Search Results (28,114)

Wuweiganlu (WGL) is a traditional formulation widely applied in the treatment of rheumatoid arthritis (RA), yet the identity of its bioactive constituents remains inadequately defined. In this study, ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) and untargeted serum metabolomics were employed to characterize the active components of WGL. Fraxin was identified as a principal compound from WGL. To investigate its therapeutic mechanism in RA, a series of in silico and experimental approaches were conducted. Network pharmacology analysis and RNA sequencing identified heat shock protein family member 8 (HSPA8) as a potential molecular target of Fraxin, which was further validated by molecular docking studies. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses indicated that Fraxin exerts its effects primarily by modulating cell apoptosis through the PI3K signaling pathway. In vitro experiments demonstrated that Fraxin significantly reduced inflammatory responses and downregulated HSPA8 expression in lipopolysaccharide (LPS)-stimulated fibroblast-like synoviocytes (FLs) and macrophages. In vivo, Fraxin administration markedly reduced paw swelling, alleviated bone deformities, and improved bone volume fraction (BV/TV) in male IL1RA-deficient mice exhibiting spontaneous arthritis. Histological analysis confirmed that Fraxin attenuated joint inflammation by modulating the inflammatory microenvironment. Additionally, Fraxin inhibited synovial hyperplasia by regulating mitochondrial membrane potential collapse in FLs. Functional assays revealed that this regulation occurred via the inhibition of HSPA8/PI3K/AKT signaling axis, thereby suppressing aberrant FLS proliferation and contributing to the attenuation of RA progression. Full article

Oral secretions (OS) of the Asian corn borer (Ostrinia furnacalis) play pivotal roles in its interactions with host plants. To systematically characterize the composition and potential functions of OS, 245 functionally annotated proteins from O. furnacalis reared on artificial diet were identified using an integrated approach combining transcriptomics and liquid chromatography–tandem mass spectrometry (LC-MS/MS). Bioinformatics analyses were further performed to predict 16 effector proteins. Subsequent tissue-specific expression assays were conducted to quantify the transcriptional levels of genes encoding these effector proteins in salivary glands, guts, and residual tissues. Among these genes, OfGDH2, OfPero2, OfPero3, and OfCBP were highly expressed in salivary glands, OfGDH3, OfRGD, OfEST1, OfEST2, OfMET, and Offerrin showed high expression in guts, while OfSP34 was abundantly expressed in both salivary glands and guts. This study represents the first systematic characterization of the molecular profile of O. furnacalis OS, thereby laying a solid foundation for future investigations into the molecular mechanisms underlying the host adaptation of this pest. Full article

The effects of static magnetic field (SMF) treatment on the solid-state culture of Sanghuangporus vaninii (SV) were investigated to enhance metabolite production and bioactivity. SMF parameters including intensity, exposure duration, and temperature were optimized, and treatment at 4 mT for 2 h per day produced the most pronounced effects, increasing total flavonoid (TFC), polyphenol (TPC), and triterpenoid (TTC) contents by 61–438% compared with the control. Ultrasonic extraction and semi-preparative chromatography enabled the isolation of three key compounds: D-(+)-trehalose (1), 5,7-dihydroxy-3,4′-dimethoxyflavone (2), and pinolenic acid (3), all of which were elevated following SMF treatment. Importantly, SMF exposure was associated with enhanced inhibitory activities against enzymes relevant to chronic metabolic disorders. The overall inhibitory activities against α-amylase, α-glucosidase, pancreatic lipase, and xanthine oxidase increased by 6–28% compared with the control, reaching a maximum inhibition of 97.60 ± 0.17%. Preliminary in vitro screening at 100 μg/mL showed that compounds 1 and 2 inhibited both α-amylase and α-glucosidase, whereas compound 3 selectively inhibited pancreatic lipase. Subsequent IC₅₀ analysis confirmed that compound 2 under SMF treatment exhibited inhibitory activity comparable to acarbose against α-amylase (45.62 μg/mL vs. 52.18 μg/mL) and α-glucosidase (38.74 μg/mL vs. 35.42 μg/mL). In addition, compound 3 showed moderate inhibition of pancreatic lipase with an IC₅₀ value of 42.15 μg/mL. These findings suggest that SMF treatment may enhance metabolite production and in vitro enzyme inhibitory activity in S. vaninii. However, these results are limited to in vitro assays, and further studies including cellular and in vivo validation, toxicity assessment, and pharmacokinetic evaluation, are required before any therapeutic or industrial applications can be considered. Full article

