Search Results (343)

Persistently poor glioblastoma (GBM) survival necessitates better elucidation of tumor drug responses. After observing that low curcumin and all-trans retinoic acid (ATRA) doses stimulated cell proliferation and counteracted each other’s high-dose antiproliferative effects in U87 GBM cells, drug influences on cell growth, migration, and death and the antiproliferative interaction proteome were further studied. Cell proliferation and migration were assessed by xCELLigence Real-Time Cell Analysis (RTCA). Cell death was defined using flow cytometry. Drug interactions were determined with CompuSyn software (version 1.0). Liquid Chromatography–Tandem Mass Spectrometry (LC-MS/MS), High-Performance Liquid Chromatography (HPLC), and SequestHT software (version 1.4) were utilized for peptide generation and identification. ATRA at high doses inhibited cell growth and migration more efficiently. Curcumin was more proliferative and antagonistic against anti-growth effects at low doses. Migration inhibition and apoptosis occurred synergistically at the highest drug doses. ATRA influenced the proteome more remarkably, reducing Transforming Growth Factor Beta Induced (TGFBI), Phosphoglycerate Dehydrogenase (PHGDH), tenascin, and Sequestosome 1 (SQSTM1). These effects were alleviated by curcumin, except for SQSTM1. Uveal Autoantigen With Coiled-Coil Domains And Ankyrin Repeats (UACA) and Sad1 And UNC84 Domain Containing 2 (SUN2) were increased by ATRA and curcumin, and to a lesser extent by the combination. Hexokinase 2 (HXK2) was increased by curcumin and the combination. Heme Oxygenase 1 (HMOX1) was depleted by the combination, but not by the single agents. SQSTM1 and HMOX1 reductions may mediate anticancer synergism, while the remaining changes may indicate ongoing hormetic pathways not reflected in cell counts. Full article

Cariprazine (CAR) and lurasidone (LUR) are antipsychotic drugs used to treat schizophrenia. These drugs are relatively new in clinical practice, and there is a need to optimise and develop analytical methods for their determination in various biological matrices for biomedical analysis. To date, the detection of these drugs has been performed in serum and urine, but there are no methods for determining these drugs in saliva. In the first part of this study, various chromatographic systems were compared using high-performance liquid chromatography with diode array detection (HPLC-DAD) or coupled with quadrupole–linear ion trap tandem mass spectrometry (HPLC-QTRAP-MS/MS), taking into account the retention of tested compounds, system efficiency and peak symmetry. Next, a simple, rapid, and sensitive HPLC-QTRAP-MS/MS method has been developed for the determination of CAR and LUR in human serum, urine, and, for the first time, in saliva samples. Solid-phase extraction (SPE) was used for sample pre-treatment. Quantifications were carried out using a Polar RP column with a mobile phase consisting of acetonitrile and a formate buffer at pH 4.0 in gradient mode. The method was successfully applied for the determination of CAR and LUR in biological samples obtained from psychiatric patients. The findings suggest that saliva may be a non-invasive alternative for the quantification of free levels of investigated drugs, although further studies are required to clarify its relationship with plasma concentrations. Full article

Antimicrobial peptides (AMPs) are promising alternatives to antibiotics for combating multidrug-resistant bacteria, yet their practical application is hindered by the low content of natural AMPs and the high cost of chemical synthesis. In this study, we developed a high-efficiency heterologous expression system for the proline-rich cationic antimicrobial peptide Api137 in Saccharomyces cerevisiae CENPK2 by engineering the Ty retrotransposon system composed of multi-locus integration. Recombinant plasmids carrying Api137 encoding elements were constructed and integrated into the CENPK2 genome, generating the engineered strain CENPK2 + Ty1-2/2/3/4. The target fusion peptide (5.2 kDa) was successfully expressed and identified by Tris-tricine-SDS-PAGE and liquid chromatography–tandem mass spectrometry (LC-MS/MS). The quantification of Api137 from fermentation broth was applied by high-performance liquid chromatography (HPLC) which showed that the yield of tandem peptide in the fermentation supernatant reached 20.5 mg/L and the intracellular retention rate was 33.0%. Comparative analysis of MIC and MBC values revealed that the biologically synthesized Api137 exhibited slightly superior antibacterial activity relative to the chemically synthesized Api137. In vitro functional assays demonstrated that the fermentation supernatant of the engineered strain exhibited broad-spectrum antibacterial activity against five pathogenic bacteria, with a maximum antibacterial rate of 92.9% against Aeromonas veronii. Hemolysis assays and cytotoxicity tests confirmed that the fermentation supernatant exhibited neither hemolytic activity nor cytotoxicity. Moreover, the expression of Api137 did not impose a metabolic burden on the host. This study establishes a Ty transposon-mediated strategy for the high-level expression of Api137 in S. cerevisiae, which significantly demonstrates the antibacterial activity of Api137 while ensuring excellent biosafety. Full article

