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Microbial libraries remain an important resource for natural product discovery; however, constructing taxonomically and chemically diverse collections remains a challenge. Advances in dereplication strategies, including molecular networking, have reduced the rediscovery of known bioactive molecules and facilitated the identification of novel chemical scaffolds, but these approaches are typically applied after library construction or to existing repositories. Furthermore, many dereplication workflows require scaled fermentation and extraction, increasing the time needed to assess a microbe's metabolite profile. Here, we integrate matrix-assisted laser desorption/ionization tandem mass spectrometry (MALDI-MS/MS) into the bioinformatics platform IDBac, enabling streamlined characterization of microbial taxonomic identity, metabolite production potential, and preliminary metabolite annotation through GNPS2 molecular networking. This miniaturized high-content workflow facilitates strain prioritization by providing metabolite annotations directly from single microbial colonies prior to scale-up and extraction. Application of this approach to marine actinomycetes enabled the annotation of lavanducyanin and multiple napyradiomycin analogs. Subsequent investigation led to the discovery of napyradiomycin B8 from marine Streptomyces sp. CNZ-289, which was confirmed by 1D and 2D NMR spectroscopy and MALDI-MS/MS. Expanding this workflow to an untargeted analysis of 25 commensal marine vertebrate-derived bacterial isolates resulted in the annotation of several known bioactive natural products, including surugamides, antimycins, desferrioxamine siderophores, and the isolation and elucidation of harmane derivatives using NMR.
This study aimed to evaluate the value of nucleic acid-based matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) in identifying mycobacterial species and detecting drug resistance in patients with retreatment pulmonary tuberculosis (TB). The clinical data of 100 patients with retreatment pulmonary TB treated at Wenzhou Central Hospital between 1 June 2022 and 31 December 2023 were retrospectively collected and analyzed. Respiratory secretions were examined using BACTEC MGIT 960 Rapid Liquid Culture (MGIT 960 culture), drug-sensitive gene chip strain identification and nucleic acid-based MALDI-TOF MS assay (MassARRAY® system). Unlike traditional protein-based MALDI-TOF MS, this nucleic acid-based approach detects specific gene mutations associated with drug resistance. The results of the MGIT 960 culture and drug resistance testing served as the benchmark for evaluating the sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of the MALDI-TOF MS assay. For Mycobacterium tuberculosis (MTB) detection, compared with the MGIT 960 culture, the nucleic acid-based MALDI-TOF MS assay demonstrated a sensitivity of 97.01% (65/67), specificity of 63.64% (21/33), PPV of 84.42% (65/77) and NPV of 91.30% (21/23), with a concordance of 86.00% and a kappa value of 0.619. The difference in detection results between the two methods was statistically significant (χ2 = 15.625, p < 0.001). For drug-resistance detection in the 34 MTB-positive cases with both methods, concordance with MGIT 960 was 97% for rifampicin (97% sensitivity, 100% specificity) and 88% for isoniazid (86% sensitivity, 100% specificity), with overall κ = 0.83 across first- and second-line drugs. The nucleic acid-based MALDI-TOF MS assay demonstrates promising performance in rapidly identifying MTB and detecting drug resistance, serving as a valuable complementary tool for rapid clinical diagnosis and personalized medication guidance in patients with retreatment TB. The detection of non-tuberculous mycobacteria requires further validation with larger sample sizes.
The majority of chemical signals detected in public metabolomics repositories remain structurally undefined. Large language models (LLMs) are probabilistic systems whose capacity to generate outputs beyond their training data, which can cause hallucinations, makes them also potentially suited to hypothesize structures for molecules that have never been described. We aimed to build a system that could harness this LLM generative capacity combined with domain specific tools/framework to constrain hallucination and produce validated discoveries. We developed a GNPS2 agentic AI system that interprets LC-MS/MS data by integrating spectral alignment, molecular formula inference, rule-based structural enumeration, machine learning-based spectrum prediction, and translates natural language hypotheses from domain experts into dynamically generated analytical workflows. We demonstrate the annotation of unknown drug metabolites from public data guided by chemical hypotheses. The agent predicted, and we experimentally confirmed, a phosphorylated hydroxyzine, an acetaminophen-p-coumaric acid ester, and identified two new oxidative ibuprofen-carnitine conjugates from public repositories. These results demonstrate that LLM-driven agentic reasoning, when combined with domain expertise, can indeed generate experimentally testable structural hypotheses for previously uncharacterized metabolites leveraging pan repository data.
