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20 pages, 9776 KB  
Article
iTRAQ-Based Proteomics Reveals the Potential Mechanisms Underlying Diet Supplementation with Stevia Isochlorogenic Acid That Alleviates Immunosuppression in Cyclophosphamide-Treated Broilers
by Jiatong Jin, Shuqi Zhao, Pengyu Zhao, Yushuo Zhang, Lifei Wu, Liangfu Zhou, Yasai Sun, Wen Zhao and Qian Zhou
Animals 2026, 16(1), 25; https://doi.org/10.3390/ani16010025 - 22 Dec 2025
Abstract
The extensive use of antibiotics in intensive farming weakens immunity and threatens food safety. Stevia isochlorogenic acid (SICA), a kind of dicaffeoylquinic acid derived from stevia residue, exhibits strong antioxidant activity. This study evaluated the ability of SICA to improve immune function in [...] Read more.
The extensive use of antibiotics in intensive farming weakens immunity and threatens food safety. Stevia isochlorogenic acid (SICA), a kind of dicaffeoylquinic acid derived from stevia residue, exhibits strong antioxidant activity. This study evaluated the ability of SICA to improve immune function in an immunosuppressed broiler model. SICA significantly increased the spleen, thymus, and bursa of Fabricius indices (p < 0.05), alleviated spleen damage, and elevated serum interleukin-2 (IL-2), IL-4, interferon-γ, IL-1β, tumor necrosis factor-α, immunoglobulins (IgA, IgM, IgG), and complement components C3 and C4 (p < 0.05). Isobaric tags for relative and absolute quantification-based proteomics indicated that SICA enhanced splenic immune function by activating cell adhesion molecules, phagosomes, and the intestinal immune network for IgA production pathways. Quantitative PCR analysis showed upregulation of mRNA and protein levels of B-cell receptor, major histocompatibility complex class II, protein tyrosine phosphatase receptor type C, and neutrophil cytosolic factor 2 (p67phox) and downregulation of C-C motif chemokine receptor 9. Molecular docking demonstrated the strongest binding affinity between SICA and p67phox. Overall, SICA effectively alleviated immunosuppression in broiler chickens and represents a promising natural alternative to antibiotic feed additives. Full article
(This article belongs to the Section Poultry)
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34 pages, 3112 KB  
Article
Artificial Intelligence Applied to Soil Compaction Control for the Light Dynamic Penetrometer Method
by Jorge Rojas-Vivanco, José García, Gabriel Villavicencio, Miguel Benz, Antonio Herrera, Pierre Breul, German Varas, Paola Moraga, Jose Gornall and Hernan Pinto
Mathematics 2025, 13(21), 3359; https://doi.org/10.3390/math13213359 - 22 Oct 2025
Viewed by 625
Abstract
Compaction quality control in earthworks and pavements still relies mainly on density-based acceptance referenced to laboratory Proctor tests, which are costly, time-consuming, and spatially sparse. Lightweight dynamic cone penetrometer (LDCP) provides rapid indices, such as qd0 and qd1, [...] Read more.
Compaction quality control in earthworks and pavements still relies mainly on density-based acceptance referenced to laboratory Proctor tests, which are costly, time-consuming, and spatially sparse. Lightweight dynamic cone penetrometer (LDCP) provides rapid indices, such as qd0 and qd1, yet acceptance thresholds commonly depend on ad hoc, site-specific calibrations. This study develops and validates a supervised machine learning framework that estimates qd0, qd1, and Zc directly from readily available soil descriptors (gradation, plasticity/activity, moisture/state variables, and GTR class) using a multi-campaign dataset of n=360 observations. While the framework does not remove the need for the standard soil characterization performed during design (e.g., W, γd,field, and RCSPC), it reduces reliance on additional LDCP calibration campaigns to obtain device-specific reference curves. Models compared under a unified pipeline include regularized linear baselines, support vector regression, Random Forest, XGBoost, and a compact multilayer perceptron (MLP). The evaluation used a fixed 80/20 train–test split with 5-fold cross-validation on the training set and multiple error metrics (R2, RMSE, MAE, and MAPE). Interpretability combined SHAP with permutation importance, 1D partial dependence (PDP), and accumulated local effects (ALE); calibration diagnostics and split-conformal prediction intervals connected the predictions to QA/QC decisions. A naïve GTR-average baseline was added for reference. Computation was lightweight. On the test set, the MLP attained the best accuracy for qd1 (R2=0.794, RMSE =5.866), with XGBoost close behind (R2=0.773, RMSE =6.155). Paired bootstrap contrasts with Holm correction indicated that the MLP–XGBoost difference was not statistically significant. Explanations consistently highlighted density- and moisture-related variables (γd,field, RCSPC, and W) as dominant, with gradation/plasticity contributing second-order adjustments; these attributions are model-based and associational rather than causal. The results support interpretable, computationally efficient surrogates of LDCP indices that can complement density-based acceptance and enable risk-aware QA/QC via conformal prediction intervals. Full article
(This article belongs to the Special Issue Artificial Intelligence and Data Science, 2nd Edition)
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16 pages, 980 KB  
Article
Markers of Antiviral Response in SLE Patients After Vaccination Against SARS-CoV-2
by Michał Komorniczak, Katarzyna Aleksandra Lisowska, Barbara Bułło-Piontecka, Alicja Dębska-Ślizień and Anna Wardowska
Int. J. Mol. Sci. 2025, 26(20), 10241; https://doi.org/10.3390/ijms262010241 - 21 Oct 2025
Viewed by 591
Abstract
Patients with systemic lupus erythematosus (SLE) and lupus nephritis (LN) are at increased risk of severe infections, making effective vaccination strategies essential. While antibody responses to SARS-CoV-2 vaccination have been studied in SLE, less is known about innate immune correlates. Therefore, we evaluated [...] Read more.
