Hormones regulate many essential biological processes by interacting with specific receptors that control gene expression, metabolism, growth, and immune function. Because numerous therapeutic compounds can influence or disrupt hormone signaling pathways, understanding drug-hormone receptor interactions (DHRI) is crucial for ensuring both therapeutic efficacy and endocrine safety. However, computational approaches for predicting DHRI remain limited, and most existing models do not explicitly incorporate hormone receptor-specific information. In this study, we propose a receptor-aware deep learning framework for DHRI prediction that integrates structural drug features with contextually embedded hormone receptor information. Drug molecules are represented using a hybrid encoding strategy that combines Morgan fingerprints and graph transformer-based molecular features to capture both global chemical properties and local structural information, while hormone receptor sequences are encoded using the pretrained ESM2 protein language model to obtain biologically meaningful sequence representations. The fused drug-hormone receptor features are then processed through a multilayer neural network to predict interaction probabilities. Model performance was evaluated using three splitting strategies, including random split, cold-drug split, and scaffold-based split, to assess both predictive accuracy and generalization ability. Under the random split setting, the model achieved an accuracy of 0.93, sensitivity of 0.94, specificity and precision of 0.92, F1-score of 0.93, and MCC of 0.86 on an independent data set, while also maintaining comparable performance under the more stringent cold-drug and scaffold-based settings. Feature importance analysis showed that atomic identity, hybridization, atom degree, and bond type were key shared determinants, while receptor-stratified results revealed receptor-dependent contributions from secondary features such as aromaticity, chirality, etc. In addition, t-SNE visualization showed clear class separation after training, and molecular docking across estrogen, androgen, and glucocorticoid receptors further supported the biological relevance of the predictions. Together, these findings demonstrate that incorporating hormone receptor-specific sequence information enables more reliable and biologically meaningful prediction of drug-hormone receptor interactions.
Genomic and proteomic initiatives have accelerated therapeutic target identification, yet many proteins remain difficult to drug, especially those lacking structural information, including those with intrinsically disordered regions. However, the conformational states of these proteins are shaped in living cells by chemical modifications, protein-protein interactions, oligomerization, and aggregation, offering new ligandable opportunities. In this article, we first discuss current approaches toward drugging the 'undruggable' proteins, in particular the advantages and limitations of structure-based drug design. We then describe the advantages of cellular protein-based strategies in identifying ligands for proteins that are hard to target, highlighting large-scale chemoproteomics approaches for drugging the proteome.
Niosomes are versatile nanocarriers capable of enabling targeted delivery and controlled release of anticancer agents. Incorporation of ionic surfactants offers an effective strategy to functionalize these systems. In particular, amino acid-derived gemini catanionic niosomes, composed of multifunctional surfactants, represent a promising platform for advanced drug delivery applications. An integrated computational framework combining molecular modeling with design of experiments (DoE) was employed to investigate key properties of amino acid-based gemini catanionic niosomal bilayer. An arginine-derived gemini surfactant was selected as the cationic component, while sodium laurate was used as the anionic surfactant. Two clinically approved anticancer drugs with distinct intracellular targets, niraparib and lapatinib, were evaluated. The findings demonstrate that the bilayer composition and structure strongly influence drug transport across the niosomal membrane. The ratio of diffusion coefficients of the two drugs was identified as a critical performance metric. Among the evaluated formulations, F7 exhibited the most favorable ratio (5.46). Numerical optimization of the diffusion responses yielded an optimized formulation with an improved ratio of 10.28. The corresponding diffusion coefficients for lapatinib and niraparib were 8.52 × 10- 12 and 8.28 × 10- 13 m²/s, respectively. Lapatinib diffusion was enhanced through synergistic interactions between surfactant components, whereas niraparib diffusion followed an additive trend predominantly governed by sodium laurate concentration. These results highlight the capability of molecular modeling integrated with experimental design to guide the rational optimization of nanocarrier systems. Moreover, amino acid-based gemini catanionic niosomal bilayer offer tunable properties that can be exploited for controlled and site-specific delivery of anticancer drug combinations.
