Search Results (289)

CD9 protein belongs to a family of proteins called tetraspanins, so named for their four-transmembrane-spanning architectures. These proteins are located in domains in the plasmatic membrane, called tetraspanin-enriched microdomains (TEMs). Several proteases and cellular receptors for virus entry cluster into TEMs, suggesting that TEMs are preferred virus entry portals. Severe Acute Respiratory Syndrome coronavirus 2 (SARS-CoV-2) spike (S) protein mediates virus attachment and entry into cells by binding to human angiotensin-converting enzyme 2 (ACE-2). In addition, the secretory, type-I membrane-bound SARS-CoV-2 S protein is synthesized as a precursor (proS) that undergoes posttranslational cleavages by host cell proteases, such as furin and TMPRSS2. Moreover, it has been shown that neuropilin-1 (NRP1), which is known to bind furin-cleaved substrates, potentiates SARS-CoV-2 infectivity. Our results indicate that CD9 facilitates SARS-CoV-2 infection. In addition, we show how knocking out CD9 leads to a decrease in the expression of NRP1, a protein that improves SARS-CoV-2 infection. Furthermore, we show that CD9 colocalizes with ACE-2, NRP1, furin, and TMPRSS2 at the plasma membrane; that the absence of CD9 decreases the expression of these proteins on the plasma membrane CD9-enriched microdomains, and that CD9 interacts with ACE2. In conclusion, our data suggest that CD9 facilitates SARS-CoV-2 infection and that CD9 brings together different host proteins involved in SARS-CoV-2 attachment and entry into host cells, such as ACE2, NRP1, furin, and TMPRSS2. Importantly, the fact that a blocking antibody targeting CD9 can effectively reduce SARS-CoV-2 titers highlights not only the mechanistic role of CD9 in viral entry but also offers translational potential, suggesting that tetraspanin-targeting antibodies could be developed as therapeutic agents against SARS-CoV-2 and possibly other coronaviruses, with meaningful implications for clinical intervention. Full article

Mitochondrial fission and fusion appear to be relatively infrequent in cardiac cells compared to other cell types; however, the proteins involved in these events are highly expressed in adult cardiomyocytes (ACM). Therefore, these proteins likely have additional non-canonical roles. We have previously shown that DRP1 not only participates in mitochondrial fission processes but also regulates mitochondrial bioenergetics in cardiac tissue. However, it is still unknown where the DRP1 that does not participate in mitochondrial fission is located and what its role is at those non-fission spots. Therefore, this manuscript will clarify whether oligomeric DRP1 is located at the SR–mitochondria interface, a specific region that harbors the Ca²⁺ microdomains created by Ca²⁺ release from the SR through the RyR2. The high Ca²⁺ microdomains and the subsequent Ca²⁺ uptake by mitochondria through the mitochondrial Ca²⁺ uniporter complex (MCUC) are essential to regulate mitochondrial bioenergetics during excitation–contraction (EC) coupling. Herein, we aimed to test the hypothesis that mitochondria-bound DRP1 preferentially accumulates at the mitochondria–SR contacts to deploy its function on regulating mitochondrial bioenergetics and that this strategic position is modulated by calcium in a beat-to-beat manner. In addition, the mechanism responsible for such a biased distribution and its functional implications was investigated. High-resolution imaging approaches, cell fractionation, Western blot, 2D blue native gel electrophoresis, and immunoprecipitations were applied to both electrically paced ACM and Langendorff-perfused beating hearts to elucidate the mechanisms of the strategic DRP1 localization. Our data show that in ACM, mitochondria-bound DRP1 clusters in high molecular weight protein complexes at mitochondria-associated membrane (MAM). This clustering requires DRP1 interaction with β-ACTIN and is fortified by EC coupling-mediated Ca²⁺ transients. In ACM, DRP1 is anchored at the mitochondria–SR contacts through interactions with β-ACTIN and Ca²⁺ transients, playing a fundamental role in regulating mitochondrial physiology. Full article

