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14 pages, 3029 KiB  
Article
In Vitro Bioactivity and Cytotoxicity Assessment of Two Root Canal Sealers
by Yicheng Ye, Sepanta Hosseinpour, Juan Wen and Ove A. Peters
Materials 2025, 18(15), 3717; https://doi.org/10.3390/ma18153717 (registering DOI) - 7 Aug 2025
Abstract
The development of bioactive materials in endodontics has advanced tissue regeneration by enhancing the biological responses of periradicular tissues. Recently, calcium silicate-based sealers have gained attention for their superior biological properties, including biocompatibility, osteoconductivity, and cementogenic potential. This study aimed to evaluate the [...] Read more.
The development of bioactive materials in endodontics has advanced tissue regeneration by enhancing the biological responses of periradicular tissues. Recently, calcium silicate-based sealers have gained attention for their superior biological properties, including biocompatibility, osteoconductivity, and cementogenic potential. This study aimed to evaluate the cytotoxicity, biocompatibility, and bioactivity of EndoSequence BC Sealer (ES BC) and AH Plus Bioceramic Sealer (AHP BC) using human periodontal ligament stromal cells (hPDLSCs). Biocompatibility was assessed using MTT, Live/Dead, and wound healing assays. ES BC and AHP BC demonstrated significantly higher cell viability and proliferation compared to AH Plus used as a control. Gene expression analysis via real-time quantitative PCR demonstrated that ES BC, especially in set form, significantly upregulated osteogenic markers—alkaline phosphatase (2.49 ± 0.10, p < 0.01), runt-related transcription factor 2 (2.33 ± 0.13), and collagen type I alpha 1 chain (2.85 ± 0.40, p < 0.001)—more than cementogenic markers (cementum protein 1, cementum attachment protein, and cementum protein 23). This differential response may reflect the fibroblast-dominant nature of hPDLSCs, which contain limited cementoblast-like cells. This study supports the superior biocompatibility and regenerative capacity of ES BC and AHP BC compared to AH Plus. While in vitro models provide foundational insights, advanced ex vivo approaches are crucial for translating findings to clinical practice. Full article
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19 pages, 4153 KiB  
Article
Whole-Genome Resequencing Analysis of Athletic Traits in Grassland-Thoroughbred
by Wenqi Ding, Wendian Gong, Tugeqin Bou, Lin Shi, Yanan Lin, Xiaoyuan Shi, Zheng Li, Huize Wu, Manglai Dugarjaviin and Dongyi Bai
Animals 2025, 15(15), 2323; https://doi.org/10.3390/ani15152323 (registering DOI) - 7 Aug 2025
Abstract
Speed is not only the primary objective of racehorse breeding but also a crucial indicator for evaluating racehorse performance. This study investigates a newly developed racehorse breed in China. Through whole-genome resequencing, we selected 60 offspring obtained from the crossbreeding of Thoroughbred horses [...] Read more.
Speed is not only the primary objective of racehorse breeding but also a crucial indicator for evaluating racehorse performance. This study investigates a newly developed racehorse breed in China. Through whole-genome resequencing, we selected 60 offspring obtained from the crossbreeding of Thoroughbred horses and Xilingol horses for this study. This breed is tentatively named “Grassland-Thoroughbred”, and the samples were divided into two groups based on racing ability: 30 racehorses and 30 non-racehorses. Based on whole-genome sequencing data, the study achieved an average sequencing depth of 25.63×. The analysis revealed strong selection pressure on chromosomes (Chr) 1 and 3. Selection signals were detected using methods such as the nucleotide diversity ratio (π ratio), integrated haplotype score (iHS), fixation index (Fst), and cross-population extended haplotype homozygosity (XP-EHH). Regions ranked in the top 5% by at least three methods were designated as candidate regions. This approach detected 215 candidate genes. Additionally, the Fst method was employed to detect Indels, and the top 1% regions detected were considered candidate regions, covering 661 candidate genes. Functional enrichment analysis of the candidate genes suggests that pathways related to immune regulation, neural signal transmission, muscle contraction, and energy metabolism may significantly influence differences in performance. Among these identified genes, PPARGC1A, FOXO1, SGCD, FOXP2, PRKG1, SLC25A15, CKMT2, and TRAP1 play crucial roles in muscle function, metabolism, sensory perception, and neurobiology, indicating their key significance in shaping racehorse phenotypes. This study not only enhances understanding of the molecular mechanisms underlying racehorse speed but also provides essential theoretical and practical references for the molecular breeding of Grassland-Thoroughbreds. Full article
(This article belongs to the Section Animal Genetics and Genomics)
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20 pages, 3576 KiB  
Article
Urban Wetland Sediments in Yangzhou: Physicochemical Properties, Microbial Communities, and Functional Associations
by Dongmei He, Liwen Li, Runyang Zhou, Sumei Qiu, Wei Xing and Yingdan Yuan
Microorganisms 2025, 13(8), 1843; https://doi.org/10.3390/microorganisms13081843 - 7 Aug 2025
Abstract
Urban wetlands play a crucial role in maintaining ecological balance, carbon sequestration, and water purification. Sediments are key carriers for wetlands to store elements such as carbon, nitrogen, and phosphorus in the aquatic environment. This study analyzed different sediment layers of seven wetlands [...] Read more.
