Epithelial organization and cyst lumen expansion require efficient Sec13-Sec31-driven secretion.

PubMed

Townley, Anna K; Schmidt, Katy; Hodgson, Lorna; Stephens, David J

2012-02-01

Epithelial morphogenesis is directed by interactions with the underlying extracellular matrix. Secretion of collagen and other matrix components requires efficient coat complex II (COPII) vesicle formation at the endoplasmic reticulum. Here, we show that suppression of the outer layer COPII component, Sec13, in zebrafish embryos results in a disorganized gut epithelium. In human intestinal epithelial cells (Caco-2), Sec13 depletion causes defective epithelial polarity and organization on permeable supports. Defects are seen in the ability of cells to adhere to the substrate, form a monolayer and form intercellular junctions. When embedded in a three-dimensional matrix, Sec13-depleted Caco-2 cells form cysts but, unlike controls, are defective in lumen expansion. Incorporation of primary fibroblasts within the three-dimensional culture substantially restores normal morphogenesis. We conclude that efficient COPII-dependent secretion, notably assembly of Sec13-Sec31, is required to drive epithelial morphogenesis in both two- and three-dimensional cultures in vitro, as well as in vivo. Our results provide insight into the role of COPII in epithelial morphogenesis and have implications for the interpretation of epithelial polarity and organization assays in cell culture.

Epithelial organization and cyst lumen expansion require efficient Sec13–Sec31-driven secretion

PubMed Central

Townley, Anna K.; Schmidt, Katy; Hodgson, Lorna; Stephens, David J.

2012-01-01

Epithelial morphogenesis is directed by interactions with the underlying extracellular matrix. Secretion of collagen and other matrix components requires efficient coat complex II (COPII) vesicle formation at the endoplasmic reticulum. Here, we show that suppression of the outer layer COPII component, Sec13, in zebrafish embryos results in a disorganized gut epithelium. In human intestinal epithelial cells (Caco-2), Sec13 depletion causes defective epithelial polarity and organization on permeable supports. Defects are seen in the ability of cells to adhere to the substrate, form a monolayer and form intercellular junctions. When embedded in a three-dimensional matrix, Sec13-depleted Caco-2 cells form cysts but, unlike controls, are defective in lumen expansion. Incorporation of primary fibroblasts within the three-dimensional culture substantially restores normal morphogenesis. We conclude that efficient COPII-dependent secretion, notably assembly of Sec13–Sec31, is required to drive epithelial morphogenesis in both two- and three-dimensional cultures in vitro, as well as in vivo. Our results provide insight into the role of COPII in epithelial morphogenesis and have implications for the interpretation of epithelial polarity and organization assays in cell culture. PMID:22331354

A screen for over-secretion of proteins by yeast based on a dual component cellular phosphatase and immuno-chromogenic stain for exported bacterial alkaline phosphatase reporter

PubMed Central

2013-01-01

Background To isolate over-secretors, we subjected to saturation mutagenesis, a strain of P.pastoris exporting E. coli alkaline phosphatase (EAP) fused to the secretory domain of the yeast α factor pheromone through cellular PHO1/KEX2 secretory processing signals as the α-sec-EAP reporter protein. Direct chromogenic staining for α-sec-EAP activity is non-specific as its NBT/BCIP substrate cross-reacts with cellular phosphatases which can be inhibited with Levulinic acid. However, the parental E(P) strain only exports detectable levels of α-sec-EAP at 69 hours and not within the 36 hour period post-seeding required for effective screening with the consequent absence of a reference for secretion. We substituted the endogenous cellular phosphatase activity as a comparative reference for secretion rate and levels as well as for colony alignment while elevating specificity and sensitivity of detection of the exported protein with other innovative modifications of the immuno-chromogenic staining application for screening protein export mutants. Results Raising the specificity and utility of staining for α-sec-EAP activity required 5 modifications including some to published methods. These included, exploitation of endogenous phosphatase activity, reduction of the cell/protein burden, establishment of the direct relation between concentrations of transcriptional inducer and exported membrane immobilized protein and concentrations of protein exported into growth media, amplification of immuno-specificity and sensitivity of detection of α-sec-EAP reporter enzyme signal and restriction of staining to optimal concentrations of antisera and time periods. The resultant immuno-chromogenic screen allows for the detection of early secretion and as little as 1.3 fold over-secretion of α-sec-EAP reporter protein by E(M) mutants in the presence of 10 fold -216 fold higher concentrations of HSA. Conclusions The modified immuno-chromogenic screen is sensitive, specific and has led to the isolation of mutants E(M) over-secreting the α-sec-EAP reporter protein by a minimum of 50 fold higher levels than that exported by non-mutagenized E(P) parental strains. Unselected proteins were also over-secreted. PMID:23602005

Inhibitory effects of anthocyanins on secretion of Helicobacter pylori CagA and VacA toxins.

PubMed

Kim, Sa-Hyun; Park, Min; Woo, Hyunjun; Tharmalingam, Nagendran; Lee, Gyusang; Rhee, Ki-Jong; Eom, Yong Bin; Han, Sang Ik; Seo, Woo Duck; Kim, Jong Bae

2012-01-01

Anthocyanins have been studied as potential antimicrobial agents against Helicobacter pylori. We investigated whether the biosynthesis and secretion of cytotoxin-associated protein A (CagA) and vacuolating cytotoxin A (VacA) could be suppressed by anthocyanin treatment in vitro. H. pylori reference strain 60190 (CagA(+)/VacA(+)) was used in this study to investigate the inhibitory effects of anthocyanins; cyanidin 3-O-glucoside (C3G), peonidin 3-O-glucoside (Peo3G), pelargonidin 3-O-glucoside (Pel3G), and malvidin 3-O-glucoside (M3G) on expression and secretion of H. pylori toxins. Anthocyanins were added to bacterial cultures and Western blotting was used to determine secretion of CagA and VacA. Among them, we found that C3G inhibited secretion of CagA and VacA resulting in intracellular accumulation of CagA and VacA. C3G had no effect on cagA and vacA expression but suppressed secA transcription. As SecA is involved in translocation of bacterial proteins, the down-regulation of secA expression by C3G offers a mechanistic explanation for the inhibition of toxin secretion. To our knowledge, this is the first report suggesting that C3G inhibits secretion of the H. pylori toxins CagA and VacA via suppression of secA transcription.

Inhibitory Effects of Anthocyanins on Secretion of Helicobacter pylori CagA and VacA Toxins

PubMed Central

Kim, Sa-Hyun; Park, Min; Woo, Hyunjun; Tharmalingam, Nagendran; Lee, Gyusang; Rhee, Ki-Jong; Eom, Yong Bin; Han, Sang Ik; Seo, Woo Duck; Kim, Jong Bae

2012-01-01

Anthocyanins have been studied as potential antimicrobial agents against Helicobacter pylori. We investigated whether the biosynthesis and secretion of cytotoxin-associated protein A (CagA) and vacuolating cytotoxin A (VacA) could be suppressed by anthocyanin treatment in vitro. H. pylori reference strain 60190 (CagA+/VacA+) was used in this study to investigate the inhibitory effects of anthocyanins; cyanidin 3-O-glucoside (C3G), peonidin 3-O-glucoside (Peo3G), pelargonidin 3-O-glucoside (Pel3G), and malvidin 3-O-glucoside (M3G) on expression and secretion of H. pylori toxins. Anthocyanins were added to bacterial cultures and Western blotting was used to determine secretion of CagA and VacA. Among them, we found that C3G inhibited secretion of CagA and VacA resulting in intracellular accumulation of CagA and VacA. C3G had no effect on cagA and vacA expression but suppressed secA transcription. As SecA is involved in translocation of bacterial proteins, the down-regulation of secA expression by C3G offers a mechanistic explanation for the inhibition of toxin secretion. To our knowledge, this is the first report suggesting that C3G inhibits secretion of the H. pylori toxins CagA and VacA via suppression of secA transcription. PMID:23155357

32 CFR Attachment D to Subpart B... - Decision Tables

Code of Federal Regulations, 2011 CFR

2011-07-01

... date NACLC or SSBI Conf, Sec; “L” Top Secret, SCI; “Q” None None SSBI B None; Conf, Sec; “L” Current or out of date NACLC Out of date SSBI TS, SCI; “Q” SSBI-PR C Table 2—Reinvestigation Requirements If the... NACLC Secret; “L” 0 to 9 yrs 11 mos None (note 1) 10 yrs. or more NACLC Top Secret, SCI; “Q” 0 to 4 yrs...

Ubiquitin-specific protease 8 deubiquitinates Sec31A and decreases large COPII carriers and collagen IV secretion.

PubMed

Kawaguchi, Kohei; Endo, Akinori; Fukushima, Toshiaki; Madoka, Yuka; Tanaka, Toshiaki; Komada, Masayuki

2018-05-15

Nascent cargo proteins in the endoplasmic reticulum are transported to the Golgi by COPII carriers. Typical COPII vesicles are 60-70 nm in diameter, and much larger macromolecules, such as procollagen, are transported by atypical large COPII carriers in mammalian cells. The formation of large COPII carriers is enhanced by Cul3 ubiquitin ligase, which mono-ubiquitinates Sec31A, a COPII coat protein. However, the deubiquitinating enzyme for Sec31A was unclear. Here, we show that the deubiquitinating enzyme USP8 interacts with and deubiquitinates Sec31A. The interaction was mediated by the adaptor protein STAM1. USP8 overexpression inhibited the formation of large COPII carriers. By contrast, USP8 knockdown caused the accumulation of COPII coat proteins around the cis-Golgi, promoted the intracellular trafficking of procollagen IV from the endoplasmic reticulum to the Golgi, and increased collagen IV secretion. We concluded that USP8 deubiquitinates Sec31A and inhibits the formation of large COPII carriers, thereby suppressing collagen IV secretion. Copyright © 2018 Elsevier Inc. All rights reserved.

The genome sequence of the facultative intracellular pathogen Brucella melitensis.

PubMed

DelVecchio, Vito G; Kapatral, Vinayak; Redkar, Rajendra J; Patra, Guy; Mujer, Cesar; Los, Tamara; Ivanova, Natalia; Anderson, Iain; Bhattacharyya, Anamitra; Lykidis, Athanasios; Reznik, Gary; Jablonski, Lynn; Larsen, Niels; D'Souza, Mark; Bernal, Axel; Mazur, Mikhail; Goltsman, Eugene; Selkov, Eugene; Elzer, Philip H; Hagius, Sue; O'Callaghan, David; Letesson, Jean-Jacques; Haselkorn, Robert; Kyrpides, Nikos; Overbeek, Ross

2002-01-08

Brucella melitensis is a facultative intracellular bacterial pathogen that causes abortion in goats and sheep and Malta fever in humans. The genome of B. melitensis strain 16M was sequenced and found to contain 3,294,935 bp distributed over two circular chromosomes of 2,117,144 bp and 1,177,787 bp encoding 3,197 ORFs. By using the bioinformatics suite ERGO, 2,487 (78%) ORFs were assigned functions. The origins of replication of the two chromosomes are similar to those of other alpha-proteobacteria. Housekeeping genes, including those involved in DNA replication, transcription, translation, core metabolism, and cell wall biosynthesis, are distributed on both chromosomes. Type I, II, and III secretion systems are absent, but genes encoding sec-dependent, sec-independent, and flagella-specific type III, type IV, and type V secretion systems as well as adhesins, invasins, and hemolysins were identified. Several features of the B. melitensis genome are similar to those of the symbiotic Sinorhizobium meliloti.

The genome sequence of the facultative intracellular pathogen Brucella melitensis

PubMed Central

DelVecchio, Vito G.; Kapatral, Vinayak; Redkar, Rajendra J.; Patra, Guy; Mujer, Cesar; Los, Tamara; Ivanova, Natalia; Anderson, Iain; Bhattacharyya, Anamitra; Lykidis, Athanasios; Reznik, Gary; Jablonski, Lynn; Larsen, Niels; D'Souza, Mark; Bernal, Axel; Mazur, Mikhail; Goltsman, Eugene; Selkov, Eugene; Elzer, Philip H.; Hagius, Sue; O'Callaghan, David; Letesson, Jean-Jacques; Haselkorn, Robert; Kyrpides, Nikos; Overbeek, Ross

2002-01-01

Brucella melitensis is a facultative intracellular bacterial pathogen that causes abortion in goats and sheep and Malta fever in humans. The genome of B. melitensis strain 16M was sequenced and found to contain 3,294,935 bp distributed over two circular chromosomes of 2,117,144 bp and 1,177,787 bp encoding 3,197 ORFs. By using the bioinformatics suite ERGO, 2,487 (78%) ORFs were assigned functions. The origins of replication of the two chromosomes are similar to those of other α-proteobacteria. Housekeeping genes, including those involved in DNA replication, transcription, translation, core metabolism, and cell wall biosynthesis, are distributed on both chromosomes. Type I, II, and III secretion systems are absent, but genes encoding sec-dependent, sec-independent, and flagella-specific type III, type IV, and type V secretion systems as well as adhesins, invasins, and hemolysins were identified. Several features of the B. melitensis genome are similar to those of the symbiotic Sinorhizobium meliloti. PMID:11756688

Structural Similarities and Differences between Two Functionally Distinct SecA Proteins, Mycobacterium tuberculosis SecA1 and SecA2

PubMed Central

Swanson, Stephanie; Ioerger, Thomas R.; Rigel, Nathan W.; Miller, Brittany K.; Braunstein, Miriam

2015-01-01

ABSTRACT While SecA is the ATPase component of the major bacterial secretory (Sec) system, mycobacteria and some Gram-positive pathogens have a second paralog, SecA2. In bacteria with two SecA paralogs, each SecA is functionally distinct, and they cannot compensate for one another. Compared to SecA1, SecA2 exports a distinct and smaller set of substrates, some of which have roles in virulence. In the mycobacterial system, some SecA2-dependent substrates lack a signal peptide, while others contain a signal peptide but possess features in the mature protein that necessitate a role for SecA2 in their export. It is unclear how SecA2 functions in protein export, and one open question is whether SecA2 works with the canonical SecYEG channel to export proteins. In this study, we report the structure of Mycobacterium tuberculosis SecA2 (MtbSecA2), which is the first structure of any SecA2 protein. A high level of structural similarity is observed between SecA2 and SecA1. The major structural difference is the absence of the helical wing domain, which is likely to play a role in how MtbSecA2 recognizes its unique substrates. Importantly, structural features critical to the interaction between SecA1 and SecYEG are preserved in SecA2. Furthermore, suppressor mutations of a dominant-negative secA2 mutant map to the surface of SecA2 and help identify functional regions of SecA2 that may promote interactions with SecYEG or the translocating polypeptide substrate. These results support a model in which the mycobacterial SecA2 works with SecYEG. IMPORTANCE SecA2 is a paralog of SecA1, which is the ATPase of the canonical bacterial Sec secretion system. SecA2 has a nonredundant function with SecA1, and SecA2 exports a distinct and smaller set of substrates than SecA1. This work reports the crystal structure of SecA2 of Mycobacterium tuberculosis (the first SecA2 structure reported for any organism). Many of the structural features of SecA1 are conserved in the SecA2 structure, including putative contacts with the SecYEG channel. Several structural differences are also identified that could relate to the unique function and selectivity of SecA2. Suppressor mutations of a secA2 mutant map to the surface of SecA2 and help identify functional regions of SecA2 that may promote interactions with SecYEG. PMID:26668263

Hsp90/Sec22b promotes unconventional secretion of mature-IL-1β through an autophagosomal carrier in porcine alveolar macrophages during Mycoplasma hyopneumoniae infection.

PubMed

Zhang, Zhenzhen; Wei, Yanna; Liu, Beibei; Wu, Yuzi; Wang, Haiyan; Xie, Xing; Feng, Zhixin; Shao, Guoqing; Xiong, Qiyan

2018-06-20

Interleukin-1β (IL-1β) is a critical inflammatory regulator in response to Mycoplasma hyopneumoniae infection. However, the mechanism involved in the secretion of IL-1β during Mycoplasma hyopneumoniae infection is unclear. In this study, we demonstrated that Mycoplasma hyopneumoniae infection increased the secretion of mature-IL-1β (m-IL-1β), but not pro-IL-1β, in porcine alveolar macrophages. Moreover, Mycoplasma hyopneumoniae infection promoted the generation of autophagosomes, which attributed to the unconventional secretion of m-IL-1β. Further results revealed that Hsp90 was required for the entry of m-IL-1β into autophagosomes during Mycoplasma hyopneumoniae infection. The fusion of m-IL-1β-containing autophagosome and plasma membranes was regulated by Sec22b and independent of lysosomal dysfunction. In conclusion, we provide evidence that Hsp90/Sec22b promotes the unconventional secretion of IL-1β through an autophagosomal carrier during Mycoplasma hyopneumoniae infection. The elucidation of the molecular and cellular machinery in Mycoplasma hyopneumoniae infected mammalian cells in this study suggests avenues for further study and applications and paves the way for novel therapeutic strategies to prevent tissue damage in mycoplasma-associated diseases. Copyright © 2018. Published by Elsevier Ltd.

Yeast syntaxins Sso1p and Sso2p belong to a family of related membrane proteins that function in vesicular transport.

PubMed Central

Aalto, M K; Ronne, H; Keränen, S

1993-01-01

The yeast SEC1 gene encodes a hydrophilic protein that functions at the terminal stage in secretion. We have cloned two yeast genes, SSO1 and SSO2, which in high copy number can suppress sec1 mutations and also mutations in several other late acting SEC genes, such as SEC3, SEC5, SEC9 and SEC15. SSO1 and SSO2 encode small proteins with N-terminal hydrophilic domains and C-terminal hydrophobic tails. The two proteins are 72% identical in sequence and together perform an essential function late in secretion. Sso1p and Sso2p show significant sequence similarity to six other proteins. Two of these, Sed5p and Pep12p, are yeast proteins that function in transport from ER to Golgi and from Golgi to the vacuole, respectively. Also related to Sso1p and Sso2p are three mammalian proteins: epimorphin, syntaxin A/HPC-1 and syntaxin B. A nematode cDNA product also belongs to the new protein family. The new protein family is thus present in a wide variety of eukaryotic cells, where its members function at different stages in vesicular transport. Images PMID:8223426

Study of the plant COPII vesicle coat subunits by functional complementation of yeast Saccharomyces cerevisiae mutants.

PubMed

De Craene, Johan-Owen; Courte, Fanny; Rinaldi, Bruno; Fitterer, Chantal; Herranz, Mari Carmen; Schmitt-Keichinger, Corinne; Ritzenthaler, Christophe; Friant, Sylvie

2014-01-01

The formation and budding of endoplasmic reticulum ER-derived vesicles depends on the COPII coat protein complex that was first identified in yeast Saccharomyces cerevisiae. The ER-associated Sec12 and the Sar1 GTPase initiate the COPII coat formation by recruiting the Sec23-Sec24 heterodimer following the subsequent recruitment of the Sec13-Sec31 heterotetramer. In yeast, there is usually one gene encoding each COPII protein and these proteins are essential for yeast viability, whereas the plant genome encodes multiple isoforms of all COPII subunits. Here, we used a systematic yeast complementation assay to assess the functionality of Arabidopsis thaliana COPII proteins. In this study, the different plant COPII subunits were expressed in their corresponding temperature-sensitive yeast mutant strain to complement their thermosensitivity and secretion phenotypes. Secretion was assessed using two different yeast cargos: the soluble α-factor pheromone and the membranous v-SNARE (vesicle-soluble NSF (N-ethylmaleimide-sensitive factor) attachment protein receptor) Snc1 involved in the fusion of the secretory vesicles with the plasma membrane. This complementation study allowed the identification of functional A. thaliana COPII proteins for the Sec12, Sar1, Sec24 and Sec13 subunits that could represent an active COPII complex in plant cells. Moreover, we found that AtSec12 and AtSec23 were co-immunoprecipitated with AtSar1 in total cell extract of 15 day-old seedlings of A. thaliana. This demonstrates that AtSar1, AtSec12 and AtSec23 can form a protein complex that might represent an active COPII complex in plant cells.

Somatic overgrowth associated with homozygous mutations in both MAN1B1 and SEC23A

PubMed Central

Gupta, Swati; Fahiminiya, Somayyeh; Wang, Tracy; Dempsey Nunez, Laura; Rosenblatt, David S.; Gibson, William T.; Gilfix, Brian; Bergeron, John J. M.; Jerome-Majewska, Loydie A.

2016-01-01

Using whole-exome sequencing, we identified homozygous mutations in two unlinked genes, SEC23A c.1200G>C (p.M400I) and MAN1B1 c.1000C>T (p.R334C), associated with congenital birth defects in two patients from a consanguineous family. Patients presented with carbohydrate-deficient transferrin, tall stature, obesity, macrocephaly, and maloccluded teeth. The parents were healthy heterozygous carriers for both mutations and an unaffected sibling with tall stature carried the heterozygous mutation in SEC23A only. Mutations in SEC23A are responsible for craniolenticosultura dysplasia (CLSD). CLSD patients are short, have late-closing fontanels, and have reduced procollagen (pro-COL1A1) secretion because of abnormal pro-COL1A1 retention in the endoplasmic reticulum (ER). The mutation we identified in MAN1B1 was previously associated with reduced MAN1B1 protein and congenital disorders of glycosylation (CDG). CDG patients are also short, are obese, and have abnormal glycan remodeling. Molecular analysis of fibroblasts from the family revealed normal levels of SEC23A in all cells and reduced levels of MAN1B1 in cells with heterozygous or homozygous mutations in SEC23A and MAN1B1. Secretion of pro-COL1A1 was increased in fibroblasts from the siblings and patients, and pro-COL1A1 was retained in Golgi of heterozygous and homozygous mutant cells, although intracellular pro-COL1A1 was increased in patient fibroblasts only. We postulate that increased pro-COL1A1 secretion is responsible for tall stature in these patients and an unaffected sibling, and not previously discovered in patients with mutations in either SEC23A or MAN1B1. The patients in this study share biochemical and cellular characteristics consistent with mutations in MAN1B1 and SEC23A, indicating a digenic disease. PMID:27148587

Unraveling novel broad-spectrum antibacterial targets in food and waterborne pathogens using comparative genomics and protein interaction network analysis.

PubMed

Jadhav, Ankush; Shanmugham, Buvaneswari; Rajendiran, Anjana; Pan, Archana

2014-10-01

Food and waterborne diseases are a growing concern in terms of human morbidity and mortality worldwide, even in the 21st century, emphasizing the need for new therapeutic interventions for these diseases. The current study aims at prioritizing broad-spectrum antibacterial targets, present in multiple food and waterborne bacterial pathogens, through a comparative genomics strategy coupled with a protein interaction network analysis. The pathways unique and common to all the pathogens under study (viz., methane metabolism, d-alanine metabolism, peptidoglycan biosynthesis, bacterial secretion system, two-component system, C5-branched dibasic acid metabolism), identified by comparative metabolic pathway analysis, were considered for the analysis. The proteins/enzymes involved in these pathways were prioritized following host non-homology analysis, essentiality analysis, gut flora non-homology analysis and protein interaction network analysis. The analyses revealed a set of promising broad-spectrum antibacterial targets, present in multiple food and waterborne pathogens, which are essential for bacterial survival, non-homologous to host and gut flora, and functionally important in the metabolic network. The identified broad-spectrum candidates, namely, integral membrane protein/virulence factor (MviN), preprotein translocase subunits SecB and SecG, carbon storage regulator (CsrA), and nitrogen regulatory protein P-II 1 (GlnB), contributed by the peptidoglycan pathway, bacterial secretion systems and two-component systems, were also found to be present in a wide range of other disease-causing bacteria. Cytoplasmic proteins SecG, CsrA and GlnB were considered as drug targets, while membrane proteins MviN and SecB were classified as vaccine targets. The identified broad-spectrum targets can aid in the design and development of antibacterial agents not only against food and waterborne pathogens but also against other pathogens. Copyright © 2014 Elsevier B.V. All rights reserved.

The Three Streptomyces lividans HtrA-Like Proteases Involved in the Secretion Stress Response Act in a Cooperative Manner

PubMed Central

Vicente, Rebeca L.; Gullón, Sonia; Marín, Silvia; Mellado, Rafael P.

2016-01-01

Overproduction of Sec-proteins in S. lividans accumulates misfolded proteins outside of the cytoplasmic membrane where the accumulated proteins interfere with the correct functioning of the secretion machinery and with the correct cell functionality, triggering the expression in S. lividans of a CssRS two-component system which regulates the degradation of the accumulated protein, the so-called secretion stress response. Optimization of secretory protein production via the Sec route requires the identification and characterisation of quality factors involved in this process. The phosphorylated regulator (CssR) interacts with the regulatory regions of three genes encoding three different HtrA-like proteases. Individual mutations in each of these genes render degradation of the misfolded protein inoperative, and propagation in high copy number of any of the three proteases encoding genes results on indiscriminate alpha-amylase degradation. None of the proteases could complement the other two deficiencies and only propagation of each single copy protease gene can restore its own deficiency. The obtained results strongly suggest that the synthesis of the three HtrA-like proteases needs to be properly balanced to ensure the effective degradation of misfolded overproduced secretory proteins and, at the same time, avoid negative effects in the secreted proteins and the secretion machinery. This is particularly relevant when considering the optimisation of Streptomyces strains for the overproduction of homologous or heterologous secretory proteins of industrial application. PMID:27977736

Regulation of exocytosis by the exocyst subunit Sec6 and the SM protein Sec1.

PubMed

Morgera, Francesca; Sallah, Margaret R; Dubuke, Michelle L; Gandhi, Pallavi; Brewer, Daniel N; Carr, Chavela M; Munson, Mary

2012-01-01

Trafficking of protein and lipid cargo through the secretory pathway in eukaryotic cells is mediated by membrane-bound vesicles. Secretory vesicle targeting and fusion require a conserved multisubunit protein complex termed the exocyst, which has been implicated in specific tethering of vesicles to sites of polarized exocytosis. The exocyst is directly involved in regulating soluble N-ethylmaleimide-sensitive factor (NSF) attachment protein receptor (SNARE) complexes and membrane fusion through interactions between the Sec6 subunit and the plasma membrane SNARE protein Sec9. Here we show another facet of Sec6 function-it directly binds Sec1, another SNARE regulator, but of the Sec1/Munc18 family. The Sec6-Sec1 interaction is exclusive of Sec6-Sec9 but compatible with Sec6-exocyst assembly. In contrast, the Sec6-exocyst interaction is incompatible with Sec6-Sec9. Therefore, upon vesicle arrival, Sec6 is proposed to release Sec9 in favor of Sec6-exocyst assembly and to simultaneously recruit Sec1 to sites of secretion for coordinated SNARE complex formation and membrane fusion.

Extracellular secretion of pectate lyase by the Erwinia chrysanthemi out pathway is dependent upon Sec-mediated export across the inner membrane.

PubMed Central

He, S Y; Schoedel, C; Chatterjee, A K; Collmer, A

1991-01-01

The plant pathogenic enterobacterium Erwinia chrysanthemi EC16 secretes several extracellular, plant cell wall-degrading enzymes, including pectate lyase isozyme PelE. Secretion kinetics of 35S-labeled PelE indicated that the precursor of PelE was rapidly processed by the removal of the amino-terminal signal peptide and that the resulting mature PelE remained cell bound for less than 60 s before being secreted to the bacterial medium. PelE-PhoA (alkaline phosphatase) hybrid proteins generated in vivo by TnphoA insertions were mostly localized in the periplasm of E. chrysanthemi, and one hybrid protein was observed to be associated with the outer membrane of E. chrysanthemi in an out gene-dependent manner. A gene fusion resulting in the substitution of the beta-lactamase signal peptide for the first six amino acids of the PelE signal peptide did not prevent processing or secretion of PelE in E. chrysanthemi. When pelE was overexpressed, mature PelE protein accumulated in the periplasm rather than the cytoplasm in cells of E. chrysanthemi and Escherichia coli MC4100 (pCPP2006), which harbors a functional cluster of E. chrysanthemi out genes. Removal of the signal peptide from pre-PelE was SecA dependent in E. coli MM52 even in the presence of the out gene cluster. These data indicate that the extracellular secretion of pectic enzymes by E. chrysanthemi is an extension of the Sec-dependent pathway for general export of proteins across the bacterial inner membrane. Images PMID:1829728

Role of the Carboxy Terminus of SecA in Iron Acquisition, Protein Translocation, and Virulence of the Bacterial Pathogen Acinetobacter baumannii

PubMed Central

Fiester, Steven E.; Nwugo, Chika C.; Penwell, William F.; Neary, John M.; Beckett, Amber C.; Arivett, Brock A.; Schmidt, Robert E.; Geiger, Sarah C.; Connerly, Pamela L.; Menke, Sharon M.; Tomaras, Andrew P.

2015-01-01

Acinetobacter baumannii is a Gram-negative opportunistic nosocomial pathogen that causes pneumonia and soft tissue and systemic infections. Screening of a transposon insertion library of A. baumannii ATCC 19606T resulted in the identification of the 2010 derivative, which, although capable of growing well in iron-rich media, failed to prosper under iron chelation. Genetic, molecular, and functional assays showed that 2010's iron utilization-deficient phenotype is due to an insertion within the 3′ end of secA, which results in the production of a C-terminally truncated derivative of SecA. SecA plays a critical role in protein translocation through the SecYEG membrane channel. Accordingly, the secA mutation resulted in undetectable amounts of the ferric acinetobactin outer membrane receptor protein BauA while not affecting the production of other acinetobactin membrane protein transport components, such as BauB and BauE, or the secretion of acinetobactin by 2010 cells cultured in the presence of subinhibitory concentrations of the synthetic iron chelator 2,2′-dipyridyl. Outer membrane proteins involved in nutrient transport, adherence, and biofilm formation were also reduced in 2010. The SecA truncation also increased production of 30 different proteins, including proteins involved in adaptation/tolerance responses. Although some of these protein changes could negatively affect the pathobiology of the 2010 derivative, its virulence defect is mainly due to its inability to acquire iron via the acinetobactin-mediated system. These results together indicate that although the C terminus of the A. baumannii ATCC 19606T SecA is not essential for viability, it plays a critical role in the production and translocation of different proteins and virulence. PMID:25605767

Haloarchaeal Protein Translocation via the Twin Arginine Translocation Pathway

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

Pohlschroder Mechthild

2009-02-03

Protein transport across hydrophobic membranes that partition cellular compartments is essential in all cells. The twin arginine translocation (Tat) pathway transports proteins across the prokaryotic cytoplasmic membranes. Distinct from the universally conserved Sec pathway, which secretes unfolded proteins, the Tat machinery is unique in that it secretes proteins in a folded conformation, making it an attractive pathway for the transport and secretion of heterologously expressed proteins that are Sec-incompatible. During the past 7 years, the DOE-supported project has focused on the characterization of the diversity of bacterial and archaeal Tat substrates as well as on the characterization of the Tatmore » pathway of a model archaeon, Haloferax volcanii, a member of the haloarchaea. We have demonstrated that H. volcanii uses this pathway to transport most of its secretome.« less

Cellular and molecular mechanism for secretory autophagy.

PubMed

Kimura, Tomonori; Jia, Jingyue; Claude-Taupin, Aurore; Kumar, Suresh; Choi, Seong Won; Gu, Yuexi; Mudd, Michal; Dupont, Nicolas; Jiang, Shanya; Peters, Ryan; Farzam, Farzin; Jain, Ashish; Lidke, Keith A; Adams, Christopher M; Johansen, Terje; Deretic, Vojo

2017-06-03

Macroautophagy/autophagy plays a role in unconventional secretion of leaderless cytosolic proteins. Whether and how secretory autophagy diverges from conventional degradative autophagy is unclear. We have shown that the prototypical secretory autophagy cargo IL1B/IL-1β (interleukin 1 β) is recognized by TRIM16, and that this first to be identified secretory autophagy receptor interacts with the R-SNARE SEC22B to jointly deliver cargo to the MAP1LC3B-II-positive sequestration membranes. Cargo secretion is unaffected by knockdowns of STX17, a SNARE catalyzing autophagosome-lysosome fusion as a prelude to cargo degradation. Instead, SEC22B in combination with plasma membrane syntaxins completes cargo secretion. Thus, secretory autophagy diverges from degradative autophagy by using specialized receptors and a dedicated SNARE machinery to bypass fusion with lysosomes.

Phytophthora infestans RXLR Effector AVR1 Interacts with Exocyst Component Sec5 to Manipulate Plant Immunity.

PubMed

Du, Yu; Mpina, Mohamed H; Birch, Paul R J; Bouwmeester, Klaas; Govers, Francine

2015-11-01

Phytophthora infestans secretes numerous RXLR effectors that modulate host defense and thereby pave the way for successful invasion. Here, we show that the RXLR effector AVR1 is a virulence factor that promotes colonization and suppresses callose deposition, a hallmark of basal defense. To identify host targets of AVR1, we performed yeast two-hybrid screens and selected Sec5 as a candidate. Sec5 is a subunit of the exocyst, a protein complex that is involved in vesicle trafficking. AVR1-like (A-L), a close homolog of AVR1, also acts as a virulence factor, but unlike AVR1, A-L does not suppress CRINKLER2 (CRN2)-induced cell death or interact with Sec5. Compared with AVR1, A-L is shorter and lacks the carboxyl-terminal tail, the T-region that is crucial for CRN2-induced cell death suppression and Sec5 interaction. In planta analyses revealed that AVR1 and Sec5 are in close proximity, and coimmunoprecipitation confirmed the interaction. Sec5 is required for secretion of the pathogenesis-related protein PR-1 and callose deposition and also plays a role in CRN2-induced cell death. Our findings show that P. infestans manipulates an exocyst subunit and thereby potentially disturbs vesicle trafficking, a cellular process that is important for basal defense. This is a novel strategy that oomycete pathogens exploit to modulate host defense. © 2015 American Society of Plant Biologists. All Rights Reserved.

Phytophthora infestans RXLR Effector AVR1 Interacts with Exocyst Component Sec5 to Manipulate Plant Immunity1[OPEN

PubMed Central

Du, Yu; Mpina, Mohamed H.; Birch, Paul R.J.; Bouwmeester, Klaas; Govers, Francine

2015-01-01

Phytophthora infestans secretes numerous RXLR effectors that modulate host defense and thereby pave the way for successful invasion. Here, we show that the RXLR effector AVR1 is a virulence factor that promotes colonization and suppresses callose deposition, a hallmark of basal defense. To identify host targets of AVR1, we performed yeast two-hybrid screens and selected Sec5 as a candidate. Sec5 is a subunit of the exocyst, a protein complex that is involved in vesicle trafficking. AVR1-like (A-L), a close homolog of AVR1, also acts as a virulence factor, but unlike AVR1, A-L does not suppress CRINKLER2 (CRN2)-induced cell death or interact with Sec5. Compared with AVR1, A-L is shorter and lacks the carboxyl-terminal tail, the T-region that is crucial for CRN2-induced cell death suppression and Sec5 interaction. In planta analyses revealed that AVR1 and Sec5 are in close proximity, and coimmunoprecipitation confirmed the interaction. Sec5 is required for secretion of the pathogenesis-related protein PR-1 and callose deposition and also plays a role in CRN2-induced cell death. Our findings show that P. infestans manipulates an exocyst subunit and thereby potentially disturbs vesicle trafficking, a cellular process that is important for basal defense. This is a novel strategy that oomycete pathogens exploit to modulate host defense. PMID:26336092

Genome Sequence and Analysis of the Soil Cellulolytic ActinomyceteThermobifida fusca

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

Lykidis, Athanasios; Mavromatis, Konstantinos; Ivanova, Natalia

Thermobifida fusca is a moderately thermophilic soilbacterium that belongs to Actinobacteria. 3 It is a major degrader ofplant cell walls and has been used as a model organism for the study of 4secreted, thermostable cellulases. The complete genome sequence showedthat T. fusca has a 5 single circular chromosome of 3642249 bp predictedto encode 3117 proteins and 65 RNA6 species with a coding densityof 85percent. Genome analysis revealed the existence of 29 putative 7glycoside hydrolases in addition to the previously identified cellulasesand xylanases. The 8 glycosyl hydrolases include enzymes predicted toexhibit mainly dextran/starch and xylan 9 degrading functions. T. fuscapossesses twomore » protein secretion systems: the sec general secretion 10system and the twin-arginine translocation system. Several of thesecreted cellulases have 11 sequence signatures indicating theirsecretion may be mediated by the twin-arginine12 translocation system. T.fusca has extensive transport systems for import of carbohydrates 13coupled to transcriptional regulators controlling the expression of thetransporters and14 glycosylhydrolases. In addition to providing anoverview of the physiology of a soil 15 actinomycete, this study presentsinsights on the transcriptional regulation and secretion of16 cellulaseswhich may facilitate the industrial exploitation of thesesystems.« less

Two-way communication between SecY and SecA suggests a Brownian ratchet mechanism for protein translocation.

PubMed

Allen, William John; Corey, Robin Adam; Oatley, Peter; Sessions, Richard Barry; Baldwin, Steve A; Radford, Sheena E; Tuma, Roman; Collinson, Ian

2016-05-16

The essential process of protein secretion is achieved by the ubiquitous Sec machinery. In prokaryotes, the drive for translocation comes from ATP hydrolysis by the cytosolic motor-protein SecA, in concert with the proton motive force (PMF). However, the mechanism through which ATP hydrolysis by SecA is coupled to directional movement through SecYEG is unclear. Here, we combine all-atom molecular dynamics (MD) simulations with single molecule FRET and biochemical assays. We show that ATP binding by SecA causes opening of the SecY-channel at long range, while substrates at the SecY-channel entrance feed back to regulate nucleotide exchange by SecA. This two-way communication suggests a new, unifying 'Brownian ratchet' mechanism, whereby ATP binding and hydrolysis bias the direction of polypeptide diffusion. The model represents a solution to the problem of transporting inherently variable substrates such as polypeptides, and may underlie mechanisms of other motors that translocate proteins and nucleic acids.

A fluorescent reporter protein containing AtRMR1 domains is targeted to the storage and central vacuoles in Arabidopsis thaliana and tobacco leaf cells.

PubMed

Scabone, Camila María; Frigerio, Lorenzo; Petruccelli, Silvana

2011-10-01

To develop a new strategy to target recombinant proteins to the vacuolar storage system in transgenic plants, the ability of the transmembrane and cytosolic domains of Arabidopsis receptor homology-transmembrane-RING H2-1 (AtRMR1) was evaluated. A secreted version of RFP (secRFP) and a fusion of it to the transmembrane and cytosolic domains of AtRMR1 (RFP-TMCT) were produced and studied both in transient and stable expression assays. Transient expression in leaves of Nicotiana tabacum showed that secRFP is secreted to the apoplast while its fusion to TMCT of AtRMR1 is sufficient to prevent secretion of the reporter. In tobacco leaves, RFP-TMCT reporter showed an endoplasmic reticulum pattern in early expression stages while in late expression stages, it was found in the vacuolar lumen. For the first time, the role of TM and CT domains of AtRMR1 in stable expression in Arabidopsis thaliana is presented; the fusion of TMCT to secRFP is sufficient to sort RFP to the lumen of the central vacuoles in leaves and roots and to the lumen of PSV in cotyledons of mature embryos. In addition, biochemical studies performed in extract from transgenic plants showed that RFP-TMCT is an integral membrane protein. Full-length RFP-TMCT was also found in the vacuolar lumen, suggesting internalization into destination vacuole. Not colocalization of RFP-TMCT with tonoplast and plasma membrane markers were observed. This membrane vacuolar determinant sorting signal could be used for future application in molecular pharming as an alternative means to sort proteins of interest to vacuoles.

A Phytase-Based Reporter System for Identification of Functional Secretion Signals in Bifidobacteria

PubMed Central

Osswald, Annika; Westermann, Christina; Sun, Zhongke; Riedel, Christian U.

2015-01-01

Health-promoting effects have been attributed to a number of Bifidobacterium sp. strains. These effects as well as the ability to colonise the host depend on secreted proteins. Moreover, rational design of protein secretion systems bears the potential for the generation of novel probiotic bifidobacteria with improved health-promoting or therapeutic properties. To date, there is only very limited data on secretion signals of bifidobacteria available. Using in silico analysis, we demonstrate that all bifidobacteria encode the major components of Sec-dependent secretion machineries but only B. longum strains harbour Tat protein translocation systems. A reporter plasmid for secretion signals in bifidobacteria was established by fusing the coding sequence of the signal peptide of a sialidase of Bifidobacterium bifidum S17 to the phytase gene appA of E. coli. The recombinant strain showed increased phytase activity in spent culture supernatants and reduced phytase levels in crude extracts compared to the control indicating efficient phytase secretion. The reporter plasmid was used to screen seven predicted signal peptides in B. bifidum S17 and B. longum E18. The tested signal peptides differed substantially in their efficacy to mediate protein secretion in different host strains. An efficient signal peptide was used for expression and secretion of a therapeutically relevant protein in B. bifidum S17. Expression of a secreted cytosine deaminase led to a 100-fold reduced sensitivity of B. bifidum S17 to 5-fluorocytosine compared to the non-secreted cytosine deaminase suggesting efficient conversion of 5-fluorocytosine to the cytotoxic cancer drug 5-fluorouracil by cytosine deaminase occurred outside the bacterial cell. Selection of appropriate signal peptides for defined protein secretion might improve therapeutic efficacy as well as probiotic properties of bifidobacteria. PMID:26086721

Secretion of TcpF by the Vibrio cholerae Toxin-Coregulated Pilus Biogenesis Apparatus Requires an N-Terminal Determinant

PubMed Central

Megli, Christina J.

2013-01-01

Type IV pili are important for microcolony formation, biofilm formation, twitching motility, and attachment. We and others have shown that type IV pili are important for protein secretion across the outer membrane, similar to type II secretion systems. This study explored the relationship between protein secretion and pilus formation in Vibrio cholerae. The toxin-coregulated pilus (TCP), a type IV pilus required for V. cholerae pathogenesis, is necessary for the secretion of the colonization factor TcpF (T. J. Kirn, N. Bose, and R. K. Taylor, Mol. Microbiol. 49:81–92, 2003). This phenomenon is not unique to V. cholerae; secreted virulence factors that are dependent on the presence of components of the type IV pilus biogenesis apparatus for secretion have been reported with Dichelobacter nodosus (R. M. Kennan, O. P. Dhungyel, R. J. Whittington, J. R. Egerton, and J. I. Rood, J. Bacteriol. 183:4451–4458, 2001) and Francisella tularensis (A. J. Hager et al., Mol. Microbiol. 62:227–237, 2006). Using site-directed mutagenesis, we demonstrated that the secretion of TcpF is dependent on the presence of selected amino acid R groups at position five. We were unable to find other secretion determinants, suggesting that Y5 is the major secretion determinant within TcpF. We also report that proteins secreted in a type IV pilus biogenesis apparatus-dependent manner have a YXS motif within the first 15 amino acids following the Sec cleavage site. The YXS motif is not present in proteins secreted by type II secretion systems, indicating that this is unique to type IV pilus-mediated secretion. Moreover, we show that TcpF interacts with the pilin TcpA, suggesting that these proteins are secreted by the type IV pilus biogenesis system. These data provide a starting point for understanding how type IV pili can mediate secretion of virulence factors important for bacterial pathogenesis. PMID:23564177

Inhibition of rabbit platelet activation in vitro by antagonists of platelet-activating factor (PAF)

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

Cox, C.P.; Wood, K.L.

1986-03-05

The authors used washed, (/sup 3/H)serotonin-labeled rabbit platelets to study the in vitro aggregation and secretion responses induced by graded doses of PAF in the presence or absence of specific antagonists of PAF. These antagonists included CV-3988, L-652,731, triazolam and alprazolam. Platelets were pretreated with either an antagonist or the appropriate diluent for 60 sec prior to the addition of PAF (2 x 10/sup -10/ to 2 x 10/sup -7/ M). Aggregation was monitored continuously and recorded as the height of the aggregation tracing at 60 sec post-PAF. Secretion of (/sup 3/H)-serotonin was measured in a sample of the plateletsmore » removed at 60 sec post-PAF. When 2 x 10/sup -10/ M PAF was used as the stimulus, the concentration of antagonist needed for 50% inhibition (IC/sub 50/) of secretion was obtained at 0.05 ..mu..M, 0.15 ..mu..M, 0.6 ..mu..M and 2.5 ..mu..M, respectively, for L-652,731, CV-3988, triazolam and alprazolam. The corresponding IC/sub 50/ for aggregation was obtained at 0.2 ..mu..M, 0.1 ..mu..M, 1.5 ..mu..M and 6.5 ..mu..M, respectively. The inhibitory effects of these antagonists could be overcome by increasing the dose of PAF used. Although all of the antagonists were capable of completely inhibiting platelet aggregation and secretion, L-652,731 was the most potent PAF antagonist on a molar basis.« less

Bacillus anthracis secretome time course under host-simulated conditions and identification of immunogenic proteins.

PubMed

Walz, Alexander; Mujer, Cesar V; Connolly, Joseph P; Alefantis, Tim; Chafin, Ryan; Dake, Clarissa; Whittington, Jessica; Kumar, Srikanta P; Khan, Akbar S; DelVecchio, Vito G

2007-07-27

The secretion time course of Bacillus anthracis strain RA3R (pXO1+/pXO2-) during early, mid, and late log phase were investigated under conditions that simulate those encountered in the host. All of the identified proteins were analyzed by different software algorithms to characterize their predicted mode of secretion and cellular localization. In addition, immunogenic proteins were identified using sera from humans with cutaneous anthrax. A total of 275 extracellular proteins were identified by a combination of LC MS/MS and MALDI-TOF MS. All of the identified proteins were analyzed by SignalP, SecretomeP, PSORT, LipoP, TMHMM, and PROSITE to characterize their predicted mode of secretion, cellular localization, and protein domains. Fifty-three proteins were predicted by SignalP to harbor the cleavable N-terminal signal peptides and were therefore secreted via the classical Sec pathway. Twenty-three proteins were predicted by SecretomeP for secretion by the alternative Sec pathway characterized by the lack of typical export signal. In contrast to SignalP and SecretomeP predictions, PSORT predicted 171 extracellular proteins, 7 cell wall-associated proteins, and 6 cytoplasmic proteins. Moreover, 51 proteins were predicted by LipoP to contain putative Sec signal peptides (38 have SpI sites), lipoprotein signal peptides (13 have SpII sites), and N-terminal membrane helices (9 have transmembrane helices). The TMHMM algorithm predicted 25 membrane-associated proteins with one to ten transmembrane helices. Immunogenic proteins were also identified using sera from patients who have recovered from anthrax. The charge variants (83 and 63 kDa) of protective antigen (PA) were the most immunodominant secreted antigens, followed by charge variants of enolase and transketolase. This is the first description of the time course of protein secretion for the pathogen Bacillus anthracis. Time course studies of protein secretion and accumulation may be relevant in elucidation of the progression of pathogenicity, identification of therapeutics and diagnostic markers, and vaccine development. This study also adds to the continuously growing list of identified Bacillus anthracis secretome proteins.

Bacillus anthracis secretome time course under host-simulated conditions and identification of immunogenic proteins

PubMed Central

Walz, Alexander; Mujer, Cesar V; Connolly, Joseph P; Alefantis, Tim; Chafin, Ryan; Dake, Clarissa; Whittington, Jessica; Kumar, Srikanta P; Khan, Akbar S; DelVecchio, Vito G

2007-01-01

Background The secretion time course of Bacillus anthracis strain RA3R (pXO1+/pXO2-) during early, mid, and late log phase were investigated under conditions that simulate those encountered in the host. All of the identified proteins were analyzed by different software algorithms to characterize their predicted mode of secretion and cellular localization. In addition, immunogenic proteins were identified using sera from humans with cutaneous anthrax. Results A total of 275 extracellular proteins were identified by a combination of LC MS/MS and MALDI-TOF MS. All of the identified proteins were analyzed by SignalP, SecretomeP, PSORT, LipoP, TMHMM, and PROSITE to characterize their predicted mode of secretion, cellular localization, and protein domains. Fifty-three proteins were predicted by SignalP to harbor the cleavable N-terminal signal peptides and were therefore secreted via the classical Sec pathway. Twenty-three proteins were predicted by SecretomeP for secretion by the alternative Sec pathway characterized by the lack of typical export signal. In contrast to SignalP and SecretomeP predictions, PSORT predicted 171 extracellular proteins, 7 cell wall-associated proteins, and 6 cytoplasmic proteins. Moreover, 51 proteins were predicted by LipoP to contain putative Sec signal peptides (38 have SpI sites), lipoprotein signal peptides (13 have SpII sites), and N-terminal membrane helices (9 have transmembrane helices). The TMHMM algorithm predicted 25 membrane-associated proteins with one to ten transmembrane helices. Immunogenic proteins were also identified using sera from patients who have recovered from anthrax. The charge variants (83 and 63 kDa) of protective antigen (PA) were the most immunodominant secreted antigens, followed by charge variants of enolase and transketolase. Conclusion This is the first description of the time course of protein secretion for the pathogen Bacillus anthracis. Time course studies of protein secretion and accumulation may be relevant in elucidation of the progression of pathogenicity, identification of therapeutics and diagnostic markers, and vaccine development. This study also adds to the continuously growing list of identified Bacillus anthracis secretome proteins. PMID:17662140

Understanding the Role of O-GlcNAc Modifications in Plant Development

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

Olszewski, Neil, E.

2011-06-16

This project has contributed towards understanding the role of O-GlcNAc (O-linked N-acetylglucosamine) transferases (OGTs) in plants. Through analyses of single and double mutants, we have investigated the unique and overlapping functions of SECRET AGENT (SEC) and SPINDLY (SPY), the arabidopsis OGTs. This work showed that SEC functions as negative regulators of the long-day flowering pathway. SEC also has a positive role in regulation of rosette. An E. coli co-expression system that allows potential substrates to be co-expressed with and O-GlcNAc modified by SEC was developed. We showed that SEC is a bona fide OGT that modifies itself with single O-linkedmore » GlcNAc(s). Using this system, we tested a number of proteins that were hypothesized to be substrates of SEC and identified a number of substrates include GIGANTEA (GI), a component of the long day flowering pathway. The hypothesis that O-GlcNAc modification controls GI activity was tested by first mapping where E. coli-expressed SEC modifies GI and then assessing the activity of a non-modifiable mutant form of GI. The activity of the mutant form of GI was indistinguishable from that of wild type suggesting that either O-GlcNAc does not regulate GI activity or that additional modification sites exist on GI. In collaboration with Dr. Juan Antonio Garcia at Universidad Autónoma de Madrid the role of O-GlcNAc modification of the plum pox virus coat protein (PPV-CP) was investigated. SEC was shown to O-GlcNAc modify PPV-CP and the modification was shown to facilitate the infection process. E. coli-expressed SEC was shown to modify the same PPV-CP sites that are modified in plants. SEC has a large protein interaction domain called the TPR domain that has been hypothesized to have a role in determining the substrate specificity of the enzyme and/or to regulate its activity. A mutational analysis of the TPR domain did not find evidence for a role in substrate specificity but did obtain evidence that the domain regulates enzyme activity.« less

SNARE-encoding genes VdSec22 and VdSso1 mediate protein secretion required for full virulence in Verticillium dahliae

USDA-ARS?s Scientific Manuscript database

Proteins that mediate cellular and subcellular membrane fusion are key factors in vesicular trafficking in all eukaryotic cells, including the secretion and transport of plant pathogen virulence factors. In this study, we identified vesicle fusion components that included 22 soluble N-ethylmaleimide...

Development of Bacteriocinogenic Strains of Saccharomyces cerevisiae Heterologously Expressing and Secreting the Leaderless Enterocin L50 Peptides L50A and L50B from Enterococcus faecium L50▿

PubMed Central

Basanta, Antonio; Herranz, Carmen; Gutiérrez, Jorge; Criado, Raquel; Hernández, Pablo E.; Cintas, Luis M.

2009-01-01

A segregationally stable expression and secretion vector for Saccharomyces cerevisiae, named pYABD01, was constructed by cloning the yeast gene region encoding the mating pheromone α-factor 1 secretion signal (MFα1s) into the S. cerevisiae high-copy-number expression vector pYES2. The structural genes of the two leaderless peptides of enterocin L50 (EntL50A and EntL50B) from Enterococcus faecium L50 were cloned, separately (entL50A or entL50B) and together (entL50AB), into pYABD01 under the control of the galactose-inducible promoter PGAL1. The generation of recombinant S. cerevisiae strains heterologously expressing and secreting biologically active EntL50A and EntL50B demonstrates the suitability of the MFα1s-containing vector pYABD01 to direct processing and secretion of these antimicrobial peptides through the S. cerevisiae Sec system. PMID:19218405

Bottleneck in secretion of α-amylase in Bacillus subtilis.

PubMed

Yan, Shaomin; Wu, Guang

2017-07-19

Amylase plays an important role in biotechnology industries, and Gram-positive bacterium Bacillus subtilis is a major host to produce heterogeneous α-amylases. However, the secretion stress limits the high yield of α-amylase in B. subtilis although huge efforts have been made to address this secretion bottleneck. In this question-oriented review, every effort is made to answer the following questions, which look simple but are long-standing, through reviewing of literature: (1) Does α-amylase need a specific and dedicated chaperone? (2) What signal sequence does CsaA recognize? (3) Does CsaA require ATP for its operation? (4) Does an unfolded α-amylase is less soluble than a folded one? (5) Does α-amylase aggregate before transporting through Sec secretion system? (6) Is α-amylase sufficient stable to prevent itself from misfolding? (7) Does α-amylase need more disulfide bonds to be stabilized? (8) Which secretion system does PrsA pass through? (9) Is PrsA ATP-dependent? (10) Is PrsA reused after folding of α-amylase? (11) What is the fate of PrsA? (12) Is trigger factor (TF) ATP-dependent? The literature review suggests that not only the most of those questions are still open to answers but also it is necessary to calculate ATP budget in order to better understand how B. subtilis uses its energy for production and secretion.

Selective expression of a sec1/munc18 member in sea urchin eggs and embryos.

PubMed

Leguia, Mariana; Wessel, Gary M

2004-10-01

Regulated secretion is mediated by SNAREs (soluble NSF attachment receptors) and their regulators and effectors, which include the SM (sec1/munc18) family of proteins. Homologs of the SNAREs have been identified in sea urchins, associated with cortical granule exocytosis at fertilization, with membranes of the cleavage furrow, and in secretory cells later in development. To contribute to the understanding of regulated secretion in sea urchins we have cloned the single SM protein homolog from two species of sea urchin, Lytechinus variegatus and Strongylocentrotus purpuratus. In oocytes and eggs, we find that it localizes to the plasma membrane and the cortical region of the egg, consistent with a role in one of the steps leading to cortical granule exocytosis. The protein is also expressed throughout development, enriched in membranes of the cleavage furrow in early embryos, and in cells of the gut in advanced embryos. Furthermore, we find that sec1/munc18 co-localizes with its cognate binding partner syntaxin. Finally, our biochemical analysis shows that the protein associates with rab3 in high molecular weight complexes, suggesting that the exocytotic machinery functions as a multi-protein subunit to mediate regulated secretion in sea urchins. These results will be instrumental in the future to functionally test the SNARE regulators associated with multiple membrane fusion events.

secHsp70 as a tool to approach amyloid-β42 and other extracellular amyloids.

PubMed

De Mena, Lorena; Chhangani, Deepak; Fernandez-Funez, Pedro; Rincon-Limas, Diego E

2017-07-03

Self-association of amyloidogenic proteins is the main pathological trigger in a wide variety of neurodegenerative disorders. These aggregates are deposited inside or outside the cell due to hereditary mutations, environmental exposures or even normal aging. Cumulative evidence indicates that the heat shock chaperone Hsp70 possesses robust neuroprotection against various intracellular amyloids in Drosophila and mouse models. However, its protective role against extracellular amyloids was largely unknown as its presence outside the cells is very limited. Our recent manuscript in PNAS revealed that an engineered form of secreted Hsp70 (secHsp70) is highly protective against toxicity induced by extracellular deposition of the amyloid-β42 (Aβ42) peptide. In this Extra View article, we extend our analysis to other members of the heat shock protein family. We created PhiC31-based transgenic lines for human Hsp27, Hsp40, Hsp60 and Hsp70 and compared their activities in parallel against extracellular Aβ42. Strikingly, only secreted Hsp70 exhibits robust protection against Aβ42-triggered toxicity in the extracellular milieu. These observations indicate that the ability of secHsp70 to suppress Aβ42 insults is quite unique and suggest that targeted secretion of Hsp70 may represent a new therapeutic approach against Aβ42 and other extracellular amyloids. The potential applications of this engineered chaperone are discussed.

Two O-linked N-acetylglucosamine transferase genes of Arabidopsis thaliana L. Heynh. have overlapping functions necessary for gamete and seed development.

PubMed Central

Hartweck, Lynn M; Scott, Cheryl L; Olszewski, Neil E

2002-01-01

The Arabidopsis SECRET AGENT (SEC) and SPINDLY (SPY) proteins are similar to animal O-linked N-acetylglucosamine transferases (OGTs). OGTs catalyze the transfer of N-acetylglucosamine (GlcNAc) from UDP-GlcNAc to Ser/Thr residues of proteins. In animals, O-GlcNAcylation has been shown to affect protein activity, stability, and/or localization. SEC protein expressed in Escherichia coli had autocatalytic OGT activity. To determine the function of SEC in plants, two tDNA insertional mutants were identified and analyzed. Although sec mutant plants did not exhibit obvious phenotypes, sec and spy mutations had a synthetic lethal interaction. This lethality was incompletely penetrant in gametes and completely penetrant postfertilization. The rate of both female and male sec spy gamete transmission was higher in plants heterozygous for both mutations than in plants heterozygous for sec and homozygous for spy. Double-mutant embryos aborted at various stages of development and no double-mutant seedlings were obtained. These results indicate that OGT activity is required during gametogenesis and embryogenesis with lethality occurring when parentally derived SEC, SPY, and/or O-GlcNAcylated proteins become limiting. PMID:12136030

The casein kinases Yck1p and Yck2p act in the secretory pathway, in part, by regulating the Rab exchange factor Sec2p

PubMed Central

Stalder, Danièle; Novick, Peter J.

2016-01-01

Sec2p is a guanine nucleotide exchange factor that activates Sec4p, the final Rab GTPase of the yeast secretory pathway. Sec2p is recruited to secretory vesicles by the upstream Rab Ypt32p acting in concert with phosphatidylinositol-4-phosphate (PI(4)P). Sec2p also binds to the Sec4p effector Sec15p, yet Ypt32p and Sec15p compete against each other for binding to Sec2p. We report here that the redundant casein kinases Yck1p and Yck2p phosphorylate sites within the Ypt32p/Sec15p binding region and in doing so promote binding to Sec15p and inhibit binding to Ypt32p. We show that Yck2p binds to the autoinhibitory domain of Sec2p, adjacent to the PI(4)P binding site, and that addition of PI(4)P inhibits Sec2p phosphorylation by Yck2p. Loss of Yck1p and Yck2p function leads to accumulation of an intracellular pool of the secreted glucanase Bgl2p, as well as to accumulation of Golgi-related structures in the cytoplasm. We propose that Sec2p is phosphorylated after it has been recruited to secretory vesicles and the level of PI(4)P has been reduced. This promotes Sec2p function by stimulating its interaction with Sec15p. Finally, Sec2p is dephosphorylated very late in the exocytic reaction to facilitate recycling. PMID:26700316

MatureP: prediction of secreted proteins with exclusive information from their mature regions.

PubMed

Orfanoudaki, Georgia; Markaki, Maria; Chatzi, Katerina; Tsamardinos, Ioannis; Economou, Anastassios

2017-06-12

More than a third of the cellular proteome is non-cytoplasmic. Most secretory proteins use the Sec system for export and are targeted to membranes using signal peptides and mature domains. To specifically analyze bacterial mature domain features, we developed MatureP, a classifier that predicts secretory sequences through features exclusively computed from their mature domains. MatureP was trained using Just Add Data Bio, an automated machine learning tool. Mature domains are predicted efficiently with ~92% success, as measured by the Area Under the Receiver Operating Characteristic Curve (AUC). Predictions were validated using experimental datasets of mutated secretory proteins. The features selected by MatureP reveal prominent differences in amino acid content between secreted and cytoplasmic proteins. Amino-terminal mature domain sequences have enhanced disorder, more hydroxyl and polar residues and less hydrophobics. Cytoplasmic proteins have prominent amino-terminal hydrophobic stretches and charged regions downstream. Presumably, secretory mature domains comprise a distinct protein class. They balance properties that promote the necessary flexibility required for the maintenance of non-folded states during targeting and secretion with the ability of post-secretion folding. These findings provide novel insight in protein trafficking, sorting and folding mechanisms and may benefit protein secretion biotechnology.

A new potential secretion pathway for recombinant proteins in Bacillus subtilis.

PubMed

Wang, Guangqiang; Xia, Yongjun; Gu, Zhennan; Zhang, Hao; Chen, Yong Q; Chen, Haiqin; Ai, Lianzhong; Chen, Wei

2015-11-10

Secretion of cytoplasmic expressed proteins into growth media has significant advantages. Due to the lack of an outer membrane, Bacillus subtilis is considered as a desirable 'cell factory' for the secretion of recombinant proteins. However, bottlenecks in the classical pathway for the secretion of recombinant proteins limit its use on a wide scale. In this study, we attempted to use four typical non-classically secreted proteins as signals to export three recombinant model proteins to the culture medium. All four non-classically secreted proteins can direct the export of the intrinsically disordered nucleoskeletal-like protein (Nsp). Two of them can guide the secretion of alkaline phosphatase (PhoA). One can lead the secretion of the thermostable β-galactosidase BgaB, which cannot be secreted with the aid of typical Sec-dependent signal peptides. Our results show that the non-classically secreted proteins lead the recombinant proteins to the culture medium, and thus non-classical protein secretion pathways can be exploited as a novel secretion pathway for recombinant proteins.

Diverse C-Terminal Sequences Involved in Flavobacterium johnsoniae Protein Secretion

PubMed Central

Kulkarni, Surashree S.; Zhu, Yongtao; Brendel, Colton J.