Indonesia has one of the highest number of tuberculosis (TB) cases globally. The previous treatment policy of the Ministry of Health was to administer TB drugs intermittently (three times per week) during the continuation phase. Since 2023, the treatment policy has changed to daily dosing during the continuation phase. However, evidence comparing the treatment outcomes and tolerability of these agents remains limited. This study compared patient characteristics, treatment success, and reported adverse events between intermittent and daily regimens among drug-susceptible TB patients. An observational cohort study was conducted using secondary data from medical records and the National TB Information System, with prospective ascertainment of adverse events via standardized telephone and face-to-face interviews among a subset of participants. Group comparisons were performed using chi-square tests, t-tests, and multivariable logistic regression (adjusted for age, HIV status, diabetes status, and baseline sputum). A total of 532 drug-susceptible TB patients were included (intermittent n = 247; daily n = 285). The daily group had a higher mean age and a greater proportion of HIV-positive and diabetic patients (p < 0.05). Treatment success rates were comparable between the two groups (87.85% vs. 87.37%; p = 0.850), with no significant association observed in the adjusted analyses (aOR = 1.23 [0.69-2.18]; p = 0.494). Among 327 patients with adverse event data available (61.47%), reported adverse events were more frequent in the daily group (100.00% vs. 84.38%; p < 0.001), particularly nausea/vomiting/fatigue/fever (aOR = 3.03; 95% CI: 1.69-5.55) and itching (aOR = 2.07; 95% CI: 1.27-3.41); however, these findings were based on a subset of participants and may be subject to recall and reporting bias. Intermittent and daily continuation-phase regimens showed comparable treatment success in this observational study. Among participants with available adverse event data, daily dosing was associated with more frequently reported adverse events; however, causal inference could not be made due to non-random regimen allocation, baseline differences between groups, incomplete ascertainment of adverse events, and potential recall and reporting bias. These findings suggest the potential importance of routine tolerability monitoring and targeted patient support in programmatic TB care, though confirmation from multicenter prospective studies is needed given the single-center design and incomplete adverse event ascertainment. Clinical trial number: not applicable.
Drug resistance remains a major challenge in the treatment of ovarian cancer and is a leading cause of recurrence and poor clinical outcomes. Increasing evidence has identified acetylation as a critical epigenetic and post-translational modification that contributes to therapeutic resistance through the regulation of chromatin accessibility, gene transcription, protein function, and cellular signaling. Recent studies suggest that acetylation functions as an integrative regulatory hub linking multiple resistance-associated processes, including DNA damage repair, cancer stemness, cellular plasticity, metabolic adaptation, and immune evasion. In this review, we summarize the molecular mechanisms by which histone and non-histone acetylation, together with acetylation-associated regulatory networks, drive ovarian cancer drug resistance. We further discuss current advances in acetylation-targeted therapeutic strategies, including HDAC inhibitors, BET inhibitors, emerging KAT inhibitors, and rational combination approaches designed to overcome resistance. Collectively, acetylation-centered regulatory networks represent promising therapeutic targets in ovarian cancer. A deeper understanding of these mechanisms may facilitate biomarker-guided patient stratification and support the development of more effective strategies for overcoming drug resistance.
