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21 pages, 9119 KB  
Article
Multi-Omics Integration Reveals Key Genes, Metabolites and Pathways Underlying Meat Quality and Intramuscular Fat Deposition Differences Between Tibetan Pigs and Duroc × Tibetan Crossbred Pigs
by Junda Wu, Qiuyan Huang, Baohong Li, Zixiao Qu, Xinming Li, Fei Li, Haiyun Xin, Jie Wu, Chuanhuo Hu, Sen Lin, Xiangxing Zhu, Dongsheng Tang, Chuang Meng, Zongliang Du, Erwei Zuo, Fanming Meng and Sutian Wang
Animals 2026, 16(2), 214; https://doi.org/10.3390/ani16020214 - 11 Jan 2026
Viewed by 51
Abstract
The sensory quality of pork constitutes a complex phenotype that arises from the interplay between genetic factors and environmental conditions. As a local pig breed in China, Tibetan pigs (TPs) are known for their high-quality meat. However, their slow growth rate and low [...] Read more.
The sensory quality of pork constitutes a complex phenotype that arises from the interplay between genetic factors and environmental conditions. As a local pig breed in China, Tibetan pigs (TPs) are known for their high-quality meat. However, their slow growth rate and low production efficiency limit their large-scale breeding. We have used Duroc as a hybrid sire to improve TP. Our study found that TPs have higher intramuscular fat content and higher levels of monounsaturated fatty acids. Duroc × Tibetan crossbred pigs (DZs) not only retain the paternal high productivity but also inherit the superior meat quality of the maternal parent. Transcriptome analysis identified IL6, GPX1, GPX3, AOX1, ALDH7A1, PTGS2, NFKBIA, ADIPOQ and PPARG as being involved in affecting meat quality. Metabolomic analysis found that betaine, carnosine, L-carnitine, and lysophosphatidylcholine were important components that affect meat quality. Joint analysis further reveals that the expression of ATF4, DGKB, GNMT, and ADSL genes is closely related to arachidonic acid, lysophosphatidylcholine, betaines, and hypoxanthine, ultimately affecting the quality of the meat. By comprehensively analyzing the carcass and meat qual Full article
(This article belongs to the Special Issue Livestock Omics)
16 pages, 1019 KB  
Article
Investigation of the Impact of Wastewater from Waste Oil In-Stallation on the Activated Sludge Process, to Ensure the Proper Operation of Municipal Wastewater Treatment Plant
by Agnieszka Bluszcz, Krzysztof Barbusiński, Barbara Pieczykolan and Mohamed Alwaeli
Water 2026, 18(1), 108; https://doi.org/10.3390/w18010108 - 1 Jan 2026
Viewed by 309
Abstract
The study evaluated the feasibility of using the activated sludge process to treat real wastewater from used oil installations containing petroleum hydrocarbons, boron (B), and adsorbable organic halides (AOX). The aim was to determine the maximum ratio of this wastewater that could be [...] Read more.
The study evaluated the feasibility of using the activated sludge process to treat real wastewater from used oil installations containing petroleum hydrocarbons, boron (B), and adsorbable organic halides (AOX). The aim was to determine the maximum ratio of this wastewater that could be added to the influent without impairing treatment efficiency. Tested shares ranged from 0.50% to 1.90%. An initial 1.30% of the tested share caused process instability, reflected in the elevated total nitrogen (TN) levels in treated wastewater. After reducing the share to 0.50%, an adaptation of the activated sludge was observed, manifested by a decrease in TN concentration to below 15.0 mg N/L. For the most favorable share of 1.60% (0.38 ± 0.10 kgBOD5/kgMLSS d, 0.51 ± 0.14 kgCOD/kgMLSS d), the removal efficiencies of chemical oxygen demand (COD), biochemical oxygen demand (BOD5), TN, and total phosphorus (TP) were 95.0% ± 1.5, 99.1% ± 0.2, 89.3% ± 2.7, and 94.0% ± 5.0, respectively. Increasing the share to 1.90% decreased treatment efficiency and exceedances of COD, BOD5, TN, and TP occurred. At this ratio, an increase in ammonium nitrogen (NH4+-N) and TN concentrations was observed, indicating the inhibition of nitrification. However, the average concentrations of mineral oil index, AOX and B in the treated wastewater remained within permissible levels throughout the study. Full article
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19 pages, 3727 KB  
Article
Extracellular Phosphate Availability Impacts Aspergillus terreus Itaconic Acid Fermentation via Biomass-Specific Product Yield
by Ákos P. Molnár, István Bakondi-Kovács, Vivien Bíró, Alexandra Márton, István S. Kolláth, Erzsébet Fekete, Norbert Ág, Erzsébet Sándor, András Csótó, Béla Kovács, Christian P. Kubicek and Levente Karaffa
J. Fungi 2026, 12(1), 14; https://doi.org/10.3390/jof12010014 - 25 Dec 2025
Viewed by 361
Abstract
Itaconic acid (IA) is an important bio-based platform chemical produced via submerged fermentation by the filamentous Ascomycete Aspergillus terreus. In this study, we examined the impact of initial phosphate concentration on IA production from D-glucose and D-xylose in optimized, manganese-limited fermentations. Nine [...] Read more.