The Asian citrus psyllid Diaphorina citri Kuwayama (Hemiptera: Liviidae) is the vector of the devastating citrus disease Huanglongbing, posing a significant threat to the global citrus industry and necessitating environmentally sound management strategies. This study aimed to evaluate Australian tea tree oil (TTO) and its primary constituents as potential botanical insecticides. Gas chromatography-mass spectrometry (GC-MS) was performed to analyze the chemical profile of commercial TTO, and behavioral effects on D. citri adults were assessed using a Y-tube olfactometer. Direct spray bioassays were conducted to determine contact toxicity. A total of 12 compounds were identified, with TTO being a Terpinen-4-ol chemotype, dominated by Terpinen-4-ol (40.62%), γ-Terpinene (21.46%), and α-Terpinene (10.45%). TTO demonstrated potent, concentration-dependent repellency, achieving 100% repellency at 10 g/L. In contrast, Terpinen-4-ol alone was attractive to psyllids at low concentrations, suggesting synergistic or masking effects within the complex oil blend. TTO and its major constituents also exhibited significant dose- and time-dependent contact toxicity. Although the 72 h LC₅₀ of TTO (19.18 g/L) indicates lower potency compared to conventional insecticides (0.59–1.23 g/L), its combined repellent and toxic properties make it a promising candidate for integrated pest management (IPM) programs aimed at controlling D. citri and mitigating insecticide resistance. Full article

Background/Objectives: Wound healing is a complex biological process involving inflammatory, proliferative, and remodeling phases. Plant-derived essential oils are increasingly investigated as topical therapeutic agents, although their biological effects are strongly influenced by composition and formulation. The present study evaluated the effects of topical Mentha spicata essential oil on cutaneous wound healing in a rat excisional wound model and explored potential molecular mechanisms using a network-based bioinformatic approach. Methods: Twenty-one male Wistar rats were randomly assigned to three groups and treated twice daily for 14 days with a formulation containing 5% Mentha spicata essential oil diluted in olive oil, olive oil alone, or no treatment. Wound healing was assessed through macroscopic monitoring and histological scoring. The chemical composition of the essential oil was characterized using gas chromatography–mass spectrometry analysis. Predicted molecular targets of the major monoterpenes were analyzed through protein interaction networks and pathway enrichment analysis. Results: Macroscopic wound closure progressed in all groups by day 14. Histological analysis revealed that the olive oil group showed more advanced collagen deposition, re-epithelialization, and granulation tissue maturation, whereas the Mentha spicata group displayed a more pronounced inflammatory and proliferative histological pattern. Network-based analysis highlighted signaling pathways related to receptor-mediated cellular responses as potential molecular mechanisms associated with early inflammatory and proliferative processes. Conclusions: These findings suggest that the biological effects of Mentha spicata essential oil in wound repair may be phase-dependent and influenced by concentration and formulation. The results support further studies aimed at optimizing dose and delivery strategies for essential oil–based wound therapies. Full article