Patients with systemic lupus erythematosus (SLE) and lupus nephritis (LN) are at increased risk of severe infections, making effective vaccination strategies essential. While antibody responses to SARS-CoV-2 vaccination have been studied in SLE, less is known about innate immune correlates. Therefore, we evaluated cytokines with a particular emphasis on interferon and chemokine profiles. To fulfill the immunological picture, we also assessed neutralizing antibodies against SARS-CoV-2 variants, lymphocyte subpopulations, and selected gene expression signatures in 33 patients stratified by vaccination status: fully vaccinated (FV, n = 23) and partially vaccinated (PV, n = 10). Serum analyses showed that FV patients exhibited increased type I (IFN-α2, IFN-β) and type III (IFN-λ1, IFN-λ2/3) interferons, as well as elevated pro-inflammatory cytokines (IL-1β, IL-6, TNF-α, and IL-12p70) and IL-10, whereas neutralizing antibody (Neut. Ab.) titers against wild-type and variant strains, including Omicron, were comparable between groups. Immunophenotyping demonstrated preserved T- and B-cell subset distributions, except for reduced CD8+CD197+CD45RA (central memory) T cells in FV patients. ISG15 gene expression was upregulated in the T cells of FV patients. Correlation analyses linked IL-6 with disease activity and IL-8, GM-CSF, IFN-β, IL-10, and Alpha Neut. Ab. with organ damage. Complement C3 correlated inversely with IFN-α2 and IFN-γ, while C4 correlated positively with Alpha and Omicron Neut. Ab. These findings highlight that vaccination in SLE induces distinct interferon and cytokine signatures without consistent enhancement of neutralizing antibodies against SARS-CoV-2, underscoring the importance of integrated immune correlates in assessing vaccine responses in this population. Full article
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37 pages, 18599 KB  
Article
Diclofenac Immune-Mediated Hepatitis: Identification of Innate and Adaptive Immune Responses at Clinically Relevant Doses
by Jürgen Borlak and Reinhard Spanel
Int. J. Mol. Sci. 2025, 26(12), 5899; https://doi.org/10.3390/ijms26125899 - 19 Jun 2025
Cited by 1 | Viewed by 1853
Abstract
Diclofenac is an effective medication for pain and inflammation. However, its use has been linked to hepatitis. To gain insight into diclofenac’s ability to cause hepatitis, we investigated the regulation of major effectors of the immune system following daily treatment of minipigs at [...] Read more.
Diclofenac is an effective medication for pain and inflammation. However, its use has been linked to hepatitis. To gain insight into diclofenac’s ability to cause hepatitis, we investigated the regulation of major effectors of the immune system following daily treatment of minipigs at 3 and 15 mg/kg for 28 days. Histopathology evidenced lobular inflammation, and through a combination of immunogenomics and immunopathology, we detected marked innate and adaptive immune responses. We identified 109 significantly regulated genes linked to neutrophil, monocyte, Kupffer cell, and lymphocyte responses and 32 code for cytokine- and interferon-γ-signaling. In support of wound repair, immunopathology evidenced manifest upregulation of macrophage migration inhibitory factor and CD74. Furthermore, the strong expression of IgG and IgM underscored humoral immune responses. Diclofenac caused an activation of the complement system, especially the C1 inhibitor of the classical pathway and C3 with critical functions in liver regeneration. The marked expression of complement factor B and H of the alternate pathway modulated B-cell responses. Likely, the upregulation of factor H protected hepatocytes from injury by limiting complement-mediated damage of inflamed cells. Additionally, diclofenac treatment elicited marked hepatic expression of lysozyme and KLF6. The latter earmarks M1-polarized Kupffer cells. We observed an extraordinary induction of calprotectin/S100A9 and of the monocyte/macrophage CD163 scavenger receptor, and therefore, we detected innate immune sensing of damaged cells. Lastly, we noted an unprecedented induction of the acute phase reactant SAA1 and DEC-205, which recognize apoptotic and necrotic cells. Together, our results offer mechanistic insights into immune-mediated liver injury patterns following diclofenac treatment. Full article
(This article belongs to the Section Molecular Toxicology)
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15 pages, 2359 KB  
Article
The Effects of Ferulic Acid on the Growth Performance, Immune Function, Antioxidant Capacity, and Intestinal Microbiota of Broiler Chickens
by Xianguo Yi, Quanchao Ma, Zhili Li, Yuli Hu, Haigang Wu, Rui Wang, Xuyang Sun, Enen Wang, Chaofeng Ma and Qingmin Qin
Genes 2025, 16(5), 572; https://doi.org/10.3390/genes16050572 - 13 May 2025
Viewed by 1131
Abstract
Objectives: Ferulic acid is a natural and safe herbal feed additive. This study aims to evaluate the effects of ferulic acid on the growth performance, anti-inflammatory and antioxidant capacities, immune function, and intestinal microbiota of broiler chickens. Methods: A total of 320 broiler [...] Read more.
