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18 pages, 2128 KB  
Article
Effects of Dietary Squalene Supplementation on the Growth Performance and Disease Resistance of Largemouth Bass
by Shan Liu, Mengmeng Chen, Yan Meng, Mingyang Xue, Yong Zhou, Liping Zhang, Peng Chen, Yuding Fan, Yazhen Yang and Zhenyu Huang
Vet. Sci. 2026, 13(5), 448; https://doi.org/10.3390/vetsci13050448 - 1 May 2026
Abstract
Feed supplements play a crucial role in improving and maintaining fish health in modern aquaculture practices. Squalene is a functional lipid naturally present in fatty tissues, possessing numerous beneficial biological properties and wide applications in the food and pharmaceutical industries. In this study, [...] Read more.
Feed supplements play a crucial role in improving and maintaining fish health in modern aquaculture practices. Squalene is a functional lipid naturally present in fatty tissues, possessing numerous beneficial biological properties and wide applications in the food and pharmaceutical industries. In this study, the effects of 100 mg/kg (S1), 200 mg/kg (S2), 300 mg/kg (S3), and 400 mg/kg (S4) of dietary squalene supplementation over four weeks on growth performance, antioxidation, hepatoprotection, hypoxia tolerance, immune relative genes expression, and disease resistance of largemouth bass (Micropterus salmoides) were assessed. The results showed that squalene supplementation significantly increased the weight gain rate (WGR) and specific growth rate (SGR) of largemouth bass (p < 0.05). Serum glucose (GLU) levels were significantly decreased in all squalene-supplemented groups (p < 0.05). Squalene supplementation had minimal effect on serum triglyceride (TG) and total cholesterol in (TCHO) levels. A decrease in malondialdehyde (MDA) level, but accompanied by increases in superoxide dismutase (SOD) and hepatic catalase (CAT) activities, was observed in the S1 group supplemented with squalene. These suggest that squalene may mitigate free radical damage and promote health in largemouth bass. Dietary squalene supplementation enhanced intestinal enzyme activities (trypsin, lipase, and α-amylase) in largemouth bass without inducing any apparent hepatic or histopathological alterations. Squalene supplementation improved hypoxia tolerance and antiviral gene expression (mx, ifn-γ, and irf3) while suppressing the expression of inflammatory cytokine (il-1β, il-8, and tnf-α). The survival rate following LMBRaV infection was significantly higher in the S1 group (100 mg/kg group) compared to the control (p < 0.05). In conclusion, this study demonstrated that adding squalene into the diet of largemouth bass at an optimal level of 100 mg/kg effectively promotes growth performance, enhances digestive enzyme activity and hypoxia tolerance, and modulates lipid metabolism and immune gene expression, thereby contributing to improved resistance against LMBRaV. These findings confirm that squalene can serve as a beneficial functional feed additive in aquaculture. Full article
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16 pages, 11288 KB  
Article
Phillygenin, a Plant-Derived Lignan, Attenuates Renal Inflammation, Fibrosis, and Pyroptosis in a Unilateral Ureteral Obstruction Model
by Yu-Syuan Chen, Shun-Fa Yang, Huey-Liang Kuo, Haw-Ling Chuang, Chang-Mu Chen, Ssu-Chia Lin, Pei-Yu Weng, Chun-Fa Huang, Siao-Syun Guan, Shing-Hwa Liu and Cheng-Tien Wu
Nutrients 2026, 18(9), 1421; https://doi.org/10.3390/nu18091421 - 30 Apr 2026
Abstract
Background/Objectives: Phillygenin (PHI), a natural lignan derived from Forsythia suspensa, has garnered attention for its potential to alleviate chronic diseases, including chronic colitis, pulmonary fibrosis, and diabetes. Chronic kidney disease (CKD) poses a global health challenge, characterized by high morbidity and mortality [...] Read more.