Itaconic acid (IA) is an important bio-based platform chemical produced via submerged fermentation by the filamentous Ascomycete Aspergillus terreus. In this study, we examined the impact of initial phosphate concentration on IA production from D-glucose and D-xylose in optimized, manganese-limited fermentations. Nine phosphate concentrations ranging from 0.04 to 4 g L−1 were tested, and representative low (0.04 g L−1), optimal (0.1 g L−1), and high (0.8 g L−1) conditions were analyzed in detail in controlled, 6 L scale bioreactors. Phosphate availability primarily influenced biomass formation and the biomass-to-product ratio rather than directly affecting IA accumulation. Both lower- and higher-than-optimal phosphate concentrations decreased the volumetric and specific IA yields, while the highest productivity was observed at 0.1 g L−1. Expression of the aoxA gene, encoding the cyanide-resistant alternative oxidase (AOX), and AOX enzymatic activity were inversely correlated with extracellular phosphate concentration, consistent with a role in redox homeostasis under phosphate-limited conditions. In contrast, total respiration rates and pellet-type morphology remained unaffected. These findings indicate that phosphate acts mainly as a secondary modulator of IA fermentation performance through its influence on biomass formation, whereas other metabolic constraints play a more dominant role in controlling IA overflow in A. terreus. Full article
(This article belongs to the Special Issue Biotechnological Applications of Fungi)
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31 pages, 1574 KB  
Review
Nanoparticle-Based Assays for Antioxidant Capacity Determination
by Jolanta Flieger, Natalia Żuk, Ewelina Grabias-Blicharz, Piotr Puźniak and Wojciech Flieger
Antioxidants 2025, 14(12), 1506; https://doi.org/10.3390/antiox14121506 - 15 Dec 2025
Viewed by 579
Abstract
Thanks to both endogenous and exogenous antioxidants (AOs), the antioxidant defense system ensures redox homeostasis, which is crucial for protecting the body from oxidative stress and maintaining overall health. The food industry also exploits the antioxidant properties to prevent or delay the oxidation [...] Read more.
Thanks to both endogenous and exogenous antioxidants (AOs), the antioxidant defense system ensures redox homeostasis, which is crucial for protecting the body from oxidative stress and maintaining overall health. The food industry also exploits the antioxidant properties to prevent or delay the oxidation of other molecules during processing and storage. There are many classical methods for assessing antioxidant capacity/activity, which are based on mechanisms such as hydrogen atom transfer (HAT), single electron transfer (SET), electron transfer with proton conjugation (HAT/SET mixed mode assays) or the chelation of selected transition metal ions (e.g., Fe2+ or Cu1+). The antioxidant capacity (AOxC) index value can be expressed in terms of standard AOs (e.g., Trolox or ascorbic acid) equivalents, enabling different products to be compared. However, there is currently no standardized method for measuring AOxC. Nanoparticle sensors offer a new approach to assessing antioxidant status and can be used to analyze environmental samples, plant extracts, foodstuffs, dietary supplements and clinical samples. This review summarizes the available information on nanoparticle sensors as tools for assessing antioxidant status. Particular attention has been paid to nanoparticles (with a size of less than 100 nm), including silver (AgNPs), gold (AuNPs), cerium oxide (CeONPs) and other metal oxide nanoparticles, as well as nanozymes. Nanozymes belong to an advanced class of nanomaterials that mimic natural enzymes due to their catalytic properties and constitute a novel signal transduction strategy in colorimetric and absorption sensors based on the localized surface plasmon resonance (LSPR) band. Other potential AOxC sensors include quantum dots (QDs, <10 nm), which are particularly useful for the sensitive detection of specific antioxidants (e.g., GSH, AA and baicalein) and can achieve very good limits of detection (LOD). QDs and metallic nanoparticles (MNPs) operate on different principles to evaluate AOxC. MNPs rely on optical changes resulting from LSPR, which are monitored as changes in color or absorbance during synthesis, growth or aggregation. QDs, on the other hand, primarily utilize changes in fluorescence. This review aims to demonstrate that, thanks to its simplicity, speed, small sample volumes and relatively inexpensive instrumentation, nanoparticle-based AOxC assessment is a useful alternative to classical approaches and can be tailored to the desired aim and analytes. Full article
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18 pages, 527 KB  
Article
Mexican Fruits of the Stenocereus Genus: Characterization, Physicochemical, Nutritional, and Content of Bioactive Compounds
by Alma Delia Noriega-Juárez, Yolanda Nolasco-González, Oswaldo Alejandro Arellano-Machuca, Diego Rafael Montoya-Carvajal, Brandon Alexis López-Romero, Javier Alberto Navarro-Mendoza and Efigenia Montalvo-González
Compounds 2025, 5(4), 55; https://doi.org/10.3390/compounds5040055 - 2 Dec 2025
Viewed by 358
Abstract
Stenocereus fruits are appreciated for their flavor and color, and their cultivation is highly sustainable, as they grow in arid zones without the need for fertilizers or agrochemicals. However, their nutritional and bioactive composition remains underexplored. This study evaluated the physicochemical and nutritional [...] Read more.
