Nitric oxide (NO)-related pharmaceuticals: contemporary approaches to therapeutic NO modulation

The biologically important gaseous radical, nitric oxide (NO), is a versatile chemical entity that enters into regulatory, protective, and adverse interactions with biomolecules and cells, in some cases through NO-derived nitrogen oxide species. Both excess tissue NO and its insufficiency have been implicated in the genesis or evolution of several important disease states. The associated medical needs and commercial opportunities have fostered attempts to modulate tissue NO tone for symptomatic benefit or therapeutic gain. State-of-the-art strategies for NO modulation in contemporary drug discovery and development encompass sexual dysfunction, cardiovascular, and antiinflammatory indications. Increased understanding of NO's physiological chemistry and ways to target its pharmacology appear critical to the successful clinical exploitation of NO's diverse properties. Integration of research on both the basic science of NO's mechanistic biology and the applied science of drug discovery and development represents a millennium mandate to the pharmaceutical industry in the area of NO-related therapeutics.     

Nitric oxide (NO) and NO cycle in myocardium: molecular, biochemical and physiological aspects

The article continues the series of our publications on the problem of nitric oxide (NO) and its cyclic conversion in mammals. This review is held to analysis of nitric oxide role in regulation of cardiovascular system and in alocation of NO-synthases in myocardium. Molecular, biochemical and cytophysiological aspects that linked, with spatial localization of NO-synthases and mechanisms of NO content regulation in myocardium are considered. The results of author's investigations along the cyclic convertion of NO and literature data about compartmentalization of NO-synthases in myocardium are included in this paper. The contradictory and dissimilar facts about regulatory and toxic role of nitric oxide in cardiovascular system are represented.     

Nitric oxide (NO) and cartilage metabolism: NO effects are modulated by superoxide in response to IL-1

Nitric oxide (NO) is thought to mediate most effects of interleukin-1 (IL-1) on cartilage. In vitro evidence includes the decreased synthesis of extracellular matrix components, the abnormal cell renewal, the decreased production of IL-1 receptor antagonist, the induction of apoptosis and the enhanced sensitivity of chondrocytes to oxidative stress. Studies in NOS2(-/-) mice or administration of NO synthase inhibitors in animal models of joint disorders have confirmed its potent pathophysiological role in cartilage. Using L-NMMA (1 mM), as a NO synthase inhibitor, and CuDips (10 microM), as a SOD mimetic, we provide evidence that the inhibitory potency of IL-1beta on proteoglycan synthesis and its stimulating effect on COX-2 activity depend both on NO and O2-* production. Peroxynitrite formation is further demonstrated by the occurrence of 3-nitrotyrosines in chondrocytes stimulated in vitro with 2.5 ng/ml IL-1 and in femoral condyles of rats injected locally with 1 microg IL-1. Preliminary data suggest that such contribution of reactive oxygen species is not shared in common by IL-17, another NO-producing cytokine. We conclude that superoxide is a key modulator of NO-mediated effects in chondrocyte stimulated with IL-1 and that a combined therapy with NO synthase inhibitors and antioxidants may be promising for a full cartilage protection.     

一氧化氮自由基是气血通畅的驱动力

一氧化氮（NO）自由基是气血通畅的动力之一;抗氧化剂清除氧自由基,保护NO并且能够协助NO共同促进气血通畅,预防和保护心脑血管的健康;NO和天然抗氧化剂可以延缓衰老,还可以协同预防和治疗炎症引起的损伤;包含天然抗氧化剂的中草药,促进气血通畅,溶化血栓,是中草药通过一氧化氮自由基活血化瘀的机理。     

Nitric oxide (NO) synthase immunoreactivity in the starfish Marthasterias glacialis

The neuroendocrine system of the starfish Marthasterias glacialis was investigated immunocytochemically using antisera specific for rat neuronal, bovine aortic endothelial, and mouse macrophage, nitric oxide (NO) synthases. Immunoreactivity was detected only with the antibodies specific for the neural enzyme, in the ectoneural and hyponeural tissues of the radial nerve cords and in the basiepithelial plexus and endocrine cells of the digestive tract. The pyloric stomach showed more immunoreactive structures than the other digestive organs, with the rectal caeca showing the least activity. Immunoreactive endocrine cells were located in the cardiac and pyloric stomachs and in the pyloric caeca. Co-localization of the enzyme immunoreactivity, and the staining for NADPH-diaphorase, demonstrate the presence of NO synthase in echinoderms. These results provide further evidence that NO is a neuronal messenger of early phylogenetic origin which has been conserved throughout evolution.     

