Search Results (182)

In this study, the effects of water bath and ultrasonic bath systems on bioactive properties, phenolic components and fatty acid profiles of unroasted and roasted pumpkin seeds were investigated. It is thought that determining the bioactive components, phenolic constituents and fatty acid profiles of unroasted and roasted pumpkin seeds will lead to the establishment of usage norms according to their composition characteristics. Total phenolic quantities of the pumpkin seed extracts obtained by water bath extraction of the seeds were defined to be between 7.58 (control) and 11.55 (25 min) and 10.20 (control) and 17.18 mg GAE/100 g (50 min), respectively. Phenolic content increased by 50% after 50 min of ultrasonic extraction, indicating the efficiency of this method. Also, total flavonoid amounts increased about 55% after 25 min of ultrasonic extraction, indicating the efficiency of this method. It was observed that the catechin contents of unroasted pumpkin seeds obtained in water and ultrasonic baths decreased significantly at the 50th minute of extraction compared to the control. The antioxidant activity values (DPPH) of roasted pumpkin seeds treated in water- and ultrasonic bath systems increased by approximately 10% compared to the control at 50 min of sonication in both systems, respectively. Also, the 3,4-dihydroxybenzoic acid amounts of the extracts obtained by both extraction systems of roasted pumpkin seeds were determined between 9.85 (50 min) and 17.22 mg/100 g (control) and 11.17 (25 min) and 13.74 mg/100 g (50 min), respectively. The linoleic acid amounts of unroasted pumpkin seed oils extracted in water- and ultrasonic baths varied between 52.34 (50 min) and 53.33% (control) to 52.90 (50 min) and 53.04% (control), respectively. The linoleic acid values of the roasted pumpkin seed oils were established to be between 52.30 (50 min) and 52.84 (25 min) and 52.32 (50 min) and 53.46% (25 min), respectively. In general, the phenolic compound amounts of roasted pumpkin seeds were higher than those of unroasted ones. The fatty acid amounts of pumpkin seed oils extracted with an ultrasonic bath were generally slightly higher than those extracted with a water bath. In future studies, changes in the phytochemical and bioactive properties of pumpkin seed oils obtained by applying different roasting techniques and extraction methods will be investigated. Full article

Background/Objectives: In recent years, lipids have emerged as critical regulators of different disease processes, being involved in cancer pathogenesis, progression, and outcome. Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Imaging (MALDI-MSI) has significantly expanded the technology’s reach, enabling spatially resolved profiling of lipids directly from tissue, including formalin-fixed paraffin-embedded (FFPE) specimens. In this context, MALDI matrix selection is crucial for lipid extraction and ionization, influencing key aspects such as molecular coverage and sensitivity, especially in such specimens with already depleted lipid content. Thus, in this work, we aim to explore the feasibility of mapping lipid species in FFPE clinical samples with MALDI-MSI using 6-aza-2-thiothymine (ATT) as a matrix of choice. Methods: To do so, ATT performances were first compared to those two other matrices commonly used for lipidomic analyses, 2′,5′-dihydroxybenzoic acid (DHB) and Norharmane (NOR), on lipid standards. Results: As a proof-of-concept, we then assessed ATT’s performance for the MALDI-MSI analysis of lipids in FFPE brain sections, both in positive and negative ion modes, comparing results with those obtained from other commonly used dual-polarity matrices. In this context, ATT enabled the putative annotation of 98 lipids while maintaining a well-balanced detection of glycerophospholipids (60.2%) and sphingolipids (32.7%) in positive ion mode. It outperformed both DHB and NOR in the identification of glycolipids (3%) and fatty acids (4%). Additionally, ATT exceeded DHB in terms of total lipid count (62 vs. 21) and class diversity and demonstrated performance comparable to NOR in negative ion mode. Moreover, ATT was applied to a FFPE glioblastoma tissue microarray (TMA) evaluating the ability of this matrix to reveal biologically relevant lipid features capable of distinguishing normal brain tissue from glioblastoma regions. Conclusions: Altogether, the results presented in this work suggest that ATT is a suitable matrix for pathology imaging applications, even at higher lateral resolutions of 20 μm, not only for proteomic but also for lipidomic analysis. This could enable the use of the same matrix type for the analysis of both lipids and peptides on the same tissue section, offering a unique strategic advantage for multi-omics studies, while also supporting acquisition in both positive and negative ionization modes. Full article