Antioxidant profiling of black locust (Robinia pseudoacacia L.) stem bark was performed by 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay off-line coupled to high-performance liquid chromatography–diode array detection–electrospray ionization mass spectrometry (DPPH–HPLC–DAD–ESI–QMS). The ethyl acetate, methanol, and n-hexane extracts exhibited similar chemical profiles for the antioxidant amphipathic phenolic compounds; however, the ethyl acetate extract was the richest in them, and n-hexane yielded several compounds in higher ratios. A total of 21 compounds displayed antioxidant activity and were further characterized by HPLC–high-resolution tandem MS (HRMS/MS). Nine compounds were identified as previously described antioxidants from the black locust stem bark, including 3-O-caffeoyl oleanolic acid (1), and the oleyl (4), octadecyl (6), gadoleyl (7), eicosanyl (9), (Z)-9-docosenyl (10), docosyl (12), tetracosyl (15), and hexacosyl (17) caffeates. Further ten caffeate and two ferulate esters were tentatively identified as 3-O-caffeoyl oleanolic, ursolic, or betulinic aldehyde (2), and hexadecyl (3), heptadecyl (5), hydroxy-hexacosyl (8), henicosyl (11), tetracosenyl (13), tricosyl (14), hexacosenyl (16), octacosenyl (18), and octacosyl (20) caffeates, and hexacosyl (19) and octacosyl (21) ferulates that were reported for the first time in R. pseudoacacia. Furthermore, this is the first report about the antioxidant activity of the previously undescribed compounds 8, 13, 16, and 18, as well as known compounds 11 and 14. Full article

Chili products are vulnerable to the addition of unauthorized substances, including Sudan dyes, Rhodamine B, Basic Orange 2, poppy-derived materials and improperly used processing chemicals. Their analysis is complicated by the high contents of lipids, carotenoids, capsaicinoids and other co-extracted matrix components in chili powder, chili oil, chili sauce and composite seasonings. This review critically evaluates conventional and emerging sample-preparation strategies, including solid-phase extraction; the quick, easy, cheap, effective, rugged and safe (QuEChERS) procedure; deep eutectic solvent (DES)-assisted extraction; enhanced matrix removal for lipids (EMR-Lipid); and molecularly imprinted sorbents. Laboratory methods based on high-performance liquid chromatography (HPLC), liquid chromatography–tandem mass spectrometry (LC–MS/MS) and gas chromatography–mass spectrometry (GC–MS) are compared with enzyme-linked immunosorbent assay (ELISA), surface-enhanced Raman spectroscopy (SERS), electrochemical sensors, miniature mass spectrometry and artificial intelligence-assisted hyperspectral imaging (AI–HSI). The comparison considers representative limits of detection and quantification, recovery, precision, sample-preparation burden, cost, portability, validation status and regulatory role. LC–MS/MS remains the preferred confirmatory platform for targeted multi-residue analysis, whereas rapid and portable methods are more appropriate for screening and sample triage. A three-tier workflow linking rapid screening, laboratory confirmation, and emerging-risk identification and traceability is proposed. Future priorities include standardized chili reference materials, open AI training and validation datasets, greener DES-based extraction and interlaboratory validation of field-deployable methods. Full article

