Search Results (43)

Background: This study aimed to analyze metabolic changes in blood samples from patients with confirmed COVID-19 to explore the correlation between metabolomics and cytokines in survivors and non-survivors of SARS-CoV-2 infection. Understanding the complex biochemical and immunometabolic mechanisms underlying SARS-CoV-2 infection is essential for elucidating the pathophysiology and virulence of COVID-19. Methods: This study included 40 hospitalized COVID-19 patients and 40 healthy controls. Serum metabolic profiles were analyzed using ultra-high-pressure liquid chromatography-mass spectrometry (UHPLC-MS), and cytokine levels were measured using ELISA. Results: Our study defined three clear metabolic phenotypes among survivors and non-survivors of COVID-19 compared with healthy controls, which might be related to mortality, severity, and disease burden. A strong relationship was observed between certain inflammatory markers, including IL-1β, IL-2, IFN-β, IFN-γ, IL-17, and GM-CSF, as well as several metabolites, particularly in COVID-19 non-survivors, such as LysoPCs, 3-hydroxykynurenine, and serotonin. Different metabolite-cytokine correlation patterns were observed according to patient outcomes, indicating unique correlations between metabolic and immune responses in survivors and non-survivors. Metabolic phenotypes were associated with clinical outcomes, comorbidities, and sex-related differences. Kynurenine and related metabolites of tryptophan metabolism were closely correlated with COVID-19 severity, age, and mortality. Compared with survivors and healthy controls, non-survivors displayed higher IL-6, together with distinct metabolic changes. These included increased kynurenine through the IDO1 pathway, elevated glucose and lactate reflecting hyperglycolysis and energy stress, and higher xanthosine from purine turnover. Stronger cytokine–metabolite correlations in this group point to tightly linked immunometabolic activation. Conclusions: Metabolomic profiling revealed distinct metabolic phenotypes that could be associated with the severity and inflammation levels of COVID-19. Correlation analysis between metabolites and cytokines demonstrated strong intercorrelations between specific metabolites and cytokines, indicating a strong interrelationship between inflammatory markers and metabolic alterations. Specific metabolic pathways associated with cytokines and their clinical relevance may serve as potential therapeutic targets. Full article

Chestnut (Castanea sativa Mill.) is an edible nut recognized for its nutritional attributes, particularly its elevated levels of carbohydrates (starch) and proteins. Chestnuts are popular for their health-promoting properties and hold significant environmental and economic importance in Europe. During this study, after the characterization of the fruit, attention was directed toward the valorization of chestnut shells, a predominant by-product of industrial chestnut processing that is typically discarded. Valuable bioactive compounds were extracted from the shells using Pressurized Liquid Extraction (PLE), a green, efficient, scalable method. Response surface methodology (RSM) was utilized to determine optimal extraction conditions, identified as 40% v/v ethanol as the solvent at a temperature of 160 °C for 25 min under a constant pressure of 1700 psi. High total polyphenol content (113.68 ± 7.84 mg GAE/g dry weight) and notable antioxidant activity—determined by FRAP (1320.28 ± 34.33 μmol AAE/g dw) and DPPH (708.65 ± 24.8 μmol AAE/g dw) assays—were recorded in the optimized extracts. Ultrahigh-performance liquid chromatography coupled with a hybrid trap ion mobility-quadrupole time-of-flight mass spectrometer (UHPLC-TIMS-QTOF-MS) was applied to further characterize the compound profile, enabling the identification of phenolic and antioxidant compounds. These findings highlight the possibility of using chestnut shell residues as a long-term resource to make valuable products for the food, medicine, cosmetics, and animal feed industries. This study contributes to the advancement of waste valorization strategies and circular bioeconomy approaches. Full article

