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Urban air pollution has been a global challenge world-wide. While urban vegetation or forest modelling can be useful in reducing the toxicities of the atmospheric gases by their absorption, the surge in gaseous pollutants negatively affects plant growth, thereby altering photosynthetic efficiency and harvest index. The present review analyses our current understanding of the toxic and beneficial effects of atmospheric nitrogen oxides (NO x ), hydrogen sulphide (H 2 S) and carbon monoxide (CO) on plant growth and metabolism. The atmospheric levels of these gases vary considerably due to urbanization, automobile emission, volcanic eruptions, agricultural practices and other anthropological activities. These gaseous pollutants prevalent in the atmosphere are known for their dual action (toxic or beneficiary) on plant growth, development and metabolism. NO seems to exert a specialized impact by upregulating nitrogen metabolism and reducing tropospheric ozone. High H 2 S emission in specific areas of geothermal plants, fumarolic soils and wetlands can be a limitation to air quality control. Certain shortcomings associated with the designing of field experiments, sensitivity of detection methods and simulation development are yet to be overcome to analyze the precise levels of NO, H 2 S and CO in the rhizosphere of diverse agro-climatic regions. Several laboratory-based investigations have been undertaken to assess the roles of atmospheric gases, namely NO x , CO, H 2 S, and particulate matter (PM). However, in order to enable natural and sustainable mitigation, it is essential to increase the number of field experiments in order to identify the pollutant-tolerant plants and study their interactive impact on plant growth and agriculture., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 Elsevier Ltd. All rights reserved.)

Plasma technology, which can instantaneously transform air molecules into reactive species stimulating plants, potentially contributes to developing a sustainable agricultural system with high productivity and low environmental impact. In fact, plant immunity activation by exposure to a reactive gas mainly consisting of dinitrogen pentoxide (N2O5) was recently discovered, while physiological responses to N2O5 are rarely known. Here, we demonstrate early (within 10 min) physiological responses to N2O5 gas in Arabidopsis. Exposure to N2O5 gas induced an increase in cytosolic Ca2+ concentration within seconds in directly exposed leaves, followed by systemic long-distance Ca2+-based signaling within tens of seconds. In addition, jasmonic acid (JA)-related gene expression was induced within 10 minutes, and a significant upregulation of the defense-related gene PDF1.2 was observed after 1 day of exposure to N2O5 gas. These systemic resistant responses to N2O5 were found unique among air-plasma-generated species such as ozone (O3) and nitric oxide (NO)/nitrogen dioxide (NO2). Our results provide new insights into understanding of plant physiological responses to air-derived reactive species, in addition to facilitating the development of plasma applications in agriculture., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2025 Sasaki et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Myocardial ischemia-reperfusion injury (MIRI) is an important complication in the treatment of heart failure, and its treatment has not made satisfactory progress. Nitroxyl (HNO) showed protective effects on the heart failure, however, the effect and underlying mechanism of HNO on MIRI remain largely unclear., Methods: MIRI model in this study was established to induce H9C2 cell injury through hypoxia/reoxygenation (H/R) in vitro. The cell viability was assessed by cell counting kit-8 assay. The effect of HNO on the apoptosis was detected by flow cytometry. DCFH-DA fluorescent probe method was applied to detect the level of intracellular reactive oxygen species (ROS). The morphology of mitochondria and autophagosomes were observed by transmission electron microscopy. Apoptosis, autophagy and PI3K/Akt/mTOR pathway-related proteins were detected by western blot., Results: The viability of H9C2 cells was significantly increased in the HNO group. HNO inhibited apoptosis and regulated expressions of key apoptotic protein, including Bax and Bcl-2. HNO reduced ROS levels and alleviated H/R-induced mitochondrial damage. HNO also inhibited autophagy and regulated expressions of key autophagy-related molecules, including LC3II, p62 and Beclin1. Further experiments demonstrated that the effects of HNO were mediated through upregulation of PI3K/Akt/mTOR pathway. Rapamycin reversed the inhibition of HNO on H/R-induced autophagy in H9C2 cells, which abrogated the protective effect of HNO., Conclusion: This study provided the first evidence that HNO protected H/R-induced cardiomyocytes through inhibiting autophagy via the activation PI3K/Akt/mTOR pathway., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2025 Li et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Ammonia oxidizing archaea (AOA) are among the most abundant microorganisms on earth and are known to be a major source of nitrous oxide (N 2 O) emissions, although biochemical origins of this N 2 O remain unknown. Enzymological details of AOA nitrogen metabolism are broadly unavailable. We report the recombinant expression, purification, and characterization of a multicopper oxidase, Nmar_1354, from the AOA Nitrosopumilus maritimus . We show that Nmar_1354 selectively produces nitroxyl (HNO) by coupling the oxidation of the obligate nitrification intermediate hydroxylamine (NH 2 OH) to dioxygen (O 2 ) reduction. This HNO undergoes several downstream reactions, although the major fates are production of N 2 via reaction with NH 2 OH and dimerization with itself to yield N 2 O. These results afford one plausible enzymatic origin for N 2 O release by AOA. Moreover, these results reveal a physiologically relevant enzymatic reaction for producing HNO, an enigmatic nitrogen oxide speculated to be operative in cellular signaling and in energy transduction., Competing Interests: Competing interests statement:The authors declare no competing interest.

