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General mode of operation of the chemiluminescence method for detection of NO. Note: (A) Working model of chemiluniescence method for NO detection. Adapted from Bates, 33 Hetrick and Schoenfisch. 34 (B) Luciferase-based chemiluminescence method for detection of NO concentration in cell cultures. Adapted from Woldman et al. 35 NO: Nitric oxide; PMT: photomultiplier; NO 2 *: nitrogen dioxide in the excited state; hν: luminescence (h: Planck's constant, ν: frequency of photon); ONOO -: highly oxidative peroxynitrite; GTP: guanosine triphosphate; cGMP: cyclic guanosine monophosphate; PPi: pyrophosphate; APS: adenosine-5′ phosphosulfate; ATP: adenosine triphosphate; AMP: adenosine monophosphate.
Source publication
Initially being considered as an environmental pollutant, nitric oxide has gained the momentum of research since its discovery as endothelial derived growth factor in 1987. Extensive researches have revealed the various pathological and physiological roles of nitric oxide such as inflammation, vascular and neurological regulation functions. Hence,...
Contexts in source publication
Context 1
... principle of chemiluminescence detection is based on NO-triggered chemiluminescence reactions. Chemiluminescence can be realized by the oxidization of NO with ozone (O 3 ) into nitrogen dioxide in the excited state (NO 2 *), which emits a photon spontaneously when it decays back to its basal lower energy state ( Figure 4A). 2,3,7,[31][32][33] Another route to chemiluminescence is the use of chemiluminescence agent such as luminol to be excited by highly oxidative peroxynitrite (ONOO -) which is generated by the reaction of NO with H 2 O 2 . ...
Context 2
... The emission of photon or luminescence is measured by a photomultiplier. Photomultiplier converts this luminescence into electrical signal which is proportional to NO concentration ( Figure 4A). 35 The high specificity of NO detection is attributed to its unique properties, which includes its ability to exist as a gas and its quick reaction rate with ozone. ...
Context 3
... addition, Woldman et al. 35 developed a chemiluminescence method for detection of NO generation in cell cultures by a luciferin-luciferase system ( Figure 4B). The activation product (pyrophosphate) of guanylyl cyclase by NO reaction was converted to adenosine triphosphate by adenosine triphosphate sulfurylase which further excited the luciferin-luciferase system to generate chemiluminescence. ...
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Citations
... The NO microprobe was connected to PowerLab (AD Instruments) and the signal was recorded using LabChart 5. A standard curve of NO was constructed using known concentrations of SNAP and spiking them into the reaction chamber with a continuous flow of nitrogen. Once the standard curve was established, the procedure was repeated by spiking SNAP-MSs to confirm the encapsulation and release of NO from the microspheres [29,30]. ...
Heart disease is one of the leading causes of death in the United States and throughout the world. While there are different techniques for reducing or preventing the impact of heart disease, nitric oxide (NO) is administered as nitroglycerin for reversing angina or chest pain. Unfortunately, due to its gaseous and short-lived half-life, NO can be difficult to study or even administer. Therefore, controlled delivery of NO is desirable for therapeutic use. In the current study, the goal was to fabricate NO-releasing microspheres (MSs) using a donor molecule, S-Nitroso-N-Acetyl penicillamine, (SNAP), and encapsulating it in poly(ε-caprolactone) (PCL) using a single-emulsion technique that can provide sustained delivery of NO to cells over time without posing any toxicity risks. Optimization of the fabrication process was performed by varying the duration of homogenization (5, 10, and 20 min) and its effect on entrapment efficiency and size. The optimized SNAP-MS had an entrapment efficiency of ˃50%. Furthermore, we developed a modified method for NO detection by using NO microsensors to detect the NO release from SNAP-MSs in real time, showing sustained release behavior. The fabricated SNAP-MSs were tested for biocompatibility with HUVECs (human umbilical vein endothelial cells), which were found to be biocompatible. Lastly, we tested the effect of controlled NO delivery to human induced pluripotent stem-derived cardiomyocytes (hiPSC-CMs) via SNAP-MSs, which showed a significant improvement in the electrophysiological parameters and alleviated anoxic stress.