Nepenthes L. species (tropical pitcher plants) are a classic example of carnivorous plants. The Nepenthes traps are highly specialized pitchers with a zoned structure. On the outer surface of the pitcher, there are nectaries and various types of trichomes, including glandular trichomes. The main aim of our study was to examine these glandular trichome structures and check the distribution of the homogalacturonans (HGs) and hemicelluloses in the cell wall of trichome cells. The structure of Nepenthes bicalcarata Hook. f. and Nepenthes albomarginata T.Lobb ex Lindl. trichomes was analyzed using light and electron microscopy. The antibodies were used against the wall components [anti-pectic homogalacturonans (HGs): JIM5 (low methylesterified HGs), LM19 (low methylesterified HGs), CCRC-M38 (a fully de-esterified HGs), JIM7 (highly esterified HGs), LM20 (esterified HGs), LM5 (galactan) and anti-hemicelluloses: LM25 (xyloglucan), LM15 (galactoxyloglucan), CCRC-M138 (xylan), and LM10 antibody (xylan)]. The localization of the examined compounds was determined using immunohistochemistry techniques. The presence of endodermal and transfer cells supports the idea that peltate trichomes actively transport solutes. Also, the presence of pectic homogalacturonans and hydrophilic hemicelluloses indicates that water or aqueous solutions are transported through the trichomes’ cell walls. Our study supports the idea that these trichomes may act as hydathodes or hydropotes. Full article

Serotonin and norepinephrine transporters (SERT and NET), located on the presynaptic terminals, regulate serotonergic (5-HT) and noradrenergic (NE) neurotransmission by rapid reuptake of released amines from the synapse. Clinically used antidepressants and highly abused psychostimulants have high affinity for these transporters. The function and expression of SERT and NET are altered in mood disorders and psychostimulant use. Therefore, appropriate functional regulation of SERT and NET is important in maintaining normal homeostasis of 5-HT and NE signaling. Both SERT and NET possess kinase-specific phospho-sites/motifs and exist in phosphorylated state. Several cellular protein kinases and phosphatases regulate the dynamics of phosphorylation of SERT and NET, which in turn determine the subcellular expression and trafficking, microdomain-specific protein–protein interactionsprotein-protein interactions, transporter protein degradation and ultimately transport capacity. Dysregulations in the dynamics of SERT and NET phosphorylation and their impact on functional regulation might contribute to neuropsychiatric disorders. However, the neurobiological consequences and behavioral outcome of SERT and NET phosphorylation in vivo are not fully understood. Studies using intact animal models that directly link the phosphorylation of SERT and NET to regulatory molecular mechanisms and animal behavior are just beginning to emerge. This review summarizes our understanding of the role of phosphorylation-dependent regulation of SERT and NET in animal behaviors relevant to mood and psychostimulant use disorders. Understanding of phosphorylation-dependent molecular mechanisms of SERT and NET regulation is pivotal to identifying potential candidate mechanisms as therapeutic targets in the treatment of neuropsychiatric disorders. Full article

The search for sustainable, economically viable, and effective plant protection strategies against pathogenic bacteria, fungi, and viruses is a major challenge in modern agricultural practices. Chitosan (CS) is an abundant cationic natural biopolymer known for its biocompatibility, low toxicity, and antimicrobial properties. Its potential use in agriculture for pathogen control is a promising alternative to traditional chemical fertilisers and pesticides, which raise concerns regarding public health, environmental protection, and pesticide resistance. This study focused on the preparation of chitosan nanoparticles (CS-NPs) through cross-linking with organic molecules, such as tannic acid (TA). Various formulations were explored for the development of stable nanoscale particles having encapsulation capabilities towards low compounds of varying polarity and with potential agricultural applications relevant to plant health and growth. The solution properties of the NPs were assessed using dynamic and electrophoretic light scattering (DLS and ELS); their morphology was observed through atomic force microscopy (AFM), while analytical ultracentrifugation (AUC) measurements provided insights into their molar mass. Their properties proved to be primarily influenced by the concentration of CS, which significantly affected its intrinsic conformation. Additional structural insights were obtained via infrared and UV–Vis spectroscopic measurements, while detailed fluorescence analysis with the use of three different probes, as model cargo molecules, provided information regarding the hydrophobic and hydrophilic microdomains within the particles. Full article

