Molecular insights of p47phox phosphorylation dynamics in the regulation of NADPH oxidase activation and superoxide production

Phagocyte superoxide production by a multicomponent NADPH oxidase is important in host defense against microbial invasion. However inappropriate NADPH oxidase activation causes inflammation. Endothelial cells express NADPH oxidase and endothelial oxidative stress due to prolonged NADPH oxidase activation predisposes many diseases. Discovering the mechanism of NADPH oxidase activation is essential for developing novel treatment of these diseases. The p47phox is a key regulatory subunit of NADPH oxidase; however, due to the lack of full protein structural information, the mechanistic insight of p47phox phosphorylation in NADPH oxidase activation remains incomplete. Based on crystal structures of three functional domains, we generated a computational structural model of the full p47phox protein. Using a combination of in silico phosphorylation, molecular dynamics simulation and protein/protein docking, we discovered that the C-terminal tail of p47phox is critical for stabilizing its autoinhibited structure. Ser-379 phosphorylation disrupts H-bonds that link the C-terminal tail to the autoinhibitory region (AIR) and the tandem Src homology 3 (SH3) domains, allowing the AIR to undergo phosphorylation to expose the SH3 pocket for p22phox binding. These findings were confirmed by site-directed mutagenesis and gene transfection of p47phox_/_ coronary microvascular cells. Compared with wild-type p47phoxcDNAtransfected cells, the single mutation of S379A completely blocked p47phox membrane translocation, binding to p22phox and endothelial O2 . production in response to acute stimulation of PKC. p47phox C-terminal tail plays a key role in stabilizing intramolecular interactions at rest. Ser-379 phosphorylation is a molecular switch which initiates p47phox conformational changes and NADPH oxidase-dependent superoxide production by cells.

Temporally stable feature clusters for maritime object tracking in visible and thermal imagery

This paper describes a new approach to detect and track maritime objects in real time. The approach particularly addresses the highly dynamic maritime environment, panning cameras, target scale changes, and operates on both visible and thermal imagery. Object detection is based on agglomerative clustering of temporally stable features. Object extents are first determined based on persistence of detected features and their relative separation and motion attributes. An explicit cluster merging and splitting process handles object creation and separation. Stable object clus- ters are tracked frame-to-frame. The effectiveness of the approach is demonstrated on four challenging real-world public datasets.

Sahel decadal rainfall variability and the role of model horizontal resolution

Substantial low-frequency rainfall fluctuations occurred in the Sahel throughout the twentieth century, causing devastating drought. Modeling these low-frequency rainfall fluctuations has remained problematic for climate models for many years. Here we show using a combination of state-of-the-art rainfall observations and high-resolution global climate models that changes in organized heavy rainfall events carry most of the rainfall variability in the Sahel at multiannual to decadal time scales. Ability to produce intense, organized convection allows climate models to correctly simulate the magnitude of late-twentieth century rainfall change, underlining the importance of model resolution. Increasing model resolution allows a better coupling between large-scale circulation changes and regional rainfall processes over the Sahel. These results provide a strong basis for developing more reliable and skilful long-term predictions of rainfall (seasons to years) which could benefit many sectors in the region by allowing early adaptation to impending extremes.

An upgrade on the rabbit model of anthracycline-induced cardiomyopathy: shorter protocol, reduced mortality, and higher incidence of overt dilated cardiomyopathy

Current protocols of anthracycline-induced cardiomyopathy in rabbits present with high premature mortality and nephrotoxicity, thus rendering them unsuitable for studies requiring long-term functional evaluation of myocardial function (e.g., stem cell therapy). We compared two previously described protocols to an in-house developed protocol in three groups: Group DOX2 received doxorubicin 2 mg/kg/week (8 weeks); Group DAU3 received daunorubicin 3 mg/kg/week (10 weeks); and Group DAU4 received daunorubicin 4 mg/kg/week (6 weeks). A cohort of rabbits received saline (control). Results of blood tests, cardiac troponin I, echocardiography, and histopathology were analysed. Whilst DOX2 and DAU3 rabbits showed high premature mortality (50% and 33%, resp.), DAU4 rabbits showed 7.6% premature mortality. None of DOX2 rabbits developed overt dilated cardiomyopathy; 66% of DAU3 rabbits developed overt dilated cardiomyopathy and quickly progressed to severe congestive heart failure. Interestingly, 92% of DAU4 rabbits showed overt dilated cardiomyopathy and 67% developed congestive heart failure exhibiting stable disease. DOX2 and DAU3 rabbits showed alterations of renal function, with DAU3 also exhibiting hepatic function compromise. Thus, a shortened protocol of anthracycline-induced cardiomyopathy as in DAU4 group results in high incidence of overt dilated cardiomyopathy, which insidiously progressed to congestive heart failure, associated to reduced systemic compromise and very low premature mortality.