Peroxynitrite (ONOO-) plays a pivotal role in the pathogenesis of Rheumatoid arthritis (RA) and drug-induced liver injury (DILI), with its dynamic accumulation closely associated with disease progression. To effectively monitor the level of ONOO- in biological system, fluorescent probes offer distinct advantages for ONOO-, including high spatiotemporal resolution, non-invasive imaging capability, and exceptional sensitivity for detecting transient ONOO- fluctuations in living systems. These probes enable real-time visualization of ONOO- in inflamed joints or drug-stressed livers, facilitating early diagnosis and therapeutic monitoring. Thus, a novel mitochondria-targeted ratiometric near-infrared (NIR) fluorescent probe, DXD, was designed and synthesized via the condensation reaction of a diphenylamino-substituted xanthene with dicyanoisophorone. The incorporation of a dicyanoisophorone bridging unit enabled emission redshift into the NIR region, while a phenylboronic acid recognition group conferred rapid and selective responsiveness to ONOO-. DXD exhibited a markedly larger Stokes shift and faster response kinetics for ONOO-. Importantly, the original emission peak of DXD at 734 nm gradually decreased, while a new peak at 618 nm progressively emerged and intensified, enabling ratiometric detection of ONOO-. Functionally, DXD enabled selective monitoring of dynamic fluctuations in both endogenous and exogenous ONOO- in live cells. It is noteworthy that ONOO- concentrations were significantly elevated in RA and DILI models compared to normal mice, providing important imaging evidence for the role of ONOO- in disease progression. Detecting ONOO- levels using fluorescent probes is critically important because ONOO- acts as a key pathogenic mediator in both RA and DILI models, driving inflammation, autoimmunity, and tissue damage. Furthermore, its real-time monitoring serves as an early and sensitive biomarker for disease progression, often showing changes before traditional clinical indicators.
A sensitive, simple, and environmentally friendly electrochemical voltammetric approach has been developed for the determination of Mirabegron (MRG) in drug substances, drug products, and spiked human plasma samples. For the first time, MRG was measured using a combination of electrochemistry and nanotechnology. A Multiwalled carbon nanotubes/sugar polymer/ zinc oxide nanoparticles carbon paste electrode (MWCNT/ZnO NPs/PS/CPE) was fabricated and used to study the electrochemical behavior of MRG showing a good anodic response in Britton-Robinson buffer (B-R buffer) pH 3.0 utilizing cyclic (CV) and square wave voltammetry (SWV). Linear relationships were recorded between the peak current (Ip ) and MRG concentration ranges of 0.2 × 10- 9 - 100 × 10- 6 M in bulk with correlation 0.9990 and 0.30 × 10- 9 - 100 × 10- 9 M in plasma with correlation of 0.9993. Method sensitivity was demonstrated by the calculation of the detection and quantification limits, which were found to be 6 × 10- 12 M and 19 × 10- 12 M, respectively. Method validation has been evaluated as per ICH guidelines. The acquired results were statistically compared with those of the reported one profitably. Method greenness was evaluated by ECO scale, the Modified Green Analytical procedure index (MOGAPI) and Analytical Greenness Metric Approach (AGREE). The results indicate an excellent green profile, so the developed new voltametric method will be dedicated to the quality control laboratories to contribute to MRG analysis.
The pharmaceutical industry's shift toward new drug modalities, including therapeutic peptides, modified oligonucleotides, and antibody-drug conjugates, has exposed fundamental gaps in cheminformatics infrastructure. Unlike small molecules, which benefit from mature representation standards and reliable data exchange, new modalities lack robust and interoperable systems capable of capturing their structural and chemical complexity. Drawing on our experience at AstraZeneca, we examine these challenges across peptides, oligonucleotides, and ADCs, focusing on the limitations of current approaches, particularly HELM. We show that these limitations arise from both technical constraints, as new modalities exceed the scope of purely atomistic or sequence-based representations, and organizational gaps, including unresolved standardization and governance. We argue that local solutions exacerbate fragmentation, and that vendor- and community-driven standards, open implementations, and stronger governance are required to enable standardized and interoperable chemical information systems for next-generation therapeutics.
