Search Results (264)

Objectives: This study investigated the antithrombotic properties of a fibrinolytic enzyme (CFE) purified from the culture supernatant of Coprinus comatus using a zebrafish thrombosis model. Methods: A phenylhydrazine-induced thrombosis model was employed to evaluate the in vivo thrombolytic efficacy and mechanisms of CFE. Results: CFE significantly attenuated thrombogenesis by inhibiting erythrocyte aggregation in the caudal vessels, reducing staining intensity (3.61-fold decrease) and staining area (3.89-fold decrease). Concurrently, CFE enhanced cardiac hemodynamics, increasing erythrocyte staining intensity (9.29-fold) and staining area (5.55-fold) while achieving an 85.19% thrombosis inhibition rate. Behavioral analysis confirmed improved motility, with CFE-treated zebrafish exhibiting 2.23-fold increases in total movement distance and average speed, alongside a 3.59-fold extension in active movement duration. Mechanistically, ELISA revealed the multi-pathway activity of CFE, promoting fibrinolysis through reductions in plasminogen, fibrinogen, and D-dimer; inhibiting platelet activation via downregulation of prostaglandin-endoperoxide synthase (PTGS), thromboxane A2 (TXA2), P-selectin, and von Willebrand factor (vWF); and modulating coagulation cascades through elevated protein C and tissue factor pathway inhibitor (TFPI) with concurrent suppression of coagulation factor VII (FVII). Conclusions: These results indicate that the fibrinolytic enzyme CFE, derived from Coprinus comatus, exerts potent antithrombotic effects, supporting its potential as a basis for fungal-derived natural antithrombotic functional food ingredients. Full article

Background/Objectives: Localized provoked vulvodynia (LPV) is characterized by chronic vulvar pain upon light touch to the vestibule, a specialized ring of tissue immediately surrounding the vaginal opening. LPV affects about 14 million people in the US, yet the etiopathology of the disease is unknown. In LPV, the vestibule expresses elevated levels of the pro-nociceptive pro-inflammatory mediators prostaglandin E₂ (PGE2) and interleukin-6 (IL-6), which corresponds to lower pain thresholds. Previous studies have shown reduced amounts of arachidonic acid (AA)-derived pro-resolving lipid mediators in tissue biopsies from LPV patients that might impede the resolution of inflammation. AA is obtained from dietary linoleic acid, pointing to a defect in the metabolism of dietary polyunsaturated fatty acids in LPV. We aimed to further explore the involvement of AA metabolism in LPV, which appears dysregulated in the vestibule of LPV patients and culminates in chronic inflammation and chronic pain. Methods: Vestibular and vulvar tissue biopsies obtained from LPV and non-LPV patients were used to generate fibroblast strains and assessed for COX/LOX expression using qRT-PCR. Fibroblast strains were treated with inflammatory stimuli, and then COX-1 and COX-2 expression was assessed using Western blot analysis. Pro-inflammatory mediator production was assessed using enzyme-linked immunosorbent assays (ELISAs). ALOX5 and ALOX12 expression was assessed using qRT-PCR. Finally, lipidomic analysis was carried out to screen for 143 lipid metabolites following inflammatory challenge. Results: Tissue and fibroblasts from LPV patients exhibited altered expression of COX/LOX enzymes and production of AA-derived lipid mediators compared to non-LPV patients. Conclusions: Lipid profiles of tissue and vestibular fibroblasts from LPV patients differed from non-LPV patients, and this difference was attributed to differential COX/LOX expression and activity, which metabolizes AA derived from dietary linoleic acid. This dysregulation fosters chronic inflammation and reduced resolution capacity in LPV patients, causing chronic pain. While further work is needed, these findings suggest that dietary modifications could impact the LPV mechanism. Full article

