RNA extraction from some plants is difficult owing to the presence of pigments and secondary metabolites in vacuoles and cell walls, which hinder molecular biological studies. In this study, we developed a simple method for high-yield RNA extraction from pitchers of the carnivorous plant Nepenthes × ventrata that is recalcitrant to RNA isolation. RNA was extracted from pitchers of Nepenthes × ventrata at different developmental stages using conventional methods. Additionally, a modified cetyltrimethylammonium bromide (CTAB) method, in which an ethanol-precipitation step at high LiCl concentration was introduced, was used to extract RNA. Conventional purification methods yielded low or negligible amounts of RNA from pitcher tissues of Nepenthes, and the extracts contained visible pigment contamination. The conventional CTAB method, which is effective for RNA extraction from sweet potato tissues, also yielded only a small amount of RNA from Nepenthes tissues; most of the RNA remained in the supernatant discarded during the LiCl precipitation step, along with the contaminating pigments. By replacing this step with ethanol precipitation in the presence of high concentration of LiCl, the contaminating pigments were removed, and RNA with high purity and yield was recovered. Purified RNA could be efficiently used for full-length cDNA synthesis. The modified method requires no expensive extraction kits and is relatively safe because it does not use phenol. Therefore, this method may be effectively applied to plants that are recalcitrant to RNA isolation.
Açaí (Euterpe oleracea) is widely recognized as a functional food due to its nutritional value and abundance of bioactive compounds. Its seeds, representing approximately 80%-95% of the fruit mass, constitute an abundant agro-industrial byproduct with promising biotechnological potential. This study aimed to chemically characterize a 70% ethanolic extract obtained from E. oleracea seeds through liquid chromatography-mass spectrometry (LC-MS) annotation and total phenolic content (TPC) determination, as well as to investigate its antioxidant activity, prebiotic potential, cytotoxicity, and mutagenic safety profile. The extract was obtained by percolation using 70% ethanol. Antioxidant activity was evaluated by the DPPH assay, while prebiotic potential was assessed through submerged in vitro fermentation with Lactiplantibacillus plantarum LP90, monitoring bacterial growth and medium acidification. Cytotoxicity was evaluated in Caco-2 cells using a resazurin assay, and mutagenicity was investigated using the micronucleus and Ames tests. LC-MS annotation revealed a phenolic-rich profile, mainly composed of procyanidins. The extract showed a TPC of 123.12 ± 5.70 mg GAE/g dry weight and significant antioxidant activity (EC50 = 11.79 ± 0.9 µg/mL). Additionally, the extract promoted L. plantarum growth and reduced medium pH, suggesting fermentable substrate potential. The extract also demonstrated low cytotoxicity (IC50 = 704.4 ± 41.8 µg/mL) and no mutagenic effects. These findings support the valorization of E. oleracea seeds as a potential source of bioactive compounds for functional and nutraceutical applications.
Toxic hepatitis is characterized by enhanced oxidative stress and disruption of antioxidant defense mechanisms in the liver mitochondria. In this study, we investigated the effects of polyphenolic extracts (Helmar-1 and Helmar-2) obtained from Helichrysum maracandicum on mitochondrial antioxidant systems in rats with experimentally induced toxic hepatitis. Experimental toxic hepatitis was established in rats through intraperitoneal administration of carbon tetrachloride (CCl₄) diluted in olive oil (50%, 1 mL/kg), administered twice weekly over a two-week period. Liver injury was verified by elevated plasma levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST). The animals were subsequently treated with Helmar-1 and Helmar-2 extracts at a dose of 20 mg/kg per day for 10 consecutive days. The activities of key antioxidant enzymes (superoxide dismutase, catalase, and glutathione peroxidase), malondialdehyde (MDA) content, and mitochondrial respiration and oxidative phosphorylation parameters (V₂, V₃, V₄, RCR, and ADP/O ratio) were assessed in liver mitochondria. Treatment with both Helmar-1 and Helmar-2 extracts resulted in a significant enhancement of antioxidant enzyme activities, a marked reduction in MDA levels, and substantial improvement in mitochondrial respiration and oxidative phosphorylation parameters compared to untreated toxic hepatitis groups. Overall, these results confirm that Helichrysum maracandicum polyphenol extracts improve mitochondrial respiration, oxidative phosphorylation efficiency, antioxidant defense, and membrane stability in toxic hepatitis.