Dysmenorrhea is a common gynaecological condition among young women, with non-steroidal anti-inflammatory drugs (NSAIDs) recommended as first-line therapy. However, a substantial proportion of women experience inadequate pain relief despite NSAID use. This study aimed to determine the prevalence of NSAID-resistant dysmenorrhea and to identify predictors as well as alternative pain management strategies among female undergraduate students at Afe Babalola University, Ado-Ekiti, Nigeria (ABUAD). A descriptive cross-sectional study was conducted among 271 female undergraduate students of ABUAD. Data were collected using a structured, self-administered electronic questionnaire that assessed socio-demographic characteristics, menstrual history, dysmenorrhea severity, NSAID use, perceived effectiveness, and alternative pain management strategies. NSAID-resistant dysmenorrhea was operationally defined as persistent menstrual pain despite reported use of recommended NSAID dosages. Descriptive statistics were used to summarize data. Bivariate analyses were conducted using Chi-square, Fisher's exact, and Mann-Whitney U tests, as appropriate. Multivariate logistic regression was performed to identify independent predictors of NSAID resistance. Statistical significance was set at p ≤ 0.05. The prevalence of dysmenorrhea among participants was 70.8% (192/271). Of those with dysmenorrhea, 62.5% (120/192) reported NSAID use for pain management. Among NSAID users, 63 students were classified as having NSAID-resistant dysmenorrhea, representing 52.5% of NSAID users and 32.8% of all students with dysmenorrhea. Perceived delayed onset of NSAID action (adjusted odds ratio [AOR] = 16.91; 95% CI: 2.00-141.60; p = 0.009) and lower NSAID effectiveness scores (AOR = 0.72; 95% CI: 0.57-0.90; p = 0.004) were significant predictors of NSAID resistance. Common coping strategies among NSAID-resistant students included activities such as use of alternative medications (25.4%), NSAID dose escalation (23.8%), rest or sleep (6.3%), as well as medications including antispasmodics (31.7%), and herbal remedies (11.2%). NSAID-resistant dysmenorrhea is common among undergraduate students. Perceived delayed onset and reduced effectiveness of NSAIDs are key predictors of resistance and contribute to reliance on alternative and potentially unsafe coping strategies. These findings highlight the need for improved education on appropriate dysmenorrhea management, early identification of NSAID non-responders, and access to evidence-based alternative treatment options. Painful menstruation (dysmenorrhea) is common among young women and can affect daily activities and academic performance. Non-steroidal anti-inflammatory drugs (NSAIDs), such as ibuprofen and diclofenac, are commonly used to treat menstrual pain. However, some women do not experience adequate relief even when these medicines are taken correctly.This study investigated how often menstrual pain does not respond to NSAIDs, the predictors of poor response, and the alternative ways students manage their pain. The study included 271 female undergraduate students at Afe Babalola University in Nigeria.Most participants reported experiencing menstrual pain, and many used NSAIDs for relief. However, more than half of NSAID users reported that the medicines did not adequately relieve their pain, even when taken at the recommended doses. Overall, nearly one in three students with menstrual pain was classified as having NSAID-resistant dysmenorrhea. Many students reported that NSAIDs worked too slowly or were ineffective. When NSAIDs did not relieve pain, students often used other strategies such as increasing NSAID doses without medical advice, taking other pain medicines, resting, using heat therapy, herbal remedies, or visiting the hospital. Students who perceived NSAIDs as slow or ineffective were more likely to have NSAID-resistant menstrual pain, while factors such as age and menstrual characteristics were not strongly associated.These findings highlight the need for better education on safe pain management, early identification of students who do not respond to NSAIDs, and improved access to effective treatment options for menstrual pain.
