Assessment of the Activation State of Rho Family GTP-Binding Proteins in Breast Cancer Cells and Specimens

DTIC Science & Technology

2004-08-01

AF038388 Vav3 GEF for Rho and Rac Proto-oncogene product NM020505 hPEM -2 GEF for Cdc42 Predominantly expressed in brain AB007884 GEF-H1 GEF for Rac and Rho...fence tions, and desmosomes play a fundamental role in maintain- function). A functional tight junction is crucial to maintain ing the polarized phenotype

Activated RhoA Is a Positive Feedback Regulator of the Lbc Family of Rho Guanine Nucleotide Exchange Factor Proteins*

PubMed Central

Medina, Frank; Carter, Angela M.; Dada, Olugbenga; Gutowski, Stephen; Hadas, Jana; Chen, Zhe; Sternweis, Paul C.

2013-01-01

The monomeric Rho GTPases are essential for cellular regulation including cell architecture and movement. A direct mechanism for hormonal regulation of the RhoA-type GTPases is their modulation by the G12 and G13 proteins via RH (RGS homology) containing RhoGEFs. In addition to the interaction of the G protein α subunits with the RH domain, activated RhoA also binds to the pleckstrin homology (PH) domain of PDZRhoGEF. The latter interaction is now extended to all seven members of the homologous Lbc family of RhoGEFs which includes the RH-RhoGEFs. This is evinced by direct measurements of binding or through effects on selected signaling pathways in cells. Overexpression of these PH domains alone can block RhoA-dependent signaling in cells to various extents. Whereas activated RhoA does not modulate the intrinsic activity of the RhoGEFs, activated RhoA associated with phospholipid vesicles can facilitate increased activity of soluble RhoGEFs on vesicle-delimited substrate (RhoA-GDP). This demonstrates feasibility of the hypothesis that binding of activated RhoA to the PH domains acts as a positive feedback mechanism. This is supported by cellular studies in which mutation of this binding site on PH strongly attenuates the stimulation of RhoA observed by overexpression of five of the RhoGEF DH-PH domains. This mutation is even more dramatic in the context of full-length p115RhoGEF. The utilization of this mechanism by multiple RhoGEFs suggests that this regulatory paradigm may be a common feature in the broader family of RhoGEFs. PMID:23493395

A High-Throughput Assay for Rho Guanine Nucleotide Exchange Factors Based on the Transcreener GDP Assay.

PubMed

Reichman, Melvin; Schabdach, Amanda; Kumar, Meera; Zielinski, Tom; Donover, Preston S; Laury-Kleintop, Lisa D; Lowery, Robert G

2015-12-01

Ras homologous (Rho) family GTPases act as molecular switches controlling cell growth, movement, and gene expression by cycling between inactive guanosine diphosphate (GDP)- and active guanosine triphosphate (GTP)-bound conformations. Guanine nucleotide exchange factors (GEFs) positively regulate Rho GTPases by accelerating GDP dissociation to allow formation of the active, GTP-bound complex. Rho proteins are directly involved in cancer pathways, especially cell migration and invasion, and inhibiting GEFs holds potential as a therapeutic strategy to diminish Rho-dependent oncogenesis. Methods for measuring GEF activity suitable for high-throughput screening (HTS) are limited. We developed a simple, generic biochemical assay method for measuring GEF activity based on the fact that GDP dissociation is generally the rate-limiting step in the Rho GTPase catalytic cycle, and thus addition of a GEF causes an increase in steady-state GTPase activity. We used the Transcreener GDP Assay, which relies on selective immunodetection of GDP, to measure the GEF-dependent stimulation of steady-state GTP hydrolysis by small GTPases using Dbs (Dbl's big sister) as a GEF for Cdc42, RhoA, and RhoB. The assay is well suited for HTS, with a homogenous format and far red fluorescence polarization (FP) readout, and it should be broadly applicable to diverse Rho GEF/GTPase pairs. © 2015 Society for Laboratory Automation and Screening.

Stimulation of Cortical Myosin Phosphorylation by p114RhoGEF Drives Cell Migration and Tumor Cell Invasion

PubMed Central

Zihni, Ceniz; Harris, Andrew R.; Bailly, Maryse; Charras, Guillaume T.; Balda, Maria S.; Matter, Karl

2012-01-01

Actinomyosin activity is an important driver of cell locomotion and has been shown to promote collective cell migration of epithelial sheets as well as single cell migration and tumor cell invasion. However, the molecular mechanisms underlying activation of cortical myosin to stimulate single cell movement, and the relationship between the mechanisms that drive single cell locomotion and those that mediate collective cell migration of epithelial sheets are incompletely understood. Here, we demonstrate that p114RhoGEF, an activator of RhoA that associates with non-muscle myosin IIA, regulates collective cell migration of epithelial sheets and tumor cell invasion. Depletion of p114RhoGEF resulted in specific spatial inhibition of myosin activation at cell-cell contacts in migrating epithelial sheets and the cortex of migrating single cells, but only affected double and not single phosphorylation of myosin light chain. In agreement, overall elasticity and contractility of the cells, processes that rely on persistent and more constant forces, were not affected, suggesting that p114RhoGEF mediates process-specific myosin activation. Locomotion was p114RhoGEF-dependent on Matrigel, which favors more roundish cells and amoeboid-like actinomyosin-driven movement, but not on fibronectin, which stimulates flatter cells and lamellipodia-driven, mesenchymal-like migration. Accordingly, depletion of p114RhoGEF led to reduced RhoA, but increased Rac activity. Invasion of 3D matrices was p114RhoGEF-dependent under conditions that do not require metalloproteinase activity, supporting a role of p114RhoGEF in myosin-dependent, amoeboid-like locomotion. Our data demonstrate that p114RhoGEF drives cortical myosin activation by stimulating myosin light chain double phosphorylation and, thereby, collective cell migration of epithelial sheets and amoeboid-like motility of tumor cells. PMID:23185572

Regulation of Ras Exchange Factors and Cellular Localization of Ras Activation by Lipid Messengers in T Cells

PubMed Central

Jun, Jesse E.; Rubio, Ignacio; Roose, Jeroen P.

2013-01-01

The Ras-MAPK signaling pathway is highly conserved throughout evolution and is activated downstream of a wide range of receptor stimuli. Ras guanine nucleotide exchange factors (RasGEFs) catalyze GTP loading of Ras and play a pivotal role in regulating receptor-ligand induced Ras activity. In T cells, three families of functionally important RasGEFs are expressed: RasGRF, RasGRP, and Son of Sevenless (SOS)-family GEFs. Early on it was recognized that Ras activation is critical for T cell development and that the RasGEFs play an important role herein. More recent work has revealed that nuances in Ras activation appear to significantly impact T cell development and selection. These nuances include distinct biochemical patterns of analog versus digital Ras activation, differences in cellular localization of Ras activation, and intricate interplays between the RasGEFs during distinct T cell developmental stages as revealed by various new mouse models. In many instances, the exact nature of these nuances in Ras activation or how these may result from fine-tuning of the RasGEFs is not understood. One large group of biomolecules critically involved in the control of RasGEFs functions are lipid second messengers. Multiple, yet distinct lipid products are generated following T cell receptor (TCR) stimulation and bind to different domains in the RasGRP and SOS RasGEFs to facilitate the activation of the membrane-anchored Ras GTPases. In this review we highlight how different lipid-based elements are generated by various enzymes downstream of the TCR and other receptors and how these dynamic and interrelated lipid products may fine-tune Ras activation by RasGEFs in developing T cells. PMID:24027568

Short-term Drought Prediction in India.

NASA Astrophysics Data System (ADS)

Shah, R.; Mishra, V.

2014-12-01

Medium range soil moisture drought forecast helps in decision making in the field of agriculture and water resources management. Part of skills in medium range drought forecast comes from precipitation. Proper evaluation and correction of precipitation forecast may improve drought predictions. Here, we evaluate skills of ensemble mean precipitation forecast from Global Ensemble Forecast System (GEFS) for medium range drought predictions over India. Climatological mean (CLIM) of historic data (OBS) are used as reference forecast to evaluate GEFS precipitation forecast. Analysis was conducted based on forecast initiated on 1st and 15th dates of each month for lead up to 7-days. Correlation and RMSE were used to estimate skill scores of accumulated GEFS precipitation forecast from lead 1 to 7-days. Volumetric indices based on the 2X2 contingency table were used to check missed and falsely predicted historic volume of daily precipitation from GEFS in different regions and at different thresholds. GEFS showed improvement in correlation of 0.44 over CLIM during the monsoon season and 0.55 during the winter season. Lower RMSE was showed by GEFS than CLIM. Ratio of RMSE in GEFS and CLIM comes out as 0.82 and 0.4 (perfect skill is at zero) during the monsoon and winter season, respectively. We finally used corrected GEFS forecast to derive the Variable Infiltration Capacity (VIC) model, which was used to develop short-term forecast of hydrologic and agricultural (soil moisture) droughts in India.

The Putative Exchange Factor Gef3p Interacts with Rho3p GTPase and the Septin Ring during Cytokinesis in Fission Yeast*

PubMed Central

Muñoz, Sofía; Manjón, Elvira; Sánchez, Yolanda

2014-01-01

The small GTP-binding proteins of the Rho family and its regulatory proteins play a central role in cytokinetic actomyosin ring assembly and cytokinesis. Here we show that the fission yeast guanine nucleotide exchange factor Gef3p interacts with Rho3p at the division site. Gef3p contains a putative DH homology domain and a BAR/IMD-like domain. The protein localized to the division site late in mitosis, where it formed a ring that did not constrict with actomyosin ring (cytokinetic actomyosin ring) invagination; instead, it split into a double ring that resembled the septin ring. Gef3p co-localized with septins and Mid2p and required septins and Mid2p for its localization. Gef3p interacts physically with the GTP-bound form of Rho3p. Although Gef3p is not essential for cell separation, the simultaneous disruption of gef3+ and Rho3p-interacting proteins, such as Sec8p, an exocyst component, Apm1p, a subunit of the clathrin adaptor complex or For3p, an actin-polymerizing protein, yielded cells with strong defects in septation and polarity respectively. Our results suggest that interactions between septins and Rho-GEFs provide a new targeting mechanism for GTPases in cytokinesis, in this case probably contributing to Rho3p function in vesicle tethering and vesicle trafficking in the later steps of cell separation. PMID:24947517

[Subgroup Analysis of the Non-interventional REASON Study: PFS and OS According to Age, Smoking History, Gender, and Histology in NSCLC Patients Treated with Gefitinib or Chemotherapy].

PubMed

Schuette, W; Eberhardt, W E E; Waller, C; Schirmacher, P; Dietel, M; Zirrgiebel, U; Radke, S; Thomas, M

2016-09-01

Assessment of several clinical factors on progression-free (PFS) and overall survival (OS) in NSCLC patients (pts.) (stage IV) with mutated epidermal growth factor receptor (EGFRm+) treated with gefitinib (gef) or with chemotherapy (CT) under real-world conditions. 285 EGFRm+ pts. of the non-interventional REASON study treated with gef (n = 206) or CT (n = 79) as first-line therapy or with gef (n = 213) or CT (n = 61) in any line throughout the course of therapy were analyzed according to age, gender, smoking history and histology. Compared with CT, patients treated with gef showed prolongation of PFS and OS in all subgroups. PFS was significantly increased in women and non-smokers. OS was significantly increased in women, non-smokers, (ex)-smokers, patients with adenocarcinoma and elderly patients when treated with gef compared to CT. Female gender turned out to be an independent positive predictive factor for OS in patients treated with gef (HRmale: 1.74, p = 0.0009). A clinical benefit of gef was shown for all analyzed clinical subgroups of EGFRm+ pts. This was confirmed for the female gender in a multivariate analysis. © Georg Thieme Verlag KG Stuttgart · New York.

Science of Integrated Approaches to Natural Resources Management

NASA Astrophysics Data System (ADS)

Tengberg, Anna; Valencia, Sandra

2017-04-01

To meet multiple environmental objectives, integrated programming is becoming increasingly important for the Global Environmental Facility (GEF), the financial mechanism of the multilateral environmental agreements, including the United Nations Convention to Combat Desertification (UNCCD). Integration of multiple environmental, social and economic objectives also contributes to the achievement of the Sustainable Development Goals (SDGs) in a timely and cost-effective way. However, integration is often not well defined. This paper therefore focuses on identifying key aspects of integration and assessing their implementation in natural resources management (NRM) projects. To that end, we draw on systems thinking literature, and carry out an analysis of a random sample of GEF integrated projects and in-depth case studies demonstrating lessons learned and good practices in addressing land degradation and other NRM challenges. We identify numerous challenges and opportunities of integrated approaches that need to be addressed in order to maximise the catalytic impact of the GEF during problem diagnosis, project design, implementation and governance. We highlight the need for projects to identify clearer system boundaries and main feedback mechanisms within those boundaries, in order to effectively address drivers of environmental change. We propose a theory of change for Integrated Natural Resources Management (INRM) projects, where short-term environmental and socio-economic benefits will first accrue at the local level. Implementation of improved INRM technologies and practices at the local level can be extended through spatial planning, strengthening of innovation systems, and financing and incentive mechanisms at the watershed and/or landscape/seascape level to sustain and enhance ecosystem services at larger scales and longer time spans. We conclude that the evolving scientific understanding of factors influencing social, technical and institutional innovations and transitions towards sustainable management of natural resources should be harnessed and integrated into GEF's influencing models and theory of change, and be coupled with updated approaches for learning, adaptive management and scaling up.

Release of GTP Exchange Factor Mediated Down-Regulation of Abscisic Acid Signal Transduction through ABA-Induced Rapid Degradation of RopGEFs

PubMed Central

Waadt, Rainer; Schroeder, Julian I.

2016-01-01

The phytohormone abscisic acid (ABA) is critical to plant development and stress responses. Abiotic stress triggers an ABA signal transduction cascade, which is comprised of the core components PYL/RCAR ABA receptors, PP2C-type protein phosphatases, and protein kinases. Small GTPases of the ROP/RAC family act as negative regulators of ABA signal transduction. However, the mechanisms by which ABA controls the behavior of ROP/RACs have remained unclear. Here, we show that an Arabidopsis guanine nucleotide exchange factor protein RopGEF1 is rapidly sequestered to intracellular particles in response to ABA. GFP-RopGEF1 is sequestered via the endosome-prevacuolar compartment pathway and is degraded. RopGEF1 directly interacts with several clade A PP2C protein phosphatases, including ABI1. Interestingly, RopGEF1 undergoes constitutive degradation in pp2c quadruple abi1/abi2/hab1/pp2ca mutant plants, revealing that active PP2C protein phosphatases protect and stabilize RopGEF1 from ABA-mediated degradation. Interestingly, ABA-mediated degradation of RopGEF1 also plays an important role in ABA-mediated inhibition of lateral root growth. The presented findings point to a PP2C-RopGEF-ROP/RAC control loop model that is proposed to aid in shutting off ABA signal transduction, to counteract leaky ABA signal transduction caused by “monomeric” PYL/RCAR ABA receptors in the absence of stress, and facilitate signaling in response to ABA. PMID:27192441

Signaling efficiency of Gαq through its effectors p63RhoGEF and GEFT depends on their subcellular location.

PubMed

Goedhart, Joachim; van Unen, Jakobus; Adjobo-Hermans, Merel J W; Gadella, Theodorus W J

2013-01-01

The p63RhoGEF and GEFT proteins are encoded by the same gene and both members of the Dbl family of guanine nucleotide exchange factors. These proteins can be activated by the heterotrimeric G-protein subunit Gαq. We show that p63RhoGEF is located at the plasma membrane, whereas GEFT is confined to the cytoplasm. Live-cell imaging studies yielded quantitative information on diffusion coefficients, association rates and encounter times of GEFT and p63RhoGEF. Calcium signaling was examined as a measure of the signal transmission, revealing more efficient signaling through the membrane-associated p63RhoGEF. A rapamycin dependent recruitment system was used to dynamically alter the subcellular location and concentration of GEFT, showing efficient signaling through GEFT only upon membrane recruitment. Together, our results show efficient signal transmission through membrane located effectors, and highlight a role for increased concentration rather than increased encounter times due to membrane localization in the Gαq mediated pathways to p63RhoGEF and PLCβ.

Assessment of the Activation of Rho Family GTP-Binding Proteins in Breast Cancer Cells and Specimens

DTIC Science & Technology

2001-08-01

lymphopenia Vav2 GEF for Rho, Rac, and Cdc42 proto-oncogene product; NM009500 Vav3 GEF for Rho and Rac proto-oncogene product; NM020505 hPEM -2 GEF for...junctions, and desmosomes play a fundamental role in maintaining the polarized phenotype and vectorial transport functions of epithelial cells. The tight

RhoA GTPase inhibition organizes contraction during epithelial morphogenesis

PubMed Central

Mason, Frank M.; Xie, Shicong; Vasquez, Claudia G.; Tworoger, Michael

2016-01-01

During morphogenesis, contraction of the actomyosin cytoskeleton within individual cells drives cell shape changes that fold tissues. Coordination of cytoskeletal contractility is mediated by regulating RhoA GTPase activity. Guanine nucleotide exchange factors (GEFs) activate and GTPase-activating proteins (GAPs) inhibit RhoA activity. Most studies of tissue folding, including apical constriction, have focused on how RhoA is activated by GEFs to promote cell contractility, with little investigation as to how GAPs may be important. Here, we identify a critical role for a RhoA GAP, Cumberland GAP (C-GAP), which coordinates with a RhoA GEF, RhoGEF2, to organize spatiotemporal contractility during Drosophila melanogaster apical constriction. C-GAP spatially restricts RhoA pathway activity to a central position in the apical cortex. RhoGEF2 pulses precede myosin, and C-GAP is required for pulsation, suggesting that contractile pulses result from RhoA activity cycling. Finally, C-GAP expression level influences the transition from reversible to irreversible cell shape change, which defines the onset of tissue shape change. Our data demonstrate that RhoA activity cycling and modulating the ratio of RhoGEF2 to C-GAP are required for tissue folding. PMID:27551058

The Amino-Terminus of Vav1 Regulates TCR-Induced SLP-76 Microclusters via the Calponin Homology Domain and Non-Catalytic Surfaces within the GEF Module

PubMed Central

Sylvain, Nicholas R.; Nguyen, Ken; Bunnell, Stephen C.

2013-01-01

The guanine nucleotide exchange factor (GEF) Vav1 synergizes with the adapter SLP-76 to support T cell development and activation. Here, we demonstrate that Vav1 controls the stability and movement T cell receptor-induced SLP-76 microclusters. The SH2 domain enables the recruitment of Vav1 into SLP-76 microclusters, whereas the SH3 domains of Vav1 cooperate to enhance microcluster stability and function. Although the amino-terminus of Vav1 is essential for downstream signaling, it possesses novel scaffolding functions that are unaffected by the inactivation of the Vav1 GEF or by the constitutive GEF activation that accompanies the mutation of the regulatory tyrosine residues 142, 160, and 174. In contrast, GEF-inactivating point mutations predicted to perturb the structural integrity of the Vav1 GEF abolish these scaffolding functions. Paradoxically, the excision of catalytic Dbl-homology (DH) / pleckstrin homology (PH) cassette produces a relatively mild scaffolding defect, indicating that the L213A and L278Q point mutations antagonize scaffolding functions mediated by adjacent domains. A deletion mutant lacking the CH domain potently inhibits calcium responses, but also exhibits mild scaffolding defects. We conclude multiple GEF-independent scaffolding functions contained within the amino-terminus of Vav1 contribute to T cell activation by acting synergistically to increase the stability and functionality of SLP-76 microclusters. PMID:21386095

The Impact of STTP on the GEFS Forecast of Week-2 and Beyond in the Presence of Stochastic Physics

NASA Astrophysics Data System (ADS)

Hou, D.

2015-12-01

The Stochastic Total Tendency Perturbation (STTP) scheme was designed to represent the model related uncertainties not considered in the numerical model itself and the physics based stochastic schemes. It has been applied in NCEP's Global Ensemble Forecast System (GEFS) since 2010, showing significant positive impacts on the forecast with improved spread-error ratio and probabilistic forecast skills. The scheme is robust and it went well with the resolution increases and model improvements in 2012 and 2015 with minimum changes. Recently, a set of stochastic physics schemes are coded in the Global Forecast System model and tested in the GEFS package. With these schemes turned on and STTP off, the forecast performance is comparable or even superior to the operational GEFS, in which STTP is the only contributor to the model related uncertainties. This is true especially in week one. However, over the second week and beyond, both the experimental and the operational GEFS has insufficient spread, especially over the warmer seasons. This is a major challenge when the GEFS is extended to sub-seasonal (week 4-6) time scales. The impact of STTP on the GEFS forecast in the presence of stochastic physics is investigated by turning both the stochastic physics schemes and STTP on and carefully tuning their amplitudes. Analysis will be focused on the forecast of extended range, especially week 2. Its impacts on week 3-4 will also be addressed.

Role of Longwave Cloud-Radiation Feedback in the Simulation of the Madden-Julian Oscillation

NASA Technical Reports Server (NTRS)

Kim, Daehyun; Ahn, Min-Seop; Kang, In-Sik; Del Genio, Anthony D.

2015-01-01

The role of the cloud-radiation interaction in the simulation of the Madden-Julian oscillation (MJO) is investigated. A special focus is on the enhancement of column-integrated diabatic heating due to the greenhouse effects of clouds and moisture in the region of anomalous convection. The degree of this enhancement, the greenhouse enhancement factor (GEF), is measured at different precipitation anomaly regimes as the negative ratio of anomalous outgoing longwave radiation to anomalous precipitation. Observations show that the GEF varies significantly with precipitation anomaly and with the MJO cycle. The greenhouse enhancement is greater in weak precipitation anomaly regimes and its effectiveness decreases monotonically with increasing precipitation anomaly. The GEF also amplifies locally when convection is strengthened in association with the MJO, especially in the weak precipitation anomaly regime (less than 5 mm day(exp -1)). A robust statistical relationship is found among CMIP5 climate model simulations between the GEF and the MJO simulation fidelity. Models that simulate a stronger MJO also simulate a greater GEF, especially in the weak precipitation anomaly regime (less than 5 mm day(exp -1)). Models with a greater GEF in the strong precipitation anomaly regime (greater than 30 mm day(-1)) represent a slightly slower MJO propagation speed. Many models that lack the MJO underestimate the GEF in general and in particular in the weak precipitation anomaly regime. The results herein highlight that the cloud-radiation interaction is a crucial process for climate models to correctly represent the MJO.

Activation of p115-RhoGEF Requires Direct Association of G[alpha subscript 13] and the Dbl Homology Domain

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chen, Zhe; Guo, Liang; Hadas, Jana

2012-09-05

RGS-containing RhoGEFs (RGS-RhoGEFs) represent a direct link between the G{sub 12} class of heterotrimeric G proteins and the monomeric GTPases. In addition to the canonical Dbl homology (DH) and pleckstrin homology domains that carry out the guanine nucleotide exchange factor (GEF) activity toward RhoA, these RhoGEFs also possess RGS homology (RH) domains that interact with activated {alpha} subunits of G{sub 12} and G{sub 13}. Although the GEF activity of p115-RhoGEF (p115), an RGS-RhoGEF, can be stimulated by G{alpha}{sub 13}, the exact mechanism of the stimulation has remained unclear. Using combined studies with small angle x-ray scattering, biochemistry, and mutagenesis, wemore » identify an additional binding site for activated G{alpha}{sub 13} in the DH domain of p115. Small angle x-ray scattering reveals that the helical domain of G{alpha}{sub 13} docks onto the DH domain, opposite to the surface of DH that binds RhoA. Mutation of a single tryptophan residue in the {alpha}3b helix of DH reduces binding to activated G{alpha}{sub 13} and ablates the stimulation of p115 by G{alpha}{sub 13}. Complementary mutations at the predicted DH-binding site in the {alpha}B-{alpha}C loop of the helical domain of G{alpha}{sub 13} also affect stimulation of p115 by G{alpha}{sub 13}. Although the GAP activity of p115 is not required for stimulation by G{alpha}{sub 13}, two hydrophobic motifs in RH outside of the consensus RGS box are critical for this process. Therefore, the binding of G{alpha}{sub 13} to the RH domain facilitates direct association of G{alpha}{sub 13} to the DH domain to regulate its exchange activity. This study provides new insight into the mechanism of regulation of the RGS-RhoGEF and broadens our understanding of G protein signaling.« less

Function of the nucleotide exchange activity of vav1 in T cell development and activation.

PubMed

Saveliev, Alexander; Vanes, Lesley; Ksionda, Olga; Rapley, Jonathan; Smerdon, Stephen J; Rittinger, Katrin; Tybulewicz, Victor L J

2009-12-15

The guanine nucleotide exchange factor (GEF) Vav1 is essential for transducing T cell antigen receptor (TCR) signals and therefore plays a critical role in the development and activation of T cells. It has been presumed that the GEF activity of Vav1 is important for its function; however, there has been no direct demonstration of this. Here, we generated mice expressing enzymatically inactive, but normally folded, Vav1 protein. Analysis of these mice showed that the GEF activity of Vav1 was necessary for the selection of thymocytes and for the optimal activation of T cells, including signal transduction to Rac1, Akt, and integrins. In contrast, the GEF activity of Vav1 was not required for TCR-induced calcium flux, activation of extracellular signal-regulated kinase and protein kinase D1, and cell polarization. Thus, in T cells, the GEF activity of Vav1 is essential for some, but not all, of its functions.

Function of the Nucleotide Exchange Activity of Vav1 in T cell Development and Activation*

PubMed Central

Saveliev, Alexander; Vanes, Lesley; Ksionda, Olga; Rapley, Jonathan; Smerdon, Stephen J.; Rittinger, Katrin; Tybulewicz, Victor L. J.

2012-01-01

The guanine nucleotide exchange factor (GEF) Vav1 is essential for transducing T cell antigen receptor (TCR) signals and therefore plays a critical role in the development and activation of T cells. It has been presumed that the GEF activity of Vav1 is important for its function; however, there has been no direct demonstration of this. Here, we generated mice expressing enzymatically inactive, but normally folded, Vav1 protein. Analysis of these mice showed that the GEF activity of Vav1 was necessary for the selection of thymocytes and for the optimal activation of T cells, including signal transduction to Rac1, Akt, and integrins. In contrast, the GEF activity of Vav1 was not required for TCR-induced calcium flux, activation of extracellular signal–regulated kinase (ERK) and protein kinase D1 (PKD1), and cell polarization. Thus, in T cells, the GEF activity of Vav1 is essential for some, but not all, of its functions. PMID:20009105

Son of sevenless directly links the Robo receptor to rac activation to control axon repulsion at the midline.

PubMed

Yang, Long; Bashaw, Greg J

2006-11-22

Son of sevenless (Sos) is a dual specificity guanine nucleotide exchange factor (GEF) that regulates both Ras and Rho family GTPases and thus is uniquely poised to integrate signals that affect both gene expression and cytoskeletal reorganization. Here, using genetics, biochemistry, and cell biology, we demonstrate that Sos is recruited to the plasma membrane, where it forms a ternary complex with the Roundabout receptor and the SH3-SH2 adaptor protein Dreadlocks (Dock) to regulate Rac-dependent cytoskeletal rearrangement in response to the Slit ligand. Intriguingly, the Ras and Rac-GEF activities of Sos can be uncoupled during Robo-mediated axon repulsion; Sos axon guidance function depends on its Rac-GEF activity, but not its Ras-GEF activity. These results provide in vivo evidence that the Ras and RhoGEF domains of Sos are separable signaling modules and support a model in which Robo recruits Sos to the membrane via Dock to activate Rac during midline repulsion.

The RhoA Activator GEF-H1/Lfc Is a Transforming Growth Factor-β Target Gene and Effector That Regulates α-Smooth Muscle Actin Expression and Cell Migration

PubMed Central

Tsapara, Anna; Luthert, Phillip; Greenwood, John; Hill, Caroline S.

2010-01-01

Maintenance of the epithelial phenotype is crucial for tissue homeostasis. In the retina, dedifferentiation and loss of integrity of the retinal pigment epithelium (RPE) leads to retinal dysfunction and fibrosis. Transforming growth factor (TGF)-β critically contributes to RPE dedifferentiation and induces various responses, including increased Rho signaling, up-regulation of α-smooth muscle actin (SMA), and cell migration and dedifferentiation. Cellular TGF-β responses are stimulated by different signal transduction pathways: some are Smad dependent and others Smad independent. Alterations in Rho signaling are crucial to both types of TGF-β signaling, but how TGF-β-stimulates Rho signaling is poorly understood. Here, we show that primary RPE cells up-regulated GEF-H1 in response to TGF-β. GEF-H1 was the only detectable Rho exchange factor increased by TGF-β1 in a genome-wide expression analysis. GEF-H1 induction was Smad4-dependant and led to Rho activation. GEF-H1 inhibition counteracted α-SMA up-regulation and cell migration. In patients with retinal detachments and fibrosis, migratory RPE cells exhibited increased GEF-H1 expression, indicating that induction occurs in diseased RPE in vivo. Our data indicate that GEF-H1 is a target and functional effector of TGF-β by orchestrating Rho signaling to regulate gene expression and cell migration, suggesting that it represents a new marker and possible therapeutic target for degenerative and fibrotic diseases. PMID:20089843

Why a Global International Waters Assessment (GIWA)?

PubMed

Hempel, Gotthilf; Daler, Dag

2004-02-01

Why GIWA? Six years ago several people had their doubts as to whether a Global International Waters Assessment would be worth the money and effort. Nowadays, it is no longer necessary to justify the creation of GIWA. On the contrary, we will show how important it was that the Global Environmental Facility (GEF) and UNEP, constituted GIWA. Countless water-related assessments focus on specific regions and/or specific issues. But GIWA is unique in its global and holistic policy-oriented approach applying a common methodology to address the major problems in all parts of the global hydrosphere. One major achievement of GIWA will be the GIWA publications which provide advice to GEF and other decision-making organizations. Further assets include the network of regional focal points and teams. GIWA encompasses marine, surface freshwater, and groundwater systems, following the flow of water from the sources in the mountains through the rivers and estuaries into the coastal waters and the shelf seas. GIWA studies the physical, chemical and biological properties of those waterbodies and living resources in relation to the human activities, combining ecological and socioeconomic considerations.

Robuste Verzweigungserkennung von Gefäßen in CTA-Datensätzen zur modellbasierten Extraktion der Centerline

NASA Astrophysics Data System (ADS)

Beck, Thomas; Fritz, Dominik; Biermann, Christina; Dillmann, Rüdiger

Bei der Befundung und Visualisierung von Blutgefäßen ist deren Centerline von zentraler Bedeutung. Die Unterscheidung zwischen unverzweigten Abschnitten des Gefäßes und Verzweigungsbereichen ermöglicht den Einsatz spezialisierter und sehr effizienter Algorithmen zur modellbasierten Extraktion der Centerline. In diesem Artikel wird ein robustes Verfahren zur Verzweigungserkennung vorgestellt. Das Verfahren beruht auf einem Front-Propagation-Ansatz mit dynamisch angepassten Schwellwerten und einer anschließenden Clusteranalyse. Die vorgestellte Methode zur Verzweigungserkennung wurde als Komponente einer Architektur zur Extraktion der Centerline auf handannotierten Datensätzen getestet. Erste Ergebnisse sind sehr vielversprechend und ermöglichen auch bei pathologischen Gefäßen eine robuste Detektion von Gefäßverzweigungen.

The C-terminus of the oncoprotein TGAT is necessary for plasma membrane association and efficient RhoA-mediated signaling.

PubMed

van Unen, J; Botman, D; Yin, T; Wu, Y I; Hink, M A; Gadella, T W J; Postma, M; Goedhart, J

2018-06-07

Rho guanine exchange factors (RhoGEFs) control cellular processes such as migration, adhesion and proliferation. Alternative splicing of the RhoGEF Trio produces TGAT. The RhoGEF TGAT is an oncoprotein with constitutive RhoGEF activity. We investigated whether the subcellular location of TGAT is critical for its RhoGEF activity. Since plasma membrane associated RhoGEFs are particularly effective at activating RhoA, plasma membrane localization of TGAT was examined. To this end, we developed a highly sensitive image analysis method to quantitatively measure plasma membrane association. The method requires a cytoplasmic marker and a plasma membrane marker, which are co-imaged with the tagged protein of interest. Linear unmixing is performed to determine the plasma membrane and cytoplasmic component in the fluorescence signal of protein of interest. The analysis revealed that wild-type TGAT is partially co-localized with the plasma membrane. Strikingly, cysteine TGAT-mutants lacking one or more putative palmitoylation sites in the C-tail, still showed membrane association. In contrast, a truncated variant, lacking the last 15 amino acids, TGAT Δ15 , lost membrane association. We show that membrane localization of TGAT was responsible for high RhoGEF activity by using a RhoA FRET-sensor and by determining F-actin levels. Mutants of TGAT that still maintained membrane association showed similar activity as wild-type TGAT. In contrast, the activity was abrogated for the cytoplasmic TGAT Δ15 variant. Synthetic recruitment of TGAT Δ15 to membranes confirmed that TGAT effectively activates RhoA at the plasma membrane. Together, these results show that membrane association of TGAT is critical for its activity.

AKAP13 Rho-GEF and PKD-Binding Domain Deficient Mice Develop Normally but Have an Abnormal Response to β-Adrenergic-Induced Cardiac Hypertrophy

PubMed Central

Spindler, Matthew J.; Burmeister, Brian T.; Huang, Yu; Hsiao, Edward C.; Salomonis, Nathan; Scott, Mark J.; Srivastava, Deepak; Carnegie, Graeme K.; Conklin, Bruce R.

2013-01-01

Background A-kinase anchoring proteins (AKAPs) are scaffolding molecules that coordinate and integrate G-protein signaling events to regulate development, physiology, and disease. One family member, AKAP13, encodes for multiple protein isoforms that contain binding sites for protein kinase A (PKA) and D (PKD) and an active Rho-guanine nucleotide exchange factor (Rho-GEF) domain. In mice, AKAP13 is required for development as null embryos die by embryonic day 10.5 with cardiovascular phenotypes. Additionally, the AKAP13 Rho-GEF and PKD-binding domains mediate cardiomyocyte hypertrophy in cell culture. However, the requirements for the Rho-GEF and PKD-binding domains during development and cardiac hypertrophy are unknown. Methodology/Principal Findings To determine if these AKAP13 protein domains are required for development, we used gene-trap events to create mutant mice that lacked the Rho-GEF and/or the protein kinase D-binding domains. Surprisingly, heterozygous matings produced mutant mice at Mendelian ratios that had normal viability and fertility. The adult mutant mice also had normal cardiac structure and electrocardiograms. To determine the role of these domains during β-adrenergic-induced cardiac hypertrophy, we stressed the mice with isoproterenol. We found that heart size was increased similarly in mice lacking the Rho-GEF and PKD-binding domains and wild-type controls. However, the mutant hearts had abnormal cardiac contractility as measured by fractional shortening and ejection fraction. Conclusions These results indicate that the Rho-GEF and PKD-binding domains of AKAP13 are not required for mouse development, normal cardiac architecture, or β-adrenergic-induced cardiac hypertrophic remodeling. However, these domains regulate aspects of β-adrenergic-induced cardiac hypertrophy. PMID:23658642

Characterization of the activation of small GTPases by their GEFs on membranes using artificial membrane tethering.

PubMed

Peurois, François; Veyron, Simon; Ferrandez, Yann; Ladid, Ilham; Benabdi, Sarah; Zeghouf, Mahel; Peyroche, Gérald; Cherfils, Jacqueline

2017-03-23

Active, GTP-bound small GTPases need to be attached to membranes by post-translational lipid modifications in order to process and propagate information in cells. However, generating and manipulating lipidated GTPases has remained difficult, which has limited our quantitative understanding of their activation by guanine nucleotide exchange factors (GEFs) and their termination by GTPase-activating proteins. Here, we replaced the lipid modification by a histidine tag in 11 full-length, human small GTPases belonging to the Arf, Rho and Rab families, which allowed to tether them to nickel-lipid-containing membranes and characterize the kinetics of their activation by GEFs. Remarkably, this strategy uncovered large effects of membranes on the efficiency and/or specificity in all systems studied. Notably, it recapitulated the release of autoinhibition of Arf1, Arf3, Arf4, Arf5 and Arf6 GTPases by membranes and revealed that all isoforms are efficiently activated by two GEFs with different regulatory regimes, ARNO and Brag2. It demonstrated that membranes stimulate the GEF activity of Trio toward RhoG by ∼30 fold and Rac1 by ∼10 fold, and uncovered a previously unknown broader specificity toward RhoA and Cdc42 that was undetectable in solution. Finally, it demonstrated that the exceptional affinity of the bacterial RabGEF DrrA for the phosphoinositide PI(4)P delimits the activation of Rab1 to the immediate vicinity of the membrane-bound GEF. Our study thus validates the histidine-tag strategy as a potent and simple means to mimic small GTPase lipidation, which opens a variety of applications to uncover regulations brought about by membranes. © 2017 The Author(s); published by Portland Press Limited on behalf of the Biochemical Society.

Trio’s Rho-specific GEF domain is the missing Gαq effector in C. elegans

PubMed Central

Williams, Stacey L.; Lutz, Susanne; Charlie, Nicole K.; Vettel, Christiane; Ailion, Michael; Coco, Cassandra; Tesmer, John J.G.; Jorgensen, Erik M.; Wieland, Thomas; Miller, Kenneth G.

2007-01-01

The Gαq pathway is essential for animal life and is a central pathway for driving locomotion, egg laying, and growth in Caenorhabditis elegans, where it exerts its effects through EGL-8 (phospholipase Cβ [PLCβ]) and at least one other effector. To find the missing effector, we performed forward genetic screens to suppress the slow growth and hyperactive behaviors of mutants with an overactive Gαq pathway. Four suppressor mutations disrupted the Rho-specific guanine-nucleotide exchange factor (GEF) domain of UNC-73 (Trio). The mutations produce defects in neuronal function, but not neuronal development, that cause sluggish locomotion similar to animals lacking EGL-8 (PLCβ). Strains containing null mutations in both EGL-8 (PLCβ) and UNC-73 (Trio RhoGEF) have strong synthetic phenotypes that phenocopy the arrested growth and near-complete paralysis of Gαq-null mutants. Using cell-based and biochemical assays, we show that activated C. elegans Gαq synergizes with Trio RhoGEF to activate RhoA. Activated Gαq and Trio RhoGEF appear to be part of a signaling complex, because they coimmunoprecipitate when expressed together in cells. Our results show that Trio’s Rho-specific GEF domain is a major Gαq effector that, together with PLCβ, mediates the Gαq signaling that drives the locomotion, egg laying, and growth of the animal. PMID:17942708

Diverse roles of guanine nucleotide exchange factors in regulating collective cell migration

PubMed Central

Tseng, Yun-Yu; Rabadán, M. Angeles; Krishna, Shefali; Hall, Alan

2017-01-01

Efficient collective migration depends on a balance between contractility and cytoskeletal rearrangements, adhesion, and mechanical cell–cell communication, all controlled by GTPases of the RHO family. By comprehensive screening of guanine nucleotide exchange factors (GEFs) in human bronchial epithelial cell monolayers, we identified GEFs that are required for collective migration at large, such as SOS1 and β-PIX, and RHOA GEFs that are implicated in intercellular communication. Down-regulation of the latter GEFs differentially enhanced front-to-back propagation of guidance cues through the monolayer and was mirrored by down-regulation of RHOA expression and myosin II activity. Phenotype-based clustering of knockdown behaviors identified RHOA-ARHGEF18 and ARHGEF3-ARHGEF28-ARHGEF11 clusters, indicating that the latter may signal through other RHO-family GTPases. Indeed, knockdown of RHOC produced an intermediate between the two phenotypes. We conclude that for effective collective migration, the RHOA-GEFs → RHOA/C → actomyosin pathways must be optimally tuned to compromise between generation of motility forces and restriction of intercellular communication. PMID:28512143

The General Environment Fit Scale: A Factor Analysis and Test of Convergent Construct Validity

PubMed Central

Beasley, Christopher; Jason, Leonard; Miller, Steven

2014-01-01

Person-environment fit (P-E fit) was initially espoused as an important construct in the field of community psychology; however, most of the theoretical and empirical development of the construct has been conducted by industrial/organizational (I/O) psychologists. In the current study, the GEFS—a P-E fit measure was developed from I/O and business management perspectives on fit—was administered to 246 attendees of an annual convention for residents and alumni of Oxford House (OH), a network of over 1400 mutual-help recovery homes. The authors conducted confirmatory factor and convergent construct validity analyses on the GEFS. The results suggested that the theoretical factor structure of the measure adequately fit the data, suggesting that the GEFS is a valid measure of P-E fit. OH resident fit with their recovery home was related to satisfaction, but not expected tenure. Exploratory analyses revealed that the sufficient supply of resident needs by the recovery home and similarity between residents and their housemates predicted satisfaction with the recovery home, but only similarity with housemates predicted how long residents intended to stay in the OHs. PMID:22071911

A KRAS GTPase K104Q Mutant Retains Downstream Signaling by Offsetting Defects in Regulation*

PubMed Central

Kistler, Samantha; George, Samuel D.; Kuhlmann, Nora; Garvey, Leslie; Huynh, Minh; Bagni, Rachel K.; Lammers, Michael; Der, Channing J.; Campbell, Sharon L.

2017-01-01

The KRAS GTPase plays a critical role in the control of cellular growth. The activity of KRAS is regulated by guanine nucleotide exchange factors (GEFs), GTPase-activating proteins (GAPs), and also post-translational modification. Lysine 104 in KRAS can be modified by ubiquitylation and acetylation, but the role of this residue in intrinsic KRAS function has not been well characterized. We find that lysine 104 is important for GEF recognition, because mutations at this position impaired GEF-mediated nucleotide exchange. Because the KRAS K104Q mutant has recently been employed as an acetylation mimetic, we conducted a series of studies to evaluate its in vitro and cell-based properties. Herein, we found that KRAS K104Q exhibited defects in both GEF-mediated exchange and GAP-mediated GTP hydrolysis, consistent with NMR-detected structural perturbations in localized regions of KRAS important for recognition of these regulatory proteins. Despite the partial defect in both GEF and GAP regulation, KRAS K104Q did not alter steady-state GTP-bound levels or the ability of the oncogenic KRAS G12V mutant to cause morphologic transformation of NIH 3T3 mouse fibroblasts and of WT KRAS to rescue the growth defect of mouse embryonic fibroblasts deficient in all Ras genes. We conclude that the KRAS K104Q mutant retains both WT and mutant KRAS function, probably due to offsetting defects in recognition of factors that up-regulate (GEF) and down-regulate (GAP) RAS activity. PMID:28154176

Novel Role for p21-activated Kinase 2 in Thrombin-induced Monocyte Migration*

PubMed Central

Gadepalli, Ravisekhar; Kotla, Sivareddy; Heckle, Mark R.; Verma, Shailendra K.; Singh, Nikhlesh K.; Rao, Gadiparthi N.

2013-01-01

To understand the role of thrombin in inflammation, we tested its effects on migration of THP-1 cells, a human monocytic cell line. Thrombin induced THP-1 cell migration in a dose-dependent manner. Thrombin induced tyrosine phosphorylation of Pyk2, Gab1, and p115 RhoGEF, leading to Rac1- and RhoA-dependent Pak2 activation. Downstream to Pyk2, Gab1 formed a complex with p115 RhoGEF involving their pleckstrin homology domains. Furthermore, inhibition or depletion of Pyk2, Gab1, p115 RhoGEF, Rac1, RhoA, or Pak2 levels substantially attenuated thrombin-induced THP-1 cell F-actin cytoskeletal remodeling and migration. Inhibition or depletion of PAR1 also blocked thrombin-induced activation of Pyk2, Gab1, p115 RhoGEF, Rac1, RhoA, and Pak2, resulting in diminished THP-1 cell F-actin cytoskeletal remodeling and migration. Similarly, depletion of Gα12 negated thrombin-induced Pyk2, Gab1, p115 RhoGEF, Rac1, RhoA, and Pak2 activation, leading to attenuation of THP-1 cell F-actin cytoskeletal remodeling and migration. These novel observations reveal that thrombin induces monocyte/macrophage migration via PAR1-Gα12-dependent Pyk2-mediated Gab1 and p115 RhoGEF interactions, leading to Rac1- and RhoA-targeted Pak2 activation. Thus, these findings provide mechanistic evidence for the role of thrombin and its receptor PAR1 in inflammation. PMID:24025335

Structure of Shigella IpgB2 in Complex with Human RhoA

PubMed Central

Klink, Björn U.; Barden, Stephan; Heidler, Thomas V.; Borchers, Christina; Ladwein, Markus; Stradal, Theresia E. B.; Rottner, Klemens; Heinz, Dirk W.

2010-01-01

A common theme in bacterial pathogenesis is the manipulation of eukaryotic cells by targeting the cytoskeleton. This is in most cases achieved either by modifying actin, or indirectly via activation of key regulators controlling actin dynamics such as Rho-GTPases. A novel group of bacterial virulence factors termed the WXXXE family has emerged as guanine nucleotide exchange factors (GEFs) for these GTPases. The precise mechanism of nucleotide exchange, however, has remained unclear. Here we report the structure of the WXXXE-protein IpgB2 from Shigella flexneri and its complex with human RhoA. We unambiguously identify IpgB2 as a bacterial RhoA-GEF and dissect the molecular mechanism of GDP release, an essential prerequisite for GTP binding. Our observations uncover that IpgB2 induces conformational changes on RhoA mimicking DbI- but not DOCK family GEFs. We also show that dissociation of the GDP·Mg2+ complex is preceded by the displacement of the metal ion to the α-phosphate of the nucleotide, diminishing its affinity to the GTPase. These data refine our understanding of the mode of action not only of WXXXE GEFs but also of mammalian GEFs of the DH/PH family. PMID:20363740

NCEP ENSEMBLE PRODUCTS

Science.gov Websites

://www.emc.ncep.noaa.gov/GEFS/ The links to the various Forecast Plots are listed under Experimental Data on the new GEFS page. This NCEP Ensemble Home Page is a collection of experimental analysis and forecast products

Vav1-mediated scaffolding interactions stabilize SLP-76 microclusters and contribute to antigen-dependent T cell responses.

PubMed

Sylvain, Nicholas R; Nguyen, Ken; Bunnell, Stephen C

2011-03-08

The guanine nucleotide exchange factor (GEF) Vav1 synergizes with the adaptor protein SLP-76 (Src homology 2 domain--containing leukocyte phosphoprotein of 76 kD) to support T cell development and activation. In response to ligation of the T cell receptor (TCR), SLP-76 is assembled into microclusters that provide an essential platform for the signaling events that drive T cell activation. We found that Vav1 selectively entered SLP-76 microclusters, rather than TCR microclusters, influencing their stability and function. The carboxyl terminus of Vav1, which consists of Src homology domains, was both necessary and sufficient for the entry of Vav1 into SLP-76 microclusters; however, this fragment of Vav1 was insufficient to stabilize the microclusters, and it potently suppressed T cell activation. This indicated that the amino terminus of Vav1, which has the GEF domain, also contributed to the integrity of SLP-76 microclusters and thereby to T cell activation. These microcluster-stabilizing functions were independent of the GEF activity in the amino terminus of Vav1 and were unaffected if the GEF function of Vav1 was either inactivated or constitutively activated by mutation. In contrast, Vav1 deletion mutants lacking either the calponin homology domain or the catalytic core of the GEF exhibited mild scaffolding defects, but they differentially affected TCR-dependent calcium ion (Ca²+) responses. We conclude that multiple GEF-independent scaffolding functions distributed throughout the amino terminus of Vav1 contribute to the activation of T cells by acting synergistically to increase the stability and function of SLP-76 microclusters.

Agonist-induced Ca2+ Sensitization in Smooth Muscle

PubMed Central

Artamonov, Mykhaylo V.; Momotani, Ko; Stevenson, Andra; Trentham, David R.; Derewenda, Urszula; Derewenda, Zygmunt S.; Read, Paul W.; Gutkind, J. Silvio; Somlyo, Avril V.

2013-01-01

Many agonists, acting through G-protein-coupled receptors and Gα subunits of the heterotrimeric G-proteins, induce contraction of smooth muscle through an increase of [Ca2+]i as well as activation of the RhoA/RhoA-activated kinase pathway that amplifies the contractile force, a phenomenon known as Ca2+ sensitization. Gα12/13 subunits are known to activate the regulator of G-protein signaling-like family of guanine nucleotide exchange factors (RhoGEFs), which includes PDZ-RhoGEF (PRG) and leukemia-associated RhoGEF (LARG). However, their contributions to Ca2+-sensitized force are not well understood. Using permeabilized blood vessels from PRG(−/−) mice and a new method to silence LARG in organ-cultured blood vessels, we show that both RhoGEFs are activated by the physiologically and pathophysiologically important thromboxane A2 and endothelin-1 receptors. The co-activation is the result of direct and independent activation of both RhoGEFs as well as their co-recruitment due to heterodimerization. The isolated recombinant C-terminal domain of PRG, which is responsible for heterodimerization with LARG, strongly inhibited Ca2+-sensitized force. We used photolysis of caged phenylephrine, caged guanosine 5′-O-(thiotriphosphate) (GTPγS) in solution, and caged GTPγS or caged GTP loaded on the RhoA·RhoGDI complex to show that the recruitment and activation of RhoGEFs is the cause of a significant time lag between the initial Ca2+ transient and phasic force components and the onset of Ca2+-sensitized force. PMID:24106280

RHGF-2 Is an Essential Rho-1 Specific RhoGEF that binds to the Multi-PDZ Domain Scaffold Protein MPZ-1 in Caenorhabditis elegans

PubMed Central

Lin, Li; Tran, Thuy; Hu, Shuang; Cramer, Todd; Komuniecki, Richard; Steven, Robert M.

2012-01-01

RhoGEF proteins activate the Rho family of small GTPases and thus play a key role in regulating fundamental cellular processes such as cell morphology and polarity, cell cycle progression and gene transcription. We identified a Caenorhabditis elegans RhoGEF protein, RHGF-2, as a binding partner of the C. elegans multi-PDZ domain scaffold protein MPZ-1 (MUPP1 in mammals). RHGF-2 exhibits significant identity to the mammalian RhoGEFs PLEKHG5/Tech/Syx and contains a class I C-terminal PDZ binding motif (SDV) that interacts most strongly to MPZ-1 PDZ domain eight. RHGF-2 RhoGEF activity is specific to the C. elegans RhoA homolog RHO-1 as determined by direct binding, GDP/GTP exchange and serum response element-driven reporter activity. rhgf-2 is an essential gene since rhgf-2 deletion mutants do not elongate during embryogenesis and hatch as short immobile animals that arrest development. Interestingly, the expression of a functional rhgf-2::gfp transgene appears to be exclusively neuronal and rhgf-2 overexpression results in loopy movement with exaggerated body bends. Transient expression of RHGF-2 in N1E-115 neuroblastoma cells prevents neurite outgrowth similar to constitutive RhoA activation in these cells. Together, these observations indicate neuronally expressed RHGF-2 is an essential RHO-1 specific RhoGEF that binds most strongly to MPZ-1 PDZ domain eight and is required for wild-type C. elegans morphology and growth. PMID:22363657

A KRAS GTPase K104Q Mutant Retains Downstream Signaling by Offsetting Defects in Regulation.

PubMed

Yin, Guowei; Kistler, Samantha; George, Samuel D; Kuhlmann, Nora; Garvey, Leslie; Huynh, Minh; Bagni, Rachel K; Lammers, Michael; Der, Channing J; Campbell, Sharon L

2017-03-17

The KRAS GTPase plays a critical role in the control of cellular growth. The activity of KRAS is regulated by guanine nucleotide exchange factors (GEFs), GTPase-activating proteins (GAPs), and also post-translational modification. Lysine 104 in KRAS can be modified by ubiquitylation and acetylation, but the role of this residue in intrinsic KRAS function has not been well characterized. We find that lysine 104 is important for GEF recognition, because mutations at this position impaired GEF-mediated nucleotide exchange. Because the KRAS K104Q mutant has recently been employed as an acetylation mimetic, we conducted a series of studies to evaluate its in vitro and cell-based properties. Herein, we found that KRAS K104Q exhibited defects in both GEF-mediated exchange and GAP-mediated GTP hydrolysis, consistent with NMR-detected structural perturbations in localized regions of KRAS important for recognition of these regulatory proteins. Despite the partial defect in both GEF and GAP regulation, KRAS K104Q did not alter steady-state GTP-bound levels or the ability of the oncogenic KRAS G12V mutant to cause morphologic transformation of NIH 3T3 mouse fibroblasts and of WT KRAS to rescue the growth defect of mouse embryonic fibroblasts deficient in all Ras genes. We conclude that the KRAS K104Q mutant retains both WT and mutant KRAS function, probably due to offsetting defects in recognition of factors that up-regulate (GEF) and down-regulate (GAP) RAS activity. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Quantitative Analysis of Guanine Nucleotide Exchange Factors (GEFs) as Enzymes

PubMed Central

Randazzo, Paul A; Jian, Xiaoying; Chen, Pei-Wen; Zhai, Peng; Soubias, Olivier; Northup, John K

2014-01-01

The proteins that possess guanine nucleotide exchange factor (GEF) activity, which include about ~800 G protein coupled receptors (GPCRs),1 15 Arf GEFs,2 81 Rho GEFs,3 8 Ras GEFs,4 and others for other families of GTPases,5 catalyze the exchange of GTP for GDP on all regulatory guanine nucleotide binding proteins. Despite their importance as catalysts, relatively few exchange factors (we are aware of only eight for ras superfamily members) have been rigorously characterized kinetically.5–13 In some cases, kinetic analysis has been simplistic leading to erroneous conclusions about mechanism (as discussed in a recent review14). In this paper, we compare two approaches for determining the kinetic properties of exchange factors: (i) examining individual equilibria, and; (ii) analyzing the exchange factors as enzymes. Each approach, when thoughtfully used,14,15 provides important mechanistic information about the exchange factors. The analysis as enzymes is described in further detail. With the focus on the production of the biologically relevant guanine nucleotide binding protein complexed with GTP (G•GTP), we believe it is conceptually simpler to connect the kinetic properties to cellular effects. Further, the experiments are often more tractable than those used to analyze the equilibrium system and, therefore, more widely accessible to scientists interested in the function of exchange factors. PMID:25332840

Structural Basis of Membrane Targeting by the Dock180 Family of Rho Family Guanine Exchange Factors (Rho-GEFs)*

PubMed Central

Premkumar, Lakshmanane; Bobkov, Andrey A.; Patel, Manishha; Jaroszewski, Lukasz; Bankston, Laurie A.; Stec, Boguslaw; Vuori, Kristiina; Côté, Jean-Francois; Liddington, Robert C.

2010-01-01

The Dock180 family of atypical Rho family guanine nucleotide exchange factors (Rho-GEFs) regulate a variety of processes involving cellular or subcellular polarization, including cell migration and phagocytosis. Each contains a Dock homology region-1 (DHR-1) domain that is required to localize its GEF activity to a specific membrane compartment where levels of phosphatidylinositol (3,4,5)-trisphosphate (PtdIns(3,4,5)P3) are up-regulated by the local activity of PtdIns 3-kinase. Here we define the structural and energetic bases of phosphoinositide specificity by the DHR-1 domain of Dock1 (a GEF for Rac1), and show that DHR-1 utilizes a C2 domain scaffold and surface loops to create a basic pocket on its upper surface for recognition of the PtdIns(3,4,5)P3 head group. The pocket has many of the characteristics of those observed in pleckstrin homology domains. We show that point mutations in the pocket that abolish phospholipid binding in vitro ablate the ability of Dock1 to induce cell polarization, and propose a model that brings together recent mechanistic and structural studies to rationalize the central role of DHR-1 in dynamic membrane targeting of the Rho-GEF activity of Dock180. PMID:20167601

The Transboundary Waters Assessment Programme (TWAP) River Basin Component Methods and Results

NASA Astrophysics Data System (ADS)

de Sherbinin, A. M.; Glennie, P.

2014-12-01

The Transboundary Waters Assessment Programme (TWAP) was initiated by the Global Environment Facility (GEF) to create the first baseline assessment of all of the planet's transboundary water resources. The TWAP River Basin component consists of a baseline comparative assessment of 270 transboundary river basins, including all but the smallest basins, to enable the identification of priority issues and hotspots at risk from a variety of stressors. The assessment is indicator based and it is intended to provide a relative analysis of basins based on risks to societies and ecosystems. Models and observational data have been used to create 14 indicators covering environmental, human and agricultural water stress; nutrient and wastewater pollution; extinction risk; governance and institutions; economic dependence on water resources; societal wellbeing at sub-basin scales; and societal risks from climate extremes. The methodology is not limited to transboundary basins, but can be applied to all river basins. This presentation will provide a summary of the methods and results of the TWAP River Basin component. It will also briefly discuss preliminary results of the TWAP lakes and aquifer components.

Binational collaboration to study Gulf of Mexico's harmful algae

NASA Astrophysics Data System (ADS)

Soto, Inia; Hu, Chuanmin; Steidinger, Karen; Muller-Karger, Frank; Cannizzaro, Jennifer; Wolny, Jennifer; Cerdeira-Estrada, Sergio; Santamaria-del-Angel, Eduardo; Tafoya-del-Angel, Fausto; Alvarez-Torres, Porfirio; Herrera Silveira, Jorge; Allen, Jeanne

2012-01-01

Blooms of the toxic marine dinoflagellate Karenia brevis cause massive fish kills and other public health and economic problems in coastal waters throughout the Gulf of Mexico [Steidinger, 2009]. These harmful algal blooms (HABs) are a gulf-wide problem that require a synoptic observing system for better serving decision-making needs. The major nutrient sources that initiate and maintain these HABs and the possible connectivity of blooms in different locations are important questions being addressed through new collaborations between Mexican and U.S. researchers and government institutions. These efforts were originally organized under the U.S./Mexico binational partnership for the HABs Observing System (HABSOS), led by the U.S. Environmental Protection Agency's Gulf of Mexico Program (EPAGMP) and several agencies in Veracruz, Mexico, since 2006. In 2010 these efforts were expanded to include other Mexican states and institutions with the integrated assessment and management of the Gulf of Mexico Large Marine Ecosystem (GoMLME) program sponsored by the Global Environment Facility (GEF), the United Nations Industrial Development Organization (UNIDO), the Secretaría de Medio Ambiente y Recursos Naturales (SEMARNAT), and the National Oceanic and Atmospheric Administration (NOAA).

Choosing to Win: How Sof Can Better Select Partners for Capacity Building

DTIC Science & Technology

2014-06-01

countries will yield the best results when SOF are employed to build capacity. This thesis uses two RAND reports—What Works Best When Building...environment, planners will be forced to make difficult decisions about which countries will yield the best results when SOF are employed to build...deliberate planning, global posture, global force management, and nuclear weapons planning.20 The GEF also directs the combatant commanders to

The Large Marine Ecosystem Approach for 21st Century Ocean Health and International Sustainable Development

NASA Astrophysics Data System (ADS)

Honey, K. T.

2014-12-01

The global coastal ocean and watersheds are divided into 66 Large Marine Ecosystems (LMEs), which encompass regions from river basins, estuaries, and coasts to the seaward boundaries of continental shelves and margins of major currents. Approximately 80% of global fisheries catch comes from LME waters. Ecosystem goods and services from LMEs contribute an estimated US 18-25 trillion dollars annually to the global economy in market and non-market value. The critical importance of these large-scale systems, however, is threatened by human populations and pressures, including climate change. Fortunately, there is pragmatic reason for optimism. Interdisciplinary frameworks exist, such as the Large Marine Ecosystem (LME) approach for adaptive management that can integrate both nature-centric and human-centric views into ecosystem monitoring, assessment, and adaptive management practices for long-term sustainability. Originally proposed almost 30 years ago, the LME approach rests on five modules are: (i) productivity, (ii) fish and fisheries, (iii) pollution and ecosystem health, (iv) socioeconomics, and (v) governance for iterative adaptive management at a large, international scale of 200,000 km2 or greater. The Global Environment Facility (GEF), World Bank, and United Nations agencies recognize and support the LME approach—as evidenced by over 3.15 billion in financial assistance to date for LME projects. This year of 2014 is an exciting milestone in LME history, after 20 years of the United Nations and GEF organizations adopting LMEs as a unit for ecosystem-based approaches to management. The LME approach, however, is not perfect. Nor is it immutable. Similar to the adaptive management framework it propones, the LME approach itself must adapt to new and emerging 21st Century technologies, science, and realities. The LME approach must further consider socioeconomics and governance. Within the socioeconomics module alone, several trillion-dollar opportunities exist for interdisciplinary integration with best practices in: (i) water-energy nexus infrastructure; (ii) responsible tourism; and (iii) open data innovations.

The G Protein α Chaperone Ric-8 as a Potential Therapeutic Target

PubMed Central

Papasergi, Makaía M.; Patel, Bharti R.

2015-01-01

Resistance to inhibitors of cholinesterase (Ric-8)A and Ric-8B are essential genes that encode positive regulators of heterotrimeric G protein α subunits. Controversy persists surrounding the precise way(s) that Ric-8 proteins affect G protein biology and signaling. Ric-8 proteins chaperone nucleotide-free Gα-subunit states during biosynthetic protein folding prior to G protein heterotrimer assembly. In organisms spanning the evolutionary window of Ric-8 expression, experimental perturbation of Ric-8 genes results in reduced functional abundances of G proteins because G protein α subunits are misfolded and degraded rapidly. Ric-8 proteins also act as Gα-subunit guanine nucleotide exchange factors (GEFs) in vitro. However, Ric-8 GEF activity could strictly be an in vitro phenomenon stemming from the ability of Ric-8 to induce partial Gα unfolding, thereby enhancing GDP release. Ric-8 GEF activity clearly differs from the GEF activity of G protein–coupled receptors (GPCRs). G protein βγ is inhibitory to Ric-8 action but obligate for receptors. It remains an open question whether Ric-8 has dual functions in cells and regulates G proteins as both a molecular chaperone and GEF. Clearly, Ric-8 has a profound influence on heterotrimeric G protein function. For this reason, we propose that Ric-8 proteins are as yet untested therapeutic targets in which pharmacological inhibition of the Ric-8/Gα protein–protein interface could serve to attenuate the effects of disease-causing G proteins (constitutively active mutants) and/or GPCR signaling. This minireview will chronicle the understanding of Ric-8 function, provide a comparative discussion of the Ric-8 molecular chaperoning and GEF activities, and support the case for why Ric-8 proteins should be considered potential targets for development of new therapies. PMID:25319541

Assessing public health policy approaches to level-up the gradient in health inequalities: the Gradient Evaluation Framework.