By stimulating living tissues with proper molecules, the angiogenesis and vasculogenesis processes can be observed. Prostaglandin E1 (PGE1), which is a molecule that widens blood vessels and which is used for several medical purposes, such as treating critical limb ischemia, is a typical leading molecule in angiogenesis studies. Nevertheless, its involvement in vasculogenesis and morphogenesis is a more specific subject in the field of developmental biology and therapeutic research. Vasculogenesis is the embryonic phenomenon in which endothelial progenitor cells generate new blood vessels. This phenomenon is distinct and divergent from angiogenesis, which entails the creation of novel blood vessels extending from pre-existing ones. Morphogenesis is the biological phenomenon responsible for the development of an organism or its components into a specific shape. Embryonic development and tissue regeneration are essential components. Current research is investigating the broader consequences of prostaglandins, such as PGE1, in the fields of developmental biology and regenerative medicine. Gaining knowledge about the impact of PGE1 on morphogenesis could provide valuable insights into congenital vascular abnormalities and innovative approaches for tissue repair and regeneration, especially in limb ischemia. In this study, a histologic and morphogenesis study was carried out on Artemia salina napi (first stage of development) by simulating the angiogenesis and morphogenesis processes using PGE1 as the top molecule with vasoactive properties and a series of benopyridyne (3-aminoquinolines, 5-amino quinolines, 8-aminoquinolines, 8-hydroxyquinolines and quinolines, respectively). A series of 30 Artemia salina napi were exposed to the compound listed before. Also, a lot of 30 unexposed Artemia salina napi was taken into account. In total, 210 Artemia salina napi were studied as a model for angionensis and morphogenesis. The study used wet experiments together with imaging reconstruction and graph-generating methodologies. The results show that PGE1 can initiate the shape of the vessel formation. Also, some quinoline series have a pro-mild morphogenetic and angiogenetic effect. Overall, PGE1 plays a significant role in mediating vasculogenesis and morphogenesis through its vasodilatory, anti-inflammatory, and pro-proliferative effects on endothelial cells. PGE1 is involved mainly in increasing the length of the vessel, while the number of vascular branching has an all-simulating general impact. However, the molecules with mild vasculogenic effects tend to develop more complex, limited vascular networks, having a more localized role in the angiogenetic process. Overall imaging and graph analysis showed significant and distinct properties of the vascular network-derived graph. Full article

Glioblastoma (GBM) is the most aggressive form of malignant gliomas, with the eicosanoid-synthesizing enzyme cyclooxygenase-2 (COX-2) playing a pivotal role in its progression via the COX-2/prostaglandin E2/4 axis. COX-2 upregulations in tumor cells induces a pro-inflammatory tumor microenvironment (TME), affecting the behavior of invading bone marrow-derived macrophages (Mϕ) and brain-resident microglia (MG) through unclear autocrine and paracrine mechanisms. Using CRISPR/Cas9 technology, we generated COX-2 knockout U87 glioblastoma cells. In spheroids and in vivo xenografts, this resulted in a significant inhibition of tumorigenic properties, while not observed in standard adherent monolayer culture. Here, the knockout induced a G1 cell cycle arrest in adherent cells, accompanied by increased ROS, mitochondrial activity, and cytochrome c-mediated apoptosis. In spheroids and xenograft models, COX-2 knockout led to notable growth delays and increased cell death, characterized by features of both apoptosis and autophagy. Interestingly, these effects were partially reversed in subcutaneous xenografts after co-culture with Mϕ, while co-culture with MG enhanced the growth-suppressive effects. In an orthotopic model, COX-2 knockout tumors displayed reduced proliferation (fewer Ki-67 positive cells), increased numbers of GFAP-positive astrocytes, and signs of membrane blebbing. These findings highlight the potential of COX-2 knockout and suppression as a therapeutic strategy in GBM, particularly when combined with suppression of infiltrating macrophages and stabilization of resident microglia populations to enhance anti-tumor effects. Full article

