As the threat of antimicrobial resistance (AMR) escalates globally, the influence of indiscriminate antimicrobial use (AMU) in livestock cannot be overlooked. Antimicrobial use practices are continually explored in larger food-producing animals; however, small ruminants (sheep and goats) receive comparatively less research attention. Our study addresses this gap by investigating small ruminant production practices in Nigeria and exploring how they affect the use of antimicrobials and alternatives. We adopted a mixed-methods study design. A semi-structured questionnaire was administered to 785 farmers. Following this, a focus group discussion (FGD) was conducted with 23 small ruminant industry stakeholders, which included farmers, para-veterinarians, and butchers. Participants were split into three round tables, with 7-8 participants per table. Of questionnaire respondents, 68% of farmers never vaccinated their flock against peste de petits ruminants (PPR) nor contagious caprine pleuropneumonia virus. Several health problems were regularly experienced by animals, including PPR, mastitis, and dermatophilosis. Diseases were mostly self-managed with antibiotics and herbs (> 70%) rather than through reliance on veterinary care (< 15%). More farmers (48%) used antibiotics than herbal remedies (14%) over the previous three months. Farmers' use of herbs was affected by their having low awareness of available options and how to use them appropriately. Perceived effectiveness also influenced farmers' choice between antibiotics and herbs, while economic considerations also led them to sell off sick animals before or during treatment. Among farmers who used animal health services, more farmers (59%) consulted unlicensed para-veterinarians and drugstore vendors rather than licensed government and private veterinarians (36%), a disparity attributed to the unavailability of qualified veterinary doctors. Most farmers had poor knowledge (62%), attitudes (47%), and practices (43%) towards AMU and AMR. We recommend conducting further studies to identify and investigate the efficacy of currently used herbs in treating common diseases. There is a crucial need to improve farmers' access to vaccines, veterinary care, and laboratory diagnostics. Barriers that hinder better compliance with regulations that govern the use of non-prescribed antimicrobials should be explored. Awareness programmes could be conducted to improve farmers' awareness of AMR and appropriate disease preventive practices.
Breast cancer exhibits profound molecular and clinical heterogeneity, manifested through diverse subtypes, metastatic patterns, and therapeutic responses. Emerging evidence underscores that this complexity stems not merely from genetic and epigenetic aberrations but also from post-transcriptional regulatory mechanisms, particularly alternative splicing (AS), which amplifies proteomic diversity and functional versatility. AS generates multiple mRNA isoforms from a single gene, directly influencing oncogenic processes including cell migration, metabolic reprogramming, immune evasion, and therapy resistance. Recent advances in high-throughput sequencing and bioinformatics have enabled systematic mapping of AS landscapes in breast cancer. This review delineates the mechanistic underpinnings of AS regulation and synthesizes cutting-edge discoveries in AS-driven oncogenic pathways. We further evaluate the translational potential of AS signatures in clinical applications, such as molecular subtyping, prognostic biomarkers, and splice-switching therapeutics. Finally, we address persisting challenges, including tumor heterogeneity and the lack of splicing specificity, that must be overcome to harness AS modulation as a precision oncology strategy.
The unfolded protein response (UPR) is a critical adaptive program triggered upon cellular stresses that profoundly reshapes the transcriptome and translatome. In the very first minutes of cellular stress, translation blockage, RNA decay and RNA granules formation prompt the synthesis of proteins essential to the stress response. Due to the dynamic nature of these processes, investigating translation upon stress has proven to be challenging; therefore, our understanding of these mechanisms and translatome rewiring upon stress remains limited. Here, we exploit O-Propargyl-puromycin (OPP) labelling of de novo peptides followed by LC-MS/MS to identify de novo proteins translated upon endoplasmic reticulum (ER) stress. Combined with transcriptomic analyses, our approach reveals that ER stress profoundly impacts the synthesis of core splicing factor proteins leading to a significant reshaping of the splicing landscape. We identify a signature of seven splicing events consistently occurring in mammalian cells exposed to ER stress. Using pharmacological, genetic, phosphoproteomic and sequencing approaches, we demonstrate that this specific signature is driven by PERK activation and is dependent on the axis CLK1/SRSF1. Our findings identify PERK/CLK1/SRSF1 -mediated splicing regulation as a new facet of ER stress, defining an ERi-splice signature spanning healthy and malignant tissues.
