Search Results (46)

Myogenesis is a tightly regulated process essential for embryonic development, postnatal growth, and muscle regeneration. We recently identified that cytokinins (CTKs), a class of adenine-derived signaling molecules originally characterized in plants, are present in cultured skeletal muscle cells. The most abundant type of cytokinins detected within cultured muscle cells was isopentenyladenine (iP) in its nucleotide, riboside, and free base derivatives. The purpose of this study was to determine whether CTKs are also present in regenerating muscle tissue in vivo and to characterize the effects of iP and its riboside form, isopentenyladenosine (iPR), on muscle cell proliferation and differentiation. These effects were observed relative to adenine and adenosine, and to a second class of cytokinins with a large aromatic side chain, kinetin (the free base), and kinetin riboside. Cardiotoxin was used to induce muscle injury and repair processes in the gastrocnemius of 3- and 12-month-old mice. Samples were collected 3- and 7 days post-injury for ultra high-performance liquid chromatography tandem mass spectrometry with electrospray ionization (UHPLC-(ESI+)-HRMS/MS). Four CTKs (N⁶-benzyladenine (BA), dihydrozeatin-9-N-glucoside (DZ9G), isopentenyladenosine (iPR), and 2-methylthio-isopentenyladenosine (2-MeSiPR) were detected. 2-MeSiPR levels were significantly influenced by aging, as this CTK was increased in response to injury only in the younger mice. Treatment of C2C12 myoblasts with 10 µM of isopentenyladenosine (iPR) or kinetin riboside reduced cell proliferation, whereas iP (the free base) increased proliferation in a biphasic response. During differentiation, both iPR and kinetin riboside impaired myotube formation, while the free-base forms of iP and kinetin had no effect. Our data establishes that CTKs are present within muscle tissue and highly responsive to injury and aging. Furthermore, the biological activities of CTKs in muscle cells are influenced by structural modifications, including riboside conjugation and side chain composition. Understanding these differences provides insight into the distinct roles of CTKs in muscle cell metabolism and differentiation, offering potential implications for the use of exogenous CTKs in muscle biology and regenerative medicine. Full article

Background/Objectives: Hypericum cerastoides (Spach) N. Robson is a lesser-known species with potential pharmacological importance. This study aimed to profile phenolic compounds in its aerial parts and assess biological activities of isolated constituents, focusing on radical-scavenging, anti-α-glucosidase, and pro-lipase effects. Methods: Phenolic compounds from H. cerastoides aerial parts were dereplicated via UHPLC-HRMS/MS. The structures of isolated compounds were determined using spectroscopic methods (1D and 2D NMR, UV, and HRMS-ESI). Radical-scavenging was evaluated by DPPH and ABTS assays; anti-α-glucosidase and pro-lipase activities were measured by LC-MS. Results: UHPLC-HRMS profiling of a hydroalcoholic extract tentatively identified and quantified 39 phenolic compounds, mainly flavonoids and hydroxycinnamic acid derivatives. Furthermore, two new phenolic compounds, namely hypercerastoside A (HC4) and hypercerastoside B (HC6), together with three known compounds, coumaroylquinic acid (HC1), myricetin-3-O-glycoside (HC2), and myricetin-3-O-galactoside (HC3), as well as two artifacts, namely methyl ester of chlorogenic acid (HC5) and hypercerastoside C (HC7), were isolated from the ethylacetate extract of the aerial parts of title plant. Compounds HC2, HC3, and HC5 displayed the highest radical-scavenging activity. The anti-α-glucosidase test showed that compounds HC1 (IC₅₀ = 44 µM) and HC3 (IC₅₀ = 206 µM) possessed similar activity to acarbose (IC₅₀ = 206 µM). Myricetin glycosides HC2 and HC3 enhanced lipase activity fivefold at 200 µM. Conclusions: H. cerastoides is a promising source of bioactive phenolic compounds with significant radical-scavenging and enzyme-modulating activities. These preliminary findings support further exploration of its therapeutic potential, especially for oxidative stress-related disorders, type 2 diabetes, and cachexia. Full article