Cats are different from dogs, and many questions remain open concerning the establishment of pregnancy. In cats, as in dogs, no feto-maternal signaling leading to establishment of pregnancy is known. But as opposed to dogs, the placenta is a source of steroid hormones and corticotropin-releasing hormone (CRH). Scarce information is available on physiological mechanisms at the uterine level during early gestation; more studies are needed on lymphocyte subsets, feto-maternal crosstalk and other mechanisms leading to local immunosuppression, allograft acceptance and embryo nidation and invasion. Recent studies investigate the function of extracellular vesicles (EVs); however, there is no study on embryo- or endometrium-derived EV. During pregnancy, anti-Müllerian hormone (AMH) serum concentrations were found to be higher than in non-pregnant cats, and a recent study found that supraphysiological levels may lead to pregnancy loss; the function of AMH during pregnancy warrants investigation. Most information is available on corpus luteum development and function, showing some similarities to dogs. Some information on maintenance of feline pregnancy was obtained by ovariectomy (OE) or the use of endocrine disruptors, showing that OE does not lead to pregnancy loss in all cases, especially when performed after day 35; the variable effect is still not fully understood. Antiprogesterone, dopamine agonists and prostaglandins were used in different dosages and treatment schemes and showed variable effect during the second half of gestation, highlighting progesterone and prolactin as key hormones for the maintenance of gestation. Some events during early gestation are comparable with the canine species, even though they appear earlier, like the entrance of the zygote into the uterus and implantation; however, significant differences are present concerning the histomorphology of the placenta and, in a few cases, even the gross morphology as in some cats, where the zonary placenta does not completely surround the fetus. Sonographical monitoring of feline pregnancy requires knowledge of species-specific developmental steps and the differential appearance of fetal and maternal structures in comparison with dogs. Full article

Background/Objectives: Latanoprost is a leading active pharmaceutical ingredient belonging to the synthetic prostaglandin F2α analogs, widely used as a first-line treatment for open-angle glaucoma and increased intraocular pressure. This study addresses the critical need for an accurate and precise chromatographic method that is capable of simultaneously quantifying latanoprost and six latanoprost-related substances in antiglaucoma eye drops. This will be crucial for patient safety and treatment efficacy. This method enables the separation of latanoprost isomers, (15S)-latanoprost, latanoprost enantiomer, and 5,6-trans latanoprost from latanoprost signal. Furthermore, it is specific for the well-known latanoprost degradants—the major latanoprost acid and the minor 15-ketolatanoprost—as well as synthetic derivatives, such as triphenylphosphine oxide (TPPO) and propan-2-yl 5-(diphenylphosphoryl)pentanoate (IDPP). Using forced degradation studies using high temperatures, UV light, alkalis, acids, and oxidizing agents, the degradation profiles of the drugs were characterized and the method’s stability-indicating power was confirmed. Methods: Separation was achieved on a stationary combined system comprising chiral and cyano columns. Reverse-phase gradient elution and UV 210 nm detection were employed. The novel method was validated according to the European Medicines Agency International Council for Harmonisation Q2 Validation of analytical procedures—Scientific guideline. Results: The method was shown to be linear in the range of 40–60 µg/mL for latanoprost and 0.05–2.77 µg/mL for related substances, confirmed by a correlation coefficient of r = 0.999. Recoveries for latanoprost were obtained within the range of 98.0–102.0% for assays and 90.0–110.0% for impurities. The detection and quantification limits for latanoprost were 0.025 µg/mL and 0.35 µg/mL, respectively. Conclusions: The analytical procedure developed is adequately sensitive, precise, and accurate compared to existing methods. The method can be reliably used to control the critical quality attributes of low-dose latanoprost products, ensuring their required high pharmaceutical quality, which translates into improvements in patient care. This advancement holds significant implications for enhancing the therapeutic management of glaucoma, ensuring drug safety and efficacy. Full article

