Search Results (123)

Background: Acute mountain sickness (AMS) is a prevalent and potentially life-threatening condition caused by rapid exposure to high-altitude hypoxia, affecting pulmonary and neurological functions. Tongqiao Jiuxin Oil (TQ), a traditional Chinese medicine formula composed of aromatic and resinous ingredients such as sandalwood, agarwood, frankincense, borneol, and musk, has been widely used in the treatment of cardiovascular and cerebrovascular disorders. Clinical observations suggest its potential efficacy against AMS, yet its pharmacological mechanisms remain poorly understood. Methods: The chemical profile of TQ was characterized using UHPLC-Q-Exactive Orbitrap HRMS. Network pharmacology was applied to predict the potential targets and pathways involved in AMS. A rat model of AMS was established by exposing animals to hypobaric hypoxia (~10% oxygen), simulating an altitude of approximately 5500 m. TQ was administered at varying doses. Physiological indices, oxidative stress markers (MDA, SOD, GSH), histopathological changes, and the expression of hypoxia- and apoptosis-related proteins (HIF-1α, VEGFA, EPO, Bax, Bcl-2, Caspase-3) in lung and brain tissues were assessed. Results: A total of 774 chemical constituents were identified from TQ. Network pharmacology predicted the involvement of multiple targets and pathways. TQ significantly improved arterial oxygenation and reduced histopathological damage in both lung and brain tissues. It enhanced antioxidant activity by elevating SOD and GSH levels and reducing MDA content. Mechanistically, TQ downregulated the expression of HIF-1α, VEGFA, EPO, and pro-apoptotic markers (Bax/Bcl-2 ratio, Caspase-3), while upregulated Bcl-2, the anti-apoptotic protein expression. Conclusions: TQ exerts protective effects against AMS-induced tissue injury by improving oxygen homeostasis, alleviating oxidative stress, and modulating hypoxia-related and apoptotic signaling pathways. This study provides pharmacological evidence supporting the potential of TQ as a promising candidate for AMS intervention, as well as the modern research method for multi-component traditional Chinese medicine. Full article

Background/Objectives: Senecio scandens Buch.-Ham. is a flavonoid-rich traditional medicinal plant with established immunomodulatory properties. However, the mechanisms underlying the immunoregulatory and intestinal protective effects of its flavonoid extract (Senecio scandens flavonoids—SSF) remain unclear. This study characterized SSF’s bioactive components and evaluated its efficacy against cyclophosphamide (CTX)-induced immunosuppression and intestinal injury. Methods: The constituents of SSF were identified using UHPLC/Q-Orbitrap/HRMS. Mice with CTX-induced immunosuppression were treated with SSF (80, 160, 320 mg/kg) for seven days. Immune parameters (organ indices, lymphocyte proliferation, cytokine, and immunoglobulin levels) and gut barrier integrity markers (ZO-1, Occludin, Claudin-1 protein expression; sIgA secretion; microbiota composition) were assessed. Network pharmacology combined with functional assays elucidated the underlying regulatory mechanisms. Results: Twenty flavonoids were identified in SSF, with six prototype compounds detectable in the blood. The SSF treatment significantly ameliorated CTX-induced weight loss and atrophy of the thymus and spleen. It enhanced splenic T- and B-lymphocyte proliferation by 43.6% and 29.7%, respectively; normalized the CD4⁺/CD8⁺ ratio (1.57-fold increase); and elevated levels of IL-2, IL-6, IL-10, TNF-α, IFN-γ, IgM, and IgG. Moreover, SSF reinforced the intestinal barrier by upregulating tight junction protein expression and sIgA levels while modulating the gut microbiota, enriching beneficial taxa (e.g., the Lachnospiraceae_NK4A136_group, Akkermansia) and suppressing pathogenic Alistipes. Mechanistically, SSF activated the TLR/MyD88/NF-κB pathway, with isoquercitrin identified as a pivotal bioactive constituent. Conclusions: SSF effectively mitigates CTX-induced immunosuppression and intestinal damage. These findings highlight SSF’s potential as a dual-functional natural agent for immunomodulation and intestinal protection. Subsequent research should validate isoquercitrin’s molecular targets and assess SSF’s clinical efficacy. Full article

