The fullerene C60 is widely studied as a potential nanoscale drug carrier for hydrophobic anticancer compounds; however, its practical application in biomedicine has been limited due to low polarity and water solubility. Herein, we utilized Density Functional Theory (DFT) calculations to study the impact of endohedral encapsulation of several heteroatoms (boron, nitrogen, silicon, and phosphorus) on the structure and electronic properties of C60. Each atom was placed within the confines of the fullerene cage adjacent to the same pentagonal carbon group to compare interactions and stability. Optimization results showed minimal impact on C-C bond length due to encapsulation, maintaining a stable fullerene cage structure. Calculated interaction energies showed that encapsulation of B, N, and Si within the fullerene cage is energetically favourable, whereas P encapsulation is less favourable. C60N exhibited the highest interaction as a result of large charge transfer between the cage and nitrogen atom, while C60Si was found to have the greatest dipole moment, suggesting increased polarity. HOMO-LUMO analysis showed a decreased gap upon encapsulation of all molecules, which suggests higher chemical softness. Density of states (DOS) analysis revealed noticeable changes in the electronic structure following encapsulation. The suitability of the investigated systems as drug carriers was evaluated using temozolomide (TMZ) as a model anticancer drug. Among the studied complexes, SiC60-TMZ exhibited favourable interaction characteristics and enhanced solvation stability. Overall, C60Si presents an optimal balance of stability, polarity, and electronic softness, highlighting its potential as an efficient nanocarrier for hydrophobic drug delivery applications.
Choline kinase alpha (CHKA) plays an important role in phospholipid metabolism and is frequently overexpressed in several cancers, where it contributes to tumor growth and survival. In this study, we investigated whether microRNA-32-5p (miR-32-5p) regulates CHKA expression and influences malignant phenotypes in human breast cancer cells. Bioinformatic prediction and luciferase reporter assays suggested a potential interaction between miR-32-5p and the CHKA 3' UTR. Transfection of miR-32-5p into MCF7 cells significantly reduced CHKA mRNA and CHKA protein expression. This downregulation was associated with increased apoptosis, G0/G1 cell-cycle arrest, and reduced migration of MCF7 cells. Reduced phosphorylation of ERK and mTOR was also observed, suggesting decreased activation of MAPK/mTOR signaling pathways. In contrast, although CHKA expression was reduced in non-tumorigenic MCF10A cells, apoptosis was not induced. These findings indicate that miR-32-5p regulates CHKA expression and suppresses malignant phenotypes in breast cancer cells, highlighting its potential role in modulating CHKA-associated signaling pathways.
This study evaluated the effects of an alkaloid-rich leaf extract of Dalbergiella welwitschii (Baker) Baker f. on hepatic carbohydrate metabolism in streptozotocin-induced diabetic rats. Diabetes was induced in rats, which were then treated with low (50 mg/kg) and high (100 mg/kg) doses of the extract, or metformin (200 mg/kg), for 20 days. Key hepatic enzymes-glycogen phosphorylase, glucose-6-phosphatase, lactate dehydrogenase, fructose-1,6-bisphosphatase, glycogen synthase, and hexokinase-along with hepatic glycogen and insulin levels were measured. Treatment with the extract significantly restored enzyme activities, increased insulin levels, and enhanced glycogen content. These results indicate that D. welwitschii extract can modulate hepatic carbohydrate metabolism and may have potential therapeutic benefits for diabetes management.
