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21 pages, 1784 KiB  
Article
Toxic Threats from the Fern Pteridium aquilinum: A Multidisciplinary Case Study in Northern Spain
by L. María Sierra, Isabel Feito, Mª Lucía Rodríguez, Ana Velázquez, Alejandra Cué, Jaime San-Juan-Guardado, Marta Martín, Darío López, Alexis E. Peña, Elena Canga, Guillermo Ramos, Juan Majada, José Manuel Alvarez and Helena Fernández
Int. J. Mol. Sci. 2025, 26(15), 7157; https://doi.org/10.3390/ijms26157157 - 24 Jul 2025
Viewed by 191
Abstract
Pteridium aquilinum (bracken fern) poses a global threat to biodiversity and to the health of both animals and humans due to its toxic metabolites and aggressive ecological expansion. In northern Spain, particularly in regions of intensive livestock farming, these risks may be exacerbated, [...] Read more.
Pteridium aquilinum (bracken fern) poses a global threat to biodiversity and to the health of both animals and humans due to its toxic metabolites and aggressive ecological expansion. In northern Spain, particularly in regions of intensive livestock farming, these risks may be exacerbated, calling for urgent assessment and monitoring strategies. In this study, we implemented a multidisciplinary approach to evaluate the toxicological and ecological relevance of P. aquilinum through four key actions: (a) quantification of pterosins A and B in young fronds (croziers) using ultra-high-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS); (b) analysis of in vivo genotoxicity of aqueous extracts using Drosophila melanogaster as a model organism; (c) a large-scale survey of local livestock farmers to assess awareness and perceived impact of bracken; and (d) the development and field application of a drone-based mapping tool to assess the spatial distribution of the species at the regional level. Our results confirm the consistent presence of pterosins A and B in croziers, with concentrations ranging from 0.17 to 2.20 mg/g dry weight for PtrB and 13.39 to 257 µg/g for PtrA. Both metabolite concentrations and genotoxicity levels were found to correlate with latitude and, importantly, with each other. All tested samples exhibited genotoxic activity, with notable differences among them. The farmer survey (n = 212) revealed that only 50% of respondents were aware of the toxic risks posed by bracken, indicating a need for targeted outreach. The drone-assisted mapping approach proved to be a promising tool for identifying bracken-dominated areas and provides a scalable foundation for future ecological monitoring and land management strategies. Altogether, our findings emphasize that P. aquilinum is not merely a local concern but a globally relevant toxic species whose monitoring and control demand coordinated scientific and policy-based efforts. Full article
(This article belongs to the Special Issue The Transcendental World of Plant Toxic Compounds)
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14 pages, 3796 KiB  
Article
Preliminary Analysis of Placental DNA Methylation Profiles in Piglets with Extreme Birth Weight Variations
by Zhiyuan Zhang, Baohua Tan, Jiawei Su, Jiaming Xue, Liyao Xiao, Zicong Li, Linjun Hong, Gengyuan Cai and Ting Gu
Animals 2025, 15(15), 2168; https://doi.org/10.3390/ani15152168 - 23 Jul 2025
Viewed by 199
Abstract
Adequate birth weight is essential for animal survival and subsequent growth. However, the mechanism by which placental DNA methylation influences fetal growth remains incompletely understood. This study employed whole-genome bi-sulfite sequencing (WGBS) and RNA sequencing to analyze placental tissues from two weak piglets [...] Read more.