Wild ales represent a diverse category of spontaneously fermented beers, influenced by complex microbial populations and variable ingredients. This study employed an integrated metabolomic profiling approach combining proton nuclear magnetic resonance (¹H NMR) spectroscopy, liquid chromatography–mass spectrometry (LC-MS), and spectrophotometric assays (DPPH and FRAP) to characterize the molecular composition and antioxidant potential of 22 wild ales from six countries. A total of 53 compounds were identified and quantified using NMR, while 62 compounds were identified by using LC-MS. The compounds in question included organic acids, amino acids, sugars, alcohols, bitter acids, phenolic compounds, and others. Ingredient-based clustering revealed that the addition of dark fruits resulted in a significant increase in the polyphenolic content and antioxidant activity. Concurrently, herb-infused and light-fruit beers exhibited divergent phytochemical profiles. Prolonged aging (>18 months) has been demonstrated to be associated with increased levels of certain amino acids, fermentation-derived aldehydes, and phenolic degradation products. However, the influence of maturation duration on the antioxidant capacity was found to be less significant than that of the type of fruit. Country-specific metabolite trends were revealed, indicating the influence of regional brewing practices on beer composition. Correlation analysis was employed to identify the major contributors to antioxidant activity, with salicylic, dihydroxybenzoic, and 4-hydroxybenzoic acids being identified as the most significant. These findings underscore the biochemical intricacy of wild ales and exemplify metabolomics’ capacity to correlate compositional variation with functionality and authenticity in spontaneously fermented beverages. Full article

Autoimmune hepatitis (AIH) is linked to an increased risk of hepatocellular carcinoma (HCC). However, the precise connection between the two remains unclear. GPR81, a G-protein-coupled receptor located on the membranes of various cell types, plays a role in numerous physiological processes. We established an AIH animal model and activated GPR81 using the agonist 3,5-dihydroxybenzoic acid (3,5-DHBA). Additionally, the effect of GPR81 inhibition on tumor and immune cell dynamics was examined using the HepG2, Hep3B, and Hepa1-6 cell lines with the antagonist 3-hydroxybutyric acid (3-OBA). Our results demonstrated that 3,5-DHBA treatment reduced T cell and pro-inflammatory cytokine secretion, while MDSC secretion increased, inhibiting Concanavalin A (Con A)-induced AIH. The inhibition of GPR81 by 3-OBA suppressed HCC cell proliferation and invasion, reduced tumor volume and weight, and downregulated PD-L1 expression. Furthermore, CTL and DC activity in the spleen and tumors increased, while MDSC activity decreased. This study confirms that GPR81 plays an important role in both inflammation and tumorigenesis, suggesting that GPR81 may serve as a bridge in the transformation of inflammation into cancer. Modulating GPR81 activity may provide a novel therapeutic strategy for hepatitis and cancer. Full article

Utilizing deep eutectic solvents (DESs) combined with microwave-assisted extraction (MAE) provides a sustainable method for extracting bioactive compounds from the macroalgae Ascophyllum nodosum and Laminaria hyperborea. Two DES formulations, choline chloride/lactic acid (ChCl/LA) and sodium acetate/lactic acid (AcNa/LA), were evaluated under varying extraction conditions. For L. hyperborea, ChCl/LA at 150 °C for 10 min yielded a total phenolic content (TPC) of 15.34 mg GAE/g DW, with antioxidant activities measured by DPPH (34.55 mg TE/g DW) and ABTS (27.06 mg TE/g DW). Extending the extraction to 20 min at 130 °C increased the TPC to 19.12 mg GAE/g DW. A. nodosum exhibited higher bioactivity, with the TPC reaching 47.51 mg GAE/g DW under the same conditions. High-performance liquid chromatography (HPLC) identified significant phenolics such as 3,4-dihydroxybenzoic acid (678.05 µg/g DW) and vanillin (6718.5 µg/g DW). Antimicrobial assays revealed strong inhibition (zones > 20 mm) against Clostridium perfringens, moderate activity against Staphylococcus aureus, and selective activity against Escherichia coli. FT-IR confirmed the presence of phenolics, polysaccharides, and lipids. Thermal and structural characterization revealed that A. nodosum residue showed an amorphous structure, while L. hyperborea retained crystallinity with decomposition profiles indicating potential bioenergy potential. SEM images revealed significant cell wall disruption correlating with extraction efficiency. These results demonstrate DES–MAE as an effective, green strategy for producing high-value algal extracts and valorizing residual biomass for biotechnological applications. Full article