Pharmaceutical pollution of surface waters has emerged as a significant environmental health concern worldwide. In this study, we investigated the presence and concentration of pharmaceuticals in aquatic environments within Vhembe District Municipality, South Africa. To achieve this, grab samples of water were collected from various locations across the Thulamela Local Municipality encompassing rivers, streams, and dams. A targeted solid-phase extraction method with ultra-high-pressure liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS/MS) was used to screen, detect, and quantify 98 pharmaceutical compounds and caffeine in the collected water samples. The findings revealed the presence of a range of pharmaceutical compounds, including the antiretrovirals nevirapine and lopinavir, the anticonvulsant/antiepileptic carbamazepine, and the analgesic and antipyretic acetaminophen. The central nervous system stimulant caffeine was also detected in various water bodies across the region. The presence and concentrations of the pharmaceuticals varied across different water bodies, with nevirapine present at nine sites out of twenty-one (up to 166 ng/L), lopinavir at two sites (up to 42 ng/L), carbamazepine at one site (21 ng/L), and acetaminophen at two sites with the highest concentration of 427 ng/L. Caffeine was present at 15 sites (up to 975 ng/L). This study provides valuable insights into pharmaceutical pollution in surface water resources from one of South Africa’s rural areas, Vhembe District Municipality. It contributes to the monitoring data required to find sustainable solutions to the problem of pollutants of emerging concern in aquatic environments worldwide. Full article

As a functional extraction medium, natural deep eutectic solvents (NADESs) can dissolve various plant metabolites. Those solvents provide untapped potential for creating novel green extracts with distinctive phytochemical signatures and unique biological activities. This is particularly relevant given the rising need for eco-friendly and sustainable skin care products. The main aim of this work was to optimize the most efficient natural deep eutectic solvents (NADESs) for extracting bioactives from 18 medicinal herbs applicable to the cosmetic industry. Selection of the most potent herbs involved assessing their conventional extracts for tyrosinase inhibition, antioxidant activity, and keratinocyte cytotoxicity. Moreover, we analyzed the phenolic profile using ultra-high-pressure liquid chromatography/mass spectrometry (UHPLC/MS) and spectrophotometric assays such as total phenolic (TPC) and flavonoid content (TFC). Using the COSMO-RS method, we modeled the solubility of 12 phenolics in 64 virtual NADESs and selected the 7 most promising ones for further experimental validation. NADESs, including betaine-urea, betaine-proline, and betaine-lysine, were computationally chosen and demonstrated the highest levels of TPC and antioxidative capacity, as confirmed by in vitro assays. The proposed combination of NADES herbal extracts represents a promising natural constituent for the cosmetic industry. Full article

Microcystins (MCs) and nodularins (NODs) are cyanotoxins that can be found in water bodies during cyanobacterial harmful algal blooms (cyanoHABs). Consumption of water contaminated with cyanotoxins leads to health risks for humans and animals. Herein, corncob-based biochar and activated carbon (AC) were initially investigated for the sorption of six common MC congeners (MC-RR, MC-YR, MC-LR, MC-LA, MC-LW, and MC-LF) and nodularin-R (NOD-R) from spiked water. Biochar was prepared by refluxing commercial corncob with HCl and heating it to 250, 300, or 350 °C. AC was prepared by chemical activation of corncob with H₃PO₄ at 500 °C under a nitrogen atmosphere. Low-temperature nitrogen adsorption measurements confirmed that H₃PO₄-AC has a higher specific surface area (≈1100 m²/g) and total pore volume (≈0.75 cm³/g) than biochar and commercial AC. H₃PO₄-AC showed the maximum efficacy, among all corncob-based sorbents, to remove MCs and NOD-R from water as confirmed by experiments that involved sample analyses by ultrahigh-pressure liquid chromatography-mass spectrometry (UHPLC-MS). The effect of natural organic matter (NOM) on the adsorption of MCs was checked by incubating sorbents with Lake Erie water collected during cyanoHABs from 2020 to 2022. The total concentration (extracellular and intracellular) of studied MC congeners ranged from 1.37 µg/L to 438.51 µg/L and 50 mg of H₃PO₄-AC completely removed them from 3 mL of lake water. The effect of water pH on cyanotoxin adsorption was studied at pH values of 5.5, 7.0, and 8.5 at both a lower (10 μg/L each) and a higher (50 μg/L each) toxin concentration. Removal was influenced by solution pH at both concentrations when using biochar, while only at higher toxin concentration when using H₃PO₄-AC. At higher MC and NOD-R concentrations, competitive adsorption was prominent, and overall, the adsorption increased at acidic pH (5.5). The study results suggest that processed corncobs can remove a significant amount of MCs and NOD-R from water, and the measured sorption capacity of H₃PO₄-AC was ~20 mg of MC-LR and NOD-R per g of this sorbent. Full article

