Apurinic/apyrimidinic endonuclease-1/redox factor-1 (APE/Ref -1)

■ Abstract Cells in multicellular organisms are exposed to both endogenous oxidative stresses generated metabolically and to oxidative stresses that originate from neighboring cells and from other tissues. To protect themselves from oxidative stress, cells are equipped with reducing buffer systems (glutathione/GSH and thioredoxin/thioredoxin reductase) and have developed several enzymatic mechanisms against oxidants that include catalase, superoxide dismutase, and glutathione peroxidase. Other major extrinsic defenses (from the diet) include ascorbic acid, β-carotene and other carotenoids, and selenium. Recent evidence indicates that in addition to their antioxidant function, several of these redox species and systems are involved in regulation of biological processes, including cellular signaling, transcription factor activity, and apoptosis in normal and cancer cells. The survival and overall well-being of the cell is dependent upon the balance between the activity and the intracellular levels of these antioxidants as well as their interaction with various regulatory factors, including Ref-1, nuclear factor-κB, and activating protein-1.

A non-sinusoidal trajectory profile is proposed for the oscillating hydrofoil in the energy generators instead of conventional sinusoidal plunging/pitching motions to seek better energy extraction performance. The novel profile is achieved by combining a specially designed trapezoidal-like pitching motion with a sinusoidal plunging motion and investigated numerically on its output energy coefficient and total output efficiency. Through an adjustable parameter b, the pitching profile can be altered from a sinusoidal (b ¼ 1.0) to a square wave (b / N). In this work, a series of b ranging from 1.0 to 4.0 are investigated to examine the effect of combined motion trajectory on the energy extraction performance. The study encompasses the Strouhal numbers (St) from 0.05 to 0.5, nominal effective angle of attacks a0 of 10 and 20 and plunging amplitude h0/c of 0.5 and 1.0. Numerical results show that, for different b pitching motions, a larger a0 always results in a higher extraction power Cop and total efficiency hT. Compared with the sinusoidal motion (b ¼ 1), significant increment of Cop and hT can be observed for b > 1 over a certain range of St. The investigation also shows that there exists an optimal pitching profile which may increase the output power coefficient and total output efficiency as high as 63% and 50%, respectively, over a wide range of St. Detailed examination on the computed results reveal that, the energy extraction performance is determined by the relative ratio of the positive and negative contributions from the different combination of lift force, momentum and corresponding plunging velocity and pitching angular velocity, all of which are considerably affected by b

Key Points Question How has opioid use in pediatric patients changed over the past 10 years, and what factors are associated with differences in opioid use? Findings A cross-sectional study of 69 152 emergency department visits found that opioid prescribing rates decreased between 2006 to 2010 (8.23%) and 2011 to 2015 (6.30%). Region, race, age, and payment method were associated with differences in opioid prescribing. Meaning Opioid rates have decreased in recent years but inconsistencies in prescribing continue, suggesting the need for further research.

Nuclear factor kappa B (NFκB) is an essential regulator of gene transcription for hundreds of genes, including many critically involved in apoptosis. NFκB complexes containing cRel generally activate pro‐apoptotic genes, while those with RelA activate anti‐apoptotic genes. We have previously shown that NFκB binding by RelA is constitutively elevated in human metastatic melanoma cultures relative to normal melanocytes. Here we extended our investigation to immunohistochemical analysis of human tissue biopsies. We found that RelA expression is significantly elevated in melanocytes of human naevi and melanomas relative to normal skin, but expression of its inhibitor IκB‐α is significantly lower in metastatic melanomas than in intradermal naevi. Antibodies specific for the nuclear localization signal of RelA also showed significantly increased staining in metastatic melanoma biopsies. Notably, in melanomas and in naevi, we also found that RelA is phosphorylated at serine 529, and this activated form accumulates in the nuclei of melanomas. This suggests that increased expression and phosphorylation of RelA occurs at the stage of the benign naevus, but IκB‐α is able to sequester RelA in the cytoplasm and regulate RelA transcriptional transactivation. We also found that antibodies against cRel show a progressive increase in staining from naevi to melanoma. However, staining for IκB‐ɛ, which primarily inhibits the nuclear localization of cRel was also progressively increased, and cRel expression was predominantly cytoplasmic in melanomas. These results confirm that the altered expression of RelA found in metastatic melanoma cells in tissue culture is relevant to human tumors and offer new insights into the deregulation of NFκB signaling.

Aims: Our previous studies have shown that nitric oxide (NO) plays an important role in increasing the invasion and proliferation of human melanoma cells, suggesting that targeting NO signaling may facilitate therapy and prevention. Neuronal nitric oxide synthase (nNOS) is present in melanocytes, a cell type that originates from the neural crest. The aims of this study were to determine the role of nNOS in melanoma progression and the potential antitumor effects of novel synthesized nNOS inhibitors. Results: In vitro studies demonstrated abundant expression of nNOS in melanoma compared to melanocytes, which was inducible by ultraviolet radiation and was associated with increased NO generation. nNOS was also detected in melanoma biopsies that increased with disease stage. Knockdown of nNOS in melanoma cells diminished L-arginine-induced NO production; the metastatic capacity was also reduced as well as the levels of MMP-1, Bcl-2, JunD, and APE/Ref-1. Similar inhibition of NO and invasion potential was observed utilizing novel, highly selective nNOS inhibitors. In three-dimensional human skin reconstructs, the nNOS inhibitor cpd8 effectively reversed the melanoma overgrowth stimulated by NO stress. Innovation: Our work lays the foundation for development of clinical ''drug-like'' nNOS inhibitors as a new and promising strategy for the chemoprevention of early melanoma progression and the inhibition of secondary melanoma in high-risk individuals. Conclusion: Based on our observations, we propose that nNOS in melanoma results in constitutive overproduction of NO, which stimulates proliferation and increases invasion potential, leading to subsequent development of metastases.

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Selective inhibitors of neuronal nitric oxide synthase (nNOS) are regarded as valuable and powerful agents with therapeutic potential for the treatment of chronic neurodegenerative pathologies and human melanoma. Here, we describe a novel hybrid strategy that combines the pharmacokinetically promising thiophene-2-carboximidamide fragment and structural features of our previously reported potent and selective aminopyridine inhibitors. Two inhibitors, 13 and 14, show low nanomolar inhibitory potency (Ki = 5 nM for nNOS) and good isoform selectivities (nNOS over eNOS [440- and 540-fold, respectively] and over iNOS [260- and 340-fold, respectively]). The crystal structures of these nNOS–inhibitor complexes reveal a new hot spot that explains the selectivity of 14 and why converting the secondary to tertiary amine leads to enhanced selectivity. More importantly, these compounds are the first highly potent and selective nNOS inhibitory agents that exhibit excellent in vitro efficacy in melanoma cell lines.