Stenocereus fruits are appreciated for their flavor and color, and their cultivation is highly sustainable, as they grow in arid zones without the need for fertilizers or agrochemicals. However, their nutritional and bioactive composition remains underexplored. This study evaluated the physicochemical and nutritional properties, bioactive compound content, and antioxidant capacity (AOX) of Stenocereus thurberi (red, white, purple, and orange), Stenocereus martinezzi (red), and Stenocereus gummosus (red). All fruits exhibited low total soluble solids (12.6 ± 0.2–14.7 ± 0.3 °Brix), acidity (0.81 ± 0.03–1.12 ± 0.03%), and moderate dietary fiber content (3.71 ± 0.05–4.86 ± 0.09%). S. martinezzi stood out for its high levels of betalains (33.7 ± 0.65 mg/100 g_fw), vitamin E (84.7 ± 0.2 µg/100 g_fw), and vitamin C (147.6 ± 11.4 mg/100 g_fw). At the same time, potassium, magnesium, and calcium were the predominant minerals in all samples. S. gummosus showed the highest total soluble phenols (120.6 ± 2.2 mg/100 g_fw) and was also notable for its flavonoid content. Flavonoids, hydroxycinnamic, and hydroxybenzoic acids were detected in all fruits. Red fruits had the highest AOX levels, followed by white, orange, and purple varieties. In conclusion, these fruits are nutritious, low in sugars, and rich in bioactive compounds, suggesting their potential as functional foods, particularly beneficial for individuals with chronic degenerative diseases. Full article
(This article belongs to the Special Issue Phenolic Compounds: Extraction, Chemical Profiles, and Bioactivity)
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16 pages, 745 KB  
Article
The Effect of Fermentation with Saccharomyces cerevisiae on the Release of Bound Phenolic Compounds from Wheat Bran and Its Effect on Antioxidant Capacity
by Víctor Manuel Ocaño-Higuera, Guadalupe López-Avilés, Francisco Javier Almendariz-Tapia, Carmen Lizette Del-Toro-Sánchez, José Agustín Tapia-Hernández, Alba Mery Garzón-García, Octavio Dublán-García, Ariel Alain Vergel-Alfonso, Gustavo Adolfo González-Aguilar, Santiago Valdez-Hurtado, Hebert Jair Barrales-Cureño, José Rogelio Ramos-Enríquez and Dalila Fernanda Canizales-Rodríguez
Processes 2025, 13(11), 3506; https://doi.org/10.3390/pr13113506 - 31 Oct 2025
Viewed by 641
Abstract
Wheat bran (WB) is a rich source of phenolic compounds (PCs) with antioxidant capacity (AOX). Approximately 90% of these PCs are bound to the cell wall matrix, which limits their bioavailability. Fermentation with Saccharomyces cerevisiae is an effective strategy to release these bound [...] Read more.
