Depression is a chronic mental disorder with high disability and mortality rates, affecting approximately 95 million individuals in China alone. The limitations of current antidepressant therapies, including restricted efficacy, delayed onset, and significant adverse effects, have motivated the search for alternative treatments from traditional Chinese medicine. Psychotria rubra (PR), a widely distributed medicinal plant in southern China with reported antidepressant properties, represents a promising candidate; however, the molecular mechanisms underlying its antidepressant effects remain largely unexplored. This study aims to elucidate the mechanistic basis of PR in depression treatment by integrating network pharmacology, molecular docking, molecular dynamics simulation, and in vivo experimental validation. We integrated two depression-related transcriptome datasets [GSE76826 (training set: 10 depressed vs. 12 healthy blood samples) and GSE98793 (validation set: 64 depressed vs. 64 healthy blood samples)] to identify differentially expressed genes in depression. Ferroptosis-related genes were retrieved from GeneCards (score > 8) and FerrDb databases, and eight machine learning models with five-fold cross-validation were employed to screen key diagnostic genes. Molecular docking and dynamics simulations validated interactions between PR's active ingredients and target genes. In vivo, eighty male C57BL/6 mice (6-week-old) were randomly allocated into eight groups (n = 10 per group): control, chronic unpredictable mild stress (CUMS) model, deacetyl asperulosidic acid (10 mg·kg⁻1·d⁻1, intraperitoneal), deacetyl asperulosidic acid methyl ester (10 mg·kg⁻1·d⁻1, intraperitoneal), Procyanidin B1 (20 mg·kg⁻1·d⁻1, intraperitoneal), Procyanidin B2 (20 mg·kg⁻1·d⁻1, intraperitoneal), Procyanidin C1 (20 mg·kg⁻1·d⁻1, intraperitoneal), and fluoxetine (10 mg·kg⁻1·d⁻1, oral gavage, positive control). Following 21 days of CUMS induction, drug treatment was administered for 14 consecutive days. Behavioral assessments (sucrose preference test, open field test, tail suspension test, and forced swimming test), inflammatory factors (serotonin, interleukin-1β, interleukin-6, tumor necrosis factor-α), oxidative stress markers (superoxide dismutase, malondialdehyde), and ferroptosis indicators (reactive oxygen species, glutathione, Fe2⁺ levels) were measured. Machine learning analysis identified EPAS1, MAPK14, and VEGF-A as candidate diagnostic genes associated with depression and ferroptosis. Molecular docking revealed favorable binding affinities between multiple PR active ingredients (including asperuloside, asperulosidic acid, deacetyl asperulosidic acid, and deacetyl asperulosidic acid methyl ester) and EPAS1, with deacetyl asperulosidic acid showing the most favorable docking score (− 8.737 kcal/mol) and asperuloside and deacetyl asperulosidic acid methyl ester exhibiting comparable binding free energies (− 35.22 and − 35.26 kcal/mol, respectively) in molecular dynamics simulations. In vivo, PR's main active ingredients significantly improved depressive-like behaviors and reduced neuroinflammation, oxidative stress, and ferroptosis-related markers in CUMS-induced mice, with Procyanidin B1 demonstrating the most pronounced antidepressant-like effect. PR alleviated depression potentially through modulating EPAS1 expression. Procyanidin B1 might be the key active ingredients in PR to alleviate the progression of depression and ferroptosis. This study offers a promising direction for developing new antidepressants from traditional Chinese medicine with multi-component, multi-target effects.