Non-canonical amino acids (ncAAs) are valuable tools in chemical biology and biochemistry for labeling, probing, and tracking biomolecules. ncAAs that can be recombinantly incorporated using native E. coli machinery are particularly useful because they allow for global protein incorporation and avoid complex genetic code expansion. Here, we demonstrate successful incorporation of a methionine analog, L -cyanohomoalanine (Cha), by the methionyl-tRNA synthetase of E. coli into mutant superfolder GFP (sfGFP) expressed in methionine auxotroph bacterial cultures. We compare to methionine auxotroph bacterial cultures supplemented with L- methionine (Met) or L -azidohomoalanine (Aha). In control prototrophic E. coli , bacterial growth rates are inhibited with high concentrations of Aha but not Cha. However, less sfGFP is produced in auxotrophic cells supplemented with Cha compared to Aha and Met. Thus, while Cha is non-toxic to E. coli it is incorporated less efficiently into proteins than Aha or Met. Mass spectrometry confirmed that N-terminal Cha, Aha, and Met are cleaved, as expected for the sfGFP mutants. Other sites of Cha and Aha incorporation were confirmed by mass spectrometry, with labeling efficiency varying by position. Thermal melts of purified sfGFPs demonstrate that Cha and Aha labeling does not significantly perturb the protein stability. In the future, Cha may be useful for proteome labeling by wild-type methionyl-tRNA synthetase and could be implemented in metabolic pulse-labeling of newly synthesized proteins with other methionine analogs. Additionally, the nitrile moiety of Cha may be used to perform reactions orthogonal to azide/alkyne click chemistry or could serve as a vibrational reporter of the environment.
Elicitation is an important strategy for producing plant secondary metabolites, which are responsible for a plant's highly dynamic chemical defense against environmental stressors such as UV light, predators, and pathogens. Elicitation strategies have recently employed nanoparticles as carriers to effectively deliver the elicitation agent through the cell membrane. This study examines the elicitation of secondary metabolites in Lobelia cardinalis hairy root cultures (HRCs) in response to the plant hormone jasmonic acid (JA) and a JA-loaded nanoparticle carrier (mesoporous silica nanoparticles (MSNPs)). Ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS) with high resolution mass spectrometry identified 12 m/z features only upregulated in the treatments with JA (free JA, JA-loaded MSNPs, and physical mixtures of JA and MSNPs) relative to a control experiment (without JA) or exposure to MSNPs alone. Putative identifications for these compounds elicited by JA included sesquiterpenoids (rishitin), monolignols (elemicin), and coumarins (7-hydroxycoumarin derivatives), respectively, and roughly correlated with the levels of JA that was measured in the respective HRCs. Signature m/z analytes associated with exposure to MNSPs were identified across treatment of bare MNSPs, JA-loaded MSNPs, and physical mixtures of JA and MSNPs and are consistent with a general stress response. The study demonstrates the ability to differentiate the elicitation effects of a biotic elicitor and its delivery system in untargeted elicitation studies, with a goal of designing nanocarrier systems for effective production of secondary metabolites.
Brains decompose rapidly after death yet frequently survive in the archeological record, including >1300 cases of waterlogged, oxygen-poor graves in which the brain is the only preserved soft tissue amongst otherwise skeletonized remains. To address this paradox, we decayed mouse carcasses for six months in four burial regimes varying in water and oxygen availability, characterized the brain proteome by high-resolution liquid chromatography-tandem mass spectrometry using parallel data-dependent and -independent acquisition workflows at six post-mortem intervals, and modeled >1.26 million peptide-specific decay trajectories to identify decay-prone and -resistant sequences. Oxygen availability exerted the main control on molecular fate: oxic burials produced widespread protein loss, whereas wet, hypoxic conditions favored retention of a distinctive subset of decay-resistant peptides. These surviving sequences were structurally ordered, enriched in redox-active residues and in regions that bind metals and lipids, and bore modification patterns consistent with radical-mediated oxidative cross-linking rather than fragmentation. By linking intrinsic tissue chemistry and environmental context, our results move brain preservation from anomaly to expectation: resolving why brains outlast other soft tissues in waterlogged, oxygen-poor burials, and revealing that the molecular signatures of post-mortem peptide persistence closely mirror those of pathological protein stabilization in brain aging and neurodegeneration.