Advanced-stage endometrial cancer patients typically receive a combination of platinum and paclitaxel chemotherapy. However, limited treatment options are available for those with recurrent disease, and there is a need to identify alternative treatment options for the advanced setting. Our goal was to evaluate the preclinical efficacy and mechanism of action of the anticancer drug Oklahoma Nitrone-007 (OKN-007) alone and in combination with carboplatin and paclitaxel in endometrial cancer. The effect of OKN-007 on the metabolic viability of endometrial cancer cells in both two- and three-dimensional (2D and 3D) cultures, as well as on clonogenic growth, in vitro was assessed. We also evaluated OKN-007 in vivo using an intraperitoneal xenograft model and targeted gene expression profiling to determine the molecular mechanism and gene expression programs altered by OKN-007. Our results showed that endometrial cancer cells were generally sensitive to OKN-007 in both 2D and 3D cultures. OKN-007 displayed a reduction in 3D spheroid and clonogenic growth. Subsequent targeted gene expression profiling revealed that OKN-007 significantly downregulated the immunosuppressive immunometabolic regulatory enzyme indolamine 2,3-dioxygenase 1 (IDO1) (-11.27-fold change) and modulated upstream inflammatory pathways that regulate IDO1 expression (interferon-gamma [IFN-γ], Janus kinase/signal transducer and activator of transcription [JAK-STAT], transforming growth factor beta [TGF-β], and nuclear factor-kappa B [NF-κB]), downstream IDO1 effector pathways (mammalian target of rapamycin [mTOR] and aryl hydrocarbon receptor [AhR]), and altered T cell signaling pathways. OKN-007 treatment reduced IDO1, sulfatase 2 (SULF2), and TGF-β protein expression in vivo and inhibited TGF-β, NF-κB, and AhR-mediated nuclear signaling in vitro. These findings indicate that OKN-007 surmounts proinflammatory, immunosuppressive, and protumorigenic pathways and is a promising approach for the effective treatment of endometrial cancer. SIGNIFICANCE STATEMENT: Women with advanced and recurrent endometrial cancer have limited therapeutic options. Oklahoma Nitrone-007 (OKN-007), which has minimal toxicity and is currently being evaluated in early-phase clinical trials for the treatment of cancer, is a potential new strategy for the treatment of endometrial cancer., Competing Interests: Conflict of interest The authors have declared a conflict of interest. R.A.T. is the holder of patents regarding the use of OKN-007 for the treatment of cancer. The remaining authors declare that they have no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Iron-sulfur cluster conversion and nitrosyl modification are involved in regulating their functions and play critical roles in signaling for biological systems. Hereby, the photo-induced dynamic process of (Me 4 N) 2 [Fe 2 S 2 (NO) 4 ] was monitored using time-resolved electron paramagnetic resonance (EPR) spectra, MS spectra and cellular imaging methods. Photo-irradiation and the solvent affect the reaction rates and products. Spectroscopic and kinetic studies have shown that the process involves at least three intermediates: spin-trapped NO free radical species with a g av at 2.040, and two other iron nitrosyl species, dinitrosyl iron units (DNICs) and mononitrosyl iron units (MNICs) with g av values at 2.031 and 2.024, respectively. Moreover, the [Fe 2 S 2 (NO) 4 ] 2- cluster could bind with ferritin and decompose gradually, and a binding state of dinitrosyl iron coordinated with Cys102 of the recombinant human heavy chain ferritin (rHuHF) was finally formed. This study provides insight into the photodynamic mechanism of nitrosyl iron - sulfur clusters to improve the understanding of physiological activity., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Reactive oxygen species generated via reperfusion cause lipid damage and induce lipid peroxidation, leading to cerebral ischemia/reperfusion injury and exacerbation of cerebral infarction. Lipid radicals are key molecules generated during lipid peroxidation. Therefore, understanding the spatiotemporal behavior of lipid radicals is important to improve the therapeutic outcomes of cerebral infarction. However, the behaviors of lipid radicals in the brain remain unclear. In this study, we aimed to evaluate the distribution of radioactivity in a transient middle cerebral artery occlusion (tMCAO) model using lipid radical detection probe [ 125 I]1 to assess the behaviors of lipid radicals after cerebral ischemia/reperfusion. The tMCAO model administered [ 125 I]1 exhibited significant differences in the timing and location of radioactivity accumulation between the ischemic and non-ischemic regions. Liquid chromatography/mass spectrometry analysis identified the lipid radical adducts formed by the reaction of 1 with the lipid radicals generated after reperfusion. More adducts were detected in the ischemic region samples than in the non-ischemic region samples. Therefore, 1 successfully detected the lipid radicals generated after cerebral ischemia/reperfusion. Overall, this study demonstrates the potential of nuclear medical imaging using radiolabeled 1 to detect the lipid radicals generated after cerebral ischemia/reperfusion. Our approach can aid in the development of new therapeutic agents scavenging lipid radicals after cerebral reperfusion by facilitating the determination of therapeutic efficacy and optimal administration period., Competing Interests: Declaration of competing interest None., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Aging is characterized as the process of functional decline in an organism from adulthood, often marked by a progressive loss of cellular function and systemic deterioration of multiple tissues. Among the numerous molecular, cellular, and systemic hallmarks associated with aging, mitochondrial dysfunction is considered one of the pivotal factors that initiates the aging process. During aging, mitochondria undergo varying degrees of damage, resulting in impaired energy production and disruption of the homeostatic regulation of mitochondrial quality control systems, which in turn affects cellular energy metabolism and results in cellular dysfunction, accelerating the aging process. AMP-activated protein kinase (AMPK) and the mechanistic target of rapamycin complex 1 (mTORC1) are two central kinase complexes responsible for sensing intracellular nutrient levels, regulating metabolic homeostasis, modulating aging and play a crucial role in maintaining the homeostatic balance of mitochondria. Our previous studies found that the novel compound tetramethylpyrazine nitrone (TBN) can protect mitochondria via the AMPK/mTOR pathway in many animal models, extending healthy lifespan through the Nrf2 signaling pathway in nematodes. Building upon this foundation, we have posited a reasonable hypothesis, TBN can improve mitochondrial function to delay aging by regulating the AMPK/mTORC1 signaling pathway. This study focuses on the C. elegans, exploring the impact and underlying mechanisms of TBN on aging and mitochondrial function (especially the mitochondrial quality control system) during the aging process. The present studies demonstrated that TBN extends lifespan of wild-type nematodes and is associated with the AMPK/mTORC1 signaling pathway. TBN elevated ATP and NAD + levels in aging nematodes while orchestrating mitochondrial biogenesis and mitophagy. Moreover, TBN was observed to significantly enhance normal activities during aging in C. elegans, such as mobility and pharyngeal pumping, concurrently impeding lipofuscin accumulation that were closely associated with AMPK and mTORC1. This study not only highlights the delayed effects of TBN on aging but also underscores its potential application in strategies aimed at improving mitochondrial function via the AMPK/mTOR pathway in C. elegans., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Inc.)

Oxidative stress is a major source of ROS-mediated damage to macromolecules, tissues, and the whole body. It is an important marker in the severe picture of pathological conditions. The discovery of free radicals in biological systems gives a "start" to studying various pathological processes related to the development and progression of many diseases. From this moment on, the enrichment of knowledge about the participation of free radicals and free-radical processes in the pathogenesis of cardiovascular, neurodegenerative, and endocrine diseases, inflammatory conditions, and infections, including COVID-19, is increasing exponentially. Excessive inflammatory responses and abnormal reactive oxygen species (ROS) levels may disrupt mitochondrial dynamics, increasing the risk of cell damage. In addition, low serum albumin levels and changes in the normal physiological balance between reduced and oxidized albumin can be a serious prerequisite for impaired antioxidant capacity of the body, worsening the condition in patients. This review presents the interrelationship between oxidative stress, inflammation, and low albumin levels, which are hallmarks of COVID-19.

Nitrosyl iron complexes are remarkably multifactorial pharmacological agents. These compounds have been proven to be particularly effective in treating cardiovascular and oncological diseases. We evaluated and compared the antioxidant activity of tetranitrosyl iron complexes (TNICs) with thiosulfate ligands and dinitrosyl iron complexes (DNICs) with glutathione (DNIC-GS) or phosphate (DNIC-PO 4 - ) ligands in hemoglobin-containing systems. The studied effects included the production of free radical intermediates during hemoglobin (Hb) oxidation by tert -butyl hydroperoxide, oxidative modification of Hb, and antioxidant properties of nitrosyl iron complexes. Measuring luminol chemiluminescence revealed that the antioxidant effect of TNICs was higher compared to DNIC-PO 4 - . DNIC-GS either did not exhibit antioxidant activity or exerted prooxidant effects at certain concentrations, which might have resulted from thiyl radical formation. TNICs and DNIC-PO 4 - efficiently protected the Hb heme group from decomposition by organic hydroperoxides. DNIC-GS did not exert any protective effects on the heme group; however, it abolished oxoferrylHb generation. TNICs inhibited the formation of Hb multimeric forms more efficiently than DNICs. Thus, TNICs had more pronounced antioxidant activity than DNICs in Hb-containing systems.

OxaD is a flavin-dependent monooxygenase (FMO) responsible for catalyzing the oxidation of an indole nitrogen atom, resulting in the formation of a nitrone. Nitrones serve as versatile intermediates in complex syntheses, including challenging reactions like cycloadditions. Traditional organic synthesis methods often yield limited results and involve environmentally harmful chemicals. Therefore, the enzymatic synthesis of nitrone-containing compounds holds promise for more sustainable industrial processes. In this study, we explored the catalytic mechanism of OxaD using a combination of steady-state and rapid-reaction kinetics, site-directed mutagenesis, spectroscopy, and structural modeling. Our investigations showed that OxaD catalyzes two oxidations of the indole nitrogen of roquefortine C, ultimately yielding roquefortine L. The reductive-half reaction analysis indicated that OxaD rapidly undergoes reduction and follows a "cautious" flavin reduction mechanism by requiring substrate binding before reduction can take place. This characteristic places OxaD in class A of the FMO family, a classification supported by a structural model featuring a single Rossmann nucleotide binding domain and a glutathione reductase fold. Furthermore, our spectroscopic analysis unveiled both enzyme-substrate and enzyme-intermediate complexes. Our analysis of the oxidative-half reaction suggests that the flavin dehydration step is the slow step in the catalytic cycle. Finally, through mutagenesis of the conserved D63 residue, we demonstrated its role in flavin motion and product oxygenation. Based on our findings, we propose a catalytic mechanism for OxaD and provide insights into the active site architecture within class A FMOs.

The effect of a promising NO donor, a binuclear nitrosyl iron complex (NIC) with 3,4-dichlorothiophenolyls [Fe 2 (SC 6 H 3 Cl 2 ) 2 (NO) 4 ], on the adenylate cyclase and soluble guanylate cyclase enzymatic systems was studied. In in vitro experiments, this complex increased the concentration of important secondary messengers, such as cAMP and cGMP. An increase of their level by 2.4 and 4.5 times, respectively, was detected at NIC concentration of 0.1 mM. The ligand of the complex, 3,4-dichlorothiophenol, produced a less pronounced effect on adenylate cyclase. It was shown that the effect of this complex on the activity of soluble guanylate cyclase was comparable to the effect of anionic nitrosyl complex with thiosulfate ligands that exhibits vasodilating and cardioprotective properties., (© 2024. Springer Science+Business Media, LLC, part of Springer Nature.)

