Nanoencapsulation improves the protective effects of a nitric oxide donor on drought-stressed Heliocarpus popayanensis seedlings.

Despite the important role played by nitric oxide (NO) in plants subjected to abiotic stress, NO donors application to induce drought tolerance in neotropical tree seedlings has not yet been tested. It is also worth investigating whether NO bioactivity in drought-stressed seedlings could be potentiated by NO donors nanoencapsulation. The aim of the current study is to evaluate the effects of chitosan nanoparticles (NPs) containing S-nitroso-mercaptosuccinic acid (S-nitroso-MSA) on drought-stressed seedlings of neotropical tree species Heliocarpus popayanensis Kunth in comparison to free NO donor and NPs loaded with non-nitrosated MSA. Nanoencapsulation slowed down NO release from S-nitroso-MSA, and nanoencapsulated S-nitroso-MSA yielded 2- and 1.6-fold higher S-nitrosothiol levels in H. popayanensis roots and leaves, respectively, than the free NO donor. S-nitroso-MSA has prevented drought-induced CO2 assimilation inhibition, regardless of nanoencapsulation, but the nanoencapsulated NO donor has induced earlier ameliorative effect. Both NO and MSA have decreased oxidative stress in H. popayanensis roots, but this effect was not associated with antioxidant enzyme induction, with higher seedling biomass, or with proline and glycine betaine accumulation. Nanoencapsulated S-nitroso-MSA was the only formulation capable of increasing leaf relative water content in drought-stressed plants (from 32.3% to 60.5%). In addition, it induced root hair formation (increase by 36.6% in comparison to well-hydrated plants). Overall, results have evidenced that nanoencapsulation was capable of improving the protective effect of S-nitroso-MSA on H. popayanensis seedlings subjected to drought stress, a fact that highlighted the potential application of NO-releasing NPs to obtain drought-tolerant tree seedlings for reforestation programs.

Comparison of foliar spray and soil irrigation of biogenic CuO nanoparticles (NPs) on elemental uptake and accumulation in lettuce.

Nanoparticles (NPs) can be used in several ways in agriculture, including increasing production rates and improving nutritional values in plants. The present study aims to clarify how biogenic copper oxide nanoparticles (CuO NPs) applied by two routes of exposure (foliar spray and soil irrigation) affect the elemental uptake by lettuce. In vivo experiments using lettuce (n = 4) were performed with CuO NPs in comparison with copper salt (CuSO4), considering a final mass added of 20 mg of CuO per plant. The elemental composition of roots was mostly affected by the soil irrigation exposure for both Cu forms (NPs and salt). Neither Cu form added by soil irrigation was translocated to leaves. Copper concentration in leaves was mainly affected by foliar spray exposure for both Cu forms (NPs and salt). All Cu forms through foliar spray were sequestered in the leaves and no translocation to roots was observed. Foliar spray of CuO NPs caused no visual damage in leaves, resulted in less disturbance of elemental composition, and improved dry weight, number of leaves, CO2 assimilation, and the levels of K, Na, S, Ag, Cd, Cr, Cu, and Zn in leaves without causing significant changes in daily intake of most elements, except for Cu. Although Cu concentration increased in leaves by foliar spray of CuO NPs, it remained safe for consumption.

Characterization of the Oxidative Stress in Renal Ischemia/Reperfusion-Induced Cardiorenal Syndrome Type 3.

In kidney disease (KD), several factors released into the bloodstream can induce a series of changes in the heart, leading to a wide variety of clinical situations called cardiorenal syndrome (CRS). Reactive oxygen species (ROS) play an important role in the signaling and progression of systemic inflammatory conditions, as observed in KD. The aim of the present study was to characterize the redox balance in renal ischemia/reperfusion-induced cardiac remodeling. C57BL/6 male mice were subjected to occlusion of the left renal pedicle, unilateral, for 60 min, followed by reperfusion for 8 and 15 days, respectively. The following redox balance components were evaluated: catalase (CAT), superoxide dismutase (SOD), total antioxidant capacity (FRAP), NADPH oxidase (NOX), nitric oxide synthase (NOS), hydrogen peroxide (H2O2), and the tissue bioavailability of nitric oxide (NO) such as S-nitrosothiol (RSNO) and nitrite (NO2 -). The results indicated a process of renoprotection in both kidneys, indicated by the reduction of cellular damage and some oxidant agents. We also observed an increase in the activity of antioxidant enzymes, such as SOD, and an increase in NO bioavailability. In the heart, we noticed an increase in the activity of NOX and NOS, together with increased cell damage on day 8, followed by a reduction in protein damage on day 15. The present study concludes that the kidneys and heart undergo distinct processes of damage and repair at the analyzed times, since the heart is a secondary target of ischemic kidney injury. These results are important for a better understanding of the cellular mechanisms involved in CRS.