The current study evaluated the anticonvulsant properties of ethanolic extracts from Morus alba, Angelica archangelica, Passiflora incarnata, and Valeriana officinalis using integrated phytochemical, in vivo, biochemical, and computational approaches. Phytochemical analysis by UHPLC-HRMS/MS revealed the presence of various bioactive compounds, notably flavonoids such as isorhamnetin, quercetin, and kaempferol. In an electroshock-induced seizure model, Morus alba extract (MAE, 100 mg/kg) demonstrated significant anticonvulsant effects, reducing both seizure duration and incidence, likely mediated by flavonoid interactions with GABA-A and 5-HT3A receptors, as suggested by target prediction and molecular docking analyses. The extracts of Angelica archangelica (AAE, 100 mg/kg) and Passiflora incarnata (PIE, 50 mg/kg) exhibited moderate, non-significant anticonvulsant activities. At the same time, Valeriana officinalis (VOE, 50 mg/kg) displayed considerable antioxidant and anti-inflammatory properties but limited seizure protection. All extracts significantly reduced brain inflammation markers (TNF-α) and enhanced antioxidant defenses, as indicated by total thiols. Molecular docking further supported the interaction of key phytochemicals, including naringenin and chlorogenic acid, with human and mouse 5-HT3A receptors. Overall, Morus alba extract exhibited promising therapeutic potential for epilepsy management, warranting further investigation into chronic seizure models and optimized dosing strategies. Full article

Aletris spicata (Thunb.) Franch. (AS), a traditional edible and medicinal plant for treating asthma, was investigated for its therapeutic mechanisms. Liquid chromatography-high-resolution tandem mass spectrometry (LC-HRMS/MS) analysis identified 33 compounds in AS. In ovalbumin (OVA)-induced asthmatic mice, AS significantly reduced inflammatory cells (neutrophils, lymphocytes, eosinophils) in bronchoalveolar lavage fluid (BALF) and decreased IL-4, IL-5, IL-13, TNF-α, and serum IgE while increasing IFN-γ. AS alleviated lung and intestinal inflammation, reduced ROS and MDA levels, and enhanced SOD activity. Immunohistochemistry and Western blot revealed AS upregulated Nrf2/HO-1 expression and inhibited NF-κB p65 nuclear translocation. Gut microbiota studies demonstrated AS restored intestinal flora homeostasis by modulating the richness, diversity, and composition. Spearman correlation analysis identified significant relationships between oxidative stress markers, inflammatory cytokines, and specific gut bacteria. These findings indicate that AS mitigates asthma through antioxidant effects (Nrf2/HO-1 pathway), anti-inflammatory actions (NF-κB pathway), and gut microbiota modulation. The study provides a scientific basis for developing AS as a natural anti-asthma treatment or functional food. The multi-target mechanism involving oxidative stress, inflammation, and gut flora highlights AS’s comprehensive therapeutic potential for asthma management. Full article

Nitazenes represent an emerging class of new synthetic opioids characterized by a high-potency μ-opioid receptor (MOR) agonist activity. Background: We report two 20-year-old males who presented with severe neurorespiratory depression with typical opioid syndrome, but no opioid identification based on routine blood and urine screening tests. The first patient recovered with supportive care, mechanical ventilation, and naloxone infusion, whereas the second patient developed post-anoxic cardiac arrest and died from brain death. Methods: A complementary comprehensive toxicological screening using liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS) was performed, and data were processed using a dedicated molecular network strategy to profile the metabolites. Results: Protonitazene and protonitazepyne, two nitazenes differing in their ethylamine moieties (i.e., a diethyl versus a pyrrolidine substitution, respectively), were identified. We found an extensive metabolism of protonitazene, leading to the identification of multiple phase I (resulting from hydroxylation, N-desethylation, and O-despropylation) and phase II (resulting from glucuronidation) metabolites. By contrast, protonitazepyne metabolism appeared limited, with one metabolite annotated confidently, protonitazepyne acid, which resulted from the oxidative pyrrolidine ring cleavage. Concusions: To conclude, nitazene detection is highly challenging due to its extensive structural and metabolic diversity. Our findings highlight the contribution of the untargeted LC-HRMS screening approach and suggest that diagnostic product ions can serve as robust markers for nitazene identification. Full article

