Search Results (14,624)

Background: Rising antifungal and antibiotic resistance among Candida species, Staphylococcus aureus, and Enterococcus faecalis has renewed interest in antimicrobial photodynamic therapy (aPDT) as a resistance-independent strategy. Riboflavin 5′-phosphate is a biocompatible, blue-light-activated photosensitizer, but standardized dosing across fungal and bacterial targets is lacking. Objective: The aim of this study was to systematically optimize pre-irradiation incubation time, photosensitizer volume, irradiation time, and laser power for riboflavin 5′-phosphate aPDT (450 nm diode laser) against C. albicans, C. glabrata, C. krusei, S. aureus, and E. faecalis, and compare species susceptibility under optimized conditions. Methods: ATCC strains were treated with 0.1% riboflavin 5′-phosphate across four groups (photodynamic, photosensitizer-only, laser-only, control) in a staged design optimizing incubation (1–30 min), photosensitizer volume (50–150 µL), irradiation time (10–120 s), and power (50–400 mW). Viable counts (CFU/mL) were quantified. Results: Significant reductions occurred only with combined light-plus-photosensitizer treatment. Optimal parameters were 15 min incubation, 100 µL photosensitizer for Candida spp. (50 µL for bacteria), and 120 s at 400 mW, though C. albicans and C. krusei plateaued by 60 s. Maximum reductions were modest: 53.5% (C. albicans), 46.7% (S. aureus), 37.9% (C. glabrata), 35.9% (C. krusei), and 26.5% (E. faecalis), all below 1 log₁₀. A significant light × photosensitizer interaction, confirming photodynamic specificity, was seen for C. albicans, C. glabrata, and S. aureus, but not C. krusei or E. faecalis. Conclusions: Riboflavin 5′-phosphate aPDT under 450 nm light produces reproducible, dose-dependent, species-specific antimicrobial activity, best suited as an adjunctive rather than stand-alone therapy pending biofilm and in vivo validation. Full article

Background: The solute carrier family 22 member 23 (Slc22a23), a gene encoding a membrane-orphan transporter, is irreversibly upregulated in the brains of rats following fetal undernutrition. Materials and Method: To identify potential substrates of the SLC22A23 transporter, we analyzed the plasma of both Slc22a23-proficient and deficient rats. Results: We found that the level of lysophosphatidylcholine a C20:4 (LPC(20:4)) in plasma was more than twofold lower in the Slc22a23^−/− rats than in the Slc22a23^+/+ rats. We then conducted a series of animal behavioral tests on Slc22a23-proficient (Slc22a23^+/+) and deficient (Slc22a23^−/−) rats to investigate the effects of LPC(20:4) administration. We observed considerable effects in both Slc22a23^+/+ and Slc22a23^−/− rats, with more pronounced effects in the Slc22a23^+/+ rats. These effects included reductions in total distance travelled in an open field test, reductions in access to the novel objects in a novel object recognition test, reductions in sociability to a familiar rat in a social interaction test, and shorter latencies to the target in the Morris water maze test. Discussion: These findings suggest that LPC(20:4) administration enhances memory acquisition in both Slc22a23^+/+ and Slc22a23^−/− rats, though it appears to be more effective in the Slc22a23^+/+ rats. While the SLC22A23 transporter is likely involved in facilitating the transport of LPC(20:4), other transporters may also play a role in its transport in vivo. Full article

Background/Objectives: Resveratrol (RSV) is a phenolic compound that possesses antioxidant properties and whose biological application has been limited by its physicochemical properties, including its low solubility and stability. Methods: In this study, polymeric nanoparticles (NPs) were developed using polylactic acid (PLA) to encapsulate resveratrol and improve its bioavailability, using a factorial design to optimize the formulation and evaluate its cytotoxicity against breast cancer cells. Results: The optimized system presented an average size of 389.2 nm, a PDI of 0.127, a negative z-potential, and an encapsulation efficiency of 76.02%. The statistical analysis indicated that the PLA: RSV ratio was the main determinant of encapsulation percentage and particle size (p < 0.05). On the other hand, with the hemolysis test, blood compatibility was evidenced at all concentrations evaluated, while in the cell assays, the PLA-RSV nanoparticles significantly decreased the cellular viability of MCF-7 and MDA-MB 231 cells compared to untreated cells, and in combination with NP PLA-RSV and doxorubicin, greater cytotoxicity was produced in MCF-7 cells compared to nanoparticle-only treatments (p < 0.05, ANOVA, via followed by a Fisher test). Conclusions: In addition, distinct cytotoxic effects are observed, associated with their tumor phenotype. Taken together, these results suggest that PLA-RSV NPs are a useful system for future studies on redox status and other biological processes important in cancer. Full article

