Polydopamine-functionalized sodium alginate nanoparticles (SA-PDAs) were fabricated through self-assembly of dopamine-modified sodium alginate with cetyltrimethylammonium bromide and Tween 80, followed by in situ oxidative polymerization. The nanoparticle served as an excellent carrier for the controlled release of emamectin benzoate (EB) with enhanced photostability and foliar adhesion. The release of EB from nanoparticles, dominated by interactions between the active ingredient and the matrix, was sensitive to the pH of the medium, near-infrared irradiation, and protease hydrolysis. The encapsulation to nanoparticles significantly improved the insecticidal efficacy of EB while alleviating acute toxicity to the growth of cucumber seeds and the survival of earthworms and zebrafish. The median lethal concentration of the EB@SA-PDA nanosuspension against Mythimna separata larvae was only 34.7% of that of the commercial EB emulsifiable concentrate. The results illustrated a straightforward and effective strategy for constructing an intelligent platform to translate nanopesticides from laboratory investigations into field applications.

Bcl-2-associated athanogene 3 (BAG3) is a mediator of chaperone-assisted selective autophagy, and in the brain, is most highly expressed in astrocytes. However, its role in astrocytes remains poorly defined. Given the genetic and pathological links of BAG3 to proteostasis and neurodegenerative diseases, we investigated how BAG3 contributes to astrocyte function and Alzheimer's disease (AD). To define its function and relevance, we used single-nucleus RNA sequencing to confirm BAG3 enrichment in astrocytes and employed CRISPR/Cas9 editing of human induced pluripotent stem cells followed by proteomic and transcriptomic profiling, which revealed that BAG3 loss caused greater disruption in astrocytes than in neurons. BAG3-deficient astrocytes displayed reduced autophagy, lysosomal abundance and activity, and proteasome function. Coimmunoprecipitation identified BAG3 known binding partners (e.g., HSPB8, proteasome regulators), as well as an interactor in the retromer complex, VPS35. BAG3 deficiency resulted in altered retromer activity as measured by amyloid precursor protein (APP) localization in endosomes. In addition to validating these binding partners, integrative -omics analyses showed that BAG3 regulates AD-relevant proteins (GFAP, BIN1), as well as HSPB8. Functionally, BAG3 knockout astrocytes exhibited impaired amyloid-β proteostasis when cocultured with APP/PSEN1 mutant neurons, directly linking BAG3 to a disease-relevant astrocyte phenotype. Finally, analysis of postmortem human brain revealed that BAG3 expression marks a stress-responsive astrocyte subtype in aged individuals. Together, these findings demonstrate that BAG3 coordinates astrocyte proteostasis through interactions with regulators of autophagy, proteasome activity, and retromer function, positioning it as a potential therapeutic target and central node of astrocytic protein quality control in neurodegeneration.

Acute myeloid leukemia (AML) critically depends on oxidative phosphorylation (OXPHOS). Mitochondrial targeted therapies are advancing into clinical trials, but metabolic vulnerabilities have been primarily explored from a tumor-intrinsic perspective. Leukemic cells can actively reshape an immunosuppressive tumor microenvironment through metabolic competition, highlighting the importance of evaluating the impact of mitochondrial targeted therapies on antitumor immunity. Here, we showed that elevated expression of the mitochondrial caseinolytic protease P (ClpP) is associated with advanced AML. Development of IMP125, a potent ClpP agonist, enabled dual targeting of AML to elicit robust anti-leukemic activity. In addition to direct cytotoxicity, IMP125 induced immunometabolic reprogramming of AML models. Owing to differential ClpP expression, IMP125 preferentially suppressed mitochondrial respiration and oxygen consumption in AML cells while exerting minimal direct effects on T-cell respiration. By reducing AML oxygen consumption and metabolic demand, IMP125 alleviated hypoxia and metabolic competition within the leukemic niche. In this remodeled microenvironment, T cells exhibited recovery of OXPHOS, effector function, and memory-associated features, consistent with an indirect mechanism mediated through metabolic suppression in AML cells. The anti-leukemic efficacy of IMP125 was dependent on T cells and was synergistically enhanced by PD-1 blockade. Collectively, this work reframes ClpP agonism from a tumor-intrinsic therapy to a strategy that actively induces metabolic-immune rewiring, bridging ClpP-targeted and immunotherapeutic approaches for mitochondrial-dependent AML.

