Search Results (11,702)

Natural compounds have experienced increasing clinical application, but their association with rapid-onset anaphylactoid reactions (ARs) present a significant challenge to their safe use. These ARs, clinically resembling Type I hypersensitivity, are non-IgE-mediated and involve direct mast cell activation, primarily through the human Mas-related G protein-coupled receptor X2 (MRGPRX2). We computationally screened a natural compound library for MRGPRX2 activation. A human MRGPRX2-expressing cell model was established. Cell viability assays (0–80 μM) were performed to determine appropriate drug concentrations. Compared to the controls, Baohuoside I (10 μM), along with Kaempferol-3-O-rutinoside, Epigallocatechin gallate (EGCG), Isochlorogenic Acid B, Baicalin, Andrographolide, Isorhamnetin, and Dehydroandrographolide (all at 20 μM), significantly increased intracellular calcium flux (p < 0.05) and boosted tryptase and β-hexosaminidase secretion (ELISA) (p < 0.05) in mast cells. Furthermore, the degranulation induced by these compounds was inhibited by the MRGPRX2 inhibitor Z3578 at 20 μM. Neutral red staining was employed to observe cellular morphological changes. Specific compounds capable of mediating ARs through MRGPRX2 activation on mast cells were identified. This contributes to safer and more effective drug use by elucidating the potential triggers of ARs. Full article

A Group A Streptococcus (GAS, Streptococcus pyogenes) is an exclusively human pathogen whose virulence is driven by a diverse array of surface structures, secreted toxins, and immune evasion mechanisms. Central to its pathogenicity is the M protein, a surface-anchored molecule that inhibits phagocytosis by interfering with complement deposition and binding host factors such as fibrinogen. GAS also secretes a wide range of toxins and enzymes that damage tissues and disrupt host defences. Streptolysin O and streptolysin S are potent cytolysins that lyse immune cells and contribute to tissue necrosis. Pyrogenic exotoxins (such as SpeA and SpeC) act as superantigens, triggering massive, dysregulated T cell activation and cytokine release, an underlying mechanism in streptococcal toxic shock syndrome. Additional factors like DNases and streptokinase facilitate bacterial spread by breaking down host tissue and counteracting neutrophil extracellular traps (NETs). Immune evasion is further supported by the production of enzymes that interfere with complement functions, like the cleavage of chemokines and the targeting of antibodies. Together, these virulence determinants allow GAS to cause a wide spectrum of diseases, ranging from uncomplicated pharyngitis and impetigo to invasive conditions like necrotising fasciitis and sepsis. This review provides a timely overview of the important GAS virulence factors and an update on the current vaccine landscape. Full article

The trematode Clinostomum marginatum, secretes excretory-secretory products (ESPs) which have the potential to increase the viability and antioxidant activity of probiotic strains. The aim of this study was to identify the ESP profile of C. marginatum and to evaluate its anti-inflammatory activity in RAW 264.7 macrophages, as well as its effect on the viability and antioxidant activity of a consortium of bacteria comprising Lactobacillus and/or Bifidobacterium. C. marginatum was maintained in RPMI-1640 medium for ESP collection. Anti-inflammatory activity was assessed in LPS-stimulated RAW264.7 cells treated with 800 µg/mL of ESPs, measuring cell viability, nitric oxide production, and the relative expression of pro-inflammatory cytokines (IL-6, TNF-α, INF-γ) and the COX-2 gene by qPCR. The influence of ESPs (800–1600 µg/mL) on probiotic viability and antioxidant activity was determined using MTT, DPPH, hydroxyl, and superoxide radical scavenging assays. C. marginatum showed 74% survival in vitro, and SDS-PAGE analysis revealed three major protein bands in the ESPs (47, 54, and 58 kDa). ESP treatment significantly reduced nitric oxide and the mRNA expression of pro-inflammatory markers in LPS-activated macrophages. ESPs supplemented at 1200 µg/mL optimized the growth kinetics of Lactobacillus (specific growth rate μ_L = 1.12 h⁻¹, doubling time t_d = 0.62 h) and Bifidobacterium (μ_B = 1.09 h⁻¹, t_d = 0.63 h) compared to control conditions. In conclusion, ESPs from C. marginatum exhibited significant anti-inflammatory and antioxidant effects while enhancing bacterial viability, which positions them as promising candidates for biotherapeutics agents in the management of inflammatory control and gut microbiota modulation. Full article