The rising demand for sustainable and health-promoting foods has encouraged the development of tempeh from non-soy plant materials. This study investigated tempeh alternatives made from sorghum, proso millet, white bean, buckwheat, yellow pea, and quinoa, focusing on their phenolic acid (PA) content. Seventeen PAs and two flavan-3-ols were quantified using LC-MS/MS in free, conjugated, and insoluble forms, and total phenolic content (TPC) was determined using the Folin–Ciocalteu assay. Four PAs—shikimic acid, 3-hydroxycinnamic acid, 3,5-dihydroxybenzoic acid, and 2-hydroxycinnamic acid—were not detected. Solid-state fermentation increased the total PA (TPA) content by an average of 11.3%, reaching 160.6 µg/g, with the most significant rise in conjugated and insoluble fractions. The highest TPA values were observed in sorghum-based tempeh, particularly quinoa:sorghum (2:1; 293 µg/g), sorghum:yellow pea (2:1; 277.6 µg/g), and buckwheat:sorghum (1:1; 271 µg/g). The most abundant PAs were ferulic (18 µg/g), vanillic (14.6 µg/g), 3,4-dihydroxybenzoic (8 µg/g), and caffeic acids (6.7 µg/g). TPC values reached up to 9.51 mg GAE/g in tempeh samples. These findings support the use of non-soy substrates to develop nutritious, allergen-free, gluten-free tempeh products with enhanced phenolic profiles and functional food potential. Full article

Background/Objectives: Metabolic dysfunction-associated steatotic liver disease (MASLD) is the most prevalent chronic liver disease worldwide, contributing to metabolic dysfunction and increased healthcare costs. The green Mediterranean diet reduces intrahepatic fat and elevates the plasma levels of 2,5-dihydroxybenzoic acid (2,5-DHBA), suggesting a mechanistic role for 2,5-DHBA in hepatic lipid metabolism. This study aimed to evaluate the therapeutic potential of 2,5-DHBA in MASLD and elucidate its molecular mechanism. Methods: Lipid accumulation was assessed in oleic acid-treated HepG2 cells and a high-fat diet (HFD)-induced MASLD mouse model. RNA sequencing, molecular docking, and immunohistochemical staining were performed to investigate the molecular mechanisms, focusing on the chemokine (C-C motif) ligand 2 (CCL2)–CCL2 receptor (CCR2) axis. Results: 2,5-DHBA significantly reduced hepatic lipid accumulation in both HepG2 cells and HFD-fed mice in a dose-dependent manner. RNA sequencing revealed the marked downregulation of CCL2, a key proinflammatory mediator in MASLD pathogenesis. Molecular docking predicted that 2,5-DHBA competed with CCL2 for binding at the CCR2 axis. Immunohistochemistry further confirmed that 2,5-DHBA treatment lowered hepatic CCL2 expression, suppressed nuclear factor-κB activation, and reduced inflammatory cell infiltration. These findings suggest that 2,5-DHBA exerted anti-steatotic effects by modulating the CCL2-CCR2 signaling pathway. Conclusions: This is the first study to demonstrate that 2,5-DHBA attenuates hepatic steatosis via targeting the CCL2-CCR2 axis. These findings highlight its potential as a novel nutraceutical strategy for MASLD treatment. Full article

