Nitric Oxide Generation Research Articles

Nitric Oxide and Proline Modulate Redox Homeostasis and Photosynthetic Metabolism in Wheat Plants under High Temperature Stress Acclimation

The effects of exogenously-sourced NO (nitric oxide, as 100 µM SNP) and proline (50 mM) in the protection of the photosynthetic performance of wheat (Triticum aestivum L.) plants against heat stress were investigated. The study focused on the mechanisms of proline accumulation, activity, gene expression of antioxidant enzymes, and NO generation. Plants were exposed to a temperature of 40 °C for 6 h per day over 15 days, then allowed to recover at 28 °C. Heat-stressed plants showed increased oxidative stress, with higher levels of H2O2 and TBARS (thiobarbituric acid reactive substances) and increased proline accumulation, ACS activity, ethylene evolution, and NO generation, which in turn leads to increased accumulation of antioxidant enzymes and reduced photosynthetic attributes. In the tested wheat cultivar, the exogenous application of SNP and proline under heat stress improved the photosynthesis and reduced oxidative stress by enhancing the enzymatic antioxidant defense system. Potentially, the promoter AOX (alternative oxidase) played a role in maintaining redox homeostasis by lowering H2O2 and TBARS levels. The genes for GR antioxidant and photosystem II core protein encoding psbA and psbB were highly up-regulated in nitric oxide and proline treated heat-stressed plants, indicating that ethylene positively impacted photosynthesis under high temperature stress. Moreover, nitric oxide supplementation under high temperature stress optimized ethylene levels to regulate the assimilation and metabolism of proline and the antioxidant system, lowering the adverse effects. The study showed that nitric oxide and proline increased high temperature stress tolerance in wheat by increasing the osmolytes accumulation and the antioxidant system, resulting in enhanced photosynthesis.

Comparison of In Vitro Estrogenic Activity of Polygoni multiflori Radix and Cynanchi wilfordii Radix via the Enhancement of ERα/β Expression in MCF7 Cells

Postmenopausal women experience several symptoms, including inflammation and a sharp rise in oxidative stress caused by estrogen deprivation. Although estrogen replacement therapy (ERT) is generally regarded as an effective treatment for menopause, it has been used less frequently due to some adverse effects and high costs. Therefore, there is an immediate need to develop an effective herbal-based treatment that is affordable for low-income populations. Acordingly, this study explored the estrogen-like properties of methanol extracts from Cynanchum wilfordii (CW) and Poligonum multiflorum (PM), two important medicinal plants in Republic of Korea, Japan, and China. Due to the similar names and morphologies of these two radixes, they are frequently confused in the marketplace. Our previous colleagues discriminated between these two plants. In this study, we investigated the estrogenic activity of PM and CW using several in vitro assays with their possible mechanism of action. First, their phytochemical contents, such as gallic acid, 2,3,5,4'-tetrahydroxystilbene-2-O-glucoside (TSG) and emodin, were quantified using high-performance liquid chromatography (HPLC). Secondly, estrogen-like activity was assessed utilizing the well-known E-screen test and gene expression analysis in estrogen receptor (ER)-positive MCF7 cells. ROS inhibition and anti-inflammatory effects were analyzed using HaCaT and Raw 264.7 cells, respectively. Our findings demonstrate that PM extracts significantly increased the expression of the estrogen-dependent genes (ERα, ERβ, pS2) and boosted MCF7 cell proliferation in comparison to CW extracts. Additionally, PM extract demonstrated a significant reduction in reactive oxygen species (ROS) production as well as an enhanced antioxidant profile compared to the CW extract. Further, the PM extract treatment significantly reduced the generation of nitric oxide (NO) in RAW 264.7 cells, a murine macrophage cell line, demonstrating the anti-inflammatory properties of the extract. Finally, this research offers an experimental foundation for the use of PM as a phytoestrogen to minimize menopausal symptoms.