Urban wetlands play a crucial role in maintaining ecological balance, carbon sequestration, and water purification. Sediments are key carriers for wetlands to store elements such as carbon, nitrogen, and phosphorus in the aquatic environment. This study analyzed different sediment layers of seven wetlands in Yangzhou, aiming to explore the relationship between physicochemical factors and microbial communities in wetland sediments, as well as to predict the functions of microbial communities. Functional prediction of microbial communities was conducted based on amplicon sequencing analysis, and the neutral community model was used to determine the formation and evolution process of microbial communities. The results showed that in three wetlands, namely Zhuyu Bay (ZYW), Luyang Lake (LYH), and Runyang Wetland (RYSD), the contents of carbon components (total carbon, total soluble carbon, microbial biomass carbon) in the 0–20 cm sediment layer were higher, while the carbon component contents in Baoying Lake (BYH) showed the opposite trend. Among them, the contents of total nitrogen, alkali-hydrolyzable nitrogen, total phosphorus, available phosphorus, total potassium, and available potassium in the 0–20 cm sediment layer of Runyang Wetland (RYSD) were significantly the highest. This indicates that in Runyang Wetland (RYSD), the 0–20 cm layer has more abundant carbon components and mineral nutrients compared to the 20–40 cm layer. Among the seven wetlands, it was found that the content of total potassium was all greater than 10 g/kg, which was much higher than the contents of total phosphorus and total nitrogen. Analysis of microbial communities revealed that the dominant archaeal phyla were Thaumarchaeota and Euryarchaeota, and the dominant bacterial phyla were Proteobacteria and Acidobacteria. The distribution of functional genes was mainly concentrated in Zhuyu Bay (ZYW) and Luyang Lake (LYH). Zhuyu Bay Wetland (ZYW) had potential advantages in light utilization function, and Luyang Lake (LYH) had potential advantages in carbon and nitrogen cycle functions. The assembly process of the archaeal community was mainly affected by stochastic processes, while the bacterial community was mainly affected by deterministic processes. However, water content, total phosphorus, and available potassium all had strong correlations with both archaeal and bacterial communities. The research results preliminarily reveal the connections between the physicochemical properties of sediments, microbial communities, and their potential functions in Yangzhou urban wetlands, providing an important scientific basis for the protection and management of wetland ecosystems. Full article
(This article belongs to the Section Environmental Microbiology)
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20 pages, 5610 KiB  
Article
The Gut Microbial Adaptation of Wild Goitered Gazelles Under Antibiotic Pressure in the Qaidam Basin
by Qing Zhao, Yiran Wang, Jingqing Ma and Wen Qin
Microorganisms 2025, 13(8), 1842; https://doi.org/10.3390/microorganisms13081842 - 7 Aug 2025
Abstract
Gut microbiota plays a vital role in host resilience but may be disrupted under environmental antibiotic pressure. The goitered gazelle (Gazella subgutturosa), a keystone ungulate in the Qaidam Basin, is crucial for ecosystem stability. We aimed to investigate how this species [...] Read more.
