Search Results (1,256)

Previous research has demonstrated that the transient bilateral common carotid artery occlusion and reperfusion (BCCAO/R) effectively models early brain inflammation resulting from sudden hypoperfusion and subsequent reperfusion. According to studies showing that diet and nutrition strongly influence brain neuroplasticity, in this study we evaluated whether kaempferol (KAM), a dietary flavonoid, offers neuroprotection in a rat BCCAO/R model. Adult Wistar rats were gavage fed a single dose of KAM (40 mg) six hours before surgery. Comprehensive lipidomic and molecular analyses were conducted on samples from the frontal and temporal-occipital cortices, as well as the plasma. In the frontal cortex, KAM elevated anti-inflammatory N-acylethanolamines palmitoylethanolamide (PEA), oleoylethanolamide (OEA), and docosahexaenoylethanolamide (DHAEA) and reduced oxidized arachidonic acid metabolites. KAM also downregulated cyclooxygenase- 2 (COX-2) protein and selectively decreased the endocannabinoid 2-arachidonoylglycerol (2-AG), showing a shift in AA metabolism. These molecular changes correlated with increased levels of peroxisome proliferator-activated receptor alpha (PPARα) and cannabinoid receptors CB1R and CB2R, supporting activation of both nuclear and membrane-bound anti-inflammatory pathways. No significant changes were observed in the temporal-occipital cortex. In plasma, DHAEA levels increased similarly to those in the cortex. However, rises in PEA and OEA were detected only in sham-operated KAM-treated animals, suggesting possible central redistribution under hypoperfusion/reperfusion stress. In summary, these findings demonstrate that KAM exerts dual anti-inflammatory effects by inhibiting COX-2-mediated prostanoid synthesis and promoting PPARα-driven lipid signaling. This dual mechanism highlights the potential of KAM as a dietary intervention to reduce neuroinflammation associated with hypoperfusion–reperfusion challenges. Full article

The ban on antibiotic growth promoters in livestock has intensified the search for effective probiotic alternatives. This study assessed the impact of a novel probiotic cocktail—comprising Lactobacillus plantarum AMT14 and AMT4, L. rhamnosus AMT15, and Bifidobacterium animalis AMT30—on the serum metabolome of lambs using an untargeted GC/MS approach. Sixteen Kamieniec lambs were divided into control and probiotic groups, with serum collected on days 0, 15, and 30. Metabolomic profiling revealed significant alterations in lipid and amino acid metabolism in the probiotic group. By day 15, 38 metabolites were upregulated, including 9,12-octadecadienoic acid, arachidonic acid, and cholesterol. On day 30, key increases included D-glucose, oleic acid, glycine, decanoic acid, and L-leucine. Multivariate analyses (PCA, PLS-DA) demonstrated clear separation between groups, and ROC analysis identified strong biomarkers with high predictive accuracy. These results suggest that probiotic supplementation can beneficially modulate host metabolism, potentially enhancing immune and physiological function in lambs. This highlights the value of multi-strain LAB-Bifidobacterium probiotics as a promising strategy for improving health and reducing antibiotic reliance in ruminant production systems. Full article

Background: Pathological pregnancy is associated with various complications that may affect the health of both the mother and her offspring. In recent years, lipid metabolites such as hydroxyeicosatetraenoic (HETE) fatty acids and hydroxyoctadecadienoic (HODE) fatty acids have gained increasing interest as potential biomarkers of pathological processes in pregnancy. The aims of the present study were to investigate changes in HETE and HODE levels during pathological pregnancy and to assess their potential role in the development and monitoring of pregnancy complications. Attempts were made to determine associations in cross-sectional studies and relationships in longitudinal ones. Methods: In this study, a liquid chromatograph (HPLC) was used to separate the eicosanoids. The study group consisted of 72 Caucasian women, divided into a control group (n = 51) and a group with non-physiological pregnancy (n = 21). Results: The study results show that the levels of the tested metabolites of the cyclooxygenase (COX) and lipoxygenase (LOX) pathways increased as pregnancy progressed. Women with non-physiological courses of pregnancy who developed gestational diabetes and/or preeclampsia were characterized by dysregulation of the inflammatory signaling processes involving eicosanoids. Conclusions: Carbohydrate abnormalities during pregnancy were mainly associated with increased synthesis of 5-oxoETE and the use of 5-HETE in the control group but were not visible in the diabetic group. HODE acids probably do not play a significant role in pathological pregnancy. The relatively small size of the pathological group and the wide range of gestational age mean that the tests should be standardized and carried out on a larger scale. Full article