Background/Objectives: Dry eye disease (DED) is a multifactorial disorder characterized by tear film instability, inflammation, and ocular surface damage, which significantly impairs visual function and quality of life. Conventional ophthalmic formulations, such as eye drops, have low bioavailability owing to rapid elimination, necessitating frequent administration. In this study, we developed an eyelid-applied drug delivery system (DDS) based on troxipide (TRO) nanoparticle formulation (TRO-NP@EG) to achieve sustained ocular surface delivery. Methods: TRO nanosuspensions were prepared by wet bead milling and incorporated into a Carbopol-based gel. Particle size, dispersion stability, and uniformity were evaluated, and in vitro drug release studies was compared with that of TRO-MP@EG. In vivo drug transfer into tear fluid was assessed in rabbits following eyelid application, and therapeutic efficacy was evaluated in an N-acetylcysteine-induced dry eye model. Results: TRO nanosuspensions had a mean particle size of approximately 118 nm. TRO-NP@EG exhibited superior dispersion stability and uniformity and achieved 2.5-fold higher drug release than TRO-MP@EG, while the nanoparticles remained in solid form. In vivo studies in rabbits, TRO-NP@EG significantly enhanced drug transfer into tear fluid, primarily via the meibum pathway. Furthermore, TRO-NP@EG significantly improved mucin levels, tear secretion, and tear film stability compared with TRO-MP@EG in an N-acetylcysteine-induced dry eye model. Conclusions: These findings suggest that eyelid application of nanoparticle-based formulations enables efficient and sustained drug delivery to the ocular surface via the meibomian glands. Therefore, TRO-NP@EG represents a promising therapeutic strategy for DED, providing enhanced efficacy and a novel route of administration for ophthalmic DDSs. Full article

Background/Objectives: Skin cancer is a major health concern worldwide and has led to the quest for safer and more effective topical chemopreventive agents. In this study, the in vivo efficacy of a hydrogel formulation containing kaempferol and ferulic acid-loaded nanoparticles was evaluated for the management of DMBA-croton oil-induced skin cancer in Swiss albino mice. Methods: Drug-loaded nanoparticles prepared via nanoprecipitation were incorporated into hyaluronic acid to obtain single-drug (FAPG, KMPG) and dual-drug-loaded (FKMG) hydrogel formulations. Skin tumors were induced via DMBA as an initiator followed by croton oil as a promoter, with tumor onset observed after a similar latency period of approximately 5–6 weeks in all carcinogen-exposed groups. Treatment was initiated after week 6 and continued until week 16. Results: The gel formulation had a skin-compatible pH and the desired rheological and spreadability properties. The dual drug-loaded hydrogel formulation had a more controlled and prolonged release profile. Compared with the negative and vehicle control groups, the FKMG-treated group presented a marked reduction in tumor incidence and tumor burden, along with improved body weight gain. Biochemical analysis revealed the restoration of antioxidant and enzyme activity in treated animals, particularly in animals treated with FKMG. Histopathological examination revealed near-normal skin architecture in FKMG-treated mice, indicating strong protective effects. Additionally, significant downregulation of TNF-α and IL-6 suggested effective suppression of tumor-promoting inflammatory pathways. Conclusions: Overall, the FKMG formulation exhibited superior chemopreventive efficacy compared with the FAPG and KMPG treatments, highlighting its potential as a promising topical therapeutic strategy against chemically induced skin carcinogenesis. Full article