As synthetic biology advances, prokaryotic microorganisms have become critical platforms for heterologous biosynthesis in cell factory applications. However, conventional free enzyme systems encounter substantial challenges, including inefficient intermediate transfer, toxic intermediate accumulation, and vulnerability to temperature and pH fluctuations. Enzyme complex catalytic systems offer promising solutions to these limitations. Bacillus licheniformis, a Generally Recognized as Safe (GRAS) host with exceptional protein secretion capacity, represents an ideal chassis for enzyme complex construction. This study developed a peptide-mediated platform in B. licheniformis to enable enzyme complex self-assembly and evaluated its effects on metabolic pathway performance. Five peptide elements were screened through fusion with enhanced orange/green fluorescent proteins (eOFP/eGFP) and transglutaminase (TGase). Effective peptide pairs were identified by measuring fluorescence intensity, visualizing complex formation via laser confocal microscopy, and assessing TGase activity. Subsequently, recombinant strains expressing peptide-fused key metabolic enzymes (gadTt and KdgA) were constructed for whole-cell biotransformation using gluconate as substrate to investigate the impact of peptide-mediated enzyme complexes on pyruvate synthesis. In the fluorescent protein system, P18/D18—amphipathic peptides that drive enzyme self-assembly via intermolecular hydrophobic interactions—increased extracellular fluorescence intensity of eOFP and eGFP by 31.11% and 25.21%, respectively. The D18 peptide significantly elevated TGase activity by enhancing structural stability to over 1.3-fold that of the control. For pyruvate synthesis, the peptide-mediated enzyme complex exhibited remarkable advantages in substrate conversion rate (up to 53.08%) and thermostability, confirming the platform’s ability to enhance substrate channeling despite no optimization for absolute yield. This study established a novel peptide-mediated multienzyme self-assembly platform in B. licheniformis, providing a valuable strategy for artificial metabolic channel design in synthetic biology. Full article

Diffuse large B-cell lymphoma (DLBCL) is the most frequent B-cell type of non-Hodgkin’s lymphoma. Recent genomic studies have highlighted the importance of genetic alterations in apoptotic pathways that help malignant DLBCL cells to evade apoptosis. Apoptosis evasion by DLBCL cells is known to mediate resistance to chemotherapy. Advances in the field of regulated cell death (RCD) research have identified novel therapeutic avenues in cancer. In particular, non-apoptotic RCDs can be targeted to overcome resistance to apoptosis in cancer and ensure cell death. In this review, we have highlighted the contribution of multiple RCDs, including apoptosis, necroptosis, ferroptosis, pyroptosis, PANoptosis, NETotic cell death, autophagy-dependent cell death, cuproptosis, methuosis, or mitotic death, to normal development of B lymphocytes and DLBCL pathogenesis. We have summarized molecular mechanisms governing distinct RCDs in DLBCL, differences in cell death pathways in activated B-cell (ABC) and germinal center B-cell (GCB) DLBCL subtypes, prognostic values of RCD-related genes, and discussed the implication of RCD pathways for DLBCL treatment. Notably, the impact of RCDs goes far beyond just killing tumor cells. RCD modalities are important for orchestrating the immune response and modulating the tumor microenvironment. The current review also aims to reveal the effect of different RCDs on the tumor microenvironment in DLBCL. Most RCDs play a dual role in DLBCL, demonstrating both tumor-inducing and tumor-suppressing effects, which suggests that their targeting should be exploited with caution. Our analysis suggests that pharmacological ferroptosis induction may be the most promising RCD-targeting strategy in DLBCL. Full article

