The WRKY transcription factor family and senescence in switchgrass.

PubMed

Rinerson, Charles I; Scully, Erin D; Palmer, Nathan A; Donze-Reiner, Teresa; Rabara, Roel C; Tripathi, Prateek; Shen, Qingxi J; Sattler, Scott E; Rohila, Jai S; Sarath, Gautam; Rushton, Paul J

2015-11-09

Early aerial senescence in switchgrass (Panicum virgatum) can significantly limit biomass yields. WRKY transcription factors that can regulate senescence could be used to reprogram senescence and enhance biomass yields. All potential WRKY genes present in the version 1.0 of the switchgrass genome were identified and curated using manual and bioinformatic methods. Expression profiles of WRKY genes in switchgrass flag leaf RNA-Seq datasets were analyzed using clustering and network analyses tools to identify both WRKY and WRKY-associated gene co-expression networks during leaf development and senescence onset. We identified 240 switchgrass WRKY genes including members of the RW5 and RW6 families of resistance proteins. Weighted gene co-expression network analysis of the flag leaf transcriptomes across development readily separated clusters of co-expressed genes into thirteen modules. A visualization highlighted separation of modules associated with the early and senescence-onset phases of flag leaf growth. The senescence-associated module contained 3000 genes including 23 WRKYs. Putative promoter regions of senescence-associated WRKY genes contained several cis-element-like sequences suggestive of responsiveness to both senescence and stress signaling pathways. A phylogenetic comparison of senescence-associated WRKY genes from switchgrass flag leaf with senescence-associated WRKY genes from other plants revealed notable hotspots in Group I, IIb, and IIe of the phylogenetic tree. We have identified and named 240 WRKY genes in the switchgrass genome. Twenty three of these genes show elevated mRNA levels during the onset of flag leaf senescence. Eleven of the WRKY genes were found in hotspots of related senescence-associated genes from multiple species and thus represent promising targets for future switchgrass genetic improvement. Overall, individual WRKY gene expression profiles could be readily linked to developmental stages of flag leaves.

PAF53 is essential in mammalian cells: CRISPR/Cas9 fails to eliminate PAF53 expression.

PubMed

Rothblum, Lawrence I; Rothblum, Katrina; Chang, Eugenie

2017-05-15

When mammalian cells are nutrient and/or growth factor deprived, exposed to inhibitors of protein synthesis, stressed by heat shock or grown to confluence, rDNA transcription is essentially shut off. Various mechanisms are available to accomplish this downshift in ribosome biogenesis. Muramatsu's laboratory (Hanada et al., 1996) first demonstrated that mammalian PAF53 was essential for specific rDNA transcription and that PAF53 levels were regulated in response to growth factors. While S. cerevisae A49, the homologue of vertebrate PAF53, is not essential for viability (Liljelund et al., 1992), deletion of yA49 results in colonies that grow at 6% of the wild type rate at 25°C. Experiments described by Wang et al. (2015) identified PAF53 as a gene "essential for optimal proliferation". However, they did not discriminate genes essential for viability. Hence, in order to resolve this question, we designed a series of experiments to determine if PAF53 was essential for cell survival. We set out to delete the gene product from mammalian cells using CRISPR/CAS9 technology. Human 293 cells were transfected with lentiCRISPR v2 carrying genes for various sgRNA that targeted PAF53. In some experiments, the cells were cotransfected in parallel with plasmids encoding FLAG-tagged mouse PAF53. After treating the transfected cells with puromycin (to select for the lentiCRISPR backbone), cells were cloned and analyzed by western blots for PAF53 expression. Genomic DNA was amplified across the "CRISPRd" exon, cloned and sequenced to identify mutated PAF53 genes. We obtained cell lines in which the endogenous PAF53 gene was "knocked out" only when we rescued with FLAG-PAF53. DNA sequencing demonstrated that in the absence of ectopic PAF53 expression, cells demonstrated unique means of surviving; including recombination or the utilization of alternative reading frames. We never observed a clone in which one PAF53 gene is expressed, unless there was also ectopic expression In the absence of ectopic gene expression, the gene products of both endogenous genes were expressed, irrespective of whether they were partially mutant proteins or not. Copyright © 2016 Elsevier B.V. All rights reserved.

Chicken TREM-B1, an Inhibitory Ig-Like Receptor Expressed on Chicken Thrombocytes

PubMed Central

Turowski, Vanessa; Sperling, Beatrice; Hanczaruk, Matthias A.; Göbel, Thomas W.; Viertlboeck, Birgit C.

2016-01-01

Triggering receptors expressed on myeloid cells (TREM) form a multigene family of immunoregulatory Ig-like receptors and play important roles in the regulation of innate and adaptive immunity. In chickens, three members of the TREM family have been identified on chromosome 26. One of them is TREM-B1 which possesses two V-set Ig-domains, an uncharged transmembrane region and a long cytoplasmic tail with one ITSM and two ITIMs indicating an inhibitory function. We generated specific monoclonal antibodies by immunizing a Balb/c mouse with a TREM-B1-FLAG transfected BWZ.36 cell line and tested the hybridoma supernatants on TREM-B1-FLAG transfected 2D8 cells. We obtained two different antibodies specific for TREM-B1, mab 7E8 (mouse IgG1) and mab 1E9 (mouse IgG2a) which were used for cell surface staining. Single and double staining of different tissues, including whole blood preparations, revealed expression on thrombocytes. Next we investigated the biochemical properties of TREM-B1 by using the specific mab 1E9 for immunoprecipitation of either lysates of surface biotinylated peripheral blood cells or stably transfected 2D8 cells. Staining with streptavidin coupled horse radish peroxidase revealed a glycosylated monomeric protein of about 50 kDa. Furthermore we used the stably transfected 2D8 cell line for analyzing the cytoplasmic tyrosine based signaling motifs. After pervanadate treatment, we detected phosphorylation of the tyrosine residues and subsequent recruitment of the tyrosine specific protein phosphatase SHP-2, indicating an inhibitory potential for TREM-B1. We also showed the inhibitory effect of TREM-B1 in chicken thrombocytes using a CD107 degranulation assay. Crosslinking of TREM-B1 on activated primary thrombocytes resulted in decreased CD107 surface expression of about 50–70%. PMID:26967520

Customization of Advia 120 thresholds for canine erythrocyte volume and hemoglobin concentration, and effects on morphology flagging results.

PubMed

Grimes, Carolyn N; Fry, Michael M

2014-12-01

This study sought to develop customized morphology flagging thresholds for canine erythrocyte volume and hemoglobin concentration [Hgb] on the ADVIA 120 hematology analyzer; compare automated morphology flagging with results of microscopic blood smear evaluation; and examine effects of customized thresholds on morphology flagging results. Customized thresholds were determined using data from 52 clinically healthy dogs. Blood smear evaluation and automated morphology flagging results were correlated with mean cell volume (MCV) and cellular hemoglobin concentration mean (CHCM) in 26 dogs. Customized thresholds were applied retroactively to complete blood (cell) count (CBC) data from 5 groups of dogs, including a reference sample group, clinical cases, and animals with experimentally induced iron deficiency anemia. Automated morphology flagging correlated more highly with MCV or CHCM than did blood smear evaluation; correlation with MCV was highest using customized thresholds. Customized morphology flagging thresholds resulted in more sensitive detection of microcytosis, macrocytosis, and hypochromasia than default thresholds.

Children's Interpretation of Focus Expressions in English and Mandarin

ERIC Educational Resources Information Center

Notley, Anna; Zhou, Peng; Crain, Stephen; Thornton, Rosalind

2009-01-01

Children often produce nonadult responses to sentences with the focus operator only, such as "Only the cat is holding a flag." For example, children often accept this sentence as a description of a situation in which a cat holds a flag and a duck holds both a flag and a balloon. One proposed analysis, by Paterson, Liversedge, Rowland & Filik…

Purification of FLAG-tagged Secreted Proteins from Mammalian Cells

PubMed Central

Itakura, Eisuke; Chen, Changchun; de Bono, Mario

2017-01-01

This protocol describes a method for purifying glycosylated FLAG-tagged secreted proteins with disulfide bonds from mammalian cells. The purified products can be used for various applications, such as ligand binding assays. PMID:29075655

Novel Function of NIBP in Breast Cancer

DTIC Science & Technology

2012-05-01

reduced in NIBP knockdown cells (Fig. 8). 7 pRK -Flag-NIBP Isoforms(aa) 960 944 1200 1246 1148 S E A P A c ti v it y ( F o ld...Fig.9. MDA-MB-231 cells were co-transfected by TurboFectin8.0 with empty pRK -Flag vector or various isoforms of NIBP with NF-B-SEAP reporter and...Ser536) pRK -Flag 12060301550 NIBP-mutA 12060301550TNFα (min) Fig.11. MDA-MB-231 cells at 60% confluence in 6-well plates were transfected with empty

Expression and characterization of human CB1 cannabinoid receptor in methylotrophic yeast Pichia pastoris.

PubMed

Kim, Tae-Kang; Zhang, Rundong; Feng, Wenke; Cai, Jian; Pierce, William; Song, Zhao-Hui

2005-03-01

For the purpose of purification and structural characterization, the CB1 cannabinoid receptors are expressed in methylotrophic yeast Pichia pastoris. The expression plasmid was constructed in which the CB1 gene is under the control of the highly inducible promoter of P. pastoris alcohol oxidase I gene. To facilitate easy detection and purification, a FLAG tag was introduced at the N-terminal, a c-myc epitope and a hexahistidine tag were introduced at the C-terminal of the CB1. In membrane preparations of CB1 gene transformed yeast cells, Western blot analysis detected the expression of CB1 proteins. Radioligand binding assays demonstrated that the tagged CB1 receptors expressed in P. pastoris have a pharmacological profile similar to that of the untagged CB1 receptors expressed in mammalian systems. Furthermore, the tagged CB1 receptors were purified by anti-FLAG M2 affinity chromatography and the identity of the purified CB1 receptor proteins was confirmed by Western blot analysis. MALDI/TOF mass spectrometry analysis of the peptides extracted from tryptic digestions of purified CB1 preparations detected 17 peptide fragments derived from the CB1, thus further confirming the identity of the purified receptor. In conclusion, these data demonstrated for the first time that epitope tagged, functional CB1 cannabinoid receptors can be expressed in P. pastoris for purification and mass spectrometry characterization.

Assessment of advanced warning signs for flagging operations.

DOT National Transportation Integrated Search

1999-05-01

The Virginia Department of Transportation (VDOT) and several other state departments : of transportation have expressed interest in modifying the advanced warning sign for work zone : flagging operations. The advanced warning sign is intended to aler...

Field Evidence of Social Influence in the Expression of Political Preferences: The Case of Secessionists Flags in Barcelona

PubMed Central

Parravano, Antonio; Noguera, José A.; Hermida, Paula; Tena-Sánchez, Jordi

2015-01-01

Models of social influence have explored the dynamics of social contagion, imitation, and diffusion of different types of traits, opinions, and conducts. However, few behavioral data indicating social influence dynamics have been obtained from direct observation in “natural” social contexts. The present research provides that kind of evidence in the case of the public expression of political preferences in the city of Barcelona, where thousands of citizens supporting the secession of Catalonia from Spain have placed a Catalan flag in their balconies and windows. Here we present two different studies. 1) During July 2013 we registered the number of flags in 26% of the electoral districts in the city of Barcelona. We find that there is a large dispersion in the density of flags in districts with similar density of pro-independence voters. However, by comparing the moving average to the global mean we find that the density of flags tends to be fostered in electoral districts where there is a clear majority of pro-independence vote, while it is inhibited in the opposite cases. We also show that the distribution of flags in the observed districts deviates significantly from that of an equivalent random distribution. 2) During 17 days around Catalonia’s 2013 national holiday we observed the position at balcony resolution of the flags displayed in the facades of a sub-sample of 82 blocks. We compare the ‘clustering index’ of flags on the facades observed each day to thousands of equivalent random distributions. Again we provide evidence that successive hangings of flags are not independent events but that a local influence mechanism is favoring their clustering. We also find that except for the national holiday day the density of flags tends to be fostered in facades located in electoral districts where there is a clear majority of pro-independence vote. PMID:25961562

Expression and Stability of Foreign Epitopes Introduced into 3A Nonstructural Protein of Foot-and-Mouth Disease Virus

PubMed Central

Li, Pinghua; Bai, Xingwen; Cao, Yimei; Han, Chenghao; Lu, Zengjun; Sun, Pu; Yin, Hong; Liu, Zaixin

2012-01-01

Foot-and-mouth disease virus (FMDV) is an aphthovirus that belongs to the Picornaviridae family and causes one of the most important animal diseases worldwide. The capacity of other picornaviruses to express foreign antigens has been extensively reported, however, little is known about FMDV. To explore the potential of FMDV as a viral vector, an 11-amino-acid (aa) HSV epitope and an 8 aa FLAG epitope were introduced into the C-terminal different regions of 3A protein of FMDV full-length infectious cDNA clone. Recombinant viruses expressing the HSV or FLAG epitope were successfully rescued after transfection of both modified constructs. Immunofluorescence assay, Western blot and sequence analysis showed that the recombinant viruses stably maintained the foreign epitopes even after 11 serial passages in BHK-21 cells. The 3A-tagged viruses shared similar plaque phenotypes and replication kinetics to those of the parental virus. In addition, mice experimentally infected with the epitope-tagged viruses could induce tag-specific antibodies. Our results demonstrate that FMDV can be used effectively as a viral vector for the delivery of foreign tags. PMID:22848509

Mangiferin inhibits cell migration and invasion through Rac1/WAVE2 signalling in breast cancer.

PubMed

Deng, Qing; Tian, Yan-Xiao; Liang, JianJun

2018-04-01

Breast tumour progression results from the advancement of the disease to a metastatic phenotype. Rac1 and Cdc42 belong to the Rho family of genes that, together with their downstream effectors, Wiskott-Aldrich Syndrome protein-family verprolin-homologous protein 2 (WAVE2) and Arp2/3, assume a vital part in cytoskeletal rearrangement and the arrangement of film projections that advance malignant cell relocation and invasion. Mangiferin is a characteristic polyphenolic compound from Mangifera indica L. (Anacardiaceae), ordinarily referred to as mango, that is consumed worldwide as a natural product, including culinary and seasoning applications. Mangiferin delays breast malignancy development and progression by inhibiting different signalling pathways required in mitogenic signalling and metastatic progression. Studies were performed to analyse the impact of mangiferin on Rac1/WAVE2 flagging, relocation and invasion in highly metastatic human MDA-MB-231 mammary cells. Additional studies led to the observation that comparative treatment with mangiferin caused marked reduction in tumour cell movement and invasion. Taken together, these discoveries demonstrate that mangiferin treatment adequately hinders Rac1/WAVE2 flagging and diminishes metastatic phenotypic expression in malignant mammary cells, indicating that mangiferin may provide a benefit as a novel restorative approach in the treatment of metastatic breast cancer.

Glomerular Epithelial Cells-Targeted Heme Oxygenase-1 Over Expression in the Rat: Attenuation of Proteinuria in Secondary But Not Primary Injury.

PubMed

Atsaves, Vassilios; Makri, Panagiota; Detsika, Maria G; Tsirogianni, Alexandra; Lianos, Elias A

2016-01-01

Induction of heme oxygenase 1 (HO-1) in glomerular epithelial cells (GEC) in response to injury is poor and this may be a disadvantage. We, therefore, explored whether HO-1 overexpression in GEC can reduce proteinuria induced by puromycin aminonucleoside (PAN) or in anti-glomerular basement membrane (GBM) antibody (Ab)-mediated glomerulonephritis (GN). HO-1 overexpression in GEC (GECHO-1) of Sprague-Dawley rats was achieved by targeting a FLAG-human (h) HO-1 using transposon-mediated transgenesis. Direct GEC injury was induced by a single injection of PAN. GN was induced by administration of an anti-rat GBM Ab and macrophage infiltration in glomeruli was assessed by immunohistochemistry and western blot analysis, which was also used to assess glomerular nephrin expression. In GECHO-1 rats, FLAG-hHO-1 transprotein was co-immunolocalized with nephrin. Baseline glomerular HO-1 protein levels were higher in GECHO-1 compared to wild type (WT) rats. Administration of either PAN or anti-GBM Ab to WT rats increased glomerular HO-1 levels. Nephrin expression markedly decreased in glomeruli of WT or GECHO-1 rats treated with PAN. In anti-GBM Ab-treated WT rats, nephrin expression also decreased. In contrast, it was preserved in anti-GBM Ab-treated GECHO-1 rats. In these, macrophage infiltration in glomeruli and the ratio of urine albumin to urine creatinine (Ualb/Ucreat) were markedly reduced. There was no difference in Ualb/Ucreat between WT and GECHO-1 rats treated with PAN. Depending on the type of injury, HO-1 overexpression in GEC may or may not reduce proteinuria. Reduced macrophage infiltration and preservation of nephrin expression are putative mechanisms underlying the protective effect of HO-1 overexpression following immune injury. © 2016 S. Karger AG, Basel.

Reference genes for gene expression studies in wheat flag leaves grown under different farming conditions

PubMed Central

2011-01-01

Background Internal control genes with highly uniform expression throughout the experimental conditions are required for accurate gene expression analysis as no universal reference genes exists. In this study, the expression stability of 24 candidate genes from Triticum aestivum cv. Cubus flag leaves grown under organic and conventional farming systems was evaluated in two locations in order to select suitable genes that can be used for normalization of real-time quantitative reverse-transcription PCR (RT-qPCR) reactions. The genes were selected among the most common used reference genes as well as genes encoding proteins involved in several metabolic pathways. Findings Individual genes displayed different expression rates across all samples assayed. Applying geNorm, a set of three potential reference genes were suitable for normalization of RT-qPCR reactions in winter wheat flag leaves cv. Cubus: TaFNRII (ferredoxin-NADP(H) oxidoreductase; AJ457980.1), ACT2 (actin 2; TC234027), and rrn26 (a putative homologue to RNA 26S gene; AL827977.1). In addition of these three genes that were also top-ranked by NormFinder, two extra genes: CYP18-2 (Cyclophilin A, AY456122.1) and TaWIN1 (14-3-3 like protein, AB042193) were most consistently stably expressed. Furthermore, we showed that TaFNRII, ACT2, and CYP18-2 are suitable for gene expression normalization in other two winter wheat varieties (Tommi and Centenaire) grown under three treatments (organic, conventional and no nitrogen) and a different environment than the one tested with cv. Cubus. Conclusions This study provides a new set of reference genes which should improve the accuracy of gene expression analyses when using wheat flag leaves as those related to the improvement of nitrogen use efficiency for cereal production. PMID:21951810

[Identification of the interacting proteins with S100A8 or S100A9 by affinity purification and mass spectrometry].

PubMed

Wang, Jing; Zhang, Xuemei; Li, Zheng; Li, Xiayu; Ma, Jian; Shen, Shourong

2017-04-28

To identify the interacting proteins with S100A8 or S100A9 in HEK293 cell line by flag-tag affinity purification and liquid chromatography mass spectrometry/mass spectrometry (LC-MS/MS).
 Methods: The p3×Flag-CMV-S100A8 and p3×Flag-CMV-S100A9 expression vectors were constructed by inserting S100A8 or S100A9 coding sequence. The recombinant plasmids were then transfected into HEK293 cells. Affinity purification and LC-MS/MS were applied to identify the proteins interacting with S100A8 or S100A9. Bioinformatics analysis was used to seek the gene ontology of the interacting proteins. Co-immunoprecipitation (Co-IP) was applied to confirm the proteins interacted with S100A8 or S100A9.
 Results: Fourteen proteins including pyruvate kinase, muscle (PKM), nucleophosmin (NPM1) and eukaryotic translation initiation factor 5A (EIF5A), which potentially interacted with S100A8, were successfully identified by Flag-tag affinity purification followed by LC-MS/MS analysis. Six proteins, such as tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein epsilon (14-3-3ε) and PKM, which potentially interacted with S100A9, were successfully identified. Gene ontology analysis of the identified proteins suggested that proteins interacted with S100A8 or S100A9 were involved in several biological pathways, including canonical glycolysis, positive regulation of NF-κB transcription factor activity, negative regulation of apoptotic process, cell-cell adhesion, etc. Co-IP experiment confirmed that PKM2 can interact with both S100A8 and S100A9, and 14-3-3ε can interact with S100A8.
 Conclusion: PKM2 is identified to interact with both S100A8 and S100A9, while 14-3-3ε can interact with S100A9. These results may provide a new clue for the role of S100A8 or S100A9 in the progression of colitis-associated colorectal cancer.

Characterization and Imaging of Antibody-Coated Gold Nanoparticles for Targeted Treatment of Microbial Keratitis

NASA Astrophysics Data System (ADS)

Mahan, Matthew

Microbial keratitis (MK) is an infection of the cornea by pathogenic organisms that causes inflammation and irritation. It can lead to full or partial blindness if left untreated. Current clinical treatment methods rely on high frequency application of topical drugs which are subject to the issues of patient compliance and microbial resistance. In this work, gold nanoparticles (AuNP) were proposed as an alternative treatment method in light-based therapies. Particle formulation methods were investigated and assessed using transmission electron microscopy (TEM) and ultraviolet/visible spectroscopy (UV-Vis). AuNP of 20 nm diameter were used as platforms to attach monoclonal antibodies anti-FLAG or anti-F1 to enhance their cell-targeting ability as well as polyethylene glycol to reduce non-specific binding and protein adsorption. These functionalized particles were qualitatively assessed using UV-Vis. The antibody-functionalized AuNP were then assessed for their ability to attach directly to Pseudomonas aeruginosa, expressing FLAG peptide, or Aspergillus fumigatus, expressing the F1 receptor. Attachment was imaged using dark field microscopy, transmission electron microscopy, and fluorescence microscopy.

Evidence that the rabbit proton-peptide co-transporter PepT1 is a multimer when expressed in Xenopus laevis oocytes.

PubMed

Panitsas, Konstantinos-E; Boyd, C A R; Meredith, David

2006-04-01

To test whether the rabbit proton-coupled peptide transporter PepT1 is a multimer, we have employed a combination of transport assays, luminometry and site-directed mutagenesis. A functional epitope-tagged PepT1 construct (PepT1-FLAG) was co-expressed in Xenopus laevis oocytes with a non-functional but normally trafficked mutant form of the same transporter (W294F-PepT1). The amount of PepT1-FLAG cRNA injected into the oocytes was kept constant, while the amount of W294F-PepT1 cRNA was increased over the mole fraction range of 0 to 1. The uptake of [(3)H]-D: -Phe-L: -Gln into the oocytes was measured at pH(out) 5.5, and the surface expression of PepT1-FLAG was quantified by luminometry. As the mole fraction of injected W294F-PepT1 increased, the uptake of D: -Phe-L: -Gln decreased. This occurred despite the surface expression of PepT1-FLAG remaining constant, and so we can conclude that PepT1 must be a multimer. Assuming that PepT1 acts as a homomultimer, the best fit for the modelling suggests that PepT1 could be a tetramer, with a minimum requirement of two functional subunits in each protein complex. Western blotting also showed the presence of higher-order complexes of PepT1-FLAG in oocyte membranes. It should be noted that we cannot formally exclude the possibility that PepT1 interacts with unidentified Xenopus protein(s). The finding that PepT1 is a multimer has important implications for the molecular modelling of this protein.

Consumer preferences for front-of-pack calories labelling.

PubMed

van Kleef, Ellen; van Trijp, Hans; Paeps, Frederic; Fernández-Celemín, Laura

2008-02-01

In light of the emerging obesity pandemic, front-of-pack calories labels may be an important tool to assist consumers in making informed healthier food choices. However, there is little prior research to guide key decisions on whether caloric content should be expressed in absolute terms or relative to recommended daily intake, whether it should be expressed in per serving or per 100 g and whether the information should be further brought alive for consumers in terms of what the extra calorie intake implies in relation to activity levels. The present study aimed at providing more insight into consumers' appreciation of front-of-pack labelling of caloric content of food products and their specific preferences for alternative execution formats for such information in Europe. For this purpose, eight executions of front-of-pack calorie flags were designed and their appeal and information value were extensively discussed with consumers through qualitative research in four different countries (Germany, The Netherlands, France and the UK). The results show that calories are well-understood and that participants were generally positive about front-of-pack flags, particularly when flags are uniform across products. The most liked flags are the simpler flags depicting only the number of calories per serving or per 100 g, while more complex flags including references to daily needs or exercise and the flag including a phrase referring to balanced lifestyle were least preferred. Some relevant differences between countries were observed. Although participants seem to be familiar with the notion of calories, they do not seem to fully understand how to apply them. From the results, managerial implications for the design and implementation of front-of-pack calorie labelling as well as important directions for future research are discussed.

Consumer preferences for front-of-pack calories labelling

PubMed Central

van Kleef, Ellen; van Trijp, Hans; Paeps, Frederic; Fernández-Celemín, Laura

2008-01-01

Objective In light of the emerging obesity pandemic, front-of-pack calories labels may be an important tool to assist consumers in making informed healthier food choices. However, there is little prior research to guide key decisions on whether caloric content should be expressed in absolute terms or relative to recommended daily intake, whether it should be expressed in per serving or per 100 g and whether the information should be further brought alive for consumers in terms of what the extra calorie intake implies in relation to activity levels. The present study aimed at providing more insight into consumers’ appreciation of front-of-pack labelling of caloric content of food products and their specific preferences for alternative execution formats for such information in Europe. Design For this purpose, eight executions of front-of-pack calorie flags were designed and their appeal and information value were extensively discussed with consumers through qualitative research in four different countries (Germany, The Netherlands, France and the UK). Results The results show that calories are well-understood and that participants were generally positive about front-of-pack flags, particularly when flags are uniform across products. The most liked flags are the simpler flags depicting only the number of calories per serving or per 100 g, while more complex flags including references to daily needs or exercise and the flag including a phrase referring to balanced lifestyle were least preferred. Some relevant differences between countries were observed. Although participants seem to be familiar with the notion of calories, they do not seem to fully understand how to apply them. Conclusion From the results, managerial implications for the design and implementation of front-of-pack calorie labelling as well as important directions for future research are discussed. PMID:17601362

Increased androgenic sensitivity in the hind limb muscular system marks the evolution of a derived gestural display.

PubMed

Mangiamele, Lisa A; Fuxjager, Matthew J; Schuppe, Eric R; Taylor, Rebecca S; Hödl, Walter; Preininger, Doris

2016-05-17

Physical gestures are prominent features of many species' multimodal displays, yet how evolution incorporates body and leg movements into animal signaling repertoires is unclear. Androgenic hormones modulate the production of reproductive signals and sexual motor skills in many vertebrates; therefore, one possibility is that selection for physical signals drives the evolution of androgenic sensitivity in select neuromotor pathways. We examined this issue in the Bornean rock frog (Staurois parvus, family: Ranidae). Males court females and compete with rivals by performing both vocalizations and hind limb gestural signals, called "foot flags." Foot flagging is a derived display that emerged in the ranids after vocal signaling. Here, we show that administration of testosterone (T) increases foot flagging behavior under seminatural conditions. Moreover, using quantitative PCR, we also find that adult male S. parvus maintain a unique androgenic phenotype, in which androgen receptor (AR) in the hind limb musculature is expressed at levels ∼10× greater than in two other anuran species, which do not produce foot flags (Rana pipiens and Xenopus laevis). Finally, because males of all three of these species solicit mates with calls, we accordingly detect no differences in AR expression in the vocal apparatus (larynx) among taxa. The results show that foot flagging is an androgen-dependent gestural signal, and its emergence is associated with increased androgenic sensitivity within the hind limb musculature. Selection for this novel gestural signal may therefore drive the evolution of increased AR expression in key muscles that control signal production to support adaptive motor performance.

Increased androgenic sensitivity in the hind limb muscular system marks the evolution of a derived gestural display

PubMed Central

Mangiamele, Lisa A.; Fuxjager, Matthew J.; Schuppe, Eric R.; Taylor, Rebecca S.; Hödl, Walter; Preininger, Doris

2016-01-01

Physical gestures are prominent features of many species’ multimodal displays, yet how evolution incorporates body and leg movements into animal signaling repertoires is unclear. Androgenic hormones modulate the production of reproductive signals and sexual motor skills in many vertebrates; therefore, one possibility is that selection for physical signals drives the evolution of androgenic sensitivity in select neuromotor pathways. We examined this issue in the Bornean rock frog (Staurois parvus, family: Ranidae). Males court females and compete with rivals by performing both vocalizations and hind limb gestural signals, called “foot flags.” Foot flagging is a derived display that emerged in the ranids after vocal signaling. Here, we show that administration of testosterone (T) increases foot flagging behavior under seminatural conditions. Moreover, using quantitative PCR, we also find that adult male S. parvus maintain a unique androgenic phenotype, in which androgen receptor (AR) in the hind limb musculature is expressed at levels ∼10× greater than in two other anuran species, which do not produce foot flags (Rana pipiens and Xenopus laevis). Finally, because males of all three of these species solicit mates with calls, we accordingly detect no differences in AR expression in the vocal apparatus (larynx) among taxa. The results show that foot flagging is an androgen-dependent gestural signal, and its emergence is associated with increased androgenic sensitivity within the hind limb musculature. Selection for this novel gestural signal may therefore drive the evolution of increased AR expression in key muscles that control signal production to support adaptive motor performance. PMID:27143723

THRAP3 interacts with and inhibits the transcriptional activity of SOX9 during chondrogenesis.

PubMed

Sono, Takashi; Akiyama, Haruhiko; Miura, Shigenori; Deng, Jian Min; Shukunami, Chisa; Hiraki, Yuji; Tsushima, Yu; Azuma, Yoshiaki; Behringer, Richard R; Matsuda, Shuichi

2018-07-01

Sex-determining region Y (Sry)-box (Sox)9 is required for chondrogenesis as a transcriptional activator of genes related to chondrocyte proliferation, differentiation, and cartilage-specific extracellular matrix. Although there have been studies investigating the Sox9-dependent transcriptional complexes, not all their components have been identified. In the present study, we demonstrated that thyroid hormone receptor-associated protein (THRAP)3 is a component of a SOX9 transcriptional complex by liquid chromatography mass spectrometric analysis of FLAG-tagged Sox9-binding proteins purified from FLAG-HA-tagged Sox9 knock-in mice. Thrap3 knockdown in ATDC5 chondrogenic cells increased the expression of Collagen type II alpha 1 chain (Col2a1) without affecting Sox9 expression. THRAP3 and SOX9 overexpression reduced Col2a1 levels to a greater degree than overexpression of SOX9 alone. The negative regulation of SOX9 transcriptional activity by THRAP3 was mediated by interaction between the proline-, glutamine-, and serine-rich domain of SOX9 and the innominate domain of THRAP3. These results indicate that THRAP3 negatively regulates SOX9 transcriptional activity as a cofactor of a SOX9 transcriptional complex during chondrogenesis.

IFN-ε Is Constitutively Expressed by Cells of the Reproductive Tract and Is Inefficiently Secreted by Fibroblasts and Cell Lines

PubMed Central

Hermant, Pascale; Francius, Cédric; Clotman, Frédéric; Michiels, Thomas

2013-01-01

Type-I interferons (IFNs) form a large family of cytokines that primarily act to control the early development of viral infections. Typical type-I IFN genes, such as those encoding IFN-α or IFN-β are upregulated by viral infection in many cell types. In contrast, the gene encoding IFN-ε was reported to be constitutively expressed by cells of the female reproductive tract and to contribute to the protection against vaginal infections with herpes simplex virus 2 and Chlamydia muridarum. Our data confirm the lack of induction of IFN-ε expression after viral infection and the constitutive expression of IFN-ε by cells of the female but also of the male reproductive organs. Interestingly, when expressed from transfected expression plasmids in 293T, HeLa or Neuro2A cells, the mouse and human IFN-ε precursors were inefficiently processed and secretion of IFN-ε was minimal. Analysis of chimeric constructs produced between IFN-ε and limitin (IFN-ζ) showed that both the signal peptide and the mature moiety of IFN-ε contribute to poor processing of the precursor. Immunofluorescent detection of FLAG-tagged IFN-ε in transfected cells suggested that IFN-ε and chimeric proteins were defective for progression through the secretory pathway. IFN-ε did not, however, act intracellularly and impart an antiviral state to producing cells. Given the constitutive expression of IFN-ε in specialized cells and the poor processing of IFN-ε precursor in fibroblasts and cell lines, we hypothesize that IFN-ε secretion may require a co-factor specifically expressed in cells of the reproductive organs, that might secure the system against aberrant release of this IFN. PMID:23951133

Dynamic and nucleolin-dependent localization of human cytomegalovirus UL84 to the periphery of viral replication compartments and nucleoli.

PubMed

Bender, Brian J; Coen, Donald M; Strang, Blair L

2014-10-01

Protein-protein and protein-nucleic acid interactions within subcellular compartments are required for viral genome replication. To understand the localization of the human cytomegalovirus viral replication factor UL84 relative to other proteins involved in viral DNA synthesis and to replicating viral DNA in infected cells, we created a recombinant virus expressing a FLAG-tagged version of UL84 (UL84FLAG) and used this virus in immunofluorescence assays. UL84FLAG localization differed at early and late times of infection, transitioning from diffuse distribution throughout the nucleus to exclusion from the interior of replication compartments, with some concentration at the periphery of replication compartments with newly labeled DNA and the viral DNA polymerase subunit UL44. Early in infection, UL84FLAG colocalized with the viral single-stranded DNA binding protein UL57, but colocalization became less prominent as infection progressed. A portion of UL84FLAG also colocalized with the host nucleolar protein nucleolin at the peripheries of both replication compartments and nucleoli. Small interfering RNA (siRNA)-mediated knockdown of nucleolin resulted in a dramatic elimination of UL84FLAG from replication compartments and other parts of the nucleus and its accumulation in the cytoplasm. Reciprocal coimmunoprecipitation of viral proteins from infected cell lysates revealed association of UL84, UL44, and nucleolin. These results indicate that UL84 localization during infection is dynamic, which is likely relevant to its functions, and suggest that its nuclear and subnuclear localization is highly dependent on direct or indirect interactions with nucleolin. Importance: The protein-protein interactions among viral and cellular proteins required for replication of the human cytomegalovirus (HCMV) DNA genome are poorly understood. We sought to understand how an enigmatic HCMV protein critical for virus replication, UL84, localizes relative to other viral and cellular proteins required for HCMV genome replication and replicating viral DNA. We found that UL84 localizes with viral proteins, viral DNA, and the cellular nucleolar protein nucleolin in the subnuclear replication compartments in which viral DNA replication occurs. Unexpectedly, we also found localization of UL84 with nucleolin in nucleoli and showed that the presence of nucleolin is involved in localization of UL84 to the nucleus. These results add to previous work showing the importance of nucleolin in replication compartment architecture and viral DNA synthesis and are relevant to understanding UL84 function. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Intracellular transport and stability of varicella-zoster virus glycoprotein K

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

Hall, Susan L.; Govero, Jennifer L.; Heineman, Thomas C.

2007-02-20

VZV gK, an essential glycoprotein that is conserved among the alphaherpesviruses, is believed to participate in membrane fusion and cytoplasmic virion morphogenesis based on analogy to its HSV-1 homolog. However, the production of VZV gK-specific antibodies has proven difficult presumably due to its highly hydrophobic nature and, therefore, VZV gK has received limited study. To overcome this obstacle, we inserted a FLAG epitope into gK near its amino terminus and produced VZV recombinants expressing epitope-tagged gK (VZV gK-F). These recombinants grew indistinguishably from native VZV, and FLAG-tagged gK could be readily detected in VZV gK-F-infected cells. FACS analysis established thatmore » gK is transported to the plasma membrane of infected cells, while indirect immunofluorescence demonstrated that gK accumulates predominately in the Golgi. Using VZV gK-F-infected cells we demonstrated that VZV gK, like several other herpesvirus glycoproteins, is efficiently endocytosed from the plasma membrane. However, pulse-labeling experiments revealed that the half-life of gK is considerably shorter than that of other VZV glycoproteins including gB, gE and gH. This finding suggests that gK may be required in lower abundance than other viral glycoproteins during virion morphogenesis or viral entry.« less

Repartitioning the National Community: Political Visibility and Voice for Undocumented Immigrants in the Spring 2006 Immigration Rights Marches

ERIC Educational Resources Information Center

Galindo, Rene

2010-01-01

The historic immigration rights marches of 2006 placed the plight of undocumented immigrants in the national spotlight. Competing interpretations of the marches focused in part on the waving of Mexican flags by marchers. While some English-language media critics saw the flags as expressing political disloyalty to the United States, the marchers…

The flagellar protein FLAG1/SMP1 is a candidate for Leishmania-sand fly interaction.

PubMed

Di-Blasi, Tatiana; Lobo, Amanda R; Nascimento, Luanda M; Córdova-Rojas, Jose L; Pestana, Karen; Marín-Villa, Marcel; Tempone, Antonio J; Telleria, Erich L; Ramalho-Ortigão, Marcelo; McMahon-Pratt, Diane; Traub-Csekö, Yara M

2015-03-01

Leishmaniasis is a serious problem that affects mostly poor countries. Various species of Leishmania are the agents of the disease, which take different clinical manifestations. The parasite is transmitted by sandflies, predominantly from the Phlebotomus genus in the Old World and Lutzomyia in the New World. During development in the gut, Leishmania must survive various challenges, which include avoiding being expelled with blood remnants after digestion. It is believed that attachment to the gut epithelium is a necessary step for vector infection, and molecules from parasites and sand flies have been implicated in this attachment. In previous work, monoclonal antibodies were produced against Leishmania. Among these an antibody was obtained against Leishmania braziliensis flagella, which blocked the attachment of Leishmania panamensis flagella to Phlebotomus papatasi guts. The protein recognized by this antibody was identified and named FLAG1, and the complete FLAG1 gene sequence was obtained. This protein was later independently identified as a small, myristoylated protein and called SMP1, so from now on it will be denominated FLAG1/SMP1. The FLAG1/SMP1 gene is expressed in all developmental stages of the parasite, but has higher expression in promastigotes. The anti-FLAG1/SMP1 antibody recognized the flagellum of all Leishmania species tested and generated the expected band by western blots. This antibody was used in attachment and infection blocking experiments. Using the New World vector Lutzomyia longipalpis and Leishmania infantum chagasi, no inhibition of attachment ex vivo or infection in vivo was seen. On the other hand, when the Old World vectors P. papatasi and Leishmania major were used, a significant decrease of both attachment and infection were seen in the presence of the antibody. We propose that FLAG1/SMP1 is involved in the attachment/infection of Leishmania in the strict vector P. papatasi and not the permissive vector L. longipalpis.

Combined Mass Spectrometry Imaging and Top-down Microproteomics Reveals Evidence of a Hidden Proteome in Ovarian Cancer.

PubMed

Delcourt, Vivian; Franck, Julien; Leblanc, Eric; Narducci, Fabrice; Robin, Yves-Marie; Gimeno, Jean-Pascal; Quanico, Jusal; Wisztorski, Maxence; Kobeissy, Firas; Jacques, Jean-François; Roucou, Xavier; Salzet, Michel; Fournier, Isabelle

2017-07-01

Recently, it was demonstrated that proteins can be translated from alternative open reading frames (altORFs), increasing the size of the actual proteome. Top-down mass spectrometry-based proteomics allows the identification of intact proteins containing post-translational modifications (PTMs) as well as truncated forms translated from reference ORFs or altORFs. Top-down tissue microproteomics was applied on benign, tumor and necrotic-fibrotic regions of serous ovarian cancer biopsies, identifying proteins exhibiting region-specific cellular localization and PTMs. The regions of interest (ROIs) were determined by MALDI mass spectrometry imaging and spatial segmentation. Analysis with a customized protein sequence database containing reference and alternative proteins (altprots) identified 15 altprots, including alternative G protein nucleolar 1 (AltGNL1) found in the tumor, and translated from an altORF nested within the GNL1 canonical coding sequence. Co-expression of GNL1 and altGNL1 was validated by transfection in HEK293 and HeLa cells with an expression plasmid containing a GNL1-FLAG (V5) construct. Western blot and immunofluorescence experiments confirmed constitutive co-expression of altGNL1-V5 with GNL1-FLAG. Taken together, our approach provides means to evaluate protein changes in the case of serous ovarian cancer, allowing the detection of potential markers that have never been considered. Copyright © 2017 The Author(s). Published by Elsevier B.V. All rights reserved.

Performance evaluation of the Sysmex XN-1000 hematology analyzer in assessment of the white blood cell count differential in pediatric specimens.

PubMed

Becker, P-H; Fenneteau, O; Da Costa, L

2016-02-01

The automated XN-1000 hematology analyzer enables to perform a blood cell count and a leukocyte differential. When abnormal cells were detected, a flag was generated by the analyzer and a manual microscopic examination of the corresponding blood film was performed. We compared the white blood cell differentials provided by the automated hematology analyzer XN-1000 in a pediatric population (n = 765) with those obtained through microscopic examination by cytologists and those obtained using a previous version of this analyzer, the XE-2100. Leukocytes count as well as flags sensitivity and specificity was analyzed. The leukocytes count provided by the analyzer is in good accordance with the differential obtained by manual count in children older than 3 months. The sensitivity for blast detection is 99% and the detection of reactive cells is 63%. The flag specificity remains low (<35%) for blood samples collected from infants between 8 days and 2 years of age, but increases up to 67% thereafter. The results obtained with the XN-1000 analyzer show an improvement in comparison with those obtained with the XE-2100 analyzer. The automated WBC differential provided by the XN-1000 analyzer in the pediatric setting is accurate, but a meticulous microscopic examination of blood smears remains necessary for infants up to 3 months of age to validate the analyzer flags. © 2015 John Wiley & Sons Ltd.

[Analysis of nuclear localization and signal function of MITF protein predisposing to Warrdenburg syndrome].

PubMed

Zhang, Hua; Feng, Juan; Chen, Hongsheng; Li, Jiada; Luo, Hunjin; Feng, Yong

2015-12-01

To study the role of dysfunction of nuclear localization signals (NLS) of MITF protein in the pathogenesis of Waardenburg syndrome. Eukaryotic expression plasmid pCMV-MITF-Flag was used as a template to generate mutant plasmid pCMV-MITF△NLS-Flag by molecular cloning technique in order to design the mutagenic primers. The UACC903 cells were transfected transiently with MITF and MITF△NLS plasmids, and the luciferase activity assays were performed to determine their impact on the transcriptional activities of target gene tyrosinase (TYR). The oligonucleotide 5'-GAACGAAGAAGAAGATTT-3' was subcloned into pEGFP-N1 to generate recombinant eukaryotic expression plasmid pEGFP-N1-MITF-NLS. The NIH3T3 cells were transfected separately with MITF, MITF△NLS, pEGFP-N1 and pEGFP-N1-NLS plasmids, and their subcellular distribution was observed by immunoflorescence assays. Expression plasmids for the mutant MITF△NLS with loss of core NLS sequence and pEGFP-N1-NLS coupled with MITF△NLS were successfully generated. Compared with the wild-type MITF, MITF△NLS was not able to transactivate the transcriptional activities of promoter TYR and did not affect the normal function of MITF. MITF△NLS was only localized in the cytoplasm and pEGFP-N1 was found in both the cytoplasm and nucleus, whereas pEGFP-N1-NLS was mainly located in the nucleus. This study has confirmed the localization function of NLS sequence 213ERRRRF218 within the MITF protein. Mutant MITF with loss of NLS has failed to transactivate the transcriptional activities of target gene TYR, which can result in melanocyte defects and cause WS.

The N-terminal region of the dopamine D2 receptor, a rhodopsin-like GPCR, regulates correct integration into the plasma membrane and endocytic routes

PubMed Central

Cho, DI; Min, C; Jung, KS; Cheong, SY; Zheng, M; Cheong, SJ; Oak, MH; Cheong, JH; Lee, BK; Kim, KM

2012-01-01

BACKGROUND AND PURPOSE Functional roles of the N-terminal region of rhodopsin-like GPCR family remain unclear. Using dopamine D2 and D3 receptors as a model system, we probed the roles of the N-terminal region in the signalling, intracellular trafficking of receptor proteins, and explored the critical factors that determine the functionality of the N-terminal region. EXPERIMENTAL APPROACH The N-terminal region of the D2 receptor was gradually shortened or switched with that of the D3 receptor or a non-specific sequence (FLAG), or potential N-terminal glycosylation sites were mutated. Effects of these manipulations on surface expression, internalization, post-endocytic behaviours and signalling were determined. KEY RESULTS Shortening the N-terminal region of the D2 receptor enhanced receptor internalization and impaired surface expression and signalling; ligand binding, desensitization and down-regulation were not affected but their association with a particular microdomain, caveolae, was disrupted. Replacement of critical residues within the N-terminal region with the FLAG epitope failed to restore surface expression but partially restored the altered internalization and signalling. When the N-terminal regions were switched between D2 and D3 receptors, cell surface expression pattern of each receptor was switched. Mutations of potential N-terminal glycosylation sites inhibited surface expression but enhanced internalization of D2 receptors. CONCLUSIONS AND IMPLICATIONS Shortening of N-terminus or mutation of glycosylation sites located within the N-terminus enhanced receptor internalization but impaired the surface expression of D2 receptors. The N-terminal region of the D2 receptor, in a sequence-specific manner, controls the receptor's conformation and integration into the plasma membrane, which determine its subcellular localization, intracellular trafficking and signalling properties. PMID:22117524

Endometase in Androgen-Repressed Human Prostate Cancer

DTIC Science & Technology

2005-03-01

immunosorbent assay (ELISA) and immunoblot using enhanced chemiluminescence (ECL). There are four specific endometase antibodies (Abs) available at the...lysates were An~t-FLAG-MJ2 analyzed by Western blotting using anti- FLAG-M2 antibody . Parental WT-MMP-26 EA-MMP-26 Parental (+YHJ-132) 100 125 54 51...invasion by MMP-26 antibodies is not due to the effects of the antibodies on cell attachment to extracellular matrix, cell proliferation, cytotoxicity

Isorhapontigenin (ISO) inhibited cell transformation by inducing G0/G1 phase arrest via increasing MKP-1 mRNA Stability.

PubMed

Gao, Guangxun; Chen, Liang; Li, Jingxia; Zhang, Dongyun; Fang, Yong; Huang, Haishan; Chen, Xiequn; Huang, Chuanshu

2014-05-15

The cancer chemopreventive property of Chinese herb new isolate isorhapontigenin (ISO) and mechanisms underlying its activity have never been explored. Here we demonstrated that ISO treatment with various concentrations for 3 weeks could dramatically inhibit TPA/EGF-induced cell transformation of Cl41 cells in Soft Agar assay, whereas co-incubation of cells with ISO at the same concentrations could elicit G0/G1 cell-cycle arrest without redundant cytotoxic effects on non-transformed cells. Further studies showed that ISO treatment resulted in cyclin D1 downregulation in dose- and time-dependent manner. Our results indicated that ISO regulated cyclin D1 at transcription level via targeting JNK/C-Jun/AP-1 activation. Moreover, we found that ISO-inhibited JNK/C-Jun/AP-1 activation was mediated by both upregulation of MKP-1 expression through increasing its mRNA stability and deactivating MKK7. Most importantly, MKP-1 knockdown could attenuate ISO-mediated suppression of JNK/C-Jun activation and cyclin D1 expression, as well as G0/G1 cell cycle arrest and cell transformation inhibition, while ectopic expression of FLAG-cyclin D1 T286A mutant also reversed ISO-induced G0/G1 cell-cycle arrest and inhibition of cell transformation. Our results demonstrated that ISO is a promising chemopreventive agent via upregulating mkp-1 mRNA stability, which is distinct from its cancer therapeutic effect with downregulation of XIAP and cyclin D1 expression.

Astaxanthin down-regulates Rad51 expression via inactivation of AKT kinase to enhance mitomycin C-induced cytotoxicity in human non-small cell lung cancer cells.

PubMed

Ko, Jen-Chung; Chen, Jyh-Cheng; Wang, Tai-Jing; Zheng, Hao-Yu; Chen, Wen-Ching; Chang, Po-Yuan; Lin, Yun-Wei

2016-04-01

Astaxanthin has been demonstrated to exhibit a wide range of beneficial effects, including anti-inflammatory and anti-cancer properties. However, the molecular mechanism of astaxanthin-induced cytotoxicity in non-small cell lung cancer (NSCLC) cells has not been identified. Rad51 plays a central role in homologous recombination, and studies show that chemo-resistant carcinomas exhibit high levels of Rad51 expression. In this study, astaxanthin treatment inhibited cell viability and proliferation of two NSCLC cells, A549 and H1703. Astaxanthin treatment (2.5-20 μM) decreased Rad51 expression and phospho-AKT(Ser473) protein level in a time and dose-dependent manner. Furthermore, expression of constitutively active AKT (AKT-CA) vector rescued the decreased Rad51 mRNA and protein levels in astaxanthin-treated NSCLC cells. Combined treatment with phosphatidylinositol 3-kinase (PI3K) inhibitors (LY294002 or wortmannin) further decreased the Rad51 expression in astaxanthin-exposed A549 and H1703 cells. Knockdown of Rad51 expression by transfection with si-Rad51 RNA or cotreatment with LY294002 further enhanced the cytotoxicity and cell growth inhibition of astaxanthin. Additionally, mitomycin C (MMC) as an anti-tumor antibiotic is widely used in clinical NSCLC chemotherapy. Combination of MMC and astaxanthin synergistically resulted in cytotoxicity and cell growth inhibition in NSCLC cells, accompanied with reduced phospho-AKT(Ser473) level and Rad51 expression. Overexpression of AKT-CA or Flag-tagged Rad51 reversed the astaxanthin and MMC-induced synergistic cytotoxicity. In contrast, pretreatment with LY294002 further decreased the cell viability in astaxanthin and MMC co-treated cells. In conclusion, astaxanthin enhances MMC-induced cytotoxicity by decreasing Rad51 expression and AKT activation. These findings may provide rationale to combine astaxanthin with MMC for the treatment of NSCLC. Copyright © 2016 Elsevier Inc. All rights reserved.

Identification of actin binding protein, ABP-280, as a binding partner of human Lnk adaptor protein.

PubMed

He, X; Li, Y; Schembri-King, J; Jakes, S; Hayashi, J

2000-08-01

Human Lnk (hLnk) is an adaptor protein with multiple functional domains that regulates T cell activation signaling. In order to identify cellular Lnk binding partners, a yeast two-hybrid screening of human spleen cDNA library was carried out using human hLnk as bait. A polypeptide sequence identical to the C-terminal segment of the actin binding protein (ABP-280) was identified as a hLnk binding protein. The expressed hLnk and the FLAG tagged C-terminal 673 amino acid residues of ABP-280 or the endogenous ABP-280 in COS-7 cells could be co-immunoprecipitated using antibodies either to hLnk, FLAG or ABP-280, respectively. Furthermore, immunofluorescence confocal microscope showed that hLnk and ABP-280 co-localized at the plasma membrane and at juxtanuclear region of COS-7 cells. In Jurkat cells, the endogenous hLnk also associates with the endogenous ABP-280 indicating that the association of these two proteins is physiological. The interacting domains of both proteins were mapped using yeast two-hybrid assays. Our results indicate that hLnk binds to the residues 2006-2454 (repeats 19-23C) of ABP-280. The domain in hLnk that associates with ABP-280 was mapped to an interdomain region of 56 amino acids between pleckstrin homology and Src homology 2 domains. These results suggest that hLnk may exert its regulatory role through its association with ABP-280.

Expression, purification and functional characterization of human equilibrative nucleoside transporter subtype-1 (hENT1) protein from Sf9 insect cells.

PubMed

Rehan, Shahid; Jaakola, Veli-Pekka

2015-10-01

Human equilibrative nucleoside transporter-1 (hENT1) is the major plasma membrane transporter involved in transportation of natural nucleosides as well as nucleoside analog drugs, used in anti-cancer and anti-viral therapies. Despite extensive biochemical and pharmacological studies, little is known about the structure-function relationship of this protein. The major obstacles to purification include a low endogenous expression level, the lack of an efficient expression and purification protocol, and the hydrophobic nature of the protein. Here, we report protein expression, purification and functional characterization of hENT1 from Sf9 insect cells. hENT1 expressed by Sf9 cells is functionally active as demonstrated by saturation binding with a Kd of 1.2±0.2nM and Bmax of 110±5pmol/mg for [(3)H]nitrobenzylmercaptopurine ribonucleoside ([(3)H]NBMPR). We also demonstrate purification of hENT1 using FLAG antibody affinity resin in lauryl maltose neopentyl glycol detergent with a Kd of 4.3±0.7nM. The yield of hENT1 from Sf9 cells was ∼0.5mg active transporter per liter of culture. The purified protein is functionally active, stable, homogenous and appropriate for further biophysical and structural studies. Copyright © 2015 Elsevier Inc. All rights reserved.

A Stable Human-Cell System Overexpressing Cystic Fibrosis Transmembrane Conductance Regulator Recombinant Protein at the Cell Surface

PubMed Central

Dai, Qun; Aleksandrov, Andrei A.; Bajrami, Bekim; Diego, Pamela Ann; Wu, Xing; Ray, Marjorie; Naren, Anjaparavanda P.; Riordan, John R.; Yao, Xudong; DeLucas, Lawrence J.; Urbatsch, Ina L.; Kappes, John C.

2015-01-01

Recent human clinical trials results demonstrated successful treatment for certain genetic forms of cystic fibrosis (CF). To extend treatment opportunities to those afflicted with other genetic forms of CF disease, structural and biophysical characterization of CF transmembrane conductance regulator (CFTR) is urgently needed. In this study, CFTR was modified with various tags, including a His10 purification tag, the SUMOstar (SUMO*) domain, an extracellular FLAG epitope, or an enhanced green fluorescent protein (EGFP), each alone or in various combinations. Expressed in HEK293 cells, recombinant CFTR proteins underwent complex glycosylation, compartmentalized with the plasma membrane, and exhibited regulated chloride-channel activity with only modest alterations in channel conductance and gating kinetics. Surface CFTR expression level was enhanced by the presence of SUMO* on the N-terminus. Quantitative mass-spectrometric analysis indicated approximately 10% of the total recombinant CFTR (SUMO*-CFTRFLAG-EGFP) localized to the plasma membrane. Trial purification using dodecylmaltoside for membrane protein extraction reproducibly recovered 178 ± 56 μg SUMO*-CFTRFLAG-EGFP per billion cells at 80% purity. Fluorescence size-exclusion chromatography indicated purified CFTR was monodisperse. These findings demonstrate a stable mammalian cell expression system capable of producing human CFTR of sufficient quality and quantity to augment futrure CF drug discovery efforts, including biophysical and structural studies. PMID:25577540

[Automated hematology analysers and spurious counts Part 3. Haemoglobin, red blood cells, cell count and indices, reticulocytes].

PubMed

Godon, Alban; Genevieve, Franck; Marteau-Tessier, Anne; Zandecki, Marc

2012-01-01

Several situations lead to abnormal haemoglobin measurement or to abnormal red blood cells (RBC) counts, including hyperlipemias, agglutinins and cryoglobulins, haemolysis, or elevated white blood cells (WBC) counts. Mean (red) cell volume may be also subject to spurious determination, because of agglutinins (mainly cold), high blood glucose level, natremia, anticoagulants in excess and at times technological considerations. Abnormality related to one measured parameter eventually leads to abnormal calculated RBC indices: mean cell haemoglobin content is certainly the most important RBC parameter to consider, maybe as important as flags generated by the haematology analysers (HA) themselves. In many circumstances, several of the measured parameters from cell blood counts (CBC) may be altered, and the discovery of a spurious change on one parameter frequently means that the validity of other parameters should be considered. Sensitive flags allow now the identification of several spurious counts, but only the most sophisticated HA have optimal flagging, and simpler ones, especially those without any WBC differential scattergram, do not share the same capacity to detect abnormal results. Reticulocytes are integrated into the CBC in many HA, and several situations may lead to abnormal counts, including abnormal gating, interference with intraerythrocytic particles, erythroblastosis or high WBC counts.

Alzheimer disease-related presenilin-1 variants exert distinct effects on monoamine oxidase-A activity in vitro.

PubMed

Pennington, Paul R; Wei, Zelan; Rui, Lewei; Doig, Jennifer A; Graham, Brett; Kuski, Kelly; Gabriel, Geraldine G; Mousseau, Darrell D

2011-07-01

Monoamine oxidase-A (MAO-A) has been associated with both depression and Alzheimer disease (AD). Recently, carriers of AD-related presenilin-1 (PS-1) alleles have been found to be at higher risk for developing clinical depression. We chose to examine whether PS-1 could influence MAO-A function in vitro. Overexpression of selected AD-related PS-1 variants (wildtype, Y115H, ΔEx9 and M146V) in mouse hippocampal HT-22 cells affects MAO-A catalytic activity in a variant-specific manner. The ability of the PS-1 substrate-competitor DAPT to induce MAO-A activity in cells expressing either PS-1 wildtype or PS-1(M146V) suggests the potential for a direct influence of PS-1 on MAO-A function. In support of this, we were able to co-immunoprecipitate MAO-A with FLAG-tagged PS-1 wildtype and M146V proteins. This potential for a direct protein-protein interaction between PS-1 and MAO-A is not specific for HT-22 cells as we were also able to co-immunoprecipitate MAO-A with FLAG-PS-1 variants in N2a mouse neuroblastoma cells and in HEK293 human embryonic kidney cells. Finally, we demonstrate that the two PS-1 variants reported to be associated with an increased incidence of clinical depression [e.g., A431E and L235V] both induce MAO-A activity in HT-22 cells. A direct influence of PS-1 variants on MAO-A function could provide an explanation for the changes in monoaminergic tone observed in several neurodegenerative processes including AD. The ability to induce MAO-A catalytic activity with a PS-1/γ-secretase inhibitor should also be considered when designing secretase inhibitor-based therapeutics.

