Search Results (42,068)

Background/Objectives: RNA interference (RNAi) is a promising strategy for mitigating diseases at the molecular level. However, RNAi is limited by its instability in biological fluids and impermeability to cellular membranes. In response, our lab has previously patented a non-ionizable lipid nanoparticle (LNP) platform (R8-PLP) for RNAi therapeutic delivery. This formulation incorporates 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DSPE-PEG) to improve particle stability and drug retention. However, long-anchored PEGylated lipids like DSPE-PEG may impair internalization and stimulate immune responses. The literature suggests substituting short-anchored PEGylated-lipids like 1,2-dimyristoyl-rac-glycero-3-[methoxy(polyethylene glycol)-2000] (DMG-PEG) to attenuate these effects. Here, we evaluated whether substituting DMG-PEG for DSPE-PEG in our R8-PLP would improve in vitro cellular delivery and gene transfection without compromising in vitro critical quality attributes (CQAs) or increasing cytotoxicity. Methods: CQAs [encapsulation efficiency (EE%), particle size (nm), homogeneity (polydispersity index; PDI), and membrane zeta-potential] were assessed at assembly and after storage for up to 28 days at 4 °C. Additionally, in-serum stability at 4 °C and serum release kinetics at 37 °C were assessed. Human aortic smooth muscle cells (HASMCs) were treated with R8-PLPs and analyzed for cellular uptake (fluorometry), cytotoxicity (LIVE/DEAD stain), and gene modulation (qPCR). Results: DMG-PEG incorporation at variable mol% did not alter R8-PLP size, homogeneity, or siRNA EE% at assembly or after long-term storage, but did accelerate siRNA release kinetic profiles compared to DSPE-PEG controls. DMG-PEG substitution enhanced cellular uptake compared to DSPE-PEG R8-PLPs without increasing cytotoxicity. DMG-PEG incorporation also achieved significant silencing versus non-treated controls but did not improve gene silencing compared to DSPE-PEG R8-PLPs. Conclusions: Thus, DMG-PEG substitution did not enhance R8-PLP in vitro gene modulation efficacy despite improving cellular uptake and maintaining CQAs. Full article

Decarbonizing heavy-duty road freight transportation requires efficient energy management in zero-emission powertrains. This study investigates energy management strategies (EMSs) for a heavy-duty Fuel Cell Plug-in Hybrid Electric Vehicle (FC-PHEV). Rather than the typical charge-sustaining operation, these strategies are designed for charge-depleting operation, in which each route begins with a charged battery and ends at a lower state of charge (SOC), leveraging the vehicle’s plug-in capability. The EMSs are evaluated primarily in terms of energy consumption, while battery C-rate and fuel cell ramp rate are used as simple stress indicators for comparative analysis. A backward-facing vehicle model is developed to test several EMSs, including both optimization- and rule-based strategies. The Equivalent Consumption Minimization Strategy (ECMS) emerged as a promising option, motivating further testing with a forward-facing model and additional drive cycles. The simulation results show that ECMS consumed only 1.1% more energy than the global optimal solution found by Pontryagin’s Minimum Principle (PMP) and 7.5% less energy than a simple rule-based strategy, on average across five drive cycles. These results show that ECMS can be effective for a heavy-duty FC-PHEV operating in charge-depleting mode, extending its demonstrated applicability beyond charge-sustaining and light-duty vehicles. Full article

