Nitric Oxide Production Pathways Research Articles

High nitrate levels in skeletal muscle contribute to nitric oxide generation via a nitrate/nitrite reductive pathway in mice that lack the nNOS enzyme.

Nitric oxide (NO) is a vasodilator gas that plays a critical role in mitochondrial respiration and skeletal muscle function. NO is endogenously generated by NO synthases: neuronal NO synthase (nNOS), endothelial NO synthase (eNOS), or inducible NO synthase (iNOS). NO in skeletal muscle is partly generated by nNOS, and nNOS deficiency can contribute to muscular dystrophic diseases. However, we and others discovered an alternative nitrate/nitrite reductive pathway for NO generation: nitrate to nitrite to NO. We hypothesized that nitrate supplementation would increase nitrate accumulation in skeletal muscle and promote a nitrate/nitrite reductive pathway for NO production to compensate for the loss of nNOS in skeletal muscle. Wild-type (WT) and genetic nNOS knockout (nNOS-/-) mice were fed normal chow (386.9nmol/g nitrate) and subjected to three treatments: high-nitrate water (1g/L sodium nitrate for 7days), low-nitrate diet (46.8nmol/g nitrate for 7days), and low-nitrate diet followed by high-nitrate water for 7days each. High-nitrate water supplementation exhibited a greater and more significant increase in nitrate levels in skeletal muscle and blood in nNOS-/- mice than in WT mice. A low-nitrate diet decreased blood nitrate and nitrite levels in both WT and nNOS-/- mice. WT and nNOS-/- mice, treated with low-nitrate diet, followed by high-nitrate water supplementation, showed a significant increase in nitrate levels in skeletal muscle and blood, analogous to the increases observed in nNOS-/- mice supplemented with high-nitrate water. In skeletal muscle of nNOS-/- mice on high-nitrate water supplementation, on low-nitrate diet, and in low-high nitrate treatment, the loss of nNOS resulted in a corresponding increase in the expression of nitrate/nitrite reductive pathway-associated nitrate transporters [sialin and chloride channel 1 (CLC1)] and nitrate/nitrite reductase [xanthine oxidoreductase (XOR)] but did not show a compensatory increase in iNOS or eNOS protein and eNOS activation activity [p-eNOS (Ser1177)]. These findings suggest that a greater increase in nitrate levels in skeletal muscle of nNOS-/- mice on nitrate supplementation results from reductive processes to increase NO production with the loss of nNOS in skeletal muscle.

Low-dose radiation therapy for COVID-19

Background. Radiation therapy is one of the most effective treatments in patients with malignant neoplasms, it also plays an important role in the treatment of non-oncological di­seases. The coronavirus disease 2019 (COVID-19) pandemic has prompted a review of the long-forgotten idea of using low-dose radiation therapy (LDRT) in patients with lobar and interstitial pneumonia. Historically, from 1905 to the mid-1940s, LDRT was used to treat pneumonia, and evidence suggests that it might improve survival and provide rapid relief of respiratory symptoms. The purpose was to summarize current information about ­using low-dose radiation therapy in the treatment of patients with ­COVID-19. Materials and methods. A selection of publications was carried out, which covered information on the possibilities of using low-dose radiation therapy in the treatment of patients with COVID-19. At the first stage, a search was conducted for lite­rary sources using the keywords: COVID-19, low-dose radiation therapy. At the second stage, summaries of articles were studied and publications that did not meet the research criteria were excluded. At the third stage, the full texts of the selected articles were studied for compliance with the criteria for inclusion in the reference list and research relevance. Results. As of August 1, 2023, according to ClinicalTrials.gov, among 9,244 clinical trials associated with COVID-19, sixteen are currently being conduc­ted. They examine the effectiveness of LDRT in the treatment of patients with COVID-19 and already involve 1,038 patients. The largest number of studies is conducted in European countries. It has been shown that LDRT selectively attenuates oxidative stress to an inflammatory stimulus and inhibits the nitric oxide production pathway. This may contribute to its anti-inflammatory and vascular effects. At the same time, heme oxygenase 1 and other antioxidants are induced, they are mediated by Nrf2, which is the main regulator of endogenous antioxidant, anti-stress, analgesic reactions, and is also closely related to nicotinamide adenine dinucleotide phosphate and the metabolic pathway of pentose phosphate. Nrf2 may be critical for pain relief associated with edema formation and possibly plays a major role in the effects of LDRT, whereas direct functional effects of low-dose radiation on the autonomic nervous system and nociception are likely to be smaller. Conclusions. From 1905 to 1946, researchers published the results of more than 15 clinical studies on the effectiveness of radiotherapy for inflammatory lung diseases, which included more than 850 patients and demonstrated the high therapeutic effectiveness of radiation therapy in the indicated pathology. Today, there are 16 ongoing clinical trials in the world devoted to the study of LDRT effectiveness in the treatment of patients with COVID-19, in which 1,038 patients have already been involved. The risk of radiation-induced lung and breast cancer in a 25-year-old woman exposed to 1 Gy whole chest radiation may be as high as 5.9 and 5.5 %, respectively.