Screening for Glycosylphosphatidylinositol-Modified Cell Wall Proteins in Pichia pastoris and Their Recombinant Expression on the Cell Surface

PubMed Central

Zhang, Li; Liang, Shuli; Zhou, Xinying; Jin, Zi; Jiang, Fengchun; Han, Shuangyan; Zheng, Suiping

2013-01-01

Glycosylphosphatidylinositol (GPI)-anchored glycoproteins have various intrinsic functions in yeasts and different uses in vitro. In the present study, the genome of Pichia pastoris GS115 was screened for potential GPI-modified cell wall proteins. Fifty putative GPI-anchored proteins were selected on the basis of (i) the presence of a C-terminal GPI attachment signal sequence, (ii) the presence of an N-terminal signal sequence for secretion, and (iii) the absence of transmembrane domains in mature protein. The predicted GPI-anchored proteins were fused to an alpha-factor secretion signal as a substitute for their own N-terminal signal peptides and tagged with the chimeric reporters FLAG tag and mature Candida antarctica lipase B (CALB). The expression of fusion proteins on the cell surface of P. pastoris GS115 was determined by whole-cell flow cytometry and immunoblotting analysis of the cell wall extracts obtained by β-1,3-glucanase digestion. CALB displayed on the cell surface of P. pastoris GS115 with the predicted GPI-anchored proteins was examined on the basis of potential hydrolysis of p-nitrophenyl butyrate. Finally, 13 proteins were confirmed to be GPI-modified cell wall proteins in P. pastoris GS115, which can be used to display heterologous proteins on the yeast cell surface. PMID:23835174

A Cleavable N-Terminal Signal Peptide Promotes Widespread Olfactory Receptor Surface Expression in HEK293T Cells

PubMed Central

Shepard, Blythe D.; Natarajan, Niranjana; Protzko, Ryan J.; Acres, Omar W.; Pluznick, Jennifer L.

2013-01-01

Olfactory receptors (ORs) are G protein-coupled receptors that detect odorants in the olfactory epithelium, and comprise the largest gene family in the genome. Identification of OR ligands typically requires OR surface expression in heterologous cells; however, ORs rarely traffic to the cell surface when exogenously expressed. Therefore, most ORs are orphan receptors with no known ligands. To date, studies have utilized non-cleavable rhodopsin (Rho) tags and/or chaperones (i.e. Receptor Transporting Protein, RTP1S, Ric8b and Gαolf) to improve surface expression. However, even with these tools, many ORs still fail to reach the cell surface. We used a test set of fifteen ORs to examine the effect of a cleavable leucine-rich signal peptide sequence (Lucy tag) on OR surface expression in HEK293T cells. We report here that the addition of the Lucy tag to the N-terminus increases the number of ORs reaching the cell surface to 7 of the 15 ORs (as compared to 3/15 without Rho or Lucy tags). Moreover, when ORs tagged with both Lucy and Rho were co-expressed with previously reported chaperones (RTP1S, Ric8b and Gαolf), we observed surface expression for all 15 receptors examined. In fact, two-thirds of Lucy-tagged ORs are able to reach the cell surface synergistically with chaperones even when the Rho tag is removed (10/15 ORs), allowing for the potential assessment of OR function with only an 8-amino acid Flag tag on the mature protein. As expected for a signal peptide, the Lucy tag was cleaved from the mature protein and did not alter OR-ligand binding and signaling. Our studies demonstrate that widespread surface expression of ORs can be achieved in HEK293T cells, providing promise for future large-scale deorphanization studies. PMID:23840901

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

Chen, Haiwen; Li, Hongliang, E-mail: honglianglity@sina.com; Chen, Qi

Docetaxel efficiency in the therapy of prostate cancer (PCa) patients is limited due to the development of chemoresistance. Recent studies have implied a role of INPP4B in tumor chemoresistance, while the effects of INPP4B on docetaxel resistance in PCa have not been elucidated. In the present study, the docetaxel-resistant human PCa cell lines PC3-DR and DU-145-DR were established from the parental cell lines PC3 and DU-145, and the expression and role of INPP4B in docetaxel-resistant PCa cells were investigated. The results demonstrated that INPP4B expression was significantly downregulated in docetaxel-resistant cells. Overexpression of INPP4B increased the sensitivity to docetaxel andmore » promoted cell apoptosis in PC3-DR and DU-145-DR cells. In addition, INPP4B overexpression downregulated the expression of the mesenchymal markers fibronectin, N-cadherin, and vimentin, and upregulated the expression level of the epithelial maker E-cadherin. Furthermore, INPP4B overexpression markedly inhibited the PI3K/Akt pathway. We also found that IGF-1, the inhibitor of PI3K/Akt, markedly blocked the change in EMT markers induced by overexpression of INPP4B, and reversed the resistance of PC3-DR and DU-145-DR cells to docetaxel, which is sensitized by Flag-INPP4B. In summary, the presented data indicate that INPP4B is crucial for docetaxel-resistant PCa cell survival, potentially by regulating EMT through the PI3K/Akt signaling pathway. - Highlights: • INPP4B is downregulated in docetaxel-resistant PCa cells. • INPP4B inhibits cell proliferation. • INPP4B induces cell apoptosis. • INPP4B inhibits PCa cell EMT.« less

The type 2 dengue virus envelope protein interacts with small ubiquitin-like modifier-1 (SUMO-1) conjugating enzyme 9 (Ubc9).

PubMed

Chiu, Mei-Wui; Shih, Hsiu-Ming; Yang, Tsung-Han; Yang, Yun-Liang

2007-05-01

Dengue viruses are mosquito-borne flaviviruses and may cause the life-threatening dengue hemorrhagic fever and dengue shock syndrome. Its envelope protein is responsible mainly for the virus attachment and entry to host cells. To identify the human cellular proteins interacting with the envelope protein of dengue virus serotype 2 inside host cells, we have performed a screening with the yeast-two-hybrid-based "Functional Yeast Array". Interestingly, the small ubiquitin-like modifier-1 conjugating enzyme 9 protein, modulating cellular processes such as those regulating signal transduction and cell growth, was one of the candidates interacting with the dengue virus envelope protein. With co-precipitation assay, we have demonstrated that it indeed could interact directly with the Ubc9 protein. Site-directed mutagenesis has demonstrated that Ubc9 might interact with the E protein via amino acid residues K51 and K241. Furthermore, immunofluorescence microscopy has shown that the DV2E-EGFP proteins tended to progress toward the nuclear membrane and co-localized with Flag-Ubc9 proteins around the nuclear membrane in the cytoplasmic side, and DV2E-EGFP also shifted the distribution of Flag-Ubc9 from evenly in the nucleus toward concentrating around the nuclear membrane in the nucleic side. In addition, over-expression of Ubc9 could reduce the plaque formation of the dengue virus in mammalian cells. This is the first report that DV envelope proteins can interact with the protein of sumoylation system and Ubc9 may involve in the host defense system to prevent virus propagation.

Interaction between p230 and MACF1 is associated with transport of a glycosyl phosphatidyl inositol-anchored protein from the Golgi to the cell periphery.

PubMed

Kakinuma, Takumi; Ichikawa, Haruo; Tsukada, Yoshito; Nakamura, Takashi; Toh, Ban-Hock

2004-08-15

The molecular basis by which proteins are transported along cytoskeletal tracts from the trans-Golgi network (TGN) to the cell periphery remains poorly understood. Previously, using human autoimmune sera, we identified and characterized a TGN protein, p230/Golgin-245, an extensively coiled-coil protein with flexible amino- and carboxyl-terminal ends, that is anchored to TGN membranes and TGN-derived vesicles by its carboxyl-terminal GRIP domain. To identify molecules that interact with the flexible amino-terminal end of p230, we used this domain as bait to screen a human brain cDNA library in a yeast two-hybrid assay. We found that this domain interacts with the carboxyl-terminal domain of MACF1, a protein that cross-links microtubules to the actin cytoskeleton. The interaction was confirmed by co-immunoprecipitation, an in vitro binding assay, double immunofluorescence images demonstrating overlapped localization in HeLa cells, and co-localization of FLAG-tagged constructs containing the interacting domains of these two proteins with their endogenous partners. Expression in HeLa cells of FLAG-tagged constructs containing the interacting domains of p230 and MACF1 disrupted transport of the glycosyl phosphatidyl inositol-anchored marker protein conjugated with yellow fluorescent protein (YFP-SP-GPI), while trafficking of the transmembrane marker protein, vesicular stomatitis virus glycoprotein conjugated with YFP (VSVG3-GL-YFP), was unaffected. Our results suggest that p230, through its interaction with MACF1, provides the molecular link for transport of GPI-anchored proteins along the microtubule and actin cytoskeleton from the TGN to the cell periphery.

Synthesis and materialization of a reaction-diffusion French flag pattern

NASA Astrophysics Data System (ADS)

Zadorin, Anton S.; Rondelez, Yannick; Gines, Guillaume; Dilhas, Vadim; Urtel, Georg; Zambrano, Adrian; Galas, Jean-Christophe; Estevez-Torres, André

2017-10-01

During embryo development, patterns of protein concentration appear in response to morphogen gradients. These patterns provide spatial and chemical information that directs the fate of the underlying cells. Here, we emulate this process within non-living matter and demonstrate the autonomous structuration of a synthetic material. First, we use DNA-based reaction networks to synthesize a French flag, an archetypal pattern composed of three chemically distinct zones with sharp borders whose synthetic analogue has remained elusive. A bistable network within a shallow concentration gradient creates an immobile, sharp and long-lasting concentration front through a reaction-diffusion mechanism. The combination of two bistable circuits generates a French flag pattern whose 'phenotype' can be reprogrammed by network mutation. Second, these concentration patterns control the macroscopic organization of DNA-decorated particles, inducing a French flag pattern of colloidal aggregation. This experimental framework could be used to test reaction-diffusion models and fabricate soft materials following an autonomous developmental programme.

A cCPE-based xenon biosensor for magnetic resonance imaging of claudin-expressing cells.

PubMed

Piontek, Anna; Witte, Christopher; May Rose, Honor; Eichner, Miriam; Protze, Jonas; Krause, Gerd; Piontek, Jörg; Schröder, Leif

2017-06-01

The majority of malignant tumors originate from epithelial cells, and many of them are characterized by an overexpression of claudins (Cldns) and their mislocalization out of tight junctions. We utilized the C-terminal claudin-binding domain of Clostridium perfringens enterotoxin (cCPE), with its high affinity to specific members of the claudin family, as the targeting unit for a claudin-sensitive cancer biosensor. To overcome the poor sensitivity of conventional relaxivity-based magnetic resonance imaging (MRI) contrast agents, we utilized the superior sensitivity of xenon Hyper-CEST biosensors. We labeled cCPE for both xenon MRI and fluorescence detection. As one readout module, we employed a cryptophane (CrA) monoacid and, as the second, a fluorescein molecule. Both were conjugated separately to a biotin molecule via a polyethyleneglycol chemical spacer and later via avidin linked to GST-cCPE. Nontransfected HEK293 cells and HEK293 cells stably expressing Cldn4-FLAG were incubated with the cCPE-based biosensor. Fluorescence-based flow cytometry and xenon MRI demonstrated binding of the biosensor specifically to Cldn4-expressing cells. This study provides proof of concept for the use of cCPE as a carrier for diagnostic contrast agents, a novel approach for potential detection of Cldn3/-4-overexpressing tumors for noninvasive early cancer detection. © 2017 New York Academy of Sciences.

Human α1β3γ2L gamma-aminobutyric acid type A receptors: High-level production and purification in a functional state.

PubMed

Dostalova, Zuzana; Zhou, Xiaojuan; Liu, Aiping; Zhang, Xi; Zhang, Yinghui; Desai, Rooma; Forman, Stuart A; Miller, Keith W

2014-02-01

Gamma-aminobutyric acid type A receptors (GABA(A)Rs) are the most important inhibitory chloride ion channels in the central nervous system and are major targets for a wide variety of drugs. The subunit compositions of GABA(A)Rs determine their function and pharmacological profile. GABAA Rs are heteropentamers of subunits, and (α1)2 (β3)2 (γ2L)1 is a common subtype. Biochemical and biophysical studies of GABA(A)Rs require larger quantities of receptors of defined subunit composition than are currently available. We previously reported high-level production of active human α1β3 GABA(A)R using tetracycline-inducible stable HEK293 cells. Here we extend the strategy to receptors containing three different subunits. We constructed a stable tetracycline-inducible HEK293-TetR cell line expressing human (N)-FLAG-α1β3γ2L-(C)-(GGS)3 GK-1D4 GABA(A)R. These cells achieved expression levels of 70-90 pmol [(3)H]muscimol binding sites/15-cm plate at a specific activity of 15-30 pmol/mg of membrane protein. Incorporation of the γ2 subunit was confirmed by the ratio of [(3)H]flunitrazepam to [(3)H]muscimol binding sites and sensitivity of GABA-induced currents to benzodiazepines and zinc. The α1β3γ2L GABA(A)Rs were solubilized in dodecyl-D-maltoside, purified by anti-FLAG affinity chromatography and reconstituted in CHAPS/asolectin at an overall yield of ∼ 30%. Typical purifications yielded 1.0-1.5 nmoles of [(3)H]muscimol binding sites/60 plates. Receptors with similar properties could be purified by 1D4 affinity chromatography with lower overall yield. The composition of the purified, reconstituted receptors was confirmed by ligand binding, Western blot, and proteomics. Allosteric interactions between etomidate and [(3)H]muscimol binding were maintained in the purified state. © 2013 The Protein Society.

[Expression of Dengue virus type 2 nonstructural protein 3 and isolation of host proteins interacting with it].

PubMed

Weng, Daihui; Lei, Yingfeng; Dong, Yangchao; Han, Peijun; Ye, Chuantao; Yang, Jing; Wang, Yuan; Yin, Wen

2015-12-01

To construct the plasmid expressing the fusion protein of Dengue virus type 2 (DENV2) nonstructural protein 3 (NS3) with affinity tag, and isolate the cellular proteins interacting with NS3 protein using tandem affinity purification (TAP) assay. Primers for amplifying NS3 gene were designed according to the sequence of DENV2 genome and chemically synthesized. The NS3 fragments, after amplified by PCR with DENV2 cDNA as template, were digested and cloned into the mammalian eukaryotic expression vector pCI-SF with the tandem affinity tag (FLAG-StrepII). The recombinant pCI-NS3-SF was transiently transformed by Lipofectamine(TM) 2000 into HEK293T cells, and the expression of the fusion protein was confirmed by Western blotting. Cellular proteins that interacted with NS3 were isolated and purified by TAP assay. The eukaryotic expression vector expressing NS3 protein was successfully constructed. The host proteins interacting with NS3 protein were isolated by TAP system. TAP is an efficient method to isolate the cellular proteins interacting with DENV2 NS3.

Characterization and expression patterns of key C4 photosynthetic pathway genes in bread wheat (Triticum aestivum L.) under field conditions.

PubMed

Bachir, Daoura Goudia; Saeed, Iqbal; Song, Quanhao; Linn, Tay Zar; Chen, Liang; Hu, Yin-Gang

2017-06-01

Wheat is a C 3 plant with relatively low photosynthetic efficiency and is a potential target for C 4 photosynthetic pathway engineering. Here we reported the characterization of four key C 4 pathway genes and assessed their expression patterns and enzymatic activities at three growth stages in flag leaves of 59 bread wheat genotypes. The C 4 -like genes homologous to PEPC, NADP-ME, MDH, and PPDK in maize were identified in the A, B, and D sub-genomes of bread wheat, located on the long arms of chromosomes 3 and 5 (TaPEPC), short arms of chromosomes 1 and 3 (TaNADP-ME), long arms of chromosomes 1 and 7 (TaMDH), and long arms of chromosome 1 (TaPPDK), respectively. All the four C 4 -like genes were expressed in the flag leaves at the three growth stages with considerable variations among the 59 bread wheat genotypes. Significant differences were observed between the photosynthesis rates (A) of wheat genotypes with higher expressions of TaPEPC_5, TaNADP-ME_1, and TaMDH_7 at heading and middle grain-filling stages and those with intermediate and low expressions. Our results also indicated that the four C 4 enzymes showed activity in the flag leaves and were obviously different among the 59 wheat genotypes. The activities of PEPcase and PPDK decreased at anthesis and slightly increased at grain-filling stage, while NADP-ME and MDH exhibited a decreasing trend at the three stages. The results of the current study could be very valuable and useful for wheat researchers in improving photosynthetic capacity of wheat. Copyright © 2017 Elsevier GmbH. All rights reserved.

The Myelin and Lymphocyte Protein MAL Is Required for Binding and Activity of Clostridium perfringens ε-Toxin

PubMed Central

Oo, Myat Lin; Anrather, Josef; Schaeren-Wiemers, Nicole; Alonso, Miguel A.; Fischetti, Vincent A.; McClain, Mark S.; Vartanian, Timothy

2015-01-01

Clostridium perfringens ε-toxin (ETX) is a potent pore-forming toxin responsible for a central nervous system (CNS) disease in ruminant animals with characteristics of blood-brain barrier (BBB) dysfunction and white matter injury. ETX has been proposed as a potential causative agent for Multiple Sclerosis (MS), a human disease that begins with BBB breakdown and injury to myelin forming cells of the CNS. The receptor for ETX is unknown. Here we show that both binding of ETX to mammalian cells and cytotoxicity requires the tetraspan proteolipid Myelin and Lymphocyte protein (MAL). While native Chinese Hamster Ovary (CHO) cells are resistant to ETX, exogenous expression of MAL in CHO cells confers both ETX binding and susceptibility to ETX-mediated cell death. Cells expressing rat MAL are ~100 times more sensitive to ETX than cells expressing similar levels of human MAL. Insertion of the FLAG sequence into the second extracellular loop of MAL abolishes ETX binding and cytotoxicity. ETX is known to bind specifically and with high affinity to intestinal epithelium, renal tubules, brain endothelial cells and myelin. We identify specific binding of ETX to these structures and additionally show binding to retinal microvasculature and the squamous epithelial cells of the sclera in wild-type mice. In contrast, there is a complete absence of ETX binding to tissues from MAL knockout (MAL-/-) mice. Furthermore, MAL-/- mice exhibit complete resistance to ETX at doses in excess of 1000 times the symptomatic dose for wild-type mice. We conclude that MAL is required for both ETX binding and cytotoxicity. PMID:25993478

The Myelin and Lymphocyte Protein MAL Is Required for Binding and Activity of Clostridium perfringens ε-Toxin.

PubMed

Rumah, Kareem Rashid; Ma, Yinghua; Linden, Jennifer R; Oo, Myat Lin; Anrather, Josef; Schaeren-Wiemers, Nicole; Alonso, Miguel A; Fischetti, Vincent A; McClain, Mark S; Vartanian, Timothy

2015-05-01

Clostridium perfringens ε-toxin (ETX) is a potent pore-forming toxin responsible for a central nervous system (CNS) disease in ruminant animals with characteristics of blood-brain barrier (BBB) dysfunction and white matter injury. ETX has been proposed as a potential causative agent for Multiple Sclerosis (MS), a human disease that begins with BBB breakdown and injury to myelin forming cells of the CNS. The receptor for ETX is unknown. Here we show that both binding of ETX to mammalian cells and cytotoxicity requires the tetraspan proteolipid Myelin and Lymphocyte protein (MAL). While native Chinese Hamster Ovary (CHO) cells are resistant to ETX, exogenous expression of MAL in CHO cells confers both ETX binding and susceptibility to ETX-mediated cell death. Cells expressing rat MAL are ~100 times more sensitive to ETX than cells expressing similar levels of human MAL. Insertion of the FLAG sequence into the second extracellular loop of MAL abolishes ETX binding and cytotoxicity. ETX is known to bind specifically and with high affinity to intestinal epithelium, renal tubules, brain endothelial cells and myelin. We identify specific binding of ETX to these structures and additionally show binding to retinal microvasculature and the squamous epithelial cells of the sclera in wild-type mice. In contrast, there is a complete absence of ETX binding to tissues from MAL knockout (MAL-/-) mice. Furthermore, MAL-/- mice exhibit complete resistance to ETX at doses in excess of 1000 times the symptomatic dose for wild-type mice. We conclude that MAL is required for both ETX binding and cytotoxicity.

21 CFR 864.5220 - Automated differential cell counter.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Automated differential cell counter. 864.5220... § 864.5220 Automated differential cell counter. (a) Identification. An automated differential cell... have the capability to flag, count, or classify immature or abnormal hematopoietic cells of the blood...

21 CFR 864.5220 - Automated differential cell counter.

Code of Federal Regulations, 2014 CFR

2014-04-01

... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Automated differential cell counter. 864.5220... § 864.5220 Automated differential cell counter. (a) Identification. An automated differential cell... have the capability to flag, count, or classify immature or abnormal hematopoietic cells of the blood...

21 CFR 864.5220 - Automated differential cell counter.

Code of Federal Regulations, 2012 CFR

2012-04-01

... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Automated differential cell counter. 864.5220... § 864.5220 Automated differential cell counter. (a) Identification. An automated differential cell... have the capability to flag, count, or classify immature or abnormal hematopoietic cells of the blood...

21 CFR 864.5220 - Automated differential cell counter.

Code of Federal Regulations, 2013 CFR

2013-04-01

... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Automated differential cell counter. 864.5220... § 864.5220 Automated differential cell counter. (a) Identification. An automated differential cell... have the capability to flag, count, or classify immature or abnormal hematopoietic cells of the blood...

21 CFR 864.5220 - Automated differential cell counter.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Automated differential cell counter. 864.5220... § 864.5220 Automated differential cell counter. (a) Identification. An automated differential cell... have the capability to flag, count, or classify immature or abnormal hematopoietic cells of the blood...

Mechanism of Tumor Metastasis Suppression by the KAI1 Gene

DTIC Science & Technology

2008-02-01

antibody to Flag covalently crosslinked to agarose beads followed by western blot with monoclonal antibody to Flag (lanes 1, 2). For coimmunoprecipitation...negative controls (lanes 4, 6). IgH appeared in lanes 5 and 6, as antibody to hemagglutinin was not crosslinked to the agarose beads during...mixed in the presence of a cell-impermeable crosslinker DTSSP for 30 min followed by immunoprecipitation with DARC antibody and western blot with

RITA inhibits growth of human hepatocellular carcinoma through induction of apoptosis.

PubMed

Wang, Haihe; Chen, Guofu; Wang, Hongzhi; Liu, Chunbo

2013-01-01

RBP-J-interacting and tubulin-associated (RITA) is a novel RBP-J-interacting protein that downregulates Notch-mediated transcription. The current study focuses on the antitumor effect of RITA in human hepatocellular carcinoma (HCC) and aims to explore its molecular mechanism. Thirty paired HCC and adjacent non-tumoral liver samples were analyzed by real-time quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). RITA overexpression was induced by transfection of a pcDNA3.1-Flag-RITA plasmid into HepG2 cells. RITA knockdown was achieved by siRNA transfection. mRNA and protein expression of target genes were quantified by qRT-PCR and Western blotting, respectively. Cell proliferation and apoptosis were measured using MTT assay and flow cytometry. Our results demonstrate that adjacent nontumoral liver samples exhibited increased RITA expression compared to HCC tissues (p < 0.05); RITA levels were associated with tumor differentiation status. Overexpression of RITA suppressed cell proliferation and promoted early apoptosis, while its silencing promoted cell growth dramatically (p < 0.05). RITA overexpression upregulated p53 and reduced cyclin E levels, whereas silencing of RITA had the opposite effect on p53 and cyclin E expression. Our in vitro results represent the first evidence that RITA might suppress tumor growth and induce apoptosis in HCCs, and may be a potent antitumoral agent for HCC treatment that deserves further exploration.

Iridovirus CARD Protein Inhibits Apoptosis through Intrinsic and Extrinsic Pathways

PubMed Central

Chen, Chien-Wen; Wu, Ming-Shan; Huang, Yi-Jen; Lin, Pei-Wen; Shih, Chueh-Ju; Lin, Fu-Pang; Chang, Chi-Yao

2015-01-01

Grouper iridovirus (GIV) belongs to the genus Ranavirus of the family Iridoviridae; the genomes of such viruses contain an anti-apoptotic caspase recruitment domain (CARD) gene. The GIV-CARD gene encodes a protein of 91 amino acids with a molecular mass of 10,505 Daltons, and shows high similarity to other viral CARD genes and human ICEBERG. In this study, we used Northern blot to demonstrate that GIV-CARD transcription begins at 4 h post-infection; furthermore, we report that its transcription is completely inhibited by cycloheximide but not by aphidicolin, indicating that GIV-CARD is an early gene. GIV-CARD-EGFP and GIV-CARD-FLAG recombinant proteins were observed to translocate from the cytoplasm into the nucleus, but no obvious nuclear localization sequence was observed within GIV-CARD. RNA interference-mediated knockdown of GIV-CARD in GK cells infected with GIV inhibited expression of GIV-CARD and five other viral genes during the early stages of infection, and also reduced GIV infection ability. Immunostaining was performed to show that apoptosis was effectively inhibited in cells expressing GIV-CARD. HeLa cells irradiated with UV or treated with anti-Fas antibody will undergo apoptosis through the intrinsic and extrinsic pathways, respectively. However, over-expression of recombinant GIV-CARD protein in HeLa cells inhibited apoptosis induced by mitochondrial and death receptor signaling. Finally, we report that expression of GIV-CARD in HeLa cells significantly reduced the activities of caspase-8 and -9 following apoptosis triggered by anti-Fas antibody. Taken together, these results demonstrate that GIV-CARD inhibits apoptosis through both intrinsic and extrinsic pathways. PMID:26047333

Department of Defense: Electronic Biometric Transmission Specification. Version 2.0

DTIC Science & Technology

2009-03-27

Abstractions = ABSTRACT Insignias & Symbols = SYMBOL Other Images = OTHER Information Item Number: 3 Tattoo Subclass Description: This information item...Tattoo Subclasses: American Flag = USA State Flag = STATE Nazi Flag = NAZI Confederate Flag = CONFED British Flag = BRIT Miscellaneous Flags = MFLAG...Vegetables = MPLANT Flag Tattoo Subclasses: American Flag = USA State Flag = STATE Nazi Flag = NAZI Confederate Flag = CONFED British Flag = BRIT

Tandem Affinity Purification of Protein Complexes from Eukaryotic Cells.

PubMed

Ma, Zheng; Fung, Victor; D'Orso, Iván

2017-01-26

The purification of active protein-protein and protein-nucleic acid complexes is crucial for the characterization of enzymatic activities and de novo identification of novel subunits and post-translational modifications. Bacterial systems allow for the expression and purification of a wide variety of single polypeptides and protein complexes. However, this system does not enable the purification of protein subunits that contain post-translational modifications (e.g., phosphorylation and acetylation), and the identification of novel regulatory subunits that are only present/expressed in the eukaryotic system. Here, we provide a detailed description of a novel, robust, and efficient tandem affinity purification (TAP) method using STREP- and FLAG-tagged proteins that facilitates the purification of protein complexes with transiently or stably expressed epitope-tagged proteins from eukaryotic cells. This protocol can be applied to characterize protein complex functionality, to discover post-translational modifications on complex subunits, and to identify novel regulatory complex components by mass spectrometry. Notably, this TAP method can be applied to study protein complexes formed by eukaryotic or pathogenic (viral and bacterial) components, thus yielding a wide array of downstream experimental opportunities. We propose that researchers working with protein complexes could utilize this approach in many different ways.

Transcriptome and secretome analyses of Phanerochaete chrysosporium reveal complex patterns of gene expression

Treesearch

Amber J. Vanden Wymelenberg; Jill A. Gaskell; Michael D. Mozuch; Philip J. Kersten; Grzegorz Sabat; Diego Martinez; Daniel Cullen

2009-01-01

The wood decay basidiomycete Phanerochaete chrysosporium was grown under standard ligninolytic or cellulolytic conditions and subjected to whole-genome expression microarray analysis and liquid chromatography-tandem mass spectrometry of extracellular proteins. A total of 545 genes were flagged on the basis of significant changes in transcript accumulation and/or...

Algorithms on Flag Manifolds for Knowledge Discovery in N-way Arrays

DTIC Science & Technology

2015-11-20

that three of 18 subjects will become symptomatic after only 8 hours. Host pathway analysis of a human endotoxin gene expression data set revealed a 14...pathway analysis of a human endotoxin gene expression data set revealed a 14 pathway signature that identified symptomatic subjects within 2-3 hours post

Construction of a functional silk-based biomaterial complex with immortalized chondrocytes in vivo.

PubMed

Ni, Yusu; Jiang, Yi; Wen, Jianchuan; Shao, Zhenzhong; Chen, Xin; Sun, Shan; Yu, Huiqian; Li, Wen

2014-04-01

To explore the feasibility of constructing a functional biomaterial complex with regenerated silk fibroin membrane and immortalized chondrocytes in vivo. Rat auricular chondrocytes (RACs) were transfected with the lentivirus vector pGC-FU-hTERT-3FLAG or pGC-FU-GFP-3FLAG, encoding the human telomerase reverse transcriptase (hTERT) or GFP gene. The effects of regenerated silk fibroin film on the adhesion, growth of immortalized chondrocytes and expression of collagen II in vitro were analyzed with immunofluorescent histochemistry. Immortalized RACs were transformed. Induction by nutrient medium promoted higher expression levels of collagen II in transformed chondrocytes. The regenerated silk fibroin film was not cytotoxic to immortalized chondrocytes and had no adverse influence on their adhesion. Collagen II expression was good in the immortalized chondrocytes in vivo. The construction of a silk-based biomaterial complex with immortalized chondrocytes may provide a feasible kind of functional biomaterial for the repair of cartilage defects in clinical applications. Copyright © 2013 Wiley Periodicals, Inc.

Targeted Delivery of TrkB Receptor to Phrenic Motoneurons Enhances Functional Recovery of Rhythmic Phrenic Activity after Cervical Spinal Hemisection

PubMed Central

Gransee, Heather M.; Zhan, Wen-Zhi; Sieck, Gary C.; Mantilla, Carlos B.

2013-01-01

Progressive recovery of rhythmic phrenic activity occurs over time after a spinal cord hemisection involving unilateral transection of anterolateral funiculi at C2 (SH). Brain-derived neurotrophic factor (BDNF) acting through its full-length tropomyosin related kinase receptor subtype B (TrkB.FL) contributes to neuroplasticity after spinal cord injury, but the specific cellular substrates remain unclear. We hypothesized that selectively targeting increased TrkB.FL expression to phrenic motoneurons would be sufficient to enhance recovery of rhythmic phrenic activity after SH. Several adeno-associated virus (AAV) serotypes expressing GFP were screened to determine specificity for phrenic motoneuron transduction via intrapleural injection in adult rats. GFP expression was present in the cervical spinal cord 3 weeks after treatment with AAV serotypes 7, 8, and 9, but not with AAV2, 6, or rhesus-10. Overall, AAV7 produced the most consistent GFP expression in phrenic motoneurons. SH was performed 3 weeks after intrapleural injection of AAV7 expressing human TrkB.FL-FLAG or saline. Delivery of TrkB.FL-FLAG to phrenic motoneurons was confirmed by FLAG protein expression in the phrenic motor nucleus and human TrkB.FL mRNA expression in microdissected phrenic motoneurons. In all SH rats, absence of ipsilateral diaphragm EMG activity was confirmed at 3 days post-SH, verifying complete interruption of ipsilateral descending drive to phrenic motoneurons. At 14 days post-SH, all AAV7-TrkB.FL treated rats (n = 11) displayed recovery of ipsilateral diaphragm EMG activity compared to 3 out of 8 untreated SH rats (p<0.01). During eupnea, AAV7-TrkB.FL treated rats exhibited 73±7% of pre-SH root mean squared EMG vs. only 31±11% in untreated SH rats displaying recovery (p<0.01). This study provides direct evidence that increased TrkB.FL expression in phrenic motoneurons is sufficient to enhance recovery of ipsilateral rhythmic phrenic activity after SH, indicating that selectively targeting gene expression in spared motoneurons below the level of spinal cord injury may promote functional recovery. PMID:23724091

The novel putative bile acid transporter SLC10A5 is highly expressed in liver and kidney

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

Fernandes, Carla F.; Godoy, Jose R.; Doering, Barbara

2007-09-14

Here we report the identification, cloning, and characterization of SLC10A5, which is a new member of Solute Carrier Family 10 (SLC10), also known as the 'sodium/bile acid cotransporter family'. Expression of SLC10A5/Slc10a5 was examined by quantitative real-time PCR and revealed its highest expression levels in liver and kidney in humans, rat and mouse. In rat liver and kidney, Slc10a5 expression was localized by in situ hybridization to hepatocytes and proximal tubules, respectively. A SLC10A5-FLAG fusion protein was expressed in HEK293 cells and showed an apparent molecular weight of 42 kDa after immunoprecipitation. When expressed in Xenopus laevis oocytes, the SLC10A5-FLAGmore » protein was detected in the oocyte's plasma membrane but showed no transport activity for taurocholate, cholate, estrone-3-sulfate, or dehydroepiandrosterone sulfate. As bile acid carriers are the most related carriers to SLC10A5 though, we strongly suppose that SLC10A5 is an orphan carrier with yet non-identified substrates.« less

Orientia tsutsugamushi Strain Ikeda Ankyrin Repeat-Containing Proteins Recruit SCF1 Ubiquitin Ligase Machinery via Poxvirus-Like F-Box Motifs.

PubMed

Beyer, Andrea R; VieBrock, Lauren; Rodino, Kyle G; Miller, Daniel P; Tegels, Brittney K; Marconi, Richard T; Carlyon, Jason A

2015-10-01

A rising theme among intracellular microbes is the delivery of ankyrin repeat-containing effectors (Anks) that interact with target proteins to co-opt host cell functions. Orientia tsutsugamushi, an obligate intracellular bacterium and the etiologic agent of scrub typhus, encodes one of the largest Ank repertoires of any sequenced microorganism. They have been previously identified as type 1 secretion system substrates. Here, in silico and manual sequence analyses revealed that a large proportion of O. tsutsugamushi strain Ikeda Anks bear a eukaryotic/poxvirus-like F-box motif, which is known to recruit host cell SCF1 ubiquitin ligase machinery. We assessed the Anks for the ability to serve as F-box proteins. Coimmunoprecipitation assays demonstrated that F-box-containing Anks interact with overexpressed and/or endogenous SCF1 components. When coexpressed with FLAG-Ank4_01 or FLAG-Ank9, a glutathione S-transferase (GST)-tagged version of the SCF1 component SKP1 localized to subcellular sites of FLAG-Ank accumulation. The abilities of recombinant Anks to interact and colocalize with SKP1 were F-box dependent. GST-SKP1 precipitated O. tsutsugamushi-derived Ank9 from infected host cells, verifying both that the pathogen expresses Ank9 during infection and the protein's capability to bind SKP1. Aligning O. tsutsugamushi, poxviral, and eukaryotic F-box sequences delineated three F-box residues that are highly conserved and likely to be functionally important. Substitution of these residues ablated the ability of GFP-Ank9 to interact with GST-SKP1. These results demonstrate that O. tsutsugamushi strain Ikeda Anks can co-opt host cell polyubiquitination machinery, provide the first evidence that an O. tsutsugamushi Ank does so during infection, and advance overall understanding of microbial F-box proteins. Ankyrin repeat-containing proteins (Anks) are important virulence factors of intracellular bacteria that mediate protein-protein interactions with host cell targets. Orientia tsutsugamushi, which causes a debilitating infection called scrub typhus in one of the most densely populated regions of the world, encodes one of the largest Ank armamentariums of any sequenced bacterium. This study demonstrates that O. tsutsugamushi strain Ikeda Anks also bear F-box motifs that interact with host cell polyubiquitination machinery. By proving that an Orientia-derived Ank interacts with SKP1 in infected cells, this evidences the first bona fide Orientia effector and the first example of an endogenous F-box-containing Ank-mammalian-host ligand interaction for any intracellular bacterium. Also, importantly, this work identifies key residues that are essential for microbial F-box function. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Orientia tsutsugamushi Strain Ikeda Ankyrin Repeat-Containing Proteins Recruit SCF1 Ubiquitin Ligase Machinery via Poxvirus-Like F-Box Motifs

PubMed Central

Beyer, Andrea R.; VieBrock, Lauren; Rodino, Kyle G.; Miller, Daniel P.; Tegels, Brittney K.; Marconi, Richard T.

2015-01-01

ABSTRACT A rising theme among intracellular microbes is the delivery of ankyrin repeat-containing effectors (Anks) that interact with target proteins to co-opt host cell functions. Orientia tsutsugamushi, an obligate intracellular bacterium and the etiologic agent of scrub typhus, encodes one of the largest Ank repertoires of any sequenced microorganism. They have been previously identified as type 1 secretion system substrates. Here, in silico and manual sequence analyses revealed that a large proportion of O. tsutsugamushi strain Ikeda Anks bear a eukaryotic/poxvirus-like F-box motif, which is known to recruit host cell SCF1 ubiquitin ligase machinery. We assessed the Anks for the ability to serve as F-box proteins. Coimmunoprecipitation assays demonstrated that F-box-containing Anks interact with overexpressed and/or endogenous SCF1 components. When coexpressed with FLAG-Ank4_01 or FLAG-Ank9, a glutathione S-transferase (GST)-tagged version of the SCF1 component SKP1 localized to subcellular sites of FLAG-Ank accumulation. The abilities of recombinant Anks to interact and colocalize with SKP1 were F-box dependent. GST-SKP1 precipitated O. tsutsugamushi-derived Ank9 from infected host cells, verifying both that the pathogen expresses Ank9 during infection and the protein's capability to bind SKP1. Aligning O. tsutsugamushi, poxviral, and eukaryotic F-box sequences delineated three F-box residues that are highly conserved and likely to be functionally important. Substitution of these residues ablated the ability of GFP-Ank9 to interact with GST-SKP1. These results demonstrate that O. tsutsugamushi strain Ikeda Anks can co-opt host cell polyubiquitination machinery, provide the first evidence that an O. tsutsugamushi Ank does so during infection, and advance overall understanding of microbial F-box proteins. IMPORTANCE Ankyrin repeat-containing proteins (Anks) are important virulence factors of intracellular bacteria that mediate protein-protein interactions with host cell targets. Orientia tsutsugamushi, which causes a debilitating infection called scrub typhus in one of the most densely populated regions of the world, encodes one of the largest Ank armamentariums of any sequenced bacterium. This study demonstrates that O. tsutsugamushi strain Ikeda Anks also bear F-box motifs that interact with host cell polyubiquitination machinery. By proving that an Orientia-derived Ank interacts with SKP1 in infected cells, this evidences the first bona fide Orientia effector and the first example of an endogenous F-box-containing Ank–mammalian-host ligand interaction for any intracellular bacterium. Also, importantly, this work identifies key residues that are essential for microbial F-box function. PMID:26170417

Establishment of a Conditional Transgenic Mouse Model Recapitulating EML4-ALK-Positive Human Non-Small Cell Lung Cancer.

PubMed

Pyo, Kyoung Ho; Lim, Sun Min; Kim, Hye Ryun; Sung, Young Hoon; Yun, Mi Ran; Kim, Sung-Moo; Kim, Hwan; Kang, Han Na; Lee, Ji Min; Kim, Sang Gyun; Park, Chae Won; Chang, Hyun; Shim, Hyo Sup; Lee, Han-Woong; Cho, Byoung Chul

2017-03-01

Anaplastic lymphoma receptor tyrosine kinase gene (ALK) fusion is a distinct molecular subclassification of NSCLC that is targeted by anaplastic lymphoma kinase (ALK) inhibitors. We established a transgenic mouse model that expresses tumors highly resembling human NSCLC harboring echinoderm microtubule associated protein like 4 gene (EML)-ALK fusion. We aimed to test an EML4-ALK transgenic mouse model as a platform for assessing the efficacy of ALK inhibitors and examining mechanisms of acquired resistance to ALK inhibitors. Transgenic mouse lines harboring LoxP-STOP-LoxP-FLAGS-tagged human EML4-ALK (variant 1) transgene was established by using C57BL/6N mice. The transgenic mouse model with highly lung-specific, inducible expression of echinoderm microtubule associated protein like 4-ALK fusion protein was established by crossing the EML4-ALK transgenic mice with mice expressing Cre-estrogen receptor fusion protein under the control of surfactant protein C gene (SPC). Expression of EML4-ALK transgene was induced by intraperitoneally injecting mice with tamoxifen. When the lung tumor of the mice treated with the ALK inhibitor crizotinib for 2 weeks was measured, tumor shrinkage was observed. EML4-ALK tumor developed after 1 week of tamoxifen treatment. Echinoderm microtubule associated protein like 4-ALK was strongly expressed in the lung but not in other organs. ALK and FLAGS expressions were observed by immunohistochemistry. Treatment of EML4-ALK tumor-bearing mice with crizotinib for 2 weeks induced dramatic shrinkage of tumors with no signs of toxicity. Furthermore, prolonged treatment with crizotinib led to acquired resistance in tumors, resulting in regrowth and disease progression. The resistant tumor nodules revealed acquired ALK G1202R mutations. An EML4-ALK transgenic mouse model for study of drug resistance was successfully established with short duration of tumorigenesis. This model should be a strong preclinical model for testing efficacy of ALK TKIs, providing a useful tool for investigating the mechanisms of acquired resistance and pursuing novel treatment strategies in ALK-positive lung cancer. Copyright © 2016 International Association for the Study of Lung Cancer. Published by Elsevier Inc. All rights reserved.

HNRNPLL stabilizes mRNAs for DNA replication proteins and promotes cell cycle progression in colorectal cancer cells.

PubMed

Sakuma, Keiichiro; Sasaki, Eiichi; Kimura, Kenya; Komori, Koji; Shimizu, Yasuhiro; Yatabe, Yasushi; Aoki, Masahiro

2018-06-05

HNRNPLL (heterogeneous nuclear ribonucleoprotein L-like), an RNA-binding protein that regulates alternative splicing of pre-mRNAs, has been shown to regulate differentiation of lymphocytes, as well as metastasis of colorectal cancer cells. Here we show that HNRNPLL promotes cell cycle progression and hence proliferation of colorectal cancer cells. Functional annotation analysis of those genes whose expression levels were changed by three-fold or more in RNA sequencing analysis between SW480 cells overexpressing HNRNPLL and those knocked down for HNRNPLL revealed enrichment of DNA replication-related genes by HNRNPLL overexpression. Among 13 genes detected in the DNA replication pathway, PCNA, RFC3, and FEN1 showed reproducible upregulation by HNRNPLL overexpression both at mRNA and protein levels in SW480 and HT29 cells. Importantly, knockdown of any of these genes alone suppressed the proliferation promoting effect induced by HNRNPLL overexpression. RNA-immunoprecipitation assay presented a binding of FLAG-tagged HNRNPLL to mRNA of these genes, and HNRNPLL overexpression significantly suppressed the downregulation of these genes during 12 hours of actinomycin D treatment, suggesting a role of HNRNPLL in mRNA stability. Finally, analysis of a public RNA sequencing dataset of clinical samples suggested a link between overexpression of HNRNPLL and that of PCNA, RFC3, and FEN1. This link was further supported by immunohistochemistry of colorectal cancer clinical samples, whereas expression of CDKN1A, which is known to inhibit the cooperative function of PCNA, RFC3, and FEN1, was negatively associated with HNRNPLL expression. These results indicate that HNRNPLL stabilizes mRNAs encoding regulators of DNA replication and promotes colorectal cancer cell proliferation. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Molecular determinants of the olfactory receptor Olfr544 activation by azelaic acid.

PubMed

Thach, Trung Thanh; Hong, Yu-Jung; Lee, Sangho; Lee, Sung-Joon

2017-04-01

The mouse olfactory receptor Olfr544 is expressed in several non-olfactory tissues and has been suggested as a functional receptor regulating different signaling pathways. However, the molecular interaction between Olfr544 and its natural ligand, azelaic acid (AzA), remains poorly characterized, primarily due to difficulties in the heterologous expression of the receptor protein on the cell membrane and lack of entire protein structure. In this report, we describe the molecular determinants of Olfr544 activation by AzA. N-terminal lucy-flag-rho tag ensured the heterologous expression of Olfr544 on the Hana3A cell surface. Molecular modeling and docking combined with mutational analysis identified amino acid residues in the Olfr544 for the interaction with AzA. Our data demonstrated that the Y109 residue in transmembrane helix 3 forms a hydrogen bond with AzA, which is crucial for the receptor-ligand interaction and activation. Y109 is required for the Olfr544 activation by AzA which, in turn, stimulates the Olfr544-dependent CREB-PGC-1α signaling axis and is followed by the induction of mitochondrial biogenesis in Olfr544 wild-type transfected Hana3A cells, but not in mock or Y109A mutant transfected cells. Collectively, these data indicated that a hydrogen bond between Y109 residue and AzA is a major determinant of the Olfr544-AzA interaction and activation. Copyright © 2017 Elsevier Inc. All rights reserved.

Immunofluorescence Analysis of Endogenous and Exogenous Centromere-kinetochore Proteins

PubMed Central

Niikura, Yohei; Kitagawa, Katsumi

2016-01-01

"Centromeres" and "kinetochores" refer to the site where chromosomes associate with the spindle during cell division. Direct visualization of centromere-kinetochore proteins during the cell cycle remains a fundamental tool in investigating the mechanism(s) of these proteins. Advanced imaging methods in fluorescence microscopy provide remarkable resolution of centromere-kinetochore components and allow direct observation of specific molecular components of the centromeres and kinetochores. In addition, methods of indirect immunofluorescent (IIF) staining using specific antibodies are crucial to these observations. However, despite numerous reports about IIF protocols, few discussed in detail problems of specific centromere-kinetochore proteins.1-4 Here we report optimized protocols to stain endogenous centromere-kinetochore proteins in human cells by using paraformaldehyde fixation and IIF staining. Furthermore, we report protocols to detect Flag-tagged exogenous CENP-A proteins in human cells subjected to acetone or methanol fixation. These methods are useful in detecting and quantifying endogenous centromere-kinetochore proteins and Flag-tagged CENP-A proteins, including those in human cells. PMID:26967065

Genome-wide analysis of murine renal distal convoluted tubular cells for the target genes of mineralocorticoid receptor

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

Ueda, Kohei; Fujiki, Katsunori; Shirahige, Katsuhiko

Highlights: • We define a target gene of MR as that with MR-binding to the adjacent region of DNA. • We use ChIP-seq analysis in combination with microarray. • We, for the first time, explore the genome-wide binding profile of MR. • We reveal 5 genes as the direct target genes of MR in the renal epithelial cell-line. - Abstract: Background and objective: Mineralocorticoid receptor (MR) is a member of nuclear receptor family proteins and contributes to fluid homeostasis in the kidney. Although aldosterone-MR pathway induces several gene expressions in the kidney, it is often unclear whether the gene expressionsmore » are accompanied by direct regulations of MR through its binding to the regulatory region of each gene. The purpose of this study is to identify the direct target genes of MR in a murine distal convoluted tubular epithelial cell-line (mDCT). Methods: We analyzed the DNA samples of mDCT cells overexpressing 3xFLAG-hMR after treatment with 10{sup −7} M aldosterone for 1 h by chromatin immunoprecipitation with deep-sequence (ChIP-seq) and mRNA of the cell-line with treatment of 10{sup −7} M aldosterone for 3 h by microarray. Results: 3xFLAG-hMR overexpressed in mDCT cells accumulated in the nucleus in response to 10{sup −9} M aldosterone. Twenty-five genes were indicated as the candidate target genes of MR by ChIP-seq and microarray analyses. Five genes, Sgk1, Fkbp5, Rasl12, Tns1 and Tsc22d3 (Gilz), were validated as the direct target genes of MR by quantitative RT-qPCR and ChIP-qPCR. MR binding regions adjacent to Ctgf and Serpine1 were also validated. Conclusions: We, for the first time, captured the genome-wide distribution of MR in mDCT cells and, furthermore, identified five MR target genes in the cell-line. These results will contribute to further studies on the mechanisms of kidney diseases.« less

The Full Kostant-Toda Hierarchy on the Positive Flag Variety

NASA Astrophysics Data System (ADS)

Kodama, Yuji; Williams, Lauren

2015-04-01

We study some geometric and combinatorial aspects of the solution to the full Kostant-Toda (f-KT) hierarchy, when the initial data is given by an arbitrary point on the totally non-negative (tnn) flag variety of . The f-KT flows on the tnn flag variety are complete, and we show that their asymptotics are completely determined by the cell decomposition of the tnn flag variety given by Rietsch (Total positivity and real flag varieties. Ph.D. Thesis, Massachusetts Institute of Technology, Cambridge, 1998). Our results represent the first results on the asymptotics of the f-KT hierarchy (and even the f-KT lattice); moreover, our results are not confined to the generic flow, but cover non-generic flows as well. We define the f-KT flow on the weight space via the moment map, and show that the closure of each f-KT flow forms an interesting convex polytope which we call a Bruhat interval polytope. In particular, the Bruhat interval polytope for the generic flow is the permutohedron of the symmetric group . We also prove analogous results for the full symmetric Toda hierarchy, by mapping our f-KT solutions to those of the full symmetric Toda hierarchy. In the appendix we show that Bruhat interval polytopes are generalized permutohedra, in the sense of Postnikov (Int. Math. Res. Not. IMRN (6):1026-1106, 2009).

Identification of Human P2X1 Receptor-interacting Proteins Reveals a Role of the Cytoskeleton in Receptor Regulation*

PubMed Central

Lalo, Ulyana; Roberts, Jonathan A.; Evans, Richard J.

2011-01-01

P2X1 receptors are ATP-gated ion channels expressed by smooth muscle and blood cells. Carboxyl-terminally His-FLAG-tagged human P2X1 receptors were stably expressed in HEK293 cells and co-purified with cytoskeletal proteins including actin. Disruption of the actin cytoskeleton with cytochalasin D inhibited P2X1 receptor currents with no effect on the time course of the response or surface expression of the receptor. Stabilization of the cytoskeleton with jasplakinolide had no effect on P2X1 receptor currents but decreased receptor mobility. P2X2 receptor currents were unaffected by cytochalasin, and P2X1/2 receptor chimeras were used to identify the molecular basis of actin sensitivity. These studies showed that the intracellular amino terminus accounts for the inhibitory effects of cytoskeletal disruption similar to that shown for lipid raft/cholesterol sensitivity. Stabilization of the cytoskeleton with jasplakinolide abolished the inhibitory effects of cholesterol depletion on P2X1 receptor currents, suggesting that lipid rafts may regulate the receptor through stabilization of the cytoskeleton. These studies show that the cytoskeleton plays an important role in P2X1 receptor regulation. PMID:21757694

TaER Expression Is Associated with Transpiration Efficiency Traits and Yield in Bread Wheat

PubMed Central

Zheng, Jiacheng; Yang, Zhiyuan; Madgwick, Pippa J.; Carmo-Silva, Elizabete; Parry, Martin A. J.; Hu, Yin-Gang

2015-01-01

ERECTA encodes a receptor-like kinase and is proposed as a candidate for determining transpiration efficiency of plants. Two genes homologous to ERECTA in Arabidopsis were identified on chromosomes 6 (TaER2) and 7 (TaER1) of bread wheat (Triticum aestivum L.), with copies of each gene on the A, B and D genomes of wheat. Similar expression patterns were observed for TaER1 and TaER2 with relatively higher expression of TaER1 in flag leaves of wheat at heading (Z55) and grain-filling (Z73) stages. Significant variations were found in the expression levels of both TaER1 and TaER2 in the flag leaves at both growth stages among 48 diverse bread wheat varieties. Based on the expression of TaER1 and TaER2, the 48 wheat varieties could be classified into three groups having high (5 varieties), medium (27 varieties) and low (16 varieties) levels of TaER expression. Significant differences were also observed between the three groups varying for TaER expression for several transpiration efficiency (TE)- related traits, including stomatal density (SD), transpiration rate, photosynthetic rate (A), instant water use efficiency (WUEi) and carbon isotope discrimination (CID), and yield traits of biomass production plant-1 (BYPP) and grain yield plant-1 (GYPP). Correlation analysis revealed that the expression of TaER1 and TaER2 at the two growth stages was significantly and negatively associated with SD (P<0.01), transpiration rate (P<0.05) and CID (P<0.01), while significantly and positively correlated with flag leaf area (FLA, P<0.01), A (P<0.05), WUEi (P<0.05), BYPP (P<0.01) and GYPP (P<0.01), with stronger correlations for TaER1 than TaER2 and at grain-filling stage than at heading stage. These combined results suggested that TaER involved in development of transpiration efficiency -related traits and yield in bread wheat, implying a function for TaER in regulating leaf development of bread wheat and contributing to expression of these traits. Moreover, the results indicate that TaER could be exploitable for manipulating important agronomical traits in wheat improvement. PMID:26047019

TaER Expression Is Associated with Transpiration Efficiency Traits and Yield in Bread Wheat.

PubMed

Zheng, Jiacheng; Yang, Zhiyuan; Madgwick, Pippa J; Carmo-Silva, Elizabete; Parry, Martin A J; Hu, Yin-Gang

2015-01-01

ERECTA encodes a receptor-like kinase and is proposed as a candidate for determining transpiration efficiency of plants. Two genes homologous to ERECTA in Arabidopsis were identified on chromosomes 6 (TaER2) and 7 (TaER1) of bread wheat (Triticum aestivum L.), with copies of each gene on the A, B and D genomes of wheat. Similar expression patterns were observed for TaER1 and TaER2 with relatively higher expression of TaER1 in flag leaves of wheat at heading (Z55) and grain-filling (Z73) stages. Significant variations were found in the expression levels of both TaER1 and TaER2 in the flag leaves at both growth stages among 48 diverse bread wheat varieties. Based on the expression of TaER1 and TaER2, the 48 wheat varieties could be classified into three groups having high (5 varieties), medium (27 varieties) and low (16 varieties) levels of TaER expression. Significant differences were also observed between the three groups varying for TaER expression for several transpiration efficiency (TE)- related traits, including stomatal density (SD), transpiration rate, photosynthetic rate (A), instant water use efficiency (WUEi) and carbon isotope discrimination (CID), and yield traits of biomass production plant-1 (BYPP) and grain yield plant-1 (GYPP). Correlation analysis revealed that the expression of TaER1 and TaER2 at the two growth stages was significantly and negatively associated with SD (P<0.01), transpiration rate (P<0.05) and CID (P<0.01), while significantly and positively correlated with flag leaf area (FLA, P<0.01), A (P<0.05), WUEi (P<0.05), BYPP (P<0.01) and GYPP (P<0.01), with stronger correlations for TaER1 than TaER2 and at grain-filling stage than at heading stage. These combined results suggested that TaER involved in development of transpiration efficiency -related traits and yield in bread wheat, implying a function for TaER in regulating leaf development of bread wheat and contributing to expression of these traits. Moreover, the results indicate that TaER could be exploitable for manipulating important agronomical traits in wheat improvement.

Symbolic Speech

ERIC Educational Resources Information Center

Podgor, Ellen S.

1976-01-01

The concept of symbolic speech emanates from the 1967 case of United States v. O'Brien. These discussions of flag desecration, grooming and dress codes, nude entertainment, buttons and badges, and musical expression show that the courts place symbolic speech in different strata from verbal communication. (LBH)

Mouse Na+/K+-ATPase β1-subunit has a K+-dependent cell adhesion activity for β-GlcNAc-terminating glycans

PubMed Central

Kitamura, Noriaki; Ikekita, Masahiko; Sato, Takeshi; Akimoto, Yoshihiro; Hatanaka, Yasumaru; Kawakami, Hayato; Inomata, Mitsushi; Furukawa, Kiyoshi

2005-01-01

A 48-kDa β-N-acetylglucosamine (GlcNAc)-binding protein was isolated from mouse brain by GlcNAc-agarose column chromatography. The N-terminal amino acid residues showed the protein to be a mouse Na+/K+-ATPase β1-subunit. When the recombinant FLAG-β1-subunit expressed in Sf-9 cells was applied to a GlcNAc-agarose column, only the glycosylated 38- and 40-kDa proteins bound to the column. In the absence of KCl, little of the proteins bound to a GlcNAc-agarose column, but the 38- and 40-kDa proteins bound in the presence of KCl at concentrations above 1 mM. Immunohistochemical study showed that the β1-subunit and GlcNAc-terminating oligosaccharides are at the cell contact sites. Inclusion of anti-β1-subunit antibody or chitobiose in cell aggregation assays using mouse neural cells resulted in inhibition of cell aggregation. These results indicate that the Na+/K+-ATPase β1-subunit is a potassium-dependent lectin that binds to GlcNAc-terminating oligosaccharides: it may be involved in neural cell interactions. PMID:15705719

Scalability in Production System Programs

DTIC Science & Technology

1994-01-01

met this tall, stooping, mostly bald man who glared at me through large glasses . I guess I must have liked being glared at because within a few days, he...yes) < ceill >) {(cell Aid <id> ^modified no) <cell2>) (modify <celll> ^modified no) (remove <ce12>)) (p stage-computation-O (flag compaite-cells

A prognostic model for survival after salvage treatment with FLAG-Ida +/- gemtuzumab-ozogamicine in adult patients with refractory/relapsed acute myeloid leukaemia.