Rainwater needs to be recognized as a natural water source for domestic use, but finding viable processes to remove its contaminants is essential. The aim of this work was to compare the UV/H₂O₂ and UV/Fenton-like processes for the oxidation of 3,5-dihydroxybenzoic acid (3,5-DHBA) in rainwater. The reactions were assessed using ultraviolet-visible (UV-Vis) and molecular fluorescence spectroscopies, and the results showed the formation of new and similar chromophoric compounds in both processes, which were subsequently degraded. At environmentally relevant concentrations of chemical oxidants, namely H₂O₂ at 10⁻⁴ M, the chromophoric organic compounds in solution were degraded within 24 h by the UV/H₂O₂ process and within 4 h by the UV/Fenton-like process. However, when the concentration of H₂O₂ was increased by one order of magnitude for the UV/H₂O₂ process (from 10⁻⁴ M to 10⁻³ M), oxidation rates were similar and nearly complete after 4 h for both UV/H₂O₂ and UV/Fenton-like processes. These findings highlight that the presence of more oxidizing agents in the oxidation system improves the synergistic effect, leading to a greater contribution of the free radical oxidation pathway, particularly through hydroxyl radicals. Thus, by increasing the concentration of H₂O₂ in the UV/H₂O₂ process to 10⁻³ M, it was possible to achieve a similar level of oxidation (close to 100% after 4 h, as indicated by a decrease in fluorescence intensity) as the UV/Fenton-like process at environmentally relevant concentrations (10⁻⁴ M), but using fewer chemical reactants, since UV/H₂O₂ process does not require Fe(III) as catalyst and oxidant. Therefore, the UV/H₂O₂ process can be considered a simpler and cleaner process for removing organic contaminants from rainwater. Full article

In our previous study, a taste sensor modified with 3-bromo-2,6-dihydroxybenzoic acid (3-Br-2,6-DHBA) exhibited significant responses to xanthine-based substances, suggesting an allosteric detection mechanism. This study investigates the potential of the 3-Br-2,6-DHBA-modified sensor membrane for detecting other drug classes. Eleven structurally diverse drugs—including caffeine, antibiotics, antivirals, analgesic-antipyretics from the WHO Model List of Essential Medicines for Children—were tested, as they were previously undetectable by a conventional bitterness sensor. Among them, amoxicillin, an oral broad-spectrum penicillin, and cefalexin, an oral cephalosporin, elicited significantly higher sensor responses when 3-Br-2,6-DHBA-modified membrane was used. To further examine this response, experiments were conducted using membranes modified with 3-Br-2,6-DHBA, 2,6-dihydroxybenzoic acid (2,6-DHBA), and benzoic acid. These tests confirmed that only 3-Br-2,6-DHBA-modified membrane produced significant responses to amoxicillin and cefalexin, suggesting that hydroxyl groups in 3-Br-2,6-DHBA contribute to allosteric effects via hydrogen bonding. Additional tests demonstrated higher responses for cefaclor and cefdinir, both oral cephalosporins. The interaction between 3-Br-2,6-DHBA and the beta-lactam ring, as well as adjacent five- or six-membered rings in amoxicillin and several oral cephalosporins, likely enables allosteric detection by stacking via π electron, hydrophobilc interaction, and hydrogen bonding. In conclusion, the 3-Br-2,6-DHBA-modified sensor membrane effectively detects amoxicillin and oral cephalosporins via allosteric mechanism. Full article

We demonstrate a rapid and sensitive boron detection method through current amplification mediated by supramolecular interaction. Oxidation peak currents obtained by cyclic voltammetry (CV) measurements of a ferrocene/catechol-functionalized β-cyclodextrin inclusion complex were amplified through an EC’ reaction (where EC’ denotes an electrochemical step followed by a catalytic chemical step). However, the amplified current was decreased by boric acid (the primary form of boron in water) addition at pH 8.6 owing to interactions of boron with the cis-diol structure of dihydroxybenzoic acid-β-cyclodextrin and ferrocene for ester formation. We determined the optimum CyD functionalization sites and measurement conditions and obtained a limit of detection of 0.16 mg B L⁻¹ for ferrocene/3,4-dihydroxybenzoic acid-β-cyclodextrin (Fc/3,4-DHBA-β-CyD). The binding constant (assuming a 1:1 binding model) for the interaction between Fc/3,4-DHBA-β-CyD and boric acid was estimated to be approximately 1500 M⁻¹. Boron concentrations in spiked real samples showed good recoveries and linear calibration curves. The electrochemical response of this system was not significantly affected by the presence of other anions or cations. We also found that an aqueous solution of 3,4-DHBA-β-CyD remained stable for at least 112 days. Full article