... This issue is particularly relevant given that alterations in NO concentrations can either exert beneficial effects on cellular homeostatic mechanisms or toxicity [22,23]. However, the high reactivity of NO has traditionally represented a major barrier to its measurement in biological samples [24,25]. The measurement of alternative metabolites, e.g., nitrite and nitrate, is also affected by a number of factors that have curtailed its routine use in research and clinical studies [26,27]. ...
Alterations of nitric oxide (NO) homeostasis have been described in mood disorders. However, the analytical challenges associated with the direct measurement of NO have prompted the search for alternative biomarkers of NO synthesis. We investigated the published evidence of the association between these alternative biomarkers and mood disorders (depressive disorder or bipolar disorder). Electronic databases were searched from inception to the June 30, 2023. In 20 studies, there was a trend towards significantly higher asymmetric dimethylarginine (ADMA) in mood disorders vs. controls (p = 0.072), and non-significant differences in arginine (p = 0.29), citrulline (p = 0.35), symmetric dimethylarginine (SDMA; p = 0.23), and ornithine (p = 0.42). In subgroup analyses, the SMD for ADMA was significant in bipolar disorder (p < 0.001) and European studies (p = 0.02), the SMDs for SDMA (p = 0.001) and citrulline (p = 0.038) in European studies, and the SMD for ornithine in bipolar disorder (p = 0.007), Asian (p = 0.001) and American studies (p = 0.005), and patients treated with antidepressants (p = 0.029). The abnormal concentrations of ADMA, SDMA, citrulline, and ornithine in subgroups of mood disorders, particularly bipolar disorder, warrant further research to unravel their pathophysiological role and identify novel treatments in this group (The protocol was registered in PROSPERO: CRD42023445962)
... Future experiments should also assess the extent to which Notch signaling participates in defining vascular permeability through NO regulation and whether this interaction interferes with vascular permeability and leakiness profile in pathological conditions such as PAD. Many methods are available to estimate NO in vivo and in vitro (Goshi et al., 2019), and previous works have estimated NO release by HUVECs to be within the ranges of nM to μM (Østergaard et al., 2007;Ugusman et al., 2014;Janaszak-Jasiecka et al., 2018). Additionally, in vivo measurements of NO report values of about 1 μM, with the values of EC50 for soluble guanylate cyclase (sGC) in the vascular smooth muscle as low as several nM (Chen et al., 2008). ...
Introduction: Several signaling pathways are activated during hypoxia to promote angiogenesis, leading to endothelial cell patterning, interaction, and downstream signaling. Understanding the mechanistic signaling differences between endothelial cells under normoxia and hypoxia and their response to different stimuli can guide therapies to modulate angiogenesis. We present a novel mechanistic model of interacting endothelial cells, including the main pathways involved in angiogenesis.
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... High level of NO in vivo induces the production of other reactive oxygen species (ROS) and reactive nitrogen species (RNS), which are closely related to tumors, arthritis, diabetes and neurodegenerative diseases [6,7]. Moreover, NO at the concentrations from pM to nM in tumor cells can accelerate cancer progression by promoting tumor cell growth [8,9]. Low level of NO at nanomolar concentrations is involved in both cancers [10], and central nervous system activity, such as the abnormality production during brain ischaemia [11]. ...
... 33 In the current study, NO levels did not differ between the four trials. A plausible explanation could be the fact that NO was assessed in exhaling air and not using the gold standard methodology which is the Griess assay, which is a colorimetric method for the quantification of nitrite, a stable breakdown product of NO. 34 It is well known that chronic exercise training is beneficial for health and quality of life parameters in patients with ESRD. 9,35 Specifically, chronic exercise may ameliorate muscle catabolism by promoting an anabolic milieu, thereby potentially improving the clinical sequelae of sarcopenia, such as muscle weakness, falls, fractures, frailty, IR, and immune dysfunction, in ESRD patients. ...