PubMed

Bergua, Juan M; Montesinos, Pau; Martinez-Cuadrón, David; Fernández-Abellán, Pascual; Serrano, Josefina; Sayas, María J; Prieto-Fernandez, Julio; García, Raimundo; García-Huerta, Ana J; Barrios, Manuel; Benavente, Celina; Pérez-Encinas, Manuel; Simiele, Adriana; Rodríguez-Macias, Gabriela; Herrera-Puente, Pilar; Rodríguez-Veiga, Rebeca; Martínez-Sánchez, María P; Amador-Barciela, María L; Riaza-Grau, Rosalía; Sanz, Miguel A

2016-09-01

The combination of fludarabine, cytarabine, idarubicin, and granulocyte colony-stimulating factor (FLAG-Ida) is widely used in relapsed/refractory acute myeloid leukaemia (AML). We retrospectively analysed the results of 259 adult AML patients treated as first salvage with FLAG-Ida or FLAG-Ida plus Gentuzumab-Ozogamicin (FLAGO-Ida) of the Programa Español de Tratamientos en Hematología (PETHEMA) database, developing a prognostic score system of survival in this setting (SALFLAGE score). Overall, 221 patients received FLAG-Ida and 38 FLAGO-Ida; 92 were older than 60 years. The complete remission (CR)/CR with incomplete blood count recovery (CRi) rate was 51%, with 9% of induction deaths. Three covariates were associated with lower CR/CRi: high-risk cytogenetics and t(8;21) at diagnosis, no previous allogeneic stem cell transplantation (allo-SCT) and relapse-free interval <1 year. Allo-SCT was performed in second CR in 60 patients (23%). The median overall survival (OS) of the entire cohort was 0·7 years, with 22% OS at 5-years. Four independent variables were used to construct the score: cytogenetics, FLT3-internal tandem duplication, length of relapse-free interval and previous allo-SCT. Using this stratification system, three groups were defined: favourable (26% of patients), intermediate (29%) and poor-risk (45%), with an expected 5-year OS of 52%, 26% and 7%, respectively. The SALFLAGE score discriminated a subset of patients with an acceptable long-term outcome using FLAG-Ida/FLAGO-Ida regimen. The results of this retrospective analysis should be validated in independent external cohorts. © 2016 John Wiley & Sons Ltd.

The cannabinoid receptor CB1 modulates the signaling properties of the lysophosphatidylinositol receptor GPR55.

PubMed

Kargl, Julia; Balenga, Nariman; Parzmair, Gerald P; Brown, Andrew J; Heinemann, Akos; Waldhoer, Maria

2012-12-28

The G protein-coupled receptor (GPCR) 55 (GPR55) and the cannabinoid receptor 1 (CB1R) are co-expressed in many tissues, predominantly in the central nervous system. Seven transmembrane spanning (7TM) receptors/GPCRs can form homo- and heteromers and initiate distinct signaling pathways. Recently, several synthetic CB1 receptor inverse agonists/antagonists, such as SR141716A, AM251, and AM281, were reported to activate GPR55. Of these, SR141716A was marketed as a promising anti-obesity drug, but was withdrawn from the market because of severe side effects. Here, we tested whether GPR55 and CB1 receptors are capable of (i) forming heteromers and (ii) whether such heteromers could exhibit novel signaling patterns. We show that GPR55 and CB1 receptors alter each others signaling properties in human embryonic kidney (HEK293) cells. We demonstrate that the co-expression of FLAG-CB1 receptors in cells stably expressing HA-GPR55 specifically inhibits GPR55-mediated transcription factor activation, such as nuclear factor of activated T-cells and serum response element, as well as extracellular signal-regulated kinases (ERK1/2) activation. GPR55 and CB1 receptors can form heteromers, but the internalization of both receptors is not affected. In addition, we observe that the presence of GPR55 enhances CB1R-mediated ERK1/2 and nuclear factor of activated T-cell activation. Our data provide the first evidence that GPR55 can form heteromers with another 7TM/GPCR and that this interaction with the CB1 receptor has functional consequences in vitro. The GPR55-CB1R heteromer may play an important physiological and/or pathophysiological role in tissues endogenously co-expressing both receptors.

Brief Report: A mass spectrometry assay to simultaneously analyze ROS1 and RET fusion gene expression in non-small cell lung cancer

PubMed Central

Wijesinghe, Priyanga; Bepler, Gerold

2014-01-01

Introduction ROS1 and RET gene fusions were recently discovered in non-small cell lung cancer (NSCLC) as potential therapeutic targets with small molecule kinase inhibitors. The conventional screening methods of these fusions are time consuming and require samples of high quality and quantity. Here, we describe a novel and efficient method by coupling the power of multiplexing PCR and the sensitivity of mass spectrometry. Methods The multiplex mass spectrometry platform simultaneously tests samples for the expression of nine ROS1 and six RET fusion genes. The assay incorporates detection of wild-type exon junctions immediately upstream and downstream of the fusion junction to exclude false negative results. To flag false positives, the system also comprises two independent assays for each fusion gene junction. Results The characteristic mass spectrometric peaks of the gene fusions were obtained using engineered plasmid constructs. Specific assays targeting the wild-type gene exon junctions were validated using cDNA from lung tissue of healthy individuals. The system was further validated using cDNA derived from NSCLC cell lines that express endogenous fusion genes. The expressed ROS1-SLC34A2 and CCDC6-RET gene fusions from the NSCLC cell lines HCC78 and LC-2/ad, respectively, were accurately detected by the novel assay. The assay is extremely sensitive, capable of detecting an event in test specimens containing 0.5% positive tumors. Conclusion The novel multiplexed assay is robustly capable of detecting 15 different clinically relevant RET and ROS1 fusion variants. The benefits of this detection method include exceptionally low sample input, high cost efficiency, flexibility, and rapid turnover. PMID:25384172

The Cannabinoid Receptor CB1 Modulates the Signaling Properties of the Lysophosphatidylinositol Receptor GPR55*

PubMed Central

Kargl, Julia; Balenga, Nariman; Parzmair, Gerald P.; Brown, Andrew J.; Heinemann, Akos; Waldhoer, Maria

2012-01-01

The G protein-coupled receptor (GPCR) 55 (GPR55) and the cannabinoid receptor 1 (CB1R) are co-expressed in many tissues, predominantly in the central nervous system. Seven transmembrane spanning (7TM) receptors/GPCRs can form homo- and heteromers and initiate distinct signaling pathways. Recently, several synthetic CB1 receptor inverse agonists/antagonists, such as SR141716A, AM251, and AM281, were reported to activate GPR55. Of these, SR141716A was marketed as a promising anti-obesity drug, but was withdrawn from the market because of severe side effects. Here, we tested whether GPR55 and CB1 receptors are capable of (i) forming heteromers and (ii) whether such heteromers could exhibit novel signaling patterns. We show that GPR55 and CB1 receptors alter each others signaling properties in human embryonic kidney (HEK293) cells. We demonstrate that the co-expression of FLAG-CB1 receptors in cells stably expressing HA-GPR55 specifically inhibits GPR55-mediated transcription factor activation, such as nuclear factor of activated T-cells and serum response element, as well as extracellular signal-regulated kinases (ERK1/2) activation. GPR55 and CB1 receptors can form heteromers, but the internalization of both receptors is not affected. In addition, we observe that the presence of GPR55 enhances CB1R-mediated ERK1/2 and nuclear factor of activated T-cell activation. Our data provide the first evidence that GPR55 can form heteromers with another 7TM/GPCR and that this interaction with the CB1 receptor has functional consequences in vitro. The GPR55-CB1R heteromer may play an important physiological and/or pathophysiological role in tissues endogenously co-expressing both receptors. PMID:23161546

On Fock-space representations of quantized enveloping algebras related to noncommutative differential geometry

NASA Astrophysics Data System (ADS)

Jurčo, B.; Schlieker, M.

1995-07-01

In this paper explicitly natural (from the geometrical point of view) Fock-space representations (contragradient Verma modules) of the quantized enveloping algebras are constructed. In order to do so, one starts from the Gauss decomposition of the quantum group and introduces the differential operators on the corresponding q-deformed flag manifold (assumed as a left comodule for the quantum group) by a projection to it of the right action of the quantized enveloping algebra on the quantum group. Finally, the representatives of the elements of the quantized enveloping algebra corresponding to the left-invariant vector fields on the quantum group are expressed as first-order differential operators on the q-deformed flag manifold.

Identification of transactivation-responsive DNA-binding protein 43 (TARDBP43; TDP-43) as a novel factor for TNF-α expression upon lipopolysaccharide stimulation in human monocytes.

PubMed

Murata, H; Hattori, T; Maeda, H; Takashiba, S; Takigawa, M; Kido, J; Nagata, T

2015-08-01

Tumor necrosis factor alpha (TNF-α) is a major cytokine implicated in various inflammatory diseases. The nature of the nuclear factors associated with human TNF-α gene regulation is not well elucidated. We previously identified a novel region located from -550 to -487 in human TNF-α promoter that did not contain the reported binding sites for nuclear factor kappa B (NF-κB) but showed lipopolysaccharide (LPS)-induced transcriptional activity. The purpose of this study is to identify novel factors that bind to the promoter region and regulate TNF-α expression. To identify DNA-binding proteins that bound to the target region of TNF-α promoter, a cDNA library from LPS-stimulated human monocytic cell line THP-1 was screened using a yeast one-hybrid system. Cellular localizations of the DNA-binding protein in the cells were examined by subcellular immunocytochemistry. Nuclear amounts of the protein in LPS-stimulated THP-1 cells were identified by western blot analysis. Expression of mRNA of the protein in the cells was quantified by real-time polymerase chain reaction. Electrophoretic mobility shift assays were performed to confirm the DNA-binding profile. Overexpression of the protein and knockdown of the gene were also performed to investigate the role for TNF-α expression. Several candidates were identified from the cDNA library and transactivation-responsive DNA-binding protein 43 (TARDBP43; TDP-43) was focused on. Western blot analysis revealed that nuclear TDP-43 protein was increased in the LPS-stimulated THP-1 cells. Expression of TDP-43 mRNA was already enhanced before TNF-α induction by LPS. Electrophoretic mobility shift assay analysis showed that nuclear extracts obtained by overexpressing FLAG-tagged TDP-43 bound to the -550 to -487 TNF-α promoter fragments. Overexpression of TDP-43 in THP-1 cells resulted in an increase of TNF-α expression. Knockdown of TDP-43 in THP-1 cells downregulated TNF-α expression. We identified TDP-43 as one of the novel TNF-α factors and found that it bound to the LPS-responsive element in the TNF-α promoter to increase TNF-α expression. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

10 CFR 1002.21 - Description of distinguishing flag.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 10 Energy 4 2010-01-01 2010-01-01 false Description of distinguishing flag. 1002.21 Section 1002.21 Energy DEPARTMENT OF ENERGY (GENERAL PROVISIONS) OFFICIAL SEAL AND DISTINGUISHING FLAG Distinguishing Flag § 1002.21 Description of distinguishing flag. (a) The base or field of the flag shall be...

The long N-terminus of the human monocarboxylate transporter 8 is a target of ubiquitin-dependent proteasomal degradation which regulates protein expression and oligomerization capacity.

PubMed

Zwanziger, Denise; Schmidt, Mathias; Fischer, Jana; Kleinau, Gunnar; Braun, Doreen; Schweizer, Ulrich; Moeller, Lars Christian; Biebermann, Heike; Fuehrer, Dagmar

2016-10-15

Monocarboxylate transporter 8 (MCT8) equilibrates thyroid hormones between the extra- and the intracellular sides. MCT8 exists either with a short or a long N-terminus, but potential functional differences between both variants are yet not known. We, therefore, generated MCT8 constructs which are different in N-terminal length: MCT8(1-613), MCT8(25-613), MCT8(49-613) and MCT8(75-613). The M75G substitution prevents translation of MCT8(75-613) and ensures expression of full-length MCT8 protein. The K56G substitution was made to prevent ubiquitinylation. Cell-surface expression, localization and proteasomal degradation were investigated using C-terminally GFP-tagged MCT8 constructs (HEK293 and MDCK1 cells) and oligomerization capacity was determined using N-terminally HA- and C-terminally FLAG-tagged MCT8 constructs (COS7 cells). MCT8(1-613)-GFP showed a lower protein expression than the shorter MCT8(75-613)-GFP protein. The proteasome inhibitor lactacystin increased MCT8(1-613)-GFP protein amount, suggesting proteasomal degradation of MCT8 with the long N-terminus. Ubiquitin conjugation of MCT8(1-613)-GFP was found by immuno-precipitation. A diminished ubiquitin conjugation caused by K56G substitution resulted in increased MCT8(1-613)-GFP protein expression. Sandwich ELISA was performed to investigate if the bands at higher molecular weight observed in Western blot analysis are due to MCT8 oligomerization, which was indeed shown. Our data imply a role of the long N-terminus of MCT8 as target of ubiquitin-dependent proteasomal degradation affecting MCT8 amount and subsequently oligomerization capacity. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

10 CFR 1002.22 - Use of distinguishing flag.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 10 Energy 4 2010-01-01 2010-01-01 false Use of distinguishing flag. 1002.22 Section 1002.22 Energy DEPARTMENT OF ENERGY (GENERAL PROVISIONS) OFFICIAL SEAL AND DISTINGUISHING FLAG Distinguishing Flag § 1002.22 Use of distinguishing flag. (a) DOE distinguishing flags may be used only: (1) In the offices of the...

OCT4B1 Regulates the Cellular Stress Response of Human Dental Pulp Cells with Inflammation

PubMed Central

Liu, Lu; Huang, Rong; Yang, Ruiqi

2017-01-01

Introduction. Infection and apoptosis are combined triggers for inflammation in dental tissues. Octamer-binding transcription factor 4-B1 (OCT4B1), a novel spliced variant of OCT4 family, could respond to the cellular stress and possess antiapoptotic property. However, its specific role in dental pulpitis remains unknown. Methods. To investigate the effect of OCT4B1 on inflammation of dental pulp cells (DPCs), its expression in inflamed dental pulp tissues and DPCs was examined by in situ hybridization, real-time PCR, and FISH assay. OCT4B1 overexpressed DPCs model was established, confirmed by western blot and immunofluorescence staining, and then stimulated with Lipopolysaccharide (LPS). Apoptotic rate was determined by Hoechst/PI staining and FACS. Cell survival rate was calculated by CCK8 assay. Results. In situ hybridization, real-time PCR, and FISH assay revealed that OCT4B1 was extensively expressed in inflamed dental pulp tissues and DPCs with LPS stimulation. Western blot and immunofluorescence staining showed the expression of OCT4B1 and OCT4B increased after OCT4B1 transfection. Hoechst/PI staining and FACS demonstrated that less red/blue fluorescence was detected and apoptotic percentage decreased (3.45%) after transfection. CCK8 demonstrated that the survival rate of pCDH-OCT4B1-flag cells increased. Conclusions. OCT4B1 plays an essential role in inflammation and apoptosis of DPCs. OCT4B might operate synergistically with OCT4B1 to reduce apoptosis. PMID:28473980

A novel method for purification of the endogenously expressed fission yeast Set2 complex.

PubMed

Suzuki, Shota; Nagao, Koji; Obuse, Chikashi; Murakami, Yota; Takahata, Shinya

2014-05-01

Chromatin-associated proteins are heterogeneously and dynamically composed. To gain a complete understanding of DNA packaging and basic nuclear functions, it is important to generate a comprehensive inventory of these proteins. However, biochemical purification of chromatin-associated proteins is difficult and is accompanied by concerns over complex stability, protein solubility and yield. Here, we describe a new method for optimized purification of the endogenously expressed fission yeast Set2 complex, histone H3K36 methyltransferase. Using the standard centrifugation procedure for purification, approximately half of the Set2 protein separated into the insoluble chromatin pellet fraction, making it impossible to recover the large amounts of soluble Set2. To overcome this poor recovery, we developed a novel protein purification technique termed the filtration/immunoaffinity purification/mass spectrometry (FIM) method, which eliminates the need for centrifugation. Using the FIM method, in which whole cell lysates were filtered consecutively through eight different pore sizes (53-0.8μm), a high yield of soluble FLAG-tagged Set2 was obtained from fission yeast. The technique was suitable for affinity purification and produced a low background. A mass spectrometry analysis of anti-FLAG immunoprecipitated proteins revealed that Rpb1, Rpb2 and Rpb3, which have all been reported previously as components of the budding yeast Set2 complex, were isolated from fission yeast using the FIM method. In addition, other subunits of RNA polymerase II and its phosphatase were also identified. In conclusion, the FIM method is valid for the efficient purification of protein complexes that separate into the insoluble chromatin pellet fraction during centrifugation. Copyright © 2014 Elsevier Inc. All rights reserved.

Human Mammary Epithelial Cell Transformation by Rho GTPase Through a Novel Mechanism

DTIC Science & Technology

2009-08-01

cells were transfected with myc-tagged Prks -ROCK1 or Prks -ROCK2, flag-tagged Prc-PKN-AL, or mDia1 using calcium phosphate method. The transfectants...plasmids, Prks -ROCK1, Prks -ROCK2, Prc-PKN-AL, and pFL-mDia1, were transfected into 293T cells, and cell lysates were incubated with GTP-g-S–loaded GST

The induction of H3K9 methylation by PIWIL4 at the p16{sup Ink4a} locus

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

Sugimoto, Keiki; Kage, Hidenori; Aki, Naomi

The field of epigenetics has made progress by the identification of the small RNA-mediated epigenetic modification. However, little is known about the key proteins. Here, we report that the human PIWI-like family is a candidate protein that is involved in the pathway responsible for chromatin remodeling. The PIWI-like family proteins, expressed as the Flag-fusion proteins, formed a bulky body and localized to the nuclear periphery. Transient transfection of PIWI-like 4 (PIWIL4), only member of the PIWI-like family that was ubiquitously expressed in human tissues, induced histone H3 lysine 9 methylation at the p16{sup Ink4a} (CDKN2A) locus. The elevated level ofmore » histone methylation resulted in the downregulation of the p16{sup Ink4a} gene. These results suggest PIWIL4 plays important roles in the chromatin-modifying pathway in human somatic cells.« less

Involvement of NADPH oxidase isoforms in the production of O2− manipulated by ABA in the senescing leaves of early-senescence-leaf (esl) mutant rice (Oryza sativa)

PubMed Central

Wang, Fubiao; Zhao, Qian; Liu, Jianchao; Cheng, Fangmin

2018-01-01

In this study, the differences in reactive oxygen species (ROS) generation and abscisic acid (ABA) accumulation in senescing leaves were investigated by early-senescence-leaf (esl) mutant and its wild type, to clarify the relationship among ABA levels, ROS generation, and NADPH oxidase (Nox) in senescing leaves of rice (Oryza sativa). The temporal expression levels of OsNox isoforms in senescing leaves and their expression patterns in response to ABA treatment were determined through quantitative real-time reverse transcription PCR (qRT-PCR). Results showed that the flag leaf of the esl mutant generated more O2- concentrations and accumulated higher ABA levels than the wild-type cultivar did in the grain-filling stage. Exogenous ABA treatment induced O2- generation; however, it was depressed by diphenyleneiodonium chloride (DPI) pretreatment in the detached leaf segments. This finding suggested the involvement of NADPH oxidase in ABA-induced O2- generation. The esl mutant exhibited significantly higher expression of OsNox2, OsNox5, OsNox6, and OsNox7 in the initial of grain-filling stage, followed by sharply decrease. The transcriptional levels of OsNox1, OsNox3, and OsFR07 in the flag leaf of the esl mutant were significantly lower than those in the wild-type cultivar. The expression levels of OsNox2, OsNox5, OsNox6, and OsNox7 were significantly enhanced by exogenous ABA treatments. The enhanced expression levels of OsNox2 and OsNox6 were dependent on the duration of ABA treatment. The inducible expression levels of OsNox5 and OsNox7 were dependent on ABA concentrations. By contrast, exogenous ABA treatment severely repressed the transcripts of OsNox1, OsNox3, and OsFR07 in the detached leaf segments. Therefore, OsNox2, OsNox5, OsNox6, and OsNox7 were probably involved in the ABA-induced O2- generation in the initial stage of leaf senescence. Subsequently, other oxidases activated in deteriorating cells were associated with ROS generation and accumulation in the senescing leaves of the esl mutant. Conversely, OsNox1, OsNox3, and OsFR07 were not associated with ABA-induced O2- generation during leaf senescence. PMID:29309410

Involvement of NADPH oxidase isoforms in the production of O2- manipulated by ABA in the senescing leaves of early-senescence-leaf (esl) mutant rice (Oryza sativa).

PubMed

Li, Zhaowei; Wang, Fubiao; Zhao, Qian; Liu, Jianchao; Cheng, Fangmin

2018-01-01

In this study, the differences in reactive oxygen species (ROS) generation and abscisic acid (ABA) accumulation in senescing leaves were investigated by early-senescence-leaf (esl) mutant and its wild type, to clarify the relationship among ABA levels, ROS generation, and NADPH oxidase (Nox) in senescing leaves of rice (Oryza sativa). The temporal expression levels of OsNox isoforms in senescing leaves and their expression patterns in response to ABA treatment were determined through quantitative real-time reverse transcription PCR (qRT-PCR). Results showed that the flag leaf of the esl mutant generated more O2- concentrations and accumulated higher ABA levels than the wild-type cultivar did in the grain-filling stage. Exogenous ABA treatment induced O2- generation; however, it was depressed by diphenyleneiodonium chloride (DPI) pretreatment in the detached leaf segments. This finding suggested the involvement of NADPH oxidase in ABA-induced O2- generation. The esl mutant exhibited significantly higher expression of OsNox2, OsNox5, OsNox6, and OsNox7 in the initial of grain-filling stage, followed by sharply decrease. The transcriptional levels of OsNox1, OsNox3, and OsFR07 in the flag leaf of the esl mutant were significantly lower than those in the wild-type cultivar. The expression levels of OsNox2, OsNox5, OsNox6, and OsNox7 were significantly enhanced by exogenous ABA treatments. The enhanced expression levels of OsNox2 and OsNox6 were dependent on the duration of ABA treatment. The inducible expression levels of OsNox5 and OsNox7 were dependent on ABA concentrations. By contrast, exogenous ABA treatment severely repressed the transcripts of OsNox1, OsNox3, and OsFR07 in the detached leaf segments. Therefore, OsNox2, OsNox5, OsNox6, and OsNox7 were probably involved in the ABA-induced O2- generation in the initial stage of leaf senescence. Subsequently, other oxidases activated in deteriorating cells were associated with ROS generation and accumulation in the senescing leaves of the esl mutant. Conversely, OsNox1, OsNox3, and OsFR07 were not associated with ABA-induced O2- generation during leaf senescence.

Dual role of the carboxyl-terminal region of pig liver L-kynurenine 3-monooxygenase: mitochondrial-targeting signal and enzymatic activity.

PubMed

Hirai, Kumiko; Kuroyanagi, Hidehito; Tatebayashi, Yoshitaka; Hayashi, Yoshitaka; Hirabayashi-Takahashi, Kanako; Saito, Kuniaki; Haga, Seiich; Uemura, Tomihiko; Izumi, Susumu

2010-12-01

l-kynurenine 3-monooxygenase (KMO) is an NAD(P)H-dependent flavin monooxygenase that catalyses the hydroxylation of l-kynurenine to 3-hydroxykynurenine, and is localized as an oligomer in the mitochondrial outer membrane. In the human brain, KMO may play an important role in the formation of two neurotoxins, 3-hydroxykynurenine and quinolinic acid, both of which provoke severe neurodegenerative diseases. In mosquitos, it plays a role in the formation both of eye pigment and of an exflagellation-inducing factor (xanthurenic acid). Here, we present evidence that the C-terminal region of pig liver KMO plays a dual role. First, it is required for the enzymatic activity. Second, it functions as a mitochondrial targeting signal as seen in monoamine oxidase B (MAO B) or outer membrane cytochrome b(5). The first role was shown by the comparison of the enzymatic activity of two mutants (C-terminally FLAG-tagged KMO and carboxyl-terminal truncation form, KMOΔC50) with that of the wild-type enzyme expressed in COS-7 cells. The second role was demonstrated with fluorescence microscopy by the comparison of the intracellular localization of the wild-type, three carboxyl-terminal truncated forms (ΔC20, ΔC30 and ΔC50), C-terminally FLAG-tagged wild-type and a mutant KMO, where two arginine residues, Arg461-Arg462, were replaced with Ser residues.

Activity and distribution of intracellular carbonic anhydrase II and their effects on the transport activity of anion exchanger AE1/SLC4A1.

PubMed

Al-Samir, Samer; Papadopoulos, Symeon; Scheibe, Renate J; Meißner, Joachim D; Cartron, Jean-Pierre; Sly, William S; Alper, Seth L; Gros, Gerolf; Endeward, Volker

2013-10-15

We have investigated the previously published 'metabolon hypothesis' postulating that a close association of the anion exchanger 1 (AE1) and cytosolic carbonic anhydrase II (CAII) exists that greatly increases the transport activity of AE1. We study whether there is a physical association of and direct functional interaction between CAII and AE1 in the native human red cell and in tsA201 cells coexpressing heterologous fluorescent fusion proteins CAII-CyPet and YPet-AE1. In these doubly transfected tsA201 cells, YPet-AE1 is clearly associated with the cell membrane, whereas CAII-CyPet is homogeneously distributed throughout the cell in a cytoplasmic pattern. Förster resonance energy transfer measurements fail to detect close proximity of YPet-AE1 and CAII-CyPet. The absence of an association of AE1 and CAII is supported by immunoprecipitation experiments using Flag-antibody against Flag-tagged AE1 expressed in tsA201 cells, which does not co-precipitate native CAII but co-precipitates coexpressed ankyrin. Both the CAII and the AE1 fusion proteins are fully functional in tsA201 cells as judged by CA activity and by cellular HCO3(-) permeability (P(HCO3(-))) sensitive to inhibition by 4,4-Diisothiocyano-2,2-stilbenedisulfonic acid. Expression of the non-catalytic CAII mutant V143Y leads to a drastic reduction of endogenous CAII and to a corresponding reduction of total intracellular CA activity. Overexpression of an N-terminally truncated CAII lacking the proposed site of interaction with the C-terminal cytoplasmic tail of AE1 substantially increases intracellular CA activity, as does overexpression of wild-type CAII. These variously co-transfected tsA201 cells exhibit a positive correlation between cellular P(HCO3(-)) and intracellular CA activity. The relationship reflects that expected from changes in cytoplasmic CA activity improving substrate supply to or removal from AE1, without requirement for a CAII-AE1 metabolon involving physical interaction. A functional contribution of the hypothesized CAII-AE1 metabolon to erythroid AE1-mediated HCO3(-) transport was further tested in normal red cells and red cells from CAII-deficient patients that retain substantial CA activity associated with the erythroid CAI protein lacking the proposed AE1-binding sequence. Erythroid P(HCO3(-)) was indistinguishable in these two cell types, providing no support for the proposed functional importance of the physical interaction of CAII and AE1. A theoretical model predicts that homogeneous cytoplasmic distribution of CAII is more favourable for cellular transport of HCO3(-) and CO2 than is association of CAII with the cytoplasmic surface of the plasma membrane. This is due to the fact that the relatively slow intracellular transport of H(+) makes it most efficient to place the CA in the vicinity of the haemoglobin molecules, which are homogeneously distributed over the cytoplasm.

Jewelry Design by Example

ERIC Educational Resources Information Center

Mahoney, Ellen

2011-01-01

Presidential campaigns bring out the expression of strong political sentiments in the jewelry and pins that people wear to declare their allegiances. Images, flags, symbols, and words are all elements that are incorporated into designs that make decisive visual statements. In this article, the author's high-school metals class students design a…

The Shirts Off Their Backs.

ERIC Educational Resources Information Center

Dowling-Sendor, Benjamin

2001-01-01

In "Castorina," the Sixth Circuit reversed a lower court ruling that had dismissed a suit by two high school students challenging their suspension for wearing T-shirts adorned with the Confederate flag. Reviews three Supreme Court decisions about the regulation of student expression. Concludes that a consensus for the regulation of…

10 CFR 1002.31 - Unauthorized uses of the seal and flag.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 10 Energy 4 2010-01-01 2010-01-01 false Unauthorized uses of the seal and flag. 1002.31 Section 1002.31 Energy DEPARTMENT OF ENERGY (GENERAL PROVISIONS) OFFICIAL SEAL AND DISTINGUISHING FLAG Unauthorized Uses § 1002.31 Unauthorized uses of the seal and flag. The official seal and distinguishing flag...

10 CFR 1002.31 - Unauthorized uses of the seal and flag.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 10 Energy 4 2011-01-01 2011-01-01 false Unauthorized uses of the seal and flag. 1002.31 Section 1002.31 Energy DEPARTMENT OF ENERGY (GENERAL PROVISIONS) OFFICIAL SEAL AND DISTINGUISHING FLAG Unauthorized Uses § 1002.31 Unauthorized uses of the seal and flag. The official seal and distinguishing flag...

49 CFR 393.87 - Warning flags on projecting loads.

Code of Federal Regulations, 2012 CFR

2012-10-01

... 49 Transportation 5 2012-10-01 2012-10-01 false Warning flags on projecting loads. 393.87 Section... ACCESSORIES NECESSARY FOR SAFE OPERATION Miscellaneous Parts and Accessories § 393.87 Warning flags on... load marked with red or orange fluorescent warning flags. Each warning flag must be at least 457 mm (18...

49 CFR 393.87 - Warning flags on projecting loads.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 49 Transportation 5 2011-10-01 2011-10-01 false Warning flags on projecting loads. 393.87 Section... ACCESSORIES NECESSARY FOR SAFE OPERATION Miscellaneous Parts and Accessories § 393.87 Warning flags on... load marked with red or orange fluorescent warning flags. Each warning flag must be at least 457 mm (18...

49 CFR 393.87 - Warning flags on projecting loads.

Code of Federal Regulations, 2013 CFR

2013-10-01

... 49 Transportation 5 2013-10-01 2013-10-01 false Warning flags on projecting loads. 393.87 Section... ACCESSORIES NECESSARY FOR SAFE OPERATION Miscellaneous Parts and Accessories § 393.87 Warning flags on... load marked with red or orange fluorescent warning flags. Each warning flag must be at least 457 mm (18...

49 CFR 393.87 - Warning flags on projecting loads.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 49 Transportation 5 2010-10-01 2010-10-01 false Warning flags on projecting loads. 393.87 Section... ACCESSORIES NECESSARY FOR SAFE OPERATION Miscellaneous Parts and Accessories § 393.87 Warning flags on... load marked with red or orange fluorescent warning flags. Each warning flag must be at least 457 mm (18...

49 CFR 393.87 - Warning flags on projecting loads.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 49 Transportation 5 2014-10-01 2014-10-01 false Warning flags on projecting loads. 393.87 Section... ACCESSORIES NECESSARY FOR SAFE OPERATION Miscellaneous Parts and Accessories § 393.87 Warning flags on... load marked with red or orange fluorescent warning flags. Each warning flag must be at least 457 mm (18...

46 CFR 282.11 - Ranking of flags.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 46 Shipping 8 2010-10-01 2010-10-01 false Ranking of flags. 282.11 Section 282.11 Shipping... COMMERCE OF THE UNITED STATES Foreign-Flag Competition § 282.11 Ranking of flags. The operators under each... priority of costs which are representative of the flag. For liner cargo vessels, the ranking of operators...

Red Flags for Low Back Pain Are Not Always Really Red: A Prospective Evaluation of the Clinical Utility of Commonly Used Screening Questions for Low Back Pain.

PubMed

Premkumar, Ajay; Godfrey, William; Gottschalk, Michael B; Boden, Scott D

2018-03-07

Low back pain has a high prevalence and morbidity, and is a source of substantial health-care spending. Numerous published guidelines support the use of so-called red flag questions to screen for serious pathology in patients with low back pain. This paper examines the effectiveness of red flag questions as a screening tool for patients presenting with low back pain to a multidisciplinary academic spine center. We conducted a retrospective review of the cases of 9,940 patients with a chief complaint of low back pain. The patients completed a questionnaire that included several red flag questions during their first physician visit. Diagnostic data for the same clinical episode were collected from medical records and were corroborated with imaging reports. Patients who were diagnosed as having a vertebral fracture, malignancy, infection, or cauda equina syndrome were classified as having a red flag diagnosis. Specific individual red flags and combinations of red flags were associated with an increased probability of underlying serious spinal pathology, e.g., recent trauma and an age of >50 years were associated with vertebral fracture. The presence or absence of other red flags, such as night pain, was unrelated to any particular diagnosis. For instance, for patients with no recent history of infection and no fever, chills, or sweating, the presence of night pain was a false-positive finding for infection >96% of the time. In general, the absence of red flag responses did not meaningfully decrease the likelihood of a red flag diagnosis; 64% of patients with spinal malignancy had no associated red flags. While a positive response to a red flag question may indicate the presence of serious disease, a negative response to 1 or 2 red flag questions does not meaningfully decrease the likelihood of a red flag diagnosis. Clinicians should use caution when utilizing red flag questions as screening tools.

QTL mapping of flag leaf-related traits in wheat (Triticum aestivum L.).

PubMed

Liu, Kaiye; Xu, Hao; Liu, Gang; Guan, Panfeng; Zhou, Xueyao; Peng, Huiru; Yao, Yingyin; Ni, Zhongfu; Sun, Qixin; Du, Jinkun

2018-04-01

QTL controlling flag leaf length, flag leaf width, flag leaf area and flag leaf angle were mapped in wheat. This study aimed to advance our understanding of the genetic mechanisms underlying morphological traits of the flag leaves of wheat (Triticum aestivum L.). A recombinant inbred line (RIL) population derived from ND3331 and the Tibetan semi-wild wheat Zang1817 was used to identify quantitative trait loci (QTLs) controlling flag leaf length (FLL), flag leaf width (FLW), flag leaf area (FLA), and flag leaf angle (FLANG). Using an available simple sequence repeat genetic linkage map, 23 putative QTLs for FLL, FLW, FLA, and FLANG were detected on chromosomes 1B, 2B, 3A, 3D, 4B, 5A, 6B, 7B, and 7D. Individual QTL explained 4.3-68.52% of the phenotypic variance in different environments. Four QTLs for FLL, two for FLW, four for FLA, and five for FLANG were detected in at least two environments. Positive alleles of 17 QTLs for flag leaf-related traits originated from ND3331 and 6 originated from Zang1817. QTLs with pleiotropic effects or multiple linked QTL were also identified on chromosomes 1B, 4B, and 5A; these are potential target regions for fine-mapping and marker-assisted selection in wheat breeding programs.

AP2 adaptor complex mediates bile salt export pump internalization and modulates its hepatocanalicular expression and transport function.

PubMed

Hayashi, Hisamitsu; Inamura, Kaori; Aida, Kensuke; Naoi, Sotaro; Horikawa, Reiko; Nagasaka, Hironori; Takatani, Tomozumi; Fukushima, Tamio; Hattori, Asami; Yabuki, Takashi; Horii, Ikuo; Sugiyama, Yuichi

2012-06-01

The bile salt export pump (BSEP) mediates the biliary excretion of bile salts and its dysfunction induces intrahepatic cholestasis. Reduced canalicular expression of BSEP resulting from the promotion of its internalization is one of the causes of this disease state. However, the molecular mechanism underlying BSEP internalization from the canalicular membrane (CM) remains unknown. We have shown previously that 4-phenylbutyrate (4PBA), a drug used for ornithine transcarbamylase deficiency (OTCD), inhibited internalization and subsequent degradation of cell-surface-resident BSEP. The current study found that 4PBA treatment decreased significantly the expression of α- and μ2-adaptin, both of which are subunits of the AP2 adaptor complex (AP2) that mediates clathrin-dependent endocytosis, in liver specimens from rats and patients with OTCD, and that BSEP has potential AP2 recognition motifs in its cytosolic region. Based on this, the role of AP2 in BSEP internalization was explored further. In vitro analysis with 3×FLAG-human BSEP-expressing HeLa cells and human sandwich-culture hepatocytes indicates that the impairment of AP2 function by RNA interference targeting of α-adaptin inhibits BSEP internalization from the plasma membrane and increases its cell-surface expression and transport function. Studies using immunostaining, coimmunoprecipitation, glutathione S-transferase pulldown assay, and time-lapse imaging show that AP2 interacts with BSEP at the CM through a tyrosine motif at the carboxyl terminus of BSEP and mediates BSEP internalization from the CM of hepatocytes. AP2 mediates the internalization and subsequent degradation of CM-resident BSEP through direct interaction with BSEP and thereby modulates the canalicular expression and transport function of BSEP. This information should be useful for understanding the pathogenesis of severe liver diseases associated with intrahepatic cholestasis. Copyright © 2012 American Association for the Study of Liver Diseases.

Process to evaluate hematological parameters that reflex to manual differential cell counts in a pediatric institution.

PubMed

Guarner, Jeannette; Atuan, Maria Ana; Nix, Barbara; Mishak, Christopher; Vejjajiva, Connie; Curtis, Cheri; Park, Sunita; Mullins, Richard

2010-01-01

Each institution sets specific parameters obtained by automated hematology analyzers to trigger manual counts. We designed a process to decrease the number of manual differential cell counts without impacting patient care. We selected new criteria that prompt manual counts and studied the impact these changes had in 2 days of work and in samples of patients with newly diagnosed leukemia, sickle cell disease, and presence of left shift. By using fewer parameters and expanding our ranges we decreased the number of manual counts by 20%. The parameters that prompted manual counts most frequently were the presence of blast flags and nucleated red blood cells, 2 parameters that were not changed. The parameters that accounted for a decrease in the number of manual counts were the white blood cell count and large unstained cells. Eight of 32 patients with newly diagnosed leukemia did not show blast flags; however, other parameters triggered manual counts. In 47 patients with sickle cell disease, nucleated red cells and red cell variability prompted manual review. Bands were observed in 18% of the specimens and 4% would not have been counted manually with the new criteria, for the latter the mean band count was 2.6%. The process we followed to evaluate hematological parameters that reflex to manual differential cell counts increased efficiency without compromising patient care in our hospital system.

Comparative in Silico Analysis of Ferric Reduction Oxidase (FRO) Genes Expression Patterns in Response to Abiotic Stresses, Metal and Hormone Applications.

PubMed

Muhammad, Izhar; Jing, Xiu-Qing; Shalmani, Abdullah; Ali, Muhammad; Yi, Shi; Gan, Peng-Fei; Li, Wen-Qiang; Liu, Wen-Ting; Chen, Kun-Ming

2018-05-12

The ferric reduction oxidase (FRO) gene family is involved in various biological processes widely found in plants and may play an essential role in metal homeostasis, tolerance and intricate signaling networks in response to a number of abiotic stresses. Our study describes the identification, characterization and evolutionary relationships of FRO genes families. Here, total 50 FRO genes in Plantae and 15 ‘FRO like’ genes in non-Plantae were retrieved from 16 different species. The entire FRO genes have been divided into seven clades according to close similarity in biological and functional behavior. Three conserved domains were common in FRO genes while in two FROs sub genome have an extra NADPH-Ox domain, separating the function of plant FROs. OsFRO1 and OsFRO7 genes were expressed constitutively in rice plant. Real-time RT-PCR analysis demonstrated that the expression of OsFRO1 was high in flag leaf, and OsFRO7 gene expression was maximum in leaf blade and flag leaf. Both genes showed vigorous expressions level in response to different abiotic and hormones treatments. Moreover, the expression of both genes was also substantial under heavy metal stresses. OsFRO1 gene expression was triggered following 6 h under Zn, Pb, Co and Ni treatments, whereas OsFRO7 gene expression under Fe, Pb and Ni after 12 h, Zn and Cr after 6 h, and Mn and Co after 3 h treatments. These findings suggest the possible involvement of both the genes under abiotic and metal stress and the regulation of phytohormones. Therefore, our current work may provide the foundation for further functional characterization of rice FRO genes family.

77 FR 56896 - Self-Regulatory Organizations; EDGA Exchange, Inc.; Notice of Filing and Immediate Effectiveness...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-09-14

... the relevant flags, as described below, for orders that add liquidity to the EDGA book. Specifically... the following flags: Flag B for orders that add liquidity to the EDGA book in Tape B securities; Flag V for orders that add liquidity to the EDGA book in Tape A securities; Flag Y for orders that add...

FLIP for FLAG model visualization

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

Wooten, Hasani Omar

A graphical user interface has been developed for FLAG users. FLIP (FLAG Input deck Parser) provides users with an organized view of FLAG models and a means for efficiently and easily navigating and editing nodes, parameters, and variables.

Where no flag has gone before: Political and technical aspects of placing a flag on the Moon

NASA Technical Reports Server (NTRS)

Platoff, Anne M.

1993-01-01

The flag on the Moon represents an important event in vexillological history. The political and technical aspects of placing a flag on the Moon, focusing on the first Moon landing, is examined. During their historic extravehicular activity, the Apollo 11 crew planted the flag of the United States on the lunar surface. This flag-raising was strictly a symbolic activity, as the United Nations Treaty on Outer Space precluded any territorial claim. Nevertheless, there were domestic and international debates over the appropriateness of the event. Congress amended the agency's appropriations bill to prevent the National Aeronautics and Space Administration (NASA) from placing flags of other nations, or those of international associations, on the Moon during missions funded solely by the United States. Like any activity in space exploration, the Apollo flag-raising also provided NASA engineers with an interesting technical challenge. They designed a flagpole with a horizontal bar allowing the flag to 'fly' without the benefit of wind to overcome the effects of the Moon's lack of an atmosphere. Other factors considered in the design were weight, heat resistance, and ease of assembly by astronauts whose space suits restricted their range of movement and ability to grasp items. As NASA plans a return to the Moon and an expedition to Mars, we will likely see flags continue to go 'where no flag has gone before'.

Cystatin C–Adiponectin Complex in Plasma Associates with Coronary Plaque Instability

PubMed Central

Matsuoka, Tetsuro; Kayama, Kento; Onishi, Sumire; Matsuo, Natsumi

2017-01-01

Aim: Adiponectin (APN) is an adipocyte-derived bioactive molecule with antiatherogenic properties. We previously reported that cystatin C (CysC) abolished the anti-atherogenic effects of APN. We aimed to elucidate the clinical significance of CysC–APN complex in patients with coronary artery disease (CAD). Methods: We enrolled 43 stable CAD male patients to examine the relationship between CysC–APN complex and coronary plaque characteristics. Serum was immunoprecipitated by the anti-APN antibody and immunoblotted by the anti-CysC antibody to demonstrate the presence of CysC–APN complexes in vivo. To confirm their binding in vitro, HEK293T cell lysates overexpressing myc-APN and FLAG-CysC were immunoprecipitated with an anti-myc or anti-FLAG antibody, followed by immunoblotting with an anti-APN or anti-CysC antibody. Results: CysC was identified as a specific co-immunoprecipitant with APN by the anti-APN antibody in human serum. In vitro, FLAG-CysC was co-immunoprecipitated with myc-APN by the antimyc antibody and myc-APN was co-immunoprecipitated with FLAG-CysC by the anti-FLAG antibody. Among CAD patients, serum CysC–APN complex levels negatively correlated with fibrotic components of coronary plaques and positively correlated with either necrotic or lipidic plus necrotic components. Plaque burden negatively correlated with serum APN levels but not serum CysC–APN complex levels. Serum CysC levels had no association with plaque characteristics. In multivariate analysis, CysC–APN complex levels were identified as the strongest negative factor for fibrotic components and the strongest positive factor for both necrotic and lipidic plus necrotic components. Conclusion: Measuring serum CysC–APN complex levels is helpful for evaluating coronary plaque instability in CAD patients. PMID:28321013

Experimental investigation of flow field around the elastic flag flapping in periodic state

NASA Astrophysics Data System (ADS)

Jia, Yongxia; Jia, Lichao; Su, Zhuang; Yuan, Huijing

2018-05-01

The flapping of a flag in the wind is a classical fluid-structure problem that concerns the interaction of elastic bodies with ambient fluid. We focus on the desirable experimental results of the flow around the flapping flag. By immersing the elastic yet self-supporting heavy flag into water flow, we use particle image velocimetry (PIV) techniques to obtain the whole flow field around the midspan of the flag interacting with a fluid in periodic state. A unique PIV image processing method is used to measure near-wall flow velocities around a moving elastic flag. There exists a thin flow circulation region on the suction side of the flag in periodic state. This observation suggests that viscous flow models may be needed to improve the theoretical predictions of the flapping flag in periodic state, especially in a large amplitude.

Sodium 4-phenylbutyrate ameliorates the effects of cataract-causing mutant gammaD-crystallin in cultured cells.

PubMed

Gong, Bo; Zhang, Li-Yun; Lam, Dennis Shun-Chiu; Pang, Chi-Pui; Yam, Gary Hin-Fai

2010-06-04

gammaD-Crystallin (CRYGD) is a major structural lens crystallin and its mutations result in congenital cataract formation. In this study, we attempted to correct the altered protein features of G165fsX8 CRYGD protein with small chemical molecules. Recombinant FLAG-tagged mutants (R15C, R15S, P24T, R61C, and G165fsX8) of CRYGD were expressed in COS-7 cells and treated with small chemical molecules with reported protein chaperoning properties (sodium 4-phenylbutyrate [4-PBA], trimethylamine N-oxide [TMAO], and glycerol and DMSO [DMSO]). Protein solubility in 0.5% Triton X-100 and subcellular distribution was examined by western blotting and immunofluorescence, respectively. Apoptosis was assayed as the percentage of fragmented nuclei in transfected cells. Expression of heat-shock proteins (Hsp70 and Hsp90) was examined by reverse transcription-polymerase chain reaction analysis. Unlike WT and most mutants (R15C, R15S, P24T, and R61C) of CRYGD, G165fsX8 CRYGD was significantly insoluble in 0.5% Triton X-100. This insolubility was alleviated by dose-dependent 4-PBA treatment. The treatment relieved the mislocalization of G165fsX8 CRYGD from the nuclear envelope. Also, 4-PBA treatment reduced cell apoptosis and caused an upregulation of Hsp70. 4-PBA treatment reduced the defective phenotype of mutant G165fsX8 CRYGD and rescued the affected cells from apoptosis. This could be a potential treatment for lens structural protein and prevent lens opacity in cataract formation.

14 CFR 1221.107 - Establishment of the NASA Administrator's, Deputy Administrator's, and Associate Deputy...

Code of Federal Regulations, 2012 CFR

2012-01-01

... × 4 feet; (2) The Administrator's Flag has four stars; (3) The Deputy Administrator's Flag has three stars; and (4) The Associate Deputy Administrator's Flag has two stars. (b) Flags representing these...

14 CFR § 1221.107 - Establishment of the NASA Administrator's, Deputy Administrator's, and Associate Deputy...

Code of Federal Regulations, 2014 CFR

2014-01-01

... × 4 feet; (2) The Administrator's Flag has four stars; (3) The Deputy Administrator's Flag has three stars; and (4) The Associate Deputy Administrator's Flag has two stars. (b) Flags representing these...

14 CFR 1221.107 - Establishment of the NASA Administrator's, Deputy Administrator's, and Associate Deputy...

Code of Federal Regulations, 2010 CFR

2010-01-01

... × 4 feet; (2) The Administrator's Flag has four stars; (3) The Deputy Administrator's Flag has three stars; and (4) The Associate Deputy Administrator's Flag has two stars. (b) Flags representing these...

14 CFR 1221.107 - Establishment of the NASA Administrator's, Deputy Administrator's, and Associate Deputy...

Code of Federal Regulations, 2013 CFR

2013-01-01

... × 4 feet; (2) The Administrator's Flag has four stars; (3) The Deputy Administrator's Flag has three stars; and (4) The Associate Deputy Administrator's Flag has two stars. (b) Flags representing these...

14 CFR 1221.107 - Establishment of the NASA Administrator's, Deputy Administrator's, and Associate Deputy...

Code of Federal Regulations, 2011 CFR

2011-01-01

... × 4 feet; (2) The Administrator's Flag has four stars; (3) The Deputy Administrator's Flag has three stars; and (4) The Associate Deputy Administrator's Flag has two stars. (b) Flags representing these...

On national flags and language tags: Effects of flag-language congruency in bilingual word recognition.

PubMed

Grainger, Jonathan; Declerck, Mathieu; Marzouki, Yousri

2017-07-01

French-English bilinguals performed a generalized lexical decision experiment with mixed lists of French and English words and pseudo-words. In Experiment 1, each word/pseudo-word was superimposed on the picture of the French or UK flag, and flag-word congruency was manipulated. The flag was not informative with respect to either the lexical decision response or the language of the word. Nevertheless, lexical decisions to word stimuli were faster following the congruent flag compared with the incongruent flag, but only for French (L1) words. Experiment 2 replicated this flag-language congruency effect in a priming paradigm, where the word and pseudo-word targets followed the brief presentation of the flag prime, and this time effects were seen in both languages. We take these findings as evidence for a mechanism that automatically processes linguistic and non-linguistic information concerning the presence or not of a given language. Language membership information can then modulate lexical processing, in line with the architecture of the BIA model, but not the BIA+ model. Copyright © 2017 Elsevier B.V. All rights reserved.

49 CFR 234.5 - Definitions.

Code of Federal Regulations, 2010 CFR

2010-10-01

... devices for daytime flagging include “ STOP/SLOW” paddles or red flags. For nighttime flagging, a... Administration's Manual on Uniform Traffic Control Devices addresses standards and guides for flaggers and... follow them to the greatest extent possible. Copies of the latest MUTCD provisions regarding flagging...

GEC-targeted HO-1 expression reduces proteinuria in glomerular immune injury.

PubMed

Duann, Pu; Lianos, Elias A

2009-09-01

Induction of heme oxygenase (HO)-1 is a key defense mechanism against oxidative stress. Compared with tubules, glomeruli are refractory to HO-1 upregulation in response to injury. This can be a disadvantage as it may be associated with insufficient production of cytoprotective heme-degradation metabolites. We, therefore, explored whether 1) targeted HO-1 expression can be achieved in glomeruli without altering their physiological integrity and 2) this expression reduces proteinuria in immune injury induced by an anti-glomerular basement membrane (GBM) antibody (Ab). We employed a 4.125-kb fragment of a mouse nephrin promoter downstream to which a FLAG-tagged hHO-1 cDNA sequence was inserted and subsequently generated transgenic mice from the FVB/N parental strain. There was a 16-fold higher transgene expression in the kidney than nonspecific background (liver) while the transprotein immunolocalized in glomerular epithelial cells (GEC). There was no change in urinary protein excretion, indicating that GEC-targeted HO-1 expression had no effect on glomerular protein permeability. Urinary protein excretion in transgenic mice with anti-GBM Ab injury (days 3 and 6) was significantly lower compared with wild-type controls. There was no significant change in renal expression levels of profibrotic (TGF-beta1) or anti-inflammatory (IL-10) cytokines in transgenic mice with anti-GBM Ab injury. These observations indicate that GEC-targeted HO-1 expression does not alter glomerular physiological integrity and reduces proteinuria in glomerular immune injury.

GEC-targeted HO-1 expression reduces proteinuria in glomerular immune injury

PubMed Central

Duann, Pu; Lianos, Elias A.

2009-01-01

Induction of heme oxygenase (HO)-1 is a key defense mechanism against oxidative stress. Compared with tubules, glomeruli are refractory to HO-1 upregulation in response to injury. This can be a disadvantage as it may be associated with insufficient production of cytoprotective heme-degradation metabolites. We, therefore, explored whether 1) targeted HO-1 expression can be achieved in glomeruli without altering their physiological integrity and 2) this expression reduces proteinuria in immune injury induced by an anti-glomerular basement membrane (GBM) antibody (Ab). We employed a 4.125-kb fragment of a mouse nephrin promoter downstream to which a FLAG-tagged hHO-1 cDNA sequence was inserted and subsequently generated transgenic mice from the FVB/N parental strain. There was a 16-fold higher transgene expression in the kidney than nonspecific background (liver) while the transprotein immunolocalized in glomerular epithelial cells (GEC). There was no change in urinary protein excretion, indicating that GEC-targeted HO-1 expression had no effect on glomerular protein permeability. Urinary protein excretion in transgenic mice with anti-GBM Ab injury (days 3 and 6) was significantly lower compared with wild-type controls. There was no significant change in renal expression levels of profibrotic (TGF-β1) or anti-inflammatory (IL-10) cytokines in transgenic mice with anti-GBM Ab injury. These observations indicate that GEC-targeted HO-1 expression does not alter glomerular physiological integrity and reduces proteinuria in glomerular immune injury. PMID:19587144

78 FR 17931 - Information Collection; Open Government Citizen Engagement Ratings, Rankings, and Flagging

Federal Register 2010, 2011, 2012, 2013, 2014

2013-03-25

...] Information Collection; Open Government Citizen Engagement Ratings, Rankings, and Flagging AGENCY: Office of... regarding open government citizen engagement ratings, rankings, and flagging. DATES: Comments must be...- 0288, Open Government Citizen Engagement Ratings, Rankings, and Flagging, by any of the following...

Interferon regulatory factor 1 and a variant of heterogeneous nuclear ribonucleoprotein L coordinately silence the gene for adhesion protein CEACAM1.

PubMed

Dery, Kenneth J; Silver, Craig; Yang, Lu; Shively, John E

2018-06-15

The adhesion protein carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) is widely expressed in epithelial cells as a short cytoplasmic isoform (S-iso) and in leukocytes as a long cytoplasmic isoform (L-iso) and is frequently silenced in cancer by unknown mechanisms. Previously, we reported that interferon response factor 1 (IRF1) biases alternative splicing (AS) to include the variable exon 7 (E7) in CEACAM1, generating long cytoplasmic isoforms. We now show that IRF1 and a variant of heterogeneous nuclear ribonucleoprotein L (Lv1) coordinately silence the CEACAM1 gene. RNAi-mediated Lv1 depletion in IRF1-treated HeLa and melanoma cells induced significant CEACAM1 protein expression, reversed by ectopic Lv1 expression. The Lv1-mediated CEACAM1 repression resided in residues Gly 71 -Gly 89 and Ala 38 -Gly 89 in Lv1's N-terminal extension. ChIP analysis of IRF1- and FLAG-tagged Lv1-treated HeLa cells and global treatment with the global epigenetic modifiers 5-aza-2'-deoxycytidine and trichostatin A indicated that IRF1 and Lv1 together induce chromatin remodeling, restricting IRF1 access to the CEACAM1 promoter. In interferon γ-treated HeLa cells, the transcription factor SP1 did not associate with the CEACAM1 promoter, but binding by upstream transcription factor 1 (USF1), a known CEACAM1 regulator, was greatly enhanced. ChIP-sequencing revealed that Lv1 overexpression in IRF1-treated cells induces transcriptional silencing across many genes, including DCC ( d eleted in c olorectal c arcinoma), associated with CEACAM5 in colon cancer. Notably, IRF1, but not IRF3 and IRF7, affected CEACAM1 expression via translational repression. We conclude that IRF1 and Lv1 coordinately regulate CEACAM1 transcription, alternative splicing, and translation and may significantly contribute to CEACAM1 silencing in cancer. © 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

SOCS2 Binds to and Regulates EphA2 through Multiple Mechanisms.

PubMed

Pilling, Carissa; Cooper, Jonathan A

2017-09-07

Suppressors of cytokine signaling (SOCS) proteins inhibit signaling by serving as substrate receptors for the Cullin5-RING E3 ubiquitin ligase (CRL5) and through a variety of CRL5-independent mechanisms. CRL5, SOCS2 and SOCS6 are implicated in suppressing transformation of epithelial cells. We identified cell proteins that interact with SOCS2 and SOCS6 using two parallel proteomics techniques: BioID and Flag affinity purification mass spectrometry. The receptor tyrosine kinase ephrin type-A receptor 2 (EphA2) was identified as a SOCS2-interacting protein. SOCS2-EphA2 binding requires the SOCS2 SH2 domain and EphA2 activation loop autophosphorylation, which is stimulated by Ephrin A1 (EfnA1) or by phosphotyrosine phosphatase inhibition. Surprisingly, EfnA1-stimulated EphA2-SOCS2 binding is delayed until EphA2 has been internalized into endosomes. This suggests that SOCS2 binds to EphA2 in the context of endosomal membranes. We also found that SOCS2 overexpression decreases steady state levels of EphA2, consistent with increased EphA2 degradation. This effect is indirect: SOCS2 induces EfnA1 expression, and EfnA1 induces EphA2 down-regulation. Other RTKs have been reported to bind, and be regulated by, over-expressed SOCS proteins. Our data suggest that SOCS protein over-expression may regulate receptor tyrosine kinases through indirect and direct mechanisms.

Mechanisms of flag-football injuries reported to the HQ Air Force Safety Center a 10-year descriptive study, 1993-2002.

PubMed

Burnham, Bruce R; Copley, G Bruce; Shim, Matthew J; Kemp, Philip A; Jones, Bruce H

2010-01-01

Flag (touch or intramural) football is a popular sport among the U.S. Air Force (USAF) active duty population and causes a substantial number of lost-workday injuries. The purpose of this study is to describe the mechanisms of flag-football injuries to better identify effective countermeasures. The data were derived from safety reports obtained from the USAF Ground Safety Automated System. Flag-football injuries for the years 1993-2002 that resulted in at least one lost workday were included in the study conducted in 2003. Narrative data were systematically reviewed for 32,812 USAF mishap reports; these were then coded in order to categorize and summarize mechanisms associated with flag football and other sports and occupational injuries. Nine hundred and forty-four mishap reports involving active duty USAF members playing flag football met the criteria for inclusion into this study. Eight mechanisms of injury were identified. The eight mechanisms accounted for 90% of all flag-football injuries. One scenario (contact with another player) accounted for 42% of all flag-football injuries. The most common mechanisms of injury caused by playing flag football can be identified using the detailed information found in safety reports. These scenarios are essential to developing evidence-based countermeasures. Results for flag football suggest that interventions that prevent player contact injuries deserve further research and evaluation. The broader implications of this study are that military safety data can be used to identify potentially modifiable mechanisms of injury for specific activities such as flag football. Published by Elsevier Inc.

76 FR 39885 - Risk-Based Targeting of Foreign Flagged Mobile Offshore Drilling Units (MODUs)

Federal Register 2010, 2011, 2012, 2013, 2014

2011-07-07

... Foreign Flagged Mobile Offshore Drilling Units (MODUs) AGENCY: Coast Guard, DHS. ACTION: Notice of... 11-06, Risk-Based Targeting of Foreign Flagged Mobile Offshore Drilling Units (MODUs). This policy... applicable regulations, every foreign-flagged mobile offshore drilling unit (MODU) must undergo a Coast Guard...