Understanding the developmental dynamics of immune system is essential for improving larval health management. This study examined the ontogenetic patterns of seven innate immune-associated parameters, viz. albumin, protease, antiprotease, sialic acid, nitrite, nitric oxide synthase (NOS), and alkaline phosphatase (ALP) along with a general clinical indicator of health (total protein content) from fertilized eggs (0 DPH) to the completion of metamorphosis (25 DPH) in a high-value tropical mariculture species, Trachinotus blochii. Among all parameters, protein content showed the least inter-stage variation (26.40%). All immune-associated markers displayed distinct, non-linear trajectories rather than a gradual, uniform progression. Larval development was marked by a steady rise in protease and NOS activities along with increasing nitrite levels. Metamorphosis was accompanied by marked changes in several immune-associated parameters, characterized by significant increases in albumin, nitrite, protease, and NOS, and simultaneous decreases in ALP, antiprotease, and sialic acid. These coordinated changes suggest intense modulation of innate immunity during the larva-to-juvenile shift. Post-metamorphic stages showed stabilization of several immune-associated markers, indicating the establishment of regulatory homeostasis. Multivariate analysis grouped early ontogeny into three clusters, with the greatest divergence observed between eggs and developmental stages undergoing metamorphosis. The present study also provides preliminary baseline reference values for innate immune-associated parameters during larval development of T. blochii. These findings contribute to the understanding of immune ontogeny in this species and may serve as a reference for future studies investigating larval immune development and hatchery management strategies following validation across multiple batches and functional immune assessments.

The increasing emergence of drug-resistant fungal pathogens has heightened the search for new antimicrobial compounds with previously unexplored modes of action. Among candidate molecules for control of fungal diseases, plant antimicrobial peptides (AMPs) have attracted considerable interest because they are natural molecules with broad-spectrum inhibitory activities against microorganisms. In this study, peptide-enriched fractions obtained from Capsicum frutescens seeds were evaluated for antimicrobial activity and mechanisms of action against Candida species. Hemotoxicity was evaluated in vitro using sheep erythrocyte lysis, and inhibitory effects against serine protease and α-amylase enzymes were also determined. Initially, the protein-rich extract obtained was fractionated by reverse-phase chromatography on an HPLC system, resulting in five major fractions (F1-F5). Enzymatic assay revealed that all these fractions inhibited the activity of human salivary α-amylase, with F2 exhibiting 100% inhibition of activity. Fractions F2 and F3 exhibited the highest antifungal activity, inhibiting the growth of Candida albicans by 83% and 69% and Candida parapsilosis by 71% and 57%, respectively (200 µg mL- 1). These fractions reduced cell viability and increased membrane permeabilization. Additional mechanistic analyses suggested distinct cellular effects, with F2 associated with alterations in cell wall integrity and F3 associated with mitochondrial dysfunction. Partial sequence analysis indicated similarity to plant defense peptides, including heveins, defensins, and vicilin proteins. Importantly, both fractions exhibited low hemolytic activity, indicating selectivity toward fungal cells. Thus, the results highlight the potential of peptide-enriched fractions from C. frutescens seeds as multifunctional antifungal agents and support their relevance for the development of new strategies to control Candida infections.

Endometriosis is a common gynecological disease in which lipid metabolic dysregulation has been increasingly implicated. However, the specific metabolic changes in ectopic lesions and stromal cells remain poorly understood. It is also unclear whether these changes contribute to lipid droplet accumulation or how this process is controlled at the transcriptional level. Here, we report that ectopic stromal cells exhibit broad lipid remodeling, showing changes in phosphatidylcholines, lysophosphatidylcholines, sphingomyelins, and acylcarnitines. Both ectopic lesions and ectopic stromal cells showed increased triacylglycerol and total cholesterol levels, lipid droplet accumulation, and reduced E2F1 expression. Integrated epigenomic and transcriptomic analyses of patient-derived ectopic stromal cells, together with mechanistic validation, revealed that E2F1 transcriptionally activates MGLL, which encodes monoacylglycerol lipase, a lipolytic enzyme involved in lipid mobilization. Both in vitro and in vivo experiments showed that downregulation of E2F1 increased lipid droplet accumulation, whereas restoring MGLL expression partially reversed this effect. Consistently, targeted lipidomic analysis of ectopic stromal cells following MGLL knockdown showed increased levels of several diacylglycerol species and alterations in sphingomyelin profiles. In summary, our findings suggest that E2F1 downregulation reduces MGLL expression and impairs lipid mobilization, which may contribute to lipid droplet accumulation and endometriotic lesion growth.