2017-01-01

ABSTRACT Flavobacterium johnsoniae and many related bacteria secrete proteins across the outer membrane using the type IX secretion system (T9SS). Proteins secreted by T9SSs have amino-terminal signal peptides for export across the cytoplasmic membrane by the Sec system and carboxy-terminal domains (CTDs) targeting them for secretion across the outer membrane by the T9SS. Most but not all T9SS CTDs belong to the family TIGR04183 (type A CTDs). We functionally characterized diverse CTDs for secretion by the F. johnsoniae T9SS. Attachment of the CTDs from F. johnsoniae RemA, AmyB, and ChiA to the foreign superfolder green fluorescent protein (sfGFP) that had a signal peptide at the amino terminus resulted in secretion across the outer membrane. In each case, approximately 80 to 100 amino acids from the extreme carboxy termini were needed for efficient secretion. Several type A CTDs from distantly related members of the phylum Bacteroidetes functioned in F. johnsoniae, supporting the secretion of sfGFP by the F. johnsoniae T9SS. F. johnsoniae SprB requires the T9SS for secretion but lacks a type A CTD. It has a conserved C-terminal domain belonging to the family TIGR04131, which we refer to as a type B CTD. The CTD of SprB was required for its secretion, but attachment of C-terminal regions of SprB of up to 1,182 amino acids to sfGFP failed to result in secretion. Additional features outside the C-terminal region of SprB may be required for its secretion. IMPORTANCE Type IX protein secretion systems (T9SSs) are common in but limited to members of the phylum Bacteroidetes. Most proteins that are secreted by T9SSs have conserved carboxy-terminal domains that belong to the protein domain family TIGR04183 (type A CTDs) or TIGR04131 (type B CTDs). Here, we identify features of T9SS CTDs of F. johnsoniae that are required for protein secretion and demonstrate that type A CTDs from distantly related members of the phylum function with the F. johnsoniae T9SS to secrete the foreign protein sfGFP. In contrast, type B CTDs failed to target sfGFP for secretion, suggesting a more complex association with the T9SS. PMID:28396348

Fission Yeast Sec3 and Exo70 Are Transported on Actin Cables and Localize the Exocyst Complex to Cell Poles

PubMed Central

Martin, Sophie G.

2012-01-01

The exocyst complex is essential for many exocytic events, by tethering vesicles at the plasma membrane for fusion. In fission yeast, polarized exocytosis for growth relies on the combined action of the exocyst at cell poles and myosin-driven transport along actin cables. We report here the identification of fission yeast Schizosaccharomyces pombe Sec3 protein, which we identified through sequence homology of its PH-like domain. Like other exocyst subunits, sec3 is required for secretion and cell division. Cells deleted for sec3 are only conditionally lethal and can proliferate when osmotically stabilized. Sec3 is redundant with Exo70 for viability and for the localization of other exocyst subunits, suggesting these components act as exocyst tethers at the plasma membrane. Consistently, Sec3 localizes to zones of growth independently of other exocyst subunits but depends on PIP2 and functional Cdc42. FRAP analysis shows that Sec3, like all other exocyst subunits, localizes to cell poles largely independently of the actin cytoskeleton. However, we show that Sec3, Exo70 and Sec5 are transported by the myosin V Myo52 along actin cables. These data suggest that the exocyst holocomplex, including Sec3 and Exo70, is present on exocytic vesicles, which can reach cell poles by either myosin-driven transport or random walk. PMID:22768263

Circulating Neprilysin Clears Brain Amyloid

PubMed Central

Liu, Yinxing; Studzinski, Christa; Beckett, Tina; Murphy, M. Paul; Klein, Ronald L.; Hersh, Louis B.

2010-01-01

The use of the peptidase neprilysin (NEP) as a therapeutic for lowering brain amyloid burden is receiving increasing attention. We have previously demonstrated that peripheral expression of NEP on the surface of hindlimb muscle lowers brain amyloid burden in a transgenic mouse model of Alzheimer’s disease. In this study we now show that using adeno-associated virus expressing a soluble secreted form of NEP (secNEP-AAV8), NEP secreted into plasma is effective in clearing brain Aβ. Soluble NEP expression in plasma was sustained over the 3-month time period it was measured. Secreted NEP decreased plasma Aβ by 30%, soluble brain Aβ by ~28%, insoluble brain Aβ by ~55%, and Aβ oligomers by 12%. This secNEP did not change plasma levels of substance P or bradykinin, nor did it alter blood pressure. No NEP was detected in CSF, nor did the AAV virus produce brain expression of NEP. Thus the lowering of brain Aβ was due to plasma NEP which altered blood-brain Aβ transport dynamics. Expressing NEP in plasma provides a convenient way to monitor enzyme activity during the course of its therapeutic testing. PMID:20558294

Use of the Yeast Pichia pastoris as an Expression Host for Secretion of Enterocin L50, a Leaderless Two-Peptide (L50A and L50B) Bacteriocin from Enterococcus faecium L50▿

PubMed Central

Basanta, Antonio; Gómez-Sala, Beatriz; Sánchez, Jorge; Diep, Dzung B.; Herranz, Carmen; Hernández, Pablo E.; Cintas, Luis M.

2010-01-01

In this work, we report the expression and secretion of the leaderless two-peptide (EntL50A and EntL50B) bacteriocin enterocin L50 from Enterococcus faecium L50 by the methylotrophic yeast Pichia pastoris X-33. The bacteriocin structural genes entL50A and entL50B were fused to the Saccharomyces cerevisiae gene region encoding the mating pheromone α-factor 1 secretion signal (MFα1s) and cloned, separately and together (entL50AB), into the P. pastoris expression and secretion vector pPICZαA, which contains the methanol-inducible alcohol oxidase promoter (PAOX1) to express the fusion genes. After transfer into the yeast, the recombinant plasmids were integrated into the genome, resulting in three bacteriocinogenic yeast strains able to produce and secrete the individual bacteriocin peptides EntL50A and EntL50B separately and together. The secretion was efficiently directed by MFα1s through the Sec system, and the precursor peptides were found to be correctly processed to form mature and active bacteriocin peptides. The present work describes for the first time the heterologous expression and secretion of a two-peptide non-pediocin-like bacteriocin by a yeast. PMID:20348300

High-Resolution Genetics Identifies the Lipid Transfer Protein Sec14p as Target for Antifungal Ergolines

PubMed Central

Cotesta, Simona; Perruccio, Francesca; Knapp, Britta; Fu, Yue; Studer, Christian; Pries, Verena; Riedl, Ralph; Helliwell, Stephen B.; Petrovic, Katarina T.; Movva, N. Rao; Sanglard, Dominique; Tao, Jianshi; Hoepfner, Dominic

2016-01-01

Invasive infections by fungal pathogens cause more deaths than malaria worldwide. We found the ergoline compound NGx04 in an antifungal screen, with selectivity over mammalian cells. High-resolution chemogenomics identified the lipid transfer protein Sec14p as the target of NGx04 and compound-resistant mutations in Sec14p define compound-target interactions in the substrate binding pocket of the protein. Beyond its essential lipid transfer function in a variety of pathogenic fungi, Sec14p is also involved in secretion of virulence determinants essential for the pathogenicity of fungi such as Cryptococcus neoformans, making Sec14p an attractive antifungal target. Consistent with this dual function, we demonstrate that NGx04 inhibits the growth of two clinical isolates of C. neoformans and that NGx04-related compounds have equal and even higher potency against C. neoformans. Furthermore NGx04 analogues showed fungicidal activity against a fluconazole resistant C. neoformans strain. In summary, we present genetic evidence that NGx04 inhibits fungal Sec14p and initial data supporting NGx04 as a novel antifungal starting point. PMID:27855158

Extracellular proteome analysis of Leptospira interrogans serovar Lai.

PubMed

Zeng, Lingbing; Zhang, Yunyi; Zhu, Yongzhang; Yin, Haidi; Zhuang, Xuran; Zhu, Weinan; Guo, Xiaokui; Qin, Jinhong

2013-10-01

Abstract Leptospirosis is one of the most important zoonoses. Leptospira interrogans serovar Lai is a pathogenic spirochete that is responsible for leptospirosis. Extracellular proteins play an important role in the pathogenicity of this bacterium. In this study, L. interrogans serovar Lai was grown in protein-free medium; the supernatant was collected and subsequently analyzed as the extracellular proteome. A total of 66 proteins with more than two unique peptides were detected by MS/MS, and 33 of these were predicted to be extracellular proteins by a combination of bioinformatics analyses, including Psortb, cello, SoSuiGramN and SignalP. Comparisons of the transcriptional levels of these 33 genes between in vivo and in vitro conditions revealed that 15 genes were upregulated and two genes were downregulated in vivo compared to in vitro. A BLAST search for the components of secretion system at the genomic and proteomic levels revealed the presence of the complete type I secretion system and type II secretion system in this strain. Moreover, this strain also exhibits complete Sec translocase and Tat translocase systems. The extracellular proteome analysis of L. interrogans will supplement the previously generated whole proteome data and provide more information for studying the functions of specific proteins in the infection process and for selecting candidate molecules for vaccines or diagnostic tools for leptospirosis.

The Structure of RalF, an ADP-Ribosylation Factor Guanine Nucleotide Exchange Factor from Legionella pneumophila, Reveals the Presence of a Cap over the Active Site

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

Amor,J.; Swails, J.; Zhu, X.

2005-01-01

The Legionella pneumophila protein RalF is secreted into host cytosol via the Dot/Icm type IV transporter where it acts to recruit ADP-ribosylation factor (Arf) to pathogen-containing phagosomes in the establishment of a replicative organelle. The presence in RalF of the Sec7 domain, present in all Arf guanine nucleotide exchange factors, has suggested that recruitment of Arf is an early step in pathogenesis. We have determined the crystal structure of RalF and of the isolated Sec7 domain and found that RalF is made up of two domains. The Sec7 domain is homologous to mammalian Sec7 domains. The C-terminal domain forms amore » cap over the active site in the Sec7 domain and contains a conserved folding motif, previously observed in adaptor subunits of vesicle coat complexes. The importance of the capping domain and of the glutamate in the 'glutamic finger,' conserved in all Sec7 domains, to RalF functions was examined using three different assays. These data highlight the functional importance of domains other than Sec7 in Arf guanine nucleotide exchange factors to biological activities and suggest novel mechanisms of regulation of those activities.« less

Circulating neprilysin clears brain amyloid.

PubMed

Liu, Yinxing; Studzinski, Christa; Beckett, Tina; Murphy, M Paul; Klein, Ronald L; Hersh, Louis B

2010-10-01

The use of the peptidase neprilysin (NEP) as a therapeutic for lowering brain amyloid burden is receiving increasing attention. We have previously demonstrated that peripheral expression of NEP on the surface of hindlimb muscle lowers brain amyloid burden in a transgenic mouse model of Alzheimer's disease. In this study we now show that using adeno-associated virus expressing a soluble secreted form of NEP (secNEP-AAV8), NEP secreted into plasma is effective in clearing brain Abeta. Soluble NEP expression in plasma was sustained over the 3-month time period it was measured. Secreted NEP decreased plasma Abeta by 30%, soluble brain Abeta by approximately 28%, insoluble brain Abeta by approximately 55%, and Abeta oligomersby 12%. This secNEP did not change plasma levels of substance P or bradykinin, nor did it alter blood pressure. No NEP was detected in CSF, nor did the AAV virus produce brain expression of NEP. Thus the lowering of brain Abeta was due to plasma NEP which altered blood-brain Abeta transport dynamics. Expressing NEP in plasma provides a convenient way to monitor enzyme activity during the course of its therapeutic testing. Copyright 2010 Elsevier Inc. All rights reserved.

Comparative proteome analysis of laboratory grown Brucella abortus 2308 and Brucella melitensis 16M.

PubMed

Eschenbrenner, Michel; Horn, Troy A; Wagner, Mary Ann; Mujer, Cesar V; Miller-Scandle, Tabbi L; DelVecchio, Vito G

2006-07-01

Brucella species are pathogenic agents that cause brucellosis, a debilitating zoonotic disease that affects a large variety of domesticated animals and humans. Brucella melitensis and Brucella abortus are considered major health threats because of their highly infectious nature and worldwide occurrence. The availability of the annotated genomes for these two species has allowed a comparative proteomics study of laboratory grown B. melitensis 16M and B. abortus 2308 by two-dimensional (2-D) gel electrophoresis and peptide mass fingerprinting. Computer-assisted analysis of the different 2-D gel images of strains 16M and 2308 revealed significant quantitative and qualitative differences in their protein expression patterns. Proteins involved in membrane transport, particularly the high affinity amino acids binding proteins, and those involved in Sec-dependent secretion systems related to type IV and type V secretion systems, were differentially expressed. Differential expression of these proteins may be responsible for conferring specific host preference in the two strains 2308 and 16M.

Mechanisms of ribosome stalling by SecM at multiple elongation steps

PubMed Central

Zhang, Jun; Pan, Xijiang; Yan, Kaige; Sun, Shan; Gao, Ning; Sui, Sen-Fang

2015-01-01

Regulation of translating ribosomes is a major component of gene expression control network. In Escherichia coli, ribosome stalling by the C-terminal arrest sequence of SecM regulates the SecA-dependent secretion pathway. Previous studies reported many residues of SecM peptide and ribosome exit tunnel are critical for stalling. However, the underlying molecular mechanism is still not clear at the atomic level. Here, we present two cryo-EM structures of the SecM-stalled ribosomes at 3.3–3.7 Å resolution, which reveal two different stalling mechanisms at distinct elongation steps of the translation cycle: one is due to the inactivation of ribosomal peptidyl-transferase center which inhibits peptide bond formation with the incoming prolyl-tRNA; the other is the prolonged residence of the peptidyl-RNA at the hybrid A/P site which inhibits the full-scale tRNA translocation. These results demonstrate an elegant control of translation cycle by regulatory peptides through a continuous, dynamic reshaping of the functional center of the ribosome. DOI: http://dx.doi.org/10.7554/eLife.09684.001 PMID:26670735

Stimulus-secretion coupling in chromaffin cells isolated from bovine adrenal medulla

PubMed Central

Schneider, Allan S.; Herz, Ruth; Rosenheck, Kurt

1977-01-01

Bovine adrenal chromaffin cells were isolated by removal of the cortex and sequential collagenase digestion of the medulla. The catecholamine secretory function of these cells was characterized with respect to acetylcholine stimulation, cation requirements, and cytoskeletal elements. The dose-response curve for stimulated release had its half-maximum value at 10-5 M acetylcholine, and maximum secretion was on the average 7 times that of control basal secretion. The differential release of epinephrine versus norepinephrine after stimulation with 0.1 mM acetylcholine occurred in proportion to their distribution in the cell suspension. The cholinergic receptors were found to be predominantly nicotinic. The kinetics of catecholamine release were rapid, with significant secretion occurring in less than 60 sec and 85% of maximum secretion within 5 min. A critical requirement for calcium in the extracellular medium was demonstrated, and 80% of maximum secretion was achieved at physiologic calcium concentrations. Stimulation by excess potassium (65 mM KCl) also induced catecholamine secretion which differed from acetylcholine stimulation in being less potent, in having a different dependence on calcium concentration, and in its response to the local anesthetic tetracaine. Tetracaine, which is thought to inhibit membrane cation permeability, was able to block acetylcholine-stimulated but not KCl-stimulated secretion. The microtubule disrupting agent vinblastine was able to block catecholamine release whereas the microfilament disrupter cytochalasin B had little effect. The results show the isolated bovine chromaffin cells to be viable, functioning, and available in large quantity. These cells now provide an excellent system for studying cell surface regulation of hormone and neurotransmitter release. PMID:270738

Structural Analysis of N- and O-glycans Using ZIC-HILIC/Dialysis Coupled to NMR Detection

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

Qu, Yi; Feng, Ju; Deng, Shuang

2014-11-19

Protein glycosylation, an important and complex post-translational modification (PTM), is involved in various biological processes including the receptor-ligand and cell-cell interaction, and plays a crucial role in many biological functions. However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation,) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. In this study, we employed SEC, Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC), and ZIC-HILIC coupled with dialysis strategies to enrich the glycopeptides from the pronase E digests of RNase B, followed by NMR analysis ofmore » the glycoconjugate. Our results suggest that the ZIC-HILIC enrichment coupled with dialysis is the most efficient, which was thus applied to the analysis of biological complex sample, the pronase E digest of the secreted proteins from the fungi Aspergillus niger. The NMR spectra revealed that the secreted proteins from A. niger contain both N-linked glycans with a high-mannose core and O-linked glycans bearing mannose and glucose with 1->3 and 1->6 linkages. In all, our study provides compelling evidence that ZIC-HILIC separation coupled to dialysis is superior to the commonly used SEC separation to prepare glycopeptides for the downstream NMR analysis, which could greatly facilitate the future NMR-based glycoproteomics research.« less

Cystic fibrosis airway secretions exhibit mucin hyperconcentration and increased osmotic pressure

PubMed Central

Henderson, Ashley G.; Ehre, Camille; Button, Brian; Abdullah, Lubna H.; Cai, Li-Heng; Leigh, Margaret W.; DeMaria, Genevieve C.; Matsui, Hiro; Donaldson, Scott H.; Davis, C. William; Sheehan, John K.; Boucher, Richard C.; Kesimer, Mehmet

2014-01-01

The pathogenesis of mucoinfective lung disease in cystic fibrosis (CF) patients likely involves poor mucus clearance. A recent model of mucus clearance predicts that mucus flow depends on the relative mucin concentration of the mucus layer compared with that of the periciliary layer; however, mucin concentrations have been difficult to measure in CF secretions. Here, we have shown that the concentration of mucin in CF sputum is low when measured by immunologically based techniques, and mass spectrometric analyses of CF mucins revealed mucin cleavage at antibody recognition sites. Using physical size exclusion chromatography/differential refractometry (SEC/dRI) techniques, we determined that mucin concentrations in CF secretions were higher than those in normal secretions. Measurements of partial osmotic pressures revealed that the partial osmotic pressure of CF sputum and the retained mucus in excised CF lungs were substantially greater than the partial osmotic pressure of normal secretions. Our data reveal that mucin concentration cannot be accurately measured immunologically in proteolytically active CF secretions; mucins are hyperconcentrated in CF secretions; and CF secretion osmotic pressures predict mucus layer–dependent osmotic compression of the periciliary liquid layer in CF lungs. Consequently, mucin hypersecretion likely produces mucus stasis, which contributes to key infectious and inflammatory components of CF lung disease. PMID:24892808

Homogeneity and heterogeneity in amylase production by Bacillus subtilis under different growth conditions.

PubMed

Ploss, Tina N; Reilman, Ewoud; Monteferrante, Carmine G; Denham, Emma L; Piersma, Sjouke; Lingner, Anja; Vehmaanperä, Jari; Lorenz, Patrick; van Dijl, Jan Maarten

2016-03-29

Bacillus subtilis is an important cell factory for the biotechnological industry due to its ability to secrete commercially relevant proteins in large amounts directly into the growth medium. However, hyper-secretion of proteins, such as α-amylases, leads to induction of the secretion stress-responsive CssR-CssS regulatory system, resulting in up-regulation of the HtrA and HtrB proteases. These proteases degrade misfolded proteins secreted via the Sec pathway, resulting in a loss of product. The aim of this study was to investigate the secretion stress response in B. subtilis 168 cells overproducing the industrially relevant α-amylase AmyM from Geobacillus stearothermophilus, which was expressed from the strong promoter P(amyQ)-M. Here we show that activity of the htrB promoter as induced by overproduction of AmyM was "noisy", which is indicative for heterogeneous activation of the secretion stress pathway. Plasmids were constructed to allow real-time analysis of P(amyQ)-M promoter activity and AmyM production by, respectively, transcriptional and out-of-frame translationally coupled fusions with gfpmut3. Our results show the emergence of distinct sub-populations of high- and low-level AmyM-producing cells, reflecting heterogeneity in the activity of P(amyQ)-M. This most likely explains the heterogeneous secretion stress response. Importantly, more homogenous cell populations with regard to P(amyQ)-M activity were observed for the B. subtilis mutant strain 168degUhy32, and the wild-type strain 168 under optimized growth conditions. Expression heterogeneity of secretory proteins in B. subtilis can be suppressed by degU mutation and optimized growth conditions. Further, the out-of-frame translational fusion of a gene for a secreted target protein and gfp represents a versatile tool for real-time monitoring of protein production and opens novel avenues for Bacillus production strain improvement.

Protein secretion through autotransporter and two-partner pathways.

PubMed

Jacob-Dubuisson, Françoise; Fernandez, Rachel; Coutte, Loic

2004-11-11

Two distinct protein secretion pathways, the autotransporter (AT) and the two-partner secretion (TPS) pathways are characterized by their apparent simplicity. Both are devoted to the translocation across the outer membrane of mostly large proteins or protein domains. As implied by their name, AT proteins contain their own transporter domain, covalently attached to the C-terminal extremity of the secreted passenger domain, while TPS systems are composed of two separate proteins, with TpsA being the secreted protein and TpsB its specific transporter. In both pathways, the secreted proteins are exported in a Sec-dependent manner across the inner membrane, after which they cross the outer membrane with the help of their cognate transporters. The AT translocator domains and the TpsB proteins constitute distinct families of protein-translocating, outer membrane porins of Gram-negative bacteria. Both types of transporters insert into the outer membrane as beta-barrel proteins possibly forming oligomeric pores in the case of AT and serve as conduits for their cognate secreted proteins or domains across the outer membrane. Translocation appears to be folding-sensitive in both pathways, indicating that AT passenger domains and TpsA proteins cross the periplasm and the outer membrane in non-native conformations and fold progressively at the cell surface. A major difference between AT and TPS pathways arises from the manner by which specificity is established between the secreted protein and its transporter. In AT, the covalent link between the passenger and the translocator domains ensures the translocation of the former without the need for a specific molecular recognition between the two modules. In contrast, the TPS pathway has solved the question of specific recognition between the TpsA proteins and their transporters by the addition to the TpsA proteins of an N-proximal module, the conserved TPS domain, which represents a hallmark of the TPS pathway.

Reconstruction of the bifidobacterial pan-secretome reveals the network of extracellular interactions between bifidobacteria and the infant gut.

PubMed

Lugli, Gabriele Andrea; Mancino, Walter; Milani, Christian; Duranti, Sabrina; Turroni, Francesca; van Sinderen, Douwe; Ventura, Marco

2018-06-08

The repertoire of secreted proteins decoded by a microorganism represents proteins released from or associated with the cell's surface. In gut commensals, such as bifidobacteria, these proteins are perceived to be functionally relevant as they regulate the interaction with the gut environment. In the current study, we have screened the predicted proteome of over 300 bifidobacterial strains amongst the currently recognized bifidobacterial species to generate a comprehensive database encompassing bifidobacterial extracellular proteins. A glycobiome analysis of this predicted bifidobacterial secretome revealed that a correlation exists between particular bifidobacterial species and their capability to hydrolyze HMOs and intestinal glyconjugates such as mucin. Furthermore, exploration of metatranscriptomic datasets of the infant gut microbiota allowed the evaluation of the expression of bifidobacterial genes encoding extracellular proteins, represented by ABC transporter substrate-binding proteins and glycoside hydrolases enzymes involved in the degradation of human milk oligosaccharides and mucin. Overall, this study provides insights into how bifidobacteria interact with their natural yet highly complex environment, the infant gut. Importance The ecological success of bifidobacteria relies on the activity of extracellular proteins that are involved in the metabolism of nutrients and the interaction with the environment. To date, information on secreted proteins encoded by bifidobacteria are incomplete and just related to few species. In this study, we reconstructed the bifidobacterial pan-secretome, revealing extracellular proteins that modulate the interaction of bifidobacteria with their natural environment. Furthermore, a survey of secretion system between bifidobacterial genomes allowed the identification of a conserved Sec-dependent secretion machinery in all the analyzed genomes and the Tat protein translocation system in the chromosomes of 23 strains belonging to Bifidobacterium longum subsp. longum and Bifidobacterium aesculapii . Copyright © 2018 American Society for Microbiology.

JVirGel 2.0: computational prediction of proteomes separated via two-dimensional gel electrophoresis under consideration of membrane and secreted proteins.

PubMed

Hiller, Karsten; Grote, Andreas; Maneck, Matthias; Münch, Richard; Jahn, Dieter

2006-10-01

After the publication of JVirGel 1.0 in 2003 we got many requests and suggestions from the proteomics community to further improve the performance of the software and to add additional useful new features. The integration of the PrediSi algorithm for the prediction of signal peptides for the Sec-dependent protein export into JVirGel 2.0 allows the exclusion of most exported preproteins from calculated proteomic maps and provides the basis for the calculation of Sec-based secretomes. A tool for the identification of transmembrane helices carrying proteins (JCaMelix) and the prediction of the corresponding membrane proteome was added. Finally, in order to directly compare experimental and calculated proteome data, a function to overlay and evaluate predicted and experimental two-dimensional gels was included. JVirGel 2.0 is freely available as precompiled package for the installation on Windows or Linux operating systems. Furthermore, there is a completely platform-independent Java version available for download. Additionally, we provide a Java Server Pages based version of JVirGel 2.0 which can be operated in nearly all web browsers. All versions are accessible at http://www.jvirgel.de

Selective export of autotaxin from the endoplasmic reticulum.

PubMed

Lyu, Lin; Wang, Baolu; Xiong, Chaoyang; Zhang, Xiaotian; Zhang, Xiaoyan; Zhang, Junjie

2017-04-28

Autotaxin (ATX) or ectonucleotide pyrophosphatase/phosphodiesterase 2 (ENPP2) is a secretory glycoprotein and functions as the key enzyme for lysophosphatidic acid generation. The mechanism of ATX protein trafficking is largely unknown. Here, we demonstrated that p23, a member of the p24 protein family, was the protein-sorting receptor required for endoplasmic reticulum (ER) export of ATX. A di-phenylalanine (Phe-838/Phe-839) motif in the human ATX C-terminal region was identified as a transport signal essential for the ATX-p23 interaction. Knockdown of individual Sec24 isoforms by siRNA revealed that ER export of ATX was impaired only if Sec24C was down-regulated. These results suggest that ATX is selectively exported from the ER through a p23, Sec24C-dependent pathway. In addition, it was found that AKT signaling played a role in ATX secretion regulation to facilitate ATX ER export by enhancing the nuclear factor of activated T cell-mediated p23 expression. Furthermore, the di-hydrophobic amino acid motifs (FY) also existed in the C-terminal regions of human ENPP1 and ENPP3. Such a p23, Sec24C-dependent selective ER export mechanism is conserved among these ENPP family members. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Immunocytochemical localization of chymase to cytoplasmic vesicles after rat peritoneal mast cell stimulation by compound 48/80.

PubMed

Login, G R; Aoki, M; Yamakawa, M; Lunardi, L O; Digenis, E C; Tanda, N; Schwartz, L B; Dvorak, A M

1997-10-01

The subcellular events responsible for release of mediators by mast cells may help to clarify roles for mast cells in health and disease. In this study we show that the granule-associated protease chymase is also within cytoplasmic vesicles in appropriately stimulated rat peritoneal mast cells. Rat peritoneal mast cells were recovered before or 1-10 sec after exposure to the secretogogue compound 48/80 (10 micrograms/ml) and then were examined by radioimmunoassay to quantify histamine release or were processed, using routine methods for postembedding immunoelectron microscopy, to identify the subcellular localization of chymase. In comparison to unstimulated cells, compound 48/80 stimulated cells in two independent experiments showed an increase (15%, 28%) in the surface area of the cell and a decrease (12%, 6%) in the surface area of the total granule compartment before degranulation channel formation. These global cellular changes occurred in a background of transient but significant (p < 0.01) increases in the area and number of chymase-immunoreactive vesicles per microns2 cytoplasm. These changes were detectable at 5 or 7 sec after stimulation with compound 48/80 but returned to near prestimulation levels by 9 or 10 sec after addition of compound 48/80 (total cumulative histamine release was 28% by 8 sec and 47% by 14 sec). These observations suggest that vesicles participate in the early stages of regulated secretion of chymase from rat peritoneal mast cells.

Nanosized UCMSC-derived extracellular vesicles but not conditioned medium exclusively inhibit the inflammatory response of stimulated T cells: implications for nanomedicine.

PubMed

Monguió-Tortajada, Marta; Roura, Santiago; Gálvez-Montón, Carolina; Pujal, Josep Maria; Aran, Gemma; Sanjurjo, Lucía; Franquesa, Marcel la; Sarrias, Maria-Rosa; Bayes-Genis, Antoni; Borràs, Francesc E

2017-01-01

Undesired immune responses have drastically hampered outcomes after allogeneic organ transplantation and cell therapy, and also lead to inflammatory diseases and autoimmunity. Umbilical cord mesenchymal stem cells (UCMSCs) have powerful regenerative and immunomodulatory potential, and their secreted extracellular vesicles (EVs) are envisaged as a promising natural source of nanoparticles to increase outcomes in organ transplantation and control inflammatory diseases. However, poor EV preparations containing highly-abundant soluble proteins may mask genuine vesicular-associated functions and provide misleading data. Here, we used Size-Exclusion Chromatography (SEC) to successfully isolate EVs from UCMSCs-conditioned medium. These vesicles were defined as positive for CD9, CD63, CD73 and CD90, and their size and morphology characterized by NTA and cryo-EM. Their immunomodulatory potential was determined in polyclonal T cell proliferation assays, analysis of cytokine profiles and in the skewing of monocyte polarization. In sharp contrast to the non-EV containing fractions, to the complete conditioned medium and to ultracentrifuged pellet, SEC-purified EVs from UCMSCs inhibited T cell proliferation, resembling the effect of parental UCMSCs. Moreover, while SEC-EVs did not induce cytokine response, the non-EV fractions, conditioned medium and ultracentrifuged pellet promoted the secretion of pro-inflammatory cytokines by polyclonally stimulated T cells and supported Th17 polarization. In contrast, EVs did not induce monocyte polarization, but the non-EV fraction induced CD163 and CD206 expression and TNF-α production in monocytes. These findings increase the growing evidence confirming that EVs are an active component of MSC's paracrine immunosuppressive function and affirm their potential for therapeutics in nanomedicine. In addition, our results highlight the importance of well-purified and defined preparations of MSC-derived EVs to achieve the immunosuppressive effect.

RNA Silencing of Exocyst Genes in the Stigma Impairs the Acceptance of Compatible Pollen in Arabidopsis1[OPEN

PubMed Central

Safavian, Darya; Indriolo, Emily; Chapman, Laura; Ahmed, Abdalla

2015-01-01

Initial pollen-pistil interactions in the Brassicaceae are regulated by rapid communication between pollen grains and stigmatic papillae and are fundamentally important, as they are the first step toward successful fertilization. The goal of this study was to examine the requirement of exocyst subunits, which function in docking secretory vesicles to sites of polarized secretion, in the context of pollen-pistil interactions. One of the exocyst subunit genes, EXO70A1, was previously identified as an essential factor in the stigma for the acceptance of compatible pollen in Arabidopsis (Arabidopsis thaliana) and Brassica napus. We hypothesized that EXO70A1, along with other exocyst subunits, functions in the Brassicaceae dry stigma to deliver cargo-bearing secretory vesicles to the stigmatic papillar plasma membrane, under the pollen attachment site, for pollen hydration and pollen tube entry. Here, we investigated the functions of exocyst complex genes encoding the remaining seven subunits, SECRETORY3 (SEC3), SEC5, SEC6, SEC8, SEC10, SEC15, and EXO84, in Arabidopsis stigmas following compatible pollinations. Stigma-specific RNA-silencing constructs were used to suppress the expression of each exocyst subunit individually. The early postpollination stages of pollen grain adhesion, pollen hydration, pollen tube penetration, seed set, and overall fertility were analyzed in the transgenic lines to evaluate the requirement of each exocyst subunit. Our findings provide comprehensive evidence that all eight exocyst subunits are necessary in the stigma for the acceptance of compatible pollen. Thus, this work implicates a fully functional exocyst complex as a component of the compatible pollen response pathway to promote pollen acceptance. PMID:26443677

O-GlcNAcylation of master growth repressor DELLA by SECRET AGENT modulates multiple signaling pathways in Arabidopsis

PubMed Central

Zentella, Rodolfo; Hu, Jianhong; Hsieh, Wen-Ping; Matsumoto, Peter A.; Dawdy, Andrew; Barnhill, Benjamin; Oldenhof, Harriëtte; Hartweck, Lynn M.; Maitra, Sushmit; Thomas, Stephen G.; Cockrell, Shelley; Boyce, Michael; Shabanowitz, Jeffrey; Hunt, Donald F.; Olszewski, Neil E.; Sun, Tai-ping

2016-01-01

The DELLA family of transcription regulators functions as master growth repressors in plants by inhibiting phytohormone gibberellin (GA) signaling in response to developmental and environmental cues. DELLAs also play a central role in mediating cross-talk between GA and other signaling pathways via antagonistic direct interactions with key transcription factors. However, how these crucial protein–protein interactions can be dynamically regulated during plant development remains unclear. Here, we show that DELLAs are modified by the O-linked N-acetylglucosamine (O-GlcNAc) transferase (OGT) SECRET AGENT (SEC) in Arabidopsis. O-GlcNAcylation of the DELLA protein REPRESSOR OF ga1-3 (RGA) inhibits RGA binding to four of its interactors—PHYTOCHROME-INTERACTING FACTOR3 (PIF3), PIF4, JASMONATE-ZIM DOMAIN1, and BRASSINAZOLE-RESISTANT1 (BZR1)—that are key regulators in light, jasmonate, and brassinosteroid signaling pathways, respectively. Consistent with this, the sec-null mutant displayed reduced responses to GA and brassinosteroid and showed decreased expression of several common target genes of DELLAs, BZR1, and PIFs. Our results reveal a direct role of OGT in repressing DELLA activity and indicate that O-GlcNAcylation of DELLAs provides a fine-tuning mechanism in coordinating multiple signaling activities during plant development. PMID:26773002

O-GlcNAcylation of master growth repressor DELLA by SECRET AGENT modulates multiple signaling pathways in Arabidopsis.

PubMed

Zentella, Rodolfo; Hu, Jianhong; Hsieh, Wen-Ping; Matsumoto, Peter A; Dawdy, Andrew; Barnhill, Benjamin; Oldenhof, Harriëtte; Hartweck, Lynn M; Maitra, Sushmit; Thomas, Stephen G; Cockrell, Shelley; Boyce, Michael; Shabanowitz, Jeffrey; Hunt, Donald F; Olszewski, Neil E; Sun, Tai-Ping

2016-01-15

The DELLA family of transcription regulators functions as master growth repressors in plants by inhibiting phytohormone gibberellin (GA) signaling in response to developmental and environmental cues. DELLAs also play a central role in mediating cross-talk between GA and other signaling pathways via antagonistic direct interactions with key transcription factors. However, how these crucial protein-protein interactions can be dynamically regulated during plant development remains unclear. Here, we show that DELLAs are modified by the O-linked N-acetylglucosamine (O-GlcNAc) transferase (OGT) SECRET AGENT (SEC) in Arabidopsis. O-GlcNAcylation of the DELLA protein REPRESSOR OF ga1-3 (RGA) inhibits RGA binding to four of its interactors-PHYTOCHROME-INTERACTING FACTOR3 (PIF3), PIF4, JASMONATE-ZIM DOMAIN1, and BRASSINAZOLE-RESISTANT1 (BZR1)-that are key regulators in light, jasmonate, and brassinosteroid signaling pathways, respectively. Consistent with this, the sec-null mutant displayed reduced responses to GA and brassinosteroid and showed decreased expression of several common target genes of DELLAs, BZR1, and PIFs. Our results reveal a direct role of OGT in repressing DELLA activity and indicate that O-GlcNAcylation of DELLAs provides a fine-tuning mechanism in coordinating multiple signaling activities during plant development. © 2016 Zentella et al.; Published by Cold Spring Harbor Laboratory Press.

Type IX secretion: the generation of bacterial cell surface coatings involved in virulence, gliding motility and the degradation of complex biopolymers.

PubMed

Veith, Paul D; Glew, Michelle D; Gorasia, Dhana G; Reynolds, Eric C

2017-10-01

The Type IX secretion system (T9SS) is present in over 1000 sequenced species/strains of the Fibrobacteres-Chlorobi-Bacteroidetes superphylum. Proteins secreted by the T9SS have an N-terminal signal peptide for translocation across the inner membrane via the SEC translocon and a C-terminal signal for secretion across the outer membrane via the T9SS. Nineteen protein components of the T9SS have been identified including three, SigP, PorX and PorY that are involved in regulation. The inner membrane proteins PorL and PorM and the outer membrane proteins PorK and PorN interact and a complex comprising PorK and PorN forms a large ring structure of 50 nm in diameter. PorU, PorV, PorQ and PorZ form an attachment complex on the cell surface of the oral pathogen, Porphyromonas gingivalis. P. gingivalis T9SS substrates bind to PorV suggesting that after translocation PorV functions as a shuttle protein to deliver T9SS substrates to the attachment complex. The PorU component of the attachment complex is a novel Gram negative sortase which catalyses the cleavage of the C-terminal signal and conjugation of the protein substrates to lipopolysaccharide, anchoring them to the cell surface. This review presents an overview of the T9SS focusing on the function of T9SS substrates and machinery components. © 2017 John Wiley & Sons Ltd.

77 FR 16569 - Privacy Act of 1974: Systems of Records

Federal Register 2010, 2011, 2012, 2013, 2014

2012-03-21

... ``Fitness Center Membership, Payment, and Fitness Records (SEC-48)'', last published in the Federal Register... existing systems of records, ``Administrative Audit System (SEC-14)'' and ``Fitness Center Membership, Payment, and Fitness Records (SEC- 48)''. As described in the last published notice, the Administrative...

Dietary flavonoid fisetin increases abundance of high-molecular-mass hyaluronan conferring resistance to prostate oncogenesis.

PubMed

Lall, Rahul K; Syed, Deeba N; Khan, Mohammad Imran; Adhami, Vaqar M; Gong, Yuansheng; Lucey, John A; Mukhtar, Hasan

2016-09-01

We and others have shown previously that fisetin, a plant flavonoid, has therapeutic potential against many cancer types. Here, we examined the probable mechanism of its action in prostate cancer (PCa) using a global metabolomics approach. HPLC-ESI-MS analysis of tumor xenografts from fisetin-treated animals identified several metabolic targets with hyaluronan (HA) as the most affected. Efficacy of fisetin on HA was then evaluated in vitro and also in vivo in the transgenic TRAMP mouse model of PCa. Size exclusion chromatography-multiangle laser light scattering (SEC-MALS) was performed to analyze the molar mass (Mw) distribution of HA. Fisetin treatment downregulated intracellular and secreted HA levels both in vitro and in vivo Fisetin inhibited HA synthesis and degradation enzymes, which led to cessation of HA synthesis and also repressed the degradation of the available high-molecular-mass (HMM)-HA. SEC-MALS analysis of intact HA fragment size revealed that cells and animals have more abundance of HMM-HA and less of low-molecular-mass (LMM)-HA upon fisetin treatment. Elevated HA levels have been shown to be associated with disease progression in certain cancer types. Biological responses triggered by HA mainly depend on the HA polymer length where HMM-HA represses mitogenic signaling and has anti-inflammatory properties whereas LMM-HA promotes proliferation and inflammation. Similarly, Mw analysis of secreted HA fragment size revealed less HMM-HA is secreted that allowed more HMM-HA to be retained within the cells and tissues. Our findings establish that fisetin is an effective, non-toxic, potent HA synthesis inhibitor, which increases abundance of antiangiogenic HMM-HA and could be used for the management of PCa. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

A computerized system to measure and predict air quality for emission control

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

Crooks, G.; Ciccone, A.; Frattolillo, P.

1997-12-31

A Supplementary Emission Control (SEC) system has been developed on behalf of the Association Industrielle de l`Est de Montreal (AIEM). The objective of the SEC is to avoid exceedences of the Montreal Urban Community (MUC) 24 hour ambient Air Quality Standard (AQS) for sulphur dioxide in the industrial East Montreal area. The SEC system is comprised of: 3 continuous SO{sub 2} monitoring stations with data loggers and remote communications; a meteorological tower with data logger and modem for acquiring local meteorology; communications with Environment Canada to download meteorological forecast data; a polling PC for data retrieval; and Windows NT basedmore » software running on the AIEM computer server. The SEC software utilizes relational databases to store and maintain measured SO{sub 2} concentration data, emission data, as well as observed and forecast meteorological data. The SEC system automatically executes a numerical dispersion model to forecast SO{sub 2} concentrations up to six hours in the future. Based on measured SO{sub 2} concentrations at the monitoring stations and the six hour forecast concentrations, the system determines if local sources should reduce their emission levels to avoid potential exceedences of the AQS. The SEC system also includes a Graphical User Interface (GUI) for user access to the system. The SEC system and software are described, and the accuracy of the system at forecasting SO{sub 2} concentrations is examined.« less

Protein secretion and membrane insertion systems in gram-negative bacteria.

PubMed

Saier, Milton H

2006-01-01

In contrast to other organisms, gram-negative bacteria have evolved numerous systems for protein export. Eight types are known that mediate export across or insertion into the cytoplasmic membrane, while eight specifically mediate export across or insertion into the outer membrane. Three of the former secretory pathway (SP) systems, type I SP (ISP, ABC), IIISP (Fla/Path) and IVSP (Conj/Vir), can export proteins across both membranes in a single energy-coupled step. A fourth generalized mechanism for exporting proteins across the two-membrane envelope in two distinct steps (which we here refer to as type II secretory pathways [IISP]) utilizes either the general secretory pathway (GSP or Sec) or the twin-arginine targeting translocase for translocation across the inner membrane, and either the main terminal branch or one of several protein-specific export systems for translocation across the outer membrane. We here survey the various well-characterized protein translocation systems found in living organisms and then focus on the systems present in gram-negative bacteria. Comparisons between these systems suggest specific biogenic, mechanistic and evolutionary similarities as well as major differences.

14 CFR Sec. 1-1 - Applicability of system of accounts and reports.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 14 Aeronautics and Space 4 2010-01-01 2010-01-01 false Applicability of system of accounts and reports. Sec. 1-1 Section 1-1 Aeronautics and Space OFFICE OF THE SECRETARY, DEPARTMENT OF TRANSPORTATION... AIR CARRIERS General Accounting Provisions Sec. 1-1 Applicability of system of accounts and reports...

14 CFR Sec. 1-1 - Applicability of system of accounts and reports.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 14 Aeronautics and Space 4 2011-01-01 2011-01-01 false Applicability of system of accounts and reports. Sec. 1-1 Section 1-1 Aeronautics and Space OFFICE OF THE SECRETARY, DEPARTMENT OF TRANSPORTATION... AIR CARRIERS General Accounting Provisions Sec. 1-1 Applicability of system of accounts and reports...

14 CFR Sec. 1-1 - Applicability of system of accounts and reports.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 14 Aeronautics and Space 4 2014-01-01 2014-01-01 false Applicability of system of accounts and reports. Sec. 1-1 Section 1-1 Aeronautics and Space OFFICE OF THE SECRETARY, DEPARTMENT OF TRANSPORTATION... AIR CARRIERS General Accounting Provisions Sec. 1-1 Applicability of system of accounts and reports...

14 CFR 1-1 - Applicability of system of accounts and reports.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 14 Aeronautics and Space 4 2012-01-01 2012-01-01 false Applicability of system of accounts and reports. Sec. 1-1 Section Sec. 1-1 Aeronautics and Space OFFICE OF THE SECRETARY, DEPARTMENT OF... CERTIFICATED AIR CARRIERS General Accounting Provisions Sec. 1-1 Applicability of system of accounts and...

14 CFR Sec. 1-1 - Applicability of system of accounts and reports.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 14 Aeronautics and Space 4 2013-01-01 2013-01-01 false Applicability of system of accounts and reports. Sec. 1-1 Section 1-1 Aeronautics and Space OFFICE OF THE SECRETARY, DEPARTMENT OF TRANSPORTATION... AIR CARRIERS General Accounting Provisions Sec. 1-1 Applicability of system of accounts and reports...

Flux agreement above a Scots pine plantation

NASA Astrophysics Data System (ADS)

Gay, L. W.; Vogt, R.; Bernhofer, Ch.; Blanford, J. H.

1996-03-01

The surface energy exchange of 12m high Scots pine plantation at Hartheim, Germany, was measured with a variety of methods during a 11-day period of fine weather in mid-May 1992. Net radiation and rate of thermal storage were measured with conventional net radiometers, soil heat flux discs and temperature-based storage models. The turbulent fluxes discussed in this report were obtained with an interchanging Bowen ratio energy budget system (BREB, at 14 m), two one-propeller eddy correlation systems (OPEC systems 1 and 2 at 17m), a 1-dimensional sonic eddy correlation system (SEC system 3) at 15 m, all on one “low” tower, and a 3-dimensional sonic eddy correlation system (SEC system 22) at 22 m on the “high” tower that was about 46 m distant. All systems measured sensible and latent heat (H and LE) directly, except for OPEC systems 1 and 2 which estimated LE as a residual term in the surface energy balance. Closure of turbulent fluxes from the two SEC systems was around 80% for daytime and 30% for night, with closure of 1-dimensional SEC system 3 exceeding that of 3-dimensional SEC system 22. The night measurements of turbulent fluxes contained considerable uncertainty, especially with the BREB system where measured gradients often yielded erroneous fluxes due to problems inherent in the method (i.e., computational instability as Bowen's ratio approaches -1). Also, both eddy correlation system designs (OPEC and SEC) appeared to underestimate |H| during stable conditions at night. In addition, both sonic systems (1- and 3-dimensional) underestimated |LE| during stable conditions. The underestimate of |H| at night generated residual estimates of OPEC LE containing a “phantom dew” error that erroneously decreased daily LE totals by about 10 percent. These special night problems are circumvented here by comparing results for daytime periods only, rather than for full days. To summarize, turbulent fluxes on the low tower from OPEC system 2 and the adjacent SEC system 3 were in reasonable agreement, while the BREB system appeared to overestimate H and underestimate LE; H and LE measured by SEC system 22 on the high tower were lower than from OPEC and SEC3 on the low tower. The turbulent flux measurements tended to converge, but the data exhibit unexplained differences between days, between systems, and between locations.

Discovery of Salmonella Virulence Factors Translocated via Outer Membrane Vesicles to Murine Macrophages.

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

Yoon, Hyunjin; Ansong, Charles; Adkins, Joshua N.

We have previously shown that the regulators SpvR, FruR, IHF, PhoP/PhoQ, SsrA/SsrB, SlyA, Hnr, RpoE, SmpB, CsrA, RpoS, Crp, OmpR/EnvZ, and Hfq are essential for Salmonella Typhimurium virulence in mice. Here we use quantitative LC-MS-based proteomics profiling of in-frame deletion mutants of these 14 regulators to identify proteins that are coordinately regulated by these virulence regulators and are thus presumably novel factors contributing to Salmonella pathogenesis. Putative candidate proteins from proteomics analysis were determined, which exhibited similar abundance profiles to those of Salmonella pathogenicity island (SPI)-2 type III secretion system (TTSS) proteins. A subset of 5 proteins including STM0082, STM1548,more » PdgL, STM1633, and STM3595 was selected for further analysis. All 5 proteins were expressed inside macrophage cells and STM0082 (SrfN) was secreted into host cytoplasm. Furthermore, deletion of STM0082 attenuated virulence in mice when administered intraperitoneally as determined by competitive index. srfN transcription was positively regulated by SsrAB, however, secretion was independent of SPI-2 TTSS as well as SPI-1 TTSS and flagella. Proteins including PagK and STM2585A, which are positively regulated by PhoP/PhoQ, have sec signal peptides as predicted for SrfN and were secreted into macrophage cytoplasm regardless of SPI-2 TTSS. Isolation of outer membrane vesicles (OMVs) revealed the presence of SrfN, PagK, and STM2585A inside vesicle compartments. This result is the first case showing delivery of virulence effectors via OMVs in S. Typhimurium. Moreover, Hfq regulation of SrfN translation suggests that small non-coding RNAs may be responsible for regulating effector protein expression.« less

Direct identification of the site of binding on the chaperone SecB for the amino terminus of the translocon motor SecA.

PubMed

Randall, Linda L; Henzl, Michael T

2010-06-01

Protein export mediated by the general secretory Sec system in Escherichia coli proceeds by a dynamic transfer of a precursor polypeptide from the chaperone SecB to the SecA ATPase motor of the translocon and subsequently into and through the channel of the membrane-embedded SecYEG heterotrimer. The complex between SecA and SecB is stabilized by several separate sites of contact. Here we have demonstrated directly an interaction between the N-terminal residues 2 through 11 of SecA and the C-terminal 13 residues of SecB by isothermal titration calorimetry and analytical sedimentation velocity centrifugation. We discuss the unusual binding properties of SecA and SecB in context of a model for transfer of the precursor along the pathway of export.

Components of the SNARE-containing regulon are co-regulated in root cells undergoing defense

PubMed Central

Klink, Vincent P.; Sharma, Keshav; Pant, Shankar R.; McNeece, Brant; Niraula, Prakash; Lawrence, Gary W.

2017-01-01

ABSTRACT The term regulon has been coined in the genetic model plant Arabidopsis thaliana, denoting a structural and physiological defense apparatus defined genetically through the identification of the penetration (pen) mutants. The regulon is composed partially by the soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptor (SNARE) syntaxin PEN1. PEN1 has homology to a Saccharomyces cerevisae gene that regulates a Secretion (Sec) protein, Suppressor of Sec 1 (Sso1p). The regulon is also composed of the β-glucosidase (PEN2) and an ATP binding cassette (ABC) transporter (PEN3). While important in inhibiting pathogen infection, limited observations have been made regarding the transcriptional regulation of regulon genes until now. Experiments made using the model agricultural Glycine max (soybean) have identified co-regulated gene expression of regulon components. The results explain the observation of hundreds of genes expressed specifically in the root cells undergoing the natural process of defense. Data regarding additional G. max genes functioning within the context of the regulon are presented here, including Sec 14, Sec 4 and Sec 23. Other examined G. max homologs of membrane fusion genes include an endosomal bromo domain-containing protein1 (Bro1), syntaxin6 (SYP6), SYP131, SYP71, SYP8, Bet1, coatomer epsilon (ε-COP), a coatomer zeta (ζ-COP) paralog and an ER to Golgi component (ERGIC) protein. Furthermore, the effectiveness of biochemical pathways that would function within the context of the regulon ave been examined, including xyloglucan xylosyltransferase (XXT), reticuline oxidase (RO) and galactinol synthase (GS). The experiments have unveiled the importance of the regulon during defense in the root and show how the deposition of callose relates to the process. PMID:28010187

Role of salivary epithelial toll-like receptors 2 and 4 in modulating innate immune responses in chronic periodontitis.

PubMed

Swaminathan, V; Prakasam, S; Puri, V; Srinivasan, M

2013-12-01

Chronic periodontitis is initiated by sequential colonization with a broad array of bacteria and is perpetuated by an immune-inflammatory response to the changing biofilm. Host recognition of microbes is largely mediated by toll-like receptors (TLRs), which interact with conserved pathogen-associated molecular patterns. Based on ligand recognition, TLR-2 and TLR-4 interact with most periodontal pathogens. Extracrevicular bacterial reservoirs, such as the oral epithelial cells, contribute to the persistence of periodontitis. Human saliva is a rich source of oral epithelial cells that express functional TLRs. In this study we investigated the role of salivary epithelial cell (SEC) TLR-2 and TLR-4 in patients with generalized chronic periodontitis. Unstimulated whole saliva (UWS) was collected from patients with generalized chronic periodontitis and from healthy individuals after obtaining informed consent. Epithelial cells isolated from each UWS sample were assessed for TLR-2, TLR-4, peptidoglycan recognition protein (PGRP)-3 and PGRP-4 by quantitative real-time PCR. In addition, the SECs were stimulated in vitro with microbial products for up to 24 h. The culture supernatant was assessed for cytokines by ELISA. Stimulation with TLR-2- or TLR-4-specific ligands induced cytokine secretion with differential kinetics and up-regulated TLR2 and TLR4 mRNAs, respectively, in cultures of SECs from patients with periodontitis. In addition, the SECs from patients with periodontitis exhibited reduced PGRP3 and PGRP4 mRNAs, the TLR-responsive genes with antibacterial properties. SECs derived from the UWS of patients with chronic periodontitis are phenotypically distinct and could represent potential resources for assessing the epithelial responses to periodontal pathogens in the course of disease progression and persistence. © 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Effects of Trx2p and Sec23p expression on stable production of hepatitis B surface antigen S domain in recombinant Saccharomyces cerevisiae.

PubMed

Park, Young-Kyoung; Jung, Sang-Min; Lim, Hyung-Kwon; Son, Young-Jin; Park, Yong-Cheol; Seo, Jin-Ho

2012-08-31

The S domain of hepatitis B virus surface antigen (sHBsAg) is the primary component for vaccine development against virus infection. For stable expression of sHBsAg in recombinant Saccharomyces cerevisiae, new accessory genes necessary for foreign protein expression were screened by DNA microarray. Among 600 genes of interest, genes coding for an activating protein of ATPase in Hsp90 (Aha1p), S. cerevisiae DnaJ (Scj1p), thioredoxin 2 (Trx2p) and a GTPase-activator specific for Sar1 (Sec23p) as well as Pdi1p were selected in transcriptome analysis, which are known to facilitate disulfide bond formation or induce protein transport in the secretion pathway. Individual and combinatorial expression of SEC23, TRX2 and PDI1 increased total sHBsAg concentration by 1.9-6.5-fold, relative to the control strain expressing sHBsAg only. Additionally, moderate expression of Kex2p protease able to cut off the signal peptide enhanced the portion of the authentic sHBsAg to total sHBsAg. Fed-batch fermentation of the S. cerevisiae 2805 strain coexpressing the sHBsAg, SEC23, PDI1 and KEX2 genes resulted in 70.6mg/L final sHBsAg concentration which was 5.6 times higher than that of the control. Transmission electron microscopic analysis of the yeast cells elucidated the effects of the accessory gene coexpression on the intracellular localization of sHBsAg. Like PDI1, coexpression of both SEC23 and/or TRX2 newly isolated in this study is expected to improve the target protein expression in S. cerevisiae. Copyright © 2012 Elsevier B.V. All rights reserved.

Decreased proteinase A excretion by strengthening its vacuolar sorting and weakening its constitutive secretion in Saccharomyces cerevisiae.

PubMed

Chen, Yefu; Song, Lulu; Han, Yueran; Liu, Mingming; Gong, Rui; Luo, Weiwei; Guo, Xuewu; Xiao, Dongguang

2017-01-01

Proteinase A (PrA), encoded by PEP4 gene, is detrimental to beer foam stability. There are two transport pathways for the new synthesized PrA in yeast, sorting to the vacuole normally, or excreting out of the cells under stress conditions. They were designated as the Golgi-to-vacuole pathway and the constitutive secretory pathway, respectively. To reduce PrA excretion in some new way instead of its coding gene deletion, which had a negative effect on cell metabolism and beer fermentation, we modified the PrA transport based on these above two pathways. In the Golgi-to-vacuole pathway, after the verification that Vps10p is the dominant sorting receptor for PrA Golgi-to-vacuolar transportation by VPS10 deletion, VPS10 was then overexpressed. Furthermore, SEC5, encoding exocyst complexes' central subunit (Sec5p) in the constitutive secretory pathway, was deleted. The results show that PrA activity in the broth fermented with WGV10 (VPS10 overexpressing strain) and W∆SEC5 (SEC5 deletion strain) was lowered by 76.96 and 32.39%, compared with the parental strain W303-1A, at the end of main fermentation. There are negligible changes in fermentation performance between W∆SEC5 and W303-1A, whereas, surprisingly, WGV10 had a significantly improved fermentation performance compared with W303-1A. WGV10 has an increased growth rate, resulting in higher biomass and faster fermentation speed; finally, wort fermentation is performed thoroughly. The results show that the biomass production of WGV10 is always higher than that of W∆SEC5 and W303-1A at all stages of fermentation, and that ethanol production of WGV10 is 1.41-fold higher than that of W303-1A. Obviously, VPS10 overexpression is beneficial for yeast and is a more promising method for reduction of PrA excretion.

Comparative Transcriptome Analysis of Vibrio splendidus JZ6 Reveals the Mechanism of Its Pathogenicity at Low Temperatures

PubMed Central

Liu, Rui; Chen, Hao; Zhang, Ran; Zhou, Zhi; Hou, Zhanhui; Gao, Dahai; Zhang, Huan; Wang, Lingling

2016-01-01

Yesso scallop-pathogenic Vibrio splendidus strain JZ6 was found to have the highest virulence at 10°C, while its pathogenicity was significantly reduced with increased temperature and completely incapacitated at 28°C. In the present study, comparative transcriptome analyses of JZ6 and another nonpathogenic V. splendidus strain, TZ19, were conducted at two crucial culture temperatures (10°C and 28°C) in order to determine the possible mechanism of temperature regulation of virulence. Comparisons among four libraries, constructed from JZ6 and TZ19 cultured at 10°C and 28°C (designated JZ6_10, JZ6_28, TZ19_10, and TZ19_28), revealed that 241 genes were possibly related to the increased virulence of JZ6 at 10°C. There were 10 genes, including 2 encoding Flp pilus assembly proteins (FlhG and VS_2437), 6 encoding proteins of the “Vibrio cholerae pathogenic cycle” (ToxS, CqsA, CqsS, RpoS, HapR, and Vsm), and 2 encoding proteins in the Sec-dependent pathway (SecE and FtsY), that were significantly upregulated in JZ6_10 (P < 0.05) compared to those in JZ6_28, TZ19_10, and TZ19_28, which were supposed to be responsible for adhesion, quorum sensing, virulence, and protein secretion of V. splendidus. When cultured at 10°C, JZ6 cells were larger and tended to aggregate more than those cultured at 28°C. The virulence factor (extracellular metalloprotease) was also found to be highly expressed in the extracellular product (ECP) of JZ6 at 10°C, and this ECP exhibited obvious cytotoxicity to oyster primary hemocytes, A549 cells, and L929 cells. These results indicated that low temperatures (10°C) could enhance adhesion, activate the quorum sensing systems, upregulate virulence factor synthesis and secretion, and, lastly, increase the pathogenicity of JZ6. PMID:26801576

An Intracellular Peptidyl-Prolyl cis/trans Isomerase Is Required for Folding and Activity of the Staphylococcus aureus Secreted Virulence Factor Nuclease.

PubMed

Wiemels, Richard E; Cech, Stephanie M; Meyer, Nikki M; Burke, Caleb A; Weiss, Andy; Parks, Anastacia R; Shaw, Lindsey N; Carroll, Ronan K

2017-01-01

Staphylococcus aureus is an important human pathogen that relies on a large repertoire of secreted and cell wall-associated proteins for pathogenesis. Consequently, the ability of the organism to cause disease is absolutely dependent on its ability to synthesize and successfully secrete these proteins. In this study, we investigate the role of peptidyl-prolyl cis/trans isomerases (PPIases) on the activity of the S. aureus secreted virulence factor nuclease (Nuc). We identify a staphylococcal cyclophilin-type PPIase (PpiB) that is required for optimal activity of Nuc. Disruption of ppiB results in decreased nuclease activity in culture supernatants; however, the levels of Nuc protein are not altered, suggesting that the decrease in activity results from misfolding of Nuc in the absence of PpiB. We go on to demonstrate that PpiB exhibits PPIase activity in vitro, is localized to the bacterial cytosol, and directly interacts with Nuc in vitro to accelerate the rate of Nuc refolding. Finally, we demonstrate an additional role for PpiB in S. aureus hemolysis and demonstrate that the S. aureus parvulin-type PPIase PrsA also plays a role in the activity of secreted virulence factors. The deletion of prsA leads to a decrease in secreted protease and phospholipase activity, similar to that observed in other Gram-positive pathogens. Together, these results demonstrate, for the first time to our knowledge, that PPIases play an important role in the secretion of virulence factors in S. aureus IMPORTANCE: Staphylococcus aureus is a highly dangerous bacterial pathogen capable of causing a variety of infections throughout the human body. The ability of S. aureus to cause disease is largely due to an extensive repertoire of secreted and cell wall-associated proteins, including adhesins, toxins, exoenzymes, and superantigens. These virulence factors, once produced, are typically transported across the cell membrane by the secretory (Sec) system in a denatured state. Consequently, once outside the cell, they must refold into their active form. This step often requires the assistance of bacterial folding proteins, such as PPIases. In this work, we investigate the role of PPIases in S. aureus and uncover a cyclophilin-type enzyme that assists in the folding/refolding of staphylococcal nuclease. Copyright © 2016 American Society for Microbiology.

Prediction of selenoprotein T structure and its response to selenium deficiency in chicken immune organs.

PubMed

You, Lu; Liu, Ci; Yang, Zi-Jiang; Li, Ming; Li, Shu

2014-08-01

Selenoprotein T (SelT) is associated with the regulation of calcium homeostasis and neuroendocrine secretion. SelT can also change cell adhesion and is involved in redox regulation and cell fixation. However, the structure and function of chicken SelT and its response to selenium (Se) remains unclear. In the present study, 150 1-day-old chickens were randomly divided into a low Se group (L group, fed a Se-deficient diet containing 0.020 mg/kg Se) and a control group (C group, fed a diet containing sodium selenite at 0.2 mg/kg Se). The immune organs (spleen, thymus, and bursa of Fabricius) were collected at 15, 25, 35, 45, and 55 days of age. We performed a sequence analysis and predicted the structure and function of SelT. We also investigated the effects of Se deficiency on the expression of SelT, selenophosphate synthetase-1 (SPS1), and selenocysteine synthase (SecS) using RT-PCR and the oxidative stress in the chicken immune organs. The data showed that the coding sequence (CDS) and deduced amino acid sequence of SelT were highly similar to those of 17 other animals. Se deficiency induced lower (P < 0.05) levels of SelT, SPS1, and SecS, reduced the catalase (CAT) activity, and increased the levels of hydrogen peroxide (H2O2) and hydroxyl radical (-OH) in immune organs. In conclusion, the CDS and deduced amino acid sequence of chicken SelT are highly homologous to those of various mammals. The redox function and response to the Se deficiency of chicken SelT may be conserved. A Se-deficient diet led to a decrease in SelT, SecS, and SPS1 and induced oxidative stress in the chicken immune organs. To our knowledge, this is the first report of predictions of chicken SelT structure and function. The present study demonstrated the relationship between the selenoprotein synthases (SPS1, SecS) and SelT expression in the chicken immune organs and further confirmed oxidative stress caused by Se deficiency. Thus, the information presented in this study is helpful to understand chicken SelT structure and function. Meanwhile, the present research also confirmed the negative effects of Se deficiency on chicken immune organs.