78 FR 36311 - Flag Recognition Benefit for Fallen Federal Civilian Employees

Federal Register 2010, 2011, 2012, 2013, 2014

2013-06-17

... Recognition Benefit for Fallen Federal Civilian Employees; Submission for Review: Application for U.S. Flag... PERSONNEL MANAGEMENT 5 CFR Part 550 RIN 3206-AM58 Flag Recognition Benefit for Fallen Federal Civilian... United States flag recognition benefit for fallen Federal civilian employees, and describe the...

Dicer-like 3 produces transposable element-associated 24-nt siRNAs that control agricultural traits in rice

PubMed Central

Wei, Liya; Gu, Lianfeng; Song, Xianwei; Cui, Xiekui; Lu, Zhike; Zhou, Ming; Wang, Lulu; Hu, Fengyi; Zhai, Jixian; Meyers, Blake C.; Cao, Xiaofeng

2014-01-01

Transposable elements (TEs) and repetitive sequences make up over 35% of the rice (Oryza sativa) genome. The host regulates the activity of different TEs by different epigenetic mechanisms, including DNA methylation, histone H3K9 methylation, and histone H3K4 demethylation. TEs can also affect the expression of host genes. For example, miniature inverted repeat TEs (MITEs), dispersed high copy-number DNA TEs, can influence the expression of nearby genes. In plants, 24-nt small interfering RNAs (siRNAs) are mainly derived from repeats and TEs. However, the extent to which TEs, particularly MITEs associated with 24-nt siRNAs, affect gene expression remains elusive. Here, we show that the rice Dicer-like 3 homolog OsDCL3a is primarily responsible for 24-nt siRNA processing. Impairing OsDCL3a expression by RNA interference caused phenotypes affecting important agricultural traits; these phenotypes include dwarfism, larger flag leaf angle, and fewer secondary branches. We used small RNA deep sequencing to identify 535,054 24-nt siRNA clusters. Of these clusters, ∼82% were OsDCL3a-dependent and showed significant enrichment of MITEs. Reduction of OsDCL3a function reduced the 24-nt siRNAs predominantly from MITEs and elevated expression of nearby genes. OsDCL3a directly targets genes involved in gibberellin and brassinosteroid homeostasis; OsDCL3a deficiency may affect these genes, thus causing the phenotypes of dwarfism and enlarged flag leaf angle. Our work identifies OsDCL3a-dependent 24-nt siRNAs derived from MITEs as broadly functioning regulators for fine-tuning gene expression, which may reflect a conserved epigenetic mechanism in higher plants with genomes rich in dispersed repeats or TEs. PMID:24554078

10 CFR 1002.3 - Custody of official seal and distinguishing flags.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 10 Energy 4 2011-01-01 2011-01-01 false Custody of official seal and distinguishing flags. 1002.3 Section 1002.3 Energy DEPARTMENT OF ENERGY (GENERAL PROVISIONS) OFFICIAL SEAL AND DISTINGUISHING FLAG General § 1002.3 Custody of official seal and distinguishing flags. The Secretary or his designee shall...

10 CFR 1002.3 - Custody of official seal and distinguishing flags.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 10 Energy 4 2010-01-01 2010-01-01 false Custody of official seal and distinguishing flags. 1002.3 Section 1002.3 Energy DEPARTMENT OF ENERGY (GENERAL PROVISIONS) OFFICIAL SEAL AND DISTINGUISHING FLAG General § 1002.3 Custody of official seal and distinguishing flags. The Secretary or his designee shall...

Interactomic approach for evaluating nucleophosmin-binding proteins as biomarkers for Ewing's sarcoma.

PubMed

Haga, Ayako; Ogawara, Yoko; Kubota, Daisuke; Kitabayashi, Issay; Murakami, Yasufumi; Kondo, Tadashi

2013-06-01

Nucleophosmin (NPM) is a novel prognostic biomarker for Ewing's sarcoma. To evaluate the prognostic utility of NPM, we conducted an interactomic approach to characterize the NPM protein complex in Ewing's sarcoma cells. A gene suppression assay revealed that NPM promoted cell proliferation and the invasive properties of Ewing's sarcoma cells. FLAG-tag-based affinity purification coupled with liquid chromatography-tandem mass spectrometry identified 106 proteins in the NPM protein complex. The functional classification suggested that the NPM complex participates in critical biological events, including ribosome biogenesis, regulation of transcription and translation, and protein folding, that are mediated by these proteins. In addition to JAK1, a candidate prognostic biomarker for Ewing's sarcoma, the NPM complex, includes 11 proteins known as prognostic biomarkers for other malignancies. Meta-analysis of gene expression profiles of 32 patients with Ewing's sarcoma revealed that 6 of 106 were significantly and independently associated with survival period. These observations suggest a functional role as well as prognostic value of these NPM complex proteins in Ewing's sarcoma. Further, our study suggests the potential applications of interactomics in conjunction with meta-analysis for biomarker discovery. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

The rescue and evaluation of FLAG and HIS epitope-tagged Asia 1 type foot-and-mouth disease viruses.

PubMed

Yang, Bo; Yang, Fan; Zhang, Yan; Liu, Huanan; Jin, Ye; Cao, Weijun; Zhu, Zixiang; Zheng, Haixue; Yin, Hong

2016-02-02

The VP1 G-H loop of the foot-and-mouth disease virus (FMDV) contains the primary antigenic site, as well as an Arg-Gly-Asp (RGD) binding motif for the αv-integrin family of cell surface receptors. We anticipated that introducing a foreign epitope tag sequence downstream of the RGD motif would be tolerated by the viral capsid and would not destroy the antigenic site of FMDV. In this study, we have designed, generated, and characterized two recombinant FMDVs with a FLAG tag or histidine (HIS) inserted in the VP1 G-H loop downstream of the RGD motif +9 position. The tagged viruses were genetically stable and exhibited similar growth properties with their parental virus. What is more, the recombinant viruses rFMDV-FLAG and rFMDV-HIS showed neutralization sensitivity to FMDV type Asia1-specific mAbs, as well as to polyclonal antibodies. Additionally, the r1 values of the recombinant viruses were similar to that of the parental virus, indicating that the insertion of FLAG or HIS tag sequences downstream of the RGD motif +9 position do not eradicate the antigenic site of FMDV and do not affect its antigenicity. These results indicated that the G-H loop of Asia1 FMDV is able to effectively display the foreign epitopes, making this a potential approach for novel FMDV vaccines development. Copyright © 2015 Elsevier B.V. All rights reserved.

Evaluation and optimization of the extended information process unit (E-IPU) validation module integrating the sysmex flag systems and the recommendations of the French-speaking cellular hematology group (GFHC).

PubMed

Cornet, Edouard; Mullier, François; Despas, Noemie; Jacqmin, Hugues; Geara, Carole; Boubaya, Marouane; Chatelain, Bernard; Troussard, Xavier

2016-10-01

The French-Speaking Cellular Haematology Group (GFHC) recently published criteria for microscopic analysis of a blood smears when a hemogram is requested. In order to evaluate and improve these recommendations using an XN (Sysmex) analyzer, we assessed 31,836 samples categorized into two sub-groups of patients either receiving or not receiving care in the clinical hematology/oncology departments of two university hospitals. By combining the manufacturer's recommendations and the GFHC recommendations, 21.3% of samples had a positive review flag in phase 1 of our study (17,991 samples). In phase 2 (13,845 samples), increasing the immature granulocytes (IG) percentage from 5-10% as a review trigger threshold, and ignoring slides with isolated flags 'PLT HIGH' (thrombocytosis) or 'MCV LOW' (microcytosis) or 'Blast/Abn Lymph and Atypical Lymph' (blast cells/abnormal lymphocytes and atypical lymphocytes) (in the absence of abnormal cells on a previous blood smear within 72 h), enabled us to significantly reduce the number of slides reviewed from 21.3-15.0% (p < 0.0001), without loss of clinical value. This decrease occurred in both sub-groups (hematology 48.7-38.0%, non-hematology 18.3-11.7%, p < 0.0001). In conclusion, the application of the GFHC criteria adapted to XN analyzers has enabled us to optimize the hematology laboratory processes, and thus reduce the production costs and the turnaround time of hemogram results.

Comparative architecture of silks, fibrous proteins and their encoding genes in insects and spiders.

PubMed

Craig, Catherine L; Riekel, Christian

2002-12-01

The known silk fibroins and fibrous glues are thought to be encoded by members of the same gene family. All silk fibroins sequenced to date contain regions of long-range order (crystalline regions) and/or short-range order (non-crystalline regions). All of the sequenced fibroin silks (Flag or silk from flagelliform gland in spiders; Fhc or heavy chain fibroin silks produced by Lepidoptera larvae) are made up of hierarchically organized, repetitive arrays of amino acids. Fhc fibroin genes are characterized by a similar molecular genetic architecture of two exons and one intron, but the organization and size of these units differs. The Flag, Ser (sericin gene) and BR (Balbiani ring genes; both fibrous proteins) genes are made up of multiple exons and introns. Sequences coding for crystalline and non-crystalline protein domains are integrated in the repetitive regions of Fhc and MA exons, but not in the protein glues Ser1 and BR-1. Genetic 'hot-spots' promote recombination errors in Fhc, MA, and Flag. Codon bias, structural constraint, point mutations, and shortened coding arrays may be alternative means of stabilizing precursor mRNA transcripts. Differential regulation of gene expression and selective splicing of the mRNA transcript may allow rapid adaptation of silk functional properties to different physical environments.

75 FR 34309 - Flag Day and National Flag Week, 2010

Federal Register 2010, 2011, 2012, 2013, 2014

2010-06-16

... Nation to confront tyranny and oppression still flies today as an unequivocal emblem of freedom and... gatherings to private memorials, we gathered to salute our flag, and in doing so, renewed the eternal promise... recognize the American flag as a symbol of hope and inspiration to people at home and around the world--as a...

Expressing the sense of the House of Representatives that a recipient of the Congressional Medal of Honor should be permitted, at all times on the recipient's property, to properly display the Flag of the United States of America.

THOMAS, 111th Congress

Rep. McKeon, Howard P. "Buck" [R-CA-25

2009-12-08

House - 06/15/2010 Referred to the Subcommittee on the Constitution, Civil Rights, and Civil Liberties. (All Actions) Tracker: This bill has the status IntroducedHere are the steps for Status of Legislation:

Localization and regulation of the N terminal splice variant of PGC-1α in adult skeletal muscle fibers.

PubMed

Shen, Tiansheng; Liu, Yewei; Schneider, Martin F

2012-01-01

The transcriptional coactivator peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α) regulates expression of genes for metabolism and muscle fiber type. Recently, a novel splice variant of PGC-1α (NT-PGC-1α, amino acids 1-270) was cloned and found to be expressed in muscle. Here we use Flag-tagged NT-PGC-1α to examine the subcellular localization and regulation of NT-PGC-1α in skeletal muscle fibers. Flag-NT-PGC-1α is located predominantly in the myoplasm. Nuclear NT-PGC-1α can be increased by activation of protein kinase A. Activation of p38 MAPK by muscle activity or of AMPK had no effect on the subcellular distribution of NT-PGC-1α. Inhibition of CRM1-mediated export only caused relatively slow nuclear accumulation of NT-PGC-1α, indicating that nuclear export of NT-PGC-1α may be mediated by both CRM1-dependent and -independent pathways. Together these results suggest that the regulation of NT-PGC-1α in muscle fibers may be very different from that of the full-length PGC-1α, which is exclusively nuclear.

A functional cutin matrix is required for plant protection against water loss

PubMed Central

Ma, Jian Feng; Li, Chao; Yamaji, Naoki; Nevo, Eviatar

2011-01-01

The plant cuticle, a cutin matrix embedded with and covered by wax, seals the aerial organ's surface to protect the plant against uncontrolled water loss. The cutin matrix is essential for the cuticle to function as a barrier to water loss. Recently, we identified from wild barley a drought supersensitive mutant, eibi1, which is caused by a defective cutin matrix as the result of the loss of function of HvABCG31, an ABCG full transporter. Here, we report that eibi1 epidermal cells contain lipid-like droplets, which are supposed to consist of cutin monomers that have not been transported out of the cells. The eibi1 cuticle is fragile due to a defective cutin matrix. The rice ortholog of the EIBI1 gene has a similar pattern of expression, young shoot but not flag leaf blade, as the barley gene. The model of the function of Eibi1 is discussed. The HvABCG31 full transporter functions in the export of cutin components and contributed to land plant colonization, hence also to terrestrial life evolution. PMID:22019635

A functional cutin matrix is required for plant protection against water loss.

PubMed

Chen, Guoxiong; Komatsuda, Takao; Ma, Jian Feng; Li, Chao; Yamaji, Naoki; Nevo, Eviatar

2011-09-01

The plant cuticle, a cutin matrix embedded with and covered by wax, seals the aerial organ's surface to protect the plant against uncontrolled water loss. The cutin matrix is essential for the cuticle to function as a barrier to water loss. Recently, we identified from wild barley a drought supersensitive mutant, eibi1, which is caused by a defective cutin matrix as the result of the loss of function of HvABCG31, an ABCG full transporter. Here, we report that eibi1 epidermal cells contain lipid-like droplets, which are supposed to consist of cutin monomers that have not been transported out of the cells. The eibi1 cuticle is fragile due to a defective cutin matrix. The rice ortholog of the EIBI1 gene has a similar pattern of expression, young shoot but not flag leaf blade, as the barley gene. The model of the function of Eibi1 is discussed. The HvABCG31 full transporter functions in the export of cutin components and contributed to land plant colonization, hence also to terrestrial life evolution.

78 FR 67432 - Self-Regulatory Organizations; EDGA Exchange, Inc.; Notice of Filing and Immediate Effectiveness...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-11-12

...) Increase the fee for orders yielding Flag K, which routes to NASDAQ OMX PSX (``PSX'') using ROUC or ROUE... for orders yielding Flag K, which routes to PSX using ROUC or ROUE routing strategies; and (ii) decrease the fee for orders yielding Flag RW, which routes to CBSX and adds liquidity. Flag K In securities...

3 CFR 8391 - Proclamation 8391 of June 11, 2009. Flag Day and National Flag Week, 2009

Code of Federal Regulations, 2010 CFR

2010-01-01

... America A Proclamation In the midst of a war for our Nation's independence, on June 14, 1777, the Second... America's promise and guides us toward a brighter tomorrow. To commemorate the adoption of our flag, the... toward equality and justice for all. Our flag's journey has been long. It has seen our Nation through war...

3 CFR 8689 - Proclamation 8689 of June 10, 2011. Flag Day and National Flag Week, 2011

Code of Federal Regulations, 2012 CFR

2012-01-01

... America A Proclamation On June 14, 1777, the Second Constitutional Congress adopted a flag with thirteen... were set upon a blue field, in the words of the Congress’s resolution, “representing a new... American flag has been ever present. It has flown on our ships and military bases around the world as we...

Red flag screening for low back pain: nothing to see here, move along: a narrative review.

PubMed

Cook, Chad E; George, Steven Z; Reiman, Michael P

2018-04-01

Screening for red flags in individuals with low back pain (LBP) has been a historical hallmark of musculoskeletal management. Red flag screening is endorsed by most LBP clinical practice guidelines, despite a lack of support for their diagnostic capacity. We share four major reasons why red flag screening is not consistent with best practice in LBP management: (1) clinicians do not actually screen for red flags, they manage the findings; (2) red flag symptomology negates the utility of clinical findings; (3) the tests lack the negative likelihood ratio to serve as a screen; and (4) clinical practice guidelines do not include specific processes that aid decision-making. Based on these findings, we propose that clinicians consider: (1) the importance of watchful waiting; (2) the value-based care does not support clinical examination driven by red flag symptoms; and (3) the recognition that red flag symptoms may have a stronger relationship with prognosis than diagnosis. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

Freestanding Flag-Type Triboelectric Nanogenerator for Harvesting High-Altitude Wind Energy from Arbitrary Directions.

PubMed

Zhao, Zhenfu; Pu, Xiong; Du, Chunhua; Li, Linxuan; Jiang, Chunyan; Hu, Weiguo; Wang, Zhong Lin

2016-02-23

Wind energy at a high altitude is far more stable and stronger than that near the ground, but it is out of reach of the wind turbine. Herein, we develop an innovative freestanding woven triboelectric nanogenerator flag (WTENG-flag) that can harvest high-altitude wind energy from arbitrary directions. The wind-driven fluttering of the woven unit leads to the current generation by a coupled effect of contact electrification and electrostatic induction. Systematic study is conducted to optimize the structure/material parameters of the WTENG-flag to improve the power output. This 2D WTENG-flag can also be stacked in parallel connections in many layers for a linearly increased output. Finally, a self-powered high-altitude platform with temperature/humidity sensing/telecommunicating capability is demonstrated with the WTENG-flag as a power source. Due to the light weight, low cost, and easy scale-up, this WTENG-flag has great potential for applications in weather/environmental sensing/monitoring systems.

Method for compression of binary data

DOEpatents

Berlin, Gary J.

1996-01-01

The disclosed method for compression of a series of data bytes, based on LZSS-based compression methods, provides faster decompression of the stored data. The method involves the creation of a flag bit buffer in a random access memory device for temporary storage of flag bits generated during normal LZSS-based compression. The flag bit buffer stores the flag bits separately from their corresponding pointers and uncompressed data bytes until all input data has been read. Then, the flag bits are appended to the compressed output stream of data. Decompression can be performed much faster because bit manipulation is only required when reading the flag bits and not when reading uncompressed data bytes and pointers. Uncompressed data is read using byte length instructions and pointers are read using word instructions, thus reducing the time required for decompression.

Development and validation of a pre-hospital "Red Flag" alert for activation of intra-hospital haemorrhage control response in blunt trauma.

PubMed

Hamada, Sophie Rym; Rosa, Anne; Gauss, Tobias; Desclefs, Jean-Philippe; Raux, Mathieu; Harrois, Anatole; Follin, Arnaud; Cook, Fabrice; Boutonnet, Mathieu; Attias, Arie; Ausset, Sylvain; Boutonnet, Mathieu; Dhonneur, Gilles; Duranteau, Jacques; Langeron, Olivier; Paugam-Burtz, Catherine; Pirracchio, Romain; de St Maurice, Guillaume; Vigué, Bernard; Rouquette, Alexandra; Duranteau, Jacques

2018-05-05

Haemorrhagic shock is the leading cause of early preventable death in severe trauma. Delayed treatment is a recognized prognostic factor that can be prevented by efficient organization of care. This study aimed to develop and validate Red Flag, a binary alert identifying blunt trauma patients with high risk of severe haemorrhage (SH), to be used by the pre-hospital trauma team in order to trigger an adequate intra-hospital standardized haemorrhage control response: massive transfusion protocol and/or immediate haemostatic procedures. A multicentre retrospective study of prospectively collected data from a trauma registry (Traumabase®) was performed. SH was defined as: packed red blood cell (RBC) transfusion in the trauma room, or transfusion ≥ 4 RBC in the first 6 h, or lactate ≥ 5 mmol/L, or immediate haemostatic surgery, or interventional radiology and/or death of haemorrhagic shock. Pre-hospital characteristics were selected using a multiple logistic regression model in a derivation cohort to develop a Red Flag binary alert whose performances were confirmed in a validation cohort. Among the 3675 patients of the derivation cohort, 672 (18%) had SH. The final prediction model included five pre-hospital variables: Shock Index ≥ 1, mean arterial blood pressure ≤ 70 mmHg, point of care haemoglobin ≤ 13 g/dl, unstable pelvis and pre-hospital intubation. The Red Flag alert was triggered by the presence of any combination of at least two criteria. Its predictive performances were sensitivity 75% (72-79%), specificity 79% (77-80%) and area under the receiver operating characteristic curve 0.83 (0.81-0.84) in the derivation cohort, and were not significantly different in the independent validation cohort of 2999 patients. The Red Flag alert developed and validated in this study has high performance to accurately predict or exclude SH.

Commercial Sealift and U.S. National Security

DTIC Science & Technology

2010-03-01

and maintaining a U.S. flag merchant marine fleet in today’s globalized shipping environment, where lower cost foreign flag registries of convenience ...in today’s globalized shipping environment, where lower cost foreign flag registries of convenience dominate the industry and which policy tools are...shipping environment, where lower cost foreign flag registries of convenience dominate the industry and which policy tools are best suited to meet our

American Colleges Raise the Flag in Vietnam

ERIC Educational Resources Information Center

Overland, Martha Ann

2009-01-01

More than 30 years after the U.S. ambassador was airlifted from the embassy rooftop in Saigon with the flag tucked under his arm, a new American flag is going up in the city. This one won't be flying over the embassy. The Stars and Stripes, as well as the Texas state flag, are going up at the Saigon Institute of Technology, the only Vietnamese…

77 FR 56890 - Self-Regulatory Organizations; EDGX Exchange, Inc.; Notice of Filing and Immediate Effectiveness...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-09-14

... PI, where Flag PI removes liquidity from the EDGX book against the Midpoint Match. This charge would signal a rate change for Flag PI if the conditions for achieving the Mega Tier \\4\\ are not satisfied. The Exchange also proposes to amend the text of Footnote 1 to add Flags BB and PI to the list of removal flags...

Progress toward an enhanced vaccine: Eight marked attenuated viruses to porcine reproductive and respiratory disease virus.

PubMed

Spear, Allyn; Wang, Feng-Xue; Kappes, Matthew A; Das, Phani B; Faaberg, Kay S

2018-03-01

Recombinant viruses of strain Ingelvac® PRRS porcine reproductive and respiratory syndrome virus (PRRSV) modified live virus vaccine were produced with two individual small in-frame deletions in nonstructural protein 2 (nsp2; Δ23 and Δ87) and also the same deletions supplanted with foreign tags (Δ23-V5, Δ23-FLAG, Δ23-S, Δ87-V5, Δ87-FLAG, Δ87-S). The viruses, but one (Δ87-FLAG), were stable for 10 passages and showed minimal effects on in vitro growth. Northern hybridization showed that the Δ23-tagged probe detected intracellular viral genome RNA as well as shorter RNAs that may represent heteroclite species, while the Δ87-tagged probe detected predominantly only genome length RNAs. When the tagged viruses were used to probe nsp2 protein in infected cells, perinuclear localization similar to native nsp2 was seen. Dual infection of Δ23-S and Δ87-S viruses allowed some discrimination of individual tagged nsp2 protein, facilitating future research. The mutants could potentially also be used to differentiate infected from vaccinated animals. Published by Elsevier Inc.

Brain Human Monoclonal Autoantibody from Sydenham Chorea Targets Dopaminergic Neurons in Transgenic Mice and Signals Dopamine D2 Receptor: Implications in Human Disease1

PubMed Central

Cox, Carol J.; Sharma, Meenakshi; Leckman, James F.; Zuccolo, Jonathan; Zuccolo, Amir; Kovoor, Abraham; Swedo, Susan E.; Cunningham, Madeleine W.

2013-01-01

How autoantibodies target the brain and lead to disease in disorders such as Sydenham chorea (SC) is not known. SC is characterized by autoantibodies against the brain and is the main neurologic manifestation of streptococcal-induced rheumatic fever. Previously, our novel SC-derived mAb 24.3.1 was found to recognize streptococcal and brain antigens. To investigate in vivo targets of human mAb 24.3.1, VH/VL genes were expressed in B cells of transgenic (Tg) mice as functional chimeric human VH 24.3.1 - mouse constant region IgG1a autoantibody. Chimeric human-mouse IgG1a autoantibody co-localized with tyrosine hydroxylase in the basal ganglia within dopaminergic neurons in vivo in VH 24.3.1 Tg mice. Both human mAb 24.3.1 and IgG1a in Tg sera were found to react with human dopamine D2 receptor (D2R). Reactivity of chorea-derived mAb 24.3.1 or SC IgG with D2R was confirmed by 1) dose dependent inhibitory signaling of D2R as a potential consequence of targeting dopaminergic neurons, 2) reaction with surface-exposed FLAG epitope-tagged D2R, and 3) blocking of Ab reactivity by an extracellular D2R peptide. IgG from SC and a related subset of streptococcal associated behavioral disorders called pediatric autoimmune neuropsychiatric disorder associated with streptococci (PANDAS) with small choreiform movements reacted in ELISA with D2R. Reaction with FLAG-tagged D2R distinguished SC from PANDAS while sera from both SC and PANDAS induced inhibitory signaling of D2R on transfected cells comparable to dopamine. Here we define a mechanism by which the brain may be altered by antibody in movement and behavioral disorders. PMID:24184556

Method for compression of binary data

DOEpatents

Berlin, G.J.

1996-03-26

The disclosed method for compression of a series of data bytes, based on LZSS-based compression methods, provides faster decompression of the stored data. The method involves the creation of a flag bit buffer in a random access memory device for temporary storage of flag bits generated during normal LZSS-based compression. The flag bit buffer stores the flag bits separately from their corresponding pointers and uncompressed data bytes until all input data has been read. Then, the flag bits are appended to the compressed output stream of data. Decompression can be performed much faster because bit manipulation is only required when reading the flag bits and not when reading uncompressed data bytes and pointers. Uncompressed data is read using byte length instructions and pointers are read using word instructions, thus reducing the time required for decompression. 5 figs.

Two Procedures to Flag Radio Frequency Interference in the UV Plane

NASA Astrophysics Data System (ADS)

Sekhar, Srikrishna; Athreya, Ramana

2018-07-01

We present two algorithms to identify and flag radio frequency interference (RFI) in radio interferometric imaging data. The first algorithm utilizes the redundancy of visibilities inside a UV cell in the visibility plane to identify corrupted data, while varying the detection threshold in accordance with the observed reduction in noise with radial UV distance. In the second algorithm, we propose a scheme to detect faint RFI in the visibility time-channel (TC) plane of baselines. The efficacy of identifying RFI in the residual visibilities is reduced by the presence of ripples due to inaccurate subtraction of the strongest sources. This can be due to several reasons including primary beam asymmetries and other direction-dependent calibration errors. We eliminated these ripples by clipping the corresponding peaks in the associated Fourier plane. RFI was detected in the ripple-free TC plane but was flagged in the original visibilities. Application of these two algorithms to five different 150 MHz data sets from the GMRT resulted in a reduction in image noise of 20%–50% throughout the field along with a reduction in systematics and a corresponding increase in the number of detected sources. However, in comparing the mean flux densities before and after flagging RFI, we find a differential change with the fainter sources (25σ < S < 100 mJy) showing a change of ‑6% to +1% relative to the stronger sources (S > 100 mJy). We are unable to explain this effect, but it could be related to the CLEAN bias known for interferometers.

Vertical Feature Mask Feature Classification Flag Extraction

Atmospheric Science Data Center

2013-03-28

  Vertical Feature Mask Feature Classification Flag Extraction This routine demonstrates extraction of the ... in a CALIPSO Lidar Level 2 Vertical Feature Mask feature classification flag value. It is written in Interactive Data Language (IDL) ...

The Identification of Novel Protein-Protein Interactions in Liver that Affect Glucagon Receptor Activity

PubMed Central

Froese, Sean; Dai, Feihan F.; Robitaille, Mélanie; Bhattacharjee, Alpana; Huang, Xinyi; Jia, Weiping; Angers, Stéphane; Wheeler, Michael B.; Wei, Li

2015-01-01

Glucagon regulates glucose homeostasis by controlling glycogenolysis and gluconeogenesis in the liver. Exaggerated and dysregulated glucagon secretion can exacerbate hyperglycemia contributing to type 2 diabetes (T2D). Thus, it is important to understand how glucagon receptor (GCGR) activity and signaling is controlled in hepatocytes. To better understand this, we sought to identify proteins that interact with the GCGR to affect ligand-dependent receptor activation. A Flag-tagged human GCGR was recombinantly expressed in Chinese hamster ovary (CHO) cells, and GCGR complexes were isolated by affinity purification (AP). Complexes were then analyzed by mass spectrometry (MS), and protein-GCGR interactions were validated by co-immunoprecipitation (Co-IP) and Western blot. This was followed by studies in primary hepatocytes to assess the effects of each interactor on glucagon-dependent glucose production and intracellular cAMP accumulation, and then in immortalized CHO and liver cell lines to further examine cell signaling. Thirty-three unique interactors were identified from the AP-MS screening of GCGR expressing CHO cells in both glucagon liganded and unliganded states. These studies revealed a particularly robust interaction between GCGR and 5 proteins, further validated by Co-IP, Western blot and qPCR. Overexpression of selected interactors in mouse hepatocytes indicated that two interactors, LDLR and TMED2, significantly enhanced glucagon-stimulated glucose production, while YWHAB inhibited glucose production. This was mirrored with glucagon-stimulated cAMP production, with LDLR and TMED2 enhancing and YWHAB inhibiting cAMP accumulation. To further link these interactors to glucose production, key gluconeogenic genes were assessed. Both LDLR and TMED2 stimulated while YWHAB inhibited PEPCK and G6Pase gene expression. In the present study, we have probed the GCGR interactome and found three novel GCGR interactors that control glucagon-stimulated glucose production by modulating cAMP accumulation and genes that control gluconeogenesis. These interactors may be useful targets to control glucose homeostasis in T2D. PMID:26075596

Participation of the extracellular domain in (pro)renin receptor dimerization

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

Suzuki-Nakagawa, Chiharu; Nishimura, Misa; Tsukamoto, Tomoko

Highlights: • The (pro)renin receptor [(P)RR] is a regulator of the renin–angiotensin system. • The region responsible for (P)RR dimerization was investigated. • (P)RR extracellular domain constructs were retained intracellularly. • The extracellular domain of (P)RR is responsible for its dimerization. • Novel insight into the regulatory mechanism of soluble (P)RR secretion is provided. - Abstract: The (pro)renin receptor [(P)RR] induces the catalytic activation of prorenin, as well as the activation of the mitogen-activated protein kinase (MAPK) signaling pathway; as such, it plays an important regulatory role in the renin–angiotensin system. (P)RR is known to form a homodimer, but themore » region participating in its dimerization is unknown. Using glutathione S-transferase (GST) as a carrier protein and a GST pull-down assay, we investigated the interaction of several (P)RR constructs with full-length (FL) (P)RR in mammalian cells. GST fusion proteins with FL (P)RR (GST-FL), the C-terminal M8-9 fragment (GST-M8-9), the extracellular domain (ECD) of (P)RR (GST-ECD), and the (P)RR ECD with a deletion of 32 amino acids encoded by exon 4 (GST-ECDd4) were retained intracellularly, whereas GST alone was efficiently secreted into the culture medium when transiently expressed in COS-7 cells. Immunofluorescence microscopy showed prominent localization of GST-ECD to the endoplasmic reticulum. The GST pull-down analysis revealed that GST-FL, GST-ECD, and GST-ECDd4 bound FLAG-tagged FL (P)RR, whereas GST-M8-9 showed little or no binding when transiently co-expressed in HEK293T cells. Furthermore, pull-down analysis using His-tag affinity resin showed co-precipitation of soluble (P)RR with FL (P)RR from a stable CHO cell line expressing FL h(P)RR with a C-terminal decahistidine tag. These results indicate that the (P)RR ECD participates in dimerization.« less

Heritage or Hate? A Pedagogical Guide to the Confederate Flag in Post-Race America

ERIC Educational Resources Information Center

Lippard, Cameron D.

2017-01-01

The Confederate flag has been a hotly debated symbol of heritage or hate in the United States. In 2015, 54 per cent of Americans polled saw the flag as a symbol of 'Southern pride' whereas 34 per cent saw it as racist. However, 27 per cent of Whites compared to 69 per cent of Blacks saw the flag as racist. In this article, I suggest how…

Flag-based detection of weak gas signatures in long-wave infrared hyperspectral image sequences

NASA Astrophysics Data System (ADS)

Marrinan, Timothy; Beveridge, J. Ross; Draper, Bruce; Kirby, Michael; Peterson, Chris

2016-05-01

We present a flag manifold based method for detecting chemical plumes in long-wave infrared hyperspectral movies. The method encodes temporal and spatial information related to a hyperspectral pixel into a flag, or nested sequence of linear subspaces. The technique used to create the flags pushes information about the background clutter, ambient conditions, and potential chemical agents into the leading elements of the flags. Exploiting this temporal information allows for a detection algorithm that is sensitive to the presence of weak signals. This method is compared to existing techniques qualitatively on real data and quantitatively on synthetic data to show that the flag-based algorithm consistently performs better on data when the SINRdB is low, and beats the ACE and MF algorithms in probability of detection for low probabilities of false alarm even when the SINRdB is high.

Identification of pixels with stray light and cloud shadow contaminations in the satellite ocean color data processing.

PubMed

Jiang, Lide; Wang, Menghua

2013-09-20

A new flag/masking scheme has been developed for identifying stray light and cloud shadow pixels that significantly impact the quality of satellite-derived ocean color products. Various case studies have been carried out to evaluate the performance of the new cloud contamination flag/masking scheme on ocean color products derived from the Visible Infrared Imaging Radiometer Suite (VIIRS) onboard the Suomi National Polar-orbiting Partnership (SNPP). These include direct visual assessments, detailed quantitative case studies, objective statistic analyses, and global image examinations and comparisons. The National Oceanic and Atmospheric Administration (NOAA) Multisensor Level-1 to Level-2 (NOAA-MSL12) ocean color data processing system has been used in the study. The new stray light and cloud shadow identification method has been shown to outperform the current stray light flag in both valid data coverage and data quality of satellite-derived ocean color products. In addition, some cloud-related flags from the official VIIRS-SNPP data processing software, i.e., the Interface Data Processing System (IDPS), have been assessed. Although the data quality with the IDPS flags is comparable to that of the new flag implemented in the NOAA-MSL12 ocean color data processing system, the valid data coverage from the IDPS is significantly less than that from the NOAA-MSL12 using the new stray light and cloud shadow flag method. Thus, the IDPS flag/masking algorithms need to be refined and modified to reduce the pixel loss, e.g., the proposed new cloud contamination flag/masking can be implemented in IDPS VIIRS ocean color data processing.

Defense.gov Special Report: Travels with Hagel

Science.gov Websites

Afghanistan Flag of Pakistan Flag of Saudi Arabia Flag of Qatar December 2013 News Stories Hagel Concludes Six -day Troop, Partner Nation Visits Defense Secretary Chuck Hagel wrapped up a six-day trip to the Middle

N-Glycosylation of the alpha subunit does not influence trafficking or functional activity of the human organic solute transporter alpha/beta

PubMed Central

Soroka, Carol J; Xu, Shuhua; Mennone, Albert; Lam, Ping; Boyer, James L

2008-01-01

Background The organic solute transporter (OSTα-OSTβ) is a heteromeric transporter that is expressed on the basolateral membrane of epithelium in intestine, kidney, liver, testis and adrenal gland and facilitates efflux of bile acids and other steroid solutes. Both subunits are required for plasma membrane localization of the functional transporter but it is unclear how and where the subunits interact and whether glycosylation is required for functional activity. We sought to examine these questions for the human OSTα-OSTβ transporter using the human hepatoma cell line, HepG2, and COS7 cells transfected with constructs of human OSTα-FLAG and OSTβ-Myc. Results Tunicamycin treatment demonstrated that human OSTα is glycosylated. In COS7 cells Western blotting identified the unglycosylated form (~31 kD), the core precursor form (~35 kD), and the mature, complex glycoprotein (~40 kD). Immunofluorescence of both cells indicated that, in the presence of OSTβ, the alpha subunit could still be expressed on the plasma membrane after tunicamycin treatment. Furthermore, the functional uptake of 3H-estrone sulfate was unchanged in the absence of N-glycosylation. Co-immunoprecipitation indicates that the immature form of OSTα interact with OSTβ. However, immunoprecipitation of OSTβ using an anti-Myc antibody did not co-precipitate the mature, complex glycosylated form of OSTα, suggesting that the primary interaction occurs early in the biosynthetic pathway and may be transient. Conclusion In conclusion, human OSTα is a glycoprotein that requires interaction with OSTβ to reach the plasma membrane. However, glycosylation of OSTα is not necessary for interaction with the beta subunit or for membrane localization or function of the heteromeric transporter. PMID:18847488

Identification and characterization of novel IGFBP5 interacting protein: evidence IGFBP5-IP is a potential regulator of osteoblast cell proliferation

PubMed Central

Amaar, Yousef G.; Tapia, Blanca; Chen, Shin-Tai; Baylink, David J.; Mohan, Subburaman

2010-01-01

Insulin-like growth factor binding protein-5 (IGFBP5) is a multifunctional protein, which acts not only as a traditional binding protein, but also functions as a growth factor independent of IGFs to stimulate bone formation. It has been predicted that the intrinsic growth factor action of IGFBP5 involves binding of IGFBP5 to a putative receptor to induce downstream signaling pathways and/or nuclear translocation of IGFBP5 to influence transcription of genes involved in osteoblast cell proliferation/differentiation. Our study indentified proteins that bound to IGFBP5 using IGFBP5 as bait in a yeast two-hybrid screen of the U2 human osteosarcoma cell cDNA library. One of the clones that interacted strongly with the bait under high-stringency conditions corresponded to a novel IGFBP5 interacting protein (IGFBP5-IP) encoded by a gene that resides in mouse chromosome 10. The interaction between IGFBP5-IP and IGFBP5 is confirmed by in vitro coimmunoprecipitation studies that used pFlag and IGFBP5 polyclonal antibody, and cell lysates overexpressing both IGFBP5-IP and IGFBP5. Northern blot and RT-PCR analysis showed that the IGFBP-IP is expressed in both untransformed normal human osteoblasts and in osteosarcoma cell lines, which are known to produce IGFBP5. To determine the roles of IGFBP5-IP, we evaluated the effect of blocking the expression of IGFBP5-IP on osteoblast proliferation. We found that using a IGFBP5-IP-specific small interfering-hairpin plasmid resulted in a decrease in both basal and IGFBP5-induced osteoblast cell proliferation. On the basis of these findings, we predict that IGFBP5-IP may act as intracellular mediator of growth promoting actions of IGFBP5 and perhaps other osteoregulatory agents in bone cells. PMID:16269403

46 CFR 151.45-9 - Signals.

Code of Federal Regulations, 2010 CFR

2010-10-01

... bulk cargo shall display a red flag by day or a red light by night, which signal shall be so placed... display a red flag by day, placed so that it will be visible on all sides. This flag may be metallic. ...

Guidelines for the specification of blue safety flags in railroad operations

DOT National Transportation Integrated Search

2010-12-01

Blue flag protection in the railroad industry provides safety to workers from the inadvertent movement of equipment on which they : are working. Current Federal regulations provide minimum specifications for the devices that can be used as blue flags...

Improved outcome in pediatric relapsed acute myeloid leukemia: results of a randomized trial on liposomal daunorubicin by the International BFM Study Group.

PubMed

Kaspers, Gertjan J L; Zimmermann, Martin; Reinhardt, Dirk; Gibson, Brenda E S; Tamminga, Rienk Y J; Aleinikova, Olga; Armendariz, Hortensia; Dworzak, Michael; Ha, Shau-Yin; Hasle, Henrik; Hovi, Liisa; Maschan, Alexei; Bertrand, Yves; Leverger, Guy G; Razzouk, Bassem I; Rizzari, Carmelo; Smisek, Petr; Smith, Owen; Stark, Batia; Creutzig, Ursula

2013-02-10

In pediatric relapsed acute myeloid leukemia (AML), optimal reinduction therapy is unknown. Studies suggest that liposomal daunorubicin (DNX; DaunoXome; Galen, Craigavon, United Kingdom) is effective and less cardiotoxic, which is important in this setting. These considerations led to a randomized phase III study by the International Berlin-Frankfurt-Münster Study Group. Patients with relapsed or primary refractory non-French-American-British type M3 AML who were younger than 21 years of age were eligible. Patients were randomly assigned to fludarabine, cytarabine, and granulocyte colony-stimulating factor (FLAG) or to FLAG plus DNX in the first reinduction course. The primary end point was status of the bone marrow (BM) sampled shortly before the second course of chemotherapy (the day 28 BM). Data are presented according to intention-to-treat for all 394 randomly assigned patients (median follow-up, 4.0 years). The complete remission (CR) rate was 64%, and the 4-year probability of survival (pOS) was 38% (SE, 3%). The day 28 BM status (available in 359 patients) was good (≤ 20% leukemic blasts) in 80% of patients randomly assigned to FLAG/DNX and 70% for patients randomly assigned to FLAG (P = .04). Concerning secondary end points, the CR rate was 69% with FLAG/DNX and 59% with FLAG (P = .07), but overall survival was similar. However, core-binding factor (CBF) AML treated with FLAG/DNX resulted in pOS of 82% versus 58% with FLAG (P = .04). Grade 3 to 4 toxicity was essentially similar in both groups. DNX added to FLAG improves early treatment response in pediatric relapsed AML. Overall long-term survival was similar, but CBF-AML showed an improved survival with FLAG/DNX. International collaboration proved feasible and resulted in the best outcome for pediatric relapsed AML reported thus far.

10 CFR 1.55 - Establishment of official NRC flag.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 10 Energy 1 2010-01-01 2010-01-01 false Establishment of official NRC flag. 1.55 Section 1.55 Energy NUCLEAR REGULATORY COMMISSION STATEMENT OF ORGANIZATION AND GENERAL INFORMATION NRC Seal and Flag... dark blue field with a gold fringe. ...

3 CFR 8993 - Proclamation 8993 of June 7, 2013. Flag Day and National Flag Week, 2013

Code of Federal Regulations, 2014 CFR

2014-01-01

... States to display the flag during that week. NOW, THEREFORE, I, BARACK OBAMA, President of the United... of the Independence of the United States of America the two hundred and thirty-seventh.BARACK OBAMA ...

Fake news portrayals of stem cells and stem cell research.

PubMed

Marcon, Alessandro R; Murdoch, Blake; Caulfield, Timothy

2017-10-01

This study examines how stem cells and stem cell research are portrayed on websites deemed to be purveyors of distorted and dubious information. Content analysis was conducted on 224 articles from 2015 to 2016, compiled by searching with the keywords 'stem cell(s)' on a list of websites flagged for containing either 'fake' or 'junk science' news. Articles contained various exaggerated positive and negative claims about stem cells and stem cell science, health and science related conspiracy theories, and statements promoting fear and mistrust of conventional medicine. Findings demonstrate the existence of organized misinformation networks, which may lead the public away from accurate information and facilitate a polarization of public discourse.

Predictive value of the present-on-admission indicator for hospital-acquired venous thromboembolism.

PubMed

Khanna, Raman R; Kim, Sharon B; Jenkins, Ian; El-Kareh, Robert; Afsarmanesh, Nasim; Amin, Alpesh; Sand, Heather; Auerbach, Andrew; Chia, Catherine Y; Maynard, Gregory; Romano, Patrick S; White, Richard H

2015-04-01

Hospital-acquired venous thromboembolic (HA-VTE) events are an important, preventable cause of morbidity and death, but accurately identifying HA-VTE events requires labor-intensive chart review. Administrative diagnosis codes and their associated "present-on-admission" (POA) indicator might allow automated identification of HA-VTE events, but only if VTE codes are accurately flagged "not present-on-admission" (POA=N). New codes were introduced in 2009 to improve accuracy. We identified all medical patients with at least 1 VTE "other" discharge diagnosis code from 5 academic medical centers over a 24-month period. We then sampled, within each center, patients with VTE codes flagged POA=N or POA=U (insufficient documentation) and POA=Y or POA=W (timing clinically uncertain) and abstracted each chart to clarify VTE timing. All events that were not clearly POA were classified as HA-VTE. We then calculated predictive values of the POA=N/U flags for HA-VTE and the POA=Y/W flags for non-HA-VTE. Among 2070 cases with at least 1 "other" VTE code, we found 339 codes flagged POA=N/U and 1941 flagged POA=Y/W. Among 275 POA=N/U abstracted codes, 75.6% (95% CI, 70.1%-80.6%) were HA-VTE; among 291 POA=Y/W abstracted events, 73.5% (95% CI, 68.0%-78.5%) were non-HA-VTE. Extrapolating from this sample, we estimated that 59% of actual HA-VTE codes were incorrectly flagged POA=Y/W. POA indicator predictive values did not improve after new codes were introduced in 2009. The predictive value of VTE events flagged POA=N/U for HA-VTE was 75%. However, sole reliance on this flag may substantially underestimate the incidence of HA-VTE.

The Savannah River Site`s Groundwater Monitoring Program. Second quarter, 1991

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

Not Available

1992-01-10

The Environmental Protection Department/Environmental Monitoring Section (EPD/EMS) administers the Savannah River Site`s (SRS) Groundwater Monitoring Program. During second quarter 1991 EPD/EMS conducted extensive sampling of monitoring wells. EPD/EMS established two sets of flagging criteria in 1986 to assist in the management of sample results. The flagging criteria do not define contamination levels; instead, they aid personnel in sample scheduling, interpretation of data, and trend identification. Beginning in 1991, the flagging criteria are based on EPA drinking water standards and method detection limits. A detailed explanation of the current flagging criteria is presented in the Flagging Criteria section of this document.more » Analytical results from second quarter 1991 are listed in this report.« less

The Savannah River Site's Groundwater Monitoring Program

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

Not Available

1992-01-10

The Environmental Protection Department/Environmental Monitoring Section (EPD/EMS) administers the Savannah River Site's (SRS) Groundwater Monitoring Program. During second quarter 1991 EPD/EMS conducted extensive sampling of monitoring wells. EPD/EMS established two sets of flagging criteria in 1986 to assist in the management of sample results. The flagging criteria do not define contamination levels; instead, they aid personnel in sample scheduling, interpretation of data, and trend identification. Beginning in 1991, the flagging criteria are based on EPA drinking water standards and method detection limits. A detailed explanation of the current flagging criteria is presented in the Flagging Criteria section of this document.more » Analytical results from second quarter 1991 are listed in this report.« less

KSC-2011-1496

NASA Image and Video Library

2011-02-18

CAPE CANAVERAL, Fla. -- "The National 9/11 Flag" is folded in the Rocket Garden at the Kennedy Space Center Visitor Complex in Florida. The contributions of NASA, Kennedy Space Center and the state of Florida were stitched into the fabric of the American Flag, which was recovered near ground zero following the World Trade Center attacks on Sept. 11, 2001. The "New York Says Thank You Foundation" is taking the flag on a cross-country journey to be restored to its original 13-stripe design using pieces of fabric from American flags destined for retirement in all 50 states. Once the flag is restored, it will become a permanent collection of the National September 11 Memorial Museum being built at the World Trade Center site. Photo credit: NASA/Kim Shiflett

KSC-2011-1484

NASA Image and Video Library

2011-02-18

CAPE CANAVERAL, Fla. -- "The National 9/11 Flag" is on display in the Debus Conference Facility at the Kennedy Space Center Visitor Complex in Florida. The contributions of NASA, Kennedy Space Center and the state of Florida were stitched into the fabric of the American Flag, which was recovered near ground zero following the World Trade Center attacks on Sept. 11, 2001. The "New York Says Thank You Foundation" is taking the flag on a cross-country journey to be restored to its original 13-stripe design using pieces of fabric from American flags destined for retirement in all 50 states. Once the flag is restored, it will become a permanent collection of the National September 11 Memorial Museum being built at the World Trade Center site. Photo credit: NASA/Kim Shiflett

KSC-2011-1495

NASA Image and Video Library

2011-02-18

CAPE CANAVERAL, Fla. -- "The National 9/11 Flag" is raised in the Rocket Garden at the Kennedy Space Center Visitor Complex in Florida. The contributions of NASA, Kennedy Space Center and the state of Florida were stitched into the fabric of the American Flag, which was recovered near ground zero following the World Trade Center attacks on Sept. 11, 2001. The "New York Says Thank You Foundation" is taking the flag on a cross-country journey to be restored to its original 13-stripe design using pieces of fabric from American flags destined for retirement in all 50 states. Once the flag is restored, it will become a permanent collection of the National September 11 Memorial Museum being built at the World Trade Center site. Photo credit: NASA/Kim Shiflett

77 FR 20098 - Inventory of U.S.-Flag Launch Barges

Federal Register 2010, 2011, 2012, 2013, 2014

2012-04-03

... DEPARTMENT OF TRANSPORTATION Maritime Administration [Docket No. MARAD-2012 0034] Inventory of U.S.-Flag Launch Barges AGENCY: Maritime Administration, Department of Transportation. ACTION: Inventory of U.S.-Flag Launch Barges. SUMMARY: The Maritime Administration is updating its inventory of U.S...

75 FR 13645 - Inventory of U.S.-Flag Launch Barges

Federal Register 2010, 2011, 2012, 2013, 2014

2010-03-22

... DEPARTMENT OF TRANSPORTATION Maritime Administration [Docket No. MARAD-2010 0023] Inventory of U.S.-Flag Launch Barges AGENCY: Maritime Administration, Department of Transportation. ACTION: Inventory of U.S.-Flag Launch Barges. SUMMARY: The Maritime Administration is updating its inventory of U.S...

Visual reproduction subtest of the Wechsler Memory Scale-Revised: analysis of construct validity.

PubMed

Williams, M A; Rich, M A; Reed, L K; Jackson, W T; LaMarche, J A; Boll, T J

1998-11-01

This study assessed the construct validity of Visual Reproduction (VR) Cards A (Flags) and B (Boxes) from the original Wechsler Memory Scale (WMS) compared to Flags and Boxes from the revised edition of the WMS (WMS-R). Independent raters scored Flags and Boxes using both the original and revised scoring criteria and correlations were obtained with age, education, IQ, and four separate criterion memory measures. Results show that for Flags, there is a tendency for the revised scoring criteria to produce improved construct validity. For Boxes, however, there was a trend in the opposite direction, with the revised scoring criteria demonstrating worse construct validity. Factor analysis suggests that Flags are a more distinct measure of visual memory, whereas Boxes are more complex and significantly associated with conceptual reasoning abilities. Using the revised scoring criteria, Boxes were found to be more strongly related to IQ than Flags. This difference was not found using the original scoring criteria.

FLAG - APOLLO XI - ASTRONAUTS - MOON

NASA Image and Video Library

1969-07-14

S69-39333 (July 1969) --- This is a photographic illustration of how the flag of the United States will be implanted on the moon by the Apollo 11 astronauts. The flag is three by five feet, and is made of nylon. It will be erected on an eight-foot aluminum staff, and tubing along its top edge will unfurl it in the airless environment of the moon. The implanting of the flag is symbolic of the first time man has landed on another celestial body, and does not constitute a territorial claim by the United States. The photograph on the right shows the flag in a furled condition. Apollo 11 astronauts Neil A. Armstrong, commander; and Edwin E. Aldrin Jr., lunar module pilot, will implant the flag after their Lunar Module (LM) sets down on the moon. Astronaut Michael Collins, command module pilot, will remain with the Command and Service Modules (CSM) in lunar orbit while Armstrong and Aldrin explore the lunar surface.

Molecular Modeling of Estrogen Receptor alpha Mutated Breast Cancer to Guide New Therapeutic Strategies

DTIC Science & Technology

2017-10-01

cancer cell lines that contain heterozygous ER LBD mutations and their wildtype controls. We first started this project using the standard CRISPR /Cas9...5’ HOM 3’ HOM Neomycin P Flag STOP CRISPR -mediated homologous recombination *mutant or WT ER Single cell cloning Screening MCF-7 D538G clones 6...Zannel Blanchard Grad. student ZBLANCHARD 12 Responsible for the molecular biology, CRISPR /Cas9 work. None Spencer Arnesen Grad. student

Association of CAD, a multifunctional protein involved in pyrimidine synthesis, with mLST8, a component of the mTOR complexes

PubMed Central

2013-01-01

Background mTOR is a genetically conserved serine/threonine protein kinase, which controls cell growth, proliferation, and survival. A multifunctional protein CAD, catalyzing the initial three steps in de novo pyrimidine synthesis, is regulated by the phosphorylation reaction with different protein kinases, but the relationship with mTOR protein kinase has not been known. Results CAD was recovered as a binding protein with mLST8, a component of the mTOR complexes, from HEK293 cells transfected with the FLAG-mLST8 vector. Association of these two proteins was confirmed by the co-immuoprecipitaiton followed by immunoblot analysis of transfected myc-CAD and FLAG-mLST8 as well as that of the endogenous proteins in the cells. Analysis using mutant constructs suggested that CAD has more than one region for the binding with mLST8, and that mLST8 recognizes CAD and mTOR in distinct ways. The CAD enzymatic activity decreased in the cells depleted of amino acids and serum, in which the mTOR activity is suppressed. Conclusion The results obtained indicate that mLST8 bridges between CAD and mTOR, and plays a role in the signaling mechanism where CAD is regulated in the mTOR pathway through the association with mLST8. PMID:23594158

RFI flagging implications for short-duration transients

NASA Astrophysics Data System (ADS)

Cendes, Y.; Prasad, P.; Rowlinson, A.; Wijers, R. A. M. J.; Swinbank, J. D.; Law, C. J.; van der Horst, A. J.; Carbone, D.; Broderick, J. W.; Staley, T. D.; Stewart, A. J.; Huizinga, F.; Molenaar, G.; Alexov, A.; Bell, M. E.; Coenen, T.; Corbel, S.; Eislöffel, J.; Fender, R.; Grießmeier, J.-M.; Jonker, P.; Kramer, M.; Kuniyoshi, M.; Pietka, M.; Stappers, B.; Wise, M.; Zarka, P.

2018-04-01

With their wide fields of view and often relatively long coverage of any position in the sky in imaging survey mode, modern radio telescopes provide a data stream that is naturally suited to searching for rare transients. However, Radio Frequency Interference (RFI) can show up in the data stream in similar ways to such transients, and thus the normal pre-treatment of filtering RFI (flagging) may also remove astrophysical transients from the data stream before imaging. In this paper we investigate how standard flagging affects the detectability of such transients by examining the case of transient detection in an observing mode used for Low Frequency Array (LOFAR; van Haarlem et al., 2013) surveys. We quantify the fluence range of transients that would be detected, and the reduction of their SNR due to partial flagging. We find that transients with a duration close to the integration sampling time, as well as bright transients with durations on the order of tens of seconds, are completely flagged. For longer transients on the order of several tens of seconds to minutes, the flagging effects are not as severe, although part of the signal is lost. For these transients, we present a modified flagging strategy which mitigates the effect of flagging on transient signals. We also present a script which uses the differences between the two strategies, and known differences between transient RFI and astrophysical transients, to notify the observer when a potential transient is in the data stream.

Tyrosine residues 654 and 670 in {beta}-cat enin are crucial in regulation of Met-{beta}-catenin interactions

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

Zeng, Gang; Apte, Udayan; Micsenyi, Amanda

2006-11-01

{beta}-catenin, a key component of the canonical Wnt pathway, is also regulated by tyrosine phosphorylation that regulates its association to E-cadherin. Previously, we reported its association with the hepatocyte growth factor (HGF) receptor Met at the membrane. HGF induced Met-{beta}-catenin dissociation and nuclear translocation of {beta}-catenin, which was tyrosine-phosphorylation-dependent. Here, we further investigate the Met-{beta}-catenin interaction by selectively mutating several tyrosine residues, alone or in combination, in {beta}-catenin. The mutants were subcloned into FLAG-CMV vector and stably transfected into rat hepatoma cells, which were treated with HGF. All single or double-mutant-transfected cells continued to show HGF-induced nuclear translocation of FLAG-{beta}-cateninmore » except the mutations affecting 654 and 670 simultaneously (Y654/670F), which coincided with the lack of formation of {beta}-catenin-TCF complex and DNA synthesis, in response to the HGF treatment. In addition, the Y654/670F-transfected cells also showed no phosphorylation of {beta}-catenin or dissociation from Met in response to HGF. Thus, intact 654 and 670 tyrosine residues in {beta}-catenin are crucial in HGF-mediated {beta}-catenin translocation, activation and mitogenesis.« less

Nanocarrier-mediated foliar zinc fertilization influences expression of metal homeostasis related genes in flag leaves and enhances gluten content in durum wheat.

PubMed

Deshpande, Paresh; Dapkekar, Ashwin; Oak, Manoj; Paknikar, Kishore; Rajwade, Jyutika

2018-01-01

Wheat is the staple food for most of the world's population; however, it is a poor source of zinc. Foliar fertilization of zinc via zinc loaded chitosan nanocarriers (Zn-CNP) post-anthesis has proved to be a promising approach for grain zinc enhancement in durum wheat as evidenced in our earlier study. However, the molecular mechanism of uptake of zinc via Zn-CNP remains unclear. Foliar application of Zn-CNP was performed at post anthesis stages in two durum wheat cultivars (MACS 3125 and UC1114, containing the Gpc-B1 gene), and expression levels of several metal-related genes were analyzed during early senescence. Zn-CNP application indeed caused changes in gene expression as revealed by qPCR data on representative genes involved in metal homeostasis, phloem transporters, and leaf senescence. Furthermore, zinc-regulated transporters and iron (Fe)-regulated transporter-like protein (ZIP) family [ZIP1, ZIP7, ZIP15], CA (carbonic anhydrase), and DMAS (2'-deoxymugineic acid synthase) in flag leaves exhibited significant correlation with zinc content in the seeds. The analysis of grain endosperm proteins showed enhancement of gamma gliadins while other gluten subunits decreased. Gene expression within ZIP family members varied with the type of cultivar mostly attributed to the Gpc-B1, concentration of external zinc ions as well as the type of tissue analyzed. Correlation analysis revealed the involvement of the selected genes in zinc enhancement. At the molecular level, uptake of zinc via Zn-CNP nanocarrier was comparable to the uptake of zinc via common zinc fertilizers i.e. ZnSO4.

46 CFR 154.22 - Foreign flag vessel: Certificate of Compliance endorsement application.

Code of Federal Regulations, 2010 CFR

2010-10-01

... requesting an endorsement for the carriage of ethylene oxide, a classification society certification that the... Commanding Officer, Marine Safety Center the plans, calculations, and information under § 154.15(b). [CGD 77... foreign flag vessel, whose flag administration issues IMO Certificates, must submit to the Commanding...