The voltage-gated proton channel (H_v1) selectively transports protons (H⁺) across biological membranes in response to membrane potential changes. H_v1 is assembled as a dimer, and unlike most voltage-gated ion channels, it lacks a traditional central pore domain; instead, the voltage-sensing domain (VSD) of each monomer facilitates proton conduction via a hydrogen-bond network. H_v1 is widely expressed in various human cell types (e.g., immune cells, sperm, etc.) including tumor cells. In tumor cells, the accumulation of acidic intermediates generated by glycolysis under hypoxic conditions or ROS production leads to significant cytosolic acidification. H_v1 can remove protons from the cytosol rapidly, contributing to the adaptation of the cells to the tumor microenvironment, which may have significant consequences in tumor cell survival, proliferation, and progression. Therefore, H_v1 may be very promising not only as a tumor marker but also as a potential therapeutic target in oncology. Molecules that modulate the proton flux through H_v1 can be divided into two broad groups: inhibitors and activators. H_v1 inhibitors can be simple ions, small molecules, lipids, and peptides. In contrast, fewer H_v1 activators are known, including albumin, NH29, quercetin, and arachidonic acid. The mechanism of action of some inhibitors is well described, but not all. H_v1 modulation has profound effects on cellular physiology, especially under stress or pathological conditions, like cancer and inflammation. The therapeutic application of selective H_v1 inhibitors or activators could be a very promising strategy in the treatment of several serious diseases. Full article

Plantsterol esters (PSEs) exert beneficial effects on animal product quality, indicating their potential as a nutritional intervention strategy. This study investigated the effects of dietary PSE supplementation on lactation performance and lipid metabolism in Tarim Bactrian camels (Camelus bactrianus) to establish a scientific basis for its application in their husbandry. Thirty-two mid-lactation female camels were randomly allocated to four groups (n = 8): CON (basal diet), L (200 mg/kg PSE), M (400 mg/kg PSE), and H (800 mg/kg PSE). Since lactation performance is closely linked to metabolic status, biochemical and lipidomic analyses were conducted on serum and milk samples. Analysis revealed that the H group showed significantly increased milk yield, lactose yield, and milk fat yield compared to other groups. Serum cholesterol levels decreased progressively with higher PSE supplementation, while serum urea levels rose dose-dependently. Blood Glu was lower in the L group but higher in the H group relative to CON. Lipidomic profiling identified 644 and 257 differential metabolites in milk and serum, respectively. Milk metabolites were enriched in the EGFR inhibitor resistance, MAPK, and ErbB signaling pathways; serum metabolites were linked to glycerophospholipid, arachidonic acid, and linoleic acid metabolism. These findings indicate that PSE-modulated metabolites in serum and milk significantly influence lactation performance and glucolipid metabolism in Tarim Bactrian camels, supporting further investigation into precision nutrition strategies. Full article