The poly(A) tail of mRNA plays a vital role in mRNA transcript stability, translational efficiency, and immunogenicity. Co-transcriptionally polyadenylated in vitro transcribed (IVT) mRNAs typically contain poly(A) tails of 50–120 nucleotide tail length due to limitations in production of template pDNA with longer poly(A) sequences. In contrast, post-transcriptional enzymatic polyadenylation of mRNA with poly(A) polymerase (PAP) presents a modular alternative to increase the tail length. However, the lack of real-time control strategies for PAP-mediated tailing has limited its broader applicability in mRNA production. Here, we develop a methodology for controlling poly(A) tail length in post-transcriptional polyadenylation of mRNA that uses adenosine triphosphate (ATP) consumption measured at-line to predict the poly(A) tail length. We establish a novel analytical method based on monolith reverse-phase chromatography to validate the poly(A) predictions. We were able to produce longer poly(A) tails and accurately determine their length in 300–700 nt range. The resulting longer poly(A) tailed reporter mRNAs outperformed the encoded and shorter poly(A) tailed mRNAs in cell-based assays. This work presents a new strategy for controlled post-transcriptional polyadenylation using ATP consumption as a process control metric, an approach which may in future be expanded to other NTP-dependent enzymatic conversions. Full article

This study investigated the antibiofilm and anti-quorum-sensing (QS) potential of endophyte extracts isolated from medicinal plants and their validation against Pseudomonas aeruginosa. Endophytes were isolated from the plants using the serial dilution method, and the extracts produced by these endophytes were screened for antimicrobial and biofilm-inhibition activity using assays. The efficient extract was biochemically characterized, followed by validation of its secondary metabolite content. Furthermore, QS-regulatory gene expression levels and microscopy were used to confirm inhibition of biofilm formation. A total of 12 cultures, including 8 bacterial and 4 fungal, were isolated and screened, demonstrating efficient antimicrobial activity (zone of inhibition of 18.8 mm) and 64.3% antibiofilm activity. The efficient endophyte isolated from Paederia foetida was identified as Bacillus xiamenensis MM07 by 16S rRNA gene sequencing. MM07 extract analyses by biochemical and Fourier transform infrared methods revealed the presence of diverse biomolecules. A dose-dependent inhibition was observed, achieving up to 83.5, 60.3, 73.2, 82.7, 83.2, and 15.1 in biofilm formation and exopolysaccharide, violacein, pyocyanin, protease, and alginate production, along with 63.2% swimming ability at 30 µg/mL, respectively. Gas chromatography-mass spectrometry analyses validated the presence of major secondary metabolites, including 3,3-dimethyl-4-methylamino-butan-2-one, 6-amino-2-methyl-, 1-iodo-2-methylundecane, and hexadecanoic acid, with the potential to inhibit biofilm and QS activity. Quantitative real-time polymerase chain reaction of QS regulatory genes (lasI, lasR, rhlI, and rhlR) and microscopy analysis confirmed the anti-QS properties, evidenced by a 40.3% decline in gene expression and biofilm inhibition by MM07 extract. These findings highlight the potential of novel B. xiamenensis MM07 endophyte from P. foetida as a sustainable source of biomolecules for combating biofilm-associated infections. Full article

Caffeine is widely consumed and generally considered safe at customary doses. How-ever, high-dose preparations available online pose a risk of severe and potentially fatal intoxication. Although uncommon, lethal caffeine poisoning is associated with profound cardiovascular and neurological toxicity. A rare case of intentional acute caffeine intoxication with fatal outcome is presented. A 25-year-old woman ingested an estimated 60 tablets containing 200 mg of caffeine each, purchased online. She was admitted to hospital shortly after ingestion of the caffeine tablets with palpitations, agitation, dizziness, and repeated vomiting. On examination, she presented with arterial hypotension (90/60 mmHg) and marked sinus tachycardia (150 beats/min), accompanied by psychomotor agitation. Her blood caffeine concentration measured by means of high-performance liquid chromatography (HPLC) was 177 µg/mL. The patient’s condition rapidly deteriorated, with the development of convulsive syndrome progressing to coma, extreme ventricular tachycardia, exotoxic shock, and toxic cardiomyopathy. Despite intensive care management, including mechanical ventilation and advanced cardiopulmonary resuscitation, the patient died several hours after admission. In conclusion, this case underscores the life-threatening potential of acute high-dose caffeine ingestion and highlights the risk associated with unrestricted access to concentrated caffeine products. Early recognition and aggressive management are crucial, yet may be insufficient in cases of massive overdose. Full article

