Search Results (2,292)

In the last decade, notable developments have occurred regarding the application of membrane vesicles—encompassing extracellular vesicles (EVs, including exosomes, microvesicles, apoptotic bodies, and others), self-organized cellular-membrane-derived vesicles, and isolated cell-bound membrane vesicles, among others—as bioinspired drug delivery systems (DDSs). A collection of 10 papers on such advances was published in the Special Issue of Pharmaceutics entitled “Advances of membrane vesicles in drug delivery systems, 2nd Edition”. These papers investigate the Minimum Information for Studies of Extracellular Vesicles (MISEV), in vivo fluorescence imaging and tracking, in vivo specific tissue targeting, and the therapeutic application of membrane vesicles as DDSs in cancers, osteoarthritis, ocular disorders, intestinal disease, and kidney diseases. The present article briefly summarizes these related topics and provides novel insights into the research on membrane vesicles as DDSs. Full article

The Kinesin superfamily members are ATP-dependent microtubule-based motor proteins that are conserved among all eukaryotic organisms and play vital roles in diverse cellular processes, such as vesicle trafficking, mitosis and meiosis, and cytoskeletal dynamics. Here, OsKIF14.3, a kinesin-14 subfamily protein, was map-based cloned and functionally analyzed. The OsKIF14.3 gene exhibited a constitutive expression pattern. OsKIF14.3 protein localized on the microtubule and formed homodimer via the conserved Coiled Coil 1 (CC1) domain. Mutation of OsKIF14.3 altered OsSWEET11′s subcellular location from the plasma membrane into both the plasma membrane and the cytoplasm, leading to abnormal starch metabolism, excess starch accumulation in the chloroplast, broken stroma lamella and yellowish leaves in oskif14.3 mutant. These results enriched our understanding of the kinesin superfamily and leaf color regulation mechanism. Full article

Background/Objectives: This NGS-based study sought to identify novel molecular markers for prostate cancer by comparing miRNA expression in cancer and benign prostatic hyperplasia (BPH) tissues. Methods: Using high-throughput sequencing and stringent statistical criteria, the study identified eleven significantly dysregulated miRNAs (five downregulated, six upregulated) that differentiate the two conditions. Enrichment analyses linked these miRNAs to several key cancer-associated pathways, including PI3K–Akt and ErbB signaling. Results: Crucially, the protein vesicle-associated membrane protein-associated protein B (VAPB) was pinpointed as a central hub, regulated by three downregulated miRNAs (miR-143-3p, miR-221-3p, and miR-222-3p). Since VAPB has not been widely studied in prostate cancer, it represents a promising, novel candidate for both diagnosis and therapeutic targeting. Conclusions: Our NGS-based analysis revealed a distinct miRNA expression signature that differentiates prostate cancer from BPH. The downregulation of several tumor-suppressive miRNAs (with concomitant upregulation of oncogenic miRNAs) in prostate cancer may contribute to malignancy—including the de-repression of novel targets like VAPB, which we identify as a promising new biomarker and therapeutic target. Full article

It is recognized that the ionization state of amphipathic molecules can affect the curvature and arrangement of the supramolecular structure of which they are a part due to changes in their shape and interactions with neighboring molecules. The pKa influences the overall charge of the molecule and its local environment, which in turn can cause it to pack into different structures, from planar lamellar membranes to curved micelles or reversed phases. It is also recognized, though less explored, that the supramolecular structure can, in turn, affect the pKa value of the molecule. We explored this possibility with oleic acid molecules and found that the apparent pKa changed by two pH units when the surfactant was forced to remain on a flat surface, compared to the value of the aggregate in suspension, where the molecule adopts the most stable supramolecular structure for each ionization state. The pKa shifted to higher values when fatty acid was forced to form planar structures, and the pH range in which neutral and ionic species coexist (conditions under which lamellar vesicles form spontaneously) increased. Thus, we propose that it is possible to control the ionization state of molecules adsorbed onto a surface, and consequently the surface charge, by modifying surface roughness. Full article