We studied the effect of an NO donor, nitrosyl iron complex with N-ethylthiourea, on Nrf2-dependent antioxidant system activation of tumor cells in vitro. The complex increased intracellular accumulation of Nrf2 transcription factor and induced its nuclear translocation. It was shown that both heme oxygenase-1 gene and protein expression increased significantly under the influence of the complex. Nrf2 activation was accompanied by a decrease in the intracellular accumulation of proinflammatory transcription factor NF-κB p65 subunit and expression of its target genes. The cytotoxic effect of N-ethylthiourea leads to induction of Nrf2/HO-1 antioxidant response and suppression of NF-κB-dependent processes in tumor cells., (© 2024. Springer Science+Business Media, LLC, part of Springer Nature.)

Metabolism of nitric oxide (NO) donors: dinitrosyl iron complexes (DNIC), nitrosothiols (RSNO), and nitroprusside was studied on a chick embryo model. The obtained results give reason to assume that DNIC constituting the main pool of nitroso compounds in the vast majority of tissues are NO donors immediately interacting with the physiological target of NO, and other NO donors can perform this function after their transformation into DNIC. NO is released from DNIC not spontaneously, but under a joint influence of a factor destroying the complex and a target having chemical affinity for NO. A similar mechanism is apparently implicated in NO passage through the cell membrane., (© 2024. Springer Science+Business Media, LLC, part of Springer Nature.)

Oxidative stress is a major contributor to progressive neurodegenerative disease and may be a key target for the development of novel preventative and therapeutic strategies. Nitroxides have been successfully utilised to study changes in redox status (biological probes) and modulate radical-induced oxidative stress. This study investigates the efficacy of DCTEIO (5,6-dicarboxy-1,1,3,3-tetraethyllisoindolin-2-yloxyl), a stable, kinetically-persistent, nitroxide-based antioxidant, as a retinal neuroprotectant. The preservation of retinal function following an acute ischaemic/reperfusion (I/R) insult in the presence of DCTEIO was quantified by electroretinography (ERG). Inflammatory responses in retinal glia were analysed by GFAP and IBA-1 immunohistochemistry, and retinal integrity assessed by histology. A nitroxide probe combined with flow cytometry provided a rapid technique to assess oxidative stress and the mitigation offered by antioxidant compounds in cultured 661W photoreceptor cells. DCTEIO protected the retina from I/R-induced damage, maintaining retinal function. Histological analysis showed preservation of retinal integrity with reduced disruption and disorganisation of the inner and outer nuclear layers. I/R injury upregulated GFAP expression, indicative of retinal stress, which was significantly blunted by DCTEIO. The number of 'activated' microglia, particularly in the outer retina, in response to cellular stress was also significantly reduced by DCTEIO, potentially suggesting reduced inflammasome activation and cell death. DCTEIO mitigated oxidative stress in 661W retinal cell cultures, in a dose-dependent fashion. Together these findings demonstrate the potential of DCTEIO as a neuroprotective therapeutic for degenerative diseases of the CNS that involve an ROS-mediated component, including those of the retina e.g. age-related macular degeneration and glaucoma., (© 2023. The Author(s).)

Tridax procumbens (cotton buttons) is a flowering plant with a medicinal reputation for treating infections, wounds, diabetes, and liver and kidney diseases. The present research was conducted to evaluate the possible protective effects of the T. procumbens methanolic extract (TPME) on an experimentally induced type 2 diabetes rat model. Wistar rats with streptozotocin (STZ)-induced diabetes were randomly allocated into five groups of five animals each, viz., a normal glycemic group (I), diabetic rats receiving distilled water group (II), diabetic rats with 150 (III) and 300 mg/kg of TPME (IV) groups, and diabetic rats with 100 mg/kg metformin group (V). All treatments were administered for 21 consecutive days through oral gavage. Results: Administration of the T. procumbens extract to diabetic rats significantly restored alterations in levels of fasting blood glucose (FBG), body weight loss, serum and pancreatic insulin levels, and pancreatic histology. Furthermore, T. procumbens significantly attenuated the dyslipidemia (increased cholesterol, low-density lipoprotein-cholesterol (LDL-C), triglycerides, and high-density lipoprotein (HDL) in diabetic rats), serum biochemical alterations (alanine transaminase (ALT), aspartate transaminase (AST), alanine phosphatase (ALP), blood urea nitrogen (BUN), creatinine, uric acid, and urea) and full blood count distortion in rats with STZ-induced diabetes. The TPME also improved the antioxidant status as evidenced by increased superoxide dismutase (SOD), catalase (CAT), glutathione (GSH), and decreased malondialdehyde (MDA); and decreased levels of cholinesterases (acetylcholinesterase (AChE) and butyrylcholinesterase (BChE)), and proinflammatory mediators including nuclear factor (NF)-κB, cyclooxygenase (COX)- 2, and nitrogen oxide (NOx) in the brain of rats with STZ-induced diabetes compared to rats with STZ-induced diabetes that received distilled water. However, TPME treatment failed to attenuate the elevated monoamine oxidases and decreased dopamine levels in the brain of rats with STZ-induced diabetes. Extract characterization by liquid chromatography mass spectrometry (LC-MS) identified isorhamnetin (retention time (RT)= 3.69 min, 8.8%), bixin (RT: 25.06 min, 4.72%), and lupeol (RT: 25.25 min, 2.88%) as the three most abundant bioactive compounds that could be responsible for the bioactivity of the plant. In conclusion, the TPME can be considered a promising alternative therapeutic option for managing diabetic complications owing to its antidiabetic, antihyperlipidemic, antioxidant, and anti-inflammatory effects in rats with STZ-prompted diabetes., Competing Interests: Conflict of interest statement The authors declare no conflict of interest., (Copyright © 2022 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

Polarity reversal is one of the effective strategies to rapidly start up denitrifying BESs，but the long-term performances of the denitrifying BESs operated under polarity reversal receive little attention. This study investigated the effects of periodic polarity reversal (PPR) and polarity reversal once only (PRO) on the long-term performances of denitrifying BESs. Repeatable oxidative and reductive currents were observed in the BESs obtained by PPR (PPR-BESs). The peak reductive currents of the PPR-BESs reached 0.95 A/m 2 , and nitrate was mainly removed by dissimilatory nitrate reduction to ammonium pathway with removal rates higher than 95 %. In contrast, the peak reductive currents of the BESs obtained by PRO (PRO-BESs) progressively decreased from 1.01 A/m 2 to 0.12 A/m 2 . The nitrate removal rates of the PRO-BESs were <50 %, and the product of nitrate reduction turned to N 2 instead of ammonium. 16S rDNA sequencing and metatranscriptomic analysis revealed that Geobacter capable of bidirectional extracellular electron transfer (EET) and Afipia capable of autotrophic growth were the dominant genera in the two types of BESs. Outer membrane cytochrome c and formate dehydrogenase were potentially involved in the cathodic electron uptake. These findings contribute to a better understanding of the EET mechanisms of electroautotrophic denitrifiers., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Nitrate promotes anodic denitrification and fasts organic matter removal in microbial fuel cells (MFCs). However, it suffers from poor total nitrogen (TN) removal and current recovery. In this study, some novel electroactive nitrifying/denitrifying bacteria (ENDB) were introduced in a single chambered air-cathode MFC to investigate the performance of this device and the microbial community shift by adding nitrate. Results showed a similar disturbance in current output by adding nitrate during a short-term operation. However, a stable and reproducible current increase was achieved in the continuous experiment. A maximum current of 0.76 A m -3 and a maximum TN removal of >99 % were accomplished. The corresponding corrected coulombic efficiency was approximately 18 %. Under repeatable batches, a sharp decrease in chemical oxygen demand (COD) with feeding nitrate confirmed the temporary competition on electron donors through heterotrophic denitrification. The later current increase and nitrite detection occurring without metabolized COD could be considered evidence of electroactive anodic nitrification. The ENDB biofilm successfully coupled mixotrophic denitrification and electroactive anodic nitrification. It eventually promoted TN removal. In the process, genera Pseudoxanthomonas, Thauera, and Pseudomonas were enriched in the anodic ENDB biofilms. Cyclic voltammetry data confirmed the promotion of the electron transfer process by biofilms. The bacterial function predication revealed that the genes related to nitrogen removal and electron transfer were upregulated. Therefore, mixotrophic denitrification and electroactive anodic nitrification processes facilitated power recovery with the high efficiency of pollutant removal, finally ensuring water body security., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Nitrate is a common pollutant in the aquatic environment. Denitrification and dissimilatory nitrate reduction to ammonium (DNRA) are the main reduction processes of nitrate. In the relatively closed sediment environment, the competitive interaction of these two nitrate reduction determines whether the ecosystem removes or retains nitrogen. In the process of NO 3 - -N bioreduction, Magnetite, which is a common mineral present in soil and other sediments can play a crucial role. However, it is still not clear whether magnetite promotes or inhibits NO 3 - -N bioreduction. In this paper, the effect of magnetite on NO 3 - -N bioreduction was studied by batch experiments. The results show that magnetite can increase the NO 3 - -N reduction rate by 1.48 %, and can inhibit the DNRA process at the beginning of the reaction and then promote the DNRA process. Magnetite changed the microbial community structure in our experiment systems. The relative abundance of Sphingomonas, which mainly exists in a high carbon and low nitrogen environment, increased under sufficient carbon source conditions. The relative abundance of Fe-oxidizing and NO 3 - -N reducing bacteria, such as Flavobacterium, increased in the absence of carbon sources but in the presence of magnetite. In addition, magnetite can significantly increase activity of the microbial electron transport system (ETS). the added microbial electronic activity of magnetite increased nearly two-fold under the same experiment conditions. The acid produced by the metabolisms of Pseudomonas and Acinetobacter further promotes the dissolution of magnetite, thus increasing the concentration of Fe (II) in the system, which is beneficial to autotrophic denitrifying bacteria and promote the reduction of NO 3 - -N. These findings can enhance our understanding of the interaction mechanism between iron minerals and nitrate reducing bacteria during nitrate reduction under natural conditions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022. Published by Elsevier B.V.)