Nur77 mitigates endothelial dysfunction through activation of both nitric oxide production and anti-oxidant pathways

BackgroundNur77 belongs to the member of orphan nuclear receptor 4A family that plays critical roles in maintaining vascular homeostasis. This study aims to determine whether Nur77 plays a role in attenuating vascular dysfunction, and if so, to determine the molecular mechanisms involved. MethodsBoth Nur77 knockout (Nur77 KO) and Nur77 endothelial specific transgenic mice (Nur77-Tg) were employed to examine the functional significance of Nur77 in vascular endothelium in vivo. Endothelium-dependent vasodilatation to acetylcholine (Ach) and reactive oxygen species (ROS) production was determined under inflammatory and high glucose conditions. Expression of genes was determined by real-time PCR and western blot analysis. ResultsIn response to tumor necrosis factor alpha (TNF-α) treatment and diabetes, the endothelium-dependent vasodilatation to Ach was significantly impaired in aorta from Nur77 KO as compared with those from the wild-type (WT) mice. Endothelial specific overexpression of Nur77 markedly prevented both TNF-α- and high glucose-induced endothelial dysfunction. Compared with WT mice, after TNF-α and high glucose treatment, ROS production in aorta was significantly increased in Nur77 KO mice, but it was inhibited in Nur77-Tg mice, as determined by dihydroethidium (DHE) staining. Furthermore, we demonstrated that Nur77 overexpression substantially increased the expression of several key enzymes involved in nitric oxide (NO) production and ROS scavenging, including endothelial nitric oxide synthase (eNOS), guanosine triphosphate cyclohydrolase 1 (GCH-1), glutathione peroxidase-1 (GPx-1), and superoxide dismutases (SODs). Mechanistically, we found that Nur77 increased GCH1 mRNA stability by inhibiting the expression of microRNA-133a, while Nur77 upregulated SOD1 expression through directly binding to the human SOD1 promoter in vascular endothelial cells. ConclusionOur results suggest that Nur77 plays an essential role in attenuating endothelial dysfunction through activating NO production and anti-oxidant pathways in vascular endothelium. Targeted activation of Nur77 may provide a novel therapeutic approach for the treatment of cardiovascular diseases associated with endothelial dysfunction.