Objectives: Ferulic acid is a natural and safe herbal feed additive. This study aims to evaluate the effects of ferulic acid on the growth performance, anti-inflammatory and antioxidant capacities, immune function, and intestinal microbiota of broiler chickens. Methods: A total of 320 broiler chickens, aged 14 days, were randomly divided into four groups: a blank control group (MA group), a low-concentration ferulic acid group (BM group, 10 mg/kg), a medium-concentration ferulic acid group (CM group, 30 mg/kg), and a high-concentration ferulic acid group (DM group, 90 mg/kg) after a 14-day acclimatization period. The experiment lasted for 28 days, and the chickens were dissected on day 29. Results: The results showed that compared to the MA group, the feed-to-meat ratio in the CM and DM groups was significantly reduced. The activity of duodenal trypsin in the CM and DM groups was significantly enhanced, and the activity of pancreatic protease in the DM group was significantly increased. The serum levels of urea nitrogen, creatinine, and triglycerides were significantly elevated in the CM and DM groups. The serum malondialdehyde (MDA) levels in the BM, CM, and DM groups were significantly reduced, while the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were significantly increased in the CM and DM groups. The serum interleukin-2 (IL-2) levels in the BM group were significantly decreased, while interferon-gamma (IFN-γ) levels in the CM group and complement component 3 (C-3) levels in the DM group were significantly increased. The mRNA expression levels of TLR4, MyD88, NF-κB, TNF-α, NLRP3, IL-1β, and IL-18 in the jejunum of the DM group were significantly reduced. The diversity of cecal microbiota in the ferulic acid groups changed, with a certain degree of increase in the relative abundance of Spirulina and Ruminococcus. The relative abundance of Escherichia coli in the DM group significantly increased, altering the metabolic function of the cecal microbiota in broiler chickens. Conclusions: The above results indicate that ferulic acid, as a novel feed additive for broiler chickens, has an impact on the growth performance, anti-inflammatory and antioxidant capacity, immune function, and intestinal microbiota of broiler chickens. Full article
(This article belongs to the Special Issue Genetic Breeding of Poultry)
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18 pages, 5795 KB  
Article
C1QBP Modulates DNA Damage Response and Radiosensitivity in Hepatocellular Carcinoma by Regulating NF-κB Activity
by Haitao Zhou, Yanjin Wu, Jiahui Meng, Xiaotong Zhao, Yujia Hou, Qin Wang and Yang Liu
Int. J. Mol. Sci. 2025, 26(10), 4513; https://doi.org/10.3390/ijms26104513 - 9 May 2025
Viewed by 1160
Abstract
C1QBP (Complement Component 1 Q Subcomponent-Binding Protein) plays a critical role in maintaining cellular metabolism, but its function in radiation-induced damage remains unclear. In this study, we generated C1QBP-deficient Huh-7 hepatocellular carcinoma (HCC) cells using CRISPR/Cas9 technology and observed that C1QBP deficiency significantly [...] Read more.
C1QBP (Complement Component 1 Q Subcomponent-Binding Protein) plays a critical role in maintaining cellular metabolism, but its function in radiation-induced damage remains unclear. In this study, we generated C1QBP-deficient Huh-7 hepatocellular carcinoma (HCC) cells using CRISPR/Cas9 technology and observed that C1QBP deficiency significantly enhanced radiation-induced damage, as indicated by reduced cell proliferation, impaired colony formation, and increased γ-H2AX foci, a marker of DNA double-strand breaks. Additionally, C1QBP deficiency resulted in elevated phosphorylation of key DNA damage response (DDR) molecules, ATM and CHK2, and caused pronounced S phase cell cycle arrest. Mechanistic investigations revealed that C1QBP modulates NF-κB nuclear activity via the AMPK signaling pathway. The loss of C1QBP reduced NF-κB nuclear translocation, further exacerbating radiation-induced damage. Reintroducing C1QBP alleviated DNA damage, enhanced cell proliferation, and improved survival following radiation exposure. These findings highlight the critical role of C1QBP in modulating HCC cells radiosensitivity and underscore its potential as a therapeutic target to enhance radiotherapy outcomes. Full article
(This article belongs to the Special Issue Radiation-Induced DNA Damage and Toxicity)
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18 pages, 610 KB  
Article
Multiomics Analysis of a Micronutrient-Rich Dietary Pattern and the Aging Genotype 9p21 on the Plasma Proteome of Young Adults
by Sara Mahdavi, Katie Rosychuk, David J. A. Jenkins, Andrew J. Percy, Christoph H. Borchers and Ahmed El-Sohemy
Nutrients 2025, 17(8), 1398; https://doi.org/10.3390/nu17081398 - 21 Apr 2025
Cited by 1 | Viewed by 1872
Abstract
Background: Diet is one of the most significant modifiable lifestyle factors influencing human health, contributing to both morbidity and mortality. Genetic variations in the pleiotropic 9p21 risk locus further shape premature aging, disease susceptibility, and have been strongly linked to cardiovascular disease [...] Read more.