Background/Objectives: Meropenem is a broad-spectrum carbapenem antibiotic with demonstrated efficacy against multidrug-resistant Gram-negative pathogens. Although its use as an ophthalmic formulation is off-label, growing clinical evidence supports its application in severe ocular infections such as keratitis and endophthalmitis. However, the intrinsic instability of meropenem in aqueous solutions and the absence of standardized ophthalmic preparations limit its routine use. Furthermore, no stability studies are currently available for meropenem 50 mg/mL eye drops stored in polypropylene (PP) dropper bottles under freezing and subsequent refrigerated conditions. The aim of this study was to evaluate the physicochemical and microbiological stability of a 50 mg/mL meropenem ophthalmic solution prepared in a hospital pharmacy using a commercial meropenem pharmaceutical product, and packaged in PP containers. Methods: Eye drops were aseptically prepared from a commercially available pharmaceutical product, containing 1g de meropenem and anhydrous sodium carbonate as an excipient. After preparation, the drops were stored at −20 ± 2 °C for up to 42 days, followed by refrigerated storage (5 ± 3 °C) after thawing for up to 7 days. Chemical stability was assessed using a validated stability-indicating HPLC method in accordance with ICH guidelines and was defined as 90–110% recovery of the initial concentration. Physical stability (appearance, pH, particulate matter) and microbiological stability were also evaluated under simulated in-use conditions. Results: The HPLC method demonstrated excellent linearity, precision, and accuracy. Meropenem concentrations remained within the predefined acceptance limits throughout the 42-day study period under freezing conditions, with no significant changes in pH, color, or particulate formation. After thawing, a progressive decrease in drug concentration was observed under refrigerated conditions, falling below 90% of the initial concentration within 24–48 h. A concomitant color change from colorless to yellow was also detected, consistent with β-lactam ring hydrolysis. Despite this degradation, no significant changes in physical parameters other than color were observed, and microbiological testing confirmed sterility for up to 7 days under refrigerated conditions. Conclusions: Meropenem drops 50 mg/mL in PP dropper bottles are physicochemically and microbiologically stable for 43 days (42 days under frozen conditions plus 1 day, in-use conditions, after opening and under refrigeration). Full article

T-cell immunoreceptor with immunoglobulin and immunoreceptor tyrosine-based inhibitory motif (ITIM) domains (TIGIT) emerged as one of the most promising next-generation immune checkpoint targets following the success of programmed cell death protein 1 (PD-1), programmed death-ligand 1 (PD-L1), and cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) blockade. TIGIT suppresses antitumor immunity through interaction with cluster of differentiation 155 (CD155), inhibition of CD226-mediated co-stimulation, and promotion of immunosuppressive regulatory T-cell (Treg) activity within the tumor microenvironment (TME). Strong preclinical evidence demonstrated that TIGIT blockade, particularly in combination with PD-1/PD-L1 inhibition, restored T-cell and natural killer (NK) cell function and produced durable antitumor responses in multiple tumor models, leading to rapid clinical development. Despite this compelling biological rationale, most late-stage clinical programs failed to reproduce early success. Although the phase II CITYSCAPE trial showed encouraging activity in PD-L1-high non-small cell lung cancer (NSCLC), subsequent phase III trials, including SKYSCRAPER-01, SKYSCRAPER-02, SKYSCRAPER-03, SKYSCRAPER-14, AdvanTIG-302, KEYVIBE, and STAR-221, failed to improve survival outcomes or meet primary endpoints. The notable exception was SKYSCRAPER-08 in esophageal squamous cell carcinoma, suggesting that TIGIT blockade may be effective only in selected biological contexts. This review critically examines the molecular biology of the TIGIT–CD155–CD226 axis, its role in immune regulation and tumor immune evasion, and the preclinical and clinical evidence supporting TIGIT-targeted therapy. Particular emphasis is placed on understanding the causes of clinical failure, including CD226 loss during T-cell exhaustion, checkpoint network redundancy, Fc-engineering uncertainty, immunosuppressive TMEs, inadequate biomarker-guided patient selection, and tumor-type-specific dependence on the TIGIT pathway. We also present original bioinformatics analyses demonstrating that broader checkpoint network signatures outperform TIGIT expression alone for patient stratification. Finally, we evaluate emerging solutions including biomarker-guided precision immunotherapy, Fc-optimized antibodies, bispecific checkpoint inhibitors, TIGIT-engineered chimeric antigen receptor T-cell (CAR-T) cells, radiotherapy combinations, and multi-checkpoint blockade. Collectively, current evidence suggests that the future of TIGIT-directed therapy lies not in universal checkpoint inhibition but in biologically informed, precision-guided immunotherapy strategies. Full article