Nitric oxide (NO): a key player in the senescence of Medicago truncatula root nodules

The ectomycorrhizal fungus Tricholoma populinum is host-specific with Populus species. T. populinum has wind-dispersed progagules and may be capable of long-distance dispersal. In this study, we tested the hypothesis of a panmictic population between Scandinavia and North America. DNA sequences from five nuclear loci were used to assess phylogeographic structure and nucleotide divergence between continents. Tricholoma populinum was composed of Scandinavian and North American lineages with complete absence of shared haplotypes and only one shared nucleotide mutation. Divergence of these lineages was estimated at approx. 1.7-1.0 million yr ago (Ma), which occurred after the estimated divergence of host species Populus tremula and Populus balsamifera/Populus trichocarpa at 5 Ma. Phylogeographic structure was not observed within Scandinavian or North American lineages of T. populinum. Intercontinental divergence appears to have resulted from either allopatric isolation; a recent, rare long-distance dispersal founding event followed by genetic drift; or the response in an obligate mycorrhizal fungus with a narrow host range to contractions and expansion of host distribution during glacial and interglacial episodes within continents. Understanding present genetic variation in populations is important for predicting how obligate symbiotic fungi will adapt to present and future changing climatic conditions.     

NO是植物应激反应的信号分子

根据NO的性质和可能的产生途径,略述了生物胁迫(病原菌侵害)和干旱胁迫、盐胁迫、极端温度、机械损伤、臭氧和紫外辐射等各种非生物胁迫信号与NO信号分子的偶联及其信号的级联途径,概括了NO可能介导的生物过程,讨论了NO通过其下游信号过程对与细胞的生理影响以及该下游信号过程所涉及到的cGMP、cADPR的产生和NO与其它信号分子(ROS、SA、ABA等)的协同作用,表明胁迫诱导的NO爆发是激发、启动和装备植物细胞的重要信号级联环节,这个环节能使植物细胞处于应激状态,并迅速作出反应,形成一系列适应机制。     

Nitric oxide (NO) expression during annual reproductive activity in buffalo epididymis: a histochemical and immunocytochemical study

The buffalo is one of the few domestic animals that has a seasonal mating cycle, influenced by the photoperiod. It is known that the photoperiod regulates gonadal function probably via the pineal and/or hypothalamus-pituitary axis. Moreover, the hypothalamus (melatonin) and gonads influence the production of the signaling transmitter nitric oxide (NO), suggesting that the NO may have an important role in the regulation of gonadotropin-releasing hormone secretion. This further suggests the hypothesis that NO in the epididymis has an important role in the maturation of spermatozoa and their motility and posterior fertilization capacity. The aim of the present study is to investigate the seasonal variations in the morphology of the epididymis by means histochemical and immunocytochemical techniques. We used the NADPH-d, nitric oxide synthase (NOS) I and NOS III to clarify the relationship between epididymis function and NO signaling activity. The results of this work show that NO is present in the caput of epididymis during short photoperiods, i.e., periods of maximum gonadal activity (winter) and absent during long photoperiods, i.e., periods of gonadal regression according to the previously described role of NO in spermatozoa capacitation and motility in the caput epididymis.     

Utility of gold nanoparticles in luminescence determination of trovafloxacin: comparison of chemiluminescence and fluorescence detection

Two novel sensitive sequential injection chemiluminescence analysis and fluorescence methods for trovafloxacin mesylate detection have been developed. The methods were based on the enhancement effect of gold nanoparticles on luminol–ferricyanide–trovafloxacin and europium(III)–trovafloxacin complex systems. The optimum conditions for both detection methods were investigated. The chemiluminescence signal was emitted due to the enhanced effect of gold nanoparticles on the reaction of luminol–ferricyanide–trovafloxacin in an alkaline medium. The response was linear over a concentration range of 1.0 × 10^–9 to 1.0 × 10^–2 mol/L (%RSD = 1.3), (n = 9, r = 0.9991) with a detection limit of 1.7 × 10^–10 mol/L (S/N = 3). The weak fluorescence intensity signal of the oxidation complex of europium(III)–trovafloxacin was strongly enhanced by gold nanoparticles and detected at λ_ex = 330 and λ_em = 540 nm. Fluorescence detection enabled the determination of trovafloxacin mesylate over a linear range of 1.0 × 10^–8 to 1.0 × 10^–3 mol/L (%RSD = 1.2), (n = 6, r = 0.9993) with a detection limit of 3.3 × 10^–9 mol/L. The proposed methods were successfully applied to the determination of the studied drug in its bulk form and in pharmaceutical preparations. The results were treated statistically and compared with those obtained from other reported methods. Copyright © 2015 John Wiley & Sons, Ltd.     