Eperua oleifera Ducke (Fabaceae), commonly known as copaíba-jacaré, is traditionally used for therapeutic purposes, like Copaifera oleoresins. Previous GC-MS studies reported its chemical composition as mainly composed of diterpenic acids, consistent with species of the same genus. Although GC-MS remains widely used for comparing compound retention times and fragmentation patterns, its application to diterpenic acids requires a derivatization step to form methyl esters due to the poor chromatographic performance of carboxylic acids on methyl silicone stationary phases. This step may lead to misinterpretations, especially considering recent findings of naturally occurring methyl esters in oleoresins that may co-elute with derivatized acids. This study aimed to apply more sensitive analytical techniques to identify both target and untargeted compounds. The resin of E. oleifera was analyzed by GC-MS to assess the presence of volatile components. Additionally, UHPLC-HRMS was employed using full-scan MS, data-dependent acquisition (DDA), and parallel reaction monitoring (PRM) in both positive and negative ESI modes. GC-MS confirmed the absence of volatile sesquiterpenes, classifying E. oleifera as a resin. Targeted UHPLC-HRMS detected natural methyl esters of diterpenic acids, while untargeted analysis using Compound Discoverer 3.3 software revealed flavonoids and phenolic compounds not previously reported. These findings support the application of UHPLC-HRMS as a powerful tool in phytochemical studies. Full article

Mauritia flexuosa, commonly known as “canangucha,” holds significant nutritional and economic value in the Amazon region. While its pulp is widely utilized in local food products, the seed or kernel is largely underutilized. This study investigated the proximal and phytochemical composition of M. flexuosa, alongside its biological properties, specifically focusing on the hypoglycemic activity of an ethanolic extract from M. flexuosa seeds (MFSs). Proximal analysis revealed that MFSs are a notable source of crude fiber (28.4%) and a moderate source of protein (9.1%). Phytochemical screening indicated a high total polyphenol content (123.4 mg gallic acid equivalents/100 mg dry weight) and substantial antiradical capacity against the ABTS radical (IC₅₀ = 171.86 µg/mL). Notably, MFS ethanolic extracts exhibited significant in vitro antihyperglycemic activity via inhibiting α-amylase and α-glucosidase enzymes, demonstrating comparable inhibition to acarbose at higher concentrations. This hypoglycemic effect was further corroborated in an in vivo rat model with induced diabetes, where the administration of 100 mg/kg of MFS ethanolic extract significantly reduced blood glucose levels compared to the diabetic control group (p < 0.05). A moderate antihypertensive effect was observed at a concentration of 150 mg/kg, correlating with ACE inhibition. High-performance liquid chromatography–mass spectrometry (UHPLC-ESI-HRMS) analysis of the seed extract identified phenolic compounds including ellagic, p-coumaric, and chlorogenic acids, as well as flavonoids such as quercetin, myricetin, and epicatechin. This study provides the first evidence of the hypoglycemic activity of MFSs, offering valuable insights into their phytochemistry and potential therapeutic applications. Full article

Phenolic acids are contained in grape pomace, mostly in a conjugate form, and can be a natural source of building blocks if they are efficiently hydrolyzed and extracted from the natural matrix. In this study, a comparative study based on the spectrophotometric evaluation of total phenolic content, hydroxycinnamic acid content, and anthocyanin content was performed on different carboxylic-acid-based NADES with different heating sources. Moreover, a quali–quantitative characterization of the bioactive molecules extracted was performed using UHPLC-ESI-HRMS. We found that the nature of the acidic component of the DES was crucial in selecting the family of molecules to be extracted; ChCl/oxalic acid 1:1 NADES, when combined with MAE at 100 °C, is the best medium for the in situ hydrolysis and extraction of phenolic acids from grape pomace. The ORAC test performed on natural extracts with and without NADES revealed a role for NADES components in antioxidant activity against the ROS of extracted bioactive phenols. Full article

This study aim to compare the extraction yield, antioxidant potential, and chemical composition of Chaetoceros costatus extracted with acetone, ethanol, and hexane. The freeze-dried diatom biomass was extracted by ultrasonication for 1 h at 40 °C. The antioxidant capacity was determined using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and oxygen radical absorbance capacity (ORAC), while the chemical profiles of the extracts were analyzed using high-performance liquid chromatography with high-resolution mass spectrometry with electrospray ionization (UHPLC-ESI-HRMS). The ORAC assay showed a 27% higher activity of the acetone extract, while the DPPH assay showed almost 3-fold higher DPPH inhibition. Pigments, fatty acids, sterols, and their derivatives were identified in all extracts. The chemical composition of ethanolic and acetonic extracts did not differ significantly, and hexane yielded the fewest compounds. The results of this study will contribute to extraction challenges that limit biotechnological application and exploitation of diatoms. Full article