Respiratory syncytial virus (RSV) is a major human respiratory pathogen, particularly affecting infants, the elderly, and immunocompromised individuals. RSV exists in both spherical and filamentous forms, with the filamentous morphology associated with enhanced infectivity and cell-to-cell spread. Here, we demonstrate that RhoA, a small GTPase involved in cytoskeletal regulation, is essential for filamentous RSV morphogenesis through its role in organizing lipid raft microdomains. Rhosin, a selective RhoA inhibitor developed through structure-guided screening, disrupts GEF–RhoA interactions to block RhoA activation. The pharmacological inhibition of RhoA with Rhosin significantly reduced filamentous virion formation, disrupted RSV fusion (F) protein colocalization with lipid rafts, and diminished cell-to-cell fusion, without affecting overall viral replication. Scanning electron microscopy revealed that Rhosin-treated infected HEp-2 cells exhibited fewer and shorter filamentous projections compared to the extensive filament formation seen in untreated cells. β-galactosidase-based fusion assays confirmed that reduced filamentation corresponded with decreased cell-to-cell fusion. The biophysical separation of RSV spherical and filamentous particles by sucrose gradient velocity sedimentation, coupled with fluorescence and transmission electron microscopy, showed that Rhosin treatment shifted virion morphology toward spherical forms. This suggests that RhoA activity is critical for filamentous virion assembly, which may enhance viral spread. Immunofluorescence microscopy using lipid raft-selective dyes (DiIC₁₆) and fusion protein-specific antibodies revealed the strong co-localization of RSV proteins with lipid rafts. Importantly, the pharmacological inhibition of RhoA with Rhosin disrupted F protein partitioning into raft domains, underscoring the requirement for intact lipid rafts in assembly. These findings highlight a novel role for host RhoA signaling in regulating viral assembly through raft microdomain organization, offering a potential target for host-directed antiviral intervention aimed at altering RSV structural phenotypes and limiting pathogenesis. Full article

In the last decade, magnetic nanoparticles (MNPs) have attracted much attention for the implementation of non-invasive approaches suitable for the diagnosis and treatment of vascular diseases. In this work, the optimization of novel vascular grafts loaded with Nickel-based nanoparticles via electrospinning is proposed. Two different polycarbonate urethanes—i.e., Corethane A80 (COT) and Chronoflex AL80 (CHF)—were used to fabricate 3D electrospun nanocomposite grafts. SEM analysis showed a homogeneous distribution of fibers, with slight differences in terms of average diameters as a function of the polymer used—(1.14 ± 0.18) µm for COT, and (1.33 ± 0.23) µm for CHF—that tend to disappear in the presence of MNPs—(1.26 ± 0.19) µm and (1.26 ± 0.213) µm for COT/NPs and CHF/NPs, respectively. TGA analyses confirmed the higher ability of CHF to entrap MNPs in the fibers—18.25% with respect to 14.63% for COT—while DSC analyses suggested an effect of MNPs on short-range rearrangements of hard/soft micro-domains of CHF. Accordingly, mechanical tests confirmed a decay of mechanical strength in the presence of MNPs with some differences depending on the matrix—from (6.16 ± 0.33) MPa to (4.55 ± 0.2) MPa (COT), and from (3.67 ± 0.18) MPa to (2.97 ± 0.22) MPa (CNF). The in vitro response revealed that the presence of MNPs did not negatively affect cell viability after 7 days in in vitro culture, suggesting a promising use of these materials as smart vascular grafts able to support the actuation function of vessel wall muscles. Full article

The L-type calcium channels (LTCCs) function as the main entry points that convert myocyte membrane depolarization into calcium transients, which drive every heartbeat. There is increasing evidence to show that maladaptive remodeling of these channels is the cause of heart failure with reduced ejection fraction (HFrEF) and heart failure with preserved ejection fraction (HFpEF). Recent experimental, translational, and clinical studies have improved our understanding of the roles LTCC expression, micro-domain trafficking, and post-translational control have in disrupting excitation–contraction coupling, provoking arrhythmias, and shaping phenotype specific hemodynamic compromise. We performed a systematic search of the PubMed and Google Scholar databases (2015–2025, English) and critically evaluated 17 eligible publications in an effort to organize the expanding body of work. This review combines existing data about LTCC density and T-tubule architecture with β-adrenergic and Ca²⁺/calmodulin-dependent protein kinase II (CaMKII) signaling and downstream sarcoplasmic reticulum crosstalk to explain how HFrEF presents with contractile insufficiency and how HFpEF shows diastolic calcium overload and stiffening. Additionally, we highlight the emerging therapeutic strategies aimed at restoring calcium homeostasis such as CaMKII inhibitors, ryanodine receptor type 2 (RyR2) stabilizers, and selective LTCC modulators without compromising systolic reserve. The review establishes LTCC dysregulation as a single mechanism that causes myocardial dysfunction while remaining specific to each phenotype, thus offering clinicians and researchers a complete reference for current concepts and future precision therapy approaches in heart failure. Full article

