Search Results (5,973)

Idelalisib (IDE) is a marketed chiral anticancer drug administered as the S-enantiomer, requiring sensitive monitoring of the R-enantiomer to ensure enantiomeric purity. However, no dedicated capillary electrophoresis (CE) method has been reported for trace-level quantification of R-IDE. In this study, a cyclodextrin-mediated CE method was developed for reliable detection of the R-enantiomer at the 0.1% level (LOD 2 µg/mL; LOQ 5 µg/mL). Systematic screening identified hydroxypropyl-β-cyclodextrin (HP-β-CD) with an intermediate degree of substitution (DS~6.8) as the optimal chiral selector, providing efficient enantioseparation (R_s up to 4.3). The method was validated according to ICH Q2(R2) guidelines, demonstrating suitable precision, accuracy, and robustness. Complementary NMR studies revealed hindered rotation of the 3-phenyl moiety and elucidated the molecular basis of enantioselectivity. Complexation with β-CD and HP-β-CD produced clear diastereomeric differentiation in both ¹H and ¹⁹F NMR spectra, while the simplified ¹⁹F NMR profiles enabled direct enantiomer discrimination. NOESY and ROESY experiments demonstrated distinct inclusion modes, with HP-β-CD accommodating both the fluorinated aromatic ring and the 3-phenyl moiety. These interactions may account for the superior enantioseparation observed with HP-β-CD of intermediate DS. Our validated CE method addresses the distomer determination while NMR insights provide mechanistic understanding of the chiral recognition. Full article

Cod liver oil is a rich source of polyunsaturated fatty acids (PUFAs) but is highly susceptible to oxidative degradation, limiting its use in topical formulations. This study aimed to develop stable cod liver oil nanoemulsions for topical application and to evaluated the influence of surfactant ratio (lecithin/PEG-15 hydroxystearate: 2.5:1 and 1:1, w/w), emulsification method (ultrasonication or high-pressure homogenization), and vitamin E acetate supplementation on their physicochemical properties and oxidative stability. Eight nanoemulsions were characterized in terms of droplet size, polydispersity, ζ-potential, vitamin E acetate encapsulation efficiency, oxidative stability, film-forming capacity and cytocompatibility. Among the investigated formulations, F4 (2.5:1 lecithin/PEG-15 hydroxystearate, high-pressure homogenization, with vitamin E acetate) exhibited the most favorable characteristics, including a mean droplet size of 67.95 nm, ζ-potential of −63.12 mV and vitamin E acetate encapsulation efficiency of 32.59%. The formulation demonstrated good physicochemical stability under thermal, mechanical and photostability testing, improved oxidative stability, transient film-forming behavior with an initial occlusive effect, and no cytotoxicity toward human dermal fibroblasts. These findings indicate that nanoemulsion performance depends on the combined influence of formulation composition and processing conditions, with F4 representing a promising topical carrier for cod liver oil intended for interaction with the stratum corneum. Full article

Background/Objectives: Pharmacists play a key role in managing urinary tract infection (UTI) symptoms by providing medications, self-care advice, and over-the-counter treatments, while referring patients to a doctor when necessary. This study aimed to examine the practices of community pharmacists in managing UTI symptoms and to gain insight into their attitudes toward antibiotic use for this condition. Methods: A cross-sectional study was conducted among community pharmacists in Serbia using a previously validated online questionnaire, assessed for content and face validity and pilot-tested among pharmacists. Results: A total of 430 community pharmacists participated in the study. Patients more often consulted pharmacists before visiting a doctor than after (median 5 vs. 3 per week; p < 0.001). For uncomplicated UTIs, pharmacists primarily recommended increased fluid intake (92.8%), herbal teas (94.7%), and food supplements (85.6%), whereas for complicated UTIs, most referred patients to a doctor (95.4%). Attitudes, perceived competence, and support for over-the-counter antibiotic availability were significantly associated with gender, years of experience, and specialization. Pharmacists who agreed that antibiotics are the most effective treatment for uncomplicated urinary tract infections were more likely to refer patients to a doctor (p < 0.01). Conclusions: Pharmacists are frequently consulted for UTI management and emphasize non-antibiotic approaches for uncomplicated cases. Their attitudes influence counseling practices, highlighting the need for standardized UTI counseling services, antimicrobial stewardship education, and structured communication training to support appropriate antibiotic use. Full article