Several polyphenol-rich Terminalia species (Combretaceae) are known to accelerate wound healing. Recently, the Omani medicinal plant Anogeissus dhofarica (now Terminalia dhofarica) was attributed to the genus Terminalia based on phylogenetic studies. Leaves, bark, and extracts of T. dhofarica are traditionally used for various medicinal purposes, including wound treatment and personal hygiene. In the present study, the phytochemical profile of leaves from T. dhofarica was evaluated by ultra-high-performance liquid chromatography coupled with electrospray ionization high-resolution mass spectrometry (UHPLC-ESI-HRMS) and nuclear magnetic resonance (NMR) spectroscopy. Simple phenolics, polyphenolics (e.g., flavonoids and tannins) and their glucosides were characterized as major metabolite classes. In addition, 20 phenolics were isolated and structurally identified. Nine of these compounds were never described before for T. dhofarica. For the first time, we provide complete NMR data for 1-O-galloyl-6-O-p-coumaroyl-d-glucose (1). Biological screening demonstrated moderate efficacy against the Gram-negative bacterium Aliivibrio fischeri, the phytopathogenic fungus Septoria tritici, and the oomycete Phytophthora infestans. In summary, the data expand the knowledge of the phytochemistry of the underexplored species T. dhofarica and underscore its potential for therapeutic applications, particularly in the context of traditional medicine. Full article

Background/Objectives: This study evaluates the effect of Mn(II) and Co(II) ions on the production of anti-Candida metabolites by the endophytic fungus Aspergillus sp., isolated from Dizygostemon riparius. The objective was to identify metal-induced secondary metabolites with antifungal potential against drug-resistant Candida species. Methods: Aspergillus sp. was cultivated in Czapek agar supplemented with MnCl₂ (400 µM) or CoCl₂ (200 µM). Metabolite profiles were analyzed using UHPLC-DAD and LC-ESI-HRMS, followed by structural elucidation via NMR. Antifungal and biofilm inhibition activities were tested against Candida albicans and Candida parapsilosis. Toxicity was assessed using Tenebrio molitor larvae. Results: Key metabolites, including pyrophen, penicillquei B, and fonsecinone B, demonstrated antifungal activity with MIC values of 4.37–280.61 µg/mL. Fonsecinone B exhibited superior biofilm inhibition, surpassing fluconazole in reducing biofilm biomass and viability. In vivo assays showed low toxicity, with survival rates above 80% at 2× MIC/kg. Conclusions: Mn(II) and Co(II) significantly modulated the production of antifungal metabolites in Aspergillus sp. Fonsecinone B emerged as a promising candidate for antifungal therapy due to its potent activity and low toxicity. These findings support further investigation into the therapeutic potential of metal-induced fungal metabolites for combating drug-resistant Candida infections. Full article

Magneto-priming (MP) has been reported as a sustainable method to enhance crop yield and resistance to adverse environmental conditions; however, any physiochemical basis for these findings remains unavailable. In the present study, soybean (Glycine max) seeds (n = 232) were exposed to a static magnetic field (SMF) of 150–205 mT for 60 min. The effect of SMF exposure on seedling mass, hypocotyl length, radicleemergence rate, total seedling height, leaf area, chlorophyl content, and physiological attributes was evaluated. Differential analysis of the cytokinin (CK) profile of seedling tissues was achieved using ultra-high-performance liquid chromatography coupled with electrospray-ionization high-resolution tandem mass spectrometry (UHPLC-(ESI)-HRMS/MS). Results indicate that MP seedlings achieved radicle emergence earlier; however, no other statistically significant differences could be established. The absence in significant improvement in the agronomic and physiological attributes measured is in stark contrast to the results reported in other studies. Nonetheless, significant differences were found in the concentrations of trans-Zeatin (tZ) and cis-Zeatin (cZ) between control and MP plants. These results are incongruent as it would be assumed that insignificant differences in agronomic and physiological properties would align with phytohormone profiles. Future work should be performed to determine if there are any meaningful bioeffects that can be assigned to MP-generated hormone changes. Full article

