Search Results (8,322)

The phyllosphere, the aerial part of plants, serves as a crucial habitat for diverse microorganisms. Phyllosphere bacteria can activate protective mechanisms that help plants resist disease. This study focuses on isolating and characterizing phyllosphere bacteria from cucurbits to evaluate their potential in controlling Colletotrichum orbiculare, a pathogen causing anthracnose in cucumbers. Among the 76 bacterial isolates collected, 11 exhibited strong antagonistic effects against C. orbiculare in vitro. Morphological and 16S rRNA analyses identified these isolates as different Bacillus species, including B. vallismortis, B. velezensis, B. amyloliquefaciens, and B. subtilis. These bacteria demonstrated essential plant-growth-promoting and biocontrol traits, such as motility, biofilm formation, phosphate solubilization, nitrogen fixation, and the production of indole acetic acid. Most of the bacterial strains also produced biocontrol compounds such as ammonia, acetoin, siderophores, hydrogen cyanide, chitinase, protease, lipase, and cellulase. The application of these bacteria significantly enhanced cucumber growth in both non-manured and organically manured soils, showing improvements in root and shoot length, chlorophyll content, and biomass accumulation. Additionally, bacterial treatments effectively reduced anthracnose severity, with isolates GL-10 and L-1 showing the highest disease suppression in both soil types. Colonization studies showed that phyllobacteria preferentially colonized healthy leaves over roots and diseased tissues, and they were more effective in manure-amended soils. These results suggest that Bacillus phyllobacteria have strong potential as sustainable bio-stimulants and biocontrol agents, offering an effective approach for enhancing cucumber growth and disease control under both fertilized and unfertilized soil conditions. Full article

Background/Objectives: IBDs are chronic relapsing inflammatory intestinal disorders whose precise etiology is still only poorly defined: critical for their pathogenesis is the CCL20/CCR6 axis, whose modulation by small molecules may represent an innovative therapeutic approach. The aim of the present work is to test the potential efficacy of two molecules, MR120, a small selective CCR6 antagonist, active in TNBS- and chronic DSS-induced murine models of intestinal inflammation, and its derivative MR452, a well-tolerated agent endowed with improved anti-chemotactic in vitro properties, in the adoptive transfer colitis model. To the best of our knowledge, this is the first attempt to use adoptive transfer colitis to test modulators of the CCL20/CCR6 axis. Methods and Results: The induction of colitis in immunocompromised mice receiving CD4⁺CD25⁻ T cells i.p. resulted in a moderate inflammation and was met with limited protective responses following daily subcutaneous administration of MR120 or MR452 for 8 weeks. Both compounds significantly reduced colonic myeloperoxidase activity, and MR452 also lowered CCL20 levels in the gut, but they failed to prevent the increase in the Disease Activity Index, colon wall thickening, and macroscopic inflammation score. Conclusions: Our findings suggest that, despite the beneficial effects played by MR120 against subacute TNBS- and chronic DSS-induced colitis, the pharmacological targeting of the CCL20/CCR6 axis in the adoptive transfer model has a negligible effect in ameliorating the IBD-like phenotype driven by the altered intestinal immune homeostasis and by the disrupted function of immune-suppressive Treg cells. Full article

Alcohol Use Disorder (AUD) profoundly impacts individuals and society, driven by neurobiological adaptations that sustain chronicity and relapse. Emerging research highlights neuroinflammation, particularly microglial activation, as a central mechanism in AUD pathology. Ethanol engages microglia—the brain’s immune cells—through key signaling pathways such as Toll-like receptor 4 (TLR4) and the NLRP3 inflammasome, triggering the release of proinflammatory cytokines (IL-1β, TNF-α, IL-6). These mediators alter synaptic plasticity in addiction-related brain regions, including the ventral tegmental area, nucleus accumbens, amygdala, and lateral habenula, thereby exacerbating cravings, withdrawal symptoms, and relapse risk. Rodent models reveal that microglial priming disrupts dopamine signaling, heightening impulsivity and anxiety-like behaviors. Human studies corroborate these findings, demonstrating increased microglial activation markers in postmortem AUD brains and neuroimaging analyses. Notably, sex differences influence microglial reactivity, complicating AUD’s neuroimmune landscape and necessitating sex-specific research approaches. Microglia-targeted therapies—including minocycline, ibudilast, GLP-1 receptor agonists, and P2X7 receptor antagonists—promise to mitigate neuroinflammation and reduce alcohol intake, yet clinical validation remains limited. Addressing gaps such as biomarker identification, longitudinal human studies, and developmental mechanisms is critical. Leveraging multi-omics tools and advanced neuroimaging can refine microglia-based therapeutic strategies, offering innovative avenues to break the self-sustaining cycle of AUD. Full article