Adequate birth weight is essential for animal survival and subsequent growth. However, the mechanism by which placental DNA methylation influences fetal growth remains incompletely understood. This study employed whole-genome bi-sulfite sequencing (WGBS) and RNA sequencing to analyze placental tissues from two weak piglets and two normal piglets born to the same sow. Transcriptome analysis identified 1989 differentially expressed genes (DEGs) enriched in blood/immune processes. Additionally, differentially methylated regions linked to DEG repression were enriched in extracellular matrix (ECM) receptors and angiogenesis pathways. To investigate the role of DNA methylation in gene regulation, porcine trophoblast cells (PTr2) were treated with either DMSO (control) or the DNA methylation inhibitor 5-Aza-2′-deoxycytidine (5-Aza). Real-time quantitative PCR (RT-qPCR) analysis demonstrated significant upregulation of PACC1, SLC7A1, and PKP1 gene expression in the 5-Aza-treated group compared to controls (p < 0.05). Furthermore, methylation-specific PCR (MS-PCR) assays confirmed that the transcriptional activity of these genes is directly modulated by DNA methylation. These findings suggest that the dynamic regulation of DNA methylation in gene promoters may influence variations in placental morphology and birth weight in piglets, offering new insights into epigenetic regulation of fetal development, though larger studies are needed for validation. Full article
(This article belongs to the Special Issue Advances in Omics to Enhance Livestock Production)
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11 pages, 1607 KiB  
Article
Studies on the Emission of Volatile Organic Compounds from Selected Forest Mushrooms of the Genus Lactarius Using Proton-Transfer Reaction Mass Spectrometry
by Tomasz Wróblewski, Anna Kamińska and Agnieszka Włodarkiewicz
Molecules 2025, 30(14), 3000; https://doi.org/10.3390/molecules30143000 - 17 Jul 2025
Viewed by 265
Abstract
Forest mushrooms, due to their taste and smell, have been a component of people’s diets since the beginning of time. Unfortunately, there are many inedible or poisonous species of mushrooms that are similar to those that are eaten. For example, the highly valued [...] Read more.
Forest mushrooms, due to their taste and smell, have been a component of people’s diets since the beginning of time. Unfortunately, there are many inedible or poisonous species of mushrooms that are similar to those that are eaten. For example, the highly valued Boletus edulis is similar to the inedible bitter bolete and the poisonous bolete. In the case of mushrooms of the genus Lactarius, such similarities are demonstrated by the delicious tasting L. deliciosus, the inedible downy L. pubescens and the poisonous cottony L. torminosus. This study presents an attempt to classify these three species based on studies of the emission of volatile organic compounds from the volatile headspace using proton-transfer reaction mass spectrometry (PTR-MS). The conducted statistical tests, principal component analysis (PCA) and discriminant analysis revealed significant differences in the concentration of 20 selected protonated VOC molecules for the tested mushroom species. The clear advantages of the PTR-MS technique are that there is no need for special sample preparation and it has rapid measurement capability and high analytical sensitivity. This allows for a quick comparative analysis of VOCs, for example, from different species of forest mushrooms. Full article
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18 pages, 1568 KiB  
Article
Coupling of Temporal-Check-All-That-Apply and Nose-Space Analysis to Investigate the In Vivo Flavor Perception of Extra Virgin Olive Oil and Carriers’ Impact
by Danny Cliceri, Iuliia Khomenko, Franco Biasioli, Flavia Gasperi and Eugenio Aprea
Foods 2025, 14(13), 2343; https://doi.org/10.3390/foods14132343 - 1 Jul 2025
Viewed by 319
Abstract
The perceived quality of extra virgin olive oil (EVOO) arises from the multisensory integration of multimodal stimuli, primarily driven by non-volatile and volatile organic compounds (VOCs). Given that EVOO is frequently consumed in combination with other foods, cross-modal interactions, encompassing both internal and [...] Read more.
The perceived quality of extra virgin olive oil (EVOO) arises from the multisensory integration of multimodal stimuli, primarily driven by non-volatile and volatile organic compounds (VOCs). Given that EVOO is frequently consumed in combination with other foods, cross-modal interactions, encompassing both internal and external elements, play a crucial role in shaping its sensory perception. A more realistic representation of EVOO perception can be achieved by considering these cross-modal effects and their temporal dynamics. This study employed dynamic sensory and instrumental techniques to investigate the product-related mechanisms that influence EVOO flavor perception. Ten trained panelists (mean age = 41.5 years; 50% female) evaluated two EVOO samples under two consumption conditions: alone and accompanied by a solid carrier (bread or chickpeas). Temporal Check-All-That-Apply (TCATA) and nose-space analysis using Proton-Transfer-Reaction Time-of-Flight Mass Spectrometry (PTR-ToF-MS) were conducted simultaneously. Sensory descriptors and mass spectral peaks were analyzed through temporal curve indices (Area Under the Curve, Maximum Citation/Concentration, Time to Maximum), which were then used to construct multi-dimensional sensory and VOC release maps. Findings revealed that the composition and texture of the food carriers had a greater influence on temporal flavor perception than the variability in VOCs released by the different EVOO samples. These results underscore the importance of considering cross-modal sensory interactions when predicting EVOO flavor perception. The carriers modulated both the perception and VOC release, with effects dependent on their specific composition and texture. This methodological approach enabled a deeper understanding of the dynamic relationship between VOC release and EVOO sensory experience. Full article
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17 pages, 3134 KiB  
Article
Validation of Fiber-Dominant Expressing Gene Promoters in Populus trichocarpa
by Mengjie Guo, Ruxia Wang, Bo Wang, Wenjing Xu, Hui Hou, Hao Cheng, Yun Zhang, Chong Wang and Yuxiang Cheng
Plants 2025, 14(13), 1948; https://doi.org/10.3390/plants14131948 - 25 Jun 2025
Viewed by 547
Abstract
Wood is an important raw material for industrial applications. Its fiber-specific genetic modification provides an effective strategy to alter wood characteristics in tree breeding. Here, we performed a cross-analysis of previously reported single-cell RNA sequencing and the AspWood database during wood formation to [...] Read more.