In this study, the compositional shifts in free and bound phenolic compounds of Morinda citrifolia L. (Noni) processed by different drying methods were investigated. Twenty-seven phenolic compounds, predominantly rutin and quinic acid, were discovered in fresh Noni fruit. Vacuum freeze-drying retained the highest free phenolic content, with rutin (1809.83 mg/kg DW) and quinic acid (198.72 mg/kg DW) as the primary constituents, while bound phenolics were dominated by benzoic acid (35.56 mg/kg DW). Hot-air drying reduced the free phenolics by 51.59% (80% methanol) and the bound phenolics (base hydrolysis) by 35.55%, with a significant degradation of rutin and quinic acid. Microwave drying similarly decreased the free phenolics, though the caffeic acid increased to 46.45 mg/kg DW due to the thermal stability. Bound phenolics showed the highest content (alkaline hydrolysis) in fresh fruits, primarily benzoic acid (220.67 mg/kg DW) and rutin (77.02 mg/kg DW), surpassing the acid/enzyme methods. While vacuum freeze-drying effectively preserved the free phenolics, thermal methods (hot-air/microwave drying) promoted the release of quercetin (free phenols) and 3,4-dihydroxybenzoic acid (bound phenols). The findings of this study elucidate the species-specific compositional dynamics of phenolic compounds under different drying regimes, while providing quantitative guidelines for advancing the understanding of the underlying health-promoting phytochemical profiles of Noni. Full article

Plants of the genus Betula pendula (Betulaceae) have been used as antiseptics in the treatment of skin diseases. While the bark of this plant has so far been used as a medicinal raw material, our research has revealed that the leaves are an extremely valuable source of health-promoting bioactive compounds, especially polyphenolic compounds. The antioxidant activity of a novel lotion from the Betula pendula leaf and its phenolic content was determined using the DPPH and FRAP assays and the Folin–Ciocalteu method, and a qualitative analysis was conducted using the GC-MS method. The antiseptic preparation was also subjected to HPLC analysis for an assessment of the phenolic acids content (gallic, 3,4-dihydroxybenzoic, 2,5-dihydroxybenzoic, chlorogenic, and 3-hydroxybenzoic acids). The shake flask method was used to determine the partition coefficient of the lotion to assess its lipophilicity. A new ethanol-reduced antiseptic lotion containing antimicrobial ingredients was evaluated using standard killing methods (EN 13697:2019 and EN 13697:2015) at 60 ± 10 s and 300 ± 10 s against six microorganisms. The aim of these tests was to determine whether the formulation exhibited performance like a traditional ethanol. In addition, hand disinfection tests were also carried out using the preparation obtained from B. pendula leaves in accordance with EN 1500:2013, using Escherichia coli strain K12 NCTC 10538. According to the results, high polyphenol contents (52 ± 2 mg GAE/g dry raw material) and other antioxidant and biologically active substances identified by GC-MS (phytol, alpha-tocospiro B, sitosterol, or dilauryl thiodipropionate) can be thought of as the parameters responsible for the effective activity of the preparation derived from birch leaves. Gallic acid and 3,4-dihydroxybenzoic acid had the greatest concentrations among the phenolic acids examined. The mean 60 s kill time was >99.9 for the new antiseptic against Staphylococcus aureus ATCC 6538 (at a concentration of 30 g/100 mL) and Pseudomonas aeruginosa ATCC 15442 (at a concentration of 40 g/100 mL), while ethanol showed the same reduction for these microorganisms at a concentration of 80 g/100 mL. Full article