The in vitro granulosa cell (GC) model presents a valuable tool to explore antral follicle development. A full understanding of the reasons and blocking methods that occur during in vitro luteinization of sheep GCs, stimulated by serum culture, is a complex goal that has not been completely achieved. Herein, the phenomenon and causes of GC differentiation, as well as the methods for inhibiting luteinization in an in vitro culture system, were investigated by immunofluorescence, Western blot, RT-qPCR, and ELISA techniques. The results reveal that, when compared to fresh GCs, FSHR protein levels in primary GCs significantly decreased in serum-containing media, while STAR protein levels significantly increased, implying that sheep GCs can differentiate in serum-containing media. LH concentrations were significantly greater in serum-containing media compared to serum-free media. The LH receptor (LHR) mRNA expression in primary-generation GCs steadily increased with longer culture times, indicating that LH-LHR signaling leads to GC luteinization in vitro. In primary and second-generation GCs, 180 nmol/L BAY-899, an LHR-specific antagonist, significantly increased FSHR protein expression, reduced STAR protein synthesis, and inhibited P4 secretion within 48 h of in vitro culture compared to controls. BAY-899 showed no adverse effects on fifth-generation GCs growth, implying that BAY-899 can inhibit GC luteinization while not affecting cell proliferation. In conclusion, this study found that the LHR antagonist BAY-899 can preserve the features of sheep GCs in vitro by suppressing the spontaneous luteinization process caused by LH-LHR signaling, which has a key methodological implication for studying the mechanics of antral follicle formation in vivo. Full article

Background/Objectives: Estrogen deficiency-related menopause is associated with various physical and psychological symptoms. Although hormone replacement therapy (HRT) effectively alleviates these symptoms, its long-term use is associated with several side effects such as an increased risk of breast cancer and cardiovascular disease. Consequently, there is a growing interest in some plant-derived phytoestrogens that are considered safer alternatives to estrogen. Recent studies on Scrophularia buergeriana confirmed their anti-inflammatory and antioxidant properties; however, their effects on menopausal health remain unclear. Therefore, the aim of this study was to investigate the estrogen-like effects of S. buergeriana root (SB-R) extract, a potential phytoestrogen. Methods: Briefly, the MCF-7 cell line, a widely used in vitro model for assessing estrogen-like activity, was treated with SB-R extract and 17β-estradiol (E2; positive control) in the presence or absence of ICI 182,780 (Fulvestrant), an estrogen receptor antagonist. An E-screen assay and flow cytometry were performed to assess the effects of the treatments on cell proliferation and the cell cycle, respectively. Additionally, Western blotting and immunofluorescence assays were performed to elucidate the potential mechanisms underlying the estrogen-like effects of SB-R. Result: Treatment with SB-R extract promoted MCF-7 cell proliferation in a manner similar to E2. However, ICI 182,780 co-treatment inhibited the SB-R extract-induced increase in MCF-7 cell proliferation. Additionally, SB-R extract promoted cell cycle progression by increasing the proportion of cells in the S and G2/M phases. Moreover, Western blotting and immunofluorescence assays showed that SB-R extract increased the expression of estrogen receptor alpha (ERα). Furthermore, SB-R treatment activated downstream signaling pathways by enhancing AKT and ERK phosphorylation and upregulated the expression of cell cycle regulators, including cyclin D1, cyclin dependent kinase 4 (CDK4), cyclin E1, and cyclin dependent kinase 2 (CDK2). Conclusions: SB-R exhibits estrogen-like activity by activating ERα-mediated AKT and ERK pathways and thereby increasing the expression of proteins involved in cell cycle regulation. This makes it a promising phytoestrogen candidate and a safer alternative to conventional hormonal therapy for alleviating menopausal symptoms. Full article