Jusvinza is an immunomodulatory pharmaceutical drug developed for the treatment of rheumatoid arthritis. Its active ingredient is an altered peptide ligand-derived from a T-cell epitope of the human 60-kDa heat shock protein-referred to as CIGB-814. Jusvinza has also proven effective in reducing hyperinflammation in COVID-19. However, molecular targets of CIGB-814 in patients remain undefined, and this study is aimed at identifying specific plasma proteins that interact with CIGB-814. Affinity chromatography and mass spectrometry analysis revealed apolipoprotein A-I as a CIGB-814-binding partner. Interestingly, no apolipoprotein A-I bands were detected in the affinity matrix of the wild-type peptide (designated as E18-3). In this context, it was relevant to analyze how the substitution of Asp18 with Leu-used to design CIGB-814 from the wild-type peptide-confers its ability to bind apolipoprotein A-I. Docking studies predicted the N-terminal domain of apolipoprotein A-I as the most likely region for peptide interaction and suggested a strong connection between CIGB-814 and apolipoprotein A-I. Further validation by molecular dynamics simulations suggests that CIGB-814 may enhance the flexibility of structural areas in apolipoprotein A-I critical for high-density lipoproteins assembly. Additionally, an affinity enzyme-linked immunosorbent assay (ELISA) confirmed that apolipoprotein A-I has a tenfold greater affinity for CIGB-814 compared to the wild-type peptide. These integrated experimental and computational findings demonstrate that a single residue substitution is critical for apolipoprotein A-I recognition. In addition, this study provides mechanistic insight into CIGB-814 immunomodulatory and metabolic effects, expanding the therapeutic potential of Jusvinza to metabolic diseases associated with lipid homeostasis imbalance.
This study assessed the comprehensive human risk exposure to metal(loid)s through consumption of fish and clams from the Nun River. Thirteen elements (Ag, As, Ba, Cd Co, Cr, Cu, Fe, Mn, Ni, Pb, V, and Zn) were determined in the aquatic samples using inductively coupled plasma-optical emission spectrometry (ICP-OES). Metal(loid) data were subjected to simple descriptive and inferential statistical analysis to examine variations among species and sampling periods. In fish, Fe was the dominant element, while Zn and Mn showed temporal increments. Toxic metal(loid)s such as Pb, Cr, Cd, Ni, and As were detected at concerning levels, with some concentrations exceeding the international safety thresholds of the joint Food and Agriculture/World Health Organization. Species differences were observed, with higher accumulation in carnivorous fish, while clams generally showed lower levels. Health risk assessment indicated that most hazard quotient (HQ) values were below 1.0, suggesting generally low noncarcinogenic risk under the exposure assumptions applied; however, elevated values were observed for Cd and As in some fish species. Cancer risk analysis identified Cd and As as the major contributors to lifetime carcinogenic risk. This study provides a comprehensive and integrated assessment of metal(loid) contamination in Nun River fish and clams, examining temporal variation, species-specific accumulation patterns, and associated human health risks. In doing so, it helps to address a significant regional data gap. The research findings highlight potential exposure concerns and emphasize the need for continued monitoring, pollution control, and public awareness to support food safety in the Niger Delta region.