Exosomes, which transport miRNAs in vivo, hold significant therapeutic potential for treating diseases. Current methods for loading miRNAs into exosomes include sonication, co-incubation, kit-based transfection, and electroporation, with electroporation being the most efficient and widely used approach. However, standardized protocols for electroporation conditions remain lacking, necessitating the optimization of electroporation parameters to enhance the utility of extracellular vesicles as drug delivery vehicles in vivo. Platelets were isolated from healthy volunteer blood donors, and platelet-derived exosomes were extracted. The exosomes were labeled with specific dyes and loaded with miRNA using Bio-Rad Gene Pulser Electroporation buffer or 50 mM trehalose. Electroporation was performed at 150 V, 350 V, and 500 V. The miRNA-loaded exosomes were then co-incubated with cells. The efficiency of miRNA delivery was evaluated through fluorescence co-localization, nanoparticle tracking analysis, and qPCR. Our findings demonstrate that the Bio-Rad Gene Pulser Electroporation buffer is highly effective as an electroporation medium for exosomes. Optimal miRNA transfection efficiency and cellular uptake were achieved at 350 V, with significantly higher exosome internalization observed under these conditions. Utilizing the Bio-Rad Gene Pulser Electroporation buffer at 350 V enhances both miRNA loading efficiency into extracellular vesicles and subsequent cellular uptake. This study establishes an optimized electroporation protocol, addressing limitations in existing methodologies and advancing the potential of extracellular vesicles as a robust platform for miRNA-based therapeutic delivery.
Trypanosoma cruzi is a digenetic parasite that undergoes various transformations to complete its life cycle. Changes between hosts and vectors involve exposure to stressful environments, for which it has developed different strategies to cope with such stress. L-threonine 3-dehydrogenase (TDH) is a key enzyme in trypanosome metabolism. A recent study has shown that inhibiting TDH reduces glycine and acetate production, which disrupts parasite growth and viability. Furthermore, altering the threonine catabolism pathway has been suggested to impact redox homeostasis by changing the NAD+/NADH ratio and the production of downstream metabolites. To understand the role of TDH in T. cruzi, we overexpressed this gene and assessed some biological parameters after exposure to reactive oxygen species (ROS), alkylating agents, and the drug benznidazole (Bz), which causes oxidative stress and genetic damage. Our results show that the TDH enzyme led to higher survival rates under H2O2 exposure and increased tolerance to Bz. Moreover, the parasites showed a higher infection rate and preserved mitochondrial membrane potential stability, both of which are associated with greater tolerance to ROS. Finally, parasites overexpressing TDH were less vulnerable to genetic damage caused by agents such as methyl methanesulfonate (MMS) and gamma radiation. Overall, our results demonstrate that TDH, a key enzyme in threonine metabolism, helps combat stressful environments and supports parasite survival.
Antibody-drug conjugates (ADCs) are rapidly evolving from conventional cytotoxic delivery systems into multifunctional, immune-integrated therapeutic platforms. Highlights from the 2026 American Association for Cancer Research (AACR) Annual Meeting demonstrate significant advances in ADC design, including dual- and multi-payload constructs, multispecific targeting strategies, and immunostimulatory payloads. These innovations aim to overcome key limitations such as tumor heterogeneity, resistance, and systemic toxicity. Novel approaches targeting the tumor microenvironment, including depletion of regulatory T cells and tumor-associated macrophages, further expand the therapeutic scope of ADCs beyond direct tumor cell killing. In parallel, integration with emerging modalities such as engineered CD16-enhanced natural killer T (NKT) cells underscores the potential for synergy between ADCs and cellular immunotherapies. Advances in AI-guided target discovery and antibody engineering are also enhancing tumor selectivity and internalization. Collectively, these developments highlight a paradigm shift toward precision, multi-mechanistic ADCs with the potential to improve clinical outcomes across diverse cancer types.