Wood is an important raw material for industrial applications. Its fiber-specific genetic modification provides an effective strategy to alter wood characteristics in tree breeding. Here, we performed a cross-analysis of previously reported single-cell RNA sequencing and the AspWood database during wood formation to identify potential xylem fiber-dominant expressing genes in poplar. As a result, 32 candidate genes were obtained, and subsequently, we further examined the expression of these genes in fibers and/or vessels of stem secondary xylem using the laser capture microdissection technique and RT-qPCR. Analysis identified nine candidate genes, including PtrFLA12-2, PtrIRX12, PtrFLA12-6, PtrMYB52, PtrMYB103, PtrMAP70, PtrLRR-1, PtrKIFC2-3, and PtrNAC12. Next, we cloned the promoter regions of the nine candidate genes and created promoter::GUS transgenic poplars. Histochemical GUS staining was used to investigate the tissue expression activities of these gene promoters in transgenic poplars. In one month, transgenic plantlets grown in medium showed intensive GUS staining signals that were visible in the leaves and apical buds, suggesting substantial expression activities of these gene promoters in plantlets predominantly undergoing primary growth. In contrast, for three-month-old transgenic poplars in the greenhouse with predominantly developed secondary stem tissues, the promoters of seven of nine candidate genes, including PtrMYB103, PtrIRX12, and PtrMAP70, showed secondary xylem fiber-dominant GUS signals with considerable spatial specificity. Overall, this study presents xylem fiber-dominant promoters that are well-suited for specifically expressing genes of interest in wood fibers for forest tree breeding. Full article
(This article belongs to the Section Plant Physiology and Metabolism)
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13 pages, 657 KiB  
Article
Exhaled Breath Analysis in Lymphangioleiomyomatosis by Real-Time Proton Mass Spectrometry
by Malika Mustafina, Artemiy Silantyev, Marina Makarova, Aleksandr Suvorov, Alexander Chernyak, Zhanna Naumenko, Pavel Pakhomov, Ekaterina Pershina, Olga Suvorova, Anna Shmidt, Anastasia Gordeeva, Maria Vergun, Olesya Bahankova, Daria Gognieva, Aleksandra Bykova, Andrey Belevskiy, Sergey Avdeev, Vladimir Betelin and Philipp Kopylov
Int. J. Mol. Sci. 2025, 26(13), 6005; https://doi.org/10.3390/ijms26136005 - 23 Jun 2025
Viewed by 338
Abstract
Lymphangioleiomyomatosis (LAM) is a rare progressive disease that affects women of reproductive age and is characterized by cystic lung destruction, airflow obstruction, and lymphatic dysfunction. Current diagnostic methods are costly or lack sufficient specificity, highlighting the need for novel non-invasive approaches. Exhaled breath [...] Read more.
