Search Results (6,473)

Continued population growth demands a significant increase in agricultural production to ensure food security. However, agricultural output is limited by environmental crises and the negative impacts of open-field farm practices. As an alternative, vertical farming techniques, such as aeroponics, can be utilized to optimize the use of resources. However, the uneven size and distribution of spray droplets in aeroponics, issues that affect root development and nutrient delivery, continue to be problematic in spray performance analysis. In aeroponics, nutrient solutions are delivered to plant roots through pressurized nozzles, and the effectiveness of this delivery depends on the spray characteristics. Variations in flow rates directly affect droplet size, density, and coverage, which in turn influence nutrient uptake and crop growth. In this study, the flow rate was adjusted (3, 4.5, and 6 L/min) to quantitatively analyze spray performance using water-sensitive paper (WSP) as a deposit collector via a quick assessment method. Subsequently, image-processing techniques such as threshold segmentation and morphological operations were applied to isolate individual spray droplets on the WSP images. This technique enabled the quantification of the droplet’s coverage area, size, density, and uniformity to effectively evaluate spray performance. One-way ANOVA indicated that all the spray parameters varied significantly with respect to the flow rate (p < 0.05): For example, the average diameters of the droplets increased from 0.73 mm at 3 L/min to 1.29 mm at 6 L/min. The droplets’ densities decreased from 85.53 drops/cm² to 30.00 drops/cm² across the same flow range. The average uniformity index improved from 30.53 to 15.95 as the flow rate increased. These results indicate that the application of WSP is an effective and scalable approach for analyzing spray performance in aeroponics, as WSP can be rapidly digitized with simple tools, such as a cell phone camera, avoiding the limitations of flatbed scanners or specialized imaging systems. Full article

TRIC-A is an intracellular cation channel enriched in excitable tissues that is recently identified as a key modulator of sarcoplasmic reticulum (SR) Ca²⁺ homeostasis through direct interaction with type 2 ryanodine receptors (RyR₂). Given the intimate anatomical and functional coupling between the SR and mitochondria, we investigated whether TRIC-A contributes to SR–mitochondrial crosstalk under cardiac stress conditions. Using a transverse aortic constriction (TAC) model, we found that TRIC-A^−/− mice developed more severe cardiac hypertrophy, underwent maladaptive remodeling, and activated apoptotic pathways compared with wild-type littermates. At the cellular level, TRIC-A-deficient cardiomyocytes were more susceptible to H₂O₂-induced mitochondrial injury and displayed abnormal mitochondrial morphology. Live-cell imaging revealed exaggerated mitochondrial Ca²⁺ uptake during caffeine stimulation and increased propensity for store-overload-induced Ca²⁺ release (SOICR). Complementary studies in HEK293 cells expressing RyR₂ demonstrated that exogenous TRIC-A expression attenuates RyR₂-mediated mitochondrial Ca²⁺ overload, preserves respiratory function, and suppresses superoxide generation. Together, these findings identify TRIC-A as a critical regulator of SR–mitochondrial Ca²⁺ signaling. By constraining mitochondrial Ca²⁺ influx and limiting oxidative stress, TRIC-A safeguards cardiomyocytes against SOICR-driven injury and confers protection against pressure overload-induced cardiac dysfunction. Full article

Mutant yeast strains with altered sensitivity to heavy metals are crucial for revealing the mechanisms of metal absorption and detoxification, as well as for bioremediation of these pollutants. Here, we show that a knockout of the PHO87 gene encoding the low-affinity phosphate transporter of the cytoplasmic membrane of S. cerevisiae increased resistance to manganese, silver, and vanadate ions. However, a knockout of PHO90 (PHO87 paralog) did not affect the sensitivity to silver and vanadate ions but increased sensitivity to manganese ions. The Δpho87 cells accumulated 10 times less manganese compared to the wild-type cells, while the Δpho90 cells accumulated two times more manganese compared to the wild-type cells, when grown in YPD with 2 mM MnSO₄. The polyphosphate content of the Δpho84, Δpho87, and Δpho90 cells cultivated at high phosphate concentration did not differ from that of the wild-type strain. In the presence of 2 mM MnSO₄, Δpho87 cells contained several times less polyphosphates, and Δpho90 cells contained more short-chain polyphosphates than the cells of the wild-type strain. We hypothesize that phosphate carriers participate in the regulation of heavy metal uptake, and the respective knockouts are useful in bioremediation and bioassay of these pollutants. Full article