Background/Objective: Orthodontic mini-implants (MI) create new niches that may alter the oral microbiota and modulate host immune responses. While clinical inflammation is not always evident, microbial and molecular changes may precede visible signs of peri-implant infection. This study investigated microbial shifts and inflammatory responses following MI placement, with a focus on Saccharibacteria, nitrate-reducing bacteria (NRB), and periodontopathogens. Methods: Saliva and peri mini-implant crevicular fluid (PMICF) samples were collected from eight orthodontic patients at baseline (T0), one week (T1), and one month (T2) after mini-implant placement. DNA was extracted from each saliva and PMICF sample and pooled across the eight patients for each time point. The pooled DNA were then subjected to 16S rRNA gene sequencing using the Oxford Nanopore MinION platform. Statistical analysis was performed to determine shifts in bacterial abundance, diversity, and co-occurrence patterns across the different sample types (saliva vs. PMICF) and time points. Results: Alpha diversity decreased in PMICF at T2, while it remained stable in saliva samples. Periodontopathogens (Porphyromonas gingivalis, Treponema denticola, Fusobacterium nucleatum) increased in PMICF at T2, while NRB and Saccharibacteria, along with a representative host bacterium (Schaalia odontolytica), remained relatively stable. Co-occurrence analysis showed antagonistic relationships between Saccahribacteria/NRB and periodontopathogens. IL-6 significantly decreased from T1 to T2, while CRP showed a non-significant downward trend. The expression of nitrate reductase genes narG and napA remained stable across time intervals. Conclusions: Despite no clinical inflammation, MI placement led to localized microbial shift and mild inflammatory responses. NRB and Saccharibacteria’s stability and antagonistic relationship to periodontopathogens may indicate that they could be involved in maintaining microbial homeostasis. These findings highlight possible early biomarkers and ecological strategies to support oral health in MI patients. Full article

Allopregnanolone (allo) and isoallopregnanolone (isoallo) are neuroactive steroid epimers that differ in hydroxyl orientation at carbon three. Allo is a potent GABA-A receptor agonist, while isoallo acts as an antagonist, influencing brain function through their interconversion. Their metabolism varies across brain regions due to enzyme distribution, with AKR1C1–AKR1C3 active in the brain and AKR1C4 restricted to the liver. In rats, AKR1C9 (liver) and AKR1C14 (intestine) perform similar roles. Beyond AKR1Cs, HSD17Bs regulate steroid balance, with HSD17B6 active in the liver, thyroid, and lung, while HSD17B10, a mitochondrial enzyme, influences metabolism in high-energy tissues. Our current data obtained using the GC-MS/MS platform show that allo and isoallo in rats undergo significant metabolic conversion, suggesting a regulatory role in neurosteroid action. High allo levels following isoallo injection indicate brain interconversion, while isoallo clears more slowly from blood and undergoes extensive conjugation. Metabolite patterns differ between brain and plasma—allo injection leads to 5α-DHP and isoallo production, whereas isoallo treatment primarily yields allo. Human plasma contains mostly sulfate/glucuronided steroids (2.4–6% non-sulfate/glucuronided), whereas male rats exhibit much higher free steroid levels (29–56%), likely due to the absence of zona reticularis. These findings highlight tissue-specific enzymatic differences, which may impact neurosteroid regulation and CNS disorders. Full article

Purinergic P2X7 receptors are involved in cellular processes such as inflammation, proliferation, and tissue remodeling, although their significance in human skin physiology remains poorly understood. In this study, we demonstrated strong P2X7 receptor immunoreactivity in the basal and granular layers of the epidermis. Cutaneous expression of P2X7 receptors was further confirmed at the level of specific mRNA and protein in cultured primary human keratinocytes and dermal fibroblasts. To reveal a possible role of these receptors in regulation of keratinocyte and fibroblast function, the cells were treated with a P2X7 agonist BzATP, or its selective antagonist A438079. Cell proliferation and viability were assessed using an immunofluorescence-based cell counter, and cell migration was evaluated by wound healing assay. P2X7 stimulation with BzATP significantly inhibited keratinocyte proliferation and migration, while P2X7 inhibition with A438079 significantly enhanced keratinocyte migration. In contrast, fibroblasts displayed minimal response to either treatment. These findings indicate that P2X7 regulates keratinocyte growth, and purinergic signaling may play a role in the skin. Our data also suggest that selective P2X7 inhibition may support re-epithelialization under conditions associated with impaired wound healing. Full article

The utilization of high-volume fly ash (HVFA, ≥50% cement replacement) in concrete is pivotal for sustainable construction but hindered by low early-age strength. This study investigates the individual and combined effects of hydrated lime (HL) and calcium formate (CF) on the strength development, hydration kinetics, and microstructure of HVFA pastes (60% and 70% FA). Individual additions of 11% HL (HVFA60) or 14% HL (HVFA70) raised 28-day compressive strength by 18% and 22%, respectively, and shortened final setting from 10.0 h to 3.8 h. Similarly, 3% CF increased 28-day strength by 15% (HVFA60) and 12% (HVFA70) while cutting final setting to 2.1 h and 3.3 h. In contrast, combining HL and CF suppressed strength by 15–22% despite accelerating final setting to less than 1 h. Isothermal calorimetry showed a 40% reduction in cumulative heat release at 44 h for the combined system. XRD, TGA and SEM confirmed 20–30% lower C-S-H content, 25% less CH, and a rise in porosity when HL and CF were used together. These findings demonstrate that HL and CF act as competing accelerators, where rapid heat release compromises microstructural integrity. For practical applications using HVFA materials, individual use of HL or CF is recommended to enhance early-age performance, while combined application should be avoided to prevent strength reduction. Full article