In this study, a Gas chromatography–mass spectrometry (GC–MS) investigation of embryogenic callus and somatic embryo regenerated shoots of Carthamus tinctorius revealed the presence of a variety of sugars, sugar acids, sugar alcohols, fatty acids, organic acids, and amino acids of broad therapeutic value. The in vitro developed inflorescence contained a wide range of active compounds. In embryogenic calluses, important flavonoids like naringenin, myricetin, kaempferol, epicatechin gallate, rutin, pelargonidin, peonidin, and delphinidin were identified. To augment the synthesis of active compounds, the effect of cadmium chloride (CdCl₂) elicitation was tested for various treatments (T1–T4) along with a control (T0). Varying concentrations of CdCl₂ [0.05 mM (T1), 0.10 mM (T2), 0.15 mM (T3), and 0.20 mM (T4)] were added to the MS medium, and flavonoid accumulation was quantified through ultra-high-pressure liquid chromatography–tandem mass spectroscopy (UHPLC–MS/MS). The flavonoids naringenin, kaempferol, epicatechin gallate, pelargonidin, cyanidin, and delphinidin increased by 6.7-, 1.9-, 3.3-, 2.1-, 1.9-, and 4.4-fold, respectively, at T3, whereas quercetin, myricetin, rutin, and peonidin showed a linear increase with the increase in CdCl₂ levels. The impacts of stress markers, i.e., ascorbate peroxidase (APX), catalase (CAT), and superoxide dismutase (SOD), on defense responses in triggering synthesis were also evaluated. The maximum APX and SOD activity was observed at T3, while CAT activity was at its maximum at T2. The impact of elicitor on biochemical attributes like protein, proline, sugar, and malondialdehyde (MDA) content was investigated. The maximum protein, proline, and sugar accumulation was noted at high elicitor dose T4, while the maximum MDA content was noted at T3. These elevated levels of biochemical parameters indicated stress in culture, and the amendment of CdCl₂ in media thus could be a realistic approach for enhancing secondary metabolite synthesis in safflower. Full article

Oligomers are a particular category of non-intentionally added substances (NIAS) that may be present in food contact materials (FCMs), such as polyethylene terephthalate (PET), and consequently migrate into foods. Here, an ultra-high-pressure liquid chromatography quadruple time-of-flight mass spectrometry (UHPLC-qTOF-MS) method was developed for the analysis of 1st series cyclic PET oligomers in virgin olive oil (VOO) following a QuEChERS clean-up protocol. Oligomer migration was evaluated with two different migration experiments using bottles from virgin and recycled PET: one with VOO samples stored in household conditions for a year and one using the food simulant D2 (95% v/v ethanol in water) at 60 °C for 10 days. Calibration curves were constructed with fortified VOO samples, with the LOQs ranging from 10 to 50 µg L⁻¹ and the recoveries ranging from 86.6 to 113.0%. Results showed no migration of PET oligomers in VOO. However, in the simulated study, significant amounts of all oligomers were detected, with the migration of cyclic PET trimers from recycled bottles being the most abundant. Additional substances were tentatively identified as linear derivatives of PET oligomers. Again, open trimer structures in recycled bottles gave the most significant signals. Full article

A duplicate diet collection method was used to estimate dietary exposure to glutamates in children aged 2–5 years in selected provinces of China. Daily duplicate diet samples were collected from 86 healthy toddlers over three consecutive days. Glutamates were analyzed using ultra-high-pressure liquid chromatography–MS/MS (UHPLC–MS/MS). Results showed that the highest glutamates content was found in mixed meals, at 5.12 mg/kg, followed by powdered formula (3.89 mg/kg), and milk and dairy products (2.29 mg/kg). The total mean daily dietary exposure for subjects was 0.20 mg/kg BW, and P95 daily dietary exposure was 0.44 mg/kg BW, both below the acceptable daily intake (ADI) (120 mg/kg BW) recommended by the Joint (FAO/WHO) Expert Committee on Food Additives (JECFA) and the ADI (30 mg/kg BW) set by the European Food Safety Authority (EFSA). Hence it can be considered that glutamates exposure would cause low risk in this group. Full article