Flue gas not only contains carbon dioxide (CO 2 ) but also air pollutants (sulfur oxides (SO x ) and nitrogen oxides (NO x )). The effective utilization of flue gas could help us to reduce the cost of microalgal biomass production. This study assessed and explored the utilization of flue gas for the absorption characteristics of different components and their biological effect in microalgal culture systems. In abiotic absorption experiments, the absorptivity of CO 2 was reduced by a maximum of 3.1%, and the concentration of the available carbon source in the culture medium was decreased by 6.7% when sulfur dioxide (SO 2 , at 100 mg/m 3 ) was presented in the flue gas. Meanwhile, the presence of oxygen (O 2 , at 4%) in the flue gas improved the absorptivity of nitric oxide (NO). When Scenedesmus dimorphus was cultured using bisulfites and nitrites (at 10 mmol/L and 8 mmol/L, respectively) as the sulfur and nitrogen sources, SO x and NO x in the flue gas did not significantly affect growth of microalgal cells and the carbohydrate, lipid, and protein content. The consumption rates of nutrient elements were calculated, which could provide an adjustment strategy for the initial gas source when culturing microalgae with the flue gas. This study indicates that the flue gas used for microalgal culture should be partially desulfurized, so that the SO x and CO 2 concentrations can optimize growth of microalgal cells, while the denitrification might not be needed since the flue gas can be oxidized to utilize the NO. KEY POINTS: • The concentration of the available carbon source in the culture medium was decreased when SO 2 was presented in the flue gas, and the presence of O 2 in the flue gas improved the absorptivity of NO. • An adjustment strategy for the initial gas source when culturing microalgae with the flue gas was firstly proposed. • For flue gas containing 10% CO 2 and 60 mg/m 3 of SO 2 , growth of Scenedesmus dimorphus showed no difference in cell growth in normal culture conditions., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Nitric oxide (NO) stimulates mitochondrial biogenesis in skeletal muscle. However, NO metabolism is disrupted in individuals with type 2 diabetes mellitus (T2DM) potentially contributing to their decreased cardiorespiratory fitness (i.e., VO2max) and skeletal muscle oxidative capacity. We used a randomized, double-blind, placebo-controlled, 8-week trial with beetroot juice containing nitrate (NO3−) and nitrite (NO2−) (250 mg and 20 mg/day) to test potential benefits on VO2max and skeletal muscle oxidative capacity in T2DM. T2DM (N = 36, Age = 59 ± 9 years; BMI = 31.9 ± 5.0 kg/m2) and age- and BMI-matched non-diabetic controls (N = 15, Age = 60 ± 9 years; BMI = 29.5 ± 4.6 kg/m2) were studied. Mitochondrial respiratory capacity was assessed in muscle biopsies from a subgroup of T2DM and controls (N = 19 and N = 10, respectively). At baseline, T2DM had higher plasma NO3− (100%; p < 0.001) and lower plasma NO2− levels (−46.8%; p < 0.0001) than controls. VO2max was lower in T2DM (−26.4%; p < 0.001), as was maximal carbohydrate- and fatty acid-supported oxygen consumption in permeabilized muscle fibers (−26.1% and −25.5%, respectively; p < 0.05). NO3−/NO2− supplementation increased VO2max (5.3%; p < 0.01). Further, circulating NO2−, but not NO3−, positively correlated with VO2max after supplementation (R2= 0.40; p < 0.05). Within the NO3−/NO2− group, 42% of subjects presented improvements in both carbohydrate- and fatty acid-supported oxygen consumption in skeletal muscle (vs. 0% in placebo; p < 0.05). VO2max improvements in these individuals tended to be larger than in the rest of the NO3−/NO2− group (1.21 ± 0.51 mL/(kg*min) vs. 0.31 ± 0.10 mL/(kg*min); p = 0.09). NO3−/NO2− supplementation increases VO2max in T2DM individuals and improvements in skeletal muscle oxidative capacity appear to occur in those with more pronounced increases in VO2max.

Nitrate pollution has become a worldwide problem. In this study, we remove nitrate from water by electrodialysis ion-exchange membrane bioreactor (EDIMB) and enabling simultaneous nitrate enrichment and denitrification. In this reactor, nitrate migrated from the water chamber to the biological chamber via electrodialysis and was degraded by microorganisms. The effects of voltage and biomass concentration on the reactor performance were examined, and the kinetics data of the water chamber and biological chamber were fitted. The experimental results showed that the migration of nitrate in the water chamber conformed to the first-order model, and the constructed zero-Michaelis-Menten model described changes in nitrate concentration in the biological chamber. Furthermore, when the inflow nitrate concentration was 40 mg N/L, 5 V was the best voltage, and 3.00 g VSS/L was the best biomass concentration. The nitrate removal rate in the water chamber was 98.94%, and there was no accumulation of nitrate or nitrite in the biological chamber. Compared with traditional ED processes, the nitrate removal efficiency was 8.86% higher, and the current efficiency was 22.14% higher. The total organic carbon (TOC) of the water chamber was only 1.43 mg C/L, which proves that the structure of the EDIMB confined the denitrifying bacteria and organic carbon donors in the biological chamber and avoided secondary pollution in the water chamber. Microbial community analysis showed that Thauera (66.06%) was the dominant bacterium in the EDIMB system, and Azoarcus (9.81%) was a minor denitrifying genus., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Nitric oxide (NO) is an active and critical nitrogen oxide in the microbe-driven nitrogen biogeochemical cycle, and is of great interest to medicine and the biological sciences. As a gas molecule prior to oxygen, NO respiration represents an early form of energy generation via various reactions in prokaryotes. Major enzymes for endogenous NO formation known to date include two types of nitrite reductases in denitrification, hydroxylamine oxidoreductase in ammonia oxidation, and NO synthases (NOSs). While the former two play critical roles in shaping electron transport pathways in bacteria, NOSs are intracellular enzymes catalyzing metabolism of certain amino acids and have been extensively studied in mammals. NO interacts with numerous cellular targets, most of which are redox-active proteins. Doing so, NO plays harmful and beneficial roles by affecting diverse biological processes within bacterial physiology. Here, we discuss recent advances in the field, including NO-forming enzymes, the molecular mechanisms by which these enzymes function, physiological roles of bacterial NOSs, and regulation of NO homeostasis in bacteria.