A new oxidative pathway of nitric oxide production from oximes in plants

Nitric oxide (NO) is an essential reactive oxygen species and a signal molecule in plants. Although several studies have proposed the occurrence of oxidative NO production, only reductive routes for NO production, such as the nitrate (NO-3) -upper-reductase pathway, have been evidenced to date in land plants. However, plants grown axenically with ammonium as the sole source of nitrogen exhibit contents of nitrite and NO3−, evidencing the existence of a metabolic pathway for oxidative production of NO. We hypothesized that oximes, such as indole-3-acetaldoxime (IAOx), a precursor to indole-3-acetic acid, are intermediate oxidation products in NO synthesis. We detected the production of NO from IAOx and other oximes catalyzed by peroxidase (POD) enzyme using both 4-amino-5-methylamino-2′,7′-difluorescein fluorescence and chemiluminescence. Flavins stimulated the reaction, while superoxide dismutase inhibited it. Interestingly, mouse NO synthase can also use IAOx to produce NO at a lower rate than POD. We provided a full mechanism for POD-dependent NO production from IAOx consistent with the experimental data and supported by density functional theory calculations. We showed that the addition of IAOx to extracts from Medicago truncatula increased the in vitro production of NO, while in vivo supplementation of IAOx and other oximes increased the number of lateral roots, as shown for NO donors, and a more than 10-fold increase in IAOx dehydratase expression. Furthermore, we found that in vivo supplementation of IAOx increased NO production in Arabidopsis thaliana wild-type plants, while prx33-34 mutant plants, defective in POD33-34, had reduced production. Our data show that the release of NO by IAOx, as well as its auxinic effect, explain the superroot phenotype. Collectively, our study reveals that plants produce NO utilizing diverse molecules such as oximes, POD, and flavins, which are widely distributed in the plant kingdom, thus introducing a long-awaited oxidative pathway to NO production in plants. This knowledge has essential implications for understanding signaling in biological systems.

An electrochemical sensing platform for biomonitoring the potential toxicity of plastic particles by detecting NO

Plastic particles are common pollutants in our daily lives and cause a range of potential toxicities when ingested by living organisms. However, the complex morphology of plastic particles in the natural environment makes it difficult to assess their potential toxicity using conventional ecological monitoring techniques. Therefore, we constructed an electrochemical sensor based on gold/multi-wall carbon nanotube-Fe(III) meso-tetra (4-carboxyphenyl) porphyrin/Nafion (Au/MWCNT-FeTCP/Nafion) composite material to evaluate the potential toxicity of plastic particles through the detection of nitric oxide (NO). The sensor exhibited good catalytic performance for NO with a linear relationship within 0.1–––0.7 μmol/L and 0.8–––5 μmol/L and a detection limit of 0.08 μM. We used this sensor to evaluate the changes in NO released by cells and tissues exposed to polystyrene (PS) plastic particles which showed that as the concentration of PS incubated with cells increased and the incubation time prolonged, the level of NO released by cells also increased. Notably, we combined immunofluorescence, immunoblotting, and electrochemical analysis methods simultaneously to reveal the cellular signaling pathway of NO production and apoptosis induced by PS. Furthermore, the effectiveness of the sensor in biomonitoring the oxidative damage of PS plastic particles on NO was further verified at the tissue level by constructing a mouse model treated with PS plastic particles. The sensor presented here offers a promising approach to study the biotoxicity of plastic particles and may contribute to the development of novel solutions for mitigating their hazardous effects on the human health.

Supplementation with Nitric Oxide Precursors for Strength Performance: A Review of the Current Literature.

Nitric-oxide-stimulating dietary supplements are widely available and marketed to strength athletes and weightlifters seeking to increase muscle performance and augment training adaptations. These supplements contain ingredients classified as nitric oxide (NO) precursors (i.e., "NO boosters"). Endogenous NO is generated via a nitric oxide synthase (NOS)-dependent pathway and a NOS-independent pathway that rely on precursors including L-arginine and nitrates, with L-citrulline serving as an effective precursor of L-arginine. Nitric oxide plays a critical role in endothelial function, promoting relaxation of vascular smooth muscle and subsequent dilation which may favorably impact blood flow and augment mechanisms contributing to skeletal muscle performance, hypertrophy, and strength adaptations. The aim of this review is to describe the NO production pathways and summarize the current literature on the effects of supplementation with NO precursors for strength and power performance. The information will allow for an informed decision when considering the use of L-arginine, L-citrulline, and nitrates to improve muscular function by increasing NO bioavailability.