Background: Diet is one of the most significant modifiable lifestyle factors influencing human health, contributing to both morbidity and mortality. Genetic variations in the pleiotropic 9p21 risk locus further shape premature aging, disease susceptibility, and have been strongly linked to cardiovascular disease (CVD), metabolic disorders, certain cancers, and neurodegenerative conditions. However, given that this region was discovered based on Genome-Wide Association Studies, the mechanisms by which 9p21 exerts its effects remain poorly understood and its interactions with diet and biomarkers are insufficiently explored. Methods: This study investigated the association between the rs2383206 SNP in 9p21, dietary patterns, and plasma proteomic biomarkers in a multi-ethnic cohort of 1280 young adults from the Toronto Nutrigenomics and Health Study. Participants’ dietary intake was assessed using a validated food frequency questionnaire, and dietary patterns were categorized using principal component analysis. Plasma proteomics analyses quantified 54 abundant proteins involved in the cardiometabolic and inflammatory pathways. Genotyping identified individuals who were homozygous for the 9p21 risk allele (GG), known to confer the highest susceptibility risk to premature aging and multiple chronic diseases. Results: A significant interaction was observed between the 9p21 genotype and adherence to a micronutrient-rich Prudent dietary pattern for eight plasma proteins (α1 Antichymotrypsin, Complement C4 β chain, Complement C4 γ chain, Complement C9, Fibrinogen α chain, Hemopexin, and Serum amyloid P-component). However, only Complement C4-γ showed a pattern consistent with the risks associated with the 9p21 genotype and adherence to a Prudent diet. Individuals with the high-risk GG genotype had significantly higher concentrations of Complement C4-γ, but only among those with a low adherence to a Prudent diet. Conclusions: These findings suggest that Prudent dietary patterns rich in micronutrients may counteract genetic-mediated proinflammatory susceptibility by modulating key proteomic biomarkers in young adults, highlighting the potential for tailored dietary interventions to mitigate disease risk. This study also introduces a novel framework for post hoc micronutrient resolution within dietary pattern analysis, offering a new lens to interpret nutrient synergies in gene–diet interaction research. Full article
(This article belongs to the Special Issue Recent Advances in Nutrigenomics and Nutrigenetics)
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15 pages, 4925 KB  
Article
C3-Liposome Delivery of MUC1 Peptide and TLR Agonists Enhances Adaptive Immunity and Results in Sex-Based Tumor Growth Differences
by Shahab Soltani, Ameneh Arabi, Kristine Mann, Austin Hess, Holly A. Martinson and Max Kullberg
Pharmaceutics 2025, 17(4), 468; https://doi.org/10.3390/pharmaceutics17040468 - 3 Apr 2025
Viewed by 1130
Abstract
Background: Mucin-1 (MUC1) is a glycoprotein that is hypoglycosylated and overexpressed in most adenocarcinomas, making it a promising target for cancer vaccines. Our group previously demonstrated that C3 (OPSS)-liposomes enhance antigen uptake by antigen-presenting cells (APCs) via the complement C3 pathway and, [...] Read more.
Background: Mucin-1 (MUC1) is a glycoprotein that is hypoglycosylated and overexpressed in most adenocarcinomas, making it a promising target for cancer vaccines. Our group previously demonstrated that C3 (OPSS)-liposomes enhance antigen uptake by antigen-presenting cells (APCs) via the complement C3 pathway and, when combined with toll-like receptor (TLR) agonists, reduce tumor growth in murine cancer models. Methods: In the present study, we evaluate the immunogenicity of MUC1 peptide vaccines encapsulated in C3-liposomes, with and without TLR agonists, using MUC1-tolerant transgenic mice challenged with Lewis lung carcinoma (LLC.MUC1) cells. To assess vaccine effectiveness, tumor volumes were measured, and flow cytometry and ELISA and ELISPOT assays were used to assess the immune response. Results: Both male and female C57BL/6 transgenic mice vaccinated with MUC1 C3-liposomes developed significantly smaller tumors than those vaccinated with free MUC1 peptide or PBS. Notably, a sex-dependent response emerged in mice vaccinated with MUC1 C3-liposomes with TLR agonists (TLR4, TLR7/8, and TLR9); male mice exhibited greater tumor suppression than females. Flow cytometry analysis revealed that female mice had significantly higher levels of CD11b+, LY6C+, and LY6G+ MDSC cells, suggesting a potential mechanism for the sex difference. Additionally, MUC1 C3-liposome vaccination elicited robust adaptive immune responses, including significantly higher levels of IFN-γ-producing T cells and MUC1-specific IgG antibodies compared to non-encapsulated MUC1 or TLR adjuvant-only formulations. Conclusions: These findings underscore the potential of C3-liposome-based antigen vaccines to enhance anti-tumor immunity and highlight the impact of sex differences in vaccine efficacy. Full article
(This article belongs to the Special Issue Lipid Nanostructures as Drug Carriers for Cancer Therapy)
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15 pages, 4898 KB  
Article
Comparative Effect of GABA and 1-MCP in Maintaining Strawberry Fruit Quality During Cold Storage
by Mihaela Iasmina Madalina Ilea, Huertas María Díaz-Mula, Alba García-Molina, María Celeste Ruiz-Aracil, Christian Fernández-Picazo and Fabián Guillén
Horticulturae 2025, 11(4), 370; https://doi.org/10.3390/horticulturae11040370 - 30 Mar 2025
Cited by 4 | Viewed by 1788
Abstract
Strawberries (Fragaria x ananassa Duch.) are highly perishable fruits with a short postharvest shelf life, requiring effective preservation techniques. This study evaluates the efficacy of γ-aminobutyric acid (GABA) and 1-methylcyclopropene (1-MCP) in maintaining strawberry quality during cold storage. Freshly harvested strawberries were [...] Read more.