In this study, we focus on the enzyme-assisted extraction (EAE) of bioactive compounds from Posidonia oceanica, a Mediterranean seagrass of ecological importance that is often treated as waste. Increasing evidence highlights its beneficial properties, making it a promising source of compounds for the production of value-added compounds. A Taguchi experimental design was employed to optimize the EAE, using the cellulolytic commercial enzyme preparation Cellic CTec3 (R) HS. The effects of key parameters, namely extraction time, solid-to-liquid ratio, and enzyme loading, were evaluated for total phenolic content (TPC) and total flavonoid content (TFC). Optimization was achieved using "the larger-the-better" approach, and under optimal conditions (solid-to-liquid ratio 1% w/v, enzyme loading 200 U/g, and extraction time 6 h), TPC and TFC reached 27.54 +/- 0.84 mg GAE/g DW and 9.22 +/- 0.84 mg CAE/g DW, respectively, representing approximately a ten-fold and more than five-fold increase compared to conventional ethanol:water extraction. An additional hemicellulolytic enzyme preparation, Viscoferm (R) was evaluated under the optimized conditions, and EAE with Cellic CTec3 (R) HS yielded higher TPC and TFC. The optimized extracts obtained with Cellic CTec3 (R) HS and Viscoferm (R) were further assessed for their antioxidant activity, anti-aging activity (inhibition of tyrosinase activity), anti-diabetic activity (inhibition of amylase and alpha glucosidase activity), and antibacterial activity (inhibition of E. coli growth). The Cellic CTec3 (R) HS extract yielded highest antioxidant activity (IC50 =203.72 +/- 23.99 mu L extract/mL) and antibacterial activity (21.37 +/- 2.00 %), whereas the Viscoferm (R) extracts demonstrated stronger antidiabetic activity
Objective: Hepatocellular carcinoma (HCC) remains a molecularly complex malignancy with limited therapeutic options. Cantharidin (CTD), a natural terpenoid, exhibits potent antitumor activity but suffers from poor bioavailability and dose-limiting toxicity. This study aimed to develop CTD-loaded liposomes (CTD-Lips) and investigate their molecular mechanisms in enhancing apoptosis and cell cycle regulation in HCC. Methods: CTD-Lips were prepared using an optimized thin-film method and thoroughly characterized for size, encapsulation efficiency, and drug release profile. Cellular uptake was evaluated using coumarin-6-loaded nanoparticles in HepG2 cells. Cytotoxicity was assessed by CCK-8 assay, while apoptosis and cell cycle distribution were analyzed via flow cytometry. In vivo antitumor efficacy and systemic safety were evaluated in HepG2-xenografted nude mice compared to cisplatin, with comprehensive histopathological and biochemical analyses. Results: Optimized CTD-Lips showed nanoscale characteristics (120.80 nm) with high encapsulation efficiency (96.20%) and sustained release. Cellular uptake was significantly enhanced, leading to superior cytotoxicity (IC50: 1.415 mu g/mL vs. 4.971 mu g/mL for free CTD) and induced Sphase cell cycle arrest. Molecular analysis revealed that CTD-Lips promoted apoptosis through enhanced DNA damage response. In vivo, CTD-Lips achieved 62.4% tumor growth inhibition, outperforming cisplatin, while maintaining systemic safety with no significant hematological or biochemical alterations. Conclusions: CTD-Lips represent a molecularly targeted nanoplatform that enhances CTD's antitumor efficacy through regulation of cell cycle progression and apoptosis pathways. This study provides molecular-level insights into CTD-Lip-mediated anticancer mechanisms, supporting its potential as a novel molecular-targeted therapy for HCC.
Malaria is a potentially fatal disease caused by Plasmodium parasites transmitted by infected Anopheles mosquitoes, and its management is increasingly hampered by growing drug resistance. This study aims to evaluate the antiplasmodial activity of different fractions obtained from the leaves and stem bark of Spathodea campanulata. Dried plant materials were extracted to produce dichloromethane, ethyl acetate, and hexane fractions. Phytochemical screening and absorption spectrophotometry were used to identify and quantify secondary metabolites. Antiplasmodial activity was assessed against Plasmodiun falciparum 3D7 and Dd2 strains, and IC50 values were determined. Leaves showed high alkaloid levels, especially in the hexane, ethyl acetate, and dichloromethane fractions (1028.36 +/- 21.36, 950.03 +/- 25.44, and 641.8 +/- 8.16 & micro;g QiE/mg DM). In stem bark, alkaloids were also abundant in the hexane fraction (793.2 +/- 25.32 & micro;g QiE/mg DM). Polyphenols were most concentrated in the ethyl acetate fractions (209.64 +/- 3.91 and 212.51 +/- 1.29 & micro;g GaE/mg DM for leaves and stem bark), and flavonoid levels were highest in the ethyl acetate and hexane fractions of both plant parts. Dichloromethane fractions of leaves and stem bark, as well as the ethyl acetate stem bark fraction, showed good activity against Pf 3D7 (IC50: 16.69-19.84 & micro;g/mL). The ethyl acetate fractions of leaves and hexane fractions of both plant parts demonstrated moderate activity (IC50: 30.85-38.69 & micro;g/mL). Against Pf Dd2, only the dichloromethane fractions and the stem bark ethyl acetate fraction showed moderate activity (IC50: 25.91-39.17 & micro;g/mL). These activities are likely linked to alkaloid, polyphenol, and flavonoid content. The most active fraction was the dichloromethane fraction of the stem bark, which was effective on both strains and may represent potential therapeutic alternatives.