PubMed

Davies, J K; Sherriff, N S

2014-03-01

This paper seeks to introduce and analyse the development of the Gradient Evaluation Framework (GEF) to facilitate evaluation of policy actions for their current or future use in terms of their 'gradient friendliness'. In particular, this means their potential to level-up the gradient in health inequalities by addressing the social determinants of health and thereby reducing decision-makers' chances of error when developing such policy actions. A qualitative developmental study to produce a policy-based evaluation framework. The scientific basis of GEF was developed using a comprehensive consensus-building process. This process followed an initial narrative review, based on realist review principles, which highlighted the need for production of a dedicated evaluation framework. The consensus-building process included expert workshops, a pretesting phase, and external peer review, together with support from the Gradient project Scientific Advisory Group and all Gradient project partners, including its Project Steering Committee. GEF is presented as a flexible policy tool resulting from a consensus-building process involving experts from 13 European countries. The theoretical foundations which underpin GEF are discussed, together with a range of practical challenges. The importance of systematic evaluation at each stage of the policy development and implementation cycle is highlighted, as well as the socio-political context in which policy actions are located. GEF offers potentially a major contribution to the public health field in the form of a practical, policy-relevant and common frame of reference for the evaluation of public health interventions that aim to level-up the social gradient in health inequalities. Further research, including the need for practical field testing of GEF and the exploration of alternative presentational formats, is recommended. Copyright © 2013 The Royal Society for Public Health. Published by Elsevier Ltd. All rights reserved.

National Centers for Environmental Prediction

Science.gov Websites

/ VISION | About EMC EMC > GEFS > OPERATIONAL PRODUCTS Home Operational Products Experimental Data Global Ensemble products posted by the GEFS model team are experimental analysis and forecast products , and you will find them listed under the "Experimental Data" section of our web site. The

A non-canonical role for Rgnef in promoting integrin-stimulated focal adhesion kinase activation

PubMed Central

Miller, Nichol L. G.; Lawson, Christine; Kleinschmidt, Elizabeth G.; Tancioni, Isabelle; Uryu, Sean; Schlaepfer, David D.

2013-01-01

Summary Rgnef (also known as p190RhoGEF or ARHGEF28) is a Rho guanine-nucleotide-exchange factor (GEF) that binds focal adhesion kinase (FAK). FAK is recruited to adhesions and activated by integrin receptors binding to matrix proteins, such as fibronectin (FN). Canonical models place Rgnef downstream of integrin–FAK signaling in regulating Rho GTPase activity and cell movement. Herein, we establish a new, upstream role for Rgnef in enhancing FAK localization to early peripheral adhesions and promoting FAK activation upon FN binding. Rgnef-null mouse embryo fibroblasts (MEFs) exhibit defects in adhesion formation, levels of FAK phosphotyrosine (pY)-397 and FAK localization to peripheral adhesions upon re-plating on FN. Rgnef re-expression rescues these defects, but requires Rgnef–FAK binding. A mutation in the Rgnef pleckstrin homology (PH) domain inhibits adhesion formation, FAK localization, and FAK-Y397 and paxillin-Y118 phosphorylation without disrupting the Rgnef–FAK interaction. A GEF-inactive Rgnef mutant rescues FAK-Y397 phosphorylation and early adhesion localization, but not paxillin-Y118 phosphorylation. This suggests that, downstream of FN binding, paxillin-pY118 requires Rgnef GEF activity through a mechanism distinct from adhesion formation and FAK activation. These results support a scaffolding role for Rgnef in FAK localization and activation at early adhesions in a PH-domain-dependent but GEF-activity-independent manner. PMID:24006257

A Steric-inhibition model for regulation of nucleotide exchange via the Dock180 family of GEFs.

PubMed

Lu, Mingjian; Kinchen, Jason M; Rossman, Kent L; Grimsley, Cynthia; Hall, Matthew; Sondek, John; Hengartner, Michael O; Yajnik, Vijay; Ravichandran, Kodi S

2005-02-22

CDM (CED-5, Dock180, Myoblast city) family members have been recently identified as novel, evolutionarily conserved guanine nucleotide exchange factors (GEFs) for Rho-family GTPases . They regulate multiple processes, including embryonic development, cell migration, apoptotic-cell engulfment, tumor invasion, and HIV-1 infection, in diverse model systems . However, the mechanism(s) of regulation of CDM proteins has not been well understood. Here, our studies on the prototype member Dock180 reveal a steric-inhibition model for regulating the Dock180 family of GEFs. At basal state, the N-terminal SH3 domain of Dock180 binds to the distant catalytic Docker domain and negatively regulates the function of Dock180. Further studies revealed that the SH3:Docker interaction sterically blocks Rac access to the Docker domain. Interestingly, ELMO binding to the SH3 domain of Dock180 disrupted the SH3:Docker interaction, facilitated Rac access to the Docker domain, and contributed to the GEF activity of the Dock180/ELMO complex. Additional genetic rescue studies in C. elegans suggested that the regulation of the Docker-domain-mediated GEF activity by the SH3 domain and its adjoining region is evolutionarily conserved. This steric-inhibition model may be a general mechanism for regulating multiple SH3-domain-containing Dock180 family members and may have implications for a variety of biological processes.

Deconstructing Ras Signaling in the Thymus

PubMed Central

Kortum, Robert L.; Sommers, Connie L.; Pinski, John M.; Alexander, Clayton P.; Merrill, Robert K.; Li, Wenmei; Love, Paul E.

2012-01-01

Thymocytes must transit at least two distinct developmental checkpoints, governed by signals that emanate from either the pre-T cell receptor (pre-TCR) or the TCR to the small G protein Ras before emerging as functional T lymphocytes. Recent studies have shown a role for the Ras guanine exchange factor (RasGEF) Sos1 at the pre-TCR checkpoint. At the second checkpoint, the quality of signaling through the TCR is interrogated to ensure the production of an appropriate T cell repertoire. Although RasGRP1 is the only confirmed RasGEF required at the TCR checkpoint, current models suggest that the intensity and character of Ras activation, facilitated by both Sos and RasGRP1, will govern the boundary between survival (positive selection) and death (negative selection) at this stage. Using mouse models, we have assessed the independent and combined roles for the RasGEFs Sos1, Sos2, and RasGRP1 during thymocyte development. Although Sos1 was the dominant RasGEF at the pre-TCR checkpoint, combined Sos1/RasGRP1 deletion was required to effectively block development at this stage. Conversely, while RasGRP1 deletion efficiently blocked positive selection, combined RasGRP1/Sos1 deletion was required to block negative selection. This functional redundancy in RasGEFs during negative selection may act as a failsafe mechanism ensuring appropriate central tolerance. PMID:22586275

The integrity of the plant Golgi apparatus depends on cell growth-controlled activity of GNL1.

PubMed

Du, Wenyan; Tamura, Kentaro; Stefano, Giovanni; Brandizzi, Federica

2013-05-01

Membrane traffic and organelle integrity in the plant secretory pathway depend on ARF-GTPases, which are activated by guanine-nucleotide exchange factors (ARF-GEFs). While maintenance of conserved roles, evolution of unique functions as well as tissue-specific roles have been shown for a handful of plant ARF-GEFs, a fundamental yet unanswered question concerns the extent to which their function overlaps during cell growth. To address this, we have characterized pao, a novel allele of GNOM-like 1 (GNL1), a brefeldin A (BFA)-insensitive ARF-GEF, isolated through a confocal microscopy-based forward genetics screen of the Golgi in Arabidopsis thaliana. Specifically, we have analyzed the dependence of the integrity of trafficking routes and secretory organelles on GNL1 availability during expansion stages of cotyledon epidermal cells, an exquisite model system for vegetative cell growth analyses in intact tissues. We show that Golgi traffic is influenced largely by GNL1 availability at early stages of cotyledon cell expansion but by BFA-sensitive GEFs when cell growth terminates. These data reveal an unanticipated level of complexity in the biology of GNL1 by showing that its cellular roles are correlated with cell growth. These results also indicate that the cell growth stage is an important element weighting into functional analyses of the cellular roles of ARF-GEFs.

Metabolomics reveals the formation of aldehydes and iminium in gefitinib metabolism

USDA-ARS?s Scientific Manuscript database

Gefitinib (GEF), an inhibitor of epidermal growth factor receptor (EGFR) tyrosine kinase, is widely used for the treatment of cancers, particularly non-small cell lung cancer. However, its clinical use is limited by multiple adverse effects associated with GEF, such as liver and lung injuries, sever...

Evaluation of ensemble precipitation forecasts generated through post-processing in a Canadian catchment

NASA Astrophysics Data System (ADS)

Jha, Sanjeev K.; Shrestha, Durga L.; Stadnyk, Tricia A.; Coulibaly, Paulin

2018-03-01

Flooding in Canada is often caused by heavy rainfall during the snowmelt period. Hydrologic forecast centers rely on precipitation forecasts obtained from numerical weather prediction (NWP) models to enforce hydrological models for streamflow forecasting. The uncertainties in raw quantitative precipitation forecasts (QPFs) are enhanced by physiography and orography effects over a diverse landscape, particularly in the western catchments of Canada. A Bayesian post-processing approach called rainfall post-processing (RPP), developed in Australia (Robertson et al., 2013; Shrestha et al., 2015), has been applied to assess its forecast performance in a Canadian catchment. Raw QPFs obtained from two sources, Global Ensemble Forecasting System (GEFS) Reforecast 2 project, from the National Centers for Environmental Prediction, and Global Deterministic Forecast System (GDPS), from Environment and Climate Change Canada, are used in this study. The study period from January 2013 to December 2015 covered a major flood event in Calgary, Alberta, Canada. Post-processed results show that the RPP is able to remove the bias and reduce the errors of both GEFS and GDPS forecasts. Ensembles generated from the RPP reliably quantify the forecast uncertainty.

Convex foundations for generalized MaxEnt models

NASA Astrophysics Data System (ADS)

Frongillo, Rafael; Reid, Mark D.

2014-12-01

We present an approach to maximum entropy models that highlights the convex geometry and duality of generalized exponential families (GEFs) and their connection to Bregman divergences. Using our framework, we are able to resolve a puzzling aspect of the bijection of Banerjee and coauthors between classical exponential families and what they call regular Bregman divergences. Their regularity condition rules out all but Bregman divergences generated from log-convex generators. We recover their bijection and show that a much broader class of divergences correspond to GEFs via two key observations: 1) Like classical exponential families, GEFs have a "cumulant" C whose subdifferential contains the mean: Eo˜pθ[φ(o)]∈∂C(θ) ; 2) Generalized relative entropy is a C-Bregman divergence between parameters: DF(pθ,pθ')= D C(θ,θ') , where DF becomes the KL divergence for F = -H. We also show that every incomplete market with cost function C can be expressed as a complete market, where the prices are constrained to be a GEF with cumulant C. This provides an entirely new interpretation of prediction markets, relating their design back to the principle of maximum entropy.

Dynamic Control of Excitatory Synapse Development by a Rac1 GEF/GAP Regulatory Complex

PubMed Central

Um, Kyongmi; Niu, Sanyong; Duman, Joseph G.; Cheng, Jinxuan; Tu, Yen-Kuei; Schwechter, Brandon; Liu, Feng; Hiles, Laura; Narayanan, Anjana; Ash, Ryan T.; Mulherkar, Shalaka; Alpadi, Kannan; Smirnakis, Stelios M.; Tolias, Kimberley F.

2014-01-01

SUMMARY The small GTPase Rac1 orchestrates actin-dependent remodeling essential for numerous cellular processes including synapse development. While precise spatiotemporal regulation of Rac1 is necessary for its function, little is known about the mechanisms that enable Rac1 activators (GEFs) and inhibitors (GAPs) to act in concert to regulate Rac1 signaling. Here we identify a regulatory complex composed of a Rac-GEF (Tiam1) and a Rac-GAP (Bcr) that cooperate to control excitatory synapse development. Disruption of Bcr function within this complex increases Rac1 activity and dendritic spine remodeling, resulting in excessive synaptic growth that is rescued by Tiam1 inhibition. Notably, EphB receptors utilize the Tiam1-Bcr complex to control synaptogenesis. Following EphB activation, Tiam1 induces Rac1-dependent spine formation, whereas Bcr prevents Rac1-mediated receptor internalization, promoting spine growth over retraction. The finding that a Rac-specific GEF/GAP complex is required to maintain optimal levels of Rac1 signaling provides an important insight into the regulation of small GTPases. PMID:24960694

Kin5 Knockdown in Tetrahymena thermophila Using RNAi Blocks Cargo Transport of Gef1

PubMed Central

Awan, Aashir; Bell, Aaron J.; Satir, Peter

2009-01-01

A critical process that builds and maintains the eukaryotic cilium is intraflagellar transport (IFT). This process utilizes members of the kinesin-2 superfamily to transport cargo into the cilium (anterograde transport) and a dynein motor for the retrograde traffic. Using a novel RNAi knockdown method, we have analyzed the function of the homodimeric IFT kinesin-2, Kin5, in Tetrahymena ciliary transport. In RNAi transformants, Kin5 was severely downregulated and disappeared from the cilia, but cilia did not resorb, although tip structure was affected. After deciliation of the knockdown cell, cilia regrew and cells swam, which suggested that Kin5 is not responsible for the trafficking of axonemal precursors to build the cilium, but could be transporting molecules that act in ciliary signal transduction, such as guanine nucleotide exchange proteins (GEFs). Gef1 is a Tetrahymena ciliary protein, and current coimmunoprecipitation and immunofluorescence studies showed that it is absent in regrowing cilia of the knockdown cells lacking ciliary Kin5. We suggest that one important cargo of Kin5 is Gef1 and knockdown of Kin5 results in cell lethality. PMID:19290045

Neurodevelopmental disease-associated de novo mutations and rare sequence variants affect TRIO GDP/GTP exchange factor activity.

PubMed

Katrancha, Sara M; Wu, Yi; Zhu, Minsheng; Eipper, Betty A; Koleske, Anthony J; Mains, Richard E

2017-12-01

Bipolar disorder, schizophrenia, autism and intellectual disability are complex neurodevelopmental disorders, debilitating millions of people. Therapeutic progress is limited by poor understanding of underlying molecular pathways. Using a targeted search, we identified an enrichment of de novo mutations in the gene encoding the 330-kDa triple functional domain (TRIO) protein associated with neurodevelopmental disorders. By generating multiple TRIO antibodies, we show that the smaller TRIO9 isoform is the major brain protein product, and its levels decrease after birth. TRIO9 contains two guanine nucleotide exchange factor (GEF) domains with distinct specificities: GEF1 activates both Rac1 and RhoG; GEF2 activates RhoA. To understand the impact of disease-associated de novo mutations and other rare sequence variants on TRIO function, we utilized two FRET-based biosensors: a Rac1 biosensor to study mutations in TRIO (T)GEF1, and a RhoA biosensor to study mutations in TGEF2. We discovered that one autism-associated de novo mutation in TGEF1 (K1431M), at the TGEF1/Rac1 interface, markedly decreased its overall activity toward Rac1. A schizophrenia-associated rare sequence variant in TGEF1 (F1538Intron) was substantially less active, normalized to protein level and expressed poorly. Overall, mutations in TGEF1 decreased GEF1 activity toward Rac1. One bipolar disorder-associated rare variant (M2145T) in TGEF2 impaired inhibition by the TGEF2 pleckstrin-homology domain, resulting in dramatically increased TGEF2 activity. Overall, genetic damage to both TGEF domains altered TRIO catalytic activity, decreasing TGEF1 activity and increasing TGEF2 activity. Importantly, both GEF changes are expected to decrease neurite outgrowth, perhaps consistent with their association with neurodevelopmental disorders. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Neuropeptide-stimulated cell migration in prostate cancer cells is mediated by RhoA kinase signaling and inhibited by neutral endopeptidase.

PubMed

Zheng, R; Iwase, A; Shen, R; Goodman, O B; Sugimoto, N; Takuwa, Y; Lerner, D J; Nanus, D M

2006-09-28

The neuropeptides bombesin and endothelin-1 stimulate prostate cancer (PC) cell migration and invasion (J Clin Invest, 2000; 106: 1399-1407). The intracellular signaling pathways that direct this cell movement are not well delineated. The monomeric GTPase RhoA is required for migration in several cell types including neutrophils, monocytes and fibroblasts. We demonstrate that bombesin-stimulated PC cell migration occurs via the heterotrimeric G-protein-coupled receptors (G-protein) G alpha 13 subunit leading to activation of RhoA, and Rho-associated coiled-coil forming protein kinase (ROCK). Using siRNA to suppress expression of the three known G-protein alpha-subunit-associated RhoA guanine nucleotide exchange factors (GEFs), we also show that two of these RhoA GEFs, PDZ-RhoGEF and leukemia-associated RhoGEF (LARG), link bombesin receptors to RhoA in a non-redundant manner in PC cells. We next show that focal adhesion kinase, which activates PDZ-RhoGEF and LARG, is required for bombesin-stimulated RhoA activation. Neutral endopeptidase (NEP) is expressed on normal prostate epithelium whereas loss of NEP expression contributes to PC progression. We also demonstrate that NEP inhibits neuropeptide activation of RhoA. Together, these results establish a contiguous signaling pathway from the bombesin receptor to ROCK in PC cells, and they implicate NEP as a major regulator of neuropeptide-stimulated RhoA in these cells. This work also identifies members of this signaling pathway as potential targets for rational pharmacologic manipulation of neuropeptide-stimulated migration of PC cells.

Factors Influencing Farmers' Willingness to Participate in the Conversion of Cultivated Land to Wetland Program in Sanjiang National Nature Reserve, China

NASA Astrophysics Data System (ADS)

Zhang, Chunli; Robinson, Daniel; Wang, Jing; Liu, Jibin; Liu, Xiaohui; Tong, Lianjun

2011-01-01

Sanjiang National Nature Reserve (NNR) is a state-owned natural wetland in China that has suffered severe degradation due to cultivation and wetland reclamation by farmers. As a consequence, the conversion of cultivated land to wetlands (CCW) was proposed by the government of Heilongjiang province and the United Nations Development Programme/Global Environment Facility (UNDP/GEF) project team in 2007. We suggest that voluntary participation in the CCW could be an important tool for accomplishing the integrated objectives of wetland conservation and local development. The purpose of this study was to examine the main factors that influence farmers' willingness to participate in the CCW through a field investigation and a questionnaire. Based on the data from our questionnaire, which provided an effective sample of 310 households in 11 villages, the influencing factors of farmers' willingness to participate were analyzed through binary logistic regression analyses. It was concluded that age, education, the amount of cultivated land, geographical location, and the perceived benefits and risks were important factors for participation. Furthermore, suggestions for improving the wetland compensation system and providing alternative livelihoods are proposed to strengthen participation.

The Role of Ect2 Nuclear RhoGEF Activity in Ovarian Cancer Cell Transformation

PubMed Central

Huff, Lauren P.; DeCristo, Molly J.; Trembath, Dimitri; Kuan, Pei Fen; Yim, Margaret; Liu, Jinsong; Cook, Danielle R.; Miller, C. Ryan; Der, Channing J.

2013-01-01

Ect2, a Rho guanine nucleotide exchange factor (RhoGEF), is atypical among RhoGEFs in its predominantly nuclear localization in interphase cells. One current model suggests that Ect2 mislocalization drives cellular transformation by promoting aberrant activation of cytoplasmic Rho family GTPase substrates. However, in ovarian cancers, where Ect2 is both amplified and overexpressed at the mRNA level, we observed that the protein is highly expressed and predominantly nuclear and that nuclear but not cytoplasmic Ect2 increases with advanced disease. Knockdown of Ect2 in ovarian cancer cell lines impaired their anchorage-independent growth without affecting their growth on plastic. Restoration of Ect2 expression rescued the anchorage-independent growth defect, but not if either the DH catalytic domain or the nuclear localization sequences of Ect2 were mutated. These results suggested a novel mechanism whereby Ect2 could drive transformation in ovarian cancer cells by acting as a RhoGEF specifically within the nucleus. Interestingly, Ect2 had an intrinsically distinct GTPase specificity profile in the nucleus versus the cytoplasm. Nuclear Ect2 bound preferentially to Rac1, while cytoplasmic Ect2 bound to RhoA but not Rac. Consistent with nuclear activation of endogenous Rac, Ect2 overexpression was sufficient to recruit Rac effectors to the nucleus, a process that required a functional Ect2 catalytic domain. Furthermore, expression of active nuclearly targeted Rac1 rescued the defect in transformed growth caused by Ect2 knockdown. Our work suggests a novel mechanism of Ect2-driven transformation, identifies subcellular localization as a regulator of GEF specificity, and implicates activation of nuclear Rac1 in cellular transformation. PMID:24386507

A Novel GABRG2 Mutation, p.R136*, in a family with GEFS+ and extended phenotypes

PubMed Central

Shen, Wangzhen; Pickrell, William O.; Cushion, Thomas D.; Davies, Jeffrey S.; Baer, Kristin; Mullins, Jonathan G.L.; Hammond, Carrie L.; Chung, Seo-Kyung; Thomas, Rhys H.; White, Cathy; Smith, Phil E.M.

2014-01-01

Genetic mutations in voltage-gated and ligand-gated ion channel genes have been identified in a small number of Mendelian families with genetic generalised epilepsies (GGEs). They are commonly associated with febrile seizures (FS), childhood absence epilepsy (CAE) and particularly with generalised or genetic epilepsy with febrile seizures plus (GEFS+). In clinical practice, despite efforts to categorise epilepsy and epilepsy families into syndromic diagnoses, many generalised epilepsies remain unclassified with a presumed genetic basis. During the systematic collection of epilepsy families, we assembled a cohort of families with evidence of GEFS+ and screened for variations in the γ2 subunit of the γ-aminobutyric acid (GABA) type A receptor gene (GABRG2). We detected a novel GABRG2(p.R136*) premature translation termination codon in one index-case from a two-generation nuclear family, presenting with an unclassified GGE, a borderline GEFS+ phenotype with learning difficulties and autism spectrum disorder (ASD). The GABRG2(p.R136*) mutation segregates with the febrile seizure component of this family's GGE and is absent in 190 healthy control samples. In vitro expression assays demonstrated that γ2(p.R136*) subunits were produced, but had reduced cell-surface and total expression. When γ2(p.R136*) subunits were co-expressed with α1 and β2 subunits in HEK 293T cells, GABA–evoked currents were reduced. Furthermore, γ2(p.R136*) subunits were highly-expressed in intracellular aggregations surrounding the nucleus and endoplasmic reticulum (ER), suggesting compromised receptor trafficking. A novel GABRG2(p.R136*) mutation extends the spectrum of GABRG2 mutations identified in GEFS+ and GGE phenotypes, causes GABAA receptor dysfunction, and represents a putative epilepsy mechanism. PMID:24407264

A homogeneous quenching resonance energy transfer assay for the kinetic analysis of the GTPase nucleotide exchange reaction.

PubMed

Kopra, Kari; Ligabue, Alessio; Wang, Qi; Syrjänpää, Markku; Blaževitš, Olga; Veltel, Stefan; van Adrichem, Arjan J; Hänninen, Pekka; Abankwa, Daniel; Härmä, Harri

2014-07-01

A quenching resonance energy transfer (QRET) assay for small GTPase nucleotide exchange kinetic monitoring is demonstrated using nanomolar protein concentrations. Small GTPases are central signaling proteins in all eukaryotic cells acting as a "molecular switches" that are active in the GTP-state and inactive in the GDP-state. GTP-loading is highly regulated by guanine nucleotide exchange factors (GEFs). In several diseases, most prominently cancer, this process in misregulated. The kinetics of the nucleotide exchange reaction reports on the enzymatic activity of the GEF reaction system and is, therefore, of special interest. We determined the nucleotide exchange kinetics using europium-labeled GTP (Eu-GTP) in the QRET assay for small GTPases. After GEF catalyzed GTP-loading of a GTPase, a high time-resolved luminescence signal was found to be associated with GTPase bound Eu-GTP, whereas the non-bound Eu-GTP fraction was quenched by soluble quencher. The association kinetics of the Eu-GTP was measured after GEF addition, whereas the dissociation kinetics could be determined after addition of unlabeled GTP. The resulting association and dissociation rates were in agreement with previously published values for H-Ras(Wt), H-Ras(Q61G), and K-Ras(Wt), respectively. The broader applicability of the QRET assay for small GTPases was demonstrated by determining the kinetics of the Ect2 catalyzed RhoA(Wt) GTP-loading. The QRET assay allows the use of nanomolar protein concentrations, as more than 3-fold signal-to-background ratio was achieved with 50 nM GTPase and GEF proteins. Thus, small GTPase exchange kinetics can be efficiently determined in a HTS compatible 384-well plate format.

RhoGTPase Regulators Orchestrate Distinct Stages of Synaptic Development

PubMed Central

Martin-Vilchez, Samuel; Whitmore, Leanna; Asmussen, Hannelore; Zareno, Jessica; Horwitz, Rick; Newell-Litwa, Karen

2017-01-01

Small RhoGTPases regulate changes in post-synaptic spine morphology and density that support learning and memory. They are also major targets of synaptic disorders, including Autism. Here we sought to determine whether upstream RhoGTPase regulators, including GEFs, GAPs, and GDIs, sculpt specific stages of synaptic development. The majority of examined molecules uniquely regulate either early spine precursor formation or later maturation. Specifically, an activator of actin polymerization, the Rac1 GEF β-PIX, drives spine precursor formation, whereas both FRABIN, a Cdc42 GEF, and OLIGOPHRENIN-1, a RhoA GAP, regulate spine precursor elongation. However, in later development, a novel Rac1 GAP, ARHGAP23, and RhoGDIs inactivate actomyosin dynamics to stabilize mature synapses. Our observations demonstrate that specific combinations of RhoGTPase regulatory proteins temporally balance RhoGTPase activity during post-synaptic spine development. PMID:28114311

National Centers for Environmental Prediction

Science.gov Websites

OPERATIONAL 00Z, .... 12Z ... EXPERIMENTAL Daily Comparisons between GFS/GEFS control & ECMWF/ECMWF control 00Z T382/38km GFS, 00Z T190/70km GEFS control 12Z T1279/16km ECMWF, 12Z T639/30km ECMWF ensemble control Daily Values of 500 hPa Height AC, RMS, Talagrand & Outliers Mean of 14 GFS, 10 ECMWF and 16

A non-typical sequence of phase transitions in (NH4)3GeF7: optical and structural characterization.

PubMed

Mel'nikova, S V; Molokeev, M S; Laptash, N M; Misyul, S V

2016-03-28

Single crystals of germanium double salt (NH4)3GeF7 = (NH4)2GeF6·NH4F = (NH4)3[GeF6]F were grown and studied by the methods of polarization optics and X-ray diffraction. The birefringence Δn = (no - ne), the rotation angle of the optical indicatrix ϕ(T) and unit cell parameters were measured in the temperature range 100-400 K. Three structural phase transitions were found at the temperatures: T1↓ = 279.2 K (T1↑ = 279.4 K), T2↑ = 270 K (T2↓ = 268.9 K), T3↓ = 218 K (T3↑ = 227 K). An unusual sequence of symmetry transformations with temperature change was established: P4/mbm (Z = 2) (G1) ↔ Pbam (Z = 4) (G2) ↔ P21/c (Z = 4) (G3) ↔ Pa3[combining macron] (Z = 8) (G4). The crystal structures of different phases were determined. The experimental data were additionally interpreted by a group-theoretical analysis of the complete condensate of order parameters taking into account the critical and noncritical atomic displacements. Strengthening of the N-HF hydrogen bonds can be a driving force of the observed phase transitions.

International Advocacy against DDT and Other Public Health Insecticides for Malaria Control

DTIC Science & Technology

2011-01-19

compared with controls. This was true across all eight countries. Broad use of antimalarial drugs was the primary method of malaria suppression in...eight countries. Broad use of antimalarial drugs was the primary method of malaria suppression in the eight countries, but this method was not a GEF...Available evidence suggests the NMCPs did their work regardless of the presence or absence of GEF project personnel. Thus, antimalarial treatment (the

Trio combines with dock to regulate Pak activity during photoreceptor axon pathfinding in Drosophila.

PubMed

Newsome, T P; Schmidt, S; Dietzl, G; Keleman, K; Asling, B; Debant, A; Dickson, B J

2000-04-28

Correct pathfinding by Drosophila photoreceptor axons requires recruitment of p21-activated kinase (Pak) to the membrane by the SH2-SH3 adaptor Dock. Here, we identify the guanine nucleotide exchange factor (GEF) Trio as another essential component in photoreceptor axon guidance. Regulated exchange activity of one of the two Trio GEF domains is critical for accurate pathfinding. This GEF domain activates Rac, which in turn activates Pak. Mutations in trio result in projection defects similar to those observed in both Pak and dock mutants, and trio interacts genetically with Rac, Pak, and dock. These data define a signaling pathway from Trio to Rac to Pak that links guidance receptors to the growth cone cytoskeleton. We propose that distinct signals transduced via Trio and Dock act combinatorially to activate Pak in spatially restricted domains within the growth cone, thereby controlling the direction of axon extension.

The Resilience Assessment Framework: a common indicator for land management?

NASA Astrophysics Data System (ADS)

Cowie, Annette; Metternicht, Graciela; O'Connell, Deborah

2015-04-01

At the Rio+20 conference in June 2013, the United Nations Convention to Combat Desertification (UNCCD), the Convention on Biological Diversity (CBD), and the United Nations Framework Convention on Climate Change (UNFCCC) reinforced their mutual interests in building linkages between biodiversity conservation, sustainable land management, and climate change mitigation and adaptation. The UNCCD sees building resilience of agro-ecosystems as a common interest that could strengthen linkages between the conventions and deliver synergies in progressing goals of each of the conventions. Furthermore, enhancing resilience of productive agro-ecosystems is fundamental to food security and sustainable development, and thus aligns with the Sustainable Development Goals (SDGs). The Global Environment Facility (GEF) shares the interest of the conventions in building resilience in agro-ecosystems. Indicators of resilience are required for monitoring progress in these endeavors, application of a common indicator between the UNCCD, UNFCCC and CBD as a measure of both land-based adaptation and ecosystem resilience, could strengthen links between the conventions and increase attention to the broad benefits of improved land management. Consequently, the Scientific and Technical Advisory Panel (STAP) to the GEF commissioned the Commonwealth Scientific and Industrial Research Organisation (CSIRO) to produce a report reviewing the conceptual basis for resilience, and proposing an indicator approach that could meet the needs of the Conventions and the GEF for an indicator of agro-ecosystem resilience and land-based adaption. The paper presents a synthesis of scientific understanding of resilience in agro-ecosystems, reviews indicators that have been proposed, and, having concluded that none of the extant indicator approaches adequately assesses resilience of agro-ecosystems, proposes a new approach to the assessment of resilience. Recognizing that no single indicator of resilience is applicable to all agro-ecosystems, and that involvement of stakeholders is critical to discerning the critical variables to be assessed, the proposed framework uses an iterative participatory approach to characterise the system, considering also interactions across and within scales; identify the controlling variables, and assess proximity to thresholds, and adaptive capacity. The framework consists of four elements: Element A: System description; Element B Assessing the system; Element C Adaptive governance and management; Element D Participatory process. Element D is intended as a cross-cutting element, applying across Elements A to C, although Elements A and B can be applied as a desktop activity in a preliminary assessment. The results of the assessment are synthesised in "Resilience action indicators", that summarise the state of the system with respect to the need to adapt or transform. The presentation will summarise the framework and the responses of expert reviewers who identified strengths of the approach, and challenges for implementation, particularly at program and national scales. The presentation will emphasise the conceptual basis for the approach, and the role of scientists in testing, refining and operationalizing the approach.

Improving GEFS Weather Forecasts for Indian Monsoon with Statistical Downscaling

NASA Astrophysics Data System (ADS)

Agrawal, Ankita; Salvi, Kaustubh; Ghosh, Subimal

2014-05-01

Weather forecast has always been a challenging research problem, yet of a paramount importance as it serves the role of 'key input' in formulating modus operandi for immediate future. Short range rainfall forecasts influence a wide range of entities, right from agricultural industry to a common man. Accurate forecasts actually help in minimizing the possible damage by implementing pre-decided plan of action and hence it is necessary to gauge the quality of forecasts which might vary with the complexity of weather state and regional parameters. Indian Summer Monsoon Rainfall (ISMR) is one such perfect arena to check the quality of weather forecast not only because of the level of intricacy in spatial and temporal patterns associated with it, but also the amount of damage it can cause (because of poor forecasts) to the Indian economy by affecting agriculture Industry. The present study is undertaken with the rationales of assessing, the ability of Global Ensemble Forecast System (GEFS) in predicting ISMR over central India and the skill of statistical downscaling technique in adding value to the predictions by taking them closer to evidentiary target dataset. GEFS is a global numerical weather prediction system providing the forecast results of different climate variables at a fine resolution (0.5 degree and 1 degree). GEFS shows good skills in predicting different climatic variables but fails miserably over rainfall predictions for Indian summer monsoon rainfall, which is evident from a very low to negative correlation values between predicted and observed rainfall. Towards the fulfilment of second rationale, the statistical relationship is established between the reasonably well predicted climate variables (GEFS) and observed rainfall. The GEFS predictors are treated with multicollinearity and dimensionality reduction techniques, such as principal component analysis (PCA) and least absolute shrinkage and selection operator (LASSO). Statistical relationship is established between the principal components and observed rainfall over training period and predictions are obtained for testing period. The validations show high improvements in correlation coefficient between observed and predicted data (0.25 to 0.55). The results speak in favour of statistical downscaling methodology which shows the capability to reduce the gap between observed data and predictions. A detailed study is required to be carried out by applying different downscaling techniques to quantify the improvements in predictions.

Improving medium-range and seasonal hydroclimate forecasts in the southeast USA

NASA Astrophysics Data System (ADS)

Tian, Di

Accurate hydro-climate forecasts are important for decision making by water managers, agricultural producers, and other stake holders. Numerical weather prediction models and general circulation models may have potential for improving hydro-climate forecasts at different scales. In this study, forecast analogs of the Global Forecast System (GFS) and Global Ensemble Forecast System (GEFS) based on different approaches were evaluated for medium-range reference evapotranspiration (ETo), irrigation scheduling, and urban water demand forecasts in the southeast United States; the Climate Forecast System version 2 (CFSv2) and the North American national multi-model ensemble (NMME) were statistically downscaled for seasonal forecasts of ETo, precipitation (P) and 2-m temperature (T2M) at the regional level. The GFS mean temperature (Tmean), relative humidity, and wind speed (Wind) reforecasts combined with the climatology of Reanalysis 2 solar radiation (Rs) produced higher skill than using the direct GFS output only. Constructed analogs showed slightly higher skill than natural analogs for deterministic forecasts. Both irrigation scheduling driven by the GEFS-based ETo forecasts and GEFS-based ETo forecast skill were generally positive up to one week throughout the year. The GEFS improved ETo forecast skill compared to the GFS. The GEFS-based analog forecasts for the input variables of an operational urban water demand model were skillful when applied in the Tampa Bay area. The modified operational models driven by GEFS analog forecasts showed higher forecast skill than the operational model based on persistence. The results for CFSv2 seasonal forecasts showed maximum temperature (Tmax) and Rs had the greatest influence on ETo. The downscaled Tmax showed the highest predictability, followed by Tmean, Tmin, Rs, and Wind. The CFSv2 model could better predict ETo in cold seasons during El Nino Southern Oscillation (ENSO) events only when the forecast initial condition was in ENSO. Downscaled P and T2M forecasts were produced by directly downscaling the NMME P and T2M output or indirectly using the NMME forecasts of Nino3.4 sea surface temperatures to predict local-scale P and T2M. The indirect method generally showed the highest forecast skill which occurs in cold seasons. The bias-corrected NMME ensemble forecast skill did not outperform the best single model.

Synthesis and properties of Rb2GeF6:Mn4+ red-emitting phosphors

NASA Astrophysics Data System (ADS)

Sakurai, Shono; Nakamura, Toshihiro; Adachi, Sadao

2018-02-01

Rb2GeF6:Mn4+ red-emitting phosphors were synthesized by coprecipitation and their structural and optical properties were investigated by laser microscopy observation, X-ray diffraction (XRD) analysis, photoluminescence (PL) analysis, PL excitation (PLE) spectroscopy, and PL decay measurement. Single-crystalline ingots in the form of a hexagonal pyramid were prepared with a basal plane diameter of ˜2 mm. The XRD analysis suggested that Rb2GeF6 crystallizes in the hexagonal structure (C6v4 = P63mc) with a = 0.5955 nm and c = 0.9672 nm. The phosphor exhibited the strong Mn4+-related zero-phonon line (ZPL) emission peak typically observed in host crystals with piezoelectrically active lattices such as a hexagonal lattice. The quantum efficiencies of the bulk ingot and powdered samples were 87 and 74%, respectively, with nearly the same luminescence decay time of ˜6 ms. The exact ZPL energies and related crystal-field and Racah parameters were obtained from the PL and PLE spectra by Franck-Condon analysis. Temperature-dependent PL intensities were analyzed from T = 20 to 500 K using a thermal quenching model by considering Bose-Einstein phonon statistics. A comparative discussion on the phosphor properties of Rb2GeF6:Mn4+ and Rb2MF6:Mn4+ with M = Si and Ti was also given.

Brain Region and Isoform-Specific Phosphorylation Alters Kalirin SH2 Domain Interaction Sites and Calpain Sensitivity

PubMed Central

Miller, Megan B.; Yan, Yan; Machida, Kazuya; Kiraly, Drew D.; Levy, Aaron D.; Wu, Yi I.; Lam, TuKiet T.; Abbott, Thomas; Koleske, Anthony J.; Eipper, Betty A.; Mains, Richard E.

2017-01-01

Kalirin7 (Kal7), a postsynaptic Rho GDP/GTP exchange factor (RhoGEF), plays a crucial role in long term potentiation and in the effects of cocaine on behavior and spine morphology. The KALRN gene has been linked to schizophrenia and other disorders of synaptic function. Mass spectrometry was used to quantify phosphorylation at 26 sites in Kal7 from individual adult rat nucleus accumbens and prefrontal cortex before and after exposure to acute or chronic cocaine. Region- and isoform-specific phosphorylation was observed along with region-specific effects of cocaine on Kal7 phosphorylation. Evaluation of the functional significance of multi-site phosphorylation in a complex protein like Kalirin is difficult. With the identification of five tyrosine phosphorylation (pY) sites, a panel of 71 SH2 domains was screened, identifying subsets that interacted with multiple pY sites in Kal7. In addition to this type of reversible interaction, endoproteolytic cleavage by calpain plays an essential role in long-term potentiation. Calpain cleaved Kal7 at two sites, separating the N-terminal domain, which affects spine length, and the PDZ binding motif from the GEF domain. Mutations preventing phosphorylation did not affect calpain sensitivity or GEF activity; phosphomimetic mutations at specific sites altered protein stability, increased calpain sensitivity and reduced GEF activity. PMID:28418645

The Tea4-PP1 landmark promotes local growth by dual Cdc42 GEF recruitment and GAP exclusion.

PubMed

Kokkoris, Kyriakos; Gallo Castro, Daniela; Martin, Sophie G

2014-05-01

Cell polarization relies on small GTPases, such as Cdc42, which can break symmetry through self-organizing principles, and landmarks that define the axis of polarity. In fission yeast, microtubules deliver the Tea1-Tea4 complex to mark cell poles for growth, but how this complex activates Cdc42 is unknown. Here, we show that ectopic targeting of Tea4 to cell sides promotes the local activation of Cdc42 and cell growth. This activity requires that Tea4 binds the type I phosphatase (PP1) catalytic subunit Dis2 or Sds21, and ectopic targeting of either catalytic subunit is similarly instructive for growth. The Cdc42 guanine-nucleotide-exchange factor Gef1 and the GTPase-activating protein Rga4 are required for Tea4-PP1-dependent ectopic growth. Gef1 is recruited to ectopic Tea4 and Dis2 locations to promote Cdc42 activation. By contrast, Rga4 is locally excluded by Tea4, and its forced colocalization with Tea4 blocks ectopic growth, indicating that Rga4 must be present, but at sites distinct from Tea4. Thus, a Tea4-PP1 landmark promotes local Cdc42 activation and growth both through Cdc42 GEF recruitment and by creating a local trough in a Cdc42 GAP.

Coupling of the Matched Gravity and Electromagnetic Fields of the Sun with Jupiter and its Moons Together in Nearest Portion of Jupiter's Orbit to the Sun as the Main Cause of the Peak of Approximately 11 Yearly Solar Cycles and Hazards from Solar Storms

NASA Astrophysics Data System (ADS)

Gholibeigian, Kazem; Gholibeigian, Hassan

2016-04-01

On March 13, 1989 the entire province of Quebec Blackout by solar storm during solar cycle 22. The solar storm of 1859, also known as the Carrington event, was a powerful geomagnetic solar storm during solar cycle 10. The solar storm of 2012 during solar cycle 24 was of similar magnitude, but it passed Earth's orbit without striking the plane. All of these solar storms occurred in the peak of 11 yearly solar cycles. In this way, the White House in its project which is focusing on hazards from solar system, in a new strategy and action plan to increase protection from damaging solar emissions, should focus on coupling of the matched Gravity and Electromagnetic Fields)GEFs) of the Sun with Jupiter and its moons together. On the other hand, in solar system, the Jupiter's gravity has largest effect to the Sun's core and its dislocation, because the gravity force between the Jupiter and the Sun is 11.834 times, In addition overlapping of the solar cycles with the Jupiter's orbit period is 11.856 years. These observable factors lead us to the effect of the Jupiter and Sun gravity fields coupling as the main cause of the approximately 11 years duration for solar cycles. Its peak in each cycle is when the Jupiter is in nearest portion to the Sun in its orbit. In this way, the other planets in their coupling with Sun help to the variations and strengthening solar cycles. [Gholibeigian, 7/24/2015http://adsabs.harvard.edu/abs/2014EGU]. In other words, the both matched GEFs are generating by the large scale forced convection system inside the stars and planets [Gholibeigian et. al, AGU Fall Meeting 2015]. These two fields are couple and strengthening each other. The Jupiter with its 67 moons generate the largest coupled and matched GEFs in its core and consequently strongest effect on the Sun's core. Generation and coupling of the Jupiter's GEFs with its moons like Europa, Io and Ganymede make this planet of thousands of times brighter and many times bigger than Earth as the strongest variable GEFs in solar system after the Sun. For example, Ganymede is the largest moon of Jupiter and in the Solar System. Completing an orbit in roughly seven days. It means that it generates 52 GEFs oscillations (loading, unloading) per year in solar cycle while it is rotating around the Jupiter. New observations of the planet's extreme ultraviolet emissions show that bright explosions of Jupiter's aurora by the planet-moon interaction, not by solar activity [Tomoki Kimura, JAEA]. Coupling of Jupiter's GEFs and its moons with the Sun generate very strong GEFs and approximately 11 yearly solar cycles. The peaks of each cycle is when the Jupiter passes from the nearest portion of its orbit to the Sun. which some of its peaks generate gigantic solar storms and hazards to the Earth. The Earth passes from between of Sun and Jupiter eleven times in each solar cycle and may be under shooting of storms from the both side specially during 2-3 years in cycle's peak.

Synergistic antitumor effect between gefitinib and fractionated irradiation in anaplastic oligodendrogliomas cannot be predicted by the Egfr signaling activity.

PubMed

Pinel, Sophie; Mriouah, Jihane; Vandamme, Marc; Chateau, Alicia; Plénat, François; Guérin, Eric; Taillandier, Luc; Bernier-Chastagner, Valérie; Merlin, Jean-Louis; Chastagner, Pascal

2013-01-01

In high-grade gliomas, the identification of patients that could benefit from EGFR inhibitors remains a challenge, hindering the use of these agents. Using xenografts models, we evaluated the antitumor effect of the combined treatment "gefitinib + radiotherapy" and aimed to identify the profile of responsive tumors. Expression of phosphorylated proteins involved in the EGFR-dependent signaling pathways was analyzed in 10 glioma models. We focused on three models of anaplastic oligodendrogliomas (TCG2, TCG3 and TCG4) harboring high levels of phospho-EGFR, phospho-AKT and phospho-MEK1. They were treated with gefitinib (GEF 75 mg/kg/day x 5 days/week, for 2 weeks) and/or fractionated radiotherapy (RT: 5x2Gy/week for 2 weeks). Our results showed that GEF and/or RT induced significant tumor growth delays. However, only the TCG3 xenografts were highly responsive to the combination GEF+RT, with ∼50% of tumor cure. Phosphoproteins analysis five days after treatment onset demonstrated in TCG3 xenografts, but not in TCG2 model, that the EGFR-dependent pathways were inhibited after GEF treatment. Moreover, TCG3-bearing mice receiving GEF monotherapy exhibited a transient beneficial therapeutic response, rapidly followed by tumor regrowth, along with a major vascular remodeling. Taken together, our data evoked an "EGFR-addictive" behavior for TCG3 tumors. This study confirms that combination of gefitinib with fractionated irradiation could be a potent therapeutic strategy for anaplastic oligodendrogliomas harboring EGFR abnormalities but this treatment seems mainly beneficial for "EGFR-addictive" tumors. Unfortunately, neither the usual molecular markers (EGFR amplification, PTEN loss) nor the basal overexpression of phosphoproteins were useful to distinguish this responsive tumor. Evaluating the impact of TKIs on the EGFR-dependent pathways during the treatment might be more relevant, and requires further validation.

TD-60 links RalA GTPase function to the CPC in mitosis

PubMed Central

Papini, Diana; Langemeyer, Lars; Abad, Maria A.; Kerr, Alastair; Samejima, Itaru; Eyers, Patrick A.; Jeyaprakash, A. Arockia; Higgins, Jonathan M. G.; Barr, Francis A.; Earnshaw, William C.

2015-01-01

TD-60 (also known as RCC2) is a highly conserved protein that structurally resembles the Ran guanine exchange factor (GEF) RCC1, but has not previously been shown to have GEF activity. TD-60 has a typical chromosomal passenger complex (CPC) distribution in mitotic cells, but associates with integrin complexes and is involved in cell motility during interphase. Here we show that TD-60 exhibits GEF activity, in vitro and in cells, for the small GTPase RalA. TD-60 or RalA depletion causes spindle abnormalities in prometaphase associated with abnormal centromeric accumulation of CPC components. TD-60 and RalA apparently work together to contribute to the regulation of kinetochore–microtubule interactions in early mitosis. Importantly, several mitotic phenotypes caused by TD-60 depletion are reverted by the expression of a GTP-locked mutant, RalA (Q72L). The demonstration that a small GTPase participates in the regulation of the CPC reveals a level of mitotic regulation not suspected in previous studies. PMID:26158537

Epidermal wound repair is regulated by the planar cell polarity signaling pathway.

PubMed

Caddy, Jacinta; Wilanowski, Tomasz; Darido, Charbel; Dworkin, Sebastian; Ting, Stephen B; Zhao, Quan; Rank, Gerhard; Auden, Alana; Srivastava, Seema; Papenfuss, Tony A; Murdoch, Jennifer N; Humbert, Patrick O; Parekh, Vishwas; Boulos, Nidal; Weber, Thomas; Zuo, Jian; Cunningham, John M; Jane, Stephen M

2010-07-20

The mammalian PCP pathway regulates diverse developmental processes requiring coordinated cellular movement, including neural tube closure and cochlear stereociliary orientation. Here, we show that epidermal wound repair is regulated by PCP signaling. Mice carrying mutant alleles of PCP genes Vangl2, Celsr1, PTK7, and Scrb1, and the transcription factor Grhl3, interact genetically, exhibiting failed wound healing, neural tube defects, and disordered cochlear polarity. Using phylogenetic analysis, ChIP, and gene expression in Grhl3(-)(/-) mice, we identified RhoGEF19, a homolog of a RhoA activator involved in PCP signaling in Xenopus, as a direct target of GRHL3. Knockdown of Grhl3 or RhoGEF19 in keratinocytes induced defects in actin polymerization, cellular polarity, and wound healing, and re-expression of RhoGEF19 rescued these defects in Grhl3-kd cells. These results define a role for Grhl3 in PCP signaling and broadly implicate this pathway in epidermal repair. (c) 2010 Elsevier Inc. All rights reserved.

Epidermal wound repair is regulated by the planar cell polarity signaling pathway

PubMed Central

Caddy, Jacinta; Wilanowski, Tomasz; Darido, Charbel; Dworkin, Sebastian; Ting, Stephen B.; Zhao, Quan; Rank, Gerhard; Auden, Alana; Srivastava, Seema; Papenfuss, Tony A.; Murdoch, Jennifer N.; Humbert, Patrick O.; Boulos, Nidal; Weber, Thomas; Zuo, Jian; Cunningham, John M.; Jane, Stephen M.

2010-01-01

SUMMARY The mammalian PCP pathway regulates diverse developmental processes requiring coordinated cellular movement, including neural tube closure and cochlear stereociliary orientation. Here, we show that epidermal wound repair is regulated by PCP signaling. Mice carrying mutant alleles of PCP genes Vangl2, Celsr1, PTK7, and Scrb1, and the transcription factor Grhl3, interact genetically, exhibiting failed wound healing, neural tube defects and disordered cochlear polarity. Using phylogenetic analysis, ChIP, and gene expression in Grhl3−/− mice, we identified RhoGEF19, a homologue of a RhoA activator involved in PCP signaling in Xenopus, as a direct target of GRHL3. Knockdown of Grhl3 or RhoGEF19 in keratinocytes induced defects in actin polymerisation, cellular polarity and wound healing, and re-expression of RhoGEF19 rescued these defects in Grhl3-kd cells. These results define a role for Grhl3 in PCP signaling, and broadly implicate this pathway in epidermal repair. PMID:20643356

[Exon-dependent Subgroup-analysis of the Non-interventional REASON-Study: PFS and OS in EGFR-mutated NSCLC Patients Treated with Gefitinib or Chemotherapy].

PubMed

Schuette, W; Dietel, M; Thomas, M; Eberhardt, W; Griesinger, F; Zirrgiebel, U; Radke, S; Schirmacher, P

2016-08-01

To analyze the influence of the localization of mutations in the epidermal growth factor receptor (EGFR) gene on progression-free (PFS) and overall survival (OS) in patients (pts) with locally advanced or metastatic non-small cell lung cancer (NSCLC) treated with gefitinib (gef) or chemotherapy (CT) under real world conditions within the REASON study. Subgroups of pts with mutations in exon 19 (n = 141), 18/20 (n = 43), and 21 (n = 104) were analyzed for PFS and OS according to gef or CT treatment and compared using the log-rank test. Pts with mutations in exon 19 and 18/20 treated with gef as first line therapy showed increased PFS and OS compared to CT. This increase was statistically significant in pts with exon 19 mutation (11.3 vs. 6.5 months), but was not found in pts with exon 21 mutation (9.1 vs. 9.3 months). Also, OS was significantly increased in patients with mutation in exon 19 treated with gef ever over all treatment lines compared to CT (21.8 vs. 10.6 months), whereas this was not found in pts with mutation in exon 21 (14.1 vs. 13.9 months). Localization and nature of EGFR mutations influences gefitinib treatment outcomes under routine conditions and should therefore be analyzed in detail. © Georg Thieme Verlag KG Stuttgart · New York.

Virtual photon emission from a quark-gluon plasma

DOE Office of Scientific and Technical Information (OSTI.GOV)

Suryanarayana, S. V.

We present phenomenological formulas for virtual photon emission rates from a thermalized quark-gluon plasma (QGP) that include bremsstrahlung and annihilation with scattering (AWS) mechanisms along with the Landau-Pomeranchuk-Migdal (LPM) effects. For this purpose we follow the approach of generalized emission functions (GEF) for virtual photon emission, we showed earlier for a fixed temperature and strong coupling constant. In the present work, we extend the LPM calculations for several temperatures and strong coupling strengths, photon energies (q{sub 0}), photon mass (Q{sup 2}), and quark energies (p{sub 0}). We generalize the dynamical scaling variables, x{sub T},x{sub L}, for bremsstrahlung and AWS processesmore » that are now functions of variables p{sub 0},q{sub 0},Q{sup 2},T,{alpha}{sub s}. The GEF introduced earlier, g{sub T}{sup b},g{sub T}{sup a},g{sub L}{sup b},g{sub L}{sup a}, are also generalized for any temperatures and coupling strengths. From this, the imaginary part of the photon polarization tensor as a function of photon mass and energy has been calculated as a one-dimensional integral over these GEF and parton distribution functions in the plasma. By fitting these polarization tensors obtained from GEF method, we obtained a phenomenological formula for virtual photon emission rates as a function of (q{sub 0},Q{sup 2},T,{alpha}{sub s}) that includes bremsstrahlung and AWS mechanisms with LPM effects.« less

Far-side geometrical enhancement in surface-enhanced Raman scattering with Ag plasmonic films

NASA Astrophysics Data System (ADS)

Perera, M. Nilusha M. N.; Gibbs, W. E. Keith; Juodkazis, Saulius; Stoddart, Paul R.

2018-01-01

Surface-enhanced Raman scattering (SERS) is a surface sensitive technique where the large increase in scattering has primarily been attributed to electromagnetic and chemical enhancements. While smaller geometrical enhancements due to thin film interference and cavity resonances have also been reported, an additional enhancement in the SERS signal, referred to as the `far-side geometrical enhancement', occurs when the SERS substrate is excited through an underlying transparent dielectric substrate. Here the far-side geometrically-enhanced SERS signal has been explored experimentally in more detail. Thermally evaporated Ag plasmonic films functionalised with thiophenol were used to study the dependence of the geometrically-enhanced SERS signal on the excitation wavelength, supporting substrate material and excitation angle of incidence. The results were interpreted using a `geometrical enhancement factor' (GEF), defined as the ratio of far-side to near-side SERS signal intensity. The experimental results confirmed that the highest GEFs of 3.2-3.5× are seen closer to the localized surface plasmon resonance peak of the Ag metallic nanostructures. Interestingly, the GEF for Ag plasmonic films deposited on glass and sapphire were the same within the measurement errors, whereas increasing angle of incidence showed a decrease in the GEF. Given this improved understanding of the far-side geometrical SERS enhancement, the potential for further signal amplification and optimisation for practical sensing applications can now be considered, especially for SERS detection modes at the farend of optical fibre probes and through process windows.

The Aarskog-Scott Syndrome Protein Fgd1 Regulates Podosome Formation and Extracellular Matrix Remodeling in Transforming Growth Factor β-Stimulated Aortic Endothelial Cells ▿

PubMed Central

Daubon, Thomas; Buccione, Roberto; Génot, Elisabeth

2011-01-01

Podosomes are dynamic actin-rich adhesion plasma membrane microdomains endowed with extracellular matrix-degrading activities. In aortic endothelial cells, podosomes are induced by transforming growth factor β (TGF-β), but how this occurs is largely unknown. It is thought that, in endothelial cells, podosomes play a role in vessel remodeling and/or in breaching anatomical barriers. We demonstrate here that, in bovine aortic endothelial cells, that the Cdc42-specific guanine exchange factor (GEF) Fgd1 is expressed and regulated by TGF-β to induce Cdc42-dependent podosome assembly. Within 15 min of TGF-β stimulation, Fgd1, but none of the other tested Cdc42 GEFs, undergoes tyrosine phosphorylation, associates with Cdc42, and translocates to the subcortical cytoskeleton via a cortactin-dependent mechanism. Small interfering RNA-mediated Fgd1 knockdown inhibits TGF-β-induced Cdc42 activation. Fgd1 depletion also reduces podosome formation and associated matrix degradation and these defects are rescued by reexpression of Fgd1. Although overexpression of Fgd1 does not promote podosome formation per se, it enhances TGF-β-induced matrix degradation. Our results identify Fgd1 as a TGF-β-regulated GEF and, as such, the first GEF to be involved in the process of cytokine-induced podosome formation. Our findings reveal the involvement of Fgd1 in endothelial cell biology and open up new avenues to study its role in vascular pathophysiology. PMID:21911474

Evidence for the Involvement of Lfc and Tctex-1 in Axon Formation

PubMed Central

Conde, Cecilia; Arias, Cristina; Robin, Maria; Li, Aiqun; Saito, Masaki; Chuang, Jen-Zen; Nairn, Angus C.; Sung, Ching-Hwa; Cáceres, Alfredo

2013-01-01

RhoA and Rac play key and opposite roles during neuronal polarization. We now show that Lfc, a guanosine nucleotide exchange factor (GEF), localizes to the Golgi apparatus and growth cones of developing neurons and negatively regulates neurite sprouting and axon formation through a Rho signaling pathway. Tctex-1, a dynein light chain implicated in axon outgrowth by modulating actin dynamics and Rac activity, colocalizes and physically interacts with Lfc, thus inhibiting its GEF activity, decreasing Rho-GTP levels, and functionally antagonizing Lfc during neurite formation. PMID:20463241

Which Way In? The RalF Arf-GEF Orchestrates Rickettsia Host Cell Invasion

PubMed Central

Rennoll-Bankert, Kristen E.; Rahman, M. Sayeedur; Gillespie, Joseph J.; Guillotte, Mark L.; Kaur, Simran J.; Lehman, Stephanie S.; Beier-Sexton, Magda; Azad, Abdu F.

2015-01-01

Bacterial Sec7-domain-containing proteins (RalF) are known only from species of Legionella and Rickettsia, which have facultative and obligate intracellular lifestyles, respectively. L. pneumophila RalF, a type IV secretion system (T4SS) effector, is a guanine nucleotide exchange factor (GEF) of ADP-ribosylation factors (Arfs), activating and recruiting host Arf1 to the Legionella-containing vacuole. In contrast, previous in vitro studies showed R. prowazekii (Typhus Group) RalF is a functional Arf-GEF that localizes to the host plasma membrane and interacts with the actin cytoskeleton via a unique C-terminal domain. As RalF is differentially encoded across Rickettsia species (e.g., pseudogenized in all Spotted Fever Group species), it may function in lineage-specific biology and pathogenicity. Herein, we demonstrate RalF of R. typhi (Typhus Group) interacts with the Rickettsia T4SS coupling protein (RvhD4) via its proximal C-terminal sequence. RalF is expressed early during infection, with its inactivation via antibody blocking significantly reducing R. typhi host cell invasion. For R. typhi and R. felis (Transitional Group), RalF ectopic expression revealed subcellular localization with the host plasma membrane and actin cytoskeleton. Remarkably, R. bellii (Ancestral Group) RalF showed perinuclear localization reminiscent of ectopically expressed Legionella RalF, for which it shares several structural features. For R. typhi, RalF co-localization with Arf6 and PI(4,5)P2 at entry foci on the host plasma membrane was determined to be critical for invasion. Thus, we propose recruitment of PI(4,5)P2 at entry foci, mediated by RalF activation of Arf6, initiates actin remodeling and ultimately facilitates bacterial invasion. Collectively, our characterization of RalF as an invasin suggests that, despite carrying a similar Arf-GEF unknown from other bacteria, different intracellular lifestyles across Rickettsia and Legionella species have driven divergent roles for RalF during infection. Furthermore, our identification of lineage-specific Arf-GEF utilization across some rickettsial species illustrates different pathogenicity factors that define diverse agents of rickettsial diseases. PMID:26291822

Dissociation of Rac1(GDP)·RhoGDI Complexes by the Cooperative Action of Anionic Liposomes Containing Phosphatidylinositol 3,4,5-Trisphosphate, Rac Guanine Nucleotide Exchange Factor, and GTP*

PubMed Central

Ugolev, Yelena; Berdichevsky, Yevgeny; Weinbaum, Carolyn; Pick, Edgar

2008-01-01

Rac plays a pivotal role in the assembly of the superoxide-generating NADPH oxidase of phagocytes. In resting cells, Rac is found in the cytosol in complex with Rho GDP dissociation inhibitor (RhoGDI). NADPH oxidase assembly involves dissociation of the Rac·RhoGDI complex and translocation of Rac to the membrane. We reported that liposomes containing high concentrations of monovalent anionic phospholipids cause Rac·RhoGDI complex dissociation (Ugolev, Y., Molshanski-Mor, S., Weinbaum, C., and Pick, E. (2006) J. Biol. Chem.281 ,19204 -1921916702219). We now designed an in vitro model mimicking membrane phospholipid remodeling during phagocyte stimulation in vivo. We showed that liposomes of “resting cell membrane” composition (less than 20 mol % monovalent anionic phospholipids), supplemented with 1 mol % of polyvalent anionic phosphatidylinositol 3,4,5-trisphosphate (PtdIns(3,4,5)P3) in conjunction with constitutively active forms of the guanine nucleotide exchange factors (GEFs) for Rac, Trio, or Tiam1 and a non-hydrolyzable GTP analogue, cause dissociation of Rac1(GDP)·RhoGDI complexes, GDP to GTP exchange on Rac1, and binding of Rac1(GTP) to the liposomes. Complexes were not dissociated in the absence of GEF and GTP, and optimal dissociation required the presence of PtdIns(3,4,5)P3 in the liposomes. Dissociation of Rac1(GDP)·RhoGDI complexes was correlated with the affinity of particular GEF constructs, via the N-terminal pleckstrin homology domain, for PtdIns(3,4,5)P3 and involved GEF-mediated GDP to GTP exchange on Rac1. Phagocyte membranes enriched in PtdIns(3,4,5)P3 responded by NADPH oxidase activation upon exposure in vitro to Rac1(GDP)·RhoGDI complexes, p67phox, GTP, and Rac GEF constructs with affinity for PtdIns(3,4,5)P3 at a level superior to that of native membranes. PMID:18505730

The Evolutionary Landscape of Dbl-Like RhoGEF Families: Adapting Eukaryotic Cells to Environmental Signals

PubMed Central

Blangy, Anne

2017-01-01

Abstract The dynamics of cell morphology in eukaryotes is largely controlled by small GTPases of the Rho family. Rho GTPases are activated by guanine nucleotide exchange factors (RhoGEFs), of which diffuse B-cell lymphoma (Dbl)-like members form the largest family. Here, we surveyed Dbl-like sequences from 175 eukaryotic genomes and illuminate how the Dbl family evolved in all eukaryotic supergroups. By combining probabilistic phylogenetic approaches and functional domain analysis, we show that the human Dbl-like family is made of 71 members, structured into 20 subfamilies. The 71 members were already present in ancestral jawed vertebrates, but several members were subsequently lost in specific clades, up to 12% in birds. The jawed vertebrate repertoire was established from two rounds of duplications that occurred between tunicates, cyclostomes, and jawed vertebrates. Duplicated members showed distinct tissue distributions, conserved at least in Amniotes. All 20 subfamilies have members in Deuterostomes and Protostomes. Nineteen subfamilies are present in Porifera, the first phylum that diverged in Metazoa, 14 in Choanoflagellida and Filasterea, single-celled organisms closely related to Metazoa and three in Fungi, the sister clade to Metazoa. Other eukaryotic supergroups show an extraordinary variability of Dbl-like repertoires as a result of repeated and independent gain and loss events. Last, we observed that in Metazoa, the number of Dbl-like RhoGEFs varies in proportion of cell signaling complexity. Overall, our analysis supports the conclusion that Dbl-like RhoGEFs were present at the origin of eukaryotes and evolved as highly adaptive cell signaling mediators. PMID:28541439

A BAC transgenic mouse model reveals neuron subtype-specific effects of a Generalized Epilepsy with Febrile Seizures Plus (GEFS+) mutation

PubMed Central

Tang, Bin; Dutt, Karoni; Papale, Ligia; Rusconi, Raffaella; Shankar, Anupama; Hunter, Jessica; Tufik, Sergio; Yu, Frank H.; Catterall, William A.; Mantegazza, Massimo; Goldin, Alan L.; Escayg, Andrew

2009-01-01

Mutations in the voltage-gated sodium channel SCN1A are responsible for a number of seizure disorders including Generalized Epilepsy with Febrile Seizures Plus (GEFS+) and Severe Myoclonic Epilepsy of Infancy (SMEI). To determine the effects of SCN1A mutations on channel function in vivo, we generated a bacterial artificial chromosome (BAC) transgenic mouse model that expresses the human SCN1A GEFS+ mutation, R1648H. Mice with the R1648H mutation exhibit a more severe response to the proconvulsant kainic acid compared with mice expressing a control Scn1a transgene. Electrophysiological analysis of dissociated neurons from mice with the R1648H mutation reveal delayed recovery from inactivation and increased use-dependent inactivation only in inhibitory bipolar neurons, as well as a hyperpolarizing shift in the voltage dependence of inactivation only in excitatory pyramidal neurons. These results demonstrate that the effects of SCN1A mutations are cell type-dependent and that the R1648H mutation specifically leads to a reduction in interneuron excitability. PMID:19409490

TGF-β regulates LARG and GEF-H1 during EMT to affect stiffening response to force and cell invasion

PubMed Central

Osborne, Lukas D.; Li, George Z.; How, Tam; O'Brien, E. Tim; Blobe, Gerard C.; Superfine, Richard; Mythreye, Karthikeyan

2014-01-01

Recent studies implicate a role for cell mechanics in cancer progression. The epithelial-to-mesenchymal transition (EMT) regulates the detachment of cancer cells from the epithelium and facilitates their invasion into stromal tissue. Although classic EMT hallmarks include loss of cell–cell adhesions, morphology changes, and increased invasion capacity, little is known about the associated mechanical changes. Previously, force application on integrins has been shown to initiate cytoskeletal rearrangements that result in increased cell stiffness and a stiffening response. Here we demonstrate that transforming growth factor β (TGF-β)–induced EMT results in decreased stiffness and loss of the normal stiffening response to force applied on integrins. We find that suppression of the RhoA guanine nucleotide exchange factors (GEFs) LARG and GEF-H1 through TGF-β/ALK5–enhanced proteasomal degradation mediates these changes in cell mechanics and affects EMT-associated invasion. Taken together, our results reveal a functional connection between attenuated stiffness and stiffening response and the increased invasion capacity acquired after TGF-β–induced EMT. PMID:25143398

In vitro guanine nucleotide exchange activity of DHR-2/DOCKER/CZH2 domains.