Strawberries (Fragaria x ananassa Duch.) are highly perishable fruits with a short postharvest shelf life, requiring effective preservation techniques. This study evaluates the efficacy of γ-aminobutyric acid (GABA) and 1-methylcyclopropene (1-MCP) in maintaining strawberry quality during cold storage. Freshly harvested strawberries were treated with different concentrations of GABA (1–20 mM), and a commercial 1-MCP concentration was suggested for strawberries (250 nL L⁻1) before being stored at 2 °C for up to 12 days. Different quality, physiological, and biochemical parameters were evaluated every 3 days after harvest. Results showed that both GABA and 1-MCP treatments effectively delayed fruit senescence. GABA demonstrated a higher effect on reducing weight loss, around 35.01% and 5.29% for 10 mM GABA and 1-MCP, respectively, compared to control fruit after 12 days at 2 °C. These substances also were effective in preserving firmness, but better maintenance was observed for 10 mM GABA than for 1-MCP (5.11 ± 0.43 and 3.49 ± 0.37 N, respectively) compared with control fruit after 12 days at 2 °C (2.56 ± 0.46 N). On the other hand, lower GABA concentrations (1–5 mM) and 1-MCP were particularly effective in delaying colour evolution and maintaining cell membranes and bioactive compounds such as polyphenols. In conclusion, as a postharvest treatment, GABA potentially offers an alternative or complement to 1-MCP in prolonging strawberry storability. These findings could contribute to developing sustainable strategies to reduce postharvest losses and improve strawberry marketability. Full article
(This article belongs to the Section Postharvest Biology, Quality, Safety, and Technology)
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26 pages, 4794 KB  
Article
Modulation of Macrophages TLR4-Mediated Transcriptional Response by Lacticaseibacillus rhamnosus CRL1505 and Lactiplantibacillus plantarum CRL1506
by Masahiko Suzuki, Ayelen Baillo, Leonardo Albarracin, Mariano Elean, Rodrigo Serda, Yoshihito Suda, Fu Namai, Keita Nishiyama, Haruki Kitazawa and Julio Villena
Int. J. Mol. Sci. 2025, 26(6), 2688; https://doi.org/10.3390/ijms26062688 - 17 Mar 2025
Cited by 2 | Viewed by 1377
Abstract
Lacticaseibacillus rhamnosus CRL1505 and Lactiplantibacillus plantarum CRL1506 increase the resistance of mice to Gram-negative pathogens infections. In this work, we advanced the characterization of the CRL1505 and CRL1506 immunomodulatory properties by evaluating their effect on the Toll-like receptor 4 (TLR4)-triggered immune response in [...] Read more.
Lacticaseibacillus rhamnosus CRL1505 and Lactiplantibacillus plantarum CRL1506 increase the resistance of mice to Gram-negative pathogens infections. In this work, we advanced the characterization of the CRL1505 and CRL1506 immunomodulatory properties by evaluating their effect on the Toll-like receptor 4 (TLR4)-triggered immune response in macrophages. We performed experiments in murine RAW 264.7 macrophages stimulated with lipopolysaccharide (LPS) to evaluate the transcriptomic changes induced by lactobacilli. These in vitro experiments were complemented with in vivo studies in mice to determine the effect of CRL1505 and CRL1506 strains on Peyer’s patches and peritoneal macrophages. Microarray transcriptomic studies and qPCR confirmation showed that the CRL1505 and CRL1506 strains modulated the expression of inflammatory cytokines and chemokines as well as adhesion molecules in LPS-challenged RAW macrophages, making the effect of L. rhamnosus CRL1505 more remarkable. Lactobacilli also modulate regulatory factors in macrophages. L. plantarum CRL1506 increased il10 and socs2 while L. rhamnosus CRL1505 upregulated il27, socs1, and socs3 in RAW cells, indicating a strain-specific effect. However, in vivo, both strains induced similar effects. Peyer’s patches and peritoneal macrophages from mice treated with lactobacilli produced higher levels of tumor necrosis factor (TNF)-α, interferon (IFN)-γ, interleukin (IL)-6, and colony stimulating factor (CSF)-3 after LPS stimulation. This effect would allow improved protection against pathogens. In addition, both lactobacilli equally modulated socs1 and socs2 expressions and IL-10 and IL-27 production in Peyer’s patches macrophages and socs3 and IL-10 in peritoneal cells. Furthermore, lactobacilli reduced the production of IL-1β, IL-12, CSF2, C-C motif chemokine ligand (CCL)-2, and CCL8 in LPS-challenged macrophages. This differential modulation of regulatory and inflammatory factors would allow minimal inflammatory-mediated tissue damage during the generation of the innate immune response. This work provides evidence that L. rhamnosus CRL1505 and L. plantarum CRL1506 modulate macrophages’ TLR4-mediated immunotranscriptomic response, helping to improve protection against Gram-negative bacterial infections. Full article
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24 pages, 11308 KB  
Article
Microbiomic and Metabolomic Insights into the Mechanisms of Alfalfa Polysaccharides and Seaweed Polysaccharides in Alleviating Diarrhea in Pre-Weaning Holstein Calves
by Jianan Zhao, Haoliang Tian, Xiaohui Kong, Danqi Dang, Kaizhen Liu, Chuanyou Su, Hongxia Lian, Tengyun Gao, Tong Fu, Liyang Zhang, Wenqing Li and Wei Zhang
Animals 2025, 15(4), 485; https://doi.org/10.3390/ani15040485 - 8 Feb 2025
Cited by 2 | Viewed by 1752
Abstract
Neonatal calves’ diarrhea, which can be severe enough to cause death, has a significant impact on the global cattle industry. In this study, alfalfa polysaccharides and seaweed polysaccharides were found to significantly improve the diarrhea condition in neonatal calves. To explore the underlying [...] Read more.