Interplay between abiotic (drought) and biotic (virus) stresses in tomato plants

With climate warming, drought becomes a vital challenge for agriculture. Extended drought periods affect plant–pathogen interactions. We demonstrate an interplay in tomato between drought and infection with tomato yellow leaf curl virus (TYLCV). Infected plants became more tolerant to drought, showing plant readiness to water scarcity by reducing metabolic activity in leaves and increasing it in roots. Reallocation of osmolytes, such as carbohydrates and amino acids, from shoots to roots suggested a role of roots in protecting infected tomatoes against drought. To avoid an acute response possibly lethal for the host organism, TYLCV down-regulated the drought-induced activation of stress response proteins and metabolites. Simultaneously, TYLCV promoted the stabilization of osmoprotectants' patterns and water balance parameters, resulting in the development of buffering conditions in infected plants subjected to prolonged stress. Drought-dependent decline of TYLCV amounts was correlated with HSFA1-controlled activation of autophagy, mostly in the roots. The tomato response to combined drought and TYLCV infection points to a mutual interaction between the plant host and its viral pathogen.     

Plant metabolomics in biotic and abiotic stress: a critical overview

Abiotic and biotic stresses affect plant physiology and growth. The development of metabolomics, along with other -omics technologies, allowed in depth analysis of the reactive processes characterizing plant stress as the result of the alteration of metabolites and gene expressions. Here, we organize and interpret data from 151 studies to provide an overview about metabolomic shift after exposure to either abiotic or biotic stresses including drought, salinity, heat, heavy metal, cold, pathogens and insects. Data showed that amino acids, organic acids, sugars, and sugar alcohols quantities are influenced by stresses. Proline for example, increased in almost every stress condition, while other molecules increased or decreased depending specifically on plant tissue, plant species and type of applied stress. We concluded that although it is difficult to predict precisely what a stress will cause, some general metabolic trends can be described and improve our understanding of plant response to biotic and abiotic stresses.

     

Nitric oxide production in living neurons is modulated by sphingosine: a fluorescence microscopy study

An investigation was carried out into the possible effect of sphingosine (Sph) on nitric oxide (NO) production in living neurons. Differentiated granule cells were used in a dynamic videoimaging analysis of single cells labeled, simultaneously, with FURA-2 and the NO indicator 4,5-diaminofluorescein. The results demonstrate that Sph exerts a potent inhibitory effect on the Ca2+-dependent production of NO, without modifying the [Ca2+]i. The effect appears to be specific as neither ceramide nor Sph-1-phosphate had any effect on the NO and [Ca2+]i levels. The data demonstrate that Ca2+-dependent NO production is a specific Sph target in living granule cells, suggesting that this bioactive sphingoid plays a relevant role in neuronal NO signaling.     

Midden formation by octopuses: The role of biotic and abiotic factors

Many octopus species consume their prey in a shelter, where discarded prey items accumulate to form a midden. The shelters of Octopus bimaculatus rarely have middens. Some discarded prey items are present at 20% of the shelters of O. bimaculatus, but these do not accumulate to form middens. A field experiment using artificial middens demonstrated that currents and/or surge moved some bivalve shells, and hermit crabs rapidly removed snail shells from the middens. Snails are very important in the diet of O. bimaculatus but not most other octopus species; middens do not form around O. bimaculatus shelters because hermit crabs remove the discarded snail shells.     

