Search Results (15,980)

This paper describes a simple and sensitive method for determining the content of five bisphenols at the µg L⁻¹ level in tea infusion samples. The procedure uses a polypropylene hollow fiber filled with a deep eutectic solvent as the acceptor phase and 1-octanol as the supported liquid membrane, and the optimal conditions for the three-phase stir bar microextraction method were established as follows: a menthol–octanoic acid mixture (1:1 molar ratio) as the hollow-fiber filling, an extraction time of 1 h, and 80 µL of methanol for analyte desorption. The method demonstrated good linearity over the ranges of 1.5–30.0 µg L⁻¹ (BPF, BPA, BPAF, and BADGE) and 6.0–120.0 µg L⁻¹ (BPZ), with limits of detection between 0.28 and 1.01 µg L⁻¹, and the relative recovery values were satisfactory (99–120%) with acceptable precision (RSD < 17%). Thus, the method was successfully applied to quantitatively analyze twenty commercial tea infusions, in which BPF was detected at concentrations above the LOQ, and the greenness and overall applicability of the approach were confirmed using the AGREEprep and BAGI assessment tools. Full article

A comparative analysis of pentacyclic triterpenoids (ursolic acid and oleanolic acid) in the aerial parts of eleven Epilobium species was performed using high-performance liquid chromatography (HPLC) results. Samples for the analysis were collected from various regions in Ukraine, Poland and Lithuania. Oleanolic acid and ursolic acid were identified and quantified in nine of the species (E. angustifolium, E. montanum, E. collinum, E. roseum, E. palustre, E. tetragonum, E. obscurum, E. nervosum, and E. nutans). However, neither compound was detected in E. parviflorum or E. hirsutum from any collection site, indicating notable chemotaxonomic divergence within the genus. The quantity of ursolic acid in the analysed samples ranged from 20.27 ± 0.49 to 74.84 ± 2.24 mg/100 g dry weight, consistently exceeding that of oleanolic acid (2.03 ± 0.05 to 32.09 ± 0.73 mg/100 g). The highest total triterpenoid content was observed in E. tetragonum. These findings emphasise the importance of oleanolic and ursolic acids as auxiliary chemotaxonomic markers for Epilobium species. Given the well-documented antiproliferative and antiviral activities of these triterpenoids, the present results also suggest that several under-explored Epilobium species could be a promising source of bioactive compounds for further pharmacological research, particularly regarding prostate cancer and viral infections. Full article

Newborn screening for congenital adrenal hyperplasia (CAH) is effective in identifying patients with severe forms before a potentially lethal crisis, but has a relatively high false-positive rate. The aim of this study was to improve the national neonatal screening program in Sweden and the positive predictive value by implementing LC-MS/MS second-tier testing. A combination of two independent parameters, the steroid hormone ratio (androstenedione+17-hydroxyprogesterone)/cortisol and the concentration of 21-deoxycortisol and adjustment of cut-off levels resulted in an increase in the positive predictive value (PPV) from 14% to 84% for full-term infants. In total, the false-positive screening cases decreased by 88%. CYP21A2 genotyping was used to determine the severity of CAH in identified cases. We report on the stepwise approach that was used to optimize the cut-off levels for full-term and preterm infants in order not to miss any true cases in the process. Full article

Background: Experimental studies indicated that maternal choline and betaine status have the potential to alter fetal growth, but epidemiological data remain sparse. Objective: We aimed to investigate the association of maternal and cord blood choline and betaine concentrations with birthweight outcomes. Methods: This prospective cohort study involved 988 mother–infant dyads from Hebei and Shandong provinces. Plasma concentrations of choline and betaine in maternal late pregnancy and cord blood were quantified using ultra-performance liquid chromatography–mass spectrometry. Multivariable linear or logistic regression was performed to examine their association with continuous or binary birthweight outcomes. Results: Maternal plasma choline and betaine concentrations in late pregnancy (median [interquartile range]; 12.34 [10.13, 14.78] and 14.99 [12.01, 18.36] μmol/L) were significantly lower than those in cord blood (29.98 [24.74, 35.93] and 31.14 [25.56, 37.28] μmol/L). Each 1 μmol/L increase of late-pregnancy and cord blood betaine concentrations were associated with 9.87 g (95% confidence interval [CI]: −16.08, −3.66 g) and 5.29 g (95% CI: −8.52, −2.06 g) lower birthweight, respectively. Compared with the lowest quintile, the highest quintiles of late-pregnancy and cord blood betaine concentrations were associated with lower risks of large-for-gestational-age (adjusted odds ratios [ORs] = 0.47 [95% CI: 0.24, 0.90] and 0.31 [95% CI: 0.17, 0.56]) and macrosomia (adjusted ORs = 0.12 [95% CI: 0.03, 0.43] and 0.15 [95% CI: 0.05, 0.40]). These associations, particularly for cord blood, persisted and appeared more pronounced in pregnancies with maternal overweight/obesity or gestational diabetes mellitus (GDM), but the interaction effect did not reach statistical significance. No significant associations were observed for choline in any periods. Conclusions: Higher plasma concentrations of betaine in maternal late-pregnancy and cord blood were associated with lower birthweight. These findings emphasize the importance of sufficient betaine status during pregnancy, especially among mothers with obesity or GDM. Full article