Wheat bran (WB) is a rich source of phenolic compounds (PCs) with antioxidant capacity (AOX). Approximately 90% of these PCs are bound to the cell wall matrix, which limits their bioavailability. Fermentation with Saccharomyces cerevisiae is an effective strategy to release these bound phenolics. This study aimed to evaluate the effect of fermentation on the release of bound PCs in WB and their AOX during an in vitro digestion system. WB was fermented with Saccharomyces cerevisiae for 2, 4, and 6 days and subsequently subjected to simulated digestion. Free PCs were extracted with methanol, while bound PCs were obtained through alkaline hydrolysis. Total PCs were quantified using the Folin–Ciocalteu method, and AOX was assessed through DPPH, TEAC, and FRAP assays. The content of bound PCs significantly increased after fermentation (p < 0.05): 30.24 ± 0.06 mg GAE/g (day 2), 27.18 ± 0.40 mg GAE/g (day 4), and 28.41 ± 0.40 mg GAE/g (day 6), compared with unfermented WB (7.7 ± 0.21 mg GAE/g) (p < 0.05). AOX was notably enhanced; DPPH reached its peak on day 4 (47.38 ± 0.07 µmol TE/g) (p < 0.05). TEAC was highest on day 2 (26.20 ± 0.43 µmol TE/g) compared with the control (20.14 ± 0.22 µmol TE/g) (p < 0.05) and FRAP showed a slight improvement on day 6 (57.38 ± 0.10 µmol TE/g) relative to the control (56.22 ± 0.13 µmol TE/g) (p < 0.05). Fermentation with Saccharomyces cerevisiae promotes the release of bound PCs in WB and enhances its AOX, highlighting its potential as a functional food ingredient. Full article
(This article belongs to the Special Issue Green Chemistry: From Wastes to Value-Added Products (2nd Edition))
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14 pages, 2092 KB  
Article
Toxicity of Tris(2-chloroethyl) Phosphate (TCEP) to Alfalfa’s Root System: An Insight into TCEP’s Damage to Morphology, Respiration, and Antioxidant Systems
by Meijun Liu, Liangzhu Gong, An Yan, Wenjing Liu, Haojie Li and Peiyi Guo
Agronomy 2025, 15(11), 2483; https://doi.org/10.3390/agronomy15112483 - 25 Oct 2025
Viewed by 736
Abstract
Tris(2-chloroethyl) phosphate (TCEP), as an organophosphate contaminant, poses a significant threat to the growth and development of plants, especially roots. This study aimed to clarify the mechanisms of TCEP’s toxicity and damage to root systems, as well as the mechanisms of its damage [...] Read more.
Tris(2-chloroethyl) phosphate (TCEP), as an organophosphate contaminant, poses a significant threat to the growth and development of plants, especially roots. This study aimed to clarify the mechanisms of TCEP’s toxicity and damage to root systems, as well as the mechanisms of its damage to the respiration and energy metabolism of alfalfa root cells. The results showed that TCEP obviously affected the root length, root surface area, root volume, and root diameter of alfalfa. With increasing stress intensity, the total mitochondrial respiration rate and Cytochrome C Oxidase (COX) pathway respiration rate progressively declined, while the Alternative Oxidase (AOX) pathway respiration rate and its proportion of total respiration gradually rose. In addition, adenosine triphosphate (ATP) content and root vigor were significantly reduced. Moreover, with an increase in TCEP concentration, root superoxide anion radical content in alfalfa root cells was significantly elevated, while superoxide dismutase (SOD) and catalase (CAT) activities were significantly lowered, and ascorbate peroxidase (APX) and peroxidase (POD) activities were significantly enhanced. The present study indicated that respiration was disrupted, causing a lack of ATP in root cells under TCEP. Both the overproduction of reactive oxygen species (ROS) from the mitochondrial respiratory electron transport chain (mECT) and the deficiency of ROS-scavenging enzymes caused ROS accumulation, which led to the destruction of the cell membrane structure and exacerbated the disruption of the respiratory metabolism. The disruption of the conversion and reuse of energy by TCEP affected root growth and development. Full article
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17 pages, 1293 KB  
Article
A Diamine Oxidase from Glutamicibacter halophytocola for the Degradation of Histamine and Tyramine in Foods
by Lucas Kettner, Alexander Freund, Anna Bechtel, Judit Costa-Catala and Lutz Fischer
Foods 2025, 14(17), 3093; https://doi.org/10.3390/foods14173093 - 3 Sep 2025
Cited by 1 | Viewed by 2030
Abstract
A novel diamine oxidase (DAO) was discovered in the bacterium Glutamicibacter halophytocola (DAO-GH). The gene of DAO-GH was integrated into the genome of the yeast Komagataella phaffii and recombinantly produced under control of the methanol-inducible AOX1 promoter in a bioreactor cultivation. A high [...] Read more.