In the current situation of the Internet of Things (IoT) with its billions of interconnected devices, security in this low-resource environment is paramount. A Physical Unclonable Function (PUF) is a very useful cryptographic primitive which allows us to extract unique information from a particular device in a non-reproducible way. This allows us to use a PUF in cryptography for authentication or secret-key generation. Ring Oscillators (ROs) and Transient Effect Ring Oscillators (TEROs) are oscillating structures used in both FPGAs and ASICs to build PUFs. In this paper we present an FPGA implementation of a PUF based on what we call the “TERORO” cell (TERO + RO), which is a hybrid structure that allows us to use the different functionalities of both RO and TERO in a single building block. We assess all the possible methods of extracting bits of information from the PUF based on TERORO cells. Finally, we tested the circuit and presented experimental results in terms of its uniqueness, uniformity, and reliability. In RO-counter mode, we obtain $49.74\%$ uniqueness, $54.66\%$ uniformity, and $97.81\%$ reliability across devices, while TERO-based XOR mixing achieves $52.83\%$ uniformity, $45.79\%$ uniqueness, and $93.15\%$ reliability. The FPGA footprint is 142 LUTs, 36 registers, and 82 slices. Full article

Type 2 diabetes mellitus (T2DM) is a major metabolic disorder characterized by chronic hyperglycemia with far-reaching morbidities. Among these, diabetes-related cerebrovascular complications such as ischemic and hemorrhagic stroke, cerebral blood vessel disease, and vascular dementia are significant contributors to morbidity and mortality. Adipose tissue is a metabolically active endocrine organ that becomes dysfunctional in T2DM and communicates with distant tissues via secreted factors, including extracellular vesicles such as exosomes (EXs), phospholipid bilayer-enclosed nanosized particles. These adipocyte-derived exosomes (Ad-EXs) carry bioactive cargo, including lipids, proteins, and microRNAs that influence the function of distant organs, including the brain. Evidence indicates that Ad-EXs in T2DM are a significant risk factor for cerebrovascular complications via neurovascular impairment either directly through the adipose tissue–brain axis or indirectly by other organs. This review provides an overview of current knowledge on how Ad-EXs from different adipocyte populations contribute to cerebrovascular complications through oxidative stress, blood–brain barrier disruption, neuroinflammation, and mitochondrial dysfunction. Particular emphasis is placed on recent findings and gaps in knowledge linking diabetic Ad-EXs with brain microvascular endothelial cells that mediate neurovascular crosstalk, contributing to stroke susceptibility and cognitive decline. We also discuss the potential of Ad-EXs as biomarkers and therapeutic targets for cerebrovascular complications of T2DM. Full article

It has been well established that biofilm formation plays a critical role in the pathogenesis of various plant pathogenic bacteria. However, research on this process in Paracidovorax citrulli, the causal agent of bacterial fruit blotch (BFB) in cucurbits, remains limited. Through screening of the infection pathways of P. citrulli in sweet melon leaves, observing biofilm formation morphology at bacterial colonization sites, and detecting the activities of pathogenicity-related enzymes, this study revealed that P. citrulli readily colonizes Hami melon vascular tissues following inoculation via petiole immersion, petiole dipping, or vine injection. Dense biofilms were observed within the vascular bundles of symptomatic leaf veins. Furthermore, P. citrulli was confirmed to secrete cellulase and pectinase, with enzymatic activities increasing progressively as disease severity intensified. These findings suggest that BFB development in Hami melon is likely associated with the synergistic action of P. citrulli, biofilm-mediated occlusion of xylem vessels and hydrolytic degradation of plant cell walls, which may contribute to initial water-soaked lesions and subsequent vein-associated necrosis in leaf tissues. This study provides a theoretical foundation for further elucidation of the pathogenic mechanisms of P. citrulli. Full article

