Search Results (148)

The present study investigated the association between single-nucleotide polymorphisms (SNPs) and resistance to Micropterus salmoides rhabdovirus (MSRV) in largemouth bass. Candidate SNPs were first screened from transcriptome sequencing data and subsequently validated, resulting in 37SNP sites with good PCR amplification performance. Genotyping was performed using high-resolution melting (HRM) analysis, and associations between SNP loci and MSRV resistance were assessed using the chi-square test. Transcriptome analysis further elucidated immune-related pathways activated in response to MSRV infection. Among the tested loci, after Bonferroni correction, four SNPs remained highly significant (p < 0.001) and three became suggestive (p < 0.05). These markers provide valuable resources for marker-assisted selection in largemouth bass breeding programs and contribute to the development of disease-resistant strains. Full article

This study provides a comprehensive evaluation of the chemical composition and the biological properties of Reseda alba L., commonly known as white mignonette. Extracts obtained from leaves (L), flowers (F), stems (S), and immature fruits (Fr) by ultrasound-assisted extraction (UAE) were assessed for their antioxidant, anti-inflammatory, and growth-inhibitory activity, and chemically characterized by an analytical approach based on liquid chromatography/electrospray/high-resolution tandem–mass spectrometry (LC-ESI/HRMS/MS). The resulting chromatographic profile revealed 30 major constituents belonging to the flavonoids, glucosinolates, phenolic acids, and polar lipids, as well as hydroxy fatty acid classes. Naringenin-di-C-glucoside, isorhamnetin-O-deoxyhexosyl-hexoside, kaempferol-O-dideoxyhexosyl-hexoside, and isorhamnetin O-dideoxyhexoside are reported here for the first time in the genus Reseda. The Fr extract exhibited the highest anti-inflammatory and radical scavenging properties, likely due to its higher flavonoid content compared to the other extracts. On the other hand, the F extract significantly reduced the viability of colorectal adenocarcinoma (CaCo-2) and hepatocarcinoma (HepG-2) cells. Lactate dehydrogenase (LDH) release assay showed that the treatments with R. alba did not induce the release of the marker enzyme in CaCo-2 and HepG-2 cells, suggesting the involvement of a different cell death pathway. Overall, the bioactivities observed among the different plant organs highlight the beneficial potential of R. alba and provide a rationale for future bioactivity-guided isolation studies. Full article

Background: Acute myocardial infarction (AMI) remains the most lethal critical emergency worldwide. Although Angong Niuhuang Pill (ANP) is an established rescue medicine that has demonstrated outstanding therapeutic potential for cardiovascular diseases, its modern molecular mechanism has never been systematically elucidated because of its chemical complexity and unidentified targets. Methods: This study utilizes a multi-layer analytical pipeline of AI mining, network pharmacology, transcriptomics, and experimental confirmation. The components of ANP were comprehensively identified by UHPLC-Q Exactive Orbitrap HRMS. The TranSiGen algorithm was utilized to deeply mine the data and rank the components according to their relevance to AMI. The top 20 components were selected as prior weights and introduced into network pharmacology for analysis. Subsequently, a mouse model of AMI was established by ligating the left coronary artery. Cardiac function in the mice was evaluated by echocardiography and serum biochemical indicators. The pathological changes in the heart tissue were assessed by hematoxylin-eosin (H&E) and Masson staining. Cardiac transcriptome sequencing was performed, and pathway enrichment was analyzed by KEGG. The key pathways were verified by qPCR and immunofluorescence, achieving cross-validation between AI prediction and experimental findings. Results: The identification of ANP resulted in the detection of a total of 73 compounds, and the TranSiGen algorithm was employed to prioritize these compounds, yielding a ranked list of the top 20 candidates. Functional evaluation using echocardiography, serum biochemical markers, and histopathological examination demonstrated that ANP significantly ameliorated cardiac function in mice following myocardial infarction. Integration of network pharmacology and transcriptomic enrichment identified convergent axes of IL-17 signaling and mitochondrial quality control, which were subsequently experimentally validated as mechanisms by which ANP ameliorated cardiac injury. Experimental validation confirmed that ANP downregulated protein expression of IL-17A and TNF-α, normalized PINK1 and LC3-II/LC3-I marker profiles, with concomitant p62 reduction, thereby providing comprehensive molecular evidence at both transcriptional and translational levels to support the AI-driven predictions. Conclusions: This study identified IL-17 signaling and mitochondrial quality control as pathway axes associated with ANP-mediated cardioprotection against AMI, supported by AI-driven compound screening, transcriptome-network cross-validation, and experimental confirmation. This analytical framework may be adaptable to other complex TCM formulas for mechanism exploration and clinical translation. Full article