Glaucoma, a leading cause of irreversible blindness, poses significant diagnostic challenges due to its subtle onset and progressive nature. Current methods using color fundus images often fail to detect early-stage glaucomatous changes. To address this, we introduce Glauco-Net, a novel algorithmic framework integrating four transformative innovations. First, the spectral-adaptive retinal reconstruction engine (SARRE) reconstructs hyperspectral details from RGB fundus images, uncovering latent spectral biomarkers of early glaucomatous damage. Second, the vascular topology signature embedding (VTSE) employs graph neural networks to analyze retinal vascular topology, capturing structural irregularities linked to disease progression. Third, the temporal texture evolution transformer (TTET) models spatiotemporal texture dynamics in sequential fundus images, detecting subtle textural shifts indicative of glaucoma. Finally, the peripapillary light scattering dynamics profiler (PLSDP) simulates light-tissue interactions to identify peripapillary structural anomalies linked to glaucomatous cupping and atrophy. These innovations are fused into a unified deep learning pipeline, achieving SOTA performance in glaucoma diagnosis. Evaluated on large-scale clinical datasets, Glauco-Net achieved an AUC exceeding 0.97, demonstrating superior sensitivity and specificity compared to existing methods. By leveraging advanced spectral, topological, and dynamic analyses, this framework not only enhances early detection but also provides deeper insights into glaucoma pathogenesis. Our work represents a paradigm shift in computational ophthalmology, offering a robust, multimodal approach to glaucoma diagnosis and paving the way for personalized monitoring and intervention strategies. The source code of the proposed Glauco-Net is available at https://github.com/livingjesus/Glauco-Net.
Hepatic diseases are a serious concern across the globe. The death of liver cells is a significant cause of liver diseases. Phytochemicals from Andrographis paniculata and Tephrosia purpurea are reported to have hepatoprotective potential. The oral delivery of extracts containing phytochemicals using a tablet can produce a synergistic effect and enhance safety and patient compliance. This study aimed to extract phytochemicals from those plants and investigate their phytochemical potential, the pre-and post-compression attributes of the extracts' powder, their compression into tablets, and the in vivo performance of the tablets in paracetamol-induced hepatotoxicity in Wistar rats. Phytochemical screening of two extracts revealed the presence of flavonoids, phenols, carbohydrates, terpenoids, tannins, proteins, cholesterol, diterpenes, and glycosides. Evaluation of pre-compression properties revealed good flowability and suitability for compression into tablets. The post-compression quality control parameters were found to match the standard. The polyherbal tablet exhibited dose-dependent hepatoprotective activity in rats. The histological study revealed that the administration of 650 mg/kg body weight of paracetamol resulted in damage to the liver, as evidenced by increased levels of various enzymes, such as serum glutamic-oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), and alkaline phosphatase (ALP), total cholesterol (TC), triglycerides (TG), and total proteins (TP). Liver enzyme levels were significantly (p < 0.005) decreased after treatment with the polyherbal tablet formulations. Furthermore, histological analysis showed that the liver anatomy of the rats given polyherbal tablets was restored to near-normal architecture. The polyherbal tablet comprising A. paniculata and T. purpurea extracts can be a promising approach in treating hepatotoxicity.