Pillar-[6]-arene 1 2- is a dianionic and accessible host, possessing a cylindrical cavity composed of six benzene rings with ten ethoxy and two carboxylic acid groups at the top/bottom portals. The results from 1H NMR spectroscopy, dynamic light scattering (DLS), scanning electron microscopy (SEM), and transmission electron microscopy (TEM) measurements of bolaamphiphilic 1 2- in 30 mM phosphate buffer (PBS) at pH = 7.4 are in line with its self-association into spherical nanoparticles at concentrations up to 200 μM. At lower concentrations (<25 μM), nanoparticles of 1 2- are smaller (D = 150 nm, DLS), while at the 25-100 μM concentration range, they grow bigger (D = 330 nm, DLS). With computational studies (molecular mechanics) revealing the electronic and shape complementarity of the cavity of 1 2- and anticancer drugs mitoxantrone (MTO2+) and doxorubicin (DOX+), 1H NMR spectra of equimolar drug/pillararene mixtures resulted in a severe broadening of all signals. The formation of drug/pillararene noncovalent complexes was further confirmed by the pH-triggered precipitation of both drugs and pillararene from their mixtures. The results from UV-vis supramolecular titrations were in line with two equilibria characterizing noncovalent complexations and corresponding to the formation of (a) multimeric [drug x ⊂ y ] complexes (x > y) along with (b) binary and inclusion [drug⊂pillararene] complexes. The existence of the latter was corroborated with mass spectrometry (ESI/TOF MS). Importantly, multimeric [drug x ⊂pillararene y ] and binary [drug⊂pillararene] complexes constitute sizable and spherical nanoparticles (600 nm-1 μm; DLS and TEM). With the biocompatibility of nanoparticulate 1 2- toward HeLa cells and its capacity to capture toxic anticancer agents used in clinics thereby forming larger nanoparticles, we reason that the nanosystem described here could be developed into a platform for effective sequestration or delivery of anticancer drugs.
In recent years, advancements in metabolomics have highlighted that changes in metabolic states within the body might also be closely related to missed abortion. However, the specific impacts of metabolic abnormalities on missed abortion remain unclear. This study screened differential metabolites in serum and villous tissues from patients with missed abortion, analyzed possible mechanisms linking metabolic disturbances to disease occurrence, and explored their clinical research value in early pregnancy abnormality research. Focusing on the effect of abnormal levels of metabolites on missed abortion, a prospective case-control study was conducted including 78 women with missed abortion and 75 women with normal early pregnancy undergoing elective termination. Targeted metabolomics profiling of serum and villous tissues was performed using mass spectrometry. Clinical characteristics and metabolite levels were compared between groups. Firth's logistic regression was applied to assess independent associations. ROC curve analysis was used to evaluate the discriminatory potential of key metabolites. Results revealed metabolic differences in both serum and villous tissues between the missed abortion and control groups. Serum was characterized by upregulated amino acid metabolism, with elevated levels of multiple amino acids including ORN and TYR, which were associated with the risk of missed abortion. Villous tissues exhibited core disturbances in acylcarnitine metabolism, with C10:1 and C18 significantly upregulated in the missed abortion group. Notably, no highly consistent differential metabolites were identified between serum and villous tissues. This study demonstrates distinct metabolic signatures in missed abortion, with amino acid upregulation in serum and acylcarnitine disorder in villous tissues. Only CIT was consistently upregulated in both matrices, possibly due to limited assay coverage. Combined panels of the top eight FDR-ranked metabolites achieved high AUCs in serum and villous tissue, offering good auxiliary discrimination for missed abortion. These findings provide new insights into its metabolic pathogenesis.
Dissolved organic matter (DOM) plays a central role in soil carbon (C) cycling as the most mobile and reactive C fraction in forests, regulating the microbial metabolism, nutrient availability, and C export. However, molecular-level DOM responses to environmental stressors such as warming and nitrogen (N) deposition remain poorly constrained, particularly under their combined influences. Thus, we investigated how 14 years of soil warming, N-addition, and combined heat + N influence soil-derived DOM quantity and chemistry. Using solution-state NMR spectroscopy and Fourier transform ion cyclotron resonance mass spectrometry, we resolved DOM composition across molecular size, biochemical class, mobility, and oxidation state. While the DOM quantity remained unchanged, warming enhanced microbial processing and oxidative transformation, yielding DOM enriched in oxidized, structurally complex compounds, whereas N-addition suppressed decomposition, limiting the release of plant-derived biopolymers and shifting DOM toward more microbial-derived constituents. Heat + N produced the most compositionally diverse DOM, with molecular shifts more closely resembling warming-induced responses, indicating that temperature-driven decomposition dominates under interacting stressors. These results demonstrate that chronic warming and N addition influence C cycling through distinct, yet nonadditive molecular pathways not captured by single-factor studies. This underscores the necessity of multifactor experiments to accurately capture the current and future ecosystem responses to interacting environmental stressors.