Coking activities produce high concentrations of aromatic compounds (ACs) and related substances, which may have impacts on human health. However, the health effects of these substances on humans exposed to coking sites have not been fully elucidated. A total of 637 people were recruited to participate in this cross-sectional study. Using multiple linear regression and Bayesian kernel machine regression, we investigated the relationships between the urinary parent or metabolite forms of ACs (including metabolites of PAHs and their derivatives, nitrophenols, and chlorophenols) and hepatorenal biomarkers (HRBs), including total bilirubin, aspartate aminotransferase/alanine aminotransferase, serum uric acid, creatinine, albumin/globulin, and urea. The HRBs adopted in this study can effectively represent the status of human liver and kidney function. Mediation analysis was performed to investigate the possible mediating relationship between ACs and HRBs using oxidative stress markers as mediators. Our study indicated that ACs were significantly associated with increases in TBIL, AST/ALT, A/G, and UA, as well as a significant decrease in Cr. UREA showed no association with ACs among coking workers. The oxidative stress markers 8-hydroxy-2’-deoxyguanosine, 8-iso-prostaglandin-F2α, and 8-iso,15(R)-prostaglandinF2α mediated the induction of ACs on TBIL. Our results suggest that AC exposure in coking workers may be associated with adverse changes in hepatorenal biomarkers. This study highlights the significant impact of ACs from coking activities on workers’ hepatorenal biomarkers, providing crucial evidence for health risk assessment and prevention in affected populations. Full article

Plasma lipids are essential components of biological systems, transported through interactions with proteins to maintain cellular functions. These lipids exist in various forms, such as fatty acids, glycerolipids, glycerophospholipids, sphingolipids, sterols, and prenol lipids, derived from dietary intake, adipose tissue, and biosynthesis. While the association between certain fatty acids and cardiovascular diseases has been widely recognized, polyunsaturated fatty acids (PUFAs) exhibit cardioprotective effects, reducing risks of arrhythmias and heart-related mortality. This is due to their role in the production of eicosanoids, which modulate inflammation. Chronic inflammation, particularly in obesity, is significantly influenced by fatty acids, with saturated fatty acids promoting inflammation and PUFAs mitigating it. Oxylipins, bioactive molecules derived from the oxidation of PUFAs, play crucial roles in immune regulation across various organisms, including plants, fungi, and bacteria. These molecules, such as prostaglandins, leukotrienes, and resolvins, regulate immune responses during infection and inflammation. The production of oxylipins extends beyond mammals, with fungi and bacteria synthesizing these molecules to modulate immune responses, promoting both defense and pathogenesis. This review delves into the multifaceted effects of oxylipins, exploring their impact on host and microbial interactions, with a focus on their potential for therapeutic applications in modulating infection and immune response. Full article

In ruminants, the survival and development of the conceptus are heavily dependent on the composition of the uterine lumen fluid (ULF), which is influenced by prostaglandins (PGs). However, the variations in underlying PG-mediated ULF remain unclear. Herein, cycling heifers received an intrauterine infusion of vehicle as a control (CON) or meloxicam (MEL) on days 12–14 of the estrous cycle. Then, the ULF was collected on day 15 and alternations in its protein and lipid levels were analyzed. The suppression of prostaglandins induced by meloxicam resulted in 1343 differentially abundant proteins (DAPs) and 59 differentially altered lipids. These DAPs were primarily associated with vesicle-mediated transport, immune response, and actin filament organization, and were mainly concentrated on the ribosome, complement and coagulation cascades, cholesterol metabolism, chemokine signal pathway, regulation of actin cytoskeleton and starch and sucrose metabolism. These differential lipids reflected a physiological metabolic shift as the abundance of cell membrane-related lipids was modulated, including an accumulation of triacylglycerols and reductions in lysophosphatidylcholines, hexosyl ceramides, ceramides, and sphingomyelins species. Integration analysis of the DAPs and differentially altered lipid metabolites revealed that glycerophospholipid metabolism and choline metabolism were the core pathways. These findings highlight the potential roles of prostaglandins in ULF, providing new insights into the contributions of prostaglandins in the development of the conceptus. Full article