Fucoidans are natural compounds that exhibit bioactivity against age-related macular degeneration (AMD), the leading cause of central vision loss in industrialized nations. Pathological factors like oxidative stress and lipid peroxidation play vital roles in AMD pathogenesis. Lipid-induced alterations in the retinal pigment epithelium (RPE) contribute to AMD development. In this study, a commercial fucoidan from Fucus vesiculosus (FVs) was tested for its activity regarding lipid-peroxidation-related effects. The human RPE cell line ARPE-19, primary porcine RPE, and RPE/choroid explants were stimulated with erastin, acting as an inducer of lipid peroxidation, and treated with fucoidan. Effects on cell viability (tetrazolium bromide (MTT) or calcein staining), vascular endothelial growth factor (VEGF) and interleukin 8 (IL8) secretion (ELISA), reactive oxygen species (ROS), protein expression (glutathione peroxidase 4 (GPX4), CD59, and retinoid isomerohydrolase (RPE65), analyzed via Western blot), and gene expression (RT-qPCR) were investigated. FVs showed protective effects against erastin-induced reduction in viability (with a 12.7% increase in viability compared to erastin), RPE65 expression (with a 4.2-fold increase compared to erastin), and GPX4 expression (with a 2.3-fold increase compared to erastin) in primary RPE. Erastin-induced VEGF secretion was attenuated by FVs in ARPE-19 and primary RPE (with an up to 1.7-fold reduction compared to erastin). Elevated IL8 levels were reduced by FV treatment in primary RPE (with a 9.1-fold reduction compared to erastin). Induced VEGF in RPE/choroid explants was reduced by FVs (with an up to 2.9-fold reduction compared to erastin), and this reduction was correlated with slight improvements in viability. In conclusion, FVs exerted protective effects against lipid-induced stress. This study reveals further effects of fucoidans against AMD-related pathologies. Full article

Background: The pancreatic beta-cell hormone insulin regulates the metabolism of carbohydrates, as well as fats and protein. While the insulin response to a carbohydrate challenge is well defined in normoglycaemic as well as dysglycaemic (prediabetes and type 2 diabetes (T2DM)) individuals, the response of co-secreted beta-cell products (C-peptide, proinsulin and proinsulin intermediates) is less well defined. This analysis aimed to establish the expected glycaemic and pancreatic beta-cell responses to a standardised mixed meal in individuals with impaired glucose tolerance (IGT) and T2DM alongside reference ranges established in normoglycaemic individuals (NGT). Methods: A total of 743 adults (104 NGT, 85 IGT and 554 T2DM) were included, none of whom were on any anti-diabetic medication at the time of initial testing. All attended following a 10 h fast, before consuming a 500 kcal solid mixed meal (calorie contribution: 58% carbohydrates, 22% fat and 20% protein). Blood samples were collected every 30 min for the 4.5 h duration of the test for the determination of plasma glucose, insulin, C-peptide and intact and total proinsulin. Median profiles with corresponding 2.5th and 97.5th percentile lines to display the expected range were calculated and plotted for the three participant groups. Results: Median profiles with ranges over a 4.5 h meal period have been created for glucose, insulin, C-peptide and intact and total proinsulin, along with respective fasting and post-meal intervals in the three participant groups with differing glycaemic status. Conclusions: The resulting profiles and ranges allow for comparison in responses to a carbohydrate challenge in individuals across the glycaemic spectrum. Full article