Lipopolysaccharide (LPS) triggers oxidative damage in sheep hepatocytes, linked to ferroptosis. Ferulic acid (FA) is known for its antioxidative properties, but its protective role against LPS via ferroptosis regulation was unclear. The objective of this research is to explore the protective role of FA in mitigating LPS-induced oxidative stress in sheep hepatocytes. The experimental setup consisted of three groups: a control group, an LPS group treated with 10 µg/mL of LPS, and FA group that received both 10 µg/mL of LPS and 750 µg/mL of FA. We found that FA treatment decreased in contents of MDA and LDH. Metabolomics revealed that LPS affected glycerophospholipid metabolism, unsaturated fatty acids biosynthesis, ferroptosis, and arachidonic acid metabolism mainly by reducing the level of PUFAs and LPC in the hepatocyte supernatant, while FA affected glutathione metabolism by increasing L-cysteine, L-ornithine, L-glutamic acid, and L-glutamine. Moreover, transcriptomics demonstrated that the comparison of LPS and control groups were mainly enriched in arachidonic acid metabolism, glycerophospholipid metabolism, and ferroptosis, the comparison of FA and LPS groups was mainly enriched in glutathione metabolism. The results further confirmed the findings in the metabolomics and transcriptomics analyses, showing that LPS treatment upregulated the mRNA expression of ACSL4, LPCAT3, ALOX15, STEAP3, GPX4, GCLC, and GCL in hepatocytes, while reducing GSH and GR levels. In contrast, FA intervention attenuated LPS-induced iron overload by decreasing Fe²⁺ accumulation and suppressing the mRNA expression of ACSL4, LPCAT3, STEAP3, and ALOX15. Furthermore, FA enhanced the expression of GPX4, GCLC, GCLM, and restored GSH content in LPS-exposed hepatocytes. The above results demonstrated that the protective effect of FA on LPS-induced oxidative damage in the sheep hepatocytes was achieved by activating the GSH-GPX4 pathway and inhibiting lipid metabolism-mediated ferroptosis. Full article

Background/Objectives: Marine-derived oils rich in long-chain polyunsaturated fats have long been associated with positive effects on plasma lipid levels and anti-inflammatory responses. Abalone viscera are rich in oils that are rarely extracted and made available. Methods: Abalone viscera oil (AVO) was extracted by multistage countercurrent extraction using ethanol as a solvent, and its oil quality, fatty acid composition, and in vitro antioxidant activity were determined. Meanwhile, the anti-hyperlipidemic effect of AVO on HFD-induced hyperlipidemia mice was evaluated. Results: The abalone viscera were extracted at a solid–liquid ratio of 1:3 with an oscillation frequency of 300 rpm for 40 min, and the extraction rate was 81.18% after four-stage countercurrent extraction. The acid value, iodine value, peroxide value, vitamin E, and astaxanthin of AVO were 1.26 mg KOH/g, 140.9 g/100 g, 3.6 meq/kg, 105 mg/kg, and 533.8 mg/kg, respectively. The unsaturated fatty acids of AVO account for 56.60%, with eicosapentaenoic acid (C20:5n3) and arachidonic acid (C20:4n6) the two predominant PUFAs, and oleic acid (C18:1n9) the most dominant MUFA. The DPPH, ABTS, and ·OH radicals scavenging capacities of AVO increased with concentration, and the IC₅₀ values were 6.30 mg/mL, 0.45 mg/mL, and 8.95 mg/mL, respectively. Moreover, the administration of AVO significantly alleviated HFD-induced weight gain, liver fat accumulation, lipid disorder, and oxidative stress in mice. Conclusions: Collectively, our study provides a theoretical basis for the application of AVO and the comprehensive utilization of abalone viscera, which helps increase the additional value of abalone. Full article