Resistance to osimertinib in lung adenocarcinoma presents a significant hurdle in contemporary lung cancer therapy, with splicing dysregulation being instrumental in tumorigenesis and progression. The mechanism via which alternate splicing facilitates osimertinib resistance in lung cancer is still ambiguous. We aimed to examine the pivotal function of the splicing factor hnRNPA1 in osimertinib resistance in lung cancer. The impact of hnRNPA1 on osimertinib resistance in lung cancer was confirmed by small interfering RNA and CDX models. RNA-seq, RIP, CLIP-qPCR, and COIP methodologies were utilized to examine the mechanistic role of hnRNPA1 in osimertinib resistance in lung cancer. Furthermore, virtual docking was employed to evaluate natural small-molecule drugs that target hnRNPA1.Elevated levels of the splicing factor hnRNPA1 were detected in lung adenocarcinoma cells exhibiting resistance to osimertinib, and the silencing of hnRNPA1 enhanced the sensitivity of these resistant cells to osimertinib. Mechanistically, hnRNPA1 governs multiple splicing events linked to cancer, notably the splicing of NEDD4L. Decreased hnRNPA1 enhances the generation of short NEDD4L splice variants, resulting in the ubiquitination and degradation of EGFR, which sensitizes resistant cells to osimertinib. Moreover, hnRNPA1 undergoes methylation modification by PRMT7, which facilitates splicing activities. The research elucidated the mechanism responsible for aberrant splicing in osimertinib resistance in lung cancer and identified hnRNPA1 as a prospective therapeutic target for counteracting this resistance.
The sensory properties of alternative proteins are key to consumer acceptance, yet the processes shaping their odor remain unclear. Solid-state fermentation (SSF), a promising method for producing alternative proteins from agro-industrial by-products such as bagasse, brans, pomaces, husks and oil cakes, is used in this study to model odor profile development of surplus bread crusts, supplemented with perennial ryegrass protein with Rhizopus oligosporus, Aspergillus oryzae, and Neurospora intermedia at 32 °C for up to 72 h. Volatile organic compounds (VOCs) were analyzed by solid-phase microextraction (SPME) followed by gas chromatography-mass spectrometry (GC-MS), identifying over 150 compounds. A mechanistic model based on the Weber-Fechner law predicted odor profiles from VOC concentrations, odor descriptors and thresholds, and was validated against a quantitative descriptive analysis (QDA) performed by a trained panel using multiple factor analysis (MFA). The model reflected changes in the overall odor intensity and sweet, baked, and grass-like notes, though correlations were weaker for fungal-derived descriptors (fruity, earthy, herbal). These findings elucidate how fungal SSF alters odor profiles in alternative proteins and establish a framework for mechanistic odor prediction in food systems.
Chronic hepatitis D is the most severe form of viral hepatitis. Despite recommendations for systematic screening of HBsAg-positive individuals, hepatitis D infection remains underdiagnosed. Standard HDV virological markers are classically assessed in serum or plasma specimens obtained through venous whole blood sampling. This approach is costly and challenging in resource-limited settings. Dried blood spot (DBS) sampling offers a practical alternative for large-scale screening, diagnosis, and monitoring of viral hepatitis. The goal of the study is to evaluate the performance of commercially available HDV diagnostic assays using DBS. Plasma and DBS-collected whole blood specimens from individuals chronically infected with HBV or coinfected with HBV and HDV were analysed for HDV antibody detection, HDV RNA quantification, HBsAg levels, and HDV genotyping. HDV antibodies were reliably detected in DBS after threshold adjustment, with the LIAISON XL Murex Anti-HDV assay showing superior sensitivity (99.3%) compared to HDV Ab DIA.PRO (90.3%), while both assays exhibited excellent specificity. HDV RNA was quantifiable in 86.8% of DBS specimens from patients with active infection, although levels were about 1.5 log10 lower than plasma, showing a strong correlation (r = 0.786; p < 0.0001). HBsAg was detectable and quantifiable in all DBS samples, correlating closely with plasma values (r = 0.98). DBS-based HDV genotyping was successful in 84.8% of samples and concordant with plasma results. DBS collection of whole blood from DBS appears to be a promising and practical alternative to conventional venous blood sampling for HDV screening, diagnosis, and monitoring. DBS sampling is a valuable tool for the large‐scale screening, diagnosis, and monitoring of viral hepatitis B and C. HDV virological markers can be easily detected in the whole blood. Collecting whole blood on filter paper offers a practical approach that could significantly contribute to the WHO's goal of eliminating viral hepatitis.