In previous studies, we identified the unique tumor immunosuppressive function of IgG4. This study aims to further characterize the structure of IgG4 and evaluate its potential effects on monocyte-derived dendritic cells (moDCs). The densities of IgG4 and CD11c in esophageal cancer tissues were detected by immunohistochemistry. IgG1 and IgG4 subclasses were purified from IgG1 and IgG4 subclasses were purified from intravenous immunoglobulin (IVIg) and human serum samples using subclass‑specific affinity chromatography. Human peripheral CD14⁺ monocytes were isolated and induced to differentiate into moDCs in vitro. The migratory and phagocytic capacities of moDCs were then evaluated following stimulation with IgG1 and IgG4. Immunohistochemical staining revealed that the densities of IgG4⁺ and CD11c⁺ cells were significantly higher in esophageal squamous cell carcinoma (ESCC) tissues than in adjacent normal tissues. Protein electrophoresis, molecular weight analysis, lectin affinity chromatography, and spectrometry demonstrated that native IgG4 exhibits distinct glycosylation profiles compared with IgG1 or commercial myeloma-derived IgG4. IgG4 was predominantly enriched in the ConA⁺ IgG fraction, indicating that it undergoes extensive high-mannose glycosylation. Immunocytochemical staining revealed that peripheral blood-derived monocytes expressed CD68 and CD206, and displayed strong binding affinity for IgG1 but weak affinity for IgG4. In vitro co-culture of moDCs with IgG4 revealed that IgG4 significantly promoted the migration and phagocytic capacity of moDCs relative to IgG1. Furthermore, IgG4 purified from both IVIg and patient serum exerted comparable effects on moDC differentiation. Collectively, these findings suggest that IgG4-associated glycosylation features may be linked to altered phenotypic and functional parameters of moDCs in this exploratory in vitro model. However, the underlying mechanisms, tissue-level relevance, and biological significance require further validation in larger cohorts and more comprehensive functional studies.
The accurate and rapid determination of trace copper Cu-(II) ions in environmental water is of great significance for both ecological safety and human health. In this work, a novel, eco-friendly, and highly sensitive fluorescence sensor was developed using carbon dots synthesized from pine cone deep eutectic solvent (DES) lignin and doped with lysine for the selective detection of Cu-(II) ions. The synthesized carbon dots exhibited excellent optical properties. They demonstrated a strong, concentration-dependent fluorescence response to Cu-(II) with a wide linear range of 2.81 × 10-7-5.87 × 10-5 and a remarkably low limit of detection (LOD) of 2.8 × 10-8 mol L-1. To validate the practical applicability and reliability of the proposed sensor, it was successfully employed for Cu-(II) determination in highly complex matrices, including various certified reference materials (primary drinking water, NASS-5 seawater, SM-WW1, and SM-WW2 wastewater) and real environmental samples (ground, lake, and tap water). The standard addition method yielded excellent recovery rates ranging from 98.9 to 104.4%, with relative standard deviations (RSDs) strictly below 4.3%. Furthermore, the analytical results were statistically compared with the standard flame atomic absorption spectrometry (FAAS) method at a 95% confidence level, showing exceptional agreement and no significant differences. The proposed lysine-doped carbon dot sensor offers a highly accurate, cost-effective, and highly selective alternative for the routine monitoring of Cu-(II) in diverse and complex environmental water samples, effectively overcoming significant matrix interference.
This paper investigates the concentrations of heavy metals in apricots, which are widely consumed as food and may pose serious health risks when exceeding permissible limits. The elements analyzed included lead (Pb), copper (Cu), zinc (Zn), arsenic (As), selenium (Se), cadmium (Cd), and mercury (Hg). The effects of microwave optimization digestion (MWOD) parameters, including temperature (tp), time (tm), and power (pw), on the extraction efficiency of these elements were systematically evaluated in apricot kernel (K), kernel shell (S), and flesh (F) samples. Microwave digestion was performed according to EPA Methods 3051A and 3052. Following digestion and complete solubilization, the concentrations of heavy metals were determined using inductively coupled plasma mass spectrometry (ICP-MS). The highest concentrations of Cd and Hg in apricot flesh were found to be 25.49 μg kg-1 and 14.21 μg kg-1 under optimized digestion conditions of 1000 W at 220 °C for 20 min and 800 W at 220 °C for 20 min, respectively. Pearson correlation analysis revealed a strong positive relationship between Zn and Cu under the applied digestion conditions. The accuracy and reliability of the analytical results were validated using the certified reference material BCR-414 (plankton). In addition, the potential health risks associated with the detected heavy metals were assessed using Estimated Daily Intake (EDI) and Hazard Quotient (HQ) models. The optimization of microwave digestion parameters significantly influenced extraction efficiency, and the combined use of optimized digestion and ICP-MS analysis enabled reliable identification and quantification of toxic elements in different apricot partitions.