A novel diamine oxidase (DAO) was discovered in the bacterium Glutamicibacter halophytocola (DAO-GH). The gene of DAO-GH was integrated into the genome of the yeast Komagataella phaffii and recombinantly produced under control of the methanol-inducible AOX1 promoter in a bioreactor cultivation. A high DAO activity of 70.2 ± 5.2 µkat/Lculture (5.25 ± 0.22 µkat/gprotein) was yielded after 90 h of cultivation. The DAO-GH was partially purified by the polyethyleneimine precipitation of nucleic acids, fractionated ammonium sulfate precipitation and hydrophobic interaction chromatography, resulting in a specific DAO activity of 19.7 µkat/gProtein. The DAO-GH was then biochemically investigated regarding its potential for histamine and tyramine degradation in fermented foods and the human small intestine. Interestingly, the DAO-GH showed activity even at a low pH of 5 and low temperature of 6 °C. Both histamine and tyramine were effectively degraded and DAO-GH showed especially very high affinity towards tyramine (Km of 0.009 mM). The DAO-GH was shown to be capable of degrading around 20% of the initially applied histamine in tuna paste (pH 5.6) at 5 °C within 24 h and completely degraded the histamine in a simulated intestinal fluid within 1.5 h in bioconversion experiments. The DAO-GH was spray-dried for the production of a storable enzyme preparation. Only around 17% of activity were lost in this process and the DAO-GH remained stable at room temperature for at least 3 months. The discovery of this DAO with its very advantageous biochemical properties allows the preparation of histamine-reduced or -free fermented foods by a simple enzymatic treatment or the treatment of histamine intolerance symptoms as a dietary supplement or medicine. Full article
(This article belongs to the Section Food Physics and (Bio)Chemistry)
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13 pages, 2216 KB  
Article
Formation of AOX and Cl/Br/I-THMs During NaClO Disinfection: A Comprehensive Study
by Chao Feng, Tongyu Liu, Yongping Shi, Jimin Shen, Lei Yuan and Yuanqing Guo
Water 2025, 17(17), 2563; https://doi.org/10.3390/w17172563 - 30 Aug 2025
Viewed by 1327
Abstract
The interaction of disinfectants with bromide/iodide ions and natural organic matter (NOM) generates brominated/iodinated disinfection byproducts (Br/I-DBPs), known for their heightened cytotoxicity and genotoxicity relative to chlorinated DBPs. This study investigated Br/I-DBP formation during sodium hypochlorite (NaClO) disinfection of lignin-containing synthetic water by [...] Read more.
The interaction of disinfectants with bromide/iodide ions and natural organic matter (NOM) generates brominated/iodinated disinfection byproducts (Br/I-DBPs), known for their heightened cytotoxicity and genotoxicity relative to chlorinated DBPs. This study investigated Br/I-DBP formation during sodium hypochlorite (NaClO) disinfection of lignin-containing synthetic water by quantifying adsorbable organic halogens (AOX) and trihalomethanes (THMs). Disinfection of bromide-containing water yielded a bromide ion (Br) to adsorbable organic bromine (AOBr) conversion rate of approximately 60%, with bromine within THMs accounting for about 30% of the total AOBr, indicating significant brominated DBP formation where THMs represent a major fraction. Conversely, iodide ion (I) conversion to adsorbable organic iodine (AOI) is minimal, suggesting negligible iodinated DBP formation under NaClO disinfection. Examination of key parameters revealed that brominated THM (Br-THM) formation decreased with increasing lignin concentration, while iodinated THM (I-THM) formation increased. The effect of chlorine dose differed: Br-THM formation exhibited an initial increase followed by a decrease at higher doses, whereas I-THM formation consistently decreased. Both Br-THM and I-THM formation increased with higher bromide or iodide ion concentration and with increasing pH. These results highlight the distinct pathways and influencing factors governing brominated versus iodinated DBP formation in lignin-containing waters disinfected with hypochlorite. Full article
(This article belongs to the Special Issue Novel Advanced Oxidation Technology for Water Treatment)
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16 pages, 4508 KB  
Article
Characterization and Transcriptome Analysis Reveal Abnormal Pollen Germination in Cytoplasmic Male Sterile Tomato
by Kosuke Kuwabara and Tohru Ariizumi
Int. J. Mol. Sci. 2025, 26(17), 8337; https://doi.org/10.3390/ijms26178337 - 28 Aug 2025
Cited by 1 | Viewed by 964
Abstract
Cytoplasmic male sterility (CMS) is a plant trait wherein plants cannot develop normal male organs because of the mitochondrial genes. Although the mitochondrial gene orf137 has been identified as the CMS-causing gene in tomatoes, its function remains unclear. In this study, we characterized [...] Read more.