Strict environmental regulations require the production of ultra-low sulfur diesel (ULSD); however, the conventional hydrodesulfurization (HDS) process remains highly energy-intensive. Photocatalytic oxidative desulfurization (PODS) at room conditions has emerged as a promising green alternative. However, industrial applications of UV-based PODS are severely hampered by the Inner Filter Effect (IFE), in which the aromatic-rich diesel matrix strongly absorbs UV radiation, leading to severe photon starvation within the liquid volume and strictly limiting the reaction to the interface. This critical review highlights a paradigm shift towards visible- and near-infrared (red and NIR) light-driven PODS to overcome this optical deadlock. Although red light has excellent penetrating power on diesel matrices, its low photon thermodynamic energy (2.0 eV) fundamentally hinders direct C-S bond breaking, which actually demands a much higher activation energy. To bridge this energy deficit, advanced nonlinear photon manipulation strategies are comprehensively evaluated. This review shows that simultaneous Two-Photon Absorption (TPA) is the most superior mechanistic pathway, fundamentally outperforming Second Harmonic Generation (SHG) and Triplet-Triplet Annihilation (TTA). While simultaneous TPA utilizing sensitizers with giant absorption cross-sections (> 1000 GM) provides direct access to highly excited states (Sn or Q1), it fundamentally requires high-intensity laser excitation. Alternatively, defect-mediated Two-Step Two-Photon Absorption (TS-TPA) exploits real intermediate states to bridge the energy gap, triggering highly efficient photochemical degradation at highly accessible, low excitation densities (~ 0.1 W cm- 2). Finally, future prospects emphasizing continuous-flow microreactors and advanced optical encapsulation strategies are discussed to realize commercially viable red-light PODS.
Hydroxyurea (HU) is a first-line oral cytoreductive agent for selected patients with myeloproliferative neoplasms (MPNs), including polycythemia vera (PV), due to its efficacy and tolerability. Although cutaneous ulceration is a recognized complication of long-term HU exposure, HU-associated digital gangrene is rare, and upper-extremity involvement in PV has not previously been reported. We describe the first case of HU-associated digital gangrene in a patient with PV. A 72-year-old man with well-controlled Janus kinase 2 (JAK2)-positive PV, treated with HU for more than a decade, developed painless dry gangrene of the right index and middle fingers. Vascular imaging showed no significant arterial occlusion or embolic source. Hematologic parameters remained within target ranges, and the workup for autoimmune disease, infection, and hypercoagulability was unremarkable. HU was discontinued, and the ischemia stabilized without surgical intervention. With no alternative etiology identified, delayed HU-associated vasculopathy was suspected. Our literature review identified three previously reported cases of HU-associated digital gangrene, though limited to the lower extremities - two in chronic myeloid leukemia (CML) and one in sickle cell disease (SCD). In each case, gangrene developed after prolonged HU exposure, alternative etiologies were not substantiated, and stabilization or clinical improvement followed HU withdrawal. The present case aligns with this pattern while extending the reported phenotype to well-controlled PV and upper-extremity digits. Given the small number of reported cases, the pathophysiology remains incompletely defined and is largely extrapolated from studies of more frequently described HU-associated ulceration, histopathologic reports of HU-related tissue injury, and in vitro studies of HU effects on endothelial and circulating cells. Plausible mechanisms include cumulative endothelial injury, localized thrombo-occlusive microvascular dysfunction, impaired vascular and cutaneous repair, and interaction with PV-related microvascular susceptibility. Clinicians should include HU-associated vasculopathy in the differential diagnosis of otherwise unexplained digital ischemia, as prompt drug cessation may limit progression and improve digit salvage.