Background: Aerospace microgravity (AMG) poses a major threat during spaceflight, impairing osteoblast differentiation (OBD) and causing bone loss. To replicate AMG conditions for ground-based experimental procedures, simulated microgravity (SMG) has been implemented to explore the molecular mechanisms of AMG-induced alterations in osteoblast differentiation. However, SMG's metabolic implications contributing to defective osteoblast differentiation remain unexplored. Methods: To investigate this, we investigated effects of SMG on pre-osteoblast MC3T3-E1 cells using Western-blotting to analyze expression of metabolic regulators and Seahorse assays to characterize cellular metabolism. The cytotoxic necrotizing factor-1 (CNF1), an activator of focal adhesion kinase (FAK), was applied to assess its ability to modulate SMG-inhibited OBD. Results: Our results showed that, besides its established inhibition of FAX and the Wnt/beta-catenin signaling cascade, SMG also induced a metabolic shift from fatty acid oxidation (FAO) to glycolysis by decreasing mitochondrial content and reducing expression of metabolic regulators [sirtuin-1 (SIRT1), peroxisome proliferatoractivated receptor-gamma coactivator-1 alpha (PGC-1 alpha), and carnitine palmitoyl transferase-1 alpha (CPT1A)] that are critical for mitochondrial biogenesis and FAO capacity. CNF1 exposure was found to counteract the SMG's inhibitory influence by upregulating expression of above metabolic regulators and restoring mitochondrial content and FAO as the primary cellular metabolism and thereby rescuing OBD. Altogether, our findings emphasize that FAK activation plays a critical role in restoration of SMGinhibited osteoblast differentiation by triggering transcriptional Wnt/B-Catenin-BMP2 (bone morphogenic protein-2)-COL1 (type-1 collagen) and metabolic SIRT1-PGC1a-CPT1A pathways. Conclusion: Our data shed light on FAK as a potential therapeutic target to mitigate SMG-driven bone-loss for astronauts and attenuate bone defectiveness for clinical osteoporosis.
MicroRNAs (miRNAs) are key post-transcriptional regulators of gene expression and have been associated with pathological processes, including oncogenesis. In this study, the interaction between miRNAs and phosphoinositides, particularly phosphatidylinositol 4,5-bisphosphate (PIP2), was explored by pull-down assays, followed by RNA sequencing (RNA-seq) analysis in HeLa cells. Four miRNAs with specific affinity toward PIP2 and PI4P were identified, including hsa-miR-3181-1, hsa-miR-4720, hsa-miR-142, and hsa-miR-940. Structural analysis suggested that the interaction with lipids is mediated by secondary conformations rather than by sequence homology. Furthermore, two of these miRNAs were present in human blood fractions, with particular emphasis on hsa-miR-142, whose plasma distribution suggests a potential role as a phosphoinositide-mediated long-range signaling molecule. These miRNAs have been associated with tumor-related processes, suggesting a potential, although not yet confirmed, relevance as noninvasive biomarkers or therapeutic targets in cancer.
Although cardiovascular diseases, especially diabetic cardiomyopathy (DCM), account for up to 80% of deaths in diabetic patients, the pathophysiological changes driven by diabetes on cardiovascular function are poorly defined and treated. As with many medical conditions, increased levels of Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and Signal transducer and activator of transcription (STAT)3 are significant aspects of DCM pathophysiology. Research indicates that NF-κB dimer composition and nuclear (canonical) and mitochondrial (noncanonical) STAT3 interact across human cells to regulate the melatonergic pathway. Given the beneficial effects of melatonin across almost all cardiovascular diseases, including DCM, it is amiss that the melatonergic pathway has not been investigated in cardiomyocytes, cardiac fibroblast, or endothelial cells. In this article, we reviewed DCM pathophysiological factors/processes and link these to wider bodies of data on the regulation of the melatonergic pathway, providing a framework that better integrates previous disparate bodies of data on DCM, while indicating clear future research and treatment implications.