This study’s objective was to separate cellulose, hemicellulose, and lignin after high-pressure supercritical carbon dioxide pretreatment for further valorization. The study investigated the supercritical carbon dioxide pretreatment of apple orchard waste at temperatures of 160–200 °C, for 15–45 min, at a pressure of 10 MPa. Response Surface Methodology (RSM) was used to optimize the supercritical process and to improve its efficiency. The change of functional groups during different pretreatment conditions was examined by Fourier transform infrared (FTIR) spectroscopy. Scanning electron microscopy (SEM) and X-ray diffraction (XRD) confirmed the structural changes in the biomass structure before and after pretreatment. A new ultra-high performance liquid chromatography (UHPLC) coupled with an evaporative light scattering detector (ELSD) method was developed and validated for the determination of carbohydrates in the liquid fraction that resulted after pretreatment. The estimated uncertainty of the method ranged from 16.9 to 20.8%. The pre-treatment of high-pressure supercritical CO₂ appears to be an effective and promising technique for the recovery of sugars and secondary by-products without the use of toxic solvents. Full article

Endogenous cardiotonic steroids (CTSs), such as telocinobufagin (TCB) and marinobufagin (MBG) contain a lactone moiety critical to their binding and signaling through the Na⁺/K⁺-ATPase. Their concentrations elevate in response to sodium intake and under volume-expanded conditions. Paraoxonase 3 (PON3) is an enzyme that can hydrolyze lactone substrates. Here, we examine the role of PON3 in regulating CTS levels in a rat model of chronic kidney diseases (CKD). TCB and MBG were extracted from rat urine samples, and the analyses were carried out using ultra-high pressure liquid chromatography–Orbitrap-mass spectrometry (UHPLC-Orbitrap-MS). Ten-week-old Dahl salt-sensitive wild type (SS-WT) and Dahl salt-sensitive PON3 knockout (SS-PON3 KO) rats were maintained on a high-salt diet (8% NaCl) for 8 weeks to initiate salt-sensitive hypertensive renal disease characteristic of this model. CTS extraction recovery from urine >80% was achieved. For animals maintained on a normal chow diet, the baseline amount of TCB excreted in 24 h urine of SS-PON3 KO rats (6.08 ± 1.47 ng/24 h; or 15.09 ± 3.25 pmol) was significantly higher than for SS-WT rats (1.48 ± 0.69 ng/24 h; or 3.67 ± 1.54 pmol, p < 0.05). Similarly, for the same animals, the amount of excreted MBG was higher in the urine of SS-PON3 KO rats (4.74 ± 1.30 ng/24 h versus 1.03 ± 0.25 ng/24 h in SS-WT; or 11.83 ± 2.91 pmol versus 2.57 ± 0.56 pmol in SS-WT, p < 0.05). For animals on a high-salt diet, the SS-PON3 KO rats had significantly increased levels of TCB (714.52 ± 79.46 ng/24 h; or 1774.85 ± 175.55 pmol) compared to SS-WT control (343.84 ± 157.54 ng/24 h; or 854.09 ± 350.02 pmol, p < 0.05), and comparatively higher levels of MBG were measured for SS-PON3 KO (225.55 ± 82.61 ng/24 h; or 563.19 ± 184.5 pmol) versus SS-WT (157.56 ± 85.53 ng/24 h; or 393.43 ± 191.01 pmol, p > 0.05) rats. These findings suggest that the presence and absence of PON3 dramatically affect the level of endogenous CTSs, indicating its potential role in CTS regulation. Full article