An integrative approach combining network pharmacology, molecular docking, and cellular assays was used to elucidate the potential mechanisms by which the n-butanol extract of Biebersteinia heterostemon ameliorates type 2 diabetes mellitus (T2DM). Chemical constituents of the n-butanol extract were identified via ultra-high-performance liquid chromatography coupled with Q-Exactive Orbitrap mass spectrometry. Active compounds and T2DM-related targets were retrieved from public databases, and intersecting targets were identified. Protein–protein interaction (PPI) networks were constructed using the STRING database, while Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed via the DAVID database. A comprehensive “drug–compound–target–disease–pathway” network was established, and molecular docking was conducted to evaluate binding affinities of key compounds to core targets. Functional validation was performed in insulin-resistant cell models. Network pharmacology analysis identified 37 active constituents within the extract and 222 overlapping targets associated with T2DM. GO enrichment indicated involvement in protein phosphorylation, MAPK cascade activation, and negative regulation of apoptosis. Key signaling pathways included PI3K/AKT and lipid and atherosclerosis pathways. Molecular docking revealed strong binding affinities (binding energies ≤ −9.3 kcal·mol⁻¹) between core compounds—such as cheilanthifoline, glabridin, acetylcorynoline, skullcapflavone II, liquiritigenin, and dinatin—and pivotal targets including GAPDH, AKT1, TNF, SRC, EGFR, and PPARγ. In vitro experiments demonstrated that the extract significantly enhanced glucose uptake and glycogen synthesis in insulin-resistant cells, while suppressing oxidative stress and the expression of pro-inflammatory mediators such as TNF-α, MMP9, and IL-6. Collectively, B. heterostemon shows potential as an effective intervention for T2DM by targeting key molecular pathways, improving insulin sensitivity, and mitigating oxidative stress and inflammation in insulin-resistant cells. Full article

Most clinically used antibiotics exert their effects by targeting essential intracellular components of bacterial cells. Therefore, enhancing their ability to traverse the bacterial envelope is crucial for restoring or improving therapeutic efficacy. We investigated the potential of outer membrane (OM)-disrupting agents—EDTA, NV716, colistin, and squalamine—to potentiate antibiotic activity against the multi-drug-resistant pathogen Pseudomonas aeruginosa. Our objective was to assess the therapeutic value of this strategy while also delineating its limitations by comparing responses across antibiotic classes with diverse chemical structures and pharmacological profiles. Beyond lipophilicity, we analyzed three additional physicochemical descriptors likely to influence OM permeability: molecular surface area, polarizability, and polar surface area. Our findings offer practical insights for the rational design of antibiotic–adjuvant combinations. While each descriptor provides valuable interpretive information, none alone reliably predicts OM-mediated potentiation. Instead, these factors should be viewed collectively within a multidimensional physicochemical profile, where optimal ranges of size, polarity, and lipophilicity act synergistically to enhance antibiotic uptake. By defining a shared multidimensional “responsive zone,” we propose a framework to guide the selection or design of antibiotics compatible with OM-disrupting strategies, potentially enabling the repurposing of antibiotics limited by poor OM permeability. Full article

Acquired Immunodeficiency Syndrome (AIDS) is caused by Human Immunodeficiency Virus (HIV), and continues to be responsible for a substantial number of deaths worldwide each year. Development of a robust and efficient HIV-1 vaccine remains a critical priority. Structural analysis of viral proteins provides a foundational approach to designing peptide-based immunogenic vaccines. In the current experiment, we used computational prediction approaches alongside molecular docking and molecular dynamics (MD) simulations to identify potential epitopes within gp120 and Nef proteins. The selected co-epitopes were fused with the HBsAg-binding protein (SBP), a 344-amino acid protein previously identified in our laboratory through screening of a human liver cDNA expression library against HBsAg, to facilitate efficient delivery to and uptake by dendritic cells (DCs), thereby enhancing antigen (Ag) presentation. Flexible linkers are used to connect B cells, Helper T Lymphocytes (HTLs), and Cytotoxic T Lymphocytes (CTLs) in a sequential manner. The assembled vaccine construct comprises 757 amino acids, corresponding to a recombinant protein of 83.64 kDa molecular weight. Structural analysis through docking studies, MD simulations, and 3D structure validation revealed that the designed protein exhibits high structural stability and potential for interaction with Toll-like receptors (TLRs). These findings support the vaccine’s ability to enhance cellular and humoral feedback, including the stimulation of T and B cells and induction of antibody (Ab) production. The results underscore the promise of this in silico designed co-epitope vaccine as a viable candidate for HIV-1 prevention and suggest that such constructs may serve as effective immunogens in future HIV-1 vaccine strategies. Full article