Information Systems: Opportunities Exist to Strengthen SEC’s Oversight of Capacity and Security

DTIC Science & Technology

2001-07-01

Strengthen SEC’s Oversight of Capacity and Security 5 . FUNDING NUMBERS 6. AUTHOR(S) GAO 7. PERFORMING ORGANIZATION NAME(S) AND ADDRESS(ES) 8. PERFORMING...ANSI Std. Z39-18 298-102 Page i GAO-01-863 Information Systems Letter 1 Results in Brief 2 Background 4 Scope and Methodology 5 SEC Uses a Wide Range...or external organizations to conduct the independent reviews. These internal audits are performed cyclically based on an annual risk analysis. SEC

The importance of system band broadening in modern size exclusion chromatography.

PubMed

Goyon, Alexandre; Guillarme, Davy; Fekete, Szabolcs

2017-02-20

In the last few years, highly efficient UHP-SEC columns packed with sub-3μm particles were commercialized by several providers. Besides the particle size reduction, the dimensions of modern SEC stationary phases (150×4.6mm) was also modified compared to regular SEC columns (300×6 or 300×8mm). Because the analytes are excluded from the pores in SEC, the retention factors are very low, ranging from -1

Communication satellite payload technologies - State of the art and trends in Europe

NASA Astrophysics Data System (ADS)

Mica, G.; Coirault, R.

1982-09-01

Communication satellite payload technologies are examined, in terms of past, present, and future ESA guidelines. Various existing payload systems are presented, such as Marecs, ECS, and L-Sat (which will carry four payloads). Future services within the market include 2 Mb/sec high speed data, 2-8 Mb/sec video conference, and 64 Mb/sec television distribution, and growth in these areas is dependent on traffic requirements. Pre-operational satellites are outlined, for example Telecom 1 has an estimated system capacity of 150 Mb/sec, Italsat has an expected 1180 Mb/sec, and DFS demonstrates a possible 1540 Mb/sec capacity. It is found that the 20/30 GHz band should be applied for use in wideband and high capacity trunks among heavy traffic centers. To accommodate for the noise in this waveband, the parametric amplifier developed for L-Sat must be used. Finally, development objectives for future programs include improving spectrum and geostationary orbit utilization, cost-efficiency, and standardization of systems.

Knocking out Bcsas1 in Botrytis cinerea impacts growth, development, and secretion of extracellular proteins, which decreases virulence.

PubMed

Zhang, Zhanquan; Qin, Guozheng; Li, Boqiang; Tian, Shiping

2014-06-01

Pathogenic fungi usually secrete a series of virulence factors to the extracellular environment to facilitate infection. Rab GTPases play a central role in the secretory pathway. To explore the function of Rab/GTPase in filamentous fungi, we knocked out a Rab/GTPase family gene, Bcsas1, in Botrytis cinerea, an aggressive fungal pathogen that infects more than 200 plant species. A detailed analysis was conducted on the virulence and the secretory capability of the mutants. The results indicated that knockout of Bcsas1 inhibited hyphal development and reduced sporulation of B. cinerea on potato dextrose agar plates resulting in reduced virulence on various fruit hosts. Knocking out the Bcsas1 gene led to an accumulation of transport vesicles at the hyphal tip, significantly reduced extracellular protein content, and lowered the activity of polygalacturonase and xylanase in the extracellular medium. However, mutation of Bcsas1 did not affect the expression of genes encoding polygalacturonase and xylanase, suggesting the secretion of these two family enzymes was suppressed in the mutant. Moreover, a comparative analysis of the secretome provided further evidence that the disruption of Bcsas1 in mutant strains significantly depressed the secretion of polysaccharide hydrolases and proteases. The results indicate that Bcsas1, the Rab8/SEC4-like gene, plays a crucial role in development, protein secretion, and virulence of B. cinerea.

What Makes a Bacterial Species Pathogenic?:Comparative Genomic Analysis of the Genus Leptospira

PubMed Central

Fouts, Derrick E.; Matthias, Michael A.; Adhikarla, Haritha; Adler, Ben; Amorim-Santos, Luciane; Berg, Douglas E.; Bulach, Dieter; Buschiazzo, Alejandro; Chang, Yung-Fu; Galloway, Renee L.; Haake, David A.; Haft, Daniel H.; Hartskeerl, Rudy; Ko, Albert I.; Levett, Paul N.; Matsunaga, James; Mechaly, Ariel E.; Monk, Jonathan M.; Nascimento, Ana L. T.; Nelson, Karen E.; Palsson, Bernhard; Peacock, Sharon J.; Picardeau, Mathieu; Ricaldi, Jessica N.; Thaipandungpanit, Janjira; Wunder, Elsio A.; Yang, X. Frank; Zhang, Jun-Jie; Vinetz, Joseph M.

2016-01-01

Leptospirosis, caused by spirochetes of the genus Leptospira, is a globally widespread, neglected and emerging zoonotic disease. While whole genome analysis of individual pathogenic, intermediately pathogenic and saprophytic Leptospira species has been reported, comprehensive cross-species genomic comparison of all known species of infectious and non-infectious Leptospira, with the goal of identifying genes related to pathogenesis and mammalian host adaptation, remains a key gap in the field. Infectious Leptospira, comprised of pathogenic and intermediately pathogenic Leptospira, evolutionarily diverged from non-infectious, saprophytic Leptospira, as demonstrated by the following computational biology analyses: 1) the definitive taxonomy and evolutionary relatedness among all known Leptospira species; 2) genomically-predicted metabolic reconstructions that indicate novel adaptation of infectious Leptospira to mammals, including sialic acid biosynthesis, pathogen-specific porphyrin metabolism and the first-time demonstration of cobalamin (B12) autotrophy as a bacterial virulence factor; 3) CRISPR/Cas systems demonstrated only to be present in pathogenic Leptospira, suggesting a potential mechanism for this clade’s refractoriness to gene targeting; 4) finding Leptospira pathogen-specific specialized protein secretion systems; 5) novel virulence-related genes/gene families such as the Virulence Modifying (VM) (PF07598 paralogs) proteins and pathogen-specific adhesins; 6) discovery of novel, pathogen-specific protein modification and secretion mechanisms including unique lipoprotein signal peptide motifs, Sec-independent twin arginine protein secretion motifs, and the absence of certain canonical signal recognition particle proteins from all Leptospira; and 7) and demonstration of infectious Leptospira-specific signal-responsive gene expression, motility and chemotaxis systems. By identifying large scale changes in infectious (pathogenic and intermediately pathogenic) vs. non-infectious Leptospira, this work provides new insights into the evolution of a genus of bacterial pathogens. This work will be a comprehensive roadmap for understanding leptospirosis pathogenesis. More generally, it provides new insights into mechanisms by which bacterial pathogens adapt to mammalian hosts. PMID:26890609

What Makes a Bacterial Species Pathogenic?:Comparative Genomic Analysis of the Genus Leptospira.

PubMed

Fouts, Derrick E; Matthias, Michael A; Adhikarla, Haritha; Adler, Ben; Amorim-Santos, Luciane; Berg, Douglas E; Bulach, Dieter; Buschiazzo, Alejandro; Chang, Yung-Fu; Galloway, Renee L; Haake, David A; Haft, Daniel H; Hartskeerl, Rudy; Ko, Albert I; Levett, Paul N; Matsunaga, James; Mechaly, Ariel E; Monk, Jonathan M; Nascimento, Ana L T; Nelson, Karen E; Palsson, Bernhard; Peacock, Sharon J; Picardeau, Mathieu; Ricaldi, Jessica N; Thaipandungpanit, Janjira; Wunder, Elsio A; Yang, X Frank; Zhang, Jun-Jie; Vinetz, Joseph M

2016-02-01

Leptospirosis, caused by spirochetes of the genus Leptospira, is a globally widespread, neglected and emerging zoonotic disease. While whole genome analysis of individual pathogenic, intermediately pathogenic and saprophytic Leptospira species has been reported, comprehensive cross-species genomic comparison of all known species of infectious and non-infectious Leptospira, with the goal of identifying genes related to pathogenesis and mammalian host adaptation, remains a key gap in the field. Infectious Leptospira, comprised of pathogenic and intermediately pathogenic Leptospira, evolutionarily diverged from non-infectious, saprophytic Leptospira, as demonstrated by the following computational biology analyses: 1) the definitive taxonomy and evolutionary relatedness among all known Leptospira species; 2) genomically-predicted metabolic reconstructions that indicate novel adaptation of infectious Leptospira to mammals, including sialic acid biosynthesis, pathogen-specific porphyrin metabolism and the first-time demonstration of cobalamin (B12) autotrophy as a bacterial virulence factor; 3) CRISPR/Cas systems demonstrated only to be present in pathogenic Leptospira, suggesting a potential mechanism for this clade's refractoriness to gene targeting; 4) finding Leptospira pathogen-specific specialized protein secretion systems; 5) novel virulence-related genes/gene families such as the Virulence Modifying (VM) (PF07598 paralogs) proteins and pathogen-specific adhesins; 6) discovery of novel, pathogen-specific protein modification and secretion mechanisms including unique lipoprotein signal peptide motifs, Sec-independent twin arginine protein secretion motifs, and the absence of certain canonical signal recognition particle proteins from all Leptospira; and 7) and demonstration of infectious Leptospira-specific signal-responsive gene expression, motility and chemotaxis systems. By identifying large scale changes in infectious (pathogenic and intermediately pathogenic) vs. non-infectious Leptospira, this work provides new insights into the evolution of a genus of bacterial pathogens. This work will be a comprehensive roadmap for understanding leptospirosis pathogenesis. More generally, it provides new insights into mechanisms by which bacterial pathogens adapt to mammalian hosts.

Suppressor Analysis Reveals a Role for SecY in the SecA2-Dependent Protein Export Pathway of Mycobacteria

PubMed Central

Ligon, Lauren S.; Rigel, Nathan W.; Romanchuk, Artur; Jones, Corbin D.

2013-01-01

All bacteria use the conserved Sec pathway to transport proteins across the cytoplasmic membrane, with the SecA ATPase playing a central role in the process. Mycobacteria are part of a small group of bacteria that have two SecA proteins: the canonical SecA (SecA1) and a second, specialized SecA (SecA2). The SecA2-dependent pathway exports a small subset of proteins and is required for Mycobacterium tuberculosis virulence. The mechanism by which SecA2 drives export of proteins across the cytoplasmic membrane remains poorly understood. Here we performed suppressor analysis on a dominant negative secA2 mutant (secA2 K129R) of the model mycobacterium Mycobacterium smegmatis to better understand the pathway used by SecA2 to export proteins. Two extragenic suppressor mutations were identified as mapping to the promoter region of secY, which encodes the central component of the canonical Sec export channel. These suppressor mutations increased secY expression, and this effect was sufficient to alleviate the secA2 K129R phenotype. We also discovered that the level of SecY protein was greatly diminished in the secA2 K129R mutant, but at least partially restored in the suppressors. Furthermore, the level of SecY in a suppressor strongly correlated with the degree of suppression. Our findings reveal a detrimental effect of SecA2 K129R on SecY, arguing for an integrated system in which SecA2 works with SecY and the canonical Sec translocase to export proteins. PMID:23913320

Implementation of the Single European Code in a Multi-Tissue Bank.

PubMed

Schroeter, Jan; Schulz, Tino; Schroeter, Bernard; Fleischhauer, Katrin; Pruß, Axel

2017-11-01

The traceability of tissue and cells transplants is important to ensure a high level of safety for the recipients. With the final introduction of the Single European Code (SEC) in April 2017 in the EU a consistent system among all member states became mandatory. The regulations for the SEC on EU and national level were evaluated. An overview on the different parts of the SEC with detailed explanations is given. Our own experiences with the implementation of the SEC in our multi-tissue bank are reported in addition. The implementation of the SEC in our multi-tissue bank could be successfully realized. However, it revealed a number of difficulties, especially the sterile labeling of certain tissue transplants and the complex update of the existing database. The introduction of the SEC has made a contribution to the safety of recipients of tissue and cells transplants through a system of comprehensive and transparent traceability.

Sec13 safeguards the integrity of the endoplasmic reticulum and organogenesis of the digestive system in zebrafish.

PubMed

Niu, Xubo; Gao, Chuan; Jan Lo, Li; Luo, Yue; Meng, Chunmei; Hong, Jian; Hong, Wanjin; Peng, Jinrong

2012-07-15

The Sec13-Sec31 heterotetramer serves as the outer coat in the COPII complex, which mediates protein trafficking from the endoplasmic reticulum (ER) to the Golgi apparatus. Although it has been studied in depth in yeast and cultured cells, the role of COPII in organogenesis in a multicellular organism has not. We report here that a zebrafish sec13(sq198) mutant, which exhibits a phenotype of hypoplastic digestive organs, has a mutation in the sec13 gene. The mutant gene encodes a carboxyl-terminus-truncated Sec13 that loses its affinity to Sec31a, which leads to disintegration of the ER structure in various differentiated cells in sec13(sq198), including chondrocytes, intestinal epithelial cells and hepatocytes. Disruption of the ER structure activates an unfolded protein response that eventually causes the cells to undergo cell-cycle arrest and cell apoptosis, which arrest the growth of developing digestive organs in the mutant. Our data provide the first direct genetic evidence that COPII function is essential for the organogenesis of the digestive system. Copyright © 2012 Elsevier Inc. All rights reserved.

Exotoxin A of Pseudomonas aeruginosa: active, cloned toxin is secreted into the periplasmic space of Escherichia coli.

PubMed

Douglas, C M; Guidi-Rontani, C; Collier, R J

1987-11-01

We subcloned the structural gene for exotoxin A (ETA) of Pseudomonas aeruginosa in front of the tac promoter in an Escherichia coli expression vector and studied the intracellular location and properties of the protein product. The E. coli K-12 strain that carried this recombinant plasmid produced an immunoreactive protein that was identical to authentic ETA in size and in cytotoxic and ADP-ribosyl transferase activities per unit of immunoreactive material. The protein was predominantly in the periplasmic fraction; and a mutation in the secA gene blocked secretion, processing, and conversion of the protein to a fully toxic conformation. The results indicate that expression of the ETA gene in E. coli yields native ETA, which is localized within the periplasmic space. This organism may therefore serve as a useful host for studying structure and function in ETA.

Implementation of the Single European Code in a Multi-Tissue Bank

PubMed Central

Schroeter, Jan; Schulz, Tino; Schroeter, Bernard; Fleischhauer, Katrin; Pruß, Axel

2017-01-01

Introduction The traceability of tissue and cells transplants is important to ensure a high level of safety for the recipients. With the final introduction of the Single European Code (SEC) in April 2017 in the EU a consistent system among all member states became mandatory. Methods The regulations for the SEC on EU and national level were evaluated. An overview on the different parts of the SEC with detailed explanations is given. Our own experiences with the implementation of the SEC in our multi-tissue bank are reported in addition. Results The implementation of the SEC in our multi-tissue bank could be successfully realized. However, it revealed a number of difficulties, especially the sterile labeling of certain tissue transplants and the complex update of the existing database. Conclusion The introduction of the SEC has made a contribution to the safety of recipients of tissue and cells transplants through a system of comprehensive and transparent traceability. PMID:29344015

SEC sensor parametric test and evaluation system

NASA Technical Reports Server (NTRS)

1978-01-01

This system provides the necessary automated hardware required to carry out, in conjunction with the existing 70 mm SEC television camera, the sensor evaluation tests which are described in detail. The Parametric Test Set (PTS) was completed and is used in a semiautomatic data acquisition and control mode to test the development of the 70 mm SEC sensor, WX 32193. Data analysis of raw data is performed on the Princeton IBM 360-91 computer.

User’s Manual for the Generalized Computer Program System Open-Channel Flow and Sedimentation TABS-2. Main Text and Appendices A through O.

DTIC Science & Technology

1985-08-01

American Society of Civil Engineers,, No. HYIL. Ariathurai, R. 1974 (Aug). A Finite Element Model for Sediment Transport in Estuaries, Ph. D. Thesis ...11 aml ms po GR PH C 1.2 ś, I i 2 "’-..04/85 a I -- . a.., 4 4 - 4 I --.. - £ . ,. 04/85 4 444 4 9*4*4449 444 A4 A44 4 4 4 4 4 4 * 4 44 4 4 4 p 4...VEL MAGNITUDE (HOURS) CFT /SEC) (FT/SEC) (F/SEC) (FT) (FT) STATUS (FiSEC) (FT/SEC) (FT/SEC) .00 -. 115 -. 043 -. 123 5.501 100.507 MET -. 112 -. 069

76 FR 55293 - Special Conditions: Diamond Aircraft Industries, Model DA-40NG; Electronic Engine Control (EEC...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-09-07

... prior to the codification of Sec. 23.1308. There are several difficulties for propulsion systems... for Sec. 23.1309 and propulsion system capabilities and failure susceptibilities. The following figure...

14 CFR Sec. 2-4 - Accounting period.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 14 Aeronautics and Space 4 2010-01-01 2010-01-01 false Accounting period. Sec. 2-4 Section 2-4... REGULATIONS UNIFORM SYSTEM OF ACCOUNTS AND REPORTS FOR LARGE CERTIFICATED AIR CARRIERS General Accounting Provisions Sec. 2-4 Accounting period. (a) The accounting year of each air carrier subject to this Uniform...

14 CFR Sec. 1-6 - Accounting entities.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 14 Aeronautics and Space 4 2010-01-01 2010-01-01 false Accounting entities. Sec. 1-6 Section 1-6... REGULATIONS UNIFORM SYSTEM OF ACCOUNTS AND REPORTS FOR LARGE CERTIFICATED AIR CARRIERS General Accounting Provisions Sec. 1-6 Accounting entities. (a) Separate accounting records shall be maintained for each air...

Experiments of 10 Gbit/sec quantum stream cipher applicable to optical Ethernet and optical satellite link

NASA Astrophysics Data System (ADS)

Hirota, Osamu; Ohhata, Kenichi; Honda, Makoto; Akutsu, Shigeto; Doi, Yoshifumi; Harasawa, Katsuyoshi; Yamashita, Kiichi

2009-08-01

The security issue for the next generation optical network which realizes Cloud Computing System Service with data center" is urgent problem. In such a network, the encryption by physical layer which provide super security and small delay should be employed. It must provide, however, very high speed encryption because the basic link is operated at 2.5 Gbit/sec or 10 Gbit/sec. The quantum stream cipher by Yuen-2000 protocol (Y-00) is a completely new type random cipher so called Gauss-Yuen random cipher, which can break the Shannon limit for the symmetric key cipher. We develop such a cipher which has good balance of the security, speed and cost performance. In SPIE conference on quantum communication and quantum imaging V, we reported a demonstration of 2.5 Gbit/sec system for the commercial link and proposed how to improve it to 10 Gbit/sec. This paper reports a demonstration of the Y-00 cipher system which works at 10 Gbit/sec. A transmission test in a laboratory is tried to get the basic data on what parameters are important to operate in the real commercial networks. In addition, we give some theoretical results on the security. It is clarified that the necessary condition to break the Shannon limit requires indeed the quantum phenomenon, and that the full information theoretically secure system is available in the satellite link application.

Signal transduction in neurons: effects of cellular prion protein on fyn kinase and ERK1/2 kinase.

PubMed

Tomasi, Vittorio

2010-12-16

It has been reported that cellular prion protein (PrPc) co-localizes with caveolin-1 and participates to signal transduction events by recruiting Fyn kinase. As PrPc is a secreted protein anchored to the outer surface membrane through a glycosylphosphatidylinositol (GPI) anchor (secPrP) and caveolin-1 is located in the inner leaflet of plasma membrane, there is a problem of how the two proteins can physically interact each other and transduce signals. By using the GST-fusion proteins system we observed that PrPc strongly interacts with caveolin-1 scaffolding domain and with a caveolin-1 hydrophilic C-terminal region, but not with the caveolin-1 N-terminal region. In vitro binding experiments were also performed to define the site(s) of PrPc interacting with cav-1. The results are consistent with a participation of PrPc octapeptide repeats motif in the binding to caveolin-1 scaffolding domain. The caveolar localization of PrPc was ascertained by co-immunoprecipitation, by co-localization after flotation in density gradients and by confocal microscopy analysis of PrPc and caveolin-1 distributions in a neuronal cell line (GN11) expressing caveolin-1 at high levels. We observed that, after antibody-mediated cross-linking or copper treatment, PrPc was internalized probably into caveolae. We propose that following translocation from rafts to caveolae or caveolae-like domains, secPrP could interact with caveolin-1 and induce signal transduction events.

Identification and characterization of a translation arrest motif in VemP by systematic mutational analysis.

PubMed

Mori, Hiroyuki; Sakashita, Sohei; Ito, Jun; Ishii, Eiji; Akiyama, Yoshinori

2018-02-23

VemP ( V ibrio protein e xport m onitoring p olypeptide) is a secretory protein comprising 159 amino acid residues, which functions as a secretion monitor in Vibrio and regulates expression of the downstream V.secDF2 genes. When VemP export is compromised, its translation specifically undergoes elongation arrest at the position where the Gln 156 codon of vemP encounters the P-site in the translating ribosome, resulting in up-regulation of V.SecDF2 production. Although our previous study suggests that many residues in a highly conserved C-terminal 20-residue region of VemP contribute to its elongation arrest, the exact role of each residue remains unclear. Here, we constructed a reporter system to easily and exactly monitor the in vivo arrest efficiency of VemP. Using this reporter system, we systematically performed a mutational analysis of the 20 residues (His 138 -Phe 157 ) to identify and characterize the arrest motif. Our results show that 15 residues in the conserved region participate in elongation arrest and that multiple interactions between important residues in VemP and in the interior of the exit tunnel contribute to the elongation arrest of VemP. The arrangement of these important residues induced by specific secondary structures in the ribosomal tunnel is critical for the arrest. Pro scanning analysis of the preceding segment (Met 120 -Phe 137 ) revealed a minor role of this region in the arrest. Considering these results, we conclude that the arrest motif in VemP is mainly composed of the highly conserved multiple residues in the C-terminal region. © 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

77 FR 15143 - Privacy Act of 1974; System of Records

Federal Register 2010, 2011, 2012, 2013, 2014

2012-03-14

...); and Education Jobs Fund, Public Law 111-226, Sec. 101, 124 Stat. 2389 (2010). Accordingly, the Board... classification, system location, storage, retrievability, safeguards, retention and disposal, and system manager.... Sec. 1521, 1523(a)(1), 123 Stat. 115, 289-90 (2009) (Recovery Act), and Education Jobs Fund, Public...

Temperature distortion generator for turboshaft engine testing

NASA Technical Reports Server (NTRS)

Klann, G. A.; Barth, R. L.; Biesiadny, T. J.

1984-01-01

The procedures and unique hardware used to conduct an experimental investigation into the response of a small-turboshaft-engine compression system to various hot gas ingestion patterns are presented. The temperature distortion generator described herein uses gaseous hydrogen to create both steady-state and time-variant, or transient, temperature distortion at the engine inlet. The range of transient temperature ramps produced by the distortion generator during the engine tests was from less than 111 deg K/sec (200 deg R/sec) to above 611 deg K/sec (1100 deg R/sec); instantaneous temperatures to 422 deg K (760 deg R) above ambient were generated. The distortion generator was used to document the maximum inlet temperatures and temperature rise rates that the compression system could tolerate before the onset of stall for various circumferential distortions as well as the compressor system response during stall.

77 FR 5860 - Privacy Act of 1974; System of Records

Federal Register 2010, 2011, 2012, 2013, 2014

2012-02-06

..., 2011); Education Jobs Fund, Public Law 111-226, 124 Stat. 2389, Sec. 101 (2010). Accordingly, the Board... fraud, waste, and abuse. Public Law 111-5, Sec. Sec. 1521, 1523(a)(1). The Education Jobs Fund required... contains information relevant to the retention of an employee or retention of a security clearance. That...

Near-Infrared Fluorescent Turn-on Probe with a Remarkable Large Stokes Shift for Imaging Selenocysteine in Living Cells and Animals.

PubMed

Feng, Weiyong; Li, Meixing; Sun, Yao; Feng, Guoqiang

2017-06-06

Selenocysteine (Sec) is the 21st naturally occurring amino acid and has emerged as an important sensing target in recent years. However, fluorescent detection of Sec in living systems is challenging. To date, very few fluorescent Sec probes have been reported and most of them respond fluorescence to Sec in the visible region. In this paper, a very promising near-infrared fluorescent probe for Sec was developed. This probe works in aqueous solution over a wide pH range under mild conditions and can be used for rapid, highly selective and sensitive detection of Sec with significant near-infrared fluorescent turn-on signal changes. In addition, it features a remarkable large Stokes shift (192 nm) and a low detection limit (60 nM) for Sec with a wide linear range (0-70 μM). Moreover, this probe can be conveniently used to detect Sec in serum samples, living cells, and animals, indicating it holds great promise for biological applications.

Natural and Heterologous Production of Bacteriocins

NASA Astrophysics Data System (ADS)

Cintas, Luis M.; Herranz, Carmen; Hernández, Pablo E.

Bacteriocins are ribosomally synthesized antimicrobial peptides produced by bacteria, and their use as natural and nontoxic food preservatives has been the source of considerable interest for the research community. In addition, bacteriocins have been investigated for their potential use in human and veterinary applications and in the animal production field. In the native bacterial strain, most bacteriocins are synthesized as biologically inactive precursors, with N-terminal extensions, that are cleaved concomitantly during export of the bacteriocin by dedicated ABC transporters, or the general secretory pathway (GSP) or Sec-dependent pathway. However, a few bacteriocins are synthesized without an N-terminal extension, and others are circularized through a head-to-tail peptide bond, complicating the elucidation of their processing and transport across the cytoplasmic membrane. The high cost of synthetic bacteriocin synthesis and their low yields from many natural producers recommends the exploration of recombinant microbial systems for the heterologous production of bacteriocins. Other advantages of such systems include production of bacteriocins in safer hosts, increased bacteriocin production, control of bacteriocin gene expression, production of food ingredients with antimicrobial activity, construction of multibacteriocinogenic strains with a wider antagonistic spectrum, a better adaptation of the selected hosts to food environments, and providing antagonistic properties to lactic acid bacteria (LAB) used as starter, protective, or probiotic cultures. The recombinant production of bacteriocins mostly relies on the use of expression vectors that replicate in Gram-negative bacteria, Gram-positive bacteria, and yeasts, whereas the production of bacteriocins in heterologous LAB hosts may be essentially based on the expression of native biosynthetic genes, by exchanging or replacing leader peptides and/or dedicated processing and secretion systems (ABC transporters), or by fusion of mature bacteriocins to signal peptides that act as secretion signals.

Secretory expression of a heterologous protein, Aiio-AIO6BS, in Bacillus subtilis via a non-classical secretion pathway.

PubMed

Pan, Xingliang; Yang, Yalin; Liu, Xuewei; Li, Dong; Li, Juan; Guo, Xiaoze; Zhou, Zhigang

2016-09-16

The quenching enzyme AIO6 (AiiO-AIO6) has been reported as a feed additive preparation for application in aquaculture and biological control of pathogenic Aeromonas hydrophila. We developed an economical strategy to express AIO6BS (AiiO-AIO6BS, codon optimized AIO6 in Bacillus subtilis) in Bacillus subtilis for facilitating its widespread application. Promoter p43 without the signal peptide was used for secretory expression of AIO6BS in B. subtilis. Western blotting analysis demonstrated that AIO6BS was successfully expressed and secreted into the cell culture. Expression analysis of AIO6BS in the single or double mutant of the lytC and lytD genes for cell autolysis in B. subtilis 1A751 and cell autolysis-resistant engineered strain LM2531 derived from the wild type 168 indicated that the release of the heterologous protein AIO6BS was not simply mediated by cell lysis. Expression level of AIO6BS did not change in the mutants of B. subtilis that harbored mutations in the secA, tatAC, or ecsA genes compared with that in the parent wild type strain. These results suggested the AIO6BS protein was likely secreted via a non-classical secretion pathway. The expression analysis of the various N- or C-terminal truncated gene products indicated that AIO6BS probably acts as an export signal to direct its self-secretion across the cell membrane. Copyright © 2016 Elsevier Inc. All rights reserved.

20 CFR 662.230 - What are the responsibilities of the required One-Stop partners?

Code of Federal Regulations, 2010 CFR

2010-04-01

... delivery system the core services that are applicable to the partner's programs; (WIA sec. 121(b)(1)(A... delivery system; and (2) Provide core services; (WIA sec. 134(d)(1)(B).) (c) Enter into a memorandum of...

77 FR 69393 - Unique Device Identification System

Federal Register 2010, 2011, 2012, 2013, 2014

2012-11-19

... submitted to the GUDID data base. Sec. 830.300. Two years after publication of a final rule.... The label... are required to be labeled with a UDI, must be submitted to the GUDID data base. Sec. 830.300. All... a UDI, must be submitted to the GUDID data base. Sec. 830.300. Five years after publication of a...

Neutron radiographic viewing system

NASA Technical Reports Server (NTRS)

1972-01-01

The design, development and application of a neutron radiographic viewing system for use in nondestructive testing applications is considered. The system consists of a SEC vidicon camera, neutron image intensifier system, disc recorder, and TV readout. Neutron bombardment of the subject is recorded by an image converter and passed through an optical system into the SEC vidicon. The vidicon output may be stored, or processed for visual readout.

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

Kim, Moon-Jung; Lee, Byung Cheon; Division of Biotechnology, College of Life Sciences & Biotechnology, Korea University, Seoul 136-701

Thioredoxin (Trx) is a major thiol-disulfide reductase that plays a role in many biological processes, including DNA replication and redox signaling. Although selenocysteine (Sec)-containing Trxs have been identified in certain bacteria, their enzymatic properties have not been characterized. In this study, we expressed a selenoprotein Trx from Treponema denticola, an oral spirochete, in Escherichia coli and characterized this selenoenzyme and its natural cysteine (Cys) homologue using E. coli Trx1 as a positive control. {sup 75}Se metabolic labeling and mutation analyses showed that the SECIS (Sec insertion sequence) of T. denticola selenoprotein Trx is functional in the E. coli Sec insertion system with specificmore » selenium incorporation into the Sec residue. The selenoprotein Trx exhibited approximately 10-fold higher catalytic activity than the Sec-to-Cys version and natural Cys homologue and E. coli Trx1, suggesting that Sec confers higher catalytic activity on this thiol-disulfide reductase. Kinetic analysis also showed that the selenoprotein Trx had a 30-fold higher K{sub m} than Cys-containing homologues, suggesting that this selenoenzyme is adapted to work efficiently with high concentrations of substrate. Collectively, the results of this study support the hypothesis that selenium utilization in oxidoreductase systems is primarily due to the catalytic advantage provided by the rare amino acid, Sec. - Highlights: • The first characterization of a selenoprotein Trx is presented. • The selenoenzyme Trx exhibits 10-fold higher catalytic activity than Cys homologues. • Se utilization in Trx is primarily due to the catalytic advantage provided by Sec residue.« less

75 FR 9968 - Privacy Act of 1974: Systems of Records

Federal Register 2010, 2011, 2012, 2013, 2014

2010-03-04

... records: ``Mailing, Contact and Other Lists (SEC-56)'', originally published in the Federal Register... publicly. FOR FURTHER INFORMATION CONTACT: Barbara A. Stance, Chief Privacy Officer, Office of Information...: ``Mailing, Contact and Other Lists (SEC-56)''. As described in the original notice, the system contains...

14 CFR Sec. 1-3 - General description of system of accounts and reports.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 14 Aeronautics and Space 4 2014-01-01 2014-01-01 false General description of system of accounts and reports. Sec. 1-3 Section 1-3 Aeronautics and Space OFFICE OF THE SECRETARY, DEPARTMENT OF... reports. (a) This system of accounts and reports is designed to permit limited contraction or expansion to...

14 CFR Sec. 1-3 - General description of system of accounts and reports.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 14 Aeronautics and Space 4 2013-01-01 2013-01-01 false General description of system of accounts and reports. Sec. 1-3 Section 1-3 Aeronautics and Space OFFICE OF THE SECRETARY, DEPARTMENT OF... reports. (a) This system of accounts and reports is designed to permit limited contraction or expansion to...

14 CFR 1-3 - General description of system of accounts and reports.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 14 Aeronautics and Space 4 2012-01-01 2012-01-01 false General description of system of accounts and reports. Sec. 1-3 Section Sec. 1-3 Aeronautics and Space OFFICE OF THE SECRETARY, DEPARTMENT OF... reports. (a) This system of accounts and reports is designed to permit limited contraction or expansion to...

14 CFR Sec. 1-3 - General description of system of accounts and reports.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 14 Aeronautics and Space 4 2011-01-01 2011-01-01 false General description of system of accounts and reports. Sec. 1-3 Section 1-3 Aeronautics and Space OFFICE OF THE SECRETARY, DEPARTMENT OF... reports. (a) This system of accounts and reports is designed to permit limited contraction or expansion to...

Compositions and methods for making selenocysteine containing polypeptides

DOEpatents

Soll, Dieter; Aldag, Caroline; Hohn, Michael

2016-10-11

Non-naturally occurring tRNA.sup.Sec and methods of using them for recombinant expression of proteins engineered to include one or more selenocysteine residues are disclosed. The non-naturally occurring tRNA.sup.Sec can be used for recombinant manufacture of selenocysteine containing polypeptides encoded by mRNA without the requirement of an SECIS element. In some embodiments, selenocysteine containing polypeptides are manufactured by co-expressing a non-naturally occurring tRNA.sup.Sec a recombinant expression system, such as E. coli, with SerRS, EF-Tu, SelA, or PSTK and SepSecS, and an mRNA with at least one codon that recognizes the anticodon of the non-naturally occurring tRNA.sup.Sec.

New Type of Antimicrobial Protein Produced by the Plant Pathogen Clavibacter michiganensis subsp. michiganensis

PubMed Central

Liu, Zhanliang; Ma, Ping; Holtsmark, Ingrid; Skaugen, Morten; Eijsink, Vincent G. H.

2013-01-01

It has previously been shown that the tomato pathogen Clavibacter michiganensis subsp. michiganensis secretes a 14-kDa protein, C. michiganensis subsp. michiganensis AMP-I (CmmAMP-I), that inhibits growth of Clavibacter michiganensis subsp. sepedonicus, the causal agent of bacterial ring rot of potato. Using sequences obtained from tryptic fragments, we have identified the gene encoding CmmAMP-I and we have recombinantly produced the protein with an N-terminal intein tag. The gene sequence showed that CmmAMP-I contains a typical N-terminal signal peptide for Sec-dependent secretion. The recombinant protein was highly active, with 50% growth inhibition (IC50) of approximately 10 pmol, but was not toxic to potato leaves or tubers. CmmAMP-I does not resemble any known protein and thus represents a completely new type of bacteriocin. Due to its high antimicrobial activity and its very narrow inhibitory spectrum, CmmAMP-1 may be of interest in combating potato ring rot disease. PMID:23851100

76 FR 6801 - Agency Information Collection Activities; Submission for Office of Management and Budget Review...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-02-08

... (TEA) final rule). The regulations in Sec. 330.14 state that OTC drug products introduced into the U.S...) meet certain ``time and extent'' criteria outlined in Sec. 330.14(b). The regulations allow a TEA to be... system. TEAs must provide evidence described in Sec. 330.14(c) demonstrating that the condition is...

Advanced Numerical Methods for Simulating Nonlinear Multirate Lumped Parameter Models

DTIC Science & Technology

1991-05-01

defining a Waveform: typedef struct Waveform char *name; /* character string of the name of the variable */ double tn; /* time of the beginning of the...A State-Space Approach, Reprinted from Proc. Third Ann. Allerton Conf. Circuits and Systems Thoery , 659-668, in Computer-Aided Circuit Design...kg/n 3) 1025.9 kg/m3 @ 15* C. v Kinematic Viscosity of Water (m2/sec) 1.19x10-6 m2/sec @ 15* C. G Acceleration of Gravity (m/sec2) 9.80665 m/sec 2 L

Longitudinal monitoring of Gaussia and Nano luciferase activities to concurrently assess ER calcium homeostasis and ER stress in vivo.

PubMed

Wires, Emily S; Henderson, Mark J; Yan, Xiaokang; Bäck, Susanne; Trychta, Kathleen A; Lutrey, Molly H; Harvey, Brandon K

2017-01-01

The endoplasmic reticulum (ER) is essential to many cellular processes including protein processing, lipid metabolism and calcium storage. The ability to longitudinally monitor ER homeostasis in the same organism would offer insight into progressive molecular and cellular adaptations to physiologic or pathologic states, but has been challenging. We recently described the creation of a Gaussia luciferase (GLuc)-based secreted ER calcium-modulated protein (SERCaMP or GLuc-SERCaMP) to longitudinally monitor ER calcium homeostasis. Here we describe a complementary tool to measure the unfolded protein response (UPR), utilizing a UPRE-driven secreted Nano luciferase (UPRE-secNLuc) to examine the activating transcription factor-6 (ATF6) and inositol-requiring enzyme 1 (IRE1) pathways of the UPR. We observed an upregulation of endogenous ATF6- and XBP1-regulated genes following pharmacologically-induced ER stress that was consistent with responsiveness of the UPRE sensor. Both GLuc and NLuc-based reporters have favorable properties for in vivo studies, however, they are not easily used in combination due to overlapping substrate activities. We describe a method to measure the enzymatic activities of both reporters from a single sample and validated the approach using culture medium and rat blood samples to measure GLuc-SERCaMP and UPRE-secNLuc. Measuring GLuc and NLuc activities from the same sample allows for the robust and quantitative measurement of two cellular events or cell populations from a single biological sample. This study is the first to describe the in vivo measurement of UPRE activation by sampling blood, using an approach that allows concurrent interrogation of two components of ER homeostasis.

Vasopressin regularizes the phasic firing pattern of rat hypothalamic magnocellular vasopressin neurons.

PubMed

Gouzènes, L; Desarménien, M G; Hussy, N; Richard, P; Moos, F C

1998-03-01

Vasopressin (AVP) magnocellular neurons of hypothalamic nuclei express specific phasic firing (successive periods of activity and silence), which conditions the mode of neurohypophyseal vasopression release. In situations favoring plasmatic secretion of AVP, the hormone is also released at the somatodendritic level, at which it is believed to modulate the activity of AVP neurons. We investigated the nature of this autocontrol by testing the effects of juxtamembrane applications of AVP on the extracellular activity of presumed AVP neurons in paraventricular and supraoptic nuclei of anesthetized rats. AVP had three effects depending on the initial firing pattern: (1) excitation of faintly active neurons (periods of activity of <10 sec), which acquired or reinforced their phasic pattern; (2) inhibition of quasi-continuously active neurons (periods of silences of <10 sec), which became clearly phasic; and (3) no effect on neurons already showing an intermediate phasic pattern (active and silent periods of 10-30 sec). Consequently, AVP application resulted in a narrower range of activity patterns of the population of AVP neurons, with a Gaussian distribution centered around a mode of 57% of time in activity, indicating a homogenization of the firing pattern. The resulting phasic pattern had characteristics close to those established previously for optimal release of AVP from neurohypophyseal endings. These results suggest a new role for AVP as an optimizing factor that would foster the population of AVP neurons to discharge with a phasic pattern known to be most efficient for hormone release.

76 FR 11675 - IFR Altitudes; Miscellaneous Amendments

Federal Register 2010, 2011, 2012, 2013, 2014

2011-03-03

... action is needed because of changes occurring in the National Airspace System. These changes are designed... matters of flight safety and operational efficiency in the National Airspace System, are related to... RIVER, AK NDB BARTER ISLAND, AK NDB... 2000 From To MEA MAA Sec. 95.3000 Low Altitude RNAV Routes Sec...

14 CFR Sec. 1-4 - System of accounts coding.

Code of Federal Regulations, 2010 CFR

2010-01-01

... General Accounting Provisions Sec. 1-4 System of accounts coding. (a) A four digit control number is assigned for each balance sheet and profit and loss account. Each balance sheet account is numbered sequentially, within blocks, designating basic balance sheet classifications. The first two digits of the four...

Participation determinants in the DRG payment system of obstetrics and gynecology clinics in South Korea.

PubMed

Song, Jung-Kook; Kim, Chang-yup

2010-03-01

The Diagnosis Related Group (DRG) payment system, which has been implemented in Korea since 1997, is based on voluntary participation. Hence, the positive impact of this system depends on the participation of physicians. This study examined the factors determining participation of Korean obstetrics & gynecology (OBGYN) clinics in the DRG-based payment system. The demographic information, practice-related variables of OBGYN clinics and participation information in the DRG-based payment system were acquired from the nationwide data from 2002 to 2007 produced by the National Health Insurance Corporation and the Health Insurance Review & Assessment Service. The subjects were 336 OBGYN clinics consisting of 43 DRG clinics that had maintained their participation in 2003-2007 and 293 no-DRG (fee-for-service) clinics that had never been a DRG clinic during the same period. Logistic regression analysis was carried out to determine the factors associated with the participation of OBGYN clinics in the DRG-based payment system. The factors affecting participation of OBGYN clinics in the DRG-based payment system were as follows (p<0.05): (1) a larger number of caesarian section (c/sec) claims, (2) higher cost of a c/sec, (3) less variation in the price of a c/sec, (4) fewer days of admission for a c/sec, and (5) younger pregnant women undergoing a c/sec. These results suggest that OBGYN clinics with an economic practice pattern under a fee-for-service system are more likely to participate in the DRG-based payment system. Therefore, to ensure adequate participation of physicians, a payment system with a stronger financial incentive might be more suitable in Korea.

Electric propulsion options for the SP-100 reference mission

NASA Technical Reports Server (NTRS)

Hardy, T. L.; Rawlin, V. K.; Patterson, M. J.

1987-01-01

Analyses were performed to characterize and compare electric propulsion systems for use on a space flight demonstration of the SP-100 nuclear power system. The component masses of resistojet, arcjet, and ion thruster systems were calculated using consistent assumptions and the maximum total impulse, velocity increment, and thrusting time were determined, subject to the constraint of the lift capability of a single Space Shuttle launch. From the study it was found that for most systems the propulsion system dry mass was less than 20 percent of the available mass for the propulsion system. The maximum velocity increment was found to be up to 2890 m/sec for resistojet, 3760 m/sec for arcjet, and 23 000 m/sec for ion thruster systems. The maximum thruster time was found to be 19, 47, and 853 days for resistojet, arcjet, and ion thruster systems, respectively.

The Solar Neighborhood. XXV. Discovery of New Proper Motion Stars with 0.40 sec/yr > mu > or = 0.18 sec/yr Between Declinations -47 deg and 00 deg

NASA Technical Reports Server (NTRS)

Boyd, Mark R.; Winters, Jennifer G.; Henry, Todd J.; Jao, Wei-Chun; Finch, Charlie T.; Subasavage, John P.; Hambly, Nigel C.

2011-01-01

We present 2817 new southern proper motion systems with 0.40 sec/yr > mu > or = 0.18 sec/yr and declination between 47 deg and 00 deg. This is a continuation of the SuperCOSMOS-RECONS (SCR) proper motion searches of the southern sky. We use the same photometric relations as previous searches to provide distance estimates based on the assumption that the objects are single main-sequence stars. We find 79 new red dwarf systems predicted to be within 25 pc, including a few new components of previously known systems. Two systems--SCR 1731-2452 at 9.5 pc and SCR 1746-3214 at 9.9 pc--are anticipated to be within 10 pc. We also find 23 new white dwarf (WD) candidates with distance estimates of 15-66 pc, as well as 360 new red subdwarf candidates. With this search, we complete the SCR sweep of the southern sky for stars with mu > or = 0.18 sec/yr and R(sub 59F) < or = 16.5, resulting in a total of 5042 objects in 4724 previously unreported proper motion systems. Here we provide selected comprehensive lists from our SCR proper motion search to date, including 152 red dwarf systems estimated to be within 25 pc (9 within 10 pc), 46 WDs (10 within 25 pc), and 598 subdwarf candidates. The results of this search suggest that there are more nearby systems to be found at fainter magnitudes and lower proper motion limits than those probed so far.

High-Performance Bipropellant Engine

NASA Technical Reports Server (NTRS)

Biaglow, James A.; Schneider, Steven J.

1999-01-01

TRW, under contract to the NASA Lewis Research Center, has successfully completed over 10 000 sec of testing of a rhenium thrust chamber manufactured via a new-generation powder metallurgy. High performance was achieved for two different propellants, N2O4- N2H4 and N2O4 -MMH. TRW conducted 44 tests with N2O4-N2H4, accumulating 5230 sec of operating time with maximum burn times of 600 sec and a specific impulse Isp of 333 sec. Seventeen tests were conducted with N2O4-MMH for an additional 4789 sec and a maximum Isp of 324 sec, with a maximum firing duration of 700 sec. Together, the 61 tests totalled 10 019 sec of operating time, with the chamber remaining in excellent condition. Of these tests, 11 lasted 600 to 700 sec. The performance of radiation-cooled rocket engines is limited by their operating temperature. For the past two to three decades, the majority of radiation-cooled rockets were composed of a high-temperature niobium alloy (C103) with a disilicide oxide coating (R512) for oxidation resistance. The R512 coating practically limits the operating temperature to 1370 C. For the Earth-storable bipropellants commonly used in satellite and spacecraft propulsion systems, a significant amount of fuel film cooling is needed. The large film-cooling requirement extracts a large penalty in performance from incomplete mixing and combustion. A material system with a higher temperature capability has been matured to the point where engines are being readied for flight, particularly the 100-lb-thrust class engine. This system has powder rhenium (Re) as a substrate material with an iridium (Ir) oxidation-resistant coating. Again, the operating temperature is limited by the coating; however, Ir is capable of long-life operation at 2200 C. For Earth-storable bipropellants, this allows for the virtual elimination of fuel film cooling (some film cooling is used for thermal control of the head end). This has resulted in significant increases in specific impulse performance (15 to 20 sec). To determine the merits of a powder rhenium thrust chamber, Lewis On-Board Propulsion Branch directed TRW (under the Space Storable Rocket Technology Program and the High Pressure Earth Storable Rocket Technology Program) to design, fabricate, and test an engineering model to serve as a technology demonstrator.

Incidence of enterotoxigenic staphylococci and their toxins in foods.

PubMed

Soriano, J M; Font, G; Rico, H; Moltó, J C; Mañes, J

2002-05-01

Of 504 food samples collected from cafeterias, 19 (3.8%) yielded strains of enterotoxigenic staphylococci, and 10 (52.6%), 4 (21.1%), 3 (15.8%), and 2 (10.5%) of these strains produced enterotoxins C (SEC), D (SED), B (SEB), and A (SEA), respectively. Moreover, SEA, SEB, and SEC were isolated from three hamburger samples. Of 181 food samples collected from four restaurants before the implementation of the hazard analysis and critical control point (HACCP) system, 7 (3.9%) were found to contain enterotoxigenic strains, and SED, SEC, and SEA were produced by 4 (57.1%), 2 (28.6%), and 1 (14.3%) of these strains, respectively. One meatball sample with SEC was detected in a restaurant. After the implementation of the HACCP system in four restaurants, neither enterotoxigenic staphylococci nor enterotoxins were detected in 196 studied samples.

Flight Control System Analysis and Design for a Remotely Piloted Vehicle with Thrust Vectoring Unit.

DTIC Science & Technology

1980-12-01

about the X-axis (slug-ft 2) Ixz Product of inertia (slug-ft 2 ) ly Moi,;ent of inertia about Y-axis (slug-ft 2) Iz Moment of inertia about Z-axis (slug...domain n Load factor (g’s) P Roll rate (rad/sec) xi p Perturbation roll rate (rad/sec) Q Pitch rate (rad/sec) q Perturbation pitch rate (rad/sec...was decided to employ a scale factor of 1.75 in increasing the vertical tail area. This choice was somewhat aruitrary since no documentation could be

Understanding the Current State of Infection Prevention to Prevent Clostridium difficile Infection: A Human Factors and Systems Engineering Approach

PubMed Central

Yanke, Eric; Zellmer, Caroline; Van Hoof, Sarah; Moriarty, Helene; Carayon, Pascale; Safdar, Nasia

2015-01-01

Background Achieving and sustaining high levels of healthcare worker (HCW) compliance with contact isolation precautions is challenging. The aim of this study was to determine HCW work system barriers to, and facilitators of, adherence to contact isolation for patients with suspected or confirmed Clostridium difficile infection (CDI) using a human factors and systems engineering approach. Methods Prospective cohort study from September 2013 to November 2013 at a large academic medical center (hospital A) and an affiliated Veterans Administration (VA) hospital (hospital B). A human factors engineering (HFE) model for patient safety – the Systems Engineering Initiative for Patient Safety (SEIPS) model – was used to guide work system analysis and direct observation data collection. 288 observations were conducted. HCWs and visitors were assessed for compliance with all components of contact isolation precautions (hand hygiene, gowning, and gloving) before and after patient contact. Time required to complete contact isolation precautions was measured and adequacy of contact isolation supplies was assessed. Results Full compliance with contact isolation precautions was low at both hospitals: hospital A, 7%; hospital B, 22%. Lack of appropriate hand hygiene prior to room entry (Compliance: hospital A, 18%; hospital B, 29%) was the most common reason for lack of full compliance. More time was required for full compliance as compared to compliance with no components of contact isolation precautions before patient room entry, inside patient room, and after patient room exit (59.9 sec vs. 3.2 sec; P < .001; 507.3 sec vs. 149.7 sec; P = .006; 15.2 sec vs. 1.3 sec; P < .001). Compliance was lower when contact isolation supplies were inadequate (4% vs. 16%; P = .005). Conclusions Adherence to contact isolation precautions for CDI is a complex, time-consuming process. HFE analysis indicates multiple work system components serve as barriers and facilitators to full compliance with contact isolation precautions and should be addressed further to prevent CDI. PMID:25728149

MO-FG-BRA-02: A Feasibility Study of Integrating Breathing Audio Signal with Surface Surrogates for Respiratory Motion Management

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

Lei, Y; Zhu, X; Zheng, D

Purpose: Tracking the surrogate placed on patient skin surface sometimes leads to problematic signals for certain patients, such as shallow breathers. This in turn impairs the 4D CT image quality and dosimetric accuracy. In this pilot study, we explored the feasibility of monitoring human breathing motion by integrating breathing sound signal with surface surrogates. Methods: The breathing sound signals were acquired though a microphone attached adjacently to volunteer’s nostrils, and breathing curve were analyzed using a low pass filter. Simultaneously, the Real-time Position Management™ (RPM) system from Varian were employed on a volunteer to monitor respiratory motion including both shallowmore » and deep breath modes. The similar experiment was performed by using Calypso system, and three beacons taped on volunteer abdominal region to capture breath motion. The period of each breathing curves were calculated with autocorrelation functions. The coherence and consistency between breathing signals using different acquisition methods were examined. Results: Clear breathing patterns were revealed by the sound signal which was coherent with the signal obtained from both the RPM system and Calypso system. For shallow breathing, the periods of breathing cycle were 3.00±0.19 sec (sound) and 3.00±0.21 sec (RPM); For deep breathing, the periods were 3.49± 0.11 sec (sound) and 3.49±0.12 sec (RPM). Compared with 4.54±0.66 sec period recorded by the calypso system, the sound measured 4.64±0.54 sec. The additional signal from sound could be supplement to the surface monitoring, and provide new parameters to model the hysteresis lung motion. Conclusion: Our preliminary study shows that the breathing sound signal can provide a comparable way as the RPM system to evaluate the respiratory motion. It’s instantaneous and robust characteristics facilitate it possibly to be a either independently or as auxiliary methods to manage respiratory motion in radiotherapy.« less

Interplay of biopharmaceutics, biopharmaceutics drug disposition and salivary excretion classification systems

PubMed Central

Idkaidek, Nasir M.

2013-01-01

The aim of this commentary is to investigate the interplay of Biopharmaceutics Classification System (BCS), Biopharmaceutics Drug Disposition Classification System (BDDCS) and Salivary Excretion Classification System (SECS). BCS first classified drugs based on permeability and solubility for the purpose of predicting oral drug absorption. Then BDDCS linked permeability with hepatic metabolism and classified drugs based on metabolism and solubility for the purpose of predicting oral drug disposition. On the other hand, SECS classified drugs based on permeability and protein binding for the purpose of predicting the salivary excretion of drugs. The role of metabolism, rather than permeability, on salivary excretion is investigated and the results are not in agreement with BDDCS. Conclusion The proposed Salivary Excretion Classification System (SECS) can be used as a guide for drug salivary excretion based on permeability (not metabolism) and protein binding. PMID:24493977

Sequential Events Control System (SECS) Overview

NASA Technical Reports Server (NTRS)

Interbartolo, Michael

2009-01-01

This slide presentation will cover the Sequential Events Control System (SECS), which is the Apollo spacecraft subsystem that controls the automatically sequenced functions during the mission and during any a borts that could be performed. Included in this presentation are its general architecture, its integration into and use of the spacecraft' s other systems, and details on the functions it is responsible for c ontrolling during the mission. The objectives are to describe the system's architecture, the major components in the system, and the major system functions.

An Experimental Study of Ignition Effects and Flame Growth Over a Thin Solid Fuel in Low-Speed Concurrent Flow Using Drop-Tower Facilities

NASA Technical Reports Server (NTRS)

Pettegrew, Richard Dale

1996-01-01

An experimental study of ignition and flame growth over a thin solid fuel in oxidizer flow speeds from 0 to 10 cm/sec concurrent flow was performed. This study examined the differences between ignition using a resistively heated wire (woven in a sawtooth pattern over the leading edge of the fuel), and a straight resistively heated wire augmented by a chemical ignitor doped onto the leading edge of the fuel. Results showed that the chemical system yielded non-uniform ignition bursts, while the system using only the hotwire gave more uniform ignition. At speeds up to 2.5 cm/sec, the chemical system yielded non-uniform pyrolysis fronts, while the hotwire system gave more uniform pyrolysis fronts. At speeds of 5 cm/sec or greater, both systems gave uniform pyrolysis fronts. The chemically-ignited flames tended to become too dim to see faster than the hotwire-ignited flames, and the flame lengths were observed to be shorter (after the initial burst subsided) for the chemical system for all speeds. Flame and pyrolysis element velocities were measured. Temperature profiles for selected tests were measured using thermocouples at the fuel surface and in the gas phase. Comparisons between the flame element velocities and peak temperatures recorded in these tests with calculated spread rates and peak temperatures from a steady-state model are presented. Agreement was found to be within 20% for most flame elements for nominal velocities of 5 cm/sec and 7.5 cm/sec.

Effects of Different Lifting Cadences on Ground Reaction Forces during the Squat Exercise

NASA Technical Reports Server (NTRS)

Bentley, Jason R.; Amonette, William E.; Hagan, R. Donald

2008-01-01

The purpose of this investigation was to determine the effect of different cadences on the ground reaction force (GRF(sub R)) during the squat exercise. It is known that squats performed with greater acceleration will produce greater inertial forces; however, it is not well understood how different squat cadences affect GRF(sub R). It was hypothesized that faster squat cadences will result in greater peak GRF(sub R). METHODS: Six male subjects (30.8+/-4.4 y, 179.5+/-8.9 cm, 88.8+/-13.3 kg) with previous squat experience performed three sets of three squats using three different cadences (FC = 1 sec descent/1 sec ascent; MC = 3 sec descent/1 sec ascent; SC = 4 sec descent/2 sec ascent) with barbell mass equal to body mass. Ground reaction force was used to calculate inertial force trajectories of the body plus barbell (FI(sub system)). Forces were normalized to body mass. RESULTS: Peak GRF(sub R) and peak FI(sub system) were significantly higher in FC squats compared to MC (p=0.0002) and SC (p=0.0002). Range of GRF(sub R) and FI(sub system) were also significantly higher in FC compared to MC (p<0.05), and MC were significantly higher than SC (p<0.05). DISCUSSION: Faster squat cadences result in significantly greater peak GRF(sub R) due to the inertia of the system. GRF(sub R) was more dependent upon decent cadence than on ascent cadence. PRACTICAL APPLICATION: This study demonstrates that faster squat cadences produce greater ground reaction forces. Therefore, the use of faster squat cadences might enhance strength and power adaptations to long-term resistance exercise training. Key Words: velocity, weight training, resistive exercise

Use of the usp45 lactococcal secretion signal sequence to drive the secretion and functional expression of enterococcal bacteriocins in Lactococcus lactis.

PubMed

Borrero, Juan; Jiménez, Juan J; Gútiez, Loreto; Herranz, Carmen; Cintas, Luis M; Hernández, Pablo E

2011-01-01

Replacement of the signal peptide (SP) of the bacteriocins enterocin P (EntP) and hiracin JM79 (HirJM79), produced by Enterococcus faecium P13 and Enterococcus hirae DCH5, respectively, by the signal peptide of Usp45 (SP(usp45)), the major Sec-dependent protein secreted by Lactococcus lactis, permits the production, secretion, and functional expression of EntP and HirJM79 by L. lactis. Chimeric genes encoding the SP(usp45) fused to either mature EntP (entP), with or without the immunity gene (entiP) or to mature HirJM79 (hirJM79), with or without the immunity gene (hiriJM79), were cloned into the expression vector pMG36c, carrying the P(32) constitutive promoter, and into pNZ8048 under control of the inducible PnisA promoter. The production of EntP and HirJM79 by most of the L. lactis recombinant strains was 1.5- to 3.7-fold higher and up to 3.6-fold higher than by the E. faecium P13 and E. hirae DCH5 control strains, respectively. However, the specific antimicrobial activity of the recombinant EntP was 1.1- to 6.2-fold higher than that produced by E. faecium P13, while that of the HirJM79 was a 40% to an 89% of that produced by E. hirae DCH5. Chimeras of SP(usp45) fused to mature EntP or HirJM79 drive the production and secretion of these bacteriocins in L. lactis in the absence of specific immunity and secretion proteins. The supernatants of the recombinant L. lactis NZ9000 strains, producers of EntP, showed a much higher antimicrobial activity against Listeria spp. than that of the recombinant L. lactis NZ9000 derivatives, producers of HirJM79.

77 FR 25865 - Amendment to the International Traffic in Arms Regulations: Exemption for Temporary Export of...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-05-02

... the Automated Export System per Sec. 123.22 of this subchapter, and the articles are presented to the... System (AES) per Sec. 123.22 of this subchapter, and the articles are presented to the CBP officer for... comments and recommendations follows: Three commenting parties requested the elimination of the requirement...

Bacterial Translocation Ratchets: Shared Physical Principles with Different Molecular Implementations: How bacterial secretion systems bias Brownian motion for efficient translocation of macromolecules.

PubMed

Hepp, Christof; Maier, Berenike

2017-10-01

Secretion systems enable bacteria to import and secrete large macromolecules including DNA and proteins. While most components of these systems have been identified, the molecular mechanisms of macromolecular transport remain poorly understood. Recent findings suggest that various bacterial secretion systems make use of the translocation ratchet mechanism for transporting polymers across the cell envelope. Translocation ratchets are powered by chemical potential differences generated by concentration gradients of ions or molecules that are specific to the respective secretion systems. Bacteria employ these potential differences for biasing Brownian motion of the macromolecules within the conduits of the secretion systems. Candidates for this mechanism include DNA import by the type II secretion/type IV pilus system, DNA export by the type IV secretion system, and protein export by the type I secretion system. Here, we propose that these three secretion systems employ different molecular implementations of the translocation ratchet mechanism. © 2017 The Authors. BioEssays Published by WILEY Periodicals, Inc.

Hydraulic characteristics of an underdrained irrigation circle, Muskegon County wastewater disposal system, Michigan

USGS Publications Warehouse

McDonald, M.G.

1980-01-01

Muskegon County, Michigan, disposes of wastewater by spray irrigating farmland on its waste-disposal site. Buried drains in the highly permeable unconfined aquifer at the site control the level of the water table. Hydraulic conductivity of the aquifer and drain-leakance, the reciprocal of resistance to flow into the drains, was determined at a representative irrigation circle while calibrating a model of the groundwater flow system. Hydraulic conductivity is 0.00055 m/sec, in the north zone of the circle, and 0.00039 m/sec in the south zone. Drain leakance -6 -6 is low in both zones: 2.9 x 10m/sec in the north and 9.5 x 10 m/sec in the south. Low drain leakance is responsible for waterlogging when irrigation rates are maintained at design levels. The capacity of the study circle to accept wastewater is 35 percent less than design capacity.

Model investigation of inlet plenum flow straightening techniques for altitude test facility

NASA Technical Reports Server (NTRS)

Riddlebaugh, S. M.; Linke, H. G.

1976-01-01

An investigation was conducted to evaluate and improve the quality of the airflow to be supplied to the engine in altitude test chambers 3 and 4 of the Propulsion Systems Laboratory at the Lewis Research Center. One-twentieth-scale models of the inlet plenum chamber of the two test chambers were used in the investigation to minimize time and cost. It was possible to reduce the velocity spread in the inlet plenum from approximately 100 m/sec (330 ft/sec) to approximately 10 m/sec (30 ft/sec) through the combined use of flow diverters, multiple spaced screens, flow straighteners, and turning vanes.

Airstart performance of a digital electronic engine control system in an F-15 airplane

NASA Technical Reports Server (NTRS)

Licata, S. J.; Burcham, F. W., Jr.