Hypertension has become one of the important diseases harmful to human health. In China, Qingda granule (QDG) has been used to treat hypertension for decades. Previous studies by our team have shown that oxidative stress may be one of the pathways through which QDG inhibits hypertension-induced organs injury. However, the specific molecular mechanism of its anti-hypotension and renal oxidative stress response were unclearly. This study investigated QDG's potential protective mechanism against hypertension-induced renal injury. Mice were infused with Angiotensin Ⅱ (Ang Ⅱ, 500 ng/kg/min) or equivalent saline solution (Control) and administered oral QDG (1.145 g/kg/day) or saline for four weeks. QDG treatment mitigated the elevated blood pressure and reduced renal pathological changes induced by Ang Ⅱ. As per the RNA sequencing results, QDG affects oxidative stress signaling. In agreement with these findings, QDG significantly attenuated the Ang Ⅱ-induced increase in Nitrogen oxides 1 (NOX1) and reactive oxygen species and the decrease in superoxide dismutase in renal tissue. Additionally, QDG significantly inhibited Interleukin 6 (IL-6), Tumor necrosis factor α (TNF-α), and Interleukin 1β (IL-1β) expression in renal tissues and blocked the phosphorylation of P65 (NF-κB subunit) and IκB. These results were confirmed in vitro. Overall, QDG reduced Ang Ⅱ-induced elevated blood pressure and renal injury by inhibiting oxidative stress and inflammation caused by NOX1 and NF-κB pathways. The results of this study provide an experimental basis for the clinical application of QDG, and to open up a new direction for the clinical treatment of hypertension., Competing Interests: Conflicts of interest The authors declare that they have no conflicts of interest., (Copyright © 2022 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

Nitrate is the preferred nitrogen source for plants and plays an important role in plant growth and development. Under various soil stresses, plants reallocate nitrate to roots to promote stress tolerance through the ethylene-ethylene response factors (ERFs)-nitrate transporter (NRT) signaling module. As a light signal, ultraviolet B (UV-B) also stimulates the production of ethylene. However, whether UV-B regulates nitrate reallocation in plants via ethylene remains unknown. Here, we found that UV-B-induced expression of ERF1B , ORA59 , ERF104 , and NRT1.8 in both Arabidopsis shoots and roots as well as nitrate reallocation from hypocotyls to leaves and roots were impaired in ethylene signaling mutants for Ethylene Insensitive2 (EIN2) and EIN3. UV-B-induced NRT1.8 expression and nitrate reallocation to leaves and roots were also inhibited in the triple mutants for ERF1B , ORA59 , and ERF104 . Deletion of NRT1.8 impaired UV-B-induced nitrate reallocation to both leaves and roots. Furthermore, UV-B promoted ethylene release in both shoots and roots by enhancing the gene expression and enzymatic activities of ethylene biosynthetic enzymes only in shoots. These results show that ethylene acts as a local and systemic signal to mediate UV-B-induced nitrate reallocation from Arabidopsis hypocotyls to both leaves and roots via regulating the gene expression of the ERFs-NRT1.8 signaling module.

The human microbiome comprises ∼10 13 -10 14 microbial cells which form a symbiotic relationship with the host and play a critical role in the regulation of human metabolism. In the oral cavity, several species of bacteria are capable of reducing nitrate to nitrite; a key precursor of the signaling molecule nitric oxide. Nitric oxide has myriad physiological functions, which include the maintenance of cardiovascular homeostasis and the regulation of acute and chronic responses to exercise. This article provides a brief narrative review of the research that has explored how diversity and plasticity of the oral microbiome influences nitric oxide bioavailability and related physiological outcomes. There is unequivocal evidence that dysbiosis (e.g. through disease) or disruption (e.g. by use of antiseptic mouthwash or antibiotics) of the oral microbiota will suppress nitric oxide production via the nitrate-nitrite-nitric oxide pathway and negatively impact blood pressure. Conversely, there is preliminary evidence to suggest that proliferation of nitrate-reducing bacteria via the diet or targeted probiotics can augment nitric oxide production and improve markers of oral health. Despite this, it is yet to be established whether purposefully altering the oral microbiome can have a meaningful impact on exercise performance. Future research should determine whether alterations to the composition and metabolic activity of bacteria in the mouth influence the acute responses to exercise and the physiological adaptations to exercise training., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Twelve sampling sites from two basins of Lake Chaohu were studied seasonally from June 2020 to April 2021 in Hefei City (China) to better understand the effect of organic carbon (C) quantity and composition on nitrate (NO 3 - -N) reduction pathways. Serious algal bloom in the west basin of Lake Chaohu (WLC) resulted in higher organic C accumulation and NO 3 - -N deficiency in interstitial water compared to the east basin of Lake Chaohu (ELC), jointly leading to a high C/NO 3 - -N ratio. This triggered dissimilatory nitrate reduction to ammonium (DNRA) over denitrification in terms of higher DNRA rate, nitrogen retaining index (NRI), and nrfA gene abundance mediating DNRA. Furthermore, high oxygen-alkyl C and abundance of functional genes mediating labile organic C decomposition and DNRA suggested that the alkyl carbon-oxygen bond was responsible for DNRA induction. Different bacterial community composition and diversity involved in C and nitrogen (N) metabolism in two basins indicated that bacteria in sediments of WLC were more active in NO 3 - -N reduction. Spearman correlation analysis showed that the less represented genera, such as Thiobacillus and Clostridium, were positively correlated with both organic C and NO 3 - -N reduction rates, respectively. Hence, organic C composition could affect NO 3 - -N reduction function by shaping the specific bacterial community., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The role of root phenes in nitrogen (N) acquisition and biomass production was evaluated in 10 contrasting natural accessions of Arabidopsis thaliana L. Seedlings were grown on vertical agar plates with two different nitrate supplies. The low N treatment increased the root to shoot biomass ratio and promoted the proliferation of lateral roots and root hairs. The cost of a larger root system did not impact shoot biomass. Greater biomass production could be achieved through increased root length or through specific root hair characteristics. A greater number of root hairs may provide a low-resistance pathway under elevated N conditions, while root hair length may enhance root zone exploration under low N conditions. The variability of N uptake and the expression levels of genes encoding nitrate transporters were measured. A positive correlation was found between root system size and high-affinity nitrate uptake, emphasizing the benefits of an exploratory root organ in N acquisition. The expression levels of NRT1.2/NPF4.6, NRT2.2, and NRT1.5/NPF7.3 negatively correlated with some root morphological traits. Such basic knowledge in Arabidopsis demonstrates the importance of root phenes to improve N acquisition and paves the way to design eudicot ideotypes., (© The Author(s) 2022. Published by Oxford University Press on behalf of Sleep Research Society. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

In this study, we discussed the relationship between Entermorpha linza polysaccharide (EP) and Bacillus subtilis, which can transform nitrate. A sole carbon source experiment showed that Bacillus subtilis could utilize EP, and the bacterial density was maximally increased by 54.43% in the EP groups. The results of reducing sugar determination proved the secretion of polysaccharide-degrading enzymes. Scanning electron microscopy (SEM) showed that the EP groups had fewer spores and shrunken bacteria, indicating that EP could improve the growth environment and maintain bacterial integrity. Additionally, the ratios of periplasmic nitrate reductase (NAP), nitrite reductase (NIR), and dissimilatory nitrate reductase (D-NRase) in the EP groups were maximally increased by 107.22%, 84.70% and 36.10%, respectively. Transcriptome analysis further confirmed the above mentioned results. For example, the high expression of quorum sensing genes indicated that EP groups had higher bacterial density. Moreover, the high expression of antioxidant genes in the EP groups may be related to morphological integrity. Our study provides a basis for further discussion of the mechanism., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Pseudomonas aeruginosa is an opportunistic pathogen and well characterized biofilm former. P. aeruginosa forms strong oxygen gradients inside biofilms due to rapid oxygen respiration in the top layers and the poor solubility of oxygen coupled with diffusion limited transport. Transcriptomic evidence from in vitro and ex vivo sampling suggests that denitrification is occurring in biofilms in ostensibly oxic environments. It is hypothesized that in the presence of nitrate there is stratification with aerobic respiration occurring in the outer oxic layer and denitrification in the lower anoxic zone. We used submerged agar colony biofilms grown from mucoid (FRD1) and non-mucoid (PAO1) strains to simultaneously measure depth microprofiles of oxygen and nitrous oxide in the same colony with microelectrodes. Oxygen respiration occurred at the top of the colony as expected but denitrification occurred throughout the entire depth, even in the oxic region. Local denitrification rates were highly variable suggesting heterogenous metabolic activity within the colony. We also assessed the short-term influence of tobramycin on aerobic respiration within a PAO1 colony. Although there was an immediate reduction in respiration it was never completely arrested over a 2 h period. On tobramycin removal the oxygen gradient steadily reestablished, demonstrating immediate recovery of metabolic activity., (© 2022. The Author(s).)