Many studies have shown that only a few breeds of goat have the trait of high fertility, and genes play an important role in regulating litter size. This study investigated the effects of GnRHR, BMP6, and FSHR gene polymorphisms on litter traits in four goat breeds (Chongqing black, Chuanzhong black, Leizhou, and Nubian). Single nucleotide polymorphisms (SNPs) (g.75G > A in GnRHR, g.951T > C in BMP6, and g.-112C > T/g.3236C > A in FSHR) were genotyped using PCR-RFLP/HRM in 959 goats. Association analysis revealed significant correlations between these SNPs and litter size in the Chongqing, Chuanzhong, and Leizhou breeds, with genotypes AA (GnRHR), CC (BMP6), TT (FSHR), and AA (FSHR) linked to higher prolificacy. Polygene polymerizing effect analysis identified the optimal combinations (e.g., AATTCC, AACCAACC) with enhanced litter sizes. Tissue-specific qPCR in Chuanzhong goats showed GnRHR, BMP6, and FSHR were significantly more highly expressed in reproductive tissues (pituitary, breast, ovary, oviduct) of the prolific group than those in the non-prolific group. These SNPs serve as potential molecular markers for improving goat litter traits through polygenic selection, emphasizing the synergistic impact of multi-gene interactions on prolificacy. Full article

A total of 31 compounds were isolated from the ethyl acetate and n-butanol fractions of Anchusa italica Retz., which contained one ursane triterpenoid, 2α,3β,19α-trihydroxy-23-formyl-urs-12-en-28,21β-olide (1), and five norisoprenoids: (2R,6R,9S)-9-hydroxy-4-megastigmen-3-one-2-O-β-D-glucopyranoside (3); (2R,6S,9S)-9-hydroxy-megastigman-4,7-dien-3-one-2-O-β-D-glucopyranoside (4); (+)-isololiolide β-D-glucopyranoside (5); (2S,8R)-loliolide β-D-glucopyranoside (6a); and (2R,8S)-loliolide β-D-glucopyranoside (6b). It also contained 25 known compounds (2 and 7–30). The chemical structures of the compounds, inclusive of their absolute configurations, were ascertained using spectroscopic methods such as NMR, HR-MS, and quantum chemical calculations (computational NMR and ECD), in combination with relevant literature data. Moreover, the chemotaxonomic significance of the isolated substances was discussed, with compounds 1, 2, and 7–13 potentially broadening the application of triterpenes as taxonomic markers for the classification of the genus Anchusa. Full article

Introduction: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) underwent significant mutations, resulting in the Omicron variant. Methods: In this study, we analyzed blood samples from 98 patients with acute coronavirus disease 19 (COVID-19) hospitalized during the initial SARS-CoV-2 wave and the onset of Omicron in 2021. High-resolution melting (HRM) analysis of PCR products was used to analyze RNA extracted from clinical samples collected in July and November 2021 from patients infected with SARS-CoV-2. Results: HRM analysis revealed a characteristic deletion in the N protein RNA of the virus isolated in November 2021, associated with the Omicron variant. Elevated levels of inflammatory markers and interleukin-6 (IL-6) were observed in both waves of COVID-19. Complement levels and IgG and IgM antibodies to SARS-CoV-2 were detected more often during the second wave. An increase in hemagglutinin-inhibiting (HI) antibodies against influenza viruses was observed in paired blood specimens from moderate to severe COVID-19 patients during both outbreaks. Conclusions: Patients admitted during both waves of COVID-19 showed a significant rise in inflammatory markers, suggesting that Omicron triggers inflammatory responses. The rapid formation of IgM and IgG in Omicron may indicate a faster immune response. Seasonal flu may negatively impact the clinical course of coronavirus infections. Full article