Arachidonic acid (eicosatetraenoic acid) is the precursor of the eicosanoids, which include prostaglandins (PG). Methods based on GC-MS/MS are the Gold Standard for the quantitative analysis of eicosanoids in biological samples. After extraction and derivatization, biological F₂-prostaglandins are analyzed on quadrupole GC-MS/MS apparatus as pentafluorobenzyl (PFB) ester trimethylsilyl (TMS) ether derivatives, i.e., PFB-TMS. Negative-ion chemical ionization (NICI) in the ion source generates abundant anions due to [M-PFB]⁻, which are detected in the selected ion monitoring (SIM) mode. Collision-induced dissociation (CID) of [M-PFB]⁻ in the collision cell generates numerous product ions, which are suitable candidates for quantitative analyses in the selected reaction monitoring (SRM) mode. In this article, we report on investigations of gas-phase reactions of PFB-TMS derivatives of F₂-prostaglandins, which consist of PGF_2α, 8-iso-PGF_2α, and up to 62 further isomers, known as the F₂-isoprostanes. We performed a meta-analysis of previously reported CID mass spectra (32 eV) of PFB-(TMS)₃ of seven chemically closely related isomeric F₂-prostaglandins of the 15-F_2t-IsoP type. This unique dataset contains 19 product ions generated by CID of the common precursor at m/z 569 [M-PFB]⁻ in the m/z range of 150–600. All isomers produced the same product ions, which, however, greatly differed in their intensity. Principal Component Analysis (PCA) and Receiver Operating Characteristic (ROC) Analysis (ROCA) were performed. Two compounds, i.e., 8-iso-9β,11α-PGF_2α and 9α,11β-PGF_2α, and two product ions, i.e., m/z 299 [M-PFB-3×TMSOH]⁻ and m/z 215 [M-PFB-3×TMSOH-C₄H₈-C₂H₄]⁻, were noticeable. ROCA revealed the highest disagreement between PGF_2α and 8-iso-9β,11α-PGF_2α (AUC = 0.7075 ± 0.0834, p = 0.0248). PCA and ROCA are of limited value in the GC-MS/MS of closely chemically related F₂-prostaglandins. Fragmentation mechanisms were proposed for the formation of all 19 product ions generated by CID of common precursor anions due to [M-PFB]⁻. Full article

Patent ductus arteriosus (PDA) constitutes a significant clinical condition, frequently associated with a spectrum of complications that may profoundly compromise the health status of neonates, particularly those born preterm. Multiple predisposing factors—including prematurity, low birth weight, and respiratory insufficiency—have been consistently documented in the scientific literature. In this study, we investigated the influence of genetic polymorphisms in genes associated with the arachidonic acid–prostaglandin metabolic pathway. Specifically, we analyzed polymorphisms in genes encoding phospholipase A2 (rs10798059, rs1549637, rs4375, rs1805017, rs1051931), cyclooxygenase-1 (rs1236913), prostaglandin synthase 2 (rs13283456), and the prostaglandin E2 receptor EP4 (rs4613763). The study cohort comprised 99 preterm neonates born between 24 and 32 weeks of gestation. Genetic analyses were performed to identify polymorphisms in the aforementioned genes. Statistical evaluation demonstrated that selected polymorphic were significantly associated with an increased risk of patent ductus arteriosus development. This study represents a preliminary step toward elucidating the contribution of genetic variability to the pathogenesis of patent ductus arteriosus. Improved understanding of these molecular mechanisms may facilitate the early identification of neonates at increased risk and support the implementation of targeted monitoring and preventive strategies in this high-risk population. Full article

Genetic engineering of macrophages, particularly for chimeric antigen receptor macrophage (CAR-M) therapy, holds great promise for immunotherapy, yet is constrained by the challenge of efficient gene delivery into primary macrophages, which are notoriously resistant to transfection. While conventional strategies focus on optimizing the physicochemical properties of lipid nanoparticles (LNP), they often fail to overcome the intrinsic biological barriers of these cells. Here, we introduced a “bioactive nanocarrier” paradigm, hypothesizing that incorporating a cellular modulator directly into LNP structure can synergistically overcome these barriers. We designed and synthesized a novel LNP by integrating the pro-inflammatory fatty acid, arachidonic acid (ARA), as a functional structural component (ARA-LNP). Systematic optimization of the ARA content and mRNA payload revealed a formulation that achieves high transfection efficiency (83.76%) in primary M2-polarized bone marrow-derived macrophages (BMDMs), a cell type that recapitulates pro-tumoral phenotype in the tumor microenvironment. Leveraging this advanced delivery platform, we successfully generated HER2-targeting CAR-M that demonstrated potent and specific phagocytic activity against HER2-expressing tumor cells in vitro. This work presents a powerful strategy where the nanocarrier itself transiently modulates the target cell state to enhance gene delivery, providing a new design principle for engineering macrophages and other hard-to-transfect immune cells for therapeutic applications. Full article

Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is caused by pathogenic mutations in the nuclear TYMP gene, which encodes the cytosolic enzyme thymidine phosphorylase. In addition to the systemic accumulation of thymidine and deoxyuridine, several case studies have reported abnormalities in a range of other metabolites in patients with MNGIE. Since metabolites are intermediates or end-products of numerous biochemical reactions, they serve as highly informative indicators of an organism’s metabolic activity. This study aimed to perform an untargeted metabolomic profiling to determine whether individuals with MNGIE exhibit a distinct plasma metabolic signature compared to 15 age- and sex-matched healthy controls. Metabolites were profiled using Ultra-High-Performance Liquid Chromatography–Mass Spectrometry (UHPLC-MS). A total of 160 metabolites were found to be significantly upregulated and 260 downregulated in patients with MNGIE. KEGG pathway enrichment analysis revealed disruptions in 20 metabolic pathways, with arachidonic acid metabolism and bile acid biosynthesis being the most significantly upregulated. Univariate receiver operating characteristic (ROC) curve analyses identified 23 individual metabolites with diagnostic potential, each showing an area under the curve (AUC) ≥ 0.80. We propose that an impaired resolution of inflammation contributes to a chronic inflammatory state in MNGIE, potentially driving disease progression. Additionally, we suggest that the gut–liver axis plays a central role in MNGIE pathophysiology, with hepatic function being bidirectionally influenced by gut-derived factors. Full article

Ischemic stroke is a leading cause of mortality and disability in adults. The inflammatory cascade is driven by various inflammatory molecules, such as interleukin-1β (IL-1β), and counteracted by its antagonist, interleukin-1 receptor antagonist (IL-1Ra). Eicosanoids are inflammatory derivatives of free fatty acids. Arachidonic acid (AA) derivatives exhibit pro-inflammatory activity, while eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) derivatives, known as specialized pro-resolving mediators, have anti-inflammatory properties. This study aimed to analyze potential associations between eicosanoids and key inflammatory molecules, including IL-1β and its antagonist IL-1Ra. In this prospective study, we investigated inflammatory molecules in 73 ischemic stroke patients. We analyzed interactions between IL-1β, IL-1Ra, and eicosanoids as follows: resolvin E1, prostaglandin E2, resolvin D1, lipoxin A4 (5S, 6R, 15R), protectin DX, maresin 1, leukotriene B4, 18RS-HEPE, 13S-HODE, 9S-HODE, 15S-HETE, 17 HDHA, 12S-HETE, 5-oxo-ETE, and 5-HETE. In 73 ischemic stroke patients, mean IL-1β was 1.31 ± 1.54 pg/mL and IL-1Ra 810.8 ± 691.0 pg/mL. Spearman correlations showed positive associations between IL-1β and protectin DX (ρ = 0.56, p < 0.001), and 17 HDHA (ρ = 0.26, p < 0.05) and 5-oxo-ETE (ρ = 0.27, p < 0.05). IL-1Ra correlated negatively with protectin DX (ρ = −0.58, p < 0.001) and 17 HDHA (ρ = −0.29, p < 0.05), and positively with leukotriene B4 (ρ = 0.34, p < 0.005). After multivariable adjustment, associations with IL-1β lost statistical significance, whereas the inverse relationships between IL-1Ra and protectin DX/17 HDHA remained significant (p < 0.005). Despite the known anti-inflammatory roles of protectin DX and 17 HDHA, and the pro-inflammatory role of leukotriene B4, their activity in the early subacute phase of ischemic stroke appears to be influenced by complex interplays, possibly mediated by IL-1β and IL-1Ra. The activity of protectin DX, 17 HDHA, and leukotriene B4 is correlated with IL-1β and IL-1Ra levels in the early subacute phase of stroke. Full article