1983-01-01

The airstart performance of the F100 engine equipped with a digital electronic engine control (DEEC) system was evaluated in an F-15 airplane. The DEEC system incorporates closed-loop airstart logic for improved capability. Spooldown and jet fuel starter-assisted airstarts were made over a range of airspeeds and altitudes. All jet fuel starter-assisted airstarts were successful, with airstart time varying from 35 to 60 sec. All spooldown airstarts at airspeeds of 200 knots and higher were successful; airstart times ranged from 45 sec at 250 knots to 135 sec at 200 knots. The effects of altitude on airstart success and time were small. The flight results agreed closely with previous altitude facility test results. The DEEC system provided successful airstarts at airspeeds at least 50 knots lower than the standard F100 engine control system.

The Plasmodium selenoproteome

PubMed Central

Lobanov, Alexey V.; Delgado, Cesar; Rahlfs, Stefan; Novoselov, Sergey V.; Kryukov, Gregory V.; Gromer, Stephan; Hatfield, Dolph L.; Becker, Katja; Gladyshev, Vadim N.

2006-01-01

The use of selenocysteine (Sec) as the 21st amino acid in the genetic code has been described in all three major domains of life. However, within eukaryotes, selenoproteins are only known in animals and algae. In this study, we characterized selenoproteomes and Sec insertion systems in protozoan Apicomplexa parasites. We found that among these organisms, Plasmodium and Toxoplasma utilized Sec, whereas Cryptosporidium did not. However, Plasmodium had no homologs of known selenoproteins. By searching computationally for evolutionarily conserved selenocysteine insertion sequence (SECIS) elements, which are RNA structures involved in Sec insertion, we identified four unique Plasmodium falciparum selenoprotein genes. These selenoproteins were incorrectly annotated in PlasmoDB, were conserved in other Plasmodia and had no detectable homologs in other species. We provide evidence that two Plasmodium SECIS elements supported Sec insertion into parasite and endogenous selenoproteins when they were expressed in mammalian cells, demonstrating that the Plasmodium SECIS elements are functional and indicating conservation of Sec insertion between Apicomplexa and animals. Dependence of the plasmodial parasites on selenium suggests possible strategies for antimalarial drug development. PMID:16428245

Particle velocity measurements with macroscopic fluorescence imaging in lymph tissue mimicking microfluidic phantoms

NASA Astrophysics Data System (ADS)

Hennessy, Ricky; Koo, Chiwan; Ton, Phuc; Han, Arum; Righetti, Raffaella; Maitland, Kristen C.

2011-03-01

Ultrasound poroelastography can quantify structural and mechanical properties of tissues such as stiffness, compressibility, and fluid flow rate. This novel ultrasound technique is being explored to detect tissue changes associated with lymphatic disease. We have constructed a macroscopic fluorescence imaging system to validate ultrasonic fluid flow measurements and to provide high resolution imaging of microfluidic phantoms. The optical imaging system is composed of a white light source, excitation and emission filters, and a camera with a zoom lens. The field of view can be adjusted from 100 mm x 75 mm to 10 mm x 7.5 mm. The microfluidic device is made of polydimethylsiloxane (PDMS) and has 9 channels, each 40 μm deep with widths ranging from 30 μm to 200 μm. A syringe pump was used to propel water containing 15 μm diameter fluorescent microspheres through the microchannels, with flow rates ranging from 0.5 μl/min to 10 μl/min. Video was captured at a rate of 25 frames/sec. The velocity of the microspheres in the microchannels was calculated using an algorithm that tracked the movement of the fluorescent microspheres. The imaging system was able to measure particle velocities ranging from 0.2 mm/sec to 10 mm/sec. The range of flow velocities of interest in lymph vessels is between 1 mm/sec to 10 mm/sec; therefore our imaging system is sufficient to measure particle velocity in phantoms modeling lymphatic flow.

Sec66-Dependent Regulation of Yeast Spindle-Pole Body Duplication Through Pom152

PubMed Central

Katta, Santharam S.; Chen, Jingjing; Gardner, Jennifer M.; Friederichs, Jennifer M.; Smith, Sarah E.; Gogol, Madelaine; Unruh, Jay R.; Slaughter, Brian D.; Jaspersen, Sue L.

2015-01-01

In closed mitotic systems such as Saccharomyces cerevisiae, the nuclear envelope (NE) does not break down during mitosis, so microtubule-organizing centers such as the spindle-pole body (SPB) must be inserted into the NE to facilitate bipolar spindle formation and chromosome segregation. The mechanism of SPB insertion has been linked to NE insertion of nuclear pore complexes (NPCs) through a series of genetic and physical interactions between NPCs and SPB components. To identify new genes involved in SPB duplication and NE insertion, we carried out genome-wide screens for suppressors of deletion alleles of SPB components, including Mps3 and Mps2. In addition to the nucleoporins POM152 and POM34, we found that elimination of SEC66/SEC71/KAR7 suppressed lethality of cells lacking MPS2 or MPS3. Sec66 is a nonessential subunit of the Sec63 complex that functions together with the Sec61 complex in import of proteins into the endoplasmic reticulum (ER). Cells lacking Sec66 have reduced levels of Pom152 protein but not Pom34 or Ndc1, a shared component of the NPC and SPB. The fact that Sec66 but not other subunits of the ER translocon bypass deletion mutants in SPB genes suggests a specific role for Sec66 in the control of Pom152 levels. Based on the observation that sec66∆ does not affect the distribution of Ndc1 on the NE or Ndc1 binding to the SPB, we propose that Sec66-mediated regulation of Pom152 plays an NPC-independent role in the control of SPB duplication. PMID:26510791

CoSEC: Connecting Living With a Star Research

NASA Astrophysics Data System (ADS)

Hurlburt, N.; Freeland, S.; Bose, P.; Zimdars, A.; Slater, G.

2006-12-01

The Collaborative Sun-Earth Connector (CoSEC) provide the means for heliophysics researchers to compose the data sources and processing services published by their peers into processing workflows that reliably generate publication-worthy data. It includes: composition of computational and data services into easy-to- read workflows with data quality and version traceability; straightforward translation of existing services into workflow components, and advertisement of those components to other members of the CoSEC community; annotation of published services with functional attributes to enable discovery of capabilities required by particular workflows and identify peer subgroups in the CoSEC community; and annotation of published services with nonfunctional attributes to enable selection on the basis of quality of service (QoS). We present an overview and demonstration of the CoSEC system, discuss applications, the lessons learned and future developments.

Does thrombin stimulation of human platelets proceed via a simultaneous Na/sup +/-H/sup +/ exchange

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

Davies, T.A.; Katona, E.; Vasilescu, V.

1986-03-05

Thrombin stimulation of human platelets initiates a membrane depolarization attributable to a Na/sup +/ influx into, and an alkalinization of, the cytoplasm, both of which follow a similar rapid time scale and thrombin dose dependence. These responses precede secretion of the contents of dense granules (serotonin) and, after 1 min, of lysosomes (..beta..-glucuronidase). These markers have been used to determine whether the Na/sup +/ influx and H/sup +/ efflux are sequential or simultaneous. They have examined these parameters in D/sub 2/O-Hepes buffers. NMR evidence indicates that equilibration is rapid, and virtually complete within the 3 minute pre-stimulation platelets equilibration period.more » The rate of depolarization is 70-80% slower in D/sub 2/O than in H/sub 2/O. The time to reach maximal depolarization is 5-10 sec longer, the extent of depolarization 60% inhibited, and the (H/sup +/) change 85-100% inhibited. The serotonin secretion is unaltered, and the ..beta..-glucuronidase secretion is 130-180% enhanced. 10/sup -4/ M amiloride inhibits Na/sup +/ influx, i.e. depolarization, and the pH change completely. Adjustment to pH/sub i/ 7.3 with NH/sub 4/Cl led to a 30-80% enhanced ..beta..-glucuronidase release upon thrombin exposure. These results suggest that the Na/sup +/ and H/sup +/ fluxes across the platelet membrane occur sequentially, the Na/sup +/ occurring first. Furthermore, granule secretion, previously shown by us to be independent of the existent Na/sup +/ gradient, depends on the cytoplasmic K/sup +/ and H/sup +/ concentrations.« less

Pressure Loss Predictions of the Reactor Simulator Subsystem at NASA GRC

NASA Technical Reports Server (NTRS)

Reid, Terry V.

2015-01-01

Testing of the Fission Power System (FPS) Technology Demonstration Unit (TDU) is being conducted at NASA GRC. The TDU consists of three subsystems: the Reactor Simulator (RxSim), the Stirling Power Conversion Unit (PCU), and the Heat Exchanger Manifold (HXM). An Annular Linear Induction Pump (ALIP) is used to drive the working fluid. A preliminary version of the TDU system (which excludes the PCU for now), is referred to as the RxSim subsystem and was used to conduct flow tests in Vacuum Facility 6 (VF 6). In parallel, a computational model of the RxSim subsystem was created based on the CAD model and was used to predict loop pressure losses over a range of mass flows. This was done to assess the ability of the pump to meet the design intent mass flow demand. Measured data indicates that the pump can produce 2.333 kg/sec of flow, which is enough to supply the RxSim subsystem with a nominal flow of 1.75 kg/sec. Computational predictions indicated that the pump could provide 2.157 kg/sec (using the Spalart-Allmaras turbulence model), and 2.223 kg/sec (using the k-? turbulence model). The computational error of the predictions for the available mass flow is -0.176 kg/sec (with the S-A turbulence model) and -0.110 kg/sec (with the k-epsilon turbulence model) when compared to measured data.

Successful ShakeAlert Performance for the Napa Quake

NASA Astrophysics Data System (ADS)

Allen, R. M.; Given, D. D.; Heaton, T. H.; Vidale, J. E.

2014-12-01

ShakeAlert, the demonstration earthquake early warning system, developed by the USGS, UC Berkeley, Caltech, ETH, and the University of Washington, functioned as expected for the August 24, 2014, M6.0 Napa earthquake. The first ShakeAlert was generated by the ElarmS algorithm 5.1 sec after the origin time of the earthquake, and 3.3 sec after the P-wave arrived at the closest station 6.5 km from the epicenter. This initial alert, based on P-wave triggers from four stations, estimated the magnitude to be 5.7. The warning was received at the UC Berkeley Seismological Laboratory 5 seconds before the S-wave and about 10 sec prior to the onset of the strongest shaking. ShakeAlert beta-testers across the San Francisco Bay Area simultaneously received the alert, including the San Francisco 911 center with 8 sec warning, and the BART train system. BART has implemented an automated train-stopping system that was activated (although no trains were running at 3:20 am). With the available network geometry and communications, the blind zone of the first alert had a radius of 16 km. The four stations that contributed to the first alert all encapsulate data into 1-second packets, but the latency in transmitting data to the processing center ranged from 0.27 to 2.62 seconds. If all the stations were to deliver data in 0.27 seconds, then the alert would have been available 2.3 sec sooner and the blind zone would be reduced to about 8 km. This would also mean that the city of Napa would have received about 1 second of warning. The magnitude estimate and event location were accurate from the initial alert onwards. The magnitude estimate did first increase to 5.8 and then dip to 5.4 2.6 sec after the initial alert, stayed at that level for 2 sec, and then returned to 5.7. The final magnitude estimate was 6.0, consistent with the ANSS catalog.

Tracking and data system support for the Mariner Mars 1971 mission. Prelaunch phase through first trajectory correction maneuver, volume 1

NASA Technical Reports Server (NTRS)

Laeser, R. P.; Textor, G. P.; Kelly, L. B.; Kelly, M.

1972-01-01

The DSN command system provided the capability to enter commands in a computer at the deep space stations for transmission to the spacecraft. The high-rate telemetry system operated at 16,200 bits/sec. This system will permit return to DSS 14 of full-resolution television pictures from the spacecraft tape recorder, plus the other science experiment data, during the two playback periods of each Goldstone pass planned for each corresponding orbit. Other features included 4800 bits/sec modem high-speed data lines from all deep space stations to Space Flight Operations Facility (SFOF) and the Goddard Space Flight Center, as well as 50,000 bits/sec wideband data lines from DSS 14 to the SFOF, thus providing the capability for data flow of two 16,200 bits/sec high-rate telemetry data streams in real time. The TDS performed prelaunch training and testing and provided support for the Mariner Mars 1971/Mission Operations System training and testing. The facilities of the ETR, DSS 71, and stations of the MSFN provided flight support coverage at launch and during the near-earth phase. The DSSs 12, 14, 41, and 51 of the DSN provided the deep space phase support from 30 May 1971 through 4 June 1971.

Application of a Saddle-Type Eddy Current Sensor in Steel Ball Surface-Defect Inspection.

PubMed

Zhang, Huayu; Zhong, Mingming; Xie, Fengqin; Cao, Maoyong

2017-12-05

Steel ball surface-defect inspection was performed by using a new saddle-type eddy current sensor (SECS), which included a saddle coil and a signal conditioning circuit. The saddle coil was directly wound on the steel ball's outer bracket in a semi-circumferential direction. Driven by a friction wheel, the test steel ball rotated in a one-dimensional direction, such that the steel ball surface was fully scanned by the SECS. There were two purposes for using the SECS in the steel ball inspection system: one was to reduce the complexity of the unfolding wheel of the surface deployment mechanism, and the other was to reduce the difficulty of sensor processing and installation. Experiments were carried out on bearing steel balls in diameter of 8 mm with three types of representative and typical defects by using the SECS, and the results showed that the inspection system can detect surface defects as small as 0.05 mm in width and 0.1 mm in depth with high-repetition detection accuracy, and the detection efficiency of 5 pcs/s, which meet the requirement for inspecting ISO grade 10 bearing steel balls. The feasibility of detecting steel ball surface defects by SECS was verified.

Kinetic resolution of sec-alcohols catalysed by Candida antarctica lipase B displaying Pichia pastoris whole-cell biocatalyst.

PubMed

Zhang, Kun; Pan, Zhiyou; Diao, Zhoujian; Liang, Shuli; Han, Shuangyan; Zheng, Suiping; Lin, Ying

2018-03-01

Kinetic resolution of sec-alcohols is a green process with biocatalyst. Candida antarctica lipase B (CALB) displayed on Pichia pastoris cell-surface (Pp-CALB) was characterized in kinetic resolution of sec-alcohols with different structures. The reaction parameters including acyl donors, molar ratio of substrates, solvents and temperatures were examined with 2-octanol as model substrate. 47.4% molar conversion of 2-octanol and 99.7% ee p were obtained after a 5h reaction with Pp-CALB, and 90% of its original activity still remained after being reused for 10 cycles. Pp-CALB was then used to several sec-alcohols and it showed great enzymatic activity and enantioselectivity to all tested sec-alcohols, more than 93.1% of ee p . The enantioselective characteristics of Pp-CALB catalysed sec-alcohols with different structures were compared with Novozyme 435 which was almost the same. Solvent free system as one way of green chemistry was applied to Pp-CALB and Pp-CALB showed great catalytic activity and enantioselectivity. Pp-CALB was potential biocatalyst of high enzymatic activity and enantioselectivity using in resolution of sec-alcohols. Copyright © 2017. Published by Elsevier Inc.

14 CFR Sec. 1-7 - Interpretation of accounts.

Code of Federal Regulations, 2010 CFR

2010-01-01

... PROCEEDINGS) ECONOMIC REGULATIONS UNIFORM SYSTEM OF ACCOUNTS AND REPORTS FOR LARGE CERTIFICATED AIR CARRIERS General Accounting Provisions Sec. 1-7 Interpretation of accounts. To the end that uniform accounting may be maintained, questions involving matters of accounting significance which are not clearly provided...

The Design of NetSecLab: A Small Competition-Based Network Security Lab

ERIC Educational Resources Information Center

Lee, C. P.; Uluagac, A. S.; Fairbanks, K. D.; Copeland, J. A.

2011-01-01

This paper describes a competition-style of exercise to teach system and network security and to reinforce themes taught in class. The exercise, called NetSecLab, is conducted on a closed network with student-formed teams, each with their own Linux system to defend and from which to launch attacks. Students are expected to learn how to: 1) install…

Using two-dimensional correlation size exclusion chromatography (2D-CoSEC) to explore the size-dependent heterogeneity of humic substances for copper binding.

PubMed

Lee, Yun-Kyung; Hur, Jin

2017-08-01

Knowledge of the heterogeneous distribution of humic substances (HS) reactivities along a continuum of molecular weight (MW) is crucial for the systems where the HS MW is subject to change. In this study, two dimensional correlation spectroscopy combined with size exclusion chromatography (2D-CoSEC) was first utilized to obtain a continuous and heterogeneous presence of copper binding characteristics within bulk HS with respect to MW. HS solutions with varying copper concentrations were directly injected into a size exclusion chromatography (SEC) system with Tris-HCl buffer as a mobile phase. Several validation tests confirmed neither structural disruption of HS nor competition effect of the mobile phase used. Similar to batch systems, fluorescence quenching was observed in the chromatograms over a wide range of HS MW. 2D-CoSEC maps of a soil-derived HS (Elliot soil humic acid) showed the greater fluorescence quenching degrees with respect to the apparent MW on the order of 12500 Da > 10600 Da > 7000 Da > 15800 Da. The binding constants calculated based on modified Stern-Volmer equation were consistent with the 2D-CoSEC results. More heterogeneity of copper binding affinities within bulk HS was found for the soil-derived HS versus an aquatic HS. The traditional fluorescence quenching titration method using ultrafiltered HS size fractions failed to delineate detailed distribution of the copper binding characteristics, exhibiting a much shorter range of the binding constants than those obtained from the 2D-CoSEC. Our proposed technique demonstrated a great potential to describe metal binding characteristics of HS at high MW resolution, providing a clear picture of the size-dependent metal-HS interactions. Copyright © 2017 Elsevier Ltd. All rights reserved.

14 CFR Sec. 1-8 - Address for reports and correspondence.

Code of Federal Regulations, 2010 CFR

2010-01-01

... (AVIATION PROCEEDINGS) ECONOMIC REGULATIONS UNIFORM SYSTEM OF ACCOUNTS AND REPORTS FOR LARGE CERTIFICATED AIR CARRIERS General Accounting Provisions Sec. 1-8 Address for reports and correspondence. All... Information, K-25, U.S. Department of Transportation, 1200 New Jersey Avenue, SE., Washington, DC 20590...

33 CFR 401.19 - Disposal and discharge systems.

Code of Federal Regulations, 2014 CFR

2014-07-01

... discharge pipes. (d) Burning of shipboard garbage is prohibited between CIP 2 & Cardinal and between CIP 15 and CIP 16. (68 Stat. 93-96, 33 U.S.C. 981-990, as amended and secs. 4, 5, 6, 7, 8, 12 and 13 of sec...

Effects of different sonic activation protocols on debridement efficacy in teeth with single-rooted canals

PubMed Central

Niu, Li-na; Luo, Xiao-juan; Li, Guo-hua; Bortoluzzi, Eduardo A.; Mao, Jing; Chen, Ji-hua; Gutmann, James L.; Pashley, David H.; Tay, Franklin R.

2014-01-01

Objectives The effects of different EndoActivator® (EA) sonic activation protocols on root canal debridement efficacy were examined. Methods Root canals in 48 single-rooted teeth were instrumented, irrigated initially with NaOCl and divided into 6 groups (N=8) based on the application time of QMix (antimicrobial calcium-chelating irrigant), and the time and sequence of EA irrigant activation - Positive Control: 90 sec QMix; Negative Control: 90 sec saline; Group 1A: 15 sec QMix + 15 sec QMix with EA-activation; Group 1B: 30 sec QMix + 30 sec of QMix with EA-activation; Group 2A: 15 sec QMix with EA-activation + 15 sec QMix; Group 2B: 30 sec QMix with EA-activation + 30 sec QMix. Split roots were examined with scanning electron microscopy for assignment of smear and debris scores in locations along the coronal, middle and apical thirds of the canals. The overall cleanliness of pooled canal locations in the Positive Control and the 4 experimental groups were compared with chi-square tests. Results Significant differences were detected among the 5 groups (p<0.001). Post-hoc pairwise comparisons indicated that the overall canal cleanliness was in the order (from best to worst): 1B = 2B > 2A > 1A > Positive Control. Completely clean canals could not be achieved due to the absence of continuous irrigant flow for EA to clear intraradicular debris. Conclusions Irrespective of the sonic activation sequence, irrigant activation for 30 seconds during a 60-second period of QMix application appears to maximize the smear layer and debris removal potential of the EndoActivator® system. PMID:24878251

Size exclusion chromatography and viscometry in paper degradation studies. New Mark-Houwink coefficients for cellulose in cupri-ethylenediamine.

PubMed

Łojewski, Tomasz; Zieba, Katarzyna; Lojewska, Joanna

2010-10-15

The paper deals with the application of size exclusion chromatography (SEC) for the studies of paper degradation phenomena. The goal is to solve some of the technical problems connected with the calibration of multi-detector SEC system and to find the correlation between SEC and viscometric results of degree of polymerization of cellulose. The results gathered for the paper samples degraded by acidic air pollutant (NO(2)) are used as an example of SEC-MALLS application. From the correlation between intrinsic viscosities and absolute value of molecular masses obtained with SEC/MALLS (Multi Angle Laser Light Scattering) technique, Mark-Houwink coefficients for cellulose in cupri-ethylenediamine solution were determined. Thus obtained coefficients were used for the determination of viscometric degree of polymerization (molecular mass) of the aged samples. An excellent correlation was found between the chromatographic values of molecular masses obtained with SEC-UV/VIS detection and the viscometric ones utilizing the improved values of Mark-Houwink coefficients. Copyright © 2010 Elsevier B.V. All rights reserved.

Pressure Loss Predictions of the Reactor Simulator Subsystem at NASA Glenn Research Center

NASA Technical Reports Server (NTRS)

Reid, Terry V.

2016-01-01

Testing of the Fission Power System (FPS) Technology Demonstration Unit (TDU) is being conducted at NASA Glenn Research Center. The TDU consists of three subsystems: the reactor simulator (RxSim), the Stirling Power Conversion Unit (PCU), and the heat exchanger manifold (HXM). An annular linear induction pump (ALIP) is used to drive the working fluid. A preliminary version of the TDU system (which excludes the PCU for now) is referred to as the "RxSim subsystem" and was used to conduct flow tests in Vacuum Facility 6 (VF 6). In parallel, a computational model of the RxSim subsystem was created based on the computer-aided-design (CAD) model and was used to predict loop pressure losses over a range of mass flows. This was done to assess the ability of the pump to meet the design intent mass flow demand. Measured data indicates that the pump can produce 2.333 kg/sec of flow, which is enough to supply the RxSim subsystem with a nominal flow of 1.75 kg/sec. Computational predictions indicated that the pump could provide 2.157 kg/sec (using the Spalart-Allmaras (S?A) turbulence model) and 2.223 kg/sec (using the k- turbulence model). The computational error of the predictions for the available mass flow is ?0.176 kg/sec (with the S-A turbulence model) and -0.110 kg/sec (with the k- turbulence model) when compared to measured data.

78 FR 37713 - General Regulations; National Park System, Demonstrations, Sale or Distribution of Printed Matter

Federal Register 2010, 2011, 2012, 2013, 2014

2013-06-24

... Statistical Abstract, in 2012 there were approximately 282 million visits to units of the National Park System...: 16 U.S.C. 1, 3, 9a, 462(k). 0 2. In Sec. 2.51 revise the introductory text of paragraph (f) to read...), paragraph (b)(4), and the introductory text of paragraph (e) to read as follows: Sec. 2.52 Sale or...

The Seasat SAR Wind and Ocean Wave Monitoring Capabilities: A case study for pass 1339m

NASA Technical Reports Server (NTRS)

Beal, R. C.

1980-01-01

A well organized low energy 11 sec. swell system off the East Coast of the U.S. was detected with the Seasat Synthetic Aperture Radar and successfully tracked from deep water, across the continental shelf, and into shallow water. In addition, a less organized 7 sec. system was tentatively identified in the imagery. Both systems were independently confirmed with simultaneous wave spectral measurements from a research pier, aircraft laser profilometer data, and Fleet Numerical Spectral Ocean Wave Models.

El Nino and the Global Ocean Observing System

NASA Technical Reports Server (NTRS)

Halpern, David

1999-01-01

Until a decade ago, an often-quoted expression in oceanography is that very few observations are recorded throughout the ocean. Now, the sentiment is no longer valid in the uppermost 10% of the tropical Pacific Ocean nor at the surface of the global ocean. One of the remarkable legacies of the 1985-1994 Tropical Oceans Global Atmosphere (TOGA) Program is an in situ marine meteorological and upper oceanographic measurement array throughout the equatorial Pacific to monitor the development and maintenance of El Nino episodes. The TOGA Observing System, which initially consisted of moored- and drifting-buoy arrays, a network of commercial ships, and coastal and island stations, now includes a constellation of satellites and data-assimilating models to simulate subsurface oceanographic conditions. The El Nino and La Nina tropical Pacific Ocean observing system represents the initial phase of an integrated global ocean observing system. Remarkable improvements have been made in ocean model simulation of subsurface currents, but some problems persist. For example, the simulation of the South Equatorial Current (SEC) remains an important challenge in the 2S-2N Pacific equatorial wave guide. During El Nino the SEC at the equator is reduced and sometimes the direction is reversed, becoming eastward. Both conditions allow warm water stored in the western Pacific to invade the eastern region, creating an El Nino episode. Assimilation of data is a tenet of faith to correct simulation errors caused by deficiencies in surface fluxes (especially wind stress) and parameterizations of subgrid-scale physical processes. In the first of two numerical experiments, the Pacific SEC was simulated with and without assimilation of subsurface temperature data. Along the equator, a very weak SEC occurred throughout the eastern Pacific, independent of assimilation of data. However, as displayed in the diagram, in the western Pacific there was no satisfactory agreement between the two simulations. To help determine reliability of the simulated SEC in the western Pacific, current measurements recorded during the 9-19 October 1994 voyage of the French research vessel L'Atalante are also shown in the diagram. With data assimilation, the simulated SEC was in much better agreement with L'Atalante observations. The simulated SEC with data assimilation was far from perfect, in part because of the sparsity of subsurface temperature observations. In the next experiment, TOPEX/POSEIDON sea surface height data in combination with subsurface temperatures will be assimilated to assess further improvement of the simulation of the SEC.

In vitro membrane protein synthesis inside Sec translocon-reconstituted cell-sized liposomes

PubMed Central

Ohta, Naoki; Kato, Yasuhiko; Watanabe, Hajime; Mori, Hirotada; Matsuura, Tomoaki

2016-01-01

Protein synthesis using an in vitro transcription-translation system (IVTT) inside cell-sized liposomes has become a valuable tool to study the properties of biological systems under cell-mimicking conditions. However, previous liposome systems lacked the machinery for membrane protein translocation. Here, we reconstituted the translocon consisting of SecYEG from Escherichia coli inside cell-sized liposomes. The cell-sized liposomes also carry the reconstituted IVTT, thereby providing a cell-mimicking environment for membrane protein synthesis. By using EmrE, a multidrug transporter from E. coli, as a model membrane protein, we found that both the amount and activity of EmrE synthesized inside the liposome is increased approximately three-fold by incorporating the Sec translocon. The topological change of EmrE induced by the translocon was also identified. The membrane integration of 6 out of 9 E. coli inner membrane proteins that was tested was increased by incorporation of the translocon. By introducing the Sec translocon, the membrane integration efficiency of the membrane protein of interest was increased, and enabled the integration of membrane proteins that otherwise cannot be inserted. In addition, this work represents an essential step toward the construction of an artificial cell through a bottom-up approach. PMID:27808179

Application of a Saddle-Type Eddy Current Sensor in Steel Ball Surface-Defect Inspection

PubMed Central

Zhong, Mingming; Xie, Fengqin; Cao, Maoyong

2017-01-01

Steel ball surface-defect inspection was performed by using a new saddle-type eddy current sensor (SECS), which included a saddle coil and a signal conditioning circuit. The saddle coil was directly wound on the steel ball’s outer bracket in a semi-circumferential direction. Driven by a friction wheel, the test steel ball rotated in a one-dimensional direction, such that the steel ball surface was fully scanned by the SECS. There were two purposes for using the SECS in the steel ball inspection system: one was to reduce the complexity of the unfolding wheel of the surface deployment mechanism, and the other was to reduce the difficulty of sensor processing and installation. Experiments were carried out on bearing steel balls in diameter of 8 mm with three types of representative and typical defects by using the SECS, and the results showed that the inspection system can detect surface defects as small as 0.05 mm in width and 0.1 mm in depth with high-repetition detection accuracy, and the detection efficiency of 5 pcs/s, which meet the requirement for inspecting ISO grade 10 bearing steel balls. The feasibility of detecting steel ball surface defects by SECS was verified. PMID:29206154

78 FR 76614 - Resolution of Systemically Important Financial Institutions: The Single Point of Entry Strategy

Federal Register 2010, 2011, 2012, 2013, 2014

2013-12-18

... banking agency or other primary financial regulatory agency; (5) retention of accounting and valuation... financial company's independent accounting firm. Additional controls may be imposed by the FDIC as... principles and applicable SEC requirements. The FDIC has consulted with the SEC regarding the accounting...

14 CFR Sec. 2-2 - Basis of allocation between entities.

Code of Federal Regulations, 2013 CFR

2013-01-01

.... 2-2 Section 2-2 Aeronautics and Space OFFICE OF THE SECRETARY, DEPARTMENT OF TRANSPORTATION... AIR CARRIERS General Accounting Provisions Sec. 2-2 Basis of allocation between entities. (a) The... prescribed by this system of accounts and reports, and (2) with respect to separate ventures, in the...

14 CFR 2-2 - Basis of allocation between entities.

Code of Federal Regulations, 2012 CFR

2012-01-01

.... 2-2 Section Sec. 2-2 Aeronautics and Space OFFICE OF THE SECRETARY, DEPARTMENT OF TRANSPORTATION... AIR CARRIERS General Accounting Provisions Sec. 2-2 Basis of allocation between entities. (a) The... prescribed by this system of accounts and reports, and (2) with respect to separate ventures, in the...

14 CFR Sec. 2-2 - Basis of allocation between entities.

Code of Federal Regulations, 2014 CFR

2014-01-01

.... 2-2 Section 2-2 Aeronautics and Space OFFICE OF THE SECRETARY, DEPARTMENT OF TRANSPORTATION... AIR CARRIERS General Accounting Provisions Sec. 2-2 Basis of allocation between entities. (a) The... prescribed by this system of accounts and reports, and (2) with respect to separate ventures, in the...

77 FR 40493 - Export Administration Regulations

Federal Register 2010, 2011, 2012, 2013, 2014

2012-07-10

... 760 Export Administration Regulations CFR Correction In Title 15 of the Code of Federal Regulations... Sec. 740.1, correctly revise the heading of paragraph (d) to read ``Shippers Export Declaration or Automated Export System Record''. 0 2. On page 321, in Sec. 742.15, move the note to introductory paragraph...

Solving Boltzmann and Fokker-Planck Equations Using Sparse Representation

DTIC Science & Technology

2011-05-31

material science. We have com- puted the electronic structure of 2D quantum dot system, and compared the efficiency with the benchmark software OCTOPUS . For...one self-consistent iteration step with 512 electrons, OCTOPUS costs 1091 sec, and selected inversion costs 9.76 sec. The algorithm exhibits

Genetic markers of insulin sensitivity and insulin secretion are associated with spontaneous postnatal growth and response to growth hormone treatment in short SGA children: the North European SGA Study (NESGAS).

PubMed

Jensen, Rikke Beck; Thankamony, Ajay; Day, Felix; Scott, Robert A; Langenberg, Claudia; Kirk, Jeremy; Donaldson, Malcolm; Ivarsson, Sten-A; Söder, Olle; Roche, Edna; Hoey, Hilary; Juul, Anders; Ong, Ken K; Dunger, David B

2015-03-01

The wide heterogeneity in the early growth and metabolism of children born small for gestational age (SGA), both before and during GH therapy, may reflect common genetic variations related to insulin secretion or sensitivity. Combined multiallele single nucleotide polymorphism scores with known associations with insulin sensitivity or insulin secretion were analyzed for their relationships with spontaneous postnatal growth and first-year responses to GH therapy in 96 short SGA children. The insulin sensitivity allele score (GS-InSens) was positively associated with spontaneous postnatal weight gain (regression coefficient [B]: 0.12 SD scores per allele; 95% confidence interval [CI], 0.01-0.23; P = .03) and also in response to GH therapy with first-year height velocity (B: 0.18 cm/y per allele; 95% CI, 0.02-0.35; P = .03) and change in IGF-1 (B: 0.17 SD scores per allele; 95% CI, 0.00-0.32; P = .03). The association with first-year height velocity was independent of reported predictors of response to GH therapy (adjusted P = .04). The insulin secretion allele score (GS-InSec) was positively associated with spontaneous postnatal height gain (B: 0.15; 95% CI, 0.01-0.30; P = .03) and disposition index both before (B: 0.02; 95% CI, 0.00-0.04; P = .04) and after 1 year of GH therapy (B: 0.03; 95% CI, 0.01-0.05; P = .002), but not with growth and IGF-1 responses to GH therapy. Neither of the allele scores was associated with size at birth. Genetic allele scores indicative of insulin sensitivity and insulin secretion were associated with spontaneous postnatal growth and responses to GH therapy in short SGA children. Further pharmacogenetic studies may support the rationale for adjuvant therapies by informing the mechanisms of treatment response.

Hepatitis C Virus Lipoviroparticles Assemble in the Endoplasmic Reticulum (ER) and Bud off from the ER to the Golgi Compartment in COPII Vesicles.

PubMed

Syed, Gulam H; Khan, Mohsin; Yang, Song; Siddiqui, Aleem

2017-08-01

Hepatitis C virus (HCV) exists as a lipoprotein-virus hybrid lipoviroparticle (LVP). In vitro studies have demonstrated the importance of apolipoproteins in HCV secretion and infectivity, leading to the notion that HCV coopts the secretion of very-low-density lipoprotein (VLDL) for its egress. However, the mechanisms involved in virus particle assembly and egress are still elusive. The biogenesis of VLDL particles occurs in the endoplasmic reticulum (ER), followed by subsequent lipidation in the ER and Golgi compartment. The secretion of mature VLDL particles occurs through the Golgi secretory pathway. HCV virions are believed to latch onto or fuse with the nascent VLDL particle in either the ER or the Golgi compartment, resulting in the generation of LVPs. In our attempt to unravel the collaboration between HCV and VLDL secretion, we studied HCV particles budding from the ER en route to the Golgi compartment in COPII vesicles. Biophysical characterization of COPII vesicles fractionated on an iodixanol gradient revealed that HCV RNA is enriched in the highly buoyant COPII vesicle fractions and cofractionates with apolipoprotein B (ApoB), ApoE, and the HCV core and envelope proteins. Electron microscopy of immunogold-labeled microsections revealed that the HCV envelope and core proteins colocalize with apolipoproteins and HCV RNA in Sec31-coated COPII vesicles. Ultrastructural analysis also revealed the presence of HCV structural proteins, RNA, and apolipoproteins in the Golgi stacks. These findings support the hypothesis that HCV LVPs assemble in the ER and are transported to the Golgi compartment in COPII vesicles to embark on the Golgi secretory route. IMPORTANCE HCV assembly and release accompany the formation of LVPs that circulate in the sera of HCV patients and are also produced in an in vitro culture system. The pathway of HCV morphogenesis and secretion has not been fully understood. This study investigates the exact site where the association of HCV virions with host lipoproteins occurs. Using immunoprecipitation of COPII vesicles and immunogold electron microscopy (EM), we characterize the existence of LVPs that cofractionate with lipoproteins, viral proteins, RNA, and vesicular components. Our results show that this assembly occurs in the ER, and LVPs thus formed are carried through the Golgi network by vesicular transport. This work provides a unique insight into the HCV LVP assembly process within infected cells and offers opportunities for designing antiviral therapeutic cellular targets. Copyright © 2017 American Society for Microbiology.

Hepatitis C Virus Lipoviroparticles Assemble in the Endoplasmic Reticulum (ER) and Bud off from the ER to the Golgi Compartment in COPII Vesicles

PubMed Central

Khan, Mohsin; Yang, Song

2017-01-01

ABSTRACT Hepatitis C virus (HCV) exists as a lipoprotein-virus hybrid lipoviroparticle (LVP). In vitro studies have demonstrated the importance of apolipoproteins in HCV secretion and infectivity, leading to the notion that HCV coopts the secretion of very-low-density lipoprotein (VLDL) for its egress. However, the mechanisms involved in virus particle assembly and egress are still elusive. The biogenesis of VLDL particles occurs in the endoplasmic reticulum (ER), followed by subsequent lipidation in the ER and Golgi compartment. The secretion of mature VLDL particles occurs through the Golgi secretory pathway. HCV virions are believed to latch onto or fuse with the nascent VLDL particle in either the ER or the Golgi compartment, resulting in the generation of LVPs. In our attempt to unravel the collaboration between HCV and VLDL secretion, we studied HCV particles budding from the ER en route to the Golgi compartment in COPII vesicles. Biophysical characterization of COPII vesicles fractionated on an iodixanol gradient revealed that HCV RNA is enriched in the highly buoyant COPII vesicle fractions and cofractionates with apolipoprotein B (ApoB), ApoE, and the HCV core and envelope proteins. Electron microscopy of immunogold-labeled microsections revealed that the HCV envelope and core proteins colocalize with apolipoproteins and HCV RNA in Sec31-coated COPII vesicles. Ultrastructural analysis also revealed the presence of HCV structural proteins, RNA, and apolipoproteins in the Golgi stacks. These findings support the hypothesis that HCV LVPs assemble in the ER and are transported to the Golgi compartment in COPII vesicles to embark on the Golgi secretory route. IMPORTANCE HCV assembly and release accompany the formation of LVPs that circulate in the sera of HCV patients and are also produced in an in vitro culture system. The pathway of HCV morphogenesis and secretion has not been fully understood. This study investigates the exact site where the association of HCV virions with host lipoproteins occurs. Using immunoprecipitation of COPII vesicles and immunogold electron microscopy (EM), we characterize the existence of LVPs that cofractionate with lipoproteins, viral proteins, RNA, and vesicular components. Our results show that this assembly occurs in the ER, and LVPs thus formed are carried through the Golgi network by vesicular transport. This work provides a unique insight into the HCV LVP assembly process within infected cells and offers opportunities for designing antiviral therapeutic cellular targets. PMID:28515296

Prolactin promotes normal liver growth, survival, and regeneration in rodents: effects on hepatic IL-6, suppressor of cytokine signaling-3, and angiogenesis.

PubMed

Moreno-Carranza, Bibiana; Goya-Arce, Maite; Vega, Claudia; Adán, Norma; Triebel, Jakob; López-Barrera, Fernando; Quintanar-Stéphano, Andrés; Binart, Nadine; Martínez de la Escalera, Gonzalo; Clapp, Carmen

2013-10-01

Prolactin (PRL) is a potent liver mitogen and proangiogenic hormone. Here, we used hyperprolactinemic rats and PRL receptor-null mice (PRLR(-/-)) to study the effect of PRL on liver growth and angiogenesis before and after partial hepatectomy (PH). Liver-to-body weight ratio (LBW), hepatocyte and sinusoidal endothelial cell (SEC) proliferation, and hepatic expression of VEGF were measured before and after PH in hyperprolactinemic rats, generated by placing two anterior pituitary glands (AP) under the kidney capsule. Also, LBW and hepatic expression of IL-6, as well as suppressor of cytokine signaling-3 (SOCS-3), were evaluated in wild-type and PRLR(-/-) mice before and after PH. Hyperprolactinemia increased the LBW, the proliferation of hepatocytes and SECs, and VEGF hepatic expression. Also, liver regeneration was increased in AP-grafted rats and was accompanied by elevated hepatocyte and SEC proliferation, and VEGF expression compared with nongrafted controls. Lowering circulating PRL levels with CB-154, an inhibitor of AP PRL secretion, prevented AP-induced stimulation of liver growth. Relative to wild-type animals, PRLR(-/-) mice had smaller livers, and soon after PH, they displayed an approximately twofold increased mortality and elevated and reduced hepatic IL-6 and SOCS-3 expression, respectively. However, liver regeneration was improved in surviving PRLR(-/-) mice. PRL stimulates normal liver growth, promotes survival, and regulates liver regeneration by mechanisms that may include hepatic downregulation of IL-6 and upregulation of SOCS-3, increased hepatocyte proliferation, and angiogenesis. PRL contributes to physiological liver growth and has potential clinical utility for ensuring survival and regulating liver mass in diseases, injuries, or surgery of the liver.

DBSecSys: a database of Burkholderia mallei secretion systems.

PubMed

Memišević, Vesna; Kumar, Kamal; Cheng, Li; Zavaljevski, Nela; DeShazer, David; Wallqvist, Anders; Reifman, Jaques

2014-07-16

Bacterial pathogenicity represents a major public health concern worldwide. Secretion systems are a key component of bacterial pathogenicity, as they provide the means for bacterial proteins to penetrate host-cell membranes and insert themselves directly into the host cells' cytosol. Burkholderia mallei is a Gram-negative bacterium that uses multiple secretion systems during its host infection life cycle. To date, the identities of secretion system proteins for B. mallei are not well known, and their pathogenic mechanisms of action and host factors are largely uncharacterized. We present the Database of Burkholderia malleiSecretion Systems (DBSecSys), a compilation of manually curated and computationally predicted bacterial secretion system proteins and their host factors. Currently, DBSecSys contains comprehensive experimentally and computationally derived information about B. mallei strain ATCC 23344. The database includes 143 B. mallei proteins associated with five secretion systems, their 1,635 human and murine interacting targets, and the corresponding 2,400 host-B. mallei interactions. The database also includes information about 10 pathogenic mechanisms of action for B. mallei secretion system proteins inferred from the available literature. Additionally, DBSecSys provides details about 42 virulence attenuation experiments for 27 B. mallei secretion system proteins. Users interact with DBSecSys through a Web interface that allows for data browsing, querying, visualizing, and downloading. DBSecSys provides a comprehensive, systematically organized resource of experimental and computational data associated with B. mallei secretion systems. It provides the unique ability to study secretion systems not only through characterization of their corresponding pathogen proteins, but also through characterization of their host-interacting partners.The database is available at https://applications.bhsai.org/dbsecsys.

Flightworthy active control landing gear for a supersonic aircraft

NASA Technical Reports Server (NTRS)

Ross, I.

1980-01-01

A flightworthy active control landing gear system for a supersonic aircraft was designed to minimize aircraft loads during takeoff, impact, rollout, and taxi. The design consists of hydromechanical modifications to the existing gear and the development of a fail-safe electronic controller. analytical RESULTS INDICATE that for an aircraft sink rate of 0.914 m/sec (3 ft/sec) the system achieves a peak load reduction of 36% during landing impact.

Effects of short-term hypothermal and contrast exposure on immunophysiological parameters of laboratory animals.

PubMed

Kalenova, L F; Fisher, T A; Suhovey, J G; Besedin, I M

2009-05-01

Experiments on inbred animals showed that short-term exposure in cold water significantly modified structural and functional parameters of the immune system at different levels of its organization, from bone marrow hemopoiesis to effector stage of the immune response to antigen. The thermal factor caused changes in nonspecific and specific mechanisms of the immune system. Hypothermal exposure (7-9 degrees C, 5 sec) increased the thymic index and bone marrow lymphocyte count, reduced absorption capacity and stimulated metabolic activity of phagocytes, stimulated cell-mediated and suppressed humoral immunity. Contrast exposure in cold and hot water (7-9 degrees C, 5 sec/40-42 degrees C, 30 sec) increased monocyte count in bone marrow and reduced it in the their peripheral blood, reduced metabolic activity of phagocytes, stimulated cell-mediated and suppressed humoral immunity. These data demonstrate physiological mechanisms of interactions between the thermoregulatory and immune systems.

75 FR 8239 - Federal Home Loan Bank Housing Associates, Core Mission Activities and Standby Letters of Credit

Federal Register 2010, 2011, 2012, 2013, 2014

2010-02-24

... part 950 of this chapter. title. Sec. 1264.3(a), Sec. 926.4........ Sec. 1264.4. Introductory text. Sec...).. Sec. 1264.3(a)(2). Introductory text. Sec. 1264.4(c)(1)......... Sec. 926.3(a)(3).. Sec. 1264.3(a)(3...)......... Sec. 950.7 of this Sec. 950.7 of this chapter. title. Sec. 1269.3(a), Sec. Sec. Sec. Sec. Introductory...

Simulation study of two VTOL control/display systems in IMC approach and landing

NASA Technical Reports Server (NTRS)

Merrick, V. K.

1981-01-01

Both systems had full attitude command; the more complex system (Type 1) also had translational velocity command. The systems were applied to existing models of a VTOL lift-fan transport and the AV-8A Harrier. Simulated landings were made on a model of a DD963 Spruance-class destroyer. It was concluded that acceptable transitions and vertical landings can be performed, using the Type 1 system, in free-air turbulence up to 2.5 m/sec and sea state 6 and, using the Type 2 system, in free-air turbulence up to 1.5 m/sec and sea state 4.

Identification and characterization of a selenoprotein family containing a diselenide bond in a redox motif

PubMed Central

Shchedrina, Valentina A.; Novoselov, Sergey V.; Malinouski, Mikalai Yu.; Gladyshev, Vadim N.

2007-01-01

Selenocysteine (Sec, U) insertion into proteins is directed by translational recoding of specific UGA codons located upstream of a stem-loop structure known as Sec insertion sequence (SECIS) element. Selenoproteins with known functions are oxidoreductases containing a single redox-active Sec in their active sites. In this work, we identified a family of selenoproteins, designated SelL, containing two Sec separated by two other residues to form a UxxU motif. SelL proteins show an unusual occurrence, being present in diverse aquatic organisms, including fish, invertebrates, and marine bacteria. Both eukaryotic and bacterial SelL genes use single SECIS elements for insertion of two Sec. In eukaryotes, the SECIS is located in the 3′ UTR, whereas the bacterial SelL SECIS is within a coding region and positioned at a distance that supports the insertion of either of the two Sec or both of these residues. SelL proteins possess a thioredoxin-like fold wherein the UxxU motif corresponds to the catalytic CxxC motif in thioredoxins, suggesting a redox function of SelL proteins. Distantly related SelL-like proteins were also identified in a variety of organisms that had either one or both Sec replaced with Cys. Danio rerio SelL, transiently expressed in mammalian cells, incorporated two Sec and localized to the cytosol. In these cells, it occurred in an oxidized form and was not reducible by DTT. In a bacterial expression system, we directly demonstrated the formation of a diselenide bond between the two Sec, establishing it as the first diselenide bond found in a natural protein. PMID:17715293

Type-IVC Secretion System: A Novel Subclass of Type IV Secretion System (T4SS) Common Existing in Gram-Positive Genus Streptococcus

PubMed Central

Chen, Chen; Gao, George F.

2012-01-01

A growing number of pathogens are being found to possess specialized secretion systems which they use in various ways to subvert host defenses. Type IV secretion system (T4SS) is one of versatile secretion systems essential for the virulence and even survival of some bacteria species, and they enable the secretion of protein and DNA substrates across the cell envelope. T4SS was once believed to be present only in Gram-negative bacteria. In this study, we present evidence of a new subclass of T4SS, Type-IVC secretion system and indicate its common existence in the Gram-positive bacterial genus Streptococcus. We further identified that VirB1, VirB4, VirB6 and VirD4 are the minimal key components of this system. Using genome comparisons and evolutionary relationship analysis, we proposed that Type-IVC secretion system is movable via transposon factors and mediates the conjugative transfer of DNA, enhances bacterial pathogenicity, and could cause large-scale outbreaks of infections in humans. PMID:23056296

Analysis and design of ion thruster for large space systems

NASA Technical Reports Server (NTRS)

Poeschel, R. L.; Kami, S.

1980-01-01

Design analyses showed that an ion thruster of approximately 50 cm in diameter will be required to produce a thrust of 0.5 N using xenon or argon as propellants, and operating the thruster at a specific impulse of 3530 sec or 6076 sec respectively. A multipole magnetic confinement discharge chamber was specified.

Magnetostatic Field System for Uniform Cell Cultures Exposure

PubMed Central

Vergallo, Cristian; Piccoli, Claudia; Romano, Alberto; Panzarini, Elisa; Serra, Antonio; Manno, Daniela; Dini, Luciana

2013-01-01

The aim of the present work has been the design and the realization of a Magnetostatic Field System for Exposure of Cell cultures (MaFiSEC) for the uniform and the reproducible exposure of cell cultures to static magnetic fields (SMFs) of moderate magnetic induction. Experimental and computer-simulated physical measurements show that MaFiSEC: i) generates a SMF with magnetic induction that can be chosen in the range of 3 to 20 mT; ii) allows the uniform SMF exposure of cells growing in adhesion and in suspension; iii) is cheap and easy to use. The efficacy and reproducibility of MaFiSEC has been tested by comparing the biological effects exerted on isolated human lymphocytes by 72 h of exposure to a magnet (i.e. Neodymium Magnetic Disk, NMD) placed under the culture Petri dish. Lymphocytes morphology, viability, cell death, oxidative stress and lysosomes activity were the parameters chosen to evaluate the SMF biological effects. The continuous exposure of cells to a uniform SMF, achieved with MaFiSEC, allows highly reproducible biochemical and morphological data. PMID:23977284

Identification of protein components and quantitative immunoassay for SEC2 in staphylococcin injection.

PubMed

Ding, Ding; Huang, Peng; Sun, Hongying; Xue, Qiao; Pan, Yingqiu; Chen, Shuqing

2009-08-15

In China, staphylococcin injection has been commonly used in combined cancer therapy to enhance the systemic immune response and reduce the toxicities associated with chemotherapy or radiation therapy in the last decade. It is claimed that the main effective component is staphylococcal enterotoxin C2 (SEC2). However, no standard method based on the concentration of SEC2 has been established for quality control of the injection products. In this study, a sensitive and reliable biotin-streptavidin-ELISA (BS-ELISA) method was established for detection and quantification of SEC2. In addition, 1-D SDS-PAGE coupled with nano-LC-MS/MS was performed to identify the protein components in the injection products from one manufacturing company. The results of the BS-ELISA showed that SEC2 only accounted for less than 0.1% of the total protein in the injection products, and the nano-LC-MS/MS results showed that fifty-five proteins of Staphylococcus aureus were confidently identified in the injection solution. Seventeen out of these proteins, including SEC2, were well-known virulence factors. In addition, eighteen proteins of other Gram-positive bacteria were also confidently identified. Thus, the results indicated that SEC2 is of very low concentration in the injection products and the process of the injection preparation should be improved for health and safety consideration.

AtlA functions as a peptidoglycan lytic transglycosylase in the Neisseria gonorrhoeae type IV secretion system.

PubMed

Kohler, Petra L; Hamilton, Holly L; Cloud-Hansen, Karen; Dillard, Joseph P

2007-08-01

Type IV secretion systems require peptidoglycan lytic transglycosylases for efficient secretion, but the function of these enzymes is not clear. The type IV secretion system gene cluster of Neisseria gonorrhoeae encodes two peptidoglycan transglycosylase homologues. One, LtgX, is similar to peptidoglycan transglycosylases from other type IV secretion systems. The other, AtlA, is similar to endolysins from bacteriophages and is not similar to any described type IV secretion component. We characterized the enzymatic function of AtlA in order to examine its role in the type IV secretion system. Purified AtlA was found to degrade macromolecular peptidoglycan and to produce 1,6-anhydro peptidoglycan monomers, characteristic of lytic transglycosylase activity. We found that AtlA can functionally replace the lambda endolysin to lyse Escherichia coli. In contrast, a sensitive measure of lysis demonstrated that AtlA does not lyse gonococci expressing it or gonococci cocultured with an AtlA-expressing strain. The gonococcal type IV secretion system secretes DNA during growth. A deletion of ltgX or a substitution in the putative active site of AtlA severely decreased DNA secretion. These results indicate that AtlA and LtgX are actively involved in type IV secretion and that AtlA is not involved in lysis of gonococci to release DNA. This is the first demonstration that a type IV secretion peptidoglycanase has lytic transglycosylase activity. These data show that AtlA plays a role in type IV secretion of DNA that requires peptidoglycan breakdown without cell lysis.

High speed holographic digital recorder.

PubMed

Roberts, H N; Watkins, J W; Johnson, R H

1974-04-01

Concepts, feasibility experiments, and key component developments are described for a holographic digital record/reproduce system with the potential for 1.0 Gbit/sec rates and higher. Record rates of 500 Mbits/sec have been demonstrated with a ten-channel acoustooptic modulator array and a mode-locked, cavity-dumped argon-ion laser. Acoustooptic device technology has been advanced notably during the development of mode lockers, cavity dumpers, beam deflectors, and multichannel modulator arrays. The development of high speed multichannel photodetector arrays for the readout subsystem requires special attention. The feasibility of 1.0 Gbits/sec record rates has been demonstrated.

Autonomous Mission Operations

NASA Technical Reports Server (NTRS)

Frank, Jeremy; Spirkovska, Lilijana; McCann, Rob; Wang, Lui; Pohlkamp, Kara; Morin, Lee

2012-01-01

NASA's Advanced Exploration Systems Autonomous Mission Operations (AMO) project conducted an empirical investigation of the impact of time-delay on todays mission operations, and of the effect of processes and mission support tools designed to mitigate time-delay related impacts. Mission operation scenarios were designed for NASA's Deep Space Habitat (DSH), an analog spacecraft habitat, covering a range of activities including nominal objectives, DSH system failures, and crew medical emergencies. The scenarios were simulated at time-delay values representative of Lunar (1.2-5 sec), Near Earth Object (NEO) (50 sec) and Mars (300 sec) missions. Each combination of operational scenario and time-delay was tested in a Baseline configuration, designed to reflect present-day operations of the International Space Station, and a Mitigation configuration in which a variety of software tools, information displays, and crew-ground communications protocols were employed to assist both crews and Flight Control Team (FCT) members with the long-delay conditions. Preliminary findings indicate: 1) Workload of both crew members and FCT members generally increased along with increasing time delay. 2) Advanced procedure execution viewers, caution and warning tools, and communications protocols such as text messaging decreased the workload of both flight controllers and crew, and decreased the difficulty of coordinating activities. 3) Whereas crew workload ratings increased between 50 sec and 300 sec of time-delay in the Baseline configuration, workload ratings decreased (or remained flat) in the Mitigation configuration.

Marshall Space Flight Center's Tower Vector Magnetograph: Upgrades, Hardware, and Operations for the HESSI Mission

NASA Technical Reports Server (NTRS)

Adams, M. L.; Hagyard, M. J.; West, E. A.; Smith, J. E.; Whitaker, Ann F. (Technical Monitor)

2001-01-01

The Marshall Space Flight Center's (MSFC) solar group announces the successful upgrade of our tower vector magnetograph. In operation since 1973, the last major alterations to the system (which includes telescope, filter, polarizing optics, camera, and data acquisition computer) were made in 1982, when we upgraded from an SEC Vidicon camera to a CCD. In 1985, other changes were made which increased the field-of-view from 5 x 5 arc min (2.4 arc sec per pixel) to 6 x 6 arc min with a resolution of 2.81 arc sec. In 1989, the Apollo Telescope Mount H-alpha telescope was coaligned with the optics of the magnetograph. The most recent upgrades (year 2000), funded to support the High Energy Solar Spectroscopic Imager (HESSI) mission, have resulted in a pixel size of 0.64 arc sec over a 7 x 5.2 arc min field-of-view (binning 1x1). This poster describes the physical characteristics of the new system and compares spatial resolution, timing, and versatility with the old system. Finally, we provide a description of our Internet web site, which includes images of our most recent observations, and links to our data archives, as well as the history of magnetography at MSFC and education outreach pages.

Analytical linear energy transfer model including secondary particles: calculations along the central axis of the proton pencil beam

NASA Astrophysics Data System (ADS)

Marsolat, F.; De Marzi, L.; Pouzoulet, F.; Mazal, A.

2016-01-01

In proton therapy, the relative biological effectiveness (RBE) depends on various types of parameters such as linear energy transfer (LET). An analytical model for LET calculation exists (Wilkens’ model), but secondary particles are not included in this model. In the present study, we propose a correction factor, L sec, for Wilkens’ model in order to take into account the LET contributions of certain secondary particles. This study includes secondary protons and deuterons, since the effects of these two types of particles can be described by the same RBE-LET relationship. L sec was evaluated by Monte Carlo (MC) simulations using the GATE/GEANT4 platform and was defined by the ratio of the LET d distributions of all protons and deuterons and only primary protons. This method was applied to the innovative Pencil Beam Scanning (PBS) delivery systems and L sec was evaluated along the beam axis. This correction factor indicates the high contribution of secondary particles in the entrance region, with L sec values higher than 1.6 for a 220 MeV clinical pencil beam. MC simulations showed the impact of pencil beam parameters, such as mean initial energy, spot size, and depth in water, on L sec. The variation of L sec with these different parameters was integrated in a polynomial function of the L sec factor in order to obtain a model universally applicable to all PBS delivery systems. The validity of this correction factor applied to Wilkens’ model was verified along the beam axis of various pencil beams in comparison with MC simulations. A good agreement was obtained between the corrected analytical model and the MC calculations, with mean-LET deviations along the beam axis less than 0.05 keV μm-1. These results demonstrate the efficacy of our new correction of the existing LET model in order to take into account secondary protons and deuterons along the pencil beam axis.

Computational and Experimental Analysis of the Secretome of Methylococcus capsulatus (Bath)

PubMed Central

Indrelid, Stine; Mathiesen, Geir; Jacobsen, Morten; Lea, Tor; Kleiveland, Charlotte R.

2014-01-01

The Gram-negative methanotroph Methylococcus capsulatus (Bath) was recently demonstrated to abrogate inflammation in a murine model of inflammatory bowel disease, suggesting interactions with cells involved in maintaining mucosal homeostasis and emphasizing the importance of understanding the many properties of M. capsulatus. Secreted proteins determine how bacteria may interact with their environment, and a comprehensive knowledge of such proteins is therefore vital to understand bacterial physiology and behavior. The aim of this study was to systematically analyze protein secretion in M. capsulatus (Bath) by identifying the secretion systems present and the respective secreted substrates. Computational analysis revealed that in addition to previously recognized type II secretion systems and a type VII secretion system, a type Vb (two-partner) secretion system and putative type I secretion systems are present in M. capsulatus (Bath). In silico analysis suggests that the diverse secretion systems in M.capsulatus transport proteins likely to be involved in adhesion, colonization, nutrient acquisition and homeostasis maintenance. Results of the computational analysis was verified and extended by an experimental approach showing that in addition an uncharacterized protein and putative moonlighting proteins are released to the medium during exponential growth of M. capsulatus (Bath). PMID:25479164

77 FR 76139 - Self-Regulatory Organizations; NYSE Arca, Inc.; Notice of Filing and Immediate Effectiveness of...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-12-26

... happen because of a temporary systems outage, a communications issue between the electronic and floor... Commission's Internet comment form ( http://www.sec.gov/rules/sro.shtml ); or Send an email to rule-comments... Commission will post all comments on the Commission's Internet Web site ( http://www.sec.gov/rules/sro.shtml...

40 CFR 86.1319-90 - CVS calibration.

Code of Federal Regulations, 2010 CFR

2010-07-01

... with such a device.) The CVS calibration procedures are designed for use of a “metering venturi” type... series with the pump. (ii) The calculated flow rate, ft 3/min, (at pump inlet absolute pressure and... during test period N Revs ±1 Rev. Elapsed time for test period t sec. ±0.5 sec. (5) After the system has...

40 CFR 86.1319-90 - CVS calibration.

Code of Federal Regulations, 2013 CFR

2013-07-01

... with such a device.) The CVS calibration procedures are designed for use of a “metering venturi” type... series with the pump. (ii) The calculated flow rate, ft 3/min, (at pump inlet absolute pressure and... test period N Revs ±1 Rev. Elapsed time for test period t sec. ±0.5 sec. (5) After the system has been...

40 CFR 86.1319-90 - CVS calibration.

Code of Federal Regulations, 2011 CFR

2011-07-01

... with such a device.) The CVS calibration procedures are designed for use of a “metering venturi” type... series with the pump. (ii) The calculated flow rate, ft 3/min, (at pump inlet absolute pressure and... during test period N Revs ±1 Rev. Elapsed time for test period t sec. ±0.5 sec. (5) After the system has...

40 CFR 86.1319-90 - CVS calibration.

Code of Federal Regulations, 2012 CFR

2012-07-01

... with such a device.) The CVS calibration procedures are designed for use of a “metering venturi” type... series with the pump. (ii) The calculated flow rate, ft 3/min, (at pump inlet absolute pressure and... test period N Revs ±1 Rev. Elapsed time for test period t sec. ±0.5 sec. (5) After the system has been...

75 FR 62404 - Agency Information Collection Activities; Proposed Collection; Comment Request; Additional...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-10-08

... TEA final rule). The regulations in Sec. 330.14 state that OTC drug products introduced into the U.S... and extent'' application (TEA) to be submitted to us by any party for our consideration to include new conditions in the OTC drug monograph system. TEAs must provide evidence described in Sec. 330.14(c...

Global Stability and Control Analysis of Aircraft at High Angles-of-Attack.

DTIC Science & Technology

1977-06-30

500 deg/sec to 500 deg/sec and com - puting the eigenvalues of the linearized system around different equilibrium points for 6e = 2°. It is noticed...SYSTEMS, INC. 0" -114- 2 Ä»" • + + + + + _+ + + + + + + + + M I o o o a i i 8 8 in 8 8 XNXX * x x» x x x< x...history com - parisons show significant improvement in dynamic response and performance using the BACTM ARI gain, as opposed to nulling the rudder

SecG is required for antibiotic activities of Pseudomonas sp. YL23 against Erwinia amylovora and Dickeya chrysanthemi.