VPS13A disease is a rare, autosomal-recessive, neurodegenerative disorder characterized by involuntary movements, orofacial dystonia, seizures, psychiatric symptoms, and the presence of spiky, deformed red blood cells (acanthocytes). The disease is caused by mutations in the VPS13A gene, which encodes the VPS13A protein (previously known as chorein). This protein is a member of the family of bridge-like lipid transport proteins, involved in bulk lipid transfer between membranes and intracellular vesicle trafficking. We describe the case of a 37-year-old woman with gait instability, semi-flexed legs, and involuntary distal muscle movements. Genetic testing was performed using next-generation sequencing (NGS), followed by molecular analysis. Fibroblasts from the patient, her mother, and a healthy control were analyzed by immunofluorescence and Western blotting. NGS identified a novel homozygous 2.8 kb deletion encompassing exons 69–70 (69–70del) of the VPS13A gene (NM_033305.3). The same variant was detected in the patient’s mother in a heterozygous state and her brother in a homozygous state. Although other deletions in the gene have been described, a comprehensive search of population variant databases and the existing literature did not reveal previous reports of this deletion. Fibroblasts from the patient, her mother and a healthy control were characterized. Functional assays showed a complete absence of the VPS13A protein in the patient’s fibroblasts. This study expands the mutational spectrum of VPS13A-linked VPS13A disease and underlines the importance of comprehensive genetic analysis in atypical cases. Full article

Acteoside (ACT), a prominent compound of the hydroxytyrosol-type phenylethanol glycoside (HPG) class, is present in plants and holds significant potential for food and pharmaceutical applications. However, the limited production of ACT in plants restricts its broader utilization. Although the biosynthetic pathways of ACT are increasingly understood, its transport mechanisms within plants remain unclear. RgMATE6, a vacuolar-type Multidrug and Toxic Compound Extrusion (MATE) transporter identified in Rehmannia glutinosa (a plant known for ACT p roduction), was selected for investigation. This study aims to elucidate the role of RgMATE6 in ACT transport and its impact on ACT biosynthesis. Our study utilized a multidisciplinary approach, including in silico analysis to predict substrate specificity, quantitative real-time PCR (qRT-PCR) to quantify gene expression, HPLC to measure HPG levels, vacuolar membrane vesicle uptake assays to validate RgMATE6 transport activity in vitro, and genetic transformation in R. glutinosa to assess its functional roles in vivo. In silico analysis identified RgMATE6 as a phenolic compound transporter, and correlation analysis revealed a strong positive association between the HPG accumulation and RgMATE6 expression in R. glutinosa. Functional validation through vacuolar membrane vesicle uptake assays in Nicotiana benthamiana confirmed RgMATE6’s role as an HPG transporter, demonstrating a significant preference for ACT. Overexpression and repression experiments in R. glutinosa further demonstrated that RgMATE6 facilitates ACT import into vacuoles and enhances its production. Additionally, tissue-specific expression analysis revealed the coordinated expression patterns between RgMATE6 and six ACT biosynthetic genes in the transgenic plants. RgMATE6 facilitates the transport and accumulation of ACT within vacuoles, and its expression might synergize with ACT biosynthesis. These findings establish a framework for improving ACT and other HPG production through targeted manipulation of plant MATE transporters. Full article

Lipid vesicles and related nanocarriers often contain two compartments, such as the inner and outer leaflets of a bilayer membrane between which amphipathic molecules can migrate. We develop a stochastic model for describing the transfer kinetics of cargo between the two compartments in an ensemble of carriers, neglecting inter-carrier exchange to focus exclusively on intra-carrier redistribution. Starting from a set of rate equations, we examine the Gaussian regime in the limit of low cargo occupation where Gaussian and Poissonian statistics overlap. We derive a Fokker–Planck equation that we solve analytically for any initial cargo distribution among the carriers. Moments of the predicted distributions and examples, including a comparison between numerical solutions of the rate equations and analytic solutions of the Fokker–Planck equation, are presented and discussed, thereby establishing a theoretical foundation to study coupled intra- and inter-carrier transport processes in mobile nanocarrier systems. Full article