To compare perioperative outcomes of robotic-assisted thoracic surgery (RATS) versus video-assisted thoracoscopic surgery (VATS) for mediastinal tumor resection in a propensity score-matched cohort. This retrospective study analyzed patients undergoing minimally invasive mediastinal tumor resection between January 2022 and November 2025. Patients were categorized into RATS and VATS groups. Propensity score matching balanced baseline characteristics including age, gender, body mass index, ASA score, Charlson Comorbidity Index, tumor size, tumor location, pathology, and prior thoracic surgery history. The primary outcome was postoperative complications of Clavien-Dindo grade ≥ II. Secondary outcomes included operative time, blood loss, chest tube duration, hospital stay, costs, and pain scores. After matching, 102 patients (51 per group) were included. RATS was associated with significantly less intraoperative blood loss [median difference - 15 mL (95% CI: - 20 to - 10), P < 0.001], shorter chest tube duration [median difference - 1 day (95% CI: - 1.7 to - 0.3), P = 0.003], and shorter hospital stay [median difference - 1 day (95% CI: - 1.7 to - 0.1), P = 0.009]. Total costs were higher in the RATS group [median difference +$4,400 (95% CI: $3,800-$5,000), P < 0.001]. Operative time, postoperative pain scores, and complication rates (grade ≥ II: 9.8% vs. 13.7%, P = 0.55) were comparable between groups. Using an alternative definition (grade ≥ III), complications occurred in 3.9% (RATS) vs. 5.9% (VATS), P = 0.65. For mediastinal tumor resection, RATS offers modest perioperative benefits including reduced blood loss and shorter recovery times compared to VATS, but at substantially higher cost and without a reduction in major complications. RATS is a feasible and safe minimally invasive alternative, but its advantages over VATS are small and must be weighed against economic considerations. Routine adoption is not supported by the current evidence; selection should be individualized.
Short dental implants (SDIs) have emerged as a promising, less invasive alternative to conventional-length implants (CLDIs) for oral rehabilitation in patients with limited bone height, reducing the need for additional surgical procedures while maintaining functional and aesthetic success. This systematic review and meta-analysis aimed to answer: "Do single crowns supported by SDIs have a clinical success rate comparable to (or non-inferior to) that of CLDIs?", focusing on survival rates, periodontal health, marginal bone loss (MBL), and biological complications. A comprehensive search was performed in Embase, Lilacs, PubMed/MEDLINE, Science Direct, Scopus, Google Scholar, and ProQuest, including all available years. Randomized and non-randomized clinical trials comparing single crowns supported by SDIs and CLDIs were included. Studies had to assess survival, periodontal health, MBL, and biological complications. Retrospective, non-comparative, and unspecified designs, reviews, and case reports were excluded. Screening and selection were conducted through titles, abstracts, and full-text reviews. Following PRISMA guidelines, data extraction and quality assessment were performed. The review was registered in PROSPERO. Risk of bias was evaluated using RoB2 and ROBINS-I, and meta-analyses were conducted with random-effects models, with a statistical significance of p < 0.05. The outcomes evaluated were implant survival, MBL, and probing depth (PD), measured by standardized mean differences (SMD) and relative risks (RR) with 95% confidence intervals (CIs). A total of 1146 potential studies were identified; 626 were screened, 30 fully reviewed, and 17 included. All studies showed a high risk of bias. SDIs presented survival rates comparable to CLDIs in both splinted (RR = 0.98; 95% CI = 0.96-1.00; p = 0.05) and non-splinted single crowns (RR = 0.96; 95% CI = 0.91-1.00; p = 0.07). MBL showed a small but statistically significant reduction for both (non-splinted: SMD = - 0.11; p = 0.03; splinted: SMD = - 0.41; p = 0.01), with lower PD in non-splinted crowns (SMD = - 0.64; p = 0.03). This systematic review with meta-analysis suggests that SDIs achieve clinical outcomes comparable to, and not inferior to, CLDIs for splinted or non-splinted single crowns regarding implant survival up to 10 years. Despite methodological limitations, SDIs may represent a viable and less invasive treatment alternative.