This investigation on novel antifungal agents featuring a thiol-reactive (dithioperoxo)thiolate chemical nucleus [-NC(S)S-SR] established that the optimal levels of fungal growth inhibition were achieved with thiomethyl-bound derivatives (R = Me). The most efficacious analogs had MIC₅₀/MIC₉₀ values of 2/2 µg/mL and an MIC range of 1 to 2 µg/mL for a ten-member panel of voriconazole-resistant A. fumigatus mutants. Pharmacodynamic studies revealed that the lead (dithioperoxo)thiolates impaired conidial germination and germling development more effectively than voriconazole for the triazole-resistant strain AR-1295. Moreover, glutathione and Cu²⁺ were shown to have antagonistic interactions, which was attributed to the thiol-reactive, pro-oxidant properties of the (dithioperoxo)thiolates and their metabolic conversion to chelating agents. Cytotoxicity studies further showed that the compounds were less toxic to human fetal kidney cells than squamous carcinoma cells. The collective findings of the investigation indicate that (dithioperoxo)thiolates are effective antifungal agents against A. fumigatus to merit additional research on their therapeutic potential. Full article

Neuroendocrine tumors (NETs) are a rare and heterogeneous class of neoplastic lesions, but their prevalence has increased significantly over the past three decades. These tumors are aggressive and difficult to treat. Improving diagnostic efficiency and treatment effectiveness is important for patients with neuroendocrine tumors. Radiopharmaceutical therapeutic diagnostics combines diagnosis and treatment technology and has broad prospects in precision medicine, especially for the early diagnosis and treatment of tumors. To compare the diagnostic advantages of radiolabeled somatostatin receptor agonists and antagonists for liver metastases from NETs and the disease control rate in NET patients. Systematic search of PubMed, Embase, Cochrane, Ovid, Scopus, and Web of Science databases up to 29 October 2024. Clinical trials of somatostatin receptor agonists and antagonists for NET diagnosis or treatment. Following PRISMA guidelines, data were independently extracted by two researchers. Pooled diagnostic or treatment effects and 95% CIs were reported using a random-effects meta-analysis model. Effect of somatostatin receptor agonists and antagonists in detecting liver metastases and disease control rate. Risk Ratio (RR) for liver metastasis detection and Effect Size (ES) for disease control rate were calculated. From 5291 articles, 52 were included in the meta-analysis. Radiolabeled somatostatin receptor antagonists were significantly more effective than agonists in detecting liver lesions (RR = 11.57, 95% CI: 4.10, 32.67). Disease control rates were higher with antagonists (ES = 0.90, 95% CI: 0.83, 0.96) compared to agonists (ES = 0.82, 95% CI: 0.78, 0.85, z = 2.12, p = 0.03). Radiolabeled somatostatin receptor antagonists outperform agonists in diagnosing hepatic lesions and controlling disease in NETs, highlighting their clinical superiority. This meta-analysis provides critical insights into the diagnostic and therapeutic efficacy of somatostatin receptor antagonists, and may offer a potential paradigm shift in the management of neuroendocrine tumors. Nevertheless, the smaller number of studies on antagonists may limit the generalizability of the findings and underscore the need for further clinical trials to validate these results. Full article

Gulf War illness (GWI) is a multi-symptom disorder affecting veterans of the Persian Gulf operations. Persistent neuroendocrine dysregulation contributes to impairing cognitive capacity and generates anxiety-like behavior. Effective treatments for this illness are challenging due to compromised metabolism, increased oxidative stress and neuroinflammation, perpetuated by chronic stress and hypothalamic–pituitary–adrenal (HPA) axis dysfunction. This neuroinflammation can be alleviated with synthetic antagonistic analogs of the growth hormone-releasing hormone (GHRH) through modulation of the HPA axis. We evaluated the efficacy of the GHRH antagonist analog, MIA-690, against cognitive impairment and anxiety-like behavior in GWI. Mice exposed to an experimental GWI model involving corticosterone (CORT) and diisopropylfluorophosphate (DFP), followed by CORT and lipopolysaccharide (LPS), received a daily subcutaneous dose of 10 μg of MIA-690 for 10 days. Assessments of spatial memory, recognition capacity, somatic health, anxiety and innate survival were carried out, combining the Morris water maze (MWM), novel object recognition (NORT), grip strength (GST), and open field (OFT) tests. Learning efficiency was selectively enhanced in females using the MWM. There were no significant differences in the recall capacity and performance on the OFT, NOR, and GST tasks. Our findings suggest that the MIA-690 dosage is sufficient to improve learning deficits in experimental GWI exposures. Full article