We evaluated the antimicrobial and antioxidant capabilities of a bacteriocin purified from a recently identified marine Lactococcus lactis (L. lactis) NAN6399 strain, a lactic acid bacterium recovered from Mediterranean coastal waters near Alexandria, Egypt, and identified by combined API 50 CHL phenotypic profiling and 16S rRNA gene sequencing. Bacteriocin purification was achieved by sequential ammonium sulfate precipitation and reverse-phase high-performance liquid chromatography (RP-HPLC). The purified bioactive fraction had an approximate molecular weight of 20 kDa by SDS-PAGE and a 106-amino-acid N-terminal sequence that, upon BLAST alignment, returned 98.1% overall identity to the Lactococcin 972 family bacteriocin AAK06118.1 from L. lactis IL1403, with divergence confined exclusively to the terminal two C-terminal residues. This sequence is structurally and functionally distinct from canonical Lcn972 (L. lactis IPLA 972): the two peptides share an identical 25-residue signal peptide but diverge entirely in their mature bioactive domains, which exhibit only 9.1% sequence identity. Canonical Lcn972 operates through Lipid II-mediated septum disruption and inhibits only Lactococcus species; the NAN6399 peptide, correctly designated as a novel member of the Lcn972-like peptide family, demonstrated broad-spectrum antimicrobial efficacy against multiple indicator organisms (Staphylococcus aureus, Salmonella typhimurium, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Enterococcus faecalis), producing inhibition zones of up to 30 mm and minimum inhibitory concentration (MIC) values as low as 1.25 μg/mL against S. aureus. Antioxidant capacity was assessed using the DPPH radical scavenging assay, with the purified preparation achieving 73.14 ± 0.34% inhibition. Collectively, these data establish L. lactis NAN6399 as the producer of a bifunctional Lcn972-family bacteriocin with both antimicrobial and antioxidant potential, provide the first experimental characterization of the antimicrobial activity of this Lcn972-family branch, and highlight marine LAB as a productive reservoir for novel bioactive peptide discovery. Full article

Background: Four genotypes of human bocaviruses (HBoVs) have been identified, with only HBoV1 being detected in respiratory specimens, and with HBoV2 being the predominant human bocavirus in fecal specimens, which implies different tissue tropisms for HBoV1 and HBoV2. It is vital to determine the factors that influence the tissue tropisms. Methods: The major capsid proteins VP3 of HBoV1 and HBoV2 were expressed in eukaryotic cells. Then co-immunoprecipitation (Co-IP) and liquid chromatography–tandem mass spectrometry (LC-MS/MS) (IP-MS) was employed, along with Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses, to screen host proteins interacting with VP3 of different genotypes. Subsequently, in vitro pull-down assays were conducted to verify the direct virus–host interaction proteins with VP3. Furthermore, molecular docking was performed to predict the interaction interfaces between viral and host proteins. Results: Through IP-MS and enrichment analyses, 50 host proteins that displayed ≥10-fold differential binding affinities between HBoV1 VP3 and HBoV2 VP3 were identified. Among these, seven were considered as high-confidence candidate interactors. Notably, SPLUNC1 and VAMP8 showed predominant expression in respiratory and intestinal tissues, respectively. Subsequent in vitro pull-down assays confirmed that SPLUNC1 specifically bound to HBoV1 VP3, whereas VAMP8 specifically interacted with HBoV2 VP3. Molecular docking analysis further revealed that the binding between SPLUNC1 with HBoV1 VP3, as well as VAMP8 with HBoV2 VP3, was stabilized by extensive hydrophobic interfaces along with specific hydrogen bonds. Conclusions: The specific interactions of host proteins SPLUNC1 with HBoV1 VP3 and VAMP8 with HBoV2 VP3, respectively, provided fundamental evidence that the distinct tissue tropisms of HBoVs may be governed by specific host factors. Full article