The Kelch-like ECH-associated protein 1 (Keap1)-Nuclear factor E2-related factor 2 (Nrf2) pathway is integral to cellular defense mechanisms against oxidative and xenobiotic stress by neutralizing reactive oxygen species (ROS). An increase in ROS levels is a significant factor in the advancement of neurodegenerative diseases (NDDs). Consequently, the development of effective Keap1 inhibitors that disrupt the Keap1-Nrf2 interactions and subsequently enhance Nrf2's transcriptional activity may represent promising therapeutic avenues for NDDs. Given that Nardostachys jatamansi (N. jatamansi), a medicinal plant, has shown potential as a treatment for Parkinson's disease (PD), we sought to assess its influence on the Keap1-Nrf2 pathway. Our research demonstrates that the phytochemicals in the methanol extract of N. jatamansi can activate the Keap1-Nrf2-ARE signalling cascade. Computational analyses, including molecular docking and molecular dynamics simulations with AutoDock Vina and Desmond, respectively, revealed the binding properties of selected N. jatamansi compounds, of which 16 phytocompounds were confirmed in the methanol extract using LC/MS. In vitro assays demonstrated a dose-dependent interaction of the extract with both the BTB and β-propeller domains of Keap1, as well as inhibition of the BTB-Cullin3 interaction. The extract was assessed for its ability to protect N27 dopaminergic neurons from rotenone-induced neurotoxicity by activating the Nrf2-ARE pathway. It demonstrated dose-dependent ARE transactivation, suggesting Nrf2 dissociation from Keap1. Altogether, these observations indicate that phytocompounds derived from N. jatamansi offer significant potential for therapeutic or neuroprotective applications in Parkinson's disease.
Lichen planus is a chronic autoimmune inflammatory mucocutaneous disease affecting the skin and oral mucosa. The objectives of the study were to evaluate and compare the effectiveness of topical Amlexanox, Curcuma longa extract, and clobetasol propionate in the treatment of oral lichen planus (OLP). A total of 45 patients with OLP were randomly divided into three groups: Group I (Amlexanox), Group II (Curcuma longa extract), and Group III (Clobetasol propionate). Baseline recordings of pain and burning sensation were conducted using the visual analog scale (VAS) and numeric rating scale (NRS). Clinical signs of erythema, ulceration, and lesion size were evaluated and recorded. All patients were evaluated at weekly intervals during the 2-month therapy period, as well as during follow-up for 3 months (at 15-day intervals). Amlexanox showed a significant reduction in pain and burning sensation, along with a decrease in clinical signs of OLP. However, complete remission was not achieved, and mild reappearance of erythema was observed. Curcuma longa extract also showed a significant reduction in pain and burning sensation, with a considerable reduction in clinical signs to a tolerable level. This reduction remained static during the follow-up period. Clobetasol propionate demonstrated higher efficacy in reducing pain, burning sensation, and clinical signs of OLP. Although complete remission was achieved during the therapy period, the reappearance of pain, burning sensation, and clinical signs was noted during follow-up visits. Amlexanox and Curcuma longa extract reduce the signs and symptoms of OLP to a tolerable level, with improvement remaining static after the cessation of therapy. Clobetasol propionate causes complete remission of signs and symptoms of OLP, but there is a higher chance of recurrence.
