiaoling Liu, Han Yun, Yan Xing, Shuo Wang, Xueying Zhou and Jianbai Zhang
Biology 2025, 14(9), 1259; https://doi.org/10.3390/biology14091259 - 12 Sep 2025
Viewed by 1387
Abstract
FOXL2 (forkhead box protein L2) is a transcription factor, its function and regulatory mechanism have been mainly studied in mammals; related research on marine invertebrates is still insufficient. It was found that oogenesis was affected, and even a small number of cells resembling [...] Read more.
FOXL2 (forkhead box protein L2) is a transcription factor, its function and regulatory mechanism have been mainly studied in mammals; related research on marine invertebrates is still insufficient. It was found that oogenesis was affected, and even a small number of cells resembling spermatogonial morphology appeared in C. farreri ovaries after the FOXL2 was knocked down through RNA interference (RNAi) technology in our laboratory previously. Based on previous research, this paper conducted transcriptome sequencing and differential expression analysis on the ovarian tissues between the experimental group (post-RNAi) and the control group (pre-RNAi) of C. farreri, and used recombinant C. farreri FOXL2 protein for antibody production in Chromatin Immunoprecipitation Sequencing (ChIP seq) experiments to comprehensively analyze the pathways and key genes regulated by FOXL2 during oogenesis. The results showed that in the RNAi experimental group, 389 genes were upregulated, and 1615 genes were downregulated. Among the differentially expressed genes (DEGs), the differential genes related to gender or gonadal development are relatively concentrated in physiological processes such as steroid hormone synthesis, spermatogenesis, gonadal development, and ovarian function maintenance, as well as the FoxO and estrogen signaling pathways. Combining transcriptome and ChIP-seq data, it was found that there were some genes related to sex gonadal development among genes which were directly regulated by FOXL2, such as Wnt4, SIRT1, HSD17B8, GABABR1, KRAS, NOTCH1, HSD11B1, cPLA2, ADCY9, IP3R1, PLCB4, and Wnt1. This study lays the foundation for a deeper understanding of the FOXL2′s specific regulatory mechanism during oogenesis in scallops as a transcription factor. Full article
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23 pages, 8967 KB  
Article
Comprehensive Analysis of N6-Methyladenosine Methylation in Transverse Aortic Constriction-Induced Cardiac Fibrosis Based on MeRIP-Seq Analysis
by Shidong Liu, Pengying Zhao, Yuyuan He, Jieneng Wang, Bing Song and Cuntao Yu
Biomedicines 2025, 13(9), 2092; https://doi.org/10.3390/biomedicines13092092 - 27 Aug 2025
Cited by 2 | Viewed by 1804
Abstract
Background: The function and mechanism of N6-methyladenosine (m6A) methylation in pressure-overload cardiac fibrosis remains limited and unclear. This study aims to analyze and predict m6A modifications present in mouse hearts because of transverse aortic constriction (TAC). Materials and Methods: Twelve male C57BL/6 mice [...] Read more.
Background: The function and mechanism of N6-methyladenosine (m6A) methylation in pressure-overload cardiac fibrosis remains limited and unclear. This study aims to analyze and predict m6A modifications present in mouse hearts because of transverse aortic constriction (TAC). Materials and Methods: Twelve male C57BL/6 mice were randomly assigned to two groups, TAC group and sham group. The RNA Dot Blot assay was employed to evaluate the overall m6A methylation levels in both TAC and sham mice. The expression level of m6A-related enzymes were investigated through RT-PCR and Western blotting. MeRIP-seq and RNA-seq analyses were conducted to identify differentially modified m6A genes and mRNA expression genes. The protein–protein interaction (PPI) network was carried out to choose potential hub genes. Additionally, the transcription factor (TF)–microRNA (miRNA) coregulatory network and the drug–hub gene interaction network were built based on these hub genes. Furthermore, molecular docking simulations were also performed to analyze the interactions between drugs and hub genes. Results: Compared with the sham group, the TAC group demonstrated elevated levels of global m6A methylation. METTL3 and METTL14 were significantly upregulated, whereas FTO and ALKBH5 were significantly downregulated following TAC. MeRIP-seq analysis identified 17,806 m6A peaks associated with 9184 genes and 16,392 m6A peaks associated with 8550 genes in the TAC and sham groups, respectively. In conjunction with RNA-seq data, 66 genes were identified as exhibiting concurrent differences in both m6A methylation levels and mRNA expression. Six hub genes, Cd33, Irf4, Nr4a2, Hspa1b, Nr4a1, and Adcy1, were identified through the construction of a PPI network. The TF-miRNA coregulatory network contains six hub genes, 31 miRNAs, and 24 TFs. The drug–hub genes interaction network included five hub genes and 36 candidate drugs. Conclusions: The m6A modification is prevalent in TAC-induced cardiac fibrosis and significantly contributes to the fibrotic process by regulating critical genes. In the future, it may emerge as one of the potential cardiac fibrosis therapeutic targets. Full article
(This article belongs to the Section Cell Biology and Pathology)
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19 pages, 1520 KB  
Article
Impact of Vitamin B12 Supplementation on Cardiovascular Health in the Silver Star Bamboo Rat, a Species That Feeds Primarily on Bamboo
by Lei Chen, Zhoulong Chen, Yongqi Zhao, Nan Yang, Jingheng Wang, Yanni Zhao, Lijun Luo and Xiuyue Zhang
Animals 2025, 15(17), 2526; https://doi.org/10.3390/ani15172526 - 27 Aug 2025
Viewed by 1613
Abstract
Specialized herbivores like giant pandas (Ailuropoda melanoleuca), red pandas (Ailurus fulgens), and bamboo rats, which primarily consume bamboo, are at risk of nutrient deficiencies, particularly vitamin B12 (VB12), potentially leading to cardiovascular diseases. This study explored the effects of [...] Read more.