Lumpy Skin Disease (LSD) is an economically significant viral disease of cattle, widely prevalent across Africa, particularly in sub-Saharan regions. In 2024, Tunisia reported its first outbreak. Understanding the genetic characteristics of lumpy skin disease virus (LSDV) and related poxviruses is critical for surveillance and control. Twenty-nine samples from 26 suspected cases were screened for LSDV using qPCR, followed by a High-Resolution Multiplex Melting (HRM) assay. Three representative samples, two LSDV-positive and one bovine papular stomatitis virus (BPSV)-positive, were subjected to whole-genome sequencing using Pacific Biosciences (PacBio) HiFi long-read technology. Phylogenetic analyses of the LSDV-marker gene RPO30 and complete genomes were performed alongside SNP and InDel profiling. The Tunisian LSDV isolates clustered with Clade 1.2.2 field strains and were 100% identical to each other and to the Italian isolate LSDV_Italy_Sardinia_2025, sharing 99.99% nucleotide identity with LSDV_V281_Nigeria. Although only two LSDV isolates were sequenced which showed no genetic differences, these findings suggest genomic stability within Clade 1.2.2. The Tunisian BPSV isolate showed high similarity (98.15–98.59%) to strains reported in Germany and Switzerland. This study presents the first genetic characterization of LSDV and BPSV in Tunisia, highlighting the importance of accurate differential diagnosis among poxviruses and continuous genomic surveillance to inform control strategies. Full article

The micro-immunotherapy medicine (MIM) 2LEID-N^® was developed to sustain immune response, notably in the framework of respiratory infections. This pilot study investigated the potential effects of one capsule of this MIM, referred to as 2LEID-N-9 throughout the manuscript, containing IL-12 (5 CH), IFN-γ (6 CH), and TNF-α (5 CH). Phagocytosis and surface marker expression were assessed using flow cytometry, and cytokine secretion was assessed by ELISA. Cellular models included human monocyte-derived macrophages, peripheral blood mononuclear cells (PBMCs) from healthy donors, and THP-1 cells. Liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS/MS) was used to detect the actives. Compared with vehicle control, 2LEID-N-9 showed a trend towards the enhanced phagocytic activity of macrophages. In PBMCs, 2LEID-N-9 upregulated the secretion of several cytokines, including IL-2, IL-4, IL-13, IFN-γ, and TNF-α in both basal and CD3/CD28-stimulated conditions. Notably, a tendency towards increased secretion of TNF-α was found in LPS-stimulated THP-1 cells. The presence of the three actives, as assessed by LC-HRMS/MS, combined with the functional data, provide promising exploratory evidence of immunomodulatory effects and tendencies towards the stimulation of innate and adaptive immune cells, warranting further investigation. Full article

Background/Objectives: WenTong HuoXue Cream (WTHXC) plays a significant role in the treatment of diabetic peripheral neuropathy (DPN). However, the material basis and quality control methods for this formulation remain largely unexplored. Methods: In this study, UPLC-HRMS/MS combined with standard reference substances was employed to comprehensively identify and confirm the chemical constituents of WTHXC. Furthermore, a rapid and sensitive UPLC-MS/MS method based on multiple reaction monitoring (MRM) mode was developed and validated for the simultaneous quantification of the marker components. Results: Nine compounds were unambiguous characterized, including Di-hydrocapsaicin (DHC), Oxypeucedanin hydrate (OPH), Imperatorin (IMP), Isoimperatorin (IIMP), Xanthotoxin (XAN), Hydroxysafflor yellow A (HSYA), Chlorogenic acid (CGA), Ferulic Acid (FA) and Ligustilide (LIG). The results of method validation denotes that all the analytes showed good linearity between concentration and peak area in the tested ranges, with correlation coefficients (r) not less than 0.9990. The relative standard deviation (RSD) of precision was in the range of 0.57–7.11%. The accuracy of the method, verified by recovery experiments at three concentration levels, ranged from 96.51% to 101.04% for all analytes. Transdermal behavior determination results demonstrate that OPH, HYSA, CGA, FA and LIG exhibited favorable skin permeability and may serve as the key active components of WTHXC. Conclusions: This study elucidates the material basis of WTHXC, providing a scientific foundation for the development of quality control methods and facilitating its broader clinical application. Full article