Cervical cancer (CC) affects women worldwide, and more than 95% of cases are caused by persistent infection with high-risk human papillomavirus (HR-HPV), such as type 16, which promotes the progression of precancerous lesions to cancer. This study aimed to identify differentially expressed proteins (DEPs) in biopsies from patients with HPV16+ low-grade squamous intraepithelial lesions (LSILs) and from patients with HPV16+ squamous cell carcinoma (SCC) compared with those from HPV-negative normal cervical tissue (NCT HPV−) controls. The samples were analyzed by high-performance liquid chromatography–tandem mass spectrometry (HPLC-MS/MS) using a data-independent acquisition (DIA) approach. Data processing and differential protein expression analysis were performed with the DIA-NN software (Data-Independent Acquisition Neural Networks), followed by bioinformatics analyses, including Venn diagrams, pathway enrichment, functional interactome, The Cancer Genome Atlas (TCGA)-SCC data integration, and Western blot detection. In total, 1607 DEPs associated with cell adhesion and extracellular matrix proteins were identified in LSILs, whereas 1516 DEPs associated with catalytic and transport activities were identified in SCC; the proteins overexpressed in LSILs (332) were enriched in processes such as metabolism, immune response activation, and stress and cell death responses. In contrast, proteins overexpressed in SCC (205) were associated with the cell cycle, DNA damage, drug metabolism, proteasome degradation, methylation, and immune response. Interaction analyses highlighted proteins related to early proteins 1,5,6 and 7 (E1, E5, E6, and E7). In terms of the two DEPs, S100 calcium binding protein A10 (S100A10/p11) and thymidine phosphorylase (TYMP) were detected in patients with LSIL, HSIL, and SCC at the protein level, consistent with their higher transcript levels in public datasets. Given the small, exploratory cohort, these findings are hypothesis-generating, and validation in a larger, balanced, independent cohort is required. In conclusion, this study identified DEPs associated with the progression of premalignant lesions to SCC that may represent candidate biomarkers and therapeutic targets warranting further investigation. Full article

The food industry is interested in utilizing active compounds from agri-food waste to produce healthy products and promote sustainability and resource efficiency. This study aimed to conduct a phytochemical analysis of grape pomace derived from Vitis vinifera wine grape varieties Saperavi and Cabernet Sauvignon, as well as pomegranate (Punica granatum L.) peel of the Nar-Shirin variety, which are widely processed in South Kazakhstan. The extraction processes were simple and employed water and ethanol, two food-grade solvents widely used in green extraction due to their low toxicity and compatibility with food applications.Among the extraction methods tested, aqueous–alcoholic extraction demonstrated the highest efficiency in recovering total phenolic content (TPC), yielding 225.5 ± 1.46 mg GAE/g extract from pomegranate peel, 153.9 ± 1.25 mg GAE/g extract from Cabernet Sauvignon pomace, and 98.6 ± 0.83 mg GAE/g extract from Saperavi pomace. Phenolic profiling of the aqueous–alcoholic extract was performed using ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (HPLC-Q-TOF MS/MS). The total ion chromatograms revealed the presence of predominant phenolic compounds in all analyzed samples. Key identified constituents in grape pomace included kaempferol 3-O-β-D-xylofuranoside, kaempferol 3-O-β-glucopyranoside-7-O-α-rhamnopyranoside, quercetin-3-O-α-L-arabinoside, genistin, and megastigmane glycoside. In pomegranate peel, compounds such as myricetin, 5-(2Z,8Z)-pentadecadien-1-yl-1,3-benzenediol were identified for the first time. These findings highlight the rich phenolic composition of grape pomace and pomegranate peel’s extracts and support their potential as valuable sources of bioactive compounds for food, pharmaceutical, and industrial applications. Full article