New therapies are highly needed to stabilize remission in patients with acute myeloid leukemia (AML). This study investigates the value of dendritic cells derived from leukemic blasts (DC_leu) to enhance anti-leukemic immunity after T-cell-enriched mixed lymphocyte cultures (MLCs). We correlated induced anti-leukemic activity with patient data, including biological, clinical and prognostic factors. Additionally, we correlated the frequencies of DC/DC_leu and leukemic-specific T cells with the achieved anti-leukemic activity after MLC. We show that mature DC/DC_leu can be generated using the immunomodulating Kit-M, which contains granulocyte–macrophage colony-stimulating-factor (GM-CSF) and prostaglandin E₁ (PGE₁), without inducing blast proliferation from leukemic whole blood (WB) samples. Activated leukemia-specific immune and memory cells increased after MLC with Kit-M-pretreated WB, leading to improved blast lysis. Enhanced anti-leukemic activity positively correlated with the frequencies of generated DC/DC_leu, proliferating leukemic-specific T cells and memory T cells, but not with leukemic blast counts, hemoglobin levels or platelet counts at diagnosis. No correlation was found between improved blast lysis and patients’ prognostic data, including age, gender, ELN risk groups, disease stage and response to induction chemotherapy. These findings underscore the potential of DC/DC_leu to evoke robust immune responses and potential immunological memory against AML. Overall, this innovative approach could pave the way for the development of improved immunotherapeutic strategies that function in vivo. Full article

Background: Astragali radix is a traditional Chinese medicine with potential therapeutic effects on periodontitis; however, its underlying mechanisms require further investigation. Methods: We employed network pharmacology, molecular docking, molecular dynamics simulations, and in vitro experiments to explore the potential actions and mechanisms of Astragali radix in treating periodontitis. Results: A total of 17 compounds (including the most prevalent one, Kaempferol) from Astragali radix and 464 corresponding targets were identified, from which five major active ingredients were selected based on the drug-active ingredient and periodontitis gene network. Protein–protein interaction (PPI) network analysis identified the top ten core potential targets, seven of which possess suitable crystal structures for molecular docking. These include interleukin-6 (IL6), tumor necrosis factor (TNF), AKT serine/threonine kinase 1 (AKT1), interleukin-1β (IL1β), prostaglandin G/H synthase-2 (PTGS2), matrix metalloproteinase-9 (MMP9), and caspase-3 (CASP3). Additionally, 58 Gene Ontology (GO) terms and 146 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were identified. The five major active ingredients and seven core targets mentioned above were subjected to molecular docking analysis using Discovery Studio 2019 software. Molecular dynamic simulations confirmed a stable interaction between the CASP3 and the Kaempferol ligand system. In vitro experiments indicated that Kaempferol significantly inhibited lipopolysaccharide (LPS)-induced apoptosis in human periodontal ligament stem cells and reduced the expression levels of IL6, CASP3 and MMP9. Conclusions: This study systematically elucidates that the primary active ingredients derived from Astragali radix exert their pharmacological effects (including anti-inflammation and anti-apoptosis) primarily by interacting with multiple targets. These findings establish a promising foundation for the targeted application of Astragali radix in the treatment of periodontitis. Full article

Ethenzamide (2-ethoxybenzamide), besides acetylsalicylic acid, is one of the mostly used salicylic acid derivatives in pharmaceuticals. It has analgesic and anti-inflammatory effects that originate from the inhibition of cyclooxygenase (COX-1) activity, thus blocking prostaglandin synthesis. In this work, efficient and eco-friendly methods were developed for the synthesis of ethenzamide via the O-alkylation reaction of salicylamide. The reactions were carried out under conventional conditions in a solvent-free system using variant solvents and different phase transfer catalysts (PTC) in the presence of microwave radiation or ultrasonic conditions. It was shown that in solvent-free conditions using TBAB as a catalyst, ethenzamide can be obtained within 15 min at 80 °C with 79% yield. Meanwhile, using microwave radiation under the same conditions, the reaction time can be shortened to 90 s with 92% yield. Notably, high yields can be achieved under PTC in water (or organic solvent-free) conditions using microwave radiation (2 min, 94%) or ultrasound (10 min, 95% efficiency). The studies prove that the PTC synthesis process of ethenzamide can be conducted under mild conditions, with a shorter reaction time and remarkably lower energy consumption in comparison to conventional processes, thus actualizing “green chemistry” for practical ethenzamide preparation. Full article

