Search Results (742)

Prenatal alcohol exposure (PAE) is a major public health concern, yet maternal self-reporting remains unreliable. Meconium accumulates alcohol metabolites during the second half of gestation, and ethyl glucuronide (EtG) is considered a stable and specific biomarker of late-pregnancy alcohol exposure. Diapers containing meconium may serve as a noninvasive method for collecting anonymous biological samples, which could help reduce participation bias in studies of this sensitive subject. This pilot study assessed the feasibility of anonymously collecting meconium-containing diapers for subsequent quantitative EtG analysis after extended periods of frozen storage and international transport, and examined intra-pair consistency in samples from the same newborn. Mothers collected their newborns’ diapers, which were then divided into two aliquots (A and B) by a study assistant and stored at −80 °C until analysis. Out of 178 samples collected from 105 infants, 137 samples were analyzable. Eleven samples exceeded the limit of quantification (10 ng/g), while two samples (one pair) surpassed the 30 ng/g cutoff, indicating significant PAE. EtG concentrations showed high intra-pair agreement, supporting the robustness of the biomarker and analytical method. Anonymous diaper-based meconium collection is feasible but operationally demanding. Although the low participation rate and methodological factors, including sample loss, precluded true prevalence estimation, making it exploratory, the detection of quantifiable EtG in 8% of analyzable samples suggests that PAE remains an issue. Full article

Aging is associated with chronic, low-grade inflammation (“inflammaging”), which contributes to neuropsychiatric and neurodegenerative disorders such as depression, Alzheimer’s disease, and Parkinson’s disease. Conventional pharmacotherapies often provide limited benefit in older adults and are further complicated by polypharmacy and drug–drug interactions. Psilocybin, a serotonergic psychedelic acting primarily as a partial agonist at the 5-HT_2A receptor and currently undergoing accelerated clinical development, has emerged as a potential multimodal therapeutic agent addressing these challenges. Acting via its active metabolite psilocin, 5-HT_2A receptor-mediated signaling modulates cortical glutamatergic transmission, enhances tropomyosin receptor kinase B/brain-derived neurotrophic factor (TrkB/BDNF) pathways, and modulates neuroimmune cascades (includingnuclear factor kappa B (NF-κB), with convergent systems-level effects such as reorganization of the default mode network. Human studies report acute reductions in TNF-α with variable effects on IL-6 and CRP, consistent with an immunomodulatory profile. Pharmacokinetically, psilocybin shows properties advantageous in geriatric care: rapid onset, short half-life, and predominant phase-II glucuronidation, reducing interaction risk. Controlled studies demonstrate rapid antidepressant and anxiolytic effects in major depressive disorder, treatment-resistant depression, and existential distress, with emerging feasibility signals in neurodegeneration. Together, these findings support the hypothesis that a time-limited, mechanism-based intervention may improve mood and cognition while attenuating inflammation. This review integrates current evidence on psilocybin’s neuroimmune and pharmacokinetic mechanisms relevant to aging, outlining its potential role in inflammation-related disorders and highlighting the need for targeted studies in older adults, who remain underrepresented in psychedelic research. Full article

Background/Objectives: The emergence of antimicrobial resistance (AMR) within a One Health framework highlights the role of wildlife as environmental reservoirs. Because wild game is an increasingly important meat source, hygienic handling during evisceration is critical to prevent carcass contamination from internal tissues such as mesenteric lymph nodes (MLNs). This study aimed to investigate the occurrence and antibiotic resistance (AR) profiles of Escherichia coli isolated from the MLNs of hunted wild ungulates in western Romania to better understand microbiological hazards along the game meat supply chain. Methods: MLN samples were aseptically collected from 103 legally hunted wild boars (Sus scrofa, n = 78) and cervids (Capreolus capreolus and Cervus elaphus, n = 25) across two hunting grounds. E. coli isolation was performed utilizing selective Tryptone Bile X-Glucuronide agar. Subsequent biochemical identification and phenotypic antimicrobial susceptibility testing were conducted using the automated VITEK^® 2 system. Results: The overall E. coli isolation rate was 72.8% (75/103). Analyzed by host species, the bacterium was recovered from 79.4% of the sampled wild boars (62/78) and 52.0% of the cervids (13/25). Phenotypic resistance to at least one antibiotic agent was observed in 25.3% (19/75) of the isolates, most frequently against cephalosporins (cefalexin, 21.3%) and penicillins (ampicillin, 24.0%). Multidrug resistance (MDR) was identified in 20.0% (15/75) of the isolates. Conclusions: The detection of MDR E. coli phenotypes within the MLNs of free-ranging game indicates the penetration of clinically relevant resistance mechanisms into sylvatic environments. These findings underscore the potential risk of internal carcass contamination during field evisceration, highlighting the critical need for strict hygiene practices to ensure game meat safety. Full article