Ubiquitin-specific proteases (USPs) modulate diverse virus-host interaction events through deubiquitination; thus, investigating their functional roles and molecular targets facilitates the interpretation of viral pathogenic mechanisms and informs the design of anti-virus strategies. The deubiquitinase USP8 has been reported to antagonize HIV-1 replication by reversing Vif-mediated ubiquitination of the host restriction factor A3G and shielding it from proteasomal degradation. Here, we identify an additional A3G-independent anti-HIV mechanism of USP8. In A3G-negative cells, USP8 exerts a potent inhibitory effect on HIV-1 replication by significantly suppressing viral transcription. Mechanistically, USP8 interacts with the HIV-1 trans-activator Tat through its C-terminal domain and mediates the removal of Tat's polyubiquitination chains in a manner dependent on USP8 catalytic activity. This disrupts the association between Tat and the HIV-1 LTR transactivation response (TAR) element, thus impairing Tat-mediated viral transactivation. Furthermore, given the essential role of Tat in driving viral gene expression, silencing of USP8 triggers latent HIV-1 reactivation. Collectively, our results, along with previous work, reveal that USP8 acts as a compelling anti-HIV factor that, by regulating both host and viral proteins, ultimately suppresses HIV-1 replication. These results deepen our understanding of USP8-mediated HIV-1 restriction and provide insights for the development of novel anti-HIV therapeutic strategies.IMPORTANCEUbiquitination modification contributes a non-negligible part to the regulation of HIV-1 replication, and the interpretation of key factors involved in this process has significantly benefited our understanding of the virus-host interplay. USP8 is a ubiquitin-specific protease that was recently identified to restrict HIV-1 replication by stabilizing the host restriction factor A3G through reversing the Vif-mediated ubiquitination of A3G. Here, we demonstrate an additional A3G-independent anti-HIV mechanism of USP8. In A3G-negative cells, USP8 directly interacts with and deubiquitinates Tat to cleave its K63-linked ubiquitination, thereby suppressing Tat-mediated HIV-1 trans-activation. The suppression of Tat-driven viral transcription induced by USP8 requires the enzymatic activity of USP8 and is effective in different HIV-1 tropic strains. In addition, the fact that Tat is a novel substrate of USP8 is validated both in cells and by in vitro deubiquitination assays. These findings illustrate the dual regulatory roles of USP8 on both host antiviral factors and viral proteins, expanding our understanding of host-mediated HIV-1 restriction.

Cysteine proteases are recognized as relevant therapeutic targets in both parasitic diseases and cancer due to their essential biological functions. Among these enzymes, cruzain from Trypanosoma cruzi is an important molecular target for the treatment of Chagas disease. Likewise, human cathepsin L (hCatL) is a potential target for anticancer drug development. Both enzymes share conserved catalytic features, particularly the cysteine-histidine catalytic dyad, which enables interaction with nitrile-based reversible covalent inhibitors. Here, six nitrile-derived inhibitors (Neq1171.1, Neq1172.1, Neq1181.1, Neq1182.1, Neq1268, and Neq1276) had their cruzain and hCatL inhibition constants, Ki, determined. Besides, we have used molecular dynamics simulation and alchemical free energy calculations for evaluating the overall relative binding free energies of these inhibitors. The calculated free energy differences closely reproduced experimental trends, confirming the reliability of the computational protocol. These findings demonstrate that thermodynamic integration is an effective strategy for predicting relative binding affinities and provides valuable molecular-level insights for the rational optimization of cysteine protease inhibitors with potential applications in Chagas disease and cancer therapy.

This article critically examines ADAM19-mediated inflammatory remodeling within five distinct subtypes of solid tumors prone to inflammation, employing non-neoplastic inflammatory states strictly as a comparative reference. As a transmembrane protease belonging to the ADAM family, ADAM19 undergoes maturation through plasmin-dependent cleavage, with its expression tightly regulated by both transcriptional and epigenetic controls. Although dysregulated ADAM19 levels are evident in various inflammatory lesions and malignancies, its context-specific dual biological roles necessitate a systematic synthesis.

The storage of root vegetables is a critical aspect of agricultural production, significantly impacting both the quality and quantity of produce available for consumption. This study aimed to develop a bacterial consortium capable of preventing storage bacterial diseases in root vegetables. A total of 225 bacterial strains were isolated from various ecological niches, including crops, peri-root soils, coastal dune soils, sewage sludge, and symbiotic bacteria of common insect pests. These strains were characterized for their ability to grow at storage temperatures (4 °C) and produce cellulase, lipase, protease, and chitinase enzymes. From 225 isolates, 24 strains exhibited the highest enzymatic activities were selected for mutual inhibition assays, resulting in the selection of four strains (SP65, Dv004, Dv008, and Dv024a) that demonstrated high growth rates and enzymatic activities. The selected strains grew at 4 °C (OD600 range: 0.061-0,190) and they exhibited the high intensity of production of the following enzymes measured as a halo diameter (cm) on bacterial medium: cellulase (from 1,4 to 1,8 cm), lipase (0,7) - 1,5), proteinase 1,2 - 1,6) and chitinase (0,3 - 0,4). These strains, identified as Stenotrophomonas maltophilia (SP65) and Serratia liquefaciens, were combined to form a bacterial consortium. The efficacy of this consortium was tested on potato (Solanum tuberosum L.), carrot (Daucus carota L.), celeriac (Apium graveolens L.), parsley (Petroselinum crispum Mill.), and onion (Allium cepa L.). The mixed culture was tested against bacterial soft rot pathogens including Pectobacterium carotovorum, Kosakonia cowanii, Enterobacter cloacae subsp. dissolvens and Burkholderia cepacia. For celeriac and parsley, the effect of inhibition of Pectobacterium carotovorum, Burkholderia cepacia, E. cloaceae subsp. dissolvens, Kosakonia cowanii, and the mixture of those pathogens were observed when pathogens were applied at concentration of 1,3 × 10[8] CFU mL[-1]. In case of onion the same effect of inhibition of growth of pathogens were observed only for P. carotovorum, and for B. cepacia at concentration of 1,3 × 10[8] CFU mL[-1]. The results demonstrated the potential of using a bacterial consortium as a biological control method to enhance the storage life and quality of root vegetables.