5-Hydroxymethylfurfural (HMF) is a biomass-derived platform chemical with significant potential in the sustainable production of fuels and value-added chemicals. However, its large-scale synthesis from glucose in aqueous media is limited by humin formation arising from HMF accumulation, as well as its instability under acidic reaction conditions. These challenges highlight the need for rapid in situ extraction strategies to stabilize HMF and suppress side reactions. Accordingly, the development of environmentally benign and economically viable catalytic systems that enable efficient HMF extraction while maintaining catalytic performance is critical. Herein, molecular dynamics (MD) and enhanced sampling simulations are employed to investigate water-supercritical CO2 (scCO2) biphasic systems, with a focus on how interfacial properties govern HMF partitioning under varying temperature, pressure, and salt concentrations. MD simulations reveal that salt concentration has a stronger influence than pressure in modulating interfacial structure and properties, including interface thickness and interfacial tension. Despite an increased affinity of HMF for the scCO2 phase at elevated salt concentrations and pressures, it preferentially accumulates at the interface due to kinetic barriers and weaker stabilization in scCO2, as revealed by free-energy calculations using enhanced sampling simulations. Building on these insights, a strategy employing amphiphilic modifiers is used to enhance HMF extraction by facilitating its transport across the interface and by stabilizing HMF in scCO2. This approach is validated through simulations with isopropanol, which show a significant increase in HMF density in the scCO2 phase.
The present study aimed to determine the accuracy of machine learning in predicting soft tissue changes after orthodontic treatment after the extraction of premolars using frontal facial photographs. Pretreatment (T0) and posttreatment (T1) frontal photographs of 100 patients (70 females and 30 males) aged 17-28 years who underwent orthodontic treatment with premolar extraction were collected according to the inclusion and exclusion criteria. Images were digitized, and landmarks were annotated using the Visual Geometry Group image annotator (Visual Geometry Group, Department of Engineering Science, University of Oxford, Oxford, United Kingdom). A high-resolution network model was developed to extract and predict key points using the affine transformation technique. The mean difference between the actual and predicted models in the frontal measurements was assessed. The accuracy of the predicted model in assessing soft tissue changes was evaluated using the mean absolute error and the mean absolute percentage error. No statistically significant difference was found between the actual and predicted models in the soft tissue changes from T0 to T1. The overall accuracy of the predicted model in assessing soft tissue changes was 85.36%. However, the accuracy for predicting interlabial gap was notably lower (52.75%). Soft tissue treatment changes were predicted with moderate-to-high accuracy based on 2-dimensional frontal photographs, offering a noninvasive and clinically applicable solution. The high-resolution network model and 2-dimensional affine transformation technique ensured accuracy and reproducibility, highlighting the potential for artificial intelligence-driven automation in orthodontic diagnostics.
The present study explored whether people share a common understanding of different settlement concepts despite individual variation. Participants completed a property listing task where they were asked to generate features for 57 settlement concepts (e.g., "city," "college town"). We used hierarchical cluster analysis to identify distinct clusters based on shared features. Central tendencies extracted from the clusters at different levels of abstraction revealed featural prototypes and an overall family resemblance structure. To probe the effects of regional context on conceptual structure, we used a subset of participants who were long-term residents of Canada or the USA. We found that prototypical features varied regionally, suggesting an effect of participants' geographical region on conceptual structure. This work is the first to investigate how people represent settlements as concepts and helps centralize the utility of semantic feature norms in understanding how people collectively think about where they live, and the importance of context effects on representations of settlements.
Marketed cream formulations contain fatty acids as core constituents, required for its texture, uniformity, shelf life, and skin penetration. Given their importance in the cosmetic and pharmaceutical industries, real-time quantification and analysis of these compounds are crucial for assessing authenticity and ensuring compliance with regulatory specifications. In the present analysis, we employed gas chromatography with flame ionization detection (GC-FID) due to its sensitivity, robustness, and quantitative reliability. Overcoming the traditional method for fatty acid determination, we analyzed the sample without derivatization to fatty acid methyl esters (FAMEs). The derivatization step was initially omitted to simplify sample preparation, reduce analysis time and reagent consumption, and minimize the potential errors associated with incomplete esterification or degradation of unstable fatty acids during derivatization. Direct analysis was performed by dissolving the extracted sample in isopropyl alcohol and then filtering and injecting it into the GC-FID system under optimized chromatographic conditions. The optimized method was used for the concurrent quantification of lauric, myristic, palmitic, oleic, and stearic acids. The validation of the present work was executed in alignment with ICH Q2(R1) requirements. The method was further expanded to analyze real cream samples, thereby validating the accuracy of fatty acid profiling for routine quality control.