Extracellular vesicles (EVs) are considered to be a promising tool in disease diagnosis. However, the clinical translation of EV-based liquid biopsy faces significant challenges due to the lack of inexpensive, rapid, and high-throughput methods of EV analysis. Bead-based platforms, combined with conventional flow cytometry, allow for the simultaneous capture and immunolabeling of EVs. In this study, we present a new approach based on the label-free isolation of EVs by tannic acid-coated superparamagnetic beads (TASPMB) combined with immunofluorescence detection of EV membrane proteins using flow cytometry. First, we tested the molecular profiling capabilities of the approach using EVs derived from human breast and colorectal cancer cell lines and from plasma of colorectal cancer patients to recognize the tetraspanin protein CD63 and the epithelial cell adhesion molecule (EpCAM). Subsequently, the developed approach was validated to identify proteins on EVs enriched with TASPMB from the conditioned media of SKBR3 and HT29 cell cultures without preliminary purification by a size-exclusion chromatography (SEC) column. The developed approach demonstrates a high capacity for isolating EVs and subsequently profiling of their membrane proteins, with a total assay time of approximately 2 h. The approach presented here can be a promising tool for rapid detection of EV membrane proteins using conventional instruments, such as flow cytometry. Full article

Integrin-mediated binding is important for the metastatic dissemination of different types of cancer cells. Snake venom disintegrins obtustatin and echistatin are potent, irreversible, and selective inhibitors of α1β1 and αvβ3 integrins, respectively. Obtustatin is one of the shortest disintegrins yet described, containing 41 amino acids. It has a similar pattern of cysteines to the other disintegrin echistatin but with a KTS motif rather than a classic RGD in its active site. A surface acoustic wave biosensor was applied to prove the molecular recognition of disintegrins by their substrates. The human erythrocyte ghost cells were immobilized at the sensors to allow for the detection of kinetic binding constants of disintegrins compared to the surface of giant unilamellar vesicles (GUVs). Obtustatin binds to the erythrocyte ghost membrane with affinity in the mid-nanomolar range (2.32 × 10^–7 M), and echistatin in the low micromolar range, which indicates specific molecular recognition for both disintegrins, but the higher response for obtustatin. The data directly confirm that disintegrins bind to the erythrocyte ghost membrane, thereby supporting the previously overlooked presence of integrins in red blood cell membranes. Full article

Reactive oxygen species (ROS), inevitable by-products of aerobic metabolism, act both as regulators of signaling pathways and as mediators of oxidative stress and aging-related damage. Protein oxidative post-translational modifications (Ox-PTMs) are recognized hallmarks of aging and metabolic decline, yet the persistence of protein oxidation under different physiological conditions, such as age and diet, remains unclear. Here, we applied proteomics to mitochondrial and membrane-enriched fractions of male Fischer 344 rat cerebrum and heart, comparing Ox-PTMs across young and aged animals subjected to ad libitum nutrition (AL) or calorie restriction (CR). We identified 139 mitochondrial and membrane-associated proteins consistently exhibiting high levels of oxidation, including tricarboxylic acid (TCA) cycle enzymes, respiratory chain subunits, ATP synthase components, cytoskeletal proteins, and synaptic vesicle regulators. Functional enrichment and network analyses revealed that oxidized proteins clustered in modules related to mitochondrial energy metabolism, membrane transport, and excitation–contraction coupling. Notably, many proteins remained persistently oxidized, predominantly as mono-oxidation, without significant changes during aging or CR. Moreover, the enzymatic activity of mitochondrial complexes was not only preserved but significantly enhanced in specific contexts, and the structural integrity of the respiratory chain was maintained. These findings indicate a dual strategy for coping with oxidative stress: CR reduces ROS production to limit oxidative burden, while protein and network robustness enable functional adaptation to persistent oxidation, collectively shaping mitochondrial function and cellular homeostasis under differing physiological conditions. Full article