Neonatal calves’ diarrhea, which can be severe enough to cause death, has a significant impact on the global cattle industry. In this study, alfalfa polysaccharides and seaweed polysaccharides were found to significantly improve the diarrhea condition in neonatal calves. To explore the underlying mechanisms, further microbiomic and metabolomic analyses were conducted. This study investigated the impact of alfalfa polysaccharides and seaweed polysaccharides on growth performance, serum metabolites, gut microbiota, and metabolomics in neonatal Holstein calves. A total of 24 newborn calves were randomly assigned to three groups, with 8 calves per treatment group. The control (CON) group was fed a basal diet, the alfalfa polysaccharide (AP) group received a basal diet supplemented with alfalfa polysaccharides (4 g/calf/day), and the seaweed polysaccharide group (SP) received a basal diet supplemented with seaweed polysaccharides (4 g/calf/day). These polysaccharides were plant extracts. Compared to the CON group, the results indicated that SP significantly enhanced the body weight, height, chest circumference, and average daily gain of Holstein calves (p < 0.05), while also reducing the diarrhea rate and improving manure scoring (p < 0.05). Compared to the CON, AP also reduced the diarrhea rate (p < 0.05). In terms of serum biochemistry, supplementation with AP and SP increased serum alkaline phosphatase (ALP) and insulin-like growth factor 1 (IGF-1) levels compared to the CON group (p < 0.05). Both AP and SP elevated serum catalase (CAT) and Total Antioxidant Capacity (T-AOC) levels, indicating enhanced antioxidant status (p < 0.05). Regarding immune responses, supplementation with AP and SP significantly increased serum complement component 3 (C3) and immunoglobulin M (IgM) levels, while significantly reducing pro-inflammatory cytokines interleukin-18 (IL-18), tumor necrosis factor alpha (TNF-α), and interferon-gamma (IFN-γ) compared to the CON group (p < 0.05). Microbiota analysis revealed that AP modulated the abundance of Firmicutes, while SP influenced the abundance of Prevotella and Succiniclasticum. AP and SP differentially influenced intestinal metabolites compared to the CON group, leading to enrichment in pathways related to immunity, antibacterial, and anti-inflammatory functions. These pathways included the biosynthesis of alkaloids from ornithine, lysine, and nicotinic acid, glucocorticoid and mineralocorticoid receptor canothersis/antagonists, secondary metabolite biosynthesis, and alkaloid biosynthesis from histidine and purine, thus alleviating intestinal inflammation. Therefore, by supplementing with AP and SP, the diarrhea rate in calves was reduced, and the immune function of Holstein calves was enhanced, while simultaneously promoting a higher relative abundance of beneficial gut bacteria and suppressing the relative abundance of pathogenic bacteria. Additionally, gut pathways associated with immune response and inflammation were modulated by AP and SP. This study provided valuable insights and theoretical underpinnings for the use of AP and SP in preventing diarrhea in neonatal calves. Full article
(This article belongs to the Section Cattle)
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15 pages, 1040 KB  
Article
Characterization of Streptococcus pyogenes Strains from Tonsillopharyngitis and Scarlet Fever Resurgence, 2023—FIRST Detection of M1UK in Bulgaria
by Emma Keuleyan, Theodor Todorov, Deyan Donchev, Ani Kevorkyan, Radoslava Vazharova, Alexander Kukov, Georgi Todorov, Boriana Georgieva, Iskra Altankova and Yordanka Uzunova
Microorganisms 2025, 13(1), 179; https://doi.org/10.3390/microorganisms13010179 - 16 Jan 2025
Cited by 2 | Viewed by 3268
Abstract
Recently a resurgence of Streptococcus pyogenes infections has arisen, with concerns around the highly virulent M1UK lineage. Our aim was to characterize S. pyogenes, the immune responses it causes, and to determine the presence of the M1UK lineage in Sofia, [...] Read more.