Gut microbiota plays a vital role in host resilience but may be disrupted under environmental antibiotic pressure. The goitered gazelle (Gazella subgutturosa), a keystone ungulate in the Qaidam Basin, is crucial for ecosystem stability. We aimed to investigate how this species responds to antibiotic pressure through gut microbial adaptation. Using 16S rRNA sequencing and weighted gene co-expression network analysis (WGCNA) on fecal and soil samples from six regions, we identified 18 microbial modules, of which three were strongly associated with antibiotics (|r| ≥ 0.75, p < 0.05). Gut microbial α-diversity was lowest in the antibiotic-rich, vegetation-poor TGL region and highest in XRH, where diverse vegetation appeared to buffer antibiotic impact. Antibiotic pressure can reshape gut microbial communities, exerting both adaptive benefits and adverse effects. High-quality habitats may alleviate the negative impacts of antibiotic pressure. Full article
(This article belongs to the Section Gut Microbiota)
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23 pages, 2328 KiB  
Article
Novel Insights into T-Cell Exhaustion and Cancer Biomarkers in PDAC Using ScRNA-Seq
by Muhammad Usman Saleem, Hammad Ali Sajid, Muhammad Waqar Arshad, Alejandro Omar Rivera Torres, Muhammad Imran Shabbir and Sunil Kumar Rai
Biology 2025, 14(8), 1015; https://doi.org/10.3390/biology14081015 - 7 Aug 2025
Abstract
One of the aggressive and lethal cancers, pancreatic ductal adenocarcinoma (PDAC) is characterized by poor prognosis and resistance to conventional treatments. Moreover, the tumor immune microenvironment (TIME) plays a crucial role in the progression and therapeutic resistance of PDAC. It is associated with [...] Read more.
One of the aggressive and lethal cancers, pancreatic ductal adenocarcinoma (PDAC) is characterized by poor prognosis and resistance to conventional treatments. Moreover, the tumor immune microenvironment (TIME) plays a crucial role in the progression and therapeutic resistance of PDAC. It is associated with T-cell exhaustion, leading to the progressive loss of T-cell functions with an impaired ability to kill tumor cells. Therefore, this study employed single-cell RNA sequencing (scRNA-seq) analysis of a publicly available human PDAC dataset, with cells isolated from the primary tumor and adjacent normal tissues, identifying upregulated genes of T-cells and cancer cells in two groups (“cancer cells_vs_all-PDAC” and “cancer-PDAC_vs_all-normal”). Common and unique markers of cancer cells from both groups were identified. The Reactome pathways of cancer and T-cells were selected, while the genes implicated in those pathways were used to perform PPI analysis, revealing the hub genes of cancer and T-cells. The gene expression validation of cancer and T-cells hub-genes was performed using GEPIA2 and TISCH2, while the overall survival analysis of cancer cells hub-genes was performed using GEPIA2. Conclusively, this study unraveled 16 novel markers of cancer and T-cells, providing the groundwork for future research into the immune landscape of PDAC, particularly T-cell exhaustion. However, further clinical studies are needed to validate these novel markers as potential therapeutic targets in PDAC patients. Full article
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21 pages, 3405 KiB  
Article
Allelic Variation of Helicobacter pylori vacA Gene and Its Association with Gastric Pathologies in Clinical Samples Collected in Jordan
by Mamoon M. Al-Hyassat, Hala I. Al-Daghistani, Lubna F. Abu-Niaaj, Sima Zein and Talal Al-Qaisi
Microorganisms 2025, 13(8), 1841; https://doi.org/10.3390/microorganisms13081841 - 7 Aug 2025
Abstract
Helicobacter pylori is a well-established causative agent of gastritis, peptic ulcers, gastric adenocarcinoma, and primary gastric lymphoma. It colonizes the human stomach and expresses numerous virulent factors that influence disease progression. Among these factors is the cytotoxin vacA gene, which encodes the vacuolating [...] Read more.