Therapy of breast cancer (BC) overexpressing the pro-survival protein Bcl-2 remains a major clinical challenge. The discovery of natural anticancer drugs with potential as Bcl-2 inhibitors has generated increasing interest. The medicinal plant Teucrium polium L. (TPL) exhibits pharmacological properties, including anticancer activities. This study aimed to assess the antiproliferative and pro-apoptotic effects of TPL extracts on BC cells and to predict the potential of TPL extract-derived metabolites as Bcl-2 inhibitors. Plant extraction was performed using several solvents, and cell viability was determined. The expression of proteins and genes related to apoptosis was evaluated using Western blot, the Proteome Profiler™ Human Apoptosis Array Kit, and qPCR, respectively. Caspase-3/7 and mitochondrial permeability transition pore opening (mPTPO) activities were visualized. GC/MS analysis, molecular docking, and in silico modeling were used for metabolite identification, prediction of their molecular interactions with Bcl-2, and their pharmacokinetic profiles. Among extracts, TPL methanolic extract (TPLME) dose-dependently reduced hormone-dependent and triple-negative BC cell viability, while sparing normal mammary epithelial cells. TPLME induced apoptosis by increasing caspase-3/7 activity, activating distinct apoptotic pathways, and modulating BAX, TP53, and BCL-2 gene expression. Differential BCL-2 expression levels in TPLME-treated BC cells were confirmed by variations in mPTPO activity. After TPLME-derived metabolite identification, molecular docking revealed interactions between key metabolites and the active site of Bcl-2. Predictive analysis revealed the safe pharmacokinetic profiles of TPLME-derived metabolites. These findings highlight the promising potential of TPLME-derived metabolites for the development of novel Bcl-2 inhibitors for BC cells, requiring further chemical development and preclinical investigations using in vivo BC models.
Abstract Non-responsive cycles during ovulation induction with letrozole (LZ) often necessitate the use of human menopausal gonadotropin (HMG), which may be associated with undesirable adverse effects. Therefore, strategies to improve the therapeutic outcomes of LZ are needed. This trial aimed to evaluate the impact of combining alpha-lipoic acid (ALA) with LZ on ovulation induction in women with polycystic ovary syndrome (PCOS). This randomized controlled trial included 151 infertile women with PCOS who were randomly allocated to either the LZ group ( n = 76) or the ALA group ( n = 75). The LZ group received LZ (2.5–7.5 mg) with lifestyle modification for up to three treatment cycles or until pregnancy occurred. The ALA group received the same regimen in addition to ALA (1800 mg/day), which was discontinued on the day of human chorionic gonadotropin (HCG) injection in ovulatory patients. Ovulation and pregnancy rates were assessed, along with hormonal parameters. Ovulation per cycle was significantly higher in the ALA group [153/166 (92.2%)] compared with the LZ group [146/203 (71.9%)] ( P = 0.035). Moreover, pregnancy per cycle was significantly higher in the ALA group than in the LZ group ( P = 0.033). Although serum estradiol, mid-luteal progesterone, and endometrial thickness showed numerically higher values in the ALA group, the differences between the groups were not statistically significant after Bonferroni adjustment. The addition of ALA to LZ improved ovulation and pregnancy outcomes and was associated with favorable metabolic and hormonal effects in women with PCOS.
Ensuring equitable access to dispensed-to-patient (DTP) medical devices is an important policy priority for health insurance systems. This mixed-method study examined stakeholder perspectives on barriers and policy priorities for developing a DTP medical-device formulary for health insurance beneficiaries in Saudi Arabia. An exploratory online survey was distributed to 58 purposively selected stakeholders, and 17 responded (29.3
The gut microbiota plays a critical role in regulating systemic immune responses and has emerged as a key determinant of therapeutic efficacy and toxicity in cancer immunotherapy. Accumulating evidence indicates that specific microbial taxa and microbiota-derived metabolites modulate antitumor immunity by shaping immune cell maturation, cytokine signaling, and the tumor microenvironment. In particular, gut microbial metabolites such as short-chain fatty acids, bile acids, and inosine influence immune checkpoint inhibitor responses by regulating T-cell activation, dendritic cell function, and immune homeostasis beyond the intestinal compartment. Preclinical and clinical studies have demonstrated that alterations in gut microbiota composition are associated with variability in immunotherapy outcomes, including treatment resistance and immune-related adverse events. Importantly, microbiota-targeted interventions such as dietary modulation, probiotics, prebiotics, antibiotics, and fecal microbiota transplantation have shown promise in enhancing immunotherapy efficacy and reducing toxicity. This review synthesizes current mechanistic insights and clinical evidence linking the gut microbiota to systemic immunity and cancer immunotherapy outcomes, highlighting microbiome modulation as a potential therapeutic adjuvant to optimize immunotherapy response and support precision oncology.