Background: Substantial inter-individual variability in imatinib systemic exposure may influence both efficacy and toxicity. Based on recommendations of the International Association for Therapeutic Drug Monitoring and Clinical Toxicology (IATDMCT), the IRCCS-CRO of Aviano started in 2018 offering physicians the opportunity to monitor plasma concentrations of imatinib and its active metabolite, norimatinib, as part of a diagnostic service for patients with Gastrointestinal Stromal Tumor (GIST). This study aims to evaluate the potential clinical utility of imatinib Therapeutic Drug Monitoring (TDM) in predicting response and toxicity in a real-world cohort of 63 patients with GIST. Methods: Imatinib and its active metabolite norimatinib trough concentrations (C_min) were quantified using a validated LC–MS/MS method. Associations between drug exposure and clinical–demographic characteristics, treatment-related toxicity, and progression-free survival (PFS) were evaluated. Results: A total of 437 plasma samples from 63 patients were collected. Marked inter- and intra-patient variability in imatinib exposure was observed (43% and 31% respectively); 67% of patients receiving 400 mg/day had a C_min below the recommended target concentration (1100 ng/mL). Female sex was significantly associated with higher imatinib exposure (median C_min 1114 vs. 786 ng/mL in males; p = 0.0018). Combined age–sex analysis showed a significant difference between women and men < 60 years (1041 vs. 668 ng/mL; p = 0.0068, Bonferroni-adjusted). A strong exposure–toxicity relationship emerged, with higher imatinib levels in samples collected at toxicity occurrence vs. the others (1728 vs. 927 ng/mL; p < 0.0001), without a direct association between sex and toxicity. No significant association between C_min and PFS was observed; however, disease progression was more frequent in patients with C_min < 500 ng/mL and absent in those with C_min >1500 ng/mL (60% vs. 0%, respectively). Conclusions: TDM was found to be a potentially useful tool for predicting clinically relevant toxicity and outcome. Sex and age significantly influence exposure, supporting tailored dosing strategies. Full article

Background/Objectives: Epilepsy remains a major neurological disorder, with approximately one-third of patients continuing to experience pharmacoresistant seizures despite the availability of numerous antiseizure medications (ASMs). While current therapies primarily target neuronal hyperexcitability through modulation of ion channels and neurotransmitter systems, increasing evidence suggests that epileptogenesis arises from multiscale interactions involving molecular, cellular, circuit, network, neuroinflammatory, and neurovascular mechanisms. Although therapeutic strategies have diversified, this expanded mechanistic understanding has not yet been fully incorporated into therapeutic development and evaluation. This review integrates current knowledge of multiscale epilepsy pathophysiology with recent therapeutic advances and emerging experimental platforms. Methods: This narrative review synthesized literature identified primarily through PubMed and Google Scholar searches through January 2026, supplemented by targeted updates of therapeutic development and regulatory status through July 2026. Particular emphasis was placed on ion channel modulators, synaptic and neuromodulatory therapies, neuroinflammatory interventions, precision genetic approaches, and human-relevant experimental platforms, including induced pluripotent stem cell (iPSC)-derived models, brain organoids, multi-electrode arrays (MEAs), organ-on-a-chip systems, multi-omics technologies, and artificial intelligence (AI)-based analytical frameworks. Results: Current and emerging therapies target increasingly diverse molecular, circuit, neuromodulatory, and neuroinflammatory mechanisms. However, drug resistance remains multifactorial, and the long-term effects of therapeutic interventions on network remodeling, neuro-glial interactions, and sustained clinical response remain incompletely understood. NAMs provide complementary capabilities for patient-specific disease modeling, functional network phenotyping, neurovascular modeling, and the integration of molecular, electrophysiological, and computational data across biological scales. Conclusions: Epilepsy is increasingly recognized as a multiscale network disorder rather than solely a condition of neuronal hyperexcitability. The coordinated use of complementary human-relevant platforms may help incorporate multiscale mechanistic insights into therapeutic development and evaluation, narrow persistent translational gaps, and support more predictive and mechanism-informed treatment strategies. Full article