In this study, the active components in the seed of Mangifera indica L. were isolated, the main chemical components were identified, and then their antioxidant activities and their effects on liver injury and intestinal microbiota were evaluated. The results showed that all the components of mango column chromatography exhibited antioxidant activity. F2 had the lowest IC₅₀ value of 93.61 μg/mL and exhibited the strongest DPPH radical scavenging activity. Given its superior overall antioxidant activity, F2 was selected for further compositional analysis and activity evaluation. UPLC-MS/MS analysis showed that the isolated components of mango F2 contained 11 active ingredients, including mangiferin, gallic acid and quercetin. The results showed that specific mango fractions significantly reduced serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, and showed a protective effect on liver injury induced by alcohol. rRNA sequencing analysis showed that high alcohol intake could reduce the species diversity of intestinal microbiota in mice, and mango fractions could effectively alleviate this phenomenon. High alcohol intake decreases the relative abundance of Bacteroidota and increases the abundance of Bacillota and Thermodesulfobacteriota phyla. The high-dose mango group alleviated the above changes, which was manifested by an increase in the relative abundance of Bacteroidota and Thermodesulfobacteriota bacteria. The relative abundance of families such as Muribaculaceae in the high-dose mango group decreased compared to the model group. This study provides a scientific basis for the analysis and high-value utilization of mango components, and provides a new alternative for protecting against alcoholic liver injury and regulating intestinal microbiota. Full article

Agapanthus africanus (L.) Hoffmanns. is a medicinal plant traditionally used in South Africa for its promise as a source of bioactive compounds with anticancer properties. This study aimed to investigate the apoptotic effects of A. africanus fractions on cancer cell lines and to identify the bioactive phytochemical constituents using gas chromatography-mass spectrometry analysis. To test for cytotoxicity, MCF-7, A549, and HeLa cancer cells were treated with crude extract, n-hexane, n-butanol, dichloromethane, and aqueous fractions of A. africanus extracts at different concentrations (0.00–1000 µg/mL). Total apoptosis was quantified using Annexin V/PI staining. The 4′,6-diamidino-2-phenylindole was used to detect nuclear morphological changes and the Caspase-GLO 3/7 assay was employed to check the caspase activation in the cancer cells. Expression of apoptosis-related (caspase-3, bax, bcl-2) genes was evaluated using real time-polymerase chain reaction. The crude extract of A. africanus exhibited dose-dependent cytotoxicity against MCF-7, A549, and HeLa cells, with IC₅₀ values of 130 µg/mL, 380 µg/mL, and <125 µg/mL, respectively. Among the tested fractions, the n-butanol fraction showed cytotoxicity towards MCF-7 cells with an IC₅₀ value of <870 µg/mL. In contrast, n-hexane, dichloromethane and the aqueous fractions exhibited higher IC₅₀ values against cancer cells. Flow cytometry analysis, which was applied to quantify total apoptosis, revealed that the crude extract of A. africanus induced apoptosis by (~60%) compared to the n-butanol fraction, which exhibited a moderate apoptotic effect (~27%). DAPI nuclear staining showed nuclear shrinkage and chromatin condensation in the MCF-7 cell line, whereas in Caspase-GLO 3/7, the crude extract and n-butanol fraction resulted in significant luminescence, indicating activation of caspase-3/7. Caspase-3/7 analysis showed A. africanus treatments produced varying levels of apoptotic activation. The crude extract increased caspase activity by 2.9-fold, while the n-butanol fraction induced a 1.7-fold rise compared with untreated cells. GC-MS chromatograms detected and identified 16 compounds in the fractionated n-butanol and 23 compounds from the crude extract of A. africanus. The major compounds identified from the n-butanol fraction included n-hexadecanoic acid; α-tocopherol and 9,12,15-octadecatrienoic acid, while the GC–MS profile of the crude extract was dominated by 6,10,14-trimethylpentadecan-2-one; 1,3,5-Triphenylcyclohexane and phytol. The study indicates the pro-apoptotic potential of A. africanus, particularly in its crude form, supporting its ethnopharmacological use and suggesting its relevance as a candidate for anticancer drug discovery. Full article

