Reactive oxygen species and NO-derived oxidizing, nitrosating and nitrating products mediate diverse cell signaling and pathologic processes in cardiovascular, pulmonary, and neurodegenerative diseases (1, 2). These reactive inflammatory mediators chemically modify carbohydrates, DNA bases, amino acids, and unsaturated fatty acids to form oxidized, nitrosated and nitrated derivatives. For example, accumulation of inflammatory-induced protein tyrosine nitration products represents a shift from the physiological signal-transducing actions of NO to an oxidative, nitrative, and potentially pathogenic pathway (1).Recently, it has been reported that nitration products of unsaturated fatty acids (nitroalkenes) are formed via NO-dependent oxidative reactions (3-5). These derivatives were initially viewed to be, like nitrotyrosine, a "footprint" of NO-dependent redox reactions (3, 6). More recently, we have observed that nitrolinoleate (LNO 2 ) 7 mediates pluripotent cell signaling actions, since it induces relaxation of phenylephrine-preconstricted rat aortic rings, inhibits thrombin-induced Ca 2ϩ elevations and aggregation of human platelets, and attenuates human neutrophil superoxide generation, degranulation, and integrin expression. These cell responses are mediated by both cGMP-and cAMP-dependent and -independent mechanisms (7-9).LNO 2 positional isomers, including 9-, 10-, 12-, and 13-nitro-9,12-cis-octadecadienoic acids, have been identified as free acids in human plasma and red blood cells and as esterified components of plasma lipoproteins and red blood cell membranes (10). In addition, plasma and red cell free and esterified nitrooleate (OA-NO 2 , isomers 9-and 10-nitro-9-cis-octadecenoic acid) was also identified in healthy human blood (11).Current knowledge indicates that enzymatically oxidized unsaturated fatty acid-derived products, such as prostaglandins, thromboxanes, leukotrienes, epoxyeicosatrienoic acids, hydroxyeicosatetraenoic acids, lipoxins, and resolvins serve as lipid mediators or autacoids. These signaling mediators act within a local microenvironment to orchestrate both physiological and pathological events, including platelet aggregation, constriction of vascular smooth muscle, neonatal development, wound healing, and resolution of inflammation (12, 13). In this context, endogenous nitrated fatty acid derivatives, such as * This work was supported by National Institutes of Health Grants HL068878, HL075397, and S06GM08248 (to Y. E. C.), HL70146 (to R. P. P.), and HL58115 and HL64937 (to B. A. F.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. □ S The on-line version of this article (available at http://www.jbc.org) contains supplemental Table S1, Fig. 1, and Movie 1. 1 Supported by American Diabetes Association Grant JFA 7-05-JF-12. 2 These authors contributed equally to this work. 3 Supported by the postdoctoral f...