Specialized herbivores like giant pandas (Ailuropoda melanoleuca), red pandas (Ailurus fulgens), and bamboo rats, which primarily consume bamboo, are at risk of nutrient deficiencies, particularly vitamin B12 (VB12), potentially leading to cardiovascular diseases. This study explored the effects of VB12 supplementation on cardiovascular health in silver star bamboo rats (Rhizomys pruinosus). We first conducted a comprehensive genome annotation of R. pruinosus, laying the foundation for in-depth evolutionary studies. Comparative transcriptomic analysis revealed that genes related to cardiovascular disease (e.g., Sgcb, Adcy2, Itga1, Itgb8, Ifng, and Gpc1) were upregulated in the livers of R. pruinosus compared to carnivorous and omnivorous rodents, indicating a higher cardiovascular disease risk. After 60 days of VB12 supplementation, liver transcriptome analysis revealed significant improvements in cardiovascular health markers, including reduced cholesterol synthesis and enhanced fatty acid metabolism. Serum biochemical assays indicated that VB12 supplementation led to reduced homocysteine levels, decreased low-density lipoprotein (LDL)-to-high-density lipoprotein (HDL) ratios, and increased the apolipoprotein A-to-apolipoprotein B ratio. These findings suggest that VB12 may mitigate cardiovascular disease risk and could be considered in the dietary management of specialized bamboo-eating species. Our study provides valuable insights into disease prevention strategies for these species with similar dietary habits. Full article
(This article belongs to the Special Issue Feed Additives in Animal Nutrition)
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35 pages, 19403 KB  
Article
Effects of Temperature and Salinity on Ovarian Development and Differences in Energy Metabolism Between Reproduction and Growth During Ovarian Development in the Lateolabrax maculatus
by Yangtao Peng, Lulu Yan, Chao Zhao, Bo Zhang, Bo Zhang and Lihua Qiu
Int. J. Mol. Sci. 2025, 26(17), 8295; https://doi.org/10.3390/ijms26178295 - 27 Aug 2025
Cited by 3 | Viewed by 2362
Abstract
Fish reproduction requires suitable salinity and temperature, as well as sufficient energy. This study investigated temperature and salinity effects on ovarian development of Lateolabrax maculatus and energy metabolism differences between reproduction and growth. Two salinities (4‰ and 30‰) and temperatures (18 ± 1 [...] Read more.
Fish reproduction requires suitable salinity and temperature, as well as sufficient energy. This study investigated temperature and salinity effects on ovarian development of Lateolabrax maculatus and energy metabolism differences between reproduction and growth. Two salinities (4‰ and 30‰) and temperatures (18 ± 1 °C and 30 ± 1 °C) formed four treatments: SWNT (30‰, 30 ± 1 °C), SWLT (30‰, 18 ± 1 °C), FWLT (4‰, 18 ± 1 °C), and FWNT (4‰, 30 ± 1 °C). GSI and sex hormones (FSH, LH, E2, and 17α,20β-DHP) were measured. Transcriptome analysis explored how temperature and salinity regulate ovarian development in L. maculatus, while integrated transcriptomic and targeted energy metabolomic analyses revealed energy metabolism differences between ovary and muscle during this process. The results showed that low salinity (4‰) and low temperature (18 ± 1 °C) synergistically promoted ovarian development in the FWLT group, as indicated by a significant increase in GSI and elevated levels of key sex hormones (FSH, LH, E2, and 17α,20β-DHP). Transcriptome analysis showed that low temperature activated pathways involved in steroidogenesis, oocyte maturation, and meiosis, and genes such as ADCY6, PRKACB, CPEB4, FZD7-A, and CCND2 were significantly upregulated. Salinity changes mainly affected amino acid metabolism, cholesterol metabolism, and the insulin signaling pathway. Genes such as PCSK9 and CKM may regulate ovarian development by regulating hormone synthesis and energy metabolism. Comprehensive transcriptome and metabolome analyses show that glycolysis is downregulated and oxidative phosphorylation is upregulated in the ovary, suggesting that ovarian oogenesis tends to be energized by aerobic metabolism. The TCA cycle may be used more for providing biosynthetic precursors and facilitating the transport of substrates between the mitochondrion and the cytoplasm rather than just as a source of ATP. Muscle tissue relies primarily on glycolysis for rapid energy production and may redistribute energy to the gonads, prioritizing the energy needs of the ovaries and contributing to the dynamic balance between reproduction and growth. This study provides insights into the molecular mechanisms of how environmental factors regulate fish reproduction, providing a theoretical basis and potential molecular targets for the regulation of reproduction and optimization of aquaculture environments. Full article
(This article belongs to the Section Molecular Biology)
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