14 CFR 121.467 - Flight attendant duty period limitations and rest requirements: Domestic, flag, and supplemental...

Code of Federal Regulations, 2010 CFR

2010-01-01

... and rest requirements: Domestic, flag, and supplemental operations. 121.467 Section 121.467..., Flag, and Supplemental Operations § 121.467 Flight attendant duty period limitations and rest... attendant's home station, is not considered part of a rest period. (13) Each certificate holder conducting...

36 CFR 504.9 - Placards, signs, banners and flags.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 36 Parks, Forests, and Public Property 3 2010-07-01 2010-07-01 false Placards, signs, banners and flags. 504.9 Section 504.9 Parks, Forests, and Public Property SMITHSONIAN INSTITUTION RULES AND REGULATIONS GOVERNING SMITHSONIAN INSTITUTION BUILDINGS AND GROUNDS § 504.9 Placards, signs, banners and flags...

48 CFR 47.403-3 - Disallowance of expenditures.

Code of Federal Regulations, 2010 CFR

2010-10-01

... CONTRACT MANAGEMENT TRANSPORTATION Air Transportation by U.S.-Flag Carriers 47.403-3 Disallowance of... air transportation on foreign-flag air carriers unless there is attached to the appropriate voucher a memorandum adequately explaining why service by U.S.-flag air carriers was not available, or why it was...

49 CFR 218.37 - Flag protection.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 49 Transportation 4 2010-10-01 2010-10-01 false Flag protection. 218.37 Section 218.37..., DEPARTMENT OF TRANSPORTATION RAILROAD OPERATING PRACTICES Protection of Trains and Locomotives § 218.37 Flag protection. (a) After August 1, 1977, each railroad must have in effect an operating rule which complies with...

36 CFR 520.10 - Placards, signs, banners, and flags.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 36 Parks, Forests, and Public Property 3 2010-07-01 2010-07-01 false Placards, signs, banners, and flags. 520.10 Section 520.10 Parks, Forests, and Public Property SMITHSONIAN INSTITUTION RULES AND... § 520.10 Placards, signs, banners, and flags. The displaying or carrying of placards, signs, banners, or...

VAB Flag Painting

NASA Image and Video Library

2002-01-01

On platforms suspended from the top of the 525-foot-high VAB, workers use rollers and brushes to repaint the U.S. flag on the southwest side of the Vehicle Assembly Building. The flag spans an area 209 feet by 110 feet, or about 23, 437 square feet. Each stripe is 9 feet wide and each star is 6 feet in diameter. The logo is also being painted. Known as the "meatball," the logo measures 110 feet by 132 feet, or about 12,300 square feet. The flag and logo were last painted in 1998, honoring NASA's 40th anniversary.

KSC-2011-1493

NASA Image and Video Library

2011-02-18

CAPE CANAVERAL, Fla. -- "The National 9/11 Flag" is transported from the Debus Conference Facility to the Rocket Garden at the Kennedy Space Center Visitor Complex in Florida. The contributions of NASA, Kennedy Space Center and the state of Florida were stitched into the fabric of the American Flag, which was recovered near ground zero following the World Trade Center attacks on Sept. 11, 2001. The "New York Says Thank You Foundation" is taking the flag on a cross-country journey to be restored to its original 13-stripe design using pieces of fabric from American flags destined for retirement in all 50 states. Once the flag is restored, it will become a permanent collection of the National September 11 Memorial Museum being built at the World Trade Center site. Photo credit: NASA/Kim Shiflett

KSC-2011-1494

NASA Image and Video Library

2011-02-18

CAPE CANAVERAL, Fla. -- "The National 9/11 Flag" is transported from the Debus Conference Facility to the Rocket Garden at the Kennedy Space Center Visitor Complex in Florida. The contributions of NASA, Kennedy Space Center and the state of Florida were stitched into the fabric of the American Flag, which was recovered near ground zero following the World Trade Center attacks on Sept. 11, 2001. The "New York Says Thank You Foundation" is taking the flag on a cross-country journey to be restored to its original 13-stripe design using pieces of fabric from American flags destined for retirement in all 50 states. Once the flag is restored, it will become a permanent collection of the National September 11 Memorial Museum being built at the World Trade Center site. Photo credit: NASA/Kim Shiflett

KSC-2011-1497

NASA Image and Video Library

2011-02-18

CAPE CANAVERAL, Fla. -- More than a dozen 9/11 first responders take part in "The National 9/11 Flag" stitching ceremony at the Kennedy Space Center Visitor Complex in Florida. The contributions of NASA, Kennedy Space Center and the state of Florida were stitched into the fabric of the American Flag, which was recovered near ground zero following the World Trade Center attacks on Sept. 11, 2001. The "New York Says Thank You Foundation" is taking the flag on a cross-country journey to be restored to its original 13-stripe design using pieces of fabric from American flags destined for retirement in all 50 states. Once the flag is restored, it will become a permanent collection of the National September 11 Memorial Museum being built at the World Trade Center site. Photo credit: NASA/Kim Shiflett

Electron microscopy and in vitro deneddylation reveal similar architectures and biochemistry of isolated human and Flag-mouse COP9 signalosome complexes

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

Rockel, Beate; Schmaler, Tilo; Huang, Xiaohua

2014-07-25

Highlights: • Deneddylation rates of human erythrocyte and mouse fibroblast CSN are very similar. • 3D models of native human and mouse CSN reveal common architectures. • The cryo-structure of native mammalian CSN shows a horseshoe subunit arrangement. - Abstract: The COP9 signalosome (CSN) is a regulator of the ubiquitin (Ub) proteasome system (UPS). In the UPS, proteins are Ub-labeled for degradation by Ub ligases conferring substrate specificity. The CSN controls a large family of Ub ligases called cullin-RING ligases (CRLs), which ubiquitinate cell cycle regulators, transcription factors and DNA damage response proteins. The CSN possesses structural similarities with themore » 26S proteasome Lid complex and the translation initiation complex 3 (eIF3) indicating similar ancestry and function. Initial structures were obtained 14 years ago by 2D electron microscopy (EM). Recently, first 3D molecular models of the CSN were created on the basis of negative-stain EM and single-particle analysis, mostly with recombinant complexes. Here, we compare deneddylating activity and structural features of CSN complexes purified in an elaborate procedure from human erythrocytes and efficiently pulled down from mouse Flag-CSN2 B8 fibroblasts. In an in vitro deneddylation assay both the human and the mouse CSN complexes deneddylated Nedd8-Cul1 with comparable rates. 3D structural models of the erythrocyte CSN as well as of the mouse Flag-CSN were generated by negative stain EM and by cryo-EM. Both complexes show a central U-shaped segment from which several arms emanate. This structure, called the horseshoe, is formed by the PCI domain subunits. CSN5 and CSN6 point away from the horseshoe. Compared to 3D models of negatively stained CSN complexes, densities assigned to CSN2 and CSN4 are better defined in the cryo-map. Because biochemical and structural results obtained with CSN complexes isolated from human erythrocytes and purified by Flag-CSN pulldown from mouse B8 fibroblasts are very similar, Flag-CSN pulldowns are a proper alternative to CSN preparation from erythrocytes.« less

Parallel Exploration of Interaction Space by BioID and Affinity Purification Coupled to Mass Spectrometry.

PubMed

Hesketh, Geoffrey G; Youn, Ji-Young; Samavarchi-Tehrani, Payman; Raught, Brian; Gingras, Anne-Claude

2017-01-01

Complete understanding of cellular function requires knowledge of the composition and dynamics of protein interaction networks, the importance of which spans all molecular cell biology fields. Mass spectrometry-based proteomics approaches are instrumental in this process, with affinity purification coupled to mass spectrometry (AP-MS) now widely used for defining interaction landscapes. Traditional AP-MS methods are well suited to providing information regarding the temporal aspects of soluble protein-protein interactions, but the requirement to maintain protein-protein interactions during cell lysis and AP means that both weak-affinity interactions and spatial information is lost. A more recently developed method called BioID employs the expression of bait proteins fused to a nonspecific biotin ligase, BirA*, that induces in vivo biotinylation of proximal proteins. Coupling this method to biotin affinity enrichment and mass spectrometry negates many of the solubility and interaction strength issues inherent in traditional AP-MS methods, and provides unparalleled spatial context for protein interactions. Here we describe the parallel implementation of both BioID and FLAG AP-MS allowing simultaneous exploration of both spatial and temporal aspects of protein interaction networks.

Intra-domain phage display (ID-PhD) of peptides and protein mini-domains censored from canonical pIII phage display.

PubMed

Tjhung, Katrina F; Deiss, Frédérique; Tran, Jessica; Chou, Ying; Derda, Ratmir

2015-01-01

In this paper, we describe multivalent display of peptide and protein sequences typically censored from traditional N-terminal display on protein pIII of filamentous bacteriophage M13. Using site-directed mutagenesis of commercially available M13KE phage cloning vector, we introduced sites that permit efficient cloning using restriction enzymes between domains N1 and N2 of the pIII protein. As infectivity of phage is directly linked to the integrity of the connection between N1 and N2 domains, intra-domain phage display (ID-PhD) allows for simple quality control of the display and the natural variations in the displayed sequences. Additionally, direct linkage to phage propagation allows efficient monitoring of sequence cleavage, providing a convenient system for selection and evolution of protease-susceptible or protease-resistant sequences. As an example of the benefits of such an ID-PhD system, we displayed a negatively charged FLAG sequence, which is known to be post-translationally excised from pIII when displayed on the N-terminus, as well as positively charged sequences which suppress production of phage when displayed on the N-terminus. ID-PhD of FLAG exhibited sub-nanomolar apparent Kd suggesting multivalent nature of the display. A TEV-protease recognition sequence (TEVrs) co-expressed in tandem with FLAG, allowed us to demonstrate that 99.9997% of the phage displayed the FLAG-TEVrs tandem and can be recognized and cleaved by TEV-protease. The residual 0.0003% consisted of phage clones that have excised the insert from their genome. ID-PhD is also amenable to display of protein mini-domains, such as the 33-residue minimized Z-domain of protein A. We show that it is thus possible to use ID-PhD for multivalent display and selection of mini-domain proteins (Affibodies, scFv, etc.).

The effects of a novel aliphatic-chain hydroxamate derivative WMJ-S-001 in HCT116 colorectal cancer cell death

PubMed Central

Huang, Yu-Han; Huang, Shiu-Wen; Hsu, Ya-Fen; Ou, George; Huang, Wei-Jan; Hsu, Ming-Jen

2015-01-01

Hydroxamate derivatives have attracted considerable attention due to their broad pharmacological properties and have been extensively investigated. We recently demonstrated that WMJ-S-001, a novel aliphatic hydroxamate derivative, exhibits anti-inflammatory and anti-angiogenic activities. In this study, we explored the underlying mechanisms by which WMJ-S-001 induces HCT116 colorectal cancer cell death. WMJ-S-001 inhibited cell proliferation and induced cell apoptosis in HCT116 cells. These actions were associated with AMP-activated protein kinase (AMPK) and p38 mitogen-activated protein kinase (MAPK) activation, p53 phosphorylation and acetylation, as well as the modulation of p21cip/Waf1, cyclin D1, survivin and Bax. AMPK-p38MAPK signaling blockade reduced WMJ-S-001-induced p53 phosphorylation. Transfection with AMPK dominant negative mutant (DN) reduced WMJ-S-001’s effects on p53 and Sp1 binding to the survivn promoter region. Transfection with HDAC3-Flag or HDAC4-Flag also abrogated WMJ-S-001’s enhancing effect on p53 acetylation. WMJ-S-001’s actions on p21cip/Waf1, cyclin D1, survivin, Bax were reduced in p53-null HCT116 cells. Furthermore, WMJ-S-001 was shown to suppress the growth of subcutaneous xenografts of HCT116 cells in vivo. In summary, the death of HCT116 colorectal cancer cells exposed to WMJ-S-001 may involve AMPK-p38MAPK-p53-survivin cascade. These results support the role of WMJ-S-001 as a potential drug candidate and warrant the clinical development in the treatment of cancer. PMID:26510776

Genetic dissection and validation of candidate genes for flag leaf size in rice (Oryza sativa L.).

PubMed

Tang, Xinxin; Gong, Rong; Sun, Wenqiang; Zhang, Chaopu; Yu, Sibin

2018-04-01

Two major loci with functional candidate genes were identified and validated affecting flag leaf size, which offer desirable genes to improve leaf architecture and photosynthetic capacity in rice. Leaf size is a major determinant of plant architecture and yield potential in crops. However, the genetic and molecular mechanisms regulating leaf size remain largely elusive. In this study, quantitative trait loci (QTLs) for flag leaf length and flag leaf width in rice were detected with high-density single nucleotide polymorphism genotyping of a chromosomal segment substitution line (CSSL) population, in which each line carries one or a few chromosomal segments from the japonica cultivar Nipponbare in a common background of the indica variety Zhenshan 97. In total, 14 QTLs for flag leaf length and nine QTLs for flag leaf width were identified in the CSSL population. Among them, qFW4-2 for flag leaf width was mapped to a 37-kb interval, with the most likely candidate gene being the previously characterized NAL1. Another major QTL for both flag leaf width and length was delimited by substitution mapping to a small region of 13.5 kb that contains a single gene, Ghd7.1. Mutants of Ghd7.1 generated using CRISPR/CAS9 approach showed reduced leaf size. Allelic variation analyses also validated Ghd7.1 as a functional candidate gene for leaf size, photosynthetic capacity and other yield-related traits. These results provide useful genetic information for the improvement of leaf size and yield in rice breeding programs.

Critical components of the pluripotency network are targets for the p300/CBP interacting protein (p/CIP) in embryonic stem cells.

PubMed

Chitilian, J M; Thillainadesan, G; Manias, J L; Chang, W Y; Walker, E; Isovic, M; Stanford, W L; Torchia, J

2014-01-01

p/CIP, also known as steroid receptor coactivator 3 (SRC-3)/Nuclear Receptor Coactivator 3 (NCoA3), is a transcriptional coactivator that binds liganded nuclear hormone receptors, as well as other transcription factors, and facilitates transcription through direct recruitment of accessory factors. We have found that p/CIP is highly expressed in undifferentiated mouse embryonic stem cells (mESCs) and is downregulated during differentiation. siRNA-mediated knockdown of p/CIP decreased transcript levels of Nanog, but not Oct4 or Sox2. Microarray expression analysis showed that Klf4, Tbx3, and Dax-1 are significantly downregulated in mESCs when p/CIP is knocked down. Subsequent chromatin immunoprecipitation (ChIP) analysis demonstrated that Tbx3, Klf4, and Dax-1 are direct transcriptional targets of p/CIP. Using the piggyBac transposition system, a mouse ESC line that expresses Flag-p/CIP in a doxycycline-dependent manner was generated. p/CIP overexpression increased the level of target genes and promoted the formation of undifferentiated colonies. Collectively, these results indicate that p/CIP contributes to the maintenance of ESC pluripotency through direct regulation of essential pluripotency genes. To better understand the mechanism by which p/CIP functions in ESC pluripotency, we integrated our ChIP and transcriptome data with published protein-protein interaction and promoter occupancy data to draft a p/CIP gene regulatory network. The p/CIP gene regulatory network identifies various feed-forward modules including one in which p/CIP activates members of the extended pluripotency network, demonstrating that p/CIP is a component of this extended network. © AlphaMed Press.

14 CFR 121.601 - Aircraft dispatcher information to pilot in command: Domestic and flag operations.

Code of Federal Regulations, 2011 CFR

2011-01-01

... command: Domestic and flag operations. 121.601 Section 121.601 Aeronautics and Space FEDERAL AVIATION... Flight Release Rules § 121.601 Aircraft dispatcher information to pilot in command: Domestic and flag operations. (a) The aircraft dispatcher shall provide the pilot in command all available current reports or...

14 CFR 121.601 - Aircraft dispatcher information to pilot in command: Domestic and flag operations.

Code of Federal Regulations, 2013 CFR

2013-01-01

... command: Domestic and flag operations. 121.601 Section 121.601 Aeronautics and Space FEDERAL AVIATION... Flight Release Rules § 121.601 Aircraft dispatcher information to pilot in command: Domestic and flag operations. (a) The aircraft dispatcher shall provide the pilot in command all available current reports or...

14 CFR 121.601 - Aircraft dispatcher information to pilot in command: Domestic and flag operations.

Code of Federal Regulations, 2012 CFR

2012-01-01

... command: Domestic and flag operations. 121.601 Section 121.601 Aeronautics and Space FEDERAL AVIATION... Flight Release Rules § 121.601 Aircraft dispatcher information to pilot in command: Domestic and flag operations. (a) The aircraft dispatcher shall provide the pilot in command all available current reports or...

14 CFR 121.601 - Aircraft dispatcher information to pilot in command: Domestic and flag operations.

Code of Federal Regulations, 2014 CFR

2014-01-01

... command: Domestic and flag operations. 121.601 Section 121.601 Aeronautics and Space FEDERAL AVIATION... Flight Release Rules § 121.601 Aircraft dispatcher information to pilot in command: Domestic and flag operations. (a) The aircraft dispatcher shall provide the pilot in command all available current reports or...

14 CFR 121.601 - Aircraft dispatcher information to pilot in command: Domestic and flag operations.

Code of Federal Regulations, 2010 CFR

2010-01-01

... command: Domestic and flag operations. 121.601 Section 121.601 Aeronautics and Space FEDERAL AVIATION... Flight Release Rules § 121.601 Aircraft dispatcher information to pilot in command: Domestic and flag operations. (a) The aircraft dispatcher shall provide the pilot in command all available current reports or...

14 CFR 121.641 - Fuel supply: Nonturbine and turbo-propeller-powered airplanes: Flag operations.

Code of Federal Regulations, 2014 CFR

2014-01-01

...-powered airplanes: Flag operations. 121.641 Section 121.641 Aeronautics and Space FEDERAL AVIATION... Flight Release Rules § 121.641 Fuel supply: Nonturbine and turbo-propeller-powered airplanes: Flag operations. (a) No person may dispatch or take off a nonturbine or turbo-propeller-powered airplane unless...

14 CFR 121.641 - Fuel supply: Nonturbine and turbo-propeller-powered airplanes: Flag operations.

Code of Federal Regulations, 2012 CFR

2012-01-01

...-powered airplanes: Flag operations. 121.641 Section 121.641 Aeronautics and Space FEDERAL AVIATION... Flight Release Rules § 121.641 Fuel supply: Nonturbine and turbo-propeller-powered airplanes: Flag operations. (a) No person may dispatch or take off a nonturbine or turbo-propeller-powered airplane unless...

14 CFR 121.641 - Fuel supply: Nonturbine and turbo-propeller-powered airplanes: Flag operations.

Code of Federal Regulations, 2013 CFR

2013-01-01

...-powered airplanes: Flag operations. 121.641 Section 121.641 Aeronautics and Space FEDERAL AVIATION... Flight Release Rules § 121.641 Fuel supply: Nonturbine and turbo-propeller-powered airplanes: Flag operations. (a) No person may dispatch or take off a nonturbine or turbo-propeller-powered airplane unless...

14 CFR 121.641 - Fuel supply: Nonturbine and turbo-propeller-powered airplanes: Flag operations.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 14 Aeronautics and Space 3 2010-01-01 2010-01-01 false Fuel supply: Nonturbine and turbo-propeller-powered airplanes: Flag operations. 121.641 Section 121.641 Aeronautics and Space FEDERAL AVIATION... Flight Release Rules § 121.641 Fuel supply: Nonturbine and turbo-propeller-powered airplanes: Flag...

14 CFR 121.641 - Fuel supply: Nonturbine and turbo-propeller-powered airplanes: Flag operations.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 14 Aeronautics and Space 3 2011-01-01 2011-01-01 false Fuel supply: Nonturbine and turbo-propeller-powered airplanes: Flag operations. 121.641 Section 121.641 Aeronautics and Space FEDERAL AVIATION... Flight Release Rules § 121.641 Fuel supply: Nonturbine and turbo-propeller-powered airplanes: Flag...

77 FR 35807 - Flag Day and National Flag Week, 2012

Federal Register 2010, 2011, 2012, 2013, 2014

2012-06-15

.... Generations of service members have raised our country's colors over military bases and at sea, and... Day and National Flag Week, 2012 By the President of the United States of America A Proclamation... Woodrow Wilson asked us to ``stand with united hearts for an America which no man can corrupt, no...

76 FR 35087 - Flag Day and National Flag Week, 2011

Federal Register 2010, 2011, 2012, 2013, 2014

2011-06-15

... United States of America A Proclamation On June 14, 1777, the Second Constitutional Congress adopted a... founding colonies. The stars were set upon a blue field, in the words of the Congress's resolution... faced, the American flag has been ever present. It has flown on our ships and military bases around the...

14 CFR 121.621 - Alternate airport for destination: Flag operations.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 14 Aeronautics and Space 3 2013-01-01 2013-01-01 false Alternate airport for destination: Flag... § 121.621 Alternate airport for destination: Flag operations. (a) No person may dispatch an airplane under IFR or over-the-top unless he lists at least one alternate airport for each destination airport in...

14 CFR 121.621 - Alternate airport for destination: Flag operations.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 14 Aeronautics and Space 3 2011-01-01 2011-01-01 false Alternate airport for destination: Flag... § 121.621 Alternate airport for destination: Flag operations. (a) No person may dispatch an airplane under IFR or over-the-top unless he lists at least one alternate airport for each destination airport in...

14 CFR 121.621 - Alternate airport for destination: Flag operations.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 14 Aeronautics and Space 3 2012-01-01 2012-01-01 false Alternate airport for destination: Flag... § 121.621 Alternate airport for destination: Flag operations. (a) No person may dispatch an airplane under IFR or over-the-top unless he lists at least one alternate airport for each destination airport in...

14 CFR 121.621 - Alternate airport for destination: Flag operations.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 14 Aeronautics and Space 3 2010-01-01 2010-01-01 false Alternate airport for destination: Flag... § 121.621 Alternate airport for destination: Flag operations. (a) No person may dispatch an airplane under IFR or over-the-top unless he lists at least one alternate airport for each destination airport in...

14 CFR 121.621 - Alternate airport for destination: Flag operations.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 14 Aeronautics and Space 3 2014-01-01 2014-01-01 false Alternate airport for destination: Flag... § 121.621 Alternate airport for destination: Flag operations. (a) No person may dispatch an airplane under IFR or over-the-top unless he lists at least one alternate airport for each destination airport in...

Fraction Flags: Learning from Children to Help Children Learn.

ERIC Educational Resources Information Center

Kieren, Tom; And Others

1996-01-01

Describes "fraction flags", an activity through which fraction concepts can be explored. The activity was invented by 2 12-year-old students and this article is presented with emphasis on the students' viewpoint. It begins with an overview of the fractions unit and presents vignettes of students exploring the fraction flags. (AIM)

KSC-2011-1485

NASA Image and Video Library

2011-02-18

CAPE CANAVERAL, Fla. -- Members of the Brevard Police and Fire Pipes and Drums kick off the "The National 9/11 Flag" stitching ceremony in the Debus Conference Facility at the Kennedy Space Center Visitor Complex in Florida. The contributions of NASA, Kennedy Space Center and the state of Florida were stitched into the fabric of the American Flag, which was recovered near ground zero following the World Trade Center attacks on Sept. 11, 2001. The "New York Says Thank You Foundation" is taking the flag on a cross-country journey to be restored to its original 13-stripe design using pieces of fabric from American flags destined for retirement in all 50 states. Once the flag is restored, it will become a permanent collection of the National September 11 Memorial Museum being built at the World Trade Center site. Photo credit: NASA/Kim Shiflett

KSC-2011-1489

NASA Image and Video Library

2011-02-18

CAPE CANAVERAL, Fla. -- Chief of Fire Training George Hoggard with NASA Kennedy Space Center Protective Services contributes stitches to the "National 9/11 Flag" during a ceremony in the Debus Conference Facility at the Kennedy Space Center Visitor Complex in Florida. The contributions of NASA, Kennedy Space Center and the state of Florida were stitched into the fabric of the American Flag, which was recovered near ground zero following the World Trade Center attacks on Sept. 11, 2001. The "New York Says Thank You Foundation" is taking the flag on a cross-country journey to be restored to its original 13-stripe design using pieces of fabric from American flags destined for retirement in all 50 states. Once the flag is restored, it will become a permanent collection of the National September 11 Memorial Museum being built at the World Trade Center site. Photo credit: NASA/Kim Shiflett

KSC-2011-1491

NASA Image and Video Library

2011-02-18

CAPE CANAVERAL, Fla. -- Kelvin Manning, associate director for Business Operations at NASA's Kennedy Space Center, contributes stitches to the "National 9/11 Flag" during a ceremony in the Debus Conference Facility at the Kennedy Space Center Visitor Complex in Florida. The contributions of NASA, Kennedy Space Center and the state of Florida were stitched into the fabric of the American Flag, which was recovered near ground zero following the World Trade Center attacks on Sept. 11, 2001. The "New York Says Thank You Foundation" is taking the flag on a cross-country journey to be restored to its original 13-stripe design using pieces of fabric from American flags destined for retirement in all 50 states. Once the flag is restored, it will become a permanent collection of the National September 11 Memorial Museum being built at the World Trade Center site. Photo credit: NASA/Kim Shiflett

KSC-2011-1490

NASA Image and Video Library

2011-02-18

CAPE CANAVERAL, Fla. -- Joe Dowdy, special operations manager at NASA's Kennedy Space Center, contributes stitches to the "National 9/11 Flag" during a ceremony in the Debus Conference Facility at the Kennedy Space Center Visitor Complex in Florida. The contributions of NASA, Kennedy Space Center and the state of Florida were stitched into the fabric of the American Flag, which was recovered near ground zero following the World Trade Center attacks on Sept. 11, 2001. The "New York Says Thank You Foundation" is taking the flag on a cross-country journey to be restored to its original 13-stripe design using pieces of fabric from American flags destined for retirement in all 50 states. Once the flag is restored, it will become a permanent collection of the National September 11 Memorial Museum being built at the World Trade Center site. Photo credit: NASA/Kim Shiflett

KSC-2011-1486

NASA Image and Video Library

2011-02-18

CAPE CANAVERAL, Fla. -- Members of the Brevard Police and Fire Pipes and Drums kick off the "The National 9/11 Flag" stitching ceremony in the Debus Conference Facility at the Kennedy Space Center Visitor Complex in Florida. The contributions of NASA, Kennedy Space Center and the state of Florida were stitched into the fabric of the American Flag, which was recovered near ground zero following the World Trade Center attacks on Sept. 11, 2001. The "New York Says Thank You Foundation" is taking the flag on a cross-country journey to be restored to its original 13-stripe design using pieces of fabric from American flags destined for retirement in all 50 states. Once the flag is restored, it will become a permanent collection of the National September 11 Memorial Museum being built at the World Trade Center site. Photo credit: NASA/Kim Shiflett

Utilising flags to reduce drag around a short finite circular cylinder

NASA Astrophysics Data System (ADS)

Javadi, Kh.; Kiani, F.; Tahaye Abadi, M.

2018-03-01

This paper utilises flags to decrease the drag around a short finite circular cylinder. Wall-adapted large eddy simulation and two-way fluid-structure interaction methods were applied to resolve unsteady turbulent flow structure. The far-field Reynolds number of the current configuration based on the cylinder diameter was chosen to be 20,000. In addition, the length-to-diameter ratio of the cylinder was assumed to be L/D = 2 whereas the flexible flag had a width-to-diameter ratio of W/D = 1.5. The results were compared with the regular short finite circular cylinder and the rigid flagged cylinder in our previous work. The results indicate that utilising flags inside the near-wake region of the cylinder reduces the pressure drag. The physical mechanism of this drag reduction is presented.

Red flag findings in children with headaches: Prevalence and association with emergency department neuroimaging.

PubMed

Tsze, Daniel S; Ochs, Julie B; Gonzalez, Ariana E; Dayan, Peter S

2018-01-01

Background Clinicians appear to obtain emergent neuroimaging for children with headaches based on the presence of red flag findings. However, little data exists regarding the prevalence of these findings in emergency department populations, and whether the identification of red flag findings is associated with potentially unnecessary emergency department neuroimaging. Objectives We aimed to determine the prevalence of red flag findings and their association with neuroimaging in otherwise healthy children presenting with headaches to the emergency department. Our secondary aim was to determine the prevalence of emergent intracranial abnormalities in this population. Methods A prospective cohort study of otherwise healthy children 2-17 years of age presenting to an urban pediatric emergency department with non-traumatic headaches was undertaken. Emergency department physicians completed a standardized form to document headache descriptors and characteristics, associated symptoms, and physical and neurological exam findings. Children who did not receive emergency department neuroimaging received 4-month telephone follow-up. Outcomes included emergency department neuroimaging and the presence of emergent intracranial abnormalities. Results We enrolled 224 patients; 197 (87.9%) had at least one red flag finding on history. Several red flag findings were reported by more than a third of children, including: Headache waking from sleep (34.8%); headache present with or soon after waking (39.7%); or headaches increasing in frequency, duration and severity (40%, 33.1%, and 46.3%). Thirty-three percent of children received emergency department neuroimaging. The prevalence of emergent intracranial abnormalities was 1% (95% CI 0.1, 3.6). Abnormal neurological exam, extreme pain intensity of presenting headache, vomiting, and positional symptoms were independently associated with emergency department neuroimaging. Conclusions Red flag findings are common in children presenting with headaches to the emergency department. The presence of red flag findings is associated with emergency department neuroimaging, although the risk of emergent intracranial abnormalities is low. Many children with headaches may be receiving unnecessary neuroimaging due to the high prevalence of non-specific red flag findings.

VAB Flag Painting

NASA Image and Video Library

2002-01-01

On platforms suspended from the top of the 525-foot-high VAB, workers use rollers and brushes to repaint the NASA logo on the southeast side of the Vehicle Assembly Building. Known as the "meatball," the logo measures 110 feet by 132 feet, or about 12,300 square feet. The U.S. flag is also being repainted. The flag spans an area 209 feet by 110 feet, or about 23, 437 square feet. Each stripe is 9 feet wide and each star is 6 feet in diameter. The flag and logo were last painted in 1998, honoring NASA's 40th anniversary.

New quantitative trait loci in wheat for flag leaf resistance to Stagonospora nodorum blotch.

PubMed

Francki, M G; Shankar, M; Walker, E; Loughman, R; Golzar, H; Ohm, H

2011-11-01

Stagonospora nodorum blotch (SNB) is a significant disease in some wheat-growing regions of the world. Resistance in wheat to Stagonospora nodorum is complex, whereby genes for seedling, flag leaf, and glume resistance are independent. The aims of this study were to identify alternative genes for flag leaf resistance, to compare and contrast with known quantitative trait loci (QTL) for SNB resistance, and to determine the potential role of host-specific toxins for SNB QTL. Novel QTL for flag leaf resistance were identified on chromosome 2AS inherited from winter wheat parent 'P92201D5' and chromosome 1BS from spring wheat parent 'EGA Blanco'. The chromosomal map position of markers associated with QTL on 1BS and 2AS indicated that they were unlikely to be associated with known host-toxin insensitivity loci. A QTL on chromosome 5BL inherited from EGA Blanco had highly significant association with markers fcp001 and fcp620 based on disease evaluation in 2007 and, therefore, is likely to be associated with Tsn1-ToxA insensitivity for flag leaf resistance. However, fcp001 and fcp620 were not associated with a QTL detected based on disease evaluation in 2008, indicating two linked QTL for flag leaf resistance with multiple genes residing on 5BL. This study identified novel QTL and their effects in controlling flag leaf SNB resistance.

Generation and characterization of a human-mouse chimeric high-affinity antibody that detects the DYKDDDDK FLAG peptide.

PubMed

Ikeda, Koki; Koga, Tomoaki; Sasaki, Fumiyuki; Ueno, Ayumi; Saeki, Kazuko; Okuno, Toshiaki; Yokomizo, Takehiko

2017-05-13

DYKDDDDK peptide (FLAG) is a useful tool for investigating the function and localization of proteins whose antibodies (Abs) are not available. We recently established a high-affinity monoclonal antibody (mAb) for FLAG (clone 2H8). The 2H8 Ab is highly sensitive for detecting FLAG-tagged proteins by flowcytometry and immunoprecipitation, but it can yield nonspecific signals in immunohistochemistry of mouse tissues because it is of mouse origin. In this study, we reduced nonspecific signals by generating a chimeric 2H8 Ab with Fc fragments derived from human immunoglobulin. We fused a 5' terminal cDNA fragments for the Fab region of 2H8 mAb with 3' terminal cDNA fragments for Fc region of human IgG1. We transfected both chimeric plasmids and purified the resulting human-mouse chimeric 2H8. The chimeric 2H8 Ab successfully detected FLAG-tagged proteins in flowcytometry with anti-human IgG secondary Ab with comparable sensitivity to 2H8 mAb. Importantly, chimeric 2H8 detected specific FLAG peptide signals without nonspecific signals in immunohistochemical analysis with mouse tissues. This human-mouse chimeric high-affinity anti-FLAG Ab will prove useful for future immunohistochemical analysis of mouse tissues. Copyright © 2017 Elsevier Inc. All rights reserved.

OsNucleolin1-L Expression in Arabidopsis Enhances Photosynthesis via Transcriptome Modification under Salt Stress Conditions.

PubMed

Udomchalothorn, Thanikarn; Plaimas, Kitiporn; Sripinyowanich, Siriporn; Boonchai, Chutamas; Kojonna, Thammaporn; Chutimanukul, Panita; Comai, Luca; Buaboocha, Teerapong; Chadchawan, Supachitra

2017-04-01

OsNUC1 encodes rice nucleolin, which has been shown to be involved in salt stress responses. Expression of the full-length OsNUC1 gene in Arabidopsis resulted in hypersensitivity to ABA during germination. Transcriptome analysis of the transgenic lines, in comparison with the wild type, revealed that the RNA abundance of >1,900 genes was significantly changed under normal growth conditions, while under salt stress conditions the RNAs of 999 genes were found to be significantly regulated. Gene enrichment analysis showed that under normal conditions OsNUC1 resulted in repression of genes involved in photosynthesis, while in salt stress conditions OsNUC1 increased expression of the genes involved in the light-harvesting complex. Correspondingly, the net rate of photosynthesis of the transgenic lines was increased under salt stress. Transgenic rice lines with overexpression of the OsNUC1-L gene were generated and tested for photosynthetic performance under salt stress conditions. The transgenic rice lines treated with salt stress at the booting stage had a higher photosynthetic rate and stomatal conductance in flag leaves and second leaves than the wild type. Moreover, higher contents of Chl a and carotenoids were found in flag leaves of the transgenic rice. These results suggest a role for OsNUC1 in the modification of the transcriptome, especially the gene transcripts responsible for photosynthesis, leading to stabilization of photosynthesis under salt stress conditions. © The Author 2017. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

Defense.gov - Special Report: Travels with Panetta

Science.gov Websites

Flag of Japan Japan Flag of China China Flag of New Zealand New Zealand Top Stories Secretary Honors Past, Present New Zealand Troops Defense Secretary Leon E. Panetta placed a wreath in memory of New Memorial Museum. Story Panetta Eases Restrictions on New Zealand Ship Visits Defense Secretary Leon E

3 CFR 8535 - Proclamation 8535 of June 11, 2010. Flag Day and National Flag Week, 2010

Code of Federal Regulations, 2011 CFR

2011-01-01

..., the thirteen stripes alternating red and white, and thirteen white stars in a blue field, represented... luminosity, and the enduring American story that it represents. Although the configuration of stars and... first embraced by our Founders, the Stars and Stripes remain the symbol of our Nation’s pride. On Flag...

FlagHouse Forum: You Say "Tomato"... and I Use a Communicator

ERIC Educational Resources Information Center

Exceptional Parent, 2011

2011-01-01

This month's "FlagHouse Forum" focuses on how to choose the communicator best-suited to a child's special need. FlagHouse--a premier global supplier of resources for special needs, education, physical activity and recreation--is pleased to partner with "Exceptional Parent" to bring its readers this informational forum. Humans communicate with each…

22 CFR 201.15 - U.S. flag vessel shipping requirements.

Code of Federal Regulations, 2011 CFR

2011-04-01

... and tankers shall be achieved for each quantitative unit of cargo. A quantitative unit of cargo is the... determined that at least 50% of the quantitative unit will move on U.S. flag vessels, to the extent that such... used for achieving compliance for the quantitative unit. (c) Nonavailability of U.S. flag vessels. Upon...

22 CFR 201.15 - U.S. flag vessel shipping requirements.

Code of Federal Regulations, 2014 CFR

2014-04-01

... and tankers shall be achieved for each quantitative unit of cargo. A quantitative unit of cargo is the... determined that at least 50% of the quantitative unit will move on U.S. flag vessels, to the extent that such... used for achieving compliance for the quantitative unit. (c) Nonavailability of U.S. flag vessels. Upon...

22 CFR 201.15 - U.S. flag vessel shipping requirements.

Code of Federal Regulations, 2010 CFR

2010-04-01

... and tankers shall be achieved for each quantitative unit of cargo. A quantitative unit of cargo is the... determined that at least 50% of the quantitative unit will move on U.S. flag vessels, to the extent that such... used for achieving compliance for the quantitative unit. (c) Nonavailability of U.S. flag vessels. Upon...

22 CFR 201.15 - U.S. flag vessel shipping requirements.

Code of Federal Regulations, 2012 CFR

2012-04-01

... and tankers shall be achieved for each quantitative unit of cargo. A quantitative unit of cargo is the... determined that at least 50% of the quantitative unit will move on U.S. flag vessels, to the extent that such... used for achieving compliance for the quantitative unit. (c) Nonavailability of U.S. flag vessels. Upon...

22 CFR 201.15 - U.S. flag vessel shipping requirements.

Code of Federal Regulations, 2013 CFR

2013-04-01

... and tankers shall be achieved for each quantitative unit of cargo. A quantitative unit of cargo is the... determined that at least 50% of the quantitative unit will move on U.S. flag vessels, to the extent that such... used for achieving compliance for the quantitative unit. (c) Nonavailability of U.S. flag vessels. Upon...

78 FR 67430 - Self-Regulatory Organizations; EDGX Exchange, Inc.; Notice of Filing and Immediate Effectiveness...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-11-12

... yield Flag K would increase intermarket competition because it offers customers an alternative means to...) Increase the fee for orders yielding Flag K, which routes to NASDAQ OMX PSX (``PSX'') using ROUC or ROUE... for orders yielding Flag K, which routes to PSX using ROUC or ROUE routing strategies; and (ii...

FIRE! A Red Flag Tap in Reclaiming Intervention

ERIC Educational Resources Information Center

Bodnar, Brian

2007-01-01

"Red Flag Interventions" address problems which are imported from elsewhere and acted out towards persons who are in effect innocent bystanders. This is commonly seen as students "carry in" problems from the home or street to school, or they "carry over" conflicts from one class to the next. A third variation of Red Flag intervention is when a…

78 FR 23617 - Self-Regulatory Organizations; EDGX Exchange, Inc.; Notice of Filing and Immediate Effectiveness...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-04-19

... Exchange introduced new Flags ZA (Retail Order, adds liquidity) and ZR (Retail Order, removes liquidity... will enable Members, and in turn, their retail customers, to benefit from the enhanced rebate (Flag ZA... able to benefit from the rebate (Flag ZA) for utilizing Retail Orders without regards to whether the...

Red flags: a case series of clinician-family communication challenges in the context of CHD.

PubMed

Sekar, Priya; Marcus, Katie L; Williams, Erin P; Boss, Renee D

2017-07-01

We describe three cases of newborns with complex CHD characterised by communication challenges. These communication challenges were categorised as patient, family, or system-related red flags. Strategies for addressing these red flags were proposed, for the goal of optimising care and improving quality of life in this vulnerable population.

75 FR 61836 - Additional Designation of Individuals and Entities Pursuant to Executive Order 13382

Federal Register 2010, 2011, 2012, 2013, 2014

2010-10-06

.... ABTIN 1 Container Ship 13,760DWT 9,957GRT IRAN flag (IRISL); Vessel Registration Identification IMO... IMO 9405954 (Malta) (vessel) [NPWMD]. 8. EIGHTH OCEAN General Cargo 22,882DWT 15,670GRT GERMANY flag... Container Ship 85,896DWT 74,175GRT MALTA flag (IRISL); Vessel Registration Identification IMO 9349576 (Malta...

3 CFR 8837 - Proclamation 8837 of June 11, 2012. Flag Day and National Flag Week, 2012

Code of Federal Regulations, 2013 CFR

2013-01-01

... homes and storefronts. Generations of service members have raised our country's colors over military... America A Proclamation Ninety-six years ago, our Nation first came together to celebrate Flag Day—an occasion when President Woodrow Wilson asked us to “stand with united hearts for an America which no man...

VLA Hosts "Flag Across America"

NASA Astrophysics Data System (ADS)

2001-11-01

The National Radio Astronomy Observatory (NRAO) hosted the runners and support personnel of the "Americans United Flag Across America" run as the transcontinental memorial and fundraising effort came through New Mexico. The flag run arrived at NRAO's Very Large Array (VLA) radio telescope west of Socorro, NM, early in the post-Midnight morning of Monday, November 5, and departed after sunrise that morning en route to the Arizona border. Drivers, runners and support personnel stayed overnight at the VLA. During the night, a "VLA Night Owl Run" kept the flag moving around the VLA area until the westward trek resumed after dawn. The run began Oct. 11, one month after the terrorist attacks on New York and Washington. Organized by employees of American and United Airlines to honor the flight crews lost in those attacks, to show support for U.S. troops and to raise funds to help the victims' families, the run will take an American flag from Boston Logan Airport to Los Angeles International Airport. The Boston-to-Los Angeles trip represents the intended journey of American Flight 11 and United Flight 175, both of which were crashed by terrorists into the World Trade Center. "Our observatory was proud to host this group and honored that they brought this flag through our facility," said Miller Goss, NRAO's director of VLA operations. The runners carried a flag that flew in a U.S. F-16 over Iraq in support of Operation Southern Watch on Oct. 2, and has visited Ground Zero in Manhattan. The flag is scheduled to arrive in Los Angeles on Veterans Day, Nov. 11. The National Radio Astronomy Observatory is a facility of the National Science Foundation, operated under cooperative agreement by Associated Universities, Inc.

Association between community socioeconomic characteristics and access to youth flag football.

PubMed

Kroshus, Emily; Sonnen, Aly J; Chrisman, Sara Pd; Rivara, Frederick P

2018-01-12

The American Academy of Pediatrics has recommended that opportunities for non-tackling American football (e.g., flag football) be expanded, given concerns about the risks of brain trauma from tackle football. This study tested the hypothesis that flag football would be more accessible in communities characterised by higher socioeconomic status residents. In July 2017, the locations of community-based organisations offering youth flag and tackle football for youth between the ages of 6 and 13 in two US states (Georgia and Washington) were aggregated (n=440). Organisations were coded in terms of the availability of tackle and/or flag football teams for youth at each year of age between 6 and 13. Multivariate logistic regression analyses were used to assess the odds of a community-based football organisation offering flag football, by community socioeconomic and demographic characteristics. In both states, communities with more educated residents were more likely to offer flag football for youth aged 6-12. For example, among 6 year-olds every 10% increase in the number of adult residents with a college education was associated with 1.51 times the odds of flag football availability (95% CI 1.22 to 1.86, P<0.001). These results suggest that youth living in communities characterised by low educational attainment are less likely than other youth to have the option of a lower contact alternative to tackle football. Relying on voluntary community-level adoption of lower contact alternatives to tackle football may result in inequitable access to such sport options. This may contribute to an inequitable burden of brain trauma from youth sport. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

Biochemical and mass spectrometric characterization of the human CB2 cannabinoid receptor expressed in Pichia pastoris--importance of correct processing of the N-terminus.

PubMed

Zhang, Rundong; Kim, Tae-Kang; Qiao, Zhaun-Hong; Cai, Jian; Pierce, William M; Song, Zhao-Hui

2007-10-01

This study was conducted to optimize the expression of human CB2 cannabinoid receptors in methylotrophic yeast Pichia pastoris (P. pastoris). Two major species of expressed CB2 proteins were seen on Western blot, i.e., a 42 kDa band which matches the calculated molecular weight for tagged CB2, and a 52/55 kDa doublet. Treatment of membranes with N-glycosidase F or inclusion of tunicamycin in the culture medium during induction resulted in the disappearance of the 55 kDa, but not the 52 kDa band, suggesting that the 3 kDa extra in the 55 kDa band is due to N-glycosylation, but the 10 kDa extra in the 52 kDa band is not due to N-glycosylation. Anti-FLAG M1 antibody had a much higher preference for the 42 kDa band over the 52/55 kDa doublet, and a 10 kDa fragment recognized by anti-FLAG M2 antibody was generated by CNBr digestion of the 52/55 doublet. These data strongly support the hypothesis that the 10 kDa increase in molecular weight was due to unprocessed alpha-factor sequence. This conclusion was further validated by finding several peptide sequences for alpha-factor fragments at the N-terminal of the CB2 receptor using pepsin/chymotrypsin digestion and LC/MS/MS approaches. Importantly, unprocessed alpha-factor was found to be associated with poor ligand binding. In addition, controlling the level of CB2 protein expression was found to be critical for minimizing the presence of unprocessed alpha-factor sequence. The information gained from this study should aid the proper expression of not only CB2 receptor but also other members of the GPCR family in P. pastoris.

Lithobates catesbeianus (American Bullfrog) oocytes: a novel heterologous expression system for aquaporins

PubMed Central

2018-01-01

ABSTRACT Xenopus laevis oocytes are a valuable tool for investigating the function of membrane proteins. However, regulations around the world, specifically in Brazil, render the import of Xenopus laevis frogs impractical, and, in some cases, impossible. Here, as an alternative, we evaluate the usefulness of the North American aquatic bullfrog Lithobates catesebeianus, which is commercially available in Brazil, for the heterologous expression of aquaporin (AQP) proteins. We have developed a method that combines a brief collagenase treatment and mechanical defolliculation for isolating individual oocytes from Lithobates ovaries. We find that they have a similar size, shape, and appearance to Xenopus oocytes and can tolerate and survive following injections with cRNA or water. Furthermore, surface biotinylation, western blot analysis, and measurements of osmotic water permeability (Pf) show that Lithobates oocytes can express AQPs to the plasma membrane and significantly increase the Pf of the oocytes. In fact, the Pf values are similar to historical values gathered from Xenopus oocytes. Due to the presence of a mercury sensitive cysteine (Cys or C) in the throat of the water channel, the Pf of oocytes expressing human (h) AQP1, hAQP1FLAG [FLAG, short protein tag (DYKDDDDK) added to the N-terminus of AQP1], hAQP8, and rat (r) AQP9 was inhibited with the mercurial compound p-chloromercuribenzene sulfonate (pCMBS), whereas AQPs lacking this Cys – hAQP1C189S mutant [residue Cys 189 was replaced by a serine (Ser or S)] and hAQP7 – were mercury insensitive. Contrary to previous studies with Xenopus oocytes, rAQP3 was also found to be insensitive to mercury, which is consistent with the mercury-sensitive Cys (Cys 11) being located intracellularly. Thus, we consider Lithobates oocytes to be a readily accessible system for the functional expression and study of membrane proteins for international researchers who do not currently have access to Xenopus oocytes. PMID:29530931

The American flag on the VAB is being repainted

NASA Technical Reports Server (NTRS)

1998-01-01

Painters are suspended on platforms from the top of the 525-foot- high Vehicle Assembly Building (VAB) at KSC during repainting of the American flag. The flag spans an area 209 feet by 110 feet and will require 510 gallons of red, white and blue paint. Each stripe of the flag is 9 feet wide and each star is 6 feet in diameter. The platforms are operated by two electric motors and travel 35 feet per minute. Work is being done with rollers, with brushes being used for details. The paint was donated by ICI Devoe of Louisville, Ky. In addition to the flag, the Bicentennial Emblem on the other side of the VAB doors is being replaced by the NASA logo, honoring NASA's 40th anniversary (in October). The logo covers an area 110 feet by 132 feet. Work is expected to be completed in mid-September.

KSC-2011-1487

NASA Image and Video Library

2011-02-18

CAPE CANAVERAL, Fla. -- Members of the United States Air Force 45th Space Wing Honor Guard and more than a dozen 9/11 first responders take part in "The National 9/11 Flag" stitching ceremony in the Debus Conference Facility at the Kennedy Space Center Visitor Complex in Florida. The contributions of NASA, Kennedy Space Center and the state of Florida were stitched into the fabric of the American Flag, which was recovered near ground zero following the World Trade Center attacks on Sept. 11, 2001. The "New York Says Thank You Foundation" is taking the flag on a cross-country journey to be restored to its original 13-stripe design using pieces of fabric from American flags destined for retirement in all 50 states. Once the flag is restored, it will become a permanent collection of the National September 11 Memorial Museum being built at the World Trade Center site. Photo credit: NASA/Kim Shiflett

KSC-2011-1492

NASA Image and Video Library

2011-02-18

CAPE CANAVERAL, Fla. – Deputy Fire Chief Rick Anderson, left, Chief of Fire Training George Hoggard, and Assistant Chief of Fire Training David Seymour with NASA Kennedy Space Center Protective Services participated in the "National 9/11 Flag" stitching ceremony in the Debus Conference Facility at the Kennedy Space Center Visitor Complex in Florida. The contributions of NASA, Kennedy Space Center and the state of Florida were stitched into the fabric of the American Flag, which was recovered near ground zero following the World Trade Center attacks on Sept. 11, 2001. The "New York Says Thank You Foundation" is taking the flag on a cross-country journey to be restored to its original 13-stripe design using pieces of fabric from American flags destined for retirement in all 50 states. Once the flag is restored, it will become a permanent collection of the National September 11 Memorial Museum being built at the World Trade Center site. Photo credit: NASA/Kim Shiflett

KSC-2011-1488

NASA Image and Video Library

2011-02-18

CAPE CANAVERAL, Fla. -- In the Debus Conference Facility at the Kennedy Space Center Visitor Complex in Florida, Jeff Parness, the director, founder and chairman of the "New York Says Thank You Foundation" talks about the work and devotion that has gone into restoring "The National 9/11 Flag." The contributions of NASA, Kennedy Space Center and the state of Florida were stitched into the fabric of the American Flag, which was recovered near ground zero following the World Trade Center attacks on Sept. 11, 2001. The "New York Says Thank You Foundation" is taking the flag on a cross-country journey to be restored to its original 13-stripe design using pieces of fabric from American flags destined for retirement in all 50 states. Once the flag is restored, it will become a permanent collection of the National September 11 Memorial Museum being built at the World Trade Center site. Photo credit: NASA/Kim Shiflett

The Savannah River Site`s Groundwater Monitoring Program, first quarter 1989

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

Not Available

The Environmental Monitoring Section of the Environmental and Health Protection (EHP) Department administers the Savannah River Site`s Groundwater Monitoring Program. During first quarter 1989 (January--March), EHP conducted routine sampling of monitoring wells and drinking water locations. EHP collected the drinking water samples from Savannah River Site (SRS) drinking water systems supplied by wells. EHP established two sets of flagging criteria in 1986 to assist in the management of sample results. The flagging criteria aid personnel in sample scheduling, interpretation of data, and trend identification. The flagging criteria are based on detection limits, background levels in SRS groundwater, and drinking watermore » standards. An explanation of flagging criteria for the first quarter is presented in the Flagging Criteria section of this document. All analytical results from first quarter 1989 are listed in this report, which is distributed to all waste-site custodians.« less

The Savannah River Site`s Groundwater Monitoring Program, third quarter 1989

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

Not Available

The Environmental Monitoring Section of the Environmental and Health Protection (EHP) Department administers the Savannah River Site`s Groundwater Monitoring Program. During third quarter 1989 (July--September), EHP conducted routine sampling of monitoring wells and drinking water locations. EHP collected the drinking water samples from Savannah River Site (SRS) drinking water systems supplied by wells. EHP established two sets of flagging criteria in 1986 to assist in the management of sample results. The flagging criteria do not define contamination levels; instead they aid personnel in sample scheduling, interpretation of data, and trend identification. The flagging criteria are based on detection limits, backgroundmore » levels in SRS groundwater, and drinking water standards. An explanation of flagging criteria for the third quarter is presented in the Flagging Criteria section of this document. All analytical results from third quarter 1989 are listed in this report, which is distributed to all waste-site custodians.« less

The Savannah River Site's Groundwater Monitoring Program, third quarter 1989

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

Not Available

The Environmental Monitoring Section of the Environmental and Health Protection (EHP) Department administers the Savannah River Site's Groundwater Monitoring Program. During third quarter 1989 (July--September), EHP conducted routine sampling of monitoring wells and drinking water locations. EHP collected the drinking water samples from Savannah River Site (SRS) drinking water systems supplied by wells. EHP established two sets of flagging criteria in 1986 to assist in the management of sample results. The flagging criteria do not define contamination levels; instead they aid personnel in sample scheduling, interpretation of data, and trend identification. The flagging criteria are based on detection limits, backgroundmore » levels in SRS groundwater, and drinking water standards. An explanation of flagging criteria for the third quarter is presented in the Flagging Criteria section of this document. All analytical results from third quarter 1989 are listed in this report, which is distributed to all waste-site custodians.« less

The Savannah River Site's Groundwater Monitoring Program, first quarter 1989

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

Not Available

The Environmental Monitoring Section of the Environmental and Health Protection (EHP) Department administers the Savannah River Site's Groundwater Monitoring Program. During first quarter 1989 (January--March), EHP conducted routine sampling of monitoring wells and drinking water locations. EHP collected the drinking water samples from Savannah River Site (SRS) drinking water systems supplied by wells. EHP established two sets of flagging criteria in 1986 to assist in the management of sample results. The flagging criteria aid personnel in sample scheduling, interpretation of data, and trend identification. The flagging criteria are based on detection limits, background levels in SRS groundwater, and drinking watermore » standards. An explanation of flagging criteria for the first quarter is presented in the Flagging Criteria section of this document. All analytical results from first quarter 1989 are listed in this report, which is distributed to all waste-site custodians.« less

Identification of Short Hairpin RNA Targeting Foot-And-Mouth Disease Virus with Transgenic Bovine Fetal Epithelium Cells

PubMed Central

He, Hongbin; Ding, Fangrong; Yang, Hongjun; Cheng, Lei; Liu, Wenhao; Zhong, Jifeng; Dai, Yunping; Li, Guangpeng; He, Chengqiang; Yu, Li; Li, Jianbin

2012-01-01

Background Although it is known that RNA interference (RNAi) targeting viral genes protects experimental animals, such as mice, from the challenge of Foot-and-mouth disease virus (FMDV), it has not been previously investigated whether shRNAs targeting FMDV in transgenic dairy cattle or primary transgenic bovine epithelium cells will confer resistance against FMDV challenge. Principal Finding Here we constructed three recombinant lentiviral vectors containing shRNA against VP2 (RNAi-VP2), VP3 (RNAi-VP3), or VP4 (RNAi-VP4) of FMDV, and found that all of them strongly suppressed the transient expression of a FLAG-tagged viral gene fusion protein in 293T cells. In BHK-21 cells, RNAi-VP4 was found to be more potent in inhibition of viral replication than the others with over 98% inhibition of viral replication. Therefore, recombinant lentiviral vector RNAi-VP4 was transfected into bovine fetal fibroblast cells to generate transgenic nuclear donor cells. With subsequent somatic cell cloning, we generated forty transgenic blastocysts, and then transferred them to 20 synchronized recipient cows. Three transgenic bovine fetuses were obtained after pregnant period of 4 months, and integration into chromosome in cloned fetuses was confirmed by Southern hybridization. The primary tongue epithelium cells of transgenic fetuses were isolated and inoculated with 100 TCID50 of FMDV, and it was observed that shRNA significantly suppressed viral RNA synthesis and inhibited over 91% of viral replication after inoculation of FMDV for 48 h. Conclusion RNAi-VP4 targeting viral VP4 gene appears to prevent primary epithelium cells of transgenic bovine fetus from FMDV infection, and it could be a candidate shRNA used for cultivation of transgenic cattle against FMDV. PMID:22905125

Useful information provided by graphic displays of automated cell counter in hematological malignancies.

PubMed

Gupta, Monica; Chauhan, Kriti; Singhvi, Tanvi; Kumari, Manisha; Grover, Rajesh Kumar

2018-01-21

Automated cell counters have become more and more sophisticated with passing years. The numerical and graphic data both provide useful clues for suspecting a diagnosis especially when the workload is very high. We present our experience of useful information provided by graphic displays of an automated cell counter in hematological malignancies in a cancer hospital where a large number of complete blood count (CBC) requests are received either before or during chemotherapy. This study was conducted to assess the usefulness of hematology cell counter, viz. WBC-Diff (WBC differential), WBC/BASO (WBC basophil) and IMI (immature myeloid information) channel scatter plots, and the flaggings generated in various hematological malignancies. The graphic displays have been compiled over a period of 1 year (October 2015-September 2016) from blood samples of various solid and hematological malignancies (approximately 400 per day) received for routine CBC in the laboratory. Approximately 50 000 scattergrams have been analyzed during the study period. The findings were confirmed by peripheral blood smear examination. The scattergram analysis on XE-2100 is very sensitive as well as specific for diagnosing acute leukemia, viz. acute myeloid leukemia, acute lymphoblastic leukemia; chronic myeloproliferative disorders, viz. chronic myeloid leukemia; and chronic lymphoproliferative disorder especially chronic lymphocytic leukemia. It is suggested that the laboratories using the hematology analyzers be aware of graphic display patterns in addition to flaggings generated which provide additional information and give clue toward the diagnosis even before peripheral smear examination. © 2018 Wiley Periodicals, Inc.

Identification of dually acylated proteins from complementary DNA resources by cell-free and cellular metabolic labeling.