Helicobacter pylori is a well-established causative agent of gastritis, peptic ulcers, gastric adenocarcinoma, and primary gastric lymphoma. It colonizes the human stomach and expresses numerous virulent factors that influence disease progression. Among these factors is the cytotoxin vacA gene, which encodes the vacuolating capacity of the cytotoxin and plays a key role in the bacterium’s pathogenic potential. This study investigated the allelic diversity of the vacA among H. pylori strains infecting patients in Jordan with various gastric conditions and examined potential associations between vacA s-and m- genotypes, histopathological and endoscopic findings, and the development of gastric diseases. Gastric biopsies were collected from 106 patients at two hospitals in Jordan who underwent endoscopic examination. The collected biopsies for each patient were subjected to histopathological assessment, urease detection using the Rapid Urease Test (RUT), a diagnostic test for H. pylori, and molecular detection of the vacA gene and its s and m alleles. The histopathology reports indicated that 83 of 106 patients exhibited gastric disorders, of which 81 samples showed features associated with H. pylori infection. The RUT was positive in 76 of 106 with an accuracy of 93.8%. Real-time polymerase chain reaction (RT-PCR) targeting the 16S rRNA gene confirmed the presence of H. pylori in 79 of 81 histologically diagnosed cases as infected (97.5%), while the vacA gene was detected only in 75 samples (~95%). To explore genetic diversity, PCR-amplified fragments underwent sequence analysis of the vacA gene. The m-allele was detected in 58 samples (73%), the s-allele was detected in 45 (57%), while both alleles were not detected in 13% of samples. The predominant genotype combination among Jordanians was vacA s2/m2 (50%), significantly linked to mild chronic gastritis, followed by s1/m2 (35%) and s1/m1 (11.8%) which are linked to severe gastric conditions including malignancies. Age-and gender-related differences in vacA genotype were observed with less virulent s2m2 and s1m2 genotypes predominating in younger adults specially males, while the more virulent m1 genotypes were found exclusively in females and middle-aged patients. Genomic sequencing revealed extensive diversity within H. pylori, likely reflecting its long-standing co-evolution with human hosts in Jordan. This genetic variability plays a key role in modulating virulence and influencing clinical outcomes. Comprehensive characterization of vacA genotypic variations through whole-genome sequencing is essential to enhance diagnostic precision, strengthen epidemiological surveillance, and inform targeted therapeutic strategies. While this study highlights the significance of the vacA m and s alleles, future research is recommended in order to investigate the other vacA allelic variations, such as the i, d, and c alleles, to achieve a more comprehensive understanding of H. pylori pathogenicity and associated disease severity across different strains. These investigations will be crucial for improving diagnostic accuracy and guiding the development of targeted therapeutic strategies. Full article
(This article belongs to the Special Issue Helicobacter pylori Infection: Detection and Novel Treatment)
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15 pages, 8949 KiB  
Article
Protein Expression of TXNIP in the Dopaminergic Neurons of Subjects with Parkinson’s Disease: Evidence from a Pilot Study
by Francesca A. Schillaci, Giuseppe Lanza, Maria Grazia Salluzzo, Raffaele Ferri and Michele Salemi
Life 2025, 15(8), 1252; https://doi.org/10.3390/life15081252 - 7 Aug 2025
Abstract
Parkinson’s disease (PD) is a progressive, multisystemic α-synucleinopathy, recognized as the second most prevalent neurodegenerative disorder globally. Its neuropathology is characterized by the degeneration of dopaminergic neurons, particularly in the substantia nigra pars compacta (SNpc), and the intraneuronal accumulation of α-synuclein-forming Lewy bodies. [...] Read more.
Parkinson’s disease (PD) is a progressive, multisystemic α-synucleinopathy, recognized as the second most prevalent neurodegenerative disorder globally. Its neuropathology is characterized by the degeneration of dopaminergic neurons, particularly in the substantia nigra pars compacta (SNpc), and the intraneuronal accumulation of α-synuclein-forming Lewy bodies. Oxidative stress is a key contributor to PD pathogenesis. Thioredoxin-interacting protein (TXNIP) is a crucial regulator of cellular redox balance, inhibiting the antioxidant function of thioredoxin. This pilot study aimed to investigate the protein expression and localization of TXNIP in the SNpc of PD patients compared to healthy controls. We performed immunohistochemical analyses on 12 post-mortem human brain sections (formalin-fixed, paraffin-embedded) from six subjects with PD and six healthy controls. The study was performed on PD subjects with Braak stage 6. Our findings revealed that in control samples, TXNIP protein was distinctly and closely associated with neuromelanin (NM) pigment within the cytoplasm of SNpc dopaminergic neurons. Conversely, in PD samples, there was a markedly weak cytoplasmic expression of TXNIP, and critically, this association with NM pigment was absent. Furthermore, PD samples exhibited a significant reduction in both dopaminergic neurons and NM content, consistent with advanced disease. These findings, which mirror previous transcriptomic data showing TXNIP gene under-expression in the same subjects, suggest that altered TXNIP expression and localization in SNpc dopaminergic neurons are features of late-stage PD, potentially reflecting neuronal dysfunction and loss. Full article
(This article belongs to the Special Issue Regulation of Cellular Signaling Pathways in the Metabolic Syndrome)
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18 pages, 3146 KiB  
Article
Quorum-Sensing C12-HSL Drives Antibiotic Resistance Plasmid Transfer via Membrane Remodeling, Oxidative Stress, and RpoS-RMF Crosstalk
by Yang Yang, Ziyan Wu, Li’e Zhu, Zixin Han, Junpeng Li, Qiaoqiao Fang and Guoqiang Zhu
Microorganisms 2025, 13(8), 1837; https://doi.org/10.3390/microorganisms13081837 - 6 Aug 2025
Abstract
Antibiotic misuse accelerates resistance dissemination via plasmid conjugation, but quorum sensing (QS) regulatory mechanisms remain undefined. Using Escherichia coli (E. coli) MG1655 conjugation models (RP4-7/EC600 plasmids), we demonstrate that long-chain acyl-homoserine lactones (C10/C12-HSL) enhance transfer frequency by up to 7.7-fold (200 [...] Read more.