Antineutrophil cytoplasmic antibody (ANCA) associated vasculitides (AAV) are rare, life-threatening autoimmune conditions requiring complex pharmacological management. Contemporary strategies have transitioned from empirical cytotoxic therapy toward mechanistically targeted approaches, reflecting a deeper understanding of B-cell immunity and the alternative complement pathway. This review synthesizes current guidance and presents recent evidence on treatment modalities and emerging options. This paper reviews and synthesizes current international guidance from the European League against Rheumatism (EULAR), British Society for Rheumatology (BSR), and the American College of Rheumatology (ACR) on remission induction and maintenance for granulomatosis with polyangiitis (GPA), microscopic polyangiitis (MPA), and eosinophilic granulomatosis with polyangiitis (EGPA). Key shifts discussed include the established role of rituximab as a primary induction agent, the implementation of reduced-dose glucocorticoid tapering, and the integration of anti-IL-5 therapies for eosinophilic phenotypes.Expert opinion: In our opinion, the management of AAV has entered a precision era defined by targeted, steroid-sparing regimens. The most significant progress lies in standardizing B-cell depletion and the emergence of complement inhibition. Despite the evolution of steroid-sparing protocols, mitigation of treatment-related morbidity remains a primary challenge. Ultimately, AAV management must transition toward biomarker-driven precision medicine to individualize therapy and improve long-term outcomes.
Since real-world data lack information on treatment assignment (usually the only information available is treatment prescription/administration) and early outcomes can happen in advanced cancer, some design choices may introduce immortal time due to selection or due to treatment strategy misclassification. To illustrate design choices that would biassedly introduce immortal time and propose unbiased alternatives, using as a case study the estimation of the effects of different monoclonal antibody (mAb) sequencing strategies on overall survival in colorectal cancer patients. We specified a target trial to estimate the effect on overall survival of initiating mAb within 8 weeks of starting first line chemotherapy versus initiating mAb with second line chemotherapy. The first biased design choice would be requiring the initiation of second line chemotherapy for eligibility. This was avoided by aligning eligibility with the initiation of first line chemotherapy. The second biased design choice would be excluding from the first line mAb strategy those who die during the grace period without starting mAb. This was avoided separately via cloning, censoring and weighting and by sequential emulation. There were 1014 eligible patients in GEMCAD 1401. The CCW approach estimated a 4-year survival difference (second-first line mAb) of 4.3% (95% CI) (-3.3%; 10.0%) and a risk ratio (RR) of 0.95 (0.88; 1.04). Sequential trial emulation estimated a survival difference of -2.8% (-7.5%; 1.2%) and a RR of 1.04 (0.98; 1.10). Biased analyses that introduced immortal time estimated an implausible early survival benefit for mAb initiation with second-line (6-month RR of 0.13 (0.03; 0.28)). Our results are compatible with no effect on survival by mAb sequencing. Misalignment of time zero, treatment assignment, and eligibility introduced immortal time.
Heart failure remains a leading cause of morbidity and mortality worldwide, and current therapies largely focus on symptom management and slowing disease progression rather than correcting the underlying molecular abnormalities. Recent advances in genome editing technologies have created new opportunities to treat heart failure. Among these approaches, CRISPR-Cas9 base editing has emerged as a particularly promising strategy because it enables precise nucleotide conversions without introducing double-strand DNA breaks and demonstrates relatively high efficiency in vivo. While correction of disease-causing mutations by CRISPR-Cas9 base editing represents an important application of genome editing, an alternative strategy is to directly modulate key signaling pathways that drive cardiac dysfunction. Protein kinase Cα (PKCα) functions as a key regulator of cardiac contractility and pathological remodeling. Precision editing of phosphorylation sites that control PKCα stability or activation may therefore represent an effective strategy to suppress maladaptive kinase signaling in cardiomyocytes. This concept of "precision signaling modification" may provide a broadly applicable therapeutic approach for heart failure. Similar strategies may also be applicable to other signaling molecules, including Ca2+/calmodulin-dependent protein kinase IIδ (CaMKIIδ), and illustrate the broader potential of signaling-focused genome editing approaches. Despite these advances, several challenges remain for clinical translation, including efficient delivery of genome editing components to the adult heart, long-term safety, and potential immune responses. Continued advances in delivery technologies and genome editing platforms may ultimately enable durable, potentially one-time therapeutic interventions for heart failure.