Among the three lineages of bryophytes, liverworts exhibit a wide variety of terpenoid fingerprints. Terpenoids are abundant in the oil bodies of liverworts. Hornworts and mosses are also reported to contain sesqui-, di-, and triterpenoids, although they lack oil bodies. Overall, the abundance of sesquiterpenoids is much greater than that of other types of terpenoid compounds. The occurrence of triterpenoids is very low. Terpenoids found in higher plants are detected in Marchantiophyta in their enantiomeric forms, with a few exceptions. Organic chemists discovered many di- and sesquiterpenoids with interesting carbon skeletons. Bryophytes possess microbial terpene synthase-like enzymes that are different from typical plant terpene synthases. Original research articles and high-quality reviews were extracted from Google Scholar, PubMed, ScienceDirect, and Scopus using the keywords “terpenoid diversity”, “terpenoid and chemosystematics”, “terpenoids of bryophytes”, “oil bodies”, “terpene synthase”, “microbial terpene synthase-like enzymes”, and “genomics and terpenoid research progress” to prepare this review. Only the literature published in the English language was considered. This review focused on the terpenoid diversity of bryophytes, including chemosystematics uses, as well as the varying carbon skeletons of terpenoids. Oil body biogenesis and evolution, along with the terpene biosynthesis pathway and related enzymes, are briefly covered. The emergent area of multi-omics approaches may bridge the gap that exists in this field and will also open up future avenues for the use of Marchantia polymorpha or Physcomitrella patens as an efficient tool for specific and valuable terpenoid production. Full article

This study investigated the mechanism of TGF-β1-induced Nox4 expression in pulmonary fibrosis (PF) and the anti-fibrotic effects of Tanshinone IIA (Tan-IIA). In a bleomycin-induced pulmonary fibrosis mouse model and in TGF-β1-stimulated fibroblasts, Tan-IIA attenuated fibrosis, oxidative stress, and fibroblast activation. Pharmacological inhibition revealed that the JNK/c-Jun and Smad3 pathways cooperatively mediate TGF-β1-induced expression of Nox4 and fibrotic markers (Collagen I/III, α-SMA). Tan-IIA exerted these effects by dually inhibiting the JNK/c-Jun and Smad2/3 pathways, reducing their phosphorylation and nuclear signaling, which consequently suppressed Nox4 transcription and protein expression. The combination of Tan-IIA with JNK or Smad3 inhibitors synergistically enhanced these effects. We identified a tandem c-Jun/Smad binding element in the Nox4 promoter that is critical for TGF-β1 response. Reporter assays and CUT&RUN experiments confirmed that TGF-β1-induced transcriptional activation depends on an intact c-Jun/Smad binding element and recruitment of c-Jun and Smad2/3. Moreover, Tan-IIA inhibited the enrichment of c-Jun and Smad2/3 at the Nox4 promoter. Collectively, our findings demonstrate that a c-Jun/Smad element integrates profibrotic JNK and Smad signaling to drive Nox4 expression. Tan-IIA presents a novel therapeutic strategy for fibrosis by simultaneously targeting these two key pathways, thereby mitigating Nox4-dependent oxidative stress and fibroblast activation. Full article

Background: Skin aging is driven by oxidative stress and ultraviolet (UV) exposure, leading to extracellular matrix degradation and loss of skin elasticity. This study aimed to identify the most biologically active Citrus species using a bioassay-guided approach and evaluate its potential for dermal applications. Methods: Hydroalcohol extracts and ethyl acetate fractions of Citrus sinensis, Citrus aurantifolia, and Citrus reticulata leaves were screened for antioxidant, enzyme-inhibitory, and polyphenol content. The most active fraction was characterized by UPLC-PDA and LC–MS/MS, formulated into Span-based nanovesicles, and evaluated for physicochemical properties and drug release. Biological activity was assessed using an in vitro scratch wound-healing assay on human dermal fibroblasts and a UVA-induced photoaging mouse model, supported by molecular dynamics simulations. Results: The ethyl acetate fraction of C. aurantifolia (CAE) exhibited the highest biological activity among the tested samples. This fraction showed potent antioxidant activity (DPPH IC₅₀ = 3.53 ± 0.05 µg/mL), marked inhibition of elastase (91.3%), collagenase (92.0%), and tyrosinase (80.2%), and a high total flavonoid content (110.49 mg rutin equivalents/g). Phytochemical profiling of CAE tentatively identified fourteen compounds, predominantly flavonoids, with hesperidin (30.4 mg/g) as a major constituent. The optimized nanovesicles (184 ± 0.9 nm, PDI 0.10, EE% 75.0%) enabled sustained hesperidin release. CAE and CAEnp enhanced fibroblast migration and accelerated wound closure at 24 h (p < 0.05). In vivo, CAEnp improved UVA-induced histopathological alterations and modulated oxidative stress-related markers by reducing p62/SQSTM1 by 28.7%, Keap1 expression to 21% compared with the CAE-treated group, and enhancing Nrf2, ARE, and NQO1 expression by 54.1%, 28.3%, and 57%, respectively. Molecular dynamics simulations supported stable hesperidin binding to elastase and suggested possible modulation of collagenase flexibility. Conclusions: The polyphenol-rich leaf fraction from C. aurantifolia, identified through comparative bioassay-guided fractionation, demonstrated antioxidant, enzyme-inhibitory, wound-healing, and photoprotective effects, particularly after nanoformulation. These findings support its potential for further development as a natural topical anti-aging candidate. Full article