Extracellular vesicles (EVs) from mesenchymal stem cells hold therapeutic promise for inflammatory and degenerative diseases; however, limited delivery and targeting capabilities hinder their clinical use. In this study, we sought to enhance the anti-inflammatory and chondroprotective effects of EVs through CAP-LAMP2b (chondrocyte affinity peptide fused to an EV membrane protein) engineering and ADAMTS4 gene editing hybrid vesicle formation. Human umbilical cord MSCs (hUCMSCs) were characterized via morphology, immunophenotyping, and trilineage differentiation. EVs from control and CAP-LAMP2b-transfected hUCMSCs were fused with liposomes carrying CRISPR-Cas9 ADAMTS4 gRNA. DiI-labeled EV uptake was assessed via fluorescence imaging. CAP-LAMP2b was expressed in hUCMSCs and their EVs. EVs exhibited the expected size (~120 nm), morphology, and exosomal markers (CD9, CD63, CD81, HSP70). CAP-modified hybrid EVs significantly enhanced chondrocyte uptake compared to control EVs and liposomes. IL-1β increased ADAMTS4 expression, whereas CAP-LAMP2b-ADAMTS4 EVs, particularly clone SG3, reversed these effects by reducing ADAMTS4 and restoring aggrecan. Western blotting confirmed suppressed ADAMTS4 and elevated aggrecan protein. CAP-LAMP2b-ADAMTS4 EVs, therefore, showed superior uptake and therapeutic efficacy in inflamed chondrocytes, attenuating inflammatory gene expression and preserving matrix integrity. These results support engineered EVs as a promising cell-free approach for cartilage repair and osteoarthritis treatment. Full article

We engineer an enhanced three-dimensional (3D) adipose model by integrating mesoporous silica (mSiO₂) nanoparticles into human adipose-derived stem cell spheroids. The mSiO₂ is highly cytocompatible, enables stable dispersion, and yields spheroids that preserve structural integrity and roundness for at least 14 days, accompanied by higher metabolic activity and reduced hypoxic stress. Under adipogenic induction, the nanoparticles embedded spheroids exhibit deeper lipid accumulation and increased expression of PPARγ, adiponectin, and FABP4. As a proof of concept, we leveraged this 3D platform to examine phage uptake and tissue-level distribution in adipose spheroids in comparison with conventional 2D cultures. These experiments reveal that both the cellular differentiation state and the tissue architecture govern phage association and uptake. Together, our findings indicate that phages engage mammalian cells beyond their bacterial hosts, a consideration that should inform future phage therapy design with implications for innate immune responses and overall therapeutic efficacy. Full article

Background/Objectives: Nivolumab has significantly improved outcomes in patients with metastatic non-small cell lung cancer (NSCLC); however, reliable prognostic biomarkers remain an unmet need. To address this gap, we developed the SUVmax-IPI, a novel prognostic index combining maximum standardized uptake value (SUVmax) from ¹⁸F-fluorodeoxyglucose positron emission tomography (FDG-PET) with systemic inflammatory markers. This study aimed to evaluate the prognostic value of SUVmax-IPI in patients with NSCLC receiving nivolumab therapy. Methods: This multicenter retrospective analysis included 187 patients with metastatic NSCLC receiving nivolumab across 5 tertiary institutions. The SUVmax-IPI incorporated pretreatment SUVmax and laboratory-based inflammatory prognostic index (IPI) parameters. Survival outcomes were evaluated using Kaplan–Meier analysis with log-rank testing and multivariate cox regression. Results: Receiver operating characteristic (ROC) analysis established an optimal SUVmax-IPI cut-off of 241.9. Patients with SUVmax-IPI ≤ 241.9 had significantly better survival outcomes: median overall survival (OS) was 35 versus 15 months (p = 0.002). For progression-free survival (PFS), although a numerical difference favored patients with SUVmax-IPI ≤ 241.9 (median: 15 vs. 8 months), this did not reach statistical significance (log-rank p = 0.175). Multivariate analysis confirmed SUVmax-IPI as an independent predictor of survival (p = 0.002). Conclusions: The SUVmax-IPI represents a promising prognostic tool for patients with metastatic NSCLC who received at least 3 months of nivolumab, integrating metabolic and inflammatory parameters to predict survival outcomes. Full article