Lymphangioleiomyomatosis (LAM) is a rare progressive disease that affects women of reproductive age and is characterized by cystic lung destruction, airflow obstruction, and lymphatic dysfunction. Current diagnostic methods are costly or lack sufficient specificity, highlighting the need for novel non-invasive approaches. Exhaled breath analysis using real-time proton mass spectrometry (PTR-MS) presents a promising strategy for identifying disease-specific volatile organic compounds (VOCs). This cross-sectional study analyzed exhaled breath samples from 51 LAM patients and 51 age- and sex-matched healthy controls. PTR-time-of-flight mass spectrometry (PTR-TOF-MS) was employed to identify VOC signatures associated with LAM. Data preprocessing, feature selection, and statistical analyses were performed using machine learning models, including gradient boosting classifiers (XGBoost), to identify predictive biomarkers of LAM and its complications. We identified several VOCs as potential biomarkers of LAM, including m/z = 90.06 (lactic acid) and m/z = 113.13. VOCs predictive of disease complications included m/z = 49.00 (methanethiol), m/z = 48.04 (O-methylhydroxylamine), and m/z = 129.07, which correlated with pneumothorax, obstructive ventilation disorders, and radiological findings of lung cysts and bronchial narrowing. The classifier incorporating these biomarkers demonstrated high diagnostic accuracy (AUC = 0.922). This study provides the first evidence that exhaled breath VOC profiling can serve as a non-invasive additional tool for diagnosing LAM and predicting its complications. These findings warrant further validation in larger cohorts to refine biomarker specificity and explore their clinical applications in disease monitoring and personalized treatment strategies. Full article
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58 pages, 949 KiB  
Review
Excess Pollution from Vehicles—A Review and Outlook on Emission Controls, Testing, Malfunctions, Tampering, and Cheating
by Robin Smit, Alberto Ayala, Gerrit Kadijk and Pascal Buekenhoudt
Sustainability 2025, 17(12), 5362; https://doi.org/10.3390/su17125362 - 10 Jun 2025
Viewed by 1481
Abstract
Although the transition to electric vehicles (EVs) is well underway and expected to continue in global car markets, most vehicles on the world’s roads will be powered by internal combustion engine vehicles (ICEVs) and fossil fuels for the foreseeable future, possibly well past [...] Read more.
Although the transition to electric vehicles (EVs) is well underway and expected to continue in global car markets, most vehicles on the world’s roads will be powered by internal combustion engine vehicles (ICEVs) and fossil fuels for the foreseeable future, possibly well past 2050. Thus, good environmental performance and effective emission control of ICE vehicles will continue to be of paramount importance if the world is to achieve the stated air and climate pollution reduction goals. In this study, we review 228 publications and identify four main issues confronting these objectives: (1) cheating by vehicle manufacturers, (2) tampering by vehicle owners, (3) malfunctioning emission control systems, and (4) inadequate in-service emission programs. With progressively more stringent vehicle emission and fuel quality standards being implemented in all major markets, engine designs and emission control systems have become increasingly complex and sophisticated, creating opportunities for cheating and tampering. This is not a new phenomenon, with the first cases reported in the 1970s and continuing to happen today. Cheating appears not to be restricted to specific manufacturers or vehicle types. Suspicious real-world emissions behavior suggests that the use of defeat devices may be widespread. Defeat devices are primarily a concern with diesel vehicles, where emission control deactivation in real-world driving can lower manufacturing costs, improve fuel economy, reduce engine noise, improve vehicle performance, and extend refill intervals for diesel exhaust fluid, if present. Despite the financial penalties, undesired global attention, damage to brand reputation, a temporary drop in sales and stock value, and forced recalls, cheating may continue. Private vehicle owners resort to tampering to (1) improve performance and fuel efficiency; (2) avoid operating costs, including repairs; (3) increase the resale value of the vehicle (i.e., odometer tampering); or (4) simply to rebel against established norms. Tampering and cheating in the commercial freight sector also mean undercutting law-abiding operators, gaining unfair economic advantage, and posing excess harm to the environment and public health. At the individual vehicle level, the impacts of cheating, tampering, or malfunctioning emission control systems can be substantial. The removal or deactivation of emission control systems increases emissions—for instance, typically 70% (NOx and EGR), a factor of 3 or more (NOx and SCR), and a factor of 25–100 (PM and DPF). Our analysis shows significant uncertainty and (geographic) variability regarding the occurrence of cheating and tampering by vehicle owners. The available evidence suggests