Glaucoma remains a primary cause of blindness, yet its pathogenesis often extends beyond intraocular pressure (IOP). This review integrates four converging lines of metabolic evidence—aqueous humor (AH) metabolomics, kynurenine pathway (KP) activity, tetrahydrobiopterin (H4BIP) biology, and NAD/one-carbon dysfunction—into a testable framework for retinal ganglion cell vulnerability. By utilizing a systematic AH metabolomics atlas covering glaucoma, pseudoexfoliation, and diabetes on a standardized HILIC-LC-HRMS platform, we demonstrate that, while aromatic amino acid elevations are non-specific markers, kynurenine monooxygenase (KMO) upregulation is a condition-specific glaucoma signature. These local findings are corroborated by systemic evidence: POAG patients exhibit significant folic acid deficiency (p = 0.007) and elevated alpha-1-antitrypsin (AAT). Critically, AAT correlates inversely with both serum folate (r_s = −0.485, p < 0.001) and retinal nerve fiber layer thickness (r_s = −0.386, p = 0.017), providing the first in-patient evidence linking systemic inflammation to structural optic nerve damage. We conclude that KMO serves as a critical enzymatic node linking tryptophan metabolism, H4BIP availability, and NAD synthesis. These results characterize glaucoma as a disease of progressive cofactor failure and define a research agenda for multimodal metabolic neuroprotection. Full article

Purpose: The purpose of this study was to characterize the distribution and longitudinal evolution of intraretinal and subretinal hyperreflective foci (HF) in treatment-naive neovascular age-related macular degeneration (nAMD), and to examine associations between HF burden, hyperreflective material boundary remodeling (HRM-BR), and best-corrected visual acuity (BCVA) outcomes following bevacizumab treat-and-extend therapy. Methods: This was a retrospective observational study of 84 treatment-naive nAMD eyes receiving intravitreal bevacizumab via a treat-and-extend protocol. Spectral-domain OCT (Revo FC, Optopol) was performed at baseline (M0), month 3 (M3), and month 6 (M6). HF were quantified in the intraretinal and subretinal compartments using ImageJ software (version 1.54, National Institutes of Health, Bethesda, MD, USA) by two masked graders, with inter-rater agreement assessed by intraclass correlation coefficient (ICC). Eyes were classified into four HRM evolution patterns following the framework of Yu et al. Primary outcome was BCVA change from M0 to M6. Multivariable linear regression was performed to assess independent predictors of BCVA change. Results: Baseline intraretinal HF counts increased significantly across HRM Patterns 1 through 4 (median 0, 6, 4, and 8, respectively; Kruskal–Wallis p < 0.001; 95% CI for Spearman r = 0.471: [0.286, 0.623]). A higher baseline intraretinal HF count correlated with worse BCVA change at M6 (r = −0.300, 95% CI [−0.483, −0.092], p_adj = 0.010). In the primary multivariable model (n = 67), both intraretinal HF burden (β = −0.449, 95% CI [−0.879, −0.020], p = 0.041) and HRM width (β = −0.003, 95% CI [−0.005, −0.001], p = 0.014) were independent predictors of BCVA change. The transient M3 intraretinal HF peak in Pattern 3 eyes (median 4 → 12 → 4) was statistically confirmed by Wilcoxon signed-rank testing (M0 → M3: p = 0.004; M3 → M6: p = 0.001). Conclusions: Intraretinal HF burden is a graded marker of HRM pattern severity and an independent predictor of visual outcomes in nAMD, alongside HRM width. The statistically validated transient M3 HF peak in Pattern 3 may represent an early OCT signal of active boundary remodeling. Full article

Chronic kidney disease (CKD) affects more than 10% of the population and is associated with a persistent, low-grade inflammatory state that accelerates tubulointerstitial fibrosis, worsens prognosis, and increases cardiovascular risk. This review aims to synthesize current knowledge on specialized pro-resolving mediators (SPMs) in the context of CKD pathophysiology, biomarkers, and therapeutic potential. We discuss key anti-inflammatory and pro-resolving mechanisms of SPMs that translate into nephroprotective and antifibrotic effects in experimental kidney models. The review summarizes data on EPA/DHA supplementation, including its impact on lipid profiles, inflammatory biomarkers (CRP, IL-6, TNF-α), and oxidative stress in patients with CKD. We also highlight contemporary analytical methods for biomarker assessment (LC-MS/MS, UHPLC-HRMS) and their potential for monitoring inflammatory activity across its phases (initiation, attenuation, resolution), CKD progression, and responses to ω-3/SPM-based interventions. Finally, we discuss the therapeutic potential of SPMs, as well as safety considerations and pharmacological interactions. In conclusion, SPMs and ω-3-derived mediators represent promising research and clinical targets as markers and modulators of inflammation in CKD, but require further validation in well-designed prospective studies. Full article

