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 FAK and the Wnt/β-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 proliferator- activated receptor-γ coactivator-1α (PGC-1α), and carnitine palmitoyl transferase-1α (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 SMG- inhibited 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.
Hepatic diseases are a serious concern across the globe. The death of liver cells is a significant cause of liver diseases. Phytochemicals fromAndrographis 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 comprisingA. paniculata and T. purpurea extracts can be a promising approach in treating hepatotoxicity.
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.
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 µg QiE/mg DM). In stem bark, alkaloids were also abundant in the hexane fraction (793.2 ± 25.32 µg QiE/mg DM). Polyphenols were most concentrated in the ethyl acetate fractions (209.64 ± 3.91 and 212.51 ± 1.29 µg 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 µg/mL). The ethyl acetate fractions of leaves and hexane fractions of both plant parts demonstrated moderate activity (IC50: 30.85–38.69 µg/mL). Against Pf Dd2, only the dichloromethane fractions and the stem bark ethyl acetate fraction showed moderate activity (IC50: 25.91–39.17 µg/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.
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 μg/mL vs. 4.971 μg/mL for free CTD) and induced S-phase 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.
Extracellular vesicles (EVs) research has gained a significant amount of attention in recent years. EVs are a heterogeneous group of different vesicles that vary in their origin, size, and function. The very nature of EVs, from their biogenesis to the contents of their cargo, offers many options for exploration. Despite being studied for a few decades now, there has not been an established standardized approach to isolation. However, the future use of EVs in clinical practice is conditioned by the optimization and standardization of isolation methods. In this study, we systematically compared common EV isolation techniques such as ultracentrifugation, precipitation, tangential flow filtration, and affinity-based methods using conditioned cell culture media as the source. Isolated EVs were analyzed using mass spectrometry to characterize their proteomic profiles, and the shared protein content was evaluated across datasets. The gene ontology enrichment was further assessed using three bioinformatics platforms. Among the tested methods, ultracentrifugation emerged as the most effective isolation method in our analysis, thus underscoring its importance among commonly used techniques. Despite its analytical robustness, this method is unsuitable for routine clinical workflows due to its complexity and time demands. Our findings highlight the need for a standardized, less strenuous EV isolation method for clinical applications.