Search Results (342)

The ATP-binding cassette (ABC) transporter family is one of the oldest conserved protein families and is widely present in animal and plant cells. However, few studies have investigated the role of ABC in the forest musk deer (FMD; Moschus berezovskii). In this study, we employed bioinformatics methods to identify and analyze the ABC transporter genes in M. berezovskii to elucidate the potential function of ABC genes in musk secretion. A total of 51 members of the MbABC gene family were identified. The analysis encompassed various aspects including physical and chemical properties, phylogenetic tree, structure prediction, conserved motifs, gene structures, chromosome localization, collinearity analysis, and KEGG and GO enrichment. Collinearity analysis revealed that the ABC transporter gene family is conserved in FMD, Cervidae, and five Bovinae species. MbABCB6, MbABCD4, MbABCF3, and MbABCG5 are key genes in protein–protein interaction networks, which are primarily involved in the transport of vitamins, lipids, and proteins. Tissue expression analysis showed that MbABCs were expressed at different stages. The RT-qPCR analysis revealed that 12 MbABC genes were up-regulated in musk gland cells during the non-secretion phase and stimulation phase, particularly MbABCC4d and MbABCC3. This study provides comprehensive information on the ABC gene family in FMD which can be further used for their functional validation. Full article

Extracellular vesicles (EVs) secreted by Fusarium oxysporum play an important role in the process of its infestation of the host, but the in vitro research system for EVs of F. oxysporum (Fo-EVs) has not yet been improved, and the mechanism of its action remains unclear. In this study, particle size distribution, particle concentration, number of particles per unit of protein, number of particles per unit of mycelial biomass, and concentration of contaminated proteins were used as indicators to evaluate the yield and purity of Fo-EVs. The optimal method for Fo-EV preparation and extraction was screened by comparing liquid culture, solid culture, and solid culture with enzymatic cell wall hydrolysis. The optimal system for Fo-EVs separation and purification was screened by a pairwise combination of three primary methods (Ultracentrifugation (UC), Ultrafiltration (UF), and Polyethylene glycol precipitation method (PEG)) and two secondary methods (Size-exclusion chromatography (SEC) and Aqueous two-phase system (ATPS)), respectively. The protein composition was identified via mass spectrometry technology, followed by GO annotation and GO enrichment analysis using whole-genome proteins as the background. Based on these steps, a Fo-EV protein library was constructed to reveal Fo-EV’s most active biological functions. The results showed that solid culture combined with the UC-SEC method could effectively enrich Fo-EVs with a typical cup-shaped membrane structure. The obtained Fo-EVs had an average particle size of 253.50 nm, a main peak value of 200.60 nm, a particle concentration of 2.04 × 10¹⁰ particles/mL, and a particle number per unit protein of 1.09 × 10⁸ particles/μg, which were significantly superior to those of other combined methods. Through proteomic analysis, 1931 proteins enriched in Fo-EVs were identified, among which 350 contained signal peptides and 375 had transmembrane domains. GO enrichment analysis revealed that these proteins were mainly involved in cell wall synthesis, vesicle transport, and pathogenicity-related metabolic pathways. Additionally, 9 potential fungal EV markers, including Hsp70, Rho GTPase family, and SNARE proteins, were screened. This study constructed an isolation system and a marker database for Fo-EVs, providing a methodological and theoretical basis for in-depth analysis of the biological functions of Fo-EVs. Full article