A new method for simultaneous determination by liquid chromatography coupled with high resolution mass spectrometry (UHPLC-HRMS/MS) of 14 paralytic shellfish poisoning toxins (PSP), that is, Saxitoxin, Neosaxitoxin, Gonyautoxins and their respective variants, in bivalve molluscs, is herein described. The samples were extracted by acetic acid solution, then analysed by UHPLC coupled with a Q-Exactive Orbitrap Plus high resolution mass spectrometer, by electrospray ionization mode (ESI) with no further clean up step. The analysis was carried out by monitoring both the exact mass of the molecular precursor ion of each compound (in mass scan mode, resolution at 70,000 FWHM) and its respective fragmentation patterns (two product ions) with mass accuracy greater than 5 ppm. The analytical performance of the method was evaluated calculating trueness, as mean recoveries of each biotoxin, between 77.8% and 111.9%, a within-laboratory reproducibility (RSD_R) between 3.6% and 12.2%, the specificity, the linearity of detector response, and the ruggedness for slight changes The results of the validation study demonstrate this method fits for the purposes of the official control of PSP toxins in molluscs. The results of two years of monitoring in local mussel farms are also reported, showing that no significant concerns for food safety in the monitored productions. Full article

Trichoderma spp. are filamentous fungi generally observed in nature, which are widely marketed as biocontrol agents. The secondary metabolites produced have obtained special attention since they possess attractive chemical structures with a broad spectrum of biological activities. In Cuba, the species of Trichoderma have been commercially applied for the control of several phytopathogens to protect agricultural crops, but few studies have been carried out to detect and characterize the production of metabolites with biological activity. The strain Trichoderma harzianum LBAT-53 was subjected to an antifungal in vitro assay against Fusarium oxysporum f.sp. cubense by dual culture and volatile metabolite assays and fermented in PDB under constant agitation conditions. The ethyl acetate crude extract was obtained by liquid–liquid extraction. The fungal extract was investigated for the composition of secondary metabolites through chemical screening and ultrahigh performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) in negative ionization mode. As a result, LBAT-53 showed antagonistic activity in vitro (Class 2) against the pathogen evaluated in direct confrontation (76.9% of inhibition in 10 days) and by volatile metabolites (<40% in 7 days). Furthermore, seven low-molecular-weight phenolic compounds, including chrysophanol, phomarin, endocrocin, and trichophenol A, among others, were identified using UHPLC-ESI-MS/MS. This study is the first work on the characterization of secondary metabolites produced by the commercially applied strain LBAT-53, which is a promising source of bioactive compounds. These results provide a better understanding of the metabolism of this fungus, which is widely used in Cuba as biopesticides in agriculture pest control. Full article

Phyllanthus emblica L. fruits (PEFs) were processed by ultra-pressure (UHP) treatment and then extracted by the ultrasonic-assisted extraction method. The influence of UHP on the phenolic composition, enzyme inhibitory activity and antioxidant activity of the free, esterified, and bound phenolic fractions from PEFs were compared. UHP pretreatment of PEFs significantly increased the total phenolic and flavonoid contents (p < 0.05). A total of 24 chemical compositions were characterized in normal and UHP-treated PEFs by UHPLC-ESI-HRMS/MS. Compared with normal PEFs, these three different phenolic fractions had stronger antioxidant activities and inhibitory effects on the intracellular reactive oxygen species (ROS) production in H₂O₂-induced HepG2 cells (p < 0.05). The ROS inhibition might be due to an up-regulation of the expressions of superoxide dismutase (SOD) and glutathione (GSH) activities. In addition, these three different phenolic fractions also significantly inhibited the activities of metabolic enzymes, including α-glucosidase, α-amylase and pancreatic lipase. This work may provide some insights into the potential economics and applications of PEFs in food and nutraceutical industries. Full article