Hemodialysis (HD) patients suffer from multiple health problems, including severe insulin resistance. Both cold dialysis and intradialytic exercise training could elicit health benefits; however, it is still unknown whether the combination of those two approaches could enhance overall health. The current study aimed to evaluate the separate and combined acute effects of a single session of cold dialysis and intradialytic exercise in parameters related to insulin sensitivity and glucose disposal. Ten HD patients (57.2 ± 14.9 years) participated in the study. Each patient participated in four different scenarios during HD: a) typical dialysis with dialysate temperature at 37°C (TD), b) cold dialysis with dialysate temperature at 35°C, c) typical HD combined with a single exercise bout, d) cold dialysis combined with a single exercise bout. Glucose disposal and insulin resistance were assessed immediately after the end of the HD session. None of the examined parameters significantly differed between the four scenarios ( p > 0.05). However, slight numerical changes and moderate to high effect size ( d : 0.50–0.85) were observed between TD versus cold dialysis and TD versus TD + exercise in glucose and insulin disposal rates. A single session of cold and TD with intradialytic exercise may provide an “acute” time-efficient stimulus for consecutively improving glucose disposal and insulin sensitivity.
... 22 Significant hurdles in the discovery of biomarkers of NO and redox homeostasis, however, are represented by the highly reactive nature of these compounds and the influence of external factors in their analytical assessment. 23,24 This has prompted the search for stable metabolites that are closely associated with NO synthesis and oxidative stress. Several metabolites within the transsulfuration and the folic acid biochemical pathways have been shown to reflect alterations in NO synthesis and/or redox state. ...
Background
Metabolomic assessment of the transsulfuration and folic acid biochemical pathways could lead to the identification of promising biomarkers of nitric oxide dysregulation and oxidative stress in rheumatoid arthritis (RA).
Methods
We conducted a systematic review and meta‐analysis of transsulfuration (methionine, homocysteine, and cysteine) and folic acid (folic acid, vitamin B 6 , and vitamin B 12 ) metabolites in RA patients in remission and healthy controls. Electronic databases were searched from inception to 15 July 2023 for relevant articles. We assessed the risk of bias using the JBI checklist and the certainty of evidence using GRADE.
Results
In 28 eligible studies, compared to controls, RA patients had significantly higher concentrations of homocysteine (standardized mean difference, SMD = 0.74, 95% CI 0.54–0.93, p < 0.001; low certainty of evidence) and methionine (SMD = 1.00, 95% CI 0.57–1.44, p < 0.001; low certainty) and lower concentrations of vitamin B 6 (SMD = −6.62, 95% CI −9.65 to −3.60, p < 0.001; low certainty). By contrast, there were non‐significant between‐group differences in vitamin B 12 and folic acid. In meta‐regression and subgroup analysis, there were no associations between the effect size and several study and patient characteristics except for homocysteine (year of publication, C‐reactive protein, triglycerides, and analytical method) and folic acid (biological matrix).
Conclusions
The results of our study suggest that homocysteine, methionine, and vitamin B 6 are promising biomarkers to assess nitric oxide dysregulation and oxidative stress in RA. (PROSPERO registration number: CRD42023461081).
... Due to the low concentration of NO, ranging from 1 nM to 1 µM, and the extremely short half-life, estimated at less than 1 ms to multiple seconds in biological samples, 71 alternative strategies for the detection of reaction products of NO biochemistry have been developed, including colorimetric, chemiluminescence, fluorescence and electrochemical method, among others. 72 However, these methods all suffer from various shortcomings, including troublesome sample preparation, low specificity and sensitivity and high cost of instrumentation and time consumption. Therefore, more advanced quantification technologies need to be developed before pharmacokinetic characteristics of NO could be accurately determined in vivo. ...