Objectives: This study aimed to investigate the biological role and molecular mechanism of the RNA m⁵C methyltransferase NSUN4 in cervical cancer progression, with a focus on its involvement in ferroptosis regulation. Methods: Differential expression and survival analyses were performed using TCGA and GEPIA datasets. Functional enrichment and GSEA identified pathways associated with NSUN4 dysregulation. NSUN4 expression was validated in clinical tissues by qRT-PCR, Western blot, and immunohistochemistry. Gain- and loss-of-function assays, including CCK-8, colony formation, and Transwell assays, were conducted to assess cell proliferation and invasion. Furthermore, a nude mouse subcutaneous xenograft model was established to validate the oncogenic role of NSUN4 in vivo. Ferroptosis was evaluated using specific inhibitors and measurement of GSH and ferroptosis-related proteins. RIP, m⁵C-RIP, RNA stability, and dual-luciferase assays were performed to explore the underlying mechanism. Results: NSUN4 was markedly upregulated in cervical cancer tissues and correlated with poor prognosis. Functionally, NSUN4 enhanced tumor cell growth, migration, and invasion while inhibiting ferroptosis. Mechanistically, NSUN4 bound to and stabilized C-MYC mRNA via m⁵C methylation, activating the PI3K/Akt signaling pathway and promoting ferroptosis resistance. Conclusions: NSUN4 promotes cervical cancer progression by stabilizing C-MYC mRNA through m⁵C modification, leading to PI3K/Akt activation and suppression of ferroptosis. These findings identify NSUN4 as a novel oncogenic regulator and potential therapeutic target in cervical cancer. Full article

Background: Recent studies strongly suggest that low intracranial pressure (ICP) may be involved in the pathogenesis of glaucomatous optic neuropathy. As retinal ganglion cells (RGCs) are highly susceptible to mitochondrial dysfunction, mitochondrial injury may be associated with optic neuropathy related to reduced ICP. In this study, aquaporin-1 (AQP1)-null mice were used to investigate whether reduced ICP is associated with alterations in mitochondrial structure and the release of optic atrophy type 1 (OPA1) and cytochrome c from mitochondria. Methods: Intraocular pressure (IOP) and ICP were measured in AQP1-null mice, and mitochondrial structural changes were examined using transmission electron microscopy (TEM). Total OPA1 and cytochrome c protein levels were evaluated using immunocytochemistry and Western blotting. Cytosolic and mitochondrial fractions were extracted from retinal tissues, and the subcellular distribution of OPA1 and cytochrome c was further analyzed by Western blotting. Bax and Bcl-2 expression levels were also detected. Results: TEM revealed mitochondrial fission, matrix swelling, and abnormal cristae depletion in the retinas of 1-, 3-, and 6-month-old AQP1-null mice. Morphometric quantification further confirmed significantly reduced mitochondrial length across all age groups and increased mitochondrial width at 1 and 6 months in AQP1-null mice compared with wild-type controls. Decreased retinal OPA1 immunoreactivity and protein expression were observed across all age groups of AQP1-null mice compared with age-matched C57BL/6 control mice. Subcellular fractionation showed increased mitochondrial release of OPA1 (at 3 and 6 months) and cytochrome c (at 1, 3, and 6 months) in the retinas of AQP1-null mice. Altered Bax expression was also detected in the retinas of AQP1-null mice with reduced ICP at all examined ages. Conclusions: Mitochondrial ultrastructural abnormalities, including fission and cristae depletion, altered OPA1 distribution, increased mitochondrial release of OPA1 and cytochrome c, and upregulated Bax expression were observed in the retinas of AQP1-null mice with reduced ICP. These concurrent changes indicate a close association between reduced ICP and retinal mitochondrial dysfunction. Maintaining mitochondrial integrity may therefore serve as a potential protective strategy against optic nerve degeneration in patients with chronic low ICP. Full article