Background/Objectives: Phillygenin (PHI), a natural lignan derived from Forsythia suspensa, has garnered attention for its potential to alleviate chronic diseases, including chronic colitis, pulmonary fibrosis, and diabetes. Chronic kidney disease (CKD) poses a global health challenge, characterized by high morbidity and mortality rates and associated with a spectrum of secondary complications. In this study, we aim to investigate the therapeutic effectiveness of PHI on CKD and also identify molecular signals by using a unilateral ureteral obstruction (UUO) mouse model and in vitro experiments. Methods: C57BL/6 mice were administered PHI at 50 mg/kg/day to assess its therapeutic effectiveness. In vitro, lipopolysaccharide (LPS) and adenosine triphosphate (ATP) were used to induce pyroptosis, also known as pyroptosis, in renal proximal tubular cells (NRK52E). Results: After PHI treatment for 14 consecutive days, the collagen deposition and extracellular matrix (ECM) accumulation, the expression of oxidative stress response proteins (catalase, superoxide dismutase 2, NADPH oxidase 4, and thioredoxin reductase 1), pro-inflammatory markers (TNF-α and Cyclooxygenase-2(COX-2), and infiltration of neutrophils and macrophages were significantly ameliorated in the UUO mice. Interestingly, the pyroptosis-related proteins (NLRP3/Caspase-1/GSDMD/IL-1β) and cell apoptotic death were also conspicuously relieved after treatment with PHI. Furthermore, PHI administration significantly attenuated the ATP/LPS-induced NF-κB/NLRP3/Caspase-1/GSDMD pyroptosis signal pathway in NRK52E cells. Conclusions: These results demonstrate, for the first time, that PHI treatment ameliorates inflammation and the related pyroptosis via inhibitory regulation of the NF-κB/NLRP3/Caspase-1/GSDMD axis, leading to attenuated renal fibrosis and progressive CKD in UUO mice and in vitro. Our findings suggest that PHI could be a nutraceutical candidate for attenuating CKD progression. Full article
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19 pages, 889 KB  
Article
Evidence of Endocrine Disruption and Oxidative Stress in Mytilus galloprovincialis Exposed to 17α-Ethinylestradiol
by Sandra Copeto, Inês João Ferreira, Catarina Mansilha, Armindo Melo, Carla Motta, Marco Silva and Mário Diniz
J. Mar. Sci. Eng. 2026, 14(9), 829; https://doi.org/10.3390/jmse14090829 - 30 Apr 2026
Abstract
The presence of endocrine-disrupting compounds (EDCs) in aquatic environments has raised significant concerns, particularly regarding their impact on marine biota. Among these compounds, 17α-ethinylestradiol (EE2), a synthetic estrogen widely used in oral contraceptives, is highly persistent and biologically active at very low concentrations. [...] Read more.
The presence of endocrine-disrupting compounds (EDCs) in aquatic environments has raised significant concerns, particularly regarding their impact on marine biota. Among these compounds, 17α-ethinylestradiol (EE2), a synthetic estrogen widely used in oral contraceptives, is highly persistent and biologically active at very low concentrations. This study evaluated the effects of EE2 exposure on oxidative stress responses and endocrine disruption in Mytilus galloprovincialis, exposed for 28 days to three EE2 concentrations (10, 30, and 300 ng·L−1). Biomarkers of oxidative stress, including the enzymatic activities of superoxide dismutase (SOD), catalase (CAT), and glutathione-S-transferase (GST), as well as Lipid Peroxidation (MDA levels), total ubiquitin (UBI) and the endocrine disruption marker, vitellogenin-like protein (VTG) were assessed. Results showed significant increase in GST and a decrease in CAT activities followed by an elevation at 300 ng·L−1, slightly higher than control values. Overall, results suggest an enhanced oxidative challenge. No significant changes were detected in MDA and UBI levels. VTG-like protein levels increased according to the EE2 concentrations tested, suggesting an effect on the mussel’s endocrine system. These results show the activation of detoxification and antioxidant defense mechanisms in exposed mussels as a response to mitigate oxidative stress damage. Furthermore, it highlights the importance of using biomarkers in pollution monitoring studies and environmental risk assessment. Full article
(This article belongs to the Special Issue Marine Bivalves Toxicology)
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20 pages, 9594 KB  
Article
Multi-Omics Analysis Identifies SlLhcb13 as a Key Regulator of Tomato Resistance to Botrytis cinerea
by Dan Luo, Xiaojie Peng, Weiqiang Yan, Yujin Wang, Ke Liu, Lixia Li, Zhe Wu, Hongmei Nie, Sheng Sun, Wenhui Sun and Jun Cai
Plants 2026, 15(9), 1360; https://doi.org/10.3390/plants15091360 - 29 Apr 2026
Viewed by 35
Abstract
Gray mold caused by Botrytis cinerea poses a severe threat to tomato production. In this study, physiological, biochemical, transcriptomic, and proteomic analyses were integrated to characterize the dynamic responses of tomato ‘Ailsa Craig’ to B. cinerea infection. During B. cinerea infection, peroxidase (POD) [...] Read more.