Antibiotic misuse accelerates resistance dissemination via plasmid conjugation, but quorum sensing (QS) regulatory mechanisms remain undefined. Using Escherichia coli (E. coli) MG1655 conjugation models (RP4-7/EC600 plasmids), we demonstrate that long-chain acyl-homoserine lactones (C10/C12-HSL) enhance transfer frequency by up to 7.7-fold (200 μM C12-HSL; p < 0.001), while quorum-quenching by sub-inhibitory vanillin suppressed this effect by 95% (p < 0.0001). C12-HSL compromised membrane integrity via ompF upregulation (4-fold; p < 0.01) and conjugative pore assembly (trbBp upregulated by 1.38-fold; p < 0.05), coinciding with ROS accumulation (1.5-fold; p < 0.0001) and SOS response activation (recA upregulated by 1.68-fold; p < 0.001). Crucially, rpoS and rmf deletion mutants reduced conjugation by 65.5% and 55.8%, respectively (p < 0.001), exhibiting attenuated membrane permeability (≤65.5% reduced NPN influx; p < 0.0001), suppressed ROS (≤54% downregulated; p < 0.0001), and abolished transcriptional induction of conjugation/stress genes. Reciprocal RpoS–RMF (ribosomal hibernation factor) crosstalk was essential for AHL responsiveness, with deletions mutually suppressing expression (≤65.9% downregulated; p < 0.05). We establish a hierarchical mechanism wherein long-chain AHLs drive resistance dissemination through integrated membrane restructuring, stress adaptation, and RpoS–RMF-mediated genetic plasticity, positioning QS signaling as a viable target for curbing resistance spread. Full article
(This article belongs to the Section Antimicrobial Agents and Resistance)
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23 pages, 8569 KiB  
Article
Evidential K-Nearest Neighbors with Cognitive-Inspired Feature Selection for High-Dimensional Data
by Yawen Liu, Yang Zhang, Xudong Wang and Xinyuan Qu
Big Data Cogn. Comput. 2025, 9(8), 202; https://doi.org/10.3390/bdcc9080202 - 6 Aug 2025
Abstract
The Evidential K-Nearest Neighbor (EK-NN) classifier has demonstrated robustness in handling incomplete and uncertain data; however, its application in high-dimensional big data for feature selection, such as genomic datasets with tens of thousands of gene features, remains underexplored. Our proposed Granular–Elastic Evidential K-Nearest [...] Read more.
The Evidential K-Nearest Neighbor (EK-NN) classifier has demonstrated robustness in handling incomplete and uncertain data; however, its application in high-dimensional big data for feature selection, such as genomic datasets with tens of thousands of gene features, remains underexplored. Our proposed Granular–Elastic Evidential K-Nearest Neighbor (GEK-NN) approach addresses this gap. In the context of big data, GEK-NN integrates an Elastic Net within the Genetic Algorithm’s fitness function to efficiently sift through vast amounts of data, identifying relevant feature subsets. This process mimics human cognitive behavior of filtering and refining information, similar to concepts in cognitive computing. A granularity metric is further employed to optimize subset size, maximizing its impact. GEK-NN consists of two crucial phases. Initially, an Elastic Net-based feature evaluation is conducted to pinpoint relevant features from the high-dimensional data. Subsequently, granularity-based optimization refines the subset size, adapting to the complexity of big data. Before applying to genomic big data, experiments on UCI datasets demonstrated the feasibility and effectiveness of GEK-NN. By using an Evidence Theory framework, GEK-NN overcomes feature-selection challenges in both low-dimensional UCI datasets and high-dimensional genomic big data, significantly enhancing pattern recognition and classification accuracy. Comparative analyses with existing EK-NN feature-selection methods, using both UCI and high-dimensional gene datasets, underscore GEK-NN’s superiority in handling big data for feature selection and classification. These results indicate that GEK-NN not only enriches EK-NN applications but also offers a cognitive-inspired solution for complex gene data analysis, effectively tackling high-dimensional feature-selection challenges in the realm of big data. Full article
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11 pages, 811 KiB  
Article
Activity Expression and Property Analysis of Codon-Optimized Polyphenol Oxidase from Camellia sinensis in Pichia pastoris KM71
by Xin Zhang, Yong-Quan Xu, Jun-Feng Yin and Chun Zou
Foods 2025, 14(15), 2749; https://doi.org/10.3390/foods14152749 - 6 Aug 2025
Abstract
Tea polyphenol oxidase (CsPPO) is a crucial enzyme involved in the production of tea and tea products. However, the recombinant expression of CsPPO in microorganisms is often hindered by challenges such as inclusion body formation and extremely low enzyme activity. In this study, [...] Read more.