Background: Physical activity induces the production of reactive oxygen species (ROS), which, at moderate levels mediate beneficial physiological adaptations, including insulin sensitivity and enhanced antioxidant defense. However, excessive ROS production during intense exercise may exceed endogenous antioxidant capacity, leading to oxidative stress and muscle damage. Objective: This study examined the effects of 30-day high-dose omega-3 fatty acid supplementation (9 g/day) on plasma fatty acid composition and the activity of antioxidant enzymes in recreational (n = 11) and trained (n = 10) runners, with emphasis on group- and time-specific responses. Methods: Plasma levels of arachidonic acid (AA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) were assessed at three time points: pre-, during, and post-supplementation period. Enzymatic activities of glutathione peroxidase (GPx), superoxide dismutase (SOD), and catalase (CAT) were measured at six time points, including before and after exercise sessions involving a 2800 m run followed by a 400 m sprint. Results: Omega-3 supplementation increased plasma EPA and DHA. In trained runners, it was associated with a transient reduction in GPx and a pronounced mid-phase decline in SOD, whereas enzyme activities remained stable in recreational runners. CAT activity did not change significantly in either group. Conclusions: Short-term high-dose omega-3 supplementation modulates antioxidant enzyme activity in a group- and time-dependent manner. The observed downregulation of GPx and SOD in trained runners may reflect altered redox signaling; however, its relevance for exercise performance remains uncertain. Further studies are warranted to elucidate the physiological and functional consequences of these findings. Full article

Hyperuricemia (HUA) is a metabolic disorder characterized by abnormal purine metabolism within the body. Ampelopsis japonica (Thunb.) Makino has traditionally been utilized in the treatment of various kidney diseases; however, its specific anti-hyperuricemic effects and the underlying mechanisms warrant further investigation. This study investigates the mechanism of action by which A. japonica extract (AJE) addresses HUA using a combination of pharmacology techniques, including network pharmacology and metabolomics. A HUA mouse model was established using potassium oxonate and hypoxanthine. AJE intervention significantly reduced serum uric acid and creatinine levels in HUA mice and markedly decreased glomerular atrophy and renal tubular degeneration. Metabolic profiling revealed distinct metabolic profiles between AJE-intervention and control groups, further demonstrating that AJE corrected disruptions in arginine biosynthesis, purine metabolism, pyrimidine metabolism, and arachidonic acid metabolism. The results of the network pharmacology-based study indicate that AJE can alleviate HUA by modulating the TNF pathway and the Toll-like receptor pathway. The mechanisms of action of AJE in HUA involve the inhibition of xanthine oxidase (XOD) to reduce uric acid synthesis, downregulation of URAT1 and GLUT9 to decrease uric acid reabsorption, and suppression of the TLR4/NF-κB pathway to mitigate inflammation in the HUA mouse model. Therefore, AJE demonstrates significant potential as a therapeutic intervention for HUA and its associated renal complications. Full article

The research aimed to examine the effects of fermentation and enzymatic hydrolysis of cottonseed protein on body weight changes, serum biochemistry, rumen function, intestinal health, and liver metabolism of suckling lambs. A total of twelve 7-day-old healthy male Hu sheep body weights (5.27 ± 0.48 kg) were randomly distributed into two groups. Starter feed regimens containing microbial fermentation of cottonseed protein (MFCP) or enzymatic hydrolysate of cottonseed protein (EHCP) were administered to lambs during the initial 60-day period. Results showed that compared with EHCP group, the serum glucose, ruminal acetic, propionic, butyric and valeric acids concentrations, jejunal immunoglobulin G content and mRNA expressions of Claudin 1 and Occludin, as well as the relative abundance of actinobacteriota and pseudoscardovia in the rumen were significantly increased in the MFCP group (p < 0.05), whereas an opposite trend was observed in the jejunum. α-amylase and trypsin enzymatic activities were observed between the two groups. Relative to EHCP treatment, the MFCP group exhibited 69 elevated and 103 reduced hepatic metabolites, and these metabolites displayed distinct enrichment patterns within specific metabolic networks, including fructose and mannose metabolism (p = 0.003), arachidonic acid metabolism (p = 0.017), glycerophospholipid metabolism (p = 0.036), and the cAMP signaling pathway (p = 0.047). Overall, microbial fermentation of cottonseed protein may be beneficial for strengthening intestinal barrier function and facilitating hepatic lipid metabolism and immune regulation, while enzymatic hydrolysis of cottonseed protein enhances gastrointestinal digestive enzyme activity, thereby promoting nutrient digestion of suckling lambs. Full article