Gray mold caused by Botrytis cinerea poses a severe threat to tomato production. In this study, physiological, biochemical, transcriptomic, and proteomic analyses were integrated to characterize the dynamic responses of tomato ‘Ailsa Craig’ to B. cinerea infection. During B. cinerea infection, peroxidase (POD) activity showed a progressive increase, while catalase (CAT) activity was significantly upregulated at 24 hpi and remained stable through 48 hpi. Malondialdehyde (MDA) and hydrogen peroxide (H2O2) contents showed a delayed response, increasing significantly only at 48 hpi, whereas SOD activity exhibited a biphasic pattern. Transcriptome and proteome profiling identified 5824 differentially expressed genes and 124 differentially expressed proteins. Functional enrichment analysis highlighted defense-related pathways, including plant–pathogen interaction, flavonoid biosynthesis, and inositol phosphate metabolism. Notably, the chlorophyll a/b-binding protein SlLhcb13 exhibited post-transcriptional upregulation despite transcriptional suppression. Functional validation demonstrated that overexpression of SlLhcb13 enhanced resistance, whereas silencing increased susceptibility. These findings identify SlLhcb13 as a positive regulator linking photosynthesis to immunity and provide new insights into the defense mechanisms of tomato. Full article
(This article belongs to the Topic Genetic Breeding and Biotechnology of Garden Plants)
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13 pages, 1382 KB  
Article
Integrated Assessment of Metal-Related Toxicity in a Sentinel Marine Plant, Posidonia oceanica, Under Realistic Multi-Element Exposure
by Paolo Cocci, Martina Fattobene, Raffaele Emanuele Russo, Mario Berrettoni and Francesco Alessandro Palermo
Int. J. Mol. Sci. 2026, 27(9), 3946; https://doi.org/10.3390/ijms27093946 - 29 Apr 2026
Viewed by 133
Abstract
Mediterranean meadows of Posidonia oceanica are chronically exposed to complex mixtures of environmental contaminants, including metals and trace elements derived from coastal urbanization, maritime traffic, and industrial activities. This study aimed to assess metal-related toxicity in P. oceanica by integrating multi-element burden analysis [...] Read more.
Mediterranean meadows of Posidonia oceanica are chronically exposed to complex mixtures of environmental contaminants, including metals and trace elements derived from coastal urbanization, maritime traffic, and industrial activities. This study aimed to assess metal-related toxicity in P. oceanica by integrating multi-element burden analysis with a panel of oxidative stress biomarkers. Concentrations of a wide suite of elements were quantified in samples of internal (juvenile), intermediate, and external (adult) leaves, reflecting the ontogenetic structure of the plant. Oxidative responses were evaluated using five biomarkers [i.e., hydrogen peroxide (H2O2), lipid peroxidation (TBARS), superoxide dismutase (SOD), glutathione S-transferase (GST), and catalase (CAT)] measured on each leaf compartment. Biomarker data were standardized and integrated into a merged Stress Index summarizing overall physiological toxicity. Associations between individual elements, the sum of all measured elements (ΣallElements), the Stress Index, and single biomarkers were explored using Pearson correlation analysis. Juvenile leaves exhibited the highest Stress Index values, elevated H2O2 and TBARS, and marked activation of SOD and GST, indicating early oxidative toxicity. Intermediate leaves showed a trend toward increased CAT activity, not reaching statistical significance, along with minimal damage, suggesting effective detoxification, whereas adult leaves accumulated higher levels of Fe, Ni, and Pb, but displayed moderate stress responses. Overall, leaf-class structure strongly modulated both exposure and toxicological response. The integration of ΣAllElements with multi-biomarker indices provides a robust framework for diagnosing metal-related toxicity in P. oceanica under realistic multi-element exposure scenarios. Full article
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11 pages, 1921 KB  
Article
An Exploratory Investigation into the Effects of Methanol and Ethylene Glycol on the Growth and Development of Medaka (Oryzias melastigma) and Yellowstripe Goby (Mugilogobius chulae)
by Zhenxiong Yang, Minxia Zhang, Tongfei Xu, Shasha Qi, Lu Tang, Juan Guo, Chuqian Lu and Shaobo Ma
Toxics 2026, 14(5), 380; https://doi.org/10.3390/toxics14050380 - 29 Apr 2026
Viewed by 173
Abstract
This study aimed to investigate the acute and chronic toxic effects of two thermodynamic inhibitors (methanol and ethylene glycol) widely used in deep-sea oil and gas operations on two typical marine fish species, the medaka (Oryzias melastigma) and the yellowstripe goby [...] Read more.