PubMed

Liu, Youzhou; Baird, Sonya M; Qiao, Junqing; Du, Yan; Lu, Shi-En

2015-05-01

Strain YL23 was isolated from soybean root tips and identified to be Pseudomonas sp. This strain showed broad-spectrum antibacterial activity against bacterial pathogens that are economically important in agriculture. To characterize the genes dedicated to antibacterial activities against microbial phytopathogens, a Tn5-mutation library of YL23 was constructed. Plate bioassays revealed that the mutant YL23-93 lost its antibacterial activities against Erwinia amylovora and Dickeya chrysanthemi as compared with its wild type strain. Genetic and sequencing analyses localized the transposon in a homolog of the secG gene in the mutant YL23-93. Constitutive expression plasmid pUCP26-secG was constructed and electroporated into the mutant YL23-93. Introduction of the plasmid pUCP26-secG restored antibacterial activities of the mutant YL23-93 to E. amylovora and D. chrysanthemi. As expected, empty plasmid pUCP26 could not complement the phenotype of the antibacterial activity in the mutant. Thus the secG gene, belonging to the Sec protein translocation system, is required for antibacterial activity of strain YL23 against E. amylovora and D. chrysanthemi. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Hydroxytyrosol and its complex forms (secoiridoids) modulate aorta and heart proteome in healthy rats: Potential cardio-protective effects.

PubMed

Catalán, Úrsula; Rubió, Laura; López de Las Hazas, Maria-Carmen; Herrero, Pol; Nadal, Pedro; Canela, Núria; Pedret, Anna; Motilva, Maria-José; Solà, Rosa

2016-10-01

Hydroxytyrosol (HT) is the major phenolic compound in virgin olive oil (VOO) in both free and complex forms (secoiridoids; SEC). Proteomics of cardiovascular tissues such as aorta or heart represents a promising tool to uncover the mechanisms of action of phenolic compounds in healthy animals. Twelve female Wistar rats were separated into three groups: a standard diet and two diets supplemented in phenolic compounds (HT and SEC) adjusted to 5 mg/kg/day during 21 days. Proteomic analyses of aorta and heart tissues were performed by nano-LC and MS. Ingenuity Pathway Analysis was used to generate interaction networks. HT or SEC modulated aorta and heart proteome compared to the standard diet. The top-scored networks were related to Cardiovascular System. HT and SEC downregulated proteins related to proliferation and migration of endothelial cells and occlusion of blood vessels in aorta and proteins related to heart failure in heart tissue. SEC showed higher fold change values compared to HT, attributed to higher concentration of HT detected in heart tissue. Changes at proteomic level in cardiovascular tissues may partially account for the underlying mechanisms of VOO phenols cardiovascular protection being the SEC effects higher than free HT. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Ranging and tracking system for proximity operations, phase 1

NASA Technical Reports Server (NTRS)

Udalov, S.; Nilsen, P. W.

1979-01-01

A study task is reported which is directed towards developing a conceptual design of a small, lightweight range and range rate radar sensor system to meet NASA's requirements for accurate short-range and velocity measurements in an orbital environment. Within the context of the requirements, the short range implies system operation at 0 m to 1850 m (6000 ft) and accurate implies a range measurement to within 1 sigma accuracy of 0.20 m (0.67 ft) and a range rate (velocity) measurement to within 1 sigma accuracy of 0.01 m/sec (0.033 ft/sec).

Development of the Dual Aerodynamic Nozzle Model for the NTF Semi-Span Model Support System

NASA Technical Reports Server (NTRS)

Jones, Greg S.; Milholen, William E., II; Goodliff, Scott L.

2011-01-01

The recent addition of a dual flow air delivery system to the NASA Langley National Transonic Facility was experimentally validated with a Dual Aerodynamic Nozzle semi-span model. This model utilized two Stratford calibration nozzles to characterize the weight flow system of the air delivery system. The weight flow boundaries for the air delivery system were identified at mildly cryogenic conditions to be 0.1 to 23 lbm/sec for the high flow leg and 0.1 to 9 lbm/sec for the low flow leg. Results from this test verified system performance and identified problems with the weight-flow metering system that required the vortex flow meters to be replaced at the end of the test.

Progress in Y-00 physical cipher for Giga bit/sec optical data communications (intensity modulation method)

NASA Astrophysics Data System (ADS)

Hirota, Osamu; Futami, Fumio

2014-10-01

To guarantee a security of Cloud Computing System is urgent problem. Although there are several threats in a security problem, the most serious problem is cyber attack against an optical fiber transmission among data centers. In such a network, an encryption scheme on Layer 1(physical layer) with an ultimately strong security, a small delay, and a very high speed should be employed, because a basic optical link is operated at 10 Gbit/sec/wavelength. We have developed a quantum noise randomied stream cipher so called Yuen- 2000 encryption scheme (Y-00) during a decade. This type of cipher is a completely new type random cipher in which ciphertext for a legitimate receiver and eavesdropper are different. This is a condition to break the Shannon limit in theory of cryptography. In addition, this scheme has a good balance on a security, a speed and a cost performance. To realize such an encryption, several modulation methods are candidates such as phase-modulation, intensity-modulation, quadrature amplitude modulation, and so on. Northwestern university group demonstrated a phase modulation system (α=η) in 2003. In 2005, we reported a demonstration of 1 Gbit/sec system based on intensity modulation scheme(ISK-Y00), and gave a design method for quadratic amplitude modulation (QAM-Y00) in 2005 and 2010. An intensity modulation scheme promises a real application to a secure fiber communication of current data centers. This paper presents a progress in quantum noise randomized stream cipher based on ISK-Y00, integrating our theoretical and experimental achievements in the past and recent 100 Gbit/sec(10Gbit/sec × 10 wavelengths) experiment.

The type III secretion system is involved in Escherichia coli K1 interactions with Acanthamoeba.

PubMed

Siddiqui, Ruqaiyyah; Malik, Huma; Sagheer, Mehwish; Jung, Suk-Yul; Khan, Naveed Ahmed

2011-08-01

The type III secretion system among Gram-negative bacteria is known to deliver effectors into host cell to interfere with host cellular processes. The type III secretion system in Yersina, Pseudomonas and Enterohemorrhagic Escherichia coli have been well documented to be involved in the bacterial pathogenicity. The existence of type III secretion system has been demonstrated in neuropathogenic E. coli K1 strains. Here, it is observed that the deletion mutant of type III secretion system in E. coli strain EC10 exhibited defects in the invasion and intracellular survival in Acanthamoeba castellanii (a keratitis isolate) compared to its parent strain. Next, it was determined whether type III secretion system plays a role in E. coli K1 survival inside Acanthamoeba during the encystment process. Using encystment assays, our findings revealed that the type III secretion system-deletion mutant exhibited significantly reduced survival inside Acanthamoeba cysts compared with its parent strain, EC10 (P<0.01). This is the first demonstration that the type III secretion system plays an important role in E. coli interactions with Acanthamoeba. A complete understanding of how amoebae harbor bacterial pathogens will help design strategies against E. coli transmission to the susceptible hosts. Copyright © 2011 Elsevier Inc. All rights reserved.

Distribution and diversity of bacterial secretion systems across metagenomic datasets.

PubMed

Barret, Matthieu; Egan, Frank; O'Gara, Fergal

2013-02-01

Bacteria can manipulate their surrounding environment through the secretion of proteins into other living organisms and into the extracellular milieu. In Gram stain negative bacteria this process is mediated by different types of secretion systems from type I through type VI secretion system (T1SS-T6SS). In this study the prevalence of these secretion systems in 312 publicly available microbiomes derived from a wide range of ecosystems was investigated by a gene-centric approach. Our analysis demonstrates that some secretion systems are over-represented in some specific samples. In addition, some T3SS and T6SS phylogenetic clusters were specifically enriched in particular ecological niches, which could indicate specific bacterial adaptation to these environments. © 2012 Society for Applied Microbiology and Blackwell Publishing Ltd.

An RNAi Screen for Genes Involved in Nanoscale Protrusion Formation on Corneal Lens in Drosophila melanogaster.

PubMed

Minami, Ryunosuke; Sato, Chiaki; Yamahama, Yumi; Kubo, Hideo; Hariyama, Takahiko; Kimura, Ken-Ichi

2016-12-01

The "moth-eye" structure, which is observed on the surface of corneal lens in several insects, supports anti-reflective and self-cleaning functions due to nanoscale protrusions known as corneal nipples. Although the morphology and function of the "moth-eye" structure, are relatively well studied, the mechanism of protrusion formation from cell-secreted substances is unknown. In Drosophila melanogaster, a compound eye consists of approximately 800 facets, the surface of which is formed by the corneal lens with nanoscale protrusions. In the present study, we sought to identify genes involved in "moth-eye" structure, formation in order to elucidate the developmental mechanism of the protrusions in Drosophila. We re-examined the aberrant patterns in classical glossy-eye mutants by scanning electron microscope and classified the aberrant patterns into groups. Next, we screened genes encoding putative structural cuticular proteins and genes involved in cuticular formation using eye specific RNAi silencing methods combined with the Gal4/UAS expression system. We identified 12 of 100 candidate genes, such as cuticular proteins family genes (Cuticular protein 23B and Cuticular protein 49Ah), cuticle secretion-related genes (Syntaxin 1A and Sec61 ββ subunit), ecdysone signaling and biosynthesis-related genes (Ecdysone receptor, Blimp-1, and shroud), and genes involved in cell polarity/cell architecture (Actin 5C, shotgun, armadillo, discs large1, and coracle). Although some of the genes we identified may affect corneal protrusion formation indirectly through general patterning defects in eye formation, these initial findings have encouraged us to more systematically explore the precise mechanisms underlying the formation of nanoscale protrusions in Drosophila.

Biosynthesis of Selenocysteine on Its tRNA in Eukaryotes

PubMed Central

Mix, Heiko; Zhang, Yan; Saira, Kazima; Glass, Richard S; Berry, Marla J; Gladyshev, Vadim N; Hatfield, Dolph L

2007-01-01

Selenocysteine (Sec) is cotranslationally inserted into protein in response to UGA codons and is the 21st amino acid in the genetic code. However, the means by which Sec is synthesized in eukaryotes is not known. Herein, comparative genomics and experimental analyses revealed that the mammalian Sec synthase (SecS) is the previously identified pyridoxal phosphate-containing protein known as the soluble liver antigen. SecS required selenophosphate and O-phosphoseryl-tRNA[Ser]Sec as substrates to generate selenocysteyl-tRNA[Ser]Sec. Moreover, it was found that Sec was synthesized on the tRNA scaffold from selenide, ATP, and serine using tRNA[Ser]Sec, seryl-tRNA synthetase, O-phosphoseryl-tRNA[Ser]Sec kinase, selenophosphate synthetase, and SecS. By identifying the pathway of Sec biosynthesis in mammals, this study not only functionally characterized SecS but also assigned the function of the O-phosphoseryl-tRNA[Ser]Sec kinase. In addition, we found that selenophosphate synthetase 2 could synthesize monoselenophosphate in vitro but selenophosphate synthetase 1 could not. Conservation of the overall pathway of Sec biosynthesis suggests that this pathway is also active in other eukaryotes and archaea that synthesize selenoproteins. PMID:17194211

75 FR 16222 - Self-Regulatory Organizations; Notice of Filing and Immediate Effectiveness of Proposed Rule...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-03-31

... liability for system outages. The text of the proposed rule change is attached as Exhibit 5 to the 19b-4...: Electronic Comments Use the Commission's Internet comment form ( http://www.sec.gov/rules/sro.shtml ); or... method. The Commission will post all comments on the Commission's Internet Web site ( http://www.sec.gov...

Comparison of DMSP and SECS region-1 and region-2 ionospheric current boundary

NASA Astrophysics Data System (ADS)

Weygand, J. M.; Wing, S.

2016-06-01

The region-1 and region-2 boundary has traditionally been identified using data from a single spacecraft crossing the auroral region and measuring the large scale changes in the cross track magnetic field. With data from the AUTUMN, CANMOS, CARISMA, GIMA, DTU MGS, MACCS, McMAC, STEP, THEMIS, and USGS ground magnetometer arrays we applied a state-of-art technique based on spherical elementary current system (SECS) method developed by Amm and Viljanen (1999) in order to calculate maps of region-1 and region-2 current system over the North American and Greenland auroral region. Spherical elementary current (SEC) amplitude (proxy for vertical currents) maps can be inferred at 10 s temporal resolution, ~1.5° geographic latitude (Glat), and 3.5° geographic longitude (Glon) spatial resolution. We compare the location of the region-1 and region-2 boundary obtained by the DMSP spacecraft with the region-1 and region-2 boundary observed in the SEC current amplitudes. We find that the boundaries typically agree within 0.2°±1.3°. These results indicate that the location of the region-1 and region-2 boundary can reasonably be determined from ground magnetometer data. The SECS maps represent a value-added product from the magnetometer database and can be used for contextual interpretation in conjunction with other missions as well as help with our understanding of magnetosphere-ionosphere coupling mechanisms using the ground arrays and the magnetospheric spacecraft data.

Comparison of DMSP and SECS region-1 and region-2 ionospheric current boundary✩

PubMed Central

Weygand, J.M.; Wing, S.

2017-01-01

The region-1 and region-2 boundary has traditionally been identified using data from a single spacecraft crossing the auroral region and measuring the large scale changes in the cross track magnetic field. With data from the AUTUMN, CANMOS, CARISMA, GIMA, DTU MGS, MACCS, McMAC, STEP, THEMIS, and USGS ground magnetometer arrays we applied a state-of-art technique based on spherical elementary current system (SECS) method developed by Amm and Viljanen (1999) in order to calculate maps of region-1 and region-2 current system over the North American and Greenland auroral region. Spherical elementary current (SEC) amplitude (proxy for vertical currents) maps can be inferred at 10 s temporal resolution, ~1.5° geographic latitude (Glat), and 3.5° geographic longitude (Glon) spatial resolution. We compare the location of the region-1 and region-2 boundary obtained by the DMSP spacecraft with the region-1 and region-2 boundary observed in the SEC current amplitudes. We find that the boundaries typically agree within 0.2° ± 1.3°. These results indicate that the location of the region-1 and region-2 boundary can reasonably be determined from ground magnetometer data. The SECS maps represent a value-added product from the magnetometer database and can be used for contextual interpretation in conjunction with other missions as well as help with our understanding of magnetosphere-ionosphere coupling mechanisms using the ground arrays and the magnetospheric spacecraft data. PMID:29056861

Comparison of DMSP and SECS region-1 and region-2 ionospheric current boundary.

PubMed

Weygand, J M; Wing, S

2016-06-01

The region-1 and region-2 boundary has traditionally been identified using data from a single spacecraft crossing the auroral region and measuring the large scale changes in the cross track magnetic field. With data from the AUTUMN, CANMOS, CARISMA, GIMA, DTU MGS, MACCS, McMAC, STEP, THEMIS, and USGS ground magnetometer arrays we applied a state-of-art technique based on spherical elementary current system (SECS) method developed by Amm and Viljanen (1999) in order to calculate maps of region-1 and region-2 current system over the North American and Greenland auroral region. Spherical elementary current (SEC) amplitude (proxy for vertical currents) maps can be inferred at 10 s temporal resolution, ~1.5° geographic latitude (Glat), and 3.5° geographic longitude (Glon) spatial resolution. We compare the location of the region-1 and region-2 boundary obtained by the DMSP spacecraft with the region-1 and region-2 boundary observed in the SEC current amplitudes. We find that the boundaries typically agree within 0.2° ± 1.3°. These results indicate that the location of the region-1 and region-2 boundary can reasonably be determined from ground magnetometer data. The SECS maps represent a value-added product from the magnetometer database and can be used for contextual interpretation in conjunction with other missions as well as help with our understanding of magnetosphere-ionosphere coupling mechanisms using the ground arrays and the magnetospheric spacecraft data.

Piloted simulator study of allowable time delays in large-airplane response

NASA Technical Reports Server (NTRS)

Grantham, William D.; Bert T.?aetingas, Stephen A.dings with ran; Bert T.?aetingas, Stephen A.dings with ran

1987-01-01

A piloted simulation was performed to determine the permissible time delay and phase shift in the flight control system of a specific large transport-type airplane. The study was conducted with a six degree of freedom ground-based simulator and a math model similar to an advanced wide-body jet transport. Time delays in discrete and lagged form were incorporated into the longitudinal, lateral, and directional control systems of the airplane. Three experienced pilots flew simulated approaches and landings with random localizer and glide slope offsets during instrument tracking as their principal evaluation task. Results of the present study suggest a level 1 (satisfactory) handling qualities limit for the effective time delay of 0.15 sec in both the pitch and roll axes, as opposed to a 0.10-sec limit of the present specification (MIL-F-8785C) for both axes. Also, the present results suggest a level 2 (acceptable but unsatisfactory) handling qualities limit for an effective time delay of 0.82 sec and 0.57 sec for the pitch and roll axes, respectively, as opposed to 0.20 sec of the present specifications for both axes. In the area of phase shift between cockpit input and control surface deflection,the results of this study, flown in turbulent air, suggest less severe phase shift limitations for the approach and landing task-approximately 50 deg. in pitch and 40 deg. in roll - as opposed to 15 deg. of the present specifications for both axes.

Wind estimates from cloud motions: Phase 1 of an in situ aircraft verification experiment

NASA Technical Reports Server (NTRS)

Hasler, A. F.; Shenk, W. E.; Skillman, W.

1974-01-01

An initial experiment was conducted to verify geostationary satellite derived cloud motion wind estimates with in situ aircraft wind velocity measurements. Case histories of one-half hour to two hours were obtained for 3-10km diameter cumulus cloud systems on 6 days. Also, one cirrus cloud case was obtained. In most cases the clouds were discrete enough that both the cloud motion and the ambient wind could be measured with the same aircraft Inertial Navigation System (INS). Since the INS drift error is the same for both the cloud motion and wind measurements, the drift error subtracts out of the relative motion determinations. The magnitude of the vector difference between the cloud motion and the ambient wind at the cloud base averaged 1.2 m/sec. The wind vector at higher levels in the cloud layer differed by about 3 m/sec to 5 m/sec from the cloud motion vector.

Macrophage and tumor cell responses to repetitive pulsed X-ray radiation

NASA Astrophysics Data System (ADS)

Buldakov, M. A.; Tretyakova, M. S.; Ryabov, V. B.; Klimov, I. A.; Kutenkov, O. P.; Kzhyshkowska, J.; Bol'shakov, M. A.; Rostov, V. V.; Cherdyntseva, N. V.

2017-05-01

To study a response of tumor cells and macrophages to the repetitive pulsed low-dose X-ray radiation. Methods. Tumor growth and lung metastasis of mice with an injected Lewis lung carcinoma were analysed, using C57Bl6. Monocytes were isolated from a human blood, using CD14+ magnetic beads. IL6, IL1-betta, and TNF-alpha were determined by ELISA. For macrophage phenotyping, a confocal microscopy was applied. “Sinus-150” was used for the generation of pulsed X-ray radiation (the absorbed dose was below 0.1 Gy, the pulse repetition frequency was 10 pulse/sec). The irradiation of mice by 0.1 Gy pulsed X-rays significantly inhibited the growth of primary tumor and reduced the number of metastatic colonies in the lung. Furthermore, the changes in macrophage phenotype and cytokine secretion were observed after repetitive pulsed X-ray radiation. Conclusion. Macrophages and tumor cells had a different response to a low-dose pulsed X-ray radiation. An activation of the immune system through changes of a macrophage phenotype can result in a significant antitumor effect of the low-dose repetitive pulsed X-ray radiation.

Selenium. Role of the Essential Metalloid in Health

PubMed Central

Kurokawa, Suguru; Berry, Marla J.

2015-01-01

Selenium is an essential micronutrient in mammals, but is also recognized as toxic in excess. It is a non-metal with properties that are intermediate between the chalcogen elements sulfur and tellurium. Selenium exerts its biological functions through selenoproteins. Selenoproteins contain selenium in the form of the 21st amino acid, selenocysteine (Sec), which is an analog of cysteine with the sulfur-containing side chain replaced by a Se-containing side chain. Sec is encoded by the codon UGA, which is one of three termination codons for mRNA translation in non-selenoprotein genes. Recognition of the UGA codon as a Sec insertion site instead of stop requires a Sec insertion sequence (SECIS) element in selenoprotein mRNAs and a unique selenocysteyl-tRNA, both of which are recognized by specialized protein factors. Unlike the 20 standard amino acids, Sec is biosynthesized from serine on its tRNA. Twenty-five selenoproteins are encoded in the human genome. Most of the selenoprotein genes were discovered by bioinformatics approaches, searching for SECIS elements downstream of in-frame UGA codons. Sec has been described as having stronger nucleophilic and electrophilic properties than cysteine, and Sec is present in the catalytic site of all selenoenzymes. Most selenoproteins, whose functions are known, are involved in redox systems and signaling pathways. However, several selenoproteins are not well characterized in terms of their function. The selenium field has grown dramatically in the last few decades, and research on selenium biology is providing extensive new information regarding its importance for human health. PMID:24470102

SecPop Version 4: Sector Population Land Fraction and Economic Estimation Program: Users? Guide Model Manual and Verification Report.

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

Weber, Scott; Bixler, Nathan E.; McFadden, Katherine Letizia

In 1973 the U.S. Environmental Protection Agency (EPA) developed SecPop to calculate population estimates to support a study on air quality. The Nuclear Regulatory Commission (NRC) adopted this program to support siting reviews for nuclear power plant construction and license applications. Currently SecPop is used to prepare site data input files for offsite consequence calculations with the MELCOR Accident Consequence Code System (MACCS). SecPop enables the use of site-specific population, land use, and economic data for a polar grid defined by the user. Updated versions of SecPop have been released to use U.S. decennial census population data. SECPOP90 was releasedmore » in 1997 to use 1990 population and economic data. SECPOP2000 was released in 2003 to use 2000 population data and 1997 economic data. This report describes the current code version, SecPop version 4.3, which uses 2010 population data and both 2007 and 2012 economic data. It is also compatible with 2000 census and 2002 economic data. At the time of this writing, the current version of SecPop is 4.3.0, and that version is described herein. This report contains guidance for the installation and use of the code as well as a description of the theory, models, and algorithms involved. This report contains appendices which describe the development of the 2010 census file, 2007 county file, and 2012 county file. Finally, an appendix is included that describes the validation assessments performed.« less

Femtosecond Dynamics of the Photo-Induced Lattice Rearrangements in Quasi-One Halogen-Bridged Platinum Complexes

NASA Astrophysics Data System (ADS)

Suemoto, Tohru; Tomimoto, Shinichi; Matsuoka, Taira

Recent developments in femtosecond dynamics of the photoexcited state in quasi-one-dimensional platinum complexes [Pt(en)2][Pt(en)2X2] (ClO4)4 with X = Cl, Br and I are reviewed. The experimental results of time-resolved luminescence spectroscopy based on up-conversion technique are presented and analyzed in terms of a theory of wave-packet motion. An attempt to make a movie of wave-packet motion is mentioned. In Sec. 1, a brief introduction to the dynamics of the excited states in quasi-one-dimensional platinum complexes is given. It is stressed that this system can be a good model system for investigating the photo-induced structural phase transition. In order to describe a one-dimensional chain consisting of metal ions and halogen ions, the extended Peierls-Hubbard model is introduced in Sec. 2. The theoretical model of the relaxation dynamics in the excited states with a strong electron-lattice coupling is given in Sec. 3. The model is based on the interaction mode, which is appropriate for understanding the vibrational relaxation of localized centers in solids. Experimental backgrounds with some historical survey are given in Sec. 4. The recent experimental results of time-resolved luminescence for Pt-Cl, Pt-Br and Pt-I systems are presented in Secs. 5 to 8. The main result contains the direct observation of the wave-packet oscillation in the self-trapped excitons. The relaxation process observed in experiments has been successfully interpreted in terms of the model based on the interaction mode and the dynamical aspects are compared with the transient absorption measurements. The lifetime of the STE is shorter in Pt-X with heavier halogen ions. This behavior is discussed in relation with the non-radiative process leading to lattice rearrangements. In Secs. 9 and 10, visualization of the wave-packet form is presented. The basic behavior of the wave-packet is well understood in terms of a harmonic oscillator model. A non-exponential decay profiles are revealed from the center of gravity motion of the wave-packets. The exciton localization process is also discussed in the last section.

Ultra-high performance size-exclusion chromatography in polar solvents.

PubMed

Vancoillie, Gertjan; Vergaelen, Maarten; Hoogenboom, Richard

2016-12-23

Size-exclusion chromatography (SEC) is amongst the most widely used polymer characterization methods in both academic and industrial polymer research allowing the determination of molecular weight and distribution parameters, i.e. the dispersity (Ɖ), of unknown polymers. The many advantages, including accuracy, reproducibility and low sample consumption, have contributed to the worldwide success of this analytical technique. The current generation of SEC systems have a stationary phase mostly containing highly porous, styrene-divinylbenzene particles allowing for a size-based separation of various polymers in solution but limiting the flow rate and solvent compatibility. Recently, sub-2μm ethylene-bridged hybrid (BEH) packing materials have become available for SEC analysis. These packing materials can not only withstand much higher pressures up to 15000psi but also show high spatial stability towards different solvents. Combining these BEH columns with the ultra-high performance LC (UHPLC) technology opens up UHP-SEC analysis, showing strongly reduced runtimes and unprecedented solvent compatibility. In this work, this novel characterization technique was compared to conventional SEC using both highly viscous and highly polar solvents as eluent, namely N,N-dimethylacetamide (DMAc), N,N-dimethylformamide (DMF) and methanol, focusing on the suitability of the BEH-columns for analysis of highly functional polymers. The results show a high functional group compatibility comparable with conventional SEC with remarkably short runtimes and enhanced resolution in methanol. Copyright © 2016 Elsevier B.V. All rights reserved.

H.R. 3627: This Act may be cited as the Brownfield Community Empowerment Act, introduced in the House of Representatives, One Hundred Fifth Congress, Second Session, April 1, 1998

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

NONE

1998-12-31

The purpose of this bill is to assist local governments and local citizens` organizations in the assessment and remediation of brownfield sites, and for other purposes. Attention is focused on the following: Sec. 101. Inventory, assessment, and training and grant program; Sec. 112. Grants for revolving loan programs; Sec. 113. Economic redevelopment grants; Sec. 114. Reports; Sec 115. Limitations on use of funds; Sec. 116 Siting of TSD facilities; Sec. 117. Effect on other laws; Sec. 118. Regulations; Sec. 119. Authorizations of appropriations; Sec 120. Research, development, and demonstration; Sec. 121. Assistance for workforce training; Sec. 122. Worker training andmore » education grants; and Sec. 201. Economic development grants in connection with community development loan guarantees.« less

The enteric nervous system modulates mammalian duodenal mucosal bicarbonate secretion.

PubMed

Hogan, D L; Yao, B; Steinbach, J H; Isenberg, J I

1993-08-01

Interaction of the enteric nerves in regulating mammalian duodenal mucosal bicarbonate secretion is not well understood. The purpose of the present experiments was to evaluate the role of the enteric nervous system on bicarbonate secretion from rabbit duodenal mucosa in vitro. Proximal duodenum from male New Zealand White rabbits was stripped of seromuscular layers, mounted in Ussing chambers, and studied under short-circuited conditions. Effects of electrical field stimulation, vasoactive intestinal polypeptide (VIP), carbachol, prostaglandin E2 (PGE2), dibutyryl-cyclic adenosine monophosphate (db-cAMP), and the neurotoxin tetrodotoxin (TTX) and muscarinic blockade by atropine were studied. Electrical field stimulation significantly (P < 0.01) stimulated bicarbonate secretion, short-circuit current (Isc), and electrical potential difference (PD) that was sensitive to both TTX and atropine. VIP-stimulated bicarbonate secretion was significantly inhibited by TTX (-73%), yet Isc and PD remained unchanged. Atropine decreased VIP-induced bicarbonate secretion (-69%) and Isc (-43%). Carbachol-stimulated bicarbonate secretion, Isc, and PD were abolished by atropine, whereas TTX was without affect. Neither TTX nor atropine had a significant effect on PGE2 or db-cAMP-stimulated bicarbonate secretion. These results suggest that (1) enteric nerve stimulation activates an acetylcholine receptor that in turn stimulates duodenal epithelial bicarbonate secretion; (2) VIP stimulates bicarbonate secretion, in large part, via the enteric nervous system; and (3) PGE2 and cAMP stimulate bicarbonate secretion independent of the enteric nervous system.

The BOS1 gene encodes an essential 27-kD putative membrane protein that is required for vesicular transport from the ER to the Golgi complex in yeast

PubMed Central

1991-01-01

We recently described the identification of BOS1 (Newman, A., J. Shim, and S. Ferro-Novick. 1990. Mol. Cell. Biol. 10:3405-3414.). BOS1 is a gene that in multiple copy suppresses the growth and secretion defect of bet1 and sec22, two mutants that disrupt transport from the ER to the Golgi complex in yeast. The ability of BOS1 to specifically suppress mutants blocked at a particular stage of the secretory pathway suggested that this gene encodes a protein that functions in this process. The experiments presented in this study support this hypothesis. Specifically, the BOS1 gene was found to be essential for cellular growth. Furthermore, cells depleted of the Bos1 protein fail to transport pro-alpha-factor and carboxypeptidase Y (CPY) to the Golgi apparatus. This defect in export leads to the accumulation of an extensive network of ER and small vesicles. DNA sequence analysis predicts that Bos1 is a 27-kD protein containing a putative membrane- spanning domain. This prediction is supported by differential centrifugation experiments. Thus, Bos1 appears to be a membrane protein that functions in conjunction with Bet1 and Sec22 to facilitate the transport of proteins at a step subsequent to translocation into the ER but before entry into the Golgi apparatus. PMID:2007627

Proposed fast-response oxygen monitoring and control system for the Langley 8-foot high-temperature tunnel

NASA Technical Reports Server (NTRS)

Singh, J. J.; Davis, W. T.; Puster, R. L.

1983-01-01

A fast-response oxygen monitoring and control system, based on a Y2O3-stabilized ZrO2 sensor, was developed and tested in the laboratory. The system is capable of maintaining oxygen concentration in the CH4-O2-air combustion product gases at 20.9 + or - 1.0 percent. If the oxygen concentration in the exhaust stream differs from that in normal air by 25 percent or more, an alarm signal is provided for automatic tunnel shutdown. The overall prototype system response time was reduced from about 1 sec in the original configuration to about 0.2 sec. The basis of operation and the results of laboratory tests of the system are described.

Protein Secretion Systems in Pseudomonas aeruginosa: An Essay on Diversity, Evolution, and Function

PubMed Central

Filloux, Alain

2011-01-01

Protein secretion systems are molecular nanomachines used by Gram-negative bacteria to thrive within their environment. They are used to release enzymes that hydrolyze complex carbon sources into usable compounds, or to release proteins that capture essential ions such as iron. They are also used to colonize and survive within eukaryotic hosts, causing acute or chronic infections, subverting the host cell response and escaping the immune system. In this article, the opportunistic human pathogen Pseudomonas aeruginosa is used as a model to review the diversity of secretion systems that bacteria have evolved to achieve these goals. This diversity may result from a progressive transformation of cell envelope complexes that initially may not have been dedicated to secretion. The striking similarities between secretion systems and type IV pili, flagella, bacteriophage tail, or efflux pumps is a nice illustration of this evolution. Differences are also needed since various secretion configurations call for diversity. For example, some proteins are released in the extracellular medium while others are directly injected into the cytosol of eukaryotic cells. Some proteins are folded before being released and transit into the periplasm. Other proteins cross the whole cell envelope at once in an unfolded state. However, the secretion system requires conserved basic elements or features. For example, there is a need for an energy source or for an outer membrane channel. The structure of this review is thus quite unconventional. Instead of listing secretion types one after each other, it presents a melting pot of concepts indicating that secretion types are in constant evolution and use basic principles. In other words, emergence of new secretion systems could be predicted the way Mendeleïev had anticipated characteristics of yet unknown elements. PMID:21811488

Ultrahigh pressure fast size exclusion chromatography for top-down proteomics.

PubMed

Chen, Xin; Ge, Ying

2013-09-01

Top-down MS-based proteomics has gained a solid growth over the past few years but still faces significant challenges in the LC separation of intact proteins. In top-down proteomics, it is essential to separate the high mass proteins from the low mass species due to the exponential decay in S/N as a function of increasing molecular mass. SEC is a favored LC method for size-based separation of proteins but suffers from notoriously low resolution and detrimental dilution. Herein, we reported the use of ultrahigh pressure (UHP) SEC for rapid and high-resolution separation of intact proteins for top-down proteomics. Fast separation of intact proteins (6-669 kDa) was achieved in < 7 min with high resolution and high efficiency. More importantly, we have shown that this UHP-SEC provides high-resolution separation of intact proteins using a MS-friendly volatile solvent system, allowing the direct top-down MS analysis of SEC-eluted proteins without an additional desalting step. Taken together, we have demonstrated that UHP-SEC is an attractive LC strategy for the size separation of proteins with great potential for top-down proteomics. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Analysis of switch and examine combining with post-examining selection in cognitive radio

NASA Astrophysics Data System (ADS)

Agarwal, Rupali; Srivastava, Neelam; Katiyar, Himanshu

2018-06-01

To perform spectrum sensing in fading environment is one of the most challenging tasks for a CR system. Diversity combining schemes are used to combat the effect of fading and hence detection probability of CR gets improved. Among many diversity combining techniques, switched diversity offers one of the lowest complexity solutions. The receiver embedded with switched diversity looks for an acceptable diversity path (having signal to noise ratio (SNR) above the required threshold) to receive the data. In conventional switch and examine combining (SEC) scheme, when no acceptable path is found after all the paths are examined, the receiver randomly chooses an unacceptable path. Switch and examine combining with post-examining selection (SECp) is a modified version of conventional SEC. In SECp, the conventional SEC scheme is altered in a way that it selects the best path when no acceptable path is found after all paths have been examined. In this paper, formula for probability of detection ( ?) is derived using SECp and SEC diversity combining technique over Rayleigh fading channel. Also the performance of SECp is compared with SEC and no diversity case. Performance comparison is done with the help of SNR vs. ? and complementary receiver operating characteristic curves.

Mitigating the effects of landscape development on streams in urbanizing watersheds

USGS Publications Warehouse

Hogan, Dianna M.; Jarnagin, S. Taylor; Loperfido, John V.; Van Ness, Keith

2013-01-01

This collaborative study examined urbanization and impacts on area streams while using the best available sediment and erosion control (S&EC) practices in developing watersheds in Maryland, United States. During conversion of the agricultural and forested watersheds to urban land use, land surface topography was graded and vegetation was removed creating a high potential for sediment generation and release during storm events. The currently best available S&EC facilities were used during the development process to mitigate storm runoff water quality, quantity, and timing before entering area streams. Detailed Geographic Information System (GIS) maps were created to visualize changing land use and S&EC practices, five temporal collections of LiDAR (light detection and ranging) imagery were used to map the changing landscape topography, and streamflow, physical geomorphology, and habitat data were used to assess the ability of the S&EC facilities to protect receiving streams during development. Despite the use of the best available S&EC facilities, receiving streams experienced altered flow, geomorphology, and decreased biotic community health. These impacts on small streams during watershed development affect sediment and nutrient loads to larger downstream aquatic ecosystems such as the Chesapeake Bay.

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

Lou, K; Rice University, Houston, TX; Sun, X

Purpose: To study the feasibility of clinical on-line proton beam range verification with PET imaging Methods: We simulated a 179.2-MeV proton beam with 5-mm diameter irradiating a PMMA phantom of human brain size, which was then imaged by a brain PET with 300*300*100-mm{sup 3} FOV and different system sensitivities and spatial resolutions. We calculated the mean and standard deviation of positron activity range (AR) from reconstructed PET images, with respect to different data acquisition times (from 5 sec to 300 sec with 5-sec step). We also developed a technique, “Smoothed Maximum Value (SMV)”, to improve AR measurement under a givenmore » dose. Furthermore, we simulated a human brain irradiated by a 110-MeV proton beam of 50-mm diameter with 0.3-Gy dose at Bragg peak and imaged by the above PET system with 40% system sensitivity at the center of FOV and 1.7-mm spatial resolution. Results: MC Simulations on the PMMA phantom showed that, regardless of PET system sensitivities and spatial resolutions, the accuracy and precision of AR were proportional to the reciprocal of the square root of image count if image smoothing was not applied. With image smoothing or SMV method, the accuracy and precision could be substantially improved. For a cylindrical PMMA phantom (200 mm diameter and 290 mm long), the accuracy and precision of AR measurement could reach 1.0 and 1.7 mm, with 100-sec data acquired by the brain PET. The study with a human brain showed it was feasible to achieve sub-millimeter accuracy and precision of AR measurement with acquisition time within 60 sec. Conclusion: This study established the relationship between count statistics and the accuracy and precision of activity-range verification. It showed the feasibility of clinical on-line BR verification with high-performance PET systems and improved AR measurement techniques. Cancer Prevention and Research Institute of Texas grant RP120326, NIH grant R21CA187717, The Cancer Center Support (Core) Grant CA016672 to MD Anderson Cancer Center.« less

Identification of genes encoding the type IX secretion system and secreted proteins in Flavobacterium columnare IA-S-4

USDA-ARS?s Scientific Manuscript database

Flavobacterium columnare, a member of the phylum Bacteroidetes, causes columnaris disease in wild and aquaculture-reared freshwater fish. The mechanisms responsible for columnaris disease are not known. Many members of the phylum Bacteroidetes use type IX secretion systems (T9SSs) to secrete enzymes...

IR spectroscopy analysis of pancreatic lipase-related protein 2 interaction with phospholipids: 2. Discriminative recognition of various micellar systems and characterization of PLRP2-DPPC-bile salt complexes.

PubMed

Mateos-Diaz, Eduardo; Sutto-Ortiz, Priscila; Sahaka, Moulay; Byrne, Deborah; Gaussier, Hélène; Carrière, Frédéric

2018-03-01

The interaction of pancreatic lipase-related protein 2 (PLRP2) with various micelles containing phospholipids was investigated using pHstat enzyme activity measurements, differential light scattering, size exclusion chromatography (SEC) and transmission IR spectroscopy. Various micelles of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and lysophosphatidylcholine were prepared with either bile salts (sodium taurodeoxycholate or glycodeoxycholate) or Triton X-100, which are substrate-dispersing agents commonly used for measuring phospholipase activities. PLRP2 displayed a high activity on all phospholipid-bile salt micelles, but was totally inactive on phospholipid-Triton X-100 micelles. These findings clearly differentiate PLRP2 from secreted pancreatic phospholipase A2 which is highly active on both types of micelles. Using an inactive variant of PLRP2, SEC experiments allowed identifying two populations of PLRP2-DPPC-bile salt complexes corresponding to a high molecular weight 1:1 PLRP2-micelle association and to a low molecular weight association of PLRP2 with few monomers of DPPC/bile salts. IR spectroscopy analysis showed how DPPC-bile salt micelles differ from DPPC-Triton X-100 micelles by a higher fluidity of acyl chains and higher hydration/H-bonding of the interfacial carbonyl region. The presence of bile salts allowed observing changes in the IR spectrum of DPPC upon addition of PLRP2 (higher rigidity of acyl chains, dehydration of the interfacial carbonyl region), while no change was observed with Triton X-100. The differences between these surfactants and their impact on substrate recognition by PLRP2 are discussed, as well as the mechanism by which high and low molecular weight PLRP2-DPPC-bile salt complexes may be involved in the overall process of DPPC hydrolysis. Copyright © 2017 Elsevier B.V. All rights reserved.

77 FR 14417 - Notice of Temporary Closure on Public Lands in Gooding and Elmore Counties, ID

Federal Register 2010, 2011, 2012, 2013, 2014

2012-03-09

... affected public lands is: Boise Meridian, Idaho T. 3 S., R. 10 E., Sec. 35. T. 4 S., R. 10 E., Secs. 1 and 2; Secs. 12 to 15, inclusive; Sec. 17; Secs. 20 to 28, inclusive; Secs. 33 to 35, inclusive. T. 5 S., R. 10 E., Secs. 3 and 4. T. 3 S., R. 11 E., Secs. 31 and 33. T. 4 S., R. 11 E., Secs. 3 to 15...

Pancreatic SEC23B deficiency is sufficient to explain the perinatal lethality of germline SEC23B deficiency in mice

PubMed Central

Khoriaty, Rami; Everett, Lesley; Chase, Jennifer; Zhu, Guojing; Hoenerhoff, Mark; McKnight, Brooke; Vasievich, Matthew P.; Zhang, Bin; Tomberg, Kärt; Williams, John; Maillard, Ivan; Ginsburg, David

2016-01-01

In humans, loss of function mutations in SEC23B result in Congenital Dyserythropoietic Anemia type II (CDAII), a disease limited to defective erythroid development. Patients with two nonsense SEC23B mutations have not been reported, suggesting that complete SEC23B deficiency might be lethal. We previously reported that SEC23B-deficient mice die perinatally, exhibiting massive pancreatic degeneration and that mice with hematopoietic SEC23B deficiency do not exhibit CDAII. We now show that SEC23B deficiency restricted to the pancreas is sufficient to explain the lethality observed in mice with global SEC23B-deficiency. Immunohistochemical stains demonstrate an acinar cell defect but normal islet cells. Mammalian genomes contain two Sec23 paralogs, Sec23A and Sec23B. The encoded proteins share ~85% amino acid sequence identity. We generate mice with pancreatic SEC23A deficiency and demonstrate that these mice survive normally, exhibiting normal pancreatic weights and histology. Taken together, these data demonstrate that SEC23B but not SEC23A is essential for murine pancreatic development. We also demonstrate that two BAC transgenes spanning Sec23b rescue the lethality of mice homozygous for a Sec23b gene trap allele, excluding a passenger gene mutation as the cause of the pancreatic lethality, and indicating that the regulatory elements critical for Sec23b pancreatic function reside within the BAC transgenes. PMID:27297878

Legionella oakridgensis ATCC 33761 genome sequence and phenotypic characterization reveals its replication capacity in amoebae.

PubMed

Brzuszkiewicz, Elzbieta; Schulz, Tino; Rydzewski, Kerstin; Daniel, Rolf; Gillmaier, Nadine; Dittmann, Christine; Holland, Gudrun; Schunder, Eva; Lautner, Monika; Eisenreich, Wolfgang; Lück, Christian; Heuner, Klaus

2013-12-01

Legionella oakridgensis is able to cause Legionnaires' disease, but is less virulent compared to L. pneumophila strains and very rarely associated with human disease. L. oakridgensis is the only species of the family legionellae which is able to grow on media without additional cysteine. In contrast to earlier publications, we found that L. oakridgensis is able to multiply in amoebae. We sequenced the genome of L. oakridgensis type strain OR-10 (ATCC 33761). The genome is smaller than the other yet sequenced Legionella genomes and has a higher G+C-content of 40.9%. L. oakridgensis lacks a flagellum and it also lacks all genes of the flagellar regulon except of the alternative sigma-28 factor FliA and the anti-sigma-28 factor FlgM. Genes encoding structural components of type I, type II, type IV Lvh and type IV Dot/Icm, Sec- and Tat-secretion systems could be identified. Only a limited set of Dot/Icm effector proteins have been recognized within the genome sequence of L. oakridgensis. Like in L. pneumophila strains, various proteins with eukaryotic motifs and eukaryote-like proteins were detected. We could demonstrate that the Dot/Icm system is essential for intracellular replication of L. oakridgensis. Furthermore, we identified new putative virulence factors of Legionella. Copyright © 2013 Elsevier GmbH. All rights reserved.

An Axiology of Information Security for Futuristic Neuroprostheses: Upholding Human Values in the Context of Technological Posthumanization

PubMed Central

Gladden, Matthew E.

2017-01-01

Previous works exploring the challenges of ensuring information security for neuroprosthetic devices and their users have typically built on the traditional InfoSec concept of the “CIA Triad” of confidentiality, integrity, and availability. However, we argue that the CIA Triad provides an increasingly inadequate foundation for envisioning information security for neuroprostheses, insofar as it presumes that (1) any computational systems to be secured are merely instruments for expressing their human users' agency, and (2) computing devices are conceptually and practically separable from their users. Drawing on contemporary philosophy of technology and philosophical and critical posthumanist analysis, we contend that futuristic neuroprostheses could conceivably violate these basic InfoSec presumptions, insofar as (1) they may alter or supplant their users' biological agency rather than simply supporting it, and (2) they may structurally and functionally fuse with their users to create qualitatively novel “posthumanized” human-machine systems that cannot be secured as though they were conventional computing devices. Simultaneously, it is noted that many of the goals that have been proposed for future neuroprostheses by InfoSec researchers (e.g., relating to aesthetics, human dignity, authenticity, free will, and cultural sensitivity) fall outside the scope of InfoSec as it has historically been understood and touch on a wide range of ethical, aesthetic, physical, metaphysical, psychological, economic, and social values. We suggest that the field of axiology can provide useful frameworks for more effectively identifying, analyzing, and prioritizing such diverse types of values and goods that can (and should) be pursued through InfoSec practices for futuristic neuroprostheses. PMID:29163010

An Axiology of Information Security for Futuristic Neuroprostheses: Upholding Human Values in the Context of Technological Posthumanization.

PubMed

Gladden, Matthew E

2017-01-01

Previous works exploring the challenges of ensuring information security for neuroprosthetic devices and their users have typically built on the traditional InfoSec concept of the "CIA Triad" of confidentiality, integrity, and availability. However, we argue that the CIA Triad provides an increasingly inadequate foundation for envisioning information security for neuroprostheses, insofar as it presumes that (1) any computational systems to be secured are merely instruments for expressing their human users' agency, and (2) computing devices are conceptually and practically separable from their users. Drawing on contemporary philosophy of technology and philosophical and critical posthumanist analysis, we contend that futuristic neuroprostheses could conceivably violate these basic InfoSec presumptions, insofar as (1) they may alter or supplant their users' biological agency rather than simply supporting it, and (2) they may structurally and functionally fuse with their users to create qualitatively novel "posthumanized" human-machine systems that cannot be secured as though they were conventional computing devices. Simultaneously, it is noted that many of the goals that have been proposed for future neuroprostheses by InfoSec researchers (e.g., relating to aesthetics, human dignity, authenticity, free will, and cultural sensitivity) fall outside the scope of InfoSec as it has historically been understood and touch on a wide range of ethical, aesthetic, physical, metaphysical, psychological, economic, and social values. We suggest that the field of axiology can provide useful frameworks for more effectively identifying, analyzing, and prioritizing such diverse types of values and goods that can (and should) be pursued through InfoSec practices for futuristic neuroprostheses.

A Design and Development of Multi-Purpose CCD Camera System with Thermoelectric Cooling: Software

NASA Astrophysics Data System (ADS)

Oh, S. H.; Kang, Y. W.; Byun, Y. I.

2007-12-01

We present a software which we developed for the multi-purpose CCD camera. This software can be used on the all 3 types of CCD - KAF-0401E (768×512), KAF-1602E (15367times;1024), KAF-3200E (2184×1472) made in KODAK Co.. For the efficient CCD camera control, the software is operated with two independent processes of the CCD control program and the temperature/shutter operation program. This software is designed to fully automatic operation as well as manually operation under LINUX system, and is controled by LINUX user signal procedure. We plan to use this software for all sky survey system and also night sky monitoring and sky observation. As our results, the read-out time of each CCD are about 15sec, 64sec, 134sec for KAF-0401E, KAF-1602E, KAF-3200E., because these time are limited by the data transmission speed of parallel port. For larger format CCD, the data transmission is required more high speed. we are considering this control software to one using USB port for high speed data transmission.

78 FR 58554 - Notice of Application for Withdrawal Extension, and Notification of a Public Meeting, Desert...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-09-24

...., partially surveyed, Sec. 7, NW\\1/4\\ and S\\1/2\\. T. 16 S., R. 58 E., unsurveyed, Secs. 11 thru 14, inclusive; Secs. 23 thru 26, inclusive; Secs. 35 and 36. T. 15 S., R. 59 E., unsurveyed, Secs. 2 thru 11, inclusive; Secs. 14 thru 23, inclusive; Secs. 26 thru 35, inclusive. T. 16 S., R. 59 E., unsurveyed, Secs. 2...

Two distinct secretion systems facilitate tissue invasion by the rice blast fungus Magnaporthe oryzae

PubMed Central

Giraldo, Martha C.; Dagdas, Yasin F.; Gupta, Yogesh K.; Mentlak, Thomas A.; Yi, Mihwa; Martinez-Rocha, Ana Lilia; Saitoh, Hiromasa; Terauchi, Ryohei; Talbot, Nicholas J.; Valent, Barbara

2013-01-01

To cause plant diseases, pathogenic micro-organisms secrete effector proteins into host tissue to suppress immunity and support pathogen growth. Bacterial pathogens have evolved several distinct secretion systems to target effector proteins, but whether fungi, which cause the major diseases of most crop species, also require different secretory mechanisms is not known. Here we report that the rice blast fungus Magnaporthe oryzae possesses two distinct secretion systems to target effectors during plant infection. Cytoplasmic effectors, which are delivered into host cells, preferentially accumulate in the biotrophic interfacial complex, a novel plant membrane-rich structure associated with invasive hyphae. We show that the biotrophic interfacial complex is associated with a novel form of secretion involving exocyst components and the Sso1 t-SNARE. By contrast, effectors that are secreted from invasive hyphae into the extracellular compartment follow the conventional secretory pathway. We conclude that the blast fungus has evolved distinct secretion systems to facilitate tissue invasion. PMID:23774898

Roles of the conserved cytoplasmic region and non-conserved carboxy-terminal region of SecE in Escherichia coli protein translocase.

PubMed

Kontinen, V P; Yamanaka, M; Nishiyama, K; Tokuda, H

1996-06-01

SecE, an essential membrane component of the Escherichia coli protein translocase, consists of 127 amino acid residues. Only a part of the second putative cytoplasmic region comprising some 13 residues is essential for the SecE function as long as the proper topological arrangement is retained. The Trp84 and Pro85 residues of this region are conserved in all eubacterial SecE homologues. The conservation of positively charged residues corresponding to Arg80 and Lys81 is also substantial. We deleted or replaced these residues to assess their roles in the SecE function. Deletion of the Arg80-Lys81 dipeptide did not abolish the SecE function whereas that of Trp84 or Pro85 caused a loss of the function. Strikingly, however, replacement of Pro85 with either Gly, Ser, or Ala, and that of Trp84 with Lys did not abolish the SecE function. These results indicate that the strong conservation of these residues does not reflect their obligatory requirement for the SecE function. A chimeric SecE possessing the cytoplasmic region of the E. coli SecE and the following region of the Bacillus subtilis SecE was able to form the translocation machinery together with SecA, SecY, and SecG. Although a Leu to Arg mutation at position 108 has been thought to cause a loss of signal recognition fidelity and thereby suppress a signal sequence defect, the same mutation at position 111 caused a complete loss of the function. The levels of SecY and SecG in the secEcsE501 mutant, which expresses SecE at a decreased level and is sensitive to low temperature, increased upon the expression of functional SecE derivatives, irrespective of the site of mutation, suggesting that the levels of SecY and SecG are co-operatively determined by the level of functional, but not non-functional, SecE. Based on these results, the SecE function in the translocase is discussed.

Genotype and enterotoxigenicity of Staphylococcus epidermidis isolate from ready to eat meat products

PubMed Central

Podkowik, Magdalena; Seo, Keun Seok; Schubert, Justyna; Tolo, Isaiah; Robinson, D. Ashley; Bania, Jacek; Bystroń, Jarosław

2016-01-01

We have previously shown that potentially pathogenic isolates of Staphylococcus epidermidis occur at high incidence in ready-to-eat food. Now, within 164 samples of ready-to-eat meat products we identified 32 S. epidermidis isolates. In 8 isolates we detected the genes encoding for staphylococcal enterotoxins, but in 7 S. epidermidis isolates these genes were not stable over passages. One isolate designated 4S was shown to stably harbour sec and sel genes.In the genome sequence of S. epidermidis 4S we identified 21,426-bp region flanked by direct-repeats, encompassing sec and sel genes, corresponding to the previously described composite staphylococcal pathogenicity island (SePI) in S. epidermidis FRI909. Alignment of S. epidermidis 4S and S. epidermidis FRI909 SePIs revealed 6 nucleotide mismatches located in 5 of the total of 29 ORFs. Genomic location of S. epidermidis 4S SePI was the same as in FRI909. S. epidermidis 4S is a single locus variant of ST561, being genetically different from FRI909. SECepi was secreted by S. epidermidis 4S to BHI broth ranging from 14 to almost 36 μg/mL, to milk ranging from 6-9 ng/mL, to beef meat juice from 2-3 μg/mL and to pork meat juice from 1-2 μg/mL after 24 and 48 hours of cultivation, respectively. We provide the first evidence that S. epidermidis occurring in food bears an element encoding an orthologue to S. aureus SEC, and that SECepi can be produced in microbial broth, milk and meat juices. Regarding that only enterotoxins produced by S. aureus are officially tracked in food in EU, the ability to produce enterotoxin by S. epidermidis pose real risk for food safety. PMID:27105039

Genotype and enterotoxigenicity of Staphylococcus epidermidis isolate from ready to eat meat products.

PubMed

Podkowik, Magdalena; Seo, Keun Seok; Schubert, Justyna; Tolo, Isaiah; Robinson, D Ashley; Bania, Jacek; Bystroń, Jarosław

2016-07-16

We have previously shown that potentially pathogenic isolates of Staphylococcus epidermidis occur at high incidence in ready-to-eat food. Now, within 164 samples of ready-to-eat meat products we identified 32 S. epidermidis isolates. In 8 isolates we detected the genes encoding for staphylococcal enterotoxins, but in 7 S. epidermidis isolates these genes were not stable over passages. One isolate designated 4S was shown to stably harbour sec and sel genes. In the genome sequence of S. epidermidis 4S we identified 21,426-bp region flanked by direct-repeats, encompassing sec and sel genes, corresponding to the previously described composite staphylococcal pathogenicity island (SePI) in S. epidermidis FRI909. Alignment of S. epidermidis 4S and S. epidermidis FRI909 SePIs revealed 6 nucleotide mismatches located in 5 of the total of 29 ORFs. Genomic location of S. epidermidis 4S SePI was the same as in FRI909. S. epidermidis 4S is a single locus variant of ST561, being genetically different from FRI909. SECepi was secreted by S. epidermidis 4S to BHI broth ranging from 14 to almost 36μg/mL, to milk ranging from 6 to 9ng/mL, to beef meat juice from 2 to 3μg/mL and to pork meat juice from 1 to 2μg/mL after 24 and 48h of cultivation, respectively. We provide the first evidence that S. epidermidis occurring in food bears an element encoding an orthologue to Staphylococcus aureus SEC, and that SECepi can be produced in microbial broth, milk and meat juices. Regarding that only enterotoxins produced by S. aureus are officially tracked in food in EU, the ability to produce enterotoxin by S. epidermidis pose real risk for food safety. Copyright © 2016 Elsevier B.V. All rights reserved.

Determination of the molecular weight of low-molecular-weight heparins by using high-pressure size exclusion chromatography on line with a triple detector array and conventional methods.

PubMed

Bisio, Antonella; Mantegazza, Alessandra; Vecchietti, Davide; Bensi, Donata; Coppa, Alessia; Torri, Giangiacomo; Bertini, Sabrina

2015-03-19

The evaluation of weight average molecular weight (Mw) and molecular weight distribution represents one of the most controversial aspects concerning the characterization of low molecular weight heparins (LMWHs). As the most commonly used method for the measurement of such parameters is high performance size exclusion chromatography (HP-SEC), the soundness of results mainly depends on the appropriate calibration of the chromatographic columns used. With the aim of meeting the requirement of proper Mw standards for LMWHs, in the present work the determination of molecular weight parameters (Mw and Mn) by HP-SEC combined with a triple detector array (TDA) was performed. The HP-SEC/TDA technique permits the evaluation of polymeric samples by exploiting the combined and simultaneous action of three on-line detectors: light scattering detectors (LALLS/RALLS); refractometer and viscometer. Three commercial LMWH samples, enoxaparin, tinzaparin and dalteparin, a γ-ray depolymerized heparin (γ-Hep) and its chromatographic fractions, and a synthetic pentasaccharide were analysed by HP-SEC/TDA. The same samples were analysed also with a conventional HP-SEC method employing refractive index (RI) and UV detectors and two different chromatographic column set, silica gel and polymeric gel columns. In both chromatographic systems, two different calibration curves were built up by using (i) γ-Hep chromatographic fractions and the corresponding Mw parameters obtained via HP-SEC/TDA; (ii) the whole γ-Hep preparation with broad Mw dispersion and the corresponding cumulative distribution function calculated via HP-SEC/TDA. In addition, also a chromatographic column calibration according to European Pharmacopoeia indication was built up. By comparing all the obtained results, some important differences among Mw and size distribution values of the three LMWHs were found with the five different calibration methods and with HP-SEC/TDA method. In particular, the detection of the lower molecular weight components turned out to be the most critical aspect. Whereas HP-SEC/TDA may underestimate species under 2 KDa when present in low concentration, other methods appeared to emphasize their content.

The complete general secretory pathway in gram-negative bacteria.

PubMed Central

Pugsley, A P

1993-01-01

The unifying feature of all proteins that are transported out of the cytoplasm of gram-negative bacteria by the general secretory pathway (GSP) is the presence of a long stretch of predominantly hydrophobic amino acids, the signal sequence. The interaction between signal sequence-bearing proteins and the cytoplasmic membrane may be a spontaneous event driven by the electrochemical energy potential across the cytoplasmic membrane, leading to membrane integration. The translocation of large, hydrophilic polypeptide segments to the periplasmic side of this membrane almost always requires at least six different proteins encoded by the sec genes and is dependent on both ATP hydrolysis and the electrochemical energy potential. Signal peptidases process precursors with a single, amino-terminal signal sequence, allowing them to be released into the periplasm, where they may remain or whence they may be inserted into the outer membrane. Selected proteins may also be transported across this membrane for assembly into cell surface appendages or for release into the extracellular medium. Many bacteria secrete a variety of structurally different proteins by a common pathway, referred to here as the main terminal branch of the GSP. This recently discovered branch pathway comprises at least 14 gene products. Other, simpler terminal branches of the GSP are also used by gram-negative bacteria to secrete a more limited range of extracellular proteins. PMID:8096622

Export requirements of pneumolysin in Streptococcus pneumoniae.

PubMed

Price, Katherine E; Greene, Neil G; Camilli, Andrew

2012-07-01

Streptococcus pneumoniae is a major causative agent of otitis media, pneumonia, bacteremia, and meningitis. Pneumolysin (Ply), a member of the cholesterol-dependent cytolysins (CDCs), is produced by virtually all clinical isolates of S. pneumoniae, and ply mutant strains are severely attenuated in mouse models of colonization and infection. In contrast to all other known members of the CDC family, Ply lacks a signal peptide for export outside the cell. Instead, Ply has been hypothesized to be released upon autolysis or, alternatively, via a nonautolytic mechanism that remains undefined. We show that an exogenously added signal sequence is not sufficient for Sec-dependent Ply secretion in S. pneumoniae but is sufficient in the surrogate host Bacillus subtilis. Previously, we showed that Ply is localized primarily to the cell wall compartment in the absence of detectable cell lysis. Here we show that Ply released by autolysis cannot reassociate with intact cells, suggesting that there is a Ply export mechanism that is coupled to cell wall localization of the protein. This putative export mechanism is capable of secreting a related CDC without its signal sequence. We show that B. subtilis can export Ply, suggesting that the export pathway is conserved. Finally, through truncation and domain swapping analyses, we show that export is dependent on domain 2 of Ply.

Export Requirements of Pneumolysin in Streptococcus pneumoniae

PubMed Central

Price, Katherine E.; Greene, Neil G.

2012-01-01

Streptococcus pneumoniae is a major causative agent of otitis media, pneumonia, bacteremia, and meningitis. Pneumolysin (Ply), a member of the cholesterol-dependent cytolysins (CDCs), is produced by virtually all clinical isolates of S. pneumoniae, and ply mutant strains are severely attenuated in mouse models of colonization and infection. In contrast to all other known members of the CDC family, Ply lacks a signal peptide for export outside the cell. Instead, Ply has been hypothesized to be released upon autolysis or, alternatively, via a nonautolytic mechanism that remains undefined. We show that an exogenously added signal sequence is not sufficient for Sec-dependent Ply secretion in S. pneumoniae but is sufficient in the surrogate host Bacillus subtilis. Previously, we showed that Ply is localized primarily to the cell wall compartment in the absence of detectable cell lysis. Here we show that Ply released by autolysis cannot reassociate with intact cells, suggesting that there is a Ply export mechanism that is coupled to cell wall localization of the protein. This putative export mechanism is capable of secreting a related CDC without its signal sequence. We show that B. subtilis can export Ply, suggesting that the export pathway is conserved. Finally, through truncation and domain swapping analyses, we show that export is dependent on domain 2 of Ply. PMID:22563048

Online Detection of Functional Groups in SEC via Quantum Cascade Laser IR Spectroscopy.

PubMed

Morlock, Sascha; Kübel, Jennifer M; Beskers, Timo F; Lendl, Bernhard; Wilhelm, Manfred

2018-01-01

The development of coupled techniques based on chemically sensitive detectors, such as FTIR or NMR spectrometers, for size exclusion chromatography (SEC) provides sophisticated methods for determining the molecular-weight-dependent chemical composition in polymers. However, the detection of rare functionalities such as end groups or branching points presents a challenge, especially for online coupled SEC detection, which is based on low-concentration chromatography. To address this issue, for the first time, an external cavity quantum cascade laser (EC-QCL) infrared spectrometer is coupled to an SEC. The system is evaluated using polystyrene/poly(methyl methacrylate) (PS/PMMA) blends. The current limit of detection for the carbonyl (PMMA) stretch vibration at 1730 cm -1 with this technique is 3.5 µg PMMA on a semipreparative column (typical load of 2.5 mg polymer in total). That equals 0.15 mol% of PMMA in the PS/PMMA blend and corresponds to one carbonyl group per 70 kg mol -1 polymer. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Roles of silkworm endoplasmic reticulum chaperones in the secretion of recombinant proteins expressed by baculovirus system.