Nitroalkene derivatives of linoleic acid (nitrolinoleic acid, LNO2) are formed via nitric oxide-dependent oxidative inflammatory reactions and are found at concentrations of Ϸ500 nM in the blood of healthy individuals. We report that LNO2 is a potent endogenous ligand for peroxisome proliferator-activated receptor ␥ (PPAR␥; Ki Ϸ133 nM) that acts within physiological concentration ranges. This nuclear hormone receptor (PPAR␥) regulates glucose homeostasis, lipid metabolism, and inflammation. PPAR␥ ligand activity is specific for LNO2 and not mediated by LNO2 decay products, NO donors, linoleic acid (LA), or oxidized LA. LNO2 is a significantly more robust PPAR␥ ligand than other reported endogenous PPAR␥ ligands, including lysophosphatidic acid (16:0 and 18:1), 15-deoxy-⌬ 12,14 -PGJ2, conjugated LA and azelaoyl-phosphocholine. LNO2 activation of PPAR␥ via CV-1 cell luciferase reporter gene expression analysis revealed a ligand activity that rivals or exceeds synthetic PPAR␥ agonists such as rosiglitazone and ciglitazone, is coactivated by 9 cis-retinoic acid and is inhibited by the PPAR␥ antagonist GW9662. LNO2 induces PPAR␥-dependent macrophage CD-36 expression, adipocyte differentiation, and glucose uptake also at a potency rivaling thiazolidinediones. These observations reveal that NO-mediated cell signaling reactions can be transduced by fatty acid nitration products and PPAR-dependent gene expression.fatty acid ͉ nitric oxide ͉ free radical ͉ adipocyte differentiation ͉ redox T he reaction of nitric oxide with tissue-free radical and oxidative intermediates yields secondary oxides of nitrogen that mediate oxidation, nitration, and nitrosation reactions (1, 2). Of present relevance, the reaction of NO and NO-derived species with oxidizing unsaturated fatty acids is kinetically rapid and exerts a multifaceted impact on cell redox and signaling reactions. NO readily outcompetes lipophilic antioxidants for the scavenging of lipid radicals, resulting in the inhibition of peroxyl radical-mediated chain propagation reactions (3). Both the catalytic activity and gene expression of eicosanoid biosynthetic enzymes are also regulated by NO, affirming a strong linkage between NO and fatty acid oxygenation product synthesis and signaling (4, 5). Consistent with this latter precept, fatty acid nitration products generated by NO-derived species inhibit multiple aspects of inflammatory cell function, indicating that nitrated fatty acids are both by-products and mediators of redox-signaling reactions (6-8).Recently, the structural characterization and quantitation of nitrolinoleic acid (LNO 2 ) in human red cells and plasma revealed this unsaturated fatty acid derivative to be the most abundant bioactive oxide of nitrogen in the vasculature. Net blood levels of Ϸ80 and 550 nM free and esterified LNO 2 , respectively, were measured in healthy humans (9). The observation that NO-dependent oxidative inflammatory reactions yield nitroalkene derivatives of unsaturated fatty acids displaying cGMP-independent cell signaling propert...

Abstract-Communication data rate and energy constraints are important factors which have to be considered when investigating distributed coordination of multi-agent networks. Although many proposed average-consensus protocols are available, a fundamental theoretic problem remains open, namely, how many bits of information are necessary for each pair of adjacent agents to exchange at each time step to ensure average consensus? In this paper, we consider average-consensus control of undirected networks of discrete-time first-order agents under communication constraints. Each agent has a real-valued state but can only exchange symbolic data with its neighbors. A distributed protocol is proposed based on dynamic encoding and decoding. It is proved that under the protocol designed, for a connected network, average consensus can be achieved with an exponential convergence rate based on merely one bit information exchange between each pair of adjacent agents at each time step. An explicit form of the asymptotic convergence rate is given. It is shown that as the number of agents increases, the asymptotic convergence rate is related to the scale of the network, the number of quantization levels and the ratio of the second smallest eigenvalue to the largest eigenvalue of the Laplacian of the communication graph. We also give a performance index to characterize the total communication energy to achieve average consensus and show that the minimization of the communication energy leads to a tradeoff between the convergence rate and the number of quantization levels.

Zinc-finger nuclease, transcription activator-like effector nuclease and CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated protein 9) are becoming major tools for genome editing. Importantly, knock-in in several non-rodent species has been finally achieved thanks to these customizable nucleases; yet the rates remain to be further improved. We hypothesize that inhibiting non-homologous end joining (NHEJ) or enhancing homology-directed repair (HDR) will improve the nuclease-mediated knock-in efficiency. Here we show that the in vitro application of an HDR enhancer, RS-1, increases the knock-in efficiency by two- to five-fold at different loci, whereas NHEJ inhibitor SCR7 has minimal effects. We then apply RS-1 for animal production and have achieved multifold improvement on the knock-in rates as well. Our work presents tools to nuclease-mediated knock-in animal production, and sheds light on improving gene-targeting efficiencies on pluripotent stem cells.