Human exposure to BPs is inevitable mostly due to contaminated food. In this preliminary study, for the first time, the presence of bisphenols (BPs) in aqueous humor (AH) collected from 44 patients undergoing cataract surgery was investigated. The measurements were performed using a sensitive ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC–MS/MS). Chromatographic separation was achieved using a reverse-phase column and a gradient elution mode. Multiple reaction monitoring (MRM) was used. The method was validated for bisphenol A (BPA) and bisphenol F (BPF). The limits of quantification (LOQs) of both investigated analytes were 0.25 ng mL⁻¹. The method was linear in the range of 0.25–20.0 ng mL⁻¹ with correlation coefficients (R²) higher than 0.98. Recovery of analytes was in the range of 99.9 to 104.3% and intra-assay and inter-assay precision expressed by relative standard deviations (RSD%) were less than 5%. BPA was detected in 12 AH samples with mean concentrations of 1.41 ng mL⁻¹. BPF was not detected at all. Furthermore, two structural isomers termed BPA-1, and BPA-2 were identified, for the first time, in 40.9% of the AH samples, with almost twice higher mean concentrations of 2.15 ng mL⁻¹, and 2.25 ng mL⁻¹, respectively. The total content of BPs were higher in patients with coexisting ocular pathologies such as glaucoma, age-related macular degeneration (AMD), and diabetes in comparison to cataracts alone. However, the difference between these groups did not reach statistical significance (p > 0.05). Performed investigations indicate the need for further research on a larger population with the aim of knowing the consequences of BPs’ accumulation in AH for visual function. Full article

The third-generation anaplastic lymphoma tyrosine kinase inhibitor (ALK-TKI) lorlatinib has a unique side effect profile that includes hypercholesteremia and hypertriglyceridemia in >80% of lung cancer patients. Here, we tested the hypothesis that lorlatinib might directly promote the accumulation of cholesterol and/or triglycerides in human hepatic cells. We investigated the capacity of the hepatoprotectant silibinin to modify the lipid-modifying activity of lorlatinib. To predict clinically relevant drug–drug interactions if silibinin were used to clinically manage lorlatinib-induced hyperlipidemic effects in hepatic cells, we also explored the capacity of silibinin to interact with and block CYP3A4 activity using in silico computational descriptions and in vitro biochemical assays. A semi-targeted ultrahigh pressure liquid chromatography accurate mass quadrupole time-of-flight mass spectrometry with electrospray ionization (UHPLC-ESI-QTOF-MS/MS)-based lipidomic approach revealed that short-term treatment of hepatic cells with lorlatinib promotes the accumulation of numerous molecular species of cholesteryl esters and triglycerides. Silibinin treatment significantly protected the steady-state lipidome of hepatocytes against the hyperlipidemic actions of lorlatinib. Lipid staining confirmed the ability of lorlatinib to promote neutral lipid overload in hepatocytes upon long-term exposure, which was prevented by co-treatment with silibinin. Computational analyses and cell-free biochemical assays predicted a weak to moderate inhibitory activity of clinically relevant concentrations of silibinin against CYP3A4 when compared with recommended (rosuvastatin) and non-recommended (simvastatin) statins for lorlatinib-associated dyslipidemia. The elevated plasma cholesterol and triglyceride levels in lorlatinib-treated lung cancer patients might involve primary alterations in the hepatic accumulation of lipid intermediates. Silibinin could be clinically explored to reduce the undesirable hyperlipidemic activity of lorlatinib in lung cancer patients. Full article

The content and composition of dietary supplements is of great interest due to their increasing consumption and variety of available brand offered in the market. Accurate determination of vitamins is important for the improvement of dietary supplement quality and nutrition assessments. In this regard, the simultaneous determination of vitamin D3 (calcitriol—CT and cholecalciferol—CHL) and K2 (menaquinone-4—MK-4 and menaquinone-7—MK-7) in dietary supplements was developed by using ultra-high-pressure liquid chromatography (UHPLC). The overall runtime per sample was above 35 min, with the retention times of 2.40, 6.59, 7.06, and 32.6 min for vitamin D3 (CT and CHL) and vitamin K2 (MK-4 and MK-7), respectively. The limits of detection and limits of quantification for the target nutritional compounds ranged between 0.04–0.05 µg/mL, respectively. The validation results indicated that the method had reasonable linearity (R² ≥ 0.9990), good recovery (>82%), satisfactory intra-day precision (≤1.9%) and inter-day precision (≤3.5%), and high selectivity and specificity. The validated UHPLC method was demonstrated to be precise, accurate, and robust for the simultaneous determination of vitamins D3 (CT and CHL) and K2 (MK-4 and MK-7) in dietary supplements. Full article