In traditional Chinese medicine, the root bark and leaves of Schima argentea are utilized to treat dysentery, parasitic infections, and digestive disorders. In this study, the n-butanol extract of S. argentea (NBA) exhibited potent antioxidant properties, protecting HaCaT cells from UVB-induced damage, and was abundant in phenolic and flavonoid compounds. Using UPLC-QTOF-MS analysis, several antioxidants within NBA were identified. Among these, 3,4-dihydroxybenzoic acid, (+)-catechin, and procyanidin B2 effectively reduced ROS levels after 1 h post-UVB treatment (225 mJ/cm²). Notably, all three compounds significantly decreased the phosphorylation of p38 and JNK in a dose-dependent manner. Additionally, the cell survival rate of these compounds was assessed after 12 h post-UVB treatment (225 mJ/cm²). Both 3,4-dihydroxybenzoic acid and (+)-catechin significantly prevented UVB-induced apoptosis in HaCaT cells, as evidenced by MTT, Hoechst, Calcein/PI staining, and flow cytometry analyses. Proteomic analysis revealed that 3,4-dihydroxybenzoic acid achieved photoprotection by downregulating c-Fos and Jun and modulating cell cycle proteins, while (+)-catechin promoted cell repair through the PI3K-Akt and Wnt signaling pathways. These results demonstrated that both compounds can directly absorb UVB, scavenge ROS, and provide cell photoprotection by modulating multiple signaling pathways. The n-butanol extract of S. argentea holds promising potential for future medical applications. Full article

Inflammatory bowel disease (IBD) is characterized by uncontrolled, chronic relapsing inflammation in the gastrointestinal tract and has become a global healthcare problem. Here, we aimed to illustrate the anti-inflammatory activity and the underlying mechanism of methyl 3-bromo-4,5-dihydroxybenzoate (MBD), a compound derived from marine organisms, especially in IBD, using a zebrafish model. The results indicated that MBD could inhibit the inflammatory responses induced by CuSO₄, tail amputation and LPS in zebrafish. Furthermore, MBD notably inhibited the intestinal migration of immune cells, enhanced the integrity of the gut mucosal barrier and improved intestinal peristalsis function in a zebrafish IBD model induced by trinitro-benzene-sulfonic acid (TNBS). In addition, MBD could inhibit ROS elevation induced by TNBS. Network pharmacology analysis, molecular docking, transcriptomics sequencing and RT-PCR were conducted to investigate the potential mechanism. The results showed that MBD could regulate the TLR/NF-κB pathways by inhibiting the mRNA expression of TNF-α, NF-κB, IL-1, IL-1β, IL6, AP1, IFNγ, IKKβ, MyD88, STAT3, TRAF1, TRAF6, NLRP3, NOD2, TLR3 and TLR4, and promoting the mRNA expression of IL4, IκBα and Bcl-2. In conclusion, these findings indicate that MBD could be a potential candidate for the treatment of IBD. Full article

Accelerated photooxidation of salicylic acid (SA) was performed using UV radiation and hydrogen peroxide. HPLC-MS analysis showed that the primary intermediates are 2,5-dihydroxybenzoic acid, 2,3-dihydroxybenzoic acid, pyrocatechol, and phenol. Deeper oxidation leads to low molecular weight aliphatic acids, such as maleic, fumaric, and glyoxylic. The photooxidation of the main intermediates was carried out in the same conditions. The degradation of SA and its main intermediates follows first-order reaction kinetics. In the case of UV irradiation alone, photodegradation of 2,5-dihydroxybenzoic acid is slightly faster (reaction rate constant is 0.007 min⁻¹) compared to SA (0.0052 min⁻¹). Other products degrade more slowly than SA. Hydrogen peroxide, in concentrations of 1.8–8.8 mM, accelerates the photodegradation of salicylic acid and intermediate products. An ecotoxicological evaluation of SA and the main products was performed using the EPI Suite^TM software. The overall persistence (P_OV) and long-range transport potential (LRTP) of all transformation products were assessed using OECD P_OV and the LRTP screening tool. Salicylic acid and its transformation products have low toxicity. Due to their high solubility, these contaminants can travel considerable distances in the aquatic environment. SA and phenol have LRTP values of 156–190 km. Other products can travel shorter distances (less than 100 km). Full article