An effective stent for ureteral obstruction requires controlled swelling behaviour, sustained drug delivery, inhibition of bacterial adhesion, and performance characteristics that conventional stents frequently fail to attain. Two bi-layer bioresorbable stents (Carr/Alg and Ch/PVA) were developed using carrageenan (Carr) and chitosan (Ch) as the inner layer for drug diffusion and integrated with alginate (Alg) and PVA as the outer layer that provides controlled degradation and structural stabilization. Drugs, including aspirin (Asp) and dexamethasone (Dex), were incorporated facilitating drug delivery. Swelling analysis and degradation study of the stent in artificial urine over 18 days indicated the carrageenan/alginate (Carr/Alg) stent showed high fluid retention of 715.8%, compared to the chitosan/PVA (Ch/PVA) stent with 238.8% maintaining stable shape without excessive swelling. The cumulative drug release for Carr/Alg stent had the highest initial burst of 25% over 48 h, whereas Ch/PVA showed 23%, consistent with non-Fickian model. Surface morphology was analysed, confirming bi-layered structure and porous networks for structural stability. Bacterial adhesion was assessed using gram-negative Escherichia coli, demonstrating anti-adhesion behaviour, while bulging analysis demonstrated dimensional stability, indicating suitability under dynamic urinary tract conditions.
To determine the circumstances of acute poisoning cases at the Poison Control Center (2021-2024) and analyze their risk factors. We retrospectively analyzed the demographic characteristics, clinical features, and prognosis of patients with acute poisoning at the Poison Control Center. This study included 10,402 cases comprising drug poisoning (5,074), alcohol poisoning (2,316), carbon monoxide poisoning (1,803), pesticide poisoning (882), and chemical poisoning (327). The age group with the highest incidence of poisoning was 21-40 years old (37.94%, p < 0.01). Women have a higher proportion of drug poisoning (72.59%, p < 0.01), while men have a higher proportion of alcohol poisoning (74.31%, p < 01). Patients with pesticide poisoning were predominantly educated to the middle school level or below (73.02%, p < 0.01), whereas those with alcohol poisoning were mainly educated to the high school level or above (73.88%, p < 0.01). Carbon monoxide and pesticide poisoning occur primarily in rural areas. Poisoning incidents at home accounted for 74.32%, alcohol poisoning frequently occurred in entertainment venues (60.58%, p < 0.01). Carbon monoxide poisoning is more common in January and December, whereas drug poisoning has a higher incidence throughout the year. Carbon monoxide, chemical and alcohol poisoning were predominantly accidental, whereas drug and pesticide poisoning were mainly intentional. Gastric lavage dominated pestic ide poisoning; Antidotes prevailed in alcohol poisoning.The overall mortality rate was 1.56 %, highest for pesticide poisoning (8.28%, p < 0.01). Different types of poisoning have distinct sociodemographic characteristics, that should be considered when developing prevention and treatment policies. Personalized treatment plans should be tailored to different poisonings.
E3112 is a recombinant human hepatocyte growth factor (HGF) intended for the treatment of acute liver failure. As the presence of anti-drug antibody (ADA) against E3112 could pose a significant risk in clinical settings if it cross-reacts with the body's natural HGF, we have developed assays for E3112 and its ADA in human serum. Assays of E3112 and its ADA developed by ligand binding assays were validated in accordance with bioanalytical guidelines and applied to clinical pharmacokinetic (PK) and immunogenicity assessments. These assays demonstrated the ability to detect E3112 at a concentration as low as 0.156 ng/mL, whereas the sensitivity of ADA was determined to be 42.3 ng/mL. The validation studies, incorporating quality control for the PK assay and positive control of ADA, substantiated the reproducibility of the assays. The ADA and PK assays were applied to the real sample assays supporting a clinical trial of E3112. Following the intravenous administration of E3112, serum E3112 levels declined with a half-life of 19.3 h. No ADA was detected in predose or postdose samples. These findings collectively indicate that E3112 exhibited a favorable PK profile with minimal immunogenicity within the clinical context. E3112 is a new protein-based medicine being developed to treat acute liver injury. To understand how this drug works in the body, it is important to measure both the drug levels in the blood and whether the body produces anti-drug antibodies (ADA), which may affect its safety or efficacy. In this study, we developed simple and reliable assays to measure E3112 and its ADA in human serum. These methods were fully validated according to regulatory guidelines and were successfully used in a clinical study to evaluate how E3112 behaves in the body (pharmacokinetics) and whether it triggers immune responses (immunogenicity).Using these methods, we showed that E3112 can be accurately measured in human serum, and no ADA was detected after administration in the clinical study. These results suggest that E3112 has a favorable pharmacokinetic profile and a low risk of immune reactions in humans. Overall, this study provides practical tools and important information that help clinicians and researchers better understand how E3112 behaves in the body.