Nitrogen and copper-doped saffron-based carbon dots: Synthesis, characterization, and cytotoxic effects on human colorectal cancer cells

AimDoped carbon dots (CDs) have attracted tremendous attention in cancer therapy. We aimed to synthesize copper, nitrogen-doped carbon dots (Cu, N-CDs) from saffron and investigated their effects on HCT-116 and HT-29 colorectal cancer (CRC) cells. Main methodsCDs were synthesized by hydrothermal method and characterized by transmission electron microscopy (TEM), energy-dispersive X-ray (EDX), Fourier transform infrared (FT-IR) spectroscopy, ultraviolet-visible (UV–Vis) absorption spectroscopy, and fluorescence spectroscopy. HCT-116 and HT-29 cells were incubated with saffron, N-CDs, and Cu, N-CDs for 24 and 48 h for cell viability. Cellular uptake and intracellular reactive oxygen species (ROS) were evaluated by immunofluorescence microscopy. Oil Red O staining was used to monitor lipid accumulation. Apoptosis was evaluated using acridine orange/propidium iodide (AO/PI) staining and quantitative real-time polymerase chain reaction (Q-PCR) assay. The expression of miRNA-182 and miRNA-21 was measured by Q-PCR, while the generation of nitric oxide (NO) and lysyl oxidase (LOX) activity was calculated by colorimetric methods. Key findingsCDs were successfully prepared and characterized. Cell viability decreased in the treated cells dose- and time-dependently. HCT-116 and HT-29 cells uptook Cu, N-CDs with a high level of ROS generation. The Oil Red O staining showed lipid accumulation. Concomitant with an up-regulation of apoptotic genes (p < 0.05), AO/PI staining showed increased apoptosis in the treated cells. In comparison to control cells, NO generation, and miRNA-182 and miRNA-21 expression significantly changed in the Cu, N-CDs treated cells (p < 0.05). SignificanceThe results indicated that Cu, N-CDs could inhibit CRC cells through the induction of ROS generation and apoptosis.

Recent Advances in Functional Nanomaterials for Catalytic Generation of Nitric Oxide: A Mini Review.

As a gaseous second messenger, nitric oxide (NO) plays an important role in a series of signal pathways. Research on the NO regulation for various disease treatments has aroused wide concern. However, the lack of accurate, controllable, and persistent release of NO has significantly limited the application of NO therapy. Profiting from the booming development of advanced nanotechnology, a mass of nanomaterials with the properties of controllable release have been developed to seek new and effective NO nano-delivery approaches. Nano-delivery systems that generate NO through catalytic reactions exhibit unique superiority in terms of precise and persistent release of NO. Although certain achievements have been made in the catalytically active NO delivery nanomaterials, some basic but critical issues, such as the concept of design, are of low attention. Herein, an overview of the generation of NO through catalytic reactions and the design principles of related nanomaterials are summarized. Then, the nanomaterials that generate NO through catalytic reactions are classified. Finally, the bottlenecks and perspectives are also discussed in depth for the future development of catalytical NO generation nanomaterials.

In vivo imaging of nitric oxide in the male rat brain exposed to a shock wave.

While numerous studies have suggested the involvement of cerebrovascular dysfunction in the pathobiology of blast-induced traumatic brain injury (bTBI), its exact mechanisms and how they affect the outcome of bTBI are not fully understood. Our previous study showed the occurrence of cortical spreading depolarization (CSD) and subsequent long-lasting oligemia/hypoxemia in the rat brain exposed to a laser-induced shock wave (LISW). We hypothesized that this hemodynamic abnormality is associated with shock wave-induced generation of nitric oxide (NO). In this study, to verify this hypothesis, we used an NO-sensitive fluorescence probe, diaminofluorescein-2 diacetate (DAF-2 DA), for real-time in vivo imaging of male Sprague-Dawley rats' brain exposed to a mild-impulse LISW. We observed the most intense fluorescence, indicative of NO production, along the pial arteriolar walls during the period of 10-30 min post-exposure, parallel with CSD occurrence. This post-exposure period also coincided with the early phase of hemodynamic abnormalities. While the changes in arteriolar wall fluorescence measured in rats receiving pharmacological NO synthase inhibition by nitro-L-arginine methyl ester (L-NAME) 24 h before exposure showed a temporal profile similar to that of changes observed in LISW-exposed rats with CSD, their intensity level was considerably lower; this suggests partial involvement of NOS in shock wave-induced NO production. To the best of our knowledge, this is the first real-time in vivo imaging of NO in rat brain, confirming the involvement of NO in shock-wave-induced hemodynamic impairments. Finally, we have outlined the limitations of this study and our future research directions.