Cancer has emerged as a significant global health challenge, with both its occurrence and death toll rising annually. Studies have shown that early, precise diagnosis and targeted cancer therapy can effectively reduce the mortality rate of malignant tumors. As an emerging field, nanomedicine seeks to unify cancer diagnosis and therapy within a single platform, enabling early detection, precise drug targeting, and minimized harm to healthy tissues. In recent years, nanotechnology has emerged as a powerful tool in oncological applications, revolutionizing both cancer detection and therapeutic interventions, and clinical treatment strategies have also shown a significant trend of shifting from single therapy to combination therapy. Research demonstrates that multimodal combination therapy not only combines the benefits of individual treatment modalities but also generates synergistic effects, yielding significantly superior clinical outcomes compared to monotherapies or simple treatment combinations. Consequently, the integration of nanotechnology with multimodal synergistic approaches has emerged as an innovative paradigm in oncological therapeutics. This article provides a systematic review of the unique characteristics and functional mechanisms of various nanomaterials, while examining their translational applications in oncological diagnostics and therapeutics through nanomedicine approaches.
Chimeric Antigen Receptor T-cell (CAR-T) therapy has emerged as one effective treatment against complex diseases, including malignancies, infectious disorders, and autoimmune conditions. However, its clinical applications remain hindered by severe side effects, including cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity syndrome (ICANS). Recent innovations in genome editing and computational biology, specifically CRISPR/Cas9 and artificial intelligence (AI), offer promising avenues to improve CAR-T safety and efficacy. In this article, we reviewed recent advances in CAR-T therapy, highlighting the key role CRISPR/Cas9 plays in various aspects of CAR-T therapy through gene editing. We then studied and analyzed how AI leverages large datasets and its powerful learning capabilities to advance CAR-T therapies and CRISPR/Cas9. Finally, focusing on CAR-T therapy, we discussed how CRISPR/Cas9 and AI work synergistically to advance the development of CAR-T therapy, including reducing toxic side effects, improving therapeutic efficacy, sustaining long-term outcomes, discovering new therapeutic targets, and ensuring safety monitoring. By combining the predictive power of AI with the precision of CRISPR/Cas9, researchers can develop next-generation immunotherapies that are safe, effective, and tailored to the patients' requirements. This synergy can address previously untreatable diseases and reshape the landscape of precision medicine.
Our aim of this work was the development of positively charged D-alpha-tocopheryl glycol succinate 1000 (TPGS)-chitosan nanocapsules (TCNs) as a non-viral vector for effective genome editing. A two-step process was used to create cationic TCNs. TPGS was initially esterified using succinic anhydride to add a carboxylic acid group. Then, using a complex coacervation technique, ionic cross-linking was used to join the activated TPGS with chitosan. Fourier transform infrared (FTIR) spectroscopy was used to validate the synthesis of TCNs. A gel retardation experiment was performed to evaluate TCNs/pDNA complex entrapments at various N/P ratios (2:1, 5:1, and 10:1). The physicochemical properties of the TCNs/pDNA complex were identified. Plasmid DNA was protected from DNase I activity through TCN complexation. HEK293 and GTM3 cell lines were used to test in vitro transfection efficiency. At N/P 5:1, TCNs/pDNA exhibited the maximum transfection efficiency, which was comparable to Lipofectamine 3000TM. Cellular uptake and localization studies confirmed effective delivery of Cas9-GFP plasmids using TCNs. A T7 endonuclease mismatch detection assay further demonstrated effective targeting and editing of the MYOC gene in HEK293 cells using our TCN delivery system. Based on these findings, cationic TCNs represent a promising non-viral gene delivery platform for potential treatment of myocilin-associated glaucoma.
Cyanobacteria constitute a group of photooxybacteria. It is difficult to provide an estimate of their diversity due to the ongoing debate on the usefulness of various species concepts and methodologies used to delineate them. They occupy, thrive, and proliferate in diverse habitats, exhibiting unparalleled physiological plasticity and adaptations, potentiated by diverse mechanisms and the production of a range of novel molecules. Human use of cyanobacteria (and also other microalgae) and products thereof is not new, with reports from before 2700 BC. In recent times, some ecologically relevant molecules, such as cyanotoxins to deter grazers and osmoprotectants for survival in saline environments, have found multiple human uses, from feeding products to therapeutics and energy. Technological advancements have allowed precise identification and a better mechanistic understanding of their production on a commercial scale. Here, we discussed some of these commercially useful molecules and their production mechanisms, scaling-up possibilities, cost-effectiveness, and challenges and opportunities associated with future research.