Biocatalysis has emerged as a mainstay in the field of sustainable chemical synthesis owing to its high selectivity, mild reaction conditions, and reduced environmental impact. Traditional enzyme engineering approaches, such as rational design and directed evolution, are often associated with limited throughput and a limited understanding of sequence–structure–function relationships, despite high experimental costs. In recent years, the integration of machine learning (ML) into enzyme engineering has emerged as a transformative approach, enabling data-driven prediction, design, and optimization of biocatalysts, thereby enhancing performance and applications. This review provides a comprehensive overview of ML-guided strategies to improve key enzymatic parameters, including the turnover number (k_cat), substrate affinity (K_m), and catalytic efficiency (k_cat/K_m), with a focus on mechanistic insights and performance outcomes. The integration of ML models into design–build–test–learn (DBTL) cycles accelerated directed evolution, reduced screening efforts, and enabled targeted mutagenesis. Beyond applications, this review also discusses the current limitations of ML-guided approaches, including data scarcity, model interpretability, and challenges in predicting complex mutations and allosteric effects. The gap between computational predictions and experimental outcomes is identified, and the role of ML integration with enzyme kinetics, molecular dynamics, and high-throughput experimentation is emphasized. Future directions, such as generative AI, explainable ML, and autonomous laboratories, are discussed for next-generation biocatalytic applications. Full article

Fritillaria species are rich in steroidal alkaloids and are traditionally used for various medicinal purposes. This study investigated the biological activities of methanol, tertiary alkaloid, and quaternary alkaloid extracts obtained from the bulbs of Fritillaria imperialis and Fritillaria pinardii. Methanol, tertiary alkaloid, and quaternary alkaloid extracts prepared from the bulbs of both species were analyzed for secondary metabolites using LC-HRMS. Antimicrobial, cytotoxic, antioxidant, anti-inflammatory, and anticholinesterase activities were evaluated using standard in vitro assays. LC-HRMS analysis identified fumaric acid as the major metabolite in both species, together with (-)-epigallocatechin, cynarin, herniarin and several pyrrolizidine alkaloid N-oxides. Antibacterial activity was restricted to Staphylococcus aureus, whereas alkaloid-rich fractions displayed strong antifungal activity, particularly against Aspergillus fumigatus and Candida spp., with inhibition zones reaching 40 mm. At 80 μg/mL, the most pronounced reductions in cell viability were observed for F. imperialis methanol extract in A549 cells (55.19 ± 2.74%) and for F. pinardii tertiary alkaloid extract and F. imperialis quaternary alkaloid extract in MCF-7 cells (49.68 ± 1.83% and 49.76 ± 1.09%, respectively), whereas all extracts maintained J774 macrophage viability above 94%. Antioxidant activity was most pronounced in the quaternary alkaloid extract of F. pinardii (DPPH IC₅₀ = 90.42 μg/mL; ABTS IC₅₀ = 57.22 μg/mL), although all extracts were less active than the reference antioxidants. The tertiary alkaloid extract of F. pinardii exhibited potent anti-inflammatory activity, showing 5-LOX inhibition comparable to indomethacin (IC₅₀ = 20.05 ± 0.23 μg/mL) and suppressing LPS-induced nitric oxide production by up to 62.22%. Among the tested samples, the tertiary alkaloid extract of F. pinardii exhibited the strongest cholinesterase inhibitory activity against AChE (IC₅₀ = 13.39 ± 0.02 μg/mL). The results indicate that both F. imperialis and F. pinardii extracts exhibit measurable biological activities under in vitro conditions, with the alkaloid-rich extracts of F. pinardii showing comparatively stronger antifungal, anti-inflammatory, and anticholinesterase effects. Full article