This study aimed to investigate the acute and chronic toxic effects of two thermodynamic inhibitors (methanol and ethylene glycol) widely used in deep-sea oil and gas operations on two typical marine fish species, the medaka (Oryzias melastigma) and the yellowstripe goby (Mugilogobius chulae), to assess their potential ecological risks in marine environments. The 96-h median lethal concentration (LC50) was determined through acute toxicity tests. A 56-day chronic toxicity test was conducted to evaluate the effects on fish growth (body length) and the antioxidant defense system, specifically the activities of superoxide dismutase (SOD) and catalase (CAT). The results revealed marked species-specific differences. In terms of acute toxicity, medaka exhibited exceptionally high sensitivity to ethylene glycol (LC50 15.77 g/L), while the yellowstripe goby showed greater tolerance (LC50 22.17 g/L). Chronic exposure led to concentration-dependent growth inhibition in both species, and medaka showed significantly higher mortality than yellowstripe goby. Under methanol exposure, medaka exhibited significantly higher mortality (30–45%) than yellowstripe goby (5–20%). When exposed to ethylene glycol, medaka showed markedly high mortality (55–85%), while yellowstripe goby mortality remained below 15%. At the molecular level, both chemicals induced oxidative stress, but the response patterns of the antioxidant enzymes (SOD and CAT) were species-specific, indicating differences in toxic mechanisms and detoxification capacities. Methanol and ethylene glycol pose non-negligible ecotoxicological risks to marine fish, and the toxicity intensity is influenced by species specificity, exposure concentration, and the effectiveness of the antioxidant defense system. This study emphasizes that environmental risk assessments for such chemicals must fully account for species differences and sublethal effects, providing critical scientific evidence for formulating precise environmental safety standards for marine hydrocarbon exploitation. Full article
(This article belongs to the Special Issue Fish Physiological Responses to Environmental Stressors)
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27 pages, 5355 KB  
Article
Species-Specific Effects of Humic Substances and Mycorrhiza on Antioxidant Defense and Metal Stress Tolerance in Cannabis sativa, Sorghum sudanense × bicolor, and Miscanthus × giganteus Under Field Conditions
by Karolina Jaros-Tsoj, Artur Nowak, Jolanta Jaroszuk-Ściseł, Piotr Sugier, Danuta Sugier, Francois Rineau, Jaco Vangronsveld and Małgorzata Wójcik
Int. J. Mol. Sci. 2026, 27(9), 3942; https://doi.org/10.3390/ijms27093942 - 28 Apr 2026
Viewed by 220
Abstract
Abiotic stresses, including heavy metal contamination, can severely impair plant growth and antioxidative defense. However, their adverse effects may be mitigated through sustainable strategies such as biostimulant application. This study investigated the effects of humic substances (HSs), alone or combined with mycorrhizal inoculation [...] Read more.
Abiotic stresses, including heavy metal contamination, can severely impair plant growth and antioxidative defense. However, their adverse effects may be mitigated through sustainable strategies such as biostimulant application. This study investigated the effects of humic substances (HSs), alone or combined with mycorrhizal inoculation (M), on oxidative stress and antioxidative responses in Cannabis sativa, Sorghum sudanense × bicolor, and Miscanthus × giganteus grown under field conditions on metal-contaminated agricultural soil exceeding regulatory thresholds for Zn, Pb, and Cd. Plant growth, lipid peroxidation, stress-related metabolites (proline, sugars), antioxidative enzyme activities (catalase, CAT; ascorbate peroxidase, APX; guaiacol peroxidase, GOPX; glutathione reductase, GR, and superoxide dismutase, SOD), and leaf metal concentrations were analyzed. Biostimulants increased proline and sugars in Sorghum (by up to 55% and 80%, respectively), accompanied by reduced oxidative stress indicators and improved biomass (by 26%). In Cannabis, higher Cd and Pb concentrations following biostimulant treatments were associated with increased SOD, APX, and GR activities (by 33–267%), without affecting growth. In Miscanthus, increased lipid peroxidation (by 37–60%) occurred alongside enhanced GR and APX activities. These results indicate strong species-specific responses and absence of consistent synergistic effects of HSs and M, highlighting distinct physiological strategies of stress adaptation and antioxidative defense on metal-contaminated soils. Future research should address physiological and molecular mechanisms underlying these responses. Full article
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22 pages, 2151 KB  
Review
Nanozyme-Driven Signal Amplification in Cancer Biosensing: Innovations Toward Precision and Point-of-Care Oncology
by Victor Akpe and Ian E. Cock
Micromachines 2026, 17(5), 541; https://doi.org/10.3390/mi17050541 - 28 Apr 2026
Viewed by 199
Abstract
This review evaluates recent progress in nanozyme-based biosensors for detecting circulating tumour cells, nucleic acids, and protein biomarkers, with particular attention to how peroxidase-, oxidase-, and catalase-like reactions enhance signal generation across electrochemical, optical, and microfluidic platforms. The roles of iron oxide–gold composites, [...] Read more.