A direct numerical simulation study on NO and N[formula omitted]O formation in turbulent premixed ammonia/hydrogen/nitrogen-air flames

The formation of nitric oxide (NO) and nitrous oxide (N2O) in turbulent NH3/H2/N2-air premixed flames is investigated utilizing three-dimensional Direct Numerical Simulation (DNS) with detailed chemical kinetics in order to gain insight on the role of equivalence ratio and pressure. To this end, results from five large-scale DNS in temporal jet and shear layer configurations with equivalence ratios of 0.45, 0.9 and 1.1 (at 1 bar) and pressures of 1 and 10 bar (at an equivalence ratio of 0.45) are presented. Statistics of the temporal evolution of global formation rates of NO and N2O are analyzed. Overall, an increase of NO production (compared to heat release) is observed, except for the lean high-pressure case, which sustains lower NO but increased N2O net production rates. Time-instants of peak NO and N2O are analyzed in more detail, including progress variable and curvature conditioned statistics that reveal the impact of local flame geometry on NO production pathways and key chemical reactions involving N2O. It is found that the production of NO generally displays a high dependence on the local flame geometry, while the formation of N2O is less sensitive to curvature, except at high pressure. These observations are discussed in light of differences in thermo-diffusive effects and the topologically-driven increased availability of certain key radicals, e.g., atomic hydrogen in the consumption of N2O.

Luteolin alleviates vascular dysfunctions in CLP-induced polymicrobial sepsis in mice.

Luteolin, a naturally occurring flavonoid, is thought to have health-promoting properties as a part of human diet and has been reported to possess a wide range of pharmacological activities. Therefore, the present study was undertaken to evaluate the effect of luteolin pre-treatment on vascular dysfunctions in sepsis induced by caecal ligation and puncture (CLP) in the mouse model. Mice were divided into four groups: sham, luteolin plus sham, CLP, and luteolin plus CLP. Luteolin was administered (0.2mg/kg body weight) intraperitoneally one hour (h) before CLP surgery in mice. 20 ± 2h post CLP surgery, the isolated thoracic aorta of mice was assessed for its vascular reactivity to noradrenaline (NA) and acetylcholine (ACh). To explore the underlying mechanism, aortic mRNA expressions of α1D adrenoceptors, eNOS and iNOS were investigated. In mice with CLP-induced sepsis luteolin pre-treatment markedly increased the survival time and attenuated serum lactate level. The CLP group manifested the reduced vascular reactivity to NA and this deficit was restored by luteolin pre-treatment. However, luteolin pre-treatment did not improve α1D adrenoceptors down-regulation observed in septic mice aorta. In the presence of 1400W, the NA contractile response was significantly restored in CLP mice aortic tissue in comparison with the respective control of septic mice and further enhanced in the presence of luteolin. Luteolin reduced the iNOS mRNA expression and iNOS-derived nitrite production. Pre-treatment with luteolin restored the endothelial dysfunction in septic mice aorta by improving eNOS mRNA expression and enhanced eNOS-derived nitric oxide (NO) production in septic mice aorta and aortic iNOS gene expression and inducible NO production. The present study suggests that the vasoplegic state to NA in aorta was restored through the iNOS pathway and endothelial dysfunction was reversed via eNOS and NO production pathway.

The Role of Nitric Oxide Signaling in Plant Responses to Cadmium Stress.

Nitric oxide (NO) is a widely distributed gaseous signaling molecule in plants that can be synthesized through enzymatic and non-enzymatic pathways and plays an important role in plant growth and development, signal transduction, and response to biotic and abiotic stresses. Cadmium (Cd) is a heavy metal pollutant widely found in the environment, which not only inhibits plant growth but also enters humans through the food chain and endangers human health. To reduce or avoid the adverse effects of Cd stress, plants have evolved a range of coping mechanisms. Many studies have shown that NO is also involved in the plant response to Cd stress and plays an important role in regulating the resistance of plants to Cd stress. However, until now, the mechanisms by which Cd stress regulates the level of endogenous NO accumulation in plant cells remained unclear, and the role of exogenous NO in plant responses to Cd stress is controversial. This review describes the pathways of NO production in plants, the changes in endogenous NO levels in plants under Cd stress, and the effects of exogenous NO on regulating plant resistance to Cd stress.