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 copper oxide nanoparticles on growth of lettuce (Lactuca sativa L.) seedlings and possible implications of nitric oxide in their antioxidative defense.

Copper oxide nanoparticles (CuO NPs) have been extensively explored for use in agriculture. Previous studies have indicated that application of CuO NPs might be promising for development and conservation of plants, pest control, and for the recovery of degraded soils. However, depending on the applied concentration copper can cause phytotoxic effects. In this work, biosynthesized CuO NPs (using green tea extract) were evaluated on their effects on lettuce (Lactuca sativa L.) seedling growth, which were exposed at concentrations ranged between 0.2 and 300 µg mL-1. From the biosynthesized were obtained ultra-small CuO NPs (~ 6.6 nm), with high stability in aqueous suspension. Toxicity bioassays have shown that at low concentrations (up to 40 µg mL-1), CuO NPs did not affect or even enhanced the seed germination. At higher concentrations (higher than 40 µg mL-1), inhibition of seed germination and radicle growth ranging from 35 to 75% was observed. With the increase of CuO NPs concentrations, nitrite and S-nitrosothiols levels in radicles increased, whereas superoxide dismutase and total antioxidant activities decreased. The nitrite and S-nitrosothiols levels in lettuce radicles showed a direct dose response to CuO NP application, which may indicate nitric oxide-dependent signaling pathways in the plant responses. Therefore, the results demonstrated that at low concentrations (≤ 20 µg mL-1) of CuO NPs, beneficial effects are obtained from seedlings, enhancing plant growth, and the involvement of nitric oxide signaling in the phytotoxic effects induced by high concentration of this formulation. Graphical abstract.

Effects of nitric oxide-releasing nanoparticles on neotropical tree seedlings submitted to acclimation under full sun in the nursery.

Polymeric nanoparticles have emerged as carrier systems for molecules that release nitric oxide (NO), a free radical involved in plant stress responses. However, to date, nanoencapsulated NO donors have not been applied to plants under realistic field conditions. Here, we verified the effects of free and nanoencapsulated NO donor, S-nitroso-mercaptosuccinic acid (S-nitroso-MSA), on growth, physiological and biochemical parameters of neotropical tree seedlings kept under full sunlight in the nursery for acclimation. S-nitroso-MSA incorporation into chitosan nanoparticles partially protected the NO donor from thermal and photochemical degradation. The application of nanoencapsulated S-nitroso-MSA in the substrate favoured the growth of seedlings of Heliocarpus popayanensis, a shade-intolerant tree. In contrast, free S-nitroso-MSA or nanoparticles containing non-nitrosated mercaptosuccinic acid reduced photosynthesis and seedling growth. Seedlings of Cariniana estrellensis, a shade-tolerant tree, did not have their photosynthesis and growth affected by any formulations, despite the increase of foliar S-nitrosothiol levels mainly induced by S-nitroso-MSA-loaded nanoparticles. These results suggest that depending on the tree species, nanoencapsulated NO donors can be used to improve seedling acclimation in the nursery.

Selective Antiprotozoal Activity of Nitric Oxide-releasing Chitosan Nanoparticles Against Trypanosoma cruzi: Toxicity and Mechanisms of Action.

BACKGROUND: Chagas' disease, caused by Trypanosoma cruzi, was described for the first time over a hundred years ago. Nonetheless, clinically available drugs still lack effective and selective properties. Nitric oxide (NO) produced by activated macrophages controls the progression of disease by killing the parasite. METHODS AND RESULTS: Here, chitosan nanoparticles (CS NPs) were synthesized and mercaptosuccinic acid (MSA), the NO donor precursor, was encapsulated into CS NPs, forming MSA-CS NPs, which had hydrodynamic size of 101.0±2.535 nm. Encapsulated MSA was nitrosated forming NO donor S-nitrosomercaptosuccinic acid-containing nanoparticles (S-nitroso-MSA-CS NPs). Kinetic data revealed a sustained release of NO from the nanoparticles. S-nitroso-MSA-CS NPs inhibited epimastigote proliferation and trypomastigote viability of T. cruzi, with IC50=75.0±6.5 µg·mL-1 and EC50=25.0±5.0 µg·mL-1, respectively. Treatment of peritoneal macrophages with nanoparticles decreased the number of T. cruzi-infected cells and the average number of intracellular replicative amastigotes per infected cells. Besides, the results have showed a selective behaviour of S-nitroso-MSA-CS NPs to parasites. Morphological and biochemical changes induced by these NO-releasing nanoparticles, such as cell shrinkage, cell cycle arrest, mitochondrial membrane depolarization and phosphatidylserine exposure on cell surface indicate that epimastigotes death is associated to the apoptotic pathway. CONCLUSION: S-nitroso-MSA-CS NPs are promising nanocarriers for the treatment of Chagas's disease.