Lipid nanoparticles are a clinically validated platform for delivering nucleic acids, but performance is constrained by multiscale physiological barriers spanning circulation, vascular interfaces, extracellular matrices, cellular uptake, and intracellular trafficking. This review links composition–structure–function relationships for ionizable lipids, helper phospholipids, cholesterol, and PEG-lipids to systemic fate, endothelial access, endosomal escape, cytoplasmic stability, and nuclear transport. We outline strategies for tissue and cell targeting, including hepatocyte ligands, immune and tumor selectivity, and selective organ targeting through compositional tuning, together with approaches that modulate escape using pH-responsive chemistries or fusion-active peptides and polymers. We further examine immunomodulatory co-formulation, route and schedule effects on biodistribution and immune programming, and manufacturing and stability levers from microfluidic mixing to lyophilization. Across these themes, we weigh trade-offs between stealth and engagement, potency and tolerability, and potency and manufacturability, noting that only a small fraction of endosomes supports productive release and that protein corona variability and repeat dosing can reshape tropism and clearance. Convergence of standardized assays for true cytosolic delivery, biomarker-guided patient selection, and robust process controls will be required to extend LNP therapeutics beyond the liver while sustaining safety, access, and scale. Full article

Oral administration remains the most patient-friendly drug delivery route, yet its efficacy is limited by physiological barriers including gastric degradation and inefficient cellular uptake. pH-responsive nanogels have shown promise for gastrointestinal drug delivery, though their effectiveness is often constrained by poor membrane interaction. Inspired by natural membrane-anchoring mechanisms, a series of comb-like anionic polymers were designed via grafting alkylamines of different chain lengths (C₁₀, C₁₄, C₁₈) at varying densities (10–30%) onto a biodegradable poly(L-lysine isophthalamide) (PLP) backbone. These pH-responsive comb-like polymers self-assembled into nanogels for loading the hydrophobic chemotherapeutic agent camptothecin. The alkyl length and grafting density significantly influenced pH-responsive behavior, membrane disruption, and drug release profiles. The optimal formulation—the nanogel prepared with PLP grafted 30% C₁₄—achieved a high drug-loading capacity, ideal particle size and stability, and offered superior protection in acidic conditions (only 7 ± 5% release at pH 1.2 over 24 h), while enabling rapid intestinal release (78 ± 2% at pH 7.4 within 24 h). The nanogels significantly enhanced cellular uptake, cytoplasmic delivery, and cytotoxicity against colorectal carcinoma cells. This study demonstrates the key role of hydrophobic modification in designing effective oral nanocarriers, providing a promising platform for the treatment of intestinal diseases. Full article

Drug delivery to the brain faces a critical obstacle in the form of the blood–brain barrier (BBB), which severely limits therapeutic options for Alzheimer’s disease (AD). Transferrin receptor 1 (TfR1) is abundantly expressed in brain capillary endothelial cells, offering a potential pathway for circumventing this barrier. Physiologically, TfR1 binds to iron-laden transferrin, leading to cellular uptake through clathrin-mediated endocytosis. Within acidic endosomes, the iron is released, and the receptor–apotransferrin complex recycles to the cell surface for further rounds of transport. Furthermore, studies in AD mouse models have demonstrated that TfR1 expression in brain microvessels remains stable, highlighting its suitability as a delivery target even in disease conditions. Based on this, various drug delivery strategies targeting TfR1 have been developed, including bispecific antibodies, antibody fragments, ligand conjugates, and nanoparticle-based carriers. While these approaches hold great promise, they face practical limitations such as competition with endogenous transferrin, receptor saturation, and inefficient intracellular trafficking. This review details the current understanding of TfR1-mediated BBB transport mechanisms, evaluates emerging delivery platforms, and argues that TfR1 represents an accessible gateway for brain-targeted therapeutics in AD. The insights presented will be of interest to researchers in molecular biology, pharmacology, and drug development. Full article