The mutation in NF2 is the most common alteration associated with meningioma oncogenesis, and it is related to the loss of a suppressing protein called merlin. At the same time, alterations in energy production are visible in cancer cells, where increased demands for energy are observed. Fatty acid oxidation could be one of the ways cancer cells obtain energy. This metabolic pathway uses the acylcarnitine shuttle system, which is responsible for the acylation of fatty acids and their transport through the mitochondria. Therefore, this study aimed to profile acylcarnitines with short, medium, and long acyl chain lengths in meningiomas to assess their changes in tumors with different NF2 mutation statuses. For the analysis, solid-phase microextraction (SPME) coupled with liquid chromatography–high-resolution mass spectrometry (LC-HRMS) was used. The presented sampling method enabled less invasive and easy collection of the analytes from the studied lesions, which can be crucial for future analysis of potential biomarkers in the surgery room. It was observed that higher levels of these analytes characterized meningiomas with NF2 mutations. Moreover, the increased energy consumption and elevated levels of acylcarnitines show that these analytes can be considered markers of increased fatty acid oxidation in NF2 mutated cells. Full article

Honey, a sweet and nutritious food produced by honeybees, is extensively consumed by humans due to its potential health benefits. Unfortunately, the adulteration of honey with inexpensive sugar syrups is a prevalent issue. Verifying the authenticity of honey is crucial for maintaining its quality and safety standards. The aim of this study was to identify the illicit addition of sugar syrups to honey imported into the European Union (EU). The European Commission’s Joint Research Centre (JRC) has employed different analytical approaches to detect several markers of adulteration in honey; however, this paper mainly focuses on the use of Liquid Chromatography–High-Resolution Mass Spectrometry (LC-HRMS). Two qualitative methods were developed to detect mannose (Man), difructose anhydride III (DFA III), 2-acetylfuran-3-glucopyranoside (AFGP), and oligo-/polysaccharides with degrees of polymerization (DPs) of 6 to 11. Out of the 320 honey samples provided by the authorities of the participating EU Member States, 147 (46%) were suspicious for non-compliance with the EU Honey Directive 2001/110/EC, mostly due to the presence of mannose and oligo-/polysaccharides. As a result, the development and standardization of sophisticated and universally recognized testing procedures will increase the capability of official control laboratories to detect honey adulteration and will serve as a powerful preventive measure against fraudulent practices in the global honey market. Full article

Chronic drug treatment can alter the lipidome of cancer cells, potentially leading to significant biological changes, such as drug resistance or increased tumor aggressiveness. This study examines the lipidome profiles of four human colorectal cancer (CRC) cell lines, comparing treatment-naïve cells with the same cells after chronic exposure to a clinically used combination therapy (FOLFOXIRI: folinic acid, 5-fluorouracil, oxaliplatin, and irinotecan). Lipidomic profiling was obtained with untargeted liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS). For data deconvolution and to interpret the multifactorial dataset generated, Analysis of Variance Multiblock Orthogonal Partial Least Squares (AMOPLS) was used. Our results indicate specific shifts in triglycerides (TGs), sphingolipids, and phospholipids in CRC cells resistant to FOLFOXIRI. The overall shift in TGs, phosphatidylcholine, and cholesteryl ester species was notably linked to FOLFOXIRI resistance (-R) in SW620 cells, whereas an increased abundance of phospholipids, mainly hexosylceramide and sphingomyelin, was present in the signatures of HCT116-R, LS174T-R, and DLD1-R cells. These altered lipid species may serve as potential prognostic markers in CRC following chemotherapy. Furthermore, lipid-targeting therapies aimed at reprogramming the lipid profiles of drug-resistant cells could play a crucial role in restoring drug sensitivity and improving patient survival. Full article