Type 2 diabetes mellitus (T2DM) is a major contributor to global morbidity, mortality, and healthcare expenditure. In the context of Saudi Arabia, it is concerningly notable that the prevalence of T2DM is particularly high and rising. Existing cost estimates have previously relied rely on aggregate methodologies, potentially an approach that limits their usefulness for hospital-level decision-making. Accordingly, the primary goal in the current study was to estimate the direct medical costs of T2DM using a real-world, bottom-up micro-costing approach. We carried out a retrospective, cross-sectional cost-of-illness study using electronic medical records from a tertiary hospital in Saudi Arabia. The data collection happened between January and December, 2024. We included adult patients with T2DM, and we calculated direct healthcare costs from the hospital payer perspective, including outpatient care, hospitalizations, laboratory investigations, and medications. In data analysis, we used descriptive statistics, one-way ANOVA, and log-transformed ordinary least squares regression in order to assess cost differences and identify predictors of total cost. A total of 431 patients participated and findings showed that the mean annual cost per patient was SAR 15,396.24 (4,103 USD), and a total cost of SAR 6,635,779.57 (1,768,209 USD). Medications (52.9
Cardiovascular diseases (CVDs) continue to be the world's leading cause of death and a major global healthcare burden. Concerns about side effects and long-term therapy restrictions have raised interest in supplementary therapeutic approaches, even if traditional pharmaceutical and surgical procedures are still crucial to the management of CVD. Evidence from published experimental, clinical, and mechanistic investigations that were indexed in PubMed, Scopus, Web of Science, and Google Scholar between 2014 and 2025 is compiled in this review. Through their antioxidant, anti-inflammatory, lipid-lowering, antihypertensive, and endothelial-protective properties, plant-derived phytochemicals such as flavonoids, carotenoids, polyphenols, glucosinolates, and sulfur-containing compounds have considerable cardioprotective potential. Vascular function, oxidative stress, platelet aggregation, insulin sensitivity, and lipid metabolism have all benefited by compounds like quercetin, catechins, curcumin, resveratrol, and lycopene. However, its therapeutic translation is still constrained by issues with bioavailability, dosage uniformity, and clinical validation. Although more extensive clinical research is needed to determine their long-term efficacy and safety, the evidence now available indicates that phytochemicals may be beneficial supplements to traditional cardiovascular treatments.
Osteoporosis is a prevalent metabolic bone disease that requires multi-target therapeutic strategies. Formononetin (FMN), a bioactive isoflavone derived from Astragalus species, has shown potential in promoting bone health; however, its systemic mechanisms remain poorly understood. In this study, we evaluated the anti-osteoporotic effects of FMN using a glucocorticoid-induced bone loss model in zebrafish. Our results demonstrate that FMN significantly and dose-dependently promotes bone mineralization and effectively reverses glucocorticoid-induced skeletal damage. By integrating network pharmacology and molecular docking with experimental validation, we identified a multi-target mechanism underlying FMN's protective effects. FMN was found to regulate several key signaling pathways, including the PI3K-AKT and estrogen pathways, and its therapeutic impact was confirmed by the rectified expression of core genes involved in bone homeostasis and inflammation. This study reveals that FMN exerts its anti-osteoporotic effects through the coordinated regulation of growth factor signaling, calcium homeostasis, and the oxidative stress-bone metabolism axis. Our findings provide a systematic theoretical foundation for developing FMN as a promising natural therapeutic agent for osteoporosis through the modulation of osteoimmunology.
The N-methyl-D-aspartate receptors (NMDARs) are ionotropic glutamate receptors in the central nervous system (CNS) that play a crucial role in synaptic plasticity, learning, and memory. However, NMDAR overstimulation is a critical pathological feature in neurodegenerative diseases, stroke, epilepsy, and traumatic brain injury. This excitotoxic cascade triggers massive intracellular calcium influx, oxidative stress, and mitochondrial dysfunction, ultimately driving neuronal apoptosis. In this regard, phytochemicals have emerged as safe, multitargeting NMDAR inhibitors with significant therapeutic potential. This review synthesizes current experimental evidence regarding the neuroprotective potential of major phytochemical classes, including glycosides (e.g., vitexin, ginsenosides), polyphenols (e.g., quercetin, resveratrol), alkaloids (e.g., cytisine, huperzine A), and terpenoids (e.g., bilobalide, fucoxanthin). The results of these studies demonstrate that these bioactive compounds mitigate excitotoxicity via multiple mechanisms, including direct receptor blockade, inhibiting specific subunits (such as GluN2B), and downregulating NMDAR expression. Furthermore, they restore neuronal homeostasis by modulating downstream signaling pathways; thereby suppressing neuroinflammation and apoptosis. It can be concluded that phytochemicals represent a vital reservoir of modulatory agents that protect against glutamate-induced neurotoxicity. Hence, they offer a compelling framework for future drug discovery in CNS therapeutics.