NLRP3 inflammasomes are transient large protein aggregates involved in the regulation of the innate immune response but are also associated with endothelial dysfunction during vascular inflammation. While NLRP3 inflammasome assembly and activation is well characterized in immune cells, its role in endothelial cell function remains incompletely understood. This study analyses the function of promyelocytic leukemia (PML) protein, a nuclear scaffold protein that forms so-called PML nuclear bodies (PML-NBs), in the regulation of NLRP3 inflammasome activation in endothelial cell cultures. Following LPS priming and subsequent ATP-induced activation, PML played a dual role: 1. It enhanced NF-kB-dependent transcription of inflammasome components (NLRP3, pro-caspase-1 and pro-IL-1β). 2. At the same time, a post-translational reduction in NLRP3 protein levels and reduced ASC oligomerization were observed, leading to impaired inflammasome activation, as evidenced by lower caspase-1 activity and reduced IL-1β secretion. Proper formation of PML-NBs was critical for this regulatory effect on NLRP3 inflammasome formation, as PML-NBs retained ASC in the nucleus and post-translationally modified NLRP3, presumably affecting its stability. Taken together, these findings suggest that PML represents a regulatory checkpoint in endothelial inflammasome activation, preventing excessive inflammatory responses that could contribute to vascular dysfunction associated with chronic inflammation. Full article

Background: Cardiovascular diseases, driven by platelet hyperactivation and thrombosis, remain the leading global cause of death. Excessive platelet activation contributes to atherosclerosis and thrombo-inflammatory disorders, underscoring the urgent need for safer and more effective antiplatelet agents. Objectives:Xanthium strumarium L. (X. strumarium) has been reported to exhibit a wide range of pharmacological effects, including anti-inflammatory and antioxidant activities. However, its antiplatelet and antithrombotic effects remain unexplored. Therefore, the present study aimed to comprehensively evaluate the antiplatelet and antithrombotic effects of X. strumarium through integrated in vitro and in vivo experiments. Methods: The principal bioactive compounds present in the X. strumarium extract were identified through GC–MS analysis. In vitro antiplatelet effects were evaluated via light transmission aggregometry, scanning electron microscopy (SEM), ATP and calcium mobilization assays, αIIbβ3 binding assay, clot retraction assay, and Western blotting. In vivo ferric chloride-induced (FeCl₃) murine thrombus model was established to evaluate thrombogenesis. Results: Our results demonstrated that X. strumarium at 25, 50, or 100 μg/mL significantly inhibited collagen, ADP, U46619, and thrombin-induced platelet aggregation. SEM revealed that X. strumarium pretreatment markedly preserved the resting platelet morphology and inhibited collagen-induced activation and shape changes. Further, the granule secretion, integrin-αIIbβ3 signaling, and the MAPK and PI3K/Akt pathways were also concentration-dependently inhibited. The in vivo blood flow rate and mice survival were improved, and H&E staining further revealed a concentration-dependent prevention of arterial occlusion following X. strumarium treatment. Conclusions: Collectively, X. strumarium demonstrated potent antiplatelet and antithrombotic effects, improving blood flow and survival while preventing arterial occlusion. Full article

Mitochondria are essential for β-cell function, coupling glucose metabolism to ATP production and insulin secretion. In diabetes, β-cell mitochondrial dysfunction arises from oxidative stress, impaired quality control and disrupted dynamics, leading to reduced oxidative phosphorylation, defective insulin release and progressive cell loss. Key transcriptional regulators link genetic susceptibility to mitochondrial dysfunction in both type 1 diabetes mellitus (T1DM) and type 2 diabetes mellitus (T2DM). These disruptions impair mitophagy, mitochondrial translation and redox homeostasis. Therapeutic strategies that restore mitochondrial function, including mitophagy enhancers, mitochondrial antioxidants, and transcriptional regulators, have shown potential in preserving β-cell integrity. As mitochondrial failure precedes β-cell loss, targeting mitochondrial pathways may represent a critical approach to modifying diabetes progression. Full article