Background: Human diseases remain a major global health challenge, requiring effective therapeutic strategies. Traditional Chinese medicine (TCM) has been widely used in clinical settings. Many natural compounds, such as flavonoids from TCM, exhibit diverse pharmacological activities. Alpinetin and pinocembrin are structurally related flavonoids. Alpinetin is derived from Zingiberaceae plants, and pinocembrin is extracted from wild marjoram (origanum vulgare) or other natural sources. They possess a wide range of pharmacological activities or biological effects, including anti-inflammatory, anti-tumor, liver and kidney protection, cardiovascular protection, and antibacterial activities. Methods: The present comparative review was conducted in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, using four major databases (PubMed, EMBASE, Web of Science, and Cochrane Library), as well as CNKI without language restrictions. Results: Pharmacokinetic studies reveal distinct absorption, metabolism, and excretion profiles. Alpinetin and pinocembrin undergo glucuronidation and interact with cytochrome P450 enzymes and transporters. However, alpinetin has demonstrated approximately 1.5-fold higher plasma exposure and slower clearance compared to pinocembrin. Mechanistically, alpinetin exerted therapeutic effects through modulation of the NF-κB/MAPK, PI3K/Akt, and PPAR-γ signaling pathways, resulting in a 2- to 3-fold reduction in pro-inflammatory cytokines. In contrast, pinocembrin exerted protective activity through the inhibition of HMGB1/TLR4 signaling, regulation of endoplasmic reticulum stress, and activation of Nrf2/HO-1, leading to a 1.8-fold increase in antioxidant enzyme activity. The minimum inhibitory concentrations were reduced by 2- to 4-fold against Gram-positive bacteria compared to alpinetin. Conclusions: These findings highlight the pharmacological potential of alpinetin and pinocembrin as promising candidates for the development of novel anti-tumor, anti-inflammatory, liver and kidney protection, cardiovascular protection, and antibacterial agents. However, research on the pharmacological actions of alpinetin and pinocembrin is still in the preclinical stage. Further research is required to validate their efficacy in clinical settings, especially for translation to clinical studies. This is critical to translating these natural flavonoids into effective therapeutic agents while addressing the regulatory challenges and pathways associated with botanical drugs in human diseases. Full article

Background: Prenatal alcohol exposure (PAE) is a major preventable cause of neurodevelopmental impairment. Little is known about its impact on real-life child/adolescent academic outcomes, which are key functional indicators of long-term social participation. Methods: PAE was assessed in a sample of 156 participants via (1) meconium ethyl glucuronide (EtG) concentration (≥10 ng/g; EtG-positive, n = 36) and (2) third-trimester maternal self-report (SR-positive, n = 40). At primary school (6–10 years, t1) and secondary school ages (12–14 years, t2), exposed and non-exposed children were compared regarding primary school grades (t1) and secondary track placement (t2). Results: At t1, EtG-positive children exhibited lower school report grades by trend (n = 15, p = 0.068–0.085) with a moderate effect size (r = 0.36–0.39). SR-positive children demonstrated significantly lower school report grades (n = 15, p = 0.016) with strong effect (r = 0.56). At t2, both exposure groups had increased trend-significant odds of attending lower educational tracks (EtG-positive/-negative: n = 153, OR = 1.84, p = 0.059; SR-positive/-negative: n = 153, OR = 1.78, p = 0.066). Conclusions: PAE is associated with reduced real-life academic outcomes across development. School performance represents a sensitive functional outcome of neurodevelopmental vulnerability following PAE. Meconium EtG may contribute to postnatal identification of affected children. Full article