Ertugliflozin is a sodium-glucose co-transporter-2 inhibitor that has demonstrated promise as a treatment for hyperinsulinaemia in horses. Despite the frequent use of ertugliflozin in equine clinical practice, the pharmacokinetics of this drug in horses has not been established. The aim of the present study was to determine the pharmacokinetics of one supratherapeutic dose (0.25 mg/kg) of ertugliflozin in eight horses. Horses were defined as being healthy by physical examination, haematological, blood biochemical and oral sugar test (OST) results. Plasma concentrations of ertugliflozin were quantified using high-performance liquid chromatography–tandem mass spectrometry 0, 0.25, 0.5, 0.75, 1, 2, 3, 4, 6, 8, 10, 14, 18, 24, 30, 36, 48, 60, 72, 96, and 120 h after drug administration enterally. Non-compartmental analysis led to determination of key pharmacokinetic variables, including mean ± SD time to maximum concentration (T_max) of 0.91 ± 0.13 h, maximum measured concentration (C_max) of 267.52 ± 25.37 ng/mL, terminal elimination half-life (T_1/2) of 17.65 ± 3.15 h and apparent oral clearance (CL/F) of 106.95 ± 27.53 mL/h/kg. No clinical signs of adverse effects or blood biochemical abnormalities occurred after drug administration. The results of this study suggest that a single supratherapeutic dose of ertugliflozin in healthy horses is safe. The pharmacokinetics of enterally administered ertugliflozin in horses are similar to pharmacokinetics of the drug in humans and the long T_1/2 makes ertugliflozin suitable for once daily dosing in horses. It is proposed that a starting dose for ertugliflozin in horses be in the range 0.05–0.1 mg/kg. Further pharmacokinetic studies are required to optimise the dose regimen for treating horses with hyperinsulinaemia. Full article

Recently, Quzhou Aurantii Fructus Extract (QAFE) was reported to exert anti-inflammatory effects on different types of respiratory diseases; however, it is yet to be determined whether it is effective in patients with idiopathic pulmonary fibrosis (IPF). The purpose of this study is to explore the capacity of QAFE to fight fibrotic disease, in particular how it works in relation to the regulation of the nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase 1 (HO-1) pathway. QAFE was made with Quzhou Fructus Aurantii (QAF), and the content of four flavonoids in the samples prepared was analyzed by high-performance liquid chromatography (HPLC). The therapeutic effect of QAFE was experimented by establishing IPF models in mice and in cells. Identification of the mechanism of QAFE in IPF through knockout or knockdown of the Nrf2 gene. The experiments suggest that QAFE has the potential to prevent IPF-induced inflammation, collagen deposition, oxidative stress, and apoptosis of cells. Silencing Nrf2 by knockdown or knockout is enough to prevent the capacity of QAFE to inhibit the process of inflammation, oxidative stress, and collagen deposition to cause more serious lung injury and HO-1 expression downregulation. QAFE is a potential new antifibrotic drug in IPF with an effect on the Nrf2/HO-1 pathway that reduces inflammation and oxidative stress. Full article

Background: Phospholipids are essential components of bacterial membranes and play central roles in membrane integrity and adaptation to antibiotic stress. However, confident annotation of phospholipid molecular species remains challenging due to the complexity of the lipidome and the limited structural constraints in conventional lipidomics workflows. Methods: Here, we present a bacterial phospholipidomic framework that integrates orthogonal structural evidence to achieve high-confidence and traceable annotation. Thin-layer chromatography (TLC) provides phospholipid headgroup assignment, gas chromatography–mass spectrometry (GC–MS) defines the acyl-chain pool, and Paternò–Büchi derivatization enables C=C localization, collectively restricting the structural search space prior to liquid chromatography–tandem mass spectrometry (LC–MS/MS) analysis. A rule-based ion prediction library further standardizes diagnostic ion assignment and reduces annotation ambiguity. Results: Applying this platform, we found Escherichia coli in the stationary phase remodeled the membrane phospholipids, with cardiolipin (CL) increasing from ~5% to ~10% and cyclopropane-containing phospholipid species rising to ~75%. Similar remodeling patterns are observed under diverse antibiotic exposures at sub-inhibitory concentrations, consistent with convergence toward a tolerance-associated membrane state. Extension of the framework to Enterococcus faecium supports proof-of-concept application in an additional Gram-positive model, with vancomycin-resistant strains exhibiting pronounced phosphatidylglycerol (PG) enrichment and reduced CL. Conclusions: Our work provides a scalable and reproducible strategy for bacterial phospholipid annotation, enabling molecular-species-resolved investigation of membrane adaptation and offering a framework for future exploration of lipid homeostasis pathways as potential antimicrobial targets. Full article