Pivotal role for S-nitrosylation of DNA methyltransferase 3B in epigenetic regulation of tumorigenesis

DNA methyltransferases (DNMTs) catalyze methylation at the C5 position of cytosine with S-adenosyl-l-methionine. Methylation regulates gene expression, serving a variety of physiological and pathophysiological roles. The chemical mechanisms regulating DNMT enzymatic activity, however, are not fully elucidated. Here, we show that protein S-nitrosylation of a cysteine residue in DNMT3B attenuates DNMT3B enzymatic activity and consequent aberrant upregulation of gene expression. These genes include Cyclin D2 (Ccnd2), which is required for neoplastic cell proliferation in some tumor types. In cell-based and in vivo cancer models, only DNMT3B enzymatic activity, and not DNMT1 or DNMT3A, affects Ccnd2 expression. Using structure-based virtual screening, we discovered chemical compounds that specifically inhibit S-nitrosylation without directly affecting DNMT3B enzymatic activity. The lead compound, designated DBIC, inhibits S-nitrosylation of DNMT3B at low concentrations (IC50 ≤ 100 nM). Treatment with DBIC prevents nitric oxide (NO)-induced conversion of human colonic adenoma to adenocarcinoma in vitro. Additionally, in vivo treatment with DBIC strongly attenuates tumor development in a mouse model of carcinogenesis triggered by inflammation-induced generation of NO. Our results demonstrate that de novo DNA methylation mediated by DNMT3B is regulated by NO, and DBIC protects against tumor formation by preventing aberrant S-nitrosylation of DNMT3B.

Nitric Oxide Generation and Endothelial Progenitor Cells Recruitment for Improving Hemocompatibility and Accelerating Endothelialization of Tissue Engineering Heart Valve (Adv. Funct. Mater. 9/2023)

Nitric Oxide Generation and Endothelial Progenitor Cells Recruitment for Improving Hemocompatibility and Accelerating Endothelialization of Tissue Engineering Heart Valve (Adv. Funct. Mater. 9/2023)

Melatonin facilitates the deposition of suberin polyphenolic and lignin at wounds of potato tubers by mediating nitric oxide and reactive oxygen species

Melatonin (MT), a phytohormone, can enhance plant resistance against biotic and abiotic stresses. However, no studies have been conducted on whether MT influences suberin polyphenolic (SPP) and lignin deposition by mediating nitric oxide (NO) and reactive oxygen species (ROS). Herein, wounded tubers were treated with 50 μmol/L MT, and 25 μmol/L Nω-nitro-L-arginine (L-NNA), a specific inhibitor of nitric oxide synthase. We found that MT upregulated StNR (nitrate reductase) and StNOS (nitric oxide synthase) expression levels, and increased endogenous NO content. Additionally, MT upregulated StNOX (NADPH oxidase) expression and facilitated O2.- and H2O2 production. Moreover, MT upregulated relative expression and enhanced the activities of the main enzyme and products of the phenylpropanoid pathway, and increased peroxidase (POD) activity. MT also elevated the accumulation amount of suberin polyphenolic (SPP) and lignin at the wound tissues, resulting in less weight loss and disease index in wounded potatoes (cv. Atlantic) during healing. In contrast, L-NNA treatment suppressed the influence of MT on the generation of endogenous NO and ROS, the activity of phenylpropanoid metabolism and POD, and SPP and lignin accumulation at wounds. Taken together, MT could promote SPP and lignin deposition at wounds of potato tubers by inducing NO and ROS production and activating phenylpropanoid metabolism. However, L-NNA could eliminate the positive effect of MT.