The growing demand for efficient and cost-effective industrial proteases has intensified bioprospecting efforts in diverse ecosystems as a strategy to identify microorganisms with enhanced enzymatic performance. This study aimed to isolate, identify, and evaluate aerobic protease-producing bacteria from animal-protein matrices and water sources collected across the three continental regions of Ecuador, and to assess their suitability for industrial enzyme production A total of 34 bacterial strains were isolated and taxonomically assigned to the orders Enterobacterales, Pseudomonadales, and Bacillales. Proteolytic activity was evaluated using azocasein and casein assays after cultivation in an optimized medium containing 1% soybean paste as an inducer at 37 °C and 120 rpm for 72 h. Enterobacter cloacae (BC, pork), Bacillus paramycoides (P2, snook), and Pseudomonas aeruginosa (CH1, chontacuro) were identified as the most active protease producers from the Andean (Sierra), coastal (Costa), and Amazon regions, respectively. Production kinetics revealed marked strain-dependent differences. BC and P2 reached maximum proteolytic activity on day 4 followed by a decline, whereas CH1 peaked on day 2 and maintained stable activity over time, indicating superior enzymatic stability. Partial purification by gel-filtration chromatography (Sephadex G-100) yielded fractions with enhanced proteolytic activity, while SDS-PAGE analysis confirmed successful enrichment of protease-containing fractions. Overall, the results demonstrate that ecological origin strongly influences protease production and stability, and identify Pseudomonas aeruginosa CH1 as a particularly promising candidate for industrial applications requiring robust and sustained proteolytic activity. Full article

Acidithiobacillus caldus is perpetually exposed to multiple extreme environmental stresses. CsrA, functioning as a post-transcriptional regulator of physiological metabolism, acts as a differential modulator, facilitating more economical and efficient adaptation to extreme environments. The csrA expression recombinant strain was constructed in A. caldus MTH-04 by conjugative transfer technology pJD215. Physiological characterization revealed enhanced acid tolerance, significantly elongated flagella, elevated extracellular secretion, and altered biofilm composition. Notably, intracellular concentrations of free glutamate and aspartate increased to 24.18 mg/L and 16.07 mg/L, respectively. The secondary structure of CsrA protein was determined in vitro through circular dichroism spectroscopy and size-exclusion chromatography. Electrophoretic Mobility Shift Assay (EMSA) successfully demonstrated in vitro binding activity of CsrA to the rpoS leader mRNA. CsrA suppresses rpoS mRNA translation by competing with ribosomes for binding sites, thereby negatively regulating rpoS expression. Critical binding sites were further validated through site-directed mutagenesis. Through EMSA, RT-qPCR and the translation reporter system, it was also found that CsrA has a dual regulatory function for nearby flagella- and motility-related gene clusters (flgC, 07035, motD, 15040), which also implies the global regulatory role of CsrA. In summary, a potential overall post-transcriptional regulatory mechanism based on CsrA and rpoS by extremophile A. caldus was proposed. Finally, the efficiency of bioleaching application by csrA overexpression strain was improved by 20.81%. Full article