Background: The golden era of antibiotics has long passed, and the clinical failures caused by emerging drug-resistant bacteria have intensified the demand for novel antimicrobial agents. Antimicrobial peptides have attracted significant attention as promising candidates for next-generation antibiotics. Methods: In this study, we identified a novel antimicrobial peptide, Caerin 1.1-LC, from the skin secretion of the Australian green tree frog, Litoria caerulea. Subsequent structure–activity relationship studies led us to design a series of analogues and revealed the critical role of the peptide’s intrinsic hinge structure in shaping its biological activity. Results: Incorporation of D-isomers at the valine residues within the hinge preserved overall helical content but altered the hinge conformation, resulting in an 8-fold increase in antibacterial activity against Gram-negative bacteria. Simultaneously, haemolytic activity was markedly reduced, leading to a 56-fold improvement in therapeutic index (from 0.47 to 26.6). Structural modulation of the hinge also switched the mechanism of action from classical membrane disruption with associated permeability changes to a non-membrane-permeabilising, ‘cell-penetrating-like’ behaviour, inducing membrane potential depolarisation and ATP disruption to trigger bacterial death. In vivo studies using infected larval models, along with in vitro LPS neutralisation assays, further demonstrated the therapeutic potential of the D-analogue as a novel antibacterial agent. Conclusions: This work highlights the pivotal role of hinge structures in Caerin-family/hinge-containing AMPs, offering a strategic avenue for optimising antibacterial efficacy. Full article

Semaphorin3A (Sema3A) is a regulatory protein found to be expressed on regulatory T and B cells and also secreted into peripheral blood. It has been identified as a potent immune regulator; however, not all its regulatory mechanisms have been evaluated. In this respect, we aim to investigate how Sema3A affects key metabolic pathways in T cells during homeostasis and rheumatoid arthritis (RA), and on the AKT/mTORC1 signaling axis. In this study, peripheral blood samples were collected from 119 healthy donors and 32 rheumatoid arthritis patients. T cells were subjected to Seahorse analysis to evaluate OXPHOS and glycolysis, live cell TMRE staining to evaluate mitochondrial activity, mass spectrometry for metabolite profiling, ATP determination to study ATP production, and Western blot analysis to investigate the signaling pathway activity. This study presents evidence showing that Sema3A inhibits the AKT/mTORC1 pathway, leading to a decreased glucose uptake and glycolysis disruption. Furthermore, we show that Sema3A reduces mitochondrial capacity and OXPHOS in activated T cells of healthy and RA donors, leading to a decreased ATP production. In contrast, Sema3A upregulates fatty acid oxidation (FA), probably as a backup pathway to ensure cell survival. Results with p values of <0.05 were considered significant. Our data may point to Sema3A’s ability to convert activated T cells’ metabolic profile back to its non-activated state. This may suggest that Sema3A might be a beneficial treatment for immune-mediated diseases by metabolically reprogramming activated T cells. Full article

The P2X7 receptor has been associated with the neurodegeneration of retinal ganglion cells (RGCs), which is central to the loss of vision in glaucoma. Furthermore, the activation of P2X7 has been shown to cause the death of RGCs, including in the human retina. Human organotypic retinal cultures (HORCs) were used to investigate the potential indirect mechanisms of RGC death. Of the 27 cytokine/growth factors assayed, the stimulation of P2X7 using BzATP (100 µM; 36 h) significantly increased the secretion of IL-1β and IL-10. IL-1β was selected for further investigation. BzATP (100 µM) caused an increase in the expression and release of IL-1β in a time-dependent manner; this increase was inhibited through a co-incubation with BBG (1 µM). Exogenous IL-1β alone (10 ng/mL) did not cause a loss of RGCs. However, IL-1β inhibited the loss of RGCs caused by BzATP, and this neuroprotection was prevented by the Interleukin-1 receptor-1 antagonist (IL-1ra) (100 ng/mL). The IL1 receptor IL-1R1 was localised to the inner retina close to the RGCs, although not predominantly co-localised with RGC bodies. The results suggest that the P2X7-mediated death of RGCs is not IL-1β mediated. Furthermore, IL-1β may be upregulated as part of a response to mitigate P2X7-mediated damage to the retina. Our research is the first to indicate the P2X7-mediated regulation of IL-1β in the human retina and supports the role of the ATP/P2X7/IL-1β axis in RGC survival and possible glaucomatous RGC degeneration. Full article