This study presents a multivariate assessment of the qualitative and quantitative composition of polyphenolic compounds across six Origanum species, including wild and commercial O. majorana, the three other species of section Majorana (O. dubium, O. syriacum and O. onites), and the more distantly related O. minutiflorum and O. vulgare. Methanolic extracts from 657 plants representing 59 populations were analysed by HPLC. A total of 122 constituents were consistently detected, 20 major peaks were selected for detailed evaluation, and eight key constituents were quantified using external standards. Wild O. majorana was characterised by high proportions of arbutin, apigenin 6,8-di-glucopyranoside, luteolin 7-glucuronide, apigenin 7-glucuronide, rosmarinic acid, salvianolic acid B, blumeatin, and two additional flavonoid-like constituents requiring further structural elucidation. Principal component analysis separated wild and commercial O. majorana and distinguished O. majorana, the three overlapping clusters of O. onites, O. dubium and O. syriacum, as well as O. minutiflorum and O. vulgare, reflecting marked interspecific differences in dominant compound classes. Origanum majorana and O. vulgare were richer in phenolic acids, whereas O. dubium, O. syriacum, and O. onites contained higher levels of flavonoid glycosides. Several genotypes accumulated exceptionally high concentrations of arbutin, apigenin 6,8-di-glucopyranoside, rosmarinic acid, or salvianolic acid B. These results establish a robust chemotaxonomic framework for distinguishing Origanum species and pinpoint high polyphenol genotypes as candidates for breeding, quality standardisation, and targeted follow-up studies on antioxidant and other bioactivities. Full article

Zearalenone (ZEN) is an estrogenic mycotoxin produced by Fusarium spp. and commonly found in cereals and feed materials. This study evaluated the ability of Metschnikowia pulcherrima KKP 1368 biomass to reduce extractable ZEN under controlled buffered pH conditions (pH 3.50 and 7.00) selected as simplified conditions relevant to the porcine gastrointestinal environment. ZEN was quantified by LC-MS/MS, whereas LC-MS-QTOF was used as a qualitative/semi-quantitative screening approach for tentatively assigned transformation-related features. In the presence of yeast biomass, extractable ZEN was already lower than in the corresponding controls at the first sampling point, indicating a rapid biomass-associated effect. After 12 h, reductions relative to the corresponding controls reached 63.0% at pH 3.50 (p < 0.0001) and 51.6% at pH 7.00 (p = 0.0001). ZEN remained stable in control samples, and the strain remained viable under both pH conditions throughout incubation. LC-MS-QTOF detected several tentatively assigned features consistent with zearalanone, zearalenone-14-glucuronide, and zearalenol O-glucoside; these assignments require confirmation with authentic standards. Overall, M. pulcherrima KKP 1368 reduced extractable ZEN in a simplified buffered in vitro system, probably through rapid adsorption/reduced extractability and possible biotransformation. Further studies using biomass fractions or inactivated biomass, mass-balance experiments, authentic standards, and toxicological assays are needed to clarify the relative contribution of adsorption and transformation and to assess the practical relevance of this approach. Full article

Background/Objectives: Women exposed to high psychosocial stress in pregnancy have higher risk of postpartum health conditions, but it still is unknown whether high pregnancy stress exposure alters the maternal metabolome at one-month postpartum. Methods: We analyzed data from 625 women participating in the PROGRESS study, a longitudinal pregnancy cohort. Women answered validated psychometric tests (i.e., EPDS, PSS and NLE) during the second or third trimester of pregnancy and provided serum samples at one-month postpartum for metabolomic assessment. Untargeted metabolomics were analyzed using chromatography high-resolution mass spectrometry (LC-HRMS). We used a metabolome-wide association study, using both traditional robust regressions and variance tests, to evaluate associations between pregnancy psychosocial stress and one-month postpartum serum metabolomics. Results: We found a few nominally significant associations between prenatal psychosocial stress scores and the maternal metabolome. However, these findings did not remain after adjusting for multiple testing, with the only exception of epiandrosterone glucuronide, a steroid hormone metabolite, and lithocholyltaurine, a lipid-like molecule. Conclusions: We did not find significant associations between prenatal psychosocial stress and postpartum serum metabolomic profiles, except for two metabolites showing suggestive associations warranting further investigation. Full article