Hepatocellular carcinoma (HCC) cells are metabolically reprogrammed for excessive uptake and metabolism of many nutrients. The tumor suppressive microRNA-148a-3p (miR-148a-3p) is downregulated in HCC, whereas its function in regulating HCC cell metabolism remains obscure. Herein we aimed to delineate the role of miR-148a-3p in HCC cell metabolism by using novel bioengineered miR-148a-3p (BioRNALeu/miR-148a-3p) agent produced in vivo. BioRNALeu/miR-148a-3p was designed by using human leucyl transfer RNA fused hsa-pre-miR-34a carrier, overexpressed in Escherichia coli (E. coli), and purified to high homogeneity. After transfection into HCC cells, the released miR-148a-3p levels were assessed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Cell proliferation was determined by CellTiter-Glo assays. Targets were validated by dual-luciferase reporter assays, immunoblotting, and immunofluorescence confocal imaging. Glycolysis capacity was evaluated by Seahorse XF assays, and glucose, lactate, and amino acid levels were quantified by liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods. BioRNALeu/miR-148a-3p was efficiently processed into target miR-148a-3p in HCC cells to effectively inhibit cell proliferation in a dose- and time-dependent manner. Mechanistically, miR-148a-3p suppressed the protein levels of glucose transporter GLUT1/SLC2A1 and L-type amino acid transporter LAT1/SLC7A5 via acting on their 3'-untranslated regions, as well as amino acid transporter ASCT2/SLC1A5. These, in turn, led to a reduction of glucose uptake, lactate production, and glycolytic flux in HCC cells, and alteration of intracellular amino acid metabolome including glutamine, leucine, phenylalanine, tyrosine, and methionine. Reintroduction of miR-148a-3p into HCC cells modulates glucose and amino acid metabolism via regulating multiple SLC transporters, thereby suppressing HCC cell viability. These findings highlight the role of miR-148a-3p in HCC cell metabolism and potential of bioengineered miRNA molecules for functional studies and therapeutic development.
Somatic embryogenesis triggers epigenetic changes that influence catechin accumulation in lingonberry, helping to improve phytochemical quality in micropropagated plants. Vaccinium vitis-idaea L. (lingonberry), a boreal superfruit valued for its high antioxidant content, exhibits high potential for regeneration via somatic embryogenesis (SE). However, in-vitro culture often induces epigenetic variations that can influence phytochemical stability. To find out whether DNA methylation plays a role during SE, we compared DNA methylation levels in the regenerants of two lingonberry genotypes, Erntedank (Cv1) and a hybrid designated as "H1" (identity undisclosed). We demonstrate that SE in lingonberries induces epigenetic modifications, particularly epigenetic memory and biological imprinting that contribute to enhanced catechin accumulation, as revealed by highly sensitive DNA methylation profiling using ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS/MS) and catechin analysis via proton nuclear magnetic resonance (1H NMR) across four experimental groups: in-vitro-derived shoots, in-vitro and ex-vitro SE-regenerated plants, and controls. The optimized UHPLC protocol enabled robust quantification of 5-methylcytosine (5mC), revealing significantly elevated DNA methylation in in-vitro-derived shoots (Cv1-TC: 84.02%, H1-TC: 87.30%) compared to ex-vitro SE-regenerated plants (Cv1-GC: 53.79%, H1-GC: 60.27%). Parallel 1H NMR analysis confirmed the presence of catechins, with ex-vitro SE plants showing the strongest signals across aromatic (6.93-6.06 ppm), sugar (4.57-4.01 ppm), and aliphatic (2.93-2.54 ppm) regions. In-vitro-derived shoots exhibited high methylation but minimal catechin signals, suggesting epigenetic suppression of flavonoid biosynthesis under culture-induced stress. This negative relation indicates that the methylation status influences secondary metabolite profiles. The findings underscore the importance of monitoring epigenetic and biochemical integrity during plant regeneration. This dual-platform strategy provides a valuable tool for optimizing micropropagation systems, monitoring epigenetic changes, and improving phytochemical consistency in lingonberry.