Abstract Background The clinical trial ecosystem of Saudi Arabia has advanced significantly, with innovative industry-sponsored clinical trials (iCTs) growing by 27% from 2018 to 2023, reaching 51.4% in the first half of 2025. However, the Kingdom remains underrepresented in global research. This study aimed to quantify this gap and develop strategies to enhance Saudi Arabia's competitiveness. Methods A mixed-methods research design was employed. Phase 1 involved a quantitative landscape analysis of historical and current iCT data (2016–2023) using the LongTaal Clinical Trial Informatics platform. Phase 2 involved a qualitative assessment of expert input derived from two national clinical trial conferences (2022 and 2024), in which expert panel discussions and documented outputs were used to contextualize findings and inform strategic recommendations. Results In 2023, Saudi Arabia’s iCT market share was < 0.06%, significantly lower than its ~ 0.9% share of global pharmaceutical consumption. This 16-fold disparity highlights a critical lack of patient representation. Economic modelling indicates that achieving parity could increase annual research and development (R&D) investment from ~ USD 46 million to over USD 740 million. Insights derived from conference-based expert discussions highlighted regulatory timelines, infrastructure fragmentation, and insufficient incentives as primary barriers. Conclusion To bridge this gap, the authors propose operationalizing a centralised national body to serve as a "one-stop shop" for global sponsors. Strategic priorities include harmonizing regulatory timelines to 6 months, establishing distributed Centres of Excellence, and implementing direct R&D financial incentives. These measures are essential to ensuring Saudi patients are adequately represented in the development of novel pharmaceuticals.
Orthopedic implant-associated infections (IAIs), predominantly caused by Pseudomonas aeruginosa, represent a major clinical challenge due to antibiotic resistance and biofilm formation, which together contribute to their chronic nature. This research presents a biogenic biofilm-resistant chitosan encapsulated Cu/ZnO nanocoating (CS-Cu/ZnO NC) synthesized using Euphorbia thymofolia metabolites and applied on orthopedic implant surface (Tubular bone locked stainless steel plates). The synthesized CS-Cu/ZnO NCs were characterized physicochemically (UV–Vis, FTIR, XRD, SEM) and biologically (antibacterial, antibiofilm, antivirulence, hemolysis and cytotoxicity). Microstructural analysis using UV–Visible Spectroscopy (UV–Vis), X-ray diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR) and Scanning Electron Microscopy (SEM) confirmed the successful synthesis, crystallinity and integration of Cu/ZnO into the chitosan matrix. SEM confirmed that a thin uniform coating was formed by the layer-by-layer method compared to the conventional dip coating method. CS-Cu/ZnO NCs at MIC level (4 µg/mL) reduced virulence traits of MDR P. aeruginosa, such as motility behavior, pyocyanin production, and biofilm production by 90
Despite multiple therapeutic strategies, a significant proportion of psoriatic patients fail to achieve complete/sustained disease clearance, highlighting the need for effective adjuvant or alternative therapy. Melittin, the main component in bee venom, exhibits valuable anti-inflammatory and immunomodulatory properties in various diseases; meanwhile, its effect on psoriasis has not been explored yet. Our study aims to investigate the anti-psoriatic effect of melittin with possible involvement of phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) and the autophagy pathways. Mice were divided into 5 groups: Control group, Imiquimod (IMQ) group, and groups (3–5) received daily intraperitoneal injections of methotrexate (MTX, 1 mg/kg), melittin (40 mg/kg), or melittin (80 mg/kg), respectively. All groups except the control received topical IMQ for seven days. Melittin 80 demonstrated superior efficacy, significantly reducing the clinical Psoriasis Area and Severity Index score by 60
Among various biodegradable polymers, polylactic-co-glycolic acid (PLGA) is one of most widely studied synthetic polymer for controlled and targeted drug delivery. PLGA is a smart polymer as it has stimuli-responsive behaviour. PLGA is approved by the US FDA for multiple therapeutic applications due to its biodegradability, biocompatibility and sustained-release behaviour. PLGA is commercially available as different molecular weights and copolymer ratios, which allow for tuning polymer behaviour to suit numerous applications. Different methods have been used to process PLGA into diverse morphologies, structures and sizes. Traditional PLGA processing methods face many limitations related to scalability and residual solvents. This review was limited to the emerging hot melt extrusion (HME), a more specific, solvent-free and highly scalable technology for fabricating PLGA-based formulations. Key operational parameters such as glass transition temperature, melt viscosity, and degradation behaviour are discussed alongside with the drug-related stability, dispersion and final solid-state properties that need several factors to be fine-tuned to offer a performance with optimal efficiency. This tunability of characteristics makes PLGA driven by HME ideal for fabrication with advanced manufacturing technologies to deliver surprisingly novel drug delivery platforms that can boost therapeutic efficacy and patient compliance across a range of clinical fields.