PubMed

Côté, Jean-François; Vuori, Kristiina

2006-01-01

Rho family GTPases regulate a large variety of biological processes, including the reorganization of the actin cytoskeleton. Like other members of the Ras superfamily of small GTP-binding proteins, Rho GTPases cycle between a GDP-bound (inactive) and a GTP-bound (active) state, and, when active, the GTPases relay extracellular signals to a large number of downstream effectors. Guanine nucleotide exchange factors (GEFs) promote the exchange of GDP for GTP on Rho GTPases, thereby activating them. Most Rho-GEFs mediate their effects through their signature domain known as the Dbl Homology-Pleckstrin Homology (DH-PH) module. Recently, we and others identified a family of evolutionarily conserved, DOCK180-related proteins that also display GEF activity toward Rho GTPases. The DOCK180-family of proteins lacks the canonical DH-PH module. Instead, they rely on a novel domain, termed DHR-2, DOCKER, or CZH2, to exchange GDP for GTP on Rho targets. In this chapter, the experimental approach that we used to uncover the exchange activity of the DHR-2 domain of DOCK180-related proteins will be described.

The tight-adhesion proteins TadGEF of Bradyrhizobium diazoefficiens USDA 110 are involved in cell adhesion and infectivity on soybean roots.

PubMed

Mongiardini, Elías J; Parisi, Gustavo D; Quelas, Juan I; Lodeiro, Aníbal R

2016-01-01

Adhesion of symbiotic bacteria to host plants is an essential early step of the infection process that leads to the beneficial interaction. In the Bradyrhizobium diazoefficiens-soybean symbiosis few molecular determinants of adhesion are known. Here we identified the tight-adhesion gene products TadGEF in the open-reading frames blr3941-blr3943 of the B. diazoefficiens USDA 110 complete genomic sequence. Predicted structure of TadG indicates a transmembrane domain and two extracytosolic domains, from which the C-terminal has an integrin fold. TadE and TadF are also predicted as bearing transmembrane segments. Mutants in tadG or the small cluster tadGEF were impaired in adhesion to soybean roots, and the root infection was delayed. However, nodule histology was not compromised by the mutations, indicating that these effects were restricted to the earliest contact of the B. diazoefficiens and root surfaces. Knowledge of preinfection determinants is important for development of inoculants that are applied to soybean crops worldwide. Copyright © 2015 Elsevier GmbH. All rights reserved.

GDP-bound and nucleotide-free intermediates of the guanine nucleotide exchange in the Rab5·Vps9 system.

PubMed

Uejima, Tamami; Ihara, Kentaro; Goh, Tatsuaki; Ito, Emi; Sunada, Mariko; Ueda, Takashi; Nakano, Akihiko; Wakatsuki, Soichi

2010-11-19

Many GTPases regulate intracellular transport and signaling in eukaryotes. Guanine nucleotide exchange factors (GEFs) activate GTPases by catalyzing the exchange of their GDP for GTP. Here we present crystallographic and biochemical studies of a GEF reaction with four crystal structures of Arabidopsis thaliana ARA7, a plant homolog of Rab5 GTPase, in complex with its GEF, VPS9a, in the nucleotide-free and GDP-bound forms, as well as a complex with aminophosphonic acid-guanylate ester and ARA7·VPS9a(D185N) with GDP. Upon complex formation with ARA7, VPS9 wedges into the interswitch region of ARA7, inhibiting the coordination of Mg(2+) and decreasing the stability of GDP binding. The aspartate finger of VPS9a recognizes GDP β-phosphate directly and pulls the P-loop lysine of ARA7 away from GDP β-phosphate toward switch II to further destabilize GDP for its release during the transition from the GDP-bound to nucleotide-free intermediates in the nucleotide exchange reaction.

Baseline predictability of daily east Asian summer monsoon circulation indices

NASA Astrophysics Data System (ADS)

Ai, Shucong; Chen, Quanliang; Li, Jianping; Ding, Ruiqiang; Zhong, Quanjia

2017-05-01

The nonlinear local Lyapunov exponent (NLLE) method is adopted to quantitatively determine the predictability limit of East Asian summer monsoon (EASM) intensity indices on a synoptic timescale. The predictability limit of EASM indices varies widely according to the definitions of indices. EASM indices defined by zonal shear have a limit of around 7 days, which is higher than the predictability limit of EASM indices defined by sea level pressure (SLP) difference and meridional wind shear (about 5 days). The initial error of EASM indices defined by SLP difference and meridional wind shear shows a faster growth than indices defined by zonal wind shear. Furthermore, the indices defined by zonal wind shear appear to fluctuate at lower frequencies, whereas the indices defined by SLP difference and meridional wind shear generally fluctuate at higher frequencies. This result may explain why the daily variability of the EASM indices defined by zonal wind shear tends be more predictable than those defined by SLP difference and meridional wind shear. Analysis of the temporal correlation coefficient (TCC) skill for EASM indices obtained from observations and from NCEP's Global Ensemble Forecasting System (GEFS) historical weather forecast dataset shows that GEFS has a higher forecast skill for the EASM indices defined by zonal wind shear than for indices defined by SLP difference and meridional wind shear. The predictability limit estimated by the NLLE method is shorter than that in GEFS. In addition, the June-September average TCC skill for different daily EASM indices shows significant interannual variations from 1985 to 2015 in GEFS. However, the TCC for different types of EASM indices does not show coherent interannual fluctuations.

Mechanisms of CDC-42 activation during contact-induced cell polarization.

PubMed

Chan, Emily; Nance, Jeremy

2013-04-01

Polarization of early embryos provides a foundation to execute essential patterning and morphogenetic events. In Caenorhabditis elegans, cell contacts polarize early embryos along their radial axis by excluding the cortical polarity protein PAR-6 from sites of cell contact, thereby restricting PAR-6 to contact-free cell surfaces. Radial polarization requires the cortically enriched Rho GTPase CDC-42, which in its active form recruits PAR-6 through direct binding. The Rho GTPase activating protein (RhoGAP) PAC-1, which localizes specifically to cell contacts, triggers radial polarization by inactivating CDC-42 at these sites. The mechanisms responsible for activating CDC-42 at contact-free surfaces are unknown. Here, in an overexpression screen of Rho guanine nucleotide exchange factors (RhoGEFs), which can activate Rho GTPases, we identify CGEF-1 and ECT-2 as RhoGEFs that act through CDC-42 to recruit PAR-6 to the cortex. We show that ECT-2 and CGEF-1 localize to the cell surface and that removing their activity causes a reduction in levels of cortical PAR-6. Through a structure-function analysis, we show that the tandem DH-PH domains of CGEF-1 and ECT-2 are sufficient for GEF activity, but that regions outside of these domains target each protein to the cell surface. Finally, we provide evidence suggesting that the N-terminal region of ECT-2 may direct its in vivo preference for CDC-42 over another known target, the Rho GTPase RHO-1. We propose that radial polarization results from a competition between RhoGEFs, which activate CDC-42 throughout the cortex, and the RhoGAP PAC-1, which inactivates CDC-42 at cell contacts.

Comparative studies on the human serum albumin binding of the clinically approved EGFR inhibitors gefitinib, erlotinib, afatinib, osimertinib and the investigational inhibitor KP2187.

PubMed

Dömötör, Orsolya; Pelivan, Karla; Borics, Attila; Keppler, Bernhard K; Kowol, Christian R; Enyedy, Éva A

2018-05-30

Binding interactions between human serum albumin (HSA) and four approved epidermal growth factor receptor (EGFR) inhibitors gefitinib (GEF), erlotinib (ERL), afatinib (AFA), osimertinib (OSI), as well as the experimental drug KP2187, were investigated by means of spectrofluorometric and molecular modelling methods. Steady-state and time resolved spectrofluorometric techniques were carried out, including direct quenching of protein fluorescence and site marker displacement measurements. Proton dissociation processes and solvent dependent fluorescence properties were investigated as well. The EGFR inhibitors were predominantly presented in their single protonated form (HL + ) at physiological pH except ERL, which is charge-neutral. Significant solvent dependent fluorescence properties were found for GEF, ERL and KP2187, namely their emission spectra show strong dependence on the polarity and the hydrogen bonding ability of the solvents. The inhibitors proved to be bound at site I of HSA (in subdomain IIA) in a weak-to-moderate fashion (logK' 3.9-4.9) using spectrofluorometry. OSI (logK' 4.3) and KP2187 can additionally bind in site II (in subdomain IIIA), while GEF, ERL and AFA clearly show no interaction here. Docking methods qualitatively confirmed binding site preferences of compounds GEF and KP2187, and indicated that they probably bind to HSA in their neutral forms. Binding constants calculated on the basis of the various experimental data indicate a weak-to-moderate binding on HSA, only OSI exhibits somewhat higher affinity towards this protein. However, model calculations performed at physiological blood concentrations of HSA resulted in high (ca. 90%) bound fractions for the inhibitors, highlighting the importance of plasma protein binding. Copyright © 2018 Elsevier B.V. All rights reserved.

Mechanisms of CDC-42 activation during contact-induced cell polarization

PubMed Central

Chan, Emily; Nance, Jeremy

2013-01-01

Summary Polarization of early embryos provides a foundation to execute essential patterning and morphogenetic events. In Caenorhabditis elegans, cell contacts polarize early embryos along their radial axis by excluding the cortical polarity protein PAR-6 from sites of cell contact, thereby restricting PAR-6 to contact-free cell surfaces. Radial polarization requires the cortically enriched Rho GTPase CDC-42, which in its active form recruits PAR-6 through direct binding. The Rho GTPase activating protein (RhoGAP) PAC-1, which localizes specifically to cell contacts, triggers radial polarization by inactivating CDC-42 at these sites. The mechanisms responsible for activating CDC-42 at contact-free surfaces are unknown. Here, in an overexpression screen of Rho guanine nucleotide exchange factors (RhoGEFs), which can activate Rho GTPases, we identify CGEF-1 and ECT-2 as RhoGEFs that act through CDC-42 to recruit PAR-6 to the cortex. We show that ECT-2 and CGEF-1 localize to the cell surface and that removing their activity causes a reduction in levels of cortical PAR-6. Through a structure–function analysis, we show that the tandem DH-PH domains of CGEF-1 and ECT-2 are sufficient for GEF activity, but that regions outside of these domains target each protein to the cell surface. Finally, we provide evidence suggesting that the N-terminal region of ECT-2 may direct its in vivo preference for CDC-42 over another known target, the Rho GTPase RHO-1. We propose that radial polarization results from a competition between RhoGEFs, which activate CDC-42 throughout the cortex, and the RhoGAP PAC-1, which inactivates CDC-42 at cell contacts. PMID:23424200

GIV/Girdin activates Gαi and inhibits Gαs via the same motif

PubMed Central

Gupta, Vijay; Bhandari, Deepali; Leyme, Anthony; Aznar, Nicolas; Midde, Krishna K.; Lo, I-Chung; Ear, Jason; Niesman, Ingrid; López-Sánchez, Inmaculada; Blanco-Canosa, Juan Bautista; von Zastrow, Mark; Garcia-Marcos, Mikel; Farquhar, Marilyn G.; Ghosh, Pradipta

2016-01-01

We previously showed that guanine nucleotide-binding (G) protein α subunit (Gα)-interacting vesicle-associated protein (GIV), a guanine-nucleotide exchange factor (GEF), transactivates Gα activity-inhibiting polypeptide 1 (Gαi) proteins in response to growth factors, such as EGF, using a short C-terminal motif. Subsequent work demonstrated that GIV also binds Gαs and that inactive Gαs promotes maturation of endosomes and shuts down mitogenic MAPK–ERK1/2 signals from endosomes. However, the mechanism and consequences of dual coupling of GIV to two G proteins, Gαi and Gαs, remained unknown. Here we report that GIV is a bifunctional modulator of G proteins; it serves as a guanine nucleotide dissociation inhibitor (GDI) for Gαs using the same motif that allows it to serve as a GEF for Gαi. Upon EGF stimulation, GIV modulates Gαi and Gαs sequentially: first, a key phosphomodification favors the assembly of GIV–Gαi complexes and activates GIV’s GEF function; then a second phosphomodification terminates GIV’s GEF function, triggers the assembly of GIV–Gαs complexes, and activates GIV’s GDI function. By comparing WT and GIV mutants, we demonstrate that GIV inhibits Gαs activity in cells responding to EGF. Consequently, the cAMP→PKA→cAMP response element-binding protein signaling axis is inhibited, the transit time of EGF receptor through early endosomes are accelerated, mitogenic MAPK–ERK1/2 signals are rapidly terminated, and proliferation is suppressed. These insights define a paradigm in G-protein signaling in which a pleiotropically acting modulator uses the same motif both to activate and to inhibit G proteins. Our findings also illuminate how such modulation of two opposing Gα proteins integrates downstream signals and cellular responses. PMID:27621449

Myosin IIA/IIB restrict adhesive and protrusive signaling to generate front-back polarity in migrating cells.

PubMed

Vicente-Manzanares, Miguel; Newell-Litwa, Karen; Bachir, Alexia I; Whitmore, Leanna A; Horwitz, Alan Rick

2011-04-18

Migratory front-back polarity emerges from the cooperative effect of myosin IIA (MIIA) and IIB (MIIB) on adhesive signaling. We demonstrate here that, during polarization, MIIA and MIIB coordinately promote localized actomyosin bundling, which generates large, stable adhesions that do not signal to Rac and thereby form the cell rear. MIIA formed dynamic actomyosin proto-bundles that mark the cell rear during spreading; it also bound to actin filament bundles associated with initial adhesion maturation in protrusions. Subsequent incorporation of MIIB stabilized the adhesions and actomyosin filaments with which it associated and formed a stable, extended rear. These adhesions did not turn over and no longer signal to Rac. Microtubules fine-tuned the polarity by positioning the front opposite the MIIA/MIIB-specified rear. Decreased Rac signaling in the vicinity of the MIIA/MIIB-stabilized proto-bundles and adhesions was accompanied by the loss of Rac guanine nucleotide exchange factor (GEFs), like βPIX and DOCK180, and by inhibited phosphorylation of key residues on adhesion proteins that recruit and activate Rac GEFs. These observations lead to a model for front-back polarity through local GEF depletion.

Environmental Land Management in Tajikistan

NASA Astrophysics Data System (ADS)

Makhmudov, Zafar; Ergashev, Murod

2015-04-01

Tackling Environmental Land Management in Tajikistan "Project approach" Khayrullo Ibodzoda, Zafar Mahmoudov, Murod Ergashev, Kamoliddin Abdulloev Among 28 countries in Europe and Central Asia, Tajikistan is estimated to be the most vulnerable to the climate change impacts depending on its high exposure and sensitivity combined with a very low adaptive capacity. The agricultural sector of Tajikistan is subject to lower and more erratic rainfalls, as well as dryness of water resources due to the possible temperature rising in the region, high evaporation, reducing the accumulation of snow in the mountain glaciers and increased frequency of extreme events. Climate change and variability are likely to pose certain risks, especially for those who prefer natural agriculture or pasture management that just reinforces the need for sound, adapted to new climatic conditions and improved principles of land management. Adoption of new strategies and best practices on sustainable land and water management for agricultural ecosystems will help the farmers and communities in addressing the abovementioned problems, adapt and become more resilient to changing climate by increasing wellbeing of local population, and contributing to food security and restoring productive natural resources. The Environmental Land Management and Rural Livelihoods Project is being financed by the Pilot Program for Climate Resilience (PPCR) and Global Environment Facility (GEF). The Project goal is to enable the rural population to increase their productive assets by improving management of natural resources and building resilience to climate change in selected climate vulnerable sites. The project will facilitate introduction of innovative measures on land use and agricultural production by providing small grants at the village level and grants for the Pasture User Groups (PUGs) at jamoat level in order to implement joint plans of pasture management and wellbred livestock, also for the Water User Associations (WUAs) to introduce sustainable on-farm water management practices. The Project comprises three components to be implemented in five years: 1. Rural Production and Land Resource Management Investments; 2. Knowledge Management and Institutional Support, and 3. Project Management and Coordination. These components include a set of grants from the PPCR and GEF that betrays the particular importance of the grant sources for the Project funding. This innovative combination of the PPCR and GEF grant funding will help in scheduling a scope of work under the Project and enable to implement certain activities on a pilot basis that otherwise could not be implemented at this level. Key partners are the Committee for Environmental Protection (Implementing Agency), the Ministry of Finance, the PPCR Secretariat in Tajikistan, Farkhor, Kulyab, Khovaling, Baljuvan, Tavildara and Jirgatal districts, the German Agency for International Development (GIZ) with its GREAT program which provides additional support to the community-based Project planning and institutional development, as well as technical agricultural advisory services. Currently the project has Project Implementation Group and most of its Facilitating Organizations in place that will carry out financial management, disbursements, procurement process, environmental management, social development, monitoring and evaluation. Workshops on coordinating the Project were held in the districts, as well as a series of Trainings of trainings and meetings were conducted for specialists and technical personnel. Next step is to initiate supporting local initiatives for climate adaptive land management and improved livelihoods based on Community Action Plans.

Ultrafast Self-Crystallization of High-External-Quantum-Efficient Fluoride Phosphors for Warm White Light-Emitting Diodes.

PubMed

Zhou, Wenli; Fang, Mu-Huai; Lian, Shixun; Liu, Ru-Shi

2018-05-30

In this study, we used HF (as good solvent) to dissolve K 2 GeF 6 and K 2 MnF 6 and added ethanol (as poor solvent) to cause ultrafast self-crystallization of K 2 GeF 6 :Mn 4+ crystals, which had an unprecedentedly high external quantum efficiency that reached 73%. By using the red phosphor, we achieved a high-quality warm white light-emitting diode with color-rendering index of R a = 94, R9 = 95, luminous efficacy of 150 lm W -1 , and correlated color temperature at 3652 K. Furthermore, the good-poor solvent strategy can be used to fast synthesize other fluorides.

Comparative Study of Multiplet Structures of Mn4+ in K2SiF6, K2GeF6, and K2TiF6 Based on First-Principles Configuration-Interaction Calculations

NASA Astrophysics Data System (ADS)

Novita, Mega; Ogasawara, Kazuyoshi

2012-02-01

We performed first-principles configuration-interaction calculations of multiplet energies for Mn4+ in K2SiF6, K2GeF6, and K2TiF6 crystals. The results indicate that corrections based on a single-electron calculation are effective for the prediction of 4A2 → 4T2 and 4A2 → 4T1a transition energies, while such corrections are not necessary for the prediction of the 4A2 → 2E transition energy. The cluster size dependence of the multiplet energies is small. However, the 4A2 → 2E transition energy is slightly improved by using larger clusters including K ions. The theoretical multiplet energies are improved further by considering the lattice relaxation effect. As a result, the characteristic multiplet energy shifts depending on the host crystal are well reproduced without using any empirical parameters. Although K2GeF6 and K2TiF6 have lower symmetry than K2SiF6, the results indicate that the variation of the multiplet energy is mainly determined by the Mn-F bond length.

Interplay between Solo and keratin filaments is crucial for mechanical force–induced stress fiber reinforcement

PubMed Central

Fujiwara, Sachiko; Ohashi, Kazumasa; Mashiko, Toshiya; Kondo, Hiroshi; Mizuno, Kensaku

2016-01-01

Mechanical force–induced cytoskeletal reorganization is essential for cell and tissue remodeling and homeostasis; however, the underlying cellular mechanisms remain elusive. Solo (ARHGEF40) is a RhoA-targeting guanine nucleotide exchange factor (GEF) involved in cyclical stretch–induced human endothelial cell reorientation and convergent extension cell movement in zebrafish gastrula. In this study, we show that Solo binds to keratin-8/keratin-18 (K8/K18) intermediate filaments through multiple sites. Solo overexpression promotes the formation of thick actin stress fibers and keratin bundles, whereas knockdown of Solo, expression of a GEF-inactive mutant of Solo, or inhibition of ROCK suppresses stress fiber formation and leads to disorganized keratin networks, indicating that the Solo-RhoA-ROCK pathway serves to precisely organize keratin networks, as well as to promote stress fibers. Of importance, knockdown of Solo or K18 or overexpression of GEF-inactive or deletion mutants of Solo suppresses tensile force–induced stress fiber reinforcement. Furthermore, knockdown of Solo or K18 suppresses tensile force-induced RhoA activation. These results strongly suggest that the interplay between Solo and K8/K18 filaments plays a crucial role in tensile force–induced RhoA activation and consequent actin cytoskeletal reinforcement. PMID:26823019

Conditional-suicide containment system for bacteria which mineralize aromatics

DOE Office of Scientific and Technical Information (OSTI.GOV)

Contreras, A.; Ramos, J.L.; Molin, S.

A model conditional-suicide system to control genetically engineered microorganisms able to degrade substituted benzoates is reported. The system is based on two elements. One element consists of a fusion between the promoter of the Pseudomonas putide TOL plasmid-encoded meta-cleavage pathway operon (P{sub m}) and the lacI gene encoding Lac repressor plus sylS, coding for the positive regulator of P{sub m}. The other element carries a fusion between the P{sub tac} promoter and the gef gene, which encodes a killing function. In the absence of effectors, expression of the P{sub tac}::gef cassette is no longer prevented and a high rate ofmore » cell killing is observed. The substitution of XylS for XylSthr45, a mutant regulator with altered effector specificity and increased affinity for benzoates, allows the control of populations able to degrade a wider range of benzoates at micromolar substrate concentrations. Given the wide effector specificity of the key regulators, the wild-type and mutant ZylS proteins, the system should allow the control of populations able to metabolize benzoate; methyl-, dimethyl-, chloro-, dichloro-, ethyl-, and methoxybenzoates; salicylate; and methyl- and chlorosalicylates. A small population of genetically engineered microorganisms became Gef resistant; however, the mechanism of such survival remains unknown.« less

National Centers for Environmental Prediction

Science.gov Websites

/ VISION | About EMC EMC > GEFS > IMPLEMENTATION SCHEDULLE Home Operational Products Experimental Data Verification Model Configuration Implementation Schedule Collaborators Documentation FAQ Code

PSD-Zip70 Deficiency Causes Prefrontal Hypofunction Associated with Glutamatergic Synapse Maturation Defects by Dysregulation of Rap2 Activity.

PubMed

Mayanagi, Taira; Yasuda, Hiroki; Sobue, Kenji

2015-10-21

Dysregulation of synapse formation and plasticity is closely related to the pathophysiology of psychiatric and neurodevelopmental disorders. The prefrontal cortex (PFC) is particularly important for executive functions such as working memory, cognition, and emotional control, which are impaired in the disorders. PSD-Zip70 (Lzts1/FEZ1) is a postsynaptic density (PSD) protein predominantly expressed in the frontal cortex, olfactory bulb, striatum, and hippocampus. Here we found that PSD-Zip70 knock-out (PSD-Zip70KO) mice exhibit working memory and cognitive defects, and enhanced anxiety-like behaviors. These abnormal behaviors are caused by impaired glutamatergic synapse transmission accompanied by tiny-headed immature dendritic spines in the PFC, due to aberrant Rap2 activation, which has roles in synapse formation and plasticity. PSD-Zip70 modulates the Rap2 activity by interacting with SPAR (spine-associated RapGAP) and PDZ-GEF1 (RapGEF) in the postsynapse. Furthermore, suppression of the aberrant Rap2 activation in the PFC rescued the behavioral defects in PSD-Zip70KO mice. Our data demonstrate a critical role for PSD-Zip70 in Rap2-dependent spine synapse development in the PFC and underscore the importance of this regulation in PFC-dependent behaviors. PSD-Zip70 deficiency causes behavioral defects in working memory and cognition, and enhanced anxiety due to prefrontal hypofunction. This study revealed that PSD-Zip70 plays essential roles in glutamatergic synapse maturation via modulation of the Rap2 activity in the PFC. PSD-Zip70 interacts with both SPAR (spine-associated RapGAP) and PDZ-GEF1 (RapGEF) and modulates the Rap2 activity in postsynaptic sites. Our results provide a novel Rap2-specific regulatory mechanism in synaptic maturation involving PSD-Zip70. Copyright © 2015 the authors 0270-6474/15/3514327-14$15.00/0.

SU-E-T-385: Evaluation of DVH Change for PTV Due to Patient Weight Loss in Prostate VMAT Using Gaussian Error Function

DOE Office of Scientific and Technical Information (OSTI.GOV)

Viraganathan, H; Jiang, R; Chow, J

Purpose: We proposed a method to predict the change of dose-volume histogram (DVH) for PTV due to patient weight loss in prostate volumetric modulated arc therapy (VMAT). This method is based on a pre-calculated patient dataset and DVH curve fitting using the Gaussian error function (GEF). Methods: Pre-calculated dose-volume data from patients having weight loss in prostate VMAT was employed to predict the change of PTV coverage due to reduced depth in external contour. The effect of patient weight loss in treatment was described by a prostate dose-volume factor (PDVF), which was evaluated by the prostate PTV. Along with themore » PDVF, the GEF was used to fit into the DVH curve for the PTV. To predict a new DVH due to weight loss, parameters from the GEF describing the shape of DVH curve were determined. Since the parameters were related to the PDVF as per the specific reduced depth, we could first predict the PDVF at a reduced depth based on the prostate size from the pre-calculated dataset. Then parameters of the GEF could be determined from the PDVF to plot the new DVH for the PTV corresponding to the reduced depth. Results: A MATLAB program was built basing on the patient dataset with different prostate sizes. We input data of the prostate size and reduced depth of the patient into the program. The program then calculated the PDVF and DVH for the PTV considering the patient weight loss. The program was verified by different patient cases with various reduced depths. Conclusion: Our method can estimate the change of DVH for the PTV due to patient weight loss quickly without CT rescan and replan. This would help the radiation staff to predict the change of PTV coverage, when patient’s external contour reduced in prostate VMAT.« less

Analytische Geometrie

NASA Astrophysics Data System (ADS)

Kemnitz, Arnfried

Der Grundgedanke der Analytischen Geometrie besteht darin, dass geometrische Untersuchungen mit rechnerischen Mitteln geführt werden. Geometrische Objekte werden dabei durch Gleichungen beschrieben und mit algebraischen Methoden untersucht.

Anomalous behaviour of thermodynamic properties at successive phase transitions in (NH4)3GeF7

NASA Astrophysics Data System (ADS)

Bogdanov, Evgeniy V.; Kartashev, Andrey V.; Pogoreltsev, Evgeniy I.; Gorev, Mikhail V.; Laptash, Natalia M.; Flerov, Igor N.

2017-12-01

Heat capacity, thermal dilatation, susceptibility to hydrostatic pressure and dielectric properties associated with succession of three phase transitions below room temperature in double fluoride salt (NH4)3GeF7 were studied. A possible transformation into the parent Pm-3m cubic phase was not observed up to the decomposition of compound. Nonferroelectric nature of structural distortions was confirmed. The DTA under pressure studies revealed a high temperature stability of two phases: P4/mbm and Pbam. The entropies of the phase transitions agree well with the model of structural distortions. Analysis of the thermal properties associated with the individual phase transitions in the framework of thermodynamic equations has shown a high reliability of the data obtained.

Evaluations of Extended-Range tropical Cyclone Forecasts in the Western North Pacific by using the Ensemble Reforecasts: Preliminary Results

NASA Astrophysics Data System (ADS)

Tsai, Hsiao-Chung; Chen, Pang-Cheng; Elsberry, Russell L.

2017-04-01

The objective of this study is to evaluate the predictability of the extended-range forecasts of tropical cyclone (TC) in the western North Pacific using reforecasts from National Centers for Environmental Prediction (NCEP) Global Ensemble Forecast System (GEFS) during 1996-2015, and from the Climate Forecast System (CFS) during 1999-2010. Tsai and Elsberry have demonstrated that an opportunity exists to support hydrological operations by using the extended-range TC formation and track forecasts in the western North Pacific from the ECMWF 32-day ensemble. To demonstrate this potential for the decision-making processes regarding water resource management and hydrological operation in Taiwan reservoir watershed areas, special attention is given to the skill of the NCEP GEFS and CFS models in predicting the TCs affecting the Taiwan area. The first objective of this study is to analyze the skill of NCEP GEFS and CFS TC forecasts and quantify the forecast uncertainties via verifications of categorical binary forecasts and probabilistic forecasts. The second objective is to investigate the relationships among the large-scale environmental factors [e.g., El Niño Southern Oscillation (ENSO), Madden-Julian Oscillation (MJO), etc.] and the model forecast errors by using the reforecasts. Preliminary results are indicating that the skill of the TC activity forecasts based on the raw forecasts can be further improved if the model biases are minimized by utilizing these reforecasts.

Identification of a Novel, Putative Rho-specific GDP/GTP Exchange Factor and a RhoA-binding Protein: Control of Neuronal Morphology

PubMed Central

Gebbink, Martijn F.B.G.; Kranenburg, Onno; Poland, Mieke; van Horck, Francis P.G.; Houssa, Brahim; Moolenaar, Wouter H.

1997-01-01

The small GTP-binding protein Rho has been implicated in the control of neuronal morphology. In N1E-115 neuronal cells, the Rho-inactivating C3 toxin stimulates neurite outgrowth and prevents actomyosin-based neurite retraction and cell rounding induced by lysophosphatidic acid (LPA), sphingosine-1-phosphate, or thrombin acting on their cognate G protein–coupled receptors. We have identified a novel putative GDP/GTP exchange factor, RhoGEF (190 kD), that interacts with both wild-type and activated RhoA, but not with Rac or Cdc42. RhoGEF, like activated RhoA, mimics receptor stimulation in inducing cell rounding and in preventing neurite outgrowth. Furthermore, we have identified a 116-kD protein, p116Rip, that interacts with both the GDP- and GTP-bound forms of RhoA in N1E-115 cells. Overexpression of p116Rip stimulates cell flattening and neurite outgrowth in a similar way to dominant-negative RhoA and C3 toxin. Cells overexpressing p116Rip fail to change their shape in response to LPA, as is observed after Rho inactivation. Our results indicate that (a) RhoGEF may link G protein–coupled receptors to RhoA activation and ensuing neurite retraction and cell rounding; and (b) p116Rip inhibits RhoA-stimulated contractility and promotes neurite outgrowth. PMID:9199174

Small-GTPase-associated signaling by the guanine nucleotide exchange factors CpDock180 and CpCdc24, the GTPase effector CpSte20, and the scaffold protein CpBem1 in Claviceps purpurea.

PubMed

Herrmann, Andrea; Tillmann, Britta A M; Schürmann, Janine; Bölker, Michael; Tudzynski, Paul

2014-04-01

Monomeric GTPases of the Rho subfamily are important mediators of polar growth and NADPH (Nox) signaling in a variety of organisms. These pathways influence the ability of Claviceps purpurea to infect host plants. GTPase regulators contribute to the nucleotide loading cycle that is essential for proper functionality of the GTPases. Scaffold proteins gather GTPase complexes to facilitate proper function. The guanine nucleotide exchange factors (GEFs) CpCdc24 and CpDock180 activate GTPase signaling by triggering nucleotide exchange of the GTPases. Here we show that CpCdc24 harbors nucleotide exchange activity for both Rac and Cdc42 homologues. The GEFs partly share the cellular distribution of the GTPases and interact with the putative upstream GTPase CpRas1. Interaction studies show the formation of higher-order protein complexes, mediated by the scaffold protein CpBem1. Besides the GTPases and GEFs, these complexes also contain the GTPase effectors CpSte20 and CpCla4, as well as the regulatory protein CpNoxR. Functional characterizations suggest a role of CpCdc24 mainly in polarity, whereas CpDock180 is involved in stress tolerance mechanisms. These findings indicate the dynamic formation of small GTPase complexes and improve the model for GTPase-associated signaling in C. purpurea.

A dual role for the RhoGEF Ephexin5 in regulation of dendritic spine outgrowth

PubMed Central

Hamilton, AM; Lambert, JT; Parajuli, LK; Vivas, O; Park, DK; Stein, IS; Jahncke, JN; Greenberg, ME; Margolis, SS; Zito, K

2017-01-01

The outgrowth of new dendritic spines is closely linked to the formation of new synapses, and is thought to be a vital component of the experience-dependent circuit plasticity that supports learning. Here, we examined the role of the RhoGEF Ephexin5 in driving activity-dependent spine outgrowth. We found that reducing Ephexin5 levels increased spine outgrowth, and increasing Ephexin5 levels decreased spine outgrowth in a GEF-dependent manner, suggesting that Ephexin5 acts as an inhibitor of spine outgrowth. Notably, we found that increased neural activity led to a proteasome-dependent reduction in the levels of Ephexin5 in neuronal dendrites, which could facilitate the enhanced spine outgrowth observed following increased neural activity. Surprisingly, we also found that Ephexin5-GFP levels were elevated on the dendrite at sites of future new spines, prior to new spine outgrowth. Moreover, lowering neuronal Ephexin5 levels inhibited new spine outgrowth in response to both global increases in neural activity and local glutamatergic stimulation of the dendrite, suggesting that Ephexin5 is necessary for activity-dependent spine outgrowth. Our data support a model in which Ephexin5 serves a dual role in spinogenesis, acting both as a brake on overall spine outgrowth and as a necessary component in the site-specific formation of new spines. PMID:28185854

A dual role for the RhoGEF Ephexin5 in regulation of dendritic spine outgrowth.

PubMed

Hamilton, A M; Lambert, J T; Parajuli, L K; Vivas, O; Park, D K; Stein, I S; Jahncke, J N; Greenberg, M E; Margolis, S S; Zito, K

2017-04-01

The outgrowth of new dendritic spines is closely linked to the formation of new synapses, and is thought to be a vital component of the experience-dependent circuit plasticity that supports learning. Here, we examined the role of the RhoGEF Ephexin5 in driving activity-dependent spine outgrowth. We found that reducing Ephexin5 levels increased spine outgrowth, and increasing Ephexin5 levels decreased spine outgrowth in a GEF-dependent manner, suggesting that Ephexin5 acts as an inhibitor of spine outgrowth. Notably, we found that increased neural activity led to a proteasome-dependent reduction in the levels of Ephexin5 in neuronal dendrites, which could facilitate the enhanced spine outgrowth observed following increased neural activity. Surprisingly, we also found that Ephexin5-GFP levels were elevated on the dendrite at sites of future new spines, prior to new spine outgrowth. Moreover, lowering neuronal Ephexin5 levels inhibited new spine outgrowth in response to both global increases in neural activity and local glutamatergic stimulation of the dendrite, suggesting that Ephexin5 is necessary for activity-dependent spine outgrowth. Our data support a model in which Ephexin5 serves a dual role in spinogenesis, acting both as a brake on overall spine outgrowth and as a necessary component in the site-specific formation of new spines. Copyright © 2017 Elsevier Inc. All rights reserved.

Phosphorylation-dependent Regulation of Connecdenn/DENND1 Guanine Nucleotide Exchange Factors*

PubMed Central

Kulasekaran, Gopinath; Nossova, Nadya; Marat, Andrea L.; Lund, Ingrid; Cremer, Christopher; Ioannou, Maria S.; McPherson, Peter S.

2015-01-01

Connecdenn 1/2 are DENN (differentially expressed in normal and neoplastic cells) domain-bearing proteins that function as GEFs (guanine nucleotide exchange factors) for the small GTPase Rab35. Disruption of connecdenn/Rab35 function leads to defects in the recycling of multiple cargo proteins from endosomes with altered cell function, yet the regulation of connecdenn GEF activity is unexplored. We now demonstrate that connecdenn 1/2 are autoinhibited such that the purified, full-length proteins have significantly less Rab35 binding and GEF activity than the isolated DENN domain. Both proteins are phosphorylated with prominent phosphorylation sites between residues 500 and 600 of connecdenn 1. A large scale proteomics screen revealed that connecdenn 1 is phosphorylated at residues Ser-536 and Ser-538 in an Akt-dependent manner in response to insulin stimulation of adipocytes. Interestingly, we find that an Akt inhibitor reduces connecdenn 1 interaction with Rab35 after insulin treatment of adipocytes. Remarkably, a peptide flanking Ser-536/Ser-538 binds the DENN domain of connecdenn 1, whereas a phosphomimetic peptide does not. Moreover, connecdenn 1 interacts with 14-3-3 proteins, and this interaction is also disrupted by Akt inhibition and by mutation of Ser-536/Ser-538. We propose that Akt phosphorylation of connecdenn 1 downstream of insulin activation regulates connecdenn 1 function through an intramolecular interaction. PMID:26055712

Gamma and x-ray irradiation effects on different Ge and Ge/F doped optical fibers

DOE Office of Scientific and Technical Information (OSTI.GOV)

Alessi, A., E-mail: antonino.alessi@univ-st-etienne.fr; Girard, S.; Di Francesca, D.

2015-08-28

We performed electron paramagnetic resonance (EPR) measurements on γ and X ray irradiated Ge doped and Ge/F co-doped optical fibers. We considered three different drawing conditions (speed and tension), and for each type of drawing, we studied Ge and Ge/F doped samples having Ge doping level above 4% by weight. The EPR data recorded for the γ ray irradiated fibers confirm that all the samples exhibit a very close radiation response regardless of the drawing conditions corresponding to values used for the production of specialty fibers. Furthermore, as for the X irradiated materials, in the γ ray irradiated F co-dopedmore » fibers, we observed that the Ge(1) and the Ge(2) defects generation is unchanged, whereas it was enhanced for the E'Ge. In the various fibers, the comparison of the γ and X-ray induced concentrations of these kinds of Ge related defects indicates that the two irradiations induce similar effects regardless of the different employed dose rates and sources. Confocal microscopy luminescence results show that the starting content of the Germanium Lone Pair Center (GLPC) is neither strongly affected by the Ge content nor by the drawing conditions, and we consider the similarity of the GLPC content as key factor in determining many of the above reported similarities.« less

Mechanism of the Exchange Reaction in HRAS from Multiscale Modeling

PubMed Central

Kapoor, Abhijeet; Travesset, Alex

2014-01-01

HRAS regulates cell growth promoting signaling processes by cycling between active (GTP-bound) and inactive (GDP-bound) states. Understanding the transition mechanism is central for the design of small molecules to inhibit the formation of RAS-driven tumors. Using a multiscale approach involving coarse-grained (CG) simulations, all-atom classical molecular dynamics (CMD; total of 3.02 µs), and steered molecular dynamics (SMD) in combination with Principal Component Analysis (PCA), we identified the structural features that determine the nucleotide (GDP) exchange reaction. We show that weakening the coupling between the SwitchI (residues 25–40) and SwitchII (residues 59–75) accelerates the opening of SwitchI; however, an open conformation of SwitchI is unstable in the absence of guanine nucleotide exchange factors (GEFs) and rises up towards the bound nucleotide to close the nucleotide pocket. Both I21 and Y32, play a crucial role in SwitchI transition. We show that an open SwitchI conformation is not necessary for GDP destabilization but is required for GDP/Mg escape from the HRAS. Further, we present the first simulation study showing displacement of GDP/Mg away from the nucleotide pocket. Both SwitchI and SwitchII, delays the escape of displaced GDP/Mg in the absence of GEF. Based on these results, a model for the mechanism of GEF in accelerating the exchange process is hypothesized. PMID:25272152

National Centers for Environmental Prediction

Science.gov Websites

---------------------------------------------------------------------------------------------------------------------------------------------------- IITM CFS v2 Forecast for 2017 monsoon : link Experimental short-range GEFS ensemble forecast : link Experimental short-range GFS-T1534(upto 8days) forecast : link

Machineries regulating the activity of the small GTPase Arf6 in cancer cells are potential targets for developing innovative anti-cancer drugs.

PubMed

Yamauchi, Yohei; Miura, Yuki; Kanaho, Yasunori

2017-01-01

The Small GTPase ADP-ribosylation factor 6 (Arf6) functions as the molecular switch in cellular signaling pathways by cycling between GDP-bound inactive and GTP-bound active form, which is precisely regulated by two regulators, guanine nucleotide exchange factors (GEFs) and GTPase-activating proteins (GAPs). Numerous studies have shown that these machineries play critical roles in tumor angiogenesis/growth and cancer cell invasion/metastasis through regulating the cycling of Arf6. Here, we summarize accumulating knowledge for involvement of Arf6 GEFs/GAPs and small molecule inhibitors of Arf6 signaling/cycling in cancer progression, and discuss possible strategies for developing innovative anti-cancer drugs targeting Arf6 signaling/cycling. Copyright © 2016 Elsevier Ltd. All rights reserved.

General Description of Fission Observables: GEF Model Code

DOE Office of Scientific and Technical Information (OSTI.GOV)

Schmidt, K.-H.; Jurado, B., E-mail: jurado@cenbg.in2p3.fr; Amouroux, C.

2016-01-15

The GEF (“GEneral description of Fission observables”) model code is documented. It describes the observables for spontaneous fission, neutron-induced fission and, more generally, for fission of a compound nucleus from any other entrance channel, with given excitation energy and angular momentum. The GEF model is applicable for a wide range of isotopes from Z = 80 to Z = 112 and beyond, up to excitation energies of about 100 MeV. The results of the GEF model are compared with fission barriers, fission probabilities, fission-fragment mass- and nuclide distributions, isomeric ratios, total kinetic energies, and prompt-neutron and prompt-gamma yields and energymore » spectra from neutron-induced and spontaneous fission. Derived properties of delayed neutrons and decay heat are also considered. The GEF model is based on a general approach to nuclear fission that explains a great part of the complex appearance of fission observables on the basis of fundamental laws of physics and general properties of microscopic systems and mathematical objects. The topographic theorem is used to estimate the fission-barrier heights from theoretical macroscopic saddle-point and ground-state masses and experimental ground-state masses. Motivated by the theoretically predicted early localisation of nucleonic wave functions in a necked-in shape, the properties of the relevant fragment shells are extracted. These are used to determine the depths and the widths of the fission valleys corresponding to the different fission channels and to describe the fission-fragment distributions and deformations at scission by a statistical approach. A modified composite nuclear-level-density formula is proposed. It respects some features in the superfluid regime that are in accordance with new experimental findings and with theoretical expectations. These are a constant-temperature behaviour that is consistent with a considerably increased heat capacity and an increased pairing condensation energy that is consistent with the collective enhancement of the level density. The exchange of excitation energy and nucleons between the nascent fragments on the way from saddle to scission is estimated according to statistical mechanics. As a result, excitation energy and unpaired nucleons are predominantly transferred to the heavy fragment in the superfluid regime. This description reproduces some rather peculiar observed features of the prompt-neutron multiplicities and of the even-odd effect in fission-fragment Z distributions. For completeness, some conventional descriptions are used for calculating pre-equilibrium emission, fission probabilities and statistical emission of neutrons and gamma radiation from the excited fragments. Preference is given to simple models that can also be applied to exotic nuclei compared to more sophisticated models that need precise empirical input of nuclear properties, e.g. spectroscopic information. The approach reveals a high degree of regularity and provides a considerable insight into the physics of the fission process. Fission observables can be calculated with a precision that complies with the needs for applications in nuclear technology without specific adjustments to measured data of individual systems. The GEF executable runs out of the box with no need for entering any empirical data. This unique feature is of valuable importance, because the number of systems and energies of potential significance for fundamental and applied science will never be possible to be measured. The relevance of the approach for examining the consistency of experimental results and for evaluating nuclear data is demonstrated.« less

National Centers for Environmental Prediction

Science.gov Websites

SYSTEM CFS CLIMATE FORECAST SYSTEM NAQFC NAQFC MODEL GEFS GLOBAL ENSEMBLE FORECAST SYSTEM HWRF HURRICANE WEATHER RESEARCH and FORECASTING HMON HMON - OPERATIONAL HURRICANE FORECASTING WAVEWATCH III WAVEWATCH III

PDZ Protein Regulation of G Protein-Coupled Receptor Trafficking and Signaling Pathways.

PubMed

Dunn, Henry A; Ferguson, Stephen S G

2015-10-01

G protein-coupled receptors (GPCRs) contribute to the regulation of every aspect of human physiology and are therapeutic targets for the treatment of numerous diseases. As a consequence, understanding the myriad of mechanisms controlling GPCR signaling and trafficking is essential for the development of new pharmacological strategies for the treatment of human pathologies. Of the many GPCR-interacting proteins, postsynaptic density protein of 95 kilodaltons, disc large, zona occludens-1 (PDZ) domain-containing proteins appear most abundant and have similarly been implicated in disease mechanisms. PDZ proteins play an important role in regulating receptor and channel protein localization within synapses and tight junctions and function to scaffold intracellular signaling protein complexes. In the current study, we review the known functional interactions between PDZ domain-containing proteins and GPCRs and provide insight into the potential mechanisms of action. These PDZ domain-containing proteins include the membrane-associated guanylate-like kinases [postsynaptic density protein of 95 kilodaltons; synapse-associated protein of 97 kilodaltons; postsynaptic density protein of 93 kilodaltons; synapse-associated protein of 102 kilodaltons; discs, large homolog 5; caspase activation and recruitment domain and membrane-associated guanylate-like kinase domain-containing protein 3; membrane protein, palmitoylated 3; calcium/calmodulin-dependent serine protein kinase; membrane-associated guanylate kinase protein (MAGI)-1, MAGI-2, and MAGI-3], Na(+)/H(+) exchanger regulatory factor proteins (NHERFs) (NHERF1, NHERF2, PDZ domain-containing kidney protein 1, and PDZ domain-containing kidney protein 2), Golgi-associated PDZ proteins (Gα-binding protein interacting protein, C-terminus and CFTR-associated ligand), PDZ domain-containing guanine nucleotide exchange factors (GEFs) 1 and 2, regulator of G protein signaling (RGS)-homology-RhoGEFs (PDZ domain-containing RhoGEF and leukemia-associated RhoGEF), RGS3 and RGS12, spinophilin and neurabin-1, SRC homology 3 domain and multiple ankyrin repeat domain (Shank) proteins (Shank1, Shank2, and Shank3), partitioning defective proteins 3 and 6, multiple PDZ protein 1, Tamalin, neuronal nitric oxide synthase, syntrophins, protein interacting with protein kinase C α 1, syntenin-1, and sorting nexin 27. Copyright © 2015 by The American Society for Pharmacology and Experimental Therapeutics.

Regulation of neurite morphogenesis by interaction between R7 regulator of G protein signaling complexes and G protein subunit Gα13.

PubMed

Scherer, Stephanie L; Cain, Matthew D; Kanai, Stanley M; Kaltenbronn, Kevin M; Blumer, Kendall J

2017-06-16

The R7 regulator of G protein signaling family (R7-RGS) critically regulates nervous system development and function. Mice lacking all R7-RGS subtypes exhibit diverse neurological phenotypes, and humans bearing mutations in the retinal R7-RGS isoform RGS9-1 have vision deficits. Although each R7-RGS subtype forms heterotrimeric complexes with Gβ 5 and R7-RGS-binding protein (R7BP) that regulate G protein-coupled receptor signaling by accelerating deactivation of G i/o α-subunits, several neurological phenotypes of R7-RGS knock-out mice are not readily explained by dysregulated G i/o signaling. Accordingly, we used tandem affinity purification and LC-MS/MS to search for novel proteins that interact with R7-RGS heterotrimers in the mouse brain. Among several proteins detected, we focused on Gα 13 because it had not been linked to R7-RGS complexes before. Split-luciferase complementation assays indicated that Gα 13 in its active or inactive state interacts with R7-RGS heterotrimers containing any R7-RGS isoform. LARG (leukemia-associated Rho guanine nucleotide exchange factor (GEF)), PDZ-RhoGEF, and p115RhoGEF augmented interaction between activated Gα 13 and R7-RGS heterotrimers, indicating that these effector RhoGEFs can engage Gα 13 ·R7-RGS complexes. Because Gα 13 /R7-RGS interaction required R7BP, we analyzed phenotypes of neuronal cell lines expressing RGS7 and Gβ 5 with or without R7BP. We found that neurite retraction evoked by Gα 12/13 -dependent lysophosphatidic acid receptors was augmented in R7BP-expressing cells. R7BP expression blunted neurite formation evoked by serum starvation by signaling mechanisms involving Gα 12/13 but not Gα i/o These findings provide the first evidence that R7-RGS heterotrimers interact with Gα 13 to augment signaling pathways that regulate neurite morphogenesis. This mechanism expands the diversity of functions whereby R7-RGS complexes regulate critical aspects of nervous system development and function. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

An Assessment of the Subseasonal Forecast Performance in the Extended Global Ensemble Forecast System (GEFS)

NASA Astrophysics Data System (ADS)

Sinsky, E.; Zhu, Y.; Li, W.; Guan, H.; Melhauser, C.

2017-12-01

Optimal forecast quality is crucial for the preservation of life and property. Improving monthly forecast performance over both the tropics and extra-tropics requires attention to various physical aspects such as the representation of the underlying SST, model physics and the representation of the model physics uncertainty for an ensemble forecast system. This work focuses on the impact of stochastic physics, SST and the convection scheme on forecast performance for the sub-seasonal scale over the tropics and extra-tropics with emphasis on the Madden-Julian Oscillation (MJO). A 2-year period is evaluated using the National Centers for Environmental Prediction (NCEP) Global Ensemble Forecast System (GEFS). Three experiments with different configurations than the operational GEFS were performed to illustrate the impact of the stochastic physics, SST and convection scheme. These experiments are compared against a control experiment (CTL) which consists of the operational GEFS but its integration is extended from 16 to 35 days. The three configurations are: 1) SPs, which uses a Stochastically Perturbed Physics Tendencies (SPPT), Stochastic Perturbed Humidity (SHUM) and Stochastic Kinetic Energy Backscatter (SKEB); 2) SPs+SST_bc, which uses a combination of SPs and a bias-corrected forecast SST from the NCEP Climate Forecast System Version 2 (CFSv2); and 3) SPs+SST_bc+SA_CV, which combines SPs, a bias-corrected forecast SST and a scale aware convection scheme. When comparing to the CTL experiment, SPs shows substantial improvement. The MJO skill has improved by about 4 lead days during the 2-year period. Improvement is also seen over the extra-tropics due to the updated stochastic physics, where there is a 3.1% and a 4.2% improvement during weeks 3 and 4 over the northern hemisphere and southern hemisphere, respectively. Improvement is also seen when the bias-corrected CFSv2 SST is combined with SPs. Additionally, forecast performance enhances when the scale aware convection scheme (SPs+SST_bc+SA_CV) is added, especially over the tropics. Among the three experiments, the SPs+SST_bc+SA_CV is the best configuration in MJO forecast skill.

Ion chemistry in germane/fluorocompounds gaseous mixtures: a mass spectrometric and theoretical study.

PubMed

Antoniotti, Paola; Rabezzana, Roberto; Turco, Francesca; Borocci, Stefano; Giordani, Maria; Grandinetti, Felice

2008-10-01

The ion-molecule reactions occurring in GeH(4)/NF(3), GeH(4)/SF(6), and GeH(4)/SiF(4) gaseous mixtures have been investigated by ion trap mass spectrometry and ab initio calculations. While the NF(x)(+) (x=1-3) react with GeH(4) mainly by the exothermic charge transfer, the open-shell Ge(+) and GeH(2)(+) undergo the efficient F-atom abstraction from NF(3) and form GeF(+) and F-GeH(2)(+) as the only ionic products. The mechanisms of these two processes are quite similar and involve the formation of the fluorine-coordinated complexes Ge-F-NF(2)(+) and H(2)Ge-F-NF(2)(+), their subsequent crossing to the significantly more stable isomers FGe-NF(2)(+) and F-GeH(2)-NF(2)(+), and the eventual dissociation of these ions into GeF(+) (or F-GeH(2)(+)) and NF(2). The closed-shell GeH(+) and GeH(3)(+) are instead much less reactive towards NF(3), and the only observed process is the less efficient formation of GeF(+) from GeH(+). The theoretical investigation of this unusual H/F exchange reaction suggests the involvement of vibrationally-hot GeH(+). Passing from NF(3) to SF(6) and SiF(4), the average strength of the M-F bond increases from 70 to 79 and 142 kcal mol(-1), and in fact the only process observed by reacting GeH(n)(+) (n=0-3) with SF(6) and SiF(4) is the little efficient F-atom abstraction from SF(6) by Ge(+). Irrespective of the experimental conditions, we did not observe any ionic product of Ge-N, Ge-S, or Ge-Si connectivity. This is in line with the previously observed exclusive formation of GeF(+) from the reaction between Ge(+) and C-F compounds such as CH(3)F. Additionally observed processes include in particular the conceivable formation of the elusive thiohypofluorous acid FSH from the reaction between SF(+) and GeH(4).

Mapping the functional versatility and fragility of Ras GTPase signaling circuits through in vitro network reconstitution.

PubMed

Coyle, Scott M; Lim, Wendell A

2016-01-14

The Ras-superfamily GTPases are central controllers of cell proliferation and morphology. Ras signaling is mediated by a system of interacting molecules: upstream enzymes (GEF/GAP) regulate Ras's ability to recruit multiple competing downstream effectors. We developed a multiplexed, multi-turnover assay for measuring the dynamic signaling behavior of in vitro reconstituted H-Ras signaling systems. By including both upstream regulators and downstream effectors, we can systematically map how different network configurations shape the dynamic system response. The concentration and identity of both upstream and downstream signaling components strongly impacted the timing, duration, shape, and amplitude of effector outputs. The distorted output of oncogenic alleles of Ras was highly dependent on the balance of positive (GAP) and negative (GEF) regulators in the system. We found that different effectors interpreted the same inputs with distinct output dynamics, enabling a Ras system to encode multiple unique temporal outputs in response to a single input. We also found that different Ras-to-GEF positive feedback mechanisms could reshape output dynamics in distinct ways, such as signal amplification or overshoot minimization. Mapping of the space of output behaviors accessible to Ras provides a design manual for programming Ras circuits, and reveals how these systems are readily adapted to produce an array of dynamic signaling behaviors. Nonetheless, this versatility comes with a trade-off of fragility, as there exist numerous paths to altered signaling behaviors that could cause disease.

Solo/Trio8, a membrane-associated short isoform of Trio, modulates endosome dynamics and neurite elongation.

PubMed

Sun, Ying-Jie; Nishikawa, Kaori; Yuda, Hideki; Wang, Yu-Lai; Osaka, Hitoshi; Fukazawa, Nobuna; Naito, Akira; Kudo, Yoshihisa; Wada, Keiji; Aoki, Shunsuke

2006-09-01

With DNA microarrays, we identified a gene, termed Solo, that is downregulated in the cerebellum of Purkinje cell degeneration mutant mice. Solo is a mouse homologue of rat Trio8-one of multiple Trio isoforms recently identified in rat brain. Solo/Trio8 contains N-terminal sec14-like and spectrin-like repeat domains followed by a single guanine nucleotide exchange factor 1 (GEF1) domain, but it lacks the C-terminal GEF2, immunoglobulin-like, and kinase domains that are typical of Trio. Solo/Trio8 is predominantly expressed in Purkinje neurons of the mouse brain, and expression begins following birth and increases during Purkinje neuron maturation. We identified a novel C-terminal membrane-anchoring domain in Solo/Trio8 that is required for enhanced green fluorescent protein-Solo/Trio8 localization to early endosomes (positive for both early-endosome antigen 1 [EEA1] and Rab5) in COS-7 cells and primary cultured neurons. Solo/Trio8 overexpression in COS-7 cells augmented the EEA1-positive early-endosome pool, and this effect was abolished via mutation and inactivation of the GEF domain or deletion of the C-terminal membrane-anchoring domain. Moreover, primary cultured neurons transfected with Solo/Trio8 showed increased neurite elongation that was dependent on these domains. These results suggest that Solo/Trio8 acts as an early-endosome-specific upstream activator of Rho family GTPases for neurite elongation of developing Purkinje neurons.

Evaluating the influence of geo-environmental factors on gully erosion in a semi-arid region of Iran: An integrated framework.

PubMed

Rahmati, Omid; Tahmasebipour, Naser; Haghizadeh, Ali; Pourghasemi, Hamid Reza; Feizizadeh, Bakhtiar

2017-02-01

Despite the importance of soil erosion in sustainable development goals in arid and semi-arid areas, the study of the geo-environmental conditions and factors influencing gully erosion occurrence is rarely undertaken. As effort to this challenge, the main objective of this study is to apply an integrated approach of Geographic Object-Based Image Analysis (GEOBIA) together with high-spatial resolution imagery (SPOT-5) for detecting gully erosion features at the Kashkan-Poldokhtar watershed, Iran. We also aimed to apply a Conditional Probability (CP) model for establishing the spatial relationship between gullies and the Geo-Environmental Factors (GEFs). The gully erosion inventory map prepared using GEOBIA and field surveying was randomly partitioned into two subsets: (1) part 1 that contains 70% was used in the training phase of the CP model; (2) part 2 is a validation dataset (30%) for validation of the model and to confirm its accuracy. Prediction performances of the GEOBIA and CP model were checked by overall accuracy and Receiver Operating Characteristics (ROC) curve methods, respectively. In addition, the influence of all GEFs on gully erosion was evaluated by performing a sensitivity analysis model. The validation findings illustrated that overall accuracy for GEOBIA approach and the area under the ROC curve for the CP model were 92.4% and 89.9%, respectively. Also, based on sensitivity analysis, soil texture, drainage density, and lithology represent significantly effects on the gully erosion occurrence. This study has shown that the integrated framework can be successfully used for modeling gully erosion occurrence in a data-poor environment. Copyright © 2016 Elsevier B.V. All rights reserved.

Production of Sn and Sb isotopes in high-energy neutron-induced fission of natU

NASA Astrophysics Data System (ADS)

Mattera, A.; Pomp, S.; Lantz, M.; Rakopoulos, V.; Solders, A.; Al-Adili, A.; Penttilä, H.; Moore, I. D.; Rinta-Antila, S.; Eronen, T.; Kankainen, A.; Pohjalainen, I.; Gorelov, D.; Canete, L.; Nesterenko, D.; Vilén, M.; Äystö, J.

2018-03-01

The first systematic measurement of neutron-induced fission yields has been performed at the upgraded IGISOL-4 facility at the University of Jyväskylä, Finland. The fission products from high-energy neutron-induced fission of nat U were stopped in a gas cell filled with helium buffer gas, and were online separated with a dipole magnet. The isobars, with masses in the range A = 128-133 , were transported to a tape-implantation station and identified using γ -spectroscopy. We report here the relative cumulative isotopic yields of tin ( Z = 50) and the relative independent isotopic yields of antimony ( Z = 51) . Isomeric yield ratios were also obtained for five nuclides. The yields of tin show a staggered behaviour around A = 131 , not observed in the ENDF/B-VII.1 evaluation. The yields of antimony also contradict the trend from the evaluation, but are in agreement with a calculation performed using the GEF model that shows the yield increasing with mass in the range A = 128-133.

RhoGTPase signalling at epithelial tight junctions: Bridging the GAP between polarity and cancer.