Both basic and preclinical research, as well as the development of new therapies, require tools that allow for the selective labelling of specific cell types and the targeted delivery of drugs. The developed tools must then be validated in biological systems. In view of the lack of effective therapies for many neurodevelopmental disorders, including neonatal brain injuries, we decided to use the newly described, innovative SPAchips^® (a4cell, Pozuelo de Alarcón, Spain) tool and test it in labelling neonatal rat neural cells. In our studies, rat primary cultures of neurons and glial cells (astrocytes, oligodendrocytes, and microglia) were incubated with different concentrations of SPAchips^®. At selected time points, uptake of the tested microchips by particular cell types was assessed using lineage-specific antibodies and visualized using a confocal microscope. Additionally, the potential cytotoxicity of added microparticles was verified, as was the possibility of microglia activation. The study indicates that the tested microdevices selectively label neonatal rat microglia and can be a useful tool for visualizing this cell type, as well as a non-toxic tool for developing innovative strategies based on the functionalization of microparticles aimed at modulating neuroinflammatory processes. Full article

Human amniotic epithelial cell-derived exosomes (hAECs-Exos) are nanoscale extracellular vesicles with neuroprotective, regenerative, and anti-inflammatory properties, presenting a promising cell-free therapeutic approach for ischemic stroke. This study investigated the protective effects of hAECs-Exos against ischemic injury and explored the underlying molecular mechanisms. An optimized oxygen-glucose deprivation/reoxygenation (OGD/R) model was established in murine hippocampal HT22 neurons and BV2 microglial cells to simulate ischemic conditions. hAECs-Exos were successfully isolated and characterized via transmission electron microscopy, nanoparticle tracking analysis, and Western blotting. Confocal microscopy confirmed efficient exosome uptake by both cell types. Functional analyses revealed that hAECs-Exos significantly improved cell viability, suppressed pro-inflammatory cytokine release, alleviated oxidative stress, and modulated apoptosis-related proteins. RNA sequencing identified Forkhead box protein M1 (FoxM1) as a significantly upregulated transcription factor following hAECs-Exos treatment. Further experiments demonstrated that knockdown of FoxM1 in hAECs abolished the beneficial effects of exosomes on the viability of HT22 and BV2 cells and on the suppression of inflammation, oxidative stress, and apoptosis. These findings indicate that hAECs-Exos confer neuroprotection through FoxM1-dependent mechanisms. Together, our results highlight the therapeutic potential of hAECs-Exos as a safe, effective, and clinically translatable strategy for ischemic stroke treatment, warranting future validation in vivo and rescue experiments to fully elucidate FoxM1’s causal role. Full article

Liver fibrosis, a progressive condition with limited pharmacotherapies, poses a global health challenge. Scutellarin (SCU), a flavonoid derived from Erigeron breviscapus, has demonstrated anti-fibrotic activity and modulates gut microbiota. Emerging evidence suggests that SCU may also influence the hepatic microbiome. However, its clinical utility is constrained by poor water solubility and low oral bioavailability. Here, we developed an SCU-loaded nanoemulsion (SCE) to enhance solubility and liver-targeted delivery. In vitro, SCE increased SCU uptake in hepatic stellate cells (HSCs) and significantly inhibited TGF-β1-induced fibrogenesis. In a bile duct ligation (BDL) mouse model, oral administration of SCE improved hepatic SCU accumulation and produced superior anti-fibrotic efficacy. SCE treatment attenuated fibrosis and collagen deposition in the liver and improved liver function markers. Mechanistic investigations using 16S rRNA sequencing revealed that SCU treatment was associated with beneficial microbiota changes, although its main therapeutic effects were achieved through enhanced hepatic targeting. Notably, the SCE formulation was well-tolerated, showing no significant toxicity in vitro or in vivo. In conclusion, the SCU-loaded nanoemulsion achieved enhanced hepatic delivery of SCU and exerted potent anti-fibrotic effects via multiple mechanisms, including direct suppression of fibrogenesis and ancillary modulation of the gut–liver microbiome, offering a promising therapeutic strategy for liver fibrosis. Full article