Cadmium (Cd) is a highly toxic heavy metal that accumulates in the kidney, causing oxidative stress, inflammation, apoptosis, and renal dysfunction. Ginger (Zingiber officinale) possesses antioxidant and anti-inflammatory properties, but comparative studies evaluating pre-treatment versus post-treatment regimens against cadmium nephrotoxicity remain limited. This study investigated the nephroprotective effects of aqueous Z. officinale rhizome extract in a comparative pre- and post-treatment design using histopathological, immunohistochemical, and molecular approaches. Thirty adult male Wistar rats were divided into five groups (n = 6): Control, CdCl₂ (5 mg/kg), Ginger alone (300 mg/kg), CdCl2 + Ginger post-treatment, and Ginger pre-treatment + CdCl2. Treatments were administered orally for six weeks after two weeks of acclimatization. Acute toxicity of the extract was evaluated using the Lorke method. Assessments included body weight, kidney morphometry, serum kidney function parameters, serum cadmium levels, relative gene expression of NF-κB and Nrf2, quantitative histopathology (H&E), and caspase-3 immunohistochemistry. Phytochemical profiling of the extract was performed using GC-MS and HPLC-DAD. The aqueous ginger extract showed high safety (LD50 > 5000 mg/kg). Cadmium exposure significantly reduced body weight gain, elevated serum creatinine and cadmium levels, upregulated NF-κB and Nrf2 expression, induced glomerular collapse and tubular damage, and increased caspase-3 expression (p < 0.05). Both pre- and post-treatment with ginger significantly improved body weight gain, reduced serum cadmium concentration, attenuated histopathological lesions, lowered caspase-3 immunoexpression, and modulated NF-κB and Nrf2 gene expression toward control levels. Notably, pre-treatment showed superior histological protection, unlike in other parameters. Aqueous Z. officinale rhizome extract effectively ameliorates cadmium-induced renal dysfunction through antioxidant, anti-inflammatory, anti-apoptotic, and metal-chelating mechanisms. Pre-treatment offered better structural protection, while both regimens provided significant functional and molecular benefits. These findings support ginger as a promising natural agent for mitigating heavy metal nephrotoxicity.
The development of sustainable food contact materials (FCMs) requires not only reduced environmental impact but also demonstrated safety. Cork-polypropylene composites (CPCs), incorporating unmatured cork and waste generated in cork stopper production, represent a promising alternative; however, their safety remains insufficiently addressed. This study provides a comprehensive safety assessment of CPCs containing 15% cork granulates derived from matured and unmatured cork. A multi-analytical approach combining morphological, thermal, and chemical characterisation (SEM, FTIR-ATR, GC-FID, GC-MS, LC-MS, ICP-OES) was integrated with migration testing under repeat-use conditions and sensory evaluation. Genotoxicity was assessed using the Ames bacterial reverse mutation test on migration extracts. Migration results showed low release of both polypropylene-related additives and cork-derived compounds, with furanic substances identified as process-related migrants but decreasing across repeated use cycles and remaining at low levels. No migration of high molecular weight cork constituents (triterpenoids, sterols) was observed in aqueous simulants. All quantified substances complied with regulatory limits. Ames test results demonstrated no mutagenic activity for any migration extract, confirming the absence of DNA-reactive hazards. Overall, CPCs exhibited stable structure, low sensory impact, and favourable chemical and toxicological profiles. This study provides new evidence supporting the safe use of cork-based composites in food-contact applications and advances the integration of bio-based materials within a circular economy framework.