PubMed

Imai, Saki; Kusakabe, Takahiro; Xu, Jian; Li, Zhiqing; Shirai, Shintaro; Mon, Hiroaki; Morokuma, Daisuke; Lee, Jae Man

2015-11-01

Baculovirus expression vector system (BEVS) is widely used for production of recombinant eukaryotic proteins in insect larvae or cultured cells. BEVS has advantages over bacterial expression system in producing post-translationally modified secreted proteins. However, for some unknown reason, it is very difficult for insects to secrete sufficiently for certain proteins of interest. To understand the reasons why insect cells fail to secrete some kinds of recombinant proteins, we here employed three mammalian proteins as targets, EPO, HGF, and Wnt3A, with different secretion levels in BEVS and investigated their mRNA transcriptions from the viral genome, subcellular localizations, and interactions with silkworm ER chaperones. Moreover, we observed that no significantly influence on the secretion amounts of all three proteins when depleting or overexpressing most endogenous ER chaperone genes in cultured silkworm cells. However, among all detected ER chaperones, the depletion of BiP severely decreased the recombinant protein secretion in BEVS, indicating the possible central role of Bip in silkworm secretion pathway.

77 FR 36611 - Core Principles and Other Requirements for Designated Contract Markets

Federal Register 2010, 2011, 2012, 2013, 2014

2012-06-19

.... 38.154--Regulatory Services Provided by a Third Party vi. Sec. 38.155--Compliance Staff and Resources... vi. Sec. 38.256--Trade Reconstruction vii. Sec. 38.257--Regulatory Service Provider viii. Sec. 38.258... Integrity iii. Sec. 38.603--Protection of Customer Funds iv. Sec. 38.604--Financial Surveillance v. Sec. 38...

System Specificity of the TpsB Transporters of Coexpressed Two-Partner Secretion Systems of Neisseria meningitidis

PubMed Central

ur Rahman, Sadeeq

2013-01-01

The two-partner secretion (TPS) systems of Gram-negative bacteria consist of a large secreted exoprotein (TpsA) and a transporter protein (TpsB) located in the outer membrane. TpsA targets TpsB for transport across the membrane via its ∼30-kDa TPS domain located at its N terminus, and this domain is also the minimal secretory unit. Neisseria meningitidis genomes encode up to five TpsAs and two TpsBs. Sequence alignments of TPS domains suggested that these are organized into three systems, while there are two TpsBs, which raised questions on their system specificity. We show here that the TpsB2 transporter of Neisseria meningitidis is able to secrete all types of TPS domains encoded in N. meningitidis and the related species Neisseria lactamica but not domains of Haemophilus influenzae and Pseudomonas aeruginosa. In contrast, the TpsB1 transporter seemed to be specific for its cognate N. meningitidis system and did not secrete the TPS domains of other meningococcal systems. However, TpsB1 did secrete the TPS2b domain of N. lactamica, which is related to the meningococcal TPS2 domains. Apparently, the secretion depends on specific sequences within the TPS domain rather than the overall TPS domain structure. PMID:23222722

Response of a small-turboshaft-engine compression system to inlet temperature distortion

NASA Technical Reports Server (NTRS)

Biesiadny, T. J.; Klann, G. A.; Little, J. K.

1984-01-01

An experimental investigation was conducted into the response of a small-turboshaft-engine compression system to steady-state and transient inlet temperature distortions. Transient temperature ramps range from less than 100 K/sec to above 610 K/sec and generated instantaneous temperatures to 420 K above ambient. Steady-state temperature distortion levels were limited by the engine hardware temperature list. Simple analysis of the steady-state distortion data indicated that a particle separator at the engine inlet permitted higher levels of temperature distortion before onset of compressor surge than would be expected without the separator.

Dabigatran exhibits low intensity of left atrial spontaneous echo contrast in patients with nonvalvular atrial fibrillation as compared with warfarin.

PubMed

Watanabe, Tetsuya; Shinoda, Yukinori; Ikeoka, Kuniyasu; Inui, Hirooki; Fukuoka, Hidetada; Sunaga, Akihiro; Kanda, Takashi; Uematsu, Masaaki; Hoshida, Shiro

2017-03-01

The presence of spontaneous echo contrast (SEC) in the left atrium has been reported to be an independent predictor of thromboembolic risk in patients with atrial fibrillation (AF). Dabigatran was associated with lower rates of stroke and systemic embolism as compared with warfarin when administered at a higher dose. Between July 2011 and October 2015, nonvalvular AF patients treated with warfarin or dabigatran who had transesophageal echocardiography prior to ablation therapy for AF were enrolled. The intensity of SEC was classified into four grades, from 0 to 3. Univariate and multivariate analysis was performed to analyze factors associated with SEC. Sixty-five patients were on dabigatran and 65 were on warfarin, with the prothrombin time in therapeutic range. There were no significant differences in the age, CHADS2 score, left atrial dimension, and left atrial appendage flow between the two groups. However, there were more grade 2 or higher patients with left atrial SEC in the warfarin group (n = 20) than in the dabigatran group (n = 2) (p < 0.001). When multivariate regression analysis was performed, grade 2 or higher left atrial SEC was independently associated with no dabigatran usage in addition to high brain natriuretic peptide level and high incidence of diabetes mellitus or persistent AF. Thus, dabigatran exhibited low intensity of left atrial SEC in nonvalvular AF patients as compared with warfarin.

GOLGI TRANSPORT 1B Regulates Protein Export from the Endoplasmic Reticulum in Rice Endosperm Cells[OPEN

PubMed Central

Liu, Feng; Wang, Yunlong; Liu, Xi; Wang, Di; Zhu, Xiaopin; Jing, Ruonan; Wu, Mingming; Hao, Yuanyuan; Jiang, Ling; Wang, Chunming

2016-01-01

Coat protein complex II (COPII) mediates the first step of anterograde transport of newly synthesized proteins from the endoplasmic reticulum (ER) to other endomembrane compartments in eukaryotes. A group of evolutionarily conserved proteins (Sar1, Sec23, Sec24, Sec13, and Sec31) constitutes the basic COPII coat machinery; however, the details of how the COPII coat assembly is regulated remain unclear. Here, we report a protein transport mutant of rice (Oryza sativa), named glutelin precursor accumulation4 (gpa4), which accumulates 57-kD glutelin precursors and forms two types of ER-derived abnormal structures. GPA4 encodes the evolutionarily conserved membrane protein GOT1B (also known as GLUP2), homologous to the Saccharomyces cerevisiae GOT1p. The rice GOT1B protein colocalizes with Arabidopsis thaliana Sar1b at Golgi-associated ER exit sites (ERESs) when they are coexpressed in Nicotiana benthamiana. Moreover, GOT1B physically interacts with rice Sec23, and both proteins are present in the same complex(es) with rice Sar1b. The distribution of rice Sar1 in the endomembrane system, its association with rice Sec23c, and the ERES organization pattern are significantly altered in the gpa4 mutant. Taken together, our results suggest that GOT1B plays an important role in mediating COPII vesicle formation at ERESs, thus facilitating anterograde transport of secretory proteins in plant cells. PMID:27803308

Considerations of digital phase modulation for narrowband satellite mobile communication

NASA Technical Reports Server (NTRS)

Grythe, Knut

1990-01-01

The Inmarsat-M system for mobile satellite communication is specified as a frequency division multiple access (FDMA) system, applying Offset Quadrature Phase Shift Keying (QPSK) for transmitting 8 kbit/sec in 10 kHz user channel bandwidth. We consider Digital Phase Modulation (DPM) as an alternative modulation format for INMARSAT-M. DPM is similar to Continuous Phase Modulation (CPM) except that DPM has a finite memory in the premodular filter with a continuous varying modulation index. It is shown that DPM with 64 states in the VA obtains a lower bit error rate (BER). Results for a 5 kHz system, with the same 8 kbit/sec transmitted bitstream, is also presented.

DBSecSys 2.0: a database of Burkholderia mallei and Burkholderia pseudomallei secretion systems.

PubMed

Memišević, Vesna; Kumar, Kamal; Zavaljevski, Nela; DeShazer, David; Wallqvist, Anders; Reifman, Jaques

2016-09-20

Burkholderia mallei and B. pseudomallei are the causative agents of glanders and melioidosis, respectively, diseases with high morbidity and mortality rates. B. mallei and B. pseudomallei are closely related genetically; B. mallei evolved from an ancestral strain of B. pseudomallei by genome reduction and adaptation to an obligate intracellular lifestyle. Although these two bacteria cause different diseases, they share multiple virulence factors, including bacterial secretion systems, which represent key components of bacterial pathogenicity. Despite recent progress, the secretion system proteins for B. mallei and B. pseudomallei, their pathogenic mechanisms of action, and host factors are not well characterized. We previously developed a manually curated database, DBSecSys, of bacterial secretion system proteins for B. mallei. Here, we report an expansion of the database with corresponding information about B. pseudomallei. DBSecSys 2.0 contains comprehensive literature-based and computationally derived information about B. mallei ATCC 23344 and literature-based and computationally derived information about B. pseudomallei K96243. The database contains updated information for 163 B. mallei proteins from the previous database and 61 additional B. mallei proteins, and new information for 281 B. pseudomallei proteins associated with 5 secretion systems, their 1,633 human- and murine-interacting targets, and 2,400 host-B. mallei interactions and 2,286 host-B. pseudomallei interactions. The database also includes information about 13 pathogenic mechanisms of action for B. mallei and B. pseudomallei secretion system proteins inferred from the available literature or computationally. Additionally, DBSecSys 2.0 provides details about 82 virulence attenuation experiments for 52 B. mallei secretion system proteins and 98 virulence attenuation experiments for 61 B. pseudomallei secretion system proteins. We updated the Web interface and data access layer to speed-up users' search of detailed information for orthologous proteins related to secretion systems of the two pathogens. The updates of DBSecSys 2.0 provide unique capabilities to access comprehensive information about secretion systems of B. mallei and B. pseudomallei. They enable studies and comparisons of corresponding proteins of these two closely related pathogens and their host-interacting partners. The database is available at http://dbsecsys.bhsai.org .

Agonal sequences in four filmed hangings: analysis of respiratory and movement responses to asphyxia by hanging.

PubMed

Sauvageau, Anny

2009-01-01

The human pathophysiology of asphyxia by hanging is still poorly understood, despite great advances in forensic science. In that context, filmed hangings may hold the key to answer questions regarding the sequence of events leading to death in human asphyxia. Four filmed hangings were analyzed. Rapid loss of consciousness was observed between 13 sec and 18 sec after onset of hanging, closely followed by convulsions (at 14-19 sec). A complex pattern of decerebration rigidity (19-21 sec in most cases), followed by a quick phase of decortication rigidity (1 min 00 sec-1 min 08 sec in most cases), an extended phase of decortication rigidity (1 min 04 sec-1 min 32 sec) and loss of muscle tone (1 min 38 sec-2 min 47 sec) was revealed. Very deep respiratory attempts started between 20 and 22 sec, the last respiratory attempt being detected between 2 min 00 sec and 2 min 04 sec. Despite differences in the types of hanging, this unique study reveals similarities that are further discussed.

Bleed cycle propellant pumping in a gas-core nuclear rocket engine system

NASA Technical Reports Server (NTRS)

Kascak, A. F.; Easley, A. J.

1972-01-01

The performance of ideal and real staged primary propellant pumps and bleed-powered turbines was calculated for gas-core nuclear rocket engines over a range of operating pressures from 500 to 5000 atm. This study showed that for a required engine operating pressure of 1000 atm the pump work was about 0.8 hp/(lb/sec), the specific impulse penalty resulting from the turbine propellant bleed flow as about 10 percent; and the heat required to preheat the propellant was about 7.8 MN/(lb/sec). For a specific impulse above 2400 sec, there is an excess of energy available in the moderator due to the gamma and neutron heating that occurs there. Possible alternative pumping cycles are the Rankine or Brayton cycles.

The analysis results of EEWS(Earthquake Early Warning System) about Iksan(Ml4.3) and Ulsan(Ml5.0) earthquakes in Korea

NASA Astrophysics Data System (ADS)

Park, J. H.; Chi, H. C.; Lim, I. S.; Seong, Y. J.; Pak, J.

2016-12-01

EEW(Earthquake Early Warning) service to the public has been officially operated by KMA (Korea Meteorological Administration) from 2015 in Korea. For the KMA's official EEW service, KIGAM has adopted ElarmS from UC Berkeley BSL and modified local magnitude relation, 1-D travel time curves and association procedures with real time waveforms from about 160 seismic stations of KMA and KIGAM. We have checked the performance of EEWS(Earthquake Early Warning System) reviewing two moderate size earthquakes: one is Iksan Eq.(Ml4.3) inside of networks and the other is Ulsan Eq.(Ml5.0) happened at the southern east sea of Korea outside of networks. The first trigger time at NPR station of the Iksan Eq. took 2.3 sec and BUY and JEO2 stations were associated to produce the first event version in 10.07 sec from the origin time respectively. Because the epicentral distance of JEO2 station is about 30 km and the estimated travel time is 6.2 sec, the delay time including transmission and processing is estimated as 3.87 sec with assumption that P wave velocity is 5 km/sec and the focal depth is 8 km. The first magnitude was M4.9 which was a little bigger than Ml4.3 by KIGAM. After adding 3 more triggers of stations (CHO, KMSA, PORA), the estimated magnitude became to M4.6 and the final was settled down to M4.3 with 10 stations. In the case of Ulsan the first trigger time took 11.04 sec and the first alert time with 3 stations in 14.8 sec from the origin time (OT) respectively. The first magnitude was M5.2, however, the difference between the first EEW epicenter and the manual final result was about 63 km due to the poor azimuth coverage outside of seismic network. After 16.2 sec from OT the fourth station YSB was used to update the location near to the manual results within 6 km with magnitude 5.0 and location and magnitude were stable with more stations. Ulsan Eq. was the first case announced to the public by EEWS and the process and result were successful, however, we have to make more effort to reduce the delay times and improve tools for the communication with the public in Korea.

ARGOT: A System Overview,

DTIC Science & Technology

1982-04-01

the secret . In each, we shall consider * the plausible interpretations of the utterance "Do you know the secret ?" Setting 1: If S knows the secret and...believes that H doesn’t know the secret , then "Do you know the secret ?" is probably an offer to tell H the secret . Setting 2: If S doesn’t know the ... secret and believes that H does know the secret then "Do you know the

Identification of O-linked β-d-N-acetylglucosamine-Modified Proteins from Arabidopsis

PubMed Central

Xu, Shou-Ling; Chalkley, Robert J.; Wang, Zhi-Yong; Burlingame, Alma L.

2013-01-01

The posttranslational modification of proteins with O-linked β-d-N-acetylglucosamine (O-GlcNAc) on serine and threonine residues occurs in all animals and plants. This modification is dynamic and ubiquitous, and regulates many cellular processes, including transcription, signaling and cytokinesis and is associated with several diseases. Cycling of O-GlcNAc is tightly regulated by O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA). Plants have two OGTs, SPINDLY (SPY) and SECRET AGENT (SEC); disruption of both causes embryo lethality. Despite O-GlcNAc modification of proteins being discovered more than 20-years ago, identification and mapping of protein GlcNAcylation is still a challenging task. Here we describe the use of lectin affinity chromatography combined with electron transfer dissociation mass spectrometry to enrich and to detect O-GlcNAc modified peptides from Arabidopsis. PMID:22576084

Ubiquitin-dependent regulation of COPII coat size and function

PubMed Central

Jin, Lingyan; Pahuja, Kanika Bajaj; Wickliffe, Katherine E.; Gorur, Amita; Baumgärtel, Christine; Schekman, Randy; Rape, Michael

2012-01-01

Packaging of proteins from the ER into COPII-vesicles is essential for secretion. In cells, most COPII-vesicles are ~60-80nm in diameter, yet some must increase their size to accommodate 300-400nm procollagen fibers or chylomicrons. Impaired COPII function results in collagen deposition defects, cranio-lenticulo-sutural dysplasia, or chylomicron retention disease, but mechanisms to enlarge COPII-coats have remained elusive. Here, we have identified the ubiquitin ligase Cul3Klhl12 as a regulator of COPII coat formation. Cul3Klhl12 catalyzes the monoubiquitination of the COPII-component Sec31 and drives the assembly of large COPII coats. As a result, ubiquitination by Cul3Klhl12 is essential for collagen export, yet less important for the transport of small cargo. We conclude that monoubiquitination controls the size and function of a vesicle coat. PMID:22358839

[Clinical significance of analysis of immunoglobulin A levels in saliva].

PubMed

Bokor-Bratić, M

2000-01-01

SALIVA COLLECTION: Whole saliva is a product of secretion of 3 major glands (parotid, submandibular, sublingual) and many minor glands (labial, buccal, palatal). Unstimulated saliva is usually obtained as the patient spits out every 60 sec. or by forward bended head the patient allows saliva to drip off the lower lip into a cylinder. By collection of saliva in the tube the flow rate per unit time can be measured. When volume measurement is not required the saliva can be collected on cotton rolls, gauze or filter paper. For evaluating salivary gland function or when large volumes of saliva are required for analytic purposes, stimulated whole saliva is used. Method of collection is the same as for unstimulated saliva. The usual masticatory stimuli are paraffin wax or a washed rubber band. A standard gustatory stimulus is obtained by 2% citric acid applied directly to the tongue every 15 to 60 sec. Parotid saliva can be collected by aspiration from the duct opening with a micropipette. Parotid saliva is best collected with Lashley's vacuum chamber. Submandibular and sublingual saliva can be collected by cannulation of the duct with micropipette, but in practice this is both uncomfortable for the patients and technically difficult since the duct orifice is mobile and has a strong sphincter. Because of that, alginate and silicone impression material is used for retention of the collecting tube. As alternative and simple technique is to block off secretion from the parotid glands with absorbent swabs and collect mixed submandibular and sublingual saliva by pipette from the floor of the mouth. Saliva from labial and palatal glands can be collected by filter paper disc or disc of other synthetic materials. SALIVARY IMMUNOGLOBULIN A: The most significant characteristics of the salivary immunoglobulin system are quantitative domination of immunoglobulin A, local synthesis and specific structure. Immunofluorescence studies have shown that immunoglobulin A is produced by plasma cells locally in the salivary glands. There is still little convincing evidence for the origin of predominantly immunoglobulin A secreting plasma cells in salivary glands. DETECTION OF IMMUNOGLOBULIN A IN SALIVA: Radial immunodiffusion (RID) was the most applicable method for detecting salivary immunoglobulin A. However, there are more sensitive and automatic methods such as nephelometry and ELISA. A standard level of immunoglobulin in saliva is still in question since the concentration varies in relation to origin of saliva, method of collection and stimulation of secretion (Table 1). PERIODONTAL DISEASE: Studies of the salivary immunoglobulin A in patients with periodontal disease and healthy persons showed that there are differences which can be used in detection of high-risk groups and individuals. If the bacterial adherence to the mucosa is a prerequisite for bacterial evolution in subgingival or any other region of the oral cavity respectively introduction in periodontitis development, than it is to be presumed that the basic function of salivary immunoglobulin A is inhibition of bacterial adherence rather than antigens destruction. Several bacterial species frequently isolated from the oral cavity of patients with periodontitis have been identified as producers of IgA protease. These enzymes cleave serum IgA and secretory IgA equally well. Additionally, most of the IgA proteases studied have cleaved the A1 and A2 subclass. Several studies have demonstrated that cleavage of human IgA occurs in vivo, resulting in generation of intact Fab alpha and (Fc alpha)2 fragment. Moreover, when bacteria are exposed to Fab alpha fragments released from IgA after cleavage by IgA protease, their surface antigens are likely to be occupied by Fab alpha fragments. These Fab alpha fragments left on the bacterial surface may mediate adhesion. Together, these results indicate that IgA proteases, by promoting adherence, contribute the pathogenic potential of bacteria in the oral c

The New Kid on the Block: A Specialized Secretion System during Bacterial Sporulation.

PubMed

Morlot, Cécile; Rodrigues, Christopher D A

2018-02-02

The transport of proteins across the bacterial cell envelope is mediated by protein complexes called specialized secretion systems. These nanomachines exist in both Gram-positive and Gram-negative bacteria and have been categorized into different types based on their structural components and function. Interestingly, multiple studies suggest the existence of a protein complex in endospore-forming bacteria that appears to be a new type of specialized secretion system. This protein complex is called the SpoIIIA-SpoIIQ complex and is an exception to the categorical norm since it appears to be a hybrid composed of different parts from well-defined specialized secretion systems. Here we summarize and discuss the current understanding of this complex and its potential role as a specialized secretion system. Copyright © 2018 Elsevier Ltd. All rights reserved.

Diversity of secretion systems associated with virulence characteristics of the classical bordetellae.

PubMed

Park, Jihye; Zhang, Ying; Chen, Chun; Dudley, Edward G; Harvill, Eric T

2015-12-01

Secretion systems are key virulence factors, modulating interactions between pathogens and the host's immune response. Six potential secretion systems (types 1-6; T1SS-T6SS) have been discussed in classical bordetellae, respiratory commensals/pathogens of mammals. The prototypical Bordetella bronchiseptica strain RB50 genome seems to contain all six systems, whilst two human-restricted subspecies, Bordetella parapertussis and Bordetella pertussis, have lost different subsets of these. This implicates secretion systems in the divergent evolutionary histories that have led to their success in different niches. Based on our previous work demonstrating that changes in secretion systems are associated with virulence characteristics, we hypothesized there would be substantial divergence of the loci encoding each amongst sequenced strains. Here, we describe extensive differences in secretion system loci; 10 of the 11 sequenced strains had lost subsets of genes or one entire secretion system locus. These loci contained genes homologous to those present in the respective loci in distantly related organisms, as well as genes unique to bordetellae, suggesting novel and/or auxiliary functions. The high degree of conservation of the T3SS locus, a complex machine with interdependent parts that must be conserved, stands in dramatic contrast to repeated loss of T5aSS 'autotransporters', which function as an autonomous unit. This comparative analysis provided insights into critical aspects of each pathogen's adaptation to its different niche, and the relative contributions of recombination, mutation and horizontal gene transfer. In addition, the relative conservation of various secretion systems is an important consideration in the ongoing search for more highly conserved protective antigens for the next generation of pertussis vaccines.

Salmonella-secreted Virulence Factors

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

Heffron, Fred; Niemann, George; Yoon, Hyunjin

In this short review we discuss secreted virulence factors of Salmonella, which directly affect Salmonella interaction with its host. Salmonella secretes protein to subvert host defenses but also, as discussed, to reduce virulence thereby permitting the bacteria to persist longer and more successfully disperse. The type III secretion system (TTSS) is the best known and well studied of the mechanisms that enable secretion from the bacterial cytoplasm to the host cell cytoplasm. Other secretion systems include outer membrane vesicles, which are present in all Gram-negative bacteria examined to date, two-partner secretion, and type VI secretion will also be addressed. Excellentmore » reviews of Salmonella secreted effectors have focused on themes such as actin rearrangements, vesicular trafficking, ubiquitination, and the activities of the virulence factors themselves. This short review is based on S. Typhimurium infection of mice because it is a model of typhoid like disease in humans. We have organized effectors in terms of events that happen during the infection cycle and how secreted effectors may be involved.« less

Proteinaceous determinants of surface colonization in bacteria: bacterial adhesion and biofilm formation from a protein secretion perspective

PubMed Central

Chagnot, Caroline; Zorgani, Mohamed A.; Astruc, Thierry; Desvaux, Mickaël

2013-01-01

Bacterial colonization of biotic or abiotic surfaces results from two quite distinct physiological processes, namely bacterial adhesion and biofilm formation. Broadly speaking, a biofilm is defined as the sessile development of microbial cells. Biofilm formation arises following bacterial adhesion but not all single bacterial cells adhering reversibly or irreversibly engage inexorably into a sessile mode of growth. Among molecular determinants promoting bacterial colonization, surface proteins are the most functionally diverse active components. To be present on the bacterial cell surface, though, a protein must be secreted in the first place. Considering the close association of secreted proteins with their cognate secretion systems, the secretome (which refers both to the secretion systems and their protein substrates) is a key concept to apprehend the protein secretion and related physiological functions. The protein secretion systems are here considered in light of the differences in the cell-envelope architecture between diderm-LPS (archetypal Gram-negative), monoderm (archetypal Gram-positive) and diderm-mycolate (archetypal acid-fast) bacteria. Besides, their cognate secreted proteins engaged in the bacterial colonization process are regarded from single protein to supramolecular protein structure as well as the non-classical protein secretion. This state-of-the-art on the complement of the secretome (the secretion systems and their cognate effectors) involved in the surface colonization process in diderm-LPS and monoderm bacteria paves the way for future research directions in the field. PMID:24133488

In Candida albicans hyphae, Sec2p is physically associated with SEC2 mRNA on secretory vesicles.

PubMed

Caballero-Lima, David; Hautbergue, Guillaume M; Wilson, Stuart A; Sudbery, Peter E

2014-11-01

Candida albicans hyphae grow in a highly polarized fashion from their tips. This polarized growth requires the continuous delivery of secretory vesicles to the tip region. Vesicle delivery depends on Sec2p, the Guanine Exchange Factor (GEF) for the Rab GTPase Sec4p. GTP bound Sec4p is required for the transit of secretory vesicles from the trans-Golgi to sites of polarized growth. We previously showed that phosphorylation of Sec2p at residue S584 was necessary for Sec2p to support hyphal, but not yeast growth. Here we show that on secretory vesicles SEC2 mRNA is physically associated with Sec2p. Moreover, we show that the phosphorylation of S584 allows SEC2 mRNA to dissociate from Sec2p and we speculate that this is necessary for Sec2p function and/or translation. During hyphal extension, the growing tip may be separated from the nucleus by up to 15 μm. Transport of SEC2 mRNA on secretory vesicles to the tip localizes SEC2 translation to tip allowing a sufficient accumulation of this key protein at the site of polarized growth. © 2014 The Authors. Molecular Microbiology published by John Wiley & Sons Ltd.

Nonlethal sec71-1 and sec72-1 mutations eliminate proteins associated with the Sec63p-BiP complex from S. cerevisiae.

PubMed Central

Fang, H; Green, N

1994-01-01

The sec71-1 and sec72-1 mutations were identified by a genetic assay that monitored membrane protein integration into the endoplasmic reticulum (ER) membrane of the yeast Saccharomyces cerevisiae. The mutations inhibited integration of various chimeric membrane proteins and translocation of a subset of water soluble proteins. In this paper we show that SEC71 encodes the 31.5-kDa transmembrane glycoprotein (p31.5) and SEC72 encodes the 23-kDa protein (p23) of the Sec63p-BiP complex. SEC71 is therefore identical to SEC66 (HSS1), which was previously shown to encode p31.5. DNA sequence analyses reveal that sec71-1 cells contain a nonsense mutation that removes approximately two-thirds of the cytoplasmic C-terminal domain of p31.5. The sec72-1 mutation shifts the reading frame of the gene encoding p23. Unexpectedly, the sec71-1 mutant lacks p31.5 and p23. Neither mutation is lethal, although sec71-1 cells exhibit a growth defect at 37 degrees C. These results show that p31.5 and p23 are important for the trafficking of a subset of proteins to the ER membrane. Images PMID:7841522

Sec71 functions as a GEF for the small GTPase Arf1 to govern dendrite pruning of Drosophila sensory neurons.

PubMed

Wang, Yan; Zhang, Heng; Shi, Meng; Liou, Yih-Cherng; Lu, Lei; Yu, Fengwei

2017-05-15

Pruning, whereby neurons eliminate their excess neurites, is central for the maturation of the nervous system. In Drosophila , sensory neurons, ddaCs, selectively prune their larval dendrites without affecting their axons during metamorphosis. However, it is unknown whether the secretory pathway plays a role in dendrite pruning. Here, we show that the small GTPase Arf1, an important regulator of the secretory pathway, is specifically required for dendrite pruning of ddaC/D/E sensory neurons but dispensable for apoptosis of ddaF neurons. Analyses of the GTP- and GDP-locked forms of Arf1 indicate that the cycling of Arf1 between GDP-bound and GTP-bound forms is essential for dendrite pruning. We further identified Sec71 as a guanine nucleotide exchange factor for Arf1 that preferentially interacts with its GDP-bound form. Like Arf1, Sec71 is also important for dendrite pruning, but not for apoptosis, of sensory neurons. Arf1 and Sec71 are interdependent for their localizations on Golgi. Finally, we show that the Sec71/Arf1-mediated trafficking process is a prerequisite for Rab5-dependent endocytosis to facilitate endocytosis and degradation of the cell-adhesion molecule Neuroglian (Nrg). © 2017. Published by The Company of Biologists Ltd.

Flight assessment of a data-link-based navigation-guidance concept

NASA Technical Reports Server (NTRS)

Abbott, T. S.

1983-01-01

With the proposed introduction of a data-link provision into the Air-Traffic-control (ATC) system, the capability will exist to supplement the ground-air, voice (radio) link with digital, data-link information. Additionally, ATC computers could provide, via the data link guidance and navigation information to the pilot which could then be presented in much the same manner as conventional navigation information. The primary objective of this study was to assess the feasibility and acceptability of using 4-sec and 12-sec information updating to drive conventional cockpit-navigation-instrument formats for path-tracking guidance. A flight test, consisting of 19 tracking tasks, was conducted and, through the use of pilot questionnaires and performance data, the following results were obtained. From a performance standpoint, the 4-sec and 12-sec updating led to a slight degradation in path-tracking performance, relative to continuous updating. From the pilot's viewpoint, the 12-sec data interval was suitable for long path segments (greater than 2 min of flight time), but it was difficult to use on shorter segments because of higher work load and insufficient stabilization time. Overall, it was determined that the utilization of noncontinuous data for navigation was both feasible and acceptable for the prescribed task.

Outer satellite atmospheres: Their nature and planetary interactions

NASA Technical Reports Server (NTRS)

Smyth, W. H.; Combi, M. R.

1984-01-01

Significant insights regarding the nature and interactions of Io and the planetary magnetosphere were gained through modeling studies of the spatial morphology and brightness of the Io sodium cloud. East-west intensity asymmetries in Region A are consistent with an east-west electric field and the offset of the magnetic and planetary-spin axes. East-west orbital asymmetries and the absolute brightness of Region B suggest a low-velocity (3 km/sec) satellite source of 1 to 2 x 10(26) sodium atoms/sec. The time-varying spatial structure of the sodium directional features in near Region C provides direct evidence for a magnetospheric-wind-driven escape mechanism with a high-velocity (20 km/sec) source of 1 x 10(26) atoms/sec and a flux distribution enhanced at the equator relative to the poles. A model for the Io potassium cloud is presented and analysis of data suggests a low velocity source rate of 5 x 10(24) atoms/sec. To understand the role of Titan and non-Titan sources for H atoms in the Saturn system, the lifetime of hydrogen in the planetary magnetosphere was incorporated into the earlier Titan torus model of Smyth (1981) and its expected impact discussed. A particle trajectory model for cometary hydrogen is presented and applied to the Lyman-alpha distribution of Comet Kohoutek (1973XII).

Fuel Flexible Gas Turbine Combustor Flametube Facility Upgraded

NASA Technical Reports Server (NTRS)

Little, James E.; Nemets, Steve A.; Tornabene, Robert T.; Smith, Timothy D.; Frankenfeld, Bruce J.

2004-01-01

In fiscal year 2003, test cell 23 of the Research Combustion Laboratory (RCL 23) at the NASA Glenn Research Center was upgraded with the addition of gaseous hydrogen as a working propellant and the addition of a 450-psig air-supply system. Test flexibility was further enhanced by upgrades to the facility control systems. RCL 23 can now test with gaseous hydrogen flow rates up to 0.05 lbm/sec and jet fuel flow rates up to 0.62 lbm/sec. Research airflow rates up to 3 lbm/sec are possible with the 450-psig supply system over a range of inlet temperatures. Nonvitiated, heated air is supplied from a shell and tube heat exchanger. The maximum nonvitiated facility air temperature is 1100 F at 1.5 lbm/sec. Research-section exhaust temperatures are limited to 3200 F because of material and cooling capacity limits. A variety of support systems are available depending on the research hardware configuration. Test section ignition can be provided via either a hydrogen air torch system or an electronic spark system. Emissions measurements are obtained with either pneumatically or electromechanically actuated gas sample probes, and the electromechanical system allows for radial measurements at a user-specified axial location for measurement of emissions profiles. Gas analysis data can be obtained for a variety of species, including carbon monoxide (CO), carbon dioxide (CO2), nitrogen oxides (NO and NOx), oxygen (O2), unburnt hydrocarbons, and unburnt hydrogen. Facility control is accomplished with a programmable logic control system. Facility operations have been upgraded to a system based on graphical user interface control screens. A data system is available for real-time acquisition and monitoring of both measurements in engineering units and performance calculations. The upgrades have made RCL 23 a highly flexible facility for research into low emissions gas turbine combustor concepts, and the flame tube configuration inherently allows for a variety of fuel nozzle configurations to be tested in a cost-effective manner. RCL 23 is poised to be a leading facility for developing modern low-emission fuel nozzles for use with jet fuel and alternative fuels.

Two alternative binding mechanisms connect the protein translocation Sec71-Sec72 complex with heat shock proteins.

PubMed

Tripathi, Arati; Mandon, Elisabet C; Gilmore, Reid; Rapoport, Tom A

2017-05-12

The biosynthesis of many eukaryotic proteins requires accurate targeting to and translocation across the endoplasmic reticulum membrane. Post-translational protein translocation in yeast requires both the Sec61 translocation channel, and a complex of four additional proteins: Sec63, Sec62, Sec71, and Sec72. The structure and function of these proteins are largely unknown. This pathway also requires the cytosolic Hsp70 protein Ssa1, but whether Ssa1 associates with the translocation machinery to target protein substrates to the membrane is unclear. Here, we use a combined structural and biochemical approach to explore the role of Sec71-Sec72 subcomplex in post-translational protein translocation. To this end, we report a crystal structure of the Sec71-Sec72 complex, which revealed that Sec72 contains a tetratricopeptide repeat (TPR) domain that is anchored to the endoplasmic reticulum membrane by Sec71. We also determined the crystal structure of this TPR domain with a C-terminal peptide derived from Ssa1, which suggests how Sec72 interacts with full-length Ssa1. Surprisingly, Ssb1, a cytoplasmic Hsp70 that binds ribosome-associated nascent polypeptide chains, also binds to the TPR domain of Sec72, even though it lacks the TPR-binding C-terminal residues of Ssa1. We demonstrate that Ssb1 binds through its ATPase domain to the TPR domain, an interaction that leads to inhibition of nucleotide exchange. Taken together, our results suggest that translocation substrates can be recruited to the Sec71-Sec72 complex either post-translationally through Ssa1 or co-translationally through Ssb1. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Nascent chain-monitored remodeling of the Sec machinery for salinity adaptation of marine bacteria

PubMed Central

Ishii, Eiji; Chiba, Shinobu; Hashimoto, Narimasa; Kojima, Seiji; Homma, Michio; Ito, Koreaki; Akiyama, Yoshinori; Mori, Hiroyuki

2015-01-01

SecDF interacts with the SecYEG translocon in bacteria and enhances protein export in a proton-motive-force-dependent manner. Vibrio alginolyticus, a marine-estuarine bacterium, contains two SecDF paralogs, V.SecDF1 and V.SecDF2. Here, we show that the export-enhancing function of V.SecDF1 requires Na+ instead of H+, whereas V.SecDF2 is Na+-independent, presumably requiring H+. In accord with the cation-preference difference, V.SecDF2 was only expressed under limited Na+ concentrations whereas V.SecDF1 was constitutive. However, it is not the decreased concentration of Na+ per se that the bacterium senses to up-regulate the V.SecDF2 expression, because marked up-regulation of the V.SecDF2 synthesis was observed irrespective of Na+ concentrations under certain genetic/physiological conditions: (i) when the secDF1VA gene was deleted and (ii) whenever the Sec export machinery was inhibited. VemP (Vibrio export monitoring polypeptide), a secretory polypeptide encoded by the upstream ORF of secDF2VA, plays the primary role in this regulation by undergoing regulated translational elongation arrest, which leads to unfolding of the Shine–Dalgarno sequence for translation of secDF2VA. Genetic analysis of V. alginolyticus established that the VemP-mediated regulation of SecDF2 is essential for the survival of this marine bacterium in low-salinity environments. These results reveal that a class of marine bacteria exploits nascent-chain ribosome interactions to optimize their protein export pathways to propagate efficiently under different ionic environments that they face in their life cycles. PMID:26392525

Two alternative binding mechanisms connect the protein translocation Sec71-Sec72 complex with heat shock proteins

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

Tripathi, Arati; Mandon, Elisabet C.; Gilmore, Reid

The biosynthesis of many eukaryotic proteins requires accurate targeting to and translocation across the endoplasmic reticulum membrane. Post-translational protein translocation in yeast requires both the Sec61 translocation channel, and a complex of four additional proteins: Sec63, Sec62, Sec71, and Sec72. The structure and function of these proteins are largely unknown. This pathway also requires the cytosolic Hsp70 protein Ssa1, but whether Ssa1 associates with the translocation machinery to target protein substrates to the membrane is unclear. Here, we use a combined structural and biochemical approach to explore the role of Sec71-Sec72 subcomplex in post-translational protein translocation. To this end, wemore » report a crystal structure of the Sec71-Sec72 complex, which revealed that Sec72 contains a tetratricopeptide repeat (TPR) domain that is anchored to the endoplasmic reticulum membrane by Sec71. We also determined the crystal structure of this TPR domain with a C-terminal peptide derived from Ssa1, which suggests how Sec72 interacts with full-length Ssa1. Surprisingly, Ssb1, a cytoplasmic Hsp70 that binds ribosome-associated nascent polypeptide chains, also binds to the TPR domain of Sec72, even though it lacks the TPR-binding C-terminal residues of Ssa1. We demonstrate that Ssb1 binds through its ATPase domain to the TPR domain, an interaction that leads to inhibition of nucleotide exchange. Taken together, our results suggest that translocation substrates can be recruited to the Sec71-Sec72 complex either post-translationally through Ssa1 or co-translationally through Ssb1.« less

Radiation-free distal locking of intramedullary nails: evaluation of a new electromagnetic computer-assisted guidance system.

PubMed

Stathopoulos, Ioannis; Karampinas, Panagiotis; Evangelopoulos, Dimitrios-Stergios; Lampropoulou-Adamidou, Kalliopi; Vlamis, John

2013-06-01

Distal locking of intramedullary nails (IMNs) is a difficult part of intramedullary nailing (IMN) that could be time-consuming and expose the surgeon, the surgery personnel and the patient to a considerable amount of radiation as fluoroscopy is usually guiding the procedure. Utilization of electromagnetic fields for that purpose offers an attractive alternative. The SURESHOT™ Distal Targeting System (Smith & Nephew, Inc., Memphis, TN, USA) is a novel commercially available radiation-free aiming system that utilizes computerized electromagnetic field tracking technology for the distal locking of IMNs. In order to evaluate the efficacy of the system we conducted the present study. Nineteen patients (six females-thirteen males, mean age 39.5 years, range 17-85 years) with closed diaphyseal fracture of the femur (eight patients) or the tibia (eleven patients) were treated with IMN using the SURESHOT™ Distal Targeting System for the distal interlocking. All targeting attempts were successful at first try and followed by correct positioning of the screws. Mean time for distal locking of tibial IMNs (two screws) was 219sec (range 200-250sec). Mean time for distal locking of femoral IMNs (two screws) was 249 (range 220-330sec). In the current study the SURESHOT™ Distal Targeting System proved to be accurate, fast and easy to learn. Copyright © 2012 Elsevier Ltd. All rights reserved.

Gliding Motility and Por Secretion System Genes Are Widespread among Members of the Phylum Bacteroidetes

PubMed Central

Zhu, Yongtao

2013-01-01

The phylum Bacteroidetes is large and diverse, with rapid gliding motility and the ability to digest macromolecules associated with many genera and species. Recently, a novel protein secretion system, the Por secretion system (PorSS), was identified in two members of the phylum, the gliding bacterium Flavobacterium johnsoniae and the nonmotile oral pathogen Porphyromonas gingivalis. The components of the PorSS are not similar in sequence to those of other well-studied bacterial secretion systems. The F. johnsoniae PorSS genes are a subset of the gliding motility genes, suggesting a role for the secretion system in motility. The F. johnsoniae PorSS is needed for assembly of the gliding motility apparatus and for secretion of a chitinase, and the P. gingivalis PorSS is involved in secretion of gingipain protease virulence factors. Comparative analysis of 37 genomes of members of the phylum Bacteroidetes revealed the widespread occurrence of gliding motility genes and PorSS genes. Genes associated with other bacterial protein secretion systems were less common. The results suggest that gliding motility is more common than previously reported. Microscopic observations confirmed that organisms previously described as nonmotile, including Croceibacter atlanticus, “Gramella forsetii,” Paludibacter propionicigenes, Riemerella anatipestifer, and Robiginitalea biformata, exhibit gliding motility. Three genes (gldA, gldF, and gldG) that encode an apparent ATP-binding cassette transporter required for F. johnsoniae gliding were absent from two related gliding bacteria, suggesting that the transporter may not be central to gliding motility. PMID:23123910

Engineering Escherichia coli into a protein delivery system for mammalian cells.

PubMed

Reeves, Analise Z; Spears, William E; Du, Juan; Tan, Kah Yong; Wagers, Amy J; Lesser, Cammie F

2015-05-15

Many Gram-negative pathogens encode type 3 secretion systems, sophisticated nanomachines that deliver proteins directly into the cytoplasm of mammalian cells. These systems present attractive opportunities for therapeutic protein delivery applications; however, their utility has been limited by their inherent pathogenicity. Here, we report the reengineering of a laboratory strain of Escherichia coli with a tunable type 3 secretion system that can efficiently deliver heterologous proteins into mammalian cells, thereby circumventing the need for virulence attenuation. We first introduced a 31 kB region of Shigella flexneri DNA that encodes all of the information needed to form the secretion nanomachine onto a plasmid that can be directly propagated within E. coli or integrated into the E. coli chromosome. To provide flexible control over type 3 secretion and protein delivery, we generated plasmids expressing master regulators of the type 3 system from either constitutive or inducible promoters. We then constructed a Gateway-compatible plasmid library of type 3 secretion sequences to enable rapid screening and identification of sequences that do not perturb function when fused to heterologous protein substrates and optimized their delivery into mammalian cells. Combining these elements, we found that coordinated expression of the type 3 secretion system and modified target protein substrates produces a nonpathogenic strain that expresses, secretes, and delivers heterologous proteins into mammalian cells. This reengineered system thus provides a highly flexible protein delivery platform with potential for future therapeutic applications.

A "push and slide" mechanism allows sequence-insensitive translocation of secretory proteins by the SecA ATPase.

PubMed

Bauer, Benedikt W; Shemesh, Tom; Chen, Yu; Rapoport, Tom A

2014-06-05

In bacteria, most secretory proteins are translocated across the plasma membrane by the interplay of the SecA ATPase and the SecY channel. How SecA moves a broad range of polypeptide substrates is only poorly understood. Here we show that SecA moves polypeptides through the SecY channel by a "push and slide" mechanism. In its ATP-bound state, SecA interacts through a two-helix finger with a subset of amino acids in a substrate, pushing them into the channel. A polypeptide can also passively slide back and forth when SecA is in the predominant ADP-bound state or when SecA encounters a poorly interacting amino acid in its ATP-bound state. SecA performs multiple rounds of ATP hydrolysis before dissociating from SecY. The proposed push and slide mechanism is supported by a mathematical model and explains how SecA allows translocation of a wide range of polypeptides. This mechanism may also apply to hexameric polypeptide-translocating ATPases. Copyright © 2014 Elsevier Inc. All rights reserved.

Demonstration of a Specific Escherichia coli SecY–Signal Peptide Interaction†

PubMed Central

Wang, Ligong; Miller, Alexander; Rusch, Sharyn L.; Kendall, Debra A.

2011-01-01

Protein translocation in Escherichia coli is initiated by the interaction of a preprotein with the membrane translocase composed of a motor protein, SecA ATPase, and a membrane-embedded channel, the SecYEG complex. The extent to which the signal peptide region of the preprotein plays a role in SecYEG interactions is unclear, in part because studies in this area typically employ the entire preprotein. Using a synthetic signal peptide harboring a photoaffinity label in its hydrophobic core, we examined this interaction with SecYEG in a detergent micellar environment. The signal peptide was found to specifically bind SecY in a saturable manner and at levels comparable to those that stimulate SecA ATPase activity. Chemical and proteolytic cleavage of cross-linked SecY and analysis of the signal peptide adducts indicate that the binding was primarily to regions of the protein containing transmembrane domains seven and two. The signal peptide–SecY interaction was affected by the presence of SecA and nucleotides in a manner consistent with the transfer of signal peptide to SecY upon nucleotide hydrolysis at SecA. PMID:15476412

Analysis of a waterborne disease model with socioeconomic classes.

PubMed

Collins, O C; Robertson, Suzanne L; Govinder, K S

2015-11-01

Waterborne diseases such as cholera continue to pose serious public health problems in the world today. Transmission parameters can vary greatly with socioeconomic class (SEC) and the availability of clean water. We formulate a multi-patch waterborne disease model such that each patch represents a particular SEC with its own water source, allowing individuals to move between SECs. For a 2-SEC model, we investigate the conditions under which each SEC is responsible for driving a cholera outbreak. We determine the effect of SECs on disease transmission dynamics by comparing the basic reproduction number of the 2-SEC model to that of a homogeneous model that does not take SECs into account. We conclude by extending several results of the 2-SEC model to an n-SEC model. Copyright © 2015 Elsevier Inc. All rights reserved.

14 CFR Sec. 1-2 - Waivers from this system of accounts and reports.

Code of Federal Regulations, 2010 CFR

2010-01-01

... (AVIATION PROCEEDINGS) ECONOMIC REGULATIONS UNIFORM SYSTEM OF ACCOUNTS AND REPORTS FOR LARGE CERTIFICATED... the principles embodied in the provisions of this system of accounts and reports; and the application...

The Effects of Increasing Ocular Surface Stimulation on Blinking and Tear Secretion

PubMed Central

Wu, Ziwei; Begley, Carolyn G.; Port, Nicholas; Bradley, Arthur; Braun, Richard; King-Smith, Ewen

2015-01-01

Purpose. To investigate the effect of varying levels of ocular surface stimulation on the timing and amplitude of the blink and tear secretion. Methods. Following instillation of fluorescein dye, increasing levels of air flow were directed toward the central corneas of 10 healthy subjects. Interblink interval (IBI), tear meniscus height (TMH), and fluorescence intensity were measured simultaneously. Because blinking can obscure changes in TMH, we developed novel measures of tear secretion by calculating tear meniscus fluorescein concentration (TMFC) from intensity using a mathematical model. The change of TMH and TMFC over trials and the slope of the TMFC within each IBI (IBI-TTR) were further calculated. Results. The mean IBI was decreased by 8.08 ± 8.54 seconds from baseline to maximum air stimulation. The TMH increase was highly variable (0.41 ± 0.39 mm) among subjects, compared to the fluorescence tear turnover metrics: decrease in TMFC of 2.84 ± 0.98 natural logarithm or ln(%) and IBI-TTR of 0.065 ± 0.032 ln(%)/sec. Ocular surface stimulation was highly correlated with the TMFC and IBI-TTR, but less so with TMH (Pearson's r = 0.71, 0.69, and 0.40, P < 0.01, respectively). Blinking and tearing were significantly correlated with each other (Pearson's r = 0.56, P < 0.01), but tearing lagged behind by an average of 6.54 ± 4.07 seconds. Conclusions. Blinking and tearing share a common origin with sensory stimulation at the ocular surface. Both showed a dose–response increase with surface stimulation and were correlated with each other. These methods can potentially be used to understand alterations in ocular surface sensory function and associated protective responses in dry eye and other disorders of the ocular surface. PMID:26132780

The Effects of Increasing Ocular Surface Stimulation on Blinking and Tear Secretion.

PubMed

Wu, Ziwei; Begley, Carolyn G; Port, Nicholas; Bradley, Arthur; Braun, Richard; King-Smith, Ewen

2015-07-01

To investigate the effect of varying levels of ocular surface stimulation on the timing and amplitude of the blink and tear secretion. Following instillation of fluorescein dye, increasing levels of air flow were directed toward the central corneas of 10 healthy subjects. Interblink interval (IBI), tear meniscus height (TMH), and fluorescence intensity were measured simultaneously. Because blinking can obscure changes in TMH, we developed novel measures of tear secretion by calculating tear meniscus fluorescein concentration (TMFC) from intensity using a mathematical model. The change of TMH and TMFC over trials and the slope of the TMFC within each IBI (IBI-TTR) were further calculated. The mean IBI was decreased by 8.08 ± 8.54 seconds from baseline to maximum air stimulation. The TMH increase was highly variable (0.41 ± 0.39 mm) among subjects, compared to the fluorescence tear turnover metrics: decrease in TMFC of 2.84 ± 0.98 natural logarithm or ln(%) and IBI-TTR of 0.065 ± 0.032 ln(%)/sec. Ocular surface stimulation was highly correlated with the TMFC and IBI-TTR, but less so with TMH (Pearson's r = 0.71, 0.69, and 0.40, P < 0.01, respectively). Blinking and tearing were significantly correlated with each other (Pearson's r = 0.56, P < 0.01), but tearing lagged behind by an average of 6.54 ± 4.07 seconds. Blinking and tearing share a common origin with sensory stimulation at the ocular surface. Both showed a dose-response increase with surface stimulation and were correlated with each other. These methods can potentially be used to understand alterations in ocular surface sensory function and associated protective responses in dry eye and other disorders of the ocular surface.

Escherichia coli type III secretion system 2: a new kind of T3SS?

PubMed

Zhou, Mingxu; Guo, Zhiyan; Duan, Qiangde; Hardwidge, Philip R; Zhu, Guoqiang

2014-03-19

Type III secretion systems (T3SSs) are employed by Gram-negative bacteria to deliver effector proteins into the cytoplasm of infected host cells. Enteropathogenic Escherichia coli use a T3SS to deliver effector proteins that result in the creation of the attaching and effacing lesions. The genome sequence of the Escherichia coli pathotype O157:H7 revealed the existence of a gene cluster encoding components of a second type III secretion system, the E. coli type III secretion system 2 (ETT2). Researchers have revealed that, although ETT2 may not be a functional secretion system in most (or all) strains, it still plays an important role in bacterial virulence. This article summarizes current knowledge regarding the E. coli ETT2, including its genetic characteristics, prevalence, function, association with virulence, and prospects for future work.

An improvement of the Earthworm Based Earthquake Alarm Reporting system in Taiwan

NASA Astrophysics Data System (ADS)

Chen, D. Y.; Hsiao, N. C.; Yih-Min, W.

2017-12-01

The Central Weather Bureau of Taiwan (CWB) has operated the Earthworm Based Earthquake Alarm Reporting (eBEAR) system for the purpose of earthquake early warning (EEW). The system has been used to report EEW messages to the general public since 2016 through text message from the mobile phones and the television programs. The system for inland earthquakes is able to provide accurate and fast warnings. The average epicenter error is about 5 km and the processing time is about 15 seconds. The epicenter error is defined as the distance between the epicenter estimated by the EEW system and the epicenter estimated by man. The processing time is defined as the time difference between the time earthquakes occurred and the time the system issued warning. The CWB seismic network consist about 200 seismic stations. In some area of Taiwan the distance between each seismic station is about 10 km. It means that when an earthquake occurred the seismic P wave is able to propagate through 6 stations, which is the minimum number of required stations in the EEW system, within 20 km. If the latency of data transmitting is about 1 sec, the P-wave velocity is about 6 km per sec and we take 3-sec length time window to estimate earthquake magnitude, then the processing should be around 8 sec. In fact, however, the average processing time is larger than this figure. Because some outliers of P-wave onset picks may exist in the beginning of the earthquake occurrence, the Geiger's method we used in the EEW system for earthquake location is not stable. It usually takes more time to wait for enough number of good picks. In this study we used grid search method to improve the estimations of earthquake location. The MAXEL algorithm (Sheen et al., 2015, 2016) was tested in the EEW system by simulating historical earthquakes occurred in Taiwan. The results show the processing time can be reduced and the location accuracy is acceptable for EEW purpose.

Environmental Assessment: Demolish CASS Switch Stations Buildings 644, 645, 646 at Grand Forks Air Force Base

DTIC Science & Technology

2006-06-01

Per Year TSCA Toxic Substance Control Act TSI Thermal System Insulation UAV Unmanned Aerial Vehicle UHF Ultra High Frequency UPS Uninterruptible...Conservation and Recovery Act (RCRA) of 1976 [42 U.S.C. Sec. 6901, et seq.] • Toxic Substances Control Act (TSCA) of 1976 [15 U.S.C. Sec. 2601, et...Corporation (SAIC). Typical hazardous materials include reactive materials such as explosives, ignitables, toxics , and corrosives. Improper storage can

System Control for the Transitional DCS. Appendices.

DTIC Science & Technology

1978-12-01

the deployment of the AN/TTC-39 circuit switch. This is a hybrid analog/digital switch providing the following services: o Non- secure analog telephone...service. o Non- secure 16 Kb/s digital telephone service. o Secure 16 Kb/s digital telephone service with automatic key distribution and end to end... security . o Analog circuits to support current inventory 50 Kb/sec and 9.6 Kb/sec secure digital communications. In the deployment model for this study

A Power Hardware-in-the-Loop Platform with Remote Distribution Circuit Cosimulation

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

Palmintier, Bryan; Lundstrom, Blake; Chakraborty, Sudipta

2015-04-01

This paper demonstrates the use of a novel cosimulation architecture that integrates hardware testing using Power Hardware-in-the-Loop (PHIL) with larger-scale electric grid models using off-the-shelf, non-PHIL software tools. This architecture enables utilities to study the impacts of emerging energy technologies on their system and manufacturers to explore the interactions of new devices with existing and emerging devices on the power system, both without the need to convert existing grid models to a new platform or to conduct in-field trials. The paper describes an implementation of this architecture for testing two residential-scale advanced solar inverters at separate points of common coupling.more » The same hardware setup is tested with two different distribution feeders (IEEE 123 and 8500 node test systems) modeled using GridLAB-D. In addition to simplifying testing with multiple feeders, the architecture demonstrates additional flexibility with hardware testing in one location linked via the Internet to software modeling in a remote location. In testing, inverter current, real and reactive power, and PCC voltage are well captured by the co-simulation platform. Testing of the inverter advanced control features is currently somewhat limited by the software model time step (1 sec) and tested communication latency (24 msec). Overshoot induced oscillations are observed with volt/VAR control delays of 0 and 1.5 sec, while 3.4 sec and 5.5 sec delays produced little or no oscillation. These limitations could be overcome using faster modeling and communication within the same co-simulation architecture.« less

Optimization of stent implantation using a high pressure inflation protocol.

PubMed

Vallurupalli, Srikanth; Bahia, Amit; Ruiz-Rodriguez, Ernesto; Ahmed, Zubair; Hakeem, Abdul; Uretsky, Barry F

2016-01-01

High-pressure inflation is the universal standard for stent deployment but a specific protocol for its use is lacking. We developed a standardized "pressure optimization protocol" (POP) using time to inflation pressure stability as an endpoint for determining the required duration of stent inflation. The primary study purpose was to determine the stent inflation time (IT) in a large patient cohort using the standardized inflation protocol, to correlate various patient and lesion characteristics with IT, and ascertain in an in vitro study the time for pressure accommodation within an inflation system. Six hundred fifteen stent implants in 435 patients were studied. Multivariate analysis was performed to determine predictors of longer ITs. In an in vitro study, various stents and balloons were inflated in air to determine the pressure accommodation time of the inflation system. The mean stent IT was 104 ± 41 sec (range 30-380 sec). Stent length was the only predictor of prolonged stent inflation. The "accommodation time" in vitro of the stent inflation system itself was 33 ± 24 sec. The protocol was safe requiring premature inflation termination in <3% of stent implants. No serious adverse events occurred. Achieving stable inflation pressure requires on average over 100 sec and may require several minutes in individual cases. Stent length increases IT. These results suggest that the widespread practice of rapid inflation/deflation may not be sufficient to fully expand the stent and that the use of a pressure stability protocol will allow for safe, predictable, and more complete stent deployment. © 2015 Wiley Periodicals, Inc. © 2015 Wiley Periodicals, Inc.

Fermilab`s DART DA system

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

Pordes, R.; Anderson, J.; Berg, D.

1994-04-01

DART is the new data acquisition system designed and implemented for six Fermilab experiments by the Fermilab Computing Division and the experiments themselves. The complexity of the experiments varies greatly. Their data taking throughput and event filtering requirements range from a few (2-5) to tens (80) of CAMAC, FASTBUS and home built front end crates; from a few 100 KByte/sec to 160 MByte/sec front end data collection rates; and from 0-3000 Mips of level 3 processing. The authors report on the architecture and implementation of DART to this date, and the hardware and software components that are being developed andmore » supported.« less

Fermilab`s DART DA system

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

Pordes, R.; Anderson, J.; Berg, D.

1994-12-31

DART is the new data acquisition system designed and implemented for six Fermilab experiments by the Fermilab Computing Division and the experiments themselves. The complexity of the experiments varies greatly. Their data taking throughput and event filtering requirements range from a few (2-5) to tens (80) of CAMAC, FASTBUS and home built front end crates; from a few 100 KByte/sec to 160 MByte/sec front end data collection rates; and from 0-3000 Mips of level 3 processing. The authors report on the architecture and implementation of DART to this data, and the hardware and software components that are being developed andmore » supported.« less

Extreme Ultraviolet Solar Images Televised In-Flight with a Rocket-Borne SEC Vidicon System.

PubMed

Tousey, R; Limansky, I

1972-05-01

A TV image of the entire sun while an importance 2N solar flare was in progress was recorded in the extreme ultraviolet (XUV) radiation band 171-630 A and transmitted to ground from an Aerobee-150 rocket on 4 November 1969 using S-band telemetry. The camera tube was a Westinghouse Electric Corporation SEC vidicon, with its fiber optic faceplate coated with an XUV to visible conversion layer of p-quaterphenyl. The XUV passband was produced by three 1000-A thick aluminum filters in series together with the platinized reflecting surface of the off-axis paraboloid that imaged the sun. A number of images were recorded with integration times between 1/30 see and 2 sec. Reconstruction of pictures was enhanced by combining several to reduce the noise.

A Dwarf Dissolving? - A Kinematic Analysis of Andromeda XXVII and the Northern Arc

NASA Astrophysics Data System (ADS)

Collins, Michelle; Rich, R. M.; Chapman, S. C.; Ibata, R.; Irwin, M.; McConnachie, A. W.

2013-01-01

We report internal kinematics for an unusual M31 dwarf spheroidal galaxy, And XXVII, which is superposed against the Northern Arc Stream feature, isolated in the PandAS (Pan-Andromeda Archaeological Survey). In contrast to the coherent, cold velocity fields of most Andromeda dwarf spheroidals, And XXVII has a trimodal velocity distribution spanning 100 km/sec, with a relatively cold central peak at -530 km/sec , and a velocity dispersion of sigma= 8 km/sec. While all of the candidate members are < 2' (or approximately one half light radii, ~600 pc) from the core, the full velocity range is not consistent with a system of luminosity Mv=-7.9. We propose that And XXVII may be in the process of dissolving into the Northern Arc.

Leak detection with expandable coatings

NASA Technical Reports Server (NTRS)

1971-01-01

Developed and evaluated is a system for leak detection that can be easily applied over separable connectors and that expands into a bubble or balloon if a leak is present. This objective is accomplished by using thin films of Parafilm tape wrapped over connectors, which are then overcoated with a special formulation. The low yield strength and the high elongation of the envelope permit bubble formation if leakage occurs. This system is appropriate for welds and other hardware besides separable connectors. The practical limit of this system appears to be for leaks exceeding 0.000001 cc/sec. If this envelope is used to trap gases for mass spectrometer inspection, leaks in the range of ten to the minus 8th power cc/sec. may be detectable.

Dynamic Glycosylation Governs the Vertebrate COPII Protein Trafficking Pathway.

PubMed

Cox, Nathan J; Unlu, Gokhan; Bisnett, Brittany J; Meister, Thomas R; Condon, Brett M; Luo, Peter M; Smith, Timothy J; Hanna, Michael; Chhetri, Abhishek; Soderblom, Erik J; Audhya, Anjon; Knapik, Ela W; Boyce, Michael

2018-01-09

The COPII coat complex, which mediates secretory cargo trafficking from the endoplasmic reticulum, is a key control point for subcellular protein targeting. Because misdirected proteins cannot function, protein sorting by COPII is critical for establishing and maintaining normal cell and tissue homeostasis. Indeed, mutations in COPII genes cause a range of human pathologies, including cranio-lenticulo-sutural dysplasia (CLSD), which is characterized by collagen trafficking defects, craniofacial abnormalities, and skeletal dysmorphology. Detailed knowledge of the COPII pathway is required to understand its role in normal cell physiology and to devise new treatments for disorders in which it is disrupted. However, little is known about how vertebrates dynamically regulate COPII activity in response to developmental, metabolic, or pathological cues. Several COPII proteins are modified by O-linked β-N-acetylglucosamine (O-GlcNAc), a dynamic form of intracellular protein glycosylation, but the biochemical and functional effects of these modifications remain unclear. Here, we use a combination of chemical, biochemical, cellular, and genetic approaches to demonstrate that site-specific O-GlcNAcylation of COPII proteins mediates their protein-protein interactions and modulates cargo secretion. In particular, we show that individual O-GlcNAcylation sites of SEC23A, an essential COPII component, are required for its function in human cells and vertebrate development, because mutation of these sites impairs SEC23A-dependent in vivo collagen trafficking and skeletogenesis in a zebrafish model of CLSD. Our results indicate that O-GlcNAc is a conserved and critical regulatory modification in the vertebrate COPII-dependent trafficking pathway.

Differentiation and classification of phytoplasmas in the pigeon pea witches'-broom group (16SrIX): an update based on multiple gene sequence analysis.