Background/Objectives: Essential Oils (EO), complex mixtures of organic compounds, exhibit a wide range of properties useful in the pharmaceutical, cosmetic, perfumery and agri-food fields. In particular, well-recognized EO anti-inflammatory properties push towards the investigation of the mechanisms underlying them. One of the signaling pathways targeted by EOs is the Janus Kinase Signal of Transducer and Activator of Transcription (JAK/STAT), whose hyperactivation is associated with inflammation, immune diseases and tumor progression. Methods: A comprehensive search on the major bibliographic databases was conducted; current findings and recent insights about the role of EOs in modulating the JAK/STAT Signaling Pathway were collected. Results: EOs derived from different plant species showed efficacy in attenuating the release of pro-inflammatory cytokines and mediators and in inhibiting phosphorylation of both JAK and STAT proteins. These results could be due to the EO’s multi-component nature and to the synergistic interaction occurring within these complex mixtures, both reflecting multi-target effects and modulations. Limitations concerning formulations, lack of standardization, efficacy and safety profiles, sustainable and eco-friendly approaches, and the gap between the literature and translation to the clinic were addressed. Conclusions: EOs represent emergent, promising and high-value candidates able to modulate the JAK/STAT Signaling Pathway. Full article

Gamma irradiation is one of the techniques widely authorized for the decontamination of dried herbs and spices. Its effect on the functional properties of essential oils, however, remains incompletely characterized. In this study, we examined the impact of gamma irradiation (at 5, 15, and 25 kGy) on the phytochemical composition, antimicrobial activity, antioxidant capacity, and insect-repellent activity of Cymbopogon citratus essential oil. The GC-MS analysis revealed that the citral-dominant chemotype remained stable across all irradiation doses, with geranial and neral constituting approximately 62–63% of the volatile profile. The antibacterial assays were done on five bacterial strains (Staphylococcus aureus, Bacillus subtilis, Streptococcus spp., Pseudomonas aeruginosa, and Klebsiella pneumoniae). Inhibition zones showed no statistically significant differences across irradiation doses (p ≥ 0.05), while MIC (75–100 µg/mL) and MBC (125–150 µg/mL) values remained constant across all doses. DPPH, ABTS, and FRAP antioxidant assays revealed no dose-dependent changes (DPPH IC₅₀: 688–703 µg/mL; ABTS IC₅₀: 18–22 µg/mL; FRAP: 505–517 µg/mL ascorbic-acid equivalents). The essential oil exhibited pronounced repellent activity (87–99%) against adult Tribolium confusum beetles at 0.125 µL/cm², persisting for 24 h and unaffected by irradiation. Molecular docking of the major constituents (geranial, neral, geraniol, and β-myrcene) against key target proteins (3N7H, 3NVY, 4URM, and 8BN6) provided predictive support consistent with the observed activities, indicating plausible molecular interactions rather than confirmed target engagement. In silico ADME and toxicity profiling indicated favorable predicted pharmacokinetic properties and no major in silico toxicity alerts for the four modeled constituents. Taken together, these findings indicate that, under the conditions tested, gamma irradiation at food-decontamination doses produced no major shifts in composition and no statistically detectable changes in the measured bioactivities of C. citratus essential oil. Full article

Background: Antibody–drug conjugates (ADCs) have evolved from early heterogeneous constructs into a mature therapeutic platform with exponential clinical relevance. This review highlights recent advances in ADC design and development, with emphasis on antigen selection, antibody engineering, linker and payload innovation, site-specific conjugation, clinical translation, toxicity, resistance, and emerging data-driven approaches. Methods: The review draws on the literature published from 2019 to the recent clinical and regulatory developments relevant to approved and late-stage ADCs, emphasizing the advances in target biology, antibody formats, linker chemistry, payload classes, conjugation technologies, developability assessment, and computational or artificial intelligence-assisted design strategies. Results: ADC development has evolved with improved target selection, enhanced internalization and tumor selectivity, and the use of engineered, bispecific, biparatopic, and fragment-based antibody formats. Linker and payload innovation has expanded beyond traditional microtubule inhibitors to include topoisomerase I inhibitors, DNA-damaging agents, and emerging dual-payload or non-cytotoxic strategies. Site-specific conjugation and improved control of drug-to-antibody ratio have increased stability, pharmacokinetic performance, and manufacturability. Clinically, ADCs are being used across a broader range of malignancies and treatment settings, although toxicities and resistance mechanisms remain an important limitations. Computational methods and artificial intelligence are increasingly being explored for target discovery, molecular optimization, toxicity prediction, and model-informed clinical development. Conclusions: ADCs are transitioning toward a more integrated, design-driven platform in which antigen biology, antibody format, chemistry, and computational prediction are jointly optimized. Future progress will depend on improved standardization, biomarker-guided development, and interdisciplinary approaches to enhance its therapeutic index and expand its applications beyond oncology. Full article