Pulmonary arterial hypertension (PAH) is a refractory cardiopulmonary disorder with a high mortality rate and few treatment options. Fisetin, a natural flavonoid, exhibits pleiotropic effects including anti-angiogenic, anti-proliferative, anti-inflammatory, and autophagy-modulating activities. To evaluate the therapeutic potential of Fisetin, we established two experimental models: a rat model of monocrotaline (MCT)-induced pulmonary arterial hypertension (PAH) and a platelet-derived growth factor (PDGF)-induced human pulmonary artery smooth muscle cell (PASMC) proliferation model. The therapeutic efficacy and underlying mechanisms of Fisetin were comprehensively assessed using a combination of in vivo and in vitro approaches, including right heart catheterization, masson’s trichrome and hematoxylin–eosin staining, immunohistochemistry, westernblot, immunofluorescence, ELISA, and transmission electron microscopy. Additionally, Protein–protein docking predicts interactions between proteins, whereas molecular docking predicts interactions between small molecules and proteins. Fisetin significantly ameliorated pulmonary vascular remodeling and reduced both right ventricular systolic pressure and the right ventricular hypertrophy index in monocrotaline (MCT)-induced PAH rats. The underlying mechanism may involve suppression of the TGF-β1/Smad2/3 pathway, which regulates autophagy, cell proliferation, migration, and inflammatory responses. In vitro, Fisetin exerted a dose-dependent suppressive effect on these molecular events. This study suggests that Fisetin may alleviate MCT-induced PAH rats by suppressing aberrant TGF-β1/Smad2/3 pathway activation. These findings provide experimental and theoretical support for Fisetin as a promising therapeutic candidate for PAH.
Ramucirumab, a monoclonal antibody that targets vascular endothelial growth factor receptor-2 (VEGFR-2), is primarily known for its ability to inhibit angiogenesis in a variety of cancer types. Its direct cytotoxic mechanisms in colorectal cancer cells, especially those related to mitochondrial dysfunction and reactive oxygen species (ROS)-mediated apoptosis, are inadequately elucidated. This work aims to examine the involvement of mitochondrial dysfunction, oxidative stress, and caspase activation in mediating ramucirumab induced cytotoxicity in Caco-2 cells. MTT (3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) and lactate dehydrogenase (LDH) tests were used to measure cell viability and cytotoxicity, respectively by exposing Caco-2 cells to ramucirumab for 24 h. Flow cytometry was used to analyze apoptosis and cell cycle distribution. Hoechst 33342, rhodamine 123 and carboxy-H2DCFDA staining was used to examine nuclear condensation, mitochondrial membrane potential (ΔΨm) and intracellular ROS, respectively. Additionally, the expression of the apoptotic markers caspase-3 and caspase-7, and levels of VEGF was quantified. Ramucirumab enhanced the release of LDH, and significantly (p < 0.05) decreased cell viability with increasing concentration and IC50 value was estimated as 780 µg/ ml. A significant (p < 0.05) increase in the apoptotic cell population and cell cycle arrest primarily at the S-phase was observed. Nuclear condensation and fragmentation were observed and mitochondrial membrane potential was found to be decreased, indicating mitochondrial dysfunction. Additionally, oxidative stress (ROS) and VEGF was elevated and cleaved caspase-3 and caspase-7 were significantly upregulated, indicating caspase-mediated apoptosis. In conclusion, by causing cell cycle arrest and inducing apoptosis via mitochondrial malfunction and caspase pathway activation, ramucirumab demonstrates anti-cancer action against Caco-2 cells. These findings provide new perspectives on ramucirumab's wider anti-tumor potential beyond angiogenesis inhibition.