Qualitative and quantitative differences in the chemical composition between bee pollen originated from Castanea sativa (Türkiye and Slovenia), Salix spp. (Türkiye and Slovenia), and Quercus spp. (Türkiye) and androecia of Castanea sativa, Salix alba, and Quercus pubescens (apetalous trees) were evaluated for the first time by new high-performance thin-layer chromatography (HPTLC) and ultra-performance liquid chromatography (UPLC) methods using marker compounds. N¹,N⁵,N¹⁰-tricaffeoylspermidine was isolated, and its structure was elucidated by nuclear magnetic resonance (NMR) and high-resolution mass spectrometry (HRMS). It was the main and the marker compound common to bee pollen (≈3–41 mg/g) and androecia (≈3–6 mg/g) samples. To the best of our knowledge, this is the first report of the identification of N¹,N⁵,N¹⁰-tricaffeoylspermidine in bee pollen originated from Salix spp. and androecia of C. sativa, S. alba, and Q. pubescens. The botanical origins of bee pollen were determined via phytochemical profiling using HPTLC-image analyses showing that bee pollen from the same botanical source had almost identical profiles regardless of collection location, geographical differences, and the bee race. In vitro tests and HPTLC-effect-directed analyses (EDAs) were performed to assess antioxidant and xanthine oxidase (XO) inhibitory activities of bee pollen, androecia, and N¹,N⁵,N¹⁰-tricaffeoylspermidine. HPTLC-EDA combined with image analyses was used for comparing the activities of bee pollen, androecia, N¹,N⁵,N¹⁰-tricaffeoylspermidine, and also other marker compounds (quercetin, myricitrin, hyperoside, quercitrin, isoquercitrin, and rutin). The remarkable bioactivity of N¹,N⁵,N¹⁰-tricaffeoylspermidine was for the first time evaluated by HPTLC-EDA and in vitro tests. This is the first study performing HPTLC-XO inhibitory activity analyses on the HPTLC NH₂ F_254S plates. Further bioactivity studies on botanically and chemically well-characterized bee pollen samples are needed to aid in the use of bee pollen-containing supplements in the prevention and treatment of diseases. Full article

Diverse feeding habits in teleosts involve a wide range of appetite-regulating factors. As an appetite-suppressing gene, the polymorphisms of POMCa in largemouth bass (Micropterus salmoides) were validated via sequencing and high-resolution melting (HRM). The frequency distribution of different POMCa genotypes were analyzed in two populations, and physiological responses of different POMCa genotypes to feed domestication were investigated. The indel of an 18 bp AU-rich element (ARE) in the 3′ UTR and four interlocked SNP loci in the ORF of 1828 bp of POMCa cDNA sequence were identified in largemouth bass and constituted three genotypes of POMC-A I, II, and III, respectively. POMC-A I and Allele I had increased frequencies in the selection population than in the non-selection population (p < 0.01), 63.55% vs. 43.33% and 0.7850 vs. 0.6778, respectively. POMC-A I possessed the lowest value of POMCa mRNA during fasting (p < 0.05) and exhibited growth and physiological advantages under food deprivation and refeeding according to the levels of body mass and four physiological indicators, i.e., cortisol (Cor), growth hormone (GH), insulin-like growth factor-1 (IGF-1), and glucose (Glu). The identification of three POMCa genotypes, alongside their varying physiological responses during feed domestication, suggests a selective advantage that could be leveraged in molecular marker-assisted breeding of largemouth bass that are adapted to feeding on formula diet. Full article