The purpose of this review is to propose that lipoxin A4 (LXA4), derived from arachidonic acid (AA), a potent anti-inflammatory, cytoprotective, and wound healing agent, may be useful to prevent and manage diabetic retinopathy (DR). LXA4 suppresses inappropriate angiogenesis and the production of pro-inflammatory prostaglandin E2 (PGE2), leukotrienes (LTs), 12-HETE (12-hydroxyeicosatetraenoic acid), derived from AA by the action of 12-lioxygenase (12-LOX)) interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α), as well as the expression of NF-κB, inducible NO (nitric oxide) synthase (iNOS), cyclooxygenase-2 (COX-2), intracellular adhesion molecule-1 (ICAM-1), and vascular endothelial growth factor (VEGF)—factors that play a role in DR. Thus, the intravitreal injection of LXA4 may form a new approach to the treatment of DR and other similar conditions such as AMD (age-associated macular degeneration) and SARS-CoV-2-associated hyperinflammatory immune response in the retina. The data for this review are derived from our previous work conducted in individuals with DR and from various publications on LXA4, inflammation, and DR. Full article

Background/Objectives: Umckalin, a coumarin derivative abundantly present in the root extract of Pelargonium sidoides, is a key bioactive compound known for its antimicrobial, antiviral, antitubercular, and immunomodulatory properties. Its therapeutic potential has been extensively studied, particularly in the context of respiratory diseases. This study aimed to evaluate the potential of umckalin as a therapeutic agent for chronic inflammatory diseases and to elucidate its underlying mechanisms of action. Methods: Using lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages as an experimental model, we investigated the inhibitory effects of umckalin on inflammatory mediators and cytokine production. We measured levels of nitric oxide (NO), prostaglandin E₂ (PGE₂), tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and interleukin-1 beta (IL-1β), and assessed the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). Additionally, the regulation of MAPK signaling pathways, including JNK, p38 MAPK, and ERK, was analyzed. Results: The results demonstrated that umckalin significantly reduced the levels of NO, PGE₂, TNF-α, IL-6, and IL-1β in LPS-stimulated RAW 264.7 cells. Umckalin also suppressed the expression of iNOS and COX-2, leading to decreased NO and PGE₂ production. Furthermore, umckalin effectively regulated inflammatory responses by reducing the phosphorylation of MAPK signaling pathways, including JNK, p38 MAPK, and ERK. Conclusions: These findings indicate that umckalin inhibits the production of TNF-α, IL-6, IL-1β, and NO, while regulating MAPK signaling pathways, thereby suppressing the expression of iNOS and COX-2. This study highlights the potent anti-inflammatory effects of umckalin and suggests its potential as a promising candidate for the treatment of chronic inflammatory diseases. Full article

Background/Objectives: This study investigates the metabolic profile of a single dose of etodolac in healthy volunteers, focusing on pharmacokinetics, clinical parameters, and metabolomic variations to identify biomarkers and pathways linked to drug response, efficacy, and safety. Methods: Thirty-seven healthy volunteers, enrolled after rigorous health assessments, received a single dose of etodolac (Flancox^® 500 mg). Pharmacokinetic profiles were determined using tandem mass spectrometry analysis, and the metabolomic profiling was conducted using baseline samples (pre-dose) and samples at maximum drug concentration (post-dose) via liquid chromatography coupled with a quadrupole time-of-flight mass spectrometer. Network analysis was employed to interpret the data. Results: Correlations were observed between metabolomic profiles and pharmacokinetic parameters as well as clinical characteristics. Notably, metabolites derived from arachidonic acid, such as prostaglandins and leukotrienes, were linked to etodolac’s pharmacokinetics. Other metabolites involved in pathways like cholesterol biosynthesis, bile salts, riboflavin, and retinoic acid signaling were correlated with hematological and liver function parameters. These findings are consistent with the infrequent adverse events reported by participants, including hematological and biochemical changes in liver function. Conclusions: A set of metabolites was identified in possible associations between specific pathways and unusual side effects, comparing the metabolic profiles before and after doses of etodolac. Our results highlight the importance of optimizing drug therapy and minimizing adverse events by taking into account individual metabolic profile information. Full article