SIRT6 regulates endothelium-dependent relaxation by modulating nitric oxide synthase 3 (NOS3)

Background and objectiveSIRT6, an NAD+-dependent protein deacetylase, is a key modulator of various biological functions. However, the precise role of SIRT6 in the regulation of endothelial function is still not fully understood. The current study sought to determine whether SIRT6 modulates NOS3 activity to regulate endothelium-dependent relaxations in the arterial wall and, if so, to investigate the potential underlying mechanism (s). MethodsApoE-/- mice and Sprague-Dawley rats had their aortic rings isolated for a vascular reactivity assay. Endothelial cells were cultured before qRT-PCR, western blot, immunoprecipitation, NO bioavailability, and acetylation/deacetylation assays were performed. ResultsSIRT6 expression was significantly reduced in the aorta of ApoE-/- mice fed a high-cholesterol diet, as was endothelium-dependent relaxation. Endothelial dysfunction could be corrected by delivering a SIRT6 overexpression construct via an adenovirus. In cultured endothelial cells, siRNA knockdown of SIRT6 decreased NOS3 catalytic activity, whereas adenoviral overexpression of SIRT6 increased NOS3-derived nitric oxide (NO) generation. SIRT6 interacted with and deacetylated human NOS3 at lysines 494, 497, and 504 of the calmodulin-binding domain, allowing calmodulin to bind to NOS3 and stimulate NOS3 activity. SIRT6 knockdown also reduced NOS3 expression by inhibiting Kruppel-Like Factor 2 (KLF2). ConclusionsWe identified SIRT6 as a new regulator of the activity of NOS3, with functional implications for endothelial-dependent relaxation.

Transcription factor AP-2 gamma/Krüppel-like factor 10 axis is involved in miR-3656-related dysfunction of endothelial cells in hypertension.

Dysfunction of endothelial cells links to microvascular rarefaction, reflecting the pathogenesis of hypertension. Our previous studies found that miR-3656 reduces nitric oxide generation and von Willebrand factor (vWF) cleavage, thereby retarding blood flow and potentially increasing blood pressure. In this paper, we investigated mechanism of transcription regulation contributing to miR-3656-damaged endothelial cells in hypertension. The effects of miR-3656 on function of endothelial cells were analyzed on the basis of proliferation, migration, tube formation, and apoptosis. The mRNA level and protein level of genes were examined using quantitative real-time polymerase chain reaction (qRT-PCR) and western blot. Dual-luciferase reporter assay was performed to confirm the binding between miR-3656 and 3' untranslated region (UTR) of transcription factor AP-2 gamma ( TFAP2C ). The binding between TFAP2C and the promoter region of Krüppel-like factor 10 ( KLF10 ) was confirmed by chromatin immunoprecipitation-qPCR assay. miR-3656 impaired the cell proliferation, migration, tube formation, and apoptosis of endothelial cells. miR-3656 inhibited the expression of TFAP2C by directly targeting 3'UTR of TFAP2C ; moreover, miR-3656-induced injury of endothelial cells was rescued by TFAP2C overexpression. Furthermore, downregulated TFAP2C decreased KLF10 expression by binding to KLF10 promoter region, and upregulated KLF10 reversed the effects of silencing TFAP2C on endothelial cells. These inhibitory processes led to interference of miR-3656 to KLF10-promoted function of endothelial cells. TFAP2C/KLF10 axis is involved in miR-3656-related dysfunction of endothelial cells in hypertension. The 3'UTR of TFAP2C and KLF10 promoter region are the hubs of the TFAP2C/KLF10 axis.