Recently a resurgence of Streptococcus pyogenes infections has arisen, with concerns around the highly virulent M1UK lineage. Our aim was to characterize S. pyogenes, the immune responses it causes, and to determine the presence of the M1UK lineage in Sofia, Bulgaria. In our study, the infections were confirmed by culture testing or rapid antigen test. Identification was performed by MALDI-TOF and was followed up by antibiotic susceptibility testing (EUCAST). Virulence factors were identified using multiplex PCR and whole genome sequencing (WGS). Immune responses were measured through detection of serum complement levels, lymphocyte subsets, and cytokine profiling. Out of 82 children, 38 had scarlet fever and the rest had streptococcal pharyngitis. Strains were susceptible to penicillin (β-lactams), macrolides, clindamycin, tetracyclines, co-trimoxazole, fluoroquinolones, and linezolid. Superantigen profiles were identified: SpeA + SpeJ (45%), SpeC, and SpeI + SpeH (27.5% each). A novel Multilocus sequence typing (MLST) haplotype in the mutS gene (d90b) was found in four strains. The M1UK lineage was detected for the first time in Bulgaria. We observed an increase in complement fractions C3 and C4 and a decrease in T lymphocytes. A significant increase in the levels of IFN-γ, IL-6, and IL-10 with corresponding reduction in IL-17A were revealed. In conclusion, the studied S. pyogenes strains were characterized by their susceptibility to antibiotics and the predominance of SpeA superantigen; for the first time in Bulgaria the presence of M1UK and a novel SNP variation in the mutS gene (d90b) were found. A mixed pattern of pro- and anti-inflammatory immune responses in patients was observed. Full article
(This article belongs to the Section Medical Microbiology)
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34 pages, 5759 KB  
Article
Expression and Immune Response Profiles in Nile Tilapia (Oreochromis niloticus) and European Sea Bass (Dicentrarchus labrax) During Pathogen Challenge and Infection
by Ahmed A. Saleh, Asmaa Z. Mohamed, Shaaban S. Elnesr, Asmaa F. Khafaga, Hamada Elwan, Mohamed F. Abdel-Aziz, Asmaa A. Khaled and Elsayed E. Hafez
Int. J. Mol. Sci. 2024, 25(23), 12829; https://doi.org/10.3390/ijms252312829 - 28 Nov 2024
Cited by 9 | Viewed by 4025
Abstract
Nile tilapia (Oreochromis niloticus) and European sea bass (Dicentrarchus labrax) are economically significant species in Mediterranean countries, serving essential roles in the aquaculture industry due to high market demand and nutritional value. They experience substantial losses from bacterial pathogens [...] Read more.
Nile tilapia (Oreochromis niloticus) and European sea bass (Dicentrarchus labrax) are economically significant species in Mediterranean countries, serving essential roles in the aquaculture industry due to high market demand and nutritional value. They experience substantial losses from bacterial pathogens Vibrio anguillarum and Streptococcus iniae, particularly at the onset of the summer season. The immune mechanisms involved in fish infections by V. anguillarum and S. iniae remain poorly understood. This study investigated their impact through experiments with control and V. anguillarum- and S. iniae-infected groups for each species. Blood samples were collected at 1, 3, and 7 days post bacterial injection to assess biochemical and immunological parameters, including enzyme activities (AST and ALT), oxidative markers (SOD, GPX, CAT, and MDA), and leukocyte counts. Further analyses included phagocyte activity, lysozyme activity, IgM levels, and complement C3 and C4 levels. Muscle tissues were sampled at 1, 3, and 7 days post injection to assess mRNA expression levels of 18 immune-relevant genes. The focus was on cytokines and immune-related genes, including pro-inflammatory cytokines (TNF-α, TNF-β, IL-2, IL-6, IL-8, IL-12, and IFN-γ), major histocompatibility complex components (MHC-IIα and MHC-IIβ), cytokine receptors (CXCL-10 and CD4-L2), antimicrobial peptides (Pleurocidin and β-defensin), immune regulatory peptides (Thymosin β12, Leap 2, and Lysozyme g), and Galectins (Galectin-8 and Galectin-9). β-actin was used as the housekeeping gene for normalization. Significant species-specific responses were observed in N. Tilapia and E. Sea Bass when infected with V. anguillarum and S. iniae, highlighting differences in biochemical, immune, and gene expression profiles. Notably, in N. Tilapia, AST levels significantly increased by day 7 during S. iniae infection, reaching 45.00 ± 3.00 (p < 0.05), indicating late-stage acute stress or tissue damage. Conversely, E. Sea Bass exhibited a significant rise in ALT levels by day 7 in the S. iniae group, peaking at 33.5 ± 3.20 (p < 0.05), suggesting liver distress or a systemic inflammatory response. On the immunological front, N. Tilapia showed significant increases in respiratory burst activity on day 1 for both pathogens, with values of 0.28 ± 0.03 for V. anguillarum and 0.25 ± 0.02 for S. iniae (p < 0.05), indicating robust initial immune activation. Finally, the gene expression analysis revealed a pronounced peak of TNF-α in E. Sea Bass by day 7 post V. anguillarum infection with a fold change of 6.120, suggesting a strong species-specific pro-inflammatory response strategy. Understanding these responses provides critical insights for enhancing disease management and productivity in aquaculture operations. Full article
(This article belongs to the Special Issue Fish Immunology, 5th Edition)
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17 pages, 5693 KB  
Article
Expressions for Stress Concentration Factors for T-Joints of Hollow and Concrete-Filled Square Cross-Sections for In-Plane Axial and Bending Loads
by Liyong Gao, Lei Jiang, Xingzheng Wang, Sheng Gao, Hongxu Cui, Jun Liu and Hekuan Zhou
Symmetry 2024, 16(8), 1082; https://doi.org/10.3390/sym16081082 - 21 Aug 2024
Cited by 2 | Viewed by 1495
Abstract
In recent years, square hollow section (SHS) joints with concrete-filled square hollow section (CFSHS) chords have increasingly been used in truss bridges where the fatigue life reliability is a critical issue. In this paper, a finite element analysis was performed to investigate the [...] Read more.