This review evaluates recent progress in nanozyme-based biosensors for detecting circulating tumour cells, nucleic acids, and protein biomarkers, with particular attention to how peroxidase-, oxidase-, and catalase-like reactions enhance signal generation across electrochemical, optical, and microfluidic platforms. The roles of iron oxide–gold composites, silica nanostructures, quantum dots, and hybrid nanomaterials in improving analytical performance, enabling multiplexed detection, and facilitating assay miniaturization are critically assessed. Advances such as amplification-free detection approaches, smartphone-compatible point-of-care systems, and AI-assisted data analysis are discussed in relation to their translational potential. Key barriers, including regulatory requirements, reproducibility concerns, and manufacturing scalability, are also evaluated. By integrating mechanistic understanding with practical considerations for clinical deployment, this review outlines how next-generation nanozyme-based biosensors may strengthen early cancer detection, real-time monitoring, and precision oncology. Full article
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18 pages, 318 KB  
Article
In Ovo Administration of Silver Nanoparticles Enhances Post-Hatch Growth Performance, Metabolic and Physiological Responses, and Immune Function in Broiler Chickens
by Hanan Al-Khalaifah, Samar A. Tolba, Inas I. Ismail, Azza A. Megahid, Eman S. Osman, Ahmed H. Rabie and Ahmed Gouda
Animals 2026, 16(9), 1349; https://doi.org/10.3390/ani16091349 - 28 Apr 2026
Viewed by 178
Abstract
This study examined the effects of in ovo silver nanoparticle (AgNP) administration on broiler chicken hatchability, growth performance, physiological and metabolic status, antioxidant capacity, and immune responses. A total of 300 fertilized Cobb500 eggs were assigned to five treatments (5 replicates/treatment, 12 eggs/replicate): [...] Read more.
This study examined the effects of in ovo silver nanoparticle (AgNP) administration on broiler chicken hatchability, growth performance, physiological and metabolic status, antioxidant capacity, and immune responses. A total of 300 fertilized Cobb500 eggs were assigned to five treatments (5 replicates/treatment, 12 eggs/replicate): non-injected control; vehicle control (0.2 mL sterile 0.9% NaCl/egg); and AgNPs at 10, 15, or 20 ppm injected into their amniotic sac on day (d) 18 of incubation. After hatching, 48 chicks/treatment were assigned to 6 replicate cages (8 birds/replicate) for 35 d. In ovo AgNP administration improved hatchability and reduced embryonic mortality (p < 0.05). Post-hatch, birds originating from AgNP-injected eggs had an improved body weight, body weight gain, and feed conversion ratio, with lower feed intake (p < 0.05). AgNP administration elevated hemoglobin, packed cell volume, growth hormone, serum proteins, alanine aminotransferase activity, and creatinine levels (p < 0.05). AgNPs also improved the lipid profile and increased total antioxidant capacity, catalase and superoxide dismutase activities, lysozyme activity, complement 3, and cytokine concentrations and reduced malondialdehyde levels (p < 0.05). Effects were dose-dependent, with 15 ppm generally leading to better outcomes. These findings suggest that 15 ppm AgNP in ovo administration enhances embryonic viability, physiological resilience, and post-hatch performance in broiler chickens. Full article
(This article belongs to the Special Issue Metabolic, Health, and Productivity Challenges in Poultry Production)
21 pages, 1172 KB  
Article
Effects of Dietary Arachidonic Acid Concentration on Growth, Fatty Acid Profile, and Inflammatory/Redox Status of Juvenile Clam Sinonovacula constricta
by Yuxiang Zhu, Yueyue Fu, Kai Liao, Yang Liu, Yang Zhang and Jilin Xu
Fishes 2026, 11(5), 262; https://doi.org/10.3390/fishes11050262 - 27 Apr 2026
Viewed by 114
Abstract
Dietary arachidonic acid (ARA) is essential for aquatic animal growth and health, but studies in bivalves are still limited. Here, microcapsule diets with increasing ARA concentrations (ARA1-6 groups: 0.35, 3.01, 5.25, 6.88, 8.69, and 10.27 mg g−1 dry matter) were prepared by [...] Read more.