Honey differentiation based on the botanical origin is crucial to guarantee product authenticity, especially considering the increasing number of fraud cases. This study assessed the metabolomic differences arising from various botanical origins in honey products sold in Spanish markets, focusing on two goals: (1) discrimination within monofloral samples (eucalyptus, rosemary, and orange blossom honey) and (2) differentiation between multifloral vs. monofloral honey samples. An omics strategy based on ultra-high-performance liquid chromatography coupled with quadrupole-Orbitrap-high-resolution mass spectrometry (UHPLC-Q-Orbitrap-HRMS) was applied for the reliable identification of specific honey markers selected by orthogonal partial least squares discriminant analysis (OPLS-DA) (R²Y = 0.929–0.981 and Q² = 0.868–0.952), followed by the variable importance in projection (VIP) approach. Key amino acid, alkaloid, and trisaccharide markers were identified to distinguish between honey samples. Some Amadori compounds were highlighted as eucalyptus honey markers, suggesting their potential use for honey aging and botanical origin differentiation. L-phenylalanine and raffinose were markers of rosemary honey. Four markers (e.g., trigonelline, L-isoleucine, and N-(1-deoxy-1-fructosyl)isoleucine) were found in higher levels in multifloral samples, indicating a greater availability of amino acids, potentially increasing the Maillard reaction. This research is the first to address the botanical origin’s impact on honey by identifying novel markers not previously described. Full article

The purple carrot cultivar ‘Purple Sun’ (Daucus carota L.) is characterized by a relevant content of phenolic compounds and anthocyanins, which may play an important role in reducing the risk of chronic diseases and in the treatment of metabolic syndrome. In the present study, the genetic diversity, phytochemical composition, and bioactivities of this outstanding variety were studied for the first time. Genetic analysis by molecular markers estimated the level of genetic purity of this carrot cultivar, whose purple-pigmented roots were used for obtaining the purple carrot ethanol extract (PCE). With the aim to identify specialized metabolites potentially responsible for the bioactivities, the analysis of the metabolite profile of PCE by LC-ESI/LTQ Orbitrap/MS/MS was carried out. LC-ESI/HRMS analysis allowed the assignment of twenty-eight compounds, putatively identified as isocitric acid (1), phenolic acid derivatives (2 and 6), hydroxycinnamic acid derivatives (9, 10, 12–14, 16, 17, 19, 22, and 23), anthocyanins (3–5, 7, 8, 11, and 18), flavanonols (15 and 21), flavonols (20 and 24), oxylipins (25, 26, and 28), and the sesquiterpene 11-acetyloxytorilolone (27); compound 26, corresponding to the primary metabolite trihydroxyoctanoic acid (TriHOME), was the most abundant compound in the LC-ESI/HRMS analysis of the PCE, and hydroxycinnamic acid derivatives followed by anthocyanins were the two most represented groups. The antioxidant activity of PCE, expressed in terms of reactive oxygen species (ROS) level and antioxidant enzymes activity, and its pro-metabolic effect were evaluated. Moreover, the antibacterial activity on Gram (−) and (+) bacterial strains was investigated. An increase in the activity of antioxidant enzymes (SOD, CAT, and GPx), reaching a maximum at 0.5 mg/mL of PCE with a plateau at higher PCE concentrations (1.25, 2.5, and 5.0 mg/mL), was observed. PCE induced an initial decrease in ROS levels at 0.1 and 0.25 mg/mL concentrations, reaching the ROS levels of control at 0.5 mg/mL of PCE with a plateau at higher PCE concentrations (1.25, 2.5, and 5.0 mg/mL). Moreover, significant antioxidant and pro-metabolic effects of PCE on myoblasts were shown by a reduction in ROS content and an increase in ATP production linked to the promotion of mitochondrial respiration. Finally, the bacteriostatic activity of PCE was shown on the different bacterial strains tested, while the bactericidal action of PCE was exclusively observed against the Gram (+) Staphylococcus aureus. The bioactivities of PCE were also investigated from cellular and molecular points of view in colon and hematological cancer cells. The results showed that PCE induces proliferative arrest and modulates the expression of important cell-cycle regulators. For all these health-promoting effects, also supported by initial computational predictions, ‘Purple Sun’ is a promising functional food and an optimal candidate for pharmaceutical and/or nutraceutical preparations. Full article