Ovarian granulosa cells (GCs) are central to follicular development in goats, and their proliferation is tightly regulated by estrogen, yet the downstream signaling cascade mediating this effect in ruminants remains undefined. This study aimed to investigate whether estrogen regulates goat ovarian GC proliferation through the nitric oxide (NO) signaling pathway and to further elucidate the molecular mechanisms by which NO modulates GC proliferation. Primary goat GCs were treated with 17β-estradiol (17β-E2), the NO donor sodium nitroprusside (SNP), the NOS inhibitor NG-monomethyl-L-arginine (L-NMMA), the Wnt inhibitor FH535, and the Wnt agonist SKL2001. Cell viability, proliferation, and protein expression were assessed by CCK‑8, EdU, and Western blotting. 17β-E2 at concentrations of 1, 2, and 10 μg/mL all promoted GC viability (P < 0.05), with maximal effect at 1 μg/mL (P < 0.01), and simultaneously upregulated inducible NOS (iNOS), endothelial NOS (eNOS), and neuronal NOS (nNOS) protein expression via estrogen receptor α (ERα). The NO donor SNP enhanced GC proliferation, whereas the NOS inhibitor L‑NMMA suppressed it; co‑treatment with SNP partially rescued the L‑NMMA‑induced inhibition. Mechanistically, SNP increased β‑catenin and proliferating cell nuclear antigen (PCNA) protein levels, an effect reversed by FH535, whereas L‑NMMA‑induced NO depletion reduced their expression, which was restored by SKL2001, positioning NO upstream of the Wnt/β‑catenin pathway. Collectively, these findings establish that 17β-E2 promotes goat GC proliferation through a signaling cascade involving ERα, NOS, NO, and the Wnt/β‑catenin pathway, providing the functional evidence for this axis in a ruminant model and offering a mechanistic framework for understanding ovarian follicular dynamics with implications for improving reproductive efficiency in goats.

We synthesised a series of N[2],N[4]-diaminoquinazoline derivatives, among which 3-[[4-(benzylamino)quinazolin-2-yl]amino]benzenesulfonamide (compound 5) exhibited potent phosphodiesterase type 5 (PDE5) inhibitory activity. Increasing evidence suggests that PDE5 inhibition is a promising therapeutic strategy for peripheral vascular disease. This study aimed to investigate vasorelaxant effects of 5 and its underlying mechanisms in rat mesenteric arteries. Compound 5 was synthesised and purified, and evaluated its vascular activity using organ bath assay. Molecular docking was performed to identify its potential molecular targets. Compound 5 (0.0001-30 μM) induced relaxation of phenylephrine (PE)-precontracted mesenteric arteries (EC50 = 0.76 ± 0.29 μM; Emax = 95.6% ± 1.1%). This effect was attenuated by endothelium removal (EC50 = 5.24 ± 0.98 μM) and by L-NAME (eNOS inhibitor). Compound 5 enhanced sodium nitroprusside-induced relaxation, supporting modulation of the NO-sGC-cGMP axis via PDE5 inhibition. Iberiotoxin (BKCa blocker) reduced relaxation, whereas 4-aminopyridine (KV blocker) and glibenclamide (KATP blocker) had no effect. Compound 5 inhibited vasoconstriction mediated by extracellular Ca[2+] influx via receptor-operated Ca[2+] channels (ROCCs) and α1-adrenergic receptor activation. Molecular docking provided qualitative support for PDE5A, BKCa channels and α1-adrenergic receptor as potential targets. In conclusion, 5 induces potent vasorelaxation in resistance arteries through eNOS/NO/cGMP signalling, BKCa channel activation and inhibition of ROCCs and α1-adrenergic receptors, supporting its potential as a therapeutic candidate for peripheral vascular disorders.