N2O and NO production and functional microbes responding to biochar aging process in an intensified vegetable soil

Vegetable soils with high nitrogen input are hotspots of nitrous oxide (N2O) and nitric oxide (NO), and biochar amended to soil has been documented to effectively decrease N2O and NO emissions. However, the aging effects of biochar on soil N2O and NO production and the relevant mechanisms are not thoroughly understood. A15N tracing microcosm study was conducted to clarify the responses of N2O and NO production pathways to the biochar aging process in vegetable soil. The results showed that autotrophic nitrification was the predominant source of N2O production. Biochar aging increased the O-containing functional groups while lowering the aromaticity and pore size. Fresh biochar enhanced the AOB-amoA gene abundance and obviously stimulated N2O production by 15.5% via autotrophic nitrification and denitrification. In contrast, field-aged biochar markedly weakened autotrophic nitrification and denitrification and thus decreased N2O production by 17.0%, as evidenced by the change in AOB-amoA and nosZI gene abundances. However, the amendment with artificially lab-aged biochar had no effect on N2O production. With the extension of aging time, biochar application reduced the soil NO production dominated by nitrification. Changes in the N2O and NO fluxes were closely associated with soil NH4+-N and NO2−-N contents, indicating that autotrophic nitrification played a critical role in NO production. Overall, our study demonstrated that field-aged biochar suppressed N2O production via autotrophic nitrification and denitrification by regulating associated functional genes, but not for lab-aged biochar or fresh biochar. These findings improved our insights regarding the implications of biochar aging on N2O and NO mitigation in vegetable soils.

Effects of soil moisture on microbial processes of soil nitrogen gases production under anaerobic conditions

Gaseous nitrogen (N) emission [nitric oxide (NO), nitrous oxide (N2O), and nitrogen (N2)] is an important pathway of soil N loss. Nitrification and denitrification are the main processes of gaseous N production in soil. However, the contribution of heterotrophic nitrification, co-denitrification, and anammox to gaseous N production remains uncertain. In a laboratory soil incubation experiment, we used the 15N labelling and pairing technique, combining the nitrification inhibitor dicyandiamide (DCD), to quantify the contribution of different microbial processes to soil NO, N2O and N2 production under anaerobic conditions. The results showed that after 24 h anaerobic incubation, the highest total 15N recovery of three gases occurred at 65% water filled pore space (WFPS), accounting for 20.0% of total added 15N. Denitrification contributed 49.9%-94.1%, 29.0%-84.7%, and 58.2%-85.8% to the production of NO, N2O and N2 respectively, suggesting that denitrification was the predominant process of those three N gases emission. Heterotrophic nitrification was an important pathway of NO and N2O production, particularly at conditions with low soil water content (10% WFPS), with its contribution to those two N gases production being 50.1% and 42.8%, respectively. Co-denitrification contributed 10.6%-30.7% of N2O production. For N2 production, the total contribution of co-denitrification and anammox was 14.2%-41.8%. The role of co-denitrification can not be ignored for N2O and N2 production. Our results demonstrated that the 15N labelling and pairing technique is a promising tool to quantify the contribution of different microbial processes to gaseous N loss.

Nitrogen isotopic fractionations during nitric oxide production in an agricultural soil