A green and sustainable strategy was employed for the in situ formation of silver nanoparticles (AgNPs) within a TEMPO-oxidized cellulose nanofibril (CNF) matrix, using CNF as both a reducing and stabilizing system. AgNPs formation was confirmed by the presence of a surface plasmon resonance band at approximately 420 nm, while DLS indicated colloidal dispersion and transmission electron microscopy (TEM) revealed predominantly spherical nanoparticles, mostly in the 10-20 nm range, associated with the CNF network. The CNF-AgNP nanocomposite dispersion was further processed into hydrogels, films, and aerogels, which were characterized by transmission electron microscopy (TEM), rheology, tensile testing, field-emission scanning electron microscopy (FE-SEM), and thermogravimetric analysis (TGA). Further, the actual silver content in the CNF-AgNP hydrogel was determined by Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES). The hydrogel exhibited shear-thinning behavior, whereas the films showed a tensile strength of 1.39 MPa and thermal stability up to approximately 100 °C. The CNF-AgNP composites inhibited the growth of Pseudomonas aeruginosa, Staphylococcus aureus, and Bacillus subtilis through disk diffusion assays, while CNF alone showed negligible antibacterial activity. In the Zebrafish (Danio rerio) Embryo Toxicity Test (FET Test), end points such as embryo coagulation, lack of somite formation, nondetachment of the tail, and absence of heartbeat were assessed for CNF-AgNPs that induced acute lethality (LC50) of 1.115 μM at 96 h, whereas CNF dilutions were nontoxic under the tested conditions. Overall, TEMPO-oxidized CNF enabled the fabrication of multifunctional CNF-AgNP hydrogels, films, and aerogels with antibacterial activity and reduced acute embryotoxicity compared with free silver ions. Further studies addressing silver release kinetics, long-term exposure, and performance under real-world conditions are warranted to substantiate their environmental and biomedical safety.
The reaction of the ten-vertex hydridorhodathiaborane [2,2,2-(H)(PPh3)2-closo-2,1-RhSB8H8] (1) with excess of carbon monoxide at room temperature yields a novel eleven-vertex rhodathiacarborane [1,1-(CO)(PPh3)-3-(OH)-closo-1,2,3-RhSCB8H8] (4). The formation of 4 involves the substitution of a PPh3 ligand by CO at the rhodium centre and the subsequent insertion of a second CO molecule into the polyhedral cage to form a hydroxy-functionalized carbon vertex (C-OH). In addition to 4, the reaction affords the CO-ligated ten-vertex adducts [6,6-(CO)(PPh3)-9-(PPh3)-arachno-6,5-RhSB8H9] (5a) and [9,9-(CO)(PPh3)-8-(PPh3)-arachno-9,6-RhSB8H9] (5b). While a mixture of 5a and 5b can also be synthesized from the PPh3-adduct [6,6-(PPh3)2-9-(PPh3)-arachno-6,5-RhSB8H9] (2), compound 2 does not yield 4 upon treatment with CO. This suggests that the transformation of CO into a C-OH vertex is mediated by a specific CO-Rh/PPh3 intermediate. Similarly, the reaction of the 2-methyl-pyridine adduct, [6,6-(PPh3)2-9-(2-Mepy)-arachno-6,5-RhSB8H9] (3) with CO selectively yields the CO-ligated isomer [6,6-(CO)(PPh3)-9-(2-Mepy)-arachno-6,5-RhSB8H9] (6). All the compounds were characterized by multielement NMR spectroscopy and mass spectrometry, with 4, 5a and 6 further analysed by single-crystal X-ray diffraction. Variable temperature NMR and DFT calculations for 4 reveal a fluxional process characterized by the rotation of the {Rh(CO)(PPh3)} unit relative to the thiacarborane cage. These findings underscore the versatility of rhodathiaborane platforms for the activation and incorporation of small molecules via metal-polyhedron cooperation.