A recent two-sample Mendelian randomization study suggested a possible causal association between hypothyroidism and selected sepsis types. We address this knowledge gap by examining the association between thyroid-stimulating hormone (TSH) levels and risk of sepsis and severe infectious diseases, using triangulation between traditional observational and Mendelian randomization (MR) analyses. Baseline characteristics and TSH-measurements were collected from adults (>20 years) at the time of participation in the prospective, population-based Trøndelag Health Study (The HUNT Study), and linked to hospital records for ascertainment of infectious diseases. Time-to-event analyses with Cox regression was used to assess the association between TSH levels and risk of sepsis, adjusting for confounders. Next, we extracted uncorrelated (R2 < 0.01) single-nucleotide polymorphisms strongly associated (p-value < 5e-8) with TSH levels from genome-wide association studies of European ancestry participants in the ThyroidOmics Consortium. Genetic associations with risk of sepsis were extracted from European ancestry participants in the UK Biobank. Secondary genetic analyses examined other measures of thyroid function (FT4, FT3, autoimmune thyroid disease and deiodinase activity) with sepsis risk, along with genetic and observational analyses of risk of lower respiratory tract infections (LRTI) and upper urinary tract infections (UUTI). In the observational analyses of 45,364 subjects in HUNT there was no association between baseline normal-range TSH and sepsis risk [HR 0.98 (95% CI 0.93-1.04) per mU/L unit increase]. TSH levels <0.5 mU/L was associated with higher sepsis risk (HR 1.50, 95% CI 1.19-1.90). In the MR analyses (271,040 subjects with TSH-measurements, and 10,154 cases with sepsis), there was no association between normal-range TSH and sepsis risk [OR 1.04 (95% CI 0.98-1.10), per SD increase]. Secondary analyses supported no link between thyroid function and risk of sepsis, LRTI or UUTI. Variation in baseline thyroid function in the general adult population does not causally influence the risk of sepsis, LRTI or UUTI. In contrast to earlier MR work based on genetic liability to overt hypothyroidism in clinical populations, our findings indicate that mild deviations in thyroid function within and around the reference range are unlikely to be useful targets for sepsis prevention or risk stratification.
Objective: To review conclusive evidence concerning the usage, efficacy, and side effects of 2 N-methyl-D-aspartate (NMDA) receptor antagonists, memantine and amantadine, in children with autism spectrum disorder (ASD) to determine their effectiveness in treating associated core and comorbid conditions. Data Sources: A comprehensive literature search was conducted across multiple electronic databases, utilizing a combination of controlled vocabulary and keywords: autism spectrum disorder, autistic disorder, Asperger's, PDD-NOS, neurodevelopmental disorders, memantine, amantadine, and NMDA receptor antagonist. Study Selection: The initial screening identified 87 studies, and 11 were selected for full review. A total of 8 studies met the inclusion criteria of the qualitative synthesis and were included in the systematic review. The inclusion criteria were children aged <18 years with an ASD diagnosis, studies assessing amantadine or memantine with a comparison of placebo or treatment as usual, studies reporting clinical improvement in common ASD symptoms, and clinical trials or retrospective chart reviews. Exclusion criteria included unpublished data and multiple reports from the same dataset. Data Extraction: Data were extracted using a standardized form focused on study design and validity domains. Multiple reviewers independently abstracted data, with discrepancies resolved by consensus. Data Synthesis: Memantine and amantadine demonstrated favorable safety profiles. However, the evidence regarding their efficacy in addressing core ASD symptoms was inconclusive. Memantine was associated with potential improvements in cognitive and behavioral outcomes, though the evidence was confounded by methodological limitations. Amantadine showed potential in adjunctive treatment with risperidone, particularly in mitigating disruptive behaviors, but variability in outcomes between parent-reported and clinician-assessed measures raised concerns about the reliability and robustness of the findings. Conclusions: This systematic review highlights the need for large-scale, multicenter, double-blind, randomized controlled trials with rigorously standardized assessment tools and diverse clinical populations to better define the therapeutic potential and establish guidelines for the use of memantine and amantadine in ASD. Prim Care Companion CNS Disord 2026;28(4):25r04152. Author affiliations are listed at the end of this article.
Time-domain neural beamformers typically extract pairwise microphone features and process them using sequential networks. While this approach can capture temporal dynamics of the input features containing spatial cues, it does not explicitly model spatial and temporal information jointly, which constrains the network's ability to learn rich spatio-temporal (ST) representations for multichannel speech enhancement. This paper proposes STBFNet, a time-domain neural beamforming network that leverages multi-scale ST feature learning. In the proposed model, local ST features are first extracted and aggregated through multi-scale convolutional operations, enabling the network to jointly capture temporal dependencies and channel-wise correlations. Simultaneously, spatial information compensation is employed to enhance global channel interactions. The resulting features are fused and further processed, where self-attention is utilized to learn long-range contextual dependencies, and a modified Conformer-style convolution is applied to strengthen ST feature integration. Through these designs, the network produces refined ST representations, leading to improved performance in time-domain beamforming. Experimental results demonstrate the superiority of STBFNet over existing multichannel filtering methods. The proposed STBFNet has a smaller model size and achieves lower latency while maintaining competitive performance.