Ulcerative colitis (UC) is a chronic inflammatory bowel disease that seriously jeopardizes health. Saposhnikovia divaricata (FF) has anti-inflammatory and antioxidant pharmacological effects. Numerous studies have demonstrated the efficacy of FF in alleviating UC, but the potential mechanism by which inhibiting ferroptosis alleviates UC remains unclear. This research aims to investigate the ways in which FF regulates inflammation, oxidative stress and ferroptosis to attenuate UC. Firstly, the chemical compounds of FF were identified by UPLC-Q-Orbitrap HRMS. FF significantly reduced levels of nitric oxide (NO), proinflammatory cytokines and reactive oxygen species (ROS) in the lipopolysaccharide (LPS)-induced inflammation in RAW264.7 macrophage cells. We used 3% Dextrose sulfate sodium (DSS) to establish the UC model in C57BL/6 mice. FF (8.4 g/kg) effectively ameliorated the symptoms of weight loss and colon damage, significantly attenuating oxidative stress and modulated the levels of ferroptosis markers in the colon. Moreover, FF can down-regulate the expression of p53 protein and up-regulate the expression of SLC7A11 and GPX4 proteins. The results showed that FF can inhibit inflammation, oxidative stress, and ferroptosis to alleviate DSS-induced UC, by regulating the p53 signaling pathway. Full article

Mushroom-related intoxications pose a distinctive challenge for forensic medicine because early manifestations are non-specific, latency may be prolonged, and co-exposures can obscure the mechanism of death. This narrative review summarizes key toxic and psychoactive fungi and their principal compounds, spanning organ-toxic syndromes (amatoxins, orellanine) and functional neuropsychiatric intoxications—acute, predominantly functional effects causing impairment rather than organ failure (psilocybin/psilocin, ibotenic acid/muscimol). We propose an integrated diagnostic workflow combining exposure history, biochemical markers of organ injury, mycological assessment, and confirmatory toxicology. Particular emphasis is placed on postmortem interpretation: toxin instability and biotransformation, conjugation, matrix effects, postmortem redistribution (central vs. femoral blood), and postmortem fungal colonization that may alter analyte profiles or generate misleading metabolites. Because robust lethality thresholds are unavailable for most mushroom toxins, conclusions should rely on a multi-source synthesis of scene information, autopsy/histopathology, and time-dependent matrix selection (urine, gastric contents/vomitus, bile, and selected tissues; kidney for late orellanine confirmation). We review current screening and confirmatory methods—ELISA; LC-MS/MS, LC-HRMS/MS, GC-MS—and highlight pre-analytical requirements (rapid sampling, cold storage) to reduce false negatives. Finally, we discuss emerging directions such as point-of-care tests, portable mass spectrometry, and DNA barcoding for species identification. Full article

Background/Objectives: Anatolia hosts a rich diversity of Hypericum taxa; however, the chemical and biological properties of most species remain insufficiently characterized. Methods: This study combined liquid chromatography–high-resolution mass spectrometry (LC–HRMS) with high-performance liquid chromatography coupled to diode-array detection (HPLC–DAD) to profile 14 extracts obtained from 12 Anatolian species together with H. perforatum, and to examine whether metabolic variation aligns with bioactivity trends. Results: Chemometric analyses (Principal Component Analysis—PCA—and Hierarchical Cluster Analysis—HCA) revealed distinct chemotypes primarily driven by phenolic acids and flavonol glycosides, with hyperoside emerging as a consistently abundant marker. Phenolic-rich extracts displayed enhanced functional properties in multiple assays. Among them, H. triquetrifolium showed the strongest acetylcholinesterase inhibition and notable antioxidant capacity, whereas the selected taxa exhibited comparatively high tyrosinase or $\alpha$-glucosidase inhibition. Antimicrobial testing indicated narrow-spectrum activity limited to Gram-positive bacteria and Candida spp. Conclusions: The integration of metabolite profiling with chemometric and functional data highlights clear chemotype–bioactivity associations across Anatolian Hypericum taxa and underscores the analytical value of LC–HRMS-guided chemotyping for pharmacognostic assessments. These results broaden the phytochemical and bioactivity landscape of this genus and provide a comparative framework for prioritizing species for future targeted investigations. Full article