Nitric oxide synthase (NOS)-dependent nitric oxide generation is involved in putrescine-induced chilling tolerance of Anthurium andraeanum

Previous studies have demonstrated the positive effects of exogenous putrescine (Put) on the chilling tolerance of Anthurium andraeanum. In the present study, anthurium seedlings were pretreated with Put and Put in combination with NO scavenger cPTIO, nitric oxide synthase (NOS) inhibitor L-NAME, or nitrate reductase (NR) inhibitor Na2WO4 solutions for four weeks and then subjected to chilling stress (6 ºC) for three days, followed by recovery (25 ºC) for one day. NO fluorescence, endogenous NO content, and activities of NOS, NR, and copper amine oxidase (CuAO), as well as physiological indexes, were assessed during the experiment. The results were further confirmed at the transcriptional level via transcriptome sequencing and qRT-PCR. Results indicated that the application of Put prompted the accumulation of endogenous NO, and the NOS-dependent pathway probably played a vital role in the endogenous NO generation. The enhancement of chilling tolerance induced by exogenous Put could be notably reversed by the combined application of L-NAME. Moreover, transcriptome sequencing and gene expression analyses confirmed the contribution of the NOS-dependent pathway to the accumulation of endogenous NO and the effects of endogenous NO on the mediation of chilling tolerance. In summary, the NOS-dependent pathway was the primary source of endogenous NO in the Put-pretreated anthurium. Furthermore, endogenous NO was involved in the chilling response of anthurium, playing a synergistic role in the mediation of chilling tolerance with exogenous Put.

Dibenzoylmethane ameliorates adiposity-mediated neuroinflammatory response and inflammation-mediated neuronal cell death in mouse microglia and neuronal cells.

Dibenzoylmethane (DBM), a licorice-derived component, has numerous health benefits. The current study aimed to investigate the effect of DBM on adiposity-induced neuroinflammatory/oxidative response and microglial activation-induced neuronal cell damage. For this research, BV2 and HT22 cells were cultured using adipcyte- and microglia-conditioned media, respectively. DBM effectively suppressed lipopolysaccharide-induced productions in inducible nitric oxide synthase and cyclooxygenase2. Interleukin (IL)-6, monocyte chemoattractant protein-1, IL-1β, and tumor necrosis factor-α levels were also downregulated by DBM. In adipocyte-conditioned medium (ACM)-cultured BV2 cells, DBM effectively decreased ACM-induced generation of nitric oxide, reactive oxygen species, and inflammatory cytokines by activating nuclear factor erythroid 2-related factor 2/heme oxygenase-1 signaling and reducing nuclear factor kappa-light-chain-enhancer of activated B cells. In BV2-conditioned medium (BVM)-cultured neuron cells, DBM recovered the BVM-induced reduction of neuronal cell viability, thereby regulating B-cell lymphoma 2 (BCL2), BCL2-associated X (BAX), and cleaved caspase-3 protein expression. Taken together, DBM suppressed adiposity-induced inflammation/oxidative responses and inflammation-induced neuronal cell death.

Evaluation of saffron extract bioactivities relevant to skin resilience

IntroductionSaffron, Crocus sativus L, is a perennial spice herb. It has been extensively studied for its antioxidant, antidepressant, and anti-inflammatory properties, and has recently gained new interest for use in high-end cosmetics. The present work aims to elucidate the skin-protective properties of saffron in human dermal fibroblasts. MethodsThe skin-protective properties of saffron extract were evaluated in terms of tyrosinase and collagenase inhibition activities, antioxidant activity in mouse macrophage cells, collagen synthesis and hyaluronic acid synthesis and cell migration activity in primary dermal fibroblast normal human neonatal cells (HDFn). ResultsSaffron’s main phytoactive constituents - crocins, picrocrocin, safranal, and crocetin - were quantified by LC-MS at 91.0, 61.5, 3.6, and 1.9 mg/g, respectively. Saffron extract inhibited tyrosinase and collagenase with IC50 0.78 mg/mL and 0.1 mg/mL, respectively. Saffron extracts (100–200 μg/mg) suppressed reactive oxygen species and nitric oxide generation in lipopolysaccharide-stimulated RAW 264.7 murine macrophage cells and promoted collagen and hyaluronic acid synthesis in HDFn. Saffron extract at 25 μg/mg significantly promoted migration of HDFn cells (wound healing capacity) compared to control (no treatment). ConclusionThe findings highlighted the potential benefits of saffron extract relevant to skin resilience.