Abstract. Nitric oxide (NO) emissions from agricultural soils play a critical role in atmospheric chemistry and represent an important pathway for loss of reactive nitrogen (N) to the environment. With recent methodological advances, there is growing interest in the natural-abundance N isotopic composition (δ15N) of soil-emitted NO and its utility in providing mechanistic information on soil NO dynamics. However, interpretation of soil δ15N-NO measurements has been impeded by the lack of constraints on the isotopic fractionations associated with NO production and consumption in relevant microbial and chemical reactions. In this study, anoxic (0 % O2), oxic (20 % O2), and hypoxic (0.5 % O2) incubations of an agricultural soil were conducted to quantify the net N isotope effects (15η) for NO production in denitrification, nitrification, and abiotic reactions of nitrite (NO2-) using a newly developed δ15N-NO analysis method. A sodium nitrate (NO3-) containing mass-independent oxygen-17 excess (quantified by a Δ17O notation) and three ammonium (NH4+) fertilizers spanning a δ15N gradient were used in soil incubations to help illuminate the reaction complexity underlying NO yields and δ15N dynamics in a heterogeneous soil environment. We found strong evidence for the prominent role of NO2- re-oxidation under anoxic conditions in controlling the apparent 15η for NO production from NO3- in denitrification (i.e., 49 ‰ to 60 ‰). These results highlight the importance of an under-recognized mechanism for the reversible enzyme NO2- oxidoreductase to control the N isotope distribution between the denitrification products. Through a Δ17O-based modeling of co-occurring denitrification and NO2- re-oxidation, the 15η for NO2- reduction to NO and NO reduction to nitrous oxide (N2O) were constrained to be 15 ‰ to 22 ‰ and −8 ‰ to 2 ‰, respectively. Production of NO in the oxic and hypoxic incubations was contributed by both NH4+ oxidation and NO3- consumption, with both processes having a significantly higher NO yield under O2 stress. Under both oxic and hypoxic conditions, NO production from NH4+ oxidation proceeded with a large 15η (i.e., 55 ‰ to 84 ‰) possibly due to expression of multiple enzyme-level isotopic fractionations during NH4+ oxidation to NO2- that involves NO as either a metabolic byproduct or an obligatory intermediate for NO2- production. Adding NO2- to sterilized soil triggered substantial NO production, with a relatively small 15η (19 ‰). Applying the estimated 15η values to a previous δ15N measurement of in situ soil NOx emission (NOx=NO+NO2) provided promising evidence for the potential of δ15N-NO measurements in revealing NO production pathways. Based on the observational and modeling constraints obtained in this study, we suggest that simultaneous δ15N-NO and δ15N-N2O measurements can lead to unprecedented insights into the sources of and processes controlling NO and N2O emissions from agricultural soils.

Nitrate reductases and hemoglobins control nitrogen-fixing symbiosis by regulating nitric oxide accumulation.

The interaction between legumes and rhizobia leads to the establishment of a symbiotic relationship between plant and bacteria. This is characterized by the formation of a new organ, the nodule, which facilitates the fixation of atmospheric nitrogen (N2) by nitrogenase through the creation of a hypoxic environment. Nitric oxide (NO) accumulates at each stage of the symbiotic process. NO is involved in defense responses, nodule organogenesis and development, nitrogen fixation metabolism, and senescence induction. During symbiosis, either successively or simultaneously, NO regulates gene expression, modulates enzyme activities, and acts as a metabolic intermediate in energy regeneration processes via phytoglobin-NO respiration and the bacterial denitrification pathway. Due to the transition from normoxia to hypoxia during nodule formation, and the progressive presence of the bacterial partner in the growing nodules, NO production and degradation pathways change during the symbiotic process. This review analyzes the different source and degradation pathways of NO, and highlights the role of nitrate reductases and hemoproteins of both the plant and bacterial partners in the control of NO accumulation.

Enhanced Nitric Oxide Synthesis Through Nitrate Supply Improves Drought Tolerance of Sugarcane Plants.

Nitric oxide (NO) is an important signaling molecule associated with many biochemical and physiological processes in plants under stressful conditions. Nitrate reductase (NR) not only mediates the reduction of NO3− to NO2− but also reduces NO2− to NO, a relevant pathway for NO production in higher plants. Herein, we hypothesized that sugarcane plants supplied with more NO3− as a source of N would produce more NO under water deficit. Such NO would reduce oxidative damage and favor photosynthetic metabolism and growth under water limiting conditions. Sugarcane plants were grown in nutrient solution and received the same amount of nitrogen, with varying nitrate:ammonium ratios (100:0 and 70:30). Plants were then grown under well-watered or water deficit conditions. Under water deficit, plants exhibited higher root [NO3−] and [NO2−] when supplied with 100% NO3−. Accordingly, the same plants also showed higher root NR activity and root NO production. We also found higher photosynthetic rates and stomatal conductance in plants supplied with more NO3−, which was associated with increased root growth. ROS accumulation was reduced due to increases in the activity of catalase in leaves and superoxide dismutase and ascorbate peroxidase in roots of plants supplied with 100% NO3− and facing water deficit. Such positive responses to water deficit were offset when a NO scavenger was supplied to the plants, thus confirming that increases in leaf gas exchange and plant growth were induced by NO. Concluding, NO3− supply is an interesting strategy for alleviating the negative effects of water deficit on sugarcane plants, increasing drought tolerance through enhanced NO production. Our data also provide insights on how plant nutrition could improve crop tolerance against abiotic stresses, such as drought.