Traditional edible plants such as quelites are an important component of the Mexican diet due to their nutritional and functional value; however, the effects of postharvest and culinary processing on their phytochemical composition remain poorly understood. This study evaluated the impact of oven-drying and freeze-drying, as well as thermal preparation (raw vs. boiled), on the proximal chemical composition, phenolic profile, and antioxidant capacity of leaves and inflorescences of Chenopodium berlandieri subsp. nuttalliae (huauzontle), using an integrated metabolomic approach. Proximal analysis showed that major macronutrients (protein, dietary fiber, lipids, and carbohydrates) were largely preserved across drying methods, whereas moisture and ash contents differed significantly among tissues and treatments (p < 0.05). Raw freeze-dried inflorescences exhibited the highest total phenolic content and antioxidant capacity. UPLC-DAD-ESI-QToF/MS enabled the identification and quantification of 26 phenolic compounds, predominantly glycosylated flavonols derived from quercetin, kaempferol, and isorhamnetin, while naringin was identified as the main flavanone glycoside present. Quercetin glucuronide was the most abundant compound, particularly in inflorescences. Multivariate analyses (principal component analysis [PCA], permutational multivariate analysis of variance [PERMANOVA], and partial least squares discriminant analysis [PLS-DA]) suggested that the drying method was a major source of variability, followed by thermal treatment and tissue type, although these patterns should be interpreted as indicative rather than conclusive. Overall, freeze-drying appeared to be the most effective method for preserving the phytochemical quality of huauzontle under the conditions evaluated, highlighting its potential as a valuable source of bioactive compounds within the genus Chenopodium. Full article

Background/Objectives: Synthetic stimulants represent the most prevalent subclass on the new psychoactive substances (NPSs) market. However, the toxicokinetic properties of M-ALPHA, a regioisomer of MDMA and N-methyl-cyclazodone a pemoline derivative, are not yet characterized. Methods: Therefore, this study investigated the metabolism of both NPSs in pooled liver S9 fraction and rat urine, characterized cytochrome P450 (CYP) kinetics and plasma protein binding (PPB), and assessed the CYP inhibition potential of M-ALPHA, using high-performance liquid chromatography coupled to high resolution tandem mass spectrometry (HPLC-HRMS/MS). Results: Four metabolites of M-ALPHA were detected including one phase I and three phase II metabolites, resulting from demethylenation followed by subsequent methylation or glucuronidation. For N-methyl-cyclazodone, one phase I metabolite formed via N-demethylation was identified. The primary enzymes involved in M-ALPHA metabolism were CYP2B6 and CYP2D6. Notably, M-ALPHA inhibited these enzymes to a strong or moderate extent, respectively. In contrast, the metabolism of N-methyl-cyclazodone was primarily mediated by CYP2A6. PPB studies indicated low-to-moderate binding for both compounds, suggesting that significant protein-binding interactions are unlikely. Conclusions: As M-ALPHA only formed metabolites that overlapped with those of MDMA, differing only by minor retention time shifts, reliable HPLC-HRMS/MS-based identification may be challenging in clinical and forensic toxicology settings as well as doping analysis. Furthermore, drug–drug interactions following polydrug use cannot be excluded for either NPS, particularly when co-ingested with other CYP substrates metabolized by the same isoforms. Full article