PubMed

Zihni, Ceniz; Terry, Stephen James

2015-07-01

The establishment and maintenance of epithelial polarity must be correctly controlled for normal development and homeostasis. Tight junctions (TJ) in vertebrates define apical and basolateral membrane domains in polarized epithelia via bi-directional, complex signalling pathways between TJ themselves and the cytoskeleton they are associated with. RhoGTPases are central to these processes and evidence suggests that their regulation is coordinated by interactions between GEFs and GAPs with junctional, cytoplasmic adapter proteins. In this InFocus review we determine that the expression, localization or stability of a variety of these adaptor proteins is altered in various cancers, potentially representing an important mechanistic link between loss of polarity and cancer. We focus here, on two well characterized RhoGTPases Cdc42 and RhoA who's GEFs and GAPs are predominantly localized to TJ via cytoplasmic adaptor proteins. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

Mapping the functional versatility and fragility of Ras GTPase signaling circuits through in vitro network reconstitution

PubMed Central

Coyle, Scott M; Lim, Wendell A

2016-01-01

The Ras-superfamily GTPases are central controllers of cell proliferation and morphology. Ras signaling is mediated by a system of interacting molecules: upstream enzymes (GEF/GAP) regulate Ras’s ability to recruit multiple competing downstream effectors. We developed a multiplexed, multi-turnover assay for measuring the dynamic signaling behavior of in vitro reconstituted H-Ras signaling systems. By including both upstream regulators and downstream effectors, we can systematically map how different network configurations shape the dynamic system response. The concentration and identity of both upstream and downstream signaling components strongly impacted the timing, duration, shape, and amplitude of effector outputs. The distorted output of oncogenic alleles of Ras was highly dependent on the balance of positive (GAP) and negative (GEF) regulators in the system. We found that different effectors interpreted the same inputs with distinct output dynamics, enabling a Ras system to encode multiple unique temporal outputs in response to a single input. We also found that different Ras-to-GEF positive feedback mechanisms could reshape output dynamics in distinct ways, such as signal amplification or overshoot minimization. Mapping of the space of output behaviors accessible to Ras provides a design manual for programming Ras circuits, and reveals how these systems are readily adapted to produce an array of dynamic signaling behaviors. Nonetheless, this versatility comes with a trade-off of fragility, as there exist numerous paths to altered signaling behaviors that could cause disease. DOI: http://dx.doi.org/10.7554/eLife.12435.001 PMID:26765565

The R-Ras/RIN2/Rab5 complex controls endothelial cell adhesion and morphogenesis via active integrin endocytosis and Rac signaling

PubMed Central

Sandri, Chiara; Caccavari, Francesca; Valdembri, Donatella; Camillo, Chiara; Veltel, Stefan; Santambrogio, Martina; Lanzetti, Letizia; Bussolino, Federico; Ivaska, Johanna; Serini, Guido

2012-01-01

During developmental and tumor angiogenesis, semaphorins regulate blood vessel navigation by signaling through plexin receptors that inhibit the R-Ras subfamily of small GTPases. R-Ras is mainly expressed in vascular cells, where it induces adhesion to the extracellular matrix (ECM) through unknown mechanisms. We identify the Ras and Rab5 interacting protein RIN2 as a key effector that in endothelial cells interacts with and mediates the pro-adhesive and -angiogenic activity of R-Ras. Both R-Ras-GTP and RIN2 localize at nascent ECM adhesion sites associated with lamellipodia. Upon binding, GTP-loaded R-Ras converts RIN2 from a Rab5 guanine nucleotide exchange factor (GEF) to an adaptor that first interacts at high affinity with Rab5-GTP to promote the selective endocytosis of ligand-bound/active β1 integrins and then causes the translocation of R-Ras to early endosomes. Here, the R-Ras/RIN2/Rab5 signaling module activates Rac1-dependent cell adhesion via TIAM1, a Rac GEF that localizes on early endosomes and is stimulated by the interaction with both Ras proteins and the vesicular lipid phosphatidylinositol 3-monophosphate. In conclusion, the ability of R-Ras-GTP to convert RIN2 from a GEF to an adaptor that preferentially binds Rab5-GTP allows the triggering of the endocytosis of ECM-bound/active β1 integrins and the ensuing funneling of R-Ras-GTP toward early endosomes to elicit the pro-adhesive and TIAM1-mediated activation of Rac1. PMID:22825554

FgBud3, a Rho4-Interacting Guanine Nucleotide Exchange Factor, Is Involved in Polarity Growth, Cell Division and Pathogenicity of Fusarium graminearum.

PubMed

Zhang, Chengkang; Luo, Zenghong; He, Dongdong; Su, Li; Yin, Hui; Wang, Guo; Liu, Hong; Rensing, Christopher; Wang, Zonghua

2018-01-01

Rho GTPases are signaling macromolecules that are associated with developmental progression and pathogenesis of Fusarium graminearum . Generally, enzymatic activities of Rho GTPases are regulated by Rho GTPase guanine nucleotide exchange factors (RhoGEFs). In this study, we identified a putative RhoGEF encoding gene ( FgBUD3 ) in F. graminearum database and proceeded further by using a functional genetic approach to generate FgBUD3 targeted gene deletion mutant. Phenotypic analysis results showed that the deletion of FgBUD3 caused severe reduction in growth of FgBUD3 mutant generated during this study. We also observed that the deletion of FgBUD3 completely abolished sexual reproduction and triggered the production of abnormal asexual spores with nearly no septum in ΔFgbud3 strain. Further results obtained from infection assays conducted during this research revealed that the FgBUD3 defective mutant lost its pathogenicity on wheat and hence, suggests FgBud3 plays an essential role in the pathogenicity of F. graminearum . Additional, results derived from yeast two-hybrid assays revealed that FgBud3 strongly interacted with FgRho4 compared to the interaction with FgRho2, FgRho3, and FgCdc42. Moreover, we found that FgBud3 interacted with both GTP-bound and GDP-bound form of FgRho4. From these results, we subsequently concluded that, the Rho4-interacting GEF protein FgBud3 crucially promotes vegetative growth, asexual and sexual development, cell division and pathogenicity in F. graminearum .

Interplay between Solo and keratin filaments is crucial for mechanical force-induced stress fiber reinforcement.

PubMed

Fujiwara, Sachiko; Ohashi, Kazumasa; Mashiko, Toshiya; Kondo, Hiroshi; Mizuno, Kensaku

2016-03-15

Mechanical force-induced cytoskeletal reorganization is essential for cell and tissue remodeling and homeostasis; however, the underlying cellular mechanisms remain elusive. Solo (ARHGEF40) is a RhoA-targeting guanine nucleotide exchange factor (GEF) involved in cyclical stretch-induced human endothelial cell reorientation and convergent extension cell movement in zebrafish gastrula. In this study, we show that Solo binds to keratin-8/keratin-18 (K8/K18) intermediate filaments through multiple sites. Solo overexpression promotes the formation of thick actin stress fibers and keratin bundles, whereas knockdown of Solo, expression of a GEF-inactive mutant of Solo, or inhibition of ROCK suppresses stress fiber formation and leads to disorganized keratin networks, indicating that the Solo-RhoA-ROCK pathway serves to precisely organize keratin networks, as well as to promote stress fibers. Of importance, knockdown of Solo or K18 or overexpression of GEF-inactive or deletion mutants of Solo suppresses tensile force-induced stress fiber reinforcement. Furthermore, knockdown of Solo or K18 suppresses tensile force-induced RhoA activation. These results strongly suggest that the interplay between Solo and K8/K18 filaments plays a crucial role in tensile force-induced RhoA activation and consequent actin cytoskeletal reinforcement. © 2016 Fujiwara et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

National Centers for Environmental Prediction

Science.gov Websites

Statistics Observational Data Processing Data Assimilation Monsoon Desk Model Transition Seminars Seminar (CFS) HURRICANE WEATHER RESEARCH and FORECASTING (HWRF) GLOBAL ENSEMBLE FORECAST SYSTEM (GEFS) NATIONAL Climate Prediction (NCWCP) 5830 University Research Court College Park, MD 20740 Page Author: EMC

ARF6 Activated by the LHCG Receptor through the Cytohesin Family of Guanine Nucleotide Exchange Factors Mediates the Receptor Internalization and Signaling*

PubMed Central

Kanamarlapudi, Venkateswarlu; Thompson, Aiysha; Kelly, Eamonn; López Bernal, Andrés

2012-01-01

The luteinizing hormone chorionic gonadotropin receptor (LHCGR) is a Gs-coupled GPCR that is essential for the maturation and function of the ovary and testis. LHCGR is internalized following its activation, which regulates the biological responsiveness of the receptor. Previous studies indicated that ADP-ribosylation factor (ARF)6 and its GTP-exchange factor (GEF) cytohesin 2 regulate LHCGR internalization in follicular membranes. However, the mechanisms by which ARF6 and cytohesin 2 regulate LHCGR internalization remain incompletely understood. Here we investigated the role of the ARF6 signaling pathway in the internalization of heterologously expressed human LHCGR (HLHCGR) in intact cells using a combination of pharmacological inhibitors, siRNA and the expression of mutant proteins. We found that human CG (HCG)-induced HLHCGR internalization, cAMP accumulation and ARF6 activation were inhibited by Gallein (βγ inhibitor), Wortmannin (PI 3-kinase inhibitor), SecinH3 (cytohesin ARF GEF inhibitor), QS11 (an ARF GAP inhibitor), an ARF6 inhibitory peptide and ARF6 siRNA. However, Dynasore (dynamin inhibitor), the dominant negative mutants of NM23-H1 (dynamin activator) and clathrin, and PBP10 (PtdIns 4,5-P2-binding peptide) inhibited agonist-induced HLHCGR and cAMP accumulation but not ARF6 activation. These results indicate that heterotrimeric G-protein, phosphatidylinositol (PI) 3-kinase (PI3K), cytohesin ARF GEF and ARF GAP function upstream of ARF6 whereas dynamin and clathrin act downstream of ARF6 in the regulation of HCG-induced HLHCGR internalization and signaling. In conclusion, we have identified the components and molecular details of the ARF6 signaling pathway required for agonist-induced HLHCGR internalization. PMID:22523074

Huperzine A Provides Robust and Sustained Protection against Induced Seizures in Scn1a Mutant Mice

PubMed Central

Wong, Jennifer C.; Dutton, Stacey B. B.; Collins, Stephen D.; Schachter, Steven; Escayg, Andrew

2016-01-01

De novo loss-of-function mutations in the voltage-gated sodium channel (VGSC) SCN1A (encoding Nav1.1) are the main cause of Dravet syndrome (DS), a catastrophic early-life encephalopathy associated with prolonged and recurrent early-life febrile seizures (FSs), refractory afebrile epilepsy, cognitive and behavioral deficits, and a 15–20% mortality rate. SCN1A mutations also lead to genetic epilepsy with febrile seizures plus (GEFS+), which is an inherited disorder characterized by early-life FSs and the development of a range of adult epilepsy subtypes. Current antiepileptic drugs often fail to protect against the severe seizures and behavioral and cognitive deficits found in patients with SCN1A mutations. To address the need for more efficacious treatments for SCN1A-derived epilepsies, we evaluated the therapeutic potential of Huperzine A, a naturally occurring reversible acetylcholinesterase inhibitor. In CF1 mice, Hup A (0.56 or 1 mg/kg) was found to confer protection against 6 Hz-, pentylenetetrazole (PTZ)-, and maximal electroshock (MES)-induced seizures. Robust protection against 6 Hz-, MES-, and hyperthermia-induced seizures was also achieved following Hup A administration in mouse models of DS (Scn1a+/−) and GEFS+ (Scn1aRH/+). Furthermore, Hup A-mediated seizure protection was sustained during 3 weeks of daily injections in Scn1aRH/+ mutants. Finally, we determined that muscarinic and GABAA receptors play a role in Hup A-mediated seizure protection. These findings indicate that Hup A might provide a novel therapeutic strategy for increasing seizure resistance in DS and GEFS+, and more broadly, in other forms of refractory epilepsy. PMID:27799911

MJO simulation in CMIP5 climate models: MJO skill metrics and process-oriented diagnosis

NASA Astrophysics Data System (ADS)

Ahn, Min-Seop; Kim, Daehyun; Sperber, Kenneth R.; Kang, In-Sik; Maloney, Eric; Waliser, Duane; Hendon, Harry

2017-12-01

The Madden-Julian Oscillation (MJO) simulation diagnostics developed by MJO Working Group and the process-oriented MJO simulation diagnostics developed by MJO Task Force are applied to 37 Coupled Model Intercomparison Project phase 5 (CMIP5) models in order to assess model skill in representing amplitude, period, and coherent eastward propagation of the MJO, and to establish a link between MJO simulation skill and parameterized physical processes. Process-oriented diagnostics include the Relative Humidity Composite based on Precipitation (RHCP), Normalized Gross Moist Stability (NGMS), and the Greenhouse Enhancement Factor (GEF). Numerous scalar metrics are developed to quantify the results. Most CMIP5 models underestimate MJO amplitude, especially when outgoing longwave radiation (OLR) is used in the evaluation, and exhibit too fast phase speed while lacking coherence between eastward propagation of precipitation/convection and the wind field. The RHCP-metric, indicative of the sensitivity of simulated convection to low-level environmental moisture, and the NGMS-metric, indicative of the efficiency of a convective atmosphere for exporting moist static energy out of the column, show robust correlations with a large number of MJO skill metrics. The GEF-metric, indicative of the strength of the column-integrated longwave radiative heating due to cloud-radiation interaction, is also correlated with the MJO skill metrics, but shows relatively lower correlations compared to the RHCP- and NGMS-metrics. Our results suggest that modifications to processes associated with moisture-convection coupling and the gross moist stability might be the most fruitful for improving simulations of the MJO. Though the GEF-metric exhibits lower correlations with the MJO skill metrics, the longwave radiation feedback is highly relevant for simulating the weak precipitation anomaly regime that may be important for the establishment of shallow convection and the transition to deep convection.

MJO simulation in CMIP5 climate models: MJO skill metrics and process-oriented diagnosis

DOE PAGES

Ahn, Min-Seop; Kim, Daehyun; Sperber, Kenneth R.; ...

2017-03-23

The Madden-Julian Oscillation (MJO) simulation diagnostics developed by MJO Working Group and the process-oriented MJO simulation diagnostics developed by MJO Task Force are applied to 37 Coupled Model Intercomparison Project phase 5 (CMIP5) models in order to assess model skill in representing amplitude, period, and coherent eastward propagation of the MJO, and to establish a link between MJO simulation skill and parameterized physical processes. Process-oriented diagnostics include the Relative Humidity Composite based on Precipitation (RHCP), Normalized Gross Moist Stability (NGMS), and the Greenhouse Enhancement Factor (GEF). Numerous scalar metrics are developed to quantify the results. Most CMIP5 models underestimate MJOmore » amplitude, especially when outgoing longwave radiation (OLR) is used in the evaluation, and exhibit too fast phase speed while lacking coherence between eastward propagation of precipitation/convection and the wind field. The RHCP-metric, indicative of the sensitivity of simulated convection to low-level environmental moisture, and the NGMS-metric, indicative of the efficiency of a convective atmosphere for exporting moist static energy out of the column, show robust correlations with a large number of MJO skill metrics. The GEF-metric, indicative of the strength of the column-integrated longwave radiative heating due to cloud-radiation interaction, is also correlated with the MJO skill metrics, but shows relatively lower correlations compared to the RHCP- and NGMS-metrics. Our results suggest that modifications to processes associated with moisture-convection coupling and the gross moist stability might be the most fruitful for improving simulations of the MJO. Though the GEF-metric exhibits lower correlations with the MJO skill metrics, the longwave radiation feedback is highly relevant for simulating the weak precipitation anomaly regime that may be important for the establishment of shallow convection and the transition to deep convection.« less

MJO simulation in CMIP5 climate models: MJO skill metrics and process-oriented diagnosis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ahn, Min-Seop; Kim, Daehyun; Sperber, Kenneth R.

The Madden-Julian Oscillation (MJO) simulation diagnostics developed by MJO Working Group and the process-oriented MJO simulation diagnostics developed by MJO Task Force are applied to 37 Coupled Model Intercomparison Project phase 5 (CMIP5) models in order to assess model skill in representing amplitude, period, and coherent eastward propagation of the MJO, and to establish a link between MJO simulation skill and parameterized physical processes. Process-oriented diagnostics include the Relative Humidity Composite based on Precipitation (RHCP), Normalized Gross Moist Stability (NGMS), and the Greenhouse Enhancement Factor (GEF). Numerous scalar metrics are developed to quantify the results. Most CMIP5 models underestimate MJOmore » amplitude, especially when outgoing longwave radiation (OLR) is used in the evaluation, and exhibit too fast phase speed while lacking coherence between eastward propagation of precipitation/convection and the wind field. The RHCP-metric, indicative of the sensitivity of simulated convection to low-level environmental moisture, and the NGMS-metric, indicative of the efficiency of a convective atmosphere for exporting moist static energy out of the column, show robust correlations with a large number of MJO skill metrics. The GEF-metric, indicative of the strength of the column-integrated longwave radiative heating due to cloud-radiation interaction, is also correlated with the MJO skill metrics, but shows relatively lower correlations compared to the RHCP- and NGMS-metrics. Our results suggest that modifications to processes associated with moisture-convection coupling and the gross moist stability might be the most fruitful for improving simulations of the MJO. Though the GEF-metric exhibits lower correlations with the MJO skill metrics, the longwave radiation feedback is highly relevant for simulating the weak precipitation anomaly regime that may be important for the establishment of shallow convection and the transition to deep convection.« less

The N Termini of a-Subunit Isoforms Are Involved in Signaling between Vacuolar H+-ATPase (V-ATPase) and Cytohesin-2*

PubMed Central

Hosokawa, Hiroyuki; Dip, Phat Vinh; Merkulova, Maria; Bakulina, Anastasia; Zhuang, Zhenjie; Khatri, Ashok; Jian, Xiaoying; Keating, Shawn M.; Bueler, Stephanie A.; Rubinstein, John L.; Randazzo, Paul A.; Ausiello, Dennis A.; Grüber, Gerhard; Marshansky, Vladimir

2013-01-01

Previously, we reported an acidification-dependent interaction of the endosomal vacuolar H+-ATPase (V-ATPase) with cytohesin-2, a GDP/GTP exchange factor (GEF), suggesting that it functions as a pH-sensing receptor. Here, we have studied the molecular mechanism of signaling between the V-ATPase, cytohesin-2, and Arf GTP-binding proteins. We found that part of the N-terminal cytosolic tail of the V-ATPase a2-subunit (a2N), corresponding to its first 17 amino acids (a2N(1–17)), potently modulates the enzymatic GDP/GTP exchange activity of cytohesin-2. Moreover, this peptide strongly inhibits GEF activity via direct interaction with the Sec7 domain of cytohesin-2. The structure of a2N(1–17) and its amino acids Phe5, Met10, and Gln14 involved in interaction with Sec7 domain were determined by NMR spectroscopy analysis. In silico docking experiments revealed that part of the V-ATPase formed by its a2N(1–17) epitope competes with the switch 2 region of Arf1 and Arf6 for binding to the Sec7 domain of cytohesin-2. The amino acid sequence alignment and GEF activity studies also uncovered the conserved character of signaling between all four (a1–a4) a-subunit isoforms of mammalian V-ATPase and cytohesin-2. Moreover, the conserved character of this phenomenon was also confirmed in experiments showing binding of mammalian cytohesin-2 to the intact yeast V-ATPase holo-complex. Thus, here we have uncovered an evolutionarily conserved function of the V-ATPase as a novel cytohesin-signaling receptor. PMID:23288846

National Centers for Environmental Prediction

Science.gov Websites

/ VISION | About EMC EMC > GEFS > COLLABORATORS Home Operational Products Experimental Data ENSEMBLE FORECAST SYSTEM MSC NAEFS Products CPC NAEFS Experimental 8 to 14 Day Temperature Guidance CPC NAEFS Experimental 8 to 14 Day Precip Guidance NOAA / National Weather Service National Centers for

Gain-of-function SOS1 mutations cause a distinctive form of noonansyndrome

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tartaglia, Marco; Pennacchio, Len A.; Zhao, Chen

2006-09-01

Noonan syndrome (NS) is a developmental disordercharacterized by short stature, facial dysmorphia, congenital heartdefects and skeletal anomalies1. Increased RAS-mitogenactivated proteinkinase (MAPK) signaling due to PTPN11 and KRAS mutations cause 50 percentof NS2-6. Here, we report that 22 of 129 NS patients without PTPN11 orKRAS mutation (17 percent) have missense mutations in SOS1, which encodesa RAS-specific guanine nucleotide exchange factor (GEF). SOS1 mutationscluster at residues implicated in the maintenance of SOS1 in itsautoinhibited form and ectopic expression of two NS-associated mutantsinduced enhanced RAS activation. The phenotype associated with SOS1defects is distinctive, although within NS spectrum, with a highprevalence of ectodermal abnormalitiesmore » but generally normal developmentand linear growth. Our findings implicate for the first timegain-of-function mutations in a RAS GEF in inherited disease and define anew mechanism by which upregulation of the RAS pathway can profoundlychange human development.« less

Gridded Calibration of Ensemble Wind Vector Forecasts Using Ensemble Model Output Statistics

NASA Astrophysics Data System (ADS)

Lazarus, S. M.; Holman, B. P.; Splitt, M. E.

2017-12-01

A computationally efficient method is developed that performs gridded post processing of ensemble wind vector forecasts. An expansive set of idealized WRF model simulations are generated to provide physically consistent high resolution winds over a coastal domain characterized by an intricate land / water mask. Ensemble model output statistics (EMOS) is used to calibrate the ensemble wind vector forecasts at observation locations. The local EMOS predictive parameters (mean and variance) are then spread throughout the grid utilizing flow-dependent statistical relationships extracted from the downscaled WRF winds. Using data withdrawal and 28 east central Florida stations, the method is applied to one year of 24 h wind forecasts from the Global Ensemble Forecast System (GEFS). Compared to the raw GEFS, the approach improves both the deterministic and probabilistic forecast skill. Analysis of multivariate rank histograms indicate the post processed forecasts are calibrated. Two downscaling case studies are presented, a quiescent easterly flow event and a frontal passage. Strengths and weaknesses of the approach are presented and discussed.

Multiple roles of the Rho GEF ephexin1 in synapse remodeling

PubMed Central

Shi, Lei; Fu, Amy KY

2010-01-01

Synapse remodeling, which involves changes in the synaptic structure and their molecular composition, is required for the maturation and refinement of neural circuits. Although synapse remodeling is known to be tightly dependent on the assembly of local actin cytoskeleton, how actin directs the structural changes of synapse and targeting of synaptic proteins are not fully understood. Recently, we identified ephexin1, a Rho guanine nucleotide exchange factor (GEF) that regulates actin dynamics, to play an essential role in the maturation and functioning of the mammalian neuromuscular junction (NMJ). We showed that ephexin1 regulates the synaptic organization of the neurotransmitter receptor acetylcholine receptor (AChR) clusters through RhoA-dependent actin reorganization. Interestingly, ephexin1 has been implicated in the regulation of postsynaptic structure as well as the presynaptic vesicle release at various types of synapses. Our findings thus establish a novel function of ephexin1 in synapse remodeling through regulating the synaptic targeting of neurotransmitter receptors, revealing a versatile role of ephexin1 at synapses. PMID:21331259

Mechanism of SOS PR-domain autoinhibition revealed by single-molecule assays on native protein from lysate

PubMed Central

Lee, Young Kwang; Low-Nam, Shalini T.; Chung, Jean K.; Hansen, Scott D.; Lam, Hiu Yue Monatrice; Alvarez, Steven; Groves, Jay T.

2017-01-01

The guanine nucleotide exchange factor (GEF) Son of Sevenless (SOS) plays a critical role in signal transduction by activating Ras. Here we introduce a single-molecule assay in which individual SOS molecules are captured from raw cell lysate using Ras-functionalized supported membrane microarrays. This enables characterization of the full-length SOS protein, which has not previously been studied in reconstitution due to difficulties in purification. Our measurements on the full-length protein reveal a distinct role of the C-terminal proline-rich (PR) domain to obstruct the engagement of allosteric Ras independently of the well-known N-terminal domain autoinhibition. This inhibitory role of the PR domain limits Grb2-independent recruitment of SOS to the membrane through binding of Ras·GTP in the SOS allosteric binding site. More generally, this assay strategy enables characterization of the functional behaviour of GEFs with single-molecule precision but without the need for purification. PMID:28452363

Mechanism of SOS PR-domain autoinhibition revealed by single-molecule assays on native protein from lysate.

PubMed

Lee, Young Kwang; Low-Nam, Shalini T; Chung, Jean K; Hansen, Scott D; Lam, Hiu Yue Monatrice; Alvarez, Steven; Groves, Jay T

2017-04-28

The guanine nucleotide exchange factor (GEF) Son of Sevenless (SOS) plays a critical role in signal transduction by activating Ras. Here we introduce a single-molecule assay in which individual SOS molecules are captured from raw cell lysate using Ras-functionalized supported membrane microarrays. This enables characterization of the full-length SOS protein, which has not previously been studied in reconstitution due to difficulties in purification. Our measurements on the full-length protein reveal a distinct role of the C-terminal proline-rich (PR) domain to obstruct the engagement of allosteric Ras independently of the well-known N-terminal domain autoinhibition. This inhibitory role of the PR domain limits Grb2-independent recruitment of SOS to the membrane through binding of Ras·GTP in the SOS allosteric binding site. More generally, this assay strategy enables characterization of the functional behaviour of GEFs with single-molecule precision but without the need for purification.

Developmental lineage priming in Dictyostelium by heterogeneous Ras activation.

PubMed

Chattwood, Alex; Nagayama, Koki; Bolourani, Parvin; Harkin, Lauren; Kamjoo, Marzieh; Weeks, Gerald; Thompson, Christopher R L

2013-11-26

In cell culture, genetically identical cells often exhibit heterogeneous behavior, with only 'lineage primed' cells responding to differentiation inducing signals. It has recently been proposed that such heterogeneity exists during normal embryonic development to allow position independent patterning based on 'salt and pepper' differentiation and sorting out. However, the molecular basis of lineage priming and how it leads to reproducible cell type proportioning are poorly understood. To address this, we employed a novel forward genetic approach in the model organism Dictyostelium discoideum. These studies reveal that the Ras-GTPase regulator gefE is required for normal lineage priming and salt and pepper differentiation. This is because Ras-GTPase activity sets the intrinsic response threshold to lineage specific differentiation signals. Importantly, we show that although gefE expression is uniform, transcription of its target, rasD, is both heterogeneous and dynamic, thus providing a novel mechanism for heterogeneity generation and position-independent differentiation. DOI: http://dx.doi.org/10.7554/eLife.01067.001.

Neutron-induced fission cross-section measurement of 234U with quasi-monoenergetic beams in the keV and MeV range using micromegas detectors

NASA Astrophysics Data System (ADS)

Tsinganis, A.; Kokkoris, M.; Vlastou, R.; Kalamara, A.; Stamatopoulos, A.; Kanellakopoulos, A.; Lagoyannis, A.; Axiotis, M.

2017-09-01

Accurate data on neutron-induced fission cross-sections of actinides are essential for the design of advanced nuclear reactors based either on fast neutron spectra or alternative fuel cycles, as well as for the reduction of safety margins of existing and future conventional facilities. The fission cross-section of 234U was measured at incident neutron energies of 560 and 660 keV and 7.5 MeV with a setup based on `microbulk' Micromegas detectors and the same samples previously used for the measurement performed at the CERN n_TOF facility (Karadimos et al., 2014). The 235U fission cross-section was used as reference. The (quasi-)monoenergetic neutron beams were produced via the 7Li(p,n) and the 2H(d,n) reactions at the neutron beam facility of the Institute of Nuclear and Particle Physics at the `Demokritos' National Centre for Scientific Research. A detailed study of the neutron spectra produced in the targets and intercepted by the samples was performed coupling the NeuSDesc and MCNPX codes, taking into account the energy spread, energy loss and angular straggling of the beam ions in the target assemblies, as well as contributions from competing reactions and neutron scattering in the experimental setup. Auxiliary Monte-Carlo simulations were performed with the FLUKA code to study the behaviour of the detectors, focusing particularly on the reproduction of the pulse height spectra of α-particles and fission fragments (using distributions produced with the GEF code) for the evaluation of the detector efficiency. An overview of the developed methodology and preliminary results are presented.

RAF and Protection Warfighting Function

DTIC Science & Technology

2015-02-01

countries and challenges (including both the Asian Pacific and the Middle East).10 To execute this strategy in an increasingly complex world, the...of the Force (GEF) to prepare and review theatre campaign plans (TCP).27 The TCP is the CCDR’s vehicle for operationalizing the theater strategy. The

NCEP Data Products

Science.gov Websites

Image of NCEP Logo WHERE AMERICA'S CLIMATE AND WEATHER SERVICES BEGIN Inventory of Data Products on Generated Products Image of horizontal rule Global Forecast System (GFS) GFS Ensemble Forecast System (GEFS of horizontal rule External Products Image of horizontal rule Canadian Ensemble Forecast System

Extratropical Cyclones Leading to Extreme Weather Events over Central and Eastern North America

NASA Astrophysics Data System (ADS)

Bentley, Alicia M.

Cool-season extreme weather events (EWEs) occurring over central and eastern North America are typically associated with strong extratropical cyclones (ECs) that are governed by varying combinations of baroclinic, diabatic, and barotropic processes. This dissertation investigates the climatology, evolution, and predictability of ECs leading to EWEs over central and eastern North America, and provides a foundation on which to compare ECs leading to EWEs to ordinary ECs forming over and traversing the same regions. A climatology of ECs leading to EWEs over central and eastern North America during October-March 1979-2016 reveals that these ECs typically form 1) in the lee of the Rocky Mountains, 2) over the south central U.S., and 3) along the east coast of North America at latitudes equatorward of the typical genesis locations of ordinary ECs. ECs leading to EWEs included in the climatology form most frequently in November and March, when the seasonal alignment of baroclinic and convectively driven forcings occurs. Consistent with previous studies of North American ECs, the location and frequency of ECs leading to EWEs are partially determined by the states of the Pacific-North American pattern and the North Atlantic Oscillation. Metrics representing baroclinic, diabatic, and barotropic processes are formulated in this dissertation and are used to determine the combinations of baroclinic, diabatic, and barotropic processes associated with the formation and maintenance of ordinary ECs and ECs leading to EWEs. These metrics reveal that ECs leading to EWEs are associated with contributions from baroclinic, diabatic, and barotropic processes that are 1) similar to those associated with ordinary ECs at the time of formation (t0) and 2) considerably larger than those associated with ordinary ECs at the time of maximum intensity (tmax). Baroclinic processes typically contribute more than diabatic and barotropic processes throughout the evolution of ECs leading to EWEs. Diabatic processes typically contribute more during the intensification of ECs leading to EWEs than during their maintenance after tmax, whereas barotropic processes typically contribute more during the maintenance of ECs leading to EWEs after tmax than during their intensification. The relative contributions from baroclinic, diabatic, and barotropic processes during the evolution of ECs leading to EWEs are also shown to differ based on their genesis location. The 1.0° NOAA Global Ensemble Forecast System (GEFS) reforecast dataset is used in this dissertation to evaluate the forecast skill associated with ordinary ECs and ECs leading to EWEs at 0-192-h lead times. Ordinary ECs are consistently too slow and left of track in the GEFS, and are often too weak at longer lead times. ECs leading to EWEs are consistently too weak, fast, and right of track in the GEFS at longer lead times, and consistently too strong, slow, and left of track at shorter lead times. The positions of ordinary ECs are forecast with less skill and more spread than the positions of ECs leading to EWEs in the GEFS, whereas the intensities of ordinary ECs and ECs leading to EWEs are forecast with similar skill and spread. Locations over central and eastern North America where the positions and intensities of ordinary ECs and ECs leading to EWEs are frequently forecast with relatively low and high skill and spread in the GEFS are also identified.

Towards integrated solutions for water, energy, and land using an integrated nexus modeling framework

NASA Astrophysics Data System (ADS)

Wada, Y.

2017-12-01

Humanity has already reached or even exceeded the Earth's carrying capacity. Growing needs for food, energy and water will only exacerbate existing challenges over the next decades. Consequently, the acceptance of "business as usual" is eroding and we are being challenged to adopt new, more integrated, and more inclusive development pathways that avoid dangerous interference with the local environment and global planetary boundaries. This challenge is embodied in the United Nation's Sustainable Development Goals (SDGs), which endeavor to set a global agenda for moving towards more sustainable development strategies. To improve and sustain human welfare, it is critical that access to modern, reliable, and affordable water, energy, and food is expanded and maintained. The Integrated Solutions for Water, Energy, and Land (IS-WEL) project has been launched by IIASA, together with the Global Environment Facility (GEF) and the United Nations Industrial Development Organization (UNIDO). This project focuses on the water-energy-land nexus in the context of other major global challenges such as urbanization, environmental degradation, and equitable and sustainable futures. It develops a consistent framework for looking at the water-energy-land nexus and identify strategies for achieving the needed transformational outcomes through an advanced assessment framework. A multi-scalar approach are being developed that aims to combine global and regional integrated assessment tools with local stakeholder knowledge in order to identify robust solutions to energy, water, food, and ecosystem security in selected regions of the world. These are regions facing multiple energy, water and land use challenges and rapid demographic and economic changes, and are hardest hit by increasing climate variability and change. This project combines the global integrated assessment model (MESSAGE) with the global land (GLOBIOM) and water (Community Water Model) model respectively, and the integrated modeling framework are then combined with detailed regional decision support tools for water-energy-land nexus analysis in case study regions. A number of stakeholder meetings are used to engage local communities in the definition of important nexus drivers, scenario development and definition of performance metrics.

NAEFS-2007

Science.gov Websites

the NCO PMB model changes web site for more information concerning NCEP model changes. http ://www.nco.ncep.noaa.gov/pmb/changes/ Thanks, Joey Carr EFFECTIVE TUE DEC 4 2007...WITH THE 1200 COORDINATED UNIVERSAL TIME /CONTENT CHANGES: NOAAPORT DATA: - THE NCEP GEFS ENSEMBLE PRODUCTS WHICH ARE DELIVERED TO NOAAPORT

The Juxtamembrane and carboxy-terminal domains of Arabidopsis PRK2 are critical for ROP-induced growth in pollen tubes

USDA-ARS?s Scientific Manuscript database

Polarized growth of pollen tubes is a critical step for successful reproduction in angiosperms and is controlled by ROP GTPases. Spatiotemporal activation of ROP (Rho GTPases of plants) necessitates a complex and sophisticated regulatory system, in which guanine nucleotide exchange factors (RopGEFs)...

Airfield Damage Recovery Techniques of 18th Engineer Brigade in Europe

DTIC Science & Technology

1980-12-01

have to be observed. Comparable German regulations are published in "Verordnung uber gefAbrliche Arbeitsstoffe", issue May 1, 1976. Ŗ.1. Most important...rules: Rubber boots, acid-prove protective gloves, protective facial masks, water spray bottles for eyes 2.2. No smoking, no eating at jobside 3

Structural Dynamics Control Allosteric Activation of Cytohesin Family Arf GTPase Exchange Factors

DOE Office of Scientific and Technical Information (OSTI.GOV)

Malaby, Andrew W.; Das, Sanchaita; Chakravarthy, Srinivas

Membrane dynamic processes including vesicle biogenesis depend on Arf guanosine triphosphatase (GTPase) activation by guanine nucleotide exchange factors (GEFs) containing a catalytic Sec7 domain and a membrane-targeting module such as a pleckstrin homology (PH) domain. The catalytic output of cytohesin family Arf GEFs is controlled by autoinhibitory interactions that impede accessibility of the exchange site in the Sec7 domain. These restraints can be relieved through activator Arf-GTP binding to an allosteric site comprising the PH domain and proximal autoinhibitory elements (Sec7-PH linker and C-terminal helix). Small-angle X-ray scattering and negative-stain electron microscopy were used to investigate the structural organization andmore » conformational dynamics of cytohesin-3 (Grp1) in autoinhibited and active states. The results support a model in which hinge dynamics in the autoinhibited state expose the activator site for Arf-GTP binding, while subsequent C-terminal helix unlatching and repositioning unleash conformational entropy in the Sec7-PH linker to drive exposure of the exchange site.« less

Targeted Sos1 deletion reveals its critical role in early T-cell development

PubMed Central

Kortum, Robert L.; Sommers, Connie L.; Alexander, Clayton P.; Pinski, John M.; Li, Wenmei; Grinberg, Alex; Lee, Jan; Love, Paul E.; Samelson, Lawrence E.

2011-01-01

Activation of the small G protein Ras is required for thymocyte differentiation. In thymocytes, Ras is activated by the Ras guanine exchange factors (RasGEFs) Sos1, Sos2, and RasGRP1. We report the development of a floxed allele of sos1 to assess the role of Sos1 during thymocyte development. Sos1 was required for pre–T-cell receptor (pre-TCR)– but not TCR-stimulated developmental signals. Sos1 deletion led to a partial block at the DN-to-DP transition. Sos1-deficient thymocytes showed reduced pre-TCR–stimulated proliferation, differentiation, and ERK phosphorylation. In contrast, TCR-stimulated positive selection, and negative selection under strong stimulatory conditions, remained intact in Sos1-deficient mice. Comparison of RasGEF expression at different developmental stages showed that relative to Sos2 and RasGRP1, Sos1 is most abundant in DN thymocytes, but least abundant in DP thymocytes. These data reveal that Sos1 is uniquely positioned to affect signal transduction early in thymocyte development. PMID:21746917

Identification of a negative regulatory region for the exchange activity and characterization of T332I mutant of Rho guanine nucleotide exchange factor 10 (ARHGEF10).

PubMed

Chaya, Taro; Shibata, Satoshi; Tokuhara, Yasunori; Yamaguchi, Wataru; Matsumoto, Hiroshi; Kawahara, Ichiro; Kogo, Mikihiko; Ohoka, Yoshiharu; Inagaki, Shinobu

2011-08-26

The T332I mutation in Rho guanine nucleotide exchange factor 10 (ARHGEF10) was previously found in persons with slowed nerve conduction velocities and thin myelination of peripheral nerves. However, the molecular and cellular basis of the T332I mutant is not understood. Here, we show that ARHGEF10 has a negative regulatory region in the N terminus, in which residue 332 is located, and the T332I mutant is constitutively active. An N-terminal truncated ARHGEF10 mutant, ARHGEF10 ΔN (lacking amino acids 1-332), induced cell contraction that was inhibited by a Rho kinase inhibitor Y27632 and had higher GEF activity for RhoA than the wild type. The T332I mutant also showed the phenotype similar to the N-terminal truncated mutant. These data suggest that the ARHGEF10 T332I mutation-associated phenotype observed in the peripheral nerves is due to activated GEF activity of the ARHGEF10 T332I mutant.

Identification of a Negative Regulatory Region for the Exchange Activity and Characterization of T332I Mutant of Rho Guanine Nucleotide Exchange Factor 10 (ARHGEF10)*

PubMed Central

Chaya, Taro; Shibata, Satoshi; Tokuhara, Yasunori; Yamaguchi, Wataru; Matsumoto, Hiroshi; Kawahara, Ichiro; Kogo, Mikihiko; Ohoka, Yoshiharu; Inagaki, Shinobu

2011-01-01

The T332I mutation in Rho guanine nucleotide exchange factor 10 (ARHGEF10) was previously found in persons with slowed nerve conduction velocities and thin myelination of peripheral nerves. However, the molecular and cellular basis of the T332I mutant is not understood. Here, we show that ARHGEF10 has a negative regulatory region in the N terminus, in which residue 332 is located, and the T332I mutant is constitutively active. An N-terminal truncated ARHGEF10 mutant, ARHGEF10 ΔN (lacking amino acids 1–332), induced cell contraction that was inhibited by a Rho kinase inhibitor Y27632 and had higher GEF activity for RhoA than the wild type. The T332I mutant also showed the phenotype similar to the N-terminal truncated mutant. These data suggest that the ARHGEF10 T332I mutation-associated phenotype observed in the peripheral nerves is due to activated GEF activity of the ARHGEF10 T332I mutant. PMID:21719701

The product of C9orf72, a gene strongly implicated in neurodegeneration, is structurally related to DENN Rab-GEFs.

PubMed

Levine, Timothy P; Daniels, Rachel D; Gatta, Alberto T; Wong, Louise H; Hayes, Matthew J

2013-02-15

Fronto-temporal dementia (FTD) and amyotrophic lateral sclerosis (ALS, also called motor neuron disease, MND) are severe neurodegenerative diseases that show considerable overlap at the clinical and cellular level. The most common single mutation in families with FTD or ALS has recently been mapped to a non-coding repeat expansion in the uncharacterized gene C9ORF72. Although a plausible mechanism for disease is that aberrant C9ORF72 mRNA poisons splicing, it is important to determine the cellular function of C9ORF72, about which nothing is known. Sensitive homology searches showed that C9ORF72 is a full-length distant homologue of proteins related to Differentially Expressed in Normal and Neoplasia (DENN), which is a GDP/GTP exchange factor (GEF) that activates Rab-GTPases. Our results suggest that C9ORF72 is likely to regulate membrane traffic in conjunction with Rab-GTPase switches, and we propose to name the gene and its product DENN-like 72 (DENNL72).

Reduced cell number in the hindgut epithelium disrupts hindgut left-right asymmetry in a mutant of pebble, encoding a RhoGEF, in Drosophila embryos.

PubMed

Nakamura, Mitsutoshi; Matsumoto, Kenjiroo; Iwamoto, Yuta; Muguruma, Takeshi; Nakazawa, Naotaka; Hatori, Ryo; Taniguchi, Kiichiro; Maeda, Reo; Matsuno, Kenji

2013-02-01

Animals often show left-right (LR) asymmetry in their body structures. In some vertebrates, the mechanisms underlying LR symmetry breaking and the subsequent signals responsible for LR asymmetric development are well understood. However, in invertebrates, the molecular bases of these processes are largely unknown. Therefore, we have been studying the genetic pathway of LR asymmetric development in Drosophila. The embryonic gut is the first organ that shows directional LR asymmetry during Drosophila development. We performed a genetic screen to identify mutations affecting LR asymmetric development of the embryonic gut. From this screen, we isolated pebble (pbl), which encodes a homolog of a mammalian RhoGEF, Ect2. The laterality of the hindgut was randomized in embryos homozygous for a null mutant of pbl. Pbl is a multi-functional protein required for cytokinesis and the epithelial-to-mesenchymal transition in Drosophila. Consistent with Pbl's role in cytokinesis, we found reduced numbers of cells in the hindgut epithelium in pbl homozygous embryos. The specific expression of pbl in the hindgut epithelium, but not in other tissues, rescued the LR defects and reduced cell number in embryonic pbl homozygotes. Embryos homozygous for string (stg), a mutant that reduces cell number through a different mechanism, also showed LR defects of the hindgut. However, the reduction in cell number in the pbl mutants was not accompanied by defects in the specification of hindgut epithelial tissues or their integrity. Based on these results, we speculate that the reduction in cell number may be one reason for the LR asymmetry defect of the pbl hindgut, although we cannot exclude contributions from other functions of Pbl, including regulation of the actin cytoskeleton through its RhoGEF activity. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

[Virulence determinant of Chromobacterium violaceum].

PubMed

Miki, Tsuyoshi

2014-01-01

Chromobacterium violaceum is a Gram-negative bacterium that infects humans and animals with fatal sepsis. The infection with C. violaceum is rare in case of those who are healthy, but once established, C. violaceum causes sever disease accompanied by abscess formation in the lungs, liver and spleen. Furthermore, C. violaceum is resistant to a broad range of antibiotics, which in some cases renders the antimicrobial therapy for this infection difficult. Thus, the infection with C. violaceum displays high mortality rates unless initial proper antimicrobial therapy. In contrast, the infection mechanism had completely remained unknown. To this end, we have tried to identify virulence factors-associated with C. violaceum infection. Two distinct type III secretion systems (TTSSs) were thought to be one of the most important virulence factors, which are encoded by Chromobacterium pathogenicity island 1/1a and 2 (Cpi-1/-1a and -2) respectively. Our results have shown that Cpi-1/-1a-encoded TTSS, but not Cpi-2, is indispensable for the virulence in a mouse infection model. C. violaceum caused fulminant hepatitis in a Cpi-1/-1a-encoded TTSS-dependent manner. We next have identified 16 novel effectors secreted from Cpi-1/-1a-encoded TTS machinery. From these effectors, we found that CopE (Chromobacterium outer protein E) has similarities to a guanine nucleotide exchange factor (GEF) for Rho GTPases. CopE acts as GEF for Rac1 and Cdc42, leading to induction of actin cytoskeletal rearrangement. Interestingly, C. violaceum invades cultured human epithelial cells in a CopE-dependent manner. Finally, an inactivation of CopE by disruption of copE gene or amino acid point mutation leading to loss of GEF activity attenuates significantly the mouse virulence of C. violaceum. These results suggest that Cpi-1/-1a-encoded TTSS is a major virulence determinant for C. violaceum infection, and that CopE contributes to the virulence in part of this pathogen.

Cdc42 and the Guanine Nucleotide Exchange Factors Ect2 and Trio Mediate Fn14-Induced Migration and Invasion of Glioblastoma Cells

PubMed Central

Fortin, Shannon P.; Ennis, Matthew J.; Schumacher, Cassie A.; Zylstra-Diegel, Cassandra R.; Williams, Bart O.; Ross, Julianna T.D.; Winkles, Jeffrey A.; Loftus, Joseph C.; Symons, Marc H.; Tran, Nhan L.

2012-01-01

Malignant glioblastomas are characterized by their ability to infiltrate into normal brain. We previously reported that binding of the multifunctional cytokine TNF-like weak inducer of apoptosis (TWEAK) to its receptor fibroblast growth factor–inducible 14 (Fn14) induces glioblastoma cell invasion via Rac1 activation. Here, we show that Cdc42 plays an essential role in Fn14-mediated activation of Rac1. TWEAK-treated glioma cells display an increased activation of Cdc42, and depletion of Cdc42 using siRNA abolishes TWEAK-induced Rac1 activation and abrogates glioma cell migration and invasion. In contrast, Rac1 depletion does not affect Cdc42 activation by Fn14, showing that Cdc42 mediates TWEAK-stimulated Rac1 activation. Furthermore, we identified two guanine nucleotide exchange factors (GEF), Ect2 and Trio, involved in TWEAK-induced activation of Cdc42 and Rac1, respectively. Depletion of Ect2 abrogates both TWEAK-induced Cdc42 and Rac1 activation, as well as subsequent TWEAK-Fn14–directed glioma cell migration and invasion. In contrast, Trio depletion inhibits TWEAK-induced Rac1 activation but not TWEAK-induced Cdc42 activation. Finally, inappropriate expression of Fn14 or Ect2 in mouse astrocytes in vivo using an RCAS vector system for glial-specific gene transfer in G-tva transgenic mice induces astrocyte migration within the brain, corroborating the in vitro importance of the TWEAK-Fn14 signaling cascade in glioblastoma invasion. Our results suggest that the TWEAK-Fn14 signaling axis stimulates glioma cell migration and invasion through two GEF-GTPase signaling units, Ect2-Cdc42 and Trio-Rac1. Components of the Fn14-Rho GEF-Rho GTPase signaling pathway present innovative drug targets for glioma therapy. PMID:22571869

Functional characterization and cellular dynamics of the CDC-42 - RAC - CDC-24 module in Neurospora crassa.

PubMed

Araujo-Palomares, Cynthia L; Richthammer, Corinna; Seiler, Stephan; Castro-Longoria, Ernestina

2011-01-01

Rho-type GTPases are key regulators that control eukaryotic cell polarity, but their role in fungal morphogenesis is only beginning to emerge. In this study, we investigate the role of the CDC-42 - RAC - CDC-24 module in Neurospora crassa. rac and cdc-42 deletion mutants are viable, but generate highly compact colonies with severe morphological defects. Double mutants carrying conditional and loss of function alleles of rac and cdc-42 are lethal, indicating that both GTPases share at least one common essential function. The defects of the GTPase mutants are phenocopied by deletion and conditional alleles of the guanine exchange factor (GEF) cdc-24, and in vitro GDP-GTP exchange assays identify CDC-24 as specific GEF for both CDC-42 and RAC. In vivo confocal microscopy shows that this module is organized as membrane-associated cap that covers the hyphal apex. However, the specific localization patterns of the three proteins are distinct, indicating different functions of RAC and CDC-42 within the hyphal tip. CDC-42 localized as confined apical membrane-associated crescent, while RAC labeled a membrane-associated ring excluding the region labeled by CDC42. The GEF CDC-24 occupied a strategic position, localizing as broad apical membrane-associated crescent and in the apical cytosol excluding the Spitzenkörper. RAC and CDC-42 also display distinct localization patterns during branch initiation and germ tube formation, with CDC-42 accumulating at the plasma membrane before RAC. Together with the distinct cellular defects of rac and cdc-42 mutants, these localizations suggest that CDC-42 is more important for polarity establishment, while the primary function of RAC may be maintaining polarity. In summary, this study identifies CDC-24 as essential regulator for RAC and CDC-42 that have common and distinct functions during polarity establishment and maintenance of cell polarity in N. crassa.

BIG1, a brefeldin A-inhibited guanine nucleotide-exchange protein regulates neurite development via PI3K-AKT and ERK signaling pathways.

PubMed

Zhou, C; Li, C; Li, D; Wang, Y; Shao, W; You, Y; Peng, J; Zhang, X; Lu, L; Shen, X

2013-12-19

The elongation of neuron is highly dependent on membrane trafficking. Brefeldin A (BFA)-inhibited guanine nucleotide-exchange protein 1 (BIG1) functions in the membrane trafficking between the Golgi apparatus and the plasma membrane. BFA, an uncompetitive inhibitor of BIG1 can inhibit neurite outgrowth and polarity development. In this study, we aimed to define the possible role of BIG1 in neurite development and to further investigate the potential mechanism. By immunostaining, we found that BIG1 was extensively colocalized with synaptophysin, a marker for synaptic vesicles in soma and partly in neurites. The amount of both protein and mRNA of BIG1 were up-regulated during rat brain development. BIG1 depletion significantly decreased the neurite length and inhibited the phosphorylation of phosphatidylinositide 3-kinase (PI3K) and protein kinase B (AKT). Inhibition of BIG1 guanine nucleotide-exchange factor (GEF) activity by BFA or overexpression of the dominant-negative BIG1 reduced PI3K and AKT phosphorylation, indicating regulatory effects of BIG1 on PI3K-AKT signaling pathway is dependent on its GEF activity. BIG1 siRNA or BFA treatment also significantly reduced extracellular signal-regulated kinase (ERK) phosphorylation. Overexpression of wild-type BIG1 significantly increased ERK phosphorylation, but the dominant-negative BIG1 had no effect on ERK phosphorylation, indicating the involvement of BIG1 in ERK signaling regulation may not be dependent on its GEF activity. Our result identified a novel function of BIG1 in neurite development. The newly recognized function integrates the function of BIG1 in membrane trafficking with the activation of PI3K-AKT and ERK signaling pathways which are critical in neurite development. Copyright © 2013 IBRO. Published by Elsevier Ltd. All rights reserved.

Why are SiX5(-) and GeX5(-) (X = F, Cl) stable but not CF5(-) and CCl5(-)?

PubMed

Marchaj, Marzena; Freza, Sylwia; Skurski, Piotr

2012-03-01

The possible existence of the CF(5)(-), CCl(5)(-), SiF(5)(-), SiCl(5)(-), GeF(5)(-), and GeCl(5)(-) anions has been investigated using ab initio methods. The species containing Si and Ge as central atoms were found to adopt the D(3h)-symmetry trigonal bipyramidal equilibrium structures whose thermodynamic stabilities were confirmed by examining the most probable fragmentation channels. The ab initio re-examination of the electronic stabilities of the SiF(5)(-), SiCl(5)(-), GeF(5)(-), and GeCl(5)(-) anions [using the OVGF(full) method with the 6-311+G(3df) basis set] led to the very large vertical electron detachment (VDE) energies of 9.316 eV (SiF(5)(-)) and 9.742 eV (GeF(5)(-)), whereas smaller VDEs of 6.196 and 6.452 eV were predicted for the SiCl(5)(-) and GeCl(5)(-) species, respectively. By contrast, the high-symmetry and structurally compact anionic CF(5)(-) and CCl(5)(-) systems cannot exist due to the strongly repulsive potential predicted for the X(-) (F(-) or Cl(-)) approaching the CX(4) (CF(4) or CCl(4)). The formation of weakly bound CX(4)···X(-) (CF(4)···F(-) and CCl(4)···Cl(-)) anionic complexes (consisting of pseudotetrahedral neutral CX(4) with the weakly tethered X(-)) might be expected at low temperatures (approaching 0 K), whereas neither CX(5)(-) (CF(5)(-), CCl(5)(-)) systems nor CX(4)···X(-) (CF(4)···F(-) and CCl(4)···Cl(-)) complexes can exist in the elevated temperatures (above 0K) due to their susceptibility to the fragmentation (leading to the X(-) loss). © 2012 American Chemical Society

Recurrent De Novo Mutations Disturbing the GTP/GDP Binding Pocket of RAB11B Cause Intellectual Disability and a Distinctive Brain Phenotype.

PubMed

Lamers, Ideke J C; Reijnders, Margot R F; Venselaar, Hanka; Kraus, Alison; Jansen, Sandra; de Vries, Bert B A; Houge, Gunnar; Gradek, Gyri Aasland; Seo, Jieun; Choi, Murim; Chae, Jong-Hee; van der Burgt, Ineke; Pfundt, Rolph; Letteboer, Stef J F; van Beersum, Sylvia E C; Dusseljee, Simone; Brunner, Han G; Doherty, Dan; Kleefstra, Tjitske; Roepman, Ronald

2017-11-02

The Rab GTPase family comprises ∼70 GTP-binding proteins, functioning in vesicle formation, transport and fusion. They are activated by a conformational change induced by GTP-binding, allowing interactions with downstream effectors. Here, we report five individuals with two recurrent de novo missense mutations in RAB11B; c.64G>A; p.Val22Met in three individuals and c.202G>A; p.Ala68Thr in two individuals. An overlapping neurodevelopmental phenotype, including severe intellectual disability with absent speech, epilepsy, and hypotonia was observed in all affected individuals. Additionally, visual problems, musculoskeletal abnormalities, and microcephaly were present in the majority of cases. Re-evaluation of brain MRI images of four individuals showed a shared distinct brain phenotype, consisting of abnormal white matter (severely decreased volume and abnormal signal), thin corpus callosum, cerebellar vermis hypoplasia, optic nerve hypoplasia and mild ventriculomegaly. To compare the effects of both variants with known inactive GDP- and active GTP-bound RAB11B mutants, we modeled the variants on the three-dimensional protein structure and performed subcellular localization studies. We predicted that both variants alter the GTP/GDP binding pocket and show that they both have localization patterns similar to inactive RAB11B. Evaluation of their influence on the affinity of RAB11B to a series of binary interactors, both effectors and guanine nucleotide exchange factors (GEFs), showed induction of RAB11B binding to the GEF SH3BP5, again similar to inactive RAB11B. In conclusion, we report two recurrent dominant mutations in RAB11B leading to a neurodevelopmental syndrome, likely caused by altered GDP/GTP binding that inactivate the protein and induce GEF binding and protein mislocalization. Copyright © 2017 American Society of Human Genetics. All rights reserved.

Practical implementation of a particle filter data assimilation approach to estimate initial hydrologic conditions and initialize medium-range streamflow forecasts

NASA Astrophysics Data System (ADS)

Clark, E.; Wood, A.; Nijssen, B.; Newman, A. J.; Mendoza, P. A.

2016-12-01

The System for Hydrometeorological Applications, Research and Prediction (SHARP), developed at the National Center for Atmospheric Research (NCAR), University of Washington, U.S. Army Corps of Engineers, and U.S. Bureau of Reclamation, is a fully automated ensemble prediction system for short-term to seasonal applications. It incorporates uncertainty in initial hydrologic conditions (IHCs) and in hydrometeorological predictions. In this implementation, IHC uncertainty is estimated by propagating an ensemble of 100 plausible temperature and precipitation time series through the Sacramento/Snow-17 model. The forcing ensemble explicitly accounts for measurement and interpolation uncertainties in the development of gridded meteorological forcing time series. The resulting ensemble of derived IHCs exhibits a broad range of possible soil moisture and snow water equivalent (SWE) states. To select the IHCs that are most consistent with the observations, we employ a particle filter (PF) that weights IHC ensemble members based on observations of streamflow and SWE. These particles are then used to initialize ensemble precipitation and temperature forecasts downscaled from the Global Ensemble Forecast System (GEFS), generating a streamflow forecast ensemble. We test this method in two basins in the Pacific Northwest that are important for water resources management: 1) the Green River upstream of Howard Hanson Dam, and 2) the South Fork Flathead River upstream of Hungry Horse Dam. The first of these is characterized by mixed snow and rain, while the second is snow-dominated. The PF-based forecasts are compared to forecasts based on a single IHC (corresponding to median streamflow) paired with the full GEFS ensemble, and 2) the full IHC ensemble, without filtering, paired with the full GEFS ensemble. In addition to assessing improvements in the spread of IHCs, we perform a hindcast experiment to evaluate the utility of PF-based data assimilation on streamflow forecasts at 1- to 7-day lead times.

Steroid markers to assess sewage and other sources of organic contaminants in surface sediments of Cienfuegos Bay, Cuba.

PubMed

Tolosa, I; Mesa, M; Alonso-Hernandez, C M

2014-09-15

Analyses of faecal steroids in coastal sediments from Cienfuegos Bay Cuba indicate chronic sewage contamination at the main outfalls from the city, where concentrations of coprostanol up to 5400ngg(-)(1) (dry wt) were measured. In contrast, steroid concentrations and compositions from sites from the south part of the Bay are characteristic of uncontaminated sewage environments. The levels of coprostanol in the Cienfuegos sediments compares to the lower to mid-range of concentrations reported for coastal sediments on a world-wide basis, with sedimentary levels markedly below those previously reported for heavily impacted sites. This study delivers baseline data for further investigation of the effectiveness of the proposed sewerage plan promoted by the GEF project in Cienfuegos. Investigations on the correlations between faecal steroids and other organic contaminants confirmed that the major source of petroleum hydrocarbons within the bay was associated with the sewage effluents from the Cienfuegos city. Copyright © 2014 Elsevier Ltd. All rights reserved.

Global scale predictability of floods

NASA Astrophysics Data System (ADS)

Weerts, Albrecht; Gijsbers, Peter; Sperna Weiland, Frederiek

2016-04-01

Flood (and storm surge) forecasting at the continental and global scale has only become possible in recent years (Emmerton et al., 2016; Verlaan et al., 2015) due to the availability of meteorological forecast, global scale precipitation products and global scale hydrologic and hydrodynamic models. Deltares has setup GLOFFIS a research-oriented multi model operational flood forecasting system based on Delft-FEWS in an open experimental ICT facility called Id-Lab. In GLOFFIS both the W3RA and PCRGLOB-WB model are run in ensemble mode using GEFS and ECMWF-EPS (latency 2 days). GLOFFIS will be used for experiments into predictability of floods (and droughts) and their dependency on initial state estimation, meteorological forcing and the hydrologic model used. Here we present initial results of verification of the ensemble flood forecasts derived with the GLOFFIS system. Emmerton, R., Stephens, L., Pappenberger, F., Pagano, T., Weerts, A., Wood, A. Salamon, P., Brown, J., Hjerdt, N., Donnelly, C., Cloke, H. Continental and Global Scale Flood Forecasting Systems, WIREs Water (accepted), 2016 Verlaan M, De Kleermaeker S, Buckman L. GLOSSIS: Global storm surge forecasting and information system 2015, Australasian Coasts & Ports Conference, 15-18 September 2015,Auckland, New Zealand.

Production, Service and Trade Enterprise EKOREX Co. Ltd.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wlodkowski, A.

1995-12-31

In the first period of its activity the business employed skilled and experienced specialists from the ex-Military College for Army Chemical Engineers in Cracow; therefore, the enterprise dealt chiefly with the elimination of environmental contamination. Nowadays, the enterprise`s operational range comprises: consulting and training services related with ecology; study on environmental contamination; participation in the US program of low emission elimination in Cracow; designing, consulting in the realization of projects {open_quotes}GEF{close_quotes} (Global Environmental Facility); designing, construction, servicing, operating the sewerage and water treatment plants, boiler-houses, incinerators etc.; and designing of heat networks, exchanger junctions, central heating and household hot watermore » installations. Since 1991 employees have individually participated in making the program and in testing boilers and fuels verified in the boiler houses covered by the Polish - US program of reduction of low emission sources in Cracow. We have actively joined the program of elimination of heating network boiler houses (industrial and local) by designing (for the Cracow cogeneration plant and MPEC) new connections among some structures and the municiple thermal distribution network and exchangers stations. In 1994, 47 such designs were made and have been working on successive projects to be carried out in Cracow.« less

Regulation of ATM-Dependent DNA Damage Responses in Breast Cancer by the RhoGEF Net1

DTIC Science & Technology

2015-05-01

mediators of gastric cancer. Br. J. Cancer 94(8):1204-1212. 11. Shen SQ, et al. 2008 Expression and clinical significance of NET-1 and PCNA in...ATM mutations and phenotypes in Ataxia-telangiectasia families in the british isles: Expression of mutant ATM and the risk of Leukemia, Lymphoma

Regulation of ATM-Dependent DNA Damage Responses in Breast Cancer by the RhoGEF Net1

DTIC Science & Technology

2014-04-01

1998) Science 279: 509-514. 12. Harper JW, et al., (2007) The DNA Damage Response: Ten years after. Mol. Cell 28; 739-745. 13. Hill R, et al., (2010...RhoGTPases: Biochemistry and Biology. Annu. Rev. Cell Dev. Biol. 21:247-269. 17. Khanna KK, et al., (2001) ATM, a central controller of cellular

Building District Capacity for System-Wide Instructional Improvement in Stamford Public Schools. Working Paper. GE Foundation "Developing Futures"™ in Education Evaluation Series

ERIC Educational Resources Information Center

Fink, Ryan; Riggan, Matt

2013-01-01

This report summarizes findings from one component of the Consortium for Policy Research in Education's (CPRE) evaluation of the General Electric Foundation's (GEF) "Developing Futures"™ in Education program in Stamford Public Schools (SPS). The purpose was to closely analyze district capacity to support system-wide instructional…

Building District Capacity for System-Wide Instructional Improvement in Cincinnati Public Schools. Working Paper. GE Foundation "Developing Futures"™ in Education Evaluation Series

ERIC Educational Resources Information Center

Sam, Cecile; Riggan, Matt

2013-01-01

This report summarizes findings from one component of the Consortium for Policy Research in Education's (CPRE) evaluation of the General Electric Foundation's (GEF) "Developing Futures"™ in Education program in Cincinnati Public Schools (CPS). The purpose was to closely analyze district capacity to support system-wide instructional…

Activation of multiple signaling pathways causes developmental defects in mice with a Noonan syndrome–associated Sos1 mutation

PubMed Central

Chen, Peng-Chieh; Wakimoto, Hiroko; Conner, David; Araki, Toshiyuki; Yuan, Tao; Roberts, Amy; Seidman, Christine E.; Bronson, Roderick; Neel, Benjamin G.; Seidman, Jonathan G.; Kucherlapati, Raju

2010-01-01

Noonan syndrome (NS) is an autosomal dominant genetic disorder characterized by short stature, unique facial features, and congenital heart disease. About 10%–15% of individuals with NS have mutations in son of sevenless 1 (SOS1), which encodes a RAS and RAC guanine nucleotide exchange factor (GEF). To understand the role of SOS1 in the pathogenesis of NS, we generated mice with the NS-associated Sos1E846K gain-of-function mutation. Both heterozygous and homozygous mutant mice showed many NS-associated phenotypes, including growth delay, distinctive facial dysmorphia, hematologic abnormalities, and cardiac defects. We found that the Ras/MAPK pathway as well as Rac and Stat3 were activated in the mutant hearts. These data provide in vivo molecular and cellular evidence that Sos1 is a GEF for Rac under physiological conditions and suggest that Rac and Stat3 activation might contribute to NS phenotypes. Furthermore, prenatal administration of a MEK inhibitor ameliorated the embryonic lethality, cardiac defects, and NS features of the homozygous mutant mice, demonstrating that this signaling pathway might represent a promising therapeutic target for NS. PMID:21041952

Thrombin Promotes Sustained Signaling and Inflammatory Gene Expression through the CDC25 and Ras-associating Domains of Phospholipase Cϵ*

PubMed Central

Dusaban, Stephanie S.; Kunkel, Maya T.; Smrcka, Alan V.; Brown, Joan Heller

2015-01-01

Phospholipase C-epsilon (PLCϵ) plays a critical role in G-protein-coupled receptor-mediated inflammation. In addition to its ability to generate the second messengers inositol 1,4,5-trisphosphate and diacylglycerol, PLCϵ, unlike the other phospholipase C family members, is activated in a sustained manner. We hypothesized that the ability of PLCϵ to function as a guanine nucleotide exchange factor (GEF) for Rap1 supports sustained downstream signaling via feedback of Rap1 to the enzyme Ras-associating (RA2) domain. Using gene deletion and adenoviral rescue, we demonstrate that both the GEF (CDC25 homology domain) and RA2 domains of PLCϵ are required for long term protein kinase D (PKD) activation and subsequent induction of inflammatory genes. PLCϵ localization is largely intracellular and its compartmentalization could contribute to its sustained activation. Here we show that localization of PLCϵ to the Golgi is required for activation of PKD in this compartment as well as for subsequent induction of inflammatory genes. These data provide a molecular mechanism by which PLCϵ mediates sustained signaling and by which astrocytes mediate pathophysiological inflammatory responses. PMID:26350460

Multiplex image-based autophagy RNAi screening identifies SMCR8 as ULK1 kinase activity and gene expression regulator

PubMed Central

Jung, Jennifer; Nayak, Arnab; Schaeffer, Véronique; Starzetz, Tatjana; Kirsch, Achim K; Müller, Stefan; Dikic, Ivan; Mittelbronn, Michel; Behrends, Christian

2017-01-01

Autophagy is an intracellular recycling and degradation pathway that depends on membrane trafficking. Rab GTPases are central for autophagy but their regulation especially through the activity of Rab GEFs remains largely elusive. We employed a RNAi screen simultaneously monitoring different populations of autophagosomes and identified 34 out of 186 Rab GTPase, GAP and GEF family members as potential autophagy regulators, amongst them SMCR8. SMCR8 uses overlapping binding regions to associate with C9ORF72 or with a C9ORF72-ULK1 kinase complex holo-assembly, which function in maturation and formation of autophagosomes, respectively. While focusing on the role of SMCR8 during autophagy initiation, we found that kinase activity and gene expression of ULK1 are increased upon SMCR8 depletion. The latter phenotype involved association of SMCR8 with the ULK1 gene locus. Global mRNA expression analysis revealed that SMCR8 regulates transcription of several other autophagy genes including WIPI2. Collectively, we established SMCR8 as multifaceted negative autophagy regulator. DOI: http://dx.doi.org/10.7554/eLife.23063.001 PMID:28195531

The Molecular Motor KIF21B Mediates Synaptic Plasticity and Fear Extinction by Terminating Rac1 Activation.