Background: Hepatic ischemia–reperfusion injury (IRI) remains a major challenge in liver transplantation (LTx) and hepatectomy. Previous studies identified a 12/15-lipoxygenase (12/15-LOX)-driven lipid peroxidation cascade promoting cell death, whereas glutathione peroxidase 4 (GPx4)-dependent metabolism acts antagonistically. This study investigated whether tacrolimus protects against hepatic IRI through this redox axis. Methods: Male C57BL/6 mice underwent 65% partial hepatic warm ischemia and reperfusion with or without tacrolimus preconditioning. Liver tissue and serum were analyzed by spectral photometry, Western blotting, TUNEL assay, and serum enzyme measurement. Results were statistically analyzed and compared with previously published results of 12/15-LOX inhibition by baicalein pretreatment and its carrier DMSO. Also, the combination of both tacrolimus and baicalein was investigated. Results: Tacrolimus increased the oxidative glutathione activity quotient (GSSG/GSH) by 75.1% (p = 0.0302), attenuated MAPK signaling, reduced SAPK/JNK by 84.6% (p = 0.0059), with ERK1/2 showing a downward trend, decreased Caspase-3 activation by 66.9% (p < 0.001) and PARP cleavage by 59.9% (p = 0.0330), and lowered TUNEL-positive cell death by 61.8% (p = 0.0015). Tacrolimus achieved hepatoprotection comparable to 12/15-LOX inhibition, but without hepatotoxicity, whereas combined treatment conferred no additional benefit yet bore toxic properties. Conclusions: Tacrolimus preconditioning mitigates hepatic IRI through a glutathione-linked redox–signaling–cell death axis and exerts cytoprotective effects beyond immunosuppression. Full article

Background: Sézary syndrome (SS) is an aggressive leukemic variant of cutaneous T-cell lymphoma (CTCL) with distinct clinical and biological features compared to rarer entities such as primary cutaneous CD8⁺ aggressive epidermotropic cytotoxic T-cell lymphoma (PCAECTCL). Although recurrent genomic alterations in CTCL have been described, comparative analyses at the pathway level across biologically divergent subtypes remain limited. Here, we leveraged a conversational artificial intelligence (AI) platform for precision oncology to enable rapid, integrative, and hypothesis-driven interrogation of publicly available genomic datasets. Methods: We conducted a secondary analysis of somatic mutation and clinical data from the Columbia University CTCL cohort accessed via cBioPortal. Cases were stratified into SS (n = 26) and PCAECTCL (n = 13). High-confidence coding variants were curated and mapped to biologically relevant signaling pathways and functional gene categories implicated in CTCL pathogenesis. Pathway-level mutation frequencies were compared using Fisher’s exact tests, with effect sizes quantified as odds ratios. Tumor mutational burden (TMB) was compared using the Wilcoxon rank-sum test. Subtype-specific co-mutation patterns were evaluated using pairwise association analyses and visualized through oncoplots and network heatmaps. A conversational AI agent, AI-HOPE, was used to iteratively refine cohort definitions, prioritize pathway-level signals, and contextualize findings. Results: TMB was comparable between SS and PCAECTCL (p = 0.96), indicating no significant difference in global mutational load. In contrast, pathway-centric analyses revealed marked qualitative differences. SS demonstrated enrichment of alterations in epigenetic regulators, tumor suppressor and cell-cycle control pathways, NFAT signaling, and DNA damage response mechanisms, consistent with transcriptional dysregulation and immune modulation. PCAECTCL exhibited relatively higher frequencies of alterations involving epigenetic regulators and MAPK pathway signaling, suggesting distinct oncogenic dependencies. Co-mutation analysis revealed a more constrained and focused interaction landscape in SS, whereas PCAECTCL displayed broader and more heterogeneous co-mutation networks, indicative of divergent evolutionary trajectories. Notably, ERBB2 mutations were significantly enriched between subtypes (p = 0.031), highlighting a potential subtype-specific therapeutic vulnerability. Conclusions: This study demonstrates that SS is distinguished from PCAECTCL not by increased mutational burden but by distinct pathway-level architectures, particularly involving epigenetic regulation, immune signaling, and transcriptional control. These findings generate biologically grounded, testable hypotheses for subtype-specific therapeutic targeting and underscore the value of conversational AI as a scalable framework for accelerating discovery in translational cancer genomics. Full article