Pistacia lentiscus L., commonly known as the mastic tree, is a Mediterranean plant recognised for its anti-inflammatory and antioxidant properties. Therefore, extensive research has focused on the chemical composition and biological activities of its gum, bark, leaves, and flowers. The chemical content of P. lentiscus is influenced by geographical location and soil conditions. In this study, ethyl acetate and hexane extracts of the bark and gum of Pistacia lentiscus L. collected from Çeşme (İzmir, Türkiye) were analysed using gas chromatography-mass spectrometry (GC-MS), and their antioxidant activities were evaluated. Total antioxidant capacity, radical scavenging activity, and total phenolic content of the extracts and branch distillates were determined using FRAP, CUPRAC, DPPH, and Folin-Ciocalteu assays. The findings indicate that Pistacia lentiscus samples from the Çeşme region display antioxidant activities comparable to those reported in previous studies.
Globally, cardiovascular diseases (CVDs) constitute a leading cause of death. Significant reductions in mortality are possible with early detection and treatment of CVDs. The electrocardiogram (ECG) is an easy-to-understand, non-invasive technique for assessing the state of the heart. A specific CVD can be determined by a slight deviation in the ECG, which is difficult and time-consuming to diagnose manually. This work suggests an empirical mode decomposition (EMD)-based approach together with time and frequency series feature extraction for the automated identification of CVDs. Five CVDs are taken into consideration in this work: myocardial infarction (MI), hypertrophic cardiomyopathy (HCM), dilated cardiomyopathy (DCM), bundle branch block (BBB) and coronary artery disease (CAD). The decomposition of signals using EMD yields amplitude- and frequency-modulated waves called intrinsic mode functions (IMFs). Temporal and spectral features are extracted from the obtained IMFs. These features are ranked using the one-way anova test, and the ranked features are fed as inputs to the classification model. When combined with eleven features, the extreme gradient boosting classifier allows the suggested approach to attain a maximum classification accuracy of 99.56%. On the proposed database and 92.4% on MIT-BIH database, demonstrating strong generalizability. Clinical validation on 6,55,659 beats from RELA Hospital yielded 81.3% accuracy, highlighting the challenges of real-world deployment.
The genus Plectranthus sensu lato is known to produce abietanes, which exhibit significant potency against Gram-positive bacteria. It is hypothesized that abietanes exert their antibacterial effects by disrupting microbial cell walls. Recent studies have classified Plectranthus s.l. abietanes into six distinct groups. However, the impact of this chemical diversity on the response of abietanes against Gram-positive bacteria remains unexamined. Therefore, a structure-activity relationship study on abietanes from species of Plectranthus sensu stricto and Coleus was conducted to identify active fragments or pharmacophores in abietane-type diterpenoids that could be responsible for their activity against the Gram-positive bacteria Staphylococcus aureus. The study followed a bio-guided approach and included chemical profiling of the extracts, performed using NMR spectroscopy, and comparative analysis, using LC-MS in both positive and negative ionization modes. Results revealed all six classes of abietanes occurred as major constituents of the extracts, with distinctive chemical markers differentiating between Plectranthus s.s. and Coleus species. Oxidation of the B-ring in acylhydrobenzoquinones was identified as a key factor contributing to the strong antibacterial activity of Plectranthus s.l. derived compounds against S. aureus. Analysis of 28 species of Plectranthus s.l. revealed four main clusters based on the accumulation of various abietane classes. The active acylhydrobenzoquinones were a defining factor in the separation of one of the groups of plants while another cluster in both Plectranthus s.s. and Coleus separated based on flavonoid contents. It would be of interest to examine how this trend might be broadened to encompass all species within each genus.