PubMed

Morikawa, Momo; Tanaka, Yosuke; Cho, Hyun-Soo; Yoshihara, Masaharu; Hirokawa, Nobutaka

2018-06-26

Fear extinction is a component of cognitive flexibility that is relevant for important psychiatric diseases, but its molecular mechanism is still largely elusive. We established mice lacking the kinesin-4 motor KIF21B as a model for fear extinction defects. Postsynaptic NMDAR-dependent long-term depression (LTD) is specifically impaired in knockouts. NMDAR-mediated LTD-causing stimuli induce dynamic association of KIF21B with the Rac1GEF subunit engulfment and cell motility protein 1 (ELMO1), leading to ELMO1 translocation out of dendritic spines and its sequestration in endosomes. This process may essentially terminate transient activation of Rac1, shrink spines, facilitate AMPAR endocytosis, and reduce postsynaptic strength, thereby forming a mechanistic link to LTD expression. Antagonizing ELMO1/Dock Rac1GEF activity by the administration of 4-[3'-(2″-chlorophenyl)-2'-propen-1'-ylidene]-1-phenyl-3,5-pyrazolidinedione (CPYPP) significantly reverses the knockout phenotype. Therefore, we propose that KIF21B-mediated Rac1 inactivation is a key molecular event in NMDAR-dependent LTD expression underlying cognitive flexibility in fear extinction. Copyright © 2018 The Author(s). Published by Elsevier Inc. All rights reserved.

Rab14 and Its Exchange Factor FAM116 Link Endocytic Recycling and Adherens Junction Stability in Migrating Cells

PubMed Central

Linford, Andrea; Yoshimura, Shin-ichiro; Bastos, Ricardo Nunes; Langemeyer, Lars; Gerondopoulos, Andreas; Rigden, Daniel J.; Barr, Francis A.

2012-01-01

Summary Rab GTPases define the vesicle trafficking pathways underpinning cell polarization and migration. Here, we find that Rab4, Rab11, and Rab14 and the candidate Rab GDP-GTP exchange factors (GEFs) FAM116A and AVL9 are required for cell migration. Rab14 and its GEF FAM116A localize to and act on an intermediate compartment of the transferrin-recycling pathway prior to Rab11 and after Rab5 and Rab4. This Rab14 intermediate recycling compartment has specific functions in migrating cells discrete from early and recycling endosomes. Rab14-depleted cells show increased N-cadherin levels at junctional complexes and cannot resolve cell-cell junctions. This is due to decreased shedding of cell-surface N-cadherin by the ADAM family protease ADAM10/Kuzbanian. In FAM116A- and Rab14-depleted cells, ADAM10 accumulates in a transferrin-positive endocytic compartment, and the cell-surface level of ADAM10 is correspondingly reduced. FAM116 and Rab14 therefore define an endocytic recycling pathway needed for ADAM protease trafficking and regulation of cell-cell junctions. PMID:22595670

[Effects of recruitment maneuver in prone position on hemodynamics in patients with severe pulmonary infection].

PubMed

Fan, Yuan-hua; Liu, Yuan-fei; Zhu, Hua-yong; Zhang, Min

2012-02-01

To evaluate effects of recruitment maneuver in prone position on hemodynamics in patients with severe pulmonary infection, based on the protective pulmonary ventilation strategy. Ninety-seven cases with severe pulmonary infection admitted to intensive care unit (ICU) of Ganzhou City People's Hospital undergoing mechanical ventilation were involved. Volume controlled ventilation mode with small tidal volume (8 ml/kg) and positive end-expiratory pressure (PEEP) of 6 cm H(2)O [1 cm H(2)O = 0.098 kPa] was conducted. Each patient underwent recruitment maneuver in supine position and then in prone position [PEEP 20 cm H(2)O+pressure control (PC) 20 cm H(2)O]. Heart rate (HR), mean arterial pressure (MAP), pulse oxygen saturation [SpO(2)] and blood gas analysis data were recorded before and after recruitment maneuver in either position. A double-lumen venous catheter was inserted into internal jugular vein or subclavian vein, and a pulse index contour cardiac output (PiCCO) catheter was introduced into femoral artery. Cardiac index (CI), stroke volume index (SVI), systemic vascular resistance index (SVRI), intra-thoracic blood volume index (ITBVI), extra vascular lung water index (EVLWI), global end-diastolic volume index (GEDVI), global ejection fraction (GEF), stroke volume variation (SVV) and central vein pressure (CVP) were monitored. (1) Compared with data before recruitment maneuver, there were no significant differences in HR and MAP after supine position and prone position recruitment maneuver, but significant differences in SpO(2) were found between before and after recruitment maneuver when patients' position was changed (supine position: 0.954 ± 0.032 vs. 0.917 ± 0.025, P < 0.05; prone position: 0.982 ± 0.028 vs. 0.936 ± 0.039, P < 0.05). SpO(2) was higher in prone position recruitment maneuver (P < 0.05). (2) Compared with data before recruitment maneuver, CI [L×min(-1)×m(-2)], SVI (ml/m(2)), GEDVI (ml/m(2)) and GEF were decreased significantly during recruitment maneuver (supine position: CI 3.2 ± 0.4 vs. 3.8 ± 0.6, SVI 32.4 ± 5.6 vs. 38.8 ± 6.5, GEDVI 689 ± 44 vs. 766 ± 32, GEF 0.267 ± 0.039 vs. 0.305 ± 0.056; prone position: CI 3.1 ± 0.5 vs. 3.6 ± 0.4, SVI 31.2 ± 5.8 vs. 37.3 ± 5.0, GEDVI 678 ± 41 vs. 758 ± 36, GEF 0.268 ± 0.040 vs. 0.288 ± 0.053, all P < 0.05), and CVP [cm H(2)O] and SVV were significantly increased [supine position: CVP 10.7 ± 1.5 vs. 8.2 ± 2.5, SVV (11.2 ± 3.3)% vs. (8.3 ± 4.7)%; prone position: CVP 10.3 ± 1.8 vs. 8.1 ± 2.5, SVV (12.7 ± 3.4)% vs. (9.1 ± 3.6)%, all P < 0.05], but they returned to the level of that before recruitment maneuver soon after termination of recruitment maneuver. There were no significant differences in SVRI, ITBVI and EVLWI between before and after recruitment maneuver in both positions. There were also no significant differences in above parameters between two positions. Based on the lung protective ventilation strategy of small tidal volume with PEEP, oxygenation was improved and SpO(2) was increased significantly when prone position ventilation combined with lung recruitment method was used in severe pulmonary infection patients. The effect of recruitment maneuver during prone position on hemodynamics was slight, except a temporary decrease of SVI and GEF just during recruitment maneuver.

Building District Capacity for System-Wide Instructional Improvement in Erie Public Schools. Working Paper. GE Foundation "Developing Futures"™ in Education Evaluation Series

ERIC Educational Resources Information Center

Riggan, Matt; Fink, Ryan; Sam, Cecile; Darfler, Anne

2013-01-01

This report summarizes findings from one component of the Consortium for Policy Research in Education's (CPRE) evaluation of the General Electric Foundation's (GEF) "Developing Futures" ™ in Education program in Erie Public Schools (EPS). As described in the CPRE proposal and research design, the purpose was to closely analyze district…

Building District Capacity for System-Wide Instructional Improvement in Jefferson County Public Schools. Working Paper. GE Foundation "Developing Futures"™ in Education Evaluation Series

ERIC Educational Resources Information Center

Darfler, Anne; Riggan, Matt

2013-01-01

This report summarizes findings from one component of the Consortium for Policy Research in Education's (CPRE) evaluation of the General Electric Foundation's (GEF) "Developing Futures"™ in Education program in Jefferson County Public Schools (JCPS). As described in the CPRE proposal and research design, the purpose was to closely…

Viral Activation of MK2-hsp27-p115RhoGEF-RhoA Signaling Axis Causes Cytoskeletal Rearrangements, P-body Disruption and ARE-mRNA Stabilization

PubMed Central

Corcoran, Jennifer A.; Johnston, Benjamin P.; McCormick, Craig

2015-01-01

Kaposi's sarcoma-associated herpesvirus (KSHV) is the infectious cause of several AIDS-related cancers, including the endothelial cell (EC) neoplasm Kaposi's sarcoma (KS). KSHV-infected ECs secrete abundant host-derived pro-inflammatory molecules and angiogenic factors that contribute to tumorigenesis. The precise contributions of viral gene products to this secretory phenotype remain to be elucidated, but there is emerging evidence for post-transcriptional regulation. The Kaposin B (KapB) protein is thought to contribute to the secretory phenotype in infected cells by binding and activating the stress-responsive kinase MK2, thereby selectively blocking decay of AU-rich mRNAs (ARE-mRNAs) encoding pro-inflammatory cytokines and angiogenic factors. Processing bodies (PBs) are cytoplasmic ribonucleoprotein foci in which ARE-mRNAs normally undergo rapid 5′ to 3′ decay. Here, we demonstrate that PB dispersion is a feature of latent KSHV infection, which is dependent on kaposin protein expression. KapB is sufficient to disperse PBs, and KapB-mediated ARE-mRNA stabilization could be partially reversed by treatments that restore PBs. Using a combination of genetic and chemical approaches we provide evidence that KapB-mediated PB dispersion is dependent on activation of a non-canonical Rho-GTPase signaling axis involving MK2, hsp27, p115RhoGEF and RhoA. PB dispersion in latently infected cells is likewise dependent on p115RhoGEF. In addition to PB dispersion, KapB-mediated RhoA activation in primary ECs caused actin stress fiber formation, increased cell motility and angiogenesis; these effects were dependent on the activity of the RhoA substrate kinases ROCK1/2. By contrast, KapB-mediated PB dispersion occurred in a ROCK1/2-independent manner. Taken together, these observations position KapB as a key contributor to viral reprogramming of ECs, capable of eliciting many of the phenotypes characteristic of KS tumor cells, and strongly contributing to the post-transcriptional control of EC gene expression and secretion. PMID:25569678

Cross-Species Analyses Identify the BNIP-2 and Cdc42GAP Homology (BCH) Domain as a Distinct Functional Subclass of the CRAL_TRIO/Sec14 Superfamily

PubMed Central

Gupta, Anjali Bansal; Wee, Liang En; Zhou, Yi Ting; Hortsch, Michael; Low, Boon Chuan

2012-01-01

The CRAL_TRIO protein domain, which is unique to the Sec14 protein superfamily, binds to a diverse set of small lipophilic ligands. Similar domains are found in a range of different proteins including neurofibromatosis type-1, a Ras GTPase-activating Protein (RasGAP) and Rho guanine nucleotide exchange factors (RhoGEFs). Proteins containing this structural protein domain exhibit a low sequence similarity and ligand specificity while maintaining an overall characteristic three-dimensional structure. We have previously demonstrated that the BNIP-2 and Cdc42GAP Homology (BCH) protein domain, which shares a low sequence homology with the CRAL_TRIO domain, can serve as a regulatory scaffold that binds to Rho, RhoGEFs and RhoGAPs to control various cell signalling processes. In this work, we investigate 175 BCH domain-containing proteins from a wide range of different organisms. A phylogenetic analysis with ∼100 CRAL_TRIO and similar domains from eight representative species indicates a clear distinction of BCH-containing proteins as a novel subclass within the CRAL_TRIO/Sec14 superfamily. BCH-containing proteins contain a hallmark sequence motif R(R/K)h(R/K)(R/K)NL(R/K)xhhhhHPs (‘h’ is large and hydrophobic residue and ‘s’ is small and weekly polar residue) and can be further subdivided into three unique subtypes associated with BNIP-2-N, macro- and RhoGAP-type protein domains. A previously unknown group of genes encoding ‘BCH-only’ domains is also identified in plants and arthropod species. Based on an analysis of their gene-structure and their protein domain context we hypothesize that BCH domain-containing genes evolved through gene duplication, intron insertions and domain swapping events. Furthermore, we explore the point of divergence between BCH and CRAL-TRIO proteins in relation to their ability to bind small GTPases, GAPs and GEFs and lipid ligands. Our study suggests a need for a more extensive analysis of previously uncharacterized BCH, ‘BCH-like’ and CRAL_TRIO-containing proteins and their significance in regulating signaling events involving small GTPases. PMID:22479462

Up-regulation of the RhoA/Rho-kinase signaling pathway in corpus cavernosum from endothelial nitric-oxide synthase (NOS), but not neuronal NOS, null mice.

PubMed

Priviero, Fernanda B M; Jin, Li-Ming; Ying, Zhekang; Teixeira, Cleber E; Webb, R Clinton

2010-04-01

We tested the hypothesis that the basal release of nitric oxide (NO) from endothelial cells modulates contractile activity in the corpus cavernosum (CC) via inhibition of the RhoA/Rho-kinase signaling pathway. Cavernosal strips from wild-type (WT), endothelial nitric-oxide synthase knockout [eNOS(-/-)], and neuronal nitric-oxide synthase knockout [nNOS(-/-)] mice were mounted in myographs, and isometric force was recorded. mRNA and protein expression of key molecules in the RhoA/Rho-kinase pathway were analyzed by real-time polymerase chain reaction and Western blot, respectively. The cGMP levels were determined. The Rho-kinase inhibitors (R)-(+)-trans-N-(4-pyridyl)-4-(1-aminoethyl)-cyclohexanecarboxamide (Y-27632) and (S)-(+)-2-methyl-1-[(4-methyl-5-isoquinolinyl)sulfonyl] homopiperazine (H-1152) reduced cavernosal contractions evoked by phenylephrine or electrical field stimulation (EFS) in a concentration-dependent manner, although this inhibition was less effective in tissues from eNOS(-/-) mice. Y-27632 enhanced relaxations induced by sodium nitroprusside, EFS, and NO (administered as acidified NaNO2) without affecting the cGMP content of the cavernosal strips. This enhancement was less prominent in CC from eNOS(-/-). The protein expression of RhoA, Rho-guanine dissociation inhibitor, and Rho-kinase beta did not differ among the strains. However, in eNOS(-/-) CC, the protein expression of Rho-kinase alpha and both mRNA and protein expression of p115-Rho-associated guanine exchange factor (RhoGEF), PDZ-RhoGEF, and leukemia-associated RhoGEF were up-regulated. Phosphorylation of MYPT1 at Thr696 was higher in tissues from eNOS(-/-) mice. A high concentration of Y-27632 significantly enhanced NO release in CC stimulated by EFS. These results suggest a basal release of NO from endothelial cells, which inhibits contractions mediated by the RhoA/Rho-kinase pathway and modulates the expression of proteins related to this pathway in mouse CC. It indicates that endothelial integrity is essential to the maintenance of erectile function.

[RS-1 enhanced the efficiency of CRISPR-Cas9 mediated knock-in of human lactoferrin].

PubMed

Zhou, Wenjun; Guo, Rihong; Deng, Mingtian; Wang, Feng; Zhang, Yanli

2017-08-25

This study aims to knock out the goat β-lactoglobulin (BLG) gene using CRISPR-Cas9 system and knock in human lactoferrin (hLF) at the BLG locus, and further study the effect of RAD51 stimulatory compound (RS-1) on homologous recombination efficiency. First, we designed an sgRNA targeting the first exon of goat BLG gene and constructed a co-expression vector pCas9-sgBLG. This sgRNA vector was then transfected into goat ear fibroblasts (GEFs), and the target region was examined by T7EN1 assay and sequencing. Second, we constructed a targeting vector pBHA-hLF-NIE including NEO and EGFP genes based on BLG gene locus. This targeting vector together with pCas9-sgBLG expression vector was co-transfected into GEFs. Transfected cells were then treated with 0, 5, 10 and 20 μmol/L RS-1 for 72 h to analyse the EGFP expression efficiency. Next, we used 800 μg/mL G418 to screen G418-resistent cell clones, and studied hLF site-specific knock-in cell clones by PCR and sequencing. The editing efficiency of sgBLG was between 25% and 31%. The EGFP expression efficiency indicated that the gene knock-in efficiency was improved by RS-1 in a dose-dependent manner, which could reach 3.5-fold compared to the control group. The percentage of positive cells with hLF knock-in was increased to 32.61% when 10 μmol/L RS-1 was used. However, when the concentration of RS-1 increased to 20 μmol/L, the percentage of positive cells decreased to 22.22% and resulted in an increase of senescent cell clone number. These results suggested that hLF knock-in and BLG knock-out in GEFs were achieved by using CRISPR/Cas9 system, and optimum concentration of RS-1 could improve knock-in efficiency, which provides a reference for efficiently obtaining gene knock-in cells using CRISPR/Cas9 in the future.

Genetic evidence of 'genuine' empty follicle syndrome: a novel effective mutation in the LHCGR gene and review of the literature.

PubMed

Yuan, Ping; He, Zuyong; Zheng, Lingyan; Wang, Wenjun; Li, Yu; Zhao, Haijing; Zhang, Victor Wei; Zhang, Qingxue; Yang, Dongzi

2017-04-01

Empty follicle syndrome (EFS) is a reproductive disorder in which no oocytes are retrieved during IVF. The existence of genuine EFS (GEFS) is still controversial, and to date, only one missense mutation of Luteinizing Hormone/Choriogonadotropin Receptor (LHCGR) has been reported to be associated with this disease. Here, we describe a GEFS patient in a non-consanguineous family from China. A 27-year-old woman presented with a 5-year history of primary infertility and LH resistance-like ovaries of unequal sizes, but with normal levels of circulating LH. In spite of a satisfactory ovarian reserve and response, no oocytes were retrieved after two cycles of IVF. Her condition did not appear to be failure of the hCG injection. It is more likely to be a genetic cause. A novel homozygous mutation in LHCGR gene, c.1345G>A (p.Ala449Thr), was detected in this patient. Each of her parents is heterozygous for this change, and the change was absent from 407 control subjects. Alanine at this amino acid position was highly conserved and replacement of threonine was predicted to disrupt the third transmembrane helix of the rhodopsin-like G protein-coupled receptor domain. Protein localization studies revealed that a portion of the mutant LHCGR protein molecules was retained intracellularly. Signalling studies demonstrated that this mutation had differing effects on the response of LHCGR to hCG or LH at different concentrations. Specifically, at a concentration <1 IU/ml, the mutant was activated by hCG stimulation but partially resistant to LH stimulation; at a higher concentration (>1 IU/ml), the mutant was activated by both hCG and LH. These data suggest that screening for mutations in the LHCGR gene may assist in the diagnosis of patients with GEFS. The literature describing the relationship between phenotype and genotypes in females is reviewed, and possible aetiologies and treatment options for this disease are proposed based on our and other studies. © The Author 2017. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Measurement of the 234U(n, f ) cross-section with quasi-monoenergetic beams in the keV and MeV range using a Micromegas detector assembly

NASA Astrophysics Data System (ADS)

Stamatopoulos, A.; Kanellakopoulos, A.; Kalamara, A.; Diakaki, M.; Tsinganis, A.; Kokkoris, M.; Michalopoulou, V.; Axiotis, M.; Lagoyiannis, A.; Vlastou, R.

2018-01-01

The 234U neutron-induced fission cross-section has been measured at incident neutron energies of 452, 550, 651 keV and 7.5, 8.7, 10 MeV using the 7Li ( p, n) and the 2H( d, n) reactions, respectively, relative to the 235U( n, f ) and 238U( n, f ) reference reactions. The measurement was performed at the neutron beam facility of the National Center for Scientific Research "Demokritos", using a set-up based on Micromegas detectors. The active mass of the actinide samples and the corresponding impurities were determined via α-spectroscopy using a surface barrier silicon detector. The neutron spectra intercepted by the actinide samples have been thoroughly studied by coupling the NeuSDesc and MCNP5 codes, taking into account the energy and angular straggling of the primary ion beams in the neutron source targets in addition to contributions from competing reactions ( e.g. deuteron break-up) and neutron scattering in the surrounding materials. Auxiliary Monte Carlo simulations were performed making combined use of the FLUKA and GEF codes, focusing particularly on the determination of the fission fragment detection efficiency. The developed methodology and the final results are presented.

Tight junctions negatively regulate mechanical forces applied to adherens junctions in vertebrate epithelial tissue.

PubMed

Hatte, Guillaume; Prigent, Claude; Tassan, Jean-Pierre

2018-02-05

Epithelia are layers of polarised cells tightly bound to each other by adhesive contacts. Epithelia act as barriers between an organism and its external environment. Understanding how epithelia maintain their essential integrity while remaining sufficiently plastic to allow events such as cytokinesis to take place is a key biological problem. In vertebrates, the remodelling and reinforcement of adherens junctions maintains epithelial integrity during cytokinesis. The involvement of tight junctions in cell division, however, has remained unexplored. Here, we examine the role of tight junctions during cytokinesis in the epithelium of the Xenopus laevis embryo. Depletion of the tight junction-associated proteins ZO-1 and GEF-H1 leads to altered cytokinesis duration and contractile ring geometry. Using a tension biosensor, we show that cytokinesis defects originate from misregulation of tensile forces applied to adherens junctions. Our results reveal that tight junctions regulate mechanical tension applied to adherens junctions, which in turn impacts cytokinesis.This article has an associated First Person interview with the first author of the paper. © 2018. Published by The Company of Biologists Ltd.

Role of the U.S. Military in the Professionalization of the Armed Forces of Liberia

DTIC Science & Technology

2015-06-12

professionalizing the AFL. The GEF states, “Partner nations provide for their own security, contribute effectively to broader regional or global security...facilitates the development of important professional and personal relationships that effectively strengthen military alliances and the international...Forces of Liberia (AFL) presents a developing opportunity to contribute to the broader U.S. interests. However, in order to ensure stability within

Vav1: Friend and Foe of Cancer.

PubMed

Guo, Fukun; Zheng, Yi

2017-12-01

A recent study shows that the protumorigenic guanine nucleotide exchange factor (GEF) Vav1 functions as a tumor suppressor in T cell acute lymphoblastic leukemia (T-ALL) through its ability to complex with the Cbl-b ubiquitin ligase and the intracellular domain of Notch1 (ICN1) and to promote ICN1 degradation. Vav1can act as a double-edged sword in tumorigenesis. Copyright © 2017 Elsevier Ltd. All rights reserved.

Rapid kinetic BRET measurements to monitor G protein activation by GPCR and non-GPCR proteins.

PubMed

Maziarz, Marcin; Garcia-Marcos, Mikel

2017-01-01

Heterotrimeric G proteins are central hubs of signal transduction whose activity is controlled by G protein-coupled receptors (GPCRs) as well as by a complex network of regulatory proteins. Recently, bioluminescence resonance energy transfer (BRET)-based assays have been used to monitor real-time activation of heterotrimeric G proteins in cells. Here we describe the use of a previously established BRET assay to monitor G protein activation upon GPCR stimulation and its adaptation to measure G protein activation by non-GPCR proteins, such as by cytoplasmic guanine nucleotide exchange factors (GEFs) like GIV/Girdin. The BRET assay monitors the release of free Gβγ from Gα-Gβγ heterotrimers as a readout of G protein activation, which is readily observable upon agonist stimulation of GPCRs. To control the signal input for non-GPCR activators, we describe the use of a chemically induced dimerization strategy to promote rapid membrane translocation of proteins containing the Gα-binding and -activating (GBA) motif found in some nonreceptor GEFs. The assay described here allows the kinetic measurement of G protein activation with subsecond temporal resolution and to compare the levels of activation induced by GPCR agonists vs those induced by the membrane recruitment of nonreceptor G protein signaling activators. © 2017 Elsevier Inc. All rights reserved.

Analysis of interphase node proteins in fission yeast by quantitative and superresolution fluorescence microscopy

PubMed Central

Akamatsu, Matthew; Lin, Yu; Bewersdorf, Joerg; Pollard, Thomas D.

2017-01-01

We used quantitative confocal microscopy and FPALM superresolution microscopy of live fission yeast to investigate the structures and assembly of two types of interphase nodes—multiprotein complexes associated with the plasma membrane that merge together and mature into the precursors of the cytokinetic contractile ring. During the long G2 phase of the cell cycle, seven different interphase node proteins maintain constant concentrations as they accumulate in proportion to cell volume. During mitosis, the total numbers of type 1 node proteins (cell cycle kinases Cdr1p, Cdr2p, Wee1p, and anillin Mid1p) are constant even when the nodes disassemble. Quantitative measurements provide strong evidence that both types of nodes have defined sizes and numbers of constituent proteins, as observed for cytokinesis nodes. Type 1 nodes assemble in two phases—a burst at the end of mitosis, followed by steady increase during interphase to double the initial number. Type 2 nodes containing Blt1p, Rho-GEF Gef2p, and kinesin Klp8p remain intact throughout the cell cycle and are constituents of the contractile ring. They are released from the contractile ring as it disassembles and then associate with type 1 nodes around the equator of the cell during interphase. PMID:28539404

GNOM regulates root hydrotropism and phototropism independently of PIN-mediated auxin transport.

PubMed

Moriwaki, Teppei; Miyazawa, Yutaka; Fujii, Nobuharu; Takahashi, Hideyuki

2014-02-01

Plant roots exhibit tropisms in response to gravity, unilateral light and moisture gradients. During gravitropism, an auxin gradient is established by PIN auxin transporters, leading to asymmetric growth. GNOM, a guanine nucleotide exchange factor of ARF GTPase (ARF-GEF), regulates PIN localization by regulating subcellular trafficking of PINs. Therefore, GNOM is important for gravitropism. We previously isolated mizu-kussei2 (miz2), which lacks hydrotropic responses; MIZ2 is allelic to GNOM. Since PIN proteins are not required for root hydrotropism in Arabidopsis, the role of GNOM in root hydrotropism should differ from that in gravitropism. To examine this possibility, we conducted genetic analysis of gnom(miz2) and gnom trans-heterozygotes. The mutant gnom(miz2), which lacks hydrotropic responses, was partially recovered by gnom(emb30-1), which lacks GEF activity, but not by gnom(B4049), which lacks heterotypic domain interactions. Furthermore, the phototropic response of gnom trans-heterozygotes differed from that of the pin2 mutant allele eir1-1. Moreover, defects in the polarities of PIN2 and auxin distribution in a severe gnom mutant were recovered by gnom(miz2). Therefore, an unknown GNOM-mediated vesicle trafficking system may mediate root hydrotropism and phototropism independently of PIN trafficking. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

G protein signaling in the parasite Entamoeba histolytica

PubMed Central

Bosch, Dustin E; Siderovski, David P

2013-01-01

The parasite Entamoeba histolytica causes amebic colitis and systemic amebiasis. Among the known amebic factors contributing to pathogenesis are signaling pathways involving heterotrimeric and Ras superfamily G proteins. Here, we review the current knowledge of the roles of heterotrimeric G protein subunits, Ras, Rho and Rab GTPase families in E. histolytica pathogenesis, as well as of their downstream signaling effectors and nucleotide cycle regulators. Heterotrimeric G protein signaling likely modulates amebic motility and attachment to and killing of host cells, in part through activation of an RGS-RhoGEF (regulator of G protein signaling–Rho guanine nucleotide exchange factor) effector. Rho family GTPases, as well as RhoGEFs and Rho effectors (formins and p21-activated kinases) regulate the dynamic actin cytoskeleton of E. histolytica and associated pathogenesis-related cellular processes, such as migration, invasion, phagocytosis and evasion of the host immune response by surface receptor capping. A remarkably large family of 91 Rab GTPases has multiple roles in a complex amebic vesicular trafficking system required for phagocytosis and pinocytosis and secretion of known virulence factors, such as amebapores and cysteine proteases. Although much remains to be discovered, recent studies of G protein signaling in E. histolytica have enhanced our understanding of parasitic pathogenesis and have also highlighted possible targets for pharmacological manipulation. PMID:23519208

Pak and Rac GTPases promote oncogenic KIT–induced neoplasms

PubMed Central

Martin, Holly; Mali, Raghuveer Singh; Ma, Peilin; Chatterjee, Anindya; Ramdas, Baskar; Sims, Emily; Munugalavadla, Veerendra; Ghosh, Joydeep; Mattingly, Ray R.; Visconte, Valeria; Tiu, Ramon V.; Vlaar, Cornelis P.; Dharmawardhane, Suranganie; Kapur, Reuben

2013-01-01

An acquired somatic mutation at codon 816 in the KIT receptor tyrosine kinase is associated with poor prognosis in patients with systemic mastocytosis and acute myeloid leukemia (AML). Treatment of leukemic cells bearing this mutation with an allosteric inhibitor of p21–activated kinase (Pak) or its genetic inactivation results in growth repression due to enhanced apoptosis. Inhibition of the upstream effector Rac abrogates the oncogene-induced growth and activity of Pak. Although both Rac1 and Rac2 are constitutively activated via the guanine nucleotide exchange factor (GEF) Vav1, loss of Rac1 or Rac2 alone moderately corrected the growth of KIT-bearing leukemic cells, whereas the combined loss resulted in 75% growth repression. In vivo, the inhibition of Vav or Rac or Pak delayed the onset of myeloproliferative neoplasms (MPNs) and corrected the associated pathology in mice. To assess the role of Rac GEFs in oncogene-induced transformation, we used an inhibitor of Rac, EHop-016, which specifically targets Vav1 and found that EHop-016 was a potent inhibitor of human and murine leukemic cell growth. These studies identify Pak and Rac GTPases, including Vav1, as potential therapeutic targets in MPN and AML involving an oncogenic form of KIT. PMID:24091327

The RhoGAP activity of CYK-4/MgcRacGAP functions non-canonically by promoting RhoA activation during cytokinesis

PubMed Central

Zhang, Donglei; Glotzer, Michael

2015-01-01

Cytokinesis requires activation of the GTPase RhoA. ECT-2, the exchange factor responsible for RhoA activation, is regulated to ensure spatiotemporal control of contractile ring assembly. Centralspindlin, composed of the Rho family GTPase-activating protein (RhoGAP) MgcRacGAP/CYK-4 and the kinesin MKLP1/ZEN-4, is known to activate ECT-2, but the underlying mechanism is not understood. We report that ECT-2-mediated RhoA activation depends on the ability of CYK-4 to localize to the plasma membrane, bind RhoA, and promote GTP hydrolysis by RhoA. Defects resulting from loss of CYK-4 RhoGAP activity can be rescued by activating mutations in ECT-2 or depletion of RGA-3/4, which functions as a conventional RhoGAP for RhoA. Consistent with CYK-4 RhoGAP activity contributing to GEF activation, the catalytic domains of CYK-4 and ECT-2 directly interact. Thus, counterintuitively, CYK-4 RhoGAP activity promotes RhoA activation. We propose that the most active form of the cytokinetic RhoGEF involves complex formation between ECT-2, centralspindlin and RhoA. DOI: http://dx.doi.org/10.7554/eLife.08898.001 PMID:26252513

The flipflop orphan genes are required for limb bud eversion in the Tribolium embryo.

PubMed

Thümecke, Susanne; Beermann, Anke; Klingler, Martin; Schröder, Reinhard

2017-01-01

Unlike Drosophila but similar to other arthropod and vertebrate embryos, the flour beetle Tribolium castaneum develops everted limb buds during embryogenesis. However, the molecular processes directing the evagination of epithelia are only poorly understood. Here we show that the newly discovered genes Tc-flipflop1 and Tc-flipflop2 are involved in regulating the directional budding of appendages. RNAi-knockdown of Tc-flipflop results in a variety of phenotypic traits. Most prominently, embryonic limb buds frequently grow inwards rather than out, leading to the development of inverted appendages inside the larval body. Moreover, affected embryos display dorsal closure defects. The Tc-flipflop genes are evolutionarily non-conserved, and their molecular function is not evident. We further found that Tc-RhoGEF2 , a highly-conserved gene known to be involved in actomyosin-dependent cell movement and cell shape changes, shows a Tc-flipflop -like RNAi-phenotype. The similarity of the inverted appendage phenotype in both the flipflop - and the RhoGEF2 RNAi gene knockdown led us to conclude that the Tc-flipflop orphan genes act in a Rho-dependent pathway that is essential for the early morphogenesis of polarised epithelial movements. Our work describes one of the few examples of an orphan gene playing a crucial role in an important developmental process.

Structural basis for the recognition of Asef by adenomatous polyposis coli

PubMed Central

Zhang, Zhenyi; Chen, Leyi; Gao, Lei; Lin, Kui; Zhu, Liang; Lu, Yang; Shi, Xiaoshan; Gao, Yuan; Zhou, Jing; Xu, Ping; Zhang, Jian; Wu, Geng

2012-01-01

Adenomatous polyposis coli (APC) regulates cell-cell adhesion and cell migration through activating the APC-stimulated guanine nucleotide-exchange factor (GEF; Asef), which is usually autoinhibited through the binding between its Src homology 3 (SH3) and Dbl homology (DH) domains. The APC-activated Asef stimulates the small GTPase Cdc42, which leads to decreased cell-cell adherence and enhanced cell migration. In colorectal cancers, truncated APC constitutively activates Asef and promotes cancer cell migration and angiogenesis. Here, we report crystal structures of the human APC/Asef complex. We find that the armadillo repeat domain of APC uses a highly conserved surface groove to recognize the APC-binding region (ABR) of Asef, conformation of which changes dramatically upon binding to APC. Key residues on APC and Asef for the complex formation were mutated and their importance was demonstrated by binding and activity assays. Structural superimposition of the APC/Asef complex with autoinhibited Asef suggests that the binding between APC and Asef might create a steric clash between Asef-DH domain and APC, which possibly leads to a conformational change in Asef that stimulates its GEF activity. Our structures thus elucidate the molecular mechanism of Asef recognition by APC, as well as provide a potential target for pharmaceutical intervention against cancers. PMID:21788986

Mutation-Specific Mechanisms of Hyperactivation of Noonan Syndrome SOS Molecules Detected with Single-molecule Imaging in Living Cells.

PubMed

Nakamura, Yuki; Umeki, Nobuhisa; Abe, Mitsuhiro; Sako, Yasushi

2017-10-26

Noonan syndrome (NS) is a congenital hereditary disorder associated with developmental and cardiac defects. Some patients with NS carry mutations in SOS, a guanine nucleotide exchange factor (GEF) for the small GTPase RAS. NS mutations have been identified not only in the GEF domain, but also in various domains of SOS, suggesting that multiple mechanisms disrupt SOS function. In this study, we examined three NS mutations in different domains of SOS to clarify the abnormality in its translocation to the plasma membrane, where SOS activates RAS. The association and dissociation kinetics between SOS tagged with a fluorescent protein and the living cell surface were observed in single molecules. All three mutants showed increased affinity for the plasma membrane, inducing excessive RAS signalling. However, the mechanisms by which their affinity was increased were specific to each mutant. Conformational disorder in the resting state, increased probability of a conformational change on the plasma membrane, and an increased association rate constant with the membrane receptor are the suggested mechanisms. These different properties cause the specific phenotypes of the mutants, which should be rescuable with different therapeutic strategies. Therefore, single-molecule kinetic analyses of living cells are useful for the pathological analysis of genetic diseases.

Rho proteins of plants--functional cycle and regulation of cytoskeletal dynamics.

PubMed

Mucha, Elena; Fricke, Inka; Schaefer, Antje; Wittinghofer, Alfred; Berken, Antje

2011-11-01

Rho-related ROP proteins are molecular switches that essentially regulate a wide variety of processes. Of central interest is their influence on the plant cytoskeleton by which they affect vital processes like cell division, growth, morphogenesis, and pathogen defense. ROPs switch between GTP- and GDP-bound conformations by strictly regulated nucleotide exchange and GTP-hydrolysis, and only the active GTP-form interacts with downstream effectors to ultimately provoke a biological response. However, the mode of action of the engaged regulators and effectors as well as their upstream and downstream interaction partners have long been largely unknown. As opposed to analogous systems in animals and fungi, plants use specific GTPase activating proteins (RopGAPs) with a unique domain composition and novel guanine nucleotide exchange factors (RopGEFs) with a probable link to cell surface receptors. Moreover, plants comprise novel effector molecules and adapters connecting ROPs to mostly unknown downstream targets on the route to the cytoskeleton. This review aims to summarize recent knowledge on the molecular mechanisms and reaction cascades involved in ROP dependent cytoskeletal rearrangements, addressing the structure and function of the unusual RopGAPs, RopGEFs and effectors, and the upstream and downstream pathways linking ROPs to cell receptor-like kinases, actin filaments, and microtubules. Copyright © 2010 Elsevier GmbH. All rights reserved.

45 CFR 1304.53 - Facilities, materials, and equipment.

Code of Federal Regulations, 2010 CFR

2010-10-01

.... (a) Head Start physical environment and facilities. (1) Grantee and delegate agencies must provide a physical environment and facilities conducive to learning and reflective of the different stages of... delegate agencies must provide a center-based environment free of toxins, such as cigarette smoke, lead...

Comparison of a traditional and non-traditional residential care facility for persons living with dementia and the impact of the environment on occupational engagement.

PubMed

Richards, Kieva; D'Cruz, Rachel; Harman, Suzanne; Stagnitti, Karen

2015-12-01

Dementia residential facilities can be described as traditional or non-traditional facilities. Non-traditional facilities aim to utilise principles of environmental design to create a milieu that supports persons experiencing cognitive decline. This study aimed to compare these two environments in rural Australia, and their influence on residents' occupational engagement. The Residential Environment Impact Survey (REIS) was used and consists of: a walk-through of the facility; activity observation; interviews with residents and employees. Thirteen residents were observed and four employees interviewed. Resident interviews did not occur given the population diagnosis of moderate to severe dementia. Descriptive data from the walk-through and activity observation were analysed for potential opportunities of occupational engagement. Interviews were thematically analysed to discern perception of occupational engagement of residents within their facility. Both facilities provided opportunities for occupational engagement. However, the non-traditional facility provided additional opportunities through employee interactions and features of the physical environment. Interviews revealed six themes: Comfortable environment; roles and responsibilities; getting to know the resident; more stimulation can elicit increased engagement; the home-like experience and environmental layout. These themes coupled with the features of the environment provided insight into the complexity of occupational engagement within this population. This study emphasises the influence of the physical and social environment on occupational engagement opportunities. A non-traditional dementia facility maximises these opportunities and can support development of best-practice guidelines within this population. © 2015 Occupational Therapy Australia.

Investigating walking environments in and around assisted living facilities: a facility visit study.

PubMed

Lu, Zhipeng

2010-01-01

This study explores assisted living residents' walking behaviors, locations where residents prefer to walk, and walking environments in and around assisted living facilities. Regular walking is beneficial to older adults' physical and psychological health. Yet frail older residents in assisted living are usually too sedentary to achieve these benefits. The physical environment plays an important role in promoting physical activity. However, there is little research exploring this relationship in assisted living settings. The researcher visited 34 assisted living facilities in a major Texas city. Methods included walk-through observation with the Assisted Living Facility Walking Environment Checklist, and interviews with administrators by open- and close-ended questions. The data from 26 facilities were analyzed using descriptive statistics (for quantitative data) and content analysis (for qualitative data). The results indicate that (a) residents were walking both indoors and outdoors for exercise or other purposes (e.g., going to destinations); (b) assisted living facility planning and design details-such as neighborhood sidewalk conditions, facility site selection, availability of seating, walking path configuration (e.g., looped/nonlooped path), amount of shading along the path, presence of handrails, existence of signage, etc.-may influence residents' walking behaviors; and (c) current assisted living facilities need improvement in all aspects to make their environments more walkable for residents. Findings of the study provide recommendations for assisted living facilities to improve the walkability of environments and to create environmental interventions to promote regular walking among their residents. This study also implies several directions for future research.

40 CFR 792.43 - Test system care facilities.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 31 2010-07-01 2010-07-01 true Test system care facilities. 792.43 Section 792.43 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) TOXIC SUBSTANCES CONTROL ACT (CONTINUED) GOOD LABORATORY PRACTICE STANDARDS Facilities § 792.43 Test system care facilities...

Regulation of Cdc42/Rac Signaling in the Establishment of Cell Polarity and Control of Cell Motility

DTIC Science & Technology

2004-08-01

Irazoqui Breast Cancer Predoctoral Traineeship Final Report Introduction Cdc42p, together with other polarity proteins, becomes polarized to a cap... Cancer Biology, Certificate in Cell and Molecular Biology. "* Awarded the Jane Coffin Childs Postdoctoral Fellowship for continuing research in the...Bemlp binds directly to both the Cdc42p-directed GEF Department of Pharmacology and Cancer Biology Duke University Medical Center, Durham, NC 27710

Relating Operational Art to the National Guard State Partnership Program

DTIC Science & Technology

2014-05-15

Program Coordinator GEF Global Employment of the Force DSCA Defense Support of Civil Authorities BCA Budget Control Act NATO North Atlantic Treaty...between Florida and the U.S. Virgin Islands are referred to as the Regional Security System (RSS). Florida and the U.S. Virgin Islands partner with... Atlantic Treaty Organization (NATO) in 2004 due, in part, to the mentorship from the Maryland National Guard.54 Not only did the Maryland National Guard

A facility for training Space Station astronauts

NASA Technical Reports Server (NTRS)

Hajare, Ankur R.; Schmidt, James R.

1992-01-01

The Space Station Training Facility (SSTF) will be the primary facility for training the Space Station Freedom astronauts and the Space Station Control Center ground support personnel. Conceptually, the SSTF will consist of two parts: a Student Environment and an Author Environment. The Student Environment will contain trainers, instructor stations, computers and other equipment necessary for training. The Author Environment will contain the systems that will be used to manage, develop, integrate, test and verify, operate and maintain the equipment and software in the Student Environment.

40 CFR 160.47 - Facilities for handling test, control, and reference substances.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Facilities for handling test, control, and reference substances. 160.47 Section 160.47 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) PESTICIDE PROGRAMS GOOD LABORATORY PRACTICE STANDARDS Facilities § 160.47 Facilities...

40 CFR 160.51 - Specimen and data storage facilities.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Specimen and data storage facilities. 160.51 Section 160.51 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) PESTICIDE PROGRAMS GOOD LABORATORY PRACTICE STANDARDS Facilities § 160.51 Specimen and data storage facilities. Space...

40 CFR 60.32c - Designated facilities.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 6 2010-07-01 2010-07-01 false Designated facilities. 60.32c Section 60.32c Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED... Solid Waste Landfills § 60.32c Designated facilities. (a) The designated facility to which the...

Design Process Overview

DOE Office of Scientific and Technical Information (OSTI.GOV)

Halkjaer-Knudsen, Vibeke

2014-11-01

For the purposes of this paper, a Biocontainment facility is a laboratory, production facility, or similar building that handles contagious biological materials in a safe and responsible manner. This specialized facility, also called a containment facility or a high containment facility reduces the potential for biological agents to be released into the environment, provides a safe work environment for the employees, and supports good laboratory practices.

The Impact of School Facilities on the Learning Environment

ERIC Educational Resources Information Center

Vandiver, Bert

2011-01-01

The purpose of this mixed methods study was to examine the impact of the quality of facilities on the educational environment in high schools located in northeast Texas. The intent of this research study was to determine the relationship between school facilities and the school-learning environment. This study was a mixed method research that used…

Dual system to reinforce biological containment of recombinant bacteria designed for rhizoremediation.

PubMed

Ronchel, M C; Ramos, J L

2001-06-01

Active biological containment (ABC) systems have been designed to control at will the survival or death of a bacterial population. These systems are based on the use of a killing gene, e.g., a porin-inducing protein such as the one encoded by the Escherichia coli gef gene, and a regulatory circuit that controls expression of the killing gene in response to the presence or absence of environmental signals. An ABC system for recombinant microorganisms that degrade a model pollutant was designed on the basis of the Pseudomonas putida TOL plasmid meta-cleavage regulatory circuit. The system consists of a fusion of the Pm promoter to lacI, whose expression is controlled by XylS with 3-methylbenzoate, and a fusion of a synthetic P(lac) promoter to gef. In the presence of the model pollutant, bacterial cells survived and degraded the target compound, whereas in the absence of the aromatic carboxylic acid cell death was induced. The system had two main drawbacks: (i) the slow death of the bacterial cells in soil versus the fast killing rate in liquid cultures in laboratory assays, and (ii) the appearance of mutants, at a rate of about 10(-8) per cell and generation, that did not die after the pollutant had been exhausted. We reinforced the ABC system by including it in a Deltaasd P. putida background. A P. putida Deltaasd mutant is viable only in complex medium supplemented with diaminopimelic acid, methionine, lysine, and threonine. We constructed a P. putida Deltaasd strain, called MCR7, with a Pm::asd fusion in the host chromosome. This strain was viable in the presence of 3-methylbenzoate because synthesis of the essential metabolites was achieved through XylS-dependent induction. In the P. putida MCR7 strain, an ABC system (Pm::lacI, xylS, P(lac)::gef) was incorporated into the host chromosome to yield strain MCR8. The number of MCR8 mutants that escaped killing was below our detection limit (<10(-9) mutants per cell and generation). The MCR8 strain survived and colonized rhizosphere soil with 3-methylbenzoate at a level similar to that of the wild-type strain. However, it disappeared in less than 20 to 25 days in soils without the pollutant, whereas an asd(+), biologically contained counterpart such as P. putida CMC4 was still detectable in soils after 100 days.

Incorporating Medium-Range Weather Forecasts in Seasonal Crop Scenarios over the Greater Horn of Africa to Support National/Regional/Local Decision Makers

NASA Astrophysics Data System (ADS)

Shukla, S.; Husak, G. J.; Funk, C. C.; Verdin, J. P.

2015-12-01

The USAID's Famine Early Warning Systems Network (FEWS NET) provides seasonal assessments of crop conditions over the Greater Horn of Africa (GHA) and other food insecure regions. These assessments and current livelihood, nutrition, market conditions and conflicts are used to generate food security scenarios that help national, regional and local decision makers target their resources and mitigate socio-economic losses. Among the various tools that FEWS NET uses is the FAO's Water Requirement Satisfaction Index (WRSI). The WRSI is a simple yet powerful crop assessment model that incorporates current moisture conditions (at the time of the issuance of forecast), precipitation scenarios, potential evapotranspiration and crop parameters to categorize crop conditions into different classes ranging from "failure" to "very good". The WRSI tool has been shown to have a good agreement with local crop yields in the GHA region. At present, the precipitation scenarios used to drive the WRSI are based on either a climatological forecast (that assigns equal chances of occurrence to all possible scenarios and has no skill over the forecast period) or a sea-surface temperature anomaly based scenario (which at best have skill at the seasonal scale). In both cases, the scenarios fail to capture the skill that can be attained by initial atmospheric conditions (i.e., medium-range weather forecasts). During the middle of a cropping season, when a week or two of poor rains can have a devastating effect, two weeks worth of skillful precipitation forecasts could improve the skill of the crop scenarios. With this working hypothesis, we examine the value of incorporating medium-range weather forecasts in improving the skill of crop scenarios in the GHA region. We use the NCEP's Global Ensemble Forecast system (GEFS) weather forecasts and examine the skill of crop scenarios generated using the GEFS weather forecasts with respect to the scenarios based solely on the climatological forecast. The period of analysis is from 1985-2010 (over which the reforecasts of GEFS is available) and the focus season is October-November-December. We examine the improvement (if any) in long-term skill, and present results for several recent drought events in the region.

Comparing the nutrition environment and practices of home- and centre-based child-care facilities.

PubMed

Martyniuk, Olivia J M; Vanderloo, Leigh M; Irwin, Jennifer D; Burke, Shauna M; Tucker, Patricia

2016-03-01

To assess and compare the nutrition environment and practices (as they relate to pre-schoolers) of centre- and home-based child-care facilities. Using a cross-sectional study design, nineteen child-care facilities (ten centre-based, nine home-based) were assessed for one full day using the Environment and Policy Assessment and Observation (EPAO) tool (consisting of a day-long observation/review of the nutrition environment, practices and related documents). Specifically, eight nutrition-related subscales were considered. Child-care facilities in London, Ontario, Canada. Child-care facilities were recruited through directors at centre-based programmes and the providers of home-based programmes. The mean total nutrition environment EPAO scores for centre- and home-based facilities were 12·3 (sd 1·94) and 10·8 (sd 0·78) out of 20 (where a higher score indicates a more supportive environment with regard to nutrition), respectively. The difference between the total nutrition environment EPAO score for centre- and home-based facilities was approaching significance (P=0·055). For both types of facilities, the highest nutrition subscale score (out of 20) was achieved in the staff behaviours domain (centre mean=17·4; home mean=17·0) and the lowest was in the nutrition training and education domain (centre mean=3·6; home mean=2·0). Additional research is needed to confirm these findings. In order to better support child-care staff and enhance the overall nutrition environment in child care, modifications to food practices could be adopted. Specifically, the nutritional quality of foods/beverages provided to pre-schoolers could be improved, nutrition-related training for child-care staff could be provided, and a nutrition curriculum could be created to educate pre-schoolers about healthy food choices.

Energy Optimization Assessment at U.S. Army Installations: West Point Military Academy, NY

DTIC Science & Technology

2008-09-01

Richland, WA 99352 Roland Ziegler GEF Final Report Approved for public release; distribution is unlimited. Prepared for U.S. Army Corps of Engineers ...Executive Order 13123 and Energy Policy Act (EPAct) 2005. A team of researchers from the Engineer Research and Development Cen- ter, Construction...CEP #5 Longer use of the backpressure steam turbine by increasing the low pres- sure steam demand 1,200,000 0 $150,000 0 $— $— $150,000

Identification of protein expression alterations in gefitinib-resistant human lung adenocarcinoma: PCNT and mPR play key roles in the development of gefitinib-associated resistance

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lin, Chi-Chen; Institute of Biomedical Science, and Rong Hsing Research Center for Translational Medicine, National Chung Hsing University, Taiwan; Department of Medical Research and Education, Taichung Veterans General Hospital, Taichung, Taiwan

2015-11-01

Gefitinib is the first-line chemotherapeutic drug for treating non-small cell lung cancer (NSCLC), which comprises nearly 85% of all lung cancer cases worldwide. However, most patients eventually develop drug resistance after 12–18 months of treatment. Hence, investigating the drug resistance mechanism and resistance-associated biomarkers is necessary. Two lung adenocarcinoma cell lines, PC9 and gefitinib-resistant PC9/Gef, were established for examining resistance mechanisms and identifying potential therapeutic targets. Two-dimensional differential gel electrophoresis and matrix-assisted laser desorption ionization time-of-flight mass spectrometry were used for examining global protein expression changes between PC9 and PC9/Gef. The results revealed that 164 identified proteins were associated withmore » the formation of gefitinib resistance in PC9 cells. Additional studies using RNA interference showed that progesterone receptor membrane component 1 and pericentrin proteins have major roles in gefitinib resistance. In conclusion, the proteomic approach enabled identifying of numerous proteins involved in gefitinib resistance. The results provide useful diagnostic markers and therapeutic candidates for treating gefitinib-resistant NSCLC. - Highlights: • 164 proteins associated with gefitinib resistance were identified through proteomic analysis. • In this study, a lung adenocarcinoma and its gefitinib resistant partner were established. • mPR and PCNT proteins have evidenced to play important roles in gefitinib resistance.« less

PI3K regulates endocytosis after insulin secretion by mediating signaling crosstalk between Arf6 and Rab27a.

PubMed

Yamaoka, Mami; Ando, Tomomi; Terabayashi, Takeshi; Okamoto, Mitsuhiro; Takei, Masahiro; Nishioka, Tomoki; Kaibuchi, Kozo; Matsunaga, Kohichi; Ishizaki, Ray; Izumi, Tetsuro; Niki, Ichiro; Ishizaki, Toshimasa; Kimura, Toshihide

2016-02-01

In secretory cells, endocytosis is coupled to exocytosis to enable proper secretion. Although endocytosis is crucial to maintain cellular homeostasis before and after secretion, knowledge about secretagogue-induced endocytosis in secretory cells is still limited. Here, we searched for proteins that interacted with the Rab27a GTPase-activating protein (GAP) EPI64 (also known as TBC1D10A) and identified the Arf6 guanine-nucleotide-exchange factor (GEF) ARNO (also known as CYTH2) in pancreatic β-cells. We found that the insulin secretagogue glucose promotes phosphatidylinositol (3,4,5)-trisphosphate (PIP3) generation through phosphoinositide 3-kinase (PI3K), thereby recruiting ARNO to the intracellular side of the plasma membrane. Peripheral ARNO promotes clathrin assembly through its GEF activity for Arf6 and regulates the early stage of endocytosis. We also found that peripheral ARNO recruits EPI64 to the same area and that the interaction requires glucose-induced endocytosis in pancreatic β-cells. Given that GTP- and GDP-bound Rab27a regulate exocytosis and the late stage of endocytosis, our results indicate that the glucose-induced activation of PI3K plays a pivotal role in exocytosis-endocytosis coupling, and that ARNO and EPI64 regulate endocytosis at distinct stages. © 2016. Published by The Company of Biologists Ltd.

The Arf GEF GBF1 and Arf4 synergize with the sensory receptor cargo, rhodopsin, to regulate ciliary membrane trafficking.

PubMed

Wang, Jing; Fresquez, Theresa; Kandachar, Vasundhara; Deretic, Dusanka

2017-12-01

The small GTPase Arf4 and the Arf GTPase-activating protein (GAP) ASAP1 cooperatively sequester sensory receptor cargo into transport carriers targeted to primary cilia, but the input that drives Arf4 activation in this process remains unknown. Here, we show, by using frog retinas and recombinant human proteins, that during the carrier biogenesis from the photoreceptor Golgi/ trans -Golgi network (TGN) a functional complex is formed between Arf4, the Arf guanine nucleotide exchange factor (GEF) GBF1 and the light-sensing receptor, rhodopsin. Rhodopsin and Arf4 bind the regulatory N-terminal dimerization and cyclophillin-binding (DCB)-homology upstream of Sec7 (HUS) domain of GBF1. The complex is sensitive to Golgicide A (GCA), a selective inhibitor of GBF1 that accordingly blocks rhodopsin delivery to the cilia, without disrupting the photoreceptor Golgi. The emergence of newly synthesized rhodopsin in the endomembrane system is essential for GBF1-Arf4 complex formation in vivo Notably, GBF1 interacts with the Arf GAP ASAP1 in a GCA-resistant manner. Our findings indicate that converging signals on GBF1 from the influx of cargo into the Golgi/TGN and the feedback from Arf4, combined with input from ASAP1, control Arf4 activation during sensory membrane trafficking to primary cilia. © 2017. Published by The Company of Biologists Ltd.

BLOC-3 Mutated in Hermansky-Pudlak Syndrome Is a Rab32/38 Guanine Nucleotide Exchange Factor

PubMed Central

Gerondopoulos, Andreas; Langemeyer, Lars; Liang, Jin-Rui; Linford, Andrea; Barr, Francis A.

2012-01-01

Summary Hermansky-Pudlak syndrome (HPS) is a human disease characterized by partial loss of pigmentation and impaired blood clotting [1–3]. These symptoms are caused by defects in the biogenesis of melanosomes and platelet dense granules, often referred to as lysosome-related organelles [2]. Genes mutated in HPS encode subunits of the biogenesis of lysosome-related organelles complexes (BLOCs). BLOC-1 and BLOC-2, together with the AP-3 clathrin adaptor complex, act at early endosomes to sort components required for melanin formation and melanosome biogenesis away from the degradative lysosomal pathway toward early stage melanosomes [4–6]. However the molecular functions of the Hps1-Hps4 complex BLOC-3 remain mysterious [7–9]. Like other trafficking pathways, melanosome biogenesis and transport of enzymes involved in pigmentation involves specific Rab GTPases, in this instance Rab32 and Rab38 [10–12]. We now demonstrate that BLOC-3 is a Rab32 and Rab38 guanine nucleotide exchange factor (GEF). Silencing of the BLOC-3 subunits Hps1 and Hps4 results in the mislocalization of Rab32 and Rab38 and reduction in pigmentation. In addition, we show that BLOC-3 can promote specific membrane recruitment of Rab32/38. BLOC-3 therefore defines a novel Rab GEF family with a specific function in the biogenesis of lysosome-related organelles. PMID:23084991

Site-specific regulation of the GEF Cdc24p by the scaffold protein Far1p during yeast mating

PubMed Central

Wiget, Philippe; Shimada, Yukiko; Butty, Anne-Christine; Bi, Efrei; Peter, Matthias

2004-01-01

Receptor-mediated cell polarization via heterotrimeric G-proteins induces cytoskeletal rearrangements in a variety of organisms. In yeast, Far1p is required for orienting cell growth towards the mating partner by linking activated Gβγ to the guanine-nucleotide exchange factor (GEF) Cdc24p, which activates the Rho-type GTPase Cdc42p. Here we investigated the role of Far1p in the regulation of Cdc24p in vivo. Using time-lapse microscopy of mating cells and artificial membrane targeting of Far1p, we show that Far1p is necessary and sufficient to recruit Cdc24p to the plasma membrane. Wild-type Far1p contains a PH-like domain, which is required for its membrane localization in vivo. Interestingly, expression of membrane-targeted Far1p causes toxicity, most likely by activating Cdc42p uniformly at the cell cortex. The ability of full-length Far1p to function as an activator of Cdc24p in vivo requires its interaction with Cdc24p and Gβγ. Our results imply that Gβγ not only targets Far1p to the correct site but may also trigger a conformational change in Far1p that is required for its ability to activate Cdc24p in vivo. PMID:14988725

The Phosphatidylinositol (3,4,5)-Trisphosphate-dependent Rac Exchanger 1·Ras-related C3 Botulinum Toxin Substrate 1 (P-Rex1·Rac1) Complex Reveals the Basis of Rac1 Activation in Breast Cancer Cells.

PubMed

Lucato, Christina M; Halls, Michelle L; Ooms, Lisa M; Liu, Heng-Jia; Mitchell, Christina A; Whisstock, James C; Ellisdon, Andrew M

2015-08-21

The P-Rex (phosphatidylinositol (3,4,5)-trisphosphate (PIP3)-dependent Rac exchanger) family (P-Rex1 and P-Rex2) of the Rho guanine nucleotide exchange factors (Rho GEFs) activate Rac GTPases to regulate cell migration, invasion, and metastasis in several human cancers. The family is unique among Rho GEFs, as their activity is regulated by the synergistic binding of PIP3 and Gβγ at the plasma membrane. However, the molecular mechanism of this family of multi-domain proteins remains unclear. We report the 1.95 Å crystal structure of the catalytic P-Rex1 DH-PH tandem domain in complex with its cognate GTPase, Rac1 (Ras-related C3 botulinum toxin substrate-1). Mutations in the P-Rex1·Rac1 interface revealed a critical role for this complex in signaling downstream of receptor tyrosine kinases and G protein-coupled receptors. The structural data indicated that the PIP3/Gβγ binding sites are on the opposite surface and markedly removed from the Rac1 interface, supporting a model whereby P-Rex1 binding to PIP3 and/or Gβγ releases inhibitory C-terminal domains to expose the Rac1 binding site. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

Dbl oncogene expression in MCF-10 A epithelial cells disrupts mammary acinar architecture, induces EMT and angiogenic factor secretion

PubMed Central

Vanni, Cristina; Ognibene, Marzia; Finetti, Federica; Mancini, Patrizia; Cabodi, Sara; Segalerba, Daniela; Torrisi, Maria Rosaria; Donnini, Sandra; Bosco, Maria Carla; Varesio, Luigi; Eva, Alessandra

2015-01-01

The proteins of the Dbl family are guanine nucleotide exchange factors (GEFs) of Rho GTPases and are known to be involved in cell growth regulation. Alterations of the normal function of these proteins lead to pathological processes such as developmental disorders, neoplastic transformation, and tumor metastasis. We have previously demonstrated that expression of Dbl oncogene in lens epithelial cells modulates genes encoding proteins involved in epithelial-mesenchymal-transition (EMT) and induces angiogenesis in the lens. Our present study was undertaken to investigate the role of Dbl oncogene in epithelial cells transformation, providing new insights into carcinoma progression.To assess how Dbl oncogene can modulate EMT, cell migration, morphogenesis, and expression of pro-apoptotic and angiogenic factors we utilized bi- and 3-dimensional cultures of MCF-10 A cells. We show that upon Dbl expression MCF-10 A cells undergo EMT. In addition, we found that Dbl overexpression sustains Cdc42 and Rac activation inducing morphological alterations, characterized by the presence of lamellipodia and conferring a high migratory capacity to the cells. Moreover, Dbl expressing MCF-10 A cells form altered 3D structures and can induce angiogenesis by producing proangiogenic factors such as CCL2. These results support a role for Dbl oncogene in epithelial cell differentiation and transformation and suggest the relevance of GEF deregulation in tumor onset and progression. PMID:25723869

Acetylation of the RhoA GEF Net1A controls its subcellular localization and activity

PubMed Central

Song, Eun Hyeon; Oh, Wonkyung; Ulu, Arzu; Carr, Heather S.; Zuo, Yan; Frost, Jeffrey A.