Poly(vinylidene fluoride-trifluoroethylene) (PVTF) nanoparticles coatings with electrically heterogeneous microdomains were engineered to mimic the natural electromechanical microenvironment of bone tissue, offering a novel strategy to enhance osteogenesis. Through a biphasic solvent phase separation method, PVTF nanoparticles (NPs) were synthesized and spin-coated onto substrates, followed by melt-recrystallization to achieve high β-phase crystallinity. The substrates were then subjected to corona poling, a process involving high-voltage corona discharge to electrically polarize and align the molecular dipoles. Structural and electrical characterization revealed tunable microdomain surface potentials and piezoelectric coefficients, correlating with enhanced hydrophilicity. Notably, microdomain potential—produced by controlled polarization—was shown to directly regulate cellular responses. In vitro studies demonstrated that a corona-poled PVTF NP coating significantly improved bone marrow mesenchymal stem cell (BMSC) proliferation and early osteogenic differentiation. This work establishes a surface electropatterning approach and highlights the critical role of electrical heterogeneity in bone regeneration, offering a novel strategy for bioactive biomaterial design. Full article

Caveolae are distinctive, flask-shaped structures within the cell membrane that play critical roles in cellular signal transduction, ion homeostasis, and mechanosensation. These structures are composed of the caveolin protein family and are enriched in cholesterol and sphingolipids, creating a unique lipid microdomain. Caveolae contribute to the functional regulation of various ion channels through both physical interactions and involvement in complex signaling networks. Ion channels localized within caveolae are involved in critical cellular processes such as the generation and propagation of action potentials, cellular responses to mechanical forces, and regulation of metabolism. Dysregulation of caveolae function has been linked to the development of various diseases, including cardiovascular disorders, neurodegenerative diseases, metabolic syndrome, and cancer. This review summarizes the ion channel function and regulation in caveolae, and their pathological implications, offering new insights into their potential as therapeutic targets for ion channel-related diseases. Full article

The excellent temperature stability and high saturation polarization of Bi_0.5Na_0.5TiO₃ (BNT) make it a promising candidate for energy storage capacitors. However, its disadvantages, such as low breakdown strength, high remnant polarization, and a complex sintering process, limit its further development. To address this, (1 − x) Bi_0.5Na_0.5TiO₃−x Sr(Mg_1/3Nb_2/3)O₃ ceramics were fabricated, where ion doping was employed to modify the domain structure, reduce the grain size, and improve the energy storage performance. With the increase in dopant concentration, the evolution from long-range-ordered ferroelectric micro-domains into short-range-ordered randomly oriented polar nanoregions (PNRs) was revealed, as demonstrated by XRD and Raman spectroscopy. This resulted in a diffuse phase transition peak and a significant reduction in remnant polarization. However, the saturation polarization also decreased. Finally, the optimal energy storage performance was achieved at a medium dopant concentration (x = 0.10), accompanied by reduced grain size and a dense microstructure. This composition exhibited a discharged energy density of 1.64 J/cm³ at a low electric field of 150 kV/cm, representing a notable improvement over pure BNT, which showed a highly lossy P-E loop and a discharged energy density of only 0.14 J/cm³ at the same electric field. Full article

Background: GM3 Synthase Deficiency (GM3SD) is a rare autosomal recessive neurodevelopmental disease characterized by recurrent seizures and neurological deficits. The disorder stems from mutations in the ST3GAL5 gene, encoding GM3 synthase (GM3S), a key enzyme in ganglioside biosynthesis. While enzyme deficiencies affecting ganglioside catabolism are well-documented, the consequences of impaired ganglioside biosynthesis remain less explored. Methods: To investigate GM3SD, we used a Human Embryonic Kidney 293-T (HEK293-T) knockout (KO) cell model generated via CRISPR/Cas9 technology. Lipid composition was assessed via high-performance thin-layer chromatography (HPTLC); glycohydrolase activity in lysosomal and plasma membrane (PM) fractions was enzymatically analyzed. Lysosomal homeostasis was evaluated through protein content analysis and immunofluorescence, and cellular bioenergetics was measured using a luminescence-based assay. Results: Lipidome profiling revealed a significant accumulation of lactosylceramide (LacCer), the substrate of GM3S, along with increased levels of monosialyl-globoside Gb5 (MSGb5), indicating a metabolic shift in glycosphingolipid biosynthesis. Lipid raft analysis revealed elevated cholesterol levels, which may impair microdomain fluidity and signal transduction. Furthermore, altered activity of lysosomal and plasma membrane (PM)-associated glycohydrolases suggests secondary deregulation of glycosphingolipid metabolism, potentially contributing to abnormal lipid patterns. In addition, we observed increased lysosomal mass, indicating potential lysosomal homeostasis dysregulation. Finally, decreased adenosine triphosphate (ATP) levels point to impaired cellular bioenergetics, emphasizing the metabolic consequences of GM3SD. Conclusions: Together, these findings provide novel insights into the molecular alterations associated with GM3SD and establish the HEK293-T KO model as a promising platform for evaluating potential therapeutic strategies. Full article