The traditional Medicine Ball Throw (MBTT) for assessing upper-extremity explosive strength inherently favours heavier and taller participants. We designed three alternative indices to adjust for this anthropometric bias: (1) multiplying MBTT by the height-to-mass ratio (MBTA1), (2) dividing MBTT by the product of mass and height (MBTA2), and (3) dividing MBTT by the body mass index (MBTA3). Using age- and sex-adjusted LMS-based z-scores from two independent cohorts (821 children, aged 8.3 ± 0.7 years; 354 adolescents, aged 14.4 ± 2.2 years), we compared the variants' alignment with the concave BMI-profiles of five standard fitness tests (6-minute run, standing long jump, jump sideways, 4 × 10 m shuttle run, push-ups). The three anthropometrically enriched variants yielded significantly better goodness of fit than MBTT with MBTA1 performing best overall. Furthermore, when deployed as independent covariates, the enriched indices demonstrated systematically higher predictive power for standard fitness tasks than the unadjusted distance. We recommend adjusting MBTT via the height-to-mass ratio for a fairer assessment, though its structural overlap with BMI requires caution in predictive models.
Intravenous polymyxin B (PMB) is widely used for treating drug-resistant gram-negative pneumonia, though its efficacy in pulmonary infections is considered limited. This study aims to identify patients with severe pulmonary infections suitable for treatment with intravenous PMB alone and to recommend individualized optimal dosing regimens. Physiologically based pharmacokinetic (PBPK) models, developed using data from mice, healthy subjects, and critically ill patients, were employed to predict PMB concentrations in epithelial lining fluid (ELF) and plasma. All models were validated against published pharmacokinetic data, and Monte Carlo simulations were used to evaluate various dosing regimens. The therapeutic target was defined as an ELF steady-state 24 h area under the concentration-time curve to minimum inhibitory concentration (AUCss,24h/MIC) ≥ 50. Simulations showed that intravenous PMB maintenance regimens of 100 mg, 1.25 mg/kg, or 1.50 mg/kg q12h enabled 94.4%, 82.2%, and 93.5% of critically ill patients to achieve the ELF pharmacokinetic/pharmacodynamic (PK/PD) target for pathogens with an MIC of 1 mg/L. A plasma AUCss,24h/MIC threshold of 80.6 was identified as an alternative PK/PD target, with retrospective analysis confirming significantly better outcomes for patients exceeding this threshold (p = 0.038), warranting prospective validation. Overall, PBPK modeling supports 100 mg or 1.50 mg/kg q12h as candidate initial intravenous regimens for critically ill patients with pulmonary infections. Dosing adjustments based on MIC and plasma AUCss,24h/MIC value are recommended. Adjunctive nebulization or combination therapy is advised for MICs ≥ 2 mg/L, while 50 mg q12h is sufficient for MICs ≤ 0.5 mg/L. Plasma AUCss,24h/MIC values exceeding 80.6 may guide individualized dosing when therapeutic drug monitoring is available.
The global transition toward sustainable agricultural systems has intensified the search for fungal-based biopesticides, such as Trichoderma asperellum, to replace synthetic pesticides. Although considered safer, these products are not biologically inert and may elicit cellular responses in non-target organisms, making comparative ecotoxicological evaluation with conventional pesticides, such as mancozeb and abamectin, essential. This study aimed to evaluate and compare the cytogenotoxic effects, nucleolar alterations, and phytotoxicity of a T. asperellum-based biopesticide in relation to commercial formulations of mancozeb and abamectin. A multi-endpoint experimental design was applied, integrating the Allium cepa bioassay (mitotic index, chromosomal aberrations, micronuclei, AgNOR, plasma membrane integrity, electrolyte leakage, and TTC-based metabolic/mitochondrial activity) with seedling phytotoxicity assays in monocotyledonous (Triticum aestivum, Sorghum bicolor) and dicotyledonous (Raphanus sativus, Lactuca sativa) species. Treatments were tested at label-based concentrations: 2.0 g L-1 T-Protec® (0.07 g a.i. L-1), 13 g L-1 Manzate WG® (9.75 g a.i. L-1 mancozeb), and 1 mL L-1 Vertimec 18 EC® (0.018 g a.i. L-1 abamectin), under continuous exposure (A. cepa for 92 h; seedlings for 72 h for root assessment and 96 h for shoot assessment). Although all treatments significantly reduced the mitotic index and altered membrane integrity, toxicity patterns diverged considerably. The T. asperellum-based biopesticide showed lower genotoxicity, absence of micronucleus induction, and preservation of mitochondrial activity. In contrast, mancozeb and abamectin induced chromosomal anomalies and dysregulated the cell-cycle, reduced mitochondrial activity, and inhibited root and shoot development in the tested seedlings. Overall, despite inducing measurable cellular responses, the biopesticide showed substantially lower cytogenotoxicity and phytotoxicity than conventional synthetic pesticides, potentially making it a safer alternative.