Peritoneal dialysis (PD) is a cornerstone kidney replacement therapy for patients with end-stage kidney disease; however, chronic exposure to bioincompatible dialysis solutions progressively damages the peritoneal membrane, leading to mesothelial-to-mesenchymal transition, fibrosis, and ultimately ultrafiltration failure. Currently, peritoneal membrane dysfunction is detected only at advanced stages through the peritoneal equilibration test, underscoring the need for earlier and more sensitive biomarkers. Extracellular vesicles (EVs) isolated from peritoneal dialysis effluent (PDE) have emerged as promising candidates, given their capacity to carry proteins, lipids, and nucleic acids that reflect local and systemic cellular activity. To date, 12 studies have successfully isolated and characterized PDE-derived EVs, identifying canonical EV markers, such as CD9, CD63, CD81, TSG101, and HSP70, alongside disease-relevant molecules, including galectin-3 binding protein, aquaporin-1, glycoprotein 96, and integrin-linked kinase. These EVs are enriched in signaling components associated with inflammation, angiogenesis, and fibrosis, particularly through TGF-β/p38 and NF-κB pathways. Moreover, specific EV-associated microRNAs (e.g., miR-125a-5p, miR-132-3p, miR-296-3p, miR-432-5p) overlap with molecular signatures observed in kidney and cardiometabolic disorders, suggesting broader systemic relevance. As an original contribution to this review, and to address cross-study comparability, we applied EVqualityMS, a mass spectrometry-based quality assessment tool, to benchmark EV enrichment and contaminant profiles and calculate quality indices across publicly available PDE-EV proteomics datasets. Despite methodological heterogeneity and limited sample sizes, PDE-derived EVs represent a powerful "liquid biopsy" of the peritoneal environment. Their integration into PD monitoring holds promise for the early detection of membrane injury and for supporting a predictive, biomarker-guided, and personalized approach to PD management.
While fucoidans show anti-atherosclerotic potential, their impact on this gut-host axis is unclear. This study investigated the effects of Fucus vesiculosus fucoidan, alone and combined with simvastatin, on gut microbiota and metabolome in an atherosclerotic rabbit model that closely recapitulates key features of human atherosclerotic pathogenesis. Atherosclerosis was induced in New Zealand rabbits via high-fat diet feeding combined with balloon catheter injury. Animals in the experimental groups (n = 6 per group) were treated with either F. vesiculosus fucoidan alone (100 mg/kg) or a combination of fucoidan (100 mg/kg) and simvastatin (5 mg/kg). Lipid profiles were assayed using commercial kits, and aortic pathological changes were evaluated by Oil Red O staining. Genomic DNA from intestinal contents was analyzed by polymerase chain reaction, and untargeted metabolomics were performed using liquid chromatography-tandem mass spectrometry. Treatments significantly reduced atherosclerotic plaque formation. This effect was particularly pronounced when fucoidan was combined with simvastatin, reducing plaque formation from 41.77 ± 16.02% to 5.91 ± 8.03% in the abdominal aorta and from 10.72 ± 3.49% to 2.29 ± 2.30% in the thoracic aorta (Model vs. combination group). The treatment also ameliorated hyperlipidemia, as shown by decreased plasma TC (24.55 ± 0.73 to 17.45 ± 0.58 mmol/L) and TG (7.75 ± 0.46 to 0.83 ± 0.25 mmol/L) in the combination group compared to the Model group. Both intervention groups exhibited enhanced microbial diversity and increased species richness across all taxonomic levels compared to the model group. Moreover, fucoidan and combination treatments significantly upregulated 83 and 128 metabolites and downregulated 125 and 121 metabolites, respectively. KEGG enrichment analysis indicated that these metabolic changes were associated with multiple pathways. Moreover, the combination therapy may mitigate certain side effects associated with simvastatin, although this possibility requires further investigation. The combination of F. vesiculosus fucoidan and simvastatin holds promising therapeutic potential for preventing and treating atherosclerosis. Our findings provide novel evidence that this enhanced effect is likely linked to structural and functional modifications in the gut microbiota and its associated metabolic profile, supporting a multifaceted strategy that extends beyond lipid-lowering.