In this review we propose the hypothesis that an energy-dissipating process precedes the continuous stimulation of insulin secretion by glucose. This process is mediated by connexin 36 hemichannels (Cx36H), or Cx36 connexons. Cx36H oligomers are expressed at the plasma membrane, and their gating activity (opening) is activated by plasma membrane depolarization after the closure of K⁺ATP channels by glucose (>5 mM) metabolism. This initial depolarization (1st step) might be responsible for the first phase of insulin secretion, with the subsequent opening of Cx36H increasing β-cell plasma membrane permeability, allowing for the efflux of metabolites (less than 1KD) (GABA, adenine nucleotides) and K⁺ (2nd step). This provokes a breakdown of oxidative glucose metabolism and the repolarization of the plasma membrane. As the extracellular glucose concentration increases further (>>5 mM), it exerts a progressive inhibition effect on Cx36H opening, allowing for the continuous stimulation of insulin secretion (3d step, second phase,). The glucose feature of regulating Cx36H closing with sigmoidal kinetics (8 mM IC₅₀ and around 20 mM at maximum) has been confirmed in mouse Cx36 connexin expression in Xenopus oocytes and in mouse islets stimulated by a range of glucose concentrations in the presence of 70 mM KCl. This gating activity was also inhibited by some non-metabolized glucose analogs. Glucose inhibition of Cx3H opening might not only contribute to making the insulin secretory response more specific for glucose but might also play a role in the pulsatility of sustained insulin secretion. Cx36H opening also offers the opportunity to potentiate the secretory effect in vivo by, permeant or not, metabolic stimuli. Confirmation of this novel physiological role for Cx36H in β-cells would place them as new susceptibility locus for type 1 and type 2 diabetes, whose physiological implication in the mechanism of insulin secretion regulation should be evaluated by in vivo studies in diabetic patients. Full article

p-Benzoquinone (PBQ), a highly toxic compound, is the main active component in larval oral secretions of red palm weevil (RPW), Rhynchophorus ferrugineus, playing critical roles in external immunity and pathogen defense. In this study, we demonstrated that pathogens effectively induce RPW larval external immune responses. On this basis, the toxicity of PBQ to third-instar larvae was determined, with poisoning symptoms observed. The differences in gene expression between larvae before and after treatment with PBQ were analyzed by transcriptome sequencing to potentially involve the mechanisms of PBQ toxicity on larvae and the mechanisms of detoxification in the infected larvae. The results indicated that PBQ exposure was associated with altered expression of chitinase (CHI) and phenoloxidase (PO) genes in RPW larvae, which not only affects the digestion and degradation of the old cuticle but also activates phenoloxidase, further oxidizing tyrosine for its conversion into DOPA and dopamine, resulting in the generation of melanin and different degrees of cuticular melanization. The transcriptional changes further suggest that RPW larvae may employ metabolic processes to counteract the external immune-active compound PBQ toxicity by regulating the expression levels of detoxifying enzyme-encoding genes, such as cytochrome P450 (CYP450), glutathione S-transferase (GST), and ATP-binding cassette transporter (ABC). Our research provides potential novel strategies for pest control by targeting insect metabolic detoxification systems. Full article

P2Y₂ receptors are a subclass of G protein-coupled receptors activated by the extracellular nucleotides ATP and UTP. These receptors are widely expressed in multiple tissues—including the brain, lungs, heart, and kidneys—and play pivotal roles in inflammation, wound healing, and cell migration. Through coupling with various G proteins, P2Y₂ receptors initiate diverse intracellular signaling pathways that mediate calcium mobilization, cytokine release, and cytoskeletal reorganization. Recent studies highlight their dual roles in health and disease. In physiological contexts, P2Y₂ receptors contribute to immune modulation and tissue repair. In pathological conditions, they are implicated in Alzheimer’s disease by promoting non-amyloidogenic processing of amyloid precursor protein and in dry eye disease by enhancing mucin secretion while modulating ocular inflammation. They also influence chloride secretion and mucosal hydration in cystic fibrosis and contribute to inflammatory regulation and epithelial repair in inflammatory bowel disease. Additionally, P2Y₂ receptors modulate breast cancer progression by regulating cell adhesion, migration, and matrix remodeling. Their involvement in blood pressure regulation via epithelial sodium channel modulation and their facilitative role in HIV-1 entry further underscore their clinical significance. These multifaceted functions position P2Y₂ receptors as promising therapeutic targets for diverse diseases, warranting further investigation for translational applications. Full article