Hepatic osteodystrophy (HOD) is a frequent complication of chronic liver disease, marked by impaired osteogenesis and elevated fracture risk, particularly under sustained alcohol exposure. Bone morphogenetic proteins (BMPs), which play a crucial role in maintaining bone homeostasis, are dysregulated in alcoholic liver disease. Specifically, decreased BMP2 and increased BMP13 have been linked to impaired osteogenesis and cartilage-like shifts in bone progenitors. A human in vitro system that recapitulates this hepatic BMP imbalance is needed to dissect mechanisms and identify targets. To address this, we established a long-term human three-dimensional liver–bone co-culture model that integrates hepatocytes (HepaRG), hepatic stellate cells (LX-2), and human umbilical vein endothelial cells (HUVECs) with bone scaffolds seeded with osteoblast precursors (SCP-1) and osteoclast precursors (THP-1). This study aimed to characterize the effects of chronic 50 mM alcohol exposure on hepatic fibrogenic activation and BMP ligand secretion, and to investigate the associated BMP-responsive signaling involved in bone cell lineage differentiation and functional activity. The results demonstrated alcohol-induced hepatic CYP2E1 activation and fibrogenic remodeling with EMT signatures, as well as a decrease in BMP2 and an increase in BMP13, without affecting BMP9. Liver-derived factors activated both canonical and non-canonical BMP signaling in bone progenitors, reduced osteoblast activity and mineralization, preserved osteoclast TRAP activity, and shifted the lineage toward chondrogenesis (SOX9↑, RUNX2↓). Notably, this BMP profile and skeletal phenotype reflect clinical observations in chronic liver disease, indicating that the model recapitulates key in vivo pathological features. This human liver micro-organoid co-culture reproduces alcohol-induced hepatic BMP dysregulation and downstream bone defects, offering an organoid-centric, microengineered platform for mechanistic studies and BMP-targeted therapeutic screening in HOD. Full article

Background/Objectives: Uveal melanoma (UM) is the most common intraocular malignancy in adults. Although brachytherapy of the primary tumor provides an approximate 80% five-year survival, with time, nearly half of patients experience predominant liver metastases. It was proposed that malignant cells migrate early and stay dormant as they adapt to the liver microenvironment. We propose that cancer vaccine-mediated activation of UM-targeted immunity in primary UM patients could prevent progression of metastatic disease from dormant cells or malignant seeds. Thus, this study explored DNA vaccination as a measure to educate the immune system to recognize the most common UM-associated Q209L tumor driver mutation in GNAQ and GNA11 G-alpha proteins. Methods: Several DNA constructs encoding mutated GNAQ were developed and tested for activation of UM-reactive T cells in HLA-A2/Hd transgenic mice and human T cells ex vivo. Results: Constructs containing immune-enhancing PADRE and VP22-derived epitopes boosted T cell responses against mutant GNAQ, which correlated with reduced experimental lung metastases. Ex vivo dendritic cell-mediated T cell activation with vaccine constructs containing optimized structure produced cytolytic T cells that secreted IFN gamma and killed mutated GNAQ-expressing UM cells in vitro. Conclusions: These findings propose the utility of the fusion DNA vaccines in eliciting T cell immunity against UM cells bearing the Q209L mutation in GNAQ/GNA11 protein to prevent the establishment and progression of metastatic disease. Full article

A protocol was developed for the isolation and characterization of extracellular vesicles (EVs) from Coffea arabica cell suspension cultures (CSCs). The isolation method involved differential ultracentrifugation of the CSC filtrate, yielding two fractions: the pellet after 100,000×g for 36 min (100k×g) and the pellet obtained from the previous supernatant after 125,000 g for 6 h (125k×g). Both fractions were characterized by size, morphology, and proteomic profiles (ProteomeXchange identifier PXD071909). While no significant differences in average EV size were observed between the two fractions, proteomic analysis revealed distinct quantitative and compositional variations. The 100k×g fraction was enriched in proteins associated with cell periphery, plasma membrane, and extracellular region, whereas the 125k×g fraction predominantly contained proteins from the extracellular region. Proteomic marker analysis confirmed that both fractions contained protein EV markers, such as transmembrane and transport proteins, soluble EV-associated proteins, and proteins targeted to the extracellular environment or cell wall. Conversely, negligible contamination from non-EV-related proteins was detected. Furthermore, transmission electron microscopy (TEM) showed that the average size of the fractions was consistent with that reported for plant EVs. These findings demonstrate that the protocol utilized to isolate EVs from coffee CSC applies to the release of such vesicles without mechanical harsh grinding that leads to tissue/cell rupture and consequent contamination by other cell components. EVs obtained from coffee CSC represent a valuable and scalable platform, paving the way for the development of tools for biotechnological applications. Full article