Interactions between wind-blown snow redistribution and melt ponds in a coupled ocean–sea ice model

Introducing a parameterization of the interactions between wind-driven snow depth changes and melt pond evolution allows us to improve large scale models. In this paper we have implemented an explicit melt pond scheme and, for the first time, a wind dependant snow redistribution model and new snow thermophysics into a coupled ocean–sea ice model. The comparison of long-term mean statistics of melt pond fractions against observations demonstrates realistic melt pond cover on average over Arctic sea ice, but a clear underestimation of the pond coverage on the multi-year ice (MYI) of the western Arctic Ocean. The latter shortcoming originates from the concealing effect of persistent snow on forming ponds, impeding their growth. Analyzing a second simulation with intensified snow drift enables the identification of two distinct modes of sensitivity in the melt pond formation process. First, the larger proportion of wind-transported snow that is lost in leads directly curtails the late spring snow volume on sea ice and facilitates the early development of melt ponds on MYI. In contrast, a combination of higher air temperatures and thinner snow prior to the onset of melting sometimes make the snow cover switch to a regime where it melts entirely and rapidly. In the latter situation, seemingly more frequent on first-year ice (FYI), a smaller snow volume directly relates to a reduced melt pond cover. Notwithstanding, changes in snow and water accumulation on seasonal sea ice is naturally limited, which lessens the impacts of wind-blown snow redistribution on FYI, as compared to those on MYI. At the basin scale, the overall increased melt pond cover results in decreased ice volume via the ice-albedo feedback in summer, which is experienced almost exclusively by MYI.

A review of drought in the Middle East and Southwest Asia

The Middle East and Southwest Asia comprise a region that is water-stressed, societally vulnerable, and prone to severe droughts. Large-scale climate variability, particularly La Niña, appears to play an important role in region-wide drought, including the two most severe of the last fifty years—1999-2001 and 2007-2008—with implications for drought forecasting. Important dynamical factors include orography, thermodynamic influence on vertical motion, storm track changes, and moisture transport. Vegetation in the region is strongly impacted by drought and may provide an important feedback mechanism. In future projections, drying of the eastern Mediterranean is a robust feature, as are temperature increases throughout the region, which will affect evaporation and the timing and intensity of snowmelt. Vegetation feedbacks may become more important in a warming climate. There are a wide range of outstanding issues for understanding, monitoring, and predicting drought in the region, including: dynamics of the regional storm track, the relative importance of the range of dynamical mechanisms related to drought, regional coherence of drought, the relationship between synoptic-scale mechanisms and drought, predictability of vegetation and crop yields, stability of remote influences, data uncertainty, and the role of temperature. Development of a regional framework for cooperative work and dissemination of information and existing forecasts would speed understanding and make better use of available information.

MEF2C-MYOCD and leiomodin1 suppression by miRNA-214 promotes smooth muscle cell phenotype switching in pulmonary arterial hypertension

Background Vascular hyperproliferative disorders are characterized by excessive smooth muscle cell (SMC) proliferation leading to vessel remodeling and occlusion. In pulmonary arterial hypertension (PAH), SMC phenotype switching from a terminally differentiated contractile to synthetic state is gaining traction as our understanding of the disease progression improves. While maintenance of SMC contractile phenotype is reportedly orchestrated by a MEF2C-myocardin (MYOCD) interplay, little is known regarding molecular control at this nexus. Moreover, the burgeoning interest in microRNAs (miRs) provides the basis for exploring their modulation of MEF2C-MYOCD signaling, and in turn, a pro-proliferative, synthetic SMC phenotype. We hypothesized that suppression of SMC contractile phenotype in pulmonary hypertension is mediated by miR-214 via repression of the MEF2C-MYOCD-leiomodin1 (LMOD1) signaling axis. Methods and Results In SMCs isolated from a PAH patient cohort and commercially obtained hPASMCs exposed to hypoxia, miR-214 expression was monitored by qRT-PCR. miR-214 was upregulated in PAH- vs. control subject hPASMCs as well as in commercially obtained hPASMCs exposed to hypoxia. These increases in miR-214 were paralleled by MEF2C, MYOCD and SMC contractile protein downregulation. Of these, LMOD1 and MEF2C were directly targeted by the miR. Mir-214 overexpression mimicked the PAH profile, downregulating MEF2C and LMOD1. AntagomiR-214 abrogated hypoxia-induced suppression of the contractile phenotype and its attendant proliferation. Anti-miR-214 also restored PAH-PASMCs to a contractile phenotype seen during vascular homeostasis. Conclusions Our findings illustrate a key role for miR-214 in modulation of MEF2C-MYOCD-LMOD1 signaling and suggest that an antagonist of miR-214 could mitigate SMC phenotype changes and proliferation in vascular hyperproliferative disorders including PAH.