Background/Objectives: The most abundant flavonoid, quercetin, which is mostly found as glycosides, is widely distributed in plants. Quercetin is rapidly metabolized, having a short half-life in the blood circulation, and forms its conjugates by undergoing ring cleavage of the benzopyranone ring system. Despite its fast clearance in the body, quercetin was demonstrated to have clinically anti-inflammatory, cardioprotective, antidiabetic, and anti-obesity activities. This study aimed to determine whether quercetin itself or its metabolites are responsible for these activities. Methods: We performed molecular docking of 27 metabolites, including quercetin itself, against ten inflammation-related proteins in silico. We then conducted microscale thermophoresis (MST) of selected metabolites towards the NLRP3 inflammasome. Results: Overall, Phase II metabolites yielded better binding energies compared to the metabolites formed by degradation. MST results revealed that isorhamnetin, the 4-O-methylated metabolite of quercetin, gave the best results, with a binding affinity (K_D value) of 16.12 ± 5.16 µM, even better than quercetin itself, which has a binding affinity of 44.84 ± 4.21 µM. Glucuronide metabolites of quercetin (isorhamnetin 3-O-glucuronide, quercetin 7-O-glucuronide, and quercetin 3-O-glucuronide) were found to bind to the inflammasome protein with low binding affinities, whereas small degradation products (hippuric acid and 3,4-dihydroxytoluene) did not bind at all. Conclusions: These results suggest that Phase II metabolites, specifically isorhamnetin, may contribute more significantly to the biological activity of quercetin than the parent compound, however, degradation products appear inactive. Full article

Objectives: This study aims to identify the reactive metabolite of acetaminophen (AAP) and the cyanopyrrolidine metabolite of saxagliptin in human induced pluripotent stem cell-derived hepatic organoids (HHOs) and to compare them with human liver microsomes (HLMs) and plateable cryopreserved human hepatocytes (CHHs) to evaluate the feasibility of HHOs for reactive metabolite screening and metabolite profiling. Methods: AAP (50 μM) or sax-agliptin (50 μM) was incubated for 1 h at 37 °C in HLMs with or without NADPH-generating solution and 0.5 mM reduced glutathione (GSH). AAP (50 μM) was incubated for 24 h in HHOs and CHHs at 37 °C in a CO₂ incubator. AAP and saxagliptin metabolites in the reaction mixtures were analyzed using ultra-performance liquid chromatography coupled with tandem mass spectrometry. Results: N-acetyl-p-benzoquinone imine (NAPQI) was identified in the incubation mixture of HLMs with AAP, and its levels were reduced in the presence of GSH, accompanied by increased formation of AAP–GSH adduct. Incubation of AAP with HHOs for 24 h resulted in the formation of NAPQI, AAP–GSH, AAP–glucuronide, and AAP–sulfate. Moreover, CYP1A2 induction using omeprazole treatment increased the formation of AAP and AAP–GSH conjugate from phenacetin, reflecting enhanced CYP1A2 activity in both CHHs and HHOs. The findings indicate that HHOs are a suitable platform for reactive metabolites, such as NAPQI and AAP–GSH adducts, under chronic exposure and metabolic modulator intervention. Additionally, CHHs and HHOs exhibited similar saxagliptin metabolite profiles after incubation with saxagliptin and generated cysteine conjugates of saxagliptin and its hydroxylated metabolite. Conclusions: HHOs system can be used as an in vitro model for screening reactive metabolites, comparable to those obtained with CHHs. Full article

In this study, the pharmacokinetic (PK) characteristics of oleuropein present in the olive tree (Olea europaea) were determined. We developed and validated a highly sensitive ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS) method to quantify oleuropein and its aglycone derivative, thereby establishing their pharmacokinetic properties in vitro and in vivo. Quantification of oleuropein and oleuropein aglycone was performed using selected reaction monitoring (SRM) with heated electrospray ionization in negative ion mode, employing mass transitions of m/z 275.06 and 307.137 for the respective analytes, and methylparaben as the internal standard. The calibration curve for both exhibited a range from 1 ng/mL to 1000 ng/mL, utilizing a total of 10 calibrator standards. The method demonstrated superior sensitivity, precision, and reproducibility, facilitating accurate quantification of analytes over a wide concentration range in biological matrices. To develop a pharmacokinetic profile, C57BL/6 male mice were administered oleuropein at a dose of 100 mg/kg body weight via oral gavage, and plasma levels were examined by LC-MS/MS. Oleuropein pharmacokinetics were evaluated exclusively, as plasma levels of oleuropein aglycone remained below the limit of quantification throughout the 24 h sampling period. Mass analysis of plasma samples identified multiple glucuronidated and sulfated metabolites, establishing Phase II metabolism as the dominant pathway governing the systemic disposition of oleuropein. In addition, the metabolic stability of the compounds was also investigated in mouse liver microsomes and S9 fractions to define the in vivo stability of oleuropein and oleuropein aglycone. Full article