Background/Objectives: Metabolic dysfunction-associated steatotic liver disease (MASLD) is highly prevalent worldwide and represents a growing healthcare challenge due to its risk of progression and association with metabolic comorbidities. Extracellular vesicles (EVs), nanosized membrane-bound particles mediating intercellular communication, have emerged as candidate biomarkers in multiple diseases. This study aimed to characterize serum EV profiles in MASLD patients, stratified into obese and lean groups using a body mass index cutoff of 23 for Asians. Methods: We enrolled 170 MASLD patients, 83 obese (median age 50, range 20–80) and 87 lean (median age 50, range 20–87), along with 57 non-MASLD controls (median age 44, range 21–86). Serum EV concentrations and particle sizes were quantified using nanoparticle tracking analysis and correlated with clinical and laboratory parameters. EV cargo proteins, including tetraspanins (CD9, CD63) and lipid droplet-associated perilipins (PLIN2, PLIN3), were assessed by Western blotting. Results: Obese MASLD patients displayed marked biochemical abnormalities, whereas lean MASLD patients showed levels comparable to non-MASLD controls. Nevertheless, serum EV concentrations were elevated in both the obese and lean MASLD groups. Importantly, in lean MASLD, EV levels correlated strongly with disruptions in lipid and glycemic homeostasis. Furthermore, a reduction in the PLIN3/CD63 ratio was observed in EVs isolated from lean MASLD patients. Conclusions: Circulating EVs are elevated in both obese and lean MASLD, but lean patients demonstrate a distinctive decrease in the EV PLIN3/CD63 ratio. These findings highlight the potential of EV profiling to uncover disease heterogeneity and to inform risk stratification in MASLD. Full article

Background/Objectives: Ochratoxin A (OTA) is a widespread foodborne mycotoxin linked to chronic kidney disease of unknown etiology. Despite evidence from animal models showing OTA accumulation in the kidney, the molecular mechanisms underlying its renal disposition in humans remain only partially understood. Here, we identify human renal transporters responsible for OTA kidney accumulation, elimination, and establish Michaelis–Menten kinetics under matched conditions to directly compare transport mechanisms. We also aim to identify inhibition potential of these transport mechanisms with common dietary polyphenols. Methods: Mammalian cells and membrane vesicles overexpressing human renal transporters were used to screen and profile the uptake and efflux of OTA. Miquelianin, (-)-Epicatechin-3-O-gallate, myricetin, luteolin, and caffeic acid were tested as potential concentration-dependent transporter inhibitors. Results: We demonstrate that OTA is a substrate for human organic anion transporter (hOAT) 1 (K_m: 2.10 ± 0.50 μM, V_max: 396.9 ± 27.0 pmol/mg/min), hOAT3 (K_m: 2.58 ± 0.83 μM, V_max: 141.4 ± 30.3 pmol/mg/min), hOAT4 (K_m: 6.38 ± 1.45 μM, V_max: 96.9 ± 18.8 pmol/mg/min), and human organic anion transporting polypeptide (hOATP) 1A2 (K_m: 37.3 ± 6.2 μM, V_max: 801.0 ± 133.9 pmol/mg/min). Among efflux transporters, OTA was transported only by human breast cancer resistance protein (hBCRP), which has minimal renal expression. While none of the uptake transporters were potently inhibited (>90%) by polyphenols at 10 μM, luteolin inhibited hBCRP-mediated transport of OTA with an IC₅₀ of 22 μM and caffeic acid stimulated hBCRP-mediated efflux with an EC₅₀ of 713.8 μM, both of which are physiologically relevant intestinal lumen concentrations. Conclusions: Our results confirm that exposure to OTA will lead to renal accumulation and increased health risks in affected populations, necessitating increased scrutiny of our food sources. Full article