PubMed

Moriya, Koko; Kimoto, Mayumi; Matsuzaki, Kanako; Kiwado, Aya; Takamitsu, Emi; Utsumi, Toshihiko

2016-10-15

To establish a strategy to identify dually fatty acylated proteins from cDNA resources, seven N-myristoylated proteins with cysteine (Cys) residues within the 10 N-terminal residues were selected as potential candidates among 27 N-myristoylated proteins identified from a model human cDNA resource. Seven proteins C-terminally tagged with FLAG tag or EGFP were generated and their susceptibility to protein N-myristoylation and S-palmitoylation were evaluated by metabolic labeling with [(3)H]myristic acid or [(3)H]palmitic acid either in an insect cell-free protein synthesis system or in transfected mammalian cells. As a result, EEPD1, one of five proteins (RFTN1, EEPD1, GNAI1, PDE2A, RNF11) found to be dually acylated, was shown to be a novel dually fatty acylated protein. Metabolic labeling experiments using G2A and C7S mutants of EEPD1-EGFP revealed that the palmitoylation site of EEPD1 is Cys at position 7. Analysis of the intracellular localization of EEPD1 C-terminally tagged with FLAG tag or EGFP and its G2A and C7S mutants revealed that the dual acylation directs EEPD1 to localize to the plasma membrane. Thus, dually fatty acylated proteins can be identified from cDNA resources by cell-free and cellular metabolic labeling of N-myristoylated proteins with Cys residue(s) close to the N-myristoylated N-terminus. Copyright © 2016 Elsevier Inc. All rights reserved.

77 FR 29438 - Self-Regulatory Organizations; EDGX Exchange, Inc.; Notice of Filing and Immediate Effectiveness...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-05-17

... ``added liquidity'' to ``removed liquidity'' ratio of at least 70% where added flags are defined as B, HA..., Flag N is yielded when an order removes liquidity from the EDGX book in Tapes B or C securities. In... Members, the Exchange proposes to amend Flag N so that it only applies to orders that remove liquidity...

No Global Citizenship? Re-Envisioning Global Citizenship Education in Times of Growing Nationalism

ERIC Educational Resources Information Center

Barrow, Elizabeth

2017-01-01

This article presents a discussion of the author's concern over a statement President Trump made in his first Thank You Tour speech, given Dec 1, 2016, in Cincinnati Ohio. "There is no global anthem. No global currency. No certificate of global citizenship. We pledge allegiance to one flag and that flag is the American flag." Here…

Know Your America: Suggested Study Course in Americanism. Revised Edition.

ERIC Educational Resources Information Center

American Legion, Indianapolis, IN. Americanism and Children's Youth Div.

The purpose of this booklet is to increase understanding of fundamental U.S. documents, the U.S. flag, patriotic institutions, and of San Francisco (California), March 14-16, 1986 of U.S. residents. Unit 2 describes and interprets the code of displaying the U.S. flag and provides a suggested flag education unit of study. Units 3 and 4 offer…

A Retrospective Estimate of Ear Disease Detection Using the "Red Flags" in a Clinical Sample.

PubMed

Klyn, Niall A M; Kleindienst Robler, Samantha; Alfakir, Razan; Nielsen, Donald W; Griffith, James W; Carlson, Deborah L; Lundy, Larry; Dhar, Sumitrajit; Zapala, David A

2018-03-01

The purpose of this study was to evaluate the specificity and sensitivity of two red flag protocols in detecting ear diseases associated with changes in hearing. The presence of red-flag symptoms was determined in a chart review of 307 adult patients from the Mayo Clinic Florida Departments of Otorhinolaryngology and Audiology. Participants formed a convenience sample recruited for a separate study. Neurotologist diagnosis was the criterion for comparisons. Of the 251 patient files retained for analysis, 191 had one or more targeted diseases and 60 had age- or noise-related hearing loss. Food and Drug Administration red flags sensitivity was 91% (confidence interval [CI], 86 to 95%) and specificity was 72% (CI, 59 to 83%). American Academy of Otolaryngology-Head and Neck Surgery red flags sensitivity was 98% (CI, 95 to 99%) and specificity was 20% (CI, 11 to 32%). Stakeholders must determine which diseases are meaningful contraindications for hearing aid use and whether these red-flag protocols have acceptable levels of sensitivity and specificity. As direct-to-consumer models of hearing devices increase, a disease detection method that does not require provider intercession would be useful.

Biological properties of purified recombinant HCV particles with an epitope-tagged envelope

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

Takahashi, Hitoshi; Akazawa, Daisuke; Toray Industries, Inc., Kanagawa

2010-05-14

To establish a simple system for purification of recombinant infectious hepatitis C virus (HCV) particles, we designed a chimeric J6/JFH-1 virus with a FLAG (FL)-epitope-tagged sequence at the N-terminal region of the E2 hypervariable region-1 (HVR1) gene (J6/JFH-1/1FL). We found that introduction of an adaptive mutation at the potential N-glycosylation site (E2N151K) leads to efficient production of the chimeric virus. This finding suggests the involvement of glycosylation at Asn within the envelope protein(s) in HCV morphogenesis. To further analyze the biological properties of the purified recombinant HCV particles, we developed a strategy for large-scale production and purification of recombinant J6/JFH-1/1FL/E2N151K.more » Infectious particles were purified from the culture medium of J6/JFH-1/1FL/E2N151K-infected Huh-7 cells using anti-FLAG affinity chromatography in combination with ultrafiltration. Electron microscopy of the purified particles using negative staining showed spherical particle structures with a diameter of 40-60 nm and spike-like projections. Purified HCV particle-immunization induced both an anti-E2 and an anti-FLAG antibody response in immunized mice. This strategy may contribute to future detailed analysis of HCV particle structure and to HCV vaccine development.« less

Three endoplasmic reticulum-associated fatty acyl-coenzyme a reductases were involved in the production of primary alcohols in hexaploid wheat (Triticum aestivum L.).

PubMed

Chai, Guaiqiang; Li, Chunlian; Xu, Feng; Li, Yang; Shi, Xue; Wang, Yong; Wang, Zhonghua

2018-03-05

The cuticle covers the surface of the polysaccharide cell wall of leaf epidermal cells and forms an essential diffusion barrier between the plant and the environment. The cuticle is composed of cutin and wax. Cuticular wax plays an important role in the survival of plants by serving as the interface between plants and their biotic and abiotic environments, especially restricting nonstomatal water loss. Leaf cuticular waxes of hexaploid wheat at the seedling stage mainly consist of primary alcohols, aldehydes, fatty acids, alkane and esters. Primary alcohols account for more than 80% of the total wax load. Therefore, we cloned several genes encoding fatty acyl-coenzyme A reductases from wheat and analyzed their function in yeast and plants. We propose the potential use of these genes in wheat genetic breeding. We reported the cloning and characterization of three TaFARs, namely TaFAR6, TaFAR7 and TaFAR8, encoding fatty acyl-coenzyme A reductases (FAR) in wheat leaf cuticle. Expression analysis revealed that TaFAR6, TaFAR7 and TaFAR8 were expressed at the higher levels in the seedling leaf blades, and were expressed moderately or weakly in stamen, glumes, peduncle, flag leaf blade, sheath, spike, and pistil. The heterologous expression of three TaFARs in yeast (Saccharomyces cerevisiae) led to the production of C24:0 and C26:0 primary alcohols. Transgenic expression of the three TaFARs in tomato (Solanum lycopersicum) and rice (Oryza sativa) led to increased accumulation of C24:0-C30:0 primary alcohols. Transient expression of GFP protein-tagged TaFARs revealed that the three TaFAR proteins were localized to the endoplasmic reticulum (ER), the site of wax biosynthesis. The three TaFAR genes were transcriptionally induced by drought, cold, heat, powdery mildew (Blumeria graminis) infection, abscisic acid (ABA) and methyl jasmonate (MeJa) treatments. These results indicated that wheat TaFAR6, TaFAR7 and TaFAR8 are involved in biosynthesis of very-long-chain primary alcohols in hexaploid wheat and in response to multiple environmental stresses.

HBpF-proBDNF: A New Tool for the Analysis of Pro-Brain Derived Neurotrophic Factor Receptor Signaling and Cell Biology.

PubMed

Gaub, Perrine; de Léon, Andrès; Gibon, Julien; Soubannier, Vincent; Dorval, Geneviève; Séguéla, Philippe; Barker, Philip A

2016-01-01

Neurotrophins activate intracellular signaling pathways necessary for neuronal survival, growth and apoptosis. The most abundant neurotrophin in the adult brain, brain-derived neurotrophic factor (BDNF), is first synthesized as a proBDNF precursor and recent studies have demonstrated that proBDNF can be secreted and that it functions as a ligand for a receptor complex containing p75NTR and sortilin. Activation of proBDNF receptors mediates growth cone collapse, reduces synaptic activity, and facilitates developmental apoptosis of motoneurons but the precise signaling cascades have been difficult to discern. To address this, we have engineered, expressed and purified HBpF-proBDNF, an expression construct containing a 6X-HIS tag, a biotin acceptor peptide (BAP) sequence, a PreScission™ Protease cleavage site and a FLAG-tag attached to the N-terminal part of murine proBDNF. Intact HBpF-proBDNF has activities indistinguishable from its wild-type counterpart and can be used to purify proBDNF signaling complexes or to monitor proBDNF endocytosis and retrograde transport. HBpF-proBDNF will be useful for characterizing proBDNF signaling complexes and for deciphering the role of proBDNF in neuronal development, synapse function and neurodegenerative disease.

Microbeads display of proteins using emulsion PCR and cell-free protein synthesis.

PubMed

Gan, Rui; Yamanaka, Yumiko; Kojima, Takaaki; Nakano, Hideo

2008-01-01

We developed a method for coupling protein to its coding DNA on magnetic microbeads using emulsion PCR and cell-free protein synthesis in emulsion. A PCR mixture containing streptavidin-coated microbeads was compartmentalized by water-in-oil (w/o) emulsion with estimated 0.5 template molecules per droplet. The template molecules were amplified and immobilized on beads via bead-linked reverse primers and biotinylated forward primers. After amplification, the templates were sequentially labeled with streptavidin and biotinylated anti-glutathione S-transferase (GST) antibody. The pool of beads was then subjected to cell-free protein synthesis compartmentalized in another w/o emulsion, in which templates were coupled to their coding proteins. We mixed two types of DNA templates of Histidine6 tag (His6)-fused and FLAG tag-fused GST in a ratio of 1:1,000 (His6: FLAG) for use as a model DNA library. After incubation with fluorescein isothiocyanate (FITC)-labeled anti-His6 (C-term) antibody, the beads with the His6 gene were enriched 917-fold in a single-round screening by using flow cytometry. A library with a theoretical diversity of 10(6) was constructed by randomizing the middle four residues of the His6 tag. After a two-round screening, the randomized sequences were substantially converged to peptide-encoding sequences recognized by the anti-His6 antibody.

Distributed Password Cracking

DTIC Science & Technology

2009-12-01

other services for early UNIX systems at Bell labs. In many UNIX based systems, the field added to ‘etc/ passwd ’ file to carry GCOS ID information was...charset, and external. struct options_main { /* Option flags */ opt_flags flags; /* Password files */ struct list_main * passwd ; /* Password file...object PASSWD . It is part of several other data structures. struct PASSWD { int id; char *login; char *passwd_hash; int UID

Cell cycle accumulation of the proliferating cell nuclear antigen PCN-1 transitions from continuous in the adult germline to intermittent in the early embryo of C. elegans.

PubMed

Kocsisova, Zuzana; Kornfeld, Kerry; Schedl, Tim

2018-05-30

The proliferating cell nuclear antigen (PCNA or PCN-1 in C. elegans), an essential processivity factor for DNA polymerase δ, has been widely used as a marker of S-phase. In C. elegans early embryos, PCN-1 accumulation is cyclic, localizing to the nucleus during S-phase and the cytoplasm during the rest of the cell cycle. The C. elegans larval and adult germline is an important model systems for studying cell cycle regulation, and it was observed that the cell cycle regulator cyclin E (CYE-1 in C. elegans) displays a non-cyclic, continuous accumulation pattern in this tissue. The accumulation pattern of PCN-1 has not been well defined in the larval and adult germline, and the objective of this study was to determine if the accumulation pattern is cyclic, as in other cells and organisms, or continuous, similar to cyclin E. To study the larval and adult germline accumulation of PCN-1 expressed from its native locus, we used CRISPR/Cas9 technology to engineer a novel allele of pcn-1 that encodes an epitope-tagged protein. S-phase nuclei were labeled using EdU nucleotide incorporation, and FLAG::PCN-1 was detected by antibody staining. All progenitor zone nuclei, including those that were not in S-phase (as they were negative for EdU staining) showed PCN-1 accumulation, indicating that PCN-1 accumulated during all cell cycle phases in the germline progenitor zone. The same result was observed with a GFP::PCN-1 fusion protein expressed from a transgene. pcn-1 loss-of-function mutations were analyzed, and pcn-1 was necessary for robust fertility and embryonic development. In the C. elegans early embryo as well as other organisms, PCN-1 accumulates in nuclei only during S-phase. By contrast, in the progenitor zone of the germline of C. elegans, PCN-1 accumulated in nuclei during all cell cycle stages. This pattern is similar to accumulation pattern of cyclin E. These observations support the model that mitotic cell cycle regulation in the germline stem and progenitor cells is distinct from somatic cells, as it does not heavily rely on cyclic accumulation of classic cell cycle proteins.

KSC-07pd0924

NASA Image and Video Library

2007-04-19

KENNEDY SPACE CENTER, FLA. -- The finishing touches are painted on the American flag that embellishes the southwest side of the Vehicle Assembly Building at NASA's Kennedy Space Center. The flag and the NASA logo, which is on the southeast side, have both been refreshed with new paint. The flag and logo were last painted in 1998, honoring NASA's 40th anniversary. The flag spans an area 209 feet by 110 feet, or about 23, 437 square feet. Each stripe is 9 feet wide and each star is 6 feet in diameter. The logo, which is known as the "meatball," measures 110 feet by 132 feet, or about 12,300 square feet. The building stands 525-feet tall. Photo credit: NASA/George Shelton

An Auto-flag Method of Radio Visibility Data Based on Support Vector Machine

NASA Astrophysics Data System (ADS)

Dai, Hui-mei; Mei, Ying; Wang, Wei; Deng, Hui; Wang, Feng

2017-01-01

The Mingantu Ultrawide Spectral Radioheliograph (MUSER) has entered a test observation stage. After the construction of the data acquisition and storage system, it is urgent to automatically flag and eliminate the abnormal visibility data so as to improve the imaging quality. In this paper, according to the observational records, we create a credible visibility set, and further obtain the corresponding flag model of visibility data by using the support vector machine (SVM) technique. The results show that the SVM is a robust approach to flag the MUSER visibility data, and can attain an accuracy of about 86%. Meanwhile, this method will not be affected by solar activities, such as flare eruptions.

Flagging versus dragging as sampling methods for nymphal Ixodes scapularis (Acari: Ixodidae)

USGS Publications Warehouse

Rulison, Eric L.; Kuczaj, Isis; Pang, Genevieve; Hickling, Graham J.; Tsao, Jean I.; Ginsberg, Howard S.

2013-01-01

The nymphal stage of the blacklegged tick, Ixodes scapularis (Acari: Ixodidae), is responsible for most transmission of Borrelia burgdorferi, the etiologic agent of Lyme disease, to humans in North America. From 2010 to fall of 2012, we compared two commonly used techniques, flagging and dragging, as sampling methods for nymphal I. scapularis at three sites, each with multiple sampling arrays (grids), in the eastern and central United States. Flagging and dragging collected comparable numbers of nymphs, with no consistent differences between methods. Dragging collected more nymphs than flagging in some samples, but these differences were not consistent among sites or sampling years. The ratio of nymphs collected by flagging vs dragging was not significantly related to shrub density, so habitat type did not have a strong effect on the relative efficacy of these methods. Therefore, although dragging collected more ticks in a few cases, the numbers collected by each method were so variable that neither technique had a clear advantage for sampling nymphal I. scapularis.

Molecular Studies on MIC1/PDF in Human Prostate Cancer

DTIC Science & Technology

2005-09-01

expression of secreted MIC-1 protein (Aim 2). Moreover, the functions of MIC-1 have been studied in vitro by using specific antibody directed against MIC-1... antibody M2 (Sigma). The FLAG tag was inserted at the 3’-translated region of the MIC-l/PDF cDNA. A double stranded, synthetic oligonucleotide was designed...B) Lower panel showed the coomassie blue of specific antibody directed against stained gel as a loading control. MIC-1 protein. More specifically

Targeting FASN for Breast Cancer Treatment by Repositioning PPIs

DTIC Science & Technology

2017-01-01

the sole cytosolic enzyme responsible for de novo synthesis of palmitate. Recently, it was found that FASN up-regulation contributes to drug and...increased whereas Flag-p65 (NF-κB) over-expression significantly reduced PARP-1 protein and mRNA levels. Knocking down p65 using shRNA significantly...increased PARP-1 protein and mRNA levels (Fig. 2C), whereas activation of p65 with TNF-α had contrary effect (Fig. 2D). Taken together, these results

78 FR 966 - Self-Regulatory Organizations; EDGX Exchange, Inc.; Notice of Filing and Immediate Effectiveness...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-01-07

... the HP-API order entry protocol (HP-API) in order to qualify for the rates on Flags ZA and ZR. The... via FIX in order to qualify for the rates on Flags ZA (rebate of $0.0032 per share) and ZR (fee of $0... qualify for the rates on Flags ZA and ZR. The attestation requirement, as described above and in SR-EDGX...

77 FR 61797 - Self-Regulatory Organizations; EDGX Exchange, Inc.; Notice of Filing and Immediate Effectiveness...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-10-11

..., MM, RP, 3, or 4 and removal flags are defined as Flags BB, MT, N, W, PI, PR, or 6. Where a Member..., BB and PI where they satisfy the volume tier requirements for the Mega Tier in Footnote 1. Currently... removing liquidity and the rate for Flags N, W, 6, BB, and PI subject to the volume thresholds in Footnotes...

The Latest Results from the Focal L-Band Array for the Green Bank Telescope (FLAG), the World's (Current) Most Sensitive Phased Array Feed

NASA Astrophysics Data System (ADS)

Pingel, Nickolas; Pisano, D. J.

2018-01-01

Phased Array Feeds (PAFs) represent the next revolution in radio astronomy instrumentation. I will present results from the latest commissioning run from the Focal L-Band Array for the Green Bank telescope (FLAG), which holds the current world record for PAF sensitivity. Since we are able to operate at system temperatures comparable with the traditional GBT single pixel L-Band feed, the increase in the field-of-view provided by the beamforming capabilities of PAFs results in a dramatic (a factor of 5) increase in survey speeds. In particular, FLAG can probe similar neutral hydrogen column density regimes over a 4 sq. deg region in 24.6 minutes as opposed to 4.1 hours in an equivalent single pixel map (excluding observing overhead). In addition to comparisons between data taken with FLAG and the single-pixel L-Band feed, I will also discuss the technical aspects of the observing procedure, data reduction, and the transition path for FLAG from an instrument that is principle-investigator run to one that is general use. These FLAG results provide a very encouraging outlook on how the GBT will continue to compete with current and planned radio telescope facilities.

Red eyes and red-flags: improving ophthalmic assessment and referral in primary care.

PubMed

Kilduff, Caroline; Lois, Charis

2016-01-01

Up to five percent of primary care consultations are eye-related, yet 96% of General Practitioners (GPs) do not undergo postgraduate ophthalmology training. Most do not feel assured performing eye assessments. Some red eye conditions can become sight threatening, and often exhibit red-flag features. These features include moderate pain, photophobia, reduced visual acuity (VA), eye-trauma, or unilateral marked redness. The aim of this project was to improve primary care assessment and referral of patients presenting with red-flag features based on the NICE 'Red Eye' Clinical Knowledge Summary recommendations. Data was collected retrospectively from 139 red eye consultations. A practice meeting highlighted poor awareness of red-flag features, low confidence levels in eye assessments, and time-constraints during appointments. Interventions were based on feedback from staff. These included a primary care teaching session on red-flag features, a VA measurement tutorial, and provision of a red eye toolkit, including VA equipment, to each consultation room. At baseline, each patient had on average 0.9 red-flag features assessed. Only 36.0% (9/25) of patients with red-flag features were appropriately referred to same-day ophthalmology services. Following two improvement cycles, a significant improvement was seen in almost every parameter. On average, each patient had 2.7 red-flag features assessed (vs 0.9, p<0.001). VA was assessed in 55.6% of consultations (vs 7.9%, p<0.001), pain was quantified in 81.5% (vs 20.9%, p=0.005), eye-trauma or foreign-body (51.8% vs 8.6%, p<0.001), extent of redness was documented in 66.7% (vs 14.4%, p<0.001). Only photophobia remained poorly assessed (18.5% vs 14.4%, p=0.75). Following this, 75.0% (6/8) of patients were appropriately referred. This project reflected the literature regarding low confidence and inexperience amongst GPs when faced with ophthalmic conditions. Improvements in education are required to ensure accurate assessments can be undertaken in a time-constrained environment.

Rome III survey of irritable bowel syndrome among ethnic Malays

PubMed Central

Lee, Yeong Yeh; Waid, Anuar; Tan, Huck Joo; Chua, Andrew Seng Boon; Whitehead, William E

2012-01-01

AIM: To survey irritable bowel syndrome (IBS) using Rome III criteria among Malays from the north-eastern region of Peninsular Malaysia. METHODS: A previously validated Malay language Rome III IBS diagnostic questionnaire was used in the current study. A prospective sample of 232 Malay subjects (80% power) was initially screened. Using a stratified random sampling strategy, a total of 221 Malay subjects (112 subjects in a “full time job” and 109 subjects in “no full time job”) were recruited. Subjects were visitors (friends and relatives) within the hospital compound and were representative of the local community. Red flags and psychosocial alarm symptoms were also assessed in the current study using previously translated and validated questionnaires. Subjects with IBS were sub-typed into constipation-predominant, diarrhea-predominant, mixed type and un-subtyped. Univariable and multivariable analyses were used to test for association between socioeconomic factors and presence of red flags and psychosocial alarm features among the Malays with IBS. RESULTS: IBS was present in 10.9% (24/221), red flags in 22.2% (49/221) and psychosocial alarm features in 9.0% (20/221). Red flags were more commonly reported in subjects with IBS (83.3%) than psychosocial alarm features (20.8%, P < 0.001). Subjects with IBS were older (mean age 41.4 years vs 36.9 years, P = 0.08), but no difference in gender was noted (P = 0.4). Using univariable analysis, IBS was significantly associated with a tertiary education, high individual income above RM1000, married status, ex-smoker and the presence of red flags (all P < 0.05). In multiple logistic regression analysis, only the presence of red flags was significantly associated with IBS (odds ratio: 0.02, 95%CI: 0.004-0.1, P < 0.001). The commonest IBS sub-type was mixed type (58.3%), followed by constipation-predominant (20.8%), diarrhea-predominant (16.7%) and un-subtyped (4.2%). Four of 13 Malay females (30.8%) with IBS also had menstrual pain. Most subjects with IBS had at least one red flag (70.8%), 12.5% had two red flags and 16.7% with no red flags. The commonest red flag was a bowel habit change in subjects > 50 years old and this was reported by 16.7% of subjects with IBS. CONCLUSION: Using the Rome III criteria, IBS was common among ethnic Malays from the north-eastern region of Peninsular Malaysia. PMID:23197894

Pyrimidine tract-binding protein 1 mediates pyruvate kinase M2-dependent phosphorylation of signal transducer and activator of transcription 3 and oncogenesis in anaplastic large cell lymphoma.

PubMed

Hwang, Steven R; Murga-Zamalloa, Carlos; Brown, Noah; Basappa, Johnvesly; McDonnell, Scott Rp; Mendoza-Reinoso, Veronica; Basrur, Venkatesha; Wilcox, Ryan; Elenitoba-Johnson, Kojo; Lim, Megan S

2017-08-01

PKM2 (pyruvate kinase M2), a critical regulator of glycolysis, is phosphorylated by numerous growth factor receptors and oncogenic tyrosine kinases including NPM-ALK which is expressed in a subset of aggressive T-cell non-Hodgkin lymphomas known as anaplastic large cell lymphoma, ALK-positive. Our previous work demonstrated that phosphorylation of Y105-PKM2 by NPM-ALK regulates a major metabolic shift to promote lymphomagenesis. In addition to its role in metabolism, recent studies have shown that PKM2 promotes oncogenesis by phosphorylating nuclear STAT3 (signal transducer and activator of transcription 3) and regulating transcription of genes involved in cell survival and proliferation. We hypothesized that identification of novel PKM2 interactors could provide additional insights into its expanding functional role in cancer. To this end, immunocomplexes of FLAG-tagged PKM2 were isolated from NPM-ALK-positive ALCL (anaplastic large cell lymphoma) cells and subjected to liquid chromatography tandem mass spectrometry (LC-MS/MS) which led to the identification of polypyrimidine tract-binding protein (PTBP1) as a novel interactor of PKM2. The interaction between PTBP1 and PKM2 was restricted to the nucleus and was dependent on NPM-ALK mediated Y105 phosphorylation of PKM2. Stable shRNA-mediated silencing of PTBP1 resulted in a marked decrease in pY105-PKM2 and pY705-STAT3 which led to decreased ALCL cell proliferation and colony formation. Overall, our data demonstrate that PTBP1 interacts with PKM2 and promotes ALCL oncogenesis by facilitating PKM2-dependent activation of STAT3 within the nucleus.

Involvement of Abscisic Acid in PSII Photodamage and D1 Protein Turnover for Light-Induced Premature Senescence of Rice Flag Leaves

PubMed Central

Wang, Fubiao; Liu, Jianchao; Chen, Minxue; Zhou, Lujian; Li, Zhaowei; Zhao, Qian; Pan, Gang; Zaidi, Syed-Hassan-Raza; Cheng, Fangmin

2016-01-01

D1 protein in the PSII reaction center is the major target of photodamage, and it exhibits the highest turnover rate among all the thylakoid proteins. In this paper, rice psf (premature senescence of flag leaves) mutant and its wild type were used to investigate the genotype-dependent alteration in PSII photo-damage and D1 protein turnover during leaf senescence and its relation to ABA accumulation in senescent leaves. The symptom and extent of leaf senescence of the psf mutant appeared to be sunlight-dependent under natural field condition. The psf also displayed significantly higher levels of ABA accumulation in senescent leaves than the wild type. However, the premature senescence lesion of psf leaves could be alleviated by shaded treatment, concomitantly with the strikingly suppressed ABA level in the shaded areas of flag leaves. The change in ABA concentration contributed to the regulation of shade-delayed leaf senescence. The participation of ABA in the timing of senescence initiation and in the subsequent rate of leaf senescence was closely associated with PSII photodamage and D1 protein turnover during leaf senescence, in which the transcriptional expression of several key genes (psbA, psbB, psbC and OsFtsH2) involved in D1 protein biosynthesis and PSII repair cycle was seriously suppressed by the significantly increased ABA level. This response resulted in the low rate of D1 protein synthesis and impaired repair recovery in the presence of ABA. The psf showed evidently decreased D1 protein amount in the senescent leaves. Both the inhibition of de novo synthesized D1 protein and the slow rate of proteolytic removal for the photodamaged D1 protein was among the most crucial steps for the linkage between light-dependent leaf senescence and the varying ABA concentration in psf mutant leaves. OsFtsH2 transcriptional expression possibly played an important role in the regulation of D1 protein turnover and PSII repair cycle in relation to ABA mediated leaf senescence. PMID:27532299

The Lr34 adult plant rust resistance gene provides seedling resistance in durum wheat without senescence.

PubMed

Rinaldo, Amy; Gilbert, Brian; Boni, Rainer; Krattinger, Simon G; Singh, Davinder; Park, Robert F; Lagudah, Evans; Ayliffe, Michael

2017-07-01

The hexaploid wheat (Triticum aestivum) adult plant resistance gene, Lr34/Yr18/Sr57/Pm38/Ltn1, provides broad-spectrum resistance to wheat leaf rust (Lr34), stripe rust (Yr18), stem rust (Sr57) and powdery mildew (Pm38) pathogens, and has remained effective in wheat crops for many decades. The partial resistance provided by this gene is only apparent in adult plants and not effective in field-grown seedlings. Lr34 also causes leaf tip necrosis (Ltn1) in mature adult plant leaves when grown under field conditions. This D genome-encoded bread wheat gene was transferred to tetraploid durum wheat (T. turgidum) cultivar Stewart by transformation. Transgenic durum lines were produced with elevated gene expression levels when compared with the endogenous hexaploid gene. Unlike nontransgenic hexaploid and durum control lines, these transgenic plants showed robust seedling resistance to pathogens causing wheat leaf rust, stripe rust and powdery mildew disease. The effectiveness of seedling resistance against each pathogen correlated with the level of transgene expression. No evidence of accelerated leaf necrosis or up-regulation of senescence gene markers was apparent in these seedlings, suggesting senescence is not required for Lr34 resistance, although leaf tip necrosis occurred in mature plant flag leaves. Several abiotic stress-response genes were up-regulated in these seedlings in the absence of rust infection as previously observed in adult plant flag leaves of hexaploid wheat. Increasing day length significantly increased Lr34 seedling resistance. These data demonstrate that expression of a highly durable, broad-spectrum adult plant resistance gene can be modified to provide seedling resistance in durum wheat. © 2016 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

Blockade of Y177 and Nuclear Translocation of Bcr-Abl Inhibits Proliferation and Promotes Apoptosis in Chronic Myeloid Leukemia Cells.

PubMed

Li, Qianyin; Huang, Zhenglan; Gao, Miao; Cao, Weixi; Xiao, Qin; Luo, Hongwei; Feng, Wenli

2017-03-02

The gradual emerging of resistance to imatinib urgently calls for the development of new therapy for chronic myeloid leukemia (CML). The fusion protein Bcr-Abl, which promotes the malignant transformation of CML cells, is mainly located in the cytoplasm, while the c-Abl protein which is expressed in the nucleus can induce apoptosis. Based on the hetero-dimerization of FKBP (the 12-kDa FK506- and rapamycin-binding protein) and FRB (the FKBP-rapamycin binding domain of the protein kinase, mTOR) mediated by AP21967, we constructed a nuclear transport system to induce cytoplasmic Bcr-Abl into nuclear. In this study, we reported the construction of the nuclear transport system, and we demonstrated that FN3R (three nuclear localization signals were fused to FRBT2098L with a FLAG tag), HF2S (two FKBP domains were in tandem and fused to the SH2 domain of Grb2 with an HA tag) and Bcr-Abl form a complexus upon AP21967. Bcr-Abl was imported into the nucleus successfully by the nuclear transport system. The nuclear transport system inhibited CML cell proliferation through mitogen-activated protein kinase (MAPK) and signal transducer and activator of transcription 5 (STAT5) pathways mainly by HF2S. It was proven that nuclear located Bcr-Abl induced CML cell (including imatinib-resistant K562G01 cells) apoptosis by activation of p73 and its downstream molecules. In summary, our study provides a new targeted therapy for the CML patients even with Tyrosine Kinase Inhibitor (TKI)-resistance.

The Savannah River Site's Groundwater Monitoring Program: Second quarter 1992

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

Rogers, C.D.

1992-10-07

The Environmental Protection Department/Environmental Monitoring Section (EPD/EMS) administers the Savannah River Site's (SRS) Groundwater Monitoring Program. During second quarter 1992, EPD/EMS conducted extensive sampling of monitoring wells. EPD/EMS established two sets of criteria to assist in the management of sample results. The flagging criteria do not define contamination levels; instead, they aid personnel in sample scheduling, interpretation of data, and trend identification. Since 1991, the flagging criteria have been based on the federal Environmental Protection Agency (EPA) drinking water standards and on method detection limits. A detailed explanation of the current flagging criteria is presented in the Flagging Criteria sectionmore » of this document. Analytical results from second quarter 1992 are listed in this report.« less

One isoform of Arg/Abl2 tyrosine kinase is nuclear and the other seven cytosolic isoforms differently modulate cell morphology, motility and the cytoskeleton

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

Bianchi, Cristina; Torsello, Barbara; Di Stefano, Vitalba

The non-receptor tyrosine kinase Abelson related gene (Arg/Abl2) regulates cell migration and morphogenesis by modulating the cytoskeleton. Arg promotes actin-based cell protrusions and spreading, and inhibits cell migration by attenuating stress fiber formation and contractility via activation of the RhoA inhibitor, p190RhoGAP, and by regulating focal adhesion dynamics also via CrkII phosphorylation. Eight full-length Arg isoforms with different N- and C-termini are endogenously expressed in human cells. In this paper, the eight Arg isoforms, subcloned in the pFLAG-CMV2 vector, were transfected in COS-7 cells in order to study their subcellular distribution and role in cell morphology, migration and cytoskeletal modulation.more » The transfected 1BSCTS Arg isoform has a nuclear distribution and phosphorylates CrkII in the nucleus, whilst the other isoforms are detected in the cytoplasm. The 1BLCTL, 1BSCTL, 1ASCTS isoforms were able to significantly decrease stress fibers, induce cell shrinkage and filopodia-like protrusions with a significant increase in p190RhoGAP phosphorylation. In contrast, 1ALCTL, 1ALCTS, 1ASCTL and 1BLCTS isoforms do not significantly decrease stress fibers and induce the formation of retraction tail-like protrusions. The 1BLCTL and 1ALCTL isoforms have different effects on cell migration and focal adhesions. All these data may open new perspectives to study the mechanisms of cell invasiveness. -Highlights: • Each of the eight Arg isoforms was transfected in COS-7 cells. • Only the 1BSCTS Arg isoform has a nuclear distribution in transfected cells. • The cytoplasmic isoforms and F-actin colocalize cortically and in cell protrusions. • Arg isoforms differently phosphorylate p190RhoGAP and CrkII. • Arg isoforms differently modulate stress fibers, cell protrusions and motility.« less

Rolling Band Artifact Flagging in the Kepler Data Pipeline

NASA Astrophysics Data System (ADS)

Clarke, Bruce; Kolodziejczak, Jeffery J; Caldwell, Douglas A.

2014-06-01

Instrument-induced artifacts in the raw Kepler pixel data include time-varying crosstalk from the fine guidance sensor (FGS) clock signals, manifestations of drifting moiré pattern as locally correlated nonstationary noise and rolling bands in the images. These systematics find their way into the calibrated pixel time series and ultimately into the target flux time series. The Kepler pipeline module Dynablack models the FGS crosstalk artifacts using a combination of raw science pixel data, full frame images, reverse-clocked pixel data and ancillary temperature data. The calibration module (CAL) uses the fitted Dynablack models to remove FGS crosstalk artifacts in the calibrated pixels by adjusting the black level correction per cadence. Dynablack also detects and flags spatial regions and time intervals of strong time-varying black-level. These rolling band artifact (RBA) flags are produced on a per row per cadence basis by searching for transit signatures in the Dynablack fit residuals. The Photometric Analysis module (PA) generates per target per cadence data quality flags based on the Dynablack RBA flags. Proposed future work includes using the target data quality flags as a basis for de-weighting in the Presearch Data Conditioning (PDC), Transiting Planet Search (TPS) and Data Validation (DV) pipeline modules. We discuss the effectiveness of RBA flagging for downstream users and illustrate with some affected light curves. We also discuss the implementation of Dynablack in the Kepler data pipeline and present results regarding the improvement in calibrated pixels and the expected improvement in cotrending performance as a result of including FGS corrections in the calibration. Funding for the Kepler Mission has been provided by the NASA Science Mission Directorate.

Challenges to Public Order and the Seas

DTIC Science & Technology

2014-03-01

these excessive claims will ever be rolled back. Worse, they could be strengthened in a game of one- upmanship. A laissez faire approach to flag...to the rule of law and a basis for the conduct of af- fairs among nations. What is necessary for an effective system of ocean governance? This...gain an increased market share as reputable national flags decline. Depending on which FOC is involved, there is a fair probability that the flag state

Divergent receiver responses to components of multimodal signals in two foot-flagging frog species.

PubMed

Preininger, Doris; Boeckle, Markus; Sztatecsny, Marc; Hödl, Walter

2013-01-01

Multimodal communication of acoustic and visual signals serves a vital role in the mating system of anuran amphibians. To understand signal evolution and function in multimodal signal design it is critical to test receiver responses to unimodal signal components versus multimodal composite signals. We investigated two anuran species displaying a conspicuous foot-flagging behavior in addition to or in combination with advertisement calls while announcing their signaling sites to conspecifics. To investigate the conspicuousness of the foot-flagging signals, we measured and compared spectral reflectance of foot webbings of Micrixalus saxicola and Staurois parvus using a spectrophotometer. We performed behavioral field experiments using a model frog including an extendable leg combined with acoustic playbacks to test receiver responses to acoustic, visual and combined audio-visual stimuli. Our results indicated that the foot webbings of S. parvus achieved a 13 times higher contrast against their visual background than feet of M. saxicola. The main response to all experimental stimuli in S. parvus was foot flagging, whereas M. saxicola responded primarily with calls but never foot flagged. Together these across-species differences suggest that in S. parvus foot-flagging behavior is applied as a salient and frequently used communicative signal during agonistic behavior, whereas we propose it constitutes an evolutionary nascent state in ritualization of the current fighting behavior in M. saxicola.

Vortex dynamics and heat transfer behind self-oscillating inverted flags of various lengths in channel flow

NASA Astrophysics Data System (ADS)

Yu, Yuelong; Liu, Yingzheng; Chen, Yujia

2018-04-01

The influence of an inverted flag's length-to-channel-width ratio (C* = L/W) on its oscillating behavior in a channel flow and the resultant vortex dynamics and heat transfer are determined experimentally. Three systems with C* values of 0.125, 0.250, and 0.375 were chosen for comparison. The interaction of highly unsteady flow with the inverted flag is measured with time-resolved particle image velocimetry. Variations in the underlying flow physics are discussed in terms of the statistical flow quantities, flag displacement, phase-averaged flow field, and vortex dynamics. The results show that the increase in C* shifts the occurrence of the flapping regime at high dimensionless bending stiffness. With the flag in the flapping region, three distinct vortex dynamics—the von Kármán vortex street, the G mode, and the singular mode—are identified at C* values of 0.375, 0.250, and 0.125, respectively. Finally, the heat transfer enhancement from the self-oscillating inverted flag is measured to serve as complementary information to quantify the cause-and-effect relationship between vortex dynamics and wall heat transfer. The increase in C* strongly promotes wall heat removal because disruption of the boundary layer by the energetic vortices is substantially intensified. Among all systems, wall heat transfer removal is most efficient at the intermediate C* value of 0.250.

Energy Harvesting for Micropower Applications by Flow-Induced Flutter of an Inverted Piezoelectric Flag

NASA Astrophysics Data System (ADS)

Shoele, Kourosh; Mittal, Rajat

2015-11-01

Piezoelectric flexible flags can be used to continuously generate energy for small-scale sensor used in a wide variety of applications ranging from measurement/monitoring of environmental conditions (outdoors or indoors) to in-situ tracking of wild animals. Here, we study the energy harvesting performance as well as the flow-structure interaction of an inverted piezoelectric flag. We use a coupled fluid-structure-electric solver to examine the dynamic response of the inverted flag as well as the associated vortical characteristics with different inertia and bending stiffness. Simulations indicate that large amplitude vibrations can be achieved over a large range of parameters over which lock-on between the flag flutter and the intrinsic wake shedding occurs. The effects of initial inclination of the flag to the prevailing flow as well as Reynolds number of the flow are explored, and the effect of piezoelectric material parameters on the energy harvesting performance of this flutter state is examined in detail. The maximum energy efficiency occurs when there is a match between the intrinsic timescales of flutter and the piezoelectric circuit. The simulations are used to formulate a scaling law that could be used to predict the energy harvesting performance of such devices. The support for this study comes from AFSOR, NSF, EPRI and Johns Hopkins E2SHI Seed Grant.

Improved Tandem Affinity Purification Tag and Methods for Isolation of Proteins and Protein Complexes from Schizosaccharomyces pombe.

PubMed

Zilio, Nicola; Boddy, Michael N

2017-03-01

The tandem affinity purification (TAP) method uses an epitope that contains two different affinity purification tags separated by a site-specific protease site to isolate a protein rapidly and easily. Proteins purified via the TAP tag are eluted under mild conditions, allowing them to be used for structural and biochemical analyses. The original TAP tag contains a calmodulin-binding peptide and the IgG-binding domain from protein A separated by a tobacco etch virus (TEV) protease cleavage site. After capturing the Protein A epitope on an IgG resin, bound proteins are released by incubation with the TEV protease and then isolated on a calmodulin matrix in the presence of calcium; elution from this resin is achieved by chelating calcium with EGTA. However, because the robustness of the calmodulin-binding step in this procedure is highly variable, we replaced the calmodulin-binding peptide with three copies of the FLAG epitope, (3× FLAG)-TEV-Protein A, which can be isolated using an anti-FLAG resin. Elution from this matrix is achieved in the presence of an excess of a 3× FLAG peptide. In addition to allowing proteins to be released under mild conditions, elution by the 3× FLAG peptide adds an extra layer of specificity to the TAP procedure, because it liberates only FLAG-tagged proteins. © 2017 Cold Spring Harbor Laboratory Press.

49 CFR 234.5 - Definitions.

Code of Federal Regulations, 2011 CFR

2011-10-01

... a vest, shirt, or jacket of a color appropriate for daytime flagging such as orange, yellow, strong.... For nighttime flagging, similar outside garments shall be retro reflective. Acceptable hand signal...

[EFFECT OF Akt1 GENE TRANSFECTION ON HYPOXIA TOLERANCE OF BONE MARROW MESENCHYMAL STEM CELLS].

PubMed

Yu, Fengxu; Chen, Yongen; Chen, Feng; Xia, Jiyi; Liu, Hongduan; Fu, Yong; Li, Miaoling; Liao, Bin

2016-04-01

To investigate whether Akt1 gene transfection mediated by recombinant lentivirus (LVs) in the bone marrow mesenchymal stem cells (BMSCs) could enhance the ability of hypoxia tolerance so as to provide a theoretical basis for improving the effectiveness of stem cells transplantation. LVs was used as transfection vector, enhanced green fluorescent protein (EGFP) was used as markers to construct the pLVX-EGFP-3FLAG virus vector carrying the Akt1 gene. The 3rd generation BMSCs from 3-5 weeks old Sprague Dawley rats were transfected with pLVX-EGFP virus solution as group B and with pLVX-EGFP-3PLAG virus solution as group C; and untransfected BMSCs served as control group (group A). At 2-3 days after transfection, the expression of green fluorescent was observed by fluorescence microscope; and at 48 hours after transfection, Western blot method was used to detect the expression of Akt1 protein in groups B and C. BMSCs of groups B and C were given hypoxia intervention with 94% N₂, 1% O₂, and 5% CO₂ for 0, 3, 6, 9, and 12 hours (group B1 and group C1). The flow cytometry was used to analyze the cell apoptosis rate and cell death rate, and the MTT method to analyze the cell proliferation, and Western blot to detect the expression of apoptosis related gene Caspase-3. After transfection, obvious green fluorescence was observed in BMSCs under fluorescence microscopy in groups B and C, the transfection efficiency was about 60%. Akt1 expression of group C was significantly higher than that of group B (t = 17.525, P = 0.013). The apoptosis rate and cell death rate of group B1 increased gradually with time, and difference was significant (P < 0.05). In group C1, the apoptosis rate and cell death rate decreased temporarily at 3 hours after hypoxia intervention, then increased gradually, and difference was significant (P < 0.05). The apoptosis rate and cell death rate of group C1 were significantly lower than those of group B1 at each time point (P < 0.05) except at 0 hour. MTT assay showed tat absorbance (A) values of groups B and C were significantly higher than those of groups B1 and C1 at each time point (P < 0.05); the A value of group B was significantly lower than that of group C at each time point (P < 0.05). The A value of group B1 was significantly lower than that of group Cl at 6, 9, and 12 hours after hypoxia intervention (P < 0.05). Western blot results showed that the Caspase-3 expression of group C1 significantly reduced when compared with group B1 at each time point (P < 0.05). Akt1 gene transfection mediated by recombinant LVs could significantly improve hypoxia tolerance of BMSCs by inhibiting the apoptosis, which could provide new ideas for improving the effectiveness of stem cells transplantation.

The Savannah River Site's Groundwater Monitoring Program

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

Not Available

1989-01-01

The Environmental Monitoring Section of the Environmental and Health Protection (EHP) Department administers the Savannah River Site's Groundwater Monitoring Program. During fourth quarter 1989 (October--December), EHP conducted routine sampling of monitoring wells and drinking water locations. EHP collected the drinking water samples from Savannah River Site (SRS) drinking water systems supplied by wells. EHP established two sets of flagging criteria in 1986 to assist in the management of sample results. The flagging criteria aid personnel in sample scheduling, interpretation of data, and trend identification. An explanation of flagging criteria for the fourth quarter is presented in the Flagging Criteria sectionmore » of this document. All analytical results from fourth quarter 1989 are listed in this report, which is distributed to all waste-site custodians.« less

The Savannah River Site`s Groundwater Monitoring Program. Fourth quarter, 1989

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

Not Available

1989-12-31

The Environmental Monitoring Section of the Environmental and Health Protection (EHP) Department administers the Savannah River Site`s Groundwater Monitoring Program. During fourth quarter 1989 (October--December), EHP conducted routine sampling of monitoring wells and drinking water locations. EHP collected the drinking water samples from Savannah River Site (SRS) drinking water systems supplied by wells. EHP established two sets of flagging criteria in 1986 to assist in the management of sample results. The flagging criteria aid personnel in sample scheduling, interpretation of data, and trend identification. An explanation of flagging criteria for the fourth quarter is presented in the Flagging Criteria sectionmore » of this document. All analytical results from fourth quarter 1989 are listed in this report, which is distributed to all waste-site custodians.« less

Flagging threshold optimization for manual blood smear review in primary care laboratory.

PubMed

Bihl, Pierre-Adrien

2018-04-01

Manual blood smear review is required when an anomaly detected by the automated hematologic analyzer triggers a flag. Our will through this study is to optimize these flagging thresholds for manual slide review in order to limit workload, while insuring clinical care through no extra false-negative. Flagging causes of 4,373 samples were investigated by manual slide review, after having been run on ADVIA 2120i. A set of 6 user-adjustments is proposed. By implementing all recommendations that we made, false-positive rate falls from 81.8% to 58.6%, while PPV increases from 18.2% to 23.7%. Hence, use of such optimized thresholds enables us to maximize efficiency without altering clinical care, but each laboratory should establish its own criteria to take into consideration local distinctive features.

Construction and characterization of 3A-epitope-tagged foot-and-mouth disease virus.

PubMed

Ma, Xueqing; Li, Pinghua; Sun, Pu; Bai, Xingwen; Bao, Huifang; Lu, Zengjun; Fu, Yuanfang; Cao, Yimei; Li, Dong; Chen, Yingli; Qiao, Zilin; Liu, Zaixin

2015-04-01

Nonstructural protein 3A of foot-and-mouth disease virus (FMDV) is a partially conserved protein of 153 amino acids (aa) in most FMDVs examined to date. Specific deletion in the FMDV 3A protein has been associated with the inability of FMDV to grow in primary bovine cells and cause disease in cattle. However, the aa residues playing key roles in these processes are poorly understood. In this study, we constructed epitope-tagged FMDVs containing an 8 aa FLAG epitope, a 9 aa haemagglutinin (HA) epitope, and a 10 aa c-Myc epitope to substitute residues 94-101, 93-101, and 93-102 of 3A protein, respectively, using a recently developed O/SEA/Mya-98 FMDV infectious cDNA clone. Immunofluorescence assay (IFA), Western blot and sequence analysis showed that the epitope-tagged viruses stably maintained and expressed the foreign epitopes even after 10 serial passages in BHK-21 cells. The epitope-tagged viruses displayed growth properties and plaque phenotypes similar to those of the parental virus in BHK-21 cells. However, the epitope-tagged viruses exhibited lower growth rates and smaller plaque size phenotypes than those of the parental virus in primary fetal bovine kidney (FBK) cells, but similar growth properties and plaque phenotypes to those of the recombinant viruses harboring 93-102 deletion in 3A. These results demonstrate that the decreased ability of FMDV to replicate in primary bovine cells was not associated with the length of 3A, and the genetic determinant thought to play key role in decreased ability to replicate in primary bovine cells could be reduced from 93-102 residues to 8 aa residues at positions 94-101 in 3A protein. Copyright © 2015 Elsevier B.V. All rights reserved.

Targeting Androgen Receptor by Lysosomal Degradation in Prostate Cancer

DTIC Science & Technology

2015-11-01

transport of TFEB, Autophagy 8 (2012) 903-914. [7] J. Brugarolas, K. Lei , R.L. Hurley, B.D. Manning, J.H. Reiling, E. Hafen, L.A. Witters, L.W. Ellisen...chromatography ~ anti-FLAG immunoprecipitation B D EWS pos~t~on mass pept~ de 416-429 1449 .66 GDATVSYEDPPTAK 571-594 2289.08 GGPGGMRGGRGGLMDRGGPGGMFR...vector 1 2 3 -- FLAG-His EWS-Fii-1 4 5 6 - FLAG tubulin c RNA helicase A pos~t~on mass pept~ de 121-141 2161 .97 AENNSEVGASGYGVPGPTWDR 200-209

Analytic Simulation of the Performance of Mobile Maintenance Contact Teams

DTIC Science & Technology

1985-05-01

RECTANGULAR THE CUSTOMERS, REQUIRING MAINT SERVICE, ARE ASSUMED DISTRIBUTED WITHIN A RECTANGLE WITH DIMENSIONS ARANGE ’BY BRANGE . THE POPULATION OF...AND SERVE A CUSTOMER. ’INPUTS: ’FLAG.DIST ’FLAG.FIFO I PR I NT ARANGE ’ BRANGE SPEED 4 POP ’NSERVE MTBF MTTR AN INTEGER FLAG TO INDICATE...TEAMlS). 81 READ ARANGE 32 PRINT ’I LINE THUS INPUT THE DEPTH CCROSS-FRONTAL; DIMENSION (KM; OF THE AREA OF THE TEAMiS;. 84 READ BRANGE

Red eyes and red-flags: improving ophthalmic assessment and referral in primary care

PubMed Central

Kilduff, Caroline; Lois, Charis

2016-01-01

Up to five percent of primary care consultations are eye-related, yet 96% of General Practitioners (GPs) do not undergo postgraduate ophthalmology training. Most do not feel assured performing eye assessments. Some red eye conditions can become sight threatening, and often exhibit red-flag features. These features include moderate pain, photophobia, reduced visual acuity (VA), eye-trauma, or unilateral marked redness. The aim of this project was to improve primary care assessment and referral of patients presenting with red-flag features based on the NICE ‘Red Eye’ Clinical Knowledge Summary recommendations. Data was collected retrospectively from 139 red eye consultations. A practice meeting highlighted poor awareness of red-flag features, low confidence levels in eye assessments, and time-constraints during appointments. Interventions were based on feedback from staff. These included a primary care teaching session on red-flag features, a VA measurement tutorial, and provision of a red eye toolkit, including VA equipment, to each consultation room. At baseline, each patient had on average 0.9 red-flag features assessed. Only 36.0% (9/25) of patients with red-flag features were appropriately referred to same-day ophthalmology services. Following two improvement cycles, a significant improvement was seen in almost every parameter. On average, each patient had 2.7 red-flag features assessed (vs 0.9, p<0.001). VA was assessed in 55.6% of consultations (vs 7.9%, p<0.001), pain was quantified in 81.5% (vs 20.9%, p=0.005), eye-trauma or foreign-body (51.8% vs 8.6%, p<0.001), extent of redness was documented in 66.7% (vs 14.4%, p<0.001). Only photophobia remained poorly assessed (18.5% vs 14.4%, p=0.75). Following this, 75.0% (6/8) of patients were appropriately referred. This project reflected the literature regarding low confidence and inexperience amongst GPs when faced with ophthalmic conditions. Improvements in education are required to ensure accurate assessments can be undertaken in a time-constrained environment. PMID:27493748

Crew Photo in front of flag

NASA Image and Video Library

2010-07-04

ISS024-E-007376 (3 July 2010) --- NASA astronauts Shannon Walker and Doug Wheelock, both Expedition 24 flight engineers, pose for a photo with an American flag while aboard the International Space Station.

APOLLO 17 - FLAG DEDICATION - JSC

NASA Image and Video Library

1974-01-15

S74-15520 --- Left to right Gene Kranz, Gene Cernan, Karla Garnuch, Harrison Schmitt, George Abbey, and Sigurd A. Sjoberg watching the dedication of the Apollo 17 flag to the Mission Control Center. Photo credit: NASA

Interior detail of platform in main hall, with desk, flag, ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Interior detail of platform in main hall, with desk, flag, and banners, facing south - International Longshoremen's & Warehousemen's Union Hall, Naval Civil Engineering Laboratory, Port Hueneme Road, Port Hueneme, Ventura County, CA

Identification of atmospheric fronts over the ocean with microwave measurements of water vapor and rain

NASA Technical Reports Server (NTRS)

Katsaros, Kristina B.; Bhatti, Iftekhar; Mcmurdie, Lynn A.; Patty, Grant W.

1989-01-01

This paper describes some basic research techniques and algorithms developed to diagnose fronts in cyclonic storms over the ocean with data from satellite-borne microwave radiometers. Methods are developed for flagging strong gradients in integrated atmospheric water vapor and the presence of rain by using data from the SSMR on board the polar orbiting Seasat and Nimbus-7 satellites. Examination of 65 frontal systems showed that the water vapor gradient flag correctly identified 86 percent of the fronts, while the precipitation flagged 91 percent. The two types of flags emphasize different portions of the cyclone and are therefore complementary. Ultimately, these techniques are intended for operational use with data from the Special Sensor Microwave Imager which was launched in June 1987 on a satellite in the Defense Meteorological Satellite Program (DMSP).

Water-level database update for the Death Valley regional groundwater flow system, Nevada and California, 1907-2007

USGS Publications Warehouse

Pavelko, Michael T.

2010-01-01

The water-level database for the Death Valley regional groundwater flow system in Nevada and California was updated. The database includes more than 54,000 water levels collected from 1907 to 2007, from more than 1,800 wells. Water levels were assigned a primary flag and multiple secondary flags that describe hydrologic conditions and trends at the time of the measurement and identify pertinent information about the well or water-level measurement. The flags provide a subjective measure of the relative accuracy of the measurements and are used to identify which water levels are appropriate for calculating head observations in a regional transient groundwater flow model. Included in the report appendix are all water-level data and their flags, selected well data, and an interactive spreadsheet for viewing hydrographs and well locations.

Operator-Adjustable Frame Rate, Resolution, and Gray Scale Tradeoff in Fixed-Bandwidth Remote Manipulator Control.

DTIC Science & Technology

1980-09-01

WFLOR(2,3,4) IWAIT FOR FLAG CALL READEF(3,IUU) ICHECK 010 FLAG IF(IUU.NE.2) GO TO 587 IF(.NOT.LCONT) GO TO 999 K-K-54 IMAKE 7,8,9 LOOK LIKE 1,2,3 IF...K.EQ.-6) GO TO 999 IF(K.EQ.3) GO TO 223 IF(K.LT.1) GO TO 223 GO TO 588 587 CALL READEF(4,ITT) ICHECK 5 SEC FLAG IF(ITT.NE.2) GO TO 546 ISKIP IF NOT SET...ISEC,NESC,NYNESC,NESC 1092 FORMAT(’ ’,2A,"-Q’,’TIME 1,12o’s’,I2,2A,’l1, *-86- 1 A, 111A,AW GO TO 222 546 CALL READEF(2,ITS) ICHECK STOP FLAG IF

Vitamin D. Treatment of Prostate Cancer: The Inhibitory Role of IGFBP-3

DTIC Science & Technology

2005-01-01

cassette, sub-family C (CFTR/MRP), member 6 AA424804 Flag 1.62 2.36 Xeroderma pigmentosum , complementation group C (XPC) AA287323 1.44 1.83 2.25...expressed gene I (POVI) T72067 2.37 7.30 Xeroderma pigmentosum , complementation group C (XPC) AA287323 2.25 11.2 Claudin 4 AA506754 2.21 7.30 UDP...Diego: Academic Press; 2001. in DNA damage response ( xeroderma pigmentosum pp 257-303. compledamentatgerouponse C e r [e 3. Miller GJ. Vitamin D and

Overexpression of DJ-1/PARK7, the Parkinson's disease-related protein, improves mitochondrial function via Akt phosphorylation on threonine 308 in dopaminergic neuron-like cells.

PubMed

Zhang, Yi; Gong, Xiao-Gang; Wang, Zhen-Zhen; Sun, Hong-Mei; Guo, Zhen-Yu; Hu, Jing-Hong; Ma, Ling; Li, Ping; Chen, Nai-Hong

2016-05-01

DJ-1/PARK7, the Parkinson's disease-related protein, plays an important role in mitochondrial function. However, the mechanisms by which DJ-1 affects mitochondrial function are not fully understood. Akt is a promoter of neuron survival and is partly involved in the neurodegenerative process. This research aimed at investigating a possible relationship between DJ-1 and Akt signalling in regulating mitochondrial function in the dopaminergic neuron-like cells SH-SY5Y and PC-12. Overexpression of DJ-1 was firstly validated at both the transcriptional and translational levels after transit transfection with plasmid pcDNA3-Flag-DJ-1. Confocal fluorescence microscopy demonstrated that overexpression of DJ-1 increased the mitochondrial mass, but did not disrupt the mitochondrial morphology. In addition, mitochondrial complex I activity was raised in DJ-1-overexpressing cells, and this rise occurred with an increase in cellular adenosine 5'-triphosphate content. Moreover, immunoblotting demonstrated that the levels of phosphoinositide 3-kinase and the total Akt were not altered in DJ-1-overexpressing cells, and nor was the Akt phosphorylation on serine 473 changed. By contrast, Akt phosphorylation on threonine 308 was significantly augmented by overexpression of DJ-1, and the expression of glycogen synthase kinase-3beta, a downstream effector of Akt, was suppressed. In summary, these results suggest that overexpression of DJ-1 improves the mitochondrial function, at least in part, through a mechanism involving Akt phosphorylation on threonine 308. © 2016 Federation of European Neuroscience Societies and John Wiley & Sons Ltd.