PubMed

Lee, I-M; Bottner-Parker, K D; Zhao, Y; Bertaccini, A; Davis, R E

2012-09-01

The pigeon pea witches'-broom phytoplasma group (16SrIX) comprises diverse strains that cause numerous diseases in leguminous trees and herbaceous crops, vegetables, a fruit, a nut tree and a forest tree. At least 14 strains have been reported worldwide. Comparative phylogenetic analyses of the highly conserved 16S rRNA gene and the moderately conserved rplV (rpl22)-rpsC (rps3) and secY genes indicated that the 16SrIX group consists of at least six distinct genetic lineages. Some of these lineages cannot be readily differentiated based on analysis of 16S rRNA gene sequences alone. The relative genetic distances among these closely related lineages were better assessed by including more variable genes [e.g. ribosomal protein (rp) and secY genes]. The present study demonstrated that virtual RFLP analyses using rp and secY gene sequences allowed unambiguous identification of such lineages. A coding system is proposed to designate each distinct rp and secY subgroup in the 16SrIX group.

Authentication Without Secrets

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

Pierson, Lyndon G.; Robertson, Perry J.

This work examines a new approach to authentication, which is the most fundamental security primitive that underpins all cyber security protections. Current Internet authentication techniques require the protection of one or more secret keys along with the integrity protection of the algorithms/computations designed to prove possession of the secret without actually revealing it. Protecting a secret requires physical barriers or encryption with yet another secret key. The reason to strive for "Authentication without Secret Keys" is that protecting secrets (even small ones only kept in a small corner of a component or device) is much harder than protecting the integritymore » of information that is not secret. Promising methods are examined for authentication of components, data, programs, network transactions, and/or individuals. The successful development of authentication without secret keys will enable far more tractable system security engineering for high exposure, high consequence systems by eliminating the need for brittle protection mechanisms to protect secret keys (such as are now protected in smart cards, etc.). This paper is a re-release of SAND2009-7032 with new figures numerous edits.« less

Dynamic distribution of the SecA and SecY translocase subunits and septal localization of the HtrA surface chaperone/protease during Streptococcus pneumoniae D39 cell division.

PubMed

Tsui, Ho-Ching Tiffany; Keen, Susan K; Sham, Lok-To; Wayne, Kyle J; Winkler, Malcolm E

2011-01-01

The Sec translocase pathway is the major route for protein transport across and into the cytoplasmic membrane of bacteria. Previous studies reported that the SecA translocase ATP-binding subunit and the cell surface HtrA protease/chaperone formed a single microdomain, termed "ExPortal," in some species of ellipsoidal (ovococcus) Gram-positive bacteria, including Streptococcus pyogenes. To investigate the generality of microdomain formation, we determined the distribution of SecA and SecY by immunofluorescent microscopy in Streptococcus pneumoniae (pneumococcus), which is an ovococcus species evolutionarily distant from S. pyogenes. In the majority (≥ 75%) of exponentially growing cells, S. pneumoniae SecA (SecA (Spn)) and SecY (Spn) located dynamically in cells at different stages of division. In early divisional cells, both Sec subunits concentrated at equators, which are future sites of constriction. Further along in division, SecA(Spn) and SecY(Spn) remained localized at mid-cell septa. In late divisional cells, both Sec subunits were hemispherically distributed in the regions between septa and the future equators of dividing cells. In contrast, the HtrA (Spn) homologue localized to the equators and septa of most (> 90%) dividing cells, whereas the SrtA(Spn) sortase located over the surface of cells in no discernable pattern. This dynamic pattern of Sec distribution was not perturbed by the absence of flotillin family proteins, but was largely absent in most cells in early stationary phase and in cls mutants lacking cardiolipin synthase. These results do not support the existence of an ExPortal microdomain in S. pneumoniae. Instead, the localization of the pneumococcal Sec translocase depends on the stage of cell division and anionic phospholipid content. Two patterns of Sec translocase distribution, an ExPortal microdomain in certain ovococcus-shaped species like Streptococcus pyogenes and a spiral pattern in rod-shaped species like Bacillus subtilis, have been reported for Gram-positive bacteria. This study provides evidence for a third pattern of Sec localization in the ovococcus human pathogen Streptococcus pneumoniae. The SecA motor and SecY channel subunits of the Sec translocase localize dynamically to different places in the mid-cell region during the division cycle of exponentially growing, but not stationary-phase, S. pneumoniae. Unexpectedly, the S. pneumoniae HtrA (HtrA(Spn)) protease/chaperone principally localizes to cell equators and division septa. The coincident localization of SecA(Spn), SecY (Spn), and HtrA (Spn) to regions of peptidoglycan (PG) biosynthesis in unstressed, growing cells suggests that the pneumococcal Sec translocase directs assembly of the PG biosynthesis apparatus to regions where it is needed during division and that HtrA(Spn) may play a general role in quality control of proteins exported by the Sec translocase.

NMDA inhibits oxotremorine-induced acid secretion via the NO-dependent cyclic GMP system in rat stomach.

PubMed

Tsai, L H; Lee, Y J

2001-12-31

The mechanism of N-methyl-D-aspartate (NMDA) inhibits oxotremorine-induced acid secretion was examined in rat stomach, in relation to the cyclic GMP system. NMDA (10(-7) M) did not affect the spontaneous acid secretion from the everted preparations of isolated rat stomach, but inhibited the acid secretion stimulated by oxotremorine, and this effect of NMDA was antagonized by 2-amino-5-phosphonovaleric acid (AP-5), (+/-)3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid (CPP) or N(G)-nitro-L-arginine (L-NNA). NMDA also elevated the cyclic GMP content of mucosal slices from rat stomach, and this effect of NMDA was antagonized by L-NNA. These results indicate that NMDA receptors are present in the rat stomach and regulate the gastric acid secretion. The mechanism underlying the effect of NMDA inhibits oxotremorine-induced acid secretion may be mediated by the NO-dependent cyclic GMP system.

An optical disk archive for a data base management system

NASA Technical Reports Server (NTRS)

Thomas, Douglas T.

1985-01-01

An overview is given of a data base management system that can catalog and archive data at rates up to 50M bits/sec. Emphasis is on the laser disk system that is used for the archive. All key components in the system (3 Vax 11/780s, a SEL 32/2750, a high speed communication interface, and the optical disk) are interfaced to a 100M bits/sec 16-port fiber optic bus to achieve the high data rates. The basic data unit is an autonomous data packet. Each packet contains a primary and secondary header and can be up to a million bits in length. The data packets are recorded on the optical disk at the same time the packet headers are being used by the relational data base management software ORACLE to create a directory independent of the packet recording process. The user then interfaces to the VAX that contains the directory for a quick-look scan or retrieval of the packet(s). The total system functions are distributed between the VAX and the SEL. The optical disk unit records the data with an argon laser at 100M bits/sec from its buffer, which is interfaced to the fiber optic bus. The same laser is used in the read cycle by reducing the laser power. Additional information is given in the form of outlines, charts, and diagrams.

Twin-arginine signal peptide of Bacillus subtilis YwbN can direct Tat-dependent secretion of methyl parathion hydrolase.

PubMed

Liu, Ruihua; Zuo, Zhenqiang; Xu, Yingming; Song, Cunjiang; Jiang, Hong; Qiao, Chuanling; Xu, Ping; Zhou, Qixing; Yang, Chao

2014-04-02

The twin-arginine translocation (Tat) pathway exports folded proteins across the cytoplasmic membranes of bacteria and archaea. Two parallel Tat pathways (TatAdCd and TatAyCy systems) with distinct substrate specificities have previously been discovered in Bacillus subtilis. In this study, to secrete methyl parathion hydrolase (MPH) into the growth medium, the twin-arginine signal peptide of B. subtilis YwbN was used to target MPH to the Tat pathway of B. subtilis. Western blot analysis and MPH assays demonstrated that active MPH was secreted into the culture supernatant of wild-type cells. No MPH secretion occurred in a total-tat2 mutant, indicating that the observed export in wild-type cells was mediated exclusively by the Tat pathway. Export was fully blocked in a tatAyCy mutant. In contrast, the tatAdCd mutant was still capable of secreting MPH. These results indicated that the MPH secretion directed by the YwbN signal peptide was specifically mediated by the TatAyCy system. The N-terminal sequence of secreted MPH was determined as AAPQVR, demonstrating that the YwbN signal peptide had been processed correctly. This is the first report of functional secretion of a heterologous protein via the B. subtilis TatAyCy system. This study highlights the potential of the TatAyCy system to be used for secretion of other heterologous proteins in B. subtilis.

76 FR 72245 - Implementation of Defense Trade Cooperation Treaties

Federal Register 2010, 2011, 2012, 2013, 2014

2011-11-22

... clarifying language to Sec. Sec. 123.9(c), (c)(1), and (c)(2); and adding the language of the current (c)(4) to (c)(3). New language pertaining to new Sec. Sec. 126.16 and 126.17 will comprise a new (c)(4). We... clarifying language to Sec. Sec. 123.9(e)(3) and (e)(4). We are adding Israel to the list of countries and...

40 CFR 172.3 - Scope of requirement.

Code of Federal Regulations, 2014 CFR

2014-07-01

....C. sec. 321(p) and sec. 355). (2) A “new animal drug” (21 U.S.C. sec. 321(w) and sec. 360(b)), or (3) An “animal feed” (21 U.S.C. sec. 321 (x)) containing a “new animal drug” (21 U.S.C. sec. 360(b)). (g... experimental use permit (EUP) is generally required for testing of any unregistered pesticide or any registered...

40 CFR 172.3 - Scope of requirement.

Code of Federal Regulations, 2013 CFR

2013-07-01

....C. sec. 321(p) and sec. 355). (2) A “new animal drug” (21 U.S.C. sec. 321(w) and sec. 360(b)), or (3) An “animal feed” (21 U.S.C. sec. 321 (x)) containing a “new animal drug” (21 U.S.C. sec. 360(b)). (g... experimental use permit (EUP) is generally required for testing of any unregistered pesticide or any registered...

40 CFR 172.3 - Scope of requirement.

Code of Federal Regulations, 2011 CFR

2011-07-01

....C. sec. 321(p) and sec. 355). (2) A “new animal drug” (21 U.S.C. sec. 321(w) and sec. 360(b)), or (3) An “animal feed” (21 U.S.C. sec. 321 (x)) containing a “new animal drug” (21 U.S.C. sec. 360(b)). (g... experimental use permit (EUP) is generally required for testing of any unregistered pesticide or any registered...

Lst1p and Sec24p Cooperate in Sorting of the Plasma Membrane Atpase into Copii Vesicles in Saccharomyces cerevisiae

PubMed Central

Shimoni, Yuval; Kurihara, Tatsuo; Ravazzola, Mariella; Amherdt, Mylène; Orci, Lelio; Schekman, Randy

2000-01-01

Formation of ER-derived protein transport vesicles requires three cytosolic components, a small GTPase, Sar1p, and two heterodimeric complexes, Sec23/24p and Sec13/31p, which comprise the COPII coat. We investigated the role of Lst1p, a Sec24p homologue, in cargo recruitment into COPII vesicles in Saccharomyces cerevisiae. A tagged version of Lst1p was purified and eluted as a heterodimer complexed with Sec23p comparable to the Sec23/24p heterodimer. We found that cytosol from an lst1-null strain supported the packaging of α-factor precursor into COPII vesicles but was deficient in the packaging of Pma1p, the essential plasma membrane ATPase. Supplementation of mutant cytosol with purified Sec23/Lst1p restored Pma1p packaging into the vesicles. When purified COPII components were used in the vesicle budding reaction, Pma1p packaging was optimal with a mixture of Sec23/24p and Sec23/Lst1p; Sec23/Lst1p did not replace Sec23/24p. Furthermore, Pma1p coimmunoprecipitated with Lst1p and Sec24p from vesicles. Vesicles formed with a mixture of Sec23/Lst1p and Sec23/24p were similar morphologically and in their buoyant density, but larger than normal COPII vesicles (87-nm vs. 75-nm diameter). Immunoelectronmicroscopic and biochemical studies revealed both Sec23/Lst1p and Sec23/24p on the membranes of the same vesicles. These results suggest that Lst1p and Sec24p cooperate in the packaging of Pma1p and support the view that biosynthetic precursors of plasma membrane proteins must be sorted into ER-derived transport vesicles. Sec24p homologues may comprise a more complex coat whose combinatorial subunit composition serves to expand the range of cargo to be packaged into COPII vesicles. By changing the geometry of COPII coat polymerization, Lst1p may allow the transport of bulky cargo molecules, polymers, or particles. PMID:11086000

SU-E-J-273: Skin Temperature Recovery Rate as a Potential Predictor for Radiation-Induced Skin Reactions

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

Biswal, N C; Wu, Z; Chu, J

Purpose: To assess the potential of dynamic infrared imaging to evaluate early skin reactions during radiation therapy in cancer patients. Methods: Thermal images were captured by our home-built system consisting of two flash lamps and an infrared (IR) camera. The surface temperature of the skin was first raised by ∼ 6 °C from ∼1 ms short flashes; the camera then captured a series of IR images for 10 seconds. For each image series, a basal temperature was recorded for 0.5 seconds before flash was triggered. The temperature gradients (ε) were calculated between a reference point (immediately after the flash) andmore » at a time point of 2sec, 4sec and 9sec after that. A 1.0 cm region of interest (ROI) on the skin was drawn; the mean and standard deviations of the ROIs were calculated. The standard ε values for normal human skins were evaluated by imaging 3 healthy subjects with different skin colors. All of them were imaged on 3 separate days for consistency checks. Results: The temperature gradient, which is the temperature recovery rate, depends on the thermal properties of underlying tissue, i.e. thermal conductivity. The average ε for three volunteers averaged over 3 measurements were 0.64±0.1, 0.72±0.2 and 0.80±0.3 at 2sec, 4sec and 9sec respectively. The standard deviations were within 1.5%–3.2%. One of the volunteers had a prior small skin burn on the left wrist and the ε values for the burned site were around 9% (at 4sec) and 13% (at 9sec) lower than that from the nearby normal skin. Conclusion: The temperature gradients from the healthy subjects were reproducible within 1.5%–3.2 % and that from a burned skin showed a significant difference (9%–13%) from the normal skin. We have an IRB approved protocol to image head and neck patients scheduled for radiation therapy.« less

Comparison of lightning activity in the two most active areas of the Congo Basin

NASA Astrophysics Data System (ADS)

Kigotsi, Jean K.; Soula, Serge; Georgis, Jean-François

2018-02-01

A comparison of the lightning activity in the two most active areas (Area_max for the main maximum and Area_sec for the secondary maximum) of the Congo Basin is made with data obtained by the World Wide Lightning Location Network (WWLLN) during 2012 and 2013. Both areas of same size (5° × 5°) exhibit flash counts in a ratio of about 1.32 for both years and very different distributions of the flash rate density (FRD) with maximums in a ratio of 1.94 and 2.59 for 2012 and 2013, respectively. The FRD is much more widely distributed in Area_sec, which means the whole area contributes more or less equal to the lightning activity. The diurnal cycle is much more pronounced in Area_max than in Area_sec with a ratio between the maximum and the minimum of 15.4 and 4.7, respectively. However, the minimum and maximum of the hourly flash rates are observed roughly at the same time in both areas, between 07:00 and 09:00 UTC and between 16:00 and 17:00 UTC, respectively. In Area_sec the proportion of days with low lightning rate (0-1000 flashes per day) is much larger (˜ 45 % in 2013) compared to Area_max (˜ 23 % in 2013). In Area_max the proportion of days with moderate lightning rate (1001-6000 flashes per day) is larger (˜ 68.5 % in 2013) compared to Area_sec (˜ 46 % in 2013). The very intense convective events are slightly more numerous in Area_sec. In summary, the thunderstorm activity in Area_sec is more variable at different scales of time (annually and daily), in intensity and in location. Area_max combines two favourable effects for thunderstorm development, the convergence associated with the African easterly jet of the Southern Hemisphere (AEJ-S) and a geographic effect due to the orography and the presence of a lake. The location of the strong convection in Area_sec is modulated by the distance of westward propagation/regeneration of mesoscale convective systems (MCSs) in relation to the phase of Kelvin waves.

The type IX secretion system is required for virulence of the fish pathogen Flavobacterium columnare

USDA-ARS?s Scientific Manuscript database

Flavobacterium columnare, a member of the phylum Bacteroidetes, causes columnaris disease in wild and aquaculture-reared freshwater fish. The mechanisms responsible for columnaris disease are not known. Many members of the phylum Bacteroidetes use type IX secretion systems (T9SSs) to secrete enzymes...

77 FR 53906 - Notice of Proposed Withdrawal and Opportunity for Public Meeting; California

Federal Register 2010, 2011, 2012, 2013, 2014

2012-09-04

...\\; Sec. 32, lots 4 and 5; Sec. 34, lot 4. T. 13 N., R. 10 E., Sec. 2, lot 1, and lots 3 to 15, inclusive; Sec. 9, lots 8, 12, and 13, and SW\\1/4\\NE\\1/4\\; Sec. 10, lots 1 to 10, inclusive, E\\1/2\\NE\\1/4\\, E\\1/2..., inclusive, S\\1/2\\ of lot 5, S\\1/2\\ of lot 8, lots 11 and 13; Sec. 19, lot 24; Sec. 20, lots 1, 2, 3, and 8...

Quantum secret sharing with qudit graph states

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

Keet, Adrian; Fortescue, Ben; Sanders, Barry C.

We present a unified formalism for threshold quantum secret sharing using graph states of systems with prime dimension. We construct protocols for three varieties of secret sharing: with classical and quantum secrets shared between parties over both classical and quantum channels.

Intrinsic and chemically produced microheterogeneity of Staphylococcus aureus enterotoxin type C.

PubMed Central

Metzger, J F; Johnson, A D; Spero, L

1975-01-01

Staphylococcus aureus enterotoxins C1 (SEC1) and C2 (SEC2) produced from 50-liter quantities of crude culture supernatants were purified chromatographically in a neutral or acid milieu. Microheterogenity of SEC1 was markedly increased by treatment of the purified toxin with alkali, and new more acidic charged species appeared. SEC2 was more heterogenous than any of the other S. aureus enterotoxins and was affected only slightly by treatment with alkali. Prolonged incubation of the organism during production of the SEC2 produced changes in charged species that may be related to a bacterial deamidase, since similar changes were not seen with alkaline treatment of the purified toxin. Although SEC1 and SEC2 showed complete identity immunologically, they are separate, distinct toxins, and alkali treatment of SEC1 did not produce SEC2. Images PMID:237837

Pressure scaled water impact test of a 12.5 inch diameter model of the Space Shuttle solid rocket booster

NASA Technical Reports Server (NTRS)

1982-01-01

A total of 59 tail first drops were made. Model entry conditions simulated full scale vertical velocities of approximately 75 to 110 ft/sec with horizontal velocities up to 45 ft/sec and impact angles to + or - 10 deg. These tests were conducted at scaled atmospheric pressures (1.26 psia or 65 mm.Hg). The model, test program, test facility, test equipment, instrumentation system, data reduction procedures, and test results are described.

The development of an interim generalized gate logic software simulator

NASA Technical Reports Server (NTRS)

Mcgough, J. G.; Nemeroff, S.

1985-01-01

A proof-of-concept computer program called IGGLOSS (Interim Generalized Gate Logic Software Simulator) was developed and is discussed. The simulator engine was designed to perform stochastic estimation of self test coverage (fault-detection latency times) of digital computers or systems. A major attribute of the IGGLOSS is its high-speed simulation: 9.5 x 1,000,000 gates/cpu sec for nonfaulted circuits and 4.4 x 1,000,000 gates/cpu sec for faulted circuits on a VAX 11/780 host computer.

75 FR 2197 - Western Pacific Fisheries; Regulatory Restructuring

Federal Register 2010, 2011, 2012, 2013, 2014

2010-01-14

....16 Vessel identification. Sec. 665.17 Experimental fishing..... Sec. 665.17 Experimental fishing. Sec.... 0586, and -0589. Sec. 665.17 Experimental Sec. 665.17 0648-0214 and -0490. fishing. Experimental...) experimental fishing reports estimated at 4 hours (hr) per reporting action; (d) sales and transshipment...

The mechanism of protein export enhancement by the SecDF membrane component

PubMed Central

Tsukazaki, Tomoya; Nureki, Osamu

2011-01-01

Protein transport across membranes is a fundamental and essential cellular activity in all organisms. In bacteria, protein export across the cytoplasmic membrane, driven by dynamic interplays between the protein-conducting SecYEG channel (Sec translocon) and the SecA ATPase, is enhanced by the proton motive force (PMF) and a membrane-integrated Sec component, SecDF. However, the structure and function of SecDF have remained unclear. We solved the first crystal structure of SecDF, consisting of a pseudo-symmetrical 12-helix transmembrane domain and two protruding periplasmic domains. Based on the structural features, we proposed that SecDF functions as a membrane-integrated chaperone, which drives protein movement without using the major energetic currency, ATP, but with remarkable cycles of conformational changes, powered by the proton gradient across the membrane. By a series of biochemical and biophysical approaches, several functionally important residues in the transmembrane region have been identified and our model of the SecDF function has been verified. PMID:27857601

NextGen Flight Deck Surface Trajectory-Based Operations (STBO): Contingency Holds

NASA Technical Reports Server (NTRS)

Bakowski, Deborah Lee; Hooey, Becky Lee; Foyle, David C.; Wolter, Cynthia A.; Cheng, Lara W. S.

2013-01-01

The purpose of this pilot-in-the-loop taxi simulation was to investigate a NextGen Surface Trajectory-Based Operations (STBO) concept called "contingency holds." The contingency-hold concept parses a taxi route into segments, allowing an air traffic control (ATC) surface traffic management (STM) system to hold an aircraft when necessary for safety. Under nominal conditions, if the intersection or active runway crossing is clear, the hold is removed, allowing the aircraft to continue taxiing without slowing, thus improving taxi efficiency, while minimizing the excessive brake use, fuel burn, and emissions associated with stop-and-go taxi. However, when a potential traffic conflict exists, the hold remains in place as a fail-safe mechanism. In this departure operations simulation, the taxi clearance included a required time of arrival (RTA) to a specified intersection. The flight deck was equipped with speed-guidance avionics to aid the pilot in safely meeting the RTA. On two trials, the contingency hold was not released, and pilots were required to stop. On two trials the contingency hold was released 15 sec prior to the RTA, and on two trials the contingency hold was released 30 sec prior to the RTA. When the hold remained in place, all pilots complied with the hold. Results also showed that when the hold was released at 15-sec or 30-sec prior to the RTA, the 30-sec release allowed pilots to maintain nominal taxi speed, thus supporting continuous traffic flow; whereas, the 15-sec release did not. The contingency-hold concept, with at least a 30-sec release, allows pilots to improve taxiing efficiency by reducing braking, slowing, and stopping, but still maintains safety in that no pilots "busted" the clearance holds. Overall, the evidence suggests that the contingency-hold concept is a viable concept for optimizing efficiency while maintaining safety.

A Study of relationship between frailty and physical performance in elderly women.

PubMed

Jeoung, Bog Ja; Lee, Yang Chool

2015-08-01

Frailty is a disorder of multiple inter-related physiological systems. It is unclear whether the level of physical performance factors can serve as markers of frailty and a sign. The purpose of this study was to examine the relationship between frailty and physical performance in elderly women. One hundred fourteen elderly women participated in this study, their aged was from 65 to 80. We were measured 6-min walk test, grip-strength, 30-sec arm curl test, 30-sec chair stand test, 8 foot Up- and Go, Back scratch, chair sit and reach, unipedal stance, BMI, and the frailty with questionnaire. The collected data were analyzed by descriptive statistics, frequencies, correlation analysis, ANOVA, and simple liner regression using the IBM 21. SPSS program. In results, statistic tests showed that there were significant differences between frailty and 6-min walk test, 30-sec arm curl test, 30-sec chair stand test, grip-strength, Back scratch, and BMI. However, we did not find significant differences between frailty and 8 foot Up- and Go, unipedal stance. When the subjects were divided into five groups according to physical performance level, subjects with high 6-min walk, 30-sec arm curl test, chair sit and reach test, and high grip strength had low score frailty. Physical performance factors were strongly associated with decreased frailty, suggesting that physical performance improvements play an important role in preventing or reducing the frailty.

Metabolism of the insecticidally active GABA sub A receptor antagonist 4-sec-(3,4- sup 3 H sub 2 )butyl-1-(4-cyanophenyl)-2,6,7-trioxabicyclo(2. 2. 2)octane

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

Deng, Yanli; Palmer, C.J.; Toia, R.F.

1990-03-01

4-sec-(3,4-{sup 3}H{sub 2})Butyl-1-(4-cyanophenyl)-2,6,7-trioxabicyclo(2.2.2)octane (referred to as ({sup 3}H)COB) was examined as an example of a new class of insecticidally active compounds that block the {gamma}-aminobutyric acid gated chloride channel. Metabolites were identified by thin-layer cochromatography with standards from synthesis and by consideration of their hydrolytic and oxidative degradation products formed in situ on two-dimensional silica gel chromatoplates. Metabolism of ({sup 3}H)COB by mouse liver and housefly abdomen microsomes is dependent on fortification with NADPH. The O-methylene and sec-butyl sites are sensitive to oxidation. Each carbon of the sec-butyl group is individually functionalized with strong preference for the methylene site inmore » the mouse but not the housefly microsomal system. O-Methylene hydroxylation initiates spontaneous cage opening to form an aldehyde that undergoes metabolic reduction, ultimately yielding the same cyanobenzoate ester of 2,2-bis-(hydroxymethyl)-3-methylpentan-1-ol formed by direct hydrolysis. Houseflies injected with ({sup 3}H)COB form many if not all of the same metabolites, with major products being the aforementioned cyanobenzoate, the orthoester oxidized at the sec-butyl methylene site, and polar conjugates.« less

On charge exchange and knock-on processes in the exosphere of Io

NASA Technical Reports Server (NTRS)

Ip, W.-H.

1982-01-01

One direct consequence of magnetospheric interaction of Io is the strong dynamical coupling of its neutral atmosphere with the corotating plasma. The absorption of the thermal ions and the associated neutral injection is an improtant issue not yet explored. As far as nonthermal escape of the neutral atmosphere is concerned, three processes stand out. That is, apart from sputtering, exospheric interactions like atom-ion knock-on collision and charge exchange recombination could be a significant source of the neutral clouds in the Jovian system. Using a current electrodynamic model of Io, both the absorption rate of the corotating thermal plasma and the production rates of new exospheric ions and the fast neutrals are considered. It is found that the source strength of the neutral atoms and molecules with speeds of about 100 km/sec could amount to 10 to the 26th/sec whereas exospheric neutrals emitted at lower speed (of about 10 km/sec) amounts to 4 x 10 to the 25th/sec. The generation of the new ions in connection with the streaming of the magnetospheric plasma around Io could also produce an asymmetric sputtering with a neutral flux of about 10 to the 27th/sec emitted from the region of Io which faces Jupiter. These results may be related to a number of sodium observations.

HST FOS spectroscopy of M87: Evidence for a disk of ionized gas around a massive black hole

NASA Technical Reports Server (NTRS)

Harms, Richard J.; Ford, Holland C.; Tsvetanov, Zlatan I.; Hartig, George F.; Dressel, Linda L.; Kriss, Gerard A.; Bohlin, Ralph; Davidsen, Arthur F.; Margon, Bruce; Kochhar, Ajay K.

1994-01-01

Using the Faint Object Spectrograph (FOS) on the Hubble Space Telescope (HST) to observe the central region of M87, we have obtained spectra covering approximately 4600-6800 A at a spectral dispersion approximately 4.4 A per resolution element through the .26 sec diameter entrance aperture. One spectrum was obtained centered on the nucleus of M87 and two centered 0.25 sec off the nucleus at position angles of 21 deg and 201 deg, thus sampling the anticipated major axis of the disklike structure (described in a companion Letter) expected to lie approximately perpendicular to the axis of the M87 jet. Pointing errors for these observations are estimated to be less than 0.02 sec. Radial velocities of the ionized gas in the two positions 0.25 sec on either side of the nucleus are measured to be approx. equals +/- 500 km/s relative to the M87 systemic velocity. These observations plus emission-line spectra obtained at two additional locations near the nucleus show the ionized gas to be in Keplerian rotation about a mass M = (2.4 +/- 0.7) x 10(exp 9) solar mass within the inner 0.25 sec of M87. Our results provide strong evidence for the presence of a supermassive nuclear black hole in M87.

Normative behavioral thresholds for short tone-bursts.

PubMed

Beattie, R C; Rochverger, I

2001-10-01

Although tone-bursts have been commonly used in auditory brainstem response (ABR) evaluations for many years, national standards describing normal calibration values have not been established. This study was designed to gather normative threshold data to establish a physical reference for tone-burst stimuli that can be reproduced across clinics and laboratories. More specifically, we obtained norms for 3-msec tone-bursts presented at two repetition rates (9.3/sec and 39/sec), two gating functions (Trapezoid and Blackman), and four frequencies (500, 1000, 2000, and 4000 Hz). Our results are specified using three physical references: dB peak sound pressure level, dB peak-to-peak equivalent sound pressure level, and dB SPL (fast meter response, rate = 50 stimuli/sec). These data are offered for consideration when calibrating ABR equipment. The 39/sec stimulus rate yielded tone-burst thresholds that were approximately 3 dB lower than the 9.3/sec rate. The improvement in threshold with increasing stimulus rate may reflect the ability of the auditory system to integrate energy that occurs within a time interval of 200 to 500 msec (temporal integration). The Trapezoid gating function yielded thresholds that averaged 1.4 dB lower than the Blackman function. Although these differences are small and of little clinical importance, the cumulative effects of several instrument and/or procedural variables may yield clinically important differences.

Structure of a PE-PPE-EspG complex from Mycobacterium tuberculosis reveals molecular specificity of ESX protein secretion

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

Ekiert, Damian C.; Cox, Jeffery S.

Nearly 10% of the coding capacity of the Mycobacterium tuberculosis genome is devoted to two highly expanded and enigmatic protein families called PE and PPE, some of which are important virulence/immunogenicity factors and are secreted during infection via a unique alternative secretory system termed "type VII." How PE-PPE proteins function during infection and how they are translocated to the bacterial surface through the five distinct type VII secretion systems [ESAT-6 secretion system (ESX)] of M. tuberculosis is poorly understood. Here in this paper, we report the crystal structure of a PE-PPE heterodimer bound to ESX secretion-associated protein G (EspG), whichmore » adopts a novel fold. This PE-PPE-EspG complex, along with structures of two additional EspGs, suggests that EspG acts as an adaptor that recognizes specific PE-PPE protein complexes via extensive interactions with PPE domains, and delivers them to ESX machinery for secretion. Surprisingly, secretion of most PE-PPE proteins in M. tuberculosis is likely mediated by EspG from the ESX-5 system, underscoring the importance of ESX-5 in mycobacterial pathogenesis. Furthermore, our results indicate that PE-PPE domains function as cis-acting targeting sequences that are read out by EspGs, revealing the molecular specificity for secretion through distinct ESX pathways.« less

Structure of a PE-PPE-EspG complex from Mycobacterium tuberculosis reveals molecular specificity of ESX protein secretion

DOE PAGES

Ekiert, Damian C.; Cox, Jeffery S.

2014-10-01

Nearly 10% of the coding capacity of the Mycobacterium tuberculosis genome is devoted to two highly expanded and enigmatic protein families called PE and PPE, some of which are important virulence/immunogenicity factors and are secreted during infection via a unique alternative secretory system termed "type VII." How PE-PPE proteins function during infection and how they are translocated to the bacterial surface through the five distinct type VII secretion systems [ESAT-6 secretion system (ESX)] of M. tuberculosis is poorly understood. Here in this paper, we report the crystal structure of a PE-PPE heterodimer bound to ESX secretion-associated protein G (EspG), whichmore » adopts a novel fold. This PE-PPE-EspG complex, along with structures of two additional EspGs, suggests that EspG acts as an adaptor that recognizes specific PE-PPE protein complexes via extensive interactions with PPE domains, and delivers them to ESX machinery for secretion. Surprisingly, secretion of most PE-PPE proteins in M. tuberculosis is likely mediated by EspG from the ESX-5 system, underscoring the importance of ESX-5 in mycobacterial pathogenesis. Furthermore, our results indicate that PE-PPE domains function as cis-acting targeting sequences that are read out by EspGs, revealing the molecular specificity for secretion through distinct ESX pathways.« less

Exocyst SEC3 and Phosphoinositides Define Sites of Exocytosis in Pollen Tube Initiation and Growth1[OPEN

PubMed Central

Bloch, Daria; Pleskot, Roman; Vukašinović, Nemanja

2016-01-01

Polarized exocytosis is critical for pollen tube growth, but its localization and function are still under debate. The exocyst vesicle-tethering complex functions in polarized exocytosis. Here, we show that a sec3a exocyst subunit null mutant cannot be transmitted through the male gametophyte due to a defect in pollen tube growth. The green fluorescent protein (GFP)-SEC3a fusion protein is functional and accumulates at or proximal to the pollen tube tip plasma membrane. Partial complementation of sec3a resulted in the development of pollen with multiple tips, indicating that SEC3 is required to determine the site of pollen germination pore formation. Time-lapse imaging demonstrated that SEC3a and SEC8 were highly dynamic and that SEC3a localization on the apical plasma membrane predicts the direction of growth. At the tip, polar SEC3a domains coincided with cell wall deposition. Labeling of GFP-SEC3a-expressing pollen with the endocytic marker FM4-64 revealed the presence of subdomains on the apical membrane characterized by extensive exocytosis. In steady-state growing tobacco (Nicotiana tabacum) pollen tubes, SEC3a displayed amino-terminal Pleckstrin homology-like domain (SEC3a-N)-dependent subapical membrane localization. In agreement, SEC3a-N interacted with phosphoinositides in vitro and colocalized with a phosphatidylinositol 4,5-bisphosphate (PIP2) marker in pollen tubes. Correspondingly, molecular dynamics simulations indicated that SEC3a-N associates with the membrane by interacting with PIP2. However, the interaction with PIP2 is not required for polar localization and the function of SEC3a in Arabidopsis (Arabidopsis thaliana). Taken together, our findings indicate that SEC3a is a critical determinant of polar exocytosis during tip growth and suggest differential regulation of the exocytotic machinery depending on pollen tube growth modes. PMID:27516531

Sec34p, a Protein Required for Vesicle Tethering to the Yeast Golgi Apparatus, Is in a Complex with Sec35p

PubMed Central

VanRheenen, Susan M.; Cao, Xiaochun; Sapperstein, Stephanie K.; Chiang, Elbert C.; Lupashin, Vladimir V.; Barlowe, Charles; Waters, M. Gerard

1999-01-01

A screen for mutants of Saccharomyces cerevisiae secretory pathway components previously yielded sec34, a mutant that accumulates numerous vesicles and fails to transport proteins from the ER to the Golgi complex at the restrictive temperature (Wuestehube, L.J., R. Duden, A. Eun, S. Hamamoto, P. Korn, R. Ram, and R. Schekman. 1996. Genetics. 142:393–406). We find that SEC34 encodes a novel protein of 93-kD, peripherally associated with membranes. The temperature-sensitive phenotype of sec34-2 is suppressed by the rab GTPase Ypt1p that functions early in the secretory pathway, or by the dominant form of the ER to Golgi complex target-SNARE (soluble N-ethylmaleimide sensitive fusion protein attachment protein receptor)–associated protein Sly1p, Sly1-20p. Weaker suppression is evident upon overexpression of genes encoding the vesicle tethering factor Uso1p or the vesicle-SNAREs Sec22p, Bet1p, or Ykt6p. This genetic suppression profile is similar to that of sec35-1, a mutant allele of a gene encoding an ER to Golgi vesicle tethering factor and, like Sec35p, Sec34p is required in vitro for vesicle tethering. sec34-2 and sec35-1 display a synthetic lethal interaction, a genetic result explained by the finding that Sec34p and Sec35p can interact by two-hybrid analysis. Fractionation of yeast cytosol indicates that Sec34p and Sec35p exist in an ∼750-kD protein complex. Finally, we describe RUD3, a novel gene identified through a genetic screen for multicopy suppressors of a mutation in USO1, which suppresses the sec34-2 mutation as well. PMID:10562277

Exocyst SEC3 and Phosphoinositides Define Sites of Exocytosis in Pollen Tube Initiation and Growth.

PubMed

Bloch, Daria; Pleskot, Roman; Pejchar, Přemysl; Potocký, Martin; Trpkošová, Pavlína; Cwiklik, Lukasz; Vukašinović, Nemanja; Sternberg, Hasana; Yalovsky, Shaul; Žárský, Viktor

2016-10-01

Polarized exocytosis is critical for pollen tube growth, but its localization and function are still under debate. The exocyst vesicle-tethering complex functions in polarized exocytosis. Here, we show that a sec3a exocyst subunit null mutant cannot be transmitted through the male gametophyte due to a defect in pollen tube growth. The green fluorescent protein (GFP)-SEC3a fusion protein is functional and accumulates at or proximal to the pollen tube tip plasma membrane. Partial complementation of sec3a resulted in the development of pollen with multiple tips, indicating that SEC3 is required to determine the site of pollen germination pore formation. Time-lapse imaging demonstrated that SEC3a and SEC8 were highly dynamic and that SEC3a localization on the apical plasma membrane predicts the direction of growth. At the tip, polar SEC3a domains coincided with cell wall deposition. Labeling of GFP-SEC3a-expressing pollen with the endocytic marker FM4-64 revealed the presence of subdomains on the apical membrane characterized by extensive exocytosis. In steady-state growing tobacco (Nicotiana tabacum) pollen tubes, SEC3a displayed amino-terminal Pleckstrin homology-like domain (SEC3a-N)-dependent subapical membrane localization. In agreement, SEC3a-N interacted with phosphoinositides in vitro and colocalized with a phosphatidylinositol 4,5-bisphosphate (PIP 2 ) marker in pollen tubes. Correspondingly, molecular dynamics simulations indicated that SEC3a-N associates with the membrane by interacting with PIP 2 However, the interaction with PIP 2 is not required for polar localization and the function of SEC3a in Arabidopsis (Arabidopsis thaliana). Taken together, our findings indicate that SEC3a is a critical determinant of polar exocytosis during tip growth and suggest differential regulation of the exocytotic machinery depending on pollen tube growth modes. © 2016 American Society of Plant Biologists. All Rights Reserved.

78 FR 73466 - Privacy Act

Federal Register 2010, 2011, 2012, 2013, 2014

2013-12-06

... Subpart A--General. Sec. 707.11 Scope and purpose. Sec. 707.12 Definitions. Sec. 707.13 Preservation of records. Subpart B--Requests for access to records; amendment of records, accounting of disclosures.... Sec. 707.23 Requests for amendment of records. Sec. 707.24 Requests for an accounting of record...

32 CFR 2001.46 - Transmission.

Code of Federal Regulations, 2010 CFR

2010-07-01

... between the U.S., Puerto Rico, or a U.S. possession or trust territory. (1) Top Secret. Top Secret... an authorized government agency courier service; a designated courier or escort with Top Secret clearance; electronic means over approved communications systems. Under no circumstances will Top Secret...

Role of solid-phase microextraction in the identification of highly volatile pheromones of two Rhinoceros beetles Scapanes australis and Strategus aloeus (Coleoptera, Scarabaeidae, Dynastinae).

PubMed

Rochat, D; Ramirez-Lucas, P; Malosse, C; Aldana, R; Kakul, T; Morin, J P

2000-07-14

Solid-phase microextraction (SPME) samplings from live insects or natural secretion allowed one to identify the aggregation pheromones of the pest beetles Scapanes australis and Strategus aloeus by efficient and rapid isolation of their highly volatile (72 < M(r) < 116) components. S. australis male pheromone was identified as a 84:12:4 (w/w) mixture of 2-butanol [67:33 (R)-(-):(S)-(+) ratio], 3-hydroxy-2-butanone and 2,3-butanediol [43:17:40 (R,R)-(-):(S,S)-(+):meso ratio], and S. aloeus pheromone as a 95.5:4.0:0.5 (w/w) mixture of 2-butanone, 3-pentanone and sec.-butyl acetate by GC-MS using conventional and chiral capillary columns. This is the first report of Scarabaeidae pheromones based on such small and common molecules.

[Influence of honey, royal jelly and propolis on accelerating acetate healing of experimental gastric ulcers in rats].

PubMed

Belostotskiĭ, N I; Kas'ianenko, V I; Dubtsova, E A; Lazebnik, L B

2009-01-01

This study examines gastric acetic ulcer healing in the rat after administration of honey, royal jelly and propolis into the stomach. Chronic gastric ulcers were induced in male Wistar rats by the application of 100% acetic acid to the serosal surface of the stomach on 60 sec. Bee-keeping products were administrated into the stomach from 2nd to 7th day after acetic ulcer induction. On 7th day animals were killed, and ulcer area was measured in mm2. In gastric juice pH and activity of pepsin were measured. The healing of acetic ulcers is accelerated with the administration of honey, royal jelly or propolis during six days. The largest healing effect was demonstrated with propolis and royal jelly, smaller one with the honey. It was revealed decrease of stomach acid secretion in the rats, which have received bee-keeping products versus the rats of control group.

BEARS: Radioactive ion beams at LBNL

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

Powell, J.; Guo, F.Q.; Haustein, P.E.

1998-07-01

BEARS (Berkeley Experiments with Accelerated Radioactive Species) is an initiative to develop a radioactive ion-beam capability at Lawrence Berkeley National Laboratory. The aim is to produce isotopes at an existing medical cyclotron and to accelerate them at the 88 inch Cyclotron. To overcome the 300-meter physical separation of these two accelerators, a carrier-gas transport system will be used. At the terminus of the capillary, the carrier gas will be separated and the isotopes will be injected into the 88 inch Cyclotron`s Electron Cyclotron Resonance (ECR) ion source. The first radioactive beams to be developed will include 20-min {sup 11}C andmore » 70-sec {sup 14}O, produced by (p,n) and (p,{alpha}) reactions on low-Z targets. A test program is currently being conducted at the 88 inch Cyclotron to develop the parts of the BEARS system. Preliminary results of these tests lead to projections of initial {sup 11}C beams of up to 2.5 {times} 10{sup 7} ions/sec and {sup 14}O beams of 3 {times} 10{sup 5} ions/sec.« less

Monitoring substrate enables real-time regulation of a protein localization pathway.

PubMed

Ito, Koreaki; Mori, Hiroyuki; Chiba, Shinobu

2018-06-01

Protein localization machinery supports cell survival and physiology, suggesting the potential importance of its expression regulation. Here, we summarize a remarkable scheme of regulation, which allows real-time feedback regulation of the machinery expression. A class of regulatory nascent polypeptides, called monitoring substrates, undergoes force-sensitive translation arrest. The resulting ribosome stalling on the mRNA then affects mRNA folding to expose the ribosome-binding site of the downstream target gene and upregulate its translation. The target gene encodes a component of the localization machinery, whose physical action against the monitoring substrate leads to arrest cancellation. Thus, this scheme of feedback loop allows the cell to adjust the amount of the machinery to correlate inversely with the effectiveness of the process at a given moment. The system appears to have emerged late in evolution, in which a narrow range of organisms selected a distinct monitoring substrate-machinery combination. Currently, regulatory systems of SecM-SecA, VemP-SecDF2 and MifM-YidC2 are known to occur in different bacterial species.

Channel crossing: how are proteins shipped across the bacterial plasma membrane?

PubMed

Collinson, Ian; Corey, Robin A; Allen, William J

2015-10-05

The structure of the first protein-conducting channel was determined more than a decade ago. Today, we are still puzzled by the outstanding problem of protein translocation--the dynamic mechanism underlying the consignment of proteins across and into membranes. This review is an attempt to summarize and understand the energy transducing capabilities of protein-translocating machines, with emphasis on bacterial systems: how polypeptides make headway against the lipid bilayer and how the process is coupled to the free energy associated with ATP hydrolysis and the transmembrane protein motive force. In order to explore how cargo is driven across the membrane, the known structures of the protein-translocation machines are set out against the background of the historic literature, and in the light of experiments conducted in their wake. The paper will focus on the bacterial general secretory (Sec) pathway (SecY-complex), and its eukaryotic counterpart (Sec61-complex), which ferry proteins across the membrane in an unfolded state, as well as the unrelated Tat system that assembles bespoke channels for the export of folded proteins. © 2015 The Authors.

A translocator-specific export signal establishes the translocator-effector secretion hierarchy that is important for type III secretion system function

PubMed Central

Tomalka, Amanda G.; Stopford, Charles M.; Lee, Pei-Chung; Rietsch, Arne

2012-01-01

Summary Type III secretion systems are used by many Gram-negative pathogens to directly deliver effector proteins into the cytoplasm of host cells. To accomplish this, bacteria secrete translocator proteins that form a pore in the host-cell membrane through which the effector proteins are then introduced into the host cell. Evidence from multiple systems indicates that the pore-forming translocator proteins are exported before effectors, but how this secretion hierarchy is established is unclear. Here we used the P. aeruginosa translocator protein PopD as a model to identify its export signals. The amino-terminal secretion signal and chaperone, PcrH, are required for export under all conditions. Two novel signals in PopD, one proximal to the chaperone-binding site and one at the very C-terminus of the protein, are required for export of PopD before effector proteins. These novel export signals establish the translocator-effector secretion hierarchy, which in turn, is critical for the delivery of effectors into host cells. PMID:23121689

75 FR 74743 - Notice of Proposed Withdrawal Extension, Corrections to Existing Withdrawal, and Opportunity for...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-12-01

.... 11 to 14, inclusive; Sec. 15, NE\\1/4\\, W\\1/2\\NW\\1/4\\, NW\\1/4\\SW\\1/4\\, and E\\1/2\\SE\\1/ 4\\; Sec. 19...\\; Secs. 23 to 34, inclusive; Sec. 35, NE\\1/4\\, NW\\1/4\\NW\\1/4\\, S\\1/2\\NW\\1/4\\, and S\\1/2\\; Sec. 36. T. 46 N., R. 22 E., Secs. 1 to 5, inclusive; Sec. 6, lots 1 and 2, S\\1/2\\NE\\1/4\\, E\\1/2\\SW\\1/4\\, and SE\\1...

A rule-based approach for the correlation of alarms to support Disaster and Emergency Management

NASA Astrophysics Data System (ADS)

Gloria, M.; Minei, G.; Lersi, V.; Pasquariello, D.; Monti, C.; Saitto, A.

2009-04-01

Key words: Simple Event Correlator, Agent Platform, Ontology, Semantic Web, Distributed Systems, Emergency Management The importance of recognition of emergency's typology to control the critical situation for security of citizens has been always recognized. It follows this aspect is very important for proper management of a hazardous event. In this work we present a solution for the recognition of emergency's typology adopted by an Italian research project, called CI6 (Centro Integrato per Servizi di Emergenza Innovativi). In our approach, CI6 receives alarms by citizen or people involved in the work (for example: police, operator of 112, and so on). CI6 represents any alarm by a set of information, including a text that describes it and obtained when the user points out the danger, and a pair of coordinates for its location. The system realizes an analysis of text and automatically infers information on the type of emergencies by means a set of parsing rules and rules of inference applied by a independent module: a correlator of events based on their log and called Simple Event Correlator (SEC). SEC, integrated in CI6's platform, is an open source and platform independent event correlation tool. SEC accepts input both files and text derived from standard input, making it flexible because it can be matched to any application that is able to write its output to a file stream. The SEC configuration is stored in text files as rules, each rule specifying an event matching condition, an action list, and optionally a Boolean expression whose truth value decides whether the rule can be applied at a given moment. SEC can produce output events by executing user-specified shell scripts or programs, by writing messages to files, and by various other means. SEC has been successfully applied in various domains like network management, system monitoring, data security, intrusion detection, log file monitoring and analysis, etc; it has been used or integrated with many application as CiscoWorks, HP OpenView NNM and Operation, BMC Patrol, etc. Analysis of text of an alarm can detect some keywords that allow to classify the particular event. The inference rules were developed by means an analysis about news regard real emergency found by web reaserches. We have seen that often a kind of emergency is characterized by more keyword. Keywords are not uniquely associated with a specific emergency, but they can be shared by different types of emergencies (such as. keyword "landslide" can be associated both emergency "landslide" and emergency "Flood"). However, the identification of two or more keywords associated with a particular type of emergency identified in most cases the correct type of emergency. So, for example, if text contains words as "water", "flood", "overflowing", "landslide" o other words belonging to the set of defined keywords or words that have some root of keywords, the system "decides" that this alarm belongs to specific typology, in this case "flood typology". The system has the memory of this information, so if a new alarm is reported and belongs to one of the typology already identified, it proceeds with the comparison of coordinates. The comparison between the centers of the alarms allows to see if they describe an area inscribed in an ideal circle that has centered on the first alarm and radius defined by the typology above mentioned. If this happens the system CI6 creates an emergency that has centered on the centre of that area and typology equal to that of the alarms. It follows that an emergency is represented by at least two alarms. Thus, the system suggests to manager (CI6's user) the possibility that most alarms can concern same events and makes a classification of this event. It is important to stress that CI6 is a system of decision support, hence also this service is limited to providing advice to the user to facilitate his task, leaving him the decision to accept it or not. REFERENCES SEC (Simple Event Correlator), http://kodu.neti.ee/~risto/sec/ M. Gloria,V. Lersi, G. Minei, D. Pasquariello, C. Monti, A. Saitto, "A Semantic WEB Services Platform to support Disaster and Emergency Management", 4th biennial Meeting of International Environmental Modelling and Software Society (iEMSs), 2008

Mathematical Modelling of Drying Kinetics of Wheat in Electron Fired Fluidized Bed Drying System

NASA Astrophysics Data System (ADS)

Deomore, Dayanand N.; Yarasu, Ravindra B.

2018-02-01

The conventional method of electrical heating is replaced by electron firing system. The drying kinetics of wheat is studied using electron fired fluidized bed dryer. The results are simulated by using ANSYS. It was observed that the graphs are in agreement with each other. Therefore, the new proposed electronic firing system can be employed instead of electrical firing. It was observed that the drop in Relative Humidity in case of Electrical heating is 68.75% for temp reaching up to 70° C in 67 sec for pressure drop of 13 psi while for the electronic Firing system it is 67.6 % temp reaches to 70° C in 70 sec for pressure drop of 12.67 psi. As the results are in agreement with each other it was concluded that for the grains like wheat which has low initial moisture content both systems can be used.

Accurate prediction of secreted substrates and identification of a conserved putative secretion signal for type III secretion systems

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

Samudrala, Ram; Heffron, Fred; McDermott, Jason E.

2009-04-24

The type III secretion system is an essential component for virulence in many Gram-negative bacteria. Though components of the secretion system apparatus are conserved, its substrates, effector proteins, are not. We have used a machine learning approach to identify new secreted effectors. The method integrates evolutionary measures, such as the pattern of homologs in a range of other organisms, and sequence-based features, such as G+C content, amino acid composition and the N-terminal 30 residues of the protein sequence. The method was trained on known effectors from Salmonella typhimurium and validated on a corresponding set of effectors from Pseudomonas syringae, aftermore » eliminating effectors with detectable sequence similarity. The method was able to identify all of the known effectors in P. syringae with a specificity of 84% and sensitivity of 82%. The reciprocal validation, training on P. syringae and validating on S. typhimurium, gave similar results with a specificity of 86% when the sensitivity level was 87%. These results show that type III effectors in disparate organisms share common features. We found that maximal performance is attained by including an N-terminal sequence of only 30 residues, which agrees with previous studies indicating that this region contains the secretion signal. We then used the method to define the most important residues in this putative secretion signal. Finally, we present novel predictions of secreted effectors in S. typhimurium, some of which have been experimentally validated, and apply the method to predict secreted effectors in the genetically intractable human pathogen Chlamydia trachomatis. This approach is a novel and effective way to identify secreted effectors in a broad range of pathogenic bacteria for further experimental characterization and provides insight into the nature of the type III secretion signal.« less

Molecular characterization of a functional type VI secretion system from a clinical isolate of Aeromonas hydrophila

EPA Science Inventory

Our laboratory recently molecularly characterized the type II secretion system (T2SS)-associated cytotoxic enterotoxin (Act) and the T3SS-secreted AexU effector from a diarrheal isolate SSU of Aeromonas hydrophila. The role of these toxin proteins in the pathogenesis of A. hydrop...

Molecular Characterization of a Functional Type VI Secretion System from a Clinical Isolate of Aeromonas hydrophilia

EPA Science Inventory

Our laboratory recently molecularly characterized the type II secretion system (T2SS)-associated cytotoxic enterotoxin (Act) and the T3SS-secreted AexU effector from a diarrheal isolate SSU of Aeromonas hydrophila. The role of these toxin proteins in the pathogenesis of A. hydrop...

A protein secretion system linked to bacteroidete gliding motility and pathogenesis

PubMed Central

Sato, Keiko; Naito, Mariko; Yukitake, Hideharu; Hirakawa, Hideki; Shoji, Mikio; McBride, Mark J.; Rhodes, Ryan G.; Nakayama, Koji

2009-01-01

Porphyromonas gingivalis secretes strong proteases called gingipains that are implicated in periodontal pathogenesis. Protein secretion systems common to other Gram-negative bacteria are lacking in P. gingivalis, but several proteins, including PorT, have been linked to gingipain secretion. Comparative genome analysis and genetic experiments revealed 11 additional proteins involved in gingipain secretion. Six of these (PorK, PorL, PorM, PorN, PorW, and Sov) were similar in sequence to Flavobacterium johnsoniae gliding motility proteins, and two others (PorX and PorY) were putative two-component system regulatory proteins. Real-time RT-PCR analysis revealed that porK, porL, porM, porN, porP, porT, and sov were down-regulated in P. gingivalis porX and porY mutants. Disruption of the F. johnsoniae porT ortholog resulted in defects in motility, chitinase secretion, and translocation of a gliding motility protein, SprB adhesin, to the cell surface, providing a link between a unique protein translocation system and a motility apparatus in members of the Bacteroidetes phylum. PMID:19966289

Multi-fault clustering and diagnosis of gear system mined by spectrum entropy clustering based on higher order cumulants

NASA Astrophysics Data System (ADS)

Shao, Renping; Li, Jing; Hu, Wentao; Dong, Feifei

2013-02-01

Higher order cumulants (HOC) is a new kind of modern signal analysis of theory and technology. Spectrum entropy clustering (SEC) is a data mining method of statistics, extracting useful characteristics from a mass of nonlinear and non-stationary data. Following a discussion on the characteristics of HOC theory and SEC method in this paper, the study of signal processing techniques and the unique merits of nonlinear coupling characteristic analysis in processing random and non-stationary signals are introduced. Also, a new clustering analysis and diagnosis method is proposed for detecting multi-damage on gear by introducing the combination of HOC and SEC into the damage-detection and diagnosis of the gear system. The noise is restrained by HOC and by extracting coupling features and separating the characteristic signal at different speeds and frequency bands. Under such circumstances, the weak signal characteristics in the system are emphasized and the characteristic of multi-fault is extracted. Adopting a data-mining method of SEC conducts an analysis and diagnosis at various running states, such as the speed of 300 r/min, 900 r/min, 1200 r/min, and 1500 r/min of the following six signals: no-fault, short crack-fault in tooth root, long crack-fault in tooth root, short crack-fault in pitch circle, long crack-fault in pitch circle, and wear-fault on tooth. Research shows that this combined method of detection and diagnosis can also identify the degree of damage of some faults. On this basis, the virtual instrument of the gear system which detects damage and diagnoses faults is developed by combining with advantages of MATLAB and VC++, employing component object module technology, adopting mixed programming methods, and calling the program transformed from an *.m file under VC++. This software system possesses functions of collecting and introducing vibration signals of gear, analyzing and processing signals, extracting features, visualizing graphics, detecting and diagnosing faults, detecting and monitoring, etc. Finally, the results of testing and verifying show that the developed system can effectively be used to detect and diagnose faults in an actual operating gear transmission system.

Multi-fault clustering and diagnosis of gear system mined by spectrum entropy clustering based on higher order cumulants.

PubMed

Shao, Renping; Li, Jing; Hu, Wentao; Dong, Feifei

2013-02-01

Higher order cumulants (HOC) is a new kind of modern signal analysis of theory and technology. Spectrum entropy clustering (SEC) is a data mining method of statistics, extracting useful characteristics from a mass of nonlinear and non-stationary data. Following a discussion on the characteristics of HOC theory and SEC method in this paper, the study of signal processing techniques and the unique merits of nonlinear coupling characteristic analysis in processing random and non-stationary signals are introduced. Also, a new clustering analysis and diagnosis method is proposed for detecting multi-damage on gear by introducing the combination of HOC and SEC into the damage-detection and diagnosis of the gear system. The noise is restrained by HOC and by extracting coupling features and separating the characteristic signal at different speeds and frequency bands. Under such circumstances, the weak signal characteristics in the system are emphasized and the characteristic of multi-fault is extracted. Adopting a data-mining method of SEC conducts an analysis and diagnosis at various running states, such as the speed of 300 r/min, 900 r/min, 1200 r/min, and 1500 r/min of the following six signals: no-fault, short crack-fault in tooth root, long crack-fault in tooth root, short crack-fault in pitch circle, long crack-fault in pitch circle, and wear-fault on tooth. Research shows that this combined method of detection and diagnosis can also identify the degree of damage of some faults. On this basis, the virtual instrument of the gear system which detects damage and diagnoses faults is developed by combining with advantages of MATLAB and VC++, employing component object module technology, adopting mixed programming methods, and calling the program transformed from an *.m file under VC++. This software system possesses functions of collecting and introducing vibration signals of gear, analyzing and processing signals, extracting features, visualizing graphics, detecting and diagnosing faults, detecting and monitoring, etc. Finally, the results of testing and verifying show that the developed system can effectively be used to detect and diagnose faults in an actual operating gear transmission system.

Study of magnetic perturbations on SEC vidicon tubes. [large space telescope

NASA Technical Reports Server (NTRS)

Long, D. C.; Zucchino, P.; Lowrance, J.

1973-01-01

A laboratory measurements program was conducted to determine the tolerances that must be imposed to achieve optimum performance from SEC-vidicon data sensors in the LST mission. These measurements along with other data were used to formulate recommendations regarding the necessary telemetry and remote control for the television data sensors when in orbit. The study encompassed the following tasks: (1) Conducted laboratory measurements of the perturbations which an external magnetic field produces on a magnetically focused, SEC-vidicon. Evaluated shielding approaches. (2) Experimentally evaluated the effects produced on overall performance by variations of the tube electrode potentials, and the focus, deflection and alignment fields. (3) Recommended the extent of ground control of camera parameters and camera parameter telemetry required for optimizing the performance of the television system in orbit. The experimental data are summarized in a set of graphs.

Discovery of the type VII ESX-1 secretion needle?

PubMed

Ates, Louis S; Brosch, Roland

2017-01-01

Mycobacterium tuberculosis, the etiological agent of human tuberculosis, harbours five ESAT-6/type VII secretion (ESX/T7S) systems. The first esx gene clusters were identified during the genome-sequencing project of M. tuberculosis H37Rv. Follow-up studies revealed additional genes playing important roles in ESX/T7S systems. Among the latter genes, one can find those that encode Pro-Glu (PE) and Pro-Pro-Glu (PPE) proteins as well as a gene cluster that is encoded >260 kb upstream of the esx-1 locus and encodes ESX-1 secretion-associated proteins EspA (Rv3616c), EspC (Rv3615c) and EspD (Rv3614c). The espACD cluster has been suggested to have an important function in ESX-1 secretion since EspA-EspC and EsxA-EsxB are mutually co-dependent on each other for secretion. However, the molecular mechanism of this co-dependence and interaction between the substrates remained unknown. In this issue of Molecular Microbiology, Lou and colleagues show that EspC forms high-molecular weight polymerization complexes that resemble selected components of type II, III and/or IV secretion systems of Gram-negative bacteria. Indeed, EspC-multimeric complexes form filamentous structures that could well represent a secretion needle of ESX-1 type VII secretion systems. This exciting observation opens new avenues for research to discover and characterize ESX/T7S components and elucidates the co-dependence of EsxA/B secretion with EspA/C. © 2016 John Wiley & Sons Ltd.

14 CFR 29.1309 - Equipment, systems, and installations.

Code of Federal Regulations, 2011 CFR

2011-01-01

... effects of lightning strikes on the rotorcraft must be considered. (Secs. 313(a), 601, 603, 604, and 605... Equipment, systems, and installations. (a) The equipment, systems, and installations whose functioning is... functions under any foreseeable operating condition. (b) The rotorcraft systems and associated components...

14 CFR 29.1309 - Equipment, systems, and installations.

Code of Federal Regulations, 2010 CFR

2010-01-01

... effects of lightning strikes on the rotorcraft must be considered. (Secs. 313(a), 601, 603, 604, and 605... Equipment, systems, and installations. (a) The equipment, systems, and installations whose functioning is... functions under any foreseeable operating condition. (b) The rotorcraft systems and associated components...

Experimental continuous sludge microwave system to enhance dehydration ability and hydrogen production from anaerobic digestion of sludge.

PubMed

Zhou, Cuihong; Huang, Xintong; Zeng, Meng

2018-05-01

Dehydrating large amounts of sludge produced by sewage treatment plants is difficult. Microwave pretreatment can effectively and significantly improve the dewaterability and hydrogen production of sludge subjected to anaerobic digestion. The aim of this study was to investigate the effects of different microwave conditions on hydrogen production from anaerobic digestion and dewaterability of sludge. Based on an analysis of the electric field distribution, a spiral reactor was designed and a continuous microwave system was built to conduct intermittent and continuous experiments under different conditions. Settling Volume, Capillary Suction Time, particle size, and moisture content of the sludge were measured. The results show that sludge pretreatment in continuous experiments has equally remarkable dehydration performance as in intermittent experiments; the minimum moisture content was 77.29% in the intermittent experiment under a microwave power of 300W and an exposure time of 60sec, and that in the continuous experiment was 77.56% under a microwave power of 400W and an exposure time of 60sec. The peak measured by Differential Scanning Calorimeter appeared earliest under a microwave power of 600W and an exposure time of 180sec. The heat flux at the peak was 4.343W/g, which is relatively small. This indicates that microwave pretreatment induced desirable effects. The maximum yield of hydrogen production was 7.967% under the conditions of microwave power of 500W, exposure time of 120sec, and water bath at 55°C. This research provides a theoretical and experimental basis for the development of a continuous microwave sludge-conditioning system. Copyright © 2017. Published by Elsevier B.V.