that fleet-wide impacts of cheating and tampering on emissions are undeniable, substantial, and cannot be ignored. The presence of a relatively small fraction of high-emitters, due to either cheating, tampering, or malfunctioning, causes excess pollution that must be tackled by environmental authorities around the world, in particular in emerging economies, where millions of used ICE vehicles from the US and EU end up. Modernized in-service emission programs designed to efficiently identify and fix large faults are needed to ensure that the benefits of modern vehicle technologies are not lost. Effective programs should address malfunctions, engine problems, incorrect repairs, a lack of servicing and maintenance, poorly retrofitted fuel and emission control systems, the use of improper or low-quality fuels and tampering. Periodic Test and Repair (PTR) is a common in-service program. We estimate that PTR generally reduces emissions by 11% (8–14%), 11% (7–15%), and 4% (−1–10%) for carbon monoxide (CO), hydrocarbons (HC), and oxides of nitrogen (NOx), respectively. This is based on the grand mean effect and the associated 95% confidence interval. PTR effectiveness could be significantly higher, but we find that it critically depends on various design factors, including (1) comprehensive fleet coverage, (2) a suitable test procedure, (3) compliance and enforcement, (4) proper technician training, (5) quality control and quality assurance, (6) periodic program evaluation, and (7) minimization of waivers and exemptions. Now that both particulate matter (PM, i.e., DPF) and NOx (i.e., SCR) emission controls are common in all modern new diesel vehicles, and commonly the focus of cheating and tampering, robust measurement approaches for assessing in-use emissions performance are urgently needed to modernize PTR programs. To increase (cost) effectiveness, a modern approach could include screening methods, such as remote sensing and plume chasing. We conclude this study with recommendations and suggestions for future improvements and research, listing a range of potential solutions for the issues identified in new and in-service vehicles. Full article
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14 pages, 1234 KiB  
Article
Exploring the Potential of Plant Cytokinins Against Common Human Pathogens: In Vitro Assessment and In Silico Insights
by Jelena Lazarević, Aleksandar Veselinović, Marija Stojiljković, Miloš Petrović, Pierangela Ciuffreda and Enzo Santaniello
Plants 2025, 14(12), 1749; https://doi.org/10.3390/plants14121749 - 7 Jun 2025
Viewed by 629
Abstract
Cytokinins, plant hormones derived from adenine, are best known for regulating growth and stress responses in plants. Recent findings suggest they may also influence microbial viability, yet their direct antimicrobial potential remains underexplored. This study evaluates the antimicrobial activities of four natural cytokinins [...] Read more.
Cytokinins, plant hormones derived from adenine, are best known for regulating growth and stress responses in plants. Recent findings suggest they may also influence microbial viability, yet their direct antimicrobial potential remains underexplored. This study evaluates the antimicrobial activities of four natural cytokinins (iPA, B, K, and p-T) and their N9-ribosides (iPAR, BR, KR, and p-TR) against selected human pathogens. Using the broth microdilution method, we assessed their effects on Gram-positive and Gram-negative bacteria, as well as fungal strains. While Gram-negative species showed no susceptibility, all tested compounds exhibited bacteriostatic activity against Bacillus subtilis and Enterococcus faecalis. Most notably, kinetin (K) and kinetin riboside (KR) displayed strong antifungal activity against Candida albicans, with MIC values comparable to the reference drug nystatin. Molecular docking studies supported these findings by showing that K and KR form favorable interactions with two validated antifungal targets in Candida albicans: secreted aspartic proteinase 3 (SAP3) and dihydrofolate reductase (DHFR). This is, to our knowledge, the first report linking natural cytokinins to direct antifungal action against C. albicans supported by in silico evidence. These findings highlight the potential of K and KR as promising leads for the development of cytokinin-based antifungal agents. Full article
(This article belongs to the Special Issue Plant-Derived Natural Products: Development and Utilization)
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14 pages, 3094 KiB  
Article
Evaluation of the Antileishmanial Activity of Some Benzimidazole Derivatives Using In Vitro and In Silico Techniques
by Mustafa Eser, İbrahim Çavuş, Aybüke Züleyha Kaya, Asaf Evrim Evren and Leyla Yurttaş
Vet. Sci. 2025, 12(6), 550; https://doi.org/10.3390/vetsci12060550 - 5 Jun 2025
Viewed by 542
Abstract
Benzimidazole derivatives are well known for their anthelmintic activity. Investigating the potential efficacy of new derivatives of this class against various parasites is essential to identify novel drug candidates. For this purpose, an in-house molecular database was screened, and four benzimidazole-based molecules were [...] Read more.