The inappropriate utilization of traditional antimicrobials has expedited the development of multidrug-resistant bacterial strains, thereby generating an urgent demand for innovative, safe, and efficacious alternatives. Graphene oxide (GO), characterized by its distinctive physicochemical properties, holds significant potential for a wide range of biomedical applications, such as the prevention of oral diseases, periodontal therapy, control of gastrointestinal infections, wound healing, antibacterial therapy for the respiratory tract, and management of urogenital infections. The antimicrobial mechanisms of GO include physical disruption of microbial membranes, induction of reactive oxygen species (ROS), and its role as a photosensitizer. Moreover, improving the dispersion, stability, release behavior, and reusability of GO-based composites can markedly enhance their effectiveness. The antimicrobial activity of GO is further modulated by its physicochemical properties, synthesis methods, bacterial characteristics, and environmental conditions, while potential synergistic or antagonistic interactions remain to be elucidated. This review synthesizes recent advancements in GO-based antimicrobial strategies, elucidates key mechanisms and influencing factors, and addresses major challenges and future research directions, with the objective of facilitating the clinical translation of GO as a next-generation antimicrobial material. Full article

Background/Objectives: Rules for predicting blood–brain barrier (BBB) permeability, including the CNS multiparameter optimization (CNS MPO) score, Lipinski’s Rule of Five, and Veber’s rules, were developed using relatively limited datasets and have not been systematically re-evaluated using modern large-scale experimental databases. Using the B3DB database, which contains 1058 experimentally measured logBB values, we derived quantitative, data-driven structural thresholds for BBB permeability and benchmarked them against established heuristic rules. Methods: Six key physicochemical properties were calculated for all compounds, and optimal classification thresholds were identified through exhaustive optimizations. Decision trees and scaffold analyses were used to generate interpretable medicinal chemistry guidelines. Results: The topological polar surface area (TPSA) emerged as the strongest single predictor of BBB permeability, with an optimal threshold of 66.8 Ų (AUC = 0.731, 95% CI: 0.689–0.771). This threshold outperformed the approximated CNS MPO ≥ 4 (AUC = 0.625), Lipinski’s Rule of Five (AUC = 0.546), and Veber rules (AUC = 0.566). A simple two-parameter rule combining TPSA < 67 Ų and H-bond donors ≤ 1 achieved 96.6% precision for BBB-permeable compounds while maintaining an AUC of 0.720. Decision tree analysis further confirmed TPSA as the dominant determinant of BBB permeability, whereas scaffold analysis identified the molecular frameworks associated with highly permeable and impermeable compounds. External validation provided preliminary support for the improved specificity of the proposed rule, compared with existing approaches. Conclusions: These findings suggest that the commonly applied TPSA threshold of 90 Ų may be overly lenient. A data-driven threshold of approximately 67 Ų substantially improved the discrimination of BBB permeability across the entire dataset. Compounds with TPSA values between 67 and 90 Ų should be assessed on a case-by-case basis, considering the ionization state and active transport potential, rather than being automatically classified as BBB-permeable. These experimentally grounded rules offer a practical framework for the early-stage design of CNS leads. Full article