Cytoplasmic male sterility (CMS) is a plant trait wherein plants cannot develop normal male organs because of the mitochondrial genes. Although the mitochondrial gene orf137 has been identified as the CMS-causing gene in tomatoes, its function remains unclear. In this study, we characterized the sterile male phenotypes and analyzed the CMS pollen transcriptome. Microscopic and calcium imaging analyses revealed that CMS pollen exhibited abnormal germination from multiple apertures, accompanied by elevated calcium concentrations and vesicle accumulation, which are typically observed in pollen tube tips. RNA-Seq analysis revealed 440 differentially expressed genes, including four pectin methylesterase inhibitor (PMEI) genes that were highly expressed in the pollen. PME activity was significantly reduced in CMS pollen, suggesting its association with abnormal pollen germination. ATP and reactive oxygen species (ROS) levels, which are key mediators of mitochondrial retrograde signaling (MRS), remained unchanged in CMS pollen, and the expression of the mitochondrial stress marker AOX1a was not elevated. These findings suggest that orf137 triggers an alternative MRS pathway independent of ATP or ROS, potentially leading to PMEI upregulation and abnormal pollen germination. Our results reveal a previously unrecognized mechanism of CMS-induced male sterility in tomatoes involving nuclear gene regulation through unconventional mitochondrial signaling. Full article
(This article belongs to the Special Issue Solanaceae Biology: From Genetics to Metabolism)
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29 pages, 13368 KB  
Article
Systems Network Integration of Transcriptomic, Proteomic, and Bioinformatic Analyses Reveals the Mechanism of XuanYunNing Tablets in Meniere’s Disease via JAK-STAT Pathway Modulation
by Zhengsen Jin, Chunguo Wang, Yifei Gao, Xiaoyu Tao, Chao Wu, Siyu Guo, Jiaqi Huang, Jiying Zhou, Chuanqi Qiao, Keyan Chai, Hua Chang, Chun Li, Xun Zou and Jiarui Wu
Pharmaceuticals 2025, 18(9), 1266; https://doi.org/10.3390/ph18091266 - 25 Aug 2025
Viewed by 1515
Abstract
Background: Meniere’s disease (MD) is a rare inner ear disorder characterized by endolymphatic hydrops and symptoms such as vertigo and hearing loss, with no curative treatment currently available. XuanYunNing tablets (XYN) have been clinically used to treat MD, but their molecular mechanisms remain [...] Read more.
Background: Meniere’s disease (MD) is a rare inner ear disorder characterized by endolymphatic hydrops and symptoms such as vertigo and hearing loss, with no curative treatment currently available. XuanYunNing tablets (XYN) have been clinically used to treat MD, but their molecular mechanisms remain unclear. Objective: This study aimed to systematically evaluate the pharmacological effects of XYN in a guinea pig model of MD and to elucidate the underlying molecular mechanisms of both MD pathogenesis and XYN intervention through integrated multi-omics analyses, including transcriptomics, proteomics, and bioinformatics. Methods: A guinea pig model of endolymphatic hydrops was induced by intraperitoneal injection of desmopressin acetate (dDAVP). Pharmacodynamic efficacy was evaluated via behavioral scoring and histopathological analysis. The differentially expressed genes (DEGs) and differentially expressed proteins (DEPs) modulated by XYN treatment were identified using high-throughput transcriptomic and proteomic sequencing. These data were integrated through multi-omics bioinformatic analysis. Key molecular targets and signaling pathways were further validated using RT-qPCR and Western blotting. Results: Pharmacological evaluations showed that guinea pigs in the model group exhibited a 26% increase in endolymphatic hydrops area, while high-dose XYN treatment reduced this area by 19% and significantly improved functional parameters, including overall physiological condition (e.g., weight and general appearance), auricular reflexes to low-, medium-, and high-frequency sound stimuli, nystagmus, and the righting reflex. High-throughput sequencing combined with integrative omics analysis identified 513 potential molecular targets of XYN. Subsequent network and module analyses pinpointed the JAK-STAT signaling pathway as the central axis. Mendelian randomization (MR) analysis further supported a causal relationship between MD and metabolic, immune, and inflammatory traits, reinforcing the central role of JAK-STAT signaling in both MD progression and XYN-mediated intervention. Mechanistic studies confirmed that XYN downregulated IFNG, IFNGR1, JAK1, p-STAT3/STAT3, and AOX at both mRNA and protein levels, thereby inhibiting aberrant JAK-STAT pathway activation in MD model animals. In addition, a total of 125 chemical constituents were identified in XYN by UHPLC-MS analysis. ZBTB20 and other molecules were identified as potential blood-based biomarkers for MD. Conclusions: This study reveals that XYN alleviates MD symptoms by disrupting a pathological cycle driven by JAK-STAT signaling, inflammation, and metabolic dysfunction. These findings support the clinical potential of XYN in the treatment of Meniere’s disease and may inform the development of novel therapeutic strategies. Full article
(This article belongs to the Special Issue Network Pharmacology of Natural Products, 2nd Edition)
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16 pages, 3118 KB  
Article
Functional Investigation of Mutant Vespa tropica Hyaluronidase Produced in Pichia pastoris: In Silico Studies and Potential Applications
by Piyapon Janpan, Bernhard Schmelzer, Anuwatchakij Klamrak, Jaran Nabnueangsap, Yutthakan Saengkun, Shaikh Shahinur Rahman, Prapenpuksiri Rungsa, Diethard Mattanovich and Sakda Daduang
Fermentation 2025, 11(9), 497; https://doi.org/10.3390/fermentation11090497 - 25 Aug 2025
Viewed by 1733
Abstract
The hyaluronidase enzyme derived from Vespa tropica (VesT2a) venom contains two putative catalytic residues. Herein, a double mutation was introduced into VesT2a at its catalytic sites by substituting Asp107 and Glu109 with Asn and Gln, respectively, to assess their essential roles in enzymatic [...] Read more.