Hyaluronidase (HAase) is a clinically important glycosidase that functions both as a drug diffusion enhancer and a disease biomarker. The quantification of HAase activity in biopharmaceutical products has received limited attention and still relies on conventional methods. Furthermore, the rapid and accurate determination of HAase activity is of great significance for cancer diagnosis and could facilitate the development of non-invasive examination methods. These considerations highlight the urgent demand for developing novel, rapid and accurate strategies for detecting HAase activity. In this work, we developed a "signal-on" electrochemical sensor based on an enzyme-responsive hydrogel for the sensitive and rapid detection of HAase activity. The sensor was constructed by crosslinking hyaluronic acid (HA) and polyethylenimine (PEI) to form a hydrogel that electrostatically encapsulates methylene blue (MB). HAase-triggered hydrogel degradation releases MB, generating a measurable current increase. The results demonstrated a strong linear relationship (R2 = 0.9973) between the change in current and HAase activity within the range of 0.78-90 U/mL, with a limit of detection of 0.48 U/mL. The sensor exhibits good specificity, repeatability, and storage stability. Moreover, the sensor has been successfully applied to the analysis of biopharmaceutical products and human urine samples. This facile electrochemical sensor is capable of addressing both drug quality control and disease monitoring. With rapid detection in just 15 min, it significantly improves the efficiency of drug quality evaluation and disease diagnosis. This innovative approach offers a promising tool for the quality evaluation of HAase pharmaceuticals and their diagnostic monitoring.
African animal trypanosomosis poses a significant threat to livestock health and agricultural productivity across sub-Saharan Africa. Isometamidium chloride is the only available drug that is both prophylactic and curative. Despite sustained reports of resistance since the 1970s, a definitive molecular mechanism of resistance remains unresolved in the clinically relevant pathogen species Trypanosoma congolense. In this study, the role of a putative drug/metabolite transporter protein, TcoDMT, was validated via the analysis of in vitro-derived mutants, showing that expression levels of this protein correlated strongly with isometamidium sensitivity. Functional analyses revealed that the protein is a cell surface phenanthridine transporter and, notably, copy number variation correlates with isometamidium sensitivity in T. congolense field isolates. This study validates, for the first time, a plasma membrane transporter with a defined role in phenanthridine action and resistance, advancing our understanding of drug resistance mechanisms in parasitic protists, and informing strategies to combat animal trypanosomosis.