Tea polyphenol oxidase (CsPPO) is a crucial enzyme involved in the production of tea and tea products. However, the recombinant expression of CsPPO in microorganisms is often hindered by challenges such as inclusion body formation and extremely low enzyme activity. In this study, the CsPPO gene (1800 bp) from Camellia sinensis cv. Yihongzao was cloned and 14.5% of its codons were optimized for Pichia pastoris expression. Compared to pre-optimization, codon optimization significantly enhanced CsPPO production in P. pastoris KM71, yielding a 42.89-fold increase in enzyme activity (1286.67 U/mL). The optimal temperature and pH for recombinant CsPPO were determined to be 40 °C and 5.5, respectively. This study demonstrates that codon optimization effectively improves the expression of plant-derived enzymes such as CsPPO in eukaryotic expression systems. Future research should explore the long-term stability of recombinant CsPPO and its potential applications in tea fermentation processes and functional food development. Full article
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16 pages, 1298 KiB  
Article
Genetic Effects of Chicken Pre-miR-3528 SNP on Growth Performance, Meat Quality Traits, and Serum Enzyme Activities
by Jianzhou Shi, Jinbing Zhao, Bingxue Dong, Na Li, Lunguang Yao and Guirong Sun
Animals 2025, 15(15), 2300; https://doi.org/10.3390/ani15152300 - 6 Aug 2025
Abstract
The aim was to investigate the genetic effects of a SNP located in the precursor region of gga-miR-3528. (1) Single-nucleotide polymorphisms within precursor regions of microRNAs play crucial biological roles. (2) Utilizing a Gushi–Anka F2 resource population (n = 860), [...] Read more.
The aim was to investigate the genetic effects of a SNP located in the precursor region of gga-miR-3528. (1) Single-nucleotide polymorphisms within precursor regions of microRNAs play crucial biological roles. (2) Utilizing a Gushi–Anka F2 resource population (n = 860), we screened and validated miRNA SNPs. A SNP mutation in the miR-3528 precursor region was identified. Specific primers were designed to amplify the polymorphic fragment. Genotyping was performed for this individual SNP across the population, using the MassArray system. Association analyses were conducted between this SNP and chicken growth and body measurement traits, carcass traits, meat quality traits, and serum enzyme activities. (3) The rs14098602 (+12 bp A > G) was identified within the precursor region of gga-miR-3528. Significant associations (p < 0.05) were observed between this SNP and chicken growth traits (body weight at the age of 0 day, body weight at the age of 2 weeks, and body weight at the age of 4 weeks), carcass traits (evisceration weight), meat quality traits (subcutaneous fat rate and pectoral muscle density), and serum enzyme activities (total protein, albumin, globulin, cholinesterase, and lactate dehydrogenase). (4) These findings suggest that the polymorphism at rs14098602 may influence chicken growth, meat quality, and serum biochemical indices, through specific mechanisms. The gga-miR-3528 gene likely plays an important role in chicken development. Therefore, this SNP can serve as a molecular marker for genetic breeding and auxiliary selection of growth-related traits, facilitating the rapid establishment of elite chicken populations with superior genetic resources. Full article
(This article belongs to the Section Poultry)
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19 pages, 3503 KiB  
Article
Discovery of Hub Genes Involved in Seed Development and Lipid Biosynthesis in Sea Buckthorn (Hippophae rhamnoides L.) Using UID Transcriptome Sequencing
by Siyang Zhao, Chengjiang Ruan, Alexey A. Dmitriev and Hyun Uk Kim
Plants 2025, 14(15), 2436; https://doi.org/10.3390/plants14152436 - 6 Aug 2025
Abstract
Sea buckthorn is a vital woody oil species valued for its role in soil conservation and its bioactive seed oil, which is rich in unsaturated fatty acids and other compounds. However, low seed oil content and small seed size are the main bottlenecks [...] Read more.