Natural compounds are increasingly explored for their ability to modulate multiple molecular pathways involved in inflammation and oxidative stress and for their therapeutic potential. Among these, Opuntia ficus-indica (L.) Mill. has attracted growing interest due to its rich phytochemical profile; however, the biological properties of unripe fruits remain largely unexplored. In this study, a hydroalcoholic extract obtained from unripe O. ficus-indica fruits was characterized for its chemical composition, antioxidant capacity, and concentration-dependent embryotoxic profile and subsequently investigated in a zebrafish model of spinal cord injury (SCI). UHPLC-HRMS/MS analysis identified 14 secondary metabolites, mainly flavonoids and phenylpropanoid acids. Antioxidant activity was confirmed by DPPH and ABTS assays. An embryotoxicity assessment conducted according to OECD Test Guideline 236 revealed no mortality at concentrations below 100 µg mL⁻¹ and an LC₅₀ of 323.59 µg mL⁻¹ at 96 h post-fertilization, allowing the identification of non-toxic concentrations for subsequent in vivo experiments. Based on these results, the extract was tested in a larval zebrafish SCI transection model. Treated larvae showed improved locomotor recovery, particularly under continuous exposure, accompanied by modulation of molecular pathways involved in inflammation, neurotrophic support, and neurogenesis, including reduced pro-inflammatory cytokine expression and increased BDNF and Sonic Hedgehog signaling markers. Overall, these findings expand current knowledge on unripe O. ficus-indica and highlight its potential to modulate molecular pathways involved in SCI-induced damage and repair. Full article

Background: Tendinopathy remains a prevalent musculoskeletal disorder with limited disease-modifying pharmacotherapy. This study aimed to identify a reparative agent from the traditional medicinal herb Eucommiae Cortex and elucidate its mechanism of action. Methods: A bioactive fraction was first identified through a bioactivity-guided strategy using tenocyte cytoprotection and migration assays, then characterized by UHPLC-HRMS/MS. Its major constituent, aucubin (AU), which mirrors the fraction’s key pharmacological activities, was evaluated both in vitro and in vivo. In H₂O₂-injured tenocytes, AU’s effects on viability, apoptosis, oxidative stress (ROS, MDA, SOD) and inflammation (IL-1β, TNF-α) were assessed, with specific focus on estrogen receptor (ER) pathway involvement using pharmacological tools (17β-estradiol and (R, R)-THC). In a collagenase-induced Achilles tendinopathy model using male SD rats, AU’s therapeutic efficacy was evaluated via multimodal assessment: ultrasonography, histopathology (H&E, Masson’s trichrome, Sirius red), TEM, immunohistochemistry, and biochemical analysis of tissue markers. Results: AU effectively attenuated H₂O₂-induced tenocyte injury by enhancing viability, reducing apoptosis, and mitigating oxidative/inflammatory stress. These effects were mimicked by 17β-estradiol and reversed by the selective ERβ antagonist (R, R)-THC, indicating ERβ dependence. In vivo, AU treatment promoted structural and functional recovery, improved collagen maturity (increased Col I/Col III ratio and fibril diameter), suppressed matrix degradation (MMP-3, MMP-13) and apoptosis, and reduced oxidative stress and inflammation in tendon tissue. Conclusions: This study identifies aucubin as a novel phytoestrogenic compound from Eucommiae Cortex that promotes tendon repair through an ERβ-mediated mechanism. These findings position ERβ activation as a promising therapeutic strategy for tendinopathy and highlight AU as a promising lead compound for further development. Full article

This study investigated single-nucleotide polymorphisms (SNPs) within the CAPN1 316, CAPN1 4751, and CAST 2959 markers using high-resolution melting (HRM) analysis to predict meat tenderness in crossbred beef cattle. Tenderness was assessed using the Warner–Bratzler shear force (WBSF) test, with results expressed in grams (g), representing the force required to shear muscle fibers. Significant differences in phenotypic data were observed among the genotypic groups. The finding showed that polymorphisms at CAPN1 316, CAPN1 4751, and CAST 2959 exert interactive effects on meat quality traits. Notably, the TT genotype at CAPN1 4751 increased the adjusted WBSF (aWBSF) by approximately 792 g, indicating that TT was an unfavorable variant for tenderness. These results support the use of marker-assisted selection strategies in which the TT genotype is managed to minimize its frequency while other relevant markers are concurrently monitored, thereby enhancing genetic progress in meat tenderness across commercial cattle populations. This study demonstrated that CAPN1 4751 could serve as an effective marker for genetic selection in crossbred beef cattle and confirmed the efficiency of HRM analysis as a molecular tool for SNP genotyping. In conclusion, the findings provided an alternative approach for SNP detection in livestock breeding programs and represented an important step toward improving meat quality, meeting consumer expectations, and supporting the long-term sustainability of Thailand’s beef industry. The results highlighted the polygenic nature of meat tenderness and emphasized the importance of integrating multiple SNP markers to accurately assess the genetic potential for meat quality traits in cattle. Full article