Benzimidazole derivatives are well known for their anthelmintic activity. Investigating the potential efficacy of new derivatives of this class against various parasites is essential to identify novel drug candidates. For this purpose, an in-house molecular database was screened, and four benzimidazole-based molecules were chosen to evaluate antiprotozoal activity. The compounds (K1K4) had been previously synthesized through a four-step procedure. The potential in vitro cytotoxic properties of the compounds were assessed against the Leishmania (L.) major strain and L929 mouse fibroblast cells. The tests indicated that K1 (3-Cl phenyl) demonstrated an antileishmanial effect (IC50 = 0.6787 µg/mL) and cytotoxicity at elevated concentrations (CC50 = 250 µg/mL) in healthy cells. These findings were comparable to those of AmpB. The antileishmanial activity values were determined as follows: K2; 8.89 µg/mL, K3; 45.11 µg/mL, K4; and 69.19 µg/mL. The CC50 values were determined as follows: K2, 63 µg/mL; K3; 0.56 µg/mL; and K4, 292 µg/mL. Molecular docking and dynamic simulations were conducted to elucidate the potential mechanisms of action of the test substances. In silico investigations indicated interactions between the compounds and the active site of pteridine reductase 1 (PTR1), which is a biosynthetic enzyme essential for parasite proliferation. N-alkyl benzimidazole-based compounds exhibit potential inhibitory activity against L. (L.) major promastigotes. Therefore, these findings suggest that in vivo evaluation is warranted, and structural modifications may lead to the identification of more effective antileishmanial agents. Full article
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25 pages, 6243 KiB  
Article
A Passive Time Reversal Method with a Metamodel for Underwater Source Localization
by Jiang Liu and Sheng Li
J. Mar. Sci. Eng. 2025, 13(6), 1082; https://doi.org/10.3390/jmse13061082 - 29 May 2025
Viewed by 316
Abstract
A passive time reversal method with a metamodel (PTR-MM) is proposed to improve underwater source localization under ocean conditions. PTR-MM eliminates model mismatch errors by replacing the conventional sound propagation model with a Kriging metamodel. This metamodel is optimally constructed based on measured [...] Read more.
A passive time reversal method with a metamodel (PTR-MM) is proposed to improve underwater source localization under ocean conditions. PTR-MM eliminates model mismatch errors by replacing the conventional sound propagation model with a Kriging metamodel. This metamodel is optimally constructed based on measured sound field data. The method combines a metamodel with a passive time reversal (PTR) process to generate a focused sound field whose intensity peaks correspond to source positions. In numerical simulations using the KRAKEN model in a range-independent waveguide, PTR-MM accurately localizes single and multiple sources, is insensitive to mismatches in key environmental parameters, and maintains unbiased performance down to −20 dB signal-to-noise ratios (SNRs). Experimental validation on the SWellEx-96 Event S5 dataset confirms that PTR-MM outperforms conventional PTR in both single- and dual-source localizations, achieving most mean absolute percentage errors (MAPEs) below 10% when trained and tested in consistent environments. Further studies reveal that localization accuracy depends primarily on signal quality, array aperture, and element spacing, rather than on source frequency. However, PTR-MM performance degrades if the metamodel is trained in an environment that differs from the test conditions. The above findings demonstrate the potential of combining PTR with a metamodel for robust and real-time localization. Full article
(This article belongs to the Section Ocean Engineering)
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19 pages, 3126 KiB  
Article
Characterization and Expression Analysis of PUB Gene Family Involved in Drought Stress Response in Trifoliate Orange (Poncirus trifoliata)
by Bobo Song, Sanpeng Jin, Xuchen Gong, Yong Liu, Dechun Liu, Li Yang, Wei Hu, Liuqing Kuang and Jie Song
Horticulturae 2025, 11(6), 604; https://doi.org/10.3390/horticulturae11060604 - 29 May 2025
Viewed by 414
Abstract
The U-box E3 ubiquitin ligase (PUB) gene family plays an important role in regulating plant responses to abiotic stress. Poncirus trifoliata (trifoliate orange), a citrus rootstock with notable cold, drought, and salt tolerance, serves as an excellent model for studying stress-responsive genes. In [...] Read more.