Copper-based nanomedicines have broader applications in the biomedical sector because of their unique fundamental properties. Very recently, our group has designed and synthesized new copper nitroprusside analogue nanoparticles (CuNPANP in vitro and in vivo) anticancer therapy in both breast cancer and melanoma. The toxicity study of these therapeutically active nanomedicines in small and large animals is a prerequisite before their clinical translation. In the present manuscript, detailed toxicity studies of CuNPANP are demonstrated through various in vitro and in vivo studies. Initially, the biocompatible nature of the CuNPANP has been established through several in vitro assays (cell viability assay, cell cycle study, apoptosis analysis, and determination of reactive oxygen species using DCFDA) in RAW 264.7 cells. Further, the noninflammatory nature of the nanoparticles toward RAW 264.7 cells has been investigated through immunocytochemistry (COX-2 staining) and Western blot analysis. The in vivo toxicity studies of CuNPANP have been performed in a BALB/c mouse (male and female) model using therapeutic doses (1 and 5 mg kg-1 body weight (b.w) in a time-dependent manner (30, 60, and 120 days). There were no significant changes observed in different body parameters (body weight, feed consumption, and relative organ weight) of the mice and no major alterations were found in blood hematology and serum biochemical measurements, while we compare the CuNPANP-treated groups with the UT group. Furthermore, the histopathological examination of male mice group upon 30, 60, and 120 days exposure reveals the biocompatibility of CuNPANP. Further, the biodistribution study reveals the distribution of copper in vital organs. Altogether, the results demonstrate the nontoxicity and biocompatibility of the nanomaterials.

Despite the great potential of direct seawater electrolysis technology in alleviating freshwater scarcity for green hydrogen production, its practical application is severely limited by the degradation of anode catalysts under chloride‑rich conditions. In this study, we report a sodium nitroprusside‑modified NiFe layered double hydroxide on nickel foam (SNP@NiFe LDH/NF) as an anode for alkaline seawater oxidation electrocatalysis. Such SNP@NiFe LDH/NF delivers a current density (j) of 1000 mA cm[-2] at an overpotential of 380 mV. Meanwhile, it demonstrates excellent long‑term durability, sustaining operation for 1100 h at 1000 mA cm[-2] and 900 h at 2000 mA cm[-2], with a total operating time of 2000 h. Furthermore, the anion-exchange membrane electrolyzer constructed using SNP@NiFe LDH/NF and Pt/C/NF as the anode and cathode, respectively, delivers a j of 500 mA cm[-2] at a cell voltage of 2.30 V, while running stably for 600 h.

It is known that the progressive climate changes and environmental pollution prevalent all over the world lead to increasingly frequent and prolonged episodes of stress, and it challenges plants to adapt to changing adverse conditions. Given the significant crop losses caused by challenging environments, protecting plants from the destructive effects of these factors has become one of the most pressing challenges in modern agricultural biology and plant physiology. One of the approaches to protect plants from stressors in modern agrobiotechnology is the use of nitric oxide (NO) donors, which are chemicals that can release NO in plant cells and stimulate their germination, growth, biomass accumulation, and protection against external factors. Studying their mechanism of action at the molecular level is essential for understanding the targets of NO action, as well as for developing new phytoeffectors. A multiomics approach, along with physiological data, is often the method of choice. In this review, we compare widely used NO donors (e.g., sodium nitroprusside, S-nitrosoglutathione, NONOates) in terms of chemical properties, NO-release kinetics, stability, and suitability for different application methods (seed treatment, foliar spray, root application). Finally, we highlight recent advances from proteomics, metabolomics, and transcriptomics that have enabled system-wide identification of NO-responsive pathways and candidate targets. By integrating physiological, biochemical, and multiomics data, this review provides a framework for understanding NO-mediated stress tolerance and for the rational design of new NO-based phytoeffectors.