Background/Objectives: Plant secondary metabolites remain an invaluable source of novel antibacterial phytochemicals in the fight against antibiotic resistance. The medicinal plant Solidago gigantea Ait. (giant goldenrod) is an invasive species in Europe and represents an abundant, yet largely underexplored reservoir of such bioactive compounds. The primary aim of this study was to perform a non-targeted, effect-directed screening, detection, bioassay-guided isolation, structure elucidation, and microbiological assessment of the antibacterial constituents present in the inflorescences of S. gigantea. Methods: Thin-layer chromatography coupled with direct bioautography (TLC–DB) assay using Bacillus subtilis was utilized for the non-targeted, effect-directed analysis of antibacterial components and the evaluation of in vitro antibacterial activity. Successive preparative flash column chromatography, semi-preparative reversed-phase high-performance liquid chromatography (RP-HPLC), and thin-layer chromatography–mass spectrometry (TLC–MS) were employed for the bioassay-guided fractionation and isolation. The structures of the isolated compounds were elucidated using one- and two-dimensional nuclear magnetic resonance (NMR) spectroscopy and high-resolution tandem mass spectrometry (HRMS/MS). The presence of known antibacterial compounds was established via reversed-phase ultra-high-performance liquid chromatography coupled with high-resolution electrospray ionization tandem mass spectrometry (RP-UHPLC–HR-ESI-MS/MS). Results: Two previously undescribed cis-clerodane diterpenoids, the isomeric solidagoic acid L (1) and solidagoic acid M (2), were isolated, identified, and characterized from the ethyl acetate extract of S. gigantea inflorescences. Both compounds exhibited in vitro antibacterial activity against the Gram-positive B. subtilis, confirmed via TLC–DB. In addition, 23 known compounds with antibacterial activity, including 17 clerodane diterpenes, four hydroxylated polyunsaturated fatty acids, and two unsaturated monoacylglycerols, were detected. All of these are reported for the first time in the inflorescences of this plant species. Conclusions: With further optimization, the isolated compounds may represent promising leads for antibacterial drug development. Our findings demonstrate the potential of non-targeted, bioassay-guided approaches for the discovery of novel plant-derived bioactive natural products. Full article

Background/Objectives: Tripterygium glycosides (TG) are used to treat inflammatory and autoimmune diseases, but their clinical application is limited by toxicity and the lack of process-responsive quality markers. This study examined whether roasting and dealkalization remodel the TG metabolite profile and alter the post-dose serum profile of circulating prototype constituents. Methods: Self-prepared TG, roasted TG (RTG), roasted–dealkalized TG (RDTG), and five marketed products were profiled by ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS). Seven representative compounds were quantified by validated high-performance liquid chromatography (HPLC). Rat serum after oral administration was analyzed to compare circulating prototype constituents. Results: We characterized 243 constituents in material samples and 63 circulating prototype constituents in serum. Roasting primarily reshapes the profiles of diterpenoids and triterpenoids. Celastrol was not detected in the RTG and RDTG material samples, nor in the corresponding single-time-point serum profiles under the current analytical conditions. In contrast, wilforlide A exhibited an increase in material samples. Dealkalization preferentially reduced alkaloid-related constituents, including wilforine in material samples and tripterygiumine T in serum. Conclusions: Integrated material profiling, targeted quantification, and serum prototype analysis identified candidate process-responsive markers for processed TG preparations. Because the serum study was based on relative signal intensities rather than full pharmacokinetics, these markers require further pharmacokinetic and toxicological validation. Full article

Mycotoxins are toxic secondary metabolites produced primarily by molds of the genera Aspergillus, Fusarium, and Penicillium. These widespread food and feed contaminants can cause significant risks to human and animal health. The aim of this study was to compare the performance of reference chromatographic methods (high-performance liquid chromatography coupled with either fluorescence detection (HPLC-FLD) or tandem mass spectrometry (HPLC-MS/MS)) with two commercially available rapid tests from two manufacturers. To that end, 90 randomly selected grain samples (barley n = 10, wheat n = 21, triticale n = 10, maize n = 49) collected in 2025 were analyzed for deoxynivalenol (DON), zearalenone (ZEN), ochratoxin A (OTA), and the sum of T-2 and HT-2 toxins. None of the samples exceeded the maximum levels established by the European Union (EU); however, widespread contamination with one or more mycotoxins was observed. Results showed that although rapid test strips offer advantages such as low cost, short analysis time, and operational simplicity, their considerably higher limits of detection and quantification values make them unsuitable for advanced laboratory analysis. Therefore, HPLC-FLD and HPLC-MS/MS remain the gold-standard methods for reliable, sensitive, and precise mycotoxin determination. Full article