Diagnosis and monitoring of type 2 diabetes mellitus (T2DM) typically rely on invasive blood-based biomarkers. To explore non-invasive alternatives, this study examined tongue coating metabolites to identify metabolic signatures linked to diabetes progression. A case-control observational study categorized participants into control, prediabetes, and diabetes groups. Tongue coating samples were analyzed using liquid chromatography-mass spectrometry (LC-MS). Differential metabolites were correlated with clinical parameters, including HbA1c, BMI, and eGFR. Distinct metabolic profiles emerged across groups, with significant differences in five endogenous metabolites (phenylpyruvic acid, propionylcarnitine, pyridoxal 5′-phosphate, phenethylamine, phenethylamine glucuronide) and four amino acids (isoleucine, lysine, phenylalanine, tyrosine). Diabetic subjects showed elevated phenylpyruvic acid and phenethylamine, while propionylcarnitine, pyridoxal 5′-phosphate, and phenethylamine glucuronide were reduced. Phenethylamine was positively correlated with HbA1c; propionylcarnitine and phenethylamine glucuronide showed negative correlations with HbA1c and BMI. Detected total amino acids were inversely correlated with eGFR. Additionally, a diabetes index derived from these metabolic features also holds potential for discriminating disease states. These findings underscore the potential of tongue coating metabolites as a relatively non-invasive approach for evaluating T2DM states. The observed metabolic alterations provide valuable insights into diabetes-associated dysregulation, including protein glycation, obesity-related metabolic shifts, and renal impairment. Full article

Background/Objectives: Eudesmin is a tetrahydrofurofuranoid lignan known for its diverse pharmacological activities, including anti-tumor, anti-inflammatory, and neuroprotective effects. However, its metabolism has not been well characterized. Methods: This study examined the in vitro metabolism of eudesmin using human and mouse hepatocytes, human liver microsomes, and recombinant drug-metabolizing enzymes. Liquid chromatography–high-resolution mass spectrometry combined with ion identity molecular networking enabled the comprehensive visualization and annotation of eudesmin metabolites. Results: Eudesmin exhibited moderate metabolic stability in human and mouse hepatocytes, with half-lives of 181.0 min and 132.9 min, and intrinsic clearance values of 27.7 mL/min/kg and 154.0 mL/min/kg, respectively. Incubation of eudesmin with human hepatocytes resulted in the formation of 13 metabolites, including five phase I metabolites (M1–M5) and eight phase II conjugates. Phase I metabolism was dominated by O-demethylation of the 3,4-dimethoxyphenyl moieties, yielding mono-O-demethylated (M1 and M2) and di-O-demethylated metabolites (M3 and M4), as well as a hydroxylated metabolite (M5). Enzyme phenotyping, kinetic analyses, and chemical inhibition experiments identified cytochrome P450 2C9 (CYP2C9) as the major contributor to O-demethylation, with additional contributions from CYP2C19, CYP2C8, CYP3A4, and CYP3A5, whereas hydroxylation was mediated primarily by CYP3A4 and CYP3A5. The O-demethylated metabolites subsequently underwent phase II metabolism, forming glucuronide conjugates of M1–M4 and sulfate conjugates of M1–M3, including a disulfate of M3. Uridine 5′-diphospho-glucuronosyltransferase and sulfotransferase screening revealed the involvement of multiple conjugative enzymes, indicating extensive and distributed phase II metabolism. Specifically, di-O-demethylated metabolites and their conjugates were detected in human hepatocytes but not in mouse hepatocytes, suggesting that the sequential O-demethylation pathway is limited in mice. Conclusions: This study characterizes eudesmin metabolism, with CYP2C9-mediated O-demethylation and significant species differences between humans and mice, and provides a basis for its further pharmaceutical development. Full article