Nitrous oxide (N 2 O) is a powerful greenhouse gas that contributes to climate change. Denitrification is one of the largest sources of N 2 O in soils. The soybean endosymbiont Bradyrhizobium diazoefficiens is a model for rhizobial denitrification studies since, in addition to fixing N 2 , it has the ability to grow anaerobically under free-living conditions by reducing nitrate from the medium through the complete denitrification pathway. This bacterium contains a periplasmic nitrate reductase (Nap), a copper (Cu)-containing nitrite reductase (NirK), a c -type nitric oxide reductase (cNor), and a Cu-dependent nitrous oxide reductase (Nos) encoded by the napEDABC, nirK, norCBQD and nosRZDFYLX genes, respectively. In this work, an integrated study of the role of Cu in B. diazoefficiens denitrification has been performed. A notable reduction in nirK , nor, and nos gene expression observed under Cu limitation was correlated with a significant decrease in NirK, NorC and NosZ protein levels and activities. Meanwhile, nap expression was not affected by Cu, but a remarkable depletion in Nap activity was found, presumably due to an inhibitory effect of nitrite accumulated under Cu-limiting conditions. Interestingly, a post-transcriptional regulation by increasing Nap and NirK activities, as well as NorC and NosZ protein levels, was observed in response to high Cu. Our results demonstrate, for the first time, the role of Cu in transcriptional and post-transcriptional control of B. diazoefficiens denitrification. Thus, this study will contribute by proposing useful strategies for reducing N 2 O emissions from agricultural soils.

S -nitrosothiols are labile thiol-NO adducts formed in vivo primarily by metalloproteins such as NO synthase, ceruloplasmin, and hemoglobin. Abnormal S -nitrosothiol synthesis and catabolism contribute to many diseases, ranging from asthma to septic shock. Current methods for quantifying S -nitrosothiols in vivo are suboptimal. Samples need to be removed from the body for analysis, and the S -nitrosothiols can be broken down during ex vivo processing. Here, we have developed a noninvasive device to measure mammalian tissue S -nitrosothiols in situ non-invasively using ultraviolet (UV) light, which causes NO release in proportion to the S -nitrosothiol concentration. We validated the assay in vitro; then, we applied it to measure S -nitrosothiols in vivo in rats and in humans. The method was sensitive to 0.5 µM, specific (did not detect other nitrogen oxides), and was reproducible in rats and in humans. This noninvasive approach to S -nitrosothiol measurements may be applicable for use in human diseases.

Neuronal nitric oxide synthase (nNOS) catalyzes single-electron reduction of quinones (Q), nitroaromatic compounds (ArNO 2 ) and aromatic N -oxides (ArN → O), and is partly responsible for their oxidative stress-type cytotoxicity. In order to expand a limited knowledge on the enzymatic mechanisms of these processes, we aimed to disclose the specific features of nNOS in the reduction of such xenobiotics. In the absence or presence of calmodulin (CAM), the reactivity of Q and ArN → O increases with their single-electron reduction midpoint potential ( E 1 7 ). ArNO 2 form a series with lower reactivity. The calculations according to an "outer-sphere" electron transfer model show that the binding of CAM decreases the electron transfer distance from FMNH 2 to quinone by 1-2 Å. The effects of ionic strength point to the interaction of oxidants with a negatively charged protein domain close to FMN, and to an increase in accessibility of the active center induced by high ionic strength. The multiple turnover experiments of nNOS show that, in parallel with reduced FAD-FMN, duroquinone reoxidizes the reduced heme, in particular its Fe 2+ -NO form. This finding may help to design the heme-targeted bioreductively activated agents and contribute to the understanding of the role of P-450-type heme proteins in the bioreduction of quinones and other prooxidant xenobiotics.

In this article we minutely discuss the so-called "oxidative" mechanism of mononuclear form of dinitrosyl iron complexes (M-DNICs) formations proposed by the author. M-DNICs are proposed to be formed from their building material-neutral NO molecules, Fe 2+ ions and anionic non-thiol (L - ) and thiol (RS - ) ligands based on the disproportionation reaction of NO molecules binding with divalent ion irons in pairs. Then a protonated form of nitroxyl anion (NO - ) appearing in the reaction is released from this group and a neutral NO molecule is included instead. As a result, M-DNICs are produced. Their resonance structure is described as [(L - ) 2 Fe 2+ (NO)(NO + )], in which nitrosyl ligands are represented by NO molecules and nitrosonium cations in equal proportions. Binding of hydroxyl ions with the latter causes conversion of these cations into nitrite anions at neutral pH values and therefore transformation of DNICs into the corresponding high-spin mononitrosyl iron complexes (MNICs) with the resonance structure described as [(L - ) 2 Fe 2+ (NO)]. In case of replacing L - by thiol-containing ligands, which are characterized by high π-donor activity, electron density transferred from sulfur atoms to iron-dinitrosyl groups neutralizes the positive charge on nitrosonium cations, which prevents their hydrolysis, ensuring relatively a high stability of the corresponding M-DNICs with the resonance structure [(RS - ) 2 Fe 2+ (NO, NO + )]. Therefore, M-DNICs with thiol-containing ligands, as well as their binuclear analogs (B-DNICs, respective resonance structure [(RS - ) 2 Fe 2+ 2 (NO, NO + ) 2 ]), can serve donors of both NO and NO + . Experiments with solutions of B-DNICs with glutathione or N-acetyl-L-cysteine (B-DNIC-GSH or B-DNIC-NAC) showed that these complexes release both NO and NO + in case of decomposition in the presence of acid or after oxidation of thiol-containing ligands in them. The level of released NO was measured via optical absorption intensity of NO in the gaseous phase, while the number of released nitrosonium cations was determined based on their inclusion in S-nitrosothiols or their conversion into nitrite anions. Biomedical research showed the ability of DNICs with thiol-containing ligands to be donors of NO and NO + and produce various biological effects on living organisms. At the same time, NO molecules released from DNICs usually have a positive and regulatory effect on organisms, while nitrosonium cations have a negative and cytotoxic effect.

In a relatively isolated system of avian embryo, the metabolism of NO, a component of the dinitrosyl iron complexes (DNIC), the main NO donor in most tissues, depends on the ligands that make up the complex. This fact corroborates the earlier hypothesis that these ligands perform a regulatory function in NO metabolism. It is also shown that nitrite injected into the embryo is not oxidized to nitrate like NO in DNIC, but is accumulated outside the amniotic sac. Normally, nitrite is present in an embryo in trace amounts. These facts suggest that NO in the embryo is transferred from the donor molecule to a target in the embryo tissues further transformed with minimum oxidation to nitrite., (© 2021. Springer Science+Business Media, LLC, part of Springer Nature.)

Endothelial hemoglobin (Hb)α regulates endothelial nitric oxide synthase (eNOS) biochemistry. We hypothesized that Hb could also be expressed and biochemically active in the ciliated human airway epithelium. Primary human airway epithelial cells, cultured at air-liquid interface (ALI), were obtained by clinical airway brushings or from explanted lungs. Human airway Hb mRNA data were from publically available databases; or from RT-PCR. Hb proteins were identified by immunoprecipitation, immunoblot, immunohistochemistry, immunofluorescence and liquid chromatography- mass spectrometry. Viral vectors were used to alter Hbβ expression. Heme and nitrogen oxides were measured colorimetrically. Hb mRNA was expressed in human ciliated epithelial cells. Heme proteins (Hbα, β, and δ) were detected in ALI cultures by several methods. Higher levels of airway epithelial Hbβ gene expression were associated with lower FEV 1 in asthma. Both Hbβ knockdown and overexpression affected cell morphology. Hbβ and eNOS were apically colocalized. Binding heme with CO decreased extracellular accumulation of nitrogen oxides. Human airway epithelial cells express Hb. Higher levels of Hbβ gene expression were associated with airflow obstruction. Hbβ and eNOS were colocalized in ciliated cells, and heme affected oxidation of the NOS product. Epithelial Hb expression may be relevant to human airways diseases., (© 2021. The Author(s).)