Sea buckthorn is a vital woody oil species valued for its role in soil conservation and its bioactive seed oil, which is rich in unsaturated fatty acids and other compounds. However, low seed oil content and small seed size are the main bottlenecks restricting the development and utilization of sea buckthorn. In this study, we tested the seed oil content and seed size of 12 sea buckthorn cultivars and identified the key genes and transcription factors involved in seed development and lipid biosynthesis via the integration of UID RNA-seq (Unique Identifiers, UID), WGCNA (weighted gene co-expression network analysis) and qRT-PCR (quantitative real-time PCR) analysis. The results revealed five cultivars (CY02, CY11, CY201309, CY18, CY21) with significantly higher oil contents and five cultivars (CY10, CY201309, CY18, CY21, CY27) with significantly heavier seeds. A total of 10,873 genes were significantly differentially expressed between the S1 and S2 seed developmental stages of the 12 cultivars. WGCNA was used to identify five modules related to seed oil content and seed weight/size, and 417 candidate genes were screened from these modules. Among them, multiple hub genes and transcription factors were identified; for instance, ATP synthase, ATP synthase subunit D and Acyl carrier protein 1 were related to seed development; plastid–lipid-associated protein, acyltransferase-like protein, and glycerol-3-phosphate 2-O-acyltransferase 6 were involved in lipid biosynthesis; and transcription factors DOF1.2, BHLH137 and ERF4 were associated with seed enlargement and development. These findings provide crucial insights into the genetic regulation of seed traits in sea buckthorn, offering targets for future breeding efforts aimed at improving oil yield and quality. Full article
(This article belongs to the Special Issue Molecular Regulation of Seed Development and Germination)
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16 pages, 1898 KiB  
Article
Screening of qPCR Reference Genes in Quinoa Under Cold, Heat, and Drought Gradient Stress
by Qiuwei Lu, Xueying Wang, Suxuan Dong, Jinghan Fu, Yiqing Lin, Ying Zhang, Bo Zhao and Fuye Guo
Plants 2025, 14(15), 2434; https://doi.org/10.3390/plants14152434 - 6 Aug 2025
Abstract
Quinoa (Chenopodium quinoa), a stress-tolerant pseudocereal ideal for studying abiotic stress responses, was used to systematically identify optimal reference genes for qPCR normalization under gradient stresses: low temperatures (LT group: −2 °C to −10 °C), heat (HT group: 39° C to [...] Read more.
Quinoa (Chenopodium quinoa), a stress-tolerant pseudocereal ideal for studying abiotic stress responses, was used to systematically identify optimal reference genes for qPCR normalization under gradient stresses: low temperatures (LT group: −2 °C to −10 °C), heat (HT group: 39° C to 45 °C), and drought (DR group: 7 to 13 days). Through multi-algorithm evaluation (GeNorm, NormFinder, BestKeeper, the ΔCt method, and RefFinder) of eleven candidates, condition-specific optimal genes were established as ACT16 (Actin), SAL92 (IT4 phosphatase-associated protein), SSU32 (Ssu72-like family protein), and TSB05 (Tryptophan synthase beta-subunit 2) for the LT group; ACT16 and NRP13 (Asparagine-rich protein) for the HT group; and ACT16, SKP27 (S-phase kinase), and NRP13 for the DR group, with ACT16, NRP13, WLIM96 (LIM domain-containing protein), SSU32, SKP27, SAL92, and UBC22 (ubiquitin-conjugating enzyme E2) demonstrating cross-stress stability (global group). DHDPS96 (dihydrodipicolinate synthase) and EF03 (translation elongation factor) showed minimal stability. Validation using stress-responsive markers—COR72 (LT), HSP44 (HT), COR413-PM (LT), and DREB12 (DR)—confirmed reliability; COR72 and COR413-PM exhibited oscillatory cold response patterns, HSP44 peaked at 43 °C before declining, and DREB12 showed progressive drought-induced upregulation. Crucially, normalization with unstable genes (DHDPS96 and EF03) distorted expression profiles. This work provides validated reference standards for quinoa transcriptomics under abiotic stresses. Full article
(This article belongs to the Section Plant Response to Abiotic Stress and Climate Change)
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18 pages, 617 KiB  
Article
GNR: Genetic-Embedded Nuclear Reaction Optimization with F-Score Filter for Gene Selection in Cancer Classification
by Shahad Alkamli and Hala Alshamlan
Int. J. Mol. Sci. 2025, 26(15), 7587; https://doi.org/10.3390/ijms26157587 - 6 Aug 2025
Abstract
The classification of cancer based on gene expression profiles is a central challenge in precision oncology due to the high dimensionality and low sample size inherent in microarray datasets. Effective gene selection is crucial for improving classification accuracy while minimizing computational overhead and [...] Read more.