Nitric oxide (NO), a gaseous free radical produced from L-arginine catalyzed by NO synthase, functions as an important signaling molecule in the human body. Its antiviral activity was confirmed in the 1990s, and has been studied more extensively since the outbreak of the SARS pandemic in 2003. In the fight against the ongoing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, some recent studies have revealed the potential of NO in the treatment of coronavirus disease 2019 (COVID-19). The progress in this field, including several noteworthy clinical trials of inhaled NO for the treatment of COVID-19 and the emergency approval of NO nasal spray by the regulatory agencies of Israel, Bahrain, Thailand and Indonesia for the treatment of COVID-19 pneumonia, offers a new perspective for addressing the raging coronavirus infection and greatly broadens the clinical application of NO therapy. This review aims to explore the underlying molecular mechanisms of NO-based therapy against SARS-CoV-2, including direct viral inhibition, immune regulation, and protection against pulmonary and cardiovascular symptoms. Furthermore, the potential therapeutic applications of inhaled NO, NO donors and drugs involved in the NO pathway are discussed. In the context of a global vaccination campaign and newly proposed strategy of “coexistence with COVID-19,” the advantages of NO therapies as symptomatic and adjuvant treatments are expected to deliver breakthroughs in the treatment of COVID-19.
... The Griess reaction and chemiluminescence involve the indirect quantification of by-products of reactions between NO and other species, while absorbance, fluorescence, and electron paramagnetic resonance (EPR) provide the direct measurement of adducts produced between NO and complexes, e.g., fluorescent dyes, and spin traps, respectively. [15][16][17] One of the simplest and most commonly used methods to indirectly measure NO is the Griess assay, which involves the spectrophotometric determination of its stable decomposition products, specifically nitrite (NO 2 À ), but also nitrate (NO 3 À ) following chemical reduction to nitrite. The assay is based on the Griess reaction, whereby nitrite under acidic conditions reacts with sulfanilamide to generate a diazonium ion which then combines with N-(1-napthyl)ethylenediamine to form an azo dye that can be detected spectrophotometrically at 540 nm. ...
Nitric oxide (NO) signaling plays many pivotal roles impacting almost every organ function in mammalian physiology, most notably in cardiovascular homeostasis, inflammation, and neurological regulation. Consequently, the ability to make real-time and continuous measurements of NO is a prerequisite research tool to understand fundamental biology in health and disease. Despite considerable success in the electrochemical sensing of NO, challenges remain to optimize rapid and highly sensitive detection, without interference from other species, in both cultured cells and in vivo. Achieving these goals depends on the choice of electrode material and the electrode surface modification, with graphene nanostructures recently reported to enhance the electrocatalytic detection of NO. Due to its single-atom thickness, high specific surface area, and highest electron mobility, graphene holds promise for electrochemical sensing of NO with unprecedented sensitivity and specificity even at sub-nanomolar concentrations. The non-covalent functionalization of graphene through supermolecular interactions, including π–π stacking and electrostatic interaction, facilitates the successful immobilization of other high electrolytic materials and heme biomolecules on graphene while maintaining the structural integrity and morphology of graphene sheets. Such nanocomposites have been optimized for the highly sensitive and specific detection of NO under physiologically relevant conditions. In this review, we examine the building blocks of these graphene-based electrochemical sensors, including the conjugation of different electrolytic materials and biomolecules on graphene, and sensing mechanisms, by reflecting on the recent developments in materials and engineering for real-time detection of NO in biological systems.
... Such hypothesis must be confirmed in future studies through additional assays, which might include treatments with NO scavengers, additional NO donors like GSNO, and/or monitoring of NO levels [90]. At this last regard, the use of quantitative fluorochromes, chemiluminescence, electrochemical or highly resolutive and quantitative methods like EPR (electron paramagnetic resonance) combined with NO spin traps would be of application [97]. ...