Background: This study investigated how chronic heat stress affects meat quality and post-slaughter muscle acidification in slow-growing yellow-feathered broilers, focusing on the roles of ALDOB and HSP90B1 in glycometabolism. Methods: From 100 to 120 days of age, broilers were kept either under thermoneutral conditions (25 ± 1 °C, N group) or cyclic heat stress (32 ± 1 °C for 9 h/day, H group). Meat quality traits (pH, shear force, drip loss, color) were measured at 0, 24, and 48 h of refrigeration (4 °C). Free amino acid and fatty acid profiles were analyzed. DF-1 cells were exposed to 43 °C for functional assays of ALDOB and HSP90B1. Results: Chronic heat stress reduced body weight, altered flavor precursors, and induced PSE-like characteristics (lower pH, higher shear force, increased drip loss, paler color), especially in leg muscles. ALDOB and HSP90B1 were upregulated in both tissues and cells. ALDOB overexpression promoted glucose consumption, while HSP90B1 suppressed lactic acid production. Conclusions: Chronic heat stress impairs growth and flavor precursors and exacerbates post-slaughter muscle acidification (primarily driven by ATP hydrolysis, with lactic acid as a secondary contributor). ALDOB and HSP90B1 may dually regulate glycometabolism under heat stress. Full article

Perineural spread (PNS) from cutaneous squamous cell carcinoma (cSCC) to the skull base is increasingly recognised as a route of cancer spread. Management historically involved definitive radiotherapy to treat PNS at the skull base. In the endemic region with modern magnetic resonance neurogram (MRN) and skull base surgical expertise, the outcome has improved over the years. With the advent of immunotherapy, the outcome may be maintained while preserving critical organs in the head and neck region. We conducted a critical review of the literature to establish the treatment outcomes and pattern of failure as practice evolved. Furthermore, we described our skull base surgical and radiotherapy management guideline and outlined the emerging paradigm shift in the immunotherapy era. Full article

Background: Trichosporon japonicum is a rare but highly lethal pathogen causing fungemia in immunocompromised patients. With the expanding use of chimeric antigen receptor T (CAR-T) cell therapy, the spectrum of opportunistic fungal infections is changing, yet data on T. japonicum infections in this setting remain scarce. Case Presentation: A 69-year-old man with diffuse large B-cell lymphoma developed catheter-associated fungemia after CAR-T cell reinfusion. He initially presented with neck pain and white oral mucosal patches, followed by fever four days later. T. japonicum was isolated from both peripheral blood and central venous catheter tip cultures, identified by microscopic examination, mass spectrometry, and molecular sequencing. Antifungal prophylaxis was initiated before fever onset based on close monitoring of white blood cell count, procalcitonin, interleukin-6, and C-reactive protein; treatment was subsequently adjusted according to species identification and antifungal susceptibility results. Infection was controlled within two weeks after catheter removal and immune recovery. The patient remained well at six-month follow-up. Conclusion: This case adds to the limited literature on T. japonicum fungemia in patients receiving CAR-T therapy. Our experience, together with a review of the literature, underscores that successful management requires prompt catheter removal, immune restoration, and combination therapy with voriconazole and amphotericin B, as echinocandin monotherapy should be avoided. Awareness of this pathogen in immunocompromised patients is critical. Full article

The cornea of the dromedary camel is essential for maintaining ocular clarity and protecting the eye in dry, dusty, and thermally stressful environments. Aquaporins are membrane channels that facilitate water transport, and AQP1 has been widely implicated in corneal fluid homeostasis in several species. The present work investigated, for the first time, the regional distribution of AQP1 in the camel cornea. Corneas collected from twelve healthy adult camels after slaughter were divided into nine anatomical regions: central (C), middle dorsal (MD), middle ventral (MV), middle nasal (MN), middle temporal (MT), peripheral dorsal (PD), peripheral ventral (PV), peripheral nasal (PN), and peripheral temporal (PT). Histological examination and immunohistochemistry were combined with digital morphometry to assess corneal layer thickness and AQP1 localization. AQP1 labeling was identified in the corneal epithelium, stromal keratocytes, and endothelium. Epithelial staining differed among regions and was most pronounced in the peripheral nasal region, whereas stromal keratocytes and endothelial cells showed strong and relatively uniform immunoreactivity. These findings indicate that AQP1 is broadly expressed in the camel cornea and likely contributes to regional control of hydration and tissue maintenance in an arid-adapted species. Full article