Distillers’ grains are rich in protein and constitute a high-quality source of various bioactive peptides. The purpose of this study is to identify novel bioactive peptides with α-glucosidase inhibitory, antioxidant, and insulin resistance-ameliorating effects from distiller’s grains protein hydrolysate. Three novel peptides (YPLPR, AFEPLR, and NDPF) showed good potential bioactivities, and the YPLPR peptide had the strongest bioactivities, whose IC₅₀ values towards α-glucosidase inhibition, radical scavenging rates of 2,2′-azino-bis (3-ethylbenzothiazoline-6- sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) were about 5.31 mmol/L, 6.05 mmol/L, and 7.94 mmol/L, respectively. The glucose consumption of HepG2 cells treated with YPLPR increased significantly under insulin resistance condition. Moreover, the YPLPR peptide also had a good scavenging effect on intracellular reactive oxygen species (ROS) induced by H₂O₂ (the relative contents: 102.35% vs. 100%). Molecular docking results showed that these peptides could stably combine with α-glucosidase, ABTS, and DPPH free radicals, as well as related targets of the insulin signaling pathway through hydrogen bonding and van der Waals forces. This research presents a potentially valuable natural resource for reducing oxidative stress damage and regulating blood glucose in diabetes, thereby increasing the usage of distillers’ grains peptides and boosting their economic worth. Full article

The limited availability of phosphate, nitrogen and silicon in the growth media affects the growth, cellular processes, and metabolism of diatoms. Silicon deficiency primarily affects diatom morphology, while phosphate deficiency reduces the production of nucleic acids and phospholipids. Differences in pigment and protein composition are mainly due to nitrogen deficiency. In this study, Chaetoceros socialis and Chaetoceros costatus were cultured under phosphate, nitrogen, and silicon deprivation conditions. The diatom biomass was collected during the stationary growth phase and extracted with 70% ethanol under ultrasonication. The chemical profiles of the extracts were analyzed by high-performance liquid chromatography with high-resolution mass spectrometry with electrospray ionisation (UHPLC-ESI-HRMS), while the antioxidant capacity was determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and oxygen radical absorbance capacity (ORAC) assays. Pigments, fatty acids, sterols, and derivatives were detected in both species. The total phenolic content in the extracts ranged from 46.25 ± 1.08 to 89.38 ± 6.21 mg of gallic acid equivalent (GAE)/L and from 29.58 ± 1.08 to 54.17 ± 1.18 mg GAE/L. for C. costatus and C. socialis, respectively. Antioxidant activity was higher in C. costatus extracts, especially those obtained from nitrogen-deprived media. The results of this study contribute to the existing knowledge and the ongoing efforts to overcome application and commercialization barriers of microalgae for wide-ranging potential in different industries. Full article

Background and objectives: The incidence of chronic inflammatory pathologies has incrementally increased in recent years, as in the case of inflammatory bowel disease (IBD), which is characterized by intestinal epithelial barrier disruption, increased inflammatory mediator production and excessive tissue injury. Changes in eating habits might have played a key role in this scenario. Therefore, the interest in specific diet development and in functional food formulation has been growing. Phytoextracts from several origins, from plants to waste, enriched in bioactive molecules, alone or combined, might be a resource for the obtainment of an efficient synergistic beneficial. Thus, the aim of this study consists of evaluating the protective effects of artichoke, fenugreek and caigua (AFC) phytoextract original blend. Methods: In order to mimic the intestinal barrier’s inflammatory environment, Caco-2 cells were cultured and polarized on a transwell system and then exposed to a pro-inflammatory cytokine cocktail (TNFα and IL-1β). Before being exposed to an inflammatory stimulus, cells were pre-treated with an AFC digested blend, according to the INFOGEST in vitro static digestion protocol. After digestion, the content of active substances within the blended extract (ACFB) was revealed by UHPLC–ESI–HRMS analysis. The AFC digested extract’s protective effect was evaluated by measuring the transepithelial resistance (TEER) as a marker of barrier integrity and analysing the nuclear factor kappa B (NF-κB) pathway. Results: The TEER values improved in cells which were pre-treated with the AFC blend, relative to inflamed cells, suggesting a regulation in tight junction protein expression and/or localization. The transcription factor p65NF-κB is activated by phosphorylation under cytokine exposure, with a 160% increase in its target COX-2. Moreover, a 40-fold increase in IL-8 release was observed. Interestingly, in cells pre-treated with the AFC blend, the activated p65NF-κB was halved, compared to inflamed cells only. Furthermore, a consequent reduction by about 50% for COX-2 and by 30% for IL-8 was observed. Discussion: Taken together, these results highlight the anti-inflammatory potential of the AFC blend, probably due to the presence of flavonoids such as luteolin, apigenin and chrysin. This experimental evidence suggests that an AFC blend could be a good ingredient for food functionalization if further used in nutritional strategies. Full article