Nitric oxide (NO) possesses antiinflammatory effects, which may be exerted via its ability to inhibit the transcription factor, NF-B. A commonly proposed mode of action for inhibition of NF-B by NO involves interference with NF-B binding to DNA. Because activation of inhibitory B kinase (IKK), the prerequisite enzyme complex necessary to induce NF-B, is subject to redox regulation, we assessed whether IKK could present a more proximal target for NO to inhibit NF-B activation. We demonstrate here that S-nitrosothiols (SNO) caused a dose-dependent inhibition of the enzymatic activity of IKK, in lung epithelial cells and in Jurkat T cells, which was associated with S-nitrosylation of the IKK complex. Using biotin derivatization of SNO, we revealed that IKK␤, the catalytic subunit required for NF-B activation, was a direct target for S-nitrosylation. A mutant version of IKK␤ containing a Cys-179-toAla mutation was refractory to inhibition by SNO or to increases in S-nitrosylation, in contrast to wild-type IKK␤, demonstrating that Cys-179 is the main target for attack by SNO. Importantly, inhibition of NO synthase activity in Jurkat T cells resulted in activation of IKK, in association with its denitrosylation. Moreover, NO synthase inhibition enhanced the ability of tumor necrosis factor ␣ to activate IKK, illustrating the importance of endogenous NO in regulating the extent of NF-B activation by cytokines. Collectively, our findings demonstrate that IKK␤ is an important target for the redox regulation of NF-B by endogenous or exogenous NO, providing an additional mechanism for its antiinflammatory properties.

Interleukin (IL)-1β is a cytokine critical to several inflammatory diseases in which pathogenic TH17 responses are implicated. Activation of the NLRP3 inflammasome by microbial and environmental stimuli can enable the caspase-1 dependent processing and secretion of IL-1β. The acute phase protein serum amyloid A (SAA) is highly induced during inflammatory responses, wherein it participates in systemic modulation of innate and adaptive immune responses. Elevated levels of IL-1β, SAA, and IL-17 are present in subjects with severe allergic asthma, yet the mechanistic relationship between these mediators has yet to be identified. Herein, we demonstrate that Saa3 is expressed in the lung of mice exposed to several mixed Th2/Th17-polarizing allergic sensitization regimens. SAA instillation into the lungs elicits robust TLR2-, MyD88-, and IL-1-dependent pulmonary neutrophilic inflammation. Furthermore, SAA drives production of IL-1α, IL-1β, IL-6, IL-23, and PGE2, causes dendritic cell maturation, and requires TLR2, MyD88, and the NLRP3 inflammasome for secretion of IL-1β by dendritic cells and macrophages. CD4+ T cells polyclonally stimulated in the presence of conditioned media from SAA-exposed dendritic cells produced IL-17 and the capacity of polyclonally-stimulated splenocytes to secrete IL-17 is dependent upon IL-1, TLR2, and the NLRP3 inflammasome. Additionally, in a model of allergic airway inflammation, administration of SAA to the lungs functions as an adjuvant to sensitize mice to inhaled ovalbumin, resulting in leukocyte influx after antigen challenge and a predominance of IL-17 production from restimulated splenocytes that is dependent upon IL-1 receptor signaling.

Pulmonary inflammation in asthma is orchestrated by the activity of NF-κB. NO and NO synthase (NOS) activity are important modulators of inflammation. The availability of the NOS substrate, l-arginine, is one of the mechanisms that controls the activity of NOS. Arginase also uses l-arginine as its substrate, and arginase-1 expression is highly induced in a murine model of asthma. Because we have previously described that arginase affects NOx content and interferes with the activation of NF-κB in lung epithelial cells, the goal of this study was to investigate the impact of arginase inhibition on the bioavailability of NO and the implications for NF-κB activation and inflammation in a mouse model of allergic airway disease. Administration of the arginase inhibitor BEC (S-(2-boronoethyl)-l-cysteine) decreased arginase activity and caused alterations in NO homeostasis, which were reflected by increases in S-nitrosylated and nitrated proteins in the lungs from inflamed mice. In contrast to our expectations, BEC enhanced perivascular and peribronchiolar lung inflammation, mucus metaplasia, NF-κB DNA binding, and mRNA expression of the NF-κB-driven chemokine genes CCL20 and KC, and lead to further increases in airways hyperresponsiveness. These results suggest that inhibition of arginase activity enhanced a variety of parameters relevant to allergic airways disease, possibly by altering NO homeostasis.