Amphibian populations have experienced a severe decline over the past 40 years, driven primarily by environmental pollution, habitat destruction, climate change, and disease. This work reports, for the first time, saprolegniosis in Pelophylax perezi egg masses and saprolegniosis in amphibians in Portugal. After isolation and phylogenetic analysis, the pathogen was identified as Saprolegnia australis. Following this, the present work intended to screen a collection of P. perezi skin bacteria for the existence of bacterial strains with inhibitory action against the newly identified S. australis SC1 and two other species, Saprolegnia diclina SAP 1010 UE and Saprolegnia australis SAP 1581 UE. The results showed that various bacterial species could inhibit the growth of these three species of oomycetes. Bacteria with the most significant antagonistic action against Saprolegnia spp. predominantly belonged to the genus Bacillus, followed by Serratia, Pseudomonas, and Aeromonas. Despite variations in bacterial diversity among frog populations, the present study also demonstrated the presence of bacteria on frogs’ skin that were capable of inhibiting Saprolegnia spp., as evidenced by in vitro challenge assays. These findings highlight the protective function of bacteria present in amphibian skin. The observed bacterial diversity may contribute to the metabolic redundancy of the frog skin microbiome, helping to maintain its functional capacity despite shifts in the community composition. Additionally, the study found that, when providing a more advantageous environment for pathogen growth—in this case a peptone–glucose (PG) medium instead of R2A—the percentage of bacteria with moderate-to-strong antagonistic activity dropped by 13% to 4%. In conclusion, the presence of bacteria capable of inhibiting Saprolegnia spp. in adult individuals and across different environmental conditions may contribute to lowering the susceptibility of frog adults towards Saprolegnia spp., compared with that in the early stages of development, like the tadpole or egg stages. Full article

Signed networks, which incorporate both cooperative and antagonistic interactions, naturally give rise to symmetric behaviors in multi-agent systems. One such behavior is bipartite containment tracking, where follower agents converge to a symmetric configuration determined by multiple groups of leaders with opposing influence. Moreover, a timely response is critical to ensuring high performance in containment tracking tasks, particularly for high-order multi-agent systems operating in dynamic and uncertain environments. To this end, this paper investigates the predefined-time bipartite containment tracking problem for high-order multi-agent systems affected by external disturbances. A robust tracking control scheme is developed based on the backstepping method to ensure that the tracking errors converge to a predefined residual set within a user-specified time. The convergence time is explicitly adjustable through a design parameter, and the proposed scheme effectively avoids the singularities often encountered in conventional predefined-time control approaches. The stability and robustness of the proposed scheme are rigorously established through Lyapunov-based analysis, and extensive simulation results are provided to validate our theoretical findings. Full article

The corticotropin-releasing factor (CRF) and its type 1 receptor (CRF₁R) play a key role in the regulation of the hypothalamic–pituitary–adrenal (HPA) axis. Dysregulation of the HPA axis is associated with congenital adrenal hyperplasia (CAH) and depression. Non-peptide CRF₁R-selective antagonists displayed antidepressant effects on animal models and are used for the management of CAH. To develop novel non-peptide CRF₁R antagonists, we have previously designed and synthesized a series of substituted pyrimidines. Among these analogs, molecule 43 (M43) binds to CRF₁R with the highest affinity. Based on this finding, we selected M43 for further pharmacological characterization in the present study. The results suggest that M43 is a potent CRF₁R antagonist, blocking the ability of the CRF-related agonist, Tyr⁰-sauvagine, to stimulate (1) cAMP accumulation in HEK 293 cells expressing CRF₁R and (2) the proliferation rate of RAW 264.7 macrophages. Computational studies suggest that the antagonist properties of M43 are mostly attributed to its ability to interact with residues in the allosteric pocket of CRF₁R, comprised of the third, fifth, and sixth transmembrane domain residues, which block activation-associated structural rearrangements of the receptor. Our data will be used to design novel non-peptide CRF₁R antagonists for clinical use. Full article