Background: Predictive modeling of vaccine stability is an essential tool for accelerating development and ensuring product quality, particularly when long-term data are limited. To ensure high-quality input for accurate stability predictions, it is often necessary to allocate substantial analytical resources. Methods: This study demonstrates that integrating high-throughput screening (HTS) techniques such as ultraviolet–visible spectrophotometry (UV-VIS), intrinsic fluorescence and dynamic light scattering (DLS) with Advanced Kinetics Modeling (AKM) results in a synergistic approach, facilitating the development of robust predictive stability models. Results: Here, we applied AKM to a glycoconjugate vaccine targeting extra-intestinal pathogenic Escherichia coli (ExPEC). The aggregation processes observed via size-exclusion high-performance liquid chromatography (SE-HPLC), DLS, turbidity by UV-Vis absorbance and local changes in tryptophan microenvironment captured by intrinsic fluorescence were effectively described by the developed kinetic models. Conclusions: Using accelerated stability data across multiple temperatures, AKM successfully described key degradation pathways. Furthermore, the HTS assay results showed strong correlation with SE-HPLC data, indicating that these assays provide an efficient alternative requiring minimal analytical resources, material, and time. Full article

Objectives: Concentrated pills, as a modernization and upgrade of traditional pills, have achieved significant advancements in dosage form. However, their extended disintegration and dispersion times have become a major limitation to their therapeutic efficacy. Therefore, an in-depth study and explanation of the dissolution mechanism of concentrated pills, along with the development of processing technology to control dissolution time, has emerged as a critical bottleneck in improving the quality of concentrated pills. Methods: In this study, the Liuwei Dihuang (LWDH) concentrated pill, derived from the classical Liuwei Dihuang pill, was selected as a representative model. Two drying methods—hot-air drying and hot air–microwave combined drying—were comparatively investigated to evaluate their effects on dissolution time. The dissolution behavior was elucidated by analyzing water migration during the dissolution process, as well as the textural properties and internal structural characteristics of the pills using Low-Field Nuclear Magnetic Resonance (LF-NMR), Micro-Computed Tomography (Micro-CT), texture analysis, and other modern techniques. Results: The results indicated that: (i) The rate of water absorption during the dissolution process of the LWDH pill was influenced by the number and size of the internal pores. (ii) Hot air–microwave combined drying results in more pores and faster dissolution. (iii) High-Performance Liquid Chromatography (HPLC) fingerprints showed no significant differences in the active ingredients between the samples. Conclusions: The drying method significantly affected the internal structure of the pills, suggesting that controlling the drying process could address the prolonged dissolution time of concentrated pills. Full article

Over the past decade, veterinary forensic toxicology has gained increasing relevance due to the recognition of animal poisoning as a criminal, environmental, and public health concern. This review provides an updated overview on current perspectives in veterinary forensic toxicology, focusing on common toxicological scenarios, analytical innovations, and interpretative challenges. Intentional poisoning of companion animals, wildlife intoxication, and environmental contamination are discussed within a One Health framework. Mass spectrometry-based techniques, including targeted gas chromatography–tandem mass spectrometry (GC-MS/MS), liquid chromatography–tandem mass spectrometry (LC–MS/MS) and high-resolution mass spectrometry (HRMS) for non-targeted screening, and emerging ambient mass spectrometry approaches are highlighted as pivotal tools in modern forensic investigations. Major limitations related to post-mortem changes, species-specific toxicokinetic, and the lack of harmonized interpretative criteria are critically examined. The review also identified future priorities such as methodological standardization, expansion of toxicokinetic databases across species, detection of emerging contaminants, and strengthened international cooperation. Overall, veterinary forensic toxicology is an emerging discipline at the intersection of animal welfare, environmental conservation, and public health providing essential evidence for legal, regulatory, and investigative purposes. Full article

