Search Results (1,296)

Pepper (Capsicum annuum), a widely cultivated vegetable of significant economic importance globally, is frequently subjected to attacks from pathogens such as Ralstonia solanacearum, as well as high-temperature stress. However, the mechanisms by which pepper combats these stresses remain poorly understood. Herein, we reported that the expression of the leucine-rich repeat protein CaLRR13, which lacks a nucleotide-binding site (NBS), kinase domains, and a transmembrane region, was transcriptionally activated by both R. solanacearum inoculation and high-temperature stress. Through transient overexpression in the epidermal cells of Nicotiana benthamiana leaves, we found that CaLRR13 localized in both the cytoplasm and the nuclei. Reducing the expression of CaLRR13 via virus-induced gene silencing (VIGS) increased the sensitivity of pepper to R. solanacearum infection and high-temperature exposure, accompanied by reduced expression of immunity- and thermotolerance-related genes, including CaWRKY40, CaPR1, CaNPR1, CaDEF1, and CaHSP24. In contrast, transient overexpression of CaLRR13 in pepper leaves induced a like-hypersensitive response (HR) and enhanced the expression of the aforementioned immunity- and thermotolerance-related genes. Thus, we conclude that CaLRR13 plays a positive role in pepper immunity against R. solanacearum and thermotolerance, providing a new perspective on the crosstalk and management of plant responses to these two stresses. Full article

Globoid cell leukodystrophy (GLD) is a devastating lysosomal storage disorder caused by galactocerebrosidase (GALC) deficiency, leading to cytotoxic psychosine accumulation, broad neuroinflammation, dysfunction of autophagy and ubiquitin-proteasome system, progressive demyelination in both the central (CNS) and peripheral nervous systems (PNS), and premature death. Curative treatments are lacking, highlighting the urgent need for transformative approaches. Existing therapies have failed to achieve durable metabolic correction across neural compartments or sustained functional recovery. Here, we demonstrate that a single intracranial administration of high-titer AAV9-GALC targeting the thalamus and deep cerebellar nuclei achieves unprecedented and lifelong therapeutic efficacy in the Twitcher mouse model of GLD. This region-specific monotherapy achieved broad neuronal and glial transduction throughout the CNS and PNS, resulting in sustained supraphysiological GALC activity and complete normalization of psychosine levels. Treated mice exhibited preserved proteostasis, axonal architecture, and myelin integrity, inhibition of neuroinflammation, alongside restored motor function. Remarkably, treated mice attain lifespans approaching wild-type levels, far surpassing all previously reported interventions in this model, indicating a durable, possibly lifelong therapeutic effect. By achieving durable and comprehensive metabolic and structural correction across neural systems without repeated dosing, multi-route delivery, combinational therapy, hematopoietic stem cell transplantation, or high-dose systemic delivery, this study establishes CNS-directed AAV9 monotherapy as a clinically translatable and potentially lifelong therapeutic paradigm for GLD. Full article

Platelets and their extracellular vesicles (EVs) have emerged as promising liquid biopsy biosources for cancer detection and monitoring. The megakaryoblastic MEG-01 cell line offers a controlled system for generating platelet-like particles (PLPs) and EVs through valproic-acid-induced differentiation. Here, we performed comprehensive characterization and proteomic validation of MEG-01-derived populations, native human platelets, and their EVs using nanoparticle tracking analysis, transmission electron microscopy, imaging flow cytometry and quantitative proteomics. MEG-01 megakaryocytic differentiation is characterized by polylobulated nuclei, proplatelet formation, and elevated CD41/CD42a expression. PLPs predominantly exhibit an activated-like phenotype (CD62P+, degranulated morphology), while microvesicles (100–500 nm) and exosomes (50–250 nm) displayed size distributions and phenotypic markers consistent with native platelet-derived EVs. Proteomics identified substantial core proteomes shared across fractions and fraction-specific patterns consistent with selective cargo partitioning during EV biogenesis. Functional enrichment indicated that MEG-01-derived vesicles preserve key hemostatic, cytoskeletal, and immune pathways commonly associated with platelet EV biology. Ingenuity Pathway Analysis showed that PLPs exhibit proliferative transcriptional programs (elevated MYC/RB1/TEAD1, reduced GATA1), while plasma exosomes display minimal differential pathway activation compared to MEG-01 exosomes. Overall, these findings suggest that MEG-01-derived EVs approximate certain aspects of megakaryocyte-lineage exosomes and activated platelet-like states, although they do not fully replicate native platelet biology. Notably, plasma exosomes show strong proteomic convergence with MEG-01 exosomes, whereas platelet exosomes retain distinct activation-related features. Full article