Pseudomonas aeruginosa is a prominent biofilm-forming pathogen associated with chronic infections that are difficult to eradicate due to the protective biofilm matrix. Inducing biofilm dispersion has emerged as a potential therapeutic strategy. Although researchers have compared selected biofilm dispersion mechanisms, the characteristics of cells generated by mechanistically distinct release strategies, particularly enzymatic matrix degradation, remain uncharacterized. In this study, we compared P. aeruginosa cells released from biofilms using four distinct dispersion mechanisms: An external signaling molecule (cis-2-decenoic acid; cis-DA), a nutrient cue (glutamate), an environmental toxin (sodium nitroprusside; SNP), and a matrix-degrading enzyme (amylase). Released cells exhibited distinct antibiotic susceptibility, attachment, and gene expression depending on the release mechanism. Glutamate-released cells exhibited greater tobramycin tolerance and attachment, whereas glutamate and amylase-released populations exhibited differences in the expression of selected genes. Additionally, the release mechanism influenced subsequent biofilm recovery and reformation, and cis-DA significantly enhanced tobramycin-mediated killing of mature biofilms. Collectively, these findings demonstrate that the mechanism used to induce release may influence subsequent bacterial behavior and antibiotic susceptibility. These findings highlight the importance of considering release mechanism when developing dispersion-based strategies for treating biofilm-associated infections.

High-temperature stress during flowering poses a severe threat to global rice production. Although nitric oxide (NO) is known to mediate plant stress responses, its protective role during this critical developmental stage remains poorly understood. In this study, we investigated the protective effects of the NO donor sodium nitroprusside (SNP) on a heat-tolerant cultivar (N22) and a heat-sensitive cultivar (YR343) exposed to a dynamic high-temperature regime (comprising five gradual transitions over a 24-h cycle within 30-38 °C). The results showed that foliar application of SNP at 150 μmol L[-1], the most effective concentration among those tested in this study, significantly alleviated high-temperature-induced yield losses in both cultivars, with a notable genotype-dependent effect. In the heat-sensitive YR343, yield recovered by 74.3%, compared with only 35.8% in the heat-tolerant N22. This differential responsiveness was associated with a coordinated multi-physiological network. SNP promoted transpirational cooling, lowering panicle and leaf temperatures by up to 1.87 °C, preserved photosynthetic performance, and activated antioxidant enzymes (SOD, POD, and CAT) to alleviate oxidative membrane damage. Concurrently, SNP sustained nitrogen assimilation (via NR, GS, GOGAT, and GDH) and osmolyte accumulation, while rebalancing endogenous hormone profiles by elevating IAA and CTK levels and suppressing ABA accumulation. Notably, these protective effects were more pronounced in the sensitive genotype, suggesting that exogenous NO compensates for inherent physiological weaknesses in heat-susceptible cultivars. Collectively, these findings support the use of SNP as an effective strategy for improving rice thermotolerance at flowering, particularly for stabilizing yield in heat-susceptible genotypes. Moreover, this work provides physiological insights consistent with a proposed NO-coordinated stress mitigation and presents a genotype-sensitive strategy for sustaining rice productivity under a warming climate.

Although zinc (Zn) deficiency severely inhibits rice growth, it remains unclear how melatonin (MT) promotes adaptation under conditions of insufficient Zn supply. To address this issue, rice seedlings were treated with MT, the NO donor sodium nitroprusside (SNP), or the NO scavenger c-PTIO, alone or in combination, under Zn-deficient conditions. Zn starvation rapidly promoted endogenous MT accumulation in rice roots, together with a 90.2%-179.5% rise in the transcript levels of MT biosynthetic genes. Exposure to Zn deprivation reduced root/shoot length and root/shoot biomass, by 46.2%/31.3% and 27.5%/39.6%, respectively. Among the tested concentrations, 10 μM MT produced the strongest protective effect, increasing root length, shoot height, root biomass, and shoot biomass by 59.9%, 34.2%, 21.7%, and 51.8%, respectively, relative to plants that were merely deficient in Zn. In Zn-deficient plants, MT raised Zn concentration in root and shoot both by 16.2% and xylem sap Zn concentration by 15.6%. MT also increased root cell-wall Zn content, pectin abundance, and pectin-associated Zn by 20.0%, 39.3%, and 31.9%, respectively. At the expression level, MT moderated the induction of OsZIP4, OsZIP5, OsZIP8, OsZIP9, OsZIP10, and OsHMA3 by Zn deficiency and further repressed OsZIP1, implying feedback regulation of the Zn-homeostasis network instead of direct activation of Zn-transporter transcription. Root NO content rose by 29.8% after MT treatment, and SNP elicited increases in root and shoot Zn concentrations comparable to those associated with MT. Overall, the data suggest that an NO-associated response contributes to MT-mediated Zn-deficiency tolerance by restoring Zn homeostasis, promoting apoplastic Zn remobilization and root-to-shoot transport, and limiting oxidative damage.