Background: Hypertension-induced vascular injury involves endothelial dysfunction, inflammation, and oxidative stress, leading to vascular remodeling and cardiovascular complications. Flavin-containing monooxygenase 2 (FMO2) has been implicated in redox regulation, but its role in hypertensive vascular injury remains unclear. This study investigated whether KuJiang GanLuoYin (KJGLY) protects against hypertensive vascular injury and whether FMO2-associated Fat mass and obesity-associated protein (FTO)/N6-methyladenosine (m⁶A) signaling is involved. Methods: Spontaneously hypertensive rats (SHRs) were treated with KJGLY for eight weeks. Blood pressure, vascular remodeling, inflammation, oxidative stress, and global m⁶A RNA methylation were assessed. Integrated metabolomic and proteomic analyses were performed to identify treatment-associated molecular alterations and candidate proteins. AAV9-mediated FMO2 knockdown in SHRs and gain- and loss-of-function approaches in angiotensin II (Ang II)-stimulated human umbilical vein endothelial cells were used to examine the functional involvement of FMO2. Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS)-based chemical profiling and High-performance liquid chromatography–tandem mass spectrometry (HPLC–MS/MS) quantification were performed to characterize the major constituents of KJGLY. Results: KJGLY significantly reduced blood pressure and alleviated vascular remodeling in SHRs. Metabolomic and proteomic analyses revealed treatment-associated alterations in inflammatory and lipid metabolic pathways and identified FMO2 as a treatment-responsive candidate. KJGLY restored FMO2 expression, reduced FTO abundance and NF-κB activation, increased global m⁶A levels, and attenuated inflammatory and oxidative stress responses in hypertensive aortas. Conversely, AAV9-mediated FMO2 knockdown aggravated vascular injury, enhanced inflammation and oxidative stress, reduced global m⁶A levels, and increased NF-κB activation. Co-immunoprecipitation showed an association between FMO2 and FTO, and MeRIP-qPCR indicated that FMO2 manipulation altered m⁶A enrichment of VCAM-1 mRNA. In Ang II-stimulated endothelial cells, linarin, the most abundant quantified constituent of KJGLY, partially recapitulated the cellular effects of KJGLY, including restoration of FMO2/FTO-associated signaling and attenuation of inflammatory activation. Conclusions: These findings support a functional role for FMO2 in hypertensive vascular injury and suggest that FMO2-associated modulation of FTO/m⁶A signaling may contribute to the vascular protective effects of KJGLY. Linarin recapitulated key protective effects in vitro, although its in vivo contribution to the formula remains to be determined. Full article

Amaranthin is a major red-violet betacyanin of Amaranthaceae and an increasingly relevant natural pigment for food, cosmetic, nutraceutical, and biotechnological applications. This review integrates knowledge from over 100 studies, addressing amaranthin as a chemically defined betalain, distinguishing it from other scientific uses of the term, and evaluates its natural sources, analytical methods, extraction strategies, in vitro production systems, biosynthetic regulation, and biological activity. Cultivated Amaranthus species are among the richest plant sources, with total betacyanins of 46.1–199 mg/100 g fresh weight and amaranthin comprising, on average, 80.9% of the pigment fraction. Reliable identification and quantification rely on high-performance liquid chromatography coupled with a diode array detector (HPLC-DAD), liquid chromatography-tandem mass spectrometry (LC-MS/MS), and ultraviolet–visible (UV–Vis) spectrophotometry. Microwave- and ultrasound-assisted extraction can improve pigment recovery under optimized conditions, although its stability depends strongly on pH, temperature, solvent, time and storage parameters. While plant in vitro cultures, including callus, suspension, and shoot systems, have clarified biosynthetic regulation and offer controlled production platforms, engineered yeast systems have recently expanded production options, with Yarrowia lipolytica reaching 2.97 ± 0.029 g L⁻¹ amaranthin in fed-batch fermentation. Amaranthin-rich extracts and amaranthin-type pigments show antioxidantand anti-inflammatory potential, while antimicrobial and antiviral activities have mainly been reported for mixed betacyanin fractions; direct mechanistic, bioavailability, and in vivo evidence for purified amaranthin remains limited. Standardized analytical protocols, further investigation of stable high-yield sources, physicochemical stability assessment, and structure–activity studies are identified as priorities for advancing future application-oriented research on this multipotential pigment. Full article