Nitric oxide (NO) is a short-lived signaling molecule that plays a pivotal role in cardiovascular system. Organic nitrates represent a class of NO-donating drugs for treating coronary artery diseases, acting through the vasodilation of systemic vasculature that often leads to adverse effects. Herein, we design a nitrate-functionalized patch, wherein the nitrate pharmacological functional groups are covalently bound to biodegradable polymers, thus transforming small-molecule drugs into therapeutic biomaterials. When implanted onto the myocardium, the patch releases NO locally through a stepwise biotransformation, and NO generation is remarkably enhanced in infarcted myocardium because of the ischemic microenvironment, which gives rise to mitochondrial-targeted cardioprotection as well as enhanced cardiac repair. The therapeutic efficacy is further confirmed in a clinically relevant porcine model of myocardial infarction. All these results support the translational potential of this functional patch for treating ischemic heart disease by therapeutic mechanisms different from conventional organic nitrate drugs., (© 2021. The Author(s).)

Aims: The aim of this study was to obtain cold-adapted denitrifying fungi that could be used for bioaugmentation in woodchip bioreactors to remove nitrate from agricultural subsurface drainage water., Methods and Results: We isolated a total of 91 nitrate-reducing fungal strains belonging to Ascomycota and Mucoromycota from agricultural soil and a woodchip bioreactor under relatively cold conditions (5 and 15°C). When these strains were incubated with 15 N-labelled nitrate, 29 N 2 was frequently produced, suggesting the occurrence of co-denitrification (microbially mediated nitrosation). Two strains also produced 30 N 2 , indicating their ability to reduce N 2 O. Of the 91 nitrate-reducing fungal strains, fungal nitrite reductase gene (nirK) and cytochrome P450 nitric oxide reductase gene (p450nor) were detected by PCR in 34 (37%) and 11 (12%) strains, respectively. Eight strains possessed both nirK and p450nor, further verifying their denitrification capability. In addition, most strains degraded cellulose under denitrification condition., Conclusions: Diverse nitrate-reducing fungi were isolated from soil and a woodchip bioreactor. These fungi reduced nitrate to gaseous N forms at relatively low temperatures. These cold-adapted, cellulose-degrading and nitrate-reducing fungi could support themselves and other denitrifiers in woodchip bioreactors., Significance and Impact of the Study: The cold-adapted, cellulose-degrading and nitrate-reducing fungi isolated in this study could be useful to enhance nitrate removal in woodchip bioreactors under low-temperature conditions., (© 2020 The Society for Applied Microbiology.)

Nitric oxide (NO) reductase from the fungus Fusarium oxysporum is a P450-type enzyme (P450nor) that catalyzes the reduction of NO to nitrous oxide (N 2 O) in the global nitrogen cycle. In this enzymatic reaction, the heme-bound NO is activated by the direct hydride transfer from NADH to generate a short-lived intermediate ( I ), a key state to promote N-N bond formation and N-O bond cleavage. This study applied time-resolved (TR) techniques in conjunction with photolabile-caged NO to gain direct experimental results for the characterization of the coordination and electronic structures of I TR freeze-trap crystallography using an X-ray free electron laser (XFEL) reveals highly bent Fe-NO coordination in I , with an elongated Fe-NO bond length (Fe-NO = 1.91 Å, Fe-N-O = 138°) in the absence of NAD + TR-infrared (IR) spectroscopy detects the formation of I with an N-O stretching frequency of 1,290 cm -1 upon hydride transfer from NADH to the Fe 3+ -NO enzyme via the dissociation of NAD + from a transient state, with an N-O stretching of 1,330 cm -1 and a lifetime of ca. 16 ms. Quantum mechanics/molecular mechanics calculations, based on these crystallographic and IR spectroscopic results, demonstrate that the electronic structure of I is characterized by a singly protonated Fe 3+ -NHO •- radical. The current findings provide conclusive evidence for the N 2 O generation mechanism via a radical-radical coupling of the heme nitroxyl complex with the second NO molecule., Competing Interests: The authors declare no competing interest.

Intracellular redox imbalance in endothelial cells (EC) can lead to endothelial dysfunction, which underpins cardiovascular diseases (CVD). The acute phase serum amyloid A (SAA) elicits inflammation through stimulating production of reactive oxygen species (ROS). The cyclic nitroxide 4-MethoxyTEMPO (4-MetT) is a superoxide dismutase mimetic that suppresses oxidant formation and inflammation. The aim of this study was to investigate whether 4-MetT inhibits SAA-mediated activation of cultured primary human aortic EC (HAEC). Co-incubating cells with 4-MetT inhibited SAA-mediated increases in adhesion molecules (VCAM-1, ICAM-1, E-selectin, and JAM-C). Pre-treatment of cells with 4-MetT mitigated SAA-mediated increases in transcriptionally activated NF-κB-p65 and P120 Catenin (a stabilizer of Cadherin expression). Mitochondrial respiration and ROS generation (mtROS) were adversely affected by SAA with decreased respiratory reserve capacity, elevated maximal respiration and proton leakage all characteristic of SAA-treated HAEC. This altered respiration manifested as a loss of mitochondrial membrane potential (confirmed by a decrease in TMRM fluorescence), and increased mtROS production as assessed with MitoSox Red. These SAA-linked impacts on mitochondria were mitigated by 4-MetT resulting in restoration of HAEC nitric oxide bioavailability as confirmed by assessing cyclic guanosine monophosphate (cGMP) levels. Thus, 4-MetT ameliorates SAA-mediated endothelial dysfunction through normalising EC redox homeostasis. Subject to further validation in in vivo settings; these outcomes suggest its potential as a therapeutic in the setting of cardiovascular pathologies where elevated SAA and endothelial dysfunction is linked to enhanced CVD.

The circadian clock at the hypothalamic suprachiasmatic nucleus (SCN) entrains output rhythms to 24-h light cycles. To entrain by phase-advances, light signaling at the end of subjective night (circadian time 18, CT18) requires free radical nitric oxide (NO•) binding to soluble guanylate cyclase (sGC) heme group, activating the cyclic guanosine monophosphate (cGMP)-dependent protein kinase (PKG). Phase-delays at CT14 seem to be independent of NO•, whose redox-related species were yet to be investigated. Here, the one-electron reduction of NO• nitroxyl was pharmacologically delivered by Angeli's salt (AS) donor to assess its modulation on phase-resetting of locomotor rhythms in hamsters. Intracerebroventricular AS generated nitroxyl at the SCN, promoting phase-delays at CT14, but potentiated light-induced phase-advances at CT18. Glutathione/glutathione disulfide (GSH/GSSG) couple measured in SCN homogenates showed higher values at CT14 (i.e., more reduced) than at CT18 (oxidized). In addition, administration of antioxidants N-acetylcysteine (NAC) and GSH induced delays per se at CT14 but did not affect light-induced advances at CT18. Thus, the relative of NO• nitroxyl generates phase-delays in a reductive SCN environment, while an oxidative favors photic-advances. These data suggest that circadian phase-locking mechanisms should include redox SCN environment, generating relatives of NO•, as well as coupling with the molecular oscillator.

