Nitric oxide from inflammatory origin impairs neural stem cell proliferation by inhibiting epidermal growth factor receptor signaling

Neuroinflammation is characterized by activation of microglial cells, followed by production of nitric oxide (NO), which may have different outcomes on neurogenesis, favoring or inhibiting this process. In the present study, we investigated how the inflammatory mediator NO can affect proliferation of neural stem cells (NSCs), and explored possible mechanisms underlying this effect. We investigated which mechanisms are involved in the regulation of NSC proliferation following treatment with an inflammatory stimulus (lipopolysaccharide plus IFN-gamma), using a culture system of subventricular zone (SVZ)-derived NSCs mixed with microglia cells obtained from wild-type mice (iNOS(+/+)) or from iNOS knockout mice (iNOS(-/-)). We found an impairment of NSC cell proliferation in iNOS(+/+) mixed cultures, which was not observed in iNOS(-/-) mixed cultures. Furthermore, the increased release of NO by activated iNOS(+/+) microglial cells decreased the activation of the ERK/MAPK signaling pathway, which was concomitant with an enhanced nitration of the EGF receptor. Preventing nitrogen reactive species formation with MnTBAP, a scavenger of peroxynitrite (ONOO-), or using the ONOO- degradation catalyst FeTMPyP cell proliferation and ERK signaling were restored to basal levels in iNOS(+/+) mixed cultures. Moreover, exposure to the NO donor NOC-18 (100 mu M), for 48 h, inhibited SVZ-derived NSC proliferation. Regarding the antiproliferative effect of NO, we found that NOC-18 caused the impairment of signaling through the ERK/MAPK pathway, which may be related to increased nitration of the EGF receptor in NSC. Using MnTBAP nitration was prevented, maintaining ERK signaling, rescuing NSC proliferation. We show that NO from inflammatory origin leads to a decreased function of the EGF receptor, which compromised proliferation of NSC. We also demonstrated that NO-mediated nitration of the EGF receptor caused a decrease in its phosphorylation, thus preventing regular proliferation signaling through the ERK/MAPK pathway.Foundation for Science and Technology, (FCT, Portugal); COMPETE; FEDER [PEst-C/SAU/LA0001/2013-2014, PEst-OE/EQB/LA0023/2013-2014, PTDC/SAU-NEU/102612/2008, PTDC/NEU-OSD/0473/2012]; FCT, Portugal [SERH/BPD/78901/2011, SERH/BD/38127/2007, SFRH/BD/77903/2011, SFRH/BD/79308/2011]info:eu-repo/semantics/publishedVersio

The Truth, the Whole Truth, and Nothing but the Truth: A Pragmatic Guide to Assessing Empirical Evaluations

An unsound claim can misdirect a field, encouraging the pursuit of unworthy ideas and the abandonment of promising ideas. An inadequate description of a claim can make it difficult to reason about the claim, for example to determine whether the claim is sound. Many practitioners will acknowledge the threat of un- sound claims or inadequate descriptions of claims to their field. We believe that this situation is exacerbated and even encouraged by the lack of a systematic approach to exploring, exposing, and addressing the source of unsound claims and poor exposition. This paper proposes a framework that identifies three sins of reasoning that lead to unsound claims and two sins of exposition that lead to poorly described claims. Sins of exposition obfuscate the objective of determining whether or not a claim is sound, while sins of reasoning lead directly to unsound claims. Our framework provides practitioners with a principled way of critiquing the integrity of their own work and the work of others. We hope that this will help individuals conduct better science and encourage a cultural shift in our research community to identify and promulgate sound claims

Path-sensitive data flow analysis simplified

Path-sensitive data flow analysis pairs classical data flow analysis with an analysis of feasibility of paths to improve precision. In this paper we propose a framework for path-sensitive backward data flow analysis that is enhanced with an abstraction of the predicate domain. The abstraction is based on a three-valued logic. It follows the strategy that path predicates are simplified if possible (without calling an external predicate solver) and every predicate that could not be reduced to a simple predicate is abstracted to the unknown value, for which the feasibility is undecided. The implementation of the framework scales well and delivers promising results