Background/Objectives: Fumarate hydratase-deficient leiomyomas (dFH-LMs) are a rare subtype of uterine smooth muscle tumors (USMTs) with implications for hereditary leiomatosis and renal cell carcinoma (HLRCC). Although several morphologic clues have been proposed, their diagnostic reproducibility is poorly defined. We aimed to determine the diagnostic significance of histopathologic features associated with fumarase deficiency and the reproducibility of key morphologic criteria for defining dFH-LMs. Methods: A retrospective analysis was performed on 45 USMTs that were initially classified as atypical leiomyomas (ALMs). The cohort comprised patients aged 21 to 75 years who had surgery at one tertiary medical care center from 2016 to 2025. Hematoxylin–eosin (H&E) slides were independently examined by three pathologists for features associated with FH deficiency, including eosinophilic globules, staghorn-like vessels, diffuse nuclear atypia, “bizarre” nuclei, and prominent nucleoli with halos. Molecular status was determined by immunohistochemistry (IHC) for fumarate hydratase (FH) and S-(2-succino)-cysteine (2SC). Interobserver agreement was quantified using Fleiss’s κ and Cohen’s κ. Results: Loss of FH expression was detected in 26/45 tumors (57.7%). Eosinophilic globules occurred in 88.5% of dFH-LMs, but only in 15.8% of ALMs (p < 0.001). By majority consensus (≥2 of 3 reviewers), the eosinophilic globules predicted FH deficiency with a sensitivity of 88.0% and a specificity of 85.0%; interobserver reproducibility was substantial (κ = 0.63). Staghorn-like vessels occurred in 73.1% of dFH-LMs vs. 26.3% of ALMs (p = 0.02) and diffuse nuclear atypia (84.6%) was also more frequent in dFH-LMs (p = 0.01). Patients with dFH-LMs were significantly younger than those with ALMs (Median, 34 vs. 41 years). Conclusions: Eosinophilic globules, staghorn-like vessels and diffuse nuclear atypia have been shown to have high diagnostic value and could be considered morphologic indicators of dFH-LMs. The substantial interobserver reproducibility of eosinophilic globules makes this feature particularly promising for routine clinical practice. Full article

When it comes to predation, alarm signals enable individuals to assess risks and modulate their behavior accordingly. These signals, often chemical in aquatic environments, can be recognized across species boundaries and are typically released through injury-induced mechanisms in response to predation. While extensively documented in teleosts, particularly those possessing epidermal club cells, such mechanisms remain poorly understood in more basal vertebrates, such as lampreys, which possess unique epidermal structures called skein cells. The present study investigated the potential role of skein cells in the immune and alarm signaling systems of the sea lamprey (Petromyzon marinus), given their strategic location in the skin and distinctive ultrastructural characteristics, such as polarized nuclei and tonofilament-rich basal cytoplasm. Although originally misidentified as club cells, skein cells may be involved in mucus secretion and the release of compounds such as sialic acid and glycoconjugates, which provide defense against aquatic pathogens. This study employed histomorphological analysis, immunoperoxidase labeling, confocal microscopy, bioinformatics, and quantitative and statistical analysis to investigate the hypothesis that skein cells contribute to anti-predator defense via the release of alarm substances. These findings provide new insights into the evolutionary origins and functional diversity of chemical signaling in the early vertebrate. Full article

Entomopathogenic viruses offer an eco-friendly biological approach for pest control, but their relatively slow action often limits practical applications. Synergistic interactions between insect viruses and chemical pesticides can amplify their control efficacy, reduce insecticide use, and thus alleviate associated risks. Here, we evaluated the combined effects of the gomphocerinae permutotetra-like virus (GPV) and the neurotoxic insecticide imidacloprid against nymphs of Locusta migratoria. In toxicity tests, neither GPV nor imidacloprid alone caused mortality from acute toxicity after 12 h (<30%), but co-application led to marked acute synergistic toxicity, significantly increasing mortality to 87% within 12 h and 93% by 96 h. Importantly, histopathological examination revealed that the synergistic treatment caused severe midgut damage, such as disrupted or absent microvilli, extensive cellular debris in the gut lumen, cell detachment from the basal lamina, and apical displacement of nuclei. Furthermore, RNA-seq and biochemical analyses showed that the cotreatment aberrantly regulated key genes involved in peritrophic membrane integrity, substantially elevated immune responses, and disrupted energy homeostasis, which collectively led to death. These critical insights on the mechanisms underpinning the synergistic action of viral and traditional chemical agents underscore the potential of such integrated strategies to rapidly, effectively, and safely control pests. Full article