The U-box E3 ubiquitin ligase (PUB) gene family plays an important role in regulating plant responses to abiotic stress. Poncirus trifoliata (trifoliate orange), a citrus rootstock with notable cold, drought, and salt tolerance, serves as an excellent model for studying stress-responsive genes. In this study, a total of 47 PUB genes (PtrPUBs) were identified in the trifoliate orange genome. Chromosomal distribution analysis indicated that PtrPUB genes were unevenly distributed across nine trifoliate orange chromosomes. Phylogenetic tree analysis indicated that 170 PUB proteins from trifoliate orange, Arabidopsis thaliana, and tomato were clustered into five subfamilies. Gene structure, conserved domain, and motif analyses revealed diverse exon–intron and motif organizations of PtrPUB genes, suggesting potential functional differentiation among PtrPUBs. Cis-acting analysis indicated that the promoters of PtrPUB genes harbor elements related to hormone signaling (ABA, MeJA), drought stress, and low-temperature responses. Transcriptomic data and qRT-PCR results suggested that PtrPUB genes are responsive to ABA and dehydration treatments. This study provides a foundation for understanding the functional roles of PUB genes in trifoliate orange and offers insights for improving stress tolerance in citrus breeding programs. Full article
(This article belongs to the Special Issue New Insights into Breeding and Genetic Improvement of Fruit Crops)
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19 pages, 4558 KiB  
Article
Immunogenicity Evaluation of Epitope-Based Vaccine on Target of RNAIII-Activating Protein (TRAP) of Staphylococcus Aureus
by Simiao Yu, Di Yao, Xintong Wang, Wei Yu, Yuhua Wei, Wei Liu, Liquan Yu, Jinzhu Ma, Chunyu Tong, Jing Chen, Yongzhong Yu, Baifen Song and Yudong Cui
Biology 2025, 14(6), 616; https://doi.org/10.3390/biology14060616 - 27 May 2025
Viewed by 628
Abstract
Staphylococcus aureus is a leading cause of severe infections in humans and animals, and the emergence of multidrug-resistant strains highlights the need to develop effective vaccines to prevent such diseases. Epitope-based vaccines use short antigen-derived peptides corresponding to immune epitopes, which are administered [...] Read more.
Staphylococcus aureus is a leading cause of severe infections in humans and animals, and the emergence of multidrug-resistant strains highlights the need to develop effective vaccines to prevent such diseases. Epitope-based vaccines use short antigen-derived peptides corresponding to immune epitopes, which are administered to trigger protective humoral and cellular immune responses. In this study, in silico MHC affinity measurement methods were used to predict possible binding regions, and five 20-mer synthetic TRAP peptides (TRAPP) were synthesized. Epitope-based vaccines, named PT and PTR, incorporating the identified CD4+ T and B cell epitopes, were constructed. Peptides TRAP20–39 and TRAP94–113 elicited significant peptide-stimulated T-cell proliferation responses in vivo. Additionally, high levels of IFN-γ and IL-17A, along with moderate levels of IL-4, were detected in ex vivo stimulated CD4+ T cells isolated from rTRAP- and TRAPP-immunized mice, suggesting that these peptides are classified as Th1 and Th17 epitopes. Immunization with PT or PTR induces robust humoral and cellular immune responses. Moreover, the epitope-based vaccine, PT, exhibited a stronger protective immune response than the intact TRAP in a murine systemic S. aureus infection model. Based on the results presented herein, an epitope-based vaccine is a promising and potentially more effective candidate. Full article
(This article belongs to the Section Infection Biology)
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18 pages, 3712 KiB  
Article
PtrSAUR32 Interacts with PtrPP2C.Ds to Regulate Root Growth in Citrus
by Xiaoli Wang, Xiaoya Li, Saihang Zheng, Fusheng Wang, Shiping Zhu and Xiaochun Zhao
Plants 2025, 14(11), 1579; https://doi.org/10.3390/plants14111579 - 22 May 2025
Viewed by 472
Abstract
Small auxin-up-regulated RNA (SAUR) genes are involved in the regulation of dynamic and adaptive growth in higher plants. However, their function and mode of action in citrus root growth are still unknown. Here, we demonstrate that in Poncirus trifoliata, PtrSAUR32 [...] Read more.