Methodological fusion of materials chemistry, which enables us to create materials, with nanotechnology, which enables us to control nanostructures, could enable us to create advanced functional materials with well controlled nanostructures. Positioned as a post-nanotechnology concept, nanoarchitectonics will enable this purpose. This review paper highlights the broad scope of applications of the new concept of nanoarchitectonics, selecting and discussing recent papers that contain the term ‘nanoarchitectonics’ in their titles. Topics include controls of dopant atoms in solid electrolytes, transforming the framework of carbon materials, single-atom catalysts, nanorobots and microrobots, functional nanoparticles, nanotubular materials, 2D-organic nanosheets and MXene nanosheets, nanosheet assemblies, nitrogen-doped carbon, nanoporous and mesoporous materials, nanozymes, polymeric materials, covalent organic frameworks, vesicle structures from synthetic polymers, chirality- and topology-controlled structures, chiral helices, Langmuir monolayers, LB films, LbL assembly, nanocellulose, DNA, peptides bacterial cell components, biomimetic nanoparticles, lipid membranes of protocells, organization of living cells, and the encapsulation of living cells with exogenous substances. Not limited to these examples selected in this review article, the concept of nanoarchitectonics is applicable to diverse materials systems. Nanoarchitectonics represents a conceptual framework for creating materials at all levels and can be likened to a method for everything in materials science. Developing technology that can universally create materials with unexpected functions could represent the final frontier of materials science. Nanoarchitectonics will play a significant part in achieving this final frontier in materials science. Full article

Background/Objectives: Circulating tumor cells (CTCs) and large extracellular vesicles (LEVs) are key components of the liquid biopsy that provide minimally invasive access to tumor biology. A clinically relevant subset of CTCs coexpressing epithelial and immune markers (im.CTCs) has been described, yet the origin of this phenotype remains unclear. In this study, we investigated the cellular and molecular context underlying the emergence of immune marker expression on CTCs and LEVs. Methods: Using high-resolution immunofluorescence microscopy of patient-derived blood samples, we identified direct physical interactions between white blood cells (WBCs) and both im.CTCs and im.LEVs, exclusively in patients harboring im.CTCs. Results: In several cases, WBCs partially encapsulated CTCs and LEVs, and quantitative analysis revealed localized enrichment of immune membrane markers at the contact interface, distinguishing these events from random proximity. Proteomic profiling further identified CD4+ T cells as the predominant interacting immune cell type and confirmed the presence of CD45, CD3, and CD4 on the interacting CTCs and LEVs, matching their WBC counterparts. Conclusion: These findings support membrane transfer as a potential mechanism for the acquisition of immune markers by CTCs and LEVs and provide in vivo evidence of contact-dependent tumor-immune interactions in circulation with implications for immune modulation and clinical interpretation of the im.CTC phenotype. Full article

Aggregates of misfolded α-synuclein proteins are key markers of Parkinson’s disease. The protein α-synuclein (aSyn) is an intrinsically disordered protein (IDP) and therefore lacks a single stable 3D structure, instead sampling multiple conformations in solution. It is primarily located in presynaptic terminals and is thought to help regulate synaptic vesicle trafficking and neurotransmitter release. ASyn proteins have three domains: an N-terminal domain, a hydrophobic non-amyloid-β component (NAC) core implicated in aggregation, and a proline-rich C-terminal domain. Asyn proteins with truncated C-terminal domains are known to be prone to aggregation and suggest that understanding domain–domain interactions in aSyn monomers could help elucidate the role of the flanking domains in modulating protein structure. To this end, we used Gaussian accelerated molecular dynamics (GAMD) to simulate wild-type (WT), N-terminal truncated (ΔN), C-terminal truncated (ΔC), and isolated NAC domain (isoNAC) aSyn protein variants. Using clustering and contact analysis, we found that removal of the N-terminal domain led to increased contacts between NAC and C-terminal domains and the formation of inter-domain β-sheets. Removal of either flanking domain also resulted in increased compactness of every domain. We also found that the contacts between flanking domains in the WT protein result in an electrostatic potential (ESP) that may lead to favorable interactions with anionic lipid membranes. Removal of the C-terminal domain disrupts the ESP in a way that could result in over-stabilized protein–membrane interactions. These results suggest that cooperation between the flanking domains may modulate the protein’s structure in a way that helps maintain elongation and creates an ESP that may aid favorable interactions with the membrane. Full article