The germline-restricted chromosome (GRC) is a unique and enigmatic element found exclusively in the germ cells of passerine birds, with its function and evolutionary dynamics still largely unresolved. This study utilizes cytogenetic analysis of the Eurasian bullfinch (Pyrrhula pyrrhula) to explore the meiotic behavior of the GRC. We report the novel discovery of naturally occurring tetraploid and octoploid spermatocytes in this species. Remarkably, in these polyploid cells, the GRC exhibited normal meiotic processes, including full synapsis and recombination. Recombination was restricted to the H3K9me3-negative proximal half of the GRC bivalent, implicating a chromatin-based regulation mechanism. The standard chromosome set in the polyploid cells showed orderly chromosome synapsis. The number of recombination nodules in tetraploid and octoploid nuclei was approximately equal to the standard value for diploids multiplied by 2 and 4, respectively. These findings suggest that polyploidy does not completely hinder meiotic progression in birds and offer new insights into GRC regulation during meiosis. Full article

Background: Numerous protocols exist concerning the decellularization of the esophagus, a potential alternative to the classical surgical approach for the reconstruction of the digestive tract after esophagectomy. This systematic literature review (SLR) aimed to provide an overview of the effectiveness of the current protocols. Methods: This SLR was conducted in PubMed, EMBASE, and Scopus until September 2025. Study selection, data extraction, and quality assessment were performed by two independent reviewers according to the inclusion/exclusion criteria. Results: A total of 2494 references were screened after removing duplicates. Among these references, 26 articles were included. The large majority of studies (24/26) used Sodium Dodecyl Sulfate (SDS) or Sodium DeoxyCholate (SDC), and the most common physical method was the cannulation of the esophagus (17/26). The animal model was very heterogenous. All protocols except one showed no residual cell nuclei, with only 5/19 papers confirming a satisfactory residual amount of DNA. The assessment of the extracellular matrix (ECM)—mostly qualitative—revealed global preservation but with a systematic loss of glycosaminoglycans (GAGs). Conclusions: The decellularization of the esophagus is feasible, but the definition of the optimal protocol to achieve this goal remains difficult because of the important heterogeneity among the different studies. Full article

Cutaneous melanoma remains one of the most aggressive tumors, yet the role innate immunity plays in its progression remains poorly understood. Effector elements with high regulatory potential, capable of both promoting and inhibiting tumor growth—mast cells (MCs), are of particular interest. This includes quantitatively characterizing the interactions between tryptase-positive mast cells (MCs) with atypical Melanin—A⁺ cells and describing their spatial phenotype, in relation to the stage of cutaneous melanoma. A retrospective analysis was carried out on samples retrieved from 128 patients with cutaneous melanoma (AJCC 8th edition: IA–IIID). Histological analysis, histochemistry (toluidine blue, Giemsa), and diplex /multiplex IHC for tryptase and Melan-A were performed; as well as Fluorescence imaging, 3D reconstructions and quantitative mapping in QuPath v 0.6.0. Proximity was assessed by the nucleus-to-nucleus distance: <10 μm (contact), 10–20 μm (paracrine zone), >20 μm (out of interaction). The relative amount of MCs in the intratumoral zone was lower than in the intact dermis, with a simultaneous increase in their absolute density per mm² in the melanoma microenvironment, maximum in the peritumoral area and most pronounced at stage II. Three types of interactions were identified: (i) juxtaposition without secretion, (ii) degranulation of MCs directed to tumor cells, (iii) melanosecretion of Melanin—A⁺ cells directed towards MCs, followed by phagocytosis of melanocores. An inverse intratumoral connection between the number of MCs and the number of Melanin—A⁺ cells was noted; MCs with elongated forms, extensive contacts and polarized tryptase secretion, including granule localization near/at the nuclei of adjacent cells, were frequently observed. The obtained data indicate stage-region-dependent bidirectional cross-talk between melanin and MCs, forming tissue spatial signals, potentially useful as biomarkers and targets for personalized therapy. Full article