Nuclear γ-tubulin associates with nucleoli and interacts with tumor suppressor protein C53.

PubMed

Hořejší, Barbora; Vinopal, Stanislav; Sládková, Vladimíra; Dráberová, Eduarda; Sulimenko, Vadym; Sulimenko, Tetyana; Vosecká, Věra; Philimonenko, Anatoly; Hozák, Pavel; Katsetos, Christos D; Dráber, Pavel

2012-01-01

γ-Tubulin is assumed to be a typical cytosolic protein necessary for nucleation of microtubules from microtubule organizing centers. Using immunolocalization and cell fractionation techniques in combination with siRNAi and expression of FLAG-tagged constructs, we have obtained evidence that γ-tubulin is also present in nucleoli of mammalian interphase cells of diverse cellular origins. Immunoelectron microscopy has revealed γ-tubulin localization outside fibrillar centers where transcription of ribosomal DNA takes place. γ-Tubulin was associated with nucleolar remnants after nuclear envelope breakdown and could be translocated to nucleoli during mitosis. Pretreatment of cells with leptomycin B did not affect the distribution of nuclear γ-tubulin, making it unlikely that rapid active transport via nuclear pores participates in the transport of γ-tubulin into the nucleus. This finding was confirmed by heterokaryon assay and time-lapse imaging of photoconvertible protein Dendra2 tagged to γ-tubulin. Immunoprecipitation from nuclear extracts combined with mass spectrometry revealed an association of γ-tubulin with tumor suppressor protein C53 located at multiple subcellular compartments including nucleoli. The notion of an interaction between γ-tubulin and C53 was corroborated by pull-down and co-immunoprecipitation experiments. Overexpression of γ-tubulin antagonized the inhibitory effect of C53 on DNA damage G(2) /M checkpoint activation. The combined results indicate that aside from its known role in microtubule nucleation, γ-tubulin may also have nuclear-specific function(s). Copyright © 2011 Wiley Periodicals, Inc.

Calcyclin Binding Protein/Siah-1 Interacting Protein Is a Hsp90 Binding Chaperone

PubMed Central

Góral, Agnieszka; Bieganowski, Paweł; Prus, Wiktor; Krzemień-Ojak, Łucja; Kądziołka, Beata; Fabczak, Hanna; Filipek, Anna

2016-01-01

The Hsp90 chaperone activity is tightly regulated by interaction with many co-chaperones. Since CacyBP/SIP shares some sequence homology with a known Hsp90 co-chaperone, Sgt1, in this work we performed a set of experiments in order to verify whether CacyBP/SIP can interact with Hsp90. By applying the immunoprecipitation assay we have found that CacyBP/SIP binds to Hsp90 and that the middle (M) domain of Hsp90 is responsible for this binding. Furthermore, the proximity ligation assay (PLA) performed on HEp-2 cells has shown that the CacyBP/SIP-Hsp90 complexes are mainly localized in the cytoplasm of these cells. Using purified proteins and applying an ELISA we have shown that Hsp90 interacts directly with CacyBP/SIP and that the latter protein does not compete with Sgt1 for the binding to Hsp90. Moreover, inhibitors of Hsp90 do not perturb CacyBP/SIP-Hsp90 binding. Luciferase renaturation assay and citrate synthase aggregation assay with the use of recombinant proteins have revealed that CacyBP/SIP exhibits chaperone properties. Also, CacyBP/SIP-3xFLAG expression in HEp-2 cells results in the appearance of more basic Hsp90 forms in 2D electrophoresis, which may indicate that CacyBP/SIP dephosphorylates Hsp90. Altogether, the obtained results suggest that CacyBP/SIP is involved in regulation of the Hsp90 chaperone machinery. PMID:27249023

Exposure to the American flag polarizes democratic-republican ideologies.

PubMed

Chan, Eugene Y

2017-12-01

Some prior research has suggested that exposure to the American flag tilts Americans towards Republicanism, while others have proffered that it brings outs a common 'together' perspective instead. We explore a third possibility - that it may actually polarize Americans' political ideology. It is generally accepted that exposure to an environmental cue can shift attitudes and behaviours, at least partly or temporarily, in a manner that is consistent with that cue. Yet, the same cue can mean different things to different people. In the same vein, given how national identity and political ideology are intertwined in the United States, we hypothesize that the American flag should heighten different political beliefs depending on individuals' political ideology. To Democrats, being American is to support Democratic values, but to Republicans, being American is to support Republican values. The American flag thus should heighten Democrats of their Democratic identity, and it should heighten Republicans of their Republican one. The results of an experiment with 752 American respondents who were representative of the US population supported this polarizing effect of the American flag. The theoretical and policy implications of the findings are offered. © 2017 The British Psychological Society.

Ground squirrel tail-flag displays alter both predatory strike and ambush site selection behaviours of rattlesnakes.

PubMed

Barbour, Matthew A; Clark, Rulon W

2012-09-22

Many species approach, inspect and signal towards their predators. These behaviours are often interpreted as predator-deterrent signals--honest signals that indicate to a predator that continued hunting is likely to be futile. However, many of these putative predator-deterrent signals are given when no predator is present, and it remains unclear if and why such signals deter predators. We examined the effects of one such signal, the tail-flag display of California ground squirrels, which is frequently given both during and outside direct encounters with northern Pacific rattlesnakes. We video-recorded and quantified the ambush foraging responses of rattlesnakes to tail-flagging displays from ground squirrels. We found that tail-flagging deterred snakes from striking squirrels, most likely by advertising squirrel vigilance (i.e. readiness to dodge a snake strike). We also found that tail-flagging by adult squirrels increased the likelihood that snakes would leave their ambush site, apparently by elevating the vigilance of nearby squirrels which reduces the profitability of the ambush site. Our results provide some of the first empirical evidence of the mechanisms by which a prey display, although frequently given in the absence of a predator, may still deter predators during encounters.

COOPER, HERON, AND HEWARD'S APPLIED BEHAVIOR ANALYSIS (2ND ED.): CHECKERED FLAG FOR STUDENTS AND PROFESSORS, YELLOW FLAG FOR THE FIELD

PubMed Central

Friman, Patrick C

2010-01-01

At last, the field of applied behavior analysis has a beautifully crafted, true textbook that can proudly stand cover to cover and spine to spine beside any of the expensive, imposing, and ornately designed textbooks used by college instructors who teach courses in conventional areas of education or psychology. In this review, I fully laud this development, credit Cooper, Heron, and Heward for making it happen, argue that it signifies a checkered flag for students and professors, and recommend the book for classes in applied behavior analysis everywhere. Subsequently, I review its chapters, each of which could easily stand alone as publications in their own right. Finally, I supply a cautionary note, a yellow flag to accompany the well-earned checkered flag, by pointing out that, as is true with all general textbooks on applied behavior analysis, a major portion of the references involves research on persons who occupy only a tail of the normal distribution. To attain the mainstream role Skinner envisioned and most (if not all) behavior analysts desire, the field will have to increase its focus on persons who reside under the dome of that distribution.

Cloud based, Open Source Software Application for Mitigating Herbicide Drift

NASA Astrophysics Data System (ADS)

Saraswat, D.; Scott, B.

2014-12-01

The spread of herbicide resistant weeds has resulted in the need for clearly marked fields. In response to this need, the University of Arkansas Cooperative Extension Service launched a program named Flag the Technology in 2011. This program uses color-coded flags as a visual alert of the herbicide trait technology within a farm field. The flag based program also serves to help avoid herbicide misapplication and prevent herbicide drift damage between fields with differing crop technologies. This program has been endorsed by Southern Weed Science Society of America and is attracting interest from across the USA, Canada, and Australia. However, flags have risk of misplacement or disappearance due to mischief or severe windstorms/thunderstorms, respectively. This presentation will discuss the design and development of a cloud-based, free application utilizing open-source technologies, called Flag the Technology Cloud (FTTCloud), for allowing agricultural stakeholders to color code their farm fields for indicating herbicide resistant technologies. The developed software utilizes modern web development practices, widely used design technologies, and basic geographic information system (GIS) based interactive interfaces for representing, color-coding, searching, and visualizing fields. This program has also been made compatible for a wider usability on different size devices- smartphones, tablets, desktops and laptops.

STS-51D - CREW INSIGNIA

NASA Image and Video Library

1985-01-01

S85-28989 (March 1985) --- The dominant features of the STS-51D emblem are an orbit formed by a Colonial American flag and a space shuttle. The flag in orbit signifies the U.S. flag to indicate that it comes from this country and the American people. The original 13-star flag is used to symbolize a continuity of technical achievement and progress since colonial times. The name Discovery preceding the flag represents the spirit of Discovery and exploration of new frontiers which have been a hallmark of American people even before they were formed together as a nation. The crew members are Karol J. Bobko, Donald E. Williams, Rhea Seddon, S. David Griggs and Jeffrey A. Hoffman of NASA; and Charles D. Walker, representing McDonnell Douglas Corporation; and U. S. Senator Jake Garn. The NASA insignia design for space shuttle flights is reserved for use by the astronauts and for other official use as the NASA Administrator may authorize. Public availability has been approved only in the forms of illustrations by the various news media. When and if there is any change in this policy, which is not anticipated, the change will be publicly announced. Photo credit: NASA

38. FLAG/ADMIRAL BRIDGE PORT LOOKING TO STARBOARD SHOWING RADAR ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

38. FLAG/ADMIRAL BRIDGE - PORT LOOKING TO STARBOARD SHOWING RADAR SCOPE, ADMIRAL'S CHAIR, GYRO REPEATER AND VARIOUS COMMUNICATION SYSTEMS. - U.S.S. HORNET, Puget Sound Naval Shipyard, Sinclair Inlet, Bremerton, Kitsap County, WA

6. ADMINISTRATION BUILDING WITH FLAG POLE, LOOKING SOUTH. NIKE ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

6. ADMINISTRATION BUILDING WITH FLAG POLE, LOOKING SOUTH. - NIKE Missile Base SL-40, Administration Building, East central portion of base, southeast of Mess Hall, northeast of HIPAR Equipment Building, Hecker, Monroe County, IL

(U) A Gruneisen Equation of State for TPX. Application in FLAG

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

Fredenburg, David A.; Aslam, Tariq Dennis; Bennett, Langdon Stanford

2015-11-02

A Gruneisen equation of state (EOS) is developed for the polymer TPX (poly 4-methyl-1-pentene) within the LANL hydrocode FLAG. Experimental shock Hugoniot data for TPX is fit to a form of the Gruneisen EOS, and the necessary parameters for implementing the TPX EOS in FLAG are presented. The TPX EOS is further validated through one-dimensional simulations of recent double-shock experiments, and a comparison is made between the new Gruneisen EOS for TPX and the EOS representation for TPX used in the LANL Common Model.

KSC-07pd0766

NASA Image and Video Library

2007-04-02

KENNEDY SPACE CENTER, FLA. -- The American flag and the NASA logo shine in the morning sun on the side of the Vehicle Assembly Building after completion of their repainting. The flag spans an area 209 feet by 110 feet, or about 23, 437 square feet. Each stripe is 9 feet wide and each star is 6 feet in diameter. The NASA logo, which is known as the "meatball," measures 110 feet by 132 feet, or about 12,300 square feet. The flag and logo were last painted in 1998, honoring NASA's 40th anniversary. Photo credit: NASA/Jim Grossmann

Pedagog II Realization.

DTIC Science & Technology

1979-12-01

BE2 CD66 ALl SGG/110(14) 5/AS2 SGG/79(14) AMI 110(13) 5/AU2 78(16) NI KRIRF/lll(2) ANl PRIRF/lll(l) ARI IPC/lll(ll) 7/BPl IPC/144(4) ASI KRF/106(9...KRS 6034 Read a character from the keyboard/reader buffer. The keyboard/reader flag is set when the operation is completed. KIE 6035 Enable the...keyboard/ reader flag is a 1. KCC 6032 Clear the AC and the keyboard/reader flag. KRS 6034 Read a character from the keyboard/reader buffer. The keyboard

Comparison of five automated hematology analyzers in a university hospital setting: Abbott Cell-Dyn Sapphire, Beckman Coulter DxH 800, Siemens Advia 2120i, Sysmex XE-5000, and Sysmex XN-2000.

PubMed

Bruegel, Mathias; Nagel, Dorothea; Funk, Manuela; Fuhrmann, Petra; Zander, Johannes; Teupser, Daniel

2015-06-01

Various types of automated hematology analyzers are used in clinical laboratories. Here, we performed a side-by-side comparison of five current top of the range routine hematology analyzers in the setting of a university hospital central laboratory. Complete blood counts (CBC), differentials, reticulocyte and nucleated red blood cell (NRBC) counts of 349 patient samples, randomly taken out of routine diagnostics, were analyzed with Cell-Dyn Sapphire (Abbott), DxH 800 (Beckman Coulter), Advia 2120i (Siemens), XE-5000 and XN-2000 (Sysmex). Inter-instrument comparison of CBCs including reticulocyte and NRBC counts and investigation of flagging quality in relation to microscopy were performed with the complete set of samples. Inter-instrument comparison of five-part differential was performed using samples without atypical cells in blood smear (n=292). Automated five-part differentials and NRBCs were additionally compared with microscopy. The five analyzers showed a good concordance for basic blood count parameters. Correlations between instruments were less well for reticulocyte counts, NRBCs, and differentials. The poorest concordance for NRBCs with microscopy was observed for Advia 2120i (Kendall's τb=0.37). The highest flagging sensitivity for blasts was observed for XN-2000 (97% compared to 65%-76% for other analyzers), whereas overall specificity was comparable between different instruments. To the best of our knowledge, this is the most comprehensive side-by-side comparison of five current top of the range routine hematology analyzers. Variable analyzer quality and parameter specific limitations must be considered in defining laboratory algorithms in clinical practice.

Gasdermin D is an executor of pyroptosis and required for interleukin-1β secretion

PubMed Central

He, Wan-ting; Wan, Haoqiang; Hu, Lichen; Chen, Pengda; Wang, Xin; Huang, Zhe; Yang, Zhang-Hua; Zhong, Chuan-Qi; Han, Jiahuai

2015-01-01

Inflammasome is an intracellular signaling complex of the innate immune system. Activation of inflammasomes promotes the secretion of interleukin 1β (IL-1β) and IL-18 and triggers pyroptosis. Caspase-1 and -11 (or -4/5 in human) in the canonical and non-canonical inflammasome pathways, respectively, are crucial for inflammasome-mediated inflammatory responses. Here we report that gasdermin D (GSDMD) is another crucial component of inflammasomes. We discovered the presence of GSDMD protein in nigericin-induced NLRP3 inflammasomes by a quantitative mass spectrometry-based analysis. Gene deletion of GSDMD demonstrated that GSDMD is required for pyroptosis and for the secretion but not proteolytic maturation of IL-1β in both canonical and non-canonical inflammasome responses. It was known that GSDMD is a substrate of caspase-1 and we showed its cleavage at the predicted site during inflammasome activation and that this cleavage was required for pyroptosis and IL-1β secretion. Expression of the N-terminal proteolytic fragment of GSDMD can trigger cell death and N-terminal modification such as tagging with Flag sequence disrupted the function of GSDMD. We also found that pro-caspase-1 is capable of processing GSDMD and ASC is not essential for GSDMD to function. Further analyses of LPS plus nigericin- or Salmonella typhimurium-treated macrophage cell lines and primary cells showed that apoptosis became apparent in Gsdmd−/− cells, indicating a suppression of apoptosis by pyroptosis. The induction of apoptosis required NLRP3 or other inflammasome receptors and ASC, and caspase-1 may partially contribute to the activation of apoptotic caspases in Gsdmd−/− cells. These data provide new insights into the molecular mechanisms of pyroptosis and reveal an unexpected interplay between apoptosis and pyroptosis. PMID:26611636

Gasdermin D is an executor of pyroptosis and required for interleukin-1β secretion.

PubMed

He, Wan-ting; Wan, Haoqiang; Hu, Lichen; Chen, Pengda; Wang, Xin; Huang, Zhe; Yang, Zhang-Hua; Zhong, Chuan-Qi; Han, Jiahuai

2015-12-01

Inflammasome is an intracellular signaling complex of the innate immune system. Activation of inflammasomes promotes the secretion of interleukin 1β (IL-1β) and IL-18 and triggers pyroptosis. Caspase-1 and -11 (or -4/5 in human) in the canonical and non-canonical inflammasome pathways, respectively, are crucial for inflammasome-mediated inflammatory responses. Here we report that gasdermin D (GSDMD) is another crucial component of inflammasomes. We discovered the presence of GSDMD protein in nigericin-induced NLRP3 inflammasomes by a quantitative mass spectrometry-based analysis. Gene deletion of GSDMD demonstrated that GSDMD is required for pyroptosis and for the secretion but not proteolytic maturation of IL-1β in both canonical and non-canonical inflammasome responses. It was known that GSDMD is a substrate of caspase-1 and we showed its cleavage at the predicted site during inflammasome activation and that this cleavage was required for pyroptosis and IL-1β secretion. Expression of the N-terminal proteolytic fragment of GSDMD can trigger cell death and N-terminal modification such as tagging with Flag sequence disrupted the function of GSDMD. We also found that pro-caspase-1 is capable of processing GSDMD and ASC is not essential for GSDMD to function. Further analyses of LPS plus nigericin- or Salmonella typhimurium-treated macrophage cell lines and primary cells showed that apoptosis became apparent in Gsdmd(-/-) cells, indicating a suppression of apoptosis by pyroptosis. The induction of apoptosis required NLRP3 or other inflammasome receptors and ASC, and caspase-1 may partially contribute to the activation of apoptotic caspases in Gsdmd(-/-) cells. These data provide new insights into the molecular mechanisms of pyroptosis and reveal an unexpected interplay between apoptosis and pyroptosis.

Influence of deer abundance on the abundance of questing adult Ixodes scapularis (Acari: Ixodidae)

USGS Publications Warehouse

Ginsberg, H.S.; Zhioua, E.

1999-01-01

Nymphal and adult Ixodes scapularis Say were sampled by flagging at 2 sites on a barrier island, Fire Island, NY, and at 2 sites on the nearby mainland. Nymphal densities did not differ consistently between island and mainland sites, but adult densities were consistently lower on the island. We tested whether lower adult densities on the island resulted from greater nymphal mortality on the island than the mainland, or whether adult ticks on the island were poorly sampled by flagging because they had attached abundantly to deer, which were common on Fire Island. Differential nymphal mortality on islands vs. mainland did not explain this difference in adult densities because survival of flat and engorged nymphs in enclosures was the same at island and mainland sites. Ticks were infected by parasitic wasps on the island and not the mainland, but the infection rate (4.3%) was too low to explain the difference in adult tick densities. In contrast, exclusion of deer by game fencing on Fire Island resulted in markedly increased numbers of adult ticks in flagging samples inside compared to samples taken outside the exclosures. Therefore, the scarcity of adult ticks in flagging samples on Fire Island resulted, at least in part, from the ticks being unavailable to flagging samples because they were on deer hosts. Differences in the densities of flagged ticks inside and outside the exclosures were used to estimate the percentage of questing adults on Fire Island that found deer hosts, excluding those that attached to other host species. Approximately 56% of these questing adult ticks found deer hosts in 1995 and 50% found deer hosts in 1996. Therefore, in areas where vertebrate hosts are highly abundant, large proportions of the questing tick population can find hosts. Moreover, comparisons of tick densities at different sites by flagging can potentially be biased by differences in host densities among sites.

Cost and effects of different admission screening strategies to control the spread of methicillin-resistant Staphylococcus aureus.

PubMed

Gurieva, Tanya; Bootsma, Martin C J; Bonten, Marc J M

2013-01-01

Nosocomial infection rates due to antibiotic-resistant bacteriae, e.g., methicillin-resistant Staphylococcus aureus (MRSA) remain high in most countries. Screening for MRSA carriage followed by barrier precautions for documented carriers (so-called screen and isolate (S&I)) has been successful in some, but not all settings. Moreover, different strategies have been proposed, but comparative studies determining their relative effects and costs are not available. We, therefore, used a mathematical model to evaluate the effect and costs of different S&I strategies and to identify the critical parameters for this outcome. The dynamic stochastic simulation model consists of 3 hospitals with general wards and intensive care units (ICUs) and incorporates readmission of carriers of MRSA. Patient flow between ICUs and wards was based on real observations. Baseline prevalence of MRSA was set at 20% in ICUs and hospital-wide at 5%; ranges of costs and infection rates were based on published data. Four S&I strategies were compared to a do-nothing scenario: S&I of previously documented carriers ("flagged" patients); S&I of flagged patients and ICU admissions; S&I of flagged and group of "frequent" patients; S&I of all hospital admissions (universal screening). Evaluated levels of efficacy of S&I were 10%, 25%, 50% and 100%. Our model predicts that S&I of flagged and S&I of flagged and ICU patients are the most cost-saving strategies with fastest return of investment. For low isolation efficacy universal screening and S&I of flagged and "frequent" patients may never become cost-saving. Universal screening is predicted to prevent hardly more infections than S&I of flagged and "frequent" patients, albeit at higher costs. Whether an intervention becomes cost-saving within 10 years critically depends on costs per infection in ICU, costs of screening and isolation efficacy.

Expression, purification, and functional characterization of the insulin-responsive facilitative glucose transporter GLUT4.

PubMed

Kraft, Thomas E; Hresko, Richard C; Hruz, Paul W

2015-12-01

The insulin-responsive facilitative glucose transporter GLUT4 is of fundamental importance for maintenance of glucose homeostasis. Despite intensive effort, the ability to express and purify sufficient quantities of structurally and functionally intact protein for biophysical analysis has previously been exceedingly difficult. We report here the development of novel methods to express, purify, and functionally reconstitute GLUT4 into detergent micelles and proteoliposomes. Rat GLUT4 containing FLAG and His tags at the amino and carboxy termini, respectively, was engineered and stably transfected into HEK-293 cells. Overexpression in suspension culture yielded over 1.5 mg of protein per liter of culture. Systematic screening of detergent solubilized GLUT4-GFP fusion protein via fluorescent-detection size exclusion chromatography identified lauryl maltose neopentyl glycol (LMNG) as highly effective for isolating monomeric GLUT4 micelles. Preservation of structural integrity and ligand binding was demonstrated via quenching of tryptophan fluorescence and competition of ATB-BMPA photolabeling by cytochalasin B. GLUT4 was reconstituted into lipid nanodiscs and proper folding was confirmed. Reconstitution of purified GLUT4 with amphipol A8-35 stabilized the transporter at elevated temperatures for extended periods of time. Functional activity of purified GLUT4 was confirmed by reconstitution of LMNG-purified GLUT4 into proteoliposomes and measurement of saturable uptake of D-glucose over L-glucose. Taken together, these data validate the development of an efficient means to generate milligram quantities of stable and functionally intact GLUT4 that is suitable for a wide array of biochemical and biophysical analyses. © 2015 The Protein Society.

Pseudo-Kähler Quantization on Flag Manifolds

NASA Astrophysics Data System (ADS)

Karabegov, Alexander V.

A unified approach to geometric, symbol and deformation quantizations on a generalized flag manifold endowed with an invariant pseudo-Kähler structure is proposed. In particular cases we arrive at Berezin's quantization via covariant and contravariant symbols.

Bernard J. Cigrand, DDS: Father of Flag Day and renaissance man.

PubMed

Spiegel, Allen D; Kavaler, Florence

2007-06-01

Dr. Bernard J. Cigrand is acknowledged as the "Father of Flag Day." He relentlessly continued his activities for more than sixty years to have June 14 designated for the national observance of the birth of the American flag. That finally occurred in 1948, seventeen years after his death, when President Harry S. Truman signed a Congressional Act into law. However, the law designated a voluntary observance but did not create a legal national holiday. In addition to his fervent passion for Flag Day, Cigrand undertook a variety of other initiatives He was a practicing dentist, the dean of a dental school, an investigative journalist, an expert on heraldry and seals, a lecturer and an author of books. Despite all his achievements, Cigrand is more widely known in the European country from where his parents emigrated to the United States in 1852 - Luxembourg, than in the United States.

KSC-98pc903

NASA Image and Video Library

1998-08-11

Painters are suspended on platforms from the top of the 525-foot-high Vehicle Assembly Building (VAB) at KSC during repainting of the American flag. The flag spans an area 209 feet by 110 feet and will require 510 gallons of red, white and blue paint. Each stripe of the flag is 9 feet wide and each star is 6 feet in diameter. The platforms are operated by two electric motors and travel 35 feet per minute. Work is being done with rollers, with brushes being used for details. The paint was donated by ICI Devoe of Louisville, Ky. In addition to the flag, the Bicentennial Emblem on the other side of the VAB doors is being replaced by the NASA logo, honoring NASA’s 40th anniversary (in October). The logo covers an area 110 feet by 132 feet. Work is expected to be completed in mid-September

The American flag on the VAB is being repainted

NASA Technical Reports Server (NTRS)

1998-01-01

Painters are suspended on platforms from the top of the 525-foot- high Vehicle Assembly Building (VAB) at KSC during repainting of the American flag and NASA logo. The flag spans an area 209 feet by 110 feet and will require 510 gallons of red, white and blue paint. Each stripe of the flag is 9 feet wide and each star is 6 feet in diameter. The previous Bicentennial Emblem on the other side of the VAB doors is being replaced by the NASA logo, honoring NASA's 40th anniversary (in October). The logo covers an area 110 feet by 132 feet. The painting platforms are operated by two electric motors and travel 35 feet per minute. Work is being done with rollers, with brushes being used for details. The paint was donated by ICI Devoe of Louisville, Ky. Work is expected to be completed in mid-September.

The influence of inorganic nitrogen fertilizer forms on micronutrient retranslocation and accumulation in grains of winter wheat.

PubMed

Barunawati, Nunun; Giehl, Ricardo F Hettwer; Bauer, Bernhard; von Wirén, Nicolaus

2013-01-01

The fortification of cereal grains with metal micronutrients is a major target to combat human malnutrition of Fe and Zn. Based on recent studies showing that N fertilization can promote Fe and Zn accumulation in cereal grains, we investigated here the influence of nitrate- or ammonium-based N fertilization on the accumulation of Fe, Zn, and Cu as well as metal chelator pools in flag leaves and grains of winter wheat. Fertilization with either N form increased the concentrations of N and of the metal chelator nicotianamine (NA) in green leaves, while 2'-deoxymugineic acid (DMA) remained unaffected. Despite the differential response to N fertilization of NA and DMA levels in flag leaves, N fertilization remained without any significant effect on the net export of these metals during flag leaf senescence, which accounted for approximately one third of the total Fe, Zn, or Cu content in leaves. The significant increase in the accumulation of Fe, Zn, and Cu found in the grains of primarily ammonium-fertilized plants was unrelated to the extent of metal retranslocation from flag leaves. These results indicate that an increased N nutritional status of flag leaves promotes the accumulation of Fe, Zn, and Cu in flag leaves, which is accompanied by an increased pool of NA but not of DMA. With regard to the far higher concentrations of DMA relative to NA in leaves and leaf exudates, DMA may be more relevant for the mobilization and retranslocation of these metals in high-yielding wheat production.

The impact of testing accommodations on MCAT scores: descriptive results.

PubMed

Julian, Ellen R; Ingersoll, Deborah J; Etienne, Patricia M; Hilger, Anthony E

2004-04-01

Medical College Admission Test (MCAT) examinees with disabilities who receive accommodations receive flagged scores indicating nonstandard administration. This report compares MCAT examinees who received accommodations and their performances with standard examinees. Aggregate history records of all 1994-2000 MCAT examinees were identified as flagged (2,401) or standard (297,880), then further sorted by race/ethnicity (broadly identified as underrepresented minority and non-URM, at the time of testing) and gender. Those with flagged scores were also classified by disability (LD = learning disability, ADHD = attention deficit hyperactivity disorder, LD/ADHD = learning disability and attention deficit hyperactivity disorder, and Other = other disability) and type of accommodation. Mean MCAT scores were calculated for all groups. A group of 866 examinees took the MCAT first as a standard administration and subsequently with accommodations. In a separate analysis, their two sets of scores were compared. Less than 1% of examinees (2,401) had accommodations; of these, 55% were LD, 17% ADHD, 5% LD/ADHD, and 23% Other. Extended time was the most frequently provided accommodation. Mean flagged scores slightly exceeded mean standard scores on all MCAT sections. Examinees who retook the MCAT with accommodations after a standard administration increased their scores by six points, quadrupling the average gain Standard-Standard retest cohort from another study. The small but statistically significant different higher flagged scores may reflect either appropriate compensation or overly generous accommodations. Extended time had a positive impact on the scores of those who retested with this accommodation. The validity the flagged MCAT in predicting success in medical school is not known, and further investigation is underway.

[Efficient genome editing in human pluripotent stem cells through CRISPR/Cas9].

PubMed

Liu, Gai-gai; Li, Shuang; Wei, Yu-da; Zhang, Yong-xian; Ding, Qiu-rong

2015-11-01

The RNA-guided CRISPR (clustered regularly interspaced short palindromic repeat)-associated Cas9 nuclease has offered a new platform for genome editing with high efficiency. Here, we report the use of CRISPR/Cas9 technology to target a specific genomic region in human pluripotent stem cells. We show that CRISPR/Cas9 can be used to disrupt a gene by introducing frameshift mutations to gene coding region; to knock in specific sequences (e.g. FLAG tag DNA sequence) to targeted genomic locus via homology directed repair; to induce large genomic deletion through dual-guide multiplex. Our results demonstrate the versatile application of CRISPR/Cas9 in stem cell genome editing, which can be widely utilized for functional studies of genes or genome loci in human pluripotent stem cells.

Squirrel Damage to Pines

Treesearch

USDA Forest Service

1981-01-01

Flagging (dead branch tips) on jack pine and red pine may be caused by insects, diseases, or mechanical damage. In the Lake States, flagging is often the result of mechanical damage, sometimes girdling, caused when the cones are torn off by red squirrels.

78 FR 35101 - Flag Day and National Flag Week, 2013

Federal Register 2010, 2011, 2012, 2013, 2014

2013-06-12

... Glory has followed, reminding us of the rights and responsibilities we share as citizens. This week, we celebrate that legacy, and we honor the brave men and women who have secured it through centuries of service...

46 CFR 154.17 - U.S. flag vessel: Certificate of Inspection endorsement.

Code of Federal Regulations, 2013 CFR

2013-10-01

... DANGEROUS CARGOES SAFETY STANDARDS FOR SELF-PROPELLED VESSELS CARRYING BULK LIQUEFIED GASES General § 154.17.... flag vessel allowed to carry a liquefied gas listed in Table 4 has the following endorsement for each...

46 CFR 154.17 - U.S. flag vessel: Certificate of Inspection endorsement.

Code of Federal Regulations, 2014 CFR

2014-10-01

... DANGEROUS CARGOES SAFETY STANDARDS FOR SELF-PROPELLED VESSELS CARRYING BULK LIQUEFIED GASES General § 154.17.... flag vessel allowed to carry a liquefied gas listed in Table 4 has the following endorsement for each...

46 CFR 154.17 - U.S. flag vessel: Certificate of Inspection endorsement.

Code of Federal Regulations, 2012 CFR

2012-10-01

... DANGEROUS CARGOES SAFETY STANDARDS FOR SELF-PROPELLED VESSELS CARRYING BULK LIQUEFIED GASES General § 154.17.... flag vessel allowed to carry a liquefied gas listed in Table 4 has the following endorsement for each...

Performance evaluation of the new hematology analyzer Sysmex XN-series.

PubMed

Seo, J Y; Lee, S-T; Kim, S-H

2015-04-01

The Sysmex XN-series is a new automated hematology analyzer designed to improve the accuracy of cell counts and the specificity of the flagging events. The basic characteristics and the performance of new measurement channels of the XN were evaluated and compared with the Sysmex XE-2100 and the manual method. Fluorescent platelet count (PLT-F) was compared with the flow cytometric method. The low WBC mode and body fluid mode were also evaluated. For workflow analysis, 1005 samples were analyzed on both the XN and the XE-2100, and manual review rates were compared. All parameters measured by the XN correlated well with the XE-2100. PLT-F showed better correlation with the flow cytometric method (r(2)  = 0.80) compared with optical platelet count (r(2)  = 0.73) for platelet counts <70 × 10(9) /L. The low WBC mode reported accurate leukocyte differentials for samples with a WBC count <0.5 × 10(9) /L. Relatively good correlation was found for WBC counts between the manual method and the body fluid mode (r = 0.88). The XN made less flags than the XE-2100, while the sensitivities of both instruments were comparable. The XN provided reliable results on low cell counts, as well as reduced manual blood film reviews, while maintaining a proper level of diagnostic sensitivity. © 2014 John Wiley & Sons Ltd.

A multipurpose fusion tag derived from an unstructured and hyperacidic region of the amyloid precursor protein

PubMed Central

Sangawa, Takeshi; Tabata, Sanae; Suzuki, Kei; Saheki, Yasushi; Tanaka, Keiji; Takagi, Junichi

2013-01-01

Expression and purification of aggregation-prone and disulfide-containing proteins in Escherichia coli remains as a major hurdle for structural and functional analyses of high-value target proteins. Here, we present a novel gene-fusion strategy that greatly simplifies purification and refolding procedure at very low cost using a unique hyperacidic module derived from the human amyloid precursor protein. Fusion with this polypeptide (dubbed FATT for Flag-Acidic-Target Tag) results in near-complete soluble expression of variety of extracellular proteins, which can be directly refolded in the crude bacterial lysate and purified in one-step by anion exchange chromatography. Application of this system enabled preparation of functionally active extracellular enzymes and antibody fragments without the need for condition optimization. PMID:23526492

Purification of recombinant budgerigar fledgling disease virus VP1 capsid protein and its ability for in vitro capsid assembly

NASA Technical Reports Server (NTRS)

Rodgers, R. E.; Chang, D.; Cai, X.; Consigli, R. A.; Spooner, B. S. (Principal Investigator)

1994-01-01

A recombinant system for the major capsid VP1 protein of budgerigar fledgling disease virus has been established. The VP1 gene was inserted into a truncated form of the pFlag-1 vector and expressed in Escherichia coli. The budgerigar fledgling disease virus VP1 protein was purified to near homogeneity by immunoaffinity chromatography. Fractions containing highly purified VP1 were pooled and found to constitute 3.3% of the original E. coli-expressed VP1 protein. Electron microscopy revealed that the VP1 protein was isolated as pentameric capsomeres. Electron microscopy also revealed that capsid-like particles were formed in vitro from purified VP1 capsomeres with the addition of Ca2+ ions and the removal of chelating and reducing agents.

Effect of the down-regulation of the high Grain Protein Content (GPC) genes on the wheat transcriptome during monocarpic senescence.

PubMed

Cantu, Dario; Pearce, Stephen P; Distelfeld, Assaf; Christiansen, Michael W; Uauy, Cristobal; Akhunov, Eduard; Fahima, Tzion; Dubcovsky, Jorge

2011-10-07

Increasing the nutrient concentration of wheat grains is important to ameliorate nutritional deficiencies in many parts of the world. Proteins and nutrients in the wheat grain are largely derived from the remobilization of degraded leaf molecules during monocarpic senescence. The down-regulation of the NAC transcription factor Grain Protein Content (GPC) in transgenic wheat plants delays senescence (>3 weeks) and reduces the concentration of protein, Zn and Fe in the grain (>30%), linking senescence and nutrient remobilization.Based on the early and rapid up-regulation of GPC in wheat flag leaves after anthesis, we hypothesized that this transcription factor is an early regulator of monocarpic senescence. To test this hypothesis, we used high-throughput mRNA-seq technologies to characterize the effect of the GPC down-regulation on the wheat flag-leaf transcriptome 12 days after anthesis. At this early stage of senescence GPC transcript levels are significantly lower in transgenic GPC-RNAi plants than in the wild type, but there are still no visible phenotypic differences between genotypes. We generated 1.4 million 454 reads from early senescing flag leaves (average ~350 nt) and assembled 1.2 million into 30,497 contigs that were used as a reference to map 145 million Illumina reads from three wild type and four GPC-RNAi plants. Following normalization and statistical testing, we identified a set of 691 genes differentially regulated by GPC (431 ≥ 2-fold change). Transcript level ratios between transgenic and wild type plants showed a high correlation (R = 0.83) between qRT-PCR and Illumina results, providing independent validation of the mRNA-seq approach. A set of differentially expressed genes were analyzed across an early senescence time-course. Monocarpic senescence is an active process characterized by large-scale changes in gene expression which begins considerably before the appearance of visual symptoms of senescence. The mRNA-seq approach used here was able to detect small differences in transcript levels during the early stages of senescence. This resulted in an extensive list of GPC-regulated genes, which includes transporters, hormone regulated genes, and transcription factors. These GPC-regulated genes, particularly those up-regulated during senescence, provide valuable entry points to dissect the early stages of monocarpic senescence and nutrient remobilization in wheat.

High-throughput spectral and lifetime-based FRET screening in living cells to identify small-molecule effectors of SERCA

PubMed Central

Schaaf, Tory M.; Peterson, Kurt C.; Grant, Benjamin D.; Bawaskar, Prachi; Yuen, Samantha; Li, Ji; Muretta, Joseph M.; Gillispie, Gregory D.; Thomas, David D.

2017-01-01

A robust high-throughput screening (HTS) strategy has been developed to discover small-molecule effectors targeting the sarco/endoplasmic reticulum calcium ATPase (SERCA), based on a fluorescence microplate reader that records both the nanosecond decay waveform (lifetime mode) and the complete emission spectrum (spectral mode), with high precision and speed. This spectral unmixing plate reader (SUPR) was used to screen libraries of small molecules with a fluorescence resonance energy transfer (FRET) biosensor expressed in living cells. Ligand binding was detected by FRET associated with structural rearrangements of green (GFP, donor) and red (RFP, acceptor) fluorescent proteins fused to the cardiac-specific SERCA2a isoform. The results demonstrate accurate quantitation of FRET along with high precision of hit identification. Fluorescence lifetime analysis resolved SERCA’s distinct structural states, providing a method to classify small-molecule chemotypes on the basis of their structural effect on the target. The spectral analysis was also applied to flag interference by fluorescent compounds. FRET hits were further evaluated for functional effects on SERCA’s ATPase activity via both a coupled-enzyme assay and a FRET-based calcium sensor. Concentration-response curves indicated excellent correlation between FRET and function. These complementary spectral and lifetime FRET detection methods offer an attractive combination of precision, speed, and resolution for HTS. PMID:27899691

Structural basis underlying CAC RNA recognition by the RRM domain of dimeric RNA-binding protein RBPMS

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

Teplova, Marianna; Farazi, Thalia A.; Tuschl, Thomas

Abstract RNA-binding protein with multiple splicing (designated RBPMS) is a higher vertebrate mRNA-binding protein containing a single RNA recognition motif (RRM). RBPMS has been shown to be involved in mRNA transport, localization and stability, with key roles in axon guidance, smooth muscle plasticity, as well as regulation of cancer cell proliferation and migration. We report on structure-function studies of the RRM domain of RBPMS bound to a CAC-containing single-stranded RNA. These results provide insights into potential topologies of complexes formed by the RBPMS RRM domain and the tandem CAC repeat binding sites as detected by photoactivatable-ribonucleoside-enhanced crosslinking and immunoprecipitation. Thesemore » studies establish that the RRM domain of RBPMS forms a symmetrical dimer in the free state, with each monomer binding sequence-specifically to all three nucleotides of a CAC segment in the RNA bound state. Structure-guided mutations within the dimerization and RNA-binding interfaces of RBPMS RRM on RNA complex formation resulted in both disruption of dimerization and a decrease in RNA-binding affinity as observed by size exclusion chromatography and isothermal titration calorimetry. As anticipated from biochemical binding studies, over-expression of dimerization or RNA-binding mutants of Flag-HA-tagged RBPMS were no longer able to track with stress granules in HEK293 cells, thereby documenting the deleterious effects of such mutationsin vivo.« less

Astronaut Harrison Schmitt next to deployed U.S. flag on lunar surface

NASA Image and Video Library

1972-12-13

AS17-134-20384 (7-19 Dec. 1972) --- Scientist-astronaut Harrison H. Schmitt, lunar module pilot, is photographed next to the deployed United States flag during lunar surface extravehicular activity (EVA) at the Taurus-Littrow landing site. The highest part of the flag appears to point toward our planet Earth in the distant background. This picture was taken by astronaut Eugene A. Cernan, Apollo 17 commander. While astronauts Cernan and Schmitt descended in the Lunar Module (LM) to explore the moon, astronaut Ronald E. Evans, command module pilot, remained with the Command and Service Modules (CSM) in lunar orbit.

KSC-02pp1743

NASA Image and Video Library

2002-11-10

KENNEDY SPACE CENTER, FLA. -- Seminole Native American Veterans serve as color guard during a pre-launch Native American ceremony at the Rocket Garden in the KSC Visitor Complex. David Nunez, U.S. Navy, carries the State of Florida Flag; David Stephen Bowers, U.S. Army, carries the Flag of the United States of America; Charles Billie Hiers, U.S. Marine Corps., carries the Seminole Tribe of Florida Flag. The ceremony was part of several days' activities commemorating John B. Herrington as the first tribally enrolled Native American astronaut to fly on a Shuttle mission. Herrington is a Mission Specialist on STS-113.

An Evidence-Based Case Study of Unilateral Shin Splints: Do Red Flags Function in Paediatric Osteosarcoma?

PubMed

Rankin, Anne

2015-01-01

Physiotherapists use red flags to screen for serious pathology. Paediatric osteosarcoma is a rare disease, occurring predominantly in the area of the knee and shoulder, and it is not always included by physiotherapists on a differential diagnosis list. Traditional red flags do not always correspond to the initial signs and symptoms of osteosarcoma. Physiotherapists should routinely palpate along the length of the bone to detect a potential mass. The detection of a mass or symptoms that do not follow the expected course indicates the need for reassessment and possibly referral for further investigation.

KSC-07pd0767

NASA Image and Video Library

2007-04-02

KENNEDY SPACE CENTER, FLA. -- The NASA logo shines in the morning sun on the side of the Vehicle Assembly Building after completion of its repainting. The logo, which is known as the "meatball," measures 110 feet by 132 feet, or about 12,300 square feet. The American flag was also painted on the side of the VAB. The flag spans an area 209 feet by 110 feet, or about 23, 437 square feet. Each stripe is 9 feet wide and each star is 6 feet in diameter. The flag and logo were last painted in 1998, honoring NASA's 40th anniversary. Photo credit: NASA/Jim Grossmann

The Field-tested Learning Assessment Guide (FLAG): A Community Repository of Proven Alternative Assessment Instruments for STEM Education

NASA Astrophysics Data System (ADS)

Zeilik, M.; Garvin-Doxas, K.

2003-12-01

FLAG, the Field-tested Learning Assessment Guide (http://www.flaguide.org/) is a NSF funded website that offers broadly-applicable, self-contained modular classroom assessment techniques (CATs) and discipline-specific tools for STEM instructors creating new approaches to evaluate student learning, attitudes and performance. In particular, the FLAG contains proven techniques for alterative assessments---those needed for reformed, innovative STEM courses. Each tool has been developed, tested and refined in real classrooms at colleges and universities. The FLAG also contains an assessment primer, a section to help you select the most appropriate assessment technique(s) for your course goals, and other resources. In addition to references on instrument development and field-tested instruments on attitudes towards science, the FLAG also includes discipline-specific tools in Physics, Astronomy, Biology, and Mathematics. Building of the Geoscience collection is currently under way with the development of an instrument for detecting misconceptions of incoming freshmen on Space Science, which is being developed with the help of the Committee on Space Science and Astronomy of the American Association of Physics Teachers. Additional field-tested resources from the Geosciences are solicited from the community. Contributions should be sent to Michael Zeilik, zeilik@la.unm.edu. This work has been supported in part by NSF grant DUE 99-81155.

Ground squirrel tail-flag displays alter both predatory strike and ambush site selection behaviours of rattlesnakes

PubMed Central

Barbour, Matthew A.; Clark, Rulon W.

2012-01-01

Many species approach, inspect and signal towards their predators. These behaviours are often interpreted as predator-deterrent signals—honest signals that indicate to a predator that continued hunting is likely to be futile. However, many of these putative predator-deterrent signals are given when no predator is present, and it remains unclear if and why such signals deter predators. We examined the effects of one such signal, the tail-flag display of California ground squirrels, which is frequently given both during and outside direct encounters with northern Pacific rattlesnakes. We video-recorded and quantified the ambush foraging responses of rattlesnakes to tail-flagging displays from ground squirrels. We found that tail-flagging deterred snakes from striking squirrels, most likely by advertising squirrel vigilance (i.e. readiness to dodge a snake strike). We also found that tail-flagging by adult squirrels increased the likelihood that snakes would leave their ambush site, apparently by elevating the vigilance of nearby squirrels which reduces the profitability of the ambush site. Our results provide some of the first empirical evidence of the mechanisms by which a prey display, although frequently given in the absence of a predator, may still deter predators during encounters. PMID:22787023

OAS :: Member States

Science.gov Websites

General Assembly Governance H Human Development Human Rights I Indigenous Peoples Integral Development Scholarships School of Governance Science and Technology Social Development Summits of the Americas Sustainable (Commonwealth of) Dominica (Commonwealth of) Flag Dominican Republic Dominican Republic Flag Ecuador Ecuador

SERPent: Automated reduction and RFI-mitigation software for e-MERLIN

NASA Astrophysics Data System (ADS)

Peck, Luke W.; Fenech, Danielle M.

2013-08-01

The Scripted E-merlin Rfi-mitigation PipelinE for iNTerferometry (SERPent) is an automated reduction and RFI-mitigation procedure utilising the SumThreshold methodology (Offringa et al., 2010a), originally developed for the LOFAR pipeline. SERPent is written in the Parseltongue language enabling interaction with the Astronomical Image Processing Software (AIPS) program. Moreover, SERPent is a simple 'out of the box' Python script, which is easy to set up and is free of compilers. In addition to the flagging of RFI affected visibilities, the script also flags antenna zero-amplitude dropouts and Lovell telescope phase calibrator stationary scans inherent to the e-MERLIN system. Both the flagging and computational performances of SERPent are presented here, for e-MERLIN commissioning datasets for both L-band (1.3-1.8 GHz) and C-band (4-8 GHz) observations. RFI typically amounts to <20%-25% for the more problematic L-band observations and <5% for the generally RFI quieter C-band. The level of RFI detection and flagging is more accurate and delicate than visual manual flagging, with the output immediately ready for AIPS calibration. SERPent is fully parallelised and has been tested on a range of computing systems. The current flagging rate is at 110 GB day-1 on a 'high-end' computer (16 CPUs, 100 GB memory) which amounts to ˜6.9 GB CPU-1 day-1, with an expected increase in performance when e-MERLIN has completed its commissioning. The refining of automated reduction and calibration procedures is essential for the e-MERLIN legacy projects and future interferometers such as the SKA and the associated pathfinders (MeerKAT and ASKAP), where the vast data sizes (>TB) make traditional astronomer interactions unfeasible.

Official portrait of Astronaut Anna L. Fisher

NASA Technical Reports Server (NTRS)

1985-01-01

Official portrait of Astronaut Anna L. Fisher. Fisher is posing with her helmet on the table in front of her and the American flag appears over the opposite shoulder (34357); Posing with an empty table in front of her and the American flag behind her (34358).

14 CFR 121.535 - Responsibility for operational control: Flag operations.

Code of Federal Regulations, 2013 CFR

2013-01-01

... dispatcher are jointly responsible for the preflight planning, delay, and dispatch release of a flight in... AND OPERATIONS OPERATING REQUIREMENTS: DOMESTIC, FLAG, AND SUPPLEMENTAL OPERATIONS Flight Operations...— (1) Monitoring the progress of each flight; (2) Issuing necessary instructions and information for...

14 CFR 121.535 - Responsibility for operational control: Flag operations.

Code of Federal Regulations, 2011 CFR

2011-01-01

... dispatcher are jointly responsible for the preflight planning, delay, and dispatch release of a flight in... AND OPERATIONS OPERATING REQUIREMENTS: DOMESTIC, FLAG, AND SUPPLEMENTAL OPERATIONS Flight Operations...— (1) Monitoring the progress of each flight; (2) Issuing necessary instructions and information for...

14 CFR 121.535 - Responsibility for operational control: Flag operations.

Code of Federal Regulations, 2010 CFR

2010-01-01

... dispatcher are jointly responsible for the preflight planning, delay, and dispatch release of a flight in... AND OPERATIONS OPERATING REQUIREMENTS: DOMESTIC, FLAG, AND SUPPLEMENTAL OPERATIONS Flight Operations...— (1) Monitoring the progress of each flight; (2) Issuing necessary instructions and information for...

14 CFR 121.535 - Responsibility for operational control: Flag operations.

Code of Federal Regulations, 2012 CFR

2012-01-01

... dispatcher are jointly responsible for the preflight planning, delay, and dispatch release of a flight in... AND OPERATIONS OPERATING REQUIREMENTS: DOMESTIC, FLAG, AND SUPPLEMENTAL OPERATIONS Flight Operations...— (1) Monitoring the progress of each flight; (2) Issuing necessary instructions and information for...

14 CFR 121.535 - Responsibility for operational control: Flag operations.

Code of Federal Regulations, 2014 CFR

2014-01-01

... dispatcher are jointly responsible for the preflight planning, delay, and dispatch release of a flight in... AND OPERATIONS OPERATING REQUIREMENTS: DOMESTIC, FLAG, AND SUPPLEMENTAL OPERATIONS Flight Operations...— (1) Monitoring the progress of each flight; (2) Issuing necessary instructions and information for...

The Savannah River Site`s groundwater monitoring program. Third quarter 1990

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

Not Available

1991-05-06

The Environmental Protection Department/Environmental Monitoring Section (EPD/EMS) administers the Savannah River Site`s (SRS) Groundwater Monitoring Program. During third quarter 1990 (July through September) EPD/EMS conducted routine sampling of monitoring wells and drinking water locations. EPD/EMS established two sets of flagging criteria in 1986 to assist in the management of sample results. The flagging criteria do not define contamination levels; instead they aid personnel in sample scheduling, interpretation of data, and trend identification. The flagging criteria are based on detection limits, background levels in SRS groundwater, and drinking water standards. All analytical results from third quarter 1990 are listed in thismore » report, which is distributed to all site custodians. One or more analytes exceeded Flag 2 in 87 monitoring well series. Analytes exceeded Flat 2 for the first since 1984 in 14 monitoring well series. In addition to groundwater monitoring, EPD/EMS collected drinking water samples from SRS drinking water systems supplied by wells. The drinking water samples were analyzed for radioactive constituents.« less

The Savannah River Site's groundwater monitoring program

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

Not Available

1991-05-06

The Environmental Protection Department/Environmental Monitoring Section (EPD/EMS) administers the Savannah River Site's (SRS) Groundwater Monitoring Program. During third quarter 1990 (July through September) EPD/EMS conducted routine sampling of monitoring wells and drinking water locations. EPD/EMS established two sets of flagging criteria in 1986 to assist in the management of sample results. The flagging criteria do not define contamination levels; instead they aid personnel in sample scheduling, interpretation of data, and trend identification. The flagging criteria are based on detection limits, background levels in SRS groundwater, and drinking water standards. All analytical results from third quarter 1990 are listed in thismore » report, which is distributed to all site custodians. One or more analytes exceeded Flag 2 in 87 monitoring well series. Analytes exceeded Flat 2 for the first since 1984 in 14 monitoring well series. In addition to groundwater monitoring, EPD/EMS collected drinking water samples from SRS drinking water systems supplied by wells. The drinking water samples were analyzed for radioactive constituents.« less

Activities commemorating John B. Herrington as first Native American astronaut

NASA Technical Reports Server (NTRS)

2002-01-01

KENNEDY SPACE CENTER, FLA. -- Seminole Native American Veterans serve as color guard during a pre-launch Native American ceremony at the Rocket Garden in the KSC Visitor Complex. David Nunez, U.S. Navy, carries the State of Florida Flag; David Stephen Bowers, U.S. Army, carries the Flag of the United States of America; Charles Billie Hiers, U.S. Marine Corps., carries the Seminole Tribe of Florida Flag. The ceremony was part of several days' activities commemorating John B. Herrington as the first tribally enrolled Native American astronaut to fly on a Shuttle mission. Herrington is a Mission Specialist on STS-113.

An Evidence-Based Case Study of Unilateral Shin Splints: Do Red Flags Function in Paediatric Osteosarcoma?

PubMed Central

2015-01-01

ABSTRACT Physiotherapists use red flags to screen for serious pathology. Paediatric osteosarcoma is a rare disease, occurring predominantly in the area of the knee and shoulder, and it is not always included by physiotherapists on a differential diagnosis list. Traditional red flags do not always correspond to the initial signs and symptoms of osteosarcoma. Physiotherapists should routinely palpate along the length of the bone to detect a potential mass. The detection of a mass or symptoms that do not follow the expected course indicates the need for reassessment and possibly referral for further investigation. PMID:27504036

KSC-07pd0114

NASA Image and Video Library

2007-01-17

KENNEDY SPACE CENTER, FLA. -- On platforms suspended from the top of the 525-foot-high VAB, workers use rollers and brushes to repaint the U.S. flag on the southwest side of the Vehicle Assembly Building. The flag spans an area 209 feet by 110 feet, or about 23, 437 square feet. Each stripe is 9 feet wide and each star is 6 feet in diameter. The logo is also being painted. Known as the "meatball," the logo measures 110 feet by 132 feet, or about 12,300 square feet. The flag and logo were last painted in 1998, honoring NASA's 40th anniversary. Photo credit: NASA/George Shelton

KSC-07pd0115

NASA Image and Video Library

2007-01-17

KENNEDY SPACE CENTER, FLA. -- On platforms suspended from the top of the 525-foot-high VAB, workers use rollers and brushes to repaint the U.S. flag on the southwest side of the Vehicle Assembly Building. The flag spans an area 209 feet by 110 feet, or about 23, 437 square feet. Each stripe is 9 feet wide and each star is 6 feet in diameter. The logo is also being painted. Known as the "meatball," the logo measures 110 feet by 132 feet, or about 12,300 square feet. The flag and logo were last painted in 1998, honoring NASA's 40th anniversary. Photo credit: NASA/George Shelton

KSC-2011-1065

NASA Image and Video Library

2011-01-13

CAPE CANAVERAL, Fla. -- In accordance with a Presidential Proclamation, the American Flag in the Launch Complex 39 area of NASA's Kennedy Space Center in Florida flies at half-staff to honor U.S. Rep. Gabrielle Giffords and others who were wounded or killed during a shooting spree in Tucson, Arizona. This flag and all American Flags at U.S. military and government installations across the country and abroad will remain at half-staff until sunset on Jan. 14. Giffords is the wife of NASA astronaut Mark Kelly, who is scheduled to command space shuttle Endeavour's last mission, STS-134, to the International Space Station later this year. Photo credit: NASA/Frankie Martin

KSC-2011-1064

NASA Image and Video Library

2011-01-13

CAPE CANAVERAL, Fla. -- In accordance with a Presidential Proclamation, the American Flag in the Launch Complex 39 area of NASA's Kennedy Space Center in Florida flies at half-staff to honor U.S. Rep. Gabrielle Giffords and others who were wounded or killed during a shooting spree in Tucson, Arizona. This flag and all American Flags at U.S. military and government installations across the country and abroad will remain at half-staff until sunset on Jan. 14. Giffords is the wife of NASA astronaut Mark Kelly, who is scheduled to command space shuttle Endeavour's last mission, STS-134, to the International Space Station later this year. Photo credit: NASA/Frankie Martin

KSC-2011-1066

NASA Image and Video Library

2011-01-13

CAPE CANAVERAL, Fla. -- In accordance with a Presidential Proclamation, the American Flag in the Launch Complex 39 area of NASA's Kennedy Space Center in Florida flies at half-staff to honor U.S. Rep. Gabrielle Giffords and others who were wounded or killed during a shooting spree in Tucson, Arizona. This flag and all American Flags at U.S. military and government installations across the country and abroad will remain at half-staff until sunset on Jan. 14. Giffords is the wife of NASA astronaut Mark Kelly, who is scheduled to command space shuttle Endeavour's last mission, STS-134, to the International Space Station later this year. Photo credit: NASA/Frankie Martin

KSC-07pd0113

NASA Image and Video Library

2007-01-17

KENNEDY SPACE CENTER, FLA. -- On platforms suspended from the top of the 525-foot-high VAB, workers use rollers and brushes to repaint the U.S. flag on the southwest side of the Vehicle Assembly Building. The flag spans an area 209 feet by 110 feet, or about 23, 437 square feet. Each stripe is 9 feet wide and each star is 6 feet in diameter. The logo is also being painted. Known as the "meatball," the logo measures 110 feet by 132 feet, or about 12,300 square feet. The flag and logo were last painted in 1998, honoring NASA's 40th anniversary. Photo credit: NASA/George Shelton

KSC-07pd0039

NASA Image and Video Library

2007-01-09

KENNEDY SPACE CENTER, FLA. -- Painters' platforms are seen hanging on the side of Kennedy Space Center's 525-foot-high Vehicle Assembly Building to facilitate the repainting of the American flag and the NASA logo. Workers use rollers and brushes to do the painting. The flag and logo were last painted in 1998, honoring NASA's 40th anniversary. The flag spans an area 209 feet by 110 feet, or about 23,437 square feet. Each stripe is 9 feet wide and each star is 6 feet in diameter. The logo, also known as the "meatball," measures 110 feet by 132 feet, or about 12,300 square feet. Photo credit: NASA/Kim Shiflett

KSC-07pd0040

NASA Image and Video Library

2007-01-09

KENNEDY SPACE CENTER, FLA. -- Painters' platforms are seen hanging on the side of Kennedy Space Center's 525-foot-high Vehicle Assembly Building to facilitate the repainting of the American flag. The NASA logo is also being repainted. Workers use rollers and brushes to do the painting. The flag and logo were last painted in 1998, honoring NASA's 40th anniversary. The flag spans an area 209 feet by 110 feet, or about 23,437 square feet. Each stripe is 9 feet wide and each star is 6 feet in diameter. The logo, also known as the "meatball," measures 110 feet by 132 feet, or about 12,300 square feet. Photo credit: NASA/Kim Shiflett

KSC-07pd0116

NASA Image and Video Library

2007-01-17

KENNEDY SPACE CENTER, FLA. -- On platforms suspended from the top of the 525-foot-high VAB, workers use rollers and brushes to repaint the NASA logo on the southeast side of the Vehicle Assembly Building. Known as the "meatball," the logo measures 110 feet by 132 feet, or about 12,300 square feet. The U.S. flag is also being repainted. The flag spans an area 209 feet by 110 feet, or about 23, 437 square feet. Each stripe is 9 feet wide and each star is 6 feet in diameter. The flag and logo were last painted in 1998, honoring NASA's 40th anniversary. Photo credit: NASA/George Shelton

The nuclear form of glutathione peroxidase 4 is associated with sperm nuclear matrix and is required for proper paternal chromatin decondensation at fertilization.