2015-01-01

ABSTRACT Net1 isoform A (Net1A) is a RhoA GEF that is required for cell motility and invasion in multiple cancers. Nuclear localization of Net1A negatively regulates its activity, and we have recently shown that Rac1 stimulates Net1A relocalization to the plasma membrane to promote RhoA activation and cytoskeletal reorganization. However, mechanisms controlling the subcellular localization of Net1A are not well understood. Here, we show that Net1A contains two nuclear localization signal (NLS) sequences within its N-terminus and that residues surrounding the second NLS sequence are acetylated. Treatment of cells with deacetylase inhibitors or expression of active Rac1 promotes Net1A acetylation. Deacetylase inhibition is sufficient for Net1A relocalization outside the nucleus, and replacement of the N-terminal acetylation sites with arginine residues prevents cytoplasmic accumulation of Net1A caused by deacetylase inhibition or EGF stimulation. By contrast, replacement of these sites with glutamine residues is sufficient for Net1A relocalization, RhoA activation and downstream signaling. Moreover, the N-terminal acetylation sites are required for rescue of F-actin accumulation and focal adhesion maturation in Net1 knockout MEFs. These data indicate that Net1A acetylation regulates its subcellular localization to impact on RhoA activity and actin cytoskeletal organization. PMID:25588829

Rabex-5 ubiquitin ligase activity restricts Ras signaling to establish pathway homeostasis in Drosophila.

PubMed

Yan, Hua; Jahanshahi, Maryam; Horvath, Elizabeth A; Liu, Hsiu-Yu; Pfleger, Cathie M

2010-08-10

The Ras signaling pathway allows cells to translate external cues into diverse biological responses. Depending on context and the threshold reached, Ras signaling can promote growth, proliferation, differentiation, or cell survival. Failure to maintain precise control of Ras can have adverse physiological consequences. Indeed, excess Ras signaling disrupts developmental patterning and causes developmental disorders [1, 2], and in mature tissues, it can lead to cancer [3-5]. We identify Rabex-5 as a new component of Ras signaling crucial for achieving proper pathway outputs in multiple contexts in vivo. We show that Drosophila Rabex-5 restricts Ras signaling to establish organism size, wing vein pattern, and eye versus antennal fate. Rabex-5 has both Rab5 guanine nucleotide exchange factor (GEF) activity that regulates endocytic trafficking [6] and ubiquitin ligase activity [7, 8]. Surprisingly, overexpression studies demonstrate that Rabex-5 ubiquitin ligase activity, not its Rab5 GEF activity, is required to restrict wing vein specification and to suppress the eye phenotypes of oncogenic Ras expression. Furthermore, genetic interaction experiments indicate that Rabex-5 acts at the step of Ras, and tissue culture studies show that Rabex-5 promotes Ras ubiquitination. Together, these findings reveal a new mechanism for attenuating Ras signaling in vivo and suggest an important role for Rabex-5-mediated Ras ubiquitination in pathway homeostasis. Copyright (c) 2010 Elsevier Ltd. All rights reserved.

33 CFR 154.1040 - Specific requirements for facilities that could reasonably be expected to cause substantial harm...

Code of Federal Regulations, 2011 CFR

2011-07-01

... facilities that could reasonably be expected to cause substantial harm to the environment. 154.1040 Section... to the environment. (a) The owner or operator of a facility that, under § 154.1015, could reasonably be expected to cause substantial harm to the environment, shall submit a response plan that meets the...

33 CFR 154.1040 - Specific requirements for facilities that could reasonably be expected to cause substantial harm...

Code of Federal Regulations, 2012 CFR

2012-07-01

... facilities that could reasonably be expected to cause substantial harm to the environment. 154.1040 Section... to the environment. (a) The owner or operator of a facility that, under § 154.1015, could reasonably be expected to cause substantial harm to the environment, shall submit a response plan that meets the...

33 CFR 154.1040 - Specific requirements for facilities that could reasonably be expected to cause substantial harm...

Code of Federal Regulations, 2013 CFR

2013-07-01

... facilities that could reasonably be expected to cause substantial harm to the environment. 154.1040 Section... to the environment. (a) The owner or operator of a facility that, under § 154.1015, could reasonably be expected to cause substantial harm to the environment, shall submit a response plan that meets the...

33 CFR 154.1040 - Specific requirements for facilities that could reasonably be expected to cause substantial harm...

Code of Federal Regulations, 2014 CFR

2014-07-01

... facilities that could reasonably be expected to cause substantial harm to the environment. 154.1040 Section... to the environment. (a) The owner or operator of a facility that, under § 154.1015, could reasonably be expected to cause substantial harm to the environment, shall submit a response plan that meets the...

33 CFR 154.1040 - Specific requirements for facilities that could reasonably be expected to cause substantial harm...

Code of Federal Regulations, 2010 CFR

2010-07-01

... facilities that could reasonably be expected to cause substantial harm to the environment. 154.1040 Section... to the environment. (a) The owner or operator of a facility that, under § 154.1015, could reasonably be expected to cause substantial harm to the environment, shall submit a response plan that meets the...

42 CFR 485.62 - Condition of participation: Physical environment.

Code of Federal Regulations, 2012 CFR

2012-10-01

... 42 Public Health 5 2012-10-01 2012-10-01 false Condition of participation: Physical environment... of participation: Physical environment. The facility must provide a physical environment that... patients. The physical premises of the facility and those areas of its surrounding physical structure that...

42 CFR 485.62 - Condition of participation: Physical environment.

Code of Federal Regulations, 2013 CFR

2013-10-01

... 42 Public Health 5 2013-10-01 2013-10-01 false Condition of participation: Physical environment... of participation: Physical environment. The facility must provide a physical environment that... patients. The physical premises of the facility and those areas of its surrounding physical structure that...

42 CFR 485.62 - Condition of participation: Physical environment.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 42 Public Health 5 2014-10-01 2014-10-01 false Condition of participation: Physical environment... of participation: Physical environment. The facility must provide a physical environment that... patients. The physical premises of the facility and those areas of its surrounding physical structure that...

40 CFR 270.300 - What container information must I keep at my facility?

Code of Federal Regulations, 2013 CFR

2013-07-01

... 40 Protection of Environment 28 2013-07-01 2013-07-01 false What container information must I keep at my facility? 270.300 Section 270.300 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY... Facility § 270.300 What container information must I keep at my facility? If you store or treat hazardous...

Who Delivers without Water? A Multi Country Analysis of Water and Sanitation in the Childbirth Environment.

PubMed

Gon, Giorgia; Restrepo-Méndez, María Clara; Campbell, Oona M R; Barros, Aluísio J D; Woodd, Susannah; Benova, Lenka; Graham, Wendy J

2016-01-01

Hygiene during childbirth is essential to the health of mothers and newborns, irrespective of where birth takes place. This paper investigates the status of water and sanitation in both the home and facility childbirth environments, and for whom and where this is a more significant problem. We used three datasets: a global dataset, with information on the home environment from 58 countries, and two datasets for each of four countries in Eastern Africa: a healthcare facility dataset, and a dataset that incorporated information on facilities and the home environment to create a comprehensive description of birth environments in those countries. We constructed indices of improved water, and improved water and sanitation combined (WATSAN), for the home and healthcare facilities. The Joint Monitoring Program was used to construct indices for household; we tailored them to the facility context-household and facility indices include different components. We described what proportion of women delivered in an environment with improved WATSAN. For those women who delivered at home, we calculated what proportion had improved WATSAN by socio-economic status, education and rural-urban status. Among women delivering at home (58 countries), coverage of improved WATSAN by region varied from 9% to 53%. Fewer than 15% of women who delivered at home in Sub-Saharan Africa, had access to water and sanitation infrastructure (range 0.1% to 37%). This was worse among the poorest, the less educated and those living in rural areas. In Eastern Africa, where we looked at both the home and facility childbirth environment, a third of women delivered in an environment with improved water in Uganda and Rwanda; whereas, 18% of women in Kenya and 7% in Tanzania delivered with improved water and sanitation. Across the four countries, less than half of the facility deliveries had improved water, or improved water and sanitation in the childbirth environment. Access to water and sanitation during childbirth is poor across low and middle-income countries. Even when women travel to health facilities for childbirth, they are not guaranteed access to basic WATSAN infrastructure. These indicators should be measured routinely in order to inform improvements.

The association between the physical environment and the well-being of older people in residential care facilities: A multilevel analysis.

PubMed

Nordin, Susanna; McKee, Kevin; Wijk, Helle; Elf, Marie

2017-12-01

To investigate the associations between the quality of the physical environment and the psychological and social well-being of older people living in residential care facilities. Many older people in care facilities have cognitive and physical frailties and are at risk of experiencing low levels of well-being. High-quality physical environments can support older people as frailty increases and promote their well-being. Although the importance of the physical environment for residents' well-being is recognized, more research is needed. A cross-sectional survey of 20 care facilities from each of which 10 residents were sampled. As the individual resident data were nested in the facilities, a multilevel analysis was conducted. Data were collected during 2013 and 2014. The care facilities were purposely sampled to ensure a high level of variation in their physical characteristics. Residents' demographic and health data were collected via medical records and interviews. Residents' well-being and perceived quality of care were assessed via questionnaires and interviews. Environmental quality was assessed with a structured observational instrument. Multilevel analysis indicated that cognitive support in the physical environment was associated with residents' social well-being, after controlling for independence and perceived care quality. However, no significant association was found between the physical environment and residents' psychological well-being. Our study demonstrates the role of the physical environment for enhancing the social well-being of frail older people. Professionals and practitioners involved in the design of care facilities have a responsibility to ensure that such facilities meet high-quality specifications. © 2017 The Authors. Journal of Advanced Nursing Published by John Wiley & Sons Ltd.

The physical environment of purpose-built and non-purpose-built supported housing for persons with psychiatric disabilities in Sweden.

PubMed

Johansson, Maria; Brunt, David

2012-04-01

The primary aim of the present study was to investigate if methods derived from environmental psychology can be used to study the qualities of the physical environment of supported housing facilities for persons with psychiatric disabilities. Three units of analysis were selected: the private area, the common indoor area, and the outdoor area. Expert assessments of 110 features of the physical environment in these units and semantic environmental description of the visual experience of them consistently showed that purpose-built supported housing facilities had more physical features important for high quality residential environments than the non-purpose-built supported housing facilities. The employed methods were thus seen to be able to describe and discriminate between qualities in the physical environment of supported housing facilities. Suggestions for the development of tools for the assessment of the physical environment in supported housing are made.

PELS (Planetary Environmental Liquid Simulator): a new type of simulation facility to study extraterrestrial aqueous environments.

PubMed

Martin, Derek; Cockell, Charles S

2015-02-01

Investigations of other planetary bodies, including Mars and icy moons such as Enceladus and Europa, show that they may have hosted aqueous environments in the past and may do so even today. Therefore, a major challenge in astrobiology is to build facilities that will allow us to study the geochemistry and habitability of these extraterrestrial environments. Here, we describe a simulation facility (PELS: Planetary Environmental Liquid Simulator) with the capability for liquid input and output that allows for the study of such environments. The facility, containing six separate sample vessels, allows for statistical replication of samples. Control of pressure, gas composition, UV irradiation conditions, and temperature allows for the precise replication of aqueous conditions, including subzero brines under martian atmospheric conditions. A sample acquisition system allows for the collection of both liquid and solid samples from within the chamber without breaking the atmospheric conditions, enabling detailed studies of the geochemical evolution and habitability of past and present extraterrestrial environments. The facility we describe represents a new frontier in planetary simulation-continuous flow-through simulation of extraterrestrial aqueous environments.

Regulation of ATM-Dependent DNA Damage Responses in Breast Cancer by the RhoGEF Net1

DTIC Science & Technology

2013-04-01

Science 279: 509-514. 5. Jaffe AB. et al., (2010) RhoGTPases: Biochemistry and Biology. Annu. Rev. Cell Dev. Biol. 21:247-269. 6. Rossman KL, et al...exchange factor Net1 is regulated by nuclear sequestration. J. Biol. Chem. 277:17, 14581-14588. 17. Harper JW, et al., (2007) The DNA Damage Response: Ten...Research (AACR) Annual Meeting and 2013 Annual Cancer Research Biochemistry Retreat Regulation of ATM-dependent DNA damage signaling in human breast

Adaptation to heavy rainfall events: watershed-community planning of soil and water conservation technologies in Syria

NASA Astrophysics Data System (ADS)

Ziadat, Feras; Al-Wadaey, Ahmed; Masri, Zuhair; Sakai, Hirokazu

2010-05-01

The Fourth Assessment Report of the Intergovernmental Panel on Climate Change (IPCC) and other research, predict a significant future increase in the frequency and intensity of heavy rainfall events in many regions. This increase runoff and soil erosion, and reduce agricultural productivity, as well as increasing risks of flood damage to crops and infrastructure. Implementing adaptation measures and improved land management through erosion control and soil protection are among those that protect water and agriculture and limit their vulnerability. Soil erosion control practices are often based on long-term climatic averages. Special attention is needed to provide protection against average high-return frequency storms as well as severe storms with low-return frequency. Suitable and affordable soil conservation plans, coupled with an appropriate enabling environment, are needed. A watershed and community were selected in the mountainous area of North West Syria. The fields represent the non-tropical highland dry areas and dominated by olive orchards on steep slopes. Farmers were aware of resource degradation and productivity reduction, but lacked financial capital to implement the needed adaptation measures. A micro-credit system was established with the help of the UNDP Global Environment Facility - Small Grants Program (GEF-SGP) with small grants available for each farmer. Haphazard implementation on scattered fields proved inefficient in demonstrating obvious impact. Therefore, each watershed was classified into three erosion risk categories (high, moderate and low), derived from maps of flow accumulation, slope steepness, slope shape and land use. Using field survey of land ownership, the boundaries of 168 farms in the watersheds were mapped. Farmers' fields were classified using the erosion-risk map and considering the on-farm erosion hazard and the off-farm effect on other farmers' fields following the hillslope sequence. More than 60% of the farms were classified into high erosion risk areas. Accordingly, a community-watershed plan was established and revised with the community committee. Loans to implement soil and water conservation measures were distributed to 52 farmers based on the priorities of their farms. Results from four runoff events in 2009 showed that one erosive runoff event can deliver more than 50% of the total soil loss. Implementing semi-circular bunds reduced rill erosion by 40% and captured 3.4 tons of sediments per hectare. The effect of this approach in limiting the negative impact of extreme rainfall events, at watershed and field levels, are now being quantified and modeled. Keywords: climate change, land use, soil erosion, GIS, flow accumulation, land tenure.

40 CFR 160.15 - Inspection of a testing facility.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Inspection of a testing facility. 160.15 Section 160.15 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) PESTICIDE PROGRAMS GOOD LABORATORY PRACTICE STANDARDS General Provisions § 160.15 Inspection of a testing facility...

40 CFR 792.47 - Facilities for handling test, control, and reference substances.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 31 2010-07-01 2010-07-01 true Facilities for handling test, control, and reference substances. 792.47 Section 792.47 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) TOXIC SUBSTANCES CONTROL ACT (CONTINUED) GOOD LABORATORY PRACTICE STANDARDS Facilities...

40 CFR 792.51 - Specimen and data storage facilities.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 31 2010-07-01 2010-07-01 true Specimen and data storage facilities. 792.51 Section 792.51 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) TOXIC SUBSTANCES CONTROL ACT (CONTINUED) GOOD LABORATORY PRACTICE STANDARDS Facilities § 792.51 Specimen and data...

40 CFR 256.42 - Recommendations for assuring facility development.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 25 2011-07-01 2011-07-01 false Recommendations for assuring facility development. 256.42 Section 256.42 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID... Planning and Implementation § 256.42 Recommendations for assuring facility development. (a) The State plan...

40 CFR 112.21 - Facility response training and drills/exercises.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 23 2012-07-01 2012-07-01 false Facility response training and drills/exercises. 112.21 Section 112.21 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS OIL POLLUTION PREVENTION Response Requirements § 112.21 Facility response training and drills...

40 CFR 112.21 - Facility response training and drills/exercises.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 22 2014-07-01 2013-07-01 true Facility response training and drills/exercises. 112.21 Section 112.21 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS OIL POLLUTION PREVENTION Response Requirements § 112.21 Facility response training and drills...

40 CFR 112.21 - Facility response training and drills/exercises.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 40 Protection of Environment 23 2013-07-01 2013-07-01 false Facility response training and drills/exercises. 112.21 Section 112.21 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS OIL POLLUTION PREVENTION Response Requirements § 112.21 Facility response training and drills...

40 CFR 112.21 - Facility response training and drills/exercises.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 21 2010-07-01 2010-07-01 false Facility response training and drills/exercises. 112.21 Section 112.21 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS OIL POLLUTION PREVENTION Response Requirements § 112.21 Facility response training and drills...

40 CFR 112.21 - Facility response training and drills/exercises.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 22 2011-07-01 2011-07-01 false Facility response training and drills/exercises. 112.21 Section 112.21 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS OIL POLLUTION PREVENTION Response Requirements § 112.21 Facility response training and drills...

40 CFR 60.300 - Applicability and designation of affected facility.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 7 2012-07-01 2012-07-01 false Applicability and designation of affected facility. 60.300 Section 60.300 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY... Performance for Grain Elevators § 60.300 Applicability and designation of affected facility. (a) The...

40 CFR 60.300 - Applicability and designation of affected facility.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 6 2011-07-01 2011-07-01 false Applicability and designation of affected facility. 60.300 Section 60.300 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY... Performance for Grain Elevators § 60.300 Applicability and designation of affected facility. (a) The...

40 CFR 792.45 - Test system supply facilities.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 31 2010-07-01 2010-07-01 true Test system supply facilities. 792.45 Section 792.45 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) TOXIC SUBSTANCES... facilities. (a) There shall be storage areas, as needed, for feed, nutrients, soils, bedding, supplies, and...

40 CFR 160.15 - Inspection of a testing facility.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 24 2011-07-01 2011-07-01 false Inspection of a testing facility. 160.15 Section 160.15 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) PESTICIDE PROGRAMS GOOD LABORATORY PRACTICE STANDARDS General Provisions § 160.15 Inspection of a testing facility...

Computational Tools and Facilities for the Next-Generation Analysis and Design Environment

NASA Technical Reports Server (NTRS)

Noor, Ahmed K. (Compiler); Malone, John B. (Compiler)

1997-01-01

This document contains presentations from the joint UVA/NASA Workshop on Computational Tools and Facilities for the Next-Generation Analysis and Design Environment held at the Virginia Consortium of Engineering and Science Universities in Hampton, Virginia on September 17-18, 1996. The presentations focused on the computational tools and facilities for analysis and design of engineering systems, including, real-time simulations, immersive systems, collaborative engineering environment, Web-based tools and interactive media for technical training. Workshop attendees represented NASA, commercial software developers, the aerospace industry, government labs, and academia. The workshop objectives were to assess the level of maturity of a number of computational tools and facilities and their potential for application to the next-generation integrated design environment.

40 CFR 256.41 - Recommendations for assessing the need for facilities.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 25 2011-07-01 2011-07-01 false Recommendations for assessing the need for facilities. 256.41 Section 256.41 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY... their impact on facility needs should be assessed. (3) Current and projected movement of solid and...

40 CFR 256.41 - Recommendations for assessing the need for facilities.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 24 2010-07-01 2010-07-01 false Recommendations for assessing the need for facilities. 256.41 Section 256.41 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY... their impact on facility needs should be assessed. (3) Current and projected movement of solid and...

40 CFR 255.33 - Inclusion of Federal facilities and Native American Reservations.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 40 Protection of Environment 26 2013-07-01 2013-07-01 false Inclusion of Federal facilities and Native American Reservations. 255.33 Section 255.33 Protection of Environment ENVIRONMENTAL PROTECTION... Responsibilities of Identified Agencies and Relationship to Other Programs § 255.33 Inclusion of Federal facilities...

40 CFR 255.33 - Inclusion of Federal facilities and Native American Reservations.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 26 2012-07-01 2011-07-01 true Inclusion of Federal facilities and Native American Reservations. 255.33 Section 255.33 Protection of Environment ENVIRONMENTAL PROTECTION... Responsibilities of Identified Agencies and Relationship to Other Programs § 255.33 Inclusion of Federal facilities...

40 CFR 255.33 - Inclusion of Federal facilities and Native American Reservations.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 25 2014-07-01 2014-07-01 false Inclusion of Federal facilities and Native American Reservations. 255.33 Section 255.33 Protection of Environment ENVIRONMENTAL PROTECTION... Responsibilities of Identified Agencies and Relationship to Other Programs § 255.33 Inclusion of Federal facilities...

40 CFR 255.33 - Inclusion of Federal facilities and Native American Reservations.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 24 2010-07-01 2010-07-01 false Inclusion of Federal facilities and Native American Reservations. 255.33 Section 255.33 Protection of Environment ENVIRONMENTAL PROTECTION... Responsibilities of Identified Agencies and Relationship to Other Programs § 255.33 Inclusion of Federal facilities...

40 CFR 255.33 - Inclusion of Federal facilities and Native American Reservations.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 25 2011-07-01 2011-07-01 false Inclusion of Federal facilities and Native American Reservations. 255.33 Section 255.33 Protection of Environment ENVIRONMENTAL PROTECTION... Responsibilities of Identified Agencies and Relationship to Other Programs § 255.33 Inclusion of Federal facilities...

Effects of a Facility Dog on Student Learning and Learning Environment

ERIC Educational Resources Information Center

Bradley, Jordana; Maldonado, Nancy

2013-01-01

Educators must consider alternative teaching strategies. Facility dogs as an instructional enhancement are an innovative teaching approach. This case study, guided by human-animal bond theory, investigated how the presence of a trained facility dog, Smooch, affected the school environment. Interviews, field notes and observations were used to…

40 CFR 112.20 - Facility response plans.

Code of Federal Regulations, 2012 CFR

2012-07-01

... environmental factors that the Regional Administrator determines to be relevant to protecting the environment... 40 Protection of Environment 23 2012-07-01 2012-07-01 false Facility response plans. 112.20 Section 112.20 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS OIL...

40 CFR 112.20 - Facility response plans.

Code of Federal Regulations, 2013 CFR

2013-07-01

... environmental factors that the Regional Administrator determines to be relevant to protecting the environment... 40 Protection of Environment 23 2013-07-01 2013-07-01 false Facility response plans. 112.20 Section 112.20 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS OIL...

40 CFR 112.20 - Facility response plans.

Code of Federal Regulations, 2014 CFR

2014-07-01

... environmental factors that the Regional Administrator determines to be relevant to protecting the environment... 40 Protection of Environment 22 2014-07-01 2013-07-01 true Facility response plans. 112.20 Section 112.20 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS OIL...

40 CFR 265.31 - Maintenance and operation of facility.

Code of Federal Regulations, 2010 CFR

2010-07-01

..., soil, or surface water which could threaten human health or the -environment. ... 40 Protection of Environment 25 2010-07-01 2010-07-01 false Maintenance and operation of facility. 265.31 Section 265.31 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID...

Aerospace Test Facilities at NASA LeRC Plumbrook

NASA Technical Reports Server (NTRS)

1992-01-01

An overview of the facilities and research being conducted at LeRC's Plumbrook field station is given. The video highlights four main structures and explains their uses. The Space Power Facility is the world's largest space environment simulation chamber, where spacebound hardware is tested in simulations of the vacuum and extreme heat and cold of the space plasma environment. This facility was used to prepare Atlas 1 rockets to ferry CRRES into orbit; it will also be used to test space nuclear electric power generation systems. The Spacecraft Propulsion Research Facility allows rocket vehicles to be hot fired in a simulated space environment. In the Cryogenic Propellant Tank Facility, researchers are developing technology for storing and transferring liquid hydrogen in space. There is also a Hypersonic Wind Tunnel which can perform flow tests with winds up to Mach 7.

Aerospace test facilities at NASA LERC Plumbrook

NASA Astrophysics Data System (ADS)

1992-10-01

An overview of the facilities and research being conducted at LeRC's Plumbrook field station is given. The video highlights four main structures and explains their uses. The Space Power Facility is the worlds largest space environment simulation chamber, where spacebound hardware is tested in simulations of the vacuum and extreme heat and cold of the space plasma environment. This facility was used to prepare Atlas 1 rockets to ferry CRRES into orbit; it will also be used to test space nuclear electric power generation systems. The Spacecraft Propulsion Research Facility allows rocket vehicles to be hot fired in a simulated space environment. In the Cryogenic Propellant Tank Facility, researchers are developing technology for storing and transferring liquid hydrogen in space. There is also a Hypersonic Wind Tunnel which can perform flow tests with winds up to Mach 7.

Heat induced temperature dysregulation and seizures in Dravet Syndrome/GEFS+ Gabrg2+/Q390X mice.

PubMed

Warner, Timothy A; Liu, Zhong; Macdonald, Robert L; Kang, Jing-Qiong

2017-08-01

It has been established that febrile seizures and its extended syndromes like generalized epilepsy with febrile seizures (FS) plus (GEFS+) and Dravet syndrome have been associated with mutations especially in SCN1A and GABRG2 genes. In patients, the onset of FS is likely due to the combined effect of temperature and inflammation in genetically vulnerable individuals because fever is often associated with infection. Much effort has been spent to understand the mechanisms underlying fever induction of seizures. In addition to the role of cytokines in FS, previous studies in Scn1a +/- knockout mice, a model of Dravet syndrome, indicated that temperature elevation alone could result in seizure generation, and the effect of elevated temperature inducing seizures was age-dependent. Here, we report the thermal effect in a different mouse model of Dravet syndrome, the Gabrg2 +/Q390X knockin mouse. We demonstrated age-dependent dysregulated temperature control and that temperature elevation produced myoclonic jerks, generalized tonic clonic seizures (GTCSs) and heightened anxiety-like symptoms in Gabrg2 +/Q390X mice. The study indicated that regardless of other inflammatory factors, brief heat alone increased brain excitability and induced multiple types of seizures in Gabrg2 +/Q390X mice, suggesting that mutations like GABRG2(Q390X) may alter brain thermal regulation and precipitate seizures during temperature elevations. Copyright © 2017 Elsevier B.V. All rights reserved.

Neutrophil-mediated oxidative burst and host defense are controlled by a Vav-PLCγ2 signaling axis in mice

PubMed Central

Graham, Daniel B.; Robertson, Charles M.; Bautista, Jhoanne; Mascarenhas, Francesca; Diacovo, M. Julia; Montgrain, Vivianne; Lam, Siu Kit; Cremasco, Viviana; Dunne, W. Michael; Faccio, Roberta; Coopersmith, Craig M.; Swat, Wojciech

2007-01-01

Oxidative burst, a critical antimicrobial mechanism of neutrophils, involves the rapid generation and release of reactive oxygen intermediates (ROIs) by the NADPH oxidase complex. Genetic mutations in an NADPH oxidase subunit, gp91 (also referred to as NOX2), are associated with chronic granulomatous disease (CGD), which is characterized by recurrent and life-threatening microbial infections. To combat such infections, ROIs are produced by neutrophils after stimulation by integrin-dependent adhesion to the ECM in conjunction with stimulation from inflammatory mediators, or microbial components containing pathogen-associated molecular patterns. In this report, we provide genetic evidence that both the Vav family of Rho GTPase guanine nucleotide exchange factors (GEFs) and phospholipase C–γ2 (PLC-γ2) are critical mediators of adhesion-dependent ROI production by neutrophils in mice. We also demonstrated that Vav was critically required for neutrophil-dependent host defense against systemic infection by Staphylococcus aureus and Pseudomonas aeruginosa, 2 common pathogens associated with fatal cases of hospital-acquired pneumonia. We identified a molecular pathway in which Vav GEFs linked integrin-mediated signaling with PLC-γ2 activation, release of intracellular Ca2+ cations, and generation of diacylglycerol to control assembly of the NADPH oxidase complex and ROI production by neutrophils. Taken together, our data indicate that integrin-dependent signals generated during neutrophil adhesion contribute to the activation of NADPH oxidase by a variety of distinct effector pathways, all of which require Vav. PMID:17932569

Theoretical and vibrational study of N-(3-chloro-4-fluoro-phenyl)-7-methoxy-6-(3-morpholin-4-ylpropoxy)-quinazolin-4-amine (gefitinib)

NASA Astrophysics Data System (ADS)

Mıhçıokur, Özlem; Özpozan, Talat

2015-12-01

N-(3-chloro-4fluoro-phenyl)-7-methoxy-6-(3-morpholin-4ylpropoxy)-quinazolin-4-amine (GEF), a quinalizoline derivative used as new anti-cancer agent, designed to target activity of epidermal growth factor receptor (EGFR) promoting the growth, division and spread of cancer cells, was examined from the vibrational and theoretical point of view. All calculations have been carried out both in gaseous and aqueous phases. In the calculations of both phases, the molecule has been optimized through conformer analysis beginning with the x-ray data. The conformer analyses have been carried out in each phases and the geometrical differences between the most stable structures in gaseous and in aqueous phases have been discussed. The solvent effect for GEF in aqueous solution was simulated by using self-consistent reaction field (SCRF) calculations employing the integral equation formalism variant (IEFPCM) model. NBO analysis has been performed to indicate the presence of intramolecular charge transfer. The complete assignments of the vibrational spectra (IR&Raman) were made with the aid of calculated spectra both in gaseous and aqueous phases. The observed spectral data of the title compound were compared with the calculated spectra obtained by DFT/B3LYP and DFT/B3PW91 methods using 6-31G(d,p) basis set. The theoretical results were found to be in good agreement with the measured experimental data especially for the interpretation of intra molecular interactions.

Stress- and Rho-activated ZO-1–associated nucleic acid binding protein binding to p21 mRNA mediates stabilization, translation, and cell survival

PubMed Central

Nie, Mei; Balda, Maria S.; Matter, Karl

2012-01-01

A central component of the cellular stress response is p21WAF1/CIP1, which regulates cell proliferation, survival, and differentiation. Inflammation and cell stress often up-regulate p21 posttranscriptionally by regulatory mechanisms that are poorly understood. ZO-1–associated nucleic acid binding protein (ZONAB)/DbpA is a Y-box transcription factor that is regulated by components of intercellular junctions that are affected by cytokines and tissue damage. We therefore asked whether ZONAB activation is part of the cellular stress response. Here, we demonstrate that ZONAB promotes cell survival in response to proinflammatory, hyperosmotic, and cytotoxic stress and that stress-induced ZONAB activation involves the Rho regulator GEF-H1. Unexpectedly, stress-induced ZONAB activation does not stimulate ZONAB’s activity as a transcription factor but leads to the posttranscriptional up-regulation of p21 protein and mRNA. Up-regulation is mediated by ZONAB binding to specific sites in the 3′-untranslated region of the p21 mRNA, resulting in mRNA stabilization and enhanced translation. Binding of ZONAB to mRNA is activated by GEF-H1 via Rho stimulation and also mediates Ras-induced p21 expression. We thus identify a unique type of stress and Rho signaling activated pathway that drives mRNA stabilization and translation and links the cellular stress response to p21 expression and cell survival. PMID:22711822

A new UK fission yield evaluation UKFY3.7

NASA Astrophysics Data System (ADS)

Mills, Robert William

2017-09-01

The JEFF neutron induced and spontaneous fission product yield evaluation is currently unchanged from JEFF-3.1.1, also known by its UK designation UKFY3.6A. It is based upon experimental data combined with empirically fitted mass, charge and isomeric state models which are then adjusted within the experimental and model uncertainties to conform to the physical constraints of the fission process. A new evaluation has been prepared for JEFF, called UKFY3.7, that incorporates new experimental data and replaces the current empirical models (multi-Gaussian fits of mass distribution and Wahl Zp model for charge distribution combined with parameter extrapolation), with predictions from GEF. The GEF model has the advantage that one set of parameters allows the prediction of many different fissioning nuclides at different excitation energies unlike previous models where each fissioning nuclide at a specific excitation energy had to be fitted individually to the relevant experimental data. The new UKFY3.7 evaluation, submitted for testing as part of JEFF-3.3, is described alongside initial results of testing. In addition, initial ideas for future developments allowing inclusion of new measurements types and changing from any neutron spectrum type to true neutron energy dependence are discussed. Also, a method is proposed to propagate uncertainties of fission product yields based upon the experimental data that underlies the fission yield evaluation. The covariance terms being determined from the evaluated cumulative and independent yields combined with the experimental uncertainties on the cumulative yield measurements.

The Lewis Research Center geomagnetic substorm simulation facility

NASA Technical Reports Server (NTRS)

Berkopec, F. D.; Stevens, N. J.; Sturman, J. C.

1977-01-01

A simulation facility was established to determine the response of typical spacecraft materials to the geomagnetic substorm environment and to evaluate instrumentation that will be used to monitor spacecraft system response to this environment. Space environment conditions simulated include the thermal-vacuum conditions of space, solar simulation, geomagnetic substorm electron fluxes and energies, and the low energy plasma environment. Measurements for spacecraft material tests include sample currents, sample surface potentials, and the cumulative number of discharges. Discharge transients are measured by means of current probes and oscilloscopes and are verified by a photomultiplier. Details of this facility and typical operating procedures are presented.

Wireless remote monitoring of critical facilities

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tsai, Hanchung; Anderson, John T.; Liu, Yung Y.

A method, apparatus, and system are provided for monitoring environment parameters of critical facilities. A Remote Area Modular Monitoring (RAMM) apparatus is provided for monitoring environment parameters of critical facilities. The RAMM apparatus includes a battery power supply and a central processor. The RAMM apparatus includes a plurality of sensors monitoring the associated environment parameters and at least one communication module for transmitting one or more monitored environment parameters. The RAMM apparatus is powered by the battery power supply, controlled by the central processor operating a wireless sensor network (WSN) platform when the facility condition is disrupted. The RAMM apparatusmore » includes a housing prepositioned at a strategic location, for example, where a dangerous build-up of contamination and radiation may preclude subsequent manned entrance and surveillance.« less

Accessibility of health clubs for people with mobility disabilities and visual impairments.

PubMed

Rimmer, James H; Riley, Barth; Wang, Edward; Rauworth, Amy

2005-11-01

We sought to examine the accessibility of health clubs to persons with mobility disabilities and visual impairments. We assessed 35 health clubs and fitness facilities as part of a national field trial of a new instrument, Accessibility Instruments Measuring Fitness and Recreation Environments (AIMFREE), designed to assess accessibility of fitness facilities in the following domains: (1) built environment, (2) equipment, (3) swimming pools, (4) information, (5) facility policies, and (6) professional behavior. All facilities had a low to moderate level of accessibility. Some of the deficiencies concerned specific Americans with Disabilities Act guidelines pertaining to the built environment, whereas other deficiency areas were related to aspects of the facilities' equipment, information, policies, and professional staff. Persons with mobility disabilities and visual impairments have difficulty accessing various areas of fitness facilities and health clubs. AIMFREE is an important tool for increasing awareness of these accessibility barriers for people with disabilities.

Two distinct mTORC2-dependent pathways converge on Rac1 to drive breast cancer metastasis.

PubMed

Morrison Joly, Meghan; Williams, Michelle M; Hicks, Donna J; Jones, Bayley; Sanchez, Violeta; Young, Christian D; Sarbassov, Dos D; Muller, William J; Brantley-Sieders, Dana; Cook, Rebecca S

2017-06-30

The importance of the mTOR complex 2 (mTORC2) signaling complex in tumor progression is becoming increasingly recognized. HER2-amplified breast cancers use Rictor/mTORC2 signaling to drive tumor formation, tumor cell survival and resistance to human epidermal growth factor receptor 2 (HER2)-targeted therapy. Cell motility, a key step in the metastatic process, can be activated by mTORC2 in luminal and triple negative breast cancer cell lines, but its role in promoting metastases from HER2-amplified breast cancers is not yet clear. Because Rictor is an obligate cofactor of mTORC2, we genetically engineered Rictor ablation or overexpression in mouse and human HER2-amplified breast cancer models for modulation of mTORC2 activity. Signaling through mTORC2-dependent pathways was also manipulated using pharmacological inhibitors of mTOR, Akt, and Rac. Signaling was assessed by western analysis and biochemical pull-down assays specific for Rac-GTP and for active Rac guanine nucleotide exchange factors (GEFs). Metastases were assessed from spontaneous tumors and from intravenously delivered tumor cells. Motility and invasion of cells was assessed using Matrigel-coated transwell assays. We found that Rictor ablation potently impaired, while Rictor overexpression increased, metastasis in spontaneous and intravenously seeded models of HER2-overexpressing breast cancers. Additionally, migration and invasion of HER2-amplified human breast cancer cells was diminished in the absence of Rictor, or upon pharmacological mTOR kinase inhibition. Active Rac1 was required for Rictor-dependent invasion and motility, which rescued invasion/motility in Rictor depleted cells. Rictor/mTORC2-dependent dampening of the endogenous Rac1 inhibitor RhoGDI2, a factor that correlated directly with increased overall survival in HER2-amplified breast cancer patients, promoted Rac1 activity and tumor cell invasion/migration. The mTORC2 substrate Akt did not affect RhoGDI2 dampening, but partially increased Rac1 activity through the Rac-GEF Tiam1, thus partially rescuing cell invasion/motility. The mTORC2 effector protein kinase C (PKC)α did rescue Rictor-mediated RhoGDI2 downregulation, partially rescuing Rac-guanosine triphosphate (GTP) and migration/motility. These findings suggest that mTORC2 uses two coordinated pathways to activate cell invasion/motility, both of which converge on Rac1. Akt signaling activates Rac1 through the Rac-GEF Tiam1, while PKC signaling dampens expression of the endogenous Rac1 inhibitor, RhoGDI2.

The Acoustic Environment of the NASA Glenn 9- by 15-foot Low-Speed Wind Tunnel

NASA Technical Reports Server (NTRS)

Stephens, David B.

2015-01-01

The 9- by 15-Foot Low Speed Wind Tunnel is an acoustic testing facility with a long history of aircraft propulsion noise research. Due to interest in renovating the facility to support future testing of advanced quiet engine designs, a study was conducted to document the background noise level in the facility and investigate the sources of contaminating noise. The anechoic quality of the facility was also investigated using an interrupted noise method. The present report discusses these aspects of the noise environment in this facility.

Data Acquisition System Architecture and Capabilities At NASA GRC Plum Brook Station's Space Environment Test Facilities

NASA Technical Reports Server (NTRS)

Evans, Richard K.; Hill, Gerald M.

2012-01-01

Very large space environment test facilities present unique engineering challenges in the design of facility data systems. Data systems of this scale must be versatile enough to meet the wide range of data acquisition and measurement requirements from a diverse set of customers and test programs, but also must minimize design changes to maintain reliability and serviceability. This paper presents an overview of the common architecture and capabilities of the facility data acquisition systems available at two of the world?s largest space environment test facilities located at the NASA Glenn Research Center?s Plum Brook Station in Sandusky, Ohio; namely, the Space Propulsion Research Facility (commonly known as the B-2 facility) and the Space Power Facility (SPF). The common architecture of the data systems is presented along with details on system scalability and efficient measurement systems analysis and verification. The architecture highlights a modular design, which utilizes fully-remotely managed components, enabling the data systems to be highly configurable and support multiple test locations with a wide-range of measurement types and very large system channel counts.

Data Acquisition System Architecture and Capabilities at NASA GRC Plum Brook Station's Space Environment Test Facilities

NASA Technical Reports Server (NTRS)

Evans, Richard K.; Hill, Gerald M.

2014-01-01

Very large space environment test facilities present unique engineering challenges in the design of facility data systems. Data systems of this scale must be versatile enough to meet the wide range of data acquisition and measurement requirements from a diverse set of customers and test programs, but also must minimize design changes to maintain reliability and serviceability. This paper presents an overview of the common architecture and capabilities of the facility data acquisition systems available at two of the world's largest space environment test facilities located at the NASA Glenn Research Center's Plum Brook Station in Sandusky, Ohio; namely, the Space Propulsion Research Facility (commonly known as the B-2 facility) and the Space Power Facility (SPF). The common architecture of the data systems is presented along with details on system scalability and efficient measurement systems analysis and verification. The architecture highlights a modular design, which utilizes fully-remotely managed components, enabling the data systems to be highly configurable and support multiple test locations with a wide-range of measurement types and very large system channel counts.

40 CFR Appendix G to Part 112 - Tier I Qualified Facility SPCC Plan

Code of Federal Regulations, 2013 CFR

2013-07-01

... 40 Protection of Environment 23 2013-07-01 2013-07-01 false Tier I Qualified Facility SPCC Plan G Appendix G to Part 112 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS OIL POLLUTION PREVENTION Pt. 112, App. G Appendix G to Part 112—Tier I Qualified Facility SPCC Plan...

40 CFR Appendix G to Part 112 - Tier I Qualified Facility SPCC Plan

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 23 2012-07-01 2012-07-01 false Tier I Qualified Facility SPCC Plan G Appendix G to Part 112 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS OIL POLLUTION PREVENTION Pt. 112, App. G Appendix G to Part 112—Tier I Qualified Facility SPCC Plan...

40 CFR Appendix G to Part 112 - Tier I Qualified Facility SPCC Plan

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 21 2010-07-01 2010-07-01 false Tier I Qualified Facility SPCC Plan G Appendix G to Part 112 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS OIL POLLUTION PREVENTION Pt. 112, App. G Appendix G to Part 112—Tier I Qualified Facility SPCC Plan...

40 CFR Appendix G to Part 112 - Tier I Qualified Facility SPCC Plan

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 22 2011-07-01 2011-07-01 false Tier I Qualified Facility SPCC Plan G Appendix G to Part 112 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS OIL POLLUTION PREVENTION Pt. 112, App. G Appendix G to Part 112—Tier I Qualified Facility SPCC Plan...

40 CFR Appendix G to Part 112 - Tier I Qualified Facility SPCC Plan

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 22 2014-07-01 2013-07-01 true Tier I Qualified Facility SPCC Plan G Appendix G to Part 112 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS OIL POLLUTION PREVENTION Pt. 112, App. G Appendix G to Part 112—Tier I Qualified Facility SPCC Plan...

76 FR 37799 - DOE Final Decision in Response to Recommendation 2010-1 of the Defense Nuclear Facilities Safety...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-06-28

... the public, workers, and the environment. For example, the Board clarified that use of the term.... Department of Energy Nonreactor Nuclear Facility Documented Safety Analyses, as a safe harbor methodology..., our workers, and the environment at all of our facilities. We share your conviction that a clear set...

STS-60 Cosmonauts in Weightless Environment Training Facility (WETF) training

NASA Image and Video Library

1993-01-07

Russian Cosmonaut Vladimir Titov maneuvers a small life raft during bailout training at JSC's Weightless Environment Training Facility (WETF). Two SCUBA-equipped divers assisted Titov in the STS-60 training exercise.

Netz ohne doppelten Boden: Internet-Sicherheit

NASA Astrophysics Data System (ADS)

Fichtner, Johann

2002-07-01

Die Sicherheitslücken des heutigen Internets haben zwei Ursachen: sein ursprüngliches Design für rein militärische Zwecke sowie die gefährliche Sorglosigkeit der Entwickler und Betreiber von Internet-Komponenten. Dabei kann ein kleiner Kreis von Experten Angriffe auf die eigentlichen Netzknoten ausführen. Typische Hacker-Angriffe kommen von Endsystemen des Internets und richten sich auch gegen Endsysteme wie Firmen-Intranets oder einzelne PCs. Dagegen gibt es eine Reihe von Schutzmöglichkeiten. Es ist jedoch keine einfache Lösung zur Erhöhung der Internet-Sicherheit in Sicht.

Emerging CAE technologies and their role in Future Ambient Intelligence Environments

NASA Astrophysics Data System (ADS)

Noor, Ahmed K.

2011-03-01

Dramatic improvements are on the horizon in Computer Aided Engineering (CAE) and various simulation technologies. The improvements are due, in part, to the developments in a number of leading-edge technologies and their synergistic combinations/convergence. The technologies include ubiquitous, cloud, and petascale computing; ultra high-bandwidth networks, pervasive wireless communication; knowledge based engineering; networked immersive virtual environments and virtual worlds; novel human-computer interfaces; and powerful game engines and facilities. This paper describes the frontiers and emerging simulation technologies, and their role in the future virtual product creation and learning/training environments. The environments will be ambient intelligence environments, incorporating a synergistic combination of novel agent-supported visual simulations (with cognitive learning and understanding abilities); immersive 3D virtual world facilities; development chain management systems and facilities (incorporating a synergistic combination of intelligent engineering and management tools); nontraditional methods; intelligent, multimodal and human-like interfaces; and mobile wireless devices. The Virtual product creation environment will significantly enhance the productivity and will stimulate creativity and innovation in future global virtual collaborative enterprises. The facilities in the learning/training environment will provide timely, engaging, personalized/collaborative and tailored visual learning.

Microgravity science and applications: Apparatus and facilities

NASA Technical Reports Server (NTRS)

1989-01-01

NASA support apparatus and facilities for microgravity research are summarized in fact sheets. The facilities are ground-based simulation environments for short-term experiments, and the shuttle orbiter environment for long duration experiments. The 17 items of the microgravitational experimental apparatus are described. Electronic materials, alloys, biotechnology, fluid dynamics and transport phenomena, glasses and ceramics, and combustion science are among the topics covered.

A radiant heating test facility for space shuttle orbiter thermal protection system certification

NASA Technical Reports Server (NTRS)

Sherborne, W. D.; Milhoan, J. D.

1980-01-01

A large scale radiant heating test facility was constructed so that thermal certification tests can be performed on the new generation of thermal protection systems developed for the space shuttle orbiter. This facility simulates surface thermal gradients, onorbit cold-soak temperatures down to 200 K, entry heating temperatures to 1710 K in an oxidizing environment, and the dynamic entry pressure environment. The capabilities of the facility and the development of new test equipment are presented.

A Phenomenological Study of the Work Environment in Long-Term Care Facilities for the Older Adults.

PubMed

Choi, Sandy Pin Pin; Yeung, Cheryl Chi Yan; Lee, Joseph Kok Long

2018-05-01

Attempts to meet the increasing demand for long-term care (LTC) services have been hindered by acute staff shortages and high turnover. Distinct from previous studies, a descriptive phenomenological approach with van Kaam's controlled explication method was adopted in this study, to delineate how attributes of the LTC work environment shape the workforce crisis. Individual interviews were conducted with 40 LTC workers from 10 facilities in Hong Kong. The results suggest that the work environment in LTC facilities is not only characterized by organization- and job-related attributes that influence staff outcomes but also is a socially constructed concept with derogatory connotations that can influence staff recruitment and retention. Concerted efforts from facility administrators and policy makers are needed to improve the quality of the work environment. Future initiatives should focus on developing a vision and strategic plan to facilitate the rise of the LTC sector as a profession.

Adopting and implementing nutrition guidelines in recreational facilities: Public and private sector roles. A multiple case study

PubMed Central

2012-01-01

Background Recreational facilities are an important community resource for health promotion because they provide access to affordable physical activities. However, despite their health mandate, many have unhealthy food environments that may paradoxically increase the risk of childhood obesity. The Alberta Nutrition Guidelines for Children and Youth (ANGCY) are government-initiated, voluntary guidelines intended to facilitate children’s access to healthy food and beverage choices in schools, childcare and recreational facilities, however few recreational facilities are using them. Methods We used mixed methods within an exploratory multiple case study to examine factors that influenced adoption and implementation of the ANGCY and the nature of the food environment within three cases: an adopter, a semi-adopter and a non-adopter of the ANGCY. Diffusion of Innovations theory provided the theoretical platform for the study. Qualitative data were generated through interviews, observations, and document reviews, and were analysed using directed content analysis. Set theoretic logic was used to identify factors that differentiated adopters from the non-adopter. Quantitative sales data were also collected, and the quality of the food environment was scored using four complementary tools. Results The keys to adoption and implementation of nutrition guidelines in recreational facilities related to the managers’ nutrition-related knowledge, beliefs and perceptions, as these shaped his decisions and actions. The manager, however, could not accomplish adoption and implementation alone. Intersectoral linkages with schools and formal, health promoting partnerships with industry were also important for adoption and implementation to occur. The food environment in facilities that had adopted the ANGCY did not appear to be superior to the food environment in facilities that had not adopted the ANGCY. Conclusions ANGCY uptake may continue to falter under the current voluntary approach, as the environmental supports for voluntary action are poor. Where ANGCY uptake does occur, changes to the food environment may be relatively minor. Stronger government measures may be needed to require recreational facilities to improve their food environments and to limit availability of unhealthy foods. PMID:22632384

Publically funded recreation facilities: obesogenic environments for children and families?

PubMed

Naylor, Patti-Jean; Bridgewater, Laura; Purcell, Megan; Ostry, Aleck; Wekken, Suzanne Vander

2010-05-01

Increasing healthy food options in public venues, including recreational facilities, is a health priority. The purpose of this study was to describe the public recreation food environment in British Columbia, Canada using a sequential explanatory mixed methods design. Facility audits assessed policy, programs, vending, concessions, fundraising, staff meetings and events. Focus groups addressed context and issues related to action. Eighty-eighty percent of facilities had no policy governing food sold or provided for children/youth programs. Sixty-eight percent of vending snacks were chocolate bars and chips while 57% of beverages were sugar sweetened. User group fundraisers held at the recreation facilities also sold 'unhealthy' foods. Forty-two percent of recreation facilities reported providing user-pay programs that educated the public about healthy eating. Contracts, economics, lack of resources and knowledge and motivation of staff and patrons were barriers to change. Recreation food environments were obesogenic but stakeholders were interested in change. Technical support, resources and education are needed.

Publically Funded Recreation Facilities: Obesogenic Environments for Children and Families?

PubMed Central

Naylor, Patti-Jean; Bridgewater, Laura; Purcell, Megan; Ostry, Aleck; Wekken, Suzanne Vander

2010-01-01

Increasing healthy food options in public venues, including recreational facilities, is a health priority. The purpose of this study was to describe the public recreation food environment in British Columbia, Canada using a sequential explanatory mixed methods design. Facility audits assessed policy, programs, vending, concessions, fundraising, staff meetings and events. Focus groups addressed context and issues related to action. Eighty-eighty percent of facilities had no policy governing food sold or provided for children/youth programs. Sixty-eight percent of vending snacks were chocolate bars and chips while 57% of beverages were sugar sweetened. User group fundraisers held at the recreation facilities also sold ‘unhealthy’ foods. Forty-two percent of recreation facilities reported providing user-pay programs that educated the public about healthy eating. Contracts, economics, lack of resources and knowledge and motivation of staff and patrons were barriers to change. Recreation food environments were obesogenic but stakeholders were interested in change. Technical support, resources and education are needed. PMID:20623020

78 FR 76344 - Self-Regulatory Organizations; NASDAQ OMX BX, Inc.; Notice of Filing and Immediate Effectiveness...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-12-17

... environment and on which they may test their automated systems that integrate with the Exchange. For example... Establish Fees Under Rule 7030(d) for Use of the Carteret Testing Facility Test Environment December 11... the Testing Facility (``NTF'') test environment located in Carteret, New Jersey, which will provide a...

78 FR 76350 - Self-Regulatory Organizations; NASDAQ OMX PHLX LLC.; Notice of Filing and Immediate Effectiveness...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-12-17

... switch port in the test environment. The test environment is designed to closely mirror the live equity... Change To Establish Fees Under the PHLX Pricing Schedule for Use of the Carteret Testing Facility Test... establish fees under the Pricing Schedule for use of the Testing Facility (``NTF'') test environment located...

33 CFR 154.1240 - Specific requirements for animal fats and vegetable oils facilities that could reasonably be...

Code of Federal Regulations, 2012 CFR

2012-07-01

... environment. 154.1240 Section 154.1240 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND... oils facilities that could reasonably be expected to cause substantial harm to the environment. (a) The... expected to cause substantial harm to the environment, must submit a response plan that meets the...

33 CFR 154.1240 - Specific requirements for animal fats and vegetable oils facilities that could reasonably be...

Code of Federal Regulations, 2014 CFR

2014-07-01

... environment. 154.1240 Section 154.1240 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND... oils facilities that could reasonably be expected to cause substantial harm to the environment. (a) The... expected to cause substantial harm to the environment, must submit a response plan that meets the...

33 CFR 154.1240 - Specific requirements for animal fats and vegetable oils facilities that could reasonably be...

Code of Federal Regulations, 2013 CFR

2013-07-01

... environment. 154.1240 Section 154.1240 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND... oils facilities that could reasonably be expected to cause substantial harm to the environment. (a) The... expected to cause substantial harm to the environment, must submit a response plan that meets the...

33 CFR 154.1240 - Specific requirements for animal fats and vegetable oils facilities that could reasonably be...

Code of Federal Regulations, 2010 CFR

2010-07-01

... environment. 154.1240 Section 154.1240 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND... oils facilities that could reasonably be expected to cause substantial harm to the environment. (a) The... expected to cause substantial harm to the environment, must submit a response plan that meets the...

Fusion interfaces for tactical environments: An application of virtual reality technology

NASA Technical Reports Server (NTRS)

Haas, Michael W.

1994-01-01

The term Fusion Interface is defined as a class of interface which integrally incorporates both virtual and nonvirtual concepts and devices across the visual, auditory, and haptic sensory modalities. A fusion interface is a multisensory virtually-augmented synthetic environment. A new facility has been developed within the Human Engineering Division of the Armstrong Laboratory dedicated to exploratory development of fusion interface concepts. This new facility, the Fusion Interfaces for Tactical Environments (FITE) Facility is a specialized flight simulator enabling efficient concept development through rapid prototyping and direct experience of new fusion concepts. The FITE Facility also supports evaluation of fusion concepts by operation fighter pilots in an air combat environment. The facility is utilized by a multidisciplinary design team composed of human factors engineers, electronics engineers, computer scientists, experimental psychologists, and oeprational pilots. The FITE computational architecture is composed of twenty-five 80486-based microcomputers operating in real-time. The microcomputers generate out-the-window visuals, in-cockpit and head-mounted visuals, localized auditory presentations, haptic displays on the stick and rudder pedals, as well as executing weapons models, aerodynamic models, and threat models.

Postirradiation Testing Laboratory (327 Building)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kammenzind, D.E.

A Standards/Requirements Identification Document (S/RID) is the total list of the Environment, Safety and Health (ES and H) requirements to be implemented by a site, facility, or activity. These requirements are appropriate to the life cycle phase to achieve an adequate level of protection for worker and public health and safety, and the environment during design, construction, operation, decontamination and decommissioning, and environmental restoration. S/RlDs are living documents, to be revised appropriately based on change in the site`s or facility`s mission or configuration, a change in the facility`s life cycle phase, or a change to the applicable standards/requirements. S/RIDs encompassmore » health and safety, environmental, and safety related safeguards and security (S and S) standards/requirements related to the functional areas listed in the US Department of Energy (DOE) Environment, Safety and Health Configuration Guide. The Fluor Daniel Hanford (FDH) Contract S/RID contains standards/requirements, applicable to FDH and FDH subcontractors, necessary for safe operation of Project Hanford Management Contract (PHMC) facilities, that are not the direct responsibility of the facility manager (e.g., a site-wide fire department). Facility S/RIDs contain standards/requirements applicable to a specific facility that are the direct responsibility of the facility manager. S/RlDs are prepared by those responsible for managing the operation of facilities or the conduct of activities that present a potential threat to the health and safety of workers, public, or the environment, including: Hazard Category 1 and 2 nuclear facilities and activities, as defined in DOE 5480.23. Selected Hazard Category 3 nuclear, and Low Hazard non-nuclear facilities and activities, as agreed upon by RL. The Postirradiation Testing Laboratory (PTL) S/RID contains standards/ requirements that are necessary for safe operation of the PTL facility, and other building/areas that are the direct responsibility of the specific facility manager. The specific DOE Orders, regulations, industry codes/standards, guidance documents and good industry practices that serve as the basis for each element/subelement are identified and aligned with each subelement.« less

40 CFR 256.25 - Recommendation for inactive facilities.

Code of Federal Regulations, 2014 CFR

2014-07-01

... notification of agencies responsible for public health and safety. ... 40 Protection of Environment 25 2014-07-01 2014-07-01 false Recommendation for inactive facilities. 256.25 Section 256.25 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID...

40 CFR 256.25 - Recommendation for inactive facilities.

Code of Federal Regulations, 2012 CFR

2012-07-01

... notification of agencies responsible for public health and safety. ... 40 Protection of Environment 26 2012-07-01 2011-07-01 true Recommendation for inactive facilities. 256.25 Section 256.25 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID...

40 CFR 256.25 - Recommendation for inactive facilities.

Code of Federal Regulations, 2013 CFR

2013-07-01

... notification of agencies responsible for public health and safety. ... 40 Protection of Environment 26 2013-07-01 2013-07-01 false Recommendation for inactive facilities. 256.25 Section 256.25 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID...

Accessibility of Health Clubs for People with Mobility Disabilities and Visual Impairments

PubMed Central

Rimmer, James H.; Riley, Barth; Wang, Edward; Rauworth, Amy

2005-01-01

Objective. We sought to examine the accessibility of health clubs to persons with mobility disabilities and visual impairments. Methods. We assessed 35 health clubs and fitness facilities as part of a national field trial of a new instrument, Accessibility Instruments Measuring Fitness and Recreation Environments (AIMFREE), designed to assess accessibility of fitness facilities in the following domains: (1) built environment, (2) equipment, (3) swimming pools, (4) information, (5) facility policies, and (6) professional behavior. Results. All facilities had a low to moderate level of accessibility. Some of the deficiencies concerned specific Americans with Disabilities Act guidelines pertaining to the built environment, whereas other deficiency areas were related to aspects of the facilities’ equipment, information, policies, and professional staff. Conclusions. Persons with mobility disabilities and visual impairments have difficulty accessing various areas of fitness facilities and health clubs. AIMFREE is an important tool for increasing awareness of these accessibility barriers for people with disabilities. PMID:16254234

The Chromobacterium violaceum type III effector CopE, a guanine nucleotide exchange factor for Rac1 and Cdc42, is involved in bacterial invasion of epithelial cells and pathogenesis.

PubMed

Miki, Tsuyoshi; Akiba, Kinari; Iguchi, Mirei; Danbara, Hirofumi; Okada, Nobuhiko

2011-06-01

The type III secretion system (T3SS) encoded by Chromobacterium pathogenicity islands 1 and 1a (Cpi-1/-1a) is critical for Chromobacterium violaceum pathogenesis. T3SS-dependent virulence is commonly characterized by type III effector virulence function, but the full repertoire of the effector proteins of Cpi-1/-1a T3SS is unknown. In this study, we showed that expression of Cpi-1/-1a T3SS is controlled by the master regulator CilA. We used transcriptional profiling with DNA microarrays to define CilA regulon and identified genes encoding T3SS effectors whose translocation into host cells was dependent on Cpi-1/-1a T3SS. From these effectors, we found that CopE (CV0296) has similarities to a guanine nucleotide exchange factor (GEF) for Rho GTPases in its C-terminal portion. The N-terminal portions (1-81 amino acids) of CopE and a CivB as a putative chaperone were required for its translocation. CopE specifically activates Rac1 and Cdc42 followed by the induction of actin cytoskeletal rearrangement. Interestingly, C. violaceum invades human epithelial HeLa cells in a Cpi-1/-1a-encoded T3SS- and CopE-dependent manner. Finally, C. violaceum strains lacking copE and expressing a CopE-G168V deficient in GEF activity were attenuated for virulence in mice, suggesting that CopE contributes to the virulence of this pathogen. © 2011 Blackwell Publishing Ltd.

The Guanine Nucleotide Exchange Factor Tiam1 Affects Neuronal Morphology; Opposing Roles for the Small GTPases Rac and Rho

PubMed Central

van Leeuwen, Frank N.; Kain, Hendrie E.T.; van der Kammen, Rob A.; Michiels, Frits; Kranenburg, Onno W.; Collard, John G.

1997-01-01

The invasion-inducing T-lymphoma invasion and metastasis 1 (Tiam1) protein functions as a guanine nucleotide exchange factor (GEF) for the small GTPase Rac1. Differentiation-dependent expression of Tiam1 in the developing brain suggests a role for this GEF and its effector Rac1 in the control of neuronal morphology. Here we show that overexpression of Tiam1 induces cell spreading and affects neurite outgrowth in N1E-115 neuroblastoma cells. These effects are Rac-dependent and strongly promoted by laminin. Overexpression of Tiam1 recruits the α6β1 integrin, a laminin receptor, to specific adhesive contacts at the cell periphery, which are different from focal contacts. Cells overexpressing Tiam1 no longer respond to lysophosphatidic acid– induced neurite retraction and cell rounding, processes mediated by Rho, suggesting that Tiam1-induced activation of Rac antagonizes Rho signaling. This inhibition can be overcome by coexpression of constitutively active RhoA, which may indicate that regulation occurs at the level of Rho or upstream. Conversely, neurite formation induced by Tiam1 or Rac1 is further promoted by inactivating Rho. These results demonstrate that Rac- and Rho-mediated pathways oppose each other during neurite formation and that a balance between these pathways determines neuronal morphology. Furthermore, our data underscore the potential role of Tiam1 as a specific regulator of Rac during neurite formation and illustrate the importance of reciprocal interactions between the cytoskeleton and the extracellular matrix during this process. PMID:9348295

A structural study of the complex between neuroepithelial cell transforming gene 1 (Net1) and RhoA reveals a potential anticancer drug hot spot.