This cross-sectional study examines the use of complementary and alternative medicine (CAM) among patients with multiple sclerosis (MS) in Riyadh. A survey was conducted among 151 multiple sclerosis patients enrolled at the neurology clinics of two hospitals in Riyadh (KSUMC and KFMC). The study indicated that a significant majority of respondents (60.93
In the community pharmacy sector the concept of service quality is important. Pharmacists working in these settings provide useful services that go beyond merely dispensing medications. This study aimed to translate the short Perceived Service Quality Scale (PSQS−SF) to Arabic and to use this scale to assess the determinants of service quality of community pharmacies in relation to competitively priced high quality service and loyalty intention. A pre−validated short Perceived Service Quality Scale (pSQS−A−SF) was translated from English to Arabic, validated and approved by the original authors. The scale consisted of six questions assessing pharmacy service quality, two questions evaluating the perception of price competitiveness, and three questions measuring loyalty intentions. All of the items were rated on a 5−point Likert scale. Community pharmacy users have been approached to answer the survey and descriptive statistics were reported. The pSQS−SF was successfully translated and culturally adapted to Arabic context (pSQS−A−SF) with minor modifications. A total of 207 respondents completed the survey. Perceived service quality was rated highly across most items, whereas perceptions of price competitiveness were lower. In confirmatory factor analysis, the measurement model showed improved fit after minor modification (deletion of one service−quality item), with acceptable factor loadings and composite reliability, although average variance extracted was marginal. Service quality was strongly correlated with loyalty intentions. In the structural model, higher perceived service quality predicted greater patronage loyalty (β=0.26, p=0.023) and prescription loyalty (β=0.41, p=0.002), while price competitiveness did not significantly predict loyalty outcomes; private health insurance showed a small positive association with patronage loyalty (β=0.156, p=0.022). The Arabic version of the perceived service quality short-form demonstrates preliminary evidence of construct validity and reliability and can support evaluation of patient experience in Saudi community pharmacies. Perceived service quality—rather than price competitiveness—was linked with self-reported patronage loyalty in this sample. Further research should strengthen validation (including discriminant validity and broader sampling) and assess performance across settings and subgroups.
Melanoma is a highly heterogeneous malignancy in which tumor behavior is shaped by complex signaling, trafficking, and metabolic programs. Two-pore channel 2 (TPC2), an endolysosomal ion channel, has been implicated in pigmentation, vesicular trafficking, and cancer-related cellular processes, but its role in melanoma remains incompletely understood. In this study, we investigated the impact of TPC2 loss in two melanoma models, the human cell line CHL-1 and the murine cell line B16, using previously validated CRISPR/Cas9-generated TPC2 knockout cells. Proteomic profiling, pathway enrichment analysis, extracellular flux measurements, and mitochondrial membrane potential assessment were used to define the molecular and functional consequences of TPC2 deletion. Proteomic analysis revealed marked but distinct changes in protein expression in CHL-1 and B16 cells, with clear separation between wild-type and TPC2 KO proteomic profiles in both models. Functional enrichment analysis showed that CHL-1 cells were preferentially associated with pathways related to metabolic regulation, morphogenesis, intracellular transport, extracellular matrix organization, and signaling networks including WNT and TGF-β, whereas B16 cells were enriched in immune/interferon-related pathways, protein homeostasis, intracellular trafficking, and stress-response programs. Bioenergetic profiling demonstrated that TPC2 KO altered cellular metabolism in a cell line-dependent manner. In CHL-1 cells, TPC2 loss reduced basal oxygen consumption rate (OCR) and the OCR/ extracellular acidification rate (ECAR) ratio, but enhanced oxidative adaptation under glucose-free conditions. In contrast, B16 TPC2 KO cells displayed increased OCR and ECAR under basal conditions, consistent with a more energetically active phenotype. Despite these differences in basal bioenergetics, TPC2 KO preserved mitochondrial membrane potential under stress in both cell lines, indicating enhanced mitochondrial stress resilience. Collectively, these findings identify TPC2 as a context-dependent regulator of melanoma cell physiology that influences proteomic remodeling, metabolic adaptation, and mitochondrial function. These results support further investigation of TPC2 as a potential therapeutic target in melanoma, while emphasizing that its biological effects depend on cellular context.