Bioactive Nanogels Mimicking the Antithrombogenic Nitric Oxide-Release Function of the Endothelium.

Nitric oxide (NO) plays a significant role in controlling the physiology and pathophysiology of the body, including the endothelial antiplatelet function and therefore, antithrombogenic property of the blood vessels. This property of NO can be exploited to prevent thrombus formation on artificial surfaces like extracorporeal membrane oxygenators, which when come into contact with blood lead to protein adsorption and thereby platelet activation causing thrombus formation. However, NO is extremely reactive and has a very short biological half-life in blood, so only endogenous generation of NO from the blood contacting material can result into a stable and kinetically controllable local delivery of NO. In this regards, highly hydrophilic bioactive nanogels are presented which can endogenously generate NO in blood plasma from endogenous NO-donors thereby maintaining a physiological NO flux.It is shown that NO releasing nanogels could initiate cGMP-dependent protein kinase signaling followed by phosphorylation of vasodilator-stimulated phosphoprotein in platelets. Thisprevents platelet activation and aggregation even in presence of highly potent platelet activators like thrombin, adenosine 5'-diphosphate, and U46619 (thromboxane A2 mimetic).

Cancer Therapy-Related Hypertension: A Scientific Statement From the American Heart Association.

Contemporary anticancer drugs have significantly improved cancer survival at the expense of cardiovascular toxicities, including heart disease, thromboembolic disease, and hypertension. One of the most common side effects of these drugs is hypertension, especially in patients treated with vascular endothelial growth factor inhibitors, as well as tyrosine kinase inhibitors and proteasome inhibitors. Adjunctive therapy, including corticosteroids, calcineurin inhibitors, and nonsteroidal anti-inflammatories, as well as anti-androgen hormone therapy for prostate cancer, may further increase blood pressure in these patients. Cancer therapy-induced hypertension is often dose limiting, increases cardiovascular mortality in cancer survivors, and is usually reversible after interruption or discontinuation of treatment. The exact molecular mechanisms underlying hypertension are unclear, but recent discoveries indicate an important role for reduced nitric oxide generation, oxidative stress, endothelin-1, prostaglandins, endothelial dysfunction, increased sympathetic outflow, and microvascular rarefaction. In addition, genetic polymorphisms in vascular endothelial growth factor receptors are implicated in vascular endothelial growth factor inhibitor-induced hypertension. Diagnosis, management, and follow-up of cancer therapy-induced hypertension follow national hypertension guidelines because evidence-based clinical trials specifically addressing patients who develop hypertension as a result of cancer therapy are currently lacking. Rigorous baseline assessment of patients before therapy is started requires particular emphasis on assessing and treating cardiovascular risk factors. Hypertension management follows guidelines for the general population, although special attention should be given to rebound hypotension after termination of cancer therapy. Management of these complex patients requires collaborative care involving oncologists, cardiologists, hypertension specialists, primary care professionals, and pharmacists to ensure the optimal therapeutic effect from cancer treatment while minimizing competing cardiovascular toxicities.

Two-dimensional molybdenum disulfide nanosheets evoke nitric oxide-dependent antibacterial effects.