PubMed

Petit, Alain-Pierre; Garcia-Petit, Christel; Bueren-Calabuig, Juan A; Vuillard, Laurent M; Ferry, Gilles; Boutin, Jean A

2018-06-08

The GTPase RhoA is a major player in many different regulatory pathways. RhoA catalyzes GTP hydrolysis, and its catalysis is accelerated when RhoA forms heterodimers with proteins of the guanine nucleotide exchange factor (GEF) family. Neuroepithelial cell transforming gene 1 (Net1) is a RhoA-interacting GEF implicated in cancer, but the structural features supporting the RhoA/Net1 interaction are unknown. Taking advantage of a simple production and purification process, here we solved the structure of a RhoA/Net1 heterodimer with X-ray crystallography at 2-Å resolution. Using a panel of several techniques, including molecular dynamics simulations, we characterized the RhoA/Net1 interface. Moreover, deploying an extremely simple peptide-based scanning approach, we found that short peptides (penta- to nonapeptides) derived from the protein/protein interaction region of RhoA could disrupt the RhoA/Net1 interaction and thereby diminish the rate of nucleotide exchange. The most inhibitory peptide, EVKHF, spanning residues 102-106 in the RhoA sequence, displayed an IC 50 of ∼100 μm without further modifications. The peptides identified here could be useful in further investigations of the RhoA/Net1 interaction region. We propose that our structural and functional insights might inform chemical approaches for transforming the pentapeptide into an optimized pseudopeptide that antagonizes Net1-mediated RhoA activation with therapeutic anticancer potential. © 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

A critical role for phosphatidylinositol (3,4,5)-trisphosphate-dependent Rac exchanger 1 in endothelial junction disruption and vascular hyperpermeability

PubMed Central

Naikawadi, Ram P.; Cheng, Ni; Vogel, Stephen M.; Qian, Feng; Wu, Dianqing; Malik, Asrar B.; Ye, Richard D.

2013-01-01

Rationale The small GTPase Rac is critical to vascular endothelial functions, yet its regulation in endothelial cells remains unclear. Understanding the upstream pathway may delineate Rac activation mechanisms and its role in maintaining vascular endothelial barrier integrity. Objective By investigating P-Rex1, one of the Rac-specific guanine nucleotide exchange factors (GEFs) previously known for G protein-coupled receptor (GPCR) signaling, we sought to determine whether Rac-GEF is a nodal for signal integration and potential target for drug intervention. Methods and Results Using gene deletion and siRNA silencing approach, we investigated the role of P-Rex1 in lung microvascular endothelial cells (HLMVECs). TNF-α exposure led to disruption of endothelial junctions, and silencing P-Rex1 protected junction integrity. TNF-α stimulated Rac activation and ROS production in a P-Rex1-dependent manner. Removal of P-Rex1 significantly reduced ICAM-1 expression, PMN transendothelial migration and leukocyte sequestration in TNF-α challenged mouse lungs. The P-Rex1 knockout mice were also refractory to lung vascular hyper-permeability and edema in a LPS-induced sepsis model. Conclusions These results demonstrate for the first time that P-Rex1 expressed in endothelial cells is activated downstream of TNF-α, which is not a GPCR agonist. Our data identify P-Rex1 as a critical mediator of vascular barrier disruption. Targeting P-Rex1 may effectively protect against TNF-α and LPS-induced endothelial junction disruption and vascular hyper-permeability. PMID:22965143

Functional Analysis of BcBem1 and Its Interaction Partners in Botrytis cinerea: Impact on Differentiation and Virulence

PubMed Central

Schumacher, Julia; Kokkelink, Leonie; Tudzynski, Paul

2014-01-01

In phytopathogenic fungi the establishment and maintenance of polarity is not only essential for vegetative growth and differentiation, but also for penetration and colonization of host tissues. We investigated orthologs of members of the yeast polarity complex in the grey mould fungus Botrytis cinerea: the scaffold proteins Bem1 and Far1, the GEF (guanine nucleotide exchange factor) Cdc24, and the formin Bni1 (named Sep1 in B. cinerea). BcBem1 does not play an important role in regular hyphal growth, but has significant impact on spore formation and germination, on the establishment of conidial anastomosis tubes (CATs) and on virulence. As in other fungi, BcBem1 interacts with the GEF BcCdc24 and the formin BcSep1, indicating that in B. cinerea the apical complex has a similar structure as in yeast. A functional analysis of BcCdc24 suggests that it is essential for growth, since it was not possible to obtain homokaryotic deletion mutants. Heterokaryons of Δcdc24 (supposed to exhibit reduced bccdc24 transcript levels) already show a strong phenotype: an inability to penetrate the host tissue, a significantly reduced growth rate and malformation of conidia, which tend to burst as observed for Δbcbem1. Also the formin BcSep1 has significant impact on hyphal growth and development, whereas the role of the putative ortholog of the yeast scaffold protein Far1 remains open: Δbcfar1 mutants have no obvious phenotypes. PMID:24797931

Activation of Extracellular Signal-Regulated Kinase but Not of p38 Mitogen-Activated Protein Kinase Pathways in Lymphocytes Requires Allosteric Activation of SOS

PubMed Central

Jun, Jesse E.; Yang, Ming; Chen, Hang; Chakraborty, Arup K.

2013-01-01

Thymocytes convert graded T cell receptor (TCR) signals into positive selection or deletion, and activation of extracellular signal-related kinase (ERK), p38, and Jun N-terminal protein kinase (JNK) mitogen-activated protein kinases (MAPKs) has been postulated to play a discriminatory role. Two families of Ras guanine nucleotide exchange factors (RasGEFs), SOS and RasGRP, activate Ras and the downstream RAF-MEK-ERK pathway. The pathways leading to lymphocyte p38 and JNK activation are less well defined. We previously described how RasGRP alone induces analog Ras-ERK activation while SOS and RasGRP cooperate to establish bimodal ERK activation. Here we employed computational modeling and biochemical experiments with model cell lines and thymocytes to show that TCR-induced ERK activation grows exponentially in thymocytes and that a W729E allosteric pocket mutant, SOS1, can only reconstitute analog ERK signaling. In agreement with RasGRP allosterically priming SOS, exponential ERK activation is severely decreased by pharmacological or genetic perturbation of the phospholipase Cγ (PLCγ)-diacylglycerol-RasGRP1 pathway. In contrast, p38 activation is not sharply thresholded and requires high-level TCR signal input. Rac and p38 activation depends on SOS1 expression but not allosteric activation. Based on computational predictions and experiments exploring whether SOS functions as a RacGEF or adaptor in Rac-p38 activation, we established that the presence of SOS1, but not its enzymatic activity, is critical for p38 activation. PMID:23589333

Structural basis for activation of trimeric Gi proteins by multiple growth factor receptors via GIV/Girdin

PubMed Central

Lin, Changsheng; Ear, Jason; Midde, Krishna; Lopez-Sanchez, Inmaculada; Aznar, Nicolas; Garcia-Marcos, Mikel; Kufareva, Irina; Abagyan, Ruben; Ghosh, Pradipta

2014-01-01

A long-standing issue in the field of signal transduction is to understand the cross-talk between receptor tyrosine kinases (RTKs) and heterotrimeric G proteins, two major and distinct signaling hubs that control eukaryotic cell behavior. Although stimulation of many RTKs leads to activation of trimeric G proteins, the molecular mechanisms behind this phenomenon remain elusive. We discovered a unifying mechanism that allows GIV/Girdin, a bona fide metastasis-related protein and a guanine-nucleotide exchange factor (GEF) for Gαi, to serve as a direct platform for multiple RTKs to activate Gαi proteins. Using a combination of homology modeling, protein–protein interaction, and kinase assays, we demonstrate that a stretch of ∼110 amino acids within GIV C-terminus displays structural plasticity that allows folding into a SH2-like domain in the presence of phosphotyrosine ligands. Using protein–protein interaction assays, we demonstrated that both SH2 and GEF domains of GIV are required for the formation of a ligand-activated ternary complex between GIV, Gαi, and growth factor receptors and for activation of Gαi after growth factor stimulation. Expression of a SH2-deficient GIV mutant (Arg 1745→Leu) that cannot bind RTKs impaired all previously demonstrated functions of GIV—Akt enhancement, actin remodeling, and cell migration. The mechanistic and structural insights gained here shed light on the long-standing questions surrounding RTK/G protein cross-talk, set a novel paradigm, and characterize a unique pharmacological target for uncoupling GIV-dependent signaling downstream of multiple oncogenic RTKs. PMID:25187647

28 CFR Appendix C to Part 61 - Immigration and Naturalization Service Procedures Relating to the Implementation of the National...

Code of Federal Regulations, 2011 CFR

2011-07-01

... INS facility which would have a significant impact upon the environment. (b) Construction of a new... have a significant impact upon the environment. (Reference: § 1507.3(b)(2)(i)—CEQ Regulations.) 10... significantly impact upon the environment. (b) Increase or decrease in population of a facility within its...

40 CFR 61.20 - Designation of facilities.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 8 2010-07-01 2010-07-01 false Designation of facilities. 61.20 Section 61.20 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) NATIONAL EMISSION STANDARDS FOR HAZARDOUS AIR POLLUTANTS National Emission Standards for Radon...

40 CFR 61.250 - Designation of facilities.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 8 2010-07-01 2010-07-01 false Designation of facilities. 61.250 Section 61.250 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) NATIONAL EMISSION STANDARDS FOR HAZARDOUS AIR POLLUTANTS National Emission Standards for Radon...

40 CFR 61.220 - Designation of facilities.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 8 2010-07-01 2010-07-01 false Designation of facilities. 61.220 Section 61.220 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) NATIONAL EMISSION STANDARDS FOR HAZARDOUS AIR POLLUTANTS National Emission Standards for Radon...

40 CFR 61.200 - Designation of facilities.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 8 2010-07-01 2010-07-01 false Designation of facilities. 61.200 Section 61.200 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) NATIONAL EMISSION STANDARDS FOR HAZARDOUS AIR POLLUTANTS National Emission Standards for Radon...

An effective combined environment test facility

NASA Technical Reports Server (NTRS)

Deitch, A.

1980-01-01

A critical missile component required operational verification while subjected to combined environments within and beyond flight parameters. The testing schedule necessitated the design and fabrication of a test facility in order to provide the specified temperatures combined with humidity, altitude and vibration.

40 CFR 61.250 - Designation of facilities.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 8 2011-07-01 2011-07-01 false Designation of facilities. 61.250 Section 61.250 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) NATIONAL EMISSION STANDARDS FOR HAZARDOUS AIR POLLUTANTS National Emission Standards for Radon...

40 CFR 61.20 - Designation of facilities.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 8 2011-07-01 2011-07-01 false Designation of facilities. 61.20 Section 61.20 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) NATIONAL EMISSION STANDARDS FOR HAZARDOUS AIR POLLUTANTS National Emission Standards for Radon...

40 CFR 61.200 - Designation of facilities.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 8 2011-07-01 2011-07-01 false Designation of facilities. 61.200 Section 61.200 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) NATIONAL EMISSION STANDARDS FOR HAZARDOUS AIR POLLUTANTS National Emission Standards for Radon...

40 CFR 61.220 - Designation of facilities.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 8 2011-07-01 2011-07-01 false Designation of facilities. 61.220 Section 61.220 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) NATIONAL EMISSION STANDARDS FOR HAZARDOUS AIR POLLUTANTS National Emission Standards for Radon...

Facility Management's Role in Organizational Sustainability

ERIC Educational Resources Information Center

Adams, Gregory K.

2013-01-01

Facility managers have questions about sustainability. How do an organization's physical facilities--its built environment--and the management of them, influence the sustainability of the organization or institution as a whole? How important is Facility Management (FM) to the overall sustainability profile of an organization? Facility managers…

Small Projects Rapid Integration and Test Environment (SPRITE): Application for Increasing Robustness

NASA Technical Reports Server (NTRS)

Rakoczy, John; Heater, Daniel; Lee, Ashley

2013-01-01

Marshall Space Flight Center's (MSFC) Small Projects Rapid Integration and Test Environment (SPRITE) is a Hardware-In-The-Loop (HWIL) facility that provides rapid development, integration, and testing capabilities for small projects (CubeSats, payloads, spacecraft, and launch vehicles). This facility environment focuses on efficient processes and modular design to support rapid prototyping, integration, testing and verification of small projects at an affordable cost, especially compared to larger type HWIL facilities. SPRITE (Figure 1) consists of a "core" capability or "plant" simulation platform utilizing a graphical programming environment capable of being rapidly re-configured for any potential test article's space environments, as well as a standard set of interfaces (i.e. Mil-Std 1553, Serial, Analog, Digital, etc.). SPRITE also allows this level of interface testing of components and subsystems very early in a program, thereby reducing program risk.

Universal accessibility of "accessible" fitness and recreational facilities for persons with mobility disabilities.

PubMed

Arbour-Nicitopoulos, Kelly P; Ginis, Kathleen A Martin

2011-01-01

This study descriptively measured the universal accessibility of "accessible" fitness and recreational facilities for Ontarians living with mobility disabilities. The physical and social environments of 44 fitness and recreational facilities that identified as "accessible" were assessed using a modified version of the AIMFREE. None of the 44 facilities were completely accessible. Mean accessibility ratings ranged between 31 and 63 out of a possible 100. Overall, recreational facilities had higher accessibility scores than fitness centers, with significant differences found on professional support and training, entrance areas, and parking lot. A modest correlation was found between the availability of fitness programming and the overall accessibility of fitness-center specific facility areas. Overall, the physical and social environments of the 44 fitness and recreational facilities assessed were limited in their accessibility for persons with mobility disabilities. Future efforts should be directed at establishing and meeting universal accessibility guidelines for Canadian physical activity facilities.

40 CFR 160.41 - General.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 23 2010-07-01 2010-07-01 false General. 160.41 Section 160.41 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) PESTICIDE PROGRAMS GOOD LABORATORY PRACTICE STANDARDS Facilities § 160.41 General. Each testing facility shall be of suitable size and...

40 CFR 792.41 - General.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 31 2010-07-01 2010-07-01 true General. 792.41 Section 792.41 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) TOXIC SUBSTANCES CONTROL ACT (CONTINUED) GOOD LABORATORY PRACTICE STANDARDS Facilities § 792.41 General. Each testing facility shall be of...

40 CFR 257.4 - Effective date.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 25 2011-07-01 2011-07-01 false Effective date. 257.4 Section 257.4 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES CRITERIA FOR CLASSIFICATION OF SOLID WASTE DISPOSAL FACILITIES AND PRACTICES Classification of Solid Waste Disposal Facilities...

40 CFR 256.25 - Recommendation for inactive facilities.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 24 2010-07-01 2010-07-01 false Recommendation for inactive facilities. 256.25 Section 256.25 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES GUIDELINES FOR DEVELOPMENT AND IMPLEMENTATION OF STATE SOLID WASTE MANAGEMENT PLANS Solid Waste...

40 CFR 257.4 - Effective date.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 24 2010-07-01 2010-07-01 false Effective date. 257.4 Section 257.4 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES CRITERIA FOR CLASSIFICATION OF SOLID WASTE DISPOSAL FACILITIES AND PRACTICES Classification of Solid Waste Disposal Facilities...

40 CFR 256.25 - Recommendation for inactive facilities.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 25 2011-07-01 2011-07-01 false Recommendation for inactive facilities. 256.25 Section 256.25 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES GUIDELINES FOR DEVELOPMENT AND IMPLEMENTATION OF STATE SOLID WASTE MANAGEMENT PLANS Solid Waste...

40 CFR 113.2 - Applicability.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 23 2012-07-01 2012-07-01 false Applicability. 113.2 Section 113.2 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS LIABILITY LIMITS FOR SMALL ONSHORE STORAGE FACILITIES Oil Storage Facilities § 113.2 Applicability. This subpart applies to...

40 CFR 113.1 - Purpose.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 22 2014-07-01 2013-07-01 true Purpose. 113.1 Section 113.1 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS LIABILITY LIMITS FOR SMALL ONSHORE STORAGE FACILITIES Oil Storage Facilities § 113.1 Purpose. This subpart establishes size...

40 CFR 113.3 - Definitions.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 23 2012-07-01 2012-07-01 false Definitions. 113.3 Section 113.3 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS LIABILITY LIMITS FOR SMALL ONSHORE STORAGE FACILITIES Oil Storage Facilities § 113.3 Definitions. As used in this subpart...

40 CFR 113.1 - Purpose.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 23 2012-07-01 2012-07-01 false Purpose. 113.1 Section 113.1 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS LIABILITY LIMITS FOR SMALL ONSHORE STORAGE FACILITIES Oil Storage Facilities § 113.1 Purpose. This subpart establishes size...

40 CFR 113.5 - Exclusions.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 22 2014-07-01 2013-07-01 true Exclusions. 113.5 Section 113.5 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS LIABILITY LIMITS FOR SMALL ONSHORE STORAGE FACILITIES Oil Storage Facilities § 113.5 Exclusions. This subpart does not apply...

40 CFR 113.3 - Definitions.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 22 2014-07-01 2013-07-01 true Definitions. 113.3 Section 113.3 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS LIABILITY LIMITS FOR SMALL ONSHORE STORAGE FACILITIES Oil Storage Facilities § 113.3 Definitions. As used in this subpart...

40 CFR 113.5 - Exclusions.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 23 2012-07-01 2012-07-01 false Exclusions. 113.5 Section 113.5 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS LIABILITY LIMITS FOR SMALL ONSHORE STORAGE FACILITIES Oil Storage Facilities § 113.5 Exclusions. This subpart does not apply...

40 CFR 113.2 - Applicability.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 22 2014-07-01 2013-07-01 true Applicability. 113.2 Section 113.2 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS LIABILITY LIMITS FOR SMALL ONSHORE STORAGE FACILITIES Oil Storage Facilities § 113.2 Applicability. This subpart applies to...

[Nuclear energy and environment: review of the IAEA environmental projects].

PubMed

Fesenko, S; Fogt, G

2012-01-01

The review of the environmental projects of the International Atomic Energy Agency is presented. Basic IAEA documents intended to protect humans and the Environment are considered and their main features are discussed. Some challenging issues in the area of protection of the Environment and man, including the impact of nuclear facilities on the environment, radioactive waste management, and remediation of the areas affected by radiological accidents, nuclear testing and sites of nuclear facilities are also discussed. The need to maintain the existing knowledge in radioecology and protection of the environment is emphasised.

Embracing Safe Ground Test Facility Operations and Maintenance

NASA Technical Reports Server (NTRS)

Dunn, Steven C.; Green, Donald R.

2010-01-01

Conducting integrated operations and maintenance in wind tunnel ground test facilities requires a balance of meeting due dates, efficient operation, responsiveness to the test customer, data quality, effective maintenance (relating to readiness and reliability), and personnel and facility safety. Safety is non-negotiable, so the balance must be an "and" with other requirements and needs. Pressure to deliver services faster at increasing levels of quality in under-maintained facilities is typical. A challenge for management is to balance the "need for speed" with safety and quality. It s especially important to communicate this balance across the organization - workers, with a desire to perform, can be tempted to cut corners on defined processes to increase speed. Having a lean staff can extend the time required for pre-test preparations, so providing a safe work environment for facility personnel and providing good stewardship for expensive National capabilities can be put at risk by one well-intending person using at-risk behavior. This paper documents a specific, though typical, operational environment and cites management and worker safety initiatives and tools used to provide a safe work environment. Results are presented and clearly show that the work environment is a relatively safe one, though still not good enough to keep from preventing injury. So, the journey to a zero injury work environment - both in measured reality and in the minds of each employee - continues. The intent of this paper is to provide a benchmark for others with operational environments and stimulate additional sharing and discussion on having and keeping a safe work environment.

Comparative Measurements of Earth and Martian Entry Environments in the NASA Langley HYMETS Facility

NASA Technical Reports Server (NTRS)

Splinter, Scott C.; Bey, Kim S.; Gragg, Jeffrey G.; Brewer, Amy

2011-01-01

Arc-jet facilities play a major role in the development of heat shield materials for entry vehicles because they are capable of producing representative high-enthalpy flow environments. Arc-jet test data is used to certify material performance for a particular mission and to validate or calibrate models of material response during atmospheric entry. Materials used on missions entering Earth s atmosphere are certified in an arc-jet using a simulated air entry environment. Materials used on missions entering the Martian atmosphere should be certified in an arc-jet using a simulated Martian atmosphere entry environment, which requires the use of carbon dioxide. Carbon dioxide has not been used as a test gas in a United States arc-jet facility since the early 1970 s during the certification of materials for the Viking Missions. Materials certified for the Viking missions have been used on every entry mission to Mars since that time. The use of carbon dioxide as a test gas in an arc-jet is again of interest to the thermal protection system community for certification of new heat shield materials that can increase the landed mass capability for Mars bound missions beyond that of Viking and Pathfinder. This paper describes the modification, operation, and performance of the Hypersonic Materials Environmental Test System (HYMETS) arc-jet facility with carbon dioxide as a test gas. A basic comparison of heat fluxes, various bulk properties, and performance characteristics for various Earth and Martian entry environments in HYMETS is provided. The Earth and Martian entry environments consist of a standard Earth atmosphere, an oxygen-rich Earth atmosphere, and a simulated Martian atmosphere. Finally, a preliminary comparison of the HYMETS arc-jet facility to several European plasma facilities is made to place the HYMETS facility in a more global context of arc-jet testing capability.

Weightless Environment Training Facility (WETF) materials coating evaluation, volume 2

NASA Technical Reports Server (NTRS)

1995-01-01

This volume consists of Appendices A and B to the report on the Weightless Environment Training Facility Materials Coating Evaluation project. The project selected 10 coating systems to be evaluated in six separate exposure environments, and subject to three tests for physical properties. Appendix A holds the coating system, surface preparation, and application data. Appendix B holds the coating material infrared spectra.

Measuring Work Environment and Performance in Nursing Homes

PubMed Central

Temkin-Greener, Helena; Zheng, Nan (Tracy); Katz, Paul; Zhao, Hongwei; Mukamel, Dana B.

2008-01-01

Background Qualitative studies of the nursing home work environment have long suggested that such attributes as leadership and communication may be related to nursing home performance, including residents' outcomes. However, empirical studies examining these relationships have been scant. Objectives This study is designed to: develop an instrument for measuring nursing home work environment and perceived work effectiveness; test the reliability and validity of the instrument; and identify individual and facility-level factors associated with better facility performance. Research Design and Methods The analysis was based on survey responses provided by managers (N=308) and direct care workers (N=7,418) employed in 162 facilities throughout New York State. Exploratory factor analysis, Chronbach's alphas, analysis of variance, and regression models were used to assess instrument reliability and validity. Multivariate regression models, with fixed facility effects, were used to examine factors associated with work effectiveness. Results The reliability and the validity of the survey instrument for measuring work environment and perceived work effectiveness has been demonstrated. Several individual (e.g. occupation, race) and facility characteristics (e.g. management style, workplace conditions, staffing) that are significant predictors of perceived work effectiveness were identified. Conclusions The organizational performance model used in this study recognizes the multidimensionality of the work environment in nursing homes. Our findings suggest that efforts at improving work effectiveness must also be multifaceted. Empirical findings from such a line of research may provide insights for improving the quality of the work environment and ultimately the quality of residents' care. PMID:19330892

Development of Nano-Liposomal Formulations of Epidermal Growth Factor Receptor Inhibitors and their Pharmacological Interactions on Drug-Sensitive and Drug-Resistant Cancer Cell Lines

NASA Astrophysics Data System (ADS)

Trummer, Brian J.

A rapidly expanding understanding of molecular derangements in cancer cell function has led to the development of selective, targeted chemotherapeutic agents. Growth factor signal transduction networks are frequently activated in an aberrant fashion, particularly through the activity of receptor tyrosine kinases (RTK). This has spurred an intensive effort to develop receptor tyrosine kinase inhibitors (RTKI) that are targeted to specific receptors, or receptor subfamilies. Chapter 1 reviews the pharmacology, preclinical, and clinical aspects of RTKIs that target the epidermal growth factor receptor (EGFR). EGFR inhibitors demonstrate significant success at inhibiting phosphorylation-based signaling pathways that promote cancer cell proliferation. Additionally RTKIs have physicochemical and structural characteristics that enable them to function as inhibitors of multi-drug resistance transport proteins. Thus EGFR inhibitors and other RTKIs have both on-target and off-target activities that could be beneficial in cancer therapy. However, these agents exert a number of side effects, some of which arise from their hydrophobic nature and large in vivo volume of distribution. Side effects of the EGFR inhibitor gefitinib include skin rash, severe myelotoxicity when combined with certain chemotherapeutic agents, and impairment of the blood brain barrier to xenobiotics. Weighing the preclinical and clinical observations with the EGFR inhibitors, we developed the primary overall hypothesis of this research: that drug-carrier formulations of RTKIs such as the EGFR inhibitors could be developed based on nanoparticulate liposomal carriers. Theoretically, this carrier strategy would ameliorate toxicity and improve the biodistribution and tumor selectivity of these agents. We hypothesized specifically that liposomal formulations could shift the biodistribution of EGFR inhibitors such as gefitinib away from skin, bone marrow, and the blood brain barrier, and toward solid tumors, due to leaky tumor vasculature and the resulting Enhanced Permeability and Retention (EPR) phenomenon. In Chapter 2 we report that both gefitinib and the structurally similar EGFR inhibitor erlotinib display environment-dependent fluorescence properties. Peak excitation was 345 nm, and the emission peak ranged from 365 to 476 nm, depending upon the polarity of the environment and physical state of the drug. The fluorescence was negligible in aqueous solution, but intense in organic solvents or membrane bilayers. The environment-sensitive fluorescence properties of these drugs enabled rapid evaluation of numerous parameters affecting liposomal drug incorporation and performance. Up to 4-6 mol% of gefitinib could be incorporated in the liposome bilayer, based upon hydrophobic interactions with membrane bilayers. In contrast, 40-60 mol% could be loaded into the aqueous core of pre-formed liposomes at high efficiency, using a remote loading procedure. A stable formulation consisting of distearoylphosphatidylcholine: polyethylene glycol-distereoylphosphatidylethanolamine: cholesterol (DSPC:PEGDSPE:Chol, 9:1:5 mol:mol:mol) and containing drug at 50-60 mol% gefitinib (L-GEF) showed minimal leakage in serum-containing medium over 24 h at 37°C, which should be sufficient to improve biodistribution in vivo. Chapter 3 investigated the pharmacological activity of liposome-encapsulated gefitinib, alone and in combination with several prevalent anticancer agents. Experiments with MCF7 breast cancer cell lines demonstrated that liposome encapsulated gefitinib formulation (L-GEF) had a 2-fold higher IC50 (concentration of drug resulting in half-maximal growth inhibition) than free gefitinib. Lower in vitro potency would be consistent with delayed drug release from the carrier. Therapeutic effects were investigated in combination with the cytotoxic agents paclitaxel and doxorubicin. The drug-resistant MCF7R cell line was 23-fold more resistant to paclitaxel than the parental, drug-sensitive MCF7S cell line, and MCF7R was 12-fold more resistant than MCF7S to doxorubicin. A concentration of 3 muM gefitinib, which had no discernible effect upon MCF7R cell proliferation, reduced resistance to paclitaxel to 6-fold, relative to the parental MCF7S cell line, and reduced resistance to doxorubicin to 8-fold, compared to MCF7S. The cytostatic effect of a wider range of gefitinib:paclitaxel ratios was evaluated in order to permit quantitative analysis of the mechanisms of drug interaction, using response surface models. LGEF in combination with paclitaxel was found to be synergistic in MCF7R multidrug-resistant cells, with a psi value of 0.29. Free GEF combined with paclitaxel was also synergistic, with a psi value of 0.40. The confidence intervals for both psi values were below 1, indicating statistical significance for synergy. The fluorescence of gefitinib and doxorubicin permitted visualization of cellular uptake and intracellular distribution of these drugs in multicellular spheroids of rat 9L gliosarcoma cells, which recapitulate some of the barrier functions to drug distribution within tumors. Gefitinib altered the intracellular distribution of doxorubicin, and overcame the nuclear exclusion of doxorubicin in these drug-resistant cells. Chapter 4 discusses the significance of the findings relating both to the formulations produced and to their pharmacology, and concludes that nanoparticulate formulations of gefitinib have the properties necessary to alter drug biodistribution and pharmacokinetics. Such formulations have the potential to provide an effective means to enhance EGFR-inhibitor efficacy in combination chemotherapy.

40 CFR 792.15 - Inspection of a testing facility.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 31 2010-07-01 2010-07-01 true Inspection of a testing facility. 792.15 Section 792.15 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) TOXIC SUBSTANCES CONTROL ACT (CONTINUED) GOOD LABORATORY PRACTICE STANDARDS General Provisions § 792.15 Inspection...

40 CFR 160.45 - Test system supply facilities.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Test system supply facilities. 160.45 Section 160.45 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) PESTICIDE PROGRAMS... be storage areas, as needed, for feed, nutrients, soils, bedding, supplies, and equipment. Storage...

40 CFR 265.18 - Location standards.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 25 2010-07-01 2010-07-01 false Location standards. 265.18 Section 265.18 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES (CONTINUED... FACILITIES General Facility Standards § 265.18 Location standards. The placement of any hazardous waste in a...

Analytische Geometrie

NASA Astrophysics Data System (ADS)

Kemnitz, Arnfried

Der Grundgedanke der Analytischen Geometrie besteht darin, dass geometrische Untersuchungen mit rechnerischen Mitteln geführt werden. Geometrische Objekte werden dabei durch Gleichungen beschrieben und mit algebraischen Methoden untersucht. Behandelt werden folgende Themen: Koordinatensysteme: Kartesisches Koordinatensystem der Ebene und des Raumes, Polarkoordinatensystem der Ebene, Zusammenhang zwischen kartesischen und Polarkoordinaten; Geraden: Geradengleichungen, Abstände von Geraden; Kreise: Kreisgleichungen, Kreisberechnungen; Kugeln; Kegelschnitte; Ellipsen; Hyperbeln; Parabeln; Anwendungen von Kegelschnitten aus Technik und Mathematik; Vektoren: Definitionen, Addition, Multiplikation, Komponentendarstellung in der Ebene und im Raum, Skalarprodukt, Vektorprodukt. Zu den einzelnen Themenkreisen sind Beispiele aufgeführt. Wichtige Regeln und Gesetze sind durch Umrandung besonders kenntlich gemacht.

Fission yield covariances for JEFF: A Bayesian Monte Carlo method

NASA Astrophysics Data System (ADS)

Leray, Olivier; Rochman, Dimitri; Fleming, Michael; Sublet, Jean-Christophe; Koning, Arjan; Vasiliev, Alexander; Ferroukhi, Hakim

2017-09-01

The JEFF library does not contain fission yield covariances, but simply best estimates and uncertainties. This situation is not unique as all libraries are facing this deficiency, firstly due to the lack of a defined format. An alternative approach is to provide a set of random fission yields, themselves reflecting covariance information. In this work, these random files are obtained combining the information from the JEFF library (fission yields and uncertainties) and the theoretical knowledge from the GEF code. Examples of this method are presented for the main actinides together with their impacts on simple burn-up and decay heat calculations.

Mars mission science operations facilities design

NASA Technical Reports Server (NTRS)

Norris, Jeffrey S.; Wales, Roxana; Powell, Mark W.; Backes, Paul G.; Steinke, Robert C.

2002-01-01

A variety of designs for Mars rover and lander science operations centers are discussed in this paper, beginning with a brief description of the Pathfinder science operations facility and its strengths and limitations. Particular attention is then paid to lessons learned in the design and use of operations facilities for a series of mission-like field tests of the FIDO prototype Mars rover. These lessons are then applied to a proposed science operations facilities design for the 2003 Mars Exploration Rover (MER) mission. Issues discussed include equipment selection, facilities layout, collaborative interfaces, scalability, and dual-purpose environments. The paper concludes with a discussion of advanced concepts for future mission operations centers, including collaborative immersive interfaces and distributed operations. This paper's intended audience includes operations facility and situation room designers and the users of these environments.

40 CFR 60.260 - Applicability and designation of affected facility.

Code of Federal Regulations, 2011 CFR

2011-07-01

... affected facility. 60.260 Section 60.260 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY... Performance for Ferroalloy Production Facilities § 60.260 Applicability and designation of affected facility..., ferrochrome silicon, silvery iron, high-carbon ferrochrome, charge chrome, standard ferromanganese...

40 CFR 60.260 - Applicability and designation of affected facility.

Code of Federal Regulations, 2012 CFR

2012-07-01

... affected facility. 60.260 Section 60.260 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY... Performance for Ferroalloy Production Facilities § 60.260 Applicability and designation of affected facility..., ferrochrome silicon, silvery iron, high-carbon ferrochrome, charge chrome, standard ferromanganese...

Interactive Schematic Integration Within the Propellant System Modeling Environment

NASA Technical Reports Server (NTRS)

Coote, David; Ryan, Harry; Burton, Kenneth; McKinney, Lee; Woodman, Don

2012-01-01

Task requirements for rocket propulsion test preparations of the test stand facilities drive the need to model the test facility propellant systems prior to constructing physical modifications. The Propellant System Modeling Environment (PSME) is an initiative designed to enable increased efficiency and expanded capabilities to a broader base of NASA engineers in the use of modeling and simulation (M&S) technologies for rocket propulsion test and launch mission requirements. PSME will enable a wider scope of users to utilize M&S of propulsion test and launch facilities for predictive and post-analysis functionality by offering a clean, easy-to-use, high-performance application environment.

40 CFR 60.370 - Applicability and designation of affected facility.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 6 2010-07-01 2010-07-01 false Applicability and designation of affected facility. 60.370 Section 60.370 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY... Performance for Lead-Acid Battery Manufacturing Plants § 60.370 Applicability and designation of affected...

Experimental methods for studying microbial survival in extraterrestrial environments.

PubMed

Olsson-Francis, Karen; Cockell, Charles S

2010-01-01

Microorganisms can be used as model systems for studying biological responses to extraterrestrial conditions; however, the methods for studying their response are extremely challenging. Since the first high altitude microbiological experiment in 1935 a large number of facilities have been developed for short- and long-term microbial exposure experiments. Examples are the BIOPAN facility, used for short-term exposure, and the EXPOSE facility aboard the International Space Station, used for long-term exposure. Furthermore, simulation facilities have been developed to conduct microbiological experiments in the laboratory environment. A large number of microorganisms have been used for exposure experiments; these include pure cultures and microbial communities. Analyses of these experiments have involved both culture-dependent and independent methods. This review highlights and discusses the facilities available for microbiology experiments, both in space and in simulation environments. A description of the microorganisms and the techniques used to analyse survival is included. Finally we discuss the implications of microbiological studies for future missions and for space applications. Copyright 2009 Elsevier B.V. All rights reserved.

40 CFR 52.279 - Food processing facilities.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 3 2012-07-01 2012-07-01 false Food processing facilities. 52.279... (CONTINUED) APPROVAL AND PROMULGATION OF IMPLEMENTATION PLANS California § 52.279 Food processing facilities... emissions from food processing facilities without any accompanying analyses demonstrating that these...

40 CFR 52.279 - Food processing facilities.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 3 2014-07-01 2014-07-01 false Food processing facilities. 52.279... (CONTINUED) APPROVAL AND PROMULGATION OF IMPLEMENTATION PLANS California § 52.279 Food processing facilities... emissions from food processing facilities without any accompanying analyses demonstrating that these...

40 CFR 52.279 - Food processing facilities.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 3 2011-07-01 2011-07-01 false Food processing facilities. 52.279... (CONTINUED) APPROVAL AND PROMULGATION OF IMPLEMENTATION PLANS California § 52.279 Food processing facilities... emissions from food processing facilities without any accompanying analyses demonstrating that these...

40 CFR 52.279 - Food processing facilities.

Code of Federal Regulations, 2013 CFR

2013-07-01

... emissions from food processing facilities without any accompanying analyses demonstrating that these... 40 Protection of Environment 3 2013-07-01 2013-07-01 false Food processing facilities. 52.279... (CONTINUED) APPROVAL AND PROMULGATION OF IMPLEMENTATION PLANS California § 52.279 Food processing facilities...

Virtually-augmented interfaces for tactical aircraft.

PubMed

Haas, M W

1995-05-01

The term Fusion Interface is defined as a class of interface which integrally incorporates both virtual and non-virtual concepts and devices across the visual, auditory and haptic sensory modalities. A fusion interface is a multi-sensory virtually-augmented synthetic environment. A new facility has been developed within the Human Engineering Division of the Armstrong Laboratory dedicated to exploratory development of fusion-interface concepts. One of the virtual concepts to be investigated in the Fusion Interfaces for Tactical Environments facility (FITE) is the application of EEG and other physiological measures for virtual control of functions within the flight environment. FITE is a specialized flight simulator which allows efficient concept development through the use of rapid prototyping followed by direct experience of new fusion concepts. The FITE facility also supports evaluation of fusion concepts by operational fighter pilots in a high fidelity simulated air combat environment. The facility was utilized by a multi-disciplinary team composed of operational pilots, human-factors engineers, electronics engineers, computer scientists, and experimental psychologists to prototype and evaluate the first multi-sensory, virtually-augmented cockpit. The cockpit employed LCD-based head-down displays, a helmet-mounted display, three-dimensionally localized audio displays, and a haptic display. This paper will endeavor to describe the FITE facility architecture, some of the characteristics of the FITE virtual display and control devices, and the potential application of EEG and other physiological measures within the FITE facility.

Optimizing the patient-centered environment: results of guided tours with health care providers and employees.

PubMed

Locatelli, Sara M; Turcios, Stephanie; LaVela, Sherri L

2015-01-01

To examine providers' perspectives on the care environment and patient-centered care (PCC) through the eyes of the veteran patient, using guided tours qualitative methodology. Environmental factors, such as attractiveness and function, have the potential to improve patients' experiences. Participatory qualitative methods allow researchers to explore the environment and facilitate discussion. Guided tours were conducted with 25 health care providers/employees at two Veterans Affairs (VA) health care facilities. In guided tours, participants lead the researcher through an environment, commenting on their surroundings, thoughts, and feelings. The researcher walks along with the participant, asking open-ended questions as needed to foster discussion and gain an understanding of the participant's view. Participants were asked to walk through the facility as though they were a veteran. Tours were audio recorded, with participant permission, and transcribed verbatim by research assistants. Three qualitative researchers were responsible for codebook development and coding transcripts and used data-driven coding approaches. Participants discussed physical appearance of the environment and how that influences perceptions about care. Overall, participants highlighted the need to shed the "institutional" appearance. Differences between VA and non-VA health care facilities were discussed, including availability of private rooms and staff to assist with navigating the facility. They reviewed resources in the facility, such as the information desk to assist patients and families. Finally, they offered suggestions for future improvements, including improvements to waiting areas and quiet areas for patients to relax and "get away" from their rooms. Participants highlighted many small changes to the care environment that could enhance the patient experience. Additionally, they examined the environment from the patient's perspective, to identify elements that enhance, or detract from, the patient's care experience. © The Author(s) 2015.

49 CFR 190.233 - Corrective action orders.

Code of Federal Regulations, 2011 CFR

2011-10-01

... facility to be hazardous to life, property, or the environment, the Associate Administrator, OPS shall... the failure to do so would result in the likelihood of serious harm to life, property, or the... Administrator, OPS finds the facility is or would be hazardous to life, property, or the environment, the...

40 CFR 256.63 - Requirements for public participation in the permitting of facilities.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 25 2014-07-01 2014-07-01 false Requirements for public participation in the permitting of facilities. 256.63 Section 256.63 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES GUIDELINES FOR DEVELOPMENT AND IMPLEMENTATION OF STATE SOLID WASTE...

40 CFR 256.63 - Requirements for public participation in the permitting of facilities.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 26 2012-07-01 2011-07-01 true Requirements for public participation in the permitting of facilities. 256.63 Section 256.63 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES GUIDELINES FOR DEVELOPMENT AND IMPLEMENTATION OF STATE SOLID WASTE...

40 CFR 256.63 - Requirements for public participation in the permitting of facilities.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 25 2011-07-01 2011-07-01 false Requirements for public participation in the permitting of facilities. 256.63 Section 256.63 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES GUIDELINES FOR DEVELOPMENT AND IMPLEMENTATION OF STATE SOLID WASTE...

40 CFR 256.63 - Requirements for public participation in the permitting of facilities.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 40 Protection of Environment 26 2013-07-01 2013-07-01 false Requirements for public participation in the permitting of facilities. 256.63 Section 256.63 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES GUIDELINES FOR DEVELOPMENT AND IMPLEMENTATION OF STATE SOLID WASTE...

40 CFR 35.925-1 - Facilities planning.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 1 2012-07-01 2012-07-01 false Facilities planning. 35.925-1 Section 35.925-1 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY GRANTS AND OTHER FEDERAL ASSISTANCE STATE AND LOCAL ASSISTANCE Grants for Construction of Treatment Works-Clean Water Act § 35.925-1...

40 CFR 35.925-1 - Facilities planning.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 1 2014-07-01 2014-07-01 false Facilities planning. 35.925-1 Section 35.925-1 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY GRANTS AND OTHER FEDERAL ASSISTANCE STATE AND LOCAL ASSISTANCE Grants for Construction of Treatment Works-Clean Water Act § 35.925-1...

40 CFR 35.925-1 - Facilities planning.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 40 Protection of Environment 1 2013-07-01 2013-07-01 false Facilities planning. 35.925-1 Section 35.925-1 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY GRANTS AND OTHER FEDERAL ASSISTANCE STATE AND LOCAL ASSISTANCE Grants for Construction of Treatment Works-Clean Water Act § 35.925-1...

40 CFR 35.925-1 - Facilities planning.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 1 2011-07-01 2011-07-01 false Facilities planning. 35.925-1 Section 35.925-1 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY GRANTS AND OTHER FEDERAL ASSISTANCE STATE AND LOCAL ASSISTANCE Grants for Construction of Treatment Works-Clean Water Act § 35.925-1...

49 CFR 190.233 - Corrective action orders.

Code of Federal Regulations, 2010 CFR

2010-10-01

... facility to be hazardous to life, property, or the environment, the Associate Administrator, OPS shall... the failure to do so would result in the likelihood of serious harm to life, property, or the... Administrator, OPS finds the facility is or would be hazardous to life, property, or the environment, the...

40 CFR 35.917-7 - State review and certification of facilities plan.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 1 2010-07-01 2010-07-01 false State review and certification of facilities plan. 35.917-7 Section 35.917-7 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY GRANTS AND OTHER FEDERAL ASSISTANCE STATE AND LOCAL ASSISTANCE Grants for Construction of Treatment Works...

40 CFR 35.917-8 - Submission and approval of facilities plan.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 1 2010-07-01 2010-07-01 false Submission and approval of facilities plan. 35.917-8 Section 35.917-8 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY GRANTS AND OTHER FEDERAL ASSISTANCE STATE AND LOCAL ASSISTANCE Grants for Construction of Treatment Works-Clean...

40 CFR 35.917-9 - Revision or amendment of facilities plan.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 1 2010-07-01 2010-07-01 false Revision or amendment of facilities plan. 35.917-9 Section 35.917-9 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY GRANTS AND OTHER FEDERAL ASSISTANCE STATE AND LOCAL ASSISTANCE Grants for Construction of Treatment Works-Clean...

40 CFR 113.6 - Effect on other laws.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 22 2011-07-01 2011-07-01 false Effect on other laws. 113.6 Section 113.6 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS LIABILITY LIMITS FOR SMALL ONSHORE STORAGE FACILITIES Oil Storage Facilities § 113.6 Effect on other laws. Nothing...

40 CFR 113.6 - Effect on other laws.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 40 Protection of Environment 23 2013-07-01 2013-07-01 false Effect on other laws. 113.6 Section 113.6 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS LIABILITY LIMITS FOR SMALL ONSHORE STORAGE FACILITIES Oil Storage Facilities § 113.6 Effect on other laws. Nothing...

40 CFR 113.6 - Effect on other laws.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 21 2010-07-01 2010-07-01 false Effect on other laws. 113.6 Section 113.6 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS LIABILITY LIMITS FOR SMALL ONSHORE STORAGE FACILITIES Oil Storage Facilities § 113.6 Effect on other laws. Nothing...

40 CFR 264.31 - Design and operation of facility.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 25 2010-07-01 2010-07-01 false Design and operation of facility. 264.31 Section 264.31 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES...-sudden release of hazardous waste or hazardous waste constituents to air, soil, or surface water which...

40 CFR 141.714 - Requirements for uncovered finished water storage facilities.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 23 2014-07-01 2014-07-01 false Requirements for uncovered finished water storage facilities. 141.714 Section 141.714 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for Cryptosporidium Treatment Technique...

40 CFR 113.6 - Effect on other laws.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 22 2014-07-01 2013-07-01 true Effect on other laws. 113.6 Section 113.6 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS LIABILITY LIMITS FOR SMALL ONSHORE STORAGE FACILITIES Oil Storage Facilities § 113.6 Effect on other laws. Nothing...

40 CFR 113.6 - Effect on other laws.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 23 2012-07-01 2012-07-01 false Effect on other laws. 113.6 Section 113.6 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS LIABILITY LIMITS FOR SMALL ONSHORE STORAGE FACILITIES Oil Storage Facilities § 113.6 Effect on other laws. Nothing...

40 CFR 792.31 - Testing facility management.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 31 2010-07-01 2010-07-01 true Testing facility management. 792.31... facility management. For each study, testing facility management shall: (a) Designate a study director as... appropriately tested for identity, strength, purity, stability, and uniformity, as applicable. (e) Assure that...

MIMI: multimodality, multiresource, information integration environment for biomedical core facilities.

PubMed

Szymanski, Jacek; Wilson, David L; Zhang, Guo-Qiang

2009-10-01

The rapid expansion of biomedical research has brought substantial scientific and administrative data management challenges to modern core facilities. Scientifically, a core facility must be able to manage experimental workflow and the corresponding set of large and complex scientific data. It must also disseminate experimental data to relevant researchers in a secure and expedient manner that facilitates collaboration and provides support for data interpretation and analysis. Administratively, a core facility must be able to manage the scheduling of its equipment and to maintain a flexible and effective billing system to track material, resource, and personnel costs and charge for services to sustain its operation. It must also have the ability to regularly monitor the usage and performance of its equipment and to provide summary statistics on resources spent on different categories of research. To address these informatics challenges, we introduce a comprehensive system called MIMI (multimodality, multiresource, information integration environment) that integrates the administrative and scientific support of a core facility into a single web-based environment. We report the design, development, and deployment experience of a baseline MIMI system at an imaging core facility and discuss the general applicability of such a system in other types of core facilities. These initial results suggest that MIMI will be a unique, cost-effective approach to addressing the informatics infrastructure needs of core facilities and similar research laboratories.

ORNL necessary and sufficient standards for environment, safety, and health. Final report of the Identification Team for other industrial, radiological, and non-radiological hazard facilities

DOE Office of Scientific and Technical Information (OSTI.GOV)

NONE

1998-07-01

This Necessary and Sufficient (N and S) set of standards is for Other Industrial, Radiological, and Non-Radiological Hazard Facilities at Oak Ridge National Laboratory (ORNL). These facility classifications are based on a laboratory-wide approach to classify facilities by hazard category. An analysis of the hazards associated with the facilities at ORNL was conducted in 1993. To identify standards appropriate for these Other Industrial, Radiological, and Non-Radiological Hazard Facilities, the activities conducted in these facilities were assessed, and the hazards associated with the activities were identified. A preliminary hazards list was distributed to all ORNL organizations. The hazards identified in priormore » hazard analyses are contained in the list, and a category of other was provided in each general hazard area. A workshop to assist organizations in properly completing the list was held. Completed hazard screening lists were compiled for each ORNL division, and a master list was compiled for all Other Industrial, Radiological Hazard, and Non-Radiological facilities and activities. The master list was compared against the results of prior hazard analyses by research and development and environment, safety, and health personnel to ensure completeness. This list, which served as a basis for identifying applicable environment, safety, and health standards, appears in Appendix A.« less

Rasfonin, a novel 2-pyrone derivative, induces ras-mutated Panc-1 pancreatic tumor cell death in nude mice.

PubMed

Xiao, Z; Li, L; Li, Y; Zhou, W; Cheng, J; Liu, F; Zheng, P; Zhang, Y; Che, Y

2014-05-22

Rasfonin is a novel 2-pyrone derivative reported to induce apoptosis in ras-dependent cells. In this study, its effects on ras-mutated pancreatic cancer cells were investigated in vitro and in vivo. Two human pancreatic cancer cell lines Panc-1 (mutated K-ras) and BxPC-3 (wild-type K-ras) were selected to test the effects of rasfonin on cell proliferation, clone formation, migration and invasion in vitro. Immunoblotting was used to detect the expressions of EGFR-Ras-Raf-MEK-ERK signaling pathway proteins. Ras activity was measured using a pull-down ELISA kit and guanine exchange factor (GEF)/GTPase-activating proteins (GAP) activity was measured by [(3)H]-GDP radiometric ligand binding. For an in vivo study, CD1 nude mice bearing Panc-1 cells were treated with rasfonin or Salirasib (FTS). We found that rasfonin suppressed proliferation more strongly in Panc-1 cells (IC50=5.5 μM) than BxPC-3 cells (IC50=10 μM) in vitro. Clone formation, migration and invasion by Panc-1 cells were also reduced by rasfonin. Rasfonin had little effect on the farnesylation of Ras, but it strongly downregulated Ras activity and consequently phosphorylation of c-Raf/MEK/ERK. Further experiments indicated that rasfonin reduced Son of sevenless (Sos1) expression but did not alter GEF and GAP activities. The in vivo experiments also revealed that rasfonin (30 mg/kg) delayed the growth of xenograft tumors originating from Panc-1 cells. Tumor weight was ultimately decreased after 20 days of treatment of rasfonin. Rasfonin is a robust inhibitor of pancreatic cancers with the K-ras mutation. The reduction of Sos1 expression and the consequently depressed Ras-MAPK activity could be important in its anticancer activity.

Potent and Selective Peptide-based Inhibition of the G Protein Gαq*

PubMed Central

Charpentier, Thomas H.; Waldo, Gary L.; Lowery-Gionta, Emily G.; Krajewski, Krzysztof; Strahl, Brian D.; Kash, Thomas L.; Harden, T. Kendall; Sondek, John

2016-01-01

In contrast to G protein-coupled receptors, for which chemical and peptidic inhibitors have been extensively explored, few compounds are available that directly modulate heterotrimeric G proteins. Active Gαq binds its two major classes of effectors, the phospholipase C (PLC)-β isozymes and Rho guanine nucleotide exchange factors (RhoGEFs) related to Trio, in a strikingly similar fashion: a continuous helix-turn-helix of the effectors engages Gαq within its canonical binding site consisting of a groove formed between switch II and helix α3. This information was exploited to synthesize peptides that bound active Gαq in vitro with affinities similar to full-length effectors and directly competed with effectors for engagement of Gαq. A representative peptide was specific for active Gαq because it did not bind inactive Gαq or other classes of active Gα subunits and did not inhibit the activation of PLC-β3 by Gβ1γ2. In contrast, the peptide robustly prevented activation of PLC-β3 or p63RhoGEF by Gαq; it also prevented G protein-coupled receptor-promoted neuronal depolarization downstream of Gαq in the mouse prefrontal cortex. Moreover, a genetically encoded form of this peptide flanked by fluorescent proteins inhibited Gαq-dependent activation of PLC-β3 at least as effectively as a dominant-negative form of full-length PLC-β3. These attributes suggest that related, cell-penetrating peptides should effectively inhibit active Gαq in cells and that these and genetically encoded sequences may find application as molecular probes, drug leads, and biosensors to monitor the spatiotemporal activation of Gαq in cells. PMID:27742837

Potent and Selective Peptide-based Inhibition of the G Protein Gαq.

PubMed

Charpentier, Thomas H; Waldo, Gary L; Lowery-Gionta, Emily G; Krajewski, Krzysztof; Strahl, Brian D; Kash, Thomas L; Harden, T Kendall; Sondek, John

2016-12-02

In contrast to G protein-coupled receptors, for which chemical and peptidic inhibitors have been extensively explored, few compounds are available that directly modulate heterotrimeric G proteins. Active Gα q binds its two major classes of effectors, the phospholipase C (PLC)-β isozymes and Rho guanine nucleotide exchange factors (RhoGEFs) related to Trio, in a strikingly similar fashion: a continuous helix-turn-helix of the effectors engages Gα q within its canonical binding site consisting of a groove formed between switch II and helix α3. This information was exploited to synthesize peptides that bound active Gα q in vitro with affinities similar to full-length effectors and directly competed with effectors for engagement of Gα q A representative peptide was specific for active Gα q because it did not bind inactive Gα q or other classes of active Gα subunits and did not inhibit the activation of PLC-β3 by Gβ 1 γ 2 In contrast, the peptide robustly prevented activation of PLC-β3 or p63RhoGEF by Gα q ; it also prevented G protein-coupled receptor-promoted neuronal depolarization downstream of Gα q in the mouse prefrontal cortex. Moreover, a genetically encoded form of this peptide flanked by fluorescent proteins inhibited Gα q -dependent activation of PLC-β3 at least as effectively as a dominant-negative form of full-length PLC-β3. These attributes suggest that related, cell-penetrating peptides should effectively inhibit active Gα q in cells and that these and genetically encoded sequences may find application as molecular probes, drug leads, and biosensors to monitor the spatiotemporal activation of Gα q in cells. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

Cross-species antiviral activity of goose interferon lambda against duck plague virus is related to its positive self-regulatory feedback loop.

PubMed

Chen, Shun; Zhang, Wei; Zhou, Qin; Wang, Anqi; Sun, Lipei; Wang, Mingshu; Jia, Renyong; Zhu, Dekang; Liu, Mafeng; Sun, Kunfeng; Yang, Qiao; Wu, Ying; Chen, Xiaoyue; Cheng, Anchun

2017-06-01

Duck plague virus (DPV) is a virus of the Herpesviridae family that leads to acute disease with a high mortality rate in ducks. Control of the disease contributes to the development of poultry breeding. Type III IFN family (IFN-λs) is a novel member of the IFN family, and goose IFN-λ (goIFN-λ) is a newly identified gene whose antiviral function has only been investigated to a limited extent. Here, the cross-species antiviral activity of goIFN-λ against DPV in duck embryo fibroblasts (DEFs) was studied. We found that pre-treatment with goIFN-λ greatly increased the expression of IFN-λ in both heterologous DEFs and homologous goose embryo fibroblasts (GEFs), while differentially inducing IFNα- and IFN-stimulated genes. Additionally, a positive self-regulatory feedback loop of goIFN-λ was blocked by a mouse anti-goIFN-λ polyclonal antibody, which was confirmed in both homologous GEFs and goose peripheral blood mononuclear cells (PBMCs). The suppression of the BAC-DPV-EGFP by goIFN-λ in DEFs was confirmed by fluorescence microscopy, flow cytometry (FCM) analysis, viral copies and titre detection, which can be rescued by mouse anti-goIFN-λ polyclonal antibody incubation. Finally, reporter gene assays indicated that the cross-species antiviral activity of goIFN-λ against BAC-DPV-EGFP is related to its positive self-regulatory feedback loop and subsequent ISG induction. Our data shed light on the fundamental mechanisms of goIFN-λ antiviral function in vitro and extend the considerable range of therapeutic applications in multiple-poultry disease.

Structure and properties of the anions MF4-, MCl4- and MBr4- (M = C, Si, Ge)

NASA Astrophysics Data System (ADS)

Grein, Friedrich

2015-04-01

Density functional theory (DFT), Møller-Plesset (MP2) and coupled cluster with single and double substitutions including non-iterative triple excitations (CCSD(T)) calculations on the anions MX4-, with M = C, Si, Ge and X = F, Cl, Br, show that GeF4-, SiCl4-, GeCl4- and SiBr4- prefer a C2v conformation, but CCl4- is an elongated C3v structure. CBr4- has Td symmetry in MP2, but is slightly more stable in elongated C3v form with DFT and CCSD(T). GeBr4- has Td symmetry. CF4- and SiF4- are unstable with respect to loss of an electron. Vertical electron affinities (EAs) are negative also for CCl4 and SiCl4, and close to zero for GeF4 and SiBr4. Adiabatic EAs range from 0.47 eV for SiCl4 to 1.78 eV for GeBr4. The lowest excited states at Td symmetry are 2T2 resonances with energies of 2.1-3.5 eV, resulting from excitation of the a1 singly occupied molecular orbital to vacant t2 orbitals. Vertical excitation energies (VEEs) and vibrational frequencies are given for the most stable anionic geometries. Comparison with experimental VEEs for CCl4- is made. From dissociation energies of MX4, MX4-, MX3 and MX3-, appearance energies of X-, MX3-, X2- and MX2- were calculated. Most were found to be in reasonable agreement with experimental values. Theoretical spin densities and g-factors have been compared with experimental results available for CCl4-, SiCl4- and GeCl4-.

Wsc1 and Mid2 Are Cell Surface Sensors for Cell Wall Integrity Signaling That Act through Rom2, a Guanine Nucleotide Exchange Factor for Rho1

PubMed Central

Philip, Bevin; Levin, David E.

2001-01-01

Wsc1 and Mid2 are highly O-glycosylated cell surface proteins that reside in the plasma membrane of Saccharomyces cerevisiae. They have been proposed to function as mechanosensors of cell wall stress induced by wall remodeling during vegetative growth and pheromone-induced morphogenesis. These proteins are required for activation of the cell wall integrity signaling pathway that consists of the small G-protein Rho1, protein kinase C (Pkc1), and a mitogen-activated protein kinase cascade. We show here by two-hybrid experiments that the C-terminal cytoplasmic domains of Wsc1 and Mid2 interact with Rom2, a guanine nucleotide exchange factor (GEF) for Rho1. At least with regard to Wsc1, this interaction is mediated by the Rom2 N-terminal domain. This domain is distinct from the Rho1-interacting domain, suggesting that the GEF can interact simultaneously with a sensor and with Rho1. We also demonstrate that extracts from wsc1 and mid2 mutants are deficient in the ability to catalyze GTP loading of Rho1 in vitro, providing evidence that the function of the sensor-Rom2 interaction is to stimulate nucleotide exchange toward this G-protein. In a related line of investigation, we identified the PMT2 gene in a genetic screen for mutations that confer an additive cell lysis defect with a wsc1 null allele. Pmt2 is a member of a six-protein family in yeast that catalyzes the first step in O mannosylation of target proteins. We demonstrate that Mid2 is not mannosylated in a pmt2 mutant and that this modification is important for signaling by Mid2. PMID:11113201

Gα12 structural determinants of Hsp90 interaction are necessary for serum response element-mediated transcriptional activation.

PubMed

Montgomery, Ellyn R; Temple, Brenda R S; Peters, Kimberly A; Tolbert, Caitlin E; Booker, Brandon K; Martin, Joseph W; Hamilton, Tyler P; Tagliatela, Alicia C; Smolski, William C; Rogers, Stephen L; Jones, Alan M; Meigs, Thomas E

2014-04-01

The G12/13 class of heterotrimeric G proteins, comprising the α-subunits Gα12 and Gα13, regulates multiple aspects of cellular behavior, including proliferation and cytoskeletal rearrangements. Although guanine nucleotide exchange factors for the monomeric G protein Rho (RhoGEFs) are well characterized as effectors of this G protein class, a variety of other downstream targets has been reported. To identify Gα12 determinants that mediate specific protein interactions, we used a structural and evolutionary comparison between the G12/13, Gs, Gi, and Gq classes to identify "class-distinctive" residues in Gα12 and Gα13. Mutation of these residues in Gα12 to their deduced ancestral forms revealed a subset necessary for activation of serum response element (SRE)-mediated transcription, a G12/13-stimulated pathway implicated in cell proliferative signaling. Unexpectedly, this subset of Gα12 mutants showed impaired binding to heat-shock protein 90 (Hsp90) while retaining binding to RhoGEFs. Corresponding mutants of Gα13 exhibited robust SRE activation, suggesting a Gα12-specific mechanism, and inhibition of Hsp90 by geldanamycin or small interfering RNA-mediated lowering of Hsp90 levels resulted in greater downregulation of Gα12 than Gα13 signaling in SRE activation experiments. Furthermore, the Drosophila G12/13 homolog Concertina was unable to signal to SRE in mammalian cells, and Gα12:Concertina chimeras revealed Gα12-specific determinants of SRE activation within the switch regions and a C-terminal region. These findings identify Gα12 determinants of SRE activation, implicate Gα12:Hsp90 interaction in this signaling mechanism, and illuminate structural features that arose during evolution of Gα12 and Gα13 to allow bifurcated mechanisms of signaling to a common cell proliferative pathway.

RUTBC1 Functions as a GTPase-activating Protein for Rab32/38 and Regulates Melanogenic Enzyme Trafficking in Melanocytes*

PubMed Central

Marubashi, Soujiro; Shimada, Hikaru; Fukuda, Mitsunori; Ohbayashi, Norihiko

2016-01-01

Two cell type-specific Rab proteins, Rab32 and Rab38 (Rab32/38), have been proposed as regulating the trafficking of melanogenic enzymes, including tyrosinase and tyrosinase-related protein 1 (Tyrp1), to melanosomes in melanocytes. Like other GTPases, Rab32/38 function as switch molecules that cycle between a GDP-bound inactive form and a GTP-bound active form; the cycle is thought to be regulated by an activating enzyme, guanine nucleotide exchange factor (GEF), and an inactivating enzyme, GTPase-activating protein (GAP), which stimulates the GTPase activity of Rab32/38. Although BLOC-3 has already been identified as a Rab32/38-specific GEF that regulates the trafficking of tyrosinase and Tyrp1, no physiological GAP for Rab32/38 in melanocytes has ever been identified, and it has remained unclear whether Rab32/38 is involved in the trafficking of dopachrome tautomerase, another melanogenic enzyme, in mouse melanocytes. In this study we investigated RUTBC1, which was originally characterized as a Rab9-binding protein and GAP for Rab32 and Rab33B in vitro, and the results demonstrated that RUTBC1 functions as a physiological GAP for Rab32/38 in the trafficking of all three melanogenic enzymes in mouse melanocytes. The results of this study also demonstrated the involvement of Rab9A in the regulation of the RUTBC1 localization and in the trafficking of all three melanogenic enzymes. We discovered that either excess activation or inactivation of Rab32/38 achieved by manipulating RUTBC1 inhibits the trafficking of all three melanogenic enzymes. These results collectively indicate that proper spatiotemporal regulation of Rab32/38 is essential for the trafficking of all three melanogenic enzymes in mouse melanocytes. PMID:26620560

[Functional analysis of Oct4 promoter in Xuhuai goat].