Background: Glutaric aciduria type-1 (GA-1) is a genetic disorder caused by glutaryl-coenzyme A dehydrogenase deficiency, leading to the accumulation of glutaryl-CoA and its derivatives. Clinical manifestations include neurological abnormalities; however, the underlying pathological mechanisms remain unclear. Early diagnosis and intervention are crucial for minimizing adverse outcomes. To date, diagnostic methods have certain limitations, and there is a critical need for a sensitive biomarker for diagnosis. We aimed to characterize metabolic dysregulation and identify candidate biomarkers associated with GA-1 in biochemically confirmed patients compared to age- and sex-matched control subjects. Methodology: Untargeted metabolomics profiling of GA-1 patients (n = 29) was compared to matched control subjects by age and sex. Multivariate and univariate statistical analyses were performed to identify dysregulated metabolites. Results: Our findings revealed 220 endogenous human metabolites. Notably, there was a strong enrichment in carboxylic acids and derivatives, including amino acids and derivatives, hydroxy and keto acids, fatty acyls, sphingolipids, phosphatidylcholines, and nucleotides and nucleosides. Pathway analysis indicates alterations in the biosynthesis of cardiolipin and phosphatidylcholine, as well as in pyrimidine metabolism, the urea cycle, and amino sugar metabolism. We demonstrated a robust performance model for 6-Methylnonanoyl-CoA, displaying strong discriminative power. Conclusions: We identified broad dysregulation across various biochemical classes, reflecting an imbalance in energy metabolism that involves carbohydrate and lipid pathways. The results also highlight dysregulation in sphingolipids, phospholipids, and nucleotide metabolism. These findings are preliminary and the clinical relevance of these findings in patients with GA-1 requires further investigation. We identified candidate biomarkers capable of distinguishing GA-1 patients from controls; however, these findings require validation in independent cohorts. Full article

Background: Protein A resins are indispensable for monoclonal antibody (mAb) production, yet their condition and performance are traditionally assessed using indirect or qualitative methods. In this study, the multi-attribute method (MAM), previously applied to therapeutic protein characterization, is systematically adapted for the first time as a unified liquid chromatography–mass spectrometry (LC-MS) platform for Protein A resin analysis. Method: Four Cytiva Protein A resins, MabSelect™, MabSelect SuRe™, MabSelect SuRe™ LX, and MabSelect™ PrismA, were evaluated by MAM for resin identity, Protein A ligand integrity, fouling by impurities, and cleaning performance. Results: MAM enables resin-specific peptide fingerprinting and quantitative monitoring of Protein A ligand post-translational modifications (PTMs), including deamidation, isomerization, and fragmentation induced by repeated clean-in-place (CIP) cycles. Comparative analysis of virgin and used resins revealed ligand degradation and fouling despite engineered alkaline stability, with MabSelect™ showing the greatest susceptibility. Importantly, residual monoclonal antibodies (mAbs) and host cell proteins (HCPs) were directly detected and quantified from the resin matrix, providing a molecular-level assessment of resin cleaning effectiveness not achievable with conventional approaches. Conclusions: This work establishes MAM as a novel, sensitive, and comprehensive strategy for Protein A resin lifecycle management, delivering actionable insight for resin selection, cleaning optimization, and downstream process development. Full article

This study investigated the bioactive properties of fractions derived from Rhynchosia nulubilis cultivated with Ganoderma lucidum mycelium (RNGM), focusing on cytotoxic and apoptosis-related responses in cancer cells. Fractions obtained using n-hexane, chloroform, ethyl acetate, and water were evaluated for cytotoxic effects against A549, Hep3B, HeLa, and HeLa229 cells using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The phenolic acid composition of each fraction was determined by high-performance liquid chromatography (HPLC). Among the fractions, the ethyl acetate fraction showed the highest total phenolic acid content and exhibited the strongest cytotoxic activity, particularly against HeLa cells. Apoptosis induction was supported by increased caspase-3/7 activity, apoptotic nuclear morphology observed by 4′,6-diamidino-2-phenylindole (DAPI) staining, and accumulation of cells in the sub-G1 phase. In addition, treatment with the ethyl acetate fraction upregulated p53 and Bax mRNA expression and increased the Bax/Bcl-2 ratio. These findings suggest that the ethyl acetate fraction of RNGM induces apoptosis-mediated growth inhibition in cervical cancer cells. Full article