Background: Emergency department evaluation of suspected urolithiasis increasingly relies on non-contrast CT, yet not all patients require imaging. Existing clinical prediction rules help stratify stone probability, but by converting continuous measurements into fixed binary indicators, they offer little insight into why a particular patient is at risk or how much uncertainty remains after each testing stage—questions that bear directly on individualized diagnostic decisions. Methods: We retrospectively analyzed 1000 ED patients with suspected urolithiasis who underwent non-contrast CT (stone prevalence 85.0%). A gradient boosting classifier was trained on 17 continuous clinical and laboratory features and compared against binary-thresholded counterparts and an established scoring system; the 17-feature model achieved AUC 0.771 (95% CI 0.726–0.813) versus 0.723 (95% CI 0.675–0.771) for the reference score on this cohort (DeLong p = 0.001). Individual predictions were explained using an interventional Shapley value approach, and a Shannon entropy-based framework was applied to quantify the marginal diagnostic contribution of each sequential testing stage. Results: Held-out permutation importance identified red blood cell count on microscopy, age, pain duration, and prior stone history as the most influential predictors. Several features showed non-linear contributions that diverged from conventional binary thresholds: creatinine effect crossed zero near 0.90 mg/dL and pain duration peaked between 2 and 5 h. C-reactive protein, absent from existing scoring systems, emerged as a meaningful negative predictor. Sequential entropy analysis showed that dipstick urinalysis provided the largest marginal information gain among non-history stages (6.1% of prior entropy), while physical examination contributed 2.3%. A prevalence sensitivity analysis projected that the framework’s threshold behavior would differ substantially in lower-prevalence populations, underscoring that the cohort-specific cut-points are not portable decision rules. We therefore position the framework as a reasoning aid that complements clinical judgment and imaging, not as a stand-alone triage tool. Conclusions: Explainable machine learning can address questions that aggregate discrimination metrics cannot: which features drive risk for a given patient, how those effects behave across the continuous measurement range, and how much diagnostic uncertainty each testing stage resolves. The Shapley-based explanations and entropy framework developed here offer a structured approach to individualized diagnostic reasoning in the ED evaluation of suspected urolithiasis, functioning as an interpretive adjunct to, rather than a replacement for, existing clinical tools and CT imaging. Full article

Background: Prenylated chalcones are recognized for their beneficial nutritional properties and have attracted increasing interest due to their anticancer activities, which involve various mechanisms and pathways. In the current study, we investigated the influence of prenylated chalcone xanthohumol (XH) and its two minor derivatives xanthohumol C (XHC) and 1″,2″-dihydroxantohumol C (DHXHC) on the formation of reactive oxygen species (ROS), causing oxidative stress. Concomitantly, we studied the effect of mitochondrial transmembrane potential changes on human skin cancer, namely Merkel cell carcinoma (MCC13). Methods: The cancer cells were treated with the mentioned compounds for 24 and 48 h at various concentrations. Results: Our findings showed that ROS generation was dose-dependent at 24 h for xanthohumol, whereas for xanthohumol C and 1″2″-dihydroxanthohumol C, a significant increase in ROS occurred only at the highest concentration (100 μM) after 48 h. Mitochondrial membrane potential was significantly diminished by all the compounds. Conclusions: Taken together, our results indicate that the aforementioned chalcones exhibit cytotoxic activity against the MCC13 cell line and may be promising candidates for further investigation as anticancer agents. Full article