Background/Objectives: Given the lack of clarity regarding how maternal overnutrition during pregnancy regulates offspring metabolic health, our study intends to explore the specific influences of maternal Western diet (WD) exposure on neonatal islet cell development and heterogeneity. Methods: Using a WD-induced gestational diabetes mellitus (GDM) rat model, we assessed glucose homeostasis via blood glucose and serum insulin levels. Target protein expression and islet function were evaluated using immunofluorescence and insulin secretion assays, respectively. To delineate alterations in cellular heterogeneity, we subsequently performed single-cell RNA sequencing (scRNA-seq) on isolated islet cells. Results: Maternal WD exposure induced significant glucose intolerance and insulin resistance, confirming GDM establishment. Their neonatal offspring consequently displayed disrupted glucose homeostasis, characterized by concurrent hypoglycemia, hyperinsulinemia, and enhanced insulin secretion. ScRNA-seq analysis further identified the enhanced endocrine cells in GDM-offspring islets, with imbalanced α/β-cell subsets—specifically, reduced immature α1/β1 subsets and expanded mature α2/β2/β3/β4 subsets, alongside upregulated expression of insulin- and glucagon-related genes (Ins1, Ins2, Gcg). Notably, β cells in GDM offspring displayed metabolic hyperactivity (enriched ribosomal and glycolytic pathways) with multiple organelle dysfunction, including mitochondrial swelling, cristae reduction, decreased membrane potential, and severe endoplasmic reticulum stress. Conclusions: The metabolic dysregulation of WD-induced GDM in maternal rats is transmitted to offspring, leading to disrupted neonatal α/β-cell subset balance and accelerated islet maturation. However, such excessive development comes at the cost of organelle damage in β cells. Our findings provide a molecular basis for mitigating the intergenerational transmission of diabetes through early nutritional interventions. Full article

Pancreatic β-cell dysfunction is the key driver of type 2 diabetes, and anthocyanins have been proposed as dietary compounds that may help preserve β-cell health. This systematic review aimed to synthesise evidence on the direct effects of anthocyanins on β-cell viability, apoptosis, oxidative stress, and insulin secretion across in vitro models. Four databases were searched in March–April 2025, and eighteen studies met the inclusion criteria. Purified anthocyanins—including cyanidin-3-glucoside (C3G), cyanidin-3-rutinoside (C3R), malvidin-3-glucoside (M3G), and delphinidin-3-glucoside (D3G)—as well as anthocyanin-rich berry extracts, were tested in INS-1, MIN6, RIN-m5F cells and primary mouse or human islets under glucotoxic, lipotoxic, oxidative, cytokine, and amyloidogenic stress. Anthocyanins consistently improved β-cell viability, reduced apoptosis, and lowered reactive oxygen species (ROS), nitric oxide (NO), and thiobarbituric acid reactive substances (TBARSs) levels while enhancing antioxidant enzyme activities. Multiple studies showed upregulation of insulin secretion-related genes and proteins, and both acute and chronic treatments increased glucose-stimulated insulin secretion under normal and stressed conditions. Mechanistic pathways involved modulation of mitogen-activated protein kinase (MAPK) signalling, endoplasmic reticulum (ER) stress responses, inflammatory mediators, and mitophagy (PINK1/PARKIN). While effective in vitro concentrations were higher than typical circulating levels, the collective evidence highlights anthocyanins as promising β-cell protective agents and underscores the need for studies examining their metabolites and physiologically relevant exposure. Full article