Dietary arachidonic acid (ARA) is essential for aquatic animal growth and health, but studies in bivalves are still limited. Here, microcapsule diets with increasing ARA concentrations (ARA1-6 groups: 0.35, 3.01, 5.25, 6.88, 8.69, and 10.27 mg g−1 dry matter) were prepared by spray drying, and clam Sinonovacula constricta juveniles were fed these diets for 14 days. Results showed that dietary ARA concentrations did not significantly affect clams’ survival, weight gain, and shell length gain rates. The clams in the ARA6 group had significantly higher crude lipid content than those in the other microcapsule groups. The ARA concentrations in the clams increased with higher dietary ARA, while n-3 polyunsaturated fatty acids (PUFAs) and eicosapentaenoic acid (EPA) concentrations decreased. The mRNA levels of cyclooxygenase 2 and 5-lipoxygenase type 2 were significantly higher in the ARA5 and ARA6 groups compared to the ARA1 group. The mRNA levels of 5-lipoxygenase type 3, toll-like receptor 4, and nuclear factor-kappa b p50 (nfκb p50) were significantly higher in the ARA6 group compared to the ARA1 group. As dietary ARA concentrations increased, the mRNA levels of glutamate–cysteine ligase catalytic subunit and glutathione S-transferase, along with malondialdehyde (MDA) content, increased in the clams. Additionally, the superoxide dismutase and catalase activities in the ARA5 and ARA6 groups were significantly higher than those in the ARA1 and ARA2 groups. Clam ARA content, acting as a central node, showed very strong positive correlations with MDA and cyclooxygenase 2, and very strong negative correlations with EPA and the n-3/n-6 PUFA ratio. Our results revealed that high dietary ARA, while not affecting growth, reduced the n-3/n-6 PUFA ratio and induced a response characterized by the upregulation of NF-κB and Nrf2 pathway genes in S. constricta. Full article
(This article belongs to the Section Aquatic Invertebrates)
16 pages, 1122 KB  
Article
Molecular Epidemiology, Hematobiochemical Alterations, and Oxidative Stress-Induced Genotoxicity of Equine Trypanosomiasis in Pakistan
by Waqas Ahmad, Naeem Rasool, Qurat ul Ain, Usama Bin Naeem, Muhammad Azeem, Umbreen Anwar, Tehreem Fayyaz, Zeba Amjad, Muhammad Yasin Tipu and Mehmood Ahmad
Microbiol. Res. 2026, 17(5), 86; https://doi.org/10.3390/microbiolres17050086 - 27 Apr 2026
Viewed by 114
Abstract
Trypanosoma evansi (T. evansi) infection poses a significant health threat to equines. This study was aimed to assess the prevalence, risk factors, hematobiochemical alterations, and oxidative stress-mediated genotoxicity associated with equine trypanosomiasis in the Rahim Yar Khan District. This cross-sectional study [...] Read more.
Trypanosoma evansi (T. evansi) infection poses a significant health threat to equines. This study was aimed to assess the prevalence, risk factors, hematobiochemical alterations, and oxidative stress-mediated genotoxicity associated with equine trypanosomiasis in the Rahim Yar Khan District. This cross-sectional study was conducted on 384 equines from October 2024 to September 2025. Blood samples were collected for thin blood film microscopy and PCR assay using RoTat 1.2 primers. Hematological indices were analyzed with an automated hematology analyzer; serum biochemical parameters were quantified via standard assays. Oxidative stress markers, including malondialdehyde (MDA), catalase (CAT), superoxide dismutase (SOD), and reduced glutathione (GSH), were also measured. Genotoxicity was evaluated using the alkaline comet assay. Statistical analyses included the chi-square test, logistic regression, and independent t-tests. T. evansi was detected in 5.99% of samples by microscopy and 10.16% by PCR, with no significant association with species, age, or sex. Infected equines exhibited significant reductions in hemoglobin (5.4 ± 0.6 vs. 10.8 ± 0.5 g/dL; p < 0.0001), total serum protein (2.1 ± 0.3 vs. 5.8 ± 0.2 g/dL; p < 0.0001), albumin, and globulin, alongside elevated hepatic enzymes, blood urea nitrogen, and creatinine (all p < 0.01). Oxidative stress was confirmed by increased MDA (p < 0.0001) and decreased CAT activity (p < 0.001). Genotoxicity was significantly higher in infected animals (genetic damage index; 1.12 ± 0.08 vs. 0.40 ± 0.01; p < 0.01). This study provides the first integrated assessment of molecular epidemiology and oxidative stress-mediated genotoxicity in equines in this region, suggesting the pathogenic impact of the infection and targeted diagnostics for disease management strategies. Full article
15 pages, 683 KB  
Article
Investigating the Effects of Nanogels in Promoting Protein Crystallization
by Lata Govada, Beijia Wang, Yanmin Li, Emmanuel Saridakis and Naomi E. Chayen
Int. J. Mol. Sci. 2026, 27(9), 3879; https://doi.org/10.3390/ijms27093879 - 27 Apr 2026
Viewed by 97
Abstract
X-ray crystallography is still the most widely used and versatile method for structural studies of biological macromolecules. This study concerns the application of nanogels to facilitate protein crystallization, a prerequisite for X-ray crystallography. Nanogels (NGs) are nano-sized, highly crosslinked polymeric particles that have [...] Read more.