We investigated the antioxidant activity of phenylpropionic acids--caffeic (CAF), ferulic (FER), para-coumaric (COU) and cinnamic (CIN)--and phenolic acids and related compounds--gallic (GAL), methyl gallate (meGAL), vanillic (VAN) and gentisic (GEN)--using visible spectroscopy, inhibition of nitroblue tetrazolium (NBT) reduction, and electrochemical methods including cyclic voltammetry and potentiometry. In the spectroscopic assays, only CAF, GAL and meGAL were able to inhibit NBT reduction. The same compounds showed the lowest oxidation potentials (Epa) and the highest redox potentials deltaE) in the cyclic voltammetric and potentiometric studies, respectively. In addition, it was observed that the greater the number of hydroxyls linked to the aromatic ring, the greater was the antioxidant activity of the analysed compounds. The correlations of Spermann--used to compare the methods between themselves and the methods with the relationship structure-antioxidant activity--were r = -0.9762 for the cyclic voltammetric-potentiometric methods. r = 0.8333 for the inhibition of NBT reduction-potentiometric methods and r = -0.8095 for the inhibition of NBT reduction-cyclic voltammetric methods. The correlations for cyclic voltammetric, potentiometric and inhibition of NBT reduction methods-number of hydroxyls linked to the aromatic ring were r = -0.9636, 0.9636 and 0.9142, respectively. These findings indicate that the electrochemical methods together with spectroscopic studies are a good tool to evaluate the antioxidant activity of substances.

A wealth of recent studies points to the importance of airway epithelial cells in the orchestration of inflammatory responses in the allergic inflamed lung. Studies also point to a role of oxidative stress in the pathophysiology of chronic inflammatory diseases. This article provides a perspective on the significance of airway epithelial cells in allergic inflammation, and reviews the relevance of the transcription factor, nuclear factor kappaB, herein. We also provide the reader with a perspective on the role that oxidants can play in lung homeostasis, and address the concept of "redox biology." In addition, we review recent evidence that highlights potential inhibitory roles of oxidants on nuclear factor kappaB activation and inflammation, and discuss recent assays that have become available to probe the functional roles of oxidants in lung biology.

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O comportamento eletroquímico do ácido cafeico (H 3 CAF) em meio aquoso foi estudado na faixa de pH 2,0-8,5 aplicando-se as técnicas voltametria cíclica, eletrólise com potencial controlado e espectroscopia UV-vis. A eletro-oxidação envolve a transferência reversível de dois elétrons e de dois prótons em soluções de pH até 5,5, de acordo com o mecanismo uma etapa-dois elétrons. Em soluções de pH superiores a 5,5 a eletro-oxidação do H 3 CAF segue um mecanismo EC i . O principal produto desta oxidação é a o-quinona correspondente (o-HCAF), a qual decompõe-se rapidamente em soluções de pH superior a 7,4, obedecendo a uma cinética de primeira ordem. Na faixa de pH 2,0-8,5, o potencial formal (E 0' ) varia linearmente com o pH, gerando uma reta com coeficiente angular de -60,83 mV/pH. Em paralelo, a corrente de pico anódica (i pa ) diminui de modo não-linear. Os processos são controlados por difusão em toda a faixa de pH estudada.The electrochemical behavior of caffeic acid (H 3 CAF) in aqueous solutions with pH 2.0 to 8.5 was studied by cyclic voltammetry, controlled potential electrolysis and UV-vis spectroscopy. The electrooxidation of H 3 CAF involves a reversible transfer of two electrons and two protons in solutions of pH up to 5.5, in agreement with the one step-two electron mechanism. In solutions of pH higher than 5.5, the process of electro-oxidation of H 3 CAF follows an EC i mechanism. The main oxidation product is the corresponding o-quinone (o-HCAF), which is decomposed quickly at pH higher than 7.4 obeying a first order kinetics. In the pH range investigated, the formal potential (E 0' ) varies linearly with pH, generating a straight line with an angular coefficient of -60.83 mV/pH. In parallel, the anodic peak current (i pa ) decreases in a nonlinear mode. The processes are controlled by diffusion over the whole pH range studied.

While a number of recent studies have indicated that CNCs are promising nanomaterials for biomedical applications, there is a substantial amount of work that still remains to be done before realizing the full therapeutic potential of CNCs. Major effort should be focused on detailed in vitro and in vivo studies of modified CNCs constructs in order to better understand the integration of CNCs in the biological systems.