In this study, a novel pH-responsive hybrid nanocarrier with a water-in-oil-in-water (W/O/W) emulsion structure was developed using gelatin (G) as a biocompatible polymer, montmorillonite (MMT) as a layered diffusion barrier, and cerium oxide nanoparticles (CeO₂) as a multifunctional stabilizing agent for pH-responsive and controlled delivery of quercetin (QC). The nanocarriers were synthesized via a double-emulsion method and comprehensively characterized by Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), field-emission scanning electron microscopy (FESEM), and dynamic light scattering (DLS) with zeta potential analysis. The optimized G/MMT/CeO2@QC nanocarriers exhibited a uniform nanoscale size (39.3 nm) and a high negative zeta potential (- 38.6 mV), indicating excellent colloidal stability. Incorporation of MMT and CeO₂ significantly enhanced drug loading and encapsulation efficiency (43.0% and 84.5%, respectively) compared to the MMT-free G/CeO₂@QC system, due to synergistic effects of layered silicate confinement, gelatin-mediated hydrogen bonding, and CeO2-driven Lewis acid-base coordination. In vitro release studies demonstrated pronounced pH sensitivity, with sustained release at physiological pH (60% at pH 7.4 after 96 h) and accelerated release under tumor-mimicking acidic conditions (95% at pH 5.4). To further interpret the release kinetics, machine learning-assisted, shape-constrained data analysis was employed to provide time-resolved and physically consistent insights into pH-dependent release behavior. Kinetic modeling confirmed Higuchi and Korsmeyer-Peppas-controlled diffusion mechanisms. Cytocompatibility and anticancer activity were evaluated using the MTT assay on A549 lung cancer cells and L929 fibroblasts. Blank nanocarriers were non-toxic (> 95% cell viability), while drug-loaded nanocarriers achieved selective cytotoxicity (A549 viability reduced to 55% with 93% viability in L929 cells), outperforming free QC. Overall, this tri-component hybrid system provides a multifunctional nanoscale platform with controlled drug release, high encapsulation efficiency, and tumor-selective cytotoxicity, demonstrating strong potential as a pH-responsive nanocarrier for lung cancer therapy.
Hydrogel-based controlled release systems are widely used in pharmaceutical and agricultural fields. To achieve controlled release of hydrophilic drugs, which otherwise release rapidly through hydrated polymer networks, we developed a facile ferric ion co-crosslinking strategy to fabricate composite hydrogel beads. The highly porous composite hydrogel beads were prepared by blending recycled household formaldehyde adsorbent (RFA) with sodium alginate (Alg) at a 1:1 (w/w) ratio, followed by Fe3+-induced crosslinking and subsequent surface functionalization with tannic acid (TA). Through altering the Alg hydrogel structure, RFA enabled the hydrogel to synergistically restrict drug molecules, and coupled with the blocking function of MPC, this composite structure provided effective regulation of water-soluble drug molecule release. The carrier demonstrated pronounced swelling and disassembly of the metal-polyphenol microfilm, resulting in rapid GV release under alkaline pH conditions. In acidic environments, the conversion of ternary metal-polyphenol complexes into double or single complexes weakened the "on-off" effect, thereby accelerating GV release compared to neutral conditions. The release of GV was governed by combination of diffusion-controlled processes and progressive hydrogel bead relaxation. Additionally, the carrier exhibited no detectable toxicity in murine fibroblast cells and rice seed germination, indicating its favorable biosafety. These findings demonstrated that the controlled-release system had potential to be further explored for environmental protection and hazardous substance management.
Biomolecular interactions involving proteins, nucleic acids, and small molecules constitute the molecular foundation of cellular regulation, signaling, and therapeutic intervention. Advances in mass spectrometry-based proteomics have enabled the systematic characterization of these interactions at unprecedented depth, sensitivity, and structural resolution. This chapter provides a comprehensive overview of state-of-the-art proteomics methodologies developed to investigate protein-protein, protein-nucleic acid, and protein-drug interactions, with particular emphasis on experimental design, sample preparation, and data quality control. Targeted and untargeted strategies are discussed, including affinity purification-mass spectrometry, proximity-dependent labeling, cross-linking mass spectrometry, blue native electrophoresis, and size-exclusion chromatography-mass spectrometry for protein-protein interactions; affinity capture, EMSA-MS, chromatin immunoprecipitation-mass spectrometry, CRISPR-based locus-specific enrichment, and CLIP-based approaches for protein-nucleic acid complexes; and chemoproteomics, thermal proteome profiling, and label-free structural proteomics for protein-drug interaction analysis. The chapter further highlights recent technological innovations, computational tools, and integrative multi-omics strategies that enhance interaction mapping across biological scales. By critically evaluating the strengths, limitations, and appropriate applications of each methodology, this work aims to provide practical guidance for researchers seeking to design robust interactomics experiments and to interpret complex molecular networks in both basic and translational research contexts.