Clarifying the function of approximately 20,000 proteins encoded by the human genome is a key challenge in the fields of medicine and biology. However, many proteins remain uncharacterized. In this review, we introduce a challenge that uses adult T-cell leukemia/lymphoma (ATL) and proteomics to study human proteins of unknown function (PUFs). The characteristic properties of ATL cells are as follows: ATL cells (1) are infected with virus, (2) are derived from CD4⁺ T cells, (3) are generated via multi-stage carcinogenesis, (4) have flower-like nuclei, and (5) are highly infiltrative in the aggressive type. Given that ATL cells have contributed to impressive basic research, such as the discovery of HTLV-1 as a human cancer virus and interleukin-2 (IL-2) receptor α chain (IL-2Rα)/CD25, which is used for identifying regulatory T (Treg) cells, ATL cell lines could still be considered an attractive research tool. Furthermore, the “Unknome database” is useful for examining function-unknown degrees of proteins of interest using known scores based on Gene Ontology (GO) annotations and protein analysis through evolutionary relationships (PANTHER). Combining ATL proteomic data obtained by us with the “Unknome database” is expected to contribute not only to investigating the pathogenetic mechanism of ATL but also to clarifying the functions of PUFs. Full article

Platelet shortage poses a significant barrier to research and transfusion therapies because native megakaryocytes (MKs) are scarce in blood. To overcome this limitation, pluripotent stem cell–derived MKs (PSC-MKs) offer a standardized, donor-independent platform for research and therapeutic development, including disease modeling and ex vivo platelet production. Here, we report a chemically defined, feeder-free protocol to generate MKs from human pluripotent stem cells (hPSCs). The protocol combines the small molecule MPL agonist Butyzamide, macrophage colony-stimulating factor (M-CSF), and three-dimensional (3D) suspension culture, achieving high efficiency and reproducibility. Butyzamide replaced recombinant thrombopoietin (TPO), yielding comparable CD41⁺/CD42b⁺ populations and enhanced polyploidization. M-CSF accelerated nuclear lobulation and induced 4N MKs, while 3D culture increased yield, cell size, and substrate detachment. Multiple independent assays confirmed mature MK hallmarks, multi-nuclei, demarcation membranes, granules, and elevated mitochondrial respiration. Single-cell RNA sequencing outlined a continuous trajectory from early progenitors to functionally specialized MK subsets. This platform enables reliable MK supply for mechanistic studies and in vitro platelet production, advancing both basic research and therapeutic development. Full article

Mutations in genes encoding sarcomeric proteins are a common cause of cardiomyopathy and sudden cardiac death in humans. We evaluated the hypothesis that myofilament dysfunction is coupled to morphological and functional alterations of cardiomyocyte nuclei in a Tnnc1-targeted knock-in (Tnnc1-p.A8V) mouse model of hypertrophic cardiomyopathy (HCM). Tnnc1 is the gene that codes for the isoform of the Ca²⁺-regulatory protein troponin C (cTnC) that is expressed in cardiomyocytes and slow skeletal muscle fibers and resides on thin filaments of sarcomeres in those muscles. This pathogenic mutation in a sarcomere gene alters many aspects of cardiomyocyte function, including sarcomere contractility, cytoplasmic Ca²⁺ buffering, and gene expression. Analysis of myocardial histological sections and isolated cardiomyocytes from adult Tnnc1-p.A8V mouse hearts revealed significantly smaller (cross-sectional area and volume) and rounder nuclei compared to those from age-matched, wild-type control mice. Changes in nuclear morphology could not be explained by differences in cardiomyocyte size or ploidy. Isolated wild-type and mutant cardiomyocyte nuclei, which are embedded centrally within myofibrils, undergo compression during contraction of the cardiomyocyte, indicating that during each heartbeat cardiomyocyte nuclei would be mechanically deformed as well as being exposed to elevated cytoplasmic Ca²⁺. Immunoblotting analysis indicated decreased nuclear localization of cardiac troponin C and decreased histone H4 expression in Tnnc1-p.A8V mouse hearts. Next, we investigated the influence of nucleocytoplasmic transport by immunofluorescence microscopy, and we could not confirm nuclear localization of cardiac troponin C in fixed myocardial tissue from adult mice. However, cardiac troponin C could be detected in healthy human-induced pluripotent stem cell-derived cardiomyocyte nuclei. We conclude that pathological myofilament dysfunction due to a pathogenic, cardiomyopathy-associated mutation can be linked to altered protein composition of cardiomyocyte nuclei and aberrant nuclear morphology. Full article