X-ray crystallography is still the most widely used and versatile method for structural studies of biological macromolecules. This study concerns the application of nanogels to facilitate protein crystallization, a prerequisite for X-ray crystallography. Nanogels (NGs) are nano-sized, highly crosslinked polymeric particles that have been extensively studied for chemical catalysis and drug delivery but not for protein crystal nucleation. The efficacy of six types of nanogels (three N-isopropylacrylamide-based and three acrylamide-based) was tested, with promising results. They were subsequently functionalised with active hydroxyl groups for further testing. Both functionalised and non-functionalised nanogels were tested on model (trypsin, thaumatin, proteinase K, ferritin and catalase) and target proteins (glulisine, α-crustacyanin and acriflavine resistance protein subunit AcrB) using both manual and automated techniques. All nanogels were found to be effective in promoting protein crystallization in both screening and optimization trials, giving crystal ‘hits’ that would have otherwise been missed. Overall, the functionalised nanogels were more effective. Nanogel effects are proposed to be due to a combination of surface porosity and surface chemistry. Full article
19 pages, 2642 KB  
Article
Novel NSAID Analogs Exhibit Anti-Leukemic Activity Through Modulation of Apoptotic and Survival Pathways
by Hind A. Alkhatabi, Mohammed Basabrain, Alaa G. Alahmadi, Shiekhah M. Alzahrani, Yosra A. Muhammad, Maha Almuhaiyawi, Maha M. Alreemi, Reem M. Alotibi, Roaa M. Alreemi, Heba A. Alkhattabi, Reem N. Hassan, Wedad M. Al-Bishri, Mohammed El-Mezgueldi and Abdelsattar M. Omar
Int. J. Mol. Sci. 2026, 27(9), 3850; https://doi.org/10.3390/ijms27093850 - 26 Apr 2026
Viewed by 269
Abstract
Acute myeloid leukemia (AML) is a complex blood cancer that primarily affects relapsing or refractory patients receiving conventional chemotherapy. Nonsteroidal anti-inflammatory drugs (NSAIDs) have anticancer properties with restricted clinical efficacy attributable to cyclooxygenase (COX)-induced toxicities. To address this issue, a group of benzylamide [...] Read more.
Acute myeloid leukemia (AML) is a complex blood cancer that primarily affects relapsing or refractory patients receiving conventional chemotherapy. Nonsteroidal anti-inflammatory drugs (NSAIDs) have anticancer properties with restricted clinical efficacy attributable to cyclooxygenase (COX)-induced toxicities. To address this issue, a group of benzylamide analogs of the classical NSAIDs (NSI-1–NSI-9) were developed and synthesized to mask the carboxylic acid moiety and minimize COX-induced adverse effects while maintaining anticancer activity. The cytotoxic effect of such substances has been demonstrated in some leukemia cell lines (HL-60, MV4-11, KG1a, and K562). NSI-5 exerted the highest anti-leukemic activity among these sulindac analogs, as determined at a sub-micromolar level in all cell lines studied, by IC50. This mechanistic data also demonstrated that NSI-5 induced apoptosis that was dose-dependent, especially in HL-60 cell lines, and increased the sub-G1 cell fraction. This apoptotic process was also accompanied by a significant decrease in mitochondrial membrane potential, which is characteristic of the induction of the intrinsic apoptotic process. Interestingly, NSI-5 decreased the intracellular reactive oxygen species (ROS) and the expression of most antioxidants (catalase and glutathione synthetase), as well as the redox balance. Gene characterization in vitro also suggested activation of apoptotic pathways, where expression of Bax, Bak1, and Caspase-3 increased, suggesting a potential p53-independent apoptotic pathway, in contrast to control for Bcl-2 expression. Collectively, these findings indicate that NSI-5 is a promising in vitro anti-leukemic lead compound, with activity associated with mitochondrial dysfunction and altered redox regulation. The observed effects are consistent with previously reported COX-independent activity of structurally related NSAID derivatives, and support further investigation of NSI-5 in preclinical models. Full article
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17 pages, 4283 KB  
Article
Celastrol Ameliorates Renal Injury in Spontaneously Hypertensive Rats by Activating the Nrf2/Ho-1 Signaling Pathway to Alleviate Oxidative Stress
by Yijie Deng, Jichun Wang, Xiping Liu, Xiuwen Wang, Hua Li, Bo Gu, Min Zhang, Renjun Wang and Yi Yang
Int. J. Mol. Sci. 2026, 27(9), 3849; https://doi.org/10.3390/ijms27093849 - 26 Apr 2026
Viewed by 231
Abstract
Celastrol (CSL), a natural triterpenoid extracted from Tripterygium wilfordii, demonstrates a wide range of biological activities. In this study, we explored whether CSL alleviates kidney damage in spontaneously hypertensive rats (SHRs) through the modulation of the Nrf2/Ho-1 pathway, a crucial target in [...] Read more.