PubMed

Wei, Guanghui; Li, Dong; Zuo, Qisheng; Zhang, Yani; Zhu, Rui; Zhang, Lei; Liu, Zhiyong; Qiu, Fenglong; Li, Bichun

2014-08-01

The aim of this study was to determine the activity region of Oct4 (octamer-binding transcription factor 4) promoter in Xuhuai goat, and to investigate the effect of TSA (trichostatin A) and VPA(valproicacid) on Oct4 promoter activity. Specific PCR primers of Oct4 promoter including different lengths of fragments were designed by Primer 5.0, then were amplified and cloned into PGL3-Bacic luciferase reporter vector. All the reconstruction vectors were transfected into gEF, P19 and COS7 cells, respectively. After TSA and VPA treatment, the activity of dual-luciferase reporter gene in these three transfected cells was detected. In addition, the CMV promoter of pEGFP-N1 was replaced by the -1516─+30 bp fragment of Oct4 promoter, GFP fluorescence was used to detect the activity of Oct4 promoter. The results indicated that different fragments of Oct4 promoter showed different degrees of activity in gEF, P19 and COS7 cells, and the maximal activity region of Oct4 promoter was -1516─+30 bp, the basal activity region was -238─+30 bp. Positive regulatory domains existed in the region of -1516─-946 bp and -615─-96 bp, while negative regulatory domains existed in the region of -1936─-1516 bp and -946─-615 bp. The optimum induction concentration to enhance the activity of Oct4 promoter was 1 μmol/L of TSA and 4 mmol/L of VPA. The GFP expression can be started by the fragment of -1516─+30 bp. This study provides an experimental basis for revealing the mechanism of expression and regulation of Oct4 in goat.

The guanine nucleotide exchange factor Ric-8A induces domain separation and Ras domain plasticity in Gαi1

PubMed Central

Van Eps, Ned; Thomas, Celestine J.; Hubbell, Wayne L.; Sprang, Stephen R.

2015-01-01

Heterotrimeric G proteins are activated by exchange of GDP for GTP at the G protein alpha subunit (Gα), most notably by G protein-coupled transmembrane receptors. Ric-8A is a soluble cytoplasmic protein essential for embryonic development that acts as both a guanine nucleotide exchange factor (GEF) and a chaperone for Gα subunits of the i, q, and 12/13 classes. Previous studies demonstrated that Ric-8A stabilizes a dynamically disordered state of nucleotide-free Gα as the catalytic intermediate for nucleotide exchange, but no information was obtained on the structures involved or the magnitude of the structural fluctuations. In the present study, site-directed spin labeling (SDSL) together with double electron-electron resonance (DEER) spectroscopy is used to provide global distance constraints that identify discrete members of a conformational ensemble in the Gαi1:Ric-8A complex and the magnitude of structural differences between them. In the complex, the helical and Ras-like nucleotide-binding domains of Gαi1 pivot apart to occupy multiple resolved states with displacements as large as 25 Å. The domain displacement appears to be distinct from that observed in Gαs upon binding of Gs to the β2 adrenergic receptor. Moreover, the Ras-like domain exhibits structural plasticity within and around the nucleotide-binding cavity, and the switch I and switch II regions, which are known to adopt different conformations in the GDP- and GTP-bound states of Gα, undergo structural rearrangements. Collectively, the data show that Ric-8A induces a conformationally heterogeneous state of Gαi and provide insight into the mechanism of action of a nonreceptor Gα GEF. PMID:25605908

Structural diversities induced by cation sizes in a series of fluorogermanophosphates: A2[GeF2(HPO4)2] (A = Na, K, Rb, NH4, and Cs).

PubMed

Chen, Zhang-Gai; Huang, Xia; Zhuang, Rong-Chuan; Zhang, Yu; Liu, Xin; Shi, Tao; Wang, Shuai-Hua; Wu, Shao-Fan; Mi, Jin-Xiao; Huang, Ya-Xi

2017-09-12

Germanophosphates, in comparison with other metal phosphates, have been less studied but potentially exhibit more diverse structural chemistry with wide applications. Herein we applied a hydro-/solvo-fluorothermal route to make use of both the "tailor effect" of fluoride for the formation of low dimensional anionic clusters and the presence of alkali cations of different sizes to align the anionic clusters to control the overall crystal symmetries of germanophosphates. The synergetic effects of fluoride and alkali cations led to structural changes from chain-like structures to layered structures in a series of five novel fluorogermanophosphates: A 2 [GeF 2 (HPO 4 ) 2 ] (A = Na, K, Rb, NH 4 , and Cs, denoted as Na, K, Rb, NH4, and Cs). Although these fluorogermanophosphates have stoichiometrically equivalent formulas, they feature different anionic clusters, diverse structural dimensionalities, and contrasting crystal symmetries. Chain-like structures were observed for the compounds with the smaller sized alkali ions (Na + , K + , and Rb + ), whereas layered structures were found for those containing the larger sized cations ((NH 4 ) + and Cs + ). Specifically, monoclinic space groups were observed for the Na, K, Rb, and NH4 compounds, whereas a tetragonal space group P4/mbm was found for the Cs compound. These compounds provide new insights into the effects of cation sizes on the anionic clusters built from GeO 4 F 2 octahedra and HPO 4 tetrahedra as well as their influences on the overall structural symmetries in germanophosphates. Further characterization including IR spectroscopy and thermal analyses for all five compounds is also presented.

Revisiting the Roco G-protein cycle.

PubMed

Terheyden, Susanne; Ho, Franz Y; Gilsbach, Bernd K; Wittinghofer, Alfred; Kortholt, Arjan

2015-01-01

Mutations in leucine-rich-repeat kinase 2 (LRRK2) are the most frequent cause of late-onset Parkinson's disease (PD). LRRK2 belongs to the Roco family of proteins which share a conserved Ras-like G-domain (Roc) and a C-terminal of Roc (COR) domain tandem. The nucleotide state of small G-proteins is strictly controlled by guanine-nucleotide-exchange factors (GEFs) and GTPase-activating proteins (GAPs). Because of contradictory structural and biochemical data, the regulatory mechanism of the LRRK2 Roc G-domain and the RocCOR tandem is still under debate. In the present study, we solved the first nucleotide-bound Roc structure and used LRRK2 and bacterial Roco proteins to characterize the RocCOR function in more detail. Nucleotide binding induces a drastic structural change in the Roc/COR domain interface, a region strongly implicated in patients with an LRRK2 mutation. Our data confirm previous assumptions that the C-terminal subdomain of COR functions as a dimerization device. We show that the dimer formation is independent of nucleotide. The affinity for GDP/GTP is in the micromolar range, the result of which is high dissociation rates in the s-1 range. Thus Roco proteins are unlikely to need GEFs to achieve activation. Monomeric LRRK2 and Roco G-domains have a similar low GTPase activity to small G-proteins. We show that GTPase activity in bacterial Roco is stimulated by the nucleotide-dependent dimerization of the G-domain within the complex. We thus propose that the Roco proteins do not require GAPs to stimulate GTP hydrolysis but stimulate each other by one monomer completing the catalytic machinery of the other.

eIF2β is critical for eIF5-mediated GDP-dissociation inhibitor activity and translational control

PubMed Central

Jennings, Martin D.; Kershaw, Christopher J.; White, Christopher; Hoyle, Danielle; Richardson, Jonathan P.; Costello, Joseph L.; Donaldson, Ian J.; Zhou, Yu; Pavitt, Graham D.

2016-01-01

In protein synthesis translation factor eIF2 binds initiator tRNA to ribosomes and facilitates start codon selection. eIF2 GDP/GTP status is regulated by eIF5 (GAP and GDI functions) and eIF2B (GEF and GDF activities), while eIF2α phosphorylation in response to diverse signals is a major point of translational control. Here we characterize a growth suppressor mutation in eIF2β that prevents eIF5 GDI and alters cellular responses to reduced eIF2B activity, including control of GCN4 translation. By monitoring the binding of fluorescent nucleotides and initiator tRNA to purified eIF2 we show that the eIF2β mutation does not affect intrinsic eIF2 affinities for these ligands, neither does it interfere with eIF2 binding to 43S pre-initiation complex components. Instead we show that the eIF2β mutation prevents eIF5 GDI stabilizing nucleotide binding to eIF2, thereby altering the off-rate of GDP from eIF2•GDP/eIF5 complexes. This enables cells to grow with reduced eIF2B GEF activity but impairs activation of GCN4 targets in response to amino acid starvation. These findings provide support for the importance of eIF5 GDI activity in vivo and demonstrate that eIF2β acts in concert with eIF5 to prevent premature release of GDP from eIF2γ and thereby ensure tight control of protein synthesis initiation. PMID:27458202

eIF2β is critical for eIF5-mediated GDP-dissociation inhibitor activity and translational control.

PubMed

Jennings, Martin D; Kershaw, Christopher J; White, Christopher; Hoyle, Danielle; Richardson, Jonathan P; Costello, Joseph L; Donaldson, Ian J; Zhou, Yu; Pavitt, Graham D

2016-11-16

In protein synthesis translation factor eIF2 binds initiator tRNA to ribosomes and facilitates start codon selection. eIF2 GDP/GTP status is regulated by eIF5 (GAP and GDI functions) and eIF2B (GEF and GDF activities), while eIF2α phosphorylation in response to diverse signals is a major point of translational control. Here we characterize a growth suppressor mutation in eIF2β that prevents eIF5 GDI and alters cellular responses to reduced eIF2B activity, including control of GCN4 translation. By monitoring the binding of fluorescent nucleotides and initiator tRNA to purified eIF2 we show that the eIF2β mutation does not affect intrinsic eIF2 affinities for these ligands, neither does it interfere with eIF2 binding to 43S pre-initiation complex components. Instead we show that the eIF2β mutation prevents eIF5 GDI stabilizing nucleotide binding to eIF2, thereby altering the off-rate of GDP from eIF2•GDP/eIF5 complexes. This enables cells to grow with reduced eIF2B GEF activity but impairs activation of GCN4 targets in response to amino acid starvation. These findings provide support for the importance of eIF5 GDI activity in vivo and demonstrate that eIF2β acts in concert with eIF5 to prevent premature release of GDP from eIF2γ and thereby ensure tight control of protein synthesis initiation. © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

Role of Abl kinase and the Wave2 signaling complex in HIV-1 entry at a post-hemifusion step.

PubMed

Harmon, Brooke; Campbell, Nancy; Ratner, Lee

2010-06-17

Entry of human immunodeficiency virus type 1 (HIV-1) commences with binding of the envelope glycoprotein (Env) to the receptor CD4, and one of two coreceptors, CXCR4 or CCR5. Env-mediated signaling through coreceptor results in Galphaq-mediated Rac activation and actin cytoskeleton rearrangements necessary for fusion. Guanine nucleotide exchange factors (GEFs) activate Rac and regulate its downstream protein effectors. In this study we show that Env-induced Rac activation is mediated by the Rac GEF Tiam-1, which associates with the adaptor protein IRSp53 to link Rac to the Wave2 complex. Rac and the tyrosine kinase Abl then activate the Wave2 complex and promote Arp2/3-dependent actin polymerization. Env-mediated cell-cell fusion, virus-cell fusion and HIV-1 infection are dependent on Tiam-1, Abl, IRSp53, Wave2, and Arp3 as shown by attenuation of fusion and infection in cells expressing siRNA targeted to these signaling components. HIV-1 Env-dependent cell-cell fusion, virus-cell fusion and infection were also inhibited by Abl kinase inhibitors, imatinib, nilotinib, and dasatinib. Treatment of cells with Abl kinase inhibitors did not affect cell viability or surface expression of CD4 and CCR5. Similar results with inhibitors and siRNAs were obtained when Env-dependent cell-cell fusion, virus-cell fusion or infection was measured, and when cell lines or primary cells were the target. Using membrane curving agents and fluorescence microscopy, we showed that inhibition of Abl kinase activity arrests fusion at the hemifusion (lipid mixing) step, suggesting a role for Abl-mediated actin remodeling in pore formation and expansion. These results suggest a potential utility of Abl kinase inhibitors to treat HIV-1 infected patients.

An oxidation and erosion test facility for cooled panels

NASA Technical Reports Server (NTRS)

Swartwout, W. H.; Erdos, J. I.; Engers, R. J.; Prescott, C.

1992-01-01

The Panel Oxidation and Erosion Testbed (POET) facility under construction at GASL to provide the required test environment is described. The POET facility comprises three major element including a vitiated air heater, a supersonic nozzle, and a test section. A hydrogen-fueld vitiated air heater will provide the oxidizing and erosive environment. The flow through the test section characterized by low supersonic speed and Mach number of 1.4 will maximize the local heat transfer rate and the local surface shear stress.

Extreme Environments Capabilities at Glenn Research Center

NASA Technical Reports Server (NTRS)

Balcerski, Jeffrey; Kremic, Tibor; Arnett, Lori; Vento, Dan; Nakley, Leah

2016-01-01

The NASA Glenn Research Center has several facilities that can provide testing for extreme evironments of interest to the New Frontiers community. This includes the Glenn Extreme Enivironments Rig (GEER) which can duplicate the atmospheric chemistry and conditions for the Venus surface or any other planet with a hot environment. GRC also has several cryogenic facilities which have the capability to run with hydrogen atmospheres, hydrocarbon atmosphere, CO2 based atmospheres or nitrogen atmospheres. The cryogenic facilities have the capability to emulate Titan lakes.

NASA Lighting Research, Test, & Analysis

NASA Technical Reports Server (NTRS)

Clark, Toni

2015-01-01

The Habitability and Human Factors Branch, at Johnson Space Center, in Houston, TX, provides technical guidance for the development of spaceflight lighting requirements, verification of light system performance, analysis of integrated environmental lighting systems, and research of lighting-related human performance issues. The Habitability & Human Factors Lighting Team maintains two physical facilities that are integrated to provide support. The Lighting Environment Test Facility (LETF) provides a controlled darkroom environment for physical verification of lighting systems with photometric and spetrographic measurement systems. The Graphics Research & Analysis Facility (GRAF) maintains the capability for computer-based analysis of operational lighting environments. The combined capabilities of the Lighting Team at Johnson Space Center have been used for a wide range of lighting-related issues.

40 CFR 160.31 - Testing facility management.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Testing facility management. 160.31... GOOD LABORATORY PRACTICE STANDARDS Organization and Personnel § 160.31 Testing facility management. For each study, testing facility management shall: (a) Designate a study director as described in § 160.33...

40 CFR 63.11166 - What General Provisions apply to primary beryllium production facilities?

Code of Federal Regulations, 2010 CFR

2010-07-01

... Primary Nonferrous Metals Area Sources-Zinc, Cadmium, and Beryllium Primary Beryllium Production Facilities § 63.11166 What General Provisions apply to primary beryllium production facilities? (a) You must... primary beryllium production facilities? 63.11166 Section 63.11166 Protection of Environment ENVIRONMENTAL...

Estimate of air carrier and air taxi crash frequencies from high altitude en route flight operations

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sanzo, D.; Kimura, C.Y.; Prassinos, P.G.

1996-06-03

In estimating the frequency of an aircraft crashing into a facility, it has been found convenient to break the problem down into two broad categories. One category estimates the aircraft crash frequency due to air traffic from nearby airports, the so-called near-airport environment. The other category estimates the aircraft crash frequency onto facilities due to air traffic from airways, jet routes, and other traffic flying outside the near-airport environment The total aircraft crash frequency is the summation of the crash frequencies from each airport near the facility under evaluation and from all airways, jet routes, and other traffic near themore » facility of interest. This paper will examine the problems associated with the determining the aircraft crash frequencies onto facilities outside the near-airport environment. This paper will further concentrate on the estimating the risk of aircraft crashes to ground facilities due to high altitude air carrier and air taxi traffic. High altitude air carrier and air taxi traffic will be defined as all air carrier and air taxi flights above 18,000 feet Mean Sea Level (MSL).« less

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ward, Anderson; Basabilvazo, George T.

The purpose of the Waste Isolation Pilot Plant (WIPP) Annual Site Environmental Report for 2016 (ASER) is to provide the information required by U.S. Department of Energy (DOE) Order 231.1B, Environment, Safety, and Health Reporting. The DOE Carlsbad Field Office (CBFO) and the management and operating contractor (MOC) maintain and preserve the environmental resources at the WIPP facility. DOE Order 231.1B; DOE Order 436.1, Departmental Sustainability; and DOE Order 458.1, Radiation Protection of the Public and the Environment, require that the affected environment at and near DOE facilities be monitored to ensure the safety and health of the public andmore » workers, and preservation of the environment. This report was prepared in accordance with DOE Order 231.1B, which requires DOE facilities to submit an ASER to the DOE Headquarters Chief Health, Safety, and Security Officer.« less

Propulsion Ground Testing: Planning for the Future

NASA Technical Reports Server (NTRS)

Bruce, Robert

2003-01-01

Advanced planners are constantly being asked to plan for the provision of future test capability. Historically, this capability is provided either by substantial investment in new test facility capabilities, or in the substantial investment in the modification of pre-exiting test facilities. The key words in the previous sentence are 'substantial investment.' In the evolving environment of increasingly constrained resources, how is an advanced planner to plan for the provisions of such capabilities? Additionally, the conundrum exists that program formulation decisions are being made based on both life cycle cost decisions in an environment in which the more immediate challenge of front-end capital investment oftentimes is the linchpin upon which early decisions are made. In such an environment, how are plans and decisions made? This paper cites examples of decisions made in the past in the area of both major test facility upgrades, as well as major new test facility investment.

The Proliferation of PDC-Type Environments in Industry and Universities

NASA Technical Reports Server (NTRS)

Shishko, R.

2000-01-01

JPL's Project Design Cenger (PDC), opened in 1994, has become a model for other facilities of the same type in the aerospace industry. More recently, PDC-type environments have been adopted by some university aerospace departments as an educational tool. This paper discusses some of these facilities and their possible future direction.

Comparing Two Types of Model Progression in an Inquiry Learning Environment with Modelling Facilities

ERIC Educational Resources Information Center

Mulder, Yvonne G.; Lazonder, Ard W.; de Jong, Ton

2011-01-01

The educational advantages of inquiry learning environments that incorporate modelling facilities are often challenged by students' poor inquiry skills. This study examined two types of model progression as means to compensate for these skill deficiencies. Model order progression (MOP), the predicted optimal variant, gradually increases the…

Risk and Resilience: Girls' Experiences Navigating Space and Relationships in a Secure Residential Facility

ERIC Educational Resources Information Center

Simonsen, Amy E.

2010-01-01

The purpose of this qualitative study was to examine how adolescent girls with diagnosed learning and emotional disabilities described themselves as they negotiated various environments and relationships within a secure residential facility. The goal was to explore how conditions and interactions in these environments promoted both risk and…

40 CFR 60.5380 - What standards apply to centrifugal compressor affected facilities?

Code of Federal Regulations, 2013 CFR

2013-07-01

... 40 Protection of Environment 7 2013-07-01 2013-07-01 false What standards apply to centrifugal compressor affected facilities? 60.5380 Section 60.5380 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Standards of Performance for Crude Oil and Natura...

40 CFR 60.5380 - What standards apply to centrifugal compressor affected facilities?

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 7 2014-07-01 2014-07-01 false What standards apply to centrifugal compressor affected facilities? 60.5380 Section 60.5380 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Standards of Performance for Crude Oil and Natura...

40 CFR 355.10 - Must my facility comply with the emergency planning requirements of this subpart?

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 28 2011-07-01 2011-07-01 false Must my facility comply with the emergency planning requirements of this subpart? 355.10 Section 355.10 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SUPERFUND, EMERGENCY PLANNING, AND COMMUNITY RIGHT-TO-KNOW PROGRAMS...

40 CFR 60.5375 - What standards apply to gas well affected facilities?

Code of Federal Regulations, 2013 CFR

2013-07-01

... 40 Protection of Environment 7 2013-07-01 2013-07-01 false What standards apply to gas well affected facilities? 60.5375 Section 60.5375 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Standards of Performance for Crude Oil and Natural Gas...

40 CFR 60.5375 - What standards apply to gas well affected facilities?

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 7 2014-07-01 2014-07-01 false What standards apply to gas well affected facilities? 60.5375 Section 60.5375 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Standards of Performance for Crude Oil and Natural Gas...

40 CFR 63.11164 - What General Provisions apply to primary zinc production facilities?

Code of Federal Regulations, 2013 CFR

2013-07-01

... 40 Protection of Environment 15 2013-07-01 2013-07-01 false What General Provisions apply to primary zinc production facilities? 63.11164 Section 63.11164 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) NATIONAL EMISSION STANDARDS FOR HAZARDOUS AIR POLLUTANTS FOR SOURCE CATEGORIES (CONTINUED)...

40 CFR 63.11164 - What General Provisions apply to primary zinc production facilities?

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 15 2012-07-01 2012-07-01 false What General Provisions apply to primary zinc production facilities? 63.11164 Section 63.11164 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) NATIONAL EMISSION STANDARDS FOR HAZARDOUS AIR POLLUTANTS FOR SOURCE CATEGORIES (CONTINUED)...

40 CFR 63.11164 - What General Provisions apply to primary zinc production facilities?

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 15 2014-07-01 2014-07-01 false What General Provisions apply to primary zinc production facilities? 63.11164 Section 63.11164 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) NATIONAL EMISSION STANDARDS FOR HAZARDOUS AIR POLLUTANTS FOR SOURCE CATEGORIES (CONTINUED)...

40 CFR 63.11164 - What General Provisions apply to primary zinc production facilities?

Code of Federal Regulations, 2011 CFR

2011-07-01

... primary zinc production facilities? 63.11164 Section 63.11164 Protection of Environment ENVIRONMENTAL... Primary Nonferrous Metals Area Sources-Zinc, Cadmium, and Beryllium Primary Zinc Production Facilities § 63.11164 What General Provisions apply to primary zinc production facilities? (a) If you own or...

40 CFR 63.11164 - What General Provisions apply to primary zinc production facilities?

Code of Federal Regulations, 2010 CFR

2010-07-01

... primary zinc production facilities? 63.11164 Section 63.11164 Protection of Environment ENVIRONMENTAL... Primary Nonferrous Metals Area Sources-Zinc, Cadmium, and Beryllium Primary Zinc Production Facilities § 63.11164 What General Provisions apply to primary zinc production facilities? (a) If you own or...

40 CFR 160.51 - Specimen and data storage facilities.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 25 2012-07-01 2012-07-01 false Specimen and data storage facilities... PROGRAMS GOOD LABORATORY PRACTICE STANDARDS Facilities § 160.51 Specimen and data storage facilities. Space shall be provided for archives, limited to access by authorized personnel only, for the storage and...

40 CFR 160.51 - Specimen and data storage facilities.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 24 2011-07-01 2011-07-01 false Specimen and data storage facilities... PROGRAMS GOOD LABORATORY PRACTICE STANDARDS Facilities § 160.51 Specimen and data storage facilities. Space shall be provided for archives, limited to access by authorized personnel only, for the storage and...

40 CFR 160.51 - Specimen and data storage facilities.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 40 Protection of Environment 25 2013-07-01 2013-07-01 false Specimen and data storage facilities... PROGRAMS GOOD LABORATORY PRACTICE STANDARDS Facilities § 160.51 Specimen and data storage facilities. Space shall be provided for archives, limited to access by authorized personnel only, for the storage and...

40 CFR 160.51 - Specimen and data storage facilities.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 24 2014-07-01 2014-07-01 false Specimen and data storage facilities... PROGRAMS GOOD LABORATORY PRACTICE STANDARDS Facilities § 160.51 Specimen and data storage facilities. Space shall be provided for archives, limited to access by authorized personnel only, for the storage and...

40 CFR 60.270 - Applicability and designation of affected facility.

Code of Federal Regulations, 2012 CFR

2012-07-01

... affected facility. 60.270 Section 60.270 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY... August 17, 1983 § 60.270 Applicability and designation of affected facility. (a) The provisions of this subpart are applicable to the following affected facilities in steel plants that produce carbon, alloy, or...

40 CFR 60.270 - Applicability and designation of affected facility.

Code of Federal Regulations, 2011 CFR

2011-07-01

... affected facility. 60.270 Section 60.270 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY... August 17, 1983 § 60.270 Applicability and designation of affected facility. (a) The provisions of this subpart are applicable to the following affected facilities in steel plants that produce carbon, alloy, or...

40 CFR 35.925-1 - Facilities planning.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 1 2010-07-01 2010-07-01 false Facilities planning. 35.925-1 Section... Facilities planning. That, if the award is for step 2, step 3, or step 2=3 grant assistance, the facilities planning requirements in § 35.917 et seq. have been met. ...

Mental and behavioral health environments: critical considerations for facility design.

PubMed

Shepley, Mardelle McCuskey; Watson, Angela; Pitts, Francis; Garrity, Anne; Spelman, Elizabeth; Kelkar, Janhawi; Fronsman, Andrea

2016-01-01

The purpose of the study was to identify features in the physical environment that are believed to positively impact staff and patients in psychiatric environments and use these features as the foundation for future research regarding the design of mental and behavioral health facilities. Pursuant to a broad literature review that produced an interview script, researchers conducted 19 interviews of psychiatric staff, facility administrators and architects. Interview data were analyzed using the highly structured qualitative data analysis process authored by Lincoln and Guba (1985). Seventeen topics were addressed ranging from the importance of a deinstitutionalized environment to social interaction and autonomy. The interviewees reinforced the controversy that exists around the implications of a deinstitutionalized environment, when the resulting setting diminishes patient and staff safety. Respondents tended to support open nurse stations vs. enclosed stations. Support for access to nature and the provision of an aesthetic environment was strong. Most interviewees asserted that private rooms were highly desirable because lower room density reduces the institutional character of a unit. However, a few interviewees adamantly opposed private rooms because they considered the increased supervision of one patient by another to be a deterrent to self-harm. The need to address smoking rooms in future research received the least support of all topics. Responses of interviews illustrate current opinion regarding best practice in the design of psychiatric facilities. The findings emphasize the need for more substantive research on appropriate physical environments in mental and behavioral health settings. Copyright © 2016 Elsevier Inc. All rights reserved.

The physical environment, activity and interaction in residential care facilities for older people: a comparative case study.

PubMed

Nordin, Susanna; McKee, Kevin; Wallinder, Maria; von Koch, Lena; Wijk, Helle; Elf, Marie

2017-12-01

The physical environment is of particular importance for supporting activities and interactions among older people living in residential care facilities (RCFs) who spend most of their time inside the facility. More knowledge is needed regarding the complex relationships between older people and environmental aspects in long-term care. The present study aimed to explore how the physical environment influences resident activities and interactions at two RCFs by using a mixed-method approach. Environmental assessments were conducted via the Swedish version of the Sheffield Care Environment Assessment Matrix (S-SCEAM), and resident activities, interactions and locations were assessed through an adapted version of the Dementia Care Mapping (DCM). The Observed Emotion Rating Scale (OERS) was used to assess residents' affective states. Field notes and walk-along interviews were also used. Findings indicate that the design of the physical environment influenced the residents' activities and interactions. Private apartments and dining areas showed high environmental quality at both RCFs, whereas the overall layout had lower quality. Safety was highly supported. Despite high environmental quality in general, several factors restricted resident activities. To optimise care for older people, the design process must clearly focus on accessible environments that provide options for residents to use the facility independently. © 2016 The Authors. Scandinavian Journal of Caring Sciences published by John Wiley & Sons Ltd on behalf of Nordic College of Caring Science.

Prevalence of School Policies, Programs, and Facilities That Promote a Healthy Physical School Environment

PubMed Central

Everett Jones, Sherry; Brener, Nancy D.; McManus, Tim

2003-01-01

Objectives. We examined the extent to which schools in the United States have health-promoting policies, programs, and facilities. Methods. We analyzed data from the School Health Policies and Programs Study 2000. Results. We found that public schools (vs private and Catholic schools), urban schools (vs rural and suburban schools), and schools with larger enrollments (vs smaller schools) had more health-promoting policies, programs, and facilities in place. On average, middle schools had 11.0 and middle/junior and high schools had 10.4 out of a possible 18 policies, programs, and facilities. Conclusions. Although some schools had many healthy physical environment features, room for improvement exists. Resources are available to help schools improve their health-promoting policies, programs, and facilities. PMID:12948982

Neighborhood Environment Walkability Scale for Youth (NEWS-Y): reliability and relationship with physical activity.

PubMed

Rosenberg, Dori; Ding, Ding; Sallis, James F; Kerr, Jacqueline; Norman, Gregory J; Durant, Nefertiti; Harris, Sion K; Saelens, Brian E

2009-01-01

To examine the psychometric properties of the Neighborhood Environment Walkability Scale-Youth (NEWS-Y) and explore its associations with context-specific and overall physical activity (PA) among youth. In 2005, parents of children ages 5-11 (n=116), parents of adolescents ages 12-18 (n=171), and adolescents ages 12-18 (n=171) from Boston, Cincinnati, and San Diego, completed NEWS-Y surveys regarding perceived land use mix-diversity, recreation facility availability, pedestrian/automobile traffic safety, crime safety, aesthetics, walking/cycling facilities, street connectivity, land use mix-access, and residential density. A standardized neighborhood environment score was derived. Self-reported activity in the street and in parks, and walking to parks, shops, school, and overall physical activity were assessed. The NEWS-Y subscales had acceptable test-retest reliability (ICC range .56-.87). Being active in a park, walking to a park, walking to shops, and walking to school were related to multiple environmental attributes in all three participant groups. Total neighborhood environment, recreation facilities, walking and cycling facilities, and land use mix-access had the most consistent relationships with specific types of activity. The NEWS-Y has acceptable reliability and subscales were significantly correlated with specific types of youth PA. The NEWS-Y can be used to examine neighborhood environment correlates of youth PA.

[Leben im Eismeer - Tauchuntersuchungen zur Biologie arktischer Meerespflanzen und Meerestiere

PubMed

Lippert; Karsten; Wiencke

2000-01-01

Die Maske wird nochmals auf Dichtigkeit überprüft, der Knoten der Sicherungsleine mit zwei halben Schlägen fixiert, dann rutscht die Taucherin von der Eiskante in das kalte Wasser. Eine halbe Stunde vergeht, bevor ihr Kopf wieder aus dem Eisloch auftaucht und sie ein großes Sammelnetz nach oben reicht, gefüllt mit verschiedenen Arten von Makroalgen. Obwohl noch große Flächen des Kongsfjordes im arktischen Spitzbergen zugefroren sind und das Festland von einer dicken Schneedecke bedeckt ist, hat unter Wasser in den Algenwäldern bereits der Sommer und damit die Saison der Meeresbiologen begonnen.

Segmentierung von Aortenaneurysmen in CTA-Bildern mit dem statistischen Verfahren der Active Appearance Models

NASA Astrophysics Data System (ADS)

Greiner, Katharina; Egger, Jan; Großkopf, Stefan; Kaftan, Jens N.; Dörner, Ralf; Freisleben, Bernd

In diesem Beitrag werden Active Appearance Models (AAMs) zur Segmentierung der äußeren Kontur von Aortenaneurysmen eingesetzt. Diese Aufgabe ist wegen des geringen Kontrastes zum umliegenden Gewebe und des Aufbaus der teils thrombotisierten oder kalzifizierten Gefäßwände im Bereich eines Aneurysmas so komplex, dass sie aufgrund der Vielgestalt der Kontur in CT-Angiographie-Bildern die Verwendung eines statistischen Modells für Form und eingeschlossene Textur rechtfertigt. Für die Evaluation des Verfahrens wurden verschiedene statistische Modelle aus Schichten von neun CTA-Datensätzen trainiert und die Segmentierung anhand von Leave-One-Out-Tests überprüft.

The effectiveness of environment assessment tools to guide refurbishment of Australian residential aged care facilities: A systematic review.

PubMed

Neylon, Samantha; Bulsara, Caroline; Hill, Anne-Marie

2017-06-01

To determine applicability of environment assessment tools in guiding minor refurbishments of Australian residential aged care facilities. Studies conducted in residential aged care settings using assessment tools which address the physical environment were eligible for inclusion in a systematic review. Given these studies are limited, tools which have not yet been utilised in research settings were also included. Tools were analysed using a critical appraisal screen. Forty-three publications met the inclusion criteria. Ten environment assessment tools were identified, of which four addressed all seven minor refurbishment domains of lighting, colour and contrast, sound, flooring, furniture, signage and way finding. Only one had undergone reliability and validity testing. There are four tools which may be suitable to use for minor refurbishment of Australian residential aged care facilities. Data on their reliability, validity and quality are limited. © 2017 AJA Inc.

40 CFR Table 2 to Subpart Fff of... - Nitrogen Oxides Requirements for Affected Facilities

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 8 2010-07-01 2010-07-01 false Nitrogen Oxides Requirements for Affected Facilities 2 Table 2 to Subpart FFF of Part 62 Protection of Environment ENVIRONMENTAL PROTECTION... Before September 20, 1994 Pt. 62, Subpt. FFF, Table 2 Table 2 to Subpart FFF of Part 62—Nitrogen Oxides...

Virtual Civilian Aeromedical Evacuation Sustainment Training Project (V-CAEST)

DTIC Science & Technology

2015-08-01

evacuation liaison team (AELT), and the mobile aeromedical staging facility (MASF). The content covered in the V-CAEST environment therefore covered the...environment was set-up in a large gymnasium building including a mock military plane and Mobile Aeromedical Staging Facility (MASF) located just...staffing exam backhoe scenarios exam infrastructure interface tsunami infrastructure commander telecommunication disrupting commander

NETL- Severe Environment Corrosion Erosion Facility

ScienceCinema

None

2018-01-16

NETL's Severe Environment Corrosion Erosion Facility in Albany studies how new and old materials will stand up to new operating conditions. Work done in the lab supports NETL's oxy-fuel combustion oxidation work, refractory materials stability work, and the fuels program, in particular the hydrogen membrane materials stability work, to determine how best to upgrade existing power plants.

Evaluation of Supported Placements in Integrated Community Environments Project (SPICE). Executive Summary of the Final Report.

ERIC Educational Resources Information Center

Wilson, Leslie; And Others

This executive summary presents highlights of a study which sought to determine whether participants in the Supported Placements in Integrated Community Environments project were better off after moving to community homes from intermediate care facilities and skilled nursing facilities, and to determine the variables that contribute to quality…

Evaluation of Supported Placements in Integrated Community Environments Project (SPICE). Final Report.

ERIC Educational Resources Information Center

Wilson, Leslie; And Others

This evaluation project was designed to assess 37 persons (ages 21-72) who had moved from intermediate care facilities or skilled nursing facilities into innovative one-person or two-person community integrated living arrangements as a result of the Supported Placements in Integrated Community Environments project. The 37 persons had severe or…

Design Principles of an Open Agent Architecture for Web-Based Learning Community.

ERIC Educational Resources Information Center

Jin, Qun; Ma, Jianhua; Huang, Runhe; Shih, Timothy K.

A Web-based learning community involves much more than putting learning materials into a Web site. It can be seen as a complex virtual organization involved with people, facilities, and cyber-environment. Tremendous work and manpower for maintaining, upgrading, and managing facilities and the cyber-environment are required. There is presented an…

Day Care for Persons with Dementia: The Impact of the Physical Environment on Staff Stress and Quality of Care.

ERIC Educational Resources Information Center

Lyman, Karen A.

1989-01-01

Considered impact of physical environment on work-related stress and quality of care in day care center integrating demented people and other frail elderly. Examined positive and negative differences before and after move to new facility. Discusses implications for facility design and other program characteristics. (Author/CM)

Quality of Learning Facilities and Learning Environment: Challenges for Teaching and Learning in Kenya's Public Universities

ERIC Educational Resources Information Center

Ndirangu, Mwangi; Udoto, Maurice O.

2011-01-01

Purpose: The purpose of this article is to report findings on the perceptions of quality of educational facilities in Kenyan public universities, and the implications for teaching/learning, and the learning environment. Design/methodology/approach: The study adopted an exploratory descriptive design. A total of 332 and 107 undergraduate students…

40 CFR 113.4 - Size classes and associated liability limits for fixed onshore oil storage facilities, 1,000...

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 23 2012-07-01 2012-07-01 false Size classes and associated liability limits for fixed onshore oil storage facilities, 1,000 barrels or less capacity. 113.4 Section 113.4 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS LIABILITY LIMITS FOR...

40 CFR 113.4 - Size classes and associated liability limits for fixed onshore oil storage facilities, 1,000...

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 22 2014-07-01 2013-07-01 true Size classes and associated liability limits for fixed onshore oil storage facilities, 1,000 barrels or less capacity. 113.4 Section 113.4 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS LIABILITY LIMITS FOR...

Research Animal Holding Facility Prevents Space Lab Contamination

NASA Technical Reports Server (NTRS)

Savage, P. D., Jr.; Jahns, G. C.; Dalton, B. P.; Hogan, R. P.; Wray, A. E.

1991-01-01

Healthy environment for both rodents and human researchers maintained. Research animal holding facility (RAHF) and rodent cage prevent solid particles (feces, food bits, hair), micro-organisms, ammonia, and odors from escaping into outside environment during spaceflight. Rodent cage contains compartments for two animals. Provides each drinking-water dispenser, feeding alcove, and activity-monitoring port. Feeding and waste trays removable.

Improving children's nutrition environments: A survey of adoption and implementation of nutrition guidelines in recreational facilities

PubMed Central

2011-01-01

Background Although the mandate of recreational facilities is to enhance well-being, many offer foods inconsistent with recommendations for healthy eating. Little is known regarding recreational facility food environments and how they might be improved, as few studies exist. The Alberta Nutrition Guidelines for Children and Youth (ANGCY) are intended to ensure access to healthy food choices in schools, childcare and recreational facilities. This study investigated awareness, adoption and implementation of the ANGCY among recreational facilities in Alberta, Canada, one year following their release. Methods A cross-sectional telephone survey was conducted from June - December, 2009 (n = 151) with managers of publicly funded recreational facilities that served food. The questionnaire included 10 closed and 7 open ended questions to assess the organizational priority for healthy eating, awareness, adoption and implementation of the ANGCY. Chi-squared tests examined quantitative variables, while qualitative data were analysed using directed content analysis. Greenhalgh's model of diffusion of complex innovations within health service organizations constituted the theoretical framework for the study. Results One half of respondents had heard of the ANGCY, however their knowledge of them was limited. Although 51% of facilities had made changes to improve the nutritional quality of foods offered in the past year, only a small fraction (11%) of these changes were motivated by the ANGCY. At the time of the survey, 14% of facilities had adopted the ANGCY and 6% had implemented them. Barriers to adoption and implementation were primarily related to perceived negative attributes of the ANGCY, the inner (organizational) context, and negative feedback received during the implementation process. Managers strongly perceived that implementing nutrition guidelines would limit their profit-making ability. Conclusions If fully adopted and implemented, the ANGCY have the potential to make a significant and sustained contribution to improving the recreational facility food environment, however one year following their release, awareness, adoption and implementation of the ANGCY remained low. A mandated policy approach could offer an efficacious, cost-effective means of improving the food environment within recreational facilities. PMID:21631946

40 CFR 403.19 - Provisions of specific applicability to the Owatonna Waste Water Treatment Facility.

Code of Federal Regulations, 2010 CFR

2010-07-01

... the Owatonna Waste Water Treatment Facility. 403.19 Section 403.19 Protection of Environment... Owatonna Waste Water Treatment Facility. (a) For the purposes of this section, the term “Participating... Industrial User discharging to the Owatonna Waste Water Treatment Facility in Owatonna, Minnesota, when a...

40 CFR 122.24 - Concentrated aquatic animal production facilities (applicable to State NPDES programs, see § 123...

Code of Federal Regulations, 2010 CFR

2010-07-01

... Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS EPA ADMINISTERED PERMIT PROGRAMS: THE... animal production facility means a hatchery, fish farm, or other facility which meets the criteria in... any warm or cold water aquatic animal production facility as a concentrated aquatic animal production...

40 CFR 122.24 - Concentrated aquatic animal production facilities (applicable to State NPDES programs, see § 123...

Code of Federal Regulations, 2011 CFR

2011-07-01

... Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS EPA ADMINISTERED PERMIT PROGRAMS: THE... animal production facility means a hatchery, fish farm, or other facility which meets the criteria in... any warm or cold water aquatic animal production facility as a concentrated aquatic animal production...

Weightless Environment Training Facility (WETF) materials coating evaluation, volume 3

NASA Technical Reports Server (NTRS)

1995-01-01

This volume consists of Appendices C, D, E, and F to the report on the Weightless Environment Training Facility Materials Coating Evaluation project. The project selected 10 coating systems to be evaluated in six separate exposure environments, and subject to three tests for physical properties. Appendix C is the photographic appendix of the test panels. Appendix D details methods and procedures. Appendix E lists application equipment costs. Appendix F is a compilation of the solicitation of the candidate coating systems.

Design considerations for combined radiation effects facilities for twelve year outer planet spacecraft voyages

NASA Technical Reports Server (NTRS)

Miller, C. G.

1972-01-01

The design considerations influencing the choice and utility of environmental simulation methods and facilities are described, insofar as they relate to the requirements imposed on outer planet spacecraft because of radiation environments to be expected. Possible means for duplicating the radioisotope thermoelectric generator radiation environment, and for duplicating the effects of the trapped radiation belt environment are described, together with an assessment of radiation levels to be expected in the vicinity of an environmental testing chamber when in use.

Weightless Environment Training Facility (WETF) Materials Coating Evaluation, Volume 1

NASA Technical Reports Server (NTRS)

1995-01-01

The Weightless Environment Training Facility Material Coating Evaluation project has included preparing, coating, testing, and evaluating 800 test panels of three differing substrates. Ten selected coating systems were evaluated in six separate exposure environments and subject to three tests for physical properties. Substrate materials were identified, the manner of surface preparation described, and exposure environments defined. Exposure environments included immersion exposure, cyclic exposure, and field exposure. Cyclic exposures, specifically QUV-Weatherometer and the KTA Envirotest were found to be the most agressive of the environments included in the study when all three evaluation criteria are considered. This was found to result primarily from chalking of the coatings under ultraviolet (UV) light exposure. Volumes 2 and 3 hold the 5 appendices to this report.

Environment, Safety, and Health: Status of DOE’s Reorganization of it’s Safety Oversight Function

DTIC Science & Technology

1990-01-01

facilities. After deliberation, the Congress in late 1988 directed that the Defense Nuclear Facilities Safety Board be established to provide...nuclear safety matters will be conducted by either the Advisory Committee on Nuclear Facility Safety or the recently mandated Defense Nuclear Facilities Safety...the facilities under the statutory purview of the Defense Nuclear Facilities Safety Board once the board determines it is ready to assume independent

The food and beverage vending environment in health care facilities participating in the healthy eating, active communities program.

PubMed

Lawrence, Sally; Boyle, Maria; Craypo, Lisa; Samuels, Sarah

2009-06-01

Little has been done to ensure that the foods sold within health care facilities promote healthy lifestyles. Policies to improve school nutrition environments can serve as models for health care organizations. This study was designed to assess the healthfulness of foods sold in health care facility vending machines as well as how health care organizations are using policies to create healthy food environments. Food and beverage assessments were conducted in 19 California health care facilities that serve children in the Healthy Eating, Active Communities sites. Items sold in vending machines were inventoried at each facility and interviews conducted for information on vending policies. Analyses examined the types of products sold and the healthfulness of these products. Ninety-six vending machines were observed in 15 (79%) of the facilities. Hospitals averaged 9.3 vending machines per facility compared with 3 vending machines per health department and 1.4 per clinic. Sodas comprised the greatest percentage of all beverages offered for sale: 30% in hospital vending machines and 38% in clinic vending machines. Water (20%) was the most prevalent in health departments. Candy comprised the greatest percentage of all foods offered in vending machines: 31% in clinics, 24% in hospitals, and 20% in health department facilities. Across all facilities, 75% of beverages and 81% of foods sold in vending machines did not adhere to the California school nutrition standards (Senate Bill 12). Nine (47%) of the health care facilities had adopted, or were in the process of adopting, policies that set nutrition standards for vending machines. According to the California school nutrition standards, the majority of items found in the vending machines in participating health care facilities were unhealthy. Consumption of sweetened beverages and high-energy-density foods has been linked to increased prevalence of obesity. Some health care facilities are developing policies that set nutrition standards for vending machines. These policies could be effective in increasing access to healthy foods and beverages in institutional settings.

40 CFR 257.2 - Definitions.

Code of Federal Regulations, 2011 CFR

2011-07-01

... Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES CRITERIA FOR CLASSIFICATION OF SOLID WASTE DISPOSAL FACILITIES AND PRACTICES Classification of Solid Waste Disposal Facilities... demolition (C&D) landfill means a solid waste disposal facility subject to the requirements of subparts A or...

SSC OCIO, IT SUMMIT 2011

NASA Technical Reports Server (NTRS)

Cottrell, Dinna L.

2011-01-01

The Stennis Space Center (SSC) Records Retention Facility is a centralized location for all SSC records, Records Management staff, and the SSC History Office. The building is a storm resistant facility and provides a secure environment for records housing. The Records Retention Facility was constructed in accordance with The National Archives and Records Administration (NARA) requirements for records storage, making it the first NARA compliant facility in the agency. Stennis Space Center's Records Retention Facility became operational in May 2010. The SSC Records Retention Facility ensures that the required federal records are preserved, managed and accessible to all interested personnel. The facility provides 20,000 cubic feet of records storage capacity for the purpose of managing the centers consolidated records within a central, protected environment. Records housed in the facility are in the form of paper, optical, film and magnetic media. Located within the SSC Records Retention Facility, the Records Management Office provides comprehensive records management services in the form of: a) Storage and life-cycle management of inactive records of all media types; b) Digitizing/scanning of records and documents; c) Non-textual/digital electronic records media storage, migration and transfer; d) Records Remediation.

Evaluation of the built environment: staff and family satisfaction pre- and post-occupancy of the Children's Hospital.

PubMed

Kotzer, Anne Marie; Zacharakis, Susan Koch; Raynolds, Mary; Buenning, Fred

2011-01-01

To evaluate and compare the impact of an existing and newly built hospital environment on family and staff satisfaction related to light, noise, temperature, aesthetics, and amenities, as well as safety, security, and privacy. The United States is engaged in an unprecedented healthcare building boom driven by the need to replace aging facilities, understand the impact of the built environment on quality and safety, incorporate rapidly emerging technologies, and enhance patient- and family-centered care. More importantly, there is heightened attention to creating optimal physical environments to achieve the best possible outcomes for patients, families, and staff. Using a pre-post descriptive survey design, all nursing, social work, therapy staff, and families on selected inpatient units were invited to participate. A demographic form and Family and Staff Satisfaction Surveys were developed and administered pre- and post-occupancy of the new facility. Pre/post mean scores for staff satisfaction improved on all survey subscales with statistically significant improvement (p < .05) in most areas. The most improvement was seen with layout of the patient room, natural light, storage and writing surfaces, and comfort and appeal. Family satisfaction demonstrated statistically significant improvement on all subscales (p ≤ .01), especially for natural light, quiet space, parking, and the child's room as a healing environment. Families and staff reported greater satisfaction with the newly built hospital environment compared to the old facility. Study results will help guide future architectural design decisions, attract and retain staff at a world-class facility, and create the most effective healing environments.

40 CFR 280.220 - Ownership of an underground storage tank or underground storage tank system or facility or...

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 26 2010-07-01 2010-07-01 false Ownership of an underground storage tank or underground storage tank system or facility or property on which an underground storage tank or underground storage tank system is located. 280.220 Section 280.220 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID...

40 CFR Appendix B to Subpart I of... - Allowance for Facilities Planning and Design

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 1 2012-07-01 2012-07-01 false Allowance for Facilities Planning and Design B Appendix B to Subpart I of Part 35 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY GRANTS AND OTHER FEDERAL ASSISTANCE STATE AND LOCAL ASSISTANCE Grants for Construction of Treatment Works Pt. 35, Subpt. I, App. B Appendix B to...

A ``Cyber Wind Facility'' for HPC Wind Turbine Field Experiments

NASA Astrophysics Data System (ADS)

Brasseur, James; Paterson, Eric; Schmitz, Sven; Campbell, Robert; Vijayakumar, Ganesh; Lavely, Adam; Jayaraman, Balaji; Nandi, Tarak; Jha, Pankaj; Dunbar, Alex; Motta-Mena, Javier; Craven, Brent; Haupt, Sue

2013-03-01

The Penn State ``Cyber Wind Facility'' (CWF) is a high-fidelity multi-scale high performance computing (HPC) environment in which ``cyber field experiments'' are designed and ``cyber data'' collected from wind turbines operating within the atmospheric boundary layer (ABL) environment. Conceptually the ``facility'' is akin to a high-tech wind tunnel with controlled physical environment, but unlike a wind tunnel it replicates commercial-scale wind turbines operating in the field and forced by true atmospheric turbulence with controlled stability state. The CWF is created from state-of-the-art high-accuracy technology geometry and grid design and numerical methods, and with high-resolution simulation strategies that blend unsteady RANS near the surface with high fidelity large-eddy simulation (LES) in separated boundary layer, blade and rotor wake regions, embedded within high-resolution LES of the ABL. CWF experiments complement physical field facility experiments that can capture wider ranges of meteorological events, but with minimal control over the environment and with very small numbers of sensors at low spatial resolution. I shall report on the first CWF experiments aimed at dynamical interactions between ABL turbulence and space-time wind turbine loadings. Supported by DOE and NSF.

Vulnerability of CMOS image sensors in Megajoule Class Laser harsh environment.

PubMed

Goiffon, V; Girard, S; Chabane, A; Paillet, P; Magnan, P; Cervantes, P; Martin-Gonthier, P; Baggio, J; Estribeau, M; Bourgade, J-L; Darbon, S; Rousseau, A; Glebov, V Yu; Pien, G; Sangster, T C

2012-08-27

CMOS image sensors (CIS) are promising candidates as part of optical imagers for the plasma diagnostics devoted to the study of fusion by inertial confinement. However, the harsh radiative environment of Megajoule Class Lasers threatens the performances of these optical sensors. In this paper, the vulnerability of CIS to the transient and mixed pulsed radiation environment associated with such facilities is investigated during an experiment at the OMEGA facility at the Laboratory for Laser Energetics (LLE), Rochester, NY, USA. The transient and permanent effects of the 14 MeV neutron pulse on CIS are presented. The behavior of the tested CIS shows that active pixel sensors (APS) exhibit a better hardness to this harsh environment than a CCD. A first order extrapolation of the reported results to the higher level of radiation expected for Megajoule Class Laser facilities (Laser Megajoule in France or National Ignition Facility in the USA) shows that temporarily saturated pixels due to transient neutron-induced single event effects will be the major issue for the development of radiation-tolerant plasma diagnostic instruments whereas the permanent degradation of the CIS related to displacement damage or total ionizing dose effects could be reduced by applying well known mitigation techniques.

Medical education and social environment.

PubMed

Rasool, Ahsan; Qayum, Iftikhar; Ahmad, Ashfaq; Farooq, Umer; Shah, Awais Ali; Waqas, Muhammad; Rasool, Maleeha; Hameed, Sania; Kanwal, Rana; Azmat, Muneeba; Marwat, Saleem; Afridit, Faheem

2014-01-01

A positive learning environment and quality of course content have an imperative role in academic achievement of students. This study aims to assess students' point of view about the quality of education and social environment of a public sector medical college in Pakistan. Relative scarcity of data from students' perspective merited this study. A cross-sectional survey was undertaken at Ayub Medical College, Abbottabad, Pakistan, including 300 medical students from all five years of the MBBS course. Systematic random sampling was used with a kth interval of 4 for each class. Self-administered questionnaire was used and contained items related to academics, learning environment, learning resources, teaching methodologies and student-friendly activities. The data were analysed using SPSS-16. There were 265 respondents (88.3%) to the questionnaire with males accounting for 58.9% (n=156). In general students showed satisfaction with quality of content being taught; however there was discontent towards various academic and non- academic facilities provided to the students. Only 44.10% and 31.50% students reported provision of academic related facilities and interactive sessions as up to mark respectively; 83% students reported that undergraduate medical research was in need of improvement; 55.5% and 60.2% reported that facilities in hostel and recreational facilities needed improvement respectively; and 52.8% students stated presence of a healthy, student friendly, encouraging environment was not up to mark. Although course content and teaching methodologies are generally satisfactory, a healthy, student friendly, encouraging environment is vet to be created to help students foster their abilities completely.

40 CFR 60.30d - Designated facilities.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 6 2010-07-01 2010-07-01 false Designated facilities. 60.30d Section... Acid Production Units § 60.30d Designated facilities. Sulfuric acid production units. The designated facility to which §§ 60.31d and 60.32d apply is each existing “sulfuric acid production unit” as defined in...

15 CFR 971.606 - Onshore information.

Code of Federal Regulations, 2013 CFR

2013-01-01

... environment of port, transport, processing and waste disposal facilities and associated facilities (e.g., maps... to enable NOAA to function as lead agency in preparing permit site-specific environmental impact... construction and operation of the facilities, including waste characteristics and toxicity; (3) Any mitigating...

15 CFR 971.606 - Onshore information.

Code of Federal Regulations, 2014 CFR

2014-01-01

... environment of port, transport, processing and waste disposal facilities and associated facilities (e.g., maps... to enable NOAA to function as lead agency in preparing permit site-specific environmental impact... construction and operation of the facilities, including waste characteristics and toxicity; (3) Any mitigating...

Contamination control and cleanliness level integrity for the Space Shuttle Orbiter PLB, payloads and facilities at KSC

NASA Technical Reports Server (NTRS)

Bartelson, D.

1984-01-01

The PLB, its cargo, and payload canister must satisfy the cleanliness requirements of visual clean (VC) level 1, 2, 3, or special as stated in NASA document SN-C-0005A. The specific level of cleanliness is chosen by the payload bay customer for their mission. During orbiter turnaround processing at KSC, the payload bay is exposed to the environments of the Orbiter Processing Facility (OPF) and the Payload Changeout Room (PCR). In supportive response to the orbiter payload bay/facility interface, it is necessary that the facility environment be controlled and monitored to protect the cleanliness/environmental integrity of the payload bay and its cargo. Techniques used to meet environmental requirements during orbiter processing are introduced.

40 CFR 60.434 - Monitoring of operations and recordkeeping.

Code of Federal Regulations, 2011 CFR

2011-07-01

... recordkeeping. 60.434 Section 60.434 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR... affected facility using waterborne ink systems or solvent-borne ink systems with solvent recovery systems...) If affected facilities share the same raw ink storage/handling system with existing facilities...

40 CFR 60.434 - Monitoring of operations and recordkeeping.

Code of Federal Regulations, 2010 CFR

2010-07-01

... recordkeeping. 60.434 Section 60.434 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR... affected facility using waterborne ink systems or solvent-borne ink systems with solvent recovery systems...) If affected facilities share the same raw ink storage/handling system with existing facilities...

Assessment of air velocity sensors for use in animal produciton facilities

USDA-ARS?s Scientific Manuscript database

Ventilation is an integral part of thermal environment control in animal production facilities. Accurately measuring the air velocity distribution within these facilities is cumbersome using the traverse method and a distributed velocity measurement system would reduce the time necessary to perform ...

Conceptual Design of Tail-Research EXperiment (T-REX) on Space Plasma Environment Research Facility

NASA Astrophysics Data System (ADS)

Xiao, Qingmei; Wang, Xiaogang; E, Peng; Shen, Chao; Wang, Zhibin; Mao, Aohua; Xiao, Chijie; Ding, Weixing; Ji, Hantao; Ren, Yang

2016-10-01

Space Environment Simulation Research Infrastructure (SESRI), a scientific project for a major national facility of fundamental researches, has recently been launched at Harbin Institute of Technology (HIT). The Space Plasma Environment Research Facility (SPERF) for simulation of space plasma environment is one of the components of SESRI. It is designed to investigate fundamental issues in space plasma environment, such as energetic particles transportation and the interaction with waves in magnetosphere, magnetic reconnection at magnetopause and magnetotail, etc. Tail-Research Experiment (T-REX) is part of the SPERF for laboratory studies of space physics relevant to tail reconnection and dipolarization process. T-REX is designed to carry out two kinds of experiments: the tail plasmamoid for magnetic reconnection and magnetohydrodynamic waves excited by high speed plasma jet. In this presentation, the scientific goals and experimental plans for T-REX together with the means applied to generate the plasma with desired parameters are reviewed. Two typical scenarios of T-REX with operations of plasma sources and various magnetic configurations to study specific physical processes in space plasmas will also be presented.

Hyperthermal Environments Simulator for Nuclear Rocket Engine Development

NASA Technical Reports Server (NTRS)

Litchford, Ron J.; Foote, John P.; Clifton, W. B.; Hickman, Robert R.; Wang, Ten-See; Dobson, Christopher C.

2011-01-01

An arc-heater driven hyperthermal convective environments simulator was recently developed and commissioned for long duration hot hydrogen exposure of nuclear thermal rocket materials. This newly established non-nuclear testing capability uses a high-power, multi-gas, wall-stabilized constricted arc-heater to produce hightemperature pressurized hydrogen flows representative of nuclear reactor core environments, excepting radiation effects, and is intended to serve as a low-cost facility for supporting non-nuclear developmental testing of hightemperature fissile fuels and structural materials. The resulting reactor environments simulator represents a valuable addition to the available inventory of non-nuclear test facilities and is uniquely capable of investigating and characterizing candidate fuel/structural materials, improving associated processing/fabrication techniques, and simulating reactor thermal hydraulics. This paper summarizes facility design and engineering development efforts and reports baseline operational characteristics as determined from a series of performance mapping and long duration capability demonstration tests. Potential follow-on developmental strategies are also suggested in view of the technical and policy challenges ahead. Keywords: Nuclear Rocket Engine, Reactor Environments, Non-Nuclear Testing, Fissile Fuel Development.

A Multidisciplinary Paradigm and Approach to Protecting Human Health and the Environment, Society, and Stakeholders at Nuclear Facilities - 12244

DOE Office of Scientific and Technical Information (OSTI.GOV)

Burger, Joanna; Environmental and Occupational Health Sciences Institute, Piscataway, NJ; Gochfeld, Michael

2012-07-01

As the Department of Energy (DOE) continues to remediate its lands, and to consider moving toward long-term stewardship and the development of energy parks on its industrial, remediated land, it is essential to adequately characterize the environment around such facilities to protect society, human health, and the environment. While DOE sites re considering several different land-use scenarios, all of them require adequate protection of the environment. Even if DOE lands are developed for energy parks that are mainly for industrializes sections of DOE lands that will not be remediated to residential standards, there is still the need to consider themore » protection of human health and the environment. We present an approach to characterization and establishment of teams that will gather the information, and integrate that information for a full range of stakeholders from technical personnel, to public policy makers, and that public. Such information is needed to establish baselines, site new energy facilities in energy parks, protect existing nuclear facilities and nuclear wastes, improve the basis for emergency planning, devise suitable monitoring schemes to ensure continued protection, provide data to track local and regional response changes, and for mitigation, remediation and decommissioning planning. We suggest that there are five categories of information or data needs, including 1) geophysical, sources, fate and transport, 2) biological systems, 3) human health, 4) stakeholder and environmental justice, and 5) societal, economic, and political. These informational needs are more expansive than the traditional site characterization, but encompass a suite of physical, biological, and societal needs to protect all aspects of human health and the environment, not just physical health. We suggest a Site Committee be established that oversees technical teams for each of the major informational categories, with appropriate representation among teams and with a broad involvement of a range of governmental personnel, natural and social scientists, Native Americans, environmental justice communities, and other stakeholders. Such informational teams (and Oversight Committee) would report to a DOE-designated authority or Citizen's Advisory Board. Although designed for nuclear facilities and energy parks on DOE lands, the templates and information teams can be adapted for other hazardous facilities, such as a mercury storage facility at Oak Ridge. (authors)« less

Removal site evaluation report for the Isotope Facilities at Oak Ridge National Laboratory, Oak Ridge, Tennessee

DOE Office of Scientific and Technical Information (OSTI.GOV)

NONE

This removal site evaluation (RmSE) report of the Isotope Facilities at Oak Ridge National Laboratory (ORNL) was prepared to provide the Environmental Restoration Program with information necessary to evaluate whether hazardous and/or radiological contaminants in and around the Isotopes Facility pose a substantial risk to human health or the environment and if remedial site evaluations (RSEs) or removal actions are required. The scope of the project included: (1) a review of historical evidence regarding operations and use of the facility; (2) interviews with facility personnel concerning current and past operating practices; (3) a site inspection; and (4) identification of hazardmore » areas requiring maintenance, removal, or remedial actions. The results of RmSE indicate that no substantial risks exist from contaminants present in the Isotope Facilities because adequate controls and practices exist to protect human health and the environment. The recommended correction from the RmSE are being conducted as maintenance actions; accordingly, this RmSE is considered complete and terminated.« less

Quality assurance procedures for environmental control and monitoring in plant growth facilities. Report of the North Central Regional 101 Committee on Growth Chamber Use

NASA Technical Reports Server (NTRS)

Tibbitts, T. W. (Principal Investigator)

1986-01-01

This report includes procedures for ensuring the quality of the environment provided for plant growth in controlled environment facilities. Biologists and engineers may use these procedures for ensuring quality control during experiments or for ensuring quality control in the design of plant growth facilities. Environmental monitoring prior to and during experiments is included in these procedures. Specific recommendations cover control, acquisition, and calibration for sensor types for the separate parameters of radiation (light), temperature, humidity, carbon dioxide, and air movement.

THE LARGE HIGH PRESSURE ARC PLASMA GENERATOR: A FACILITY FOR SIMULATING MISSLE AND SATELLITE RE-ENTRY. Research Report 56

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rose, P.; Powers, W.; Hritzay, D.

1959-06-01

The development of an arc wind tunnel capable of stagnation pressures in the excess of twenty atmospheres and using as much as fifteen megawatts of electrical power is described. The calibration of this facility shows that it is capable of reproducing the aerodynamic environment encountered by vehicles flying at velocities as great as satellite velocity. Its use as a missile re-entry material test facility is described. The large power capacity of this facility allows one to make material tests on specimens of size sufficient to be useful for material development yet at realistic energy and Reynolds number values. By themore » addition of a high-capacity vacuum system, this facility can be used to produce the low density, high Mach number environment needed for simulating satellite re-entry, as well as hypersonic flight at extreme altitudes. (auth)« less

Chemical Facility Security: Reauthorization, Policy Issues, and Options for Congress

DTIC Science & Technology

2010-11-15

American Institute of Chemical Engineers , before the Senate Committee on Environment and Public Works, June 21, 2006, S.Hrg. 109-1044. See also...American Institute of Chemical Engineers , before the Senate Committee on Environment and Public Works, June 21, 2006, S.Hrg. 109-1044. 57 The DHS...CRS Report for Congress Prepared for Members and Committees of Congress Chemical Facility Security: Reauthorization, Policy Issues, and

40 CFR 355.20 - If this subpart applies to my facility, what information must I provide, who must I submit it to...

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 28 2011-07-01 2011-07-01 false If this subpart applies to my facility, what information must I provide, who must I submit it to, and when is it due? 355.20 Section 355.20 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SUPERFUND, EMERGENCY PLANNING, AND...