Background/Objectives: Olive leaf polyphenols exhibit strong antioxidant and antidiabetic potential. However, their application in nutraceutical and pharmaceutical products is hindered by limited stability, poor solubility, and susceptibility to gastrointestinal degradation. Liposomes offer a viable strategy to enhance their protection and functional performance. Methods: A formulation-driven strategy was employed to develop a gastro-resistant liposomal system for olive leaf polyphenols. Extraction conditions were optimized using a Box–Behnken response surface design to maximize phenolic recovery while ensuring compatibility with phospholipid-based systems. Antioxidant activity and key secoiridoids were assessed by spectrophotometric assays and LC–MS/MS. The optimized extract was incorporated into uncoated and Eudragit^® L100-coated liposomes, which were physicochemically characterized. Antidiabetic effects were evaluated in intestinal STC-1 cells under glucose-induced hyperglycemic conditions. Results: The optimized extract (OE) exhibited high antioxidant activity (124.56 ± 9.57 mg GAE/g, 151.61 ± 2.77 mg TE/g, and 472.92 ± 26.14 mg TE/g in TPC, DPPH and FRAP assays, respectively). LC-HRMS metabolomic profiling confirmed a balanced phytochemical composition, with oleuropein as the main compound (143.144 ± 4.914 mg/g). The optimized Eudragit^®-coated liposomes were spherical unilamellar vesicles with a mean diameter of 101 ± 6.1 nm, moderate polydispersity (0.46 ± 0.02), and a negative zeta potential (−17 ± 3.5 mV). High entrapment efficiency was achieved, reaching 65 ± 2.1% for oleuropein and 96 ± 0.3% for hydroxytyrosol. The structural integrity of the vesicles was maintained during storage and in the simulated gastrointestinal environment. The nanoformulation reduced intestinal glucose uptake and intracellular reactive oxygen species levels, and restored GLP-1 levels. Conclusions: The combination of optimized extraction and liposome-based formulation enabled the development of stable, delivery-ready olive leaf polyphenols for potential nutraceutical and pharmaceutical applications. Full article

Background: Hepatocellular carcinoma (HCC) is a highly lethal malignancy and remains a major contributor to global cancer mortality. In situ vaccination (ISV) holds great potential for HCC therapy. Still, its efficacy is severely limited by intrinsic antigen scarcity and the tumor-associated macrophage (TAM)-mediated inhibition of dendritic cell (DC) maturation and T cell activation. In particular, the M2 TAM-HCC cell crosstalk may further aggravate ISV suppression. Moreover, as an internal organ tumor, HCC requires systemic administration of ISV agents, which often cause severe off-target toxicity. Methods: To address these challenges, we developed secretory phospholipase A2 (sPLA₂)-responsive liposomes co-loaded with the TLR7/8 agonist R848 and the ICD inducer doxorubicin (DOX) via a sequential remote loading method, termed sP-Lips@DR. Results: Incorporation of cholesterol (CHOL) significantly improved DOX encapsulation, and the sequential loading method enabled efficient co-encapsulation of both R848 and DOX. sP-Lips@DR responds to an sPLA₂-overexpressed, mildly acidic microenvironment in HCC, enabling selective drug release. In addition, sP-Lips@DR exhibited sPLA₂-responsive cytotoxicity toward H22 cells and macrophage phenotypic shift. Furthermore, sP-Lips@DR could disrupt the crosstalk between M2-like macrophages and H22 cells in an sPLA₂-responsive manner. Conclusions: Overall, the preparation and in vitro evaluation of sP-Lips@DR demonstrate their potential to enhance ISV efficacy in HCC, providing a solid foundation for future in vivo investigations. Full article

Background/Objectives: Glioblastoma (GBM) remains one of the most aggressive primary brain malignancies, with limited therapeutic progress over the past two decades. Systemic administration of temozolomide (TMZ) is a pillar of clinical management but is constrained by poor brain penetration, short half-life, and systemic toxicity. Localized drug delivery systems represent a compelling approach to address these limitations. We report the development and evaluation of an injectable poly(sebacic acid–ricinoleic acid) poly(anhydride-ester) (pSARA) gel for sustained intratumoral delivery of TMZ. Methods: The pSARA gel was synthesized using a one-pot melt polycondensation technique, and its in vitro release dynamics were assessed using spectrophotometry. In vivo efficacy of the TMZ-loaded pSARA gel was evaluated as a monotherapy and as an adjuvant to radiation or surgical resection using an orthotopic 9L gliosarcoma rat model. Results: The formulation exhibits shear-thinning behavior, enabling syringe-based administration, and undergoes surface erosion in aqueous environments to achieve controlled drug release. In vivo, the TMZ-loaded pSARA significantly prolonged survival compared to controls and outperformed paclitaxel-loaded formulations. Furthermore, combination therapy with radiation or surgical resection demonstrated combined survival benefits, including long-term survivors. Conclusions: These findings highlight the translational potential of pSARA-based local delivery systems as an adjunct or alternative to systemic chemotherapy in GBM treatment. Full article