Celastrol (CSL), a natural triterpenoid extracted from Tripterygium wilfordii, demonstrates a wide range of biological activities. In this study, we explored whether CSL alleviates kidney damage in spontaneously hypertensive rats (SHRs) through the modulation of the Nrf2/Ho-1 pathway, a crucial target in renal injury models. A total of 40 male SHRs, aged 6–8 weeks, were randomly allocated to four groups: the control group (CON, serving as the healthy control), the spontaneously hypertensive rat group (SHR), the SHR group treated with low-dose CSL (L-CSL + SHR, 0.5 mg/kg/d), and the SHR group treated with high-dose CSL (H-CSL + SHR, 1 mg/kg/d). All drugs were formulated using physiological saline as the solvent and administered via intraperitoneal injection. The control group received an equivalent volume of physiological saline via intraperitoneal injection, and all groups underwent continuous daily administration for 6 weeks. The results indicated that, in comparison with the control group, the serum levels of angiotensin, angiotensin-converting enzyme, and aldosterone in the SHR group were relatively high, and CSL treatment further downregulated these indices. Simultaneously, CSL downregulated pro-inflammatory factors (tumor necrosis factor-α and interleukin-1β) and upregulated interleukin-6. Regarding renal function-related indicators, CSL reduced malondialdehyde levels and enhanced the activities of antioxidant enzymes, such as superoxide dismutase, glutathione peroxidase, and catalase. Moreover, CSL inhibited the overexpression of Keap1. Significantly, the mRNA levels of Nrf2, Nqo1, and Ho-1 in the CSL-treated groups were notably higher than those in the SHR group. These findings suggest that CSL mitigates renal pathological damage in SHR by activating the Nrf2/Ho-1 pathway, offering a potential therapeutic approach for hypertension-induced renal injury. Full article
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Article
Effect of Short-Term Static Magnetic Field Pretreatment on Cold-Storage Quality and Phenolic Metabolism of Blueberries
by Ying Lu, Hui Liu, Zhenzhen Lv, Chengheng Li, Muhammad Nawaz, Qiang Zhang, Wenbo Yang, Jiechao Liu, Wenqiang Guan and Zhonggao Jiao
Foods 2026, 15(9), 1505; https://doi.org/10.3390/foods15091505 - 25 Apr 2026
Viewed by 199
Abstract
Blueberries are widely consumed due to their richness in nutrients, yet they are also prone to quality deterioration after being harvested, even at cold temperatures. Non-thermal physical technology is an important auxiliary method worth considering for maintaining the quality of this fruit while [...] Read more.
Blueberries are widely consumed due to their richness in nutrients, yet they are also prone to quality deterioration after being harvested, even at cold temperatures. Non-thermal physical technology is an important auxiliary method worth considering for maintaining the quality of this fruit while refrigerated. In this study, a static magnetic field (SMF) was applied as a complementary pretreatment strategy prior to cold storage of blueberries. The optimal SMF parameters were identified as 5 mT exposure for 12 h, as this significantly retarded decay and softening. The contents of ascorbic acid, total polyphenols, flavonoids and proanthocyanidins were elevated by 20.0%, 17.7%, 23.9%, and 9.1%, respectively. Concurrently, DPPH (1,1-diphenyl-2-picrylhydrazyl) radical-scavenging capacity, catalase (CAT), and superoxide dismutase (SOD) activity markedly improved. Targeted metabolomic analysis revealed that SMF pretreatment significantly regulated polyphenol metabolic pathways and redirected polyphenol biosynthesis toward more stable and functional compounds, including three hydroxycinnamic acids, quercetin, dihydromyricetin, glycosylated hesperetin, and acylated delphinidin derivates. The synergistic effect of these SMF-elevated phenolics and the reinforced antioxidant system preserved the overall cold-storage quality of blueberries. These findings underscore the potential of SMF pretreatment as an effective physical technique for reducing postharvest blueberry losses. Full article
(This article belongs to the Special Issue Postharvest Storage and Preservation Technologies for Agri-Food)
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