Hepatocellular carcinoma (HCC) is the most common type of liver cancer that is mostly preceded by cirrhosis, with a high mortality rate. Therefore, diagnosis is critical in the early stages. In this study, we explored the liver expression of metabotropic glutamate receptor 3 (mGluR3), a group II mGluR, during the progression from fibrosis to cirrhosis and, ultimately, to HCC induced by diethylnitrosamine (DEN) in rats. We found that mRNA expression of mGluR3 (Grm3) was upregulated in HCC, while the protein level was significantly increased from the cirrhosis stage, and even more in HCC. Grm3 correlated with interleukin-6 (Il6) and transforming growth factor-β (Tgfb) mRNA expression. Furthermore, serum and intrahepatic glutamate concentrations were augmented in HCC. Immunohistochemical analysis revealed that mGluR3 is expressed in hepatocytes and non-parenchymal cells (endothelial cells and macrophages), and we observed a positive signal in the cytoplasmic membrane, cytoplasm, and nuclei of tumor and non-tumor cells. We confirmed that normal hepatocytes (C9 cell line) express low levels of mGluR3 protein and HCC-derived cells (HepG2) express high levels of this receptor. Using HepG2 cells, we observed that mGluR3 activation by glutamate and the group II-selective agonist LY354740 treatments were functional, as both inhibited cAMP generation induced by forskolin and increased cellular viability with no effect on dead cells. These results showed that mGluR3 is differentially expressed throughout the progression of liver pathologies, is associated with the inflammatory environment, and plays a role in HCC cell survival, with potential utility as an early biomarker and therapeutic target. Full article

This paper investigates the safety behavior of degraded lithium-ion battery modules taken from a 2016 Nissan Leaf (30 kWh, 106,394 km). The vehicle exhibited typical failure symptoms, including P33E6 faults, sudden range drops, and activation of turtle mode under load. Initial diagnostics based on LeafSpy data revealed strong cell imbalance, with a voltage spread exceeding 2.3 V under high current (≈170 A). The weakest cells dropped close to 1 V, suggesting severe internal degradation. To better understand this behavior, selected modules (cells 73–88) were removed and tested under controlled laboratory conditions. Capacity measurements in a 16S2P configuration showed 49.8 Ah in the 4.1–3.1 V range, corresponding to a state of health of about 59%, which is consistent with BMS estimates. However, high-current discharge tests on the weakest segment revealed a much more critical picture. One cell experienced rapid voltage collapse (from ~4.0 V to ~1.2 V), accompanied by a sharp increase in voltage divergence and visible thermal effects. Infrared observations indicated localized heating up to 43 °C and irreversible swelling, pointing to early-stage electro-thermal instability. These results suggest that moderate SOH values do not necessarily reflect actual safety margins under dynamic load conditions. Overall, the study shows that simple OBD-based diagnostics can help identify problematic modules, but additional load testing is necessary to assess real safety risks in aged EV battery systems. Full article

Since MDCK cells are inherently tumorigenic, their safety in vaccine production has long been a concern; thus, establishing a screening method for low-tumorigenic cells is of great significance for influenza vaccine development. This study successfully obtained a low-tumorigenic MDCK cell line through monoclonal screening and systematically evaluated its potential as a cellular substrate for influenza vaccines using male nude mice (BALB/c nu/nu, 4–7 weeks old) for tumorigenicity assessment. Comprehensive analysis of the biological characteristics of the screened cells—including growth curves and transcriptomic features—showed that the cell line exhibits stable growth and consistent traits. Transcriptomic comparison was performed between two defined biological states: parental MDCK cells (SQ group) and the low-tumorigenic clone MDCK-20B9 (SH group). Transcriptomic analysis revealed good dispersion among samples and an overall consistent gene expression distribution. Differential expression analysis identified a total of 2198 differentially expressed genes, including 902 upregulated and 1296 downregulated genes. GO functional enrichment analysis indicated that these genes are mainly involved in biological processes such as acute-phase response, retinol metabolism, mitotic chromosome condensation, and cell migration; are enriched in cellular components such as kinetochores and the extracellular matrix; and are associated with molecular functions including calcium ion binding and the Wnt signaling pathway. KEGG pathway analysis further revealed that the differentially expressed genes are significantly enriched in key pathways such as cancer pathways, cell cycle, and cell adhesion molecules. The expression trends of five key differentially expressed genes were validated by RT-qPCR. In summary, this study successfully screened a stable and consistent low-tumorigenic MDCK cell line, providing a theoretical basis and practical foundation for its use as a cellular substrate in influenza vaccine development. Full article