Contaminated food and water represent potential infection sources of Angiostrongylus cantonensis, a parasitic nematode with significant zoonotic potential. Limited environmental surveillance may be due to the absence of simple and reliable screening methods. Different methods of A. cantonensis DNA detection were evaluated on lamb's lettuce (Valerianella locusta). First, three methods for concentrating larvae from water samples spiked with 1 and 10 third-stage larvae (L3) were compared: simple sedimentation and two sieving techniques. Then, two DNA extraction modifications were assessed on spiked lettuce samples. Lastly, the detection limit was evaluated using lettuce samples spiked with 1, 5, 10, 20, and 50 L3. Method efficiency was assessed using a sensitive quantitative polymerase chain reaction (qPCR). The highest DNA yields were obtained using a sieving method with a direct sieve DNA extraction employing glass beads and chelating resin. Using this approach, the detection limit was determined to be a single L3 in 50 g of lamb's lettuce. Detection was successful from both the sieve-retained material and the sedimented flow-through. Sieving techniques provide a promising screening approach for detecting larval stages of parasites, such as A. cantonensis in vegetables. Combined with a sensitive qPCR, detection was consistently achieved at 1 L3 under controlled conditions. Detection of parasite DNA in environmental samples indicates a potential health risk and supports the applicability of this method for food safety monitoring, with possible adaptation to other vegetables.
Introduction Lack of health insurance is known to delay care-seeking and contribute to preventable morbidity. Although Japan has established a universal health insurance system, more than 10% of households have unpaid insurance premiums, and approximately 2.5% possess insurance certificates with restricted or no validity. Under these circumstances, preventable deaths attributable to delayed medical care for financial reasons have been reported. Individuals facing unstable employment or job loss are particularly vulnerable to premium non-payment, even when willingness to pay exists. The actual scale, characteristics, and clinical outcomes of uninsured patients in Japan remain largely unknown. Materials and methods We conducted a retrospective chart review at a small suburban hospital in Japan. The characteristics of patients without any form of health insurance at the time of the encounter were extracted from medical records from October 2021 to September 2025. Patient consent was obtained on an opt-out basis. Results Over the four-year study period, 20 patients received medical care at the hospital without any form of health insurance. Twelve patients (60%) required hospital admission at their first encounter. Nine patients (45%) were diagnosed with psychological disorders. Severe functional impairment or death occurred in 10 patients (50%), including two cases of death. Prior to hospital presentation, four patients (20%) had attempted to apply for public assistance at a municipal office but were denied. Conclusions Some individuals who remained outside the national insurance system present with severe and preventable illnesses. Although this study cannot establish a causal relationship, lack of insurance coverage may contribute to delayed care and adverse health outcomes. Efforts to remove administrative barriers and strengthen social work support may help reduce preventable harm and promote health equity within Japan's healthcare system.
Prostate cancer (PCa) remains a leading cause of cancer-related mortality in men, with current treatments often limited by drug resistance and systemic toxicity. Although traditional Chinese medicine components such as Astragaloside IV and polypeptide extract from scorpion venom (PESV) have demonstrated promising antitumor activity, their clinical translation is hampered by poor bioavailability and lack of tumor specificity. To address these limitations, we engineered an E3 aptamer-modified T cell-derived exosomal nanoplatform (EAPE) for the targeted co-delivery of Astragaloside IV and PESV in prostate cancer therapy. EAPE was constructed and characterized, and its targeting capability, biosafety, and therapeutic performance were evaluated in vitro and in vivo. In vitro, the antitumor efficacy was assessed by proliferation, migration and apoptosis assays, while the immunomodulatory effects were investigated using a co-culture system of LNCaP cells and T lymphocytes. In vivo, the antitumor efficacy and immune activation were examined in prostate cancer xenograft mouse model, with tumor growth inhibition, apoptosis and immune responses measured. EAPE demonstrated efficient tumor-targeting capability and favorable biosafety profiles both in vitro and in vivo. EAPE demonstrated superior therapeutic efficacy against PCa by inhibiting proliferation and migration of prostate cancer cells and inducing apoptosis, while suppressing immunosuppression and activating antitumor immune response. This study presents a biologically derived, targeted nanodelivery system that improves the delivery efficiency and therapeutic efficacy of Astragaloside IV and PESV. These findings support the potential of exosome-based nanoplatforms as promising strategies for enhancing the translational application of traditional Chinese medicine-derived therapeutics in prostate cancer.