In this study, functionalized biscuits were prepared through the enrichment of dough with lemon peel and natural antioxidants extracted from lemon pomace. Lemon pomace extract (LP_E) was analyzed for total phenolic content before addition, and then a known concentration of 50 mg kg⁻¹ was used for the formulation of enriched biscuits. Three different biscuit samples were compared to a control biscuit, without the addition of functional ingredients. The main physicochemical, microbiological, and sensory aspects of doughs and biscuits enriched with LP_E were investigated. The enriched biscuits showed higher phenolic content and antioxidant activity than the control one and a longer induction period (IP), which means that the enriched products had a higher intrinsic resistance to lipid oxidation, thanks to the antioxidant effect exerted by the added fresh lemon peel and the LP_E. Furthermore, from a sensorial point of view, they showed suitable acceptability, in terms of appearance, flavor, and aromatic attributes. Thus, results indicated that the incorporation of lemon processing by-products allowed the production of functional enriched biscuits with improved antioxidant properties. Full article

Currently, there is increased interest in finding appropriate food-grade green extraction systems capable of extracting these bioactive compounds from dietary mushrooms for applications in various food, pharmacological, or nutraceutical formulations. Herein, we evaluated a modified Swiss water process (SWP) method using alkaline and acidic pH at low and high temperature under pressurized conditions as a suitable green food grade solvent to obtained extracts enriched with myco-nutrients (dietary phenolics, total antioxidants (TAA), vitamins, and minerals) from Chaga. Ultra-high performance liquid chromatography coupled to high resolution accurate mass tandem mass spectrometry (UHPLC-HRAMS-MS/MS) was used to assess the phenolic compounds and vitamin levels in the extracts, while inductively coupled plasma mass spectrometry (ICP-MS) was used to determine the mineral contents. Over 20 phenolic compounds were quantitatively evaluated in the extracts and the highest total phenolic content (TPC) and total antioxidant activity (TAA) was observed at pH 11.5 at 100 °C. The most abundant phenolic compounds present in Chaga extracts included phenolic acids such as protocatechuic acid 4-glucoside (0.7–1.08 µg/mL), syringic acid (0.62–1.18 µg/mL), and myricetin (0.68–1.3 µg/mL). Vitamins are being reported for the first time in Chaga. Not only, a strong correlation was found for TPC with TAA (r-0.8, <0.0001), but also, with individual phenolics (i.e., Salicylic acid), lipophilic antioxidant activity (LAA), and total antioxidant minerals (TAM). pH 2.5 at 100 °C treatment shows superior effects in extracting the B vitamins whereas pH 2.5 at 60 and 100 °C treatments were outstanding for extraction of total fat-soluble vitamins. Vitamin E content was the highest for the fat-soluble vitamins in the Chaga extract under acidic pH (2.5) and high temp. (100 °C) and ranges between 50 to 175 µg/100 g Chaga. Antioxidant minerals ranged from 85.94 µg/g (pH7 at 100 °C) to 113.86 µg/g DW (pH2.5 at 100 °C). High temperature 100 °C and a pH of 2.5 or 9.5. The treatment of pH 11.5 at 100 °C was the most useful for recovering phenolics and antioxidants from Chaga including several phenolic compounds reported for the first time in Chaga. SWP is being proposed herein for the first time as a novel, green food-grade solvent system for the extraction of myco-nutrients from Chaga and have potential applications as a suitable approach to extract nutrients from other matrices. Chaga extracts enriched with bioactive myconutrients and antioxidants may be suitable for further use or applications in the food and nutraceutical industries. Full article