Fifty-five batik textiles produced along coastal Java in the late 19th to early 20th century were analysed to study the red dyes and the cotton fabrics. Surface-Enhanced Raman Spectroscopy (SERS) classified the dyes into six groups and identified 70% of the samples as Morinda. High-Performance Liquid Chromatography coupled with a diode array detector and tandem Mass Spectrometry (HPLC-DAD-MS/MS) confirmed the SERS results and identified synthetic dyes in the remaining samples, which were used either alone or in mixtures with Morinda or indigo. Synthetic alizarin (C.I. 58000, Mordant Red 11) was the most frequently detected synthetic dye. Auramine O (C.I. 41000, Basic Yellow 2), fuchsin (C.I. 42510, Basic Violet 14), and rhodamine B (C.I. 45170, Basic Violet 10) were occasionally detected. The results also highlighted two possible types of Morinda and two variations of synthetic alizarin. The shades obtained from mixtures of natural and synthetic dyes were visually indistinguishable from those obtained with pure natural or synthetic dye, as confirmed by colourimetry. The variety of dyes and cotton fabrics shared across batik producers makes it challenging to attribute unsigned batiks to specific workshops. Nevertheless, this study demonstrated that synthetic dye uptake during this period was limited and experimental, with natural Morinda remaining the preferred choice despite the availability of European synthetic alternatives. Full article

Polymeric procyanidins (PPCs) constitute the major fraction of procyanidins, but they have poor bioactivity. The purpose of this study is to clarify the composition and content of PPCs from Vitis amurensis Rupr. (Vitaceae) seeds before and after depolymerization, thereby providing a theoretical basis for activity evaluation and application of proanthocyanidins (PCs). PPCs extracted from V. amurensis seeds were depolymerized by catechin-assisted sulfitation. The compositions and contents of PCs before and after depolymerization were qualitatively and quantitatively analyzed by ultra-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS), high-performance liquid chromatography (HPLC), and liquid chromatography–tandem mass spectrometry (LC-MS/MS). Results showed that twenty-eight components were identified (7 monomers, 13 dimers, 5 trimers, 1 tetramer and 2 unknowns). Before depolymerization, tetrameric and higher polymers dominated, accounting for 58.81% of the relative content. After depolymerization, these high-molecular-weight compounds declined to <1% or became undetectable, while monomers and dimers (with minor trimers) surged to 42.89%. Among them, the relative content of two monomers and three dimers, catechin, epicatechin gallate and procyanidin B1–B3, increased by 37.00, 3.75, 10.98, 3.72 and 9.74 times, respectively. In conclusion, the method utilizing catechin-assisted sulfitation effectively depolymerizes PPCs from V. amurensis seeds into oligomeric components such as monomers and dimers. Full article

The Ili River is a transboundary water body in Central Asia and a key region along the Belt and Road Initiative, yet antibiotic pollution in this river remains unexplored. This study investigated the occurrence, distribution, and ecological risks of 14 antibiotics in the Yining section of the Ili River and the effluent of the Huocheng Wastewater Treatment Plant (HWTP). We optimized and validated an analytical method using solid-phase extraction coupled with high-performance liquid chromatography-tandem–mass spectrometry (SPE-HPLC-MS/MS). The method achieved low detection limits (0.05–1.0 ng·L⁻¹) and good recoveries (70.0–108.3%). Results showed that HWTP removed 9.8–98.3% of detected antibiotics, with negative removal observed for several compounds due to desorption or metabolite reconversion. In the Yining river section, tetracyclines dominated (55.9% of total antibiotics), followed by fluoroquinolones (33.8%), macrolides (9.7%), and β-lactams (0.6%). Ecological risk assessment using the risk quotient method revealed that the combined risk (RQ_s) in the Yining section ranged from 1.54 to 2.74, indicating a high-risk level. Chlortetracycline exhibited the highest individual risk (RQ 0.92–0.97) and is proposed as a priority pollutant. This study provides the first baseline data on antibiotic pollution in a Central Asian transboundary river and underscores the need for international cooperation in water quality management within the Belt and Road framework. Full article