Doxazosin mesylate is a selective α1-adrenergic receptor antagonist with low oral bioavailability due to poor aqueous solubility and extensive first-pass hepatic metabolism. This study aimed to design and optimize a doxazosin mucoadhesive buccal film to overcome hepatic metabolism and enable rapid drug onset. The buccal films were prepared by solvent casting, using a Plackett–Burman design to screen formulation variables affecting ex vivo mucoadhesive force, followed by a 22 full factorial design to optimize in vitro drug release by varying the concentrations of HPMC, the main film-forming polymer, and glycerin, a plasticizer. The prepared films were characterized for physico-mechanical properties, surface pH, mucoadhesive force, swelling properties, and drug release. In addition, the optimized formula was further characterized by scanning electron microscopy and ex vivo permeation study. The screening study showed that mucoadhesive strength ranged from 28 to 41 g, with the polymer type, polymer concentration, plasticizer type, and plasticizer concentration identified as the most influential variables. The optimized formula (FF3), containing 66 w/w
Botulism is a severe neuroparalytic syndrome caused by botulinum neurotoxins (BoNTs)—among the most potent biological agents—with an estimated human lethal dose (LD50) of approximately 1 ng/kg. By cleaving soluble N-ethylmaleimide–sensitive factor attachment protein receptor (SNARE) proteins at cholinergic nerve terminals, BoNTs block acetylcholine release, producing potentially fatal flaccid paralysis. Despite advances in supportive care, current therapeutic options remain limited to antitoxin administration, which is effective only against circulating toxin and cannot neutralize intracellular BoNT. Consequently, patients often require prolonged mechanical ventilation and rehabilitation. To address these limitations, this review provides a comprehensive overview of botulism, spanning historical recognition, structural and mechanistic insights into BoNT activity, clinical manifestations, and current treatment strategies, while highlighting therapeutic gaps. Particular emphasis is placed on emerging preclinical interventions, including small-molecule inhibitors, antibody-based therapeutics, intracellular clearance strategies, and gene- and RNA-based modalities, reflecting rapid progress in structural biology and pharmacology. Collectively, these advances highlight both the promise and the remaining translational challenges of developing next-generation countermeasures, with implications for clinical management and biodefense preparedness. Botulism: Pathophysiology, Clinical Presentation, and Therapeutic Landscape. Schematic overview of botulism from exposure to treatment. BoNT exposure may occur through foodborne, infant, wound, iatrogenic, or bioterror-related routes. After systemic distribution, the toxin binds peripheral cholinergic nerve terminals, undergoes endocytosis, and translocates its light chain into the cytosol, where SNARE cleavage blocks acetylcholine release. This results in the characteristic descending paralysis of botulism, with early cranial nerve involvement followed by limb weakness and respiratory failure. The figure also highlights the limited window for antitoxin efficacy before neuronal internalization and summarizes both current therapies and emerging preclinical countermeasures aimed at neutralizing circulating toxin, enhancing intracellular clearance, or restoring neuronal function. ACh, acetylcholine; BIG-IV, botulism immune globulin intravenous; BoNT, botulinum neurotoxin; HBAT, heptavalent botulism antitoxin; IV, intravenous; PROTAC, proteolysis-targeting chimera; RNA, ribonucleic acid; SNAP-25, synaptosomal-associated protein 25; SNARE, soluble N-ethylmaleimide–sensitive factor attachment protein receptor; VAMP, vesicle-associated membrane protein (synaptobrevin)