The classification of cancer based on gene expression profiles is a central challenge in precision oncology due to the high dimensionality and low sample size inherent in microarray datasets. Effective gene selection is crucial for improving classification accuracy while minimizing computational overhead and model complexity. This study introduces Genetic-Embedded Nuclear Reaction Optimization (GNR), a novel hybrid metaheuristic that enhances the conventional Nuclear Reaction Optimization (NRO) algorithm by embedding a genetic uniform crossover mechanism into its fusion phase. The proposed algorithm leverages a two-stage process: an initial F-score filtering step to reduce dimensionality, followed by GNR-driven optimization to identify compact, informative gene subsets. Evaluations were conducted on six widely used microarray cancer datasets, with Support Vector Machines (SVM) employed as classifiers and performance assessed via Leave-One-Out Cross-Validation (LOOCV). Results show that GNR consistently outperforms the original NRO and several benchmark hybrid algorithms, achieving 100% classification accuracy with significantly smaller gene subsets across all datasets. These findings confirm the efficacy of the genetic-embedded fusion strategy in enhancing local exploitation while preserving the global search capabilities of NRO, thereby offering a robust and interpretable approach for gene selection in cancer classification. Full article
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24 pages, 10588 KiB  
Article
Genome-Wide Identification, Evolution, and Expression Patterns of the Fructose-1,6-Bisphosphatase Gene Family in Saccharum Species
by Chunyan Tian, Xiuting Hua, Peifang Zhao, Chunjia Li, Xujuan Li, Hongbo Liu and Xinlong Liu
Plants 2025, 14(15), 2433; https://doi.org/10.3390/plants14152433 - 6 Aug 2025
Abstract
Fructose-1,6-bisphosphatase (FBP) is a crucial regulatory enzyme in sucrose synthesis and photosynthetic carbon assimilation, functioning through two distinct isoforms: cytosolic FBP (cyFBP) and chloroplastic FBP (cpFBP). However, the identification and functional characterization of FBP genes in Saccharum remains limited. In this study, we [...] Read more.
Fructose-1,6-bisphosphatase (FBP) is a crucial regulatory enzyme in sucrose synthesis and photosynthetic carbon assimilation, functioning through two distinct isoforms: cytosolic FBP (cyFBP) and chloroplastic FBP (cpFBP). However, the identification and functional characterization of FBP genes in Saccharum remains limited. In this study, we conducted a systematic identification and comparative genomics analyses of FBPs in three Saccharum species. We further examined their expression patterns across leaf developmental zones, spatiotemporal profiles, and responses to diurnal rhythms and hormonal treatments. Our analysis identified 95 FBP genes, including 44 cyFBPs and 51 cpFBPs. Comparative analyses revealed significant divergence in physicochemical properties, gene structures, and motif compositions between the two isoforms. Expression profiling indicated that both cyFBPs and cpFBPs were predominantly expressed in leaves, particularly in maturing and mature zones. During diurnal cycles, their expression peaked around the night–day transition, with cpFBPs exhibiting earlier peaks than cyFBPs. FBP genes in Saccharum spontaneum displayed greater diurnal sensitivity than those in Saccharum officinarum. Hormonal treatments further revealed significant regulatory divergence in FBP genes, both between isoforms and across species. Notably, cyFBP_2 and cpFBP_2 members consistently exhibited higher expression levels across all datasets, suggesting their pivotal roles in sugarcane physiology. These findings not only identify potential target genes for enhancing sucrose accumulation, but also highlight the breeding value of S. spontaneum and S. officinarum in sugarcane breeding. Full article
(This article belongs to the Section Plant Genetics, Genomics and Biotechnology)
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