A new Ni(ii) complex [NiL] (complex 1) of an asymmetrically di-condensed N 2 O 3 donor Schiff base ligand, N-salicylidene-N'-3-methoxysalicylidene-1,3-propanediamine (H 2 L), has been synthesized and utilized for the synthesis of three heterometallic complexes, [(NiL) 2 Mn(NCS) 2 (CH 3 OH) 2 ]·CH 3 OH (2) [(NiL) 2 Mn(N(CN) 2 ) 2 (CH 3 OH) 2 ]·CH 3 OH (3) and [(NiL) 2 Mn 2 (N 3 ) 2 (μ 1,1 -N 3 ) 2 (CH 3 OH) 2 ] (4). Single crystal X-ray diffraction analyses show that complexes 2 and 3 have linear trinuclear structures where two tridentate O 3 donor (NiL) units are coordinated to the central octahedral Mn(ii) centre, whereas complex 4 has a centrosymmetric tetranuclear structure where two binuclear (NiL)Mn units are linked via two phenoxido and two μ 1,1 -N 3 bridges. Among the heterometallic complexes (2-4), only 4 is active towards the catalytic oxidation of 3,5-di-tert-butylcatechol to the corresponding quinone. The turnover number for the aerobic oxidation of 3,5-DTBC is 935 h -1 . ESI-mass spectra have been recorded to scrutinize the mechanistic pathway of this catalytic reaction. Variable temperature magnetic susceptibility measurements suggest that complexes 2-4 are antiferromagnetically coupled with coupling constants (J) of -4.84 and -5.23 cm -1 for complexes 2 and 3, respectively and J 1 = -2.20 cm -1 , J 2 = 1.13 cm -1 and J 3 = -1.12 cm -1 for complex 4. DFT calculations have been used to rationalize the magnetic super-exchange in complexes 2-4, by computing the theoretical coupling constants and analyzing the spin density plots.

Multiple sclerosis (MS) is a multifactorial, central nervous system, immune-mediated disease characterized by inflammation, demyelination, and neurodegeneration. Evidence suggests a steady rise in MS prevalence over the past five decades in the United States and around the world. Even with increased understanding of immunology, the specific etiological trigger of MS remains unknown. Evidence suggests that oxidative/nitroxidative stress is an important contributor to MS etiology, progression, and clinical symptoms. A multifaceted treatment approach aimed at counteracting oxidative/nitroxidative stress including MS disease-modifying medications, Mediterranean style diet, stress-relieving activities, smoking and alcohol cessation, exercise, and peer support programs is the best way to treat the disease.

Azanone (HNO/NO - ), also called nitroxyl, is a highly reactive compound whose biological role is still a matter of debate. A key issue that remains to be clarified regarding HNO and its biological activity is that of its endogenous formation. Given the overlap of the molecular targets and reactivity of nitric oxide (NO•) and HNO, its chemical biology was perceived to be similar to that of NO• as a biological signaling agent. However, despite their closely related reactivity, NO• and HNO's biochemical pathways are quite different. Moreover, the reduction of nitric oxide to azanone is possible but necessarily coupled to other reactions, which drive the reaction forward, overcoming the unfavorable thermodynamic barrier. The mechanism of this NO•/HNO interplay and its downstream effects in different contexts were studied recently, showing that more than fifteen moderate reducing agents react with NO• producing HNO. Particularly, it is known that the reaction between nitric oxide and hydrogen sulfide (H 2 S) produces HNO. However, this rate constant was not reported yet. In this work, firstly the NO•/H 2 S effective rate constant was measured as a function of the pH. Then, the implications of these chemical (non-enzymatic), biologically compatible, routes to endogenous HNO formation was discussed. There is no doubt that HNO could be (is?) a new endogenously produced messenger that mediates specific physiological responses, many of which were attributed yet to direct NO• effects., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Nitrate-rich food can increase nitric oxide production and improve vascular and brain functions. This study examines the feasibility of a randomised controlled trial (RCT) testing the effects of prolonged consumption of different doses of dietary nitrate (NO 3 - ) in the form of beetroot juice (BJ) in overweight and obese older participants. A single-blind, four-arm parallel pilot RCT was conducted in 62 overweight and obese (30.4 ± 4 kg/m 2 ) older participants (mean ± standard deviation (SD), 66 ± 4 years). Participants were randomized to: (1) high-NO 3 - (HN: 2 × 70 mL BJ/day) (2) medium-NO 3 - (MN: 70 mL BJ/day), (3) low-NO 3 - (LN: 70 mL BJ on alternate days) or (4) Placebo (PL: 70 mL of NO 3 - -depleted BJ on alternate days), for 13 weeks. Compliance was checked by a daily log of consumed BJ, NO 3 - intake, and by measuring NO 3 - and NO 2 - concentrations in plasma, saliva, and urine samples. Fifty participants completed the study. Self-reported compliance to the interventions was >90%. There were significant positive linear relationships between NO 3 - dose and the increase in plasma and urinary NO 3 - concentration (R 2 = 0.71, P < 0.001 and R 2 = 0.46 P < 0.001, respectively), but relationships between NO 3 - dose and changes in salivary NO 3 - and NO 2 - were non-linear (R 2 = 0.35, P = 0.002 and R 2 = 0.23, P = 0.007, respectively). The results confirm the feasibility of prolonged BJ supplementation in older overweight and obese adults., Competing Interests: The authors declare no conflict of interest.

Nitroaromatic antibiotics show activity against anaerobic bacteria and parasites, finding use in the treatment of Heliobacter pylori infections, tuberculosis, trichomoniasis, human African trypanosomiasis, Chagas disease and leishmaniasis. Despite this activity and a clear need for the development of new treatments for these conditions, the associated toxicity and lack of clear mechanisms of action have limited their therapeutic development. Nitroaromatic antibiotics require reductive bioactivation for activity and this reductive metabolism can convert the nitro group to nitric oxide (NO) or a related reactive nitrogen species (RNS). As nitric oxide plays important roles in the defensive immune response to bacterial infection through both signaling and redox-mediated pathways, defining controlled NO generation pathways from these antibiotics would allow the design of new therapeutics. This review focuses on the release of nitrogen oxide species from various nitroaromatic antibiotics to portend the increased ability for these compounds to positively impact infectious disease treatment.

We studied changes in LPO processes and antioxidant protection system in women with ovarian hyperandrogenism at various periods of reproductive age 18-45 years. An age-related decrease in total antioxidant activity and accumulation of secondary LPO products, ketodiens and coupled trienes, were found; these changes were accompanied by an increase in the level of superoxide dismutase and a decrease in the concentrations of reduced glutathione, α-tocopherol, and retinol. Women of early reproductive age (18-35 years) with hyperandrogenism were characterized by increased content of oxidized glutathione accompanied by a decrease in the level of α-tocopherol and retinol in comparison with the control group. In women of late reproductive age (35-45 years) with hyperandrogenism, increased superoxide dismutase activity in comparison with the control group was revealed.

We studied the effect of dinitrosyl-iron complexes with N-acetyl-L-cysteine as a thiol-containing ligand (DNIC-Acc) after transdermal administration to rats. Electron paramagnetic resonance spectroscopy with a lipophilic NO spin trap (a complex of iron and diethyldithiocarbamate ions) showed that DNIC-Acc administration significantly increased the total level of NO in the lung and liver tissues of the animal, which was accompanied by a slight decrease in the mean BP (<10%).

Genes coding for enzymes of the denitrification pathway appear randomly distributed among isolates of the ancestral genus Thermus , but only in few strains of the species Thermus thermophilus has the pathway been studied to a certain detail. Here, we review the enzymes involved in this pathway present in T. thermophilus NAR1, a strain extensively employed as a model for nitrate respiration, in the light of its full sequence recently assembled through a combination of PacBio and Illumina technologies in order to counteract the systematic errors introduced by the former technique. The genome of this strain is divided in four replicons, a chromosome of 2,021,843 bp, two megaplasmids of 370,865 and 77,135 bp and a small plasmid of 9799 pb. Nitrate respiration is encoded in the largest megaplasmid, pTTHNP4, within a region that includes operons for O 2 and nitrate sensory systems, a nitrate reductase, nitrate and nitrite transporters and a nitrate specific NADH dehydrogenase, in addition to multiple insertion sequences (IS), suggesting its mobility-prone nature. Despite nitrite is the final product of nitrate respiration in this strain, the megaplasmid encodes two putative nitrite reductases of the cd1 and Cu-containing types, apparently inactivated by IS. No nitric oxide reductase genes have been found within this region, although the NorR sensory gene, needed for its expression, is found near the inactive nitrite respiration system. These data clearly support that partial denitrification in this strain is the consequence of recent deletions and IS insertions in genes involved in nitrite respiration. Based on these data, the capability of this strain to transfer or acquire denitrification clusters by horizontal gene transfer is discussed.