Nanomaterials are currently being explored as novel antimicrobial agents. In this study, we first investigated the ability of two-dimensional (2D) molybdenum disulfide (MoS2) nanosheets to trigger neutrophil extracellular traps (NETs) using neutrophil-differentiated HL-60 cells as well as primary human peripheral blood neutrophils. We then addressed whether the MoS2 nanosheets themselves function as antibacterial agents. We found that MoS2 and Na2MoO4 both triggered NETs, as evidenced by the quantification of neutrophil elastase (NE) activity and immunofluorescence staining of extracellular NE, as well as scanning electron microscopy. The release of NETs was found to be nitric oxide (NO)-dependent. We also found that the MoS2 nanosheets but not the soluble salt prompted acellular NO production in the presence of NaNO2. The acellular generation of NO, suggestive of nanozyme properties of the MoS2 nanosheets, was demonstrated by electron paramagnetic resonance analysis. Electrochemical analysis using cyclic voltammetry confirmed the redox transition of the MoS2 nanosheets. Finally, MoS2 nanosheets inhibited the growth of Escherichia coli in the presence of sodium nitrate. Taken together, MoS2 nanosheets triggered cellular effects as well as acellular antibacterial effects, and we provided evidence for nitrite reductase-like properties of MoS2.

S-39-3: ENDOTHELIAL INFLAMMATION AND DYSFUNCTION INDUCED BY SARS-COV-2 SPIKE PROTEIN 1 INVOLVE ADAM17 AND INTERFERON ACTIVATION PATHWAYS

Objective: COVID19 is associated with vascular inflammation. IFN-alpha (IFNα) and IFN-lambda3 (IFNλ3) are potent cytokines produced in viral infections. Their effects involve interferon-stimulated genes (ISGs) and may influence expression of angiotensin-converting enzyme 2 (ACE2), the receptor for S-protein (S1P) of SARS-CoV-2. We hypothesized that S1P-induced immune/inflammatory responses in endothelial cells (EC) are mediated via IFN-activated pathways Design and methods: Human ECs were stimulated with S1P (1 μg/mL), IFNα (100ng/mL) or IFNλ3 (100IU/mL). Because ACE2, ADAM17 and TMPRSS2 are important for SARS-CoV-2 infection, we used inhibitors of ADAM17 (marimastat, 3.8 nM), ACE2 (MLN4760, 440pM), and TMPRSS2 (camostat, 50 μM). Gene and protein expression was investigated by real-time PCR and immunoblotting, respectively. Vascular function was assessed in mesenteric arteries from wild-type (WT) normotensive and hypertensive (LinA3) mice and in ISG15-deficient (ISG15KO) mice. Results: S1P increased expression of IFNα (3-fold), IFNλ3 (4-fold) and ISGs (2-fold) in EC (p &lt; 0.05). EC responses to IFNα (ISG15: 16-fold) were greater than to IFNλ3 (ISG15: 1.7-fold) (p &lt; 0.05). S1P increased gene expression of IL-6 (1.3-fold), TNFα (6.2-fold) and IL-1β (3.3-fold), effects that were amplified by IFNs. Only the ADAM17 inhibitor marimastat inhibited S1P effects. IFNα and IFNλ3 increase protein expression of ADAM17 (27%) and TMPRSS2 (38%). No changes were observed on ACE2 expression. This was associated with increased phosphorylation of Stat1 (134%), Stat2 (102%), ERK1/2 (42%). EC production of IL-6 was increased by IFNα (1,230pg/mL) and IFNλ3 (1,124pg/mL) vs control (591pg/mL). Nitric oxide generation and eNOS phosphorylation (Ser1177) were reduced by IFNα (40%) and IFNλ3 (40%). Vascular functional responses demonstrated that endothelium-dependent vasorelaxation (% Emax) in vessels from WT-mice stimulated with IFNα (67%) and IFNλ3 (71%) were reduced vs control (82%) (p &lt; 0.05). Responses were not altered in vessels from ISG15KO mice. Increased contraction was observed only in vessels from hypertensive mice treated with IFNα (9.1 ± 0.5mN vs control: 7.3 ± 0.3mN) (p &lt; 0.05). Conclusions: In ECs, S1P, IFNα and IFNλ3 increased ISG15 and IL-6 by mechanisms dependent on ADAM17. IFNs amplifies endothelial cell inflammatory responses and induced vascular dysfunction through ISG15-dependent mechanisms, with augmented effects in hypertension. Our findings demonstrate that S1P induces immune/inflammatory responses that may be important in endotheliitis associated with COVID-19. This may be especially important in the presence of cardiovascular risk factors, including hypertension.