Virgin Argan Oil (VAO) is extracted from the fruits of the Moroccan endemic species argan [Argania spinosa (L.) Skeels]. It is known for its richness in polyunsaturated fatty acids and its unique composition in tocopherols conferring a panoply of pharmacological properties, documented in several studies. The aim of this study was to investigate the potential protective effect of VAO against oxidative and nitrosative stress induced in cells of the model species Tetrahymena pyriformis. As a comparison, well-known Virgin Olive Oil (VOO) from Olea europaea L. was used instead. Both oils were subjected to a preliminary analysis of phytochemicals and properties of interest. Oxidative stress in T. pyriformis cells was induced by hydrogen peroxide (H2O2) at 350µM, whereas sodium nitroprusside (SNP) nitrosative stress was induced using a 1mM concentration. Neither of these concentrations caused relevant changes in cell viability. The level of reactive oxygen species (ROS) was evaluated in the cell cultures by using H2-DCFDA dye. The activity and cell localization of the antioxidant enzymes catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPX) was evaluated as well as the levels of glutathione (GSH) and the malondialdehyde (MDA) generated after lipid peroxidation. Finally, the presence and localization of intracellular lipid droplets was assessed by using Nile red. The treatment of cultures with H2O2 produced significant increases in the activity of CAT, SOD and GPX and in the level of GSH, as also did the SNP treatment. MDA level was increased by both the treatments. VAO and VOO treatment were found to protect T. pyriformis from oxidative stress and increased cells defense in nitrosative stress. In another hand VAO and VOO decreased significantly MDA level increased by H2O2 treatment and failed after SNP treatment. The quantification of fluorescence signal obtained from immunolocalization of antioxidant enzymes confirmed the results obtained after the evaluation of their activities. Interestingly the level of ROS and the number of lipid droplets increased by H2O2 treatment was significantly decreased by VAO and VOO co-treatments. VAO and VOO represent strong antioxidants, playing an important role in protecting cells against oxidative stress.
... A significant limitation in the development of analytical platforms for the assessment of NO and biomarkers of oxidative stress in biological samples is represented by the highly reactive nature of these compounds, the relatively short half-life of NO, and the influence of other factors in the assessment of circulating NO metabolites such as nitrite and nitrate [39][40][41][42][43][44][45]. Therefore, an alternative approach consists of measuring stable metabolites within metabolic pathways that are closely associated with NO synthesis and oxidative stress. ...
There is an increasing interest in biomarkers of nitric oxide dysregulation and oxidative stress to guide management and identify new therapeutic targets in patients with chronic obstructive pulmonary disease (COPD). We conducted a systematic review and meta-analysis of the association between circulating metabolites within the arginine (arginine, citrulline, ornithine, asymmetric, ADMA, and symmetric, SDMA dimethylarginine), transsulfuration (methionine, homocysteine, and cysteine) and folic acid (folic acid, vitamin B6, and vitamin B12) metabolic pathways and COPD. We searched electronic databases from inception to 30 June 2023 and assessed the risk of bias and the certainty of evidence. In 21 eligible studies, compared to healthy controls, patients with stable COPD had significantly lower methionine (standardized mean difference, SMD = −0.50, 95% CI −0.95 to −0.05, p = 0.029) and folic acid (SMD = −0.37, 95% CI −0.65 to −0.09, p = 0.009), and higher homocysteine (SMD = 0.78, 95% CI 0.48 to 1.07, p < 0.001) and cysteine concentrations (SMD = 0.34, 95% CI 0.02 to 0.66, p = 0.038). Additionally, COPD was associated with significantly higher ADMA (SMD = 1.27, 95% CI 0.08 to 2.46, p = 0.037), SDMA (SMD = 3.94, 95% CI 0.79 to 7.08, p = 0.014), and ornithine concentrations (SMD = 0.67, 95% CI 0.13 to 1.22, p = 0.015). In subgroup analysis, the SMD of homocysteine was significantly associated with the biological matrix assessed and the forced expiratory volume in the first second to forced vital capacity ratio, but not with age, study location, or analytical method used. Our study suggests that the presence of significant alterations in metabolites within the arginine, transsulfuration, and folic acid pathways can be useful for assessing nitric oxide dysregulation and oxidative stress and identifying novel treatment targets in COPD. (PROSPERO registration number: CRD42023448036.)