Effects of NO/cGMP inhibitors in a rat model of anaphylactoid shock

Anaphylactic shock can be defined as an acute syndrome, and it is the most severe clinical manifestation of allergic diseases. Anaphylactoid reactions are similar to anaphylactic events but differ in the pathophysiological mechanism. Nitric oxide (NO) inhibitors during anaphylaxis suggest that NO might decrease the signs and symptoms of anaphylaxis but exacerbate associated vasodilation. Therefore, blocking the effects of NO on vascular smooth muscle by inhibiting the guanylate cyclase (GC) would be a reasonable strategy. This study aimed to investigate the effects of NO/cGMP pathway inhibitors methylene blue (MB), Nω-nitro-L-arginine methyl ester hydrochloride (L-NAME), and indigo carmine (IC) in shock induced by compound 48/80 (C48/80) in rats. The effect was assessed by invasive blood pressure measurement. Shock was initiated by C48/80 intravenous bolus injection 5 min before (prophylactic) or after (treatment) the administration of the inhibitors MB (3 mg/kg), L-NAME (1 mg/kg), and IC (3 mg/kg). Of the groups that received drugs as prophylaxis for shock, only the IC group did not present the final systolic blood pressure (SBP) better than the C48/80 group. Regarding shock treatment with the drugs tested, all groups had the final SBP similar to the C48/80group. Altogether, our results suggested that inhibition of GC and NO synthase in NO production pathway was not sufficient to revert hypotension or significantly improve survival.

Semen Cuscutae Administration Improves Hepatic Lipid Metabolism and Adiposity in High Fat Diet-Induced Obese Mice.

Since arginase has been shown to compete with nitric oxide (NO) synthase, emerging evidence has reported that arginase inhibition improves obesity by increasing NO production. Semen cuscutae (SC), which is a well-known Chinese medicine, has multiple biological functions such as anti-oxidant function and immune regulation. In this study, we investigated whether the SC as a natural arginase inhibitor influences hepatic lipid abnormalities and whole-body adiposity in high-fat diet (HFD)-induced obese mice. The lipid accumulation was significantly reduced by SC treatment in oleic acid-induced hepatic steatosis in vitro. Additionally, SC supplementation substantially lowered HFD-induced increases in arginase activity and weights of liver and visceral fat tissue, while increasing hepatic NO. Furthermore, elevated mRNA expressions of sterol regulatory element-binding transcription factor 1 (SREBP-1c), fatty-acid synthase (FAS), peroxisome proliferator-activated receptor-gamma (PPAR-γ)1, and PPAR-γ2 in HFD-fed mice were significantly attenuated by SC supplementation. Taken together, SC, as a novel natural arginase inhibitor, showed anti-obesity properties by modulating hepatic arginase and NO production and metabolic pathways related to hepatic triglyceride (TG) metabolism.

Nitric oxide deficiency decreases C-repeat binding factor-dependent and -independent induction of cold acclimation.