Triarylmethyl radicals (TAMs) have recently emerged as highly effective polarizing agents in dynamic nuclear polarization (DNP) under viscous conditions, enabling substantial hyperpolarization via the solid-effect (SE) DNP mechanism even at room temperature. A comparable, though less pronounced, enhancement was observed for BDPA radicals embedded in phosphocholine-based lipid bilayers. Given the increasing interest in elucidating the structure and dynamics of biopolymers and their high-molecular-weight assemblies—such as cell membranes—this study focuses on the design, synthesis, and characterization of paramagnetic agents tailored for DNP-based structural biology. To this end, we synthesized a series of TAM derivatives functionalized with lipophilic substituents and characterized their magnetic resonance properties, including isotropic hyperfine interaction (HFI) constants on carbon nuclei and electron spin relaxation times (T₁ and T_m) at low temperatures (80 K). Echo-detected EPR spectra and electron spin echo envelope modulations (ESEEM) were recorded for novel TAM incorporated into liposomes composed of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC). These low-temperature measurements revealed that the radicals are localized either at the liposome surface or within the lipid bilayer, ensuring optimal accessibility to water molecules. Crucially, the presence of a single cholesterol moiety provides strong noncovalent anchoring within the hydrophobic core of the bilayer. Guided by these findings, we identify an amphiphilic TAM bearing a single cholesterol group and polar carboxyl functionalities as a highly promising candidate for DNP applications in membrane biology, combining efficient polarization transfer, bilayer integration, and aqueous accessibility. Full article

Background/Objectives: This study was conducted to investigate the morphological impact of carbamazepine (CBZ) coated with carbon nanodots functionalised with silver nanoparticles (CNDs@AgNPs) and metal–organic framework (MOF-5) nanoparticles on the hearts of male rats with experimental epilepsy. Methods: Seventy male Wistar rats were randomly selected for the study and divided into ten groups of seven animals each. Haematoxylin–eosin staining was performed on heart tissue, and the levels of interleu-kin-6 (IL-6) and catalase (CAT) and the oxidative stress index (OSI) were determined bio-chemically. In addition, we performed morphological measurements of the heart. Results: When the heart tissues were evaluated histopathologically in all groups, it was observed that cells with pyknotic nuclei and haemorrhagic areas increased in the heart images, especially in the PTZ group with epilepsy only. Histologically normal cardiac cells and cardiac tissue were observed in the other groups. The distance between the atria was below 10 mm only in PTZ + CBZ 50 mg/kg and PTZ + CNDs@MOF-5 25 mg/kg groups. The distance between the apex of the heart and the base of the heart was the lowest in CNDs@MOF-5 25 mg/kg and CNDs@MOF-5 50 mg/kg groups. Conclusions: PTZ-induced epilepsy causes significant histopathological changes, while cardiac tissue structure is largely preserved in the treatment groups. In our literature review, we did not find any previous studies examining the effects of carbamazepine coated with two different types of nanoparticles on the cardiac morphology in an experimental epilepsy model. Full article

The fungus Coleosporium zanthoxyli is the causal agent of leaf rust in Chinese prickly ash pepper (Zanthoxylum armatum ‘Hanyuan putaoqing’), seriously impacting its industrial development. However, little is currently known about the infection and pathogenesis of C. zanthoxyli on Z. armatum. In this study, the infection of Z. armatum by C. zanthoxyli was reported at histological and cytological levels by a fluorescence microscope and transmission electron microscopy (TEM) for the first time. Fluorescence microscopy with fluorophore Alexa 488 (WGA-FITC) stained samples revealed that the infection process comprised three distinct stages: penetration (0–1 days post inoculation, dpi), parasitic growth (3–5 dpi), and sporulation (≥7 dpi). The number of haustoria increased during the osmotic and parasitic periods and then decreased; the length of hyphae also increased rapidly and then decreased. TEM analysis during these stages demonstrated that as disease severity increased, chloroplasts and mitochondria enlarged significantly, accompanied by a marked accumulation of starch granules and osmiophilic granules. At later stages, the nuclei became irregular, the grana lamellae were blurred, and the lamellar structure was arranged disorderly, and leaf tissues were extensively colonized by fungal hyphae and haustoria, leading to cellular necrosis and distorted cell walls. Notably, the sporulation phase was characterized by dense rust spore clusters covering the leaf surface. These findings provide critical insights into the ultrastructural changes induced by C. zanthoxyli during infection, elucidating key mechanisms of rust-induced damage in Chinese prickly ash and identifying the parasitic phase as a critical window for control strategies. This study lays a foundation for further research on rust pathogenesis and the development of Chinese prickly ash targeted control strategies. Full article