Background/Objectives: Tea, the world’s second most consumed beverage after water, contains diverse phytochemicals that have garnered growing interest for their potential ability to modulate inflammasome activation. This study examined the antioxidant and anti-inflammatory properties of oolong tea (OLT) extracts, with a specific focus on their regulatory effects on NLRP3 inflammasome assembly—a critical mediator in chronic inflammatory diseases. Methods: OLT extracts were prepared from the Jin-Xuan cultivar with quantification for bioactive components (total phenolics, flavonoids, condensed tannins, and proanthocyanidins). J774A.1 murine macrophages were primed with LPS and stimulated with ATP to induce inflammasome activation. Therapeutic potentials of OLT extracts were assessed by measuring cytokine secretion, expression of NLRP3 inflammasome-related proteins (NLRP3, ASC, Caspase-1, and IL-1β), inflammasome complex formation, and ROS generation via biochemical assays, immunoblotting, and fluorescence microscopy. Results: OLT extracts, particularly at 100 µg/mL, markedly suppressed both the priming and activation phases of NLRP3 inflammasome formation. OLT treatment reduced IL-1β secretion by more than 50%, attenuated ASC oligomerization and speck formation, inhibited caspase-1 cleavage, and lowered intracellular ROS levels by approximately 50%. Conclusions: These findings suggest that OLT extracts exert potent anti-NLRP3 inflammasome activity and offer immunomodulation potential in preventing inflammation-related diseases such as infections, cancer, and neurodegenerative disorders. Further in vivo investigations, followed by clinical applications and epidemiological studies, are warranted to validate these preventive effects in human populations. Full article

Mammals exhibit unique and highly variable mechanisms for the establishment and maintenance of pregnancy. Ruminants (e.g., sheep, cows, and goats) have novel mechanisms whereby the conceptus (embryo and its extra-embryonic membranes) signals for the establishment of pregnancy and exhibits unique metabolic pathways favoring conceptus development. Embryos of ruminants reach the spherical blastocyst stage at 5 to 10 mm in diameter and then elongate rapidly to elongated filamentous conceptuses of greater than 250 mm as they make contact with the uterine luminal epithelium (LE) for implantation. During conceptus elongation the trophectoderm cells secrete interferon tau (IFNT), a novel pregnancy recognition signal for ruminants to ensure maintenance of a functional corpus luteum (CL) to secrete progesterone (P4) required for pregnancy. P4 induces uterine epithelia cells to express the endogenous Jaagsiekte Retrovirus (enJSRV) that may transactivate toll-like receptors 7 and 8 in the conceptus trophectoderm to induce secretion of IFNT, a classical viral–antiviral mechanism. IFNT silences expression of receptors for estradiol (E2) and oxytocin (OXTR), which abrogates the mechanism whereby oxytocin from CL and posterior pituitary would otherwise induce large pulses of prostaglandin F_2α (PGF) by uterine epithelia to cause regression of the CL and its secretion of P4. IFNT has another novel role in silencing expression of not only ESR1 and OXTR, but all classical interferon-stimulated genes in the uterine LE and superficial glandular epithelium (sGE), but with P4 increasing expression of genes for transport of nutrients such as glucose and arginine into the uterine lumen to support conceptus development. Ruminant conceptuses convert glucose to fructose, a novel hexose sugar that cannot be transported back to the maternal circulation. Fructose is converted to fructose-1-PO4 for metabolism, not via the pathway for glycolysis but via the novel fructolysis pathway uninhibited by low pH, citrate, or ATP as is the case for glycolysis. Thus, fructose and its metabolites support the pentose cycle, hexosamine biosynthesis pathway, one-carbon metabolism, and the citric acid cycle for all cells of the conceptus. Arginine is another key nutrient transported into the uterine lumen by the uterine LE/sGE in response to P4 and IFNT. Arginine is metabolized to generate nitric oxide, polyamines, and creatine, essential for conceptus growth and development, while enhancing production of IFNT as a novel pregnancy recognition signal, and upregulating expression of genes in the uterine LE/sGE for transport of nutrients. Fructose is the major hexose sugar supporting major metabolic pathways required for conceptus growth and development in ruminants. Full article