Aluminum adjuvants have been used in licensed human vaccines for almost a century. However, they can neither induce an efficient cellular immune response, nor be frozen or lyophilized due to aggregation and diminished adjuvant efficacy. This study developed a series of lyophilizable nano-aluminum adjuvants by coating nanoscale aluminum adjuvant (Adju-Phos) with glycol chitosan (GCS, 20 kDa) using microfluidics technology. When the GCS/alum weight ratio was higher than 4:1, the nano-aluminum adjuvants withstood autoclaving and freeze-drying treatments in the presence of trehalose without significant physicochemical changes. Moreover, the lyophilized nano-aluminum adjuvants remained stable for 15 months at room temperature. When combined with HPV16 L1 virus-like particles (VLPs), the nano-aluminum adjuvant with 7:1 GCS/alum weight ratio (G7-A1) elicited stronger adaptive immunity, as evidenced by higher neutralizing antibodies, balanced IgG1/IgG2c ratio, elevated IFN-γ/TNF-α/IL-4/IL-10 secretion, and increased Th1/CTL/memory T cell frequencies. Furthermore, combining G7-A1 with MF59-like emulsion or C-di-AMP enhanced antigen-specific humoral and cellular immunity, memory T cell responses, and dendritic cell activation. These findings demonstrate the great potential of nano-aluminum adjuvants in improving vaccine efficacy, and suggest that they may serve as alternatives suitable for vaccines requiring balanced Th1/Th2 immunity or strong cellular immunity.
Gastric carcinoma remains a major cause of cancer-related mortality worldwide, underscoring the need for alternative therapeutic strategies. Zinc oxide nanoparticles (ZnO NPs) have attracted attention in cancer research due to their ability to generate reactive oxygen species (ROS), relative cost-effectiveness, and suitability for green synthesis approaches. In this study, ZnO NPs were synthesized via an eco-friendly biogenic method using Artemisia sieberi, and their anticancer-related effects were evaluated in vitro. ZnO nanoparticles were synthesized using an aqueous extract of Artemisia sieberi and characterized by FE-SEM, DLS, EDS, and XRD. Their biological effects were investigated in human gastric adenocarcinoma (AGS) cells and normal human embryonic kidney (HEK-293) cells. Cytotoxicity was assessed using the MTT assay, apoptosis by flow cytometry, cell migration by a wound healing assay, and gene expression by qRT-PCR. Statistical analysis was performed using one-way ANOVA, with p < 0.05 considered significant. Physicochemical characterization confirmed the formation of spherical to polyhedral ZnO nanoparticles with sizes ranging from 37 to 122 nm. The nanoparticles exhibited concentration-dependent cytotoxicity in AGS cells, with an IC₅₀ of approximately 480 µg/mL, compared to an IC₅₀ of approximately 1250 µg/mL in HEK-293 cells, corresponding to a modest selectivity index (~ 2.6). Flow cytometric analysis demonstrated increased apoptosis in AGS cells, with up to 61.6% apoptotic cells observed at the highest concentration. In addition, ZnO NPs significantly inhibited cancer cell migration. Molecular analysis revealed significant upregulation of p53, Caspase-9, and Caspase-3, suggesting activation of the intrinsic mitochondrial apoptotic pathway. The findings indicate that Artemisia sieberi-derived ZnO nanoparticles can induce apoptosis and inhibit migration in gastric cancer cells in vitro, potentially through a p53-mediated mitochondrial mechanism. Although the observed selectivity toward cancer cells was limited, these results support the preliminary potential of biogenically synthesized ZnO nanoparticles as candidates for further investigation. Additional studies, including in vivo models and broader mechanistic analyses, are required to clarify their therapeutic relevance.