Nitric oxide (NO) counteracts cadmium induced cytotoxic processes mediated by reactive oxygen species (ROS) in Brassica juncea: cross-talk between ROS, NO and antioxidant responses

Research on NO in plants has achieved huge attention in recent years mainly due to its function in plant growth and development under biotic and abiotic stresses. In the present study, we investigated Cd induced NO generation and its relationship to ROS and antioxidant regulation in Brassica juncea. Cd accumulated rapidly in roots and caused oxidative stress as indicated by increased level of lipid peroxidation and H₂O₂ thus, inhibiting the overall plant growth. It significantly decreased the root length, leaf water content and photosynthetic pigments. A rapid induction in intracellular NO was observed at initial exposures and low concentrations of Cd. A 2.74-fold increase in intracellular NO was recorded in roots treated with 25 μM Cd than control. NO effects on Malondialdehyde (MDA) content and on antioxidant system was investigated by using sodium nitroprusside (SNP), a NO donor and a scavenger, [2-(4-carboxy-2-phenyl)-4,4,5,5-tetramethylinidazoline-1-oxyl-3-oxide] (cPTIO). Roots pretreated with 5 mM SNP for 6 h when exposed to 25 μM Cd for 24 h reduced the level of proline, non-protein thiols, SOD, APX and CAT in comparison to only Cd treatments. However, this effect was almost blocked by 100 μM cPTIO pretreatment to roots for 1 h. This ameliorating effect of NO was specific because cPTIO completely reversed the effect in the presence of Cd. Thus, the present study report that NO strongly counteracts Cd induced ROS mediated cytotoxicity in B. juncea by controlling antioxidant metabolism as the related studies are not well reported in this species.     

Nitric oxide (NO)-dependent and NO-independent signaling pathways act in ABA-inhibition of stomatal opening

We investigated the role of nitric oxide (NO) in ABA-inhibition of stomatal opening in Vicia faba L. in different size dishes. When a large dish (9 cm diameter) was used, ABA induced NO synthesis and the NO scavenger reduced ABA-inhibition of stomatal opening. When a small dish (6 cm diameter) was used, ABA induced stomatal closure and inhibited stomatal opening. The NO scavenger was able to reduce ABA-induced stomatal closure, but unable to reverse ABA-inhibition of stomatal opening. Furthermore, NO was not synthesized in response to ABA, indicating that NO is not required for ABA-inhibition of stomatal opening in the small dish. These results indicated that an NO-dependent and an NO-independent signaling pathway participate in ABA signaling pathway. An NO-dependent pathway is the major player in ABA-induced stomatal closure. However, in ABA-inhibition of stomatal opening, an NO-dependent and an NO-independent pathway act: different signaling molecules participate in ABA-signaling cascade under different environmental condition.Key words: ABA, environmental condition, nitric oxide, stomata, Vicia faba LNitric oxide (NO) is a key signaling molecule in plants.^1,2 It functions in disease resistance and programmed cell death,^3,4 root development,^5,6 and plant responses to various abiotic stresses.^1,2,7,8 In addition, NO is required for stomatal closure in response to ABA in several species including Arabidopsis, Vicia faba, pea, tomato, barley, and wheat.^9–11 ABA-inhibition of stomatal opening is a distinct process from ABA-induced stomatal closure.^12,13 In V. faba, these two processes employ a similar signaling pathway; NO is also a second messenger molecule for ABA-inhibition of stomatal opening in a large dish.¹⁴ In this study, we examined the role of NO in ABA-inhibition of stomatal opening using different dish sizes. In a small dish, NO is not involved in ABA-inhibition of stomatal opening: the NO-independent signaling pathway is the major player in it.     

Nitric oxide (NO) as an intermediate in the cryptogein-induced hypersensitive response--a critical re-evaluation

A hypersensitive response (HR) was induced in tobacco leaves and cell suspensions by the fungal elicitor cryptogein, and NO production was followed by chemiluminescence and occasionally by diaminofluorescein (DAF)-fluorescence. Results from both methods were at least partly consistent, but kinetics was different. NO emission was not induced by cryptogein in leaves, whereas in cell suspensions some weak NO emission was observed, which was nitrate reductase (NR)-dependent, but not required for cell death. Nitric oxide synthase (NOS) inhibitors did not prevent cell death, but PR-1 expression was weakened. In conclusion, neither NR nor NOS appear obligatory for the cryptogein-induced HR. However, a role for NO was still suggested by the fact that the NO scavenger cPTIO prevented the HR. Unexpectedly, cPTI, the reaction product of cPTIO and NO, also impaired the HR but without scavenging NO. Thus, prevention of the HR by cPTIO is not necessarily indicative for a role of NO. Further, even a 100-fold NO overproduction (over wild type) by a nitrite reductase-deficient mutant did not interfere with the cryptogein-induced HR. Accordingly, the role of NO in the HR should be reconsidered.