Giant pandas (Ailuropoda melanoleuca), a flagship endangered species under priority preservation in China, remain poorly understood in terms of their testicular physiology and the mechanisms underlying spermatogenesis. Testicular peritubular cells (TPTCs), a crucial somatic cell type surrounding seminiferous tubules, secrete growth factors such as GDNF and CSF1 and release inflammatory factors such as IL-6 and IL-1β, contributing to the testicular niche and immune homeostasis. The contraction of TPTCs also facilitates the transport of sperm towards the epididymis. Nonetheless, TPTCs tend to undergo replicative senescence in vitro, which is a hinderance to their in-depth study. Here, we generated an immortalized monoclonal cell line with TPTC identities from giant pandas via lentiviral transduction of SV40 large T antigen into the cells and the subsequent clonal isolation through limiting dilution. The generated cell line, designated PD-TPTCs, demonstrated unlimited proliferative capacity and has been cultured for over five months and passaged more than 50 times to date. Characterization of PD-TPTCs revealed stable expression of key TPTC markers including ACTA2, MYH11, CNN1, and AR. Moreover, PD-TPTCs could respond to ATP and forskolin (FSK) stimulation with a pro-inflammatory gene expression profile and increased steroidogenic activity, respectively, and they were also amenable to lipofection. As such, the generated PD-TPTC line represents a promising cellular model for future mechanistic studies on the testicular niche, spermatogenesis, and reproductive disorders in giant pandas, laying the foundation for the development of novel assisted reproductive technology (ART) in this endangered species. Full article

This work reviews the complex role of the enzyme triosephosphate isomerase (TIM) (EC 5.3.1.1) within the context of diabetes, a prevalent metabolic disorder. It summarizes the main biochemical pathways, cellular mechanisms, and molecular interactions that highlight both the function of TIM and its implications in diabetes pathophysiology, particularly focusing on its regulatory role in glucose metabolism and insulin secretion. TIM’s involvement is detailed from its enzymatic action in glycolysis, influencing the equilibrium between dihydroxyacetone phosphate and glyceraldehyde-3-phosphate, to its broader implications in cellular metabolic processes. The article highlights how mutations in TIM can lead to metabolic inefficiencies that exacerbate diabetic conditions. It discusses the interaction of TIM with various cellular pathways, including its role in the ATP-sensitive potassium channels in pancreatic beta cells, which are crucial for insulin release. Moreover, we indicate the impact of oxidative stress in diabetes, noting how TIM is affected by reactive oxygen species, which can disrupt normal cellular functions and insulin signaling. The enzyme’s function is also tied to broader cellular and systemic processes, such as membrane fluidity and cellular signaling pathways, including the mammalian target of rapamycin, which are critical in the pathogenesis of diabetes and its complications. This review emphasizes the dual role of TIM in normal physiological and pathological states, suggesting that targeting TIM-related pathways could offer novel therapeutic strategies for managing diabetes. It encourages an integrated approach to understanding and treating diabetes, considering the multifaceted roles of biochemical players such as TIM that bridge metabolic, oxidative, and regulatory functions within the body. Full article