Background/Objectives: Understanding the causes of bull subfertility and developing reliable diagnostic tools are critical to reducing economic losses caused by reproductive failure in beef cattle. Metabolomic analysis of sperm from bulls with diverging field fertility may provide insights on sperm metabolism that are associated with fertility. The objective was to determine metabolomic differences in sperm from bulls with differing field fertility. Methods: Angus bulls (n = 15) were classified based on a composite field fertility index (CFI). Frozen–thawed semen straws (n = 10 per bull) underwent a Percoll gradient sperm purification process. Metabolomic analysis was performed through ultra-high performance liquid chromatography coupled high resolution mass spectrometry at the University of Tennessee Biological and Small Molecule Mass Spectrometry Core. The general linear model (GLM) procedure of Statistical Analysis System (SAS) was used to evaluate linear and quadratic relationships between metabolites and CFI. Furthermore, the MIXED procedure was used to determine differences in metabolite abundance between the four highest and lowest fertility bulls. Significance was determined when p ≤ 0.05 and tendency was declared when p ≤ 0.10. Results: A total of 75 metabolites were detected. Quadratic relationships with fertility were observed for kynurenine, xanthine, and ophthalmate. Tricarballylic acid and creatinine showed a negative linear relationship with fertility. When differences in metabolite abundance were assessed between the four highest and lowest fertility bulls, N-acetylglutamate and N-acetylglutamine had greater abundance in low fertility bulls. Conclusions: Metabolites kynurenine, xanthine, ophthalmate, tricarballylic acid, and creatinine are potential fertility markers to identify subfertile bulls from a breeding population. These metabolites have promising future implications in the diagnosis and treatment of beef bull subfertility. Full article

The Cape cobra (Naja nivea), one of Africa’s most lethal snakes, can cause rapid, life-threatening paralysis. However, the impact of this venom on platelet function and blood coagulation remains poorly understood. To address this gap, we investigated the enzymatic profiles and the impacts of N. nivea venom on multiple aspects of haemostasis using human whole blood. Our results illustrate that Cape cobra venom significantly increases clotting time in rotational thromboelastometry without affecting other coagulation parameters. This venom significantly inhibits platelet aggregation and activation yet does not exert cytotoxic effects on platelets. The venom was subsequently fractionated using reverse-phase high-performance liquid chromatography, and the most potent purified fraction was identified as a cytotoxin (three-finger toxin) through mass spectrometry. This purified fraction showed an inhibitory effect on platelet activity. These findings highlight that N. nivea venom can induce haemotoxicity in addition to neurotoxicity. Moreover, three-finger toxins may be promising candidates for bioprospecting to develop novel antithrombotic agents. Full article

Pyrolysis liquid (PL) derived from biomass pyrolysis exhibits biopesticidal properties and represents a promising value-added product within the sustainable circular economy framework. However, knowledge about the antimicrobial potential of PLs produced from walnut residue at different pyrolysis temperatures remains limited. We investigated the chemical composition and antimicrobial activity of PLs obtained from agricultural walnut residue (Juglans regia L.) against selected plant pathogenic bacteria and fungi. PLs were produced at four temperature ranges: 200–300 °C (W-1), 300–400 °C (W-2), 400–500 °C (W-3), and 500–600 °C (W-4). Chemical characterization was performed using Gas chromatography–mass spectrometry (GC-MS), High-performance liquid chromatography (HPLC), and Inductively coupled plasma optical emission spectrometry (ICP-OES), with determination of total phenolic and flavonoid contents. Pyrolysis temperature significantly influenced the chemical profile and bioactive compound content of the PLs, with W-4 showing the highest total phenolic and flavonoid levels. Heavy metal analysis indicated minimal contamination in all samples. Antibacterial activity was observed in stock solutions, whereas diluted applications showed limited effects. The W-4 fraction showed the strongest antibacterial activity and exhibited MIC values of 12.50 µL/mL against Clavibacter michiganensis subsp. michiganensis, Xanthomonas euvesicatoria, and Pseudomonas syringae pv. syringae, and 25.00 µL/mL against Erwinia amylovora. Antifungal activity differed markedly across temperature ranges, with W-3 and W-4 displaying superior activity against Fusarium oxysporum and Verticillium dahliae, achieving complete mycelial growth inhibition at 5%, compared to 10% for W-2 and 20% for W-1. Positive controls confirmed assay validity (ciprofloxacin for antibacterial assays and cycloheximide for antifungal assays), whereas negative controls showed no inhibitory effect. Overall, higher pyrolysis temperatures, particularly 400–600 °C, enhanced the antimicrobial potential of walnut residue-derived PLs, supporting their possible use as bio-based antifungal agents for sustainable crop protection. Full article