Plant tolerance to freezing temperatures is governed by endogenous components and environmental factors. Exposure to low non-freezing temperatures is a key factor in the induction of freezing tolerance in the process called cold acclimation. The role of nitric oxide (NO) in cold acclimation was explored in Arabidopsis using triple nia1nia2noa1-2 mutants that are impaired in the nitrate-dependent and nitrate-independent pathways of NO production, and are thus NO deficient. Here, we demonstrate that cold-induced NO accumulation is required to promote the full cold acclimation response through C-repeat Binding Factor (CBF)-dependent gene expression, as well as the CBF-independent expression of other cold-responsive genes such as Oxidation-Related Zinc Finger 2 (ZF/OZF2). NO deficiency also altered abscisic acid perception and signaling and the cold-induced production of anthocyanins, which are additional factors involved in cold acclimation.

Apelin intensifies vascular relaxation through activation of the endothelial nitric oxide production pathway in rats with metabolic syndrome

Apelin intensifies vascular relaxation through activation of the endothelial nitric oxide production pathway in rats with metabolic syndrome

Erythrocytic bioactivation of nitrite and its potentiation by far-red light

BackgroundNitrite is reduced by heme-proteins and molybdenum-containing enzymes to form the important signaling molecule nitric oxide (NO), mediating NO signaling. Substantial evidence suggests that deoxygenated hemoglobin within red blood cells (RBCs) is the main erythrocytic protein responsible for mediating nitrite-dependent NO signaling. In other work, infrared and far red light have been shown to have therapeutic potential that some attribute to production of NO. Here we explore whether a combination of nitrite and far red light treatment has an additive effect in NO-dependent processes, and whether this effect is mediated by RBCs. Methods and resultsUsing photoacoustic imaging in a rat model as a function of varying inspired oxygen, we found that far red light (660 nm, five min. exposure) and nitrite feeding (three weeks in drinking water at 100 mg/L) each separately increased tissue oxygenation and vessel diameter, and the combined treatment was additive. We also employed inhibition of human platelet activation measured by flow cytometry to assess RBC-dependent nitrite bioactivation and found that far red light dramatically potentiates platelet inhibition by nitrite. Blocking RBC-surface thiols abrogated these effects of nitrite and far-red light. RBC-dependent production of NO was also shown to be enhanced by far red light using a chemiluminescence-based nitric oxide analyzer. In addition, RBC-dependent bioactivation of nitrite led to prolonged lag times for clotting in platelet poor plasma that was enhanced by exposure to far red light. ConclusionsOur results suggest that nitrite leads to the formation of a photolabile RBC surface thiol-bound species such as an S-nitrosothiol or heme-nitrosyl (NO-bound heme) for which far red light enhances NO signaling. These findings expand our understanding of RBC-mediated NO production from nitrite. This pathway of NO production may have therapeutic potential in several applications including thrombosis, and, thus, warrants further study.

Redefining nitric oxide production in legume nodules through complementary insights from electron paramagnetic resonance spectroscopy and specific fluorescent probes.

Nitric oxide (NO) is a signaling molecule with multiple functions in plants. Given its critical importance and reactivity as a gaseous free radical, we have examined NO production in legume nodules using electron paramagnetic resonance (EPR) spectroscopy and the specific fluorescent dye 4,5-diaminofluorescein diacetate. Also, in this context, we critically assess previous and current views of NO production and detection in nodules. EPR of intact nodules revealed that nitrosyl-leghemoglobin (Lb2+NO) was absent from bean or soybean nodules regardless of nitrate supply, but accumulated in soybean nodules treated with nitrate that were defective in nitrite or nitric oxide reductases or that were exposed to ambient temperature. Consequently, bacteroids are a major source of NO, denitrification enzymes are required for NO homeostasis, and Lb2+NO is not responsible for the inhibition of nitrogen fixation by nitrate. Further, we noted that Lb2+NO is artifactually generated in nodule extracts or in intact nodules not analyzed immediately after detachment. The fluorescent probe detected NO formation in bean and soybean nodule infected cells and in soybean nodule parenchyma. The NO signal was slightly decreased by inhibitors of nitrate reductase but not by those of nitric oxide synthase, which could indicate a minor contribution of plant nitrate reductase and supports the existence of nitrate- and arginine-independent pathways for NO production. Together, our data indicate that EPR and fluorometric methods are complementary to draw reliable conclusions about NO production in plants.