The hyaluronidase enzyme derived from Vespa tropica (VesT2a) venom contains two putative catalytic residues. Herein, a double mutation was introduced into VesT2a at its catalytic sites by substituting Asp107 and Glu109 with Asn and Gln, respectively, to assess their essential roles in enzymatic function. We used Pichia pastoris to produce the mutated version of the VesT2a (mVesT2a) protein, and the process was more efficient when employing the methanol-inducible promoter (PAOX1) compared to the constitutive promoter (PGAP). In bioreactor scale-up, P. pastoris harboring the pAOX1-αMF-mVesT2a plasmid secreted 34.03 ± 2.31 mg/L of mVesT2a, with an apparent molecular mass of 46.6 kDa, retaining only 2.9% of hyaluronidase activity, thus indicating successful mutation. The newly developed indirect ELISA-based method using mVesT2a demonstrated its potential as an alternative approach for measuring hyaluronic acid (HA) at low concentrations and was also used to confirm HA-binding capacity. In silico docking and molecular dynamics simulations further supported the stable interaction of the mVesT2a–HA complex while suggested other surrounded acidic amino acid residues, which may play a minor role in HA degradation, supporting the remaining activity observed in the in vitro experiments. Full article
(This article belongs to the Section Microbial Metabolism, Physiology & Genetics)
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18 pages, 1441 KB  
Article
Cutinase Production in Komagataella phaffii (Pichia pastoris): Performance Differences Between Host Strains
by Andrea Sabido-Ramos, Montserrat Tagle-Gil, Krystel Estefany León-Montes, José Augusto Castro-Rodríguez and Amelia Farrés
Fermentation 2025, 11(8), 483; https://doi.org/10.3390/fermentation11080483 - 20 Aug 2025
Viewed by 1660
Abstract
The Pichia system has been exploited for decades as a host for recombinant protein production, but there is still an information gap regarding problems that may arise with its use. The application of strains based on the methanol-induced alcohol oxidase 1 (AOX1) promoter [...] Read more.
The Pichia system has been exploited for decades as a host for recombinant protein production, but there is still an information gap regarding problems that may arise with its use. The application of strains based on the methanol-induced alcohol oxidase 1 (AOX1) promoter may represent a safety issue, and its performance varies among strains. In this study, the ability of a Komagataella phaffii MutS KM71H strain to produce recombinant cutinases was evaluated and compared to that of the more widely used Mut+ X-33 strain. The effects of the nature of the cutinase (ANCUT1 and ANCUT3, from Aspergillus nidulans), methanol level, and inoculum concentrations were evaluated in shake flasks containing a complex medium. Higher activities and volumetric cutinase productivity were observed at lower induction cell densities (0.5%) for the MutS KM71H aox1::pPICZα-A-ANCUT1 strain, while a higher one (2%) yielded better results in KM71H aox1::pPICZα-A-ANCUT3. The best inoculum and inducer conditions for both strains yielded similar results. The behavior of the different cutinases in the MutS or Mut+ genetic background was opposed: strain KM71H aox1::pPICZα-A-ANCUT3 produced 19% more activity than strain X-33 aox1::pPICZα-A-ANCUT3, while the ANCUT1 containing strain produced significantly higher activity in the X-33 Mut+ strain. These results indicate that MutS strains are viable host options without the complications of rapidly growing methanol strains. The effect of the gene structure being expressed is a phenomenon that needs further exploration. Full article
(This article belongs to the Section Microbial Metabolism, Physiology & Genetics)
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17 pages, 3691 KB  
Article
Evaluating the Effect of Fresh and Aged Antioxidant Formulations in Skin Protection Against UV Damage
by John Ivarsson, Patricia Brieva, Hina Choudhary and Giuseppe Valacchi
Cosmetics 2025, 12(4), 166; https://doi.org/10.3390/cosmetics12040166 - 7 Aug 2025
Cited by 1 | Viewed by 3277
Abstract
Introduction: Extrinsic skin damage is often a result of oxidative stress caused by exposure to environmental factors such as ultraviolet (UV) radiation, ozone (O3), and various pollutants. As a result, topical antioxidants have been evaluated for their effectiveness in mitigating or [...] Read more.