SMARCA4-deficient uterine tumours encompass two distinct entities: SMARCA4-deficient undifferentiated/dedifferentiated endometrial carcinoma (SDUDEC) and SMARCA4-deficient uterine sarcoma (SDUS). Despite their divergent classifications, these tumours share overlapping clinicopathological and molecular features that complicate diagnosis. This study characterises their distinguishing features and clinical significance through analysis of 34 SDUDEC and 14 SDUS cases, integrating clinicopathological, immunohistochemical [BRG1 (encoded by SMARCA4), BRM (encoded by SMARCA2), INI1 (encoded by SMARCB1), ARID1A, ARID1B, SOX2, claudin-4, CK8/18, Chromogranin, synaptophysin, INSM1, p53, and MMR proteins], and next-generation sequencing (NGS; 14 SDUDEC and 14 SDUS) data. SDUDEC patients were significantly older than SDUS patients, with a median age of 54 years (range 28-63) compared to 37 years (range 24-58) (p<0.01). SDUDEC exhibited marked immunohistochemical heterogeneity, whereas SDUS showed uniform profiles: all SDUS cases (10/10) demonstrated dual BRG1/BRM loss with preserved expression of other SWI/SNF complex proteins, absent claudin-4 (0/12) and SOX2 (0/8) expression, and no MMR deficiency (0/14) or mutant-type p53 (0/13). NGS revealed frequent endometrial carcinoma-associated alterations in SDUDEC but rarely in SDUS. Prognostic outcomes did not differ significantly (p>0.05). These findings highlight distinct molecular landscapes: SDUDEC aligns with endometrial carcinogenesis, while SDUS may arise via alternative SMARCA4-dependent mechanisms. A diagnostic algorithm combining endometrial carcinoma molecular classification, SWI/SNF protein testing, and thorough sampling is proposed. This study expands the clinicopathological and molecular spectrum of SMARCA4-deficient uterine tumours, underscoring the need for entity-specific management strategies.
To examine how changes in Medicare payments and industry transfers of value (TOVs) are associated with changes microinvasive glaucoma surgery (MIGS) procedure rates. Retrospective, observational cohort study. Ophthalmologists who performed glaucoma surgery for Medicare beneficiaries from 2018-2023. A 20% nationally representative sample of Medicare Part B Carrier claims from 2018-2023 identified trabecular meshwork bypass stent implantation, goniotomy, canaloplasty, and trabeculectomy procedures using Current Procedural Terminology codes. Procedures were aggregated at the physician-year level using National Provider Identifiers. Medicare payment changes for trabecular bypass stents implantation implemented on January 1, 2022 were examined alongside contemporaneous industry stent-related TOV payments from trabecular meshwork bypass stent manufacturers (Alcon and Glaukos), obtained from the Centers for Medicare & Medicaid Services Open Payments database. Annual and quarterly procedure volumes were compared before (2018-2021) and after (2022-2023) the policy change. Trabeculectomy served as a control procedure. Annual procedure volumes for trabecular stents, goniotomy, canaloplasty, and trabeculectomy; mean number of procedures performed per physician; association of CMS payment rates and industry TOV payments with individual procedure rates. Following a Medicare payment reduction for trabecular stents from approximately $500 to $34 per procedure, national trabecular stent utilization stably declined by 33%. Over the same period, goniotomy utilization increased by 61% and canaloplasty utilization increased by 19%, Total MIGS volume remained relatively stable. Industry stent-related TOV payments to ophthalmologists increased substantially in 2022 but subsequently declined in 2023. Stratified analyses demonstrated that reductions in trabecular stent utilization occurred across all levels of industry TOV payment recipients, including among physicians receiving the highest payment amounts. A large Medicare payment reduction for trabecular meshwork bypass stent implantation was associated with rapid declines in stent utilization and increased performance of alternative MIGS procedures. Concurrent increases in industry TOV payments did not offset these shifts, suggesting that Medicare payment policy exerted a stronger influence on procedural selection than industry financial incentives. These findings highlight the sensitivity of surgical practice patterns to payment policy and suggest that even substantial industry payments may have limited influence when insurer payments change dramatically.