Hypertension (HTN) is a complex disease with significant global health implications, driven by neural and oxidative mechanisms. Reactive oxygen species (ROS), once considered mere metabolic byproducts, are now recognized as one of the key contributors to dysfunction of the autonomic nerve system, which involves the onset and progression of HTN. This review highlights the dynamic roles of ROS in neuronal signaling, subcellular compartmentalization, and brain–immune interactions, focusing on their impacts on synaptic remodeling, neuroinflammation, and epigenetic modifications within key autonomic regions such as the paraventricular nucleus and rostral ventrolateral medulla. We discuss novel ROS sources, including microglia-derived and endoplasmic reticulum stress-related ROS, and their contributions to HTN. Subcellular dynamics, such as ROS signaling at mitochondria-associated membranes and neuronal microdomains, are explored as activators of the sympathetic nerve system. Emerging evidence has linked ROS to epigenetic regulation, including histone modifications and non-coding RNA expression, with sex-specific differences offering insights for the development of personalized therapies. Innovative therapeutic strategies targeting ROS involve precision delivery systems, subcellular modulators, and circadian-optimized antioxidants. We propose several priorities for future research, including the real-time imaging of brain ROS, translating preclinical findings into clinical applications, and leveraging precision medicine to develop tailored interventions based on ROS activity and genetic predisposition. Through emphasizing the spatial and temporal complexity of ROS in HTN, this review identifies novel therapeutic opportunities and establishes a foundation for targeted treatments to address this health challenge. Full article

This article discusses the preparation of twin free X-cut lithium niobate wafers using the diffusion method. The liquid electrode method was used to eliminate parasitic microdomains at dislocations. According to research, the Li-rich lithium niobate polycrystalline material contains (Li_0.941Nb_0.059) Nb_0.9528O₃ and Li₃NbO₄ phases, and the diffused near-stoichiometric lithium niobate wafer exhibits a monodomain state. The piezoelectric coefficient (d₃₃) of near-stoichiometric lithium niobate after eliminating microdomains increased by 12% compared to congruent lithium niobate. The Curie temperature of near-stoichiometric lithium niobate wafers can reach 1198 °C, and the UV absorption spectrum of near-stoichiometric lithium niobate is blue shifted by 10 nm compared to congruent lithium niobate wafers, making it more suitable for fabricating electro-optic and micro nano electronics devices. Full article

Background: Sorting nexin 19 (SNX19) is important in the localization and trafficking of the dopamine D₁ receptor (D₁R) to lipid raft microdomains. However, the interaction between SNX19 and the lipid raft components caveolin-1 or flotillin-1 and, in particular, their roles in the cellular endocytosis and cell membrane trafficking of the D₁R have not been determined. Methods: Caveolin-1 and flotillin-1 motifs were analyzed by in silico analysis; colocalization was observed by confocal immunofluorescence microscopy; protein-protein interaction was determined by co-immunoprecipitation. Results: In silico analysis revealed the presence of putative caveolin-1 and flotillin-1 binding motifs within SNX19. In mouse and human renal proximal tubule cells (RPTCs), SNX19 was localized mainly in lipid rafts. In mouse RPTCs transfected with wild-type (WT) Snx19, fenoldopam (FEN), a D₁-like receptor agonist, increased the colocalization of SNX19 with caveolin-1 and flotillin-1. FEN also increased the co-immunoprecipitation of SNX19 with caveolin-1 and flotillin-1, effects that were prevented by SCH39166, a D₁-like receptor antagonist. The FEN-mediated increase in the residence of SNX19 in lipid rafts and the colocalization of the D₁R with caveolin-1 and flotilin-1 were attenuated by the deletion of a caveolin-1 (YHTVNRRYREF) (ΔCav1) or a flotillin-1 (EEGPGTETETGLPVS) (ΔFlot1) binding motif. The FEN-mediated increase in intracellular cAMP production was also impaired by the deletion of either the flotillin-1 or caveolin-1 binding motif. Nocodazole, a microtubule depolymerization inhibitor, interfered with the FEN-mediated increase in the colocalization between SNX19 and D₁R. Conclusion: SNX19 contains caveolin-1 and flotillin-1 binding motifs, which play an important role in D₁R endocytosis and signaling. Full article