The clinical management of methicillin-resistantS. aureus(MRSA) pneumonia is hindered by biofilm infections and cytokine storms. A key challenge is the lack of real-time monitoring tools for the inflammatory microenvironment during treatment. In this study, we developed a theranostic platform (CS@EAgNP) using TPP-modified chitosan to load silver sodium nitroprusside nano-aggregates (AgNP) and epigallocatechin gallate (EGCG). This system combines ultrasound (US) imaging with US-activated therapy. The CS@EAgNP nano-aggregates can act as an excellent US contrast enhancer.In vivoUS imaging indicated that AgNP generated significantly enhanced echo signals at the infection site, providing real-time and high-contrast visualization of the inflammatory foci that was unattainable with normal lung tissue. Moreover, these nano-aggregates release NO, silver and EGCG under US. The NO and silver penetrate biofilms to kill MRSA, while EGCG scavenges reactive oxygen species and modulates inflammation. This reduces pro-inflammatory cytokines like IL-6 and TNF-α, mitigating the cytokine storm.In vivoresults further confirm that this image-guided strategy achieves significant bacterial clearance and promotes lung tissue regeneration without detectable toxicity to major organs. This integrated approach bridges diagnosis and treatment, offering a promising solution for managing MRSA pneumonia.

Nitric oxide (NO) is a key signaling molecule regulating plant stress responses. In our study, the NO donor sodium nitroprusside (SNP) significantly promoted ginger seedling growth under salt stress, while treatment with the NO scavenger cPTIO resulted in an opposite response, supporting the involvement of NO in salt-stress responses. Transcriptome profiling of ginger leaves identified 2252 differentially expressed genes (DEGs), which were enriched in 55 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Further, analysis showed that 19 DEGs were enriched in plant hormone signal transduction, while 16 DEGs were enriched in starch and sugar metabolism. NO treatment increased the levels of indole-3-acetic acid (IAA), gibberellin 3 (GA3), and trans-zeatin (ZR), while reducing the levels of abscisic acid (ABA), methyl jasmonate (MeJA), and brassinosteroid (BR). Moreover, NO promoted the accumulation of soluble sugars and sucrose and enhanced the activities of key enzymes involved in starch and sucrose metabolism, thereby effectively improving osmotic adjustment. Collectively, NO improves antioxidant capacity, maintains endogenous phytohormone balance, and enhances osmotic adjustment, ultimately alleviating salt stress damage in ginger seedlings.

Aquaporins and nitric oxide (NO) are essential regulators of lateral root (LR) development in plants. However, whether an aquaporin could modulate NO-affected LR formation remains elusive. Here, we show that the tonoplast intrinsic aquaporin protein AtTIP5;1 negatively regulates NO-promoted LR development. AtTIP5;1 is highly expressed in root pericycle cells. AtTIP5;1 overexpression causes significant suppression of LR formation induced by sodium nitroprusside (SNP), an NO donor. Moreover, AtTIP5;1 overexpression results in a clear decrease in NO accumulation in the roots. The application of exogenous auxin influx inhibitor naphthoxyacetic acid boosts the effect of AtTIP5;1 on LR growth evoked by SNP, whereas the auxin efflux inhibitors N-1-naphthylphthalamic acid and 2,3,5-triiodobenzoic acid notably attenuate this effect. In addition, AtTIP5;1 overexpression leads to elevated hydrogen peroxide levels, decreased superoxide anion accumulation, enhanced superoxide dismutase activity, and reduced activities of catalase, ascorbate peroxidase, and peroxidase in roots under SNP treatment. These results suggest that AtTIP5;1 may inhibit NO-facilitated LR development by reducing NO accumulation, affecting auxin transport, and altering ROS homeostasis in Arabidopsis.