This study highlights the therapeutic innovation potential of bioactive plant extracts derived from the selected Thai Rejuvenation Remedy 2 (TRJ 2) for antidiabetic applications. By integrating phytochemical profiling with in vitro α-glucosidase inhibition assays and in silico analyses, including molecular docking and density functional theory (DFT), a comprehensive evaluation of the extract’s bioactivity and mechanistic basis was achieved. The findings demonstrate that both the relative abundance and chemical reactivity of constituent compounds contribute to the overall inhibitory effect through synergistic and competitive interactions. Piperine was identified as a potential bioactive metabolite, with a theoretically strong binding affinity and high reactivity toward the target enzyme. Among the tested extracts, the 80% ethanol extract exhibited the highest inhibitory activity (IC₅₀ = 34.32 µg/mL), underscoring the importance of extraction optimization for maximizing therapeutic efficacy. Furthermore, formulation of the extract with cyclodextrin significantly enhanced solubility and improved pharmaceutical characteristics, meeting the quality requirements of the Thai Herbal Pharmacopoeia (THP). However, the detection of cadmium slightly above permissible levels indicates the need for stricter raw material quality control. Overall, TRJ 2 represents a promising source of bioactive plant-derived compounds and a viable candidate for development into innovative, ready-to-use herbal therapeutics for diabetes management. Full article

Acne is a common inflammatory skin condition that significantly impacts quality of life. Standard treatments often cause skin irritation or contribute to antibiotic resistance. Cannabidiol (CBD) has demonstrated anti-inflammatory and sebum-regulating properties; however, its application is limited by poor solubility and stability. This study investigated the physicochemical properties of a CBD-loaded hyaluronic acid–graft-poly(N-isopropylacrylamide) nanogel (Hy-CBD). The biological activities of Hy-CBD, including its anti-inflammatory and antioxidant effects, were also evaluated. In addition, an exploratory clinical study was conducted to assess the safety and efficacy of the formulation in 22 Asian participants with inflammatory acne. In this open-label, non-randomized study, participants applied the gel twice daily for seven days. Assessments of skin tolerance, lesion size, redness, and pigmentation were performed at baseline, Day 2, and Day 7 using clinical examination and imaging analysis. The Hy-CBD gel was clinically tolerated, with no evidence of comedogenic or acnegenic potential. By Day 7, inflammatory lesion size was reduced by 46%, with significant improvements in redness and post-inflammatory pigmentation. All participants reported a subjective reduction in acne severity and expressed satisfaction with the treatment outcomes. These findings suggest that the Hy-CBD gel is a safe and promising delivery system for acne management. Nevertheless, larger randomized controlled studies are required to validate these preliminary findings. Full article

Background: Mitophagy is a mitochondrial quality-control pathway whose contribution to cancer stress tolerance may vary by cellular context. For essential oils, mechanistic interpretation is often limited by compositional variability and the limited number of studies addressing malignant and non-malignant comparisons under matched exposure conditions. Methods: Ylang Ylang essential oil (YY EO) was characterized by GC-MS-FID. Lung cancer cells (A549) and a salivary gland carcinoma model (HTB-41), together with non-malignant lung-related cells (BEAS-2B, MRC-5), were exposed to YY EO. Functional outcomes were assessed by WST-1 and LDH assays. Mitophagy-related and mitochondrial quality-control-associated genes were quantified by RT-qPCR (2^−ΔΔCt). Results: GC-MS-FID identified a terpenoid-rich mixture (99.31%), with germacrene D and β-caryophyllene among the major constituents. YY EO was associated with dose- and cell-type-dependent functional responses, with malignant cells showing reductions in WST-1 signal and stronger LDH-associated responses under the tested conditions, while non-malignant cells showed less pronounced functional changes. Transcriptional responses were context-dependent, with differential changes in mitophagy-related genes across cell lines. Conclusions: These findings provide comparative evidence of greater functional sensitivity in malignant cells, alongside cell-context-dependent mitophagy-related transcriptional responses. These observations are hypothesis-generating and remain limited to functional readouts and mRNA-level data. Within these limits, the present study provides a composition-anchored comparative dataset that may support future mechanistic studies in this area. Full article