PubMed

Puglisi, Rossella; Maccari, Irene; Pipolo, Simona; Conrad, Marcus; Mangia, Franco; Boitani, Carla

2012-04-01

The nuclear isoform of the selenoprotein Phospholipid Hydroperoxide Glutathione Peroxidase (nGPx4) is expressed in haploid male germ cells, contains several cysteines and is able to oxidize protein thiols, besides glutathione. In this study we have investigated the subnuclear localization of this isoform in isolated mouse male germ cells at different steps of maturation. Immunoblotting and confocal microscopy analyses of subnuclear fractions showed that nGPx4 is localized to the nuclear matrix together with well known markers of this subnuclear compartment like lamin B and topoisomerase IIβ at all stages of germ cell differentiation. The peculiar nGPx4 distribution was confirmed by both biochemical and morphological analyses of COS-1 cells overexpressing Flag-tagged nGPx4. To test the functional role of nGPx4 in the process of chromatin assembly, sperm isolated from the caput and the cauda epididymides of wild-type (WT) and genetically deficient in nGPx4 (nGPx4-KO) mice were analyzed in an in vitro chromatin decondensation assay. Results showed that sperm from nGPx4-KO mice were more prone to decondense than those from WT mice at all stages of epididymal maturation, providing conclusive evidence that nGPx4 is required for a correct sperm chromatin compaction. We next addressed the issue of whether the lack of nGPx4 impacts on early events occurring at fertilization. Indeed, in vitro fertilization experiments showed an acceleration of sperm chromatin dispersion in oocytes fertilized by nGpx4-KO sperm compared with control. Overall these data indicate that the absence of nGPx4 leads to sperm nuclear matrix/chromatin instability that may negatively affect the embryo development. Copyright © 2011 Wiley Periodicals, Inc.

High-level production and purification in a functional state of an extrasynaptic gamma-aminobutyric acid type A receptor containing α4β3δ subunits.

PubMed

Zhou, Xiaojuan; Desai, Rooma; Zhang, Yinghui; Stec, Wojciech J; Miller, Keith W; Jounaidi, Youssef

2018-01-01

The inhibitory γ-aminobutyric acid type A receptors are implicated in numerous physiological processes, including cognition and inhibition of neurotransmission, rendering them important molecular targets for many classes of drugs. Functionally, the entire GABAAR family of receptors can be subdivided into phasic, fast acting synaptic receptors, composed of α-, β- and γ-subunits, and tonic extrasynaptic receptors, many of which contain the δ-subunit in addition to α- and β-subunits. Whereas the subunit arrangement of the former group is agreed upon, that of the αβδ GABAARs remains unresolved by electrophysiological and pharmacological research. To resolve such issues will require biophysical techniques that demand quantities of receptor that have been previously unavailable. Therefore, we have engineered a stable cell line with tetracycline inducible expression of human α4-, β3- and N-terminally Flag-tagged δ-subunits. This cell line achieved a specific activity between 15 and 20 pmol [3H]muscimol sites/mg of membrane protein, making it possible to obtain 1 nmole of purified α4β3δ GABAAR from sixty 15-cm culture dishes. When induced, these cells exhibited agonist-induced currents with characteristics comparable to those previously reported for this receptor and a pharmacology that included strong modulation by etomidate and the δ-subunit-specific ligand, DS2. Immunoaffinity purification and reconstitution in CHAPS/asolectin micelles resulted in the retention of equilibrium allosteric interactions between the separate agonist, anesthetic and DS2 sites. Moreover, all three subunits retained glycosylation. The establishment of this well-characterized cell line will allow molecular level studies of tonic receptors to be undertaken.

What causes flag smut of wheat?

USDA-ARS?s Scientific Manuscript database

The causal agent of flag smut of wheat is currently subject to strict quarantine regulations in many countries and is believed to have a wide host range on wild and cultivated grasses. This fungus has been classified as both Urocystis agropyri and Urocystis tritici. Urocystis agropyri was first desc...

78 FR 63233 - National Offshore Safety Advisory Committee; Meeting

Federal Register 2010, 2011, 2012, 2013, 2014

2013-10-23

... Equipment in Hazardous Areas on Foreign Flag Mobile Offshore Drilling Units. (4) Safety Impact of Liftboat... Equipment in Hazardous Areas on Foreign Flag Mobile Offshore Drilling Units (MODUs); (d) Safety Impact of... DEPARTMENT OF HOMELAND SECURITY Coast Guard [Docket No. USCG-2013-0886] National Offshore Safety...

Fermilab Today

Science.gov Websites

Fermilab Today Tuesday, March 5, 2013 spacer Subscribe | Contact Us | Archive | Classifieds | Guidelines | Help Search GO spacer Calendar Have a safe day! Tuesday, March 5 3:30 p.m. DIRECTOR'S COFFEE Current Flag Status Flags at full staff Wilson Hall Cafe Tuesday, March 5 - Breakfast: All-American

China Report RED FLAG No 10, 16 MAY 1986

DTIC Science & Technology

1986-07-03

4U Create a New Situation in Building the Legal System for Science and Technology in Our Country (pp 26-29) (Wu Mingyu ) The New Technological...Beijing RED FLAG in Chinese No 10, 16 May 86 pp 26-29 [Article by Wu Mingyu [0702 2494 3842

The Ultimate Flag Games.

ERIC Educational Resources Information Center

Angel, Kenny; Sutton, Nancy

This paper describes six Ultimate Flag Games which offer a change from traditional games and sports that are usually geared toward athletically inclined students. These new games, aimed at middle school through college students, allow for success from the least-skilled through the most athletically talented students. Players are ability grouped…

75 FR 52722 - Taking and Importing of Marine Mammals

Federal Register 2010, 2011, 2012, 2013, 2014

2010-08-27

... International Dolphin Conservation Program (IDCP) by Spanish flag purse seine vessels or purse seine vessels operating under Spanish jurisdiction to be imported into the United States. The affirmative finding was... ETP by Spanish- flag purse seine vessels or purse seine vessels operating under Spanish jurisdiction...

33 CFR 334.850 - Lake Erie, west end, north of Erie Ordnance Depot, Lacarne, Ohio.

Code of Federal Regulations, 2010 CFR

2010-07-01

... firing into Area I, red flags will be flown from the safety tower at Camp Perry, and from flag poles in... fish will be discharged into the waters of the areas. (i) The regulations in this section shall be...

Race Discourse and the US Confederate Flag

ERIC Educational Resources Information Center

Holyfield, Lori; Moltz, Matthew Ryan; Bradley, Mindy S.

2009-01-01

Research reveals that racial hierarchies and "color-blind" racism is maintained through discourse. The current study utilizes exploratory data from focus groups in a predominantly white southern university in the United States to examine race talk, the Confederate Flag, and the construction of southern white identity. Drawing from…

Phloem Transport Of Arsenic Species From Flag Leaf To Grain During Grain Filling

EPA Science Inventory

Strategies to reduce arsenic (As) in rice grain, below concentrations that represent a serious human health concern, require that the mechanisms of As accumulation within grain be established. Therefore, retranslocation of As species from flag leaves into filling rice grain was ...

14 CFR 121.687 - Dispatch release: Flag and domestic operations.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 14 Aeronautics and Space 3 2014-01-01 2014-01-01 false Dispatch release: Flag and domestic operations. 121.687 Section 121.687 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF...) The dispatch release must contain, or have attached to it, weather reports, available weather...

14 CFR 121.687 - Dispatch release: Flag and domestic operations.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 14 Aeronautics and Space 3 2013-01-01 2013-01-01 false Dispatch release: Flag and domestic operations. 121.687 Section 121.687 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF...) The dispatch release must contain, or have attached to it, weather reports, available weather...

14 CFR 121.687 - Dispatch release: Flag and domestic operations.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 14 Aeronautics and Space 3 2012-01-01 2012-01-01 false Dispatch release: Flag and domestic operations. 121.687 Section 121.687 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF...) The dispatch release must contain, or have attached to it, weather reports, available weather...

14 CFR 121.687 - Dispatch release: Flag and domestic operations.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 14 Aeronautics and Space 3 2011-01-01 2011-01-01 false Dispatch release: Flag and domestic operations. 121.687 Section 121.687 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF...) The dispatch release must contain, or have attached to it, weather reports, available weather...

14 CFR 121.687 - Dispatch release: Flag and domestic operations.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 14 Aeronautics and Space 3 2010-01-01 2010-01-01 false Dispatch release: Flag and domestic operations. 121.687 Section 121.687 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF...) The dispatch release must contain, or have attached to it, weather reports, available weather...

46 CFR 154.24 - Foreign flag vessel: IMO Certificate.

Code of Federal Regulations, 2011 CFR

2011-10-01

....24 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CERTAIN BULK DANGEROUS CARGOES SAFETY STANDARDS FOR SELF-PROPELLED VESSELS CARRYING BULK LIQUEFIED GASES General § 154.24 Foreign flag... Construction and Equipment of Ships Carrying Liquefied Gases in Bulk, 1975” is usually sufficient evidence of...

46 CFR 154.24 - Foreign flag vessel: IMO Certificate.

Code of Federal Regulations, 2013 CFR

2013-10-01

....24 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CERTAIN BULK DANGEROUS CARGOES SAFETY STANDARDS FOR SELF-PROPELLED VESSELS CARRYING BULK LIQUEFIED GASES General § 154.24 Foreign flag... Construction and Equipment of Ships Carrying Liquefied Gases in Bulk, 1975” is usually sufficient evidence of...

46 CFR 154.24 - Foreign flag vessel: IMO Certificate.

Code of Federal Regulations, 2012 CFR

2012-10-01

....24 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CERTAIN BULK DANGEROUS CARGOES SAFETY STANDARDS FOR SELF-PROPELLED VESSELS CARRYING BULK LIQUEFIED GASES General § 154.24 Foreign flag... Construction and Equipment of Ships Carrying Liquefied Gases in Bulk, 1975” is usually sufficient evidence of...

46 CFR 154.24 - Foreign flag vessel: IMO Certificate.

Code of Federal Regulations, 2014 CFR

2014-10-01

....24 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CERTAIN BULK DANGEROUS CARGOES SAFETY STANDARDS FOR SELF-PROPELLED VESSELS CARRYING BULK LIQUEFIED GASES General § 154.24 Foreign flag... Construction and Equipment of Ships Carrying Liquefied Gases in Bulk, 1975” is usually sufficient evidence of...

46 CFR 154.24 - Foreign flag vessel: IMO Certificate.

Code of Federal Regulations, 2010 CFR

2010-10-01

....24 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CERTAIN BULK DANGEROUS CARGOES SAFETY STANDARDS FOR SELF-PROPELLED VESSELS CARRYING BULK LIQUEFIED GASES General § 154.24 Foreign flag... Construction and Equipment of Ships Carrying Liquefied Gases in Bulk, 1975” is usually sufficient evidence of...

Sunglint Detection for Unmanned and Automated Platforms

PubMed Central

Garaba, Shungudzemwoyo Pascal; Schulz, Jan; Wernand, Marcel Robert; Zielinski, Oliver

2012-01-01

We present an empirical quality control protocol for above-water radiometric sampling focussing on identifying sunglint situations. Using hyperspectral radiometers, measurements were taken on an automated and unmanned seaborne platform in northwest European shelf seas. In parallel, a camera system was used to capture sea surface and sky images of the investigated points. The quality control consists of meteorological flags, to mask dusk, dawn, precipitation and low light conditions, utilizing incoming solar irradiance (ES) spectra. Using 629 from a total of 3,121 spectral measurements that passed the test conditions of the meteorological flagging, a new sunglint flag was developed. To predict sunglint conspicuous in the simultaneously available sea surface images a sunglint image detection algorithm was developed and implemented. Applying this algorithm, two sets of data, one with (having too much or detectable white pixels or sunglint) and one without sunglint (having least visible/detectable white pixel or sunglint), were derived. To identify the most effective sunglint flagging criteria we evaluated the spectral characteristics of these two data sets using water leaving radiance (LW) and remote sensing reflectance (RRS). Spectral conditions satisfying ‘mean LW (700–950 nm) < 2 mW·m−2·nm−1·Sr−1’ or alternatively ‘minimum RRS (700–950 nm) < 0.010 Sr−1’, mask most measurements affected by sunglint, providing an efficient empirical flagging of sunglint in automated quality control.

Dynamic Black-Level Correction and Artifact Flagging in the Kepler Data Pipeline

NASA Technical Reports Server (NTRS)

Clarke, B. D.; Kolodziejczak, J. J.; Caldwell, D. A.

2013-01-01

Instrument-induced artifacts in the raw Kepler pixel data include time-varying crosstalk from the fine guidance sensor (FGS) clock signals, manifestations of drifting moiré pattern as locally correlated nonstationary noise and rolling bands in the images which find their way into the calibrated pixel time series and ultimately into the calibrated target flux time series. Using a combination of raw science pixel data, full frame images, reverse-clocked pixel data and ancillary temperature data the Keplerpipeline models and removes the FGS crosstalk artifacts by dynamically adjusting the black level correction. By examining the residuals to the model fits, the pipeline detects and flags spatial regions and time intervals of strong time-varying blacklevel (rolling bands ) on a per row per cadence basis. These flags are made available to downstream users of the data since the uncorrected rolling band artifacts could complicate processing or lead to misinterpretation of instrument behavior as stellar. This model fitting and artifact flagging is performed within the new stand-alone pipeline model called Dynablack. We discuss the implementation of Dynablack in the Kepler data pipeline and present results regarding the improvement in calibrated pixels and the expected improvement in cotrending performances as a result of including FGS corrections in the calibration. We also discuss the effectiveness of the rolling band flagging for downstream users and illustrate with some affected light curves.

Galectin-3 Negatively Regulates Hippocampus-Dependent Memory Formation through Inhibition of Integrin Signaling and Galectin-3 Phosphorylation

PubMed Central

Chen, Yan-Chu; Ma, Yun-Li; Lin, Cheng-Hsiung; Cheng, Sin-Jhong; Hsu, Wei-Lun; Lee, Eminy H.-Y.

2017-01-01

Galectin-3, a member of the galectin protein family, has been found to regulate cell proliferation, inhibit apoptosis and promote inflammatory responses. Galectin-3 is also expressed in the adult rat hippocampus, but its role in learning and memory function is not known. Here, we found that contextual fear-conditioning training, spatial training or injection of NMDA into the rat CA1 area each dramatically decreased the level of endogenous galectin-3 expression. Overexpression of galectin-3 impaired fear memory, whereas galectin-3 knockout (KO) enhanced fear retention, spatial memory and hippocampal long-term potentiation. Galectin-3 was further found to associate with integrin α3, an association that was decreased after fear-conditioning training. Transfection of the rat CA1 area with small interfering RNA against galectin-3 facilitated fear memory and increased phosphorylated focal adhesion kinase (FAK) levels, effects that were blocked by co-transfection of the FAK phosphorylation-defective mutant Flag-FAKY397F. Notably, levels of serine-phosphorylated galectin-3 were decreased by fear conditioning training. In addition, blockade of galectin-3 phosphorylation at Ser-6 facilitated fear memory, whereas constitutive activation of galectin-3 at Ser-6 impaired fear memory. Interestingly galectin-1 plays a role in fear-memory formation similar to that of galectin-3. Collectively, our data provide the first demonstration that galectin-3 is a novel negative regulator of memory formation that exerts its effects through both extracellular and intracellular mechanisms. PMID:28744198

An unusual case of seed dispersal in an invasive aquatic; yellow flag iris (Iris pseudacorus)

USDA-ARS?s Scientific Manuscript database

Understanding reproductive mode of invasive plants can help managers plan more efficacious control. Invasive aquatics typically reproduce primarily through vegetative means. Yellow flag iris is an invasive plant species often growing as an emergent aquatic. There have been contradictory reports of i...

78 FR 16475 - Procurement List; Proposed Additions and Deletion

Federal Register 2010, 2011, 2012, 2013, 2014

2013-03-15

... Activity: Military Resale-Defense Commissary Agency (DeCA), Fort Lee, VA Coverage: C-List for the requirements of military commissaries and exchanges as aggregated by the Defense Commissary Agency. NSN: 7510-01-389-2262--Self Stick Rectangular Flag, ``Sign Here'', 1.0'' X 1.75'', Yellow Flags NPA...

La Estrella Perdida (The Missing Star).

ERIC Educational Resources Information Center

Cohen, Allen Stephen

This story in Spanish and English is written for elementary school children in grades 1-5. The main character of the story is a star that is part of the Puerto Rican flag. The star leaves the flag and goes on a journey that is illustrated with pen-and-ink drawings. (Author/NCR)

Modifying Flag Football for Gender Equitable Engagement in Secondary Schools

ERIC Educational Resources Information Center

Kahan, David

2008-01-01

Flag or touch football is a popular activity unit in American secondary physical education curricula. However, unlike other sports its stigmatization as a masculine-typed activity and frequent inequitable distribution of game play opportunities at the skill positions (e.g., receiver, quarterback) results in the marginalization of female…

FMRI of visual working memory in high school football players.

PubMed

Shenk, Trey E; Robinson, Meghan E; Svaldi, Diana O; Abbas, Kausar; Breedlove, Katherine M; Leverenz, Larry J; Nauman, Eric A; Talavage, Thomas M

2015-01-01

Visual working memory deficits have been observed in at-risk athletes. This study uses a visual N-back working memory functional magnetic resonance imaging task to longitudinally assess asymptomatic football athletes for abnormal activity. Athletes were increasingly "flagged" as the season progressed. Flagging may provide early detection of injury.

10 CFR 1002.1 - Purpose.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 10 Energy 4 2010-01-01 2010-01-01 false Purpose. 1002.1 Section 1002.1 Energy DEPARTMENT OF ENERGY (GENERAL PROVISIONS) OFFICIAL SEAL AND DISTINGUISHING FLAG General § 1002.1 Purpose. The purpose of this part is to describe the official seal and distinguishing flag of the Department of Energy, and to...

14 CFR 1221.106 - Establishment of the NASA Flag.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 14 Aeronautics and Space 5 2013-01-01 2013-01-01 false Establishment of the NASA Flag. 1221.106 Section 1221.106 Aeronautics and Space NATIONAL AERONAUTICS AND SPACE ADMINISTRATION THE NASA SEAL AND OTHER DEVICES, AND THE CONGRESSIONAL SPACE MEDAL OF HONOR NASA Seal, NASA Insignia, NASA Logotype, NASA...

14 CFR 1221.106 - Establishment of the NASA Flag.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 14 Aeronautics and Space 5 2010-01-01 2010-01-01 false Establishment of the NASA Flag. 1221.106 Section 1221.106 Aeronautics and Space NATIONAL AERONAUTICS AND SPACE ADMINISTRATION THE NASA SEAL AND OTHER DEVICES, AND THE CONGRESSIONAL SPACE MEDAL OF HONOR NASA Seal, NASA Insignia, NASA Logotype, NASA...

14 CFR 1221.106 - Establishment of the NASA Flag.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 14 Aeronautics and Space 5 2012-01-01 2012-01-01 false Establishment of the NASA Flag. 1221.106 Section 1221.106 Aeronautics and Space NATIONAL AERONAUTICS AND SPACE ADMINISTRATION THE NASA SEAL AND OTHER DEVICES, AND THE CONGRESSIONAL SPACE MEDAL OF HONOR NASA Seal, NASA Insignia, NASA Logotype, NASA...

14 CFR 1221.106 - Establishment of the NASA Flag.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 14 Aeronautics and Space 5 2011-01-01 2010-01-01 true Establishment of the NASA Flag. 1221.106 Section 1221.106 Aeronautics and Space NATIONAL AERONAUTICS AND SPACE ADMINISTRATION THE NASA SEAL AND OTHER DEVICES, AND THE CONGRESSIONAL SPACE MEDAL OF HONOR NASA Seal, NASA Insignia, NASA Logotype, NASA...

14 CFR 121.467 - Flight attendant duty period limitations and rest requirements: Domestic, flag, and supplemental...

Code of Federal Regulations, 2014 CFR

2014-01-01

... 14 Aeronautics and Space 3 2014-01-01 2014-01-01 false Flight attendant duty period limitations and rest requirements: Domestic, flag, and supplemental operations. 121.467 Section 121.467 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION (CONTINUED) AIR CARRIERS...

14 CFR 121.467 - Flight attendant duty period limitations and rest requirements: Domestic, flag, and supplemental...

Code of Federal Regulations, 2011 CFR

2011-01-01

... 14 Aeronautics and Space 3 2011-01-01 2011-01-01 false Flight attendant duty period limitations and rest requirements: Domestic, flag, and supplemental operations. 121.467 Section 121.467 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION (CONTINUED) AIR CARRIERS...

14 CFR 121.467 - Flight attendant duty period limitations and rest requirements: Domestic, flag, and supplemental...

Code of Federal Regulations, 2012 CFR

2012-01-01

... 14 Aeronautics and Space 3 2012-01-01 2012-01-01 false Flight attendant duty period limitations and rest requirements: Domestic, flag, and supplemental operations. 121.467 Section 121.467 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION (CONTINUED) AIR CARRIERS...

14 CFR 121.467 - Flight attendant duty period limitations and rest requirements: Domestic, flag, and supplemental...

Code of Federal Regulations, 2013 CFR

2013-01-01

... 14 Aeronautics and Space 3 2013-01-01 2013-01-01 false Flight attendant duty period limitations and rest requirements: Domestic, flag, and supplemental operations. 121.467 Section 121.467 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION (CONTINUED) AIR CARRIERS...

76 FR 1665 - Stakeholder Meetings Regarding the U.S.-Flag Great Lakes Fleet Revitalization Study; Correction

Federal Register 2010, 2011, 2012, 2013, 2014

2011-01-11

... DEPARTMENT OF TRANSPORTATION Maritime Administration [Docket No. MARAD-2010-0111] Stakeholder Meetings Regarding the U.S.-Flag Great Lakes Fleet Revitalization Study; Correction AGENCY: Maritime Administration, Department of Transportation. ACTION: Correction Notice. SUMMARY: On December 29, 2010, at 75 FR...

76 FR 20080 - Inventory of U.S.-Flag Launch Barges

Federal Register 2010, 2011, 2012, 2013, 2014

2011-04-11

... DEPARTMENT OF TRANSPORTATION Maritime Administration [Docket Number MARAD 2011 0030] Inventory of...: Inventory of U.S.-Flag Launch Barges. SUMMARY: The Maritime Administration is updating its inventory of U.S..._program/Launch_Barge_Program.htm . DATES: Any comments on this inventory should be submitted in writing to...

77 FR 50086 - Taking and Importing of Marine Mammals

Federal Register 2010, 2011, 2012, 2013, 2014

2012-08-20

... International Dolphin Conservation Program (IDCP) by Spanish-flag purse seine vessels or purse seine vessels operating under Spanish jurisdiction to be imported into the United States. The affirmative finding was... derived from yellowfin tuna harvested in the ETP by Spanish-flag purse seine vessels or purse seine...

76 FR 52319 - Taking and Importing of Marine Mammals

Federal Register 2010, 2011, 2012, 2013, 2014

2011-08-22

... International Dolphin Conservation Program (IDCP) by Spanish-flag purse seine vessels or purse seine vessels operating under Spanish jurisdiction to be imported into the United States. The affirmative finding was... States of yellowfin tuna and products derived from yellowfin tuna harvested in the ETP by Spanish-flag...

50 CFR 32.59 - Rhode Island.

Code of Federal Regulations, 2013 CFR

2013-10-01

... to, the use of flagging, bright eyes, tacks, and paint), cutting, and/or removal of trees or.... We prohibit hunters field dressing deer within 100 feet (30 m) of a road or trail. 22. We prohibit... to, the use of flagging, bright eyes, tacks, and paint), cutting, and/or removal of trees or...

50 CFR 32.59 - Rhode Island.

Code of Federal Regulations, 2014 CFR

2014-10-01

... to, the use of flagging, bright eyes, tacks, and paint), cutting, and/or removal of trees or.... We prohibit hunters field dressing deer within 100 feet (30 m) of a road or trail. 22. We prohibit... to, the use of flagging, bright eyes, tacks, and paint), cutting, and/or removal of trees or...

50 CFR 32.59 - Rhode Island.

Code of Federal Regulations, 2012 CFR

2012-10-01

... to, the use of flagging, bright eyes, tacks, and paint), cutting, and/or removal of trees or.... We prohibit hunters field dressing deer within 100 feet (30 m) of a road or trail. 22. We prohibit... to, the use of flagging, bright eyes, tacks, and paint), cutting, and/or removal of trees or...

48 CFR 47.405 - Contract clause.

Code of Federal Regulations, 2010 CFR

2010-10-01

... TRANSPORTATION Air Transportation by U.S.-Flag Carriers 47.405 Contract clause. The contracting officer shall insert the clause at 52.247-63, “Preference for U.S.-Flag Air Carriers, in solicitations and contracts whenever it is possible that U.S. Government-financed international air transportation of personnel (and...

15 CFR 971.701 - Criteria for safety of life and property at sea.

Code of Federal Regulations, 2010 CFR

2010-01-01

... REGULATIONS OF THE ENVIRONMENTAL DATA SERVICE DEEP SEABED MINING REGULATIONS FOR COMMERCIAL RECOVERY PERMITS... by present laws and regulations. The primary inspection statutes pertaining to United States flag... Motor Vessels, and Freight Vessels). United States flag vessels will be required to meet all applicable...

48 CFR 47.501 - Definitions.

Code of Federal Regulations, 2011 CFR

2011-10-01

... of bulk liquid cargoes such as liquid petroleum products, vegetable oils, and molasses. U.S.-flag... cargoes of dry bulk items or, when carried in deep tanks, bulk liquids such as petroleum and vegetable oils. Foreign-flag vessel means any vessel of foreign registry including vessels owned by U.S. citizens...

48 CFR 47.501 - Definitions.

Code of Federal Regulations, 2010 CFR

2010-10-01

... of bulk liquid cargoes such as liquid petroleum products, vegetable oils, and molasses. U.S.-flag... cargoes of dry bulk items or, when carried in deep tanks, bulk liquids such as petroleum and vegetable oils. Foreign-flag vessel means any vessel of foreign registry including vessels owned by U.S. citizens...

The Impact of Flagging on the Admission Process.

ERIC Educational Resources Information Center

Cahalan-Laitusis, Cara; Mandinach, Ellen B.; Camara, Wayne J.

2003-01-01

Study explored issues surrounding flagging test scores taken under non-standard conditions and how the admission process could better serve students with disabilities. Respondents to survey felt current system was not adequately serving subgroups of students, believing some non-disabled students were manipulating the system to gain an advantage on…

75 FR 66125 - Federal Land Managers' Air Quality Related Values Work Group (FLAG)

Federal Register 2010, 2011, 2012, 2013, 2014

2010-10-27

... DEPARTMENT OF THE INTERIOR National Park Service Federal Land Managers' Air Quality Related Values... Public Comments document. The Federal Land Managers' Air Quality Related Values Work Group (FLAG) was... (the Agencies), to evaluate air pollution effects on their air quality related values (AQRVs); and (2...

46 CFR 154.15 - U.S. flag vessel: Endorsement application.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 46 Shipping 5 2014-10-01 2014-10-01 false U.S. flag vessel: Endorsement application. 154.15 Section 154.15 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CERTAIN BULK DANGEROUS CARGOES SAFETY STANDARDS FOR SELF-PROPELLED VESSELS CARRYING BULK LIQUEFIED GASES General § 154.15 U.S...

46 CFR 153.16 - Requirements for foreign flag vessel permits.

Code of Federal Regulations, 2012 CFR

2012-10-01

... 46 Shipping 5 2012-10-01 2012-10-01 false Requirements for foreign flag vessel permits. 153.16 Section 153.16 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CERTAIN BULK DANGEROUS CARGOES SHIPS CARRYING BULK LIQUID, LIQUEFIED GAS, OR COMPRESSED GAS HAZARDOUS MATERIALS General § 153.16...

46 CFR 154.15 - U.S. flag vessel: Endorsement application.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 46 Shipping 5 2010-10-01 2010-10-01 false U.S. flag vessel: Endorsement application. 154.15 Section 154.15 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CERTAIN BULK DANGEROUS CARGOES SAFETY STANDARDS FOR SELF-PROPELLED VESSELS CARRYING BULK LIQUEFIED GASES General § 154.15 U.S...

46 CFR 154.15 - U.S. flag vessel: Endorsement application.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 46 Shipping 5 2011-10-01 2011-10-01 false U.S. flag vessel: Endorsement application. 154.15 Section 154.15 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CERTAIN BULK DANGEROUS CARGOES SAFETY STANDARDS FOR SELF-PROPELLED VESSELS CARRYING BULK LIQUEFIED GASES General § 154.15 U.S...

46 CFR 153.16 - Requirements for foreign flag vessel permits.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 46 Shipping 5 2011-10-01 2011-10-01 false Requirements for foreign flag vessel permits. 153.16 Section 153.16 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CERTAIN BULK DANGEROUS CARGOES SHIPS CARRYING BULK LIQUID, LIQUEFIED GAS, OR COMPRESSED GAS HAZARDOUS MATERIALS General § 153.16...

46 CFR 154.15 - U.S. flag vessel: Endorsement application.

Code of Federal Regulations, 2012 CFR

2012-10-01

... 46 Shipping 5 2012-10-01 2012-10-01 false U.S. flag vessel: Endorsement application. 154.15 Section 154.15 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CERTAIN BULK DANGEROUS CARGOES SAFETY STANDARDS FOR SELF-PROPELLED VESSELS CARRYING BULK LIQUEFIED GASES General § 154.15 U.S...

46 CFR 154.15 - U.S. flag vessel: Endorsement application.

Code of Federal Regulations, 2013 CFR

2013-10-01

... 46 Shipping 5 2013-10-01 2013-10-01 false U.S. flag vessel: Endorsement application. 154.15 Section 154.15 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CERTAIN BULK DANGEROUS CARGOES SAFETY STANDARDS FOR SELF-PROPELLED VESSELS CARRYING BULK LIQUEFIED GASES General § 154.15 U.S...

46 CFR 153.16 - Requirements for foreign flag vessel permits.

Code of Federal Regulations, 2013 CFR

2013-10-01

... 46 Shipping 5 2013-10-01 2013-10-01 false Requirements for foreign flag vessel permits. 153.16 Section 153.16 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CERTAIN BULK DANGEROUS CARGOES SHIPS CARRYING BULK LIQUID, LIQUEFIED GAS, OR COMPRESSED GAS HAZARDOUS MATERIALS General § 153.16...

46 CFR 153.16 - Requirements for foreign flag vessel permits.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 46 Shipping 5 2010-10-01 2010-10-01 false Requirements for foreign flag vessel permits. 153.16 Section 153.16 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CERTAIN BULK DANGEROUS CARGOES SHIPS CARRYING BULK LIQUID, LIQUEFIED GAS, OR COMPRESSED GAS HAZARDOUS MATERIALS General § 153.16...

46 CFR 153.16 - Requirements for foreign flag vessel permits.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 46 Shipping 5 2014-10-01 2014-10-01 false Requirements for foreign flag vessel permits. 153.16 Section 153.16 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CERTAIN BULK DANGEROUS CARGOES SHIPS CARRYING BULK LIQUID, LIQUEFIED GAS, OR COMPRESSED GAS HAZARDOUS MATERIALS General § 153.16...

Cardiac-specific overexpression of sarcolipin inhibits sarco(endo)plasmic reticulum Ca2+ ATPase (SERCA2a) activity and impairs cardiac function in mice

PubMed Central

Asahi, Michio; Otsu, Kinya; Nakayama, Hiroyuki; Hikoso, Shungo; Takeda, Toshihiro; Gramolini, Anthony O.; Trivieri, Maria G.; Oudit, Gavin Y.; Morita, Takashi; Kusakari, Yoichiro; Hirano, Shuta; Hongo, Kenichi; Hirotani, Shinichi; Yamaguchi, Osamu; Peterson, Alan; Backx, Peter H.; Kurihara, Satoshi; Hori, Masatsugu; MacLennan, David H.

2004-01-01

Sarcolipin (SLN) inhibits the cardiac sarco(endo)plasmic reticulum Ca2+ ATPase (SERCA2a) by direct binding and is superinhibitory if it binds through phospholamban (PLN). To determine whether overexpression of SLN in the heart might impair cardiac function, transgenic (TG) mice were generated with cardiac-specific overexpression of NF-SLN (SLN tagged at its N terminus with the FLAG epitope). The level of NF-SLN expression (the NF-SLN/PLN expression ratio) was equivalent to that which induces profound superinhibition when coexpressed with PLN and SERCA2a in HEK-293 cells. In TG hearts, the apparent affinity of SERCA2a for Ca2+ was decreased compared with non-TG littermate control hearts. Invasive hemodynamic and echocardiographic analyses revealed impaired cardiac contractility and ventricular hypertrophy in TG mice. Basal PLN phosphorylation was reduced. In isolated papillary muscle subjected to isometric tension, peak amplitudes of Ca2+ transients and peak tensions were reduced, whereas decay times of Ca2+ transients and relaxation times of tension were increased in TG mice. Isoproterenol largely restored contractility in papillary muscle and stimulated PLN phosphorylation to wild-type levels in intact hearts. No compensatory changes in expression of SERCA2a, PLN, ryanodine receptor, and calsequestrin were observed in TG hearts. Coimmunoprecipitation indicated that overexpressed NF-SLN was bound to both SERCA2a and PLN, forming a ternary complex. These data suggest that NF-SLN overexpression inhibits SERCA2a through stabilization of SERCA2a–PLN interaction in the absence of PLN phosphorylation and through the inhibition of PLN phosphorylation. Inhibition of SERCA2a impairs contractility and calcium cycling, but responsiveness to β-adrenergic agonists may prevent progression to heart failure. PMID:15201433

The Compleat TRAIDsman.

DTIC Science & Technology

1969-09-01

uses a COMMON block of 663 cells . PLOTT works from a table of data, and assigns to each trace (up to 6 traces) a preset character. Once yoor data is...ALFA 000777 1 ~L P M A LP A U VU I 00) 󈧑 EL4 ’ I’LAC 4-43 FLIER slU.-r t;IINk fLIE" (5,LP.KONUA IlEI 335 50C k k LJ~C I~ N’.12 5 ExTEN, REVISION of...FLAG, PLACLD IN CumM"O. 753 C N3.0,0 AS RUN OAIE (IF BLANK* TAKEN FROM SYSTEM C DATE CELL ) ?SS C e1-63 13 EXECUTION TIME LIMIT IN MINUTES (IF ?b6 C

Patterns of cytosine methylation in an elite rice hybrid and its parental lines, detected by a methylation-sensitive amplification polymorphism technique.

PubMed

Xiong, L Z; Xu, C G; Saghai Maroof, M A; Zhang, Q

1999-04-01

DNA methylation is known to play an important role in the regulation of gene expression in eukaryotes. In this study, we assessed the extent and pattern of cytosine methylation in the rice genome, using the technique of methylation-sensitive amplified polymorphism (MSAP), which is a modification of the amplified fragment length polymorphism (AFLP) method that makes use of the differential sensitivity of a pair of isoschizomers to cytosine methylation. The tissues assayed included seedlings and flag leaves of an elite rice hybrid, Shanyou 63, and the parental lines Zhenshan 97 and Minghui 63. In all, 1076 fragments, each representing a recognition site cleaved by either or both of the isoschizomers, were amplified using 16 pairs of selective primers. A total of 195 sites were found to be methylated at cytosines in one or both parents, and the two parents showed approximately the same overall degree of methylation (16.3%), as revealed by the incidence of differential digestion by the isoschizomers. Four classes of patterns were identified in a comparative assay of cytosine methylation in the parents and hybrid; increased methylation was detected in the hybrid compared to the parents at some of the recognition sites, while decreased methylation in the hybrid was detected at other sites. A small proportion of the sites was found to be differentially methylated in seedlings and flag leaves; DNA from young seedlings was methylated to a greater extent than that from flag leaves. Almost all of the methylation patterns detected by MSAP could be confirmed by Southern analysis using the isolated amplified fragments as probes. The results clearly demonstrate that the MSAP technique is highly efficient for large-scale detection of cytosine methylation in the rice genome. We believe that the technique can be adapted for use in other plant species.

CNNs flag recognition preprocessing scheme based on gray scale stretching and local binary pattern

NASA Astrophysics Data System (ADS)

Gong, Qian; Qu, Zhiyi; Hao, Kun

2017-07-01

Flag is a rather special recognition target in image recognition because of its non-rigid features with the location, scale and rotation characteristics. The location change can be handled well by the depth learning algorithm Convolutional Neural Networks (CNNs), but the scale and rotation changes are quite a challenge for CNNs. Since it has good rotation and gray scale invariance, the local binary pattern (LBP) is combined with grayscale stretching and CNNs to make LBP and grayscale stretching as CNNs pretreatment, which can not only significantly improve the efficiency of flag recognition, but can also evaluate the recognition effect through ROC, accuracy, MSE and quality factor.

KSC-07pd0014

NASA Image and Video Library

2007-01-04

KENNEDY SPACE CENTER, FLA. -- Elevated platforms are seen hanging on the side of Kennedy Space Center's Vehicle Assembly Building to facilitate the repainting of the American flag and NASA logo. Workers, suspended on the platforms from the top of the 525-foot-high VAB, use rollers and brushes to do the painting. The flag and logo were last painted in 1998, honoring NASA's 40th anniversary. The flag spans an area 209 feet by 110 feet, or about 23, 437 square feet. Each stripe is 9 feet wide and each star is 6 feet in diameter. The logo, also known as the "meatball," measures 110 feet by 132 feet, or about 12,300 square feet. Photo credit: NASA/George Shelton

KSC-07pd0037

NASA Image and Video Library

2007-01-09

KENNEDY SPACE CENTER, FLA. -- Painters' scaffolding is seen hanging on the side of Kennedy Space Center's Vehicle Assembly Building to facilitate the repainting of the American flag. The NASA logo is also being repainted. Workers, suspended on platforms from the top of the 525-foot-high VAB, use rollers and brushes to do the painting. The flag and logo were last painted in 1998, honoring NASA's 40th anniversary. The flag spans an area 209 feet by 110 feet, or about 23,437 square feet. Each stripe is 9 feet wide and each star is 6 feet in diameter. The logo, also known as the "meatball," measures 110 feet by 132 feet, or about 12,300 square feet. Photo credit: NASA/Kim Shiflett

KSC-07pd0038

NASA Image and Video Library

2007-01-09

KENNEDY SPACE CENTER, FLA. -- Painters' scaffolding is seen hanging on the side of Kennedy Space Center's Vehicle Assembly Building to facilitate the repainting of the American flag and the NASA logo. Workers, suspended on platforms from the top of the 525-foot-high VAB, use rollers and brushes to do the painting. The flag and logo were last painted in 1998, honoring NASA's 40th anniversary. The flag spans an area 209 feet by 110 feet, or about 23,437 square feet. Each stripe is 9 feet wide and each star is 6 feet in diameter. The logo, also known as the "meatball," measures 110 feet by 132 feet, or about 12,300 square feet. Photo credit: NASA/Kim Shiflett

KSC-07pd0036

NASA Image and Video Library

2007-01-09

KENNEDY SPACE CENTER, FLA. -- Painters' scaffolding is seen hanging on the side of Kennedy Space Center's Vehicle Assembly Building to facilitate the repainting of the NASA logo. The American flag is also being repainted. Workers, suspended on platforms from the top of the 525-foot-high VAB, use rollers and brushes to do the painting. The flag and logo were last painted in 1998, honoring NASA's 40th anniversary. The flag spans an area 209 feet by 110 feet, or about 23,437 square feet. Each stripe is 9 feet wide and each star is 6 feet in diameter. The logo, also known as the "meatball," measures 110 feet by 132 feet, or about 12,300 square feet. Photo credit: NASA/Kim Shiflett

KSC-07pd0015

NASA Image and Video Library

2007-01-04

KENNEDY SPACE CENTER, FLA. -- Elevated platforms are seen hanging on the side of Kennedy Space Center's Vehicle Assembly Building to facilitate the repainting of the American flag and NASA logo. Workers, suspended on the platforms from the top of the 525-foot-high VAB, use rollers and brushes to do the painting. The flag and logo were last painted in 1998, honoring NASA's 40th anniversary. The flag spans an area 209 feet by 110 feet, or about 23, 437 square feet. Each stripe is 9 feet wide and each star is 6 feet in diameter. The logo, also known as the "meatball," measures 110 feet by 132 feet, or about 12,300 square feet. Photo credit: NASA/George Shelton

Workers painting the Flag and Meatball on the VAB

NASA Image and Video Library

2007-01-03

Elevated platforms are seen hanging on the side of Kennedy Space Center's Vehicle Assembly Building in a view looking across from the turn basin. To the right is the large external tank barge. Workers, suspended on the platforms from the top of the 525-foot-high VAB, use rollers and brushes to do the painting. The flag and logo were last painted in 1998, honoring NASA's 40th anniversary. The flag spans an area 209 feet by 110 feet, or about 23, 437 square feet. Each stripe is 9 feet wide and each star is 6 feet in diameter. The logo, also known as the "meatball," measures 110 feet by 132 feet, or about 12,300 square feet.

A general higher-order remap algorithm for ALE calculations

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

Chiravalle, Vincent P

2011-01-05

A numerical technique for solving the equations of fluid dynamics with arbitrary mesh motion is presented. The three phases of the Arbitrary Lagrangian Eulerian (ALE) methodology are outlined: the Lagrangian phase, grid relaxation phase and remap phase. The Lagrangian phase follows a well known approach from the HEMP code; in addition the strain rate andflow divergence are calculated in a consistent manner according to Margolin. A donor cell method from the SALE code forms the basis of the remap step, but unlike SALE a higher order correction based on monotone gradients is also added to the remap. Four test problemsmore » were explored to evaluate the fidelity of these numerical techniques, as implemented in a simple test code, written in the C programming language, called Cercion. Novel cell-centered data structures are used in Cercion to reduce the complexity of the programming and maximize the efficiency of memory usage. The locations of the shock and contact discontinuity in the Riemann shock tube problem are well captured. Cercion demonstrates a high degree of symmetry when calculating the Sedov blast wave solution, with a peak density at the shock front that is similar to the value determined by the RAGE code. For a flyer plate test problem both Cercion and FLAG give virtually the same velocity temporal profile at the target-vacuum interface. When calculating a cylindrical implosion of a steel shell, Cercion and FLAG agree well and the Cercion results are insensitive to the use of ALE.« less

Soccer; Speedball; Flag Football, June 1976--June 1978. NAGWS Guide.

ERIC Educational Resources Information Center

Messing, Anne, Ed.; And Others

This guide for soccer, speedball, and flag football is one in a series of guides for 22 sports published by the National Association for Girls and Women in Sport (NAGWS). Guides contain information on NAGWS-approved playing rules, officials' ratings, articles on teaching, coaching and organization, rules governing national championships,…

Defense.gov Special Report: Travels with Carter - May 2013

Science.gov Websites

Germany Germany Flag of Djibouti Djibouti Flag of Afghanistan Afghanistan Top Stories Carter Wraps Up , Civilians Breedlove Takes Charge at European Command Carter Arrives in Germany for Eucom Change of Command Meetings Carter Attends Eucom Change of Command Ceremony in Stuttgart, Germany Travel Locations Travel

No Experience Necessary: A Guide to Employment for the Female Liberal Arts Graduate.

ERIC Educational Resources Information Center

Friedman, Sande; Schwartz, Lois C.

A guide for beginning employment for female liberal arts graduates (referred to as FLAGs), this book describes 14 fields found to be presently most accessible to FLAGs without advanced degrees: advertising, the art world, banking and finance, book publishing, computer programing, fashion (and beauty), films, Government, magazine and newspaper…

Tips to Increase Girls' Participation in Flag Football Units

ERIC Educational Resources Information Center

Hannon, James C.; Ratliffe, Thomas

2006-01-01

Despite the apparent popularity of flag football as an activity in physical education class and football as an after-school offering for girls, studies related to gender stereotyping of sports have found overwhelming evidence indicating that football is perceived as a masculine activity among males and females in primary school, secondary school,…

Chemical Sciences and Engineering - US China Electric Vehicle and Battery

Science.gov Websites

Technology Workshop Argonne National Laboratory Chemical Sciences & Engineering DOE Logo Photo Gallery Hotels Maps Bus Schedule Contact Us TCS Building and Conference Center, Argonne National Lab TCS Building and Conference Center United States Flag China flag 2011 U.S.-China Electric Vehicle

22 CFR 228.22 - Air transportation.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 22 Foreign Relations 1 2011-04-01 2011-04-01 false Air transportation. 228.22 Section 228.22... for USAID Financing § 228.22 Air transportation. (a) The eligibility of air transportation is determined by the flag registry of the aircraft. The term “U.S. flag air carrier” means one of a class of air...

14 CFR 1221.113 - Use of the NASA Flags.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 14 Aeronautics and Space 5 2010-01-01 2010-01-01 false Use of the NASA Flags. 1221.113 Section 1221.113 Aeronautics and Space NATIONAL AERONAUTICS AND SPACE ADMINISTRATION THE NASA SEAL AND OTHER DEVICES, AND THE CONGRESSIONAL SPACE MEDAL OF HONOR NASA Seal, NASA Insignia, NASA Logotype, NASA Program...

14 CFR 1221.113 - Use of the NASA Flags.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 14 Aeronautics and Space 5 2011-01-01 2010-01-01 true Use of the NASA Flags. 1221.113 Section 1221.113 Aeronautics and Space NATIONAL AERONAUTICS AND SPACE ADMINISTRATION THE NASA SEAL AND OTHER DEVICES, AND THE CONGRESSIONAL SPACE MEDAL OF HONOR NASA Seal, NASA Insignia, NASA Logotype, NASA Program...

14 CFR 1221.113 - Use of the NASA Flags.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 14 Aeronautics and Space 5 2013-01-01 2013-01-01 false Use of the NASA Flags. 1221.113 Section 1221.113 Aeronautics and Space NATIONAL AERONAUTICS AND SPACE ADMINISTRATION THE NASA SEAL AND OTHER DEVICES, AND THE CONGRESSIONAL SPACE MEDAL OF HONOR NASA Seal, NASA Insignia, NASA Logotype, NASA Program...

14 CFR 1221.113 - Use of the NASA Flags.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 14 Aeronautics and Space 5 2012-01-01 2012-01-01 false Use of the NASA Flags. 1221.113 Section 1221.113 Aeronautics and Space NATIONAL AERONAUTICS AND SPACE ADMINISTRATION THE NASA SEAL AND OTHER DEVICES, AND THE CONGRESSIONAL SPACE MEDAL OF HONOR NASA Seal, NASA Insignia, NASA Logotype, NASA Program...

Defense.gov Special Report: Travels With Hagel

Science.gov Websites

. Flag of Mexico. Flag of Guatemala. April 2014 Defense Secretary Chuck Hagel began a three-day trip to troops who conduct military-to-military engagements in the region. He then visited Mexico City to attend the second U.S.-Canada-Mexico North American Defense Ministerial conference before heading to

49 CFR 214.521 - Flagging equipment for on-track roadway maintenance machines and hi-rail vehicles.

Code of Federal Regulations, 2010 CFR

2010-10-01

... maintenance machines and hi-rail vehicles. 214.521 Section 214.521 Transportation Other Regulations Relating... WORKPLACE SAFETY On-Track Roadway Maintenance Machines and Hi-Rail Vehicles § 214.521 Flagging equipment for on-track roadway maintenance machines and hi-rail vehicles. Each on-track roadway maintenance machine...

48 CFR 47.403-2 - Air transport agreements between the United States and foreign governments.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 48 Federal Acquisition Regulations System 1 2011-10-01 2011-10-01 false Air transport agreements....-Flag Carriers 47.403-2 Air transport agreements between the United States and foreign governments... attend, the use of a foreign-flag air carrier that provides transportation under an air transport...

14 CFR 121.551 - Restriction or suspension of operation: Domestic and flag operations.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 14 Aeronautics and Space 3 2010-01-01 2010-01-01 false Restriction or suspension of operation: Domestic and flag operations. 121.551 Section 121.551 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION... conditions, that are a hazard to safe operations, it shall restrict or suspend operations until those...

14 CFR 121.551 - Restriction or suspension of operation: Domestic and flag operations.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 14 Aeronautics and Space 3 2011-01-01 2011-01-01 false Restriction or suspension of operation: Domestic and flag operations. 121.551 Section 121.551 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION... conditions, that are a hazard to safe operations, it shall restrict or suspend operations until those...

38 CFR 1.10 - Eligibility for and disposition of the United States flag for burial purposes.

Code of Federal Regulations, 2010 CFR

2010-07-01

... separation from such service under conditions other than dishonorable, on or after April 25, 1951. (Authority... issued a flag on application signed by guardian). (iii) Parents, including adoptive, stepparents, and foster parents. (iv) Brothers or sisters, including brothers or sisters of the halfblood. (v) Uncles or...

50 CFR 32.49 - New Jersey.

Code of Federal Regulations, 2011 CFR

2011-10-01

... marking (this includes but is not limited to, the use of flagging, bright eyes, tacks, and paint), cutting... (this includes but is not limited to, the use of flagging, bright eyes, tacks, and paint), cutting, and..., Lily Lake, and the posted fishing areas along the south side of Parkertown Dock Road, North side of...

46 CFR 280.2 - Definitions.

Code of Federal Regulations, 2011 CFR

2011-10-01

.... For purposes of this part only: (a) Commercial cargo means cargo other than military cargo and civilian preference cargo. (b) Military cargo means that cargo required to be carried on a U.S.-flag vessel... by law to be carried on a U.S.-flag vessel, including, but not limited to, cargo required to be...

14 CFR 121.615 - Dispatch or flight release over water: Flag and supplemental operations.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 14 Aeronautics and Space 3 2011-01-01 2011-01-01 false Dispatch or flight release over water: Flag and supplemental operations. 121.615 Section 121.615 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION (CONTINUED) AIR CARRIERS AND OPERATORS FOR COMPENSATION OR HIRE: CERTIFICATION AND OPERATIONS OPERATING...

75 FR 51099 - Final Supplementary Rules for Public Land in Oregon and Washington

Federal Register 2010, 2011, 2012, 2013, 2014

2010-08-18

... must not operate a motorcycle or ATV/OHV without a safety flag on Peterson Road, Juniper Road, Smith Canyon Road, and/or Wilderness Road. Safety flags are not required for street-legal, four- wheeled... the public's health and safety. They provide needed guidance in the areas of special forest products...

12 CFR Appendix J to Part 571 - Interagency Guidelines on Identity Theft Detection, Prevention, and Mitigation

Code of Federal Regulations, 2013 CFR

2013-01-01

.... III. Detecting Red Flags The Program's policies and procedures should address the detection of Red... detect relevant Red Flags that may arise in the performance of the service provider's activities, and... or creditor detects a fraud or active duty alert; (c) Implementing any requirements for furnishers of...

12 CFR Appendix J to Part 171 - Interagency Guidelines on Identity Theft Detection, Prevention, and Mitigation

Code of Federal Regulations, 2014 CFR

2014-01-01

.... III. Detecting Red Flags The Program's policies and procedures should address the detection of Red... detect relevant Red Flags that may arise in the performance of the service provider's activities, and... or creditor detects a fraud or active duty alert; (c) Implementing any requirements for furnishers of...

12 CFR Appendix J to Part 171 - Interagency Guidelines on Identity Theft Detection, Prevention, and Mitigation

Code of Federal Regulations, 2012 CFR

2012-01-01

.... III. Detecting Red Flags The Program's policies and procedures should address the detection of Red... detect relevant Red Flags that may arise in the performance of the service provider's activities, and... or creditor detects a fraud or active duty alert; (c) Implementing any requirements for furnishers of...

12 CFR Appendix J to Part 571 - Interagency Guidelines on Identity Theft Detection, Prevention, and Mitigation

Code of Federal Regulations, 2012 CFR

2012-01-01

.... III. Detecting Red Flags The Program's policies and procedures should address the detection of Red... detect relevant Red Flags that may arise in the performance of the service provider's activities, and... or creditor detects a fraud or active duty alert; (c) Implementing any requirements for furnishers of...

12 CFR Appendix J to Part 571 - Interagency Guidelines on Identity Theft Detection, Prevention, and Mitigation

Code of Federal Regulations, 2014 CFR

2014-01-01

.... III. Detecting Red Flags The Program's policies and procedures should address the detection of Red... detect relevant Red Flags that may arise in the performance of the service provider's activities, and... or creditor detects a fraud or active duty alert; (c) Implementing any requirements for furnishers of...

12 CFR Appendix J to Part 571 - Interagency Guidelines on Identity Theft Detection, Prevention, and Mitigation

Code of Federal Regulations, 2011 CFR

2011-01-01

.... III. Detecting Red Flags The Program's policies and procedures should address the detection of Red... detect relevant Red Flags that may arise in the performance of the service provider's activities, and... or creditor detects a fraud or active duty alert; (c) Implementing any requirements for furnishers of...

12 CFR Appendix J to Part 171 - Interagency Guidelines on Identity Theft Detection, Prevention, and Mitigation

Code of Federal Regulations, 2013 CFR

2013-01-01

.... III. Detecting Red Flags The Program's policies and procedures should address the detection of Red... detect relevant Red Flags that may arise in the performance of the service provider's activities, and... or creditor detects a fraud or active duty alert; (c) Implementing any requirements for furnishers of...

49 CFR 214.521 - Flagging equipment for on-track roadway maintenance machines and hi-rail vehicles.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 49 Transportation 4 2011-10-01 2011-10-01 false Flagging equipment for on-track roadway maintenance machines and hi-rail vehicles. 214.521 Section 214.521 Transportation Other Regulations Relating to Transportation (Continued) FEDERAL RAILROAD ADMINISTRATION, DEPARTMENT OF TRANSPORTATION RAILROAD WORKPLACE SAFETY On-Track Roadway Maintenance...

49 CFR 214.521 - Flagging equipment for on-track roadway maintenance machines and hi-rail vehicles.

Code of Federal Regulations, 2012 CFR

2012-10-01

... 49 Transportation 4 2012-10-01 2012-10-01 false Flagging equipment for on-track roadway maintenance machines and hi-rail vehicles. 214.521 Section 214.521 Transportation Other Regulations Relating to Transportation (Continued) FEDERAL RAILROAD ADMINISTRATION, DEPARTMENT OF TRANSPORTATION RAILROAD WORKPLACE SAFETY On-Track Roadway Maintenance...

15 CFR 970.801 - Criteria for safety of life and property at sea.

Code of Federal Regulations, 2010 CFR

2010-01-01

... REGULATIONS OF THE ENVIRONMENTAL DATA SERVICE DEEP SEABED MINING REGULATIONS FOR EXPLORATION LICENSES Safety... inspection statutes pertaining to United States flag vessels are: 46 U.S.C. 86 (Loadlines); 46 U.S.C. 395... hire). All United States flag vessels will be required to meet existing regulatory requirements...

46 CFR 382.2 - Data submission.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 46 Shipping 8 2010-10-01 2010-10-01 false Data submission. 382.2 Section 382.2 Shipping MARITIME... CARRIAGE OF BULK AND PACKAGED PREFERENCE CARGOES ON U.S.-FLAG COMMERCIAL VESSELS § 382.2 Data submission... shall be performed on the basis of cost data provided by the U.S.-flag vessel operator, as specified...

46 CFR 382.2 - Data submission.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 46 Shipping 8 2011-10-01 2011-10-01 false Data submission. 382.2 Section 382.2 Shipping MARITIME... CARRIAGE OF BULK AND PACKAGED PREFERENCE CARGOES ON U.S.-FLAG COMMERCIAL VESSELS § 382.2 Data submission... shall be performed on the basis of cost data provided by the U.S.-flag vessel operator, as specified...

46 CFR 154.22 - Foreign flag vessel: Certificate of Compliance endorsement application.

Code of Federal Regulations, 2012 CFR

2012-10-01

... (CONTINUED) CERTAIN BULK DANGEROUS CARGOES SAFETY STANDARDS FOR SELF-PROPELLED VESSELS CARRYING BULK LIQUEFIED GASES General § 154.22 Foreign flag vessel: Certificate of Compliance endorsement application. (a... vessel meets § 154.1725(a) (4), (5), and (7). (9) If the vessel is a new gas vessel, or an existing...