Small auxin-up-regulated RNA (SAUR) genes are involved in the regulation of dynamic and adaptive growth in higher plants. However, their function and mode of action in citrus root growth are still unknown. Here, we demonstrate that in Poncirus trifoliata, PtrSAUR32 acted downstream of the auxin response factor PtrARF8 to regulate root growth by interacting with PtrPP2C.Ds, subfamily type 2C protein phosphatases which interacted with H-ATPase and PtrHA. In this study, several members of SAUR family in Poncirus trifoliata are identified to be associated with the growth and development of the roots. Among them, PtrSAUR32 was found to be highly expressed in the RT (root tip), and the level of its expression was significantly positively corelated to the length of primary roots (p < 0.01). The overexpression of PtrSAUR32 in citrus significantly promoted the growth of primary roots. In PtrSAUR32 transgenic citrus plants, the expressions of several auxin biosynthesis and transport genes were altered in accordance with the expression of PtrSAUR32. Y1H and dual-luciferase reporter assays proved that the expression of PtrSUAR32 is regulated by PtrARF8. Y2H and BiFC assay results indicated that PtrSAUR32 interacted with PtrPP2C.Ds subfamily members PtrPP2C.D1, PtrPP2C.D6, and PtrPP2C.D7, of which PtrPP2C.D7 could interact with PtrHA in vivo. Full article
(This article belongs to the Section Plant Physiology and Metabolism)
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16 pages, 3836 KiB  
Article
Energy Harvesting Microelectromechanical System for Condition Monitoring Based on Piezoelectric Transducer Ring
by Kaixuan Wang, Hao Long, Di Song and Hasan Shariar
Micromachines 2025, 16(6), 602; https://doi.org/10.3390/mi16060602 - 22 May 2025
Cited by 1 | Viewed by 2324
Abstract
For complex mechanical transmission equipment, shaft bearings are usually enclosed together with the shaft in the internal space of the housing to maintain good sealing and reliability. However, it is difficult to monitor the status of the shaft bearing through external sensors on [...] Read more.
For complex mechanical transmission equipment, shaft bearings are usually enclosed together with the shaft in the internal space of the housing to maintain good sealing and reliability. However, it is difficult to monitor the status of the shaft bearing through external sensors on the housing, while internal sensors face challenges in energy supply and data transmission. Therefore, a piezoelectric transducer ring-based energy harvesting microelectromechanical system (PTR-EH-MEMS) is proposed for the condition monitoring of shaft bearings. Specifically, the piezoelectric transducer ring is designed to convert mechanical vibrations into electrical energy, which simultaneously acts as a self-powered monitoring sensor through energy harvesting. In addition, the MEMS is embedded for piezoelectric data processing and condition monitoring of the shaft bearings. To verify the proposed PTR-EH-MEMS, an experimental investigation is implemented under different conditions. The experimental results demonstrate that the system can achieve the maximum DC output of 0.8 V and the root mean square power of 43.979 μW within 128 s, which can effectively identify early-stage bearing faults frequency through a self-powered mode. By combining energy harvesting with condition monitoring capability, the PTR-EH-MEMS offers a compact and sustainable approach for predictive maintenance in rotating machinery, reducing the reliance on external power sources and enhancing the reliability of industrial systems. Full article
(This article belongs to the Special Issue Exploration and Application of Piezoelectric Smart Structures)
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17 pages, 1505 KiB  
Article
De Novo Terpenes Emitted from Juvenile Leaves of Eucalyptus globulus Labill. subsp. globulus
by Anthony J. Winters, Charles H. Hocart, Jörg-Peter Schnitzler, Ina Zimmer, Mark A. Adams, Heinz Rennenberg, Jürgen Kreuzwieser and Claudia Keitel
Molecules 2025, 30(10), 2234; https://doi.org/10.3390/molecules30102234 - 21 May 2025
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Abstract
The contributions of de novo synthesis to terpene emissions from Eucalyptus globulus subsp. globulus were determined by fumigating branchlets with 13CO2 in a gas exchange system. Of more than thirty-four terpenes emitted by this species, only four, i.e., isoprene, iso-valeraldehyde, cis [...] Read more.
The contributions of de novo synthesis to terpene emissions from Eucalyptus globulus subsp. globulus were determined by fumigating branchlets with 13CO2 in a gas exchange system. Of more than thirty-four terpenes emitted by this species, only four, i.e., isoprene, iso-valeraldehyde, cis-ocimene, and trans-caryophyllene, incorporated 13C into the terpene carbon skeleton during the ~5–6 h experiment. 13C incorporation into isoprene and iso-valeraldehyde reached a maximum of ca. 82% of the carbon skeleton, similar to cis-ocimene, with a maximum of 77% 13C incorporation after ~2.5 h exposure to 13CO2. Only ca. 20% of carbon was labelled in trans-caryophyllene after 5–6 h. the incorporation of 13C was observed only in compounds emitted from leaves, and was not detected in either individual oil glands or in bulk leaf tissue. The results suggest the de novo synthesis of some terpenes (isoprene, cis-ocimene, trans-caryophyllene, and iso-valeraldehyde) and their emission is independent of emissions of terpenes stored in oil glands. Full article
(This article belongs to the Section Chemical Biology)
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