Introduction: Extrinsic skin damage is often a result of oxidative stress caused by exposure to environmental factors such as ultraviolet (UV) radiation, ozone (O3), and various pollutants. As a result, topical antioxidants have been evaluated for their effectiveness in mitigating or reversing skin damage caused by environmental factors. Topical antioxidants containing a combination of l-ascorbic acid, tocopherol, and ferulic acid have significantly improved markers of skin health after exposure to environment-induced skin damage. However, research suggests that l-ascorbic acid and tocopherol tend to be relatively unstable, possibly affecting their efficacy against outdoor stressor damage. It has been shown that ferulic acid significantly improves the stability of both l-ascorbic acid and tocopherol, but its long-term stabilization effects on these antioxidants are relatively unknown. Material and Methods: This study evaluated the time-dependent effectiveness of a topical antioxidant mix containing 15% l-ascorbic acid, 1% tocopherol, and 0.5% ferulic acid (AOX) on UV-induced skin damage. Skin biopsies (12 mm, n = 60) were placed in a 6-well plate with medium and incubated at 37 °C and 5% CO2 overnight. The day after, skin samples were pretreated with 10 µL of differently aged AOX (0-, 6-, 12-, and 36-month-old) and then exposed to different doses of UV light (100, 200, 400 mJ/cm2) daily over four days. AOX formulations were stored in a cool, dry, and dark place at approximately 20–22 °C during the whole study. This study evaluated 4-hydroxynonenal (4-HNE) and 8-hydroxy-2′-deoxyguanosine (8-OHdG) as oxidative damage and skin DNA damage markers, Collagen1 and Filaggrin as skin structure, and IL-8 and Nrf2 as inflammatory and defensive response. Results: UV exposure significantly increased oxidative and inflammatory markers in human skin explants affecting also filaggrin and collagen levels. However, pre-treatment with the antioxidant formulation, particularly in its younger formulations (0-, 6-, and 12-month-old), significantly reduced the damaging effect of UV. Additionally, all antioxidant formulations effectively mitigated UV-induced damage across all doses. Conclusions: Our results indicate that pre-treatment with this formulation consistently reduces UV-induced oxidative damage and DNA damage in human skin explants, regardless of the formulation age and the discoloration state. Although effective, the protective capacity of aged formulations may be reduced only when extreme UV exposure is tested, a condition that is unlikely to occur under typical environmental conditions. These results support ferulic acid as a stabilization agent for topical antioxidant mixtures. Full article
(This article belongs to the Section Cosmetic Formulations)
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Article
Comparative Quantitative Proteomic Analysis of High and Low Toxin-Producing Karenia brevis Strains Reveals Differences in Polyketide Synthase Abundance and Redox Status of the Proteome
by Kathleen S. Rein, Ricardo Colon, Carlos R. Romagosa, Nicholas R. Ohnikian, Kirstie T. Francis and Samuel R. Rein
Mar. Drugs 2025, 23(7), 291; https://doi.org/10.3390/md23070291 - 17 Jul 2025
Cited by 1 | Viewed by 1566
Abstract
To identify differentially abundant polyketide synthases (PKSs) and to characterize the biochemical consequences of brevetoxin biosynthesis, bottom-up, TMT-based quantitative proteomics and redox proteomics were conducted to compare two strains of the Florida red tide dinoflagellate Karenia brevis, which differ significantly in their [...] Read more.
To identify differentially abundant polyketide synthases (PKSs) and to characterize the biochemical consequences of brevetoxin biosynthesis, bottom-up, TMT-based quantitative proteomics and redox proteomics were conducted to compare two strains of the Florida red tide dinoflagellate Karenia brevis, which differ significantly in their brevetoxin content. Forty-eight PKS enzymes potentially linked to brevetoxin production were identified, with thirty-eight showing up to 16-fold higher abundance in the high-toxin strain. A pronounced shift toward a more oxidized redox state was observed in this strain’s proteome. Notably, 25 antioxidant-related proteins were significantly elevated, including alternative oxidase (AOX), which increased by 17-fold. These results elucidate the cellular consequences of toxin biosynthesis in K. brevis, offer new leads for the study of brevetoxin biosynthesis, and suggest a novel red tide mitigation approach targeting high toxin-producing strains. Full article
(This article belongs to the Section Marine Toxins)
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