In recent years, square hollow section (SHS) joints with concrete-filled square hollow section (CFSHS) chords have increasingly been used in truss bridges where the fatigue life reliability is a critical issue. In this paper, a finite element analysis was performed to investigate the SCFs in SHS-CFSHS T-joints under in-plane bending in the brace, axial force in the chord and in-plane bending in the chord. The finite element models were developed and validated with experimental results. Then a parametric study was conducted with a reasonable range of three key non-dimensional parameters, i.e., β (width ratio between brace and chord), 2γ (width–to–wall thickness ratio of the chord) and τ (wall thickness ratio between brace and chord). Consequently, the stress concentration factor formulae for the fatigue design of SHS-CFSHS T-joints were proposed through multiple regression analysis. For in-plane bending in the brace, the maximum stress concentration factors were found at lines B and C for thick-walled chords (2γ = 25.0), while the stress concentration factors at all the lines need to be checked for thin-walled chords (2γ < 16.0). Under axial force in the chord and in-plane bending in the chord, only stress concentration factors at lines C and D needed to be considered. A comparison of stress concentration factors between SHS-SHS and SHS-CFSHS joints showed reductions of 10~26% and 14~31% in stress concentration factors in SHS-CFSHS joints under axial force in the brace and in-plane bending in the brace, respectively. In addition, it showed a general increase in stress concentration factors in SHS-CFSHS joints under axial force and in-plane bending in the chord. This reduction is attributed to the reduction in chord face deformation benefiting from the in-filled concrete. Meanwhile, the stress concentration factors caused by loads in the chord are much lower than those caused by loads in the brace. This work complements earlier studies on SHS-CFSHS T-joints under axial force. Full article
(This article belongs to the Section Engineering and Materials)
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20 pages, 6094 KB  
Article
Effects of Acremonium terricola Culture on Lactation Performance, Immune Function, Antioxidant Capacity, and Intestinal Flora of Sows
by Zhirong Chen, Lixia Xiao, Qian Sun, Qiangqiang Chen, Weidong Hua and Jinzhi Zhang
Antioxidants 2024, 13(8), 970; https://doi.org/10.3390/antiox13080970 - 9 Aug 2024
Cited by 4 | Viewed by 1771
Abstract
This study aimed to determine the effects of different doses of Acremonium terricola culture (ATC) on lactation performance, immune function, antioxidant capacity, and intestinal flora of sows. Forty-five Landrace sows (3–6 parity) were randomly assigned to the following three treatments from 85 days [...] Read more.
This study aimed to determine the effects of different doses of Acremonium terricola culture (ATC) on lactation performance, immune function, antioxidant capacity, and intestinal flora of sows. Forty-five Landrace sows (3–6 parity) were randomly assigned to the following three treatments from 85 days of gestation to 21 days after farrowing: a control diet (CON, basal diet), a low-dose Acremonium terricola culture diet (0.2% ATC, basal diet + 0.2% ATC), and a high-dose Acremonium terricola culture diet (0.4% ATC, basal diet + 0.4% ATC). Compared with the CON group, the supplementation of 0.2% ATC increased the average daily milk yield of sows by 4.98%, increased milk fat, total solids, and freezing point depression on day 1 postpartum (p < 0.05), increased serum concentration of Triiodothyronine, Thyroxin, and Estradiol on day 21 postpartum (p < 0.05). Compared with the CON group, the supplementation of 0.4% ATC increased the average daily milk yield of sows by 9.38% (p < 0.05). Furthermore, the supplementation of 0.2% ATC increased serum concentration of IgG, IgM, and IFN-γ, CD4 on day 1 postpartum (p < 0.05) and increased serum concentration of immunoglobulin A ( IgA), immunoglobulin G (IgG), immunoglobulin M ( IgM), complement 3 (C3), cluster of differentiation 4 (CD4), cluster of differentiation 8 (CD8), interferon-γ (IFN-γ) on day 21 postpartum (p < 0.05), while the supplementation of 0.4% ATC reduced serum concentration of IL-2 on day 21 postpartum (p < 0.05). Moreover, the supplementation of 0.4% ATC significantly increased serum concentration of catalase (CAT) (p < 0.05). Additionally, the supplementation of ATC affected the relative abundance of the intestinal flora at different taxonomic levels in sows and increased the abundance of beneficial bacteria such as in the norank_f__Eubacterium_coprostanoligenes group, Eubacterium_coprostanoligenes group, and Lachnospiraceae_XPB1014 group of sows, while reducing the abundance of harmful bacteria such as Phascolarctobacterium and Clostridium_sensu_stricto_1. These data revealed that the supplementation of ATC during late gestation and lactation can improve lactation performance, immune function, antioxidant capacity, and the gut microbiota. Compared with supplementation of 0.4% ATC, 0.2% ATC enhances the levels of thyroid-related hormones, specific antibodies, and cytokines in serum, promotes the diversity of beneficial gut microbiota, beneficial bacteria in the intestine, reduces the population of harmful bacteria, and thereby bolsters the immunity of sows. Hence, 0.2% ATC is deemed a more optimal concentration. Full article
(This article belongs to the Special Issue Oxidative Stress in Livestock and Poultry—2nd Edition)
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