Levels of nitric oxide in smokers and nonsmokers with chronic periodontitis before and after nonsurgical periodontal treatment.

The purpose of this study was to determine the level of nitric oxide (NO) in both serum and saliva of smokers and nonsmokers with chronic periodontitis before and after scaling and root planing, as well as to compare the same (NO levels) with the severity of periodontitis. Sixty people took part in the study and were divided into three groups. The control group (Group I) involved 20 patients who were nonsmokers with healthy periodontium. The other two groups included 20 patients each, where Group II was nonsmokers with chronic periodontitis and Group III was smokers with chronic periodontitis. In addition, NO generation was quantified indirectly in this study using the Griess reaction to determine the nitrite level in serum and saliva. The mean value of salivary and serum NO was higher in Group III than in Group II, and NO decreased considerably (P < 0.01) in both Groups II and III after treatment compared to before treatment. Serum and salivary NO levels can be used as a good predictor of the inflammatory condition of the periodontium in smokers.

Direct fluorescence labelling of NO inside plant cells.

Nitric oxide (NO) plays a key role in regulating plant growth, enhances nutrient uptake, and activates disease and stress tolerance mechanisms in most plants. NO is marked as a potential tool for improving the yield and quality of horticultural crop species. Research on NO in plant species can provide an abundance of valuable information regarding this. Hence, we have prepared a simple chemosensor (NPO) for the detection of endogenous NO in chickpea saplings. NPO selectively interacts with NO as determined through a chemodosimetric method to clearly show both the colorimetric and fluorometric changes. After the interaction with NO, the colorless NPO turns yellow as observed by the naked eye and shows bright cyan-blue fluorescence under a UV lamp. The 1 : 1 stoichiometric ratio between NPO and NO is determined from Job's plot resulting in a stable diazeniumdiolate product. The interaction mechanism is well established by absorption, fluorescence titration, NMR titration, HRMS, and DFT calculations. This method has successfully been employed in the plant's root and stem systems to label NO. Confocal microscopy images might help us to understand the endogenous NO generation and the mechanism that happens inside plant tissues.

Blue Light Therapy in the Management of Chronic Wounds: A Narrative Review of its Physiological Basis and Clinical Evidence

Introduction. Chronic wounds are a significant problem worldwide, with substantial cost to health care systems; thus, a minimally invasive and well-tolerated treatment is attractive. Blue light has shown promise in wound healing through the principle of photobiomodulation. Objective. This review examines the physiological effects of blue light on tissue and the hypothesis that appropriate application of blue light in conjunction with SOC improves wound healing compared with SOC alone. Methods. The authors searched in PubMed, Google Scholar, and the Cochrane Library to identify literature on the mechanism of action of blue light and then examined the clinical evidence. Results. Key physiological pathways of blue light include generation of ROS and nitric oxide, resulting in promotion of angiogenesis, reduced inflammation, and direct antimicrobial effects. These reactions are seen only at low doses; in fact, higher doses may be harmful to tissue. The only primary study with statistical analyses demonstrated wound area reduction of 51% (P =.007) in blue light–irradiated wounds compared with SOC alone. Conclusions. Blue light applied following a strict protocol is safe and shows promise in the management of chronic wounds. The current evidence is poor, however, and randomized trials are required to confirm its clinical utility.