Computerized Orders with Standardized Concentrations Decrease Dispensing Errors of Continuous Infusion Medications for Pediatrics

PubMed Central

Sowan, Azizeh K.; Vaidya, Vinay U.; Soeken, Karen L.; Hilmas, Elora

2010-01-01

OBJECTIVES The use of continuous infusion medications with individualized concentrations may increase the risk for errors in pediatric patients. The objective of this study was to evaluate the effect of computerized prescriber order entry (CPOE) for continuous infusions with standardized concentrations on frequency of pharmacy processing errors. In addition, time to process handwritten versus computerized infusion orders was evaluated and user satisfaction with CPOE as compared to handwritten orders was measured. METHODS Using a crossover design, 10 pharmacists in the pediatric satellite within a university teaching hospital were given test scenarios of handwritten and CPOE order sheets and asked to process infusion orders using the pharmacy system in order to generate infusion labels. Participants were given three groups of orders: five correct handwritten orders, four handwritten orders written with deliberate errors, and five correct CPOE orders. Label errors were analyzed and time to complete the task was recorded. RESULTS Using CPOE orders, participants required less processing time per infusion order (2 min, 5 sec ± 58 sec) compared with time per infusion order in the first handwritten order sheet group (3 min, 7 sec ± 1 min, 20 sec) and the second handwritten order sheet group (3 min, 26 sec ± 1 min, 8 sec), (p<0.01). CPOE eliminated all error types except wrong concentration. With CPOE, 4% of infusions processed contained errors, compared with 26% of the first group of handwritten orders and 45% of the second group of handwritten orders (p<0.03). Pharmacists were more satisfied with CPOE orders when compared with the handwritten method (p=0.0001). CONCLUSIONS CPOE orders saved pharmacists' time and greatly improved the safety of processing continuous infusions, although not all errors were eliminated. pharmacists were overwhelmingly satisfied with the CPOE orders PMID:22477811

Field testing and adaptation of a methodology to measure "in-stream" values in the Tongue River, northern Great Plains (NGP) region

USGS Publications Warehouse

Bovee, Ken D.; Gore, James A.; Silverman, Arnold J.

1978-01-01

A comprehensive, multi-component in-stream flow methodology was developed and field tested in the Tongue River in southeastern Montana. The methodology incorporates a sensitivity for the flow requirements of a wide variety of in-stream uses, and the flexibility to adjust flows to accommodate seasonal and sub-seasonal changes in the flow requirements for different areas. In addition, the methodology provides the means to accurately determine the magnitude of the water requirement for each in-stream use. The methodology can be a powerful water management tool in that it provides the flexibility and accuracy necessary in water use negotiations and evaluation of trade-offs. In contrast to most traditional methodologies, in-stream flow requirements were determined by additive independent methodologies developed for: 1) fisheries, including spawning, rearing, and food production; 2) sediment transport; 3) the mitigation of adverse impacts of ice; and 4) evapotranspiration losses. Since each flow requirement varied in important throughout the year, the consideration of a single in-stream use as a basis for a flow recommendation is inadequate. The study shows that the base flow requirement for spawning shovelnose sturgeon was 13.0 m3/sec. During the same period of the year, the flow required to initiate the scour of sediment from pools is 18.0 m3/sec, with increased scour efficiency occurring at flows between 20.0 and 25.0 m3/sec. An over-winter flow of 2.83 m3/sec. would result in the loss of approximately 80% of the riffle areas to encroachment by surface ice. At the base flow for insect production, approximately 60% of the riffle area is lost to ice. Serious damage to the channel could be incurred from ice jams during the spring break-up period. A flow of 12.0 m3/sec. is recommended to alleviate this problem. Extensive ice jams would be expected at the base rearing and food production levels. The base rearing flow may be profoundly influenced by the loss of streamflow to transpiration. Transpiration losses to riparian vegetation ranged from 0.78 m3/sec. in April, to 1.54 m3/sec. in July, under drought conditions. Requirement for irrigation were estimated to range from 5.56 m3/sec. in May to 7.97 m3/sec. in July, under drought conditions. It was concluded that flow requirements to satisfy monthly water losses to transpiration must be added to the base fishery flows to provide adequate protection to the resources in the lower reaches of the river. Integration of the in-stream requirements for various use components shows that a base flow of at least 23.6 m3/sec. must be reserved during the month of June to initiate scour of sediment from pools, provide spawning habitat to shovelnose sturgeon, and to accommodate water losses from the system. In comparison, a base flow of 3.85 m3/sec. would be required during early February to provide fish rearing habitat and insect productivity, and to prevent excessive loss of food production areas to surface ice formation. During mid to late February, a flow of 12 m3/sec. would be needed to facilitate ice break-up and prevent ice jams from forming. Following break-up, the base flow would again be 3.85 m3/sec. until the start of spawning season.

Selective Photoaffinity Labeling Identifies the Signal Peptide Binding Domain on SecA

PubMed Central

Musial-Siwek, Monika; Rusch, Sharyn L.; Kendall, Debra A.

2007-01-01

SecA, an ATPase crucial to the Sec-dependent translocation machinery in Escherichia coli, recognizes and directly binds the N-terminal signal peptide of an exported preprotein. This interaction plays a central role in the targeting and transport of preproteins via the SecYEG channel. Here we identify the Signal Peptide Binding Groove (SPBG) on SecA addressing a key issue regarding the SecA-preprotein interaction. We employ a synthetic signal peptide containing the photoreactive benzoylphenylalanine to efficiently and specifically label SecA containing a unique Factor Xa site. Comparison of the photolabeled fragment from the subsequent proteolysis of several SecAs, which vary only in the location of the Factor Xa site, reveals one 53-residue segment in common with the entire series. The covalently modified SecA segment produced is the same in aqueous solution and in lipid vesicles. This spans amino acids 269 to 322 of the E. coli protein, which is distinct from a previously proposed signal peptide binding site, and contributes to a hydrophobic peptide binding groove evident in molecular models of SecA. PMID:17084862

Spark ignited turbulent flame kernel growth. Annual report, January--December 1991

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

Santavicca, D.A.

1994-06-01

An experimental study of the effect of spark power on the growth rate of spark-ignited flame kernels was conducted in a turbulent flow system at 1 atm, 300 K conditions. All measurements were made with premixed, propane-air at a fuel/air equivalence ratio of 0.93, with 0%, 8% or 14% dilution. Two flow conditions were studied: a low turbulence intensity case with a mean velocity of 1.25 m/sec and a turbulence intensity of 0.33 m/sec, and a high turbulence intensity case with a mean velocity of 1.04 m/sec and a turbulence intensity of 0.88 m/sec. The growth of the spark-ignited flamemore » kernel was recorded over a time interval from 83 {mu}sec to 20 msec following the start of ignition using high speed laser shadowgraphy. In order to evaluate the effect of ignition spark power, tests were conducted with a long duration (ca 4 msec) inductive discharge ignition system with an average spark power of ca 14 watts and two short duration (ca 100 nsec) breakdown ignition systems with average spark powers of ca 6 {times} 10{sup 4} and ca 6 {times} 10{sup 5} watts. The results showed that increased spark power resulted in an increased growth rate, where the effect of short duration breakdown sparks was found to persist for times of the order of milliseconds. The effectiveness of increased spark power was found to be less at high turbulence and high dilution conditions. Increased spark power had a greater effect on the 0--5 mm burn time than on the 5--13 mm burn time, in part because of the effect of breakdown energy on the initial size of the flame kernel. And finally, when spark power was increased by shortening the spark duration while keeping the effective energy the same there was a significant increase in the misfire rate, however when the spark power was further increased by increasing the breakdown energy the misfire rate dropped to zero.« less

Investigation of Point Doppler Velocimetry (PDV) for Transition Detection in Boundary Layers

NASA Technical Reports Server (NTRS)

Kuhlman, John M.

1999-01-01

A two component Point Doppler Velocimetry (PDV) system has been developed and tested. Improvements were made to an earlier PDV system, in terms of experimental techniques, as well as the data acquisition and reduction software. Measurements of the streamwise and spanwise mean and fluctuating velocities for flows from a rectangular channel and over an NACA 0012 airfoil were made, and the data were compared against hot wire data. The closest to the airfoil surface that PDV measurements could be made was on the order of 0.005 m(0.2", z/c = 0.0169). When the PDV and hot wire data were compared, the time traces for each appeared similar. The mean velocities agreed to within plus or minus 2 m/sec, while the RMS velocities agreed to plus or minus 0.4 m/sec. While the PDV time autocorrelations agreed with those of the hot wire data, the PDV power spectral densities were noisier above 750 Hz. A major source of error in these experiments was determined to be the drifting of the iodine cell stem temperatures. While the stem temperatures were controlled to within plus or minus 0.1 C, this could lead to a frequency shift of as much as 6 MHz, which translates into an error of 1.6 m/sec for the back scatter channel, and up to 6.9 m/sec for the forward scatter channel. These error estimates are consistent with the observed error magnitudes.

Targeting bacterial secretion systems: benefits of disarmament in the microcosm.

PubMed

Baron, Christian; Coombes, Brian

2007-03-01

Secretion systems are used by many bacterial pathogens for the delivery of virulence factors to the extracellular space or directly into host cells. They are attractive targets for the development of novel anti-virulence drugs as their inactivation would lead to pathogen attenuation or avirulence, followed by clearance of the bacteria by the immune system. This review will present the state of knowledge on the assembly and function of type II, type III and type IV secretion systems in Gram-negative bacteria focusing on insights provided by structural analyses of several key components. The suitability of transcription factors regulating the expression of secretion system components and of ATPases, lytic transglycosylases and protein assembly factors as drug targets will be discussed. Recent progress using innovative in vivo as well as in vitro screening strategies led to a first set of secretion system inhibitors with potential for further development as anti-infectives. The discovery of such inhibitors offers exciting and innovative opportunities to further develop these anti-virulence drugs into monotherapy or in combination with classical antibiotics. Bacterial growth per se would not be inhibited by such drugs so that the selection for mutations causing resistance could be reduced. Secretion system inhibitors may therefore avoid many of the problems associated with classical antibiotics and may constitute a valuable addition to our arsenal for the treatment of bacterial infections.

Phosphatidic Acid Sequesters Sec18p from cis-SNARE Complexes to Inhibit Priming.

PubMed

Starr, Matthew L; Hurst, Logan R; Fratti, Rutilio A

2016-10-01

Yeast vacuole fusion requires the activation of cis-SNARE complexes through priming carried out by Sec18p/N-ethylmaleimide sensitive factor and Sec17p/α-SNAP. The association of Sec18p with vacuolar cis-SNAREs is regulated in part by phosphatidic acid (PA) phosphatase production of diacylglycerol (DAG). Inhibition of PA phosphatase activity blocks the transfer of membrane-associated Sec18p to SNAREs. Thus, we hypothesized that Sec18p associates with PA-rich membrane microdomains before transferring to cis-SNARE complexes upon PA phosphatase activity. Here, we examined the direct binding of Sec18p to liposomes containing PA or DAG. We found that Sec18p preferentially bound to liposomes containing PA compared with those containing DAG by approximately fivefold. Additionally, using a specific PA-binding domain blocked Sec18p binding to PA-liposomes and displaced endogenous Sec18p from isolated vacuoles. Moreover, the direct addition of excess PA blocked the priming activity of isolated vacuoles in a manner similar to chemically inhibiting PA phosphatase activity. These data suggest that the conversion of PA to DAG facilitates the recruitment of Sec18p to cis-SNAREs. Purified vacuoles from yeast lacking the PA phosphatase Pah1p showed reduced Sec18p association with cis-SNAREs and complementation with plasmid-encoded PAH1 or recombinant Pah1p restored the interaction. Taken together, this demonstrates that regulating PA concentrations by Pah1p activity controls SNARE priming by Sec18p. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Characterization of a gene product (Sec53p) required for protein assembly in the yeast endoplasmic reticulum

PubMed Central

1985-01-01

SEC53, a gene that is required for completion of assembly of proteins in the endoplasmic reticulum in yeast, has been cloned, sequenced, and the product localized by cell fractionation. Complementation of a sec53 mutation is achieved with unique plasmids from genomic or cDNA expression banks. These inserts contain the authentic gene, a cloned copy of which integrates at the sec53 locus. An open reading frame in the insert predicts a 29-kD protein with no significant hydrophobic character. This prediction is confirmed by detection of a 28-kD protein overproduced in cells that carry SEC53 on a multicopy plasmid. To follow Sec53p more directly, a LacZ-SEC53 gene fusion has been constructed which allows the isolation of a hybrid protein for use in production of antibody. With such an antibody, quantitative immune decoration has shown that the sec53-6 mutation decreases the level of Sec53p at 37 degrees C, while levels comparable to wild-type are seen at 24 degrees C. An eightfold overproduction of Sec53p accompanies transformation of cells with a multicopy plasmid containing SEC53. Cell fractionation, performed with conditions that preserve the lumenal content of the endoplasmic reticulum (ER), shows Sec53p highly enriched in the cytosol fraction. We suggest that Sec53p acts indirectly to facilitate assembly in the ER, possibly by interacting with a stable ER component, or by providing a small molecule, other than an oligosaccharide precursor, necessary for the assembly event. PMID:3905826

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Code of Federal Regulations, 2013 CFR

2013-01-01

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Code of Federal Regulations, 2014 CFR

2014-01-01

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10 CFR Appendix A to Subpart A of... - Selected Provisions of the Atomic Energy Act of 1954, as Amended, Sec. 141 (42 U.S.C. 2161), Sec...

Code of Federal Regulations, 2012 CFR

2012-01-01

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Code of Federal Regulations, 2011 CFR

2011-01-01

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Code of Federal Regulations, 2010 CFR

2010-01-01

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Landing Characteristics of the Apollo Spacecraft with Deployed Heat Shield Impact Attenuation Systems

NASA Technical Reports Server (NTRS)

Stubbs, Sandy M.

1965-01-01

An experimental investigation was made to determine the landing characteristics of a 1/4-scale dynamic model of the Apollo spacecraft command module using two different active (heat shield deployed prior to landing) landing systems for impact attenuation. One landing system (configuration 1) consisted of six hydraulic struts and eight crushable honeycomb struts. The other landing system (configuration 2), consisted of four hydraulic struts and six strain straps. Tests made on water and the hard clay-gravel composite landing surfaces simulated parachute letdown (vertical) velocities of 23 ft/sec (7.0 m/s) (full scale). Landings made on the sand landing surface simulated vertical velocities of 30 ft/sec (9.1 m/s). Horizontal velocities of from 0 to 50 ft/sec (15 m/s) were simulated. Landing attitudes ranged from -30'degrees to 20 degrees, and the roll attitudes were O degrees, 90 degrees, and 180 degrees. For configuration 1, maximum normal accelerations at the vehicle center of gravity for landings on water, sand, and the hard clay-gravel composite surface were 9g, 20g, and 18g, respectively. The maximum normal center-of-gravity acceleration for configuration 2 which was landed only on the hard clay-gravel landing surface was approximately 19g. Accelerations for configuration 2 were generally equal to or lower than accelerations for configuration 1 and normal.

Warfare between Host Immunity and Bacterial Weapons.

PubMed

Yu, Manda; Lai, Erh-Min

2017-01-11

Bacterial pathogens deploy protein secretion systems to facilitate infection and colonization of their hosts. In this issue of Cell Host & Microbe, Chen et al. (2017) report a new role for a type VI secretion effector in promoting bacterial colonization by preventing inflammasome activation induced by a type III secretion system. Copyright © 2017 Elsevier Inc. All rights reserved.

Trade Secret Law and Information Systems: Can Your Students Keep a Secret?

ERIC Educational Resources Information Center

Willey, Lorrie; Ford, Janet C.; White, Barbara Jo; Clapper, Danial L.

2011-01-01

The impact of intellectual property (IP) law on information systems (IS) professionals in business cannot be overstated. The IS 2010 model curriculum guidelines for undergraduate IS programs stress the importance of information security and knowledge about IP. While copyright and patents are the most well-known types of IP, another, trade secrets,…

Testing limits to airflow perturbation device (APD) measurements.

PubMed

Lopresti, Erika R; Johnson, Arthur T; Koh, Frank C; Scott, William H; Jamshidi, Shaya; Silverman, Nischom K

2008-10-31

The Airflow Perturbation Device (APD) is a lightweight, portable device that can be used to measure total respiratory resistance as well as inhalation and exhalation resistances. There is a need to determine limits to the accuracy of APD measurements for different conditions likely to occur: leaks around the mouthpiece, use of an oronasal mask, and the addition of resistance in the respiratory system. Also, there is a need for resistance measurements in patients who are ventilated. Ten subjects between the ages of 18 and 35 were tested for each station in the experiment. The first station involved testing the effects of leaks of known sizes on APD measurements. The second station tested the use of an oronasal mask used in conjunction with the APD during nose and mouth breathing. The third station tested the effects of two different resistances added in series with the APD mouthpiece. The fourth station tested the usage of a flexible ventilator tube in conjunction with the APD. All leaks reduced APD resistance measurement values. Leaks represented by two 3.2 mm diameter tubes reduced measured resistance by about 10% (4.2 cmH2O.sec/L for control and 3.9 cm H2O.sec/L for the leak). This was not statistically significant. Larger leaks given by 4.8 and 6.4 mm tubes reduced measurements significantly (3.4 and 3.0 cm cmH2O.sec/L, respectively). Mouth resistance measured with a cardboard mouthpiece gave an APD measurement of 4.2 cm H2O.sec/L and mouth resistance measured with an oronasal mask was 4.5 cm H2O.sec/L; the two were not significantly different. Nose resistance measured with the oronasal mask was 7.6 cm H2O.sec/L. Adding airflow resistances of 1.12 and 2.10 cm H2O.sec/L to the breathing circuit between the mouth and APD yielded respiratory resistance values higher than the control by 0.7 and 2.0 cm H2O.sec/L. Although breathing through a 52 cm length of flexible ventilator tubing reduced the APD measurement from 4.0 cm H2O.sec/L for the control to 3.6 cm H2O.sec/L for the tube, the difference was not statistically significant. The APD can be adapted for use in ventilated, unconscious, and uncooperative patients with use of a ventilator tube and an oronasal mask without significantly affecting measurements. Adding a resistance in series with the APD mouthpiece has an additive effect on resistance measurements, and can be used for qualitative calibration. A leak size of at least the equivalent of two 3.2 mm diameter tubes can be tolerated without significantly affecting APD measurements.

Two-Partner Secretion: Combining Efficiency and Simplicity in the Secretion of Large Proteins for Bacteria-Host and Bacteria-Bacteria Interactions

PubMed Central

Guérin, Jeremy; Bigot, Sarah; Schneider, Robert; Buchanan, Susan K.; Jacob-Dubuisson, Françoise

2017-01-01

Initially identified in pathogenic Gram-negative bacteria, the two-partner secretion (TPS) pathway, also known as Type Vb secretion, mediates the translocation across the outer membrane of large effector proteins involved in interactions between these pathogens and their hosts. More recently, distinct TPS systems have been shown to secrete toxic effector domains that participate in inter-bacterial competition or cooperation. The effects of these systems are based on kin vs. non-kin molecular recognition mediated by specific immunity proteins. With these new toxin-antitoxin systems, the range of TPS effector functions has thus been extended from cytolysis, adhesion, and iron acquisition, to genome maintenance, inter-bacterial killing and inter-bacterial signaling. Basically, a TPS system is made up of two proteins, the secreted TpsA effector protein and its TpsB partner transporter, with possible additional factors such as immunity proteins for protection against cognate toxic effectors. Structural studies have indicated that TpsA proteins mainly form elongated β helices that may be followed by specific functional domains. TpsB proteins belong to the Omp85 superfamily. Open questions remain on the mechanism of protein secretion in the absence of ATP or an electrochemical gradient across the outer membrane. The remarkable dynamics of the TpsB transporters and the progressive folding of their TpsA partners at the bacterial surface in the course of translocation are thought to be key elements driving the secretion process. PMID:28536673

Comparison of Three Wind Measuring Systems for Flight Test

NASA Technical Reports Server (NTRS)

Teets, Edward H., Jr.; Harvey, Philip O.

2000-01-01

A preliminary field test of the accuracy of wind velocity measurements obtained using global positioning system-tracked rawinsonde balloons has been performed. Wind comparisons have been conducted using global positioning system (GPS) and radio automatic theodolite sounder (RATS) rawinsondes and a high-precision range instrumentation radar-tracked reflector. Wind velocity differences between the GPS rawinsondes and the radar were significantly less than between the RATS rawinsondes and the radar. These limited test results indicate a root-mean-square wind velocity difference from 4.98 kn (2.56 m/sec) for the radar and RATS to 1.09 kn (0.56 m/sec) for the radar and GPS. Differences are influenced by user reporting requirements, data processing techniques, and the inherent tracking accuracies of the system. This brief field test indicates that the GPS sounding system tracking data are more precise than the RATS system. When high-resolution wind data are needed, use of GPS rawinsonde systems can reduce the burden on range radar operations.

The Spanish Society of Soil Science: Main projects and activities developed during the last years

NASA Astrophysics Data System (ADS)

Porta, Jaume; Mataix-Solera, Jorge; Ortiz-Bernad, Irene; Arbelo, Carmen D.; Díaz-Raviña, Montserrat; Badía, David; Alcañiz, Josep M.; Santos, Fernando; Hermosin, M. Carmen; Barral, M. Teresa

2017-04-01

The Spanish Society of Soil Science (in Spanish: Sociedad Española de la Ciencia del Suelo, SECS) was founded in 1947 by the Spanish National Research Council (CSIC) to promote cohesion and collaboration between soil science professionals, and with an innovative spirit and a willingness to serve the society. The objectives are: to promote the study, knowledge, research and protection of the soil; to spread, from a scientific point of view, the role played by the soil in favour of society, through ecosystem services such as the production of food and raw materials; The SECS also focus on the protection of other habitats and the conservation of our archaeological heritage; and to preserve knowledge of the soil, its management and use, both from the production and environmental point of view, leading to the optimization of its capabilities. The activities and services of the SECS are accessible on the web site www.secs.com.es, which is continually updated. In this contribution, we will show some examples of recent projects and activities developed by the SECS such as: the edition of the Spanish Journal of Soil Science (SJSS) since 2011; books like the white book on "Tratamiento del suelo en los libros de texto de ESO y Bachillerato en España" available in http://www.secs.com.es/archivo/libro-suelo.pdf, in which the term "Soil" is analysed in many secondary school books (152) in the Spanish education system; conferences, courses, exhibitions, expositions, calendars, the comic "Vivir en el suelo" in diferent languages, and diverse material to promote and disseminate the importance of the soil to the society; and last but not least, the Multingual Soil Science Dictionary (Spanish, Catalan, Gallician and Potuguese with translations in English and French (in process)) available online: http://cit.iec.cat/GLOSECS/inici.html. The promotion of student teams for soil science contests, the SECS Award to attend the Simposio Latinoamericano de Enseñanza y Educación en Ciencia del Suelo (SLCS, Quito, 2016) and other activities, such as incentives for young students, are also some of the strategies to promote the soil, together with an annual award to the best PhD thesis supervised by SECS members. These and other examples will be shown during the session with the main aim of exchanging ideas among soil societies to promote the soil. Access in www.secs.com.es. Acknowledgements: to the members of the Spanish Society of Soil Science.

Solution Structure of Homology Region (HR) Domain of Type II Secretion System*

PubMed Central

Gu, Shuang; Kelly, Geoff; Wang, Xiaohui; Frenkiel, Tom; Shevchik, Vladimir E.; Pickersgill, Richard W.

2012-01-01

The type II secretion system of Gram-negative bacteria is important for bacterial pathogenesis and survival; it is composed of 12 mostly multimeric core proteins, which build a sophisticated secretion machine spanning both bacterial membranes. OutC is the core component of the inner membrane subcomplex thought to be involved in both recognition of substrate and interaction with the outer membrane secretin OutD. Here, we report the solution structure of the HR domain of OutC and explore its interaction with the secretin. The HR domain adopts a β-sandwich-like fold consisting of two β-sheets each composed of three anti-parallel β-strands. This structure is strikingly similar to the periplasmic region of PilP, an inner membrane lipoprotein from the type IV pilus system highlighting the common evolutionary origin of these two systems and showing that all the core components of the type II secretion system have a structural or sequence ortholog within the type IV pili system. The HR domain is shown to interact with the N0 domain of the secretin. The importance of this interaction is explored in the context of the functional secretion system. PMID:22253442

Influence of high-resolution surface databases on the modeling of local atmospheric circulation systems

NASA Astrophysics Data System (ADS)

Paiva, L. M. S.; Bodstein, G. C. R.; Pimentel, L. C. G.

2013-12-01

Large-eddy simulations are performed using the Advanced Regional Prediction System (ARPS) code at horizontal grid resolutions as fine as 300 m to assess the influence of detailed and updated surface databases on the modeling of local atmospheric circulation systems of urban areas with complex terrain. Applications to air pollution and wind energy are sought. These databases are comprised of 3 arc-sec topographic data from the Shuttle Radar Topography Mission, 10 arc-sec vegetation type data from the European Space Agency (ESA) GlobCover Project, and 30 arc-sec Leaf Area Index and Fraction of Absorbed Photosynthetically Active Radiation data from the ESA GlobCarbon Project. Simulations are carried out for the Metropolitan Area of Rio de Janeiro using six one-way nested-grid domains that allow the choice of distinct parametric models and vertical resolutions associated to each grid. ARPS is initialized using the Global Forecasting System with 0.5°-resolution data from the National Center of Environmental Prediction, which is also used every 3 h as lateral boundary condition. Topographic shading is turned on and two soil layers with depths of 0.01 and 1.0 m are used to compute the soil temperature and moisture budgets in all runs. Results for two simulated runs covering the period from 6 to 7 September 2007 are compared to surface and upper-air observational data to explore the dependence of the simulations on initial and boundary conditions, topographic and land-use databases and grid resolution. Our comparisons show overall good agreement between simulated and observed data and also indicate that the low resolution of the 30 arc-sec soil database from United States Geological Survey, the soil moisture and skin temperature initial conditions assimilated from the GFS analyses and the synoptic forcing on the lateral boundaries of the finer grids may affect an adequate spatial description of the meteorological variables.

Characterization of experimentally induced, nonaflatoxigenic variant strains of Aspergillus parasiticus.

PubMed Central

Kale, S P; Cary, J W; Bhatnagar, D; Bennett, J W

1996-01-01

Six previously isolated, nonaflatoxigenic variants of Aspergillus parasiticus, designated sec mutants, were characterized morphologically by electron microscopy, biochemically by biotransformation studies with an aflatoxin precursor, and genetically by Northern (RNA) hybridization analysis of aflatoxin biosynthetic gene transcripts. Scanning electron micrographs clearly demonstrated that compared with the parental sec+ forms, the variant sec forms had an abundance of vegetative mycelia, orders of magnitude reduced number of conidiophores and conidia, and abnormal metulae. Conidiospores were detected in sec cultures only at higher magnifications (x 500), in contrast to the sec+ (wild-type) strain, in which abundant conidiospores (masking the vegetative mycelia) were observed at even lower magnifications (x 300). All sec+ forms, but none of the sec forms, showed bioconversion of sterigmatocystin to aflatoxins. Northern blots probed with pathway genes demonstrated lack of expression of both the aflatoxin biosynthetic pathway structural (nor-1 and omtA) and regulatory (aflR) genes in the sec forms; PCR and Southern hybridization analysis confirmed the presence of the genes in the sec genomes. Thus, the loss of aflatoxigenic capabilities in the sec form is correlated with alterations in the conidial morphology of the fungus, suggesting that the regulation of aflatoxin synthesis and conidiogenesis may be interlinked. PMID:8795232

Sec34 is implicated in traffic from the endoplasmic reticulum to the Golgi and exists in a complex with GTC-90 and ldlBp.

PubMed

Loh, Eva; Hong, Wanjin

2002-06-14

Sec34p/Grd20p has been implicated in endoplasmic reticulum (ER)-to-Golgi transport and/or post-Golgi trafficking events and exists in a protein complex consisting of at least eight subunits in yeast. Although the mammalian counterpart (Sec34) of Sec34p has been molecularly identified, its role and interacting partners remain undefined. In this study, we have prepared antibodies specifically against the recombinant N-terminal fragment of Sec34 that recognize a polypeptide of about 93 kDa and label the Golgi apparatus. In a well-characterized semi-intact cell assay that reconstitutes transport of the envelope glycoprotein (VSVG) of vesicular stomatitis virus from the ER to the Golgi apparatus, anti-Sec34 antibodies inhibited the transport in a dose-dependent manner. The inhibition by anti-Sec34 antibodies could be neutralized by a noninhibitory amount of the antigen. Large-scale immunoprecipitation of rat liver cytosol with immobilized anti-Sec34 antibodies has co-immunoprecipitated GTC-90 and ldlBp, two peripheral Golgi proteins previously shown to exist in separate protein complexes. Two mammalian homologues (Dor1 and Cod1) of the yeast Sec34 complex were similarly recovered in the Sec34 immunoprecipitates. When expressed in transfected cells, epitope-tagged ldlCp and Cod2 were co-immunoprecipitated with anti-Sec34 antibodies with efficiencies comparable to that observed for tagged ldlBp, Dor1, and Cod1. Direct interactions of Sec34 with ldlBp and ldlCp were further demonstrated in vitro. These results suggest that Sec34, GTC-90, and ldlBp/ldlCp are part of the same protein complex(es) that regulates diverse aspects of Golgi function, including transport from the ER to the Golgi apparatus.

Selenocysteine in proteins-properties and biotechnological use.

PubMed

Johansson, Linda; Gafvelin, Guro; Arnér, Elias S J

2005-10-30

Selenocysteine (Sec), the 21st amino acid, exists naturally in all kingdoms of life as the defining entity of selenoproteins. Sec is a cysteine (Cys) residue analogue with a selenium-containing selenol group in place of the sulfur-containing thiol group in Cys. The selenium atom gives Sec quite different properties from Cys. The most obvious difference is the lower pK(a) of Sec, and Sec is also a stronger nucleophile than Cys. Proteins naturally containing Sec are often enzymes, employing the reactivity of the Sec residue during the catalytic cycle and therefore Sec is normally essential for their catalytic efficiencies. Other unique features of Sec, not shared by any of the other 20 common amino acids, derive from the atomic weight and chemical properties of selenium and the particular occurrence and properties of its stable and radioactive isotopes. Sec is, moreover, incorporated into proteins by an expansion of the genetic code as the translation of selenoproteins involves the decoding of a UGA codon, otherwise being a termination codon. In this review, we will describe the different unique properties of Sec and we will discuss the prerequisites for selenoprotein production as well as the possible use of Sec introduction into proteins for biotechnological applications. These include residue-specific radiolabeling with gamma or positron emitters, the use of Sec as a reactive handle for electophilic probes introducing fluorescence or other peptide conjugates, as the basis for affinity purification of recombinant proteins, the trapping of folding intermediates, improved phasing in X-ray crystallography, introduction of 77Se for NMR spectroscopy, or, finally, the analysis or tailoring of enzymatic reactions involving thiol or oxidoreductase (redox) selenolate chemistry.

A western boundary current east of New Caledonia: Observed characteristics

NASA Astrophysics Data System (ADS)

Gasparin, Florent; Ganachaud, Alexandre; Maes, Christophe

2011-09-01

Waters from the South Equatorial Current (SEC), the northern branch of the South Pacific subtropical gyre, are a major supply of heat to the equatorial warm pool, and have an important contribution to climate variability and ENSO which motivated the Southwest Pacific Ocean and Climate Experiment (SPICE, CLIVAR/WCRP). Initially a broad westward current extending from the equator to 30°S, the SEC splits upon arriving at the major islands and archipelagoes of Fiji (18°S, 180°E), Vanuatu (16°S, 168°E), and New Caledonia (22°S, 165°E), resulting in a complex system of western boundary currents and zonal jets that feed the Coral and Solomon Seas. We focus here on the formation of one specific jet feeding the Coral Sea, the North Caledonian Jet (NCJ). Using a combination of recent oceanographic cruises, we describe the ocean circulation to the northeast of New Caledonia, where the SEC forms a western boundary current that ultimately becomes the NCJ. This current, which we document for the first time and propose to refer to as the East Caledonian Current (ECC), has its core located 10-100 km off the east coast of New Caledonia, and extends vertically to at least 1000 m depth. Water mass properties show continuous westward transports through the ECC, from the SEC to the NCJ in both the South Pacific Tropical Waters in the thermocline and Antarctic Intermediate Waters near 700 m depth. The ECC extends about 100 km horizontally; its average 0-1000 m transport was estimated at 14.5±3 Sv off the north tip of the New Caledonian reef, with a maximum of 20 Sv in May 2010. South of that the upstream branch of the ECC east of the Loyalty is close to 8 Sv suggesting an important additional contribution from central Pacific waters carried by the SEC at 16°S and diverted to our region through the western boundary current system east of Vanuatu.

Microcomputer control of infrared detector arrays used in direct imaging and in Fabry-Perot spectroscopy

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

Rossano, G.S.

1989-02-01

A microcomputer based data acquisition system has been developed for astronomical observing with two-dimensional infrared detector arrays operating at high pixel rates. The system is based on a 16-bit 8086/8087 microcomputer operating at 10 MHz. Data rates of up to 560,000 pixels/sec from arrays of up to 4096 elements are supported using the microcomputer system alone. A hardware co-adder the authors are developing permits data accumulation at rates of up to 1.67 million pixels/sec in both staring and chopped data acquisition modes. The system has been used for direct imaging and for data acquisition in a Fabry-Perot Spectrometer developed bymore » NRL. The hardware is operated using interactive software which supports the several available modes of data acquisition, and permits data display and reduction during observing sessions.« less

Mechanism of Occurring Over-Voltage Phenomena in Distributed Power System on Energization of Transformers

NASA Astrophysics Data System (ADS)

Nakachi, Yoshiki; Ueda, Fukashi; Kajikawa, Takuya; Amau, Tooru; Kameyama, Hirokazu; Ito, Hisanori

This paper verifies the mechanism of occurring over voltage phenomena in the distributed power system on energizing the transformer. This over-voltage, which is observed at the actual distributed power system, with heavy inrush current is found to occur at about 0.1-0.2sec after the energizing and continue for a duration of more than 0.1[sec]. There is a concern that this over-voltage may operate the protection relay and deteriorate the insulation of apparatus. It is basically caused by the resonance between the shunt capacitors and saturated/unsaturated magnetizing inductance of transformer, system inductance. By using analytical formulation of a simple equivalent circuit, its mechanism has been verified through simulations carried out by using EMTP. Moreover, the sympathetic interaction between transformers is prolonged the duration of the over-voltage by the field test data is discussed in this paper.

A CDMA synchronisation scheme

NASA Technical Reports Server (NTRS)

Soprano, C.

1993-01-01

CDMA (Code Division Multiple Access) is known to decrease inter-service interference in Satellite Communication Systems. Its performance is increased by chip quasi-synchronous operation which virtually eliminates the self-noise; however, the theory shows that the time error on the synchronization has to be kept at less than one tenth of a chip which, for 1 Mchip/sec. spreading rate, corresponds to 10(exp -7) sec. This, on the return-link, may only be achieved by means of a closed loop control system which, for mobile communication systems, has to be capable of autonomous operation. Until now some results have been reported on the feasibility of chip quasi-synchronous operation for mobile communication systems only including satellites on GEO (Geostationary Earth Orbit). In what follows, the basic principles are exposed, and results are presented showing how low chip synchronism error may be achieved by means of an autonomous control loop operating through satellites on any Earth orbit.

Controlled Bidirectional Quantum Secure Direct Communication

PubMed Central

Chou, Yao-Hsin; Lin, Yu-Ting; Zeng, Guo-Jyun; Lin, Fang-Jhu; Chen, Chi-Yuan

2014-01-01

We propose a novel protocol for controlled bidirectional quantum secure communication based on a nonlocal swap gate scheme. Our proposed protocol would be applied to a system in which a controller (supervisor/Charlie) controls the bidirectional communication with quantum information or secret messages between legitimate users (Alice and Bob). In this system, the legitimate users must obtain permission from the controller in order to exchange their respective quantum information or secret messages simultaneously; the controller is unable to obtain any quantum information or secret messages from the decoding process. Moreover, the presence of the controller also avoids the problem of one legitimate user receiving the quantum information or secret message before the other, and then refusing to help the other user decode the quantum information or secret message. Our proposed protocol is aimed at protecting against external and participant attacks on such a system, and the cost of transmitting quantum bits using our protocol is less than that achieved in other studies. Based on the nonlocal swap gate scheme, the legitimate users exchange their quantum information or secret messages without transmission in a public channel, thus protecting against eavesdroppers stealing the secret messages. PMID:25006596

Improvements To Micro Contact Performance And Reliability

DTIC Science & Technology

2016-12-22

BAKE : □ 1 min 110°C hot plate bake OPTIONAL - For Grayscale Lithography: 1818 COAT: □ Flood wafer with 1818 □ 4 sec spread at 500 rpm...30 sec spin at 4,000 rpm, ramp=200 □ 75 sec 110°C hot plate bake GRAYSCALE PATTERNING: □ Follow grayscale patterning process for patterns...8217 □ 2 min 200°C hot plate bake 1818 COAT: □ Flood wafer with 1818 □ 4 sec spread at 500 rpm □ 30 sec spin at 4,000 rpm, ramp=200 □ 75 sec 110

A 1055 ft/sec impact test of a two foot diameter model nuclear reactor containment system without fracture

NASA Technical Reports Server (NTRS)

Puthoff, R. L.

1972-01-01

A study to determine the feasibility of containing the fission products of a mobile reactor in the event of an impact is presented. The model simulated the reactor core, energy absorbing gamma shielding, neutron shielding and the containment vessel. It was impacted against an 18,000 pound reinforced concrete block at 1055 ft/sec. The model was significantly deformed and the concrete block demolished. No leaks were detected nor were any cracks observed in the model after impact.

High performance quarter-inch cartridge tape systems

NASA Technical Reports Server (NTRS)

Schwarz, Ted

1993-01-01

Within the established low cost structure of Data Cartridge drive technology, it is possible to achieve nearly 1 terrabyte (10(exp 12)) of data capacity and more than 1 Gbit/sec (greater than 100 Mbytes/sec) transfer rates. The desirability to place this capability within a single cartridge will be determined by the market. The 3.5 in. or smaller form factor may suffice to serve both the current Data Cartridge market and a high performance segment. In any case, Data Cartridge Technology provides a strong sustainable technology growth path in the 21st century.

Centripetal myosin redistribution in thrombin-stimulated platelets. Relationship to platelet Factor 4 secretion.

PubMed

Painter, R G; Ginsberg, M H

1984-11-01

We have examined the F-actin and myosin distribution in resting and thrombin-activated platelets by double label immunofluorescence microscopy. In resting, discoid platelets, F-actin and myosin staining was distributed in a diffuse pattern throughout the interior of the cell with slight accentuation at the cell periphery. In contrast, platelet factor 4 antigen (PF4) was more centrally localized in a fine punctate distribution which is consistent with its localization in alpha-granules. Within 5 sec after thrombin stimulation both F-actin and myosin staining were increased at the periphery of the now spherical platelets. Subsequently, a myosin-containing spherical structure decreased in diameter closely surrounding a phase-dense central zone. In contrast, F-actin staining continued to be accentuated at the cell periphery and was prominent in filopodia and blebs. As previously shown, PF4 staining was localized after 30 sec within large intracellular masses that corresponded to closed vacuolar structures at the ultrastructural level. Morphometric analysis of electron micrographs showed that formation of these vacuolar structures kinetically paralleled alpha-granule disappearance and preceded PF4 release. These PF4-containing structures translocated to the cell periphery after 1-3 min, where they appeared to fuse with the plasma membrane. Ultrastructural analysis of thin sections showed that the myosin-rich spherical structure spatially and temporally correlated with a band of microfilaments that closely surrounded the organelle-rich central zone of the cell. Morphometric analysis of these micrographs showed that the absolute volume of this central zone decreased with time after thrombin addition, showing a significant change after 15 sec and reaching a maximum value after 3-5 min. Changes in the volume of this compartment kinetically preceded PF4 release. On the basis of these data, we propose that an actomyosin contractile force is generated which centripetally redistributes the myosinrich structure and organelle zone. Conceivably this inward force may not only accelerate granule-granule fusion to form intracellular secretory vacuoles, but may also provide aid in their extrusion toward the platelet plasma membrane.

A Study of relationship between frailty and physical performance in elderly women

PubMed Central

Jeoung, Bog Ja; Lee, Yang Chool

2015-01-01

Frailty is a disorder of multiple inter-related physiological systems. It is unclear whether the level of physical performance factors can serve as markers of frailty and a sign. The purpose of this study was to examine the relationship between frailty and physical performance in elderly women. One hundred fourteen elderly women participated in this study, their aged was from 65 to 80. We were measured 6-min walk test, grip-strength, 30-sec arm curl test, 30-sec chair stand test, 8 foot Up- and Go, Back scratch, chair sit and reach, unipedal stance, BMI, and the frailty with questionnaire. The collected data were analyzed by descriptive statistics, frequencies, correlation analysis, ANOVA, and simple liner regression using the IBM 21. SPSS program. In results, statistic tests showed that there were significant differences between frailty and 6-min walk test, 30-sec arm curl test, 30-sec chair stand test, grip-strength, Back scratch, and BMI. However, we did not find significant differences between frailty and 8 foot Up- and Go, unipedal stance. When the subjects were divided into five groups according to physical performance level, subjects with high 6-min walk, 30-sec arm curl test, chair sit and reach test, and high grip strength had low score frailty. Physical performance factors were strongly associated with decreased frailty, suggesting that physical performance improvements play an important role in preventing or reducing the frailty. PMID:26331137

Geographic Information System and Remote Sensing Approach with Hydrologic Rational Model for Flood Event Analysis in Jakarta

NASA Astrophysics Data System (ADS)

Aditya, M. R.; Hernina, R.; Rokhmatuloh

2017-12-01

Rapid development in Jakarta which generates more impervious surface has reduced the amount of rainfall infiltration into soil layer and increases run-off. In some events, continuous high rainfall intensity could create sudden flood in Jakarta City. This article used rainfall data of Jakarta during 10 February 2015 to compute rainfall intensity and then interpolate it with ordinary kriging technique. Spatial distribution of rainfall intensity then overlaid with run-off coefficient based on certain land use type of the study area. Peak run-off within each cell resulted from hydrologic rational model then summed for the whole study area to generate total peak run-off. For this study area, land use types consisted of 51.9 % industrial, 37.57% parks, and 10.54% residential with estimated total peak run-off 6.04 m3/sec, 0.39 m3/sec, and 0.31 m3/sec, respectively.

Rotor redesign for a highly loaded 1800 ft/sec tip speed fan, 2

NASA Technical Reports Server (NTRS)

Bolt, C. R.

1980-01-01

Tests were conducted on a 0.5 hub/tip ratio single-stage fan designed to produce a pressure ratio of 2.280 at an efficiency of 83.8 percent with a rotor tip speed of 548.6 m/sec (1800 ft/sec). The rotor was designed utilizing a quasi three dimensional design system and four-part, multiple-circular-arc airfoil sections. The rotor is the third in a series of single-stage fans that have included a precompression airfoil design and a multiple-circular-arc airfoil design. The stage achieved a peak efficiency of 82.8 percent after performance had deteriorated by 0.6 of a point. The design mass flow was achieved at the peak efficiency point, and the stage total pressure ratio was 2.20, which is lower than the design goal of 2.28. The surge margin of 13% from the peak efficiency point exceeded the design goal of 7%.

NASA electrothermal auxiliary propulsion technology

NASA Technical Reports Server (NTRS)

Stone, J. R.

1986-01-01

Electrothermal auxiliary propulsion systems provide high performance options which can have major mission benefits. There are several electrothermal concepts which offer a range of characteristics and benefits. Resistojets are the highest thrust to power option and are currently operational at mission average values of specific impulse, I sub sp approximately 295 sec. Long life, multipropellant resistojets are being developed for the space station, and resistojet technology advancements are being pursued to improve the I sub sp by more than 20 percent for resistojets used in satellite applications. Direct current arcjets have the potential of I sub sp over 400 sec with storable propellants and should provide over 1000 sec with hydrogen. Advanced concepts are being investigated to provide high power density options and possible growth to primary propulsion applications. Broad based experimental and analytical research and technology programs of NASA are summarized and recent significant advances are reviewed.

29 CFR 5.1 - Purpose and scope.

Code of Federal Regulations, 2010 CFR

2010-07-01

..., Health Statistics, and Medical Libraries Act of 1974 (sec. 107, see sec. 308(h)(2) thereof, 88 Stat. 370... 1965 (sec. 712; 79 Stat. 575 as amended; 42 U.S.C. 3222). 42. Juvenile Delinquency Prevention Act (sec...

Agonal sequences in a filmed suicidal hanging: analysis of respiratory and movement responses to asphyxia by hanging.

PubMed

Sauvageau, Anny; Racette, Stéphanie

2007-07-01

The forensic literature on the pathophysiology of human hanging is still limited. Therefore, forensic pathologists often feel uncomfortable when confronted with related questions. Here presented is the filmed suicidal hanging of a 37-year-old man. This recording allows a unique analysis of agonal movement sequences: loss of consciousness (13 sec), convulsions (15 sec), decortication rigidity (21 sec), decerebration rigidity (46 sec), second decortication rigidity (1 min 11 sec), loss of muscle tone, (1 min 38 sec) and last isolated muscle movement (4 min 10 sec). As for respiratory responses, very deep respiratory attempts started at 20 sec. Respiratory movements progressively decreased and completely stopped at 2 min. Despite the fact that extending the presented data on all cases of hanging asphyxia would be a mistake, this case gives a very interesting insight into movement and respiratory response to asphyxia by hanging.

Secret Sharing and Shared Information

NASA Astrophysics Data System (ADS)

Rauh, Johannes

2017-11-01

Secret sharing is a cryptographic discipline in which the goal is to distribute information about a secret over a set of participants in such a way that only specific authorized combinations of participants together can reconstruct the secret. Thus, secret sharing schemes are systems of variables in which it is very clearly specified which subsets have information about the secret. As such, they provide perfect model systems for information decompositions. However, following this intuition too far leads to an information decomposition with negative partial information terms, which are difficult to interpret. One possible explanation is that the partial information lattice proposed by Williams and Beer is incomplete and has to be extended to incorporate terms corresponding to higher order redundancy. These results put bounds on information decompositions that follow the partial information framework, and they hint at where the partial information lattice needs to be improved.

Secret Key Generation via a Modified Quantum Secret Sharing Protocol

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

Smith IV, Amos M; Evans, Philip G; Lawrie, Benjamin J

We present and experimentally show a novel protocol for distributing secret information between two and only two parties in a N-party single-qubit Quantum Secret Sharing (QSS) system. We demonstrate this new algorithm with N = 3 active parties over 6km of telecom. ber. Our experimental device is based on the Clavis2 Quantum Key Distribution (QKD) system built by ID Quantique but is generalizable to any implementation. We show that any two out of the N parties can build secret keys based on partial information from each other and with collaboration from the remaining N > 2 parties. This algorithm allowsmore » for the creation of two-party secret keys were standard QSS does not and signicantly reduces the number of resources needed to implement QKD on a highly connected network such as the electrical grid.« less

Double mutation of cell wall proteins CspB and PBP1a increases secretion of the antibody Fab fragment from Corynebacterium glutamicum

PubMed Central

2014-01-01

Background Among other advantages, recombinant antibody-binding fragments (Fabs) hold great clinical and commercial potential, owing to their efficient tissue penetration compared to that of full-length IgGs. Although production of recombinant Fab using microbial expression systems has been reported, yields of active Fab have not been satisfactory. We recently developed the Corynebacterium glutamicum protein expression system (CORYNEX®) and demonstrated improved yield and purity for some applications, although the system has not been applied to Fab production. Results The Fab fragment of human anti-HER2 was successfully secreted by the CORYNEX® system using the conventional C. glutamicum strain YDK010, but the productivity was very low. To improve the secretion efficiency, we investigated the effects of deleting cell wall-related genes. Fab secretion was increased 5.2 times by deletion of pbp1a, encoding one of the penicillin-binding proteins (PBP1a), mediating cell wall peptidoglycan (PG) synthesis. However, this Δpbp1a mutation did not improve Fab secretion in the wild-type ATCC13869 strain. Because YDK010 carries a mutation in the cspB gene encoding a surface (S)-layer protein, we evaluated the effect of ΔcspB mutation on Fab secretion from ATCC13869. The Δpbp1a mutation showed a positive effect on Fab secretion only in combination with the ΔcspB mutation. The ΔcspBΔpbp1a double mutant showed much greater sensitivity to lysozyme than either single mutant or the wild-type strain, suggesting that these mutations reduced cell wall resistance to protein secretion. Conclusion There are at least two crucial permeability barriers to Fab secretion in the cell surface structure of C. glutamicum, the PG layer, and the S-layer. The ΔcspBΔpbp1a double mutant allows efficient Fab production using the CORYNEX® system. PMID:24731213

An Analysis of Alternatives to Verbal FM Radio Tactical Command and Control Communications

DTIC Science & Technology

1975-06-06

specialist’s ,, , nwarted by these means even more reliablv than by conventional weapons.16 renaoiy an enemy attack can be th1 Marshal Vasili ...Mort Pit Squad(4) FDC F0(3) LDR FAC Bn LN0(2) Conmel Cff Survl Sec Sgt Commo Pit Gnd Survl Sec(6) Spt Pit Co Maint Sec AVLB Sec Med Sec Bn

76 FR 2609 - Proposed Amendment of Class E Airspace; Pueblo, CO

Federal Register 2010, 2011, 2012, 2013, 2014

2011-01-14

...]21[sec] N., long. 104[deg]29[min]47[sec] W.) That airspace extending upward from 700 feet above the... Memorial Airport clockwise between the 070[deg] and 133[deg] bearing from the airport; that airspace...., long. 105[deg]00[min]02[sec] W.; to lat. 38[deg]09[min]25[sec] N., long. 105[deg]08[min]06[sec] W.; to...

40 CFR Appendix I to Part 600 - Highway Fuel Economy Driving Schedule (Applicable to 1978 and Later Model Year Automobiles)

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 30 2011-07-01 2011-07-01 false Highway Fuel Economy Driving Schedule (Applicable to 1978 and Later Model Year Automobiles) I Appendix I to Part 600 Protection of Environment... to 1978 and Later Model Year Automobiles) [ speed (mph) vs time (sec) ] SEC MPH SEC MPH SEC MPH SEC...

A Virtual Audio Guidance and Alert System for Commercial Aircraft Operations

NASA Technical Reports Server (NTRS)

Begault, Durand R.; Wenzel, Elizabeth M.; Shrum, Richard; Miller, Joel; Null, Cynthia H. (Technical Monitor)

1996-01-01

Our work in virtual reality systems at NASA Ames Research Center includes the area of aurally-guided visual search, using specially-designed audio cues and spatial audio processing (also known as virtual or "3-D audio") techniques (Begault, 1994). Previous studies at Ames had revealed that use of 3-D audio for Traffic Collision Avoidance System (TCAS) advisories significantly reduced head-down time, compared to a head-down map display (0.5 sec advantage) or no display at all (2.2 sec advantage) (Begault, 1993, 1995; Begault & Pittman, 1994; see Wenzel, 1994, for an audio demo). Since the crew must keep their head up and looking out the window as much as possible when taxiing under low-visibility conditions, and the potential for "blunder" is increased under such conditions, it was sensible to evaluate the audio spatial cueing for a prototype audio ground collision avoidance warning (GCAW) system, and a 3-D audio guidance system. Results were favorable for GCAW, but not for the audio guidance system.

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

Fox, John

A 4.2 GS/sec. beam excitation system with accelerator synchronization and power stages is described. The system is capable of playing unique samples (32 samples/bunch) for 15,000 turns on selected bunch(es) in the SPS in syn- chronism with the injection and acceleration cycle. The purpose of the system is to excite internal modes of single-bunch vertical motion, and study the bunch dynamics in the presence of developing Electron cloud or TMCI effects. The system includes a synchronized master oscillator, SPS timing functions, an FPGA based arbitrary waveform generator, 4.2 GS/sec. D/A system and four 80W 20-1000 MHz amplifiers driving a taperedmore » stripline pickup/kicker. A software GUI allows specification of various modulation signals, selection of bunches and turns to excite, while a remote control interface allows simple control/monitoring of the RF power stages located in the tunnel. The successful use of this system for SPS MD measurements in 2011 is a vital proof-of-principle for wideband feedback using similar functions to correct the beam motion.« less

An Analysis of Perturbed Quantization Steganography in the Spatial Domain

DTIC Science & Technology

2005-03-01

72 4.2.3 Hide v2.1 Steganographic Software & the Desaturate Function. ..................... 73 4.2.4 Studying the Effect of the Secret Message...Effect of the Secret Message Payload.......................................... 84 4.3.4 Performance Comparison of the Three Steganographic Systems...Epsilon .......................................................................... 89 4.4 Secrets for the Secret ............................................................................................... 90

Verifying Secrets and Relative Secrecy

DTIC Science & Technology

2000-01-01

Systems that authenticate a user based on a shared secret (such as a password or PIN) normally allow anyone to query whether the secret is a given...value. For example, an ATM machine allows one to ask whether a string is the secret PIN of a (lost or stolen) ATM card. Yet such queries are prohibited

A Bacterial Pathogen uses Distinct Type III Secretion Systems to Alternate between Host Kingdom

USDA-ARS?s Scientific Manuscript database

Gram-negative bacterial pathogens of eukaryotes often secrete proteins directly into host cells via a needle-like protein channel called a ‘type III secretion system’ (T3SS). Bacteria that are adapted to either animal or plant hosts use phylogenetically distinct T3SSs for secreting proteins. Here, ...

Physics Design Considerations for Diagnostic X Electron Beam Transport System

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

Chen, Y-J

2000-04-10

The Diagnostic X (D-X) beamlines will transport the DARHT-II beam from the end of the accelerator to the Diagnostic X firing point providing four lines of sight for x-ray radiography. The design goal for the Diagnostic X beamline is to deliver four x-ray pulses with the DARHT-II dose format and time integrated spot size on each line of sight. The D-X beamline's final focus should be compatible with a range of first conjugates from 1 m-5 m. Furthermore, the D-X beamline operational parameters and the beamline layout should not preclude a possible upgrade to additional lines of sight. The DARHT-IImore » accelerator is designed to deliver beams at a rate of 1 pulse per minute or less. Tuning the D-X beamline with several hundred optical elements would be time consuming. Therefore, minimizing the required number of tuning shots for the D-X beamline is also an important design goal. Many different beamline configurations may be able to accomplish these design objectives, and high beam quality (i.e., high current and low emittance) must be maintained throughout the chosen beamline configuration in order to achieve the DARHT-II x-ray dose format. In general, the longer the distance a beam travels, the harder it is to preserve the beam quality. Therefore, from the point of view of maintaining beam quality, it is highly desirable to minimize the beamline length. Lastly, modification to the DARHT-II building and the downstream transport should be minimized. Several processes can degrade beam quality by increasing the beam emittance, increasing the time-varying transverse beam motion, creating a beam halo, or creating a time-varying beam envelope. In this report, we consider those processes in the passive magnet lattice beamline and indicate how they constrain the beamline design. The physics design considerations for the active components such as the kicker system will be discussed in Ref. 2. In Sec. I, we discuss how beam emittance affects the x-ray forward dose. We also establish a physics design goal for the emittance growth budget. In Sec. II, we discuss how the conductivity and size of the beam pipe affects the transverse beam motion. We also discuss the emittance growth arise from the beam centroid offset. In Sec. III, we discuss the background gas focusing effects and establish the vacuum requirements. In Sec. IV, we consider the emittance growth in a bend. In Sec. V, we discuss the misalignment and corkscrew motion. The design specifications for misalignment are established. In Secs. VI and VII, we discuss the design objectives on how to extract beams from the DARHT-II beamline and how to minimize the tuning shots. The integrated spot size and final focusing are discussed in Sec. VIII. A conclusion will be presented in Sec. IX.« less

Repeated sprint ability related to recovery time in young soccer players.

PubMed

Padulo, J; Tabben, M; Ardigò, L P; Ionel, M; Popa, C; Gevat, C; Zagatto, A M; Dello Iacono, A

2015-01-01

This study aimed to describe the influence of recovery duration during a repeated sprint ability (RSA) test (6 × 40 m) by investigating a number of variables, such as general performance, metabolic demand, and muscular stretch-shortening performance. Seventeen male soccer outfield players (16 ± 0 years, 66 ± 10 kg) performed three field shuttle-running tests with 15, 20, and 25-sec recoveries. In addition to specific shuttle test's variables, blood lactate concentration and vertical jump height were assessed. Resulting measures were highly reliable (intra-class correlation coefficient up to 0.86). 25-sec recovery improved test performance (-3% total time from 15-sec to 25-sec recovery), vertical jump height (+7% post-test height from 15-sec to 25-sec recovery), and decreased blood lactate accumulation (-33% post-test from 15-sec to 25-sec recovery). Study findings suggest that metabolic acidosis plays a role in worsening performance and fatigue development during the shuttle test. A 25-sec recovery duration maximized performance, containing metabolic-anaerobic power involvement and muscular stretch-shortening performance deterioration during a RSA test.

A program to evaluate a control system based on feedback of aerodynamic pressure differentials

NASA Technical Reports Server (NTRS)

Levy, D. W.; Finn, P.; Roskam, J.

1981-01-01

The use of aerodynamic pressure differentials to position a control surface is evaluated. The system is a differential pressure command loop, analogous to a position command loop, where the surface is commanded to move until a desired differential pressure across the surface is achieved. This type of control is more direct and accurate because it is the differential pressure which causes the control forces and moments. A frequency response test was performed in a low speed wind tunnel to measure the performance of the system. Both pressure and position feedback were tested. The pressure feedback performed as well as position feedback implying that the actuator, with a break frequency on the order of 10 Rad/sec, was the limiting component. Theoretical considerations indicate that aerodynamic lags will not appear below frequencies of 50 Rad/sec, or higher.

Effects on biological systems of reflected light from a satellite power system

NASA Technical Reports Server (NTRS)

White, M.

1981-01-01

Light reflection produced by the satellite power system and the possible effects of that light on the human eye, plants, and animals were studied. For the human eye, two cases of reflected light, might cause eye damage if viewed for too long. These cases are: (1) if, while in low Earth orbit, the orbital transfer vehicle is misaligned to reflect the Sun to Earth there exists a maximum safe fixation time for the naked eye of 42.4 secs; (2) reflection from the aluminum paint on the back of the orbital transfer vehicle, while in or near low Earth orbit, can be safely viewed by the naked eye for 129 sec. For plants and animals the intensity and timing of light are not a major problem. Ways for reducing and/or eliminating the irradiances are proposed.

The NASA/USAF arcjet research and technology program

NASA Technical Reports Server (NTRS)

Stone, James R.; Huston, Edward S.

1987-01-01

Direct current arcjets have the potential to provide specific impulses greater than 500 sec with storable propellants, and greater than 1000 sec with hydrogen. This level of performance can provide significant benefits for such applications as orbit transfer, station keeping, orbit change, and maneuvering. The simplicity of the arcjet system and its elements of commonality with state-of-the-art resistojet systems offer a relatively low risk transition to these enhanced levels of performance for low power (0.5 to 1.5 kW) station keeping applications. Arcjets at power levels of 10 to 30 kW are potentially applicable to orbit transfer missions. Furthermore, with the anticipated development of space nuclear power systems, arcjets at greater than 100 kW may become attractive. This paper describes the ongoing NASA/USAF program and describes major recent accomplishments.

Identification of secreted bacterial proteins by noncanonical amino acid tagging

PubMed Central

Mahdavi, Alborz; Szychowski, Janek; Ngo, John T.; Sweredoski, Michael J.; Graham, Robert L. J.; Hess, Sonja; Schneewind, Olaf; Mazmanian, Sarkis K.; Tirrell, David A.

2014-01-01

Pathogenic microbes have evolved complex secretion systems to deliver virulence factors into host cells. Identification of these factors is critical for understanding the infection process. We report a powerful and versatile approach to the selective labeling and identification of secreted pathogen proteins. Selective labeling of microbial proteins is accomplished via translational incorporation of azidonorleucine (Anl), a methionine surrogate that requires a mutant form of the methionyl-tRNA synthetase for activation. Secreted pathogen proteins containing Anl can be tagged by azide-alkyne cycloaddition and enriched by affinity purification. Application of the method to analysis of the type III secretion system of the human pathogen Yersinia enterocolitica enabled efficient identification of secreted proteins, identification of distinct secretion profiles for intracellular and extracellular bacteria, and determination of the order of substrate injection into host cells. This approach should be widely useful for the identification of virulence factors in microbial pathogens and the development of potential new targets for antimicrobial therapy. PMID:24347637

Interdependence of the Ypt/RabGAP Gyp5p and Gyl1p for recruitment to the sites of polarized growth.

PubMed

Chesneau, Laurent; Prigent, Magali; Boy-Marcotte, Emmanuelle; Daraspe, Jean; Fortier, Guillaume; Jacquet, Michel; Verbavatz, Jean-Marc; Cuif, Marie-Hélène

2008-04-01

Gyp5p and Gyl1p are two members of the Ypt/Rab guanosine triphosphatases-activating proteins involved in the control of polarized exocytosis in Saccharomyces cerevisiae. We had previously shown that Gyp5p and Gyl1p colocalize at the sites of polarized growth and belong to the same complex in subcellular fractions enriched in plasma membrane or secretory vesicles. Here, we investigate the interaction between Gyp5p and Gyl1p as well as the mechanism of their localization to the sites of polarized growth. We show that purified recombinant Gyp5p and Gyl1p interact directly in vitro. In vivo, both Gyp5p and Gyl1p are mutually required to concentrate at the sites of polarized growth. Moreover, the localization of Gyp5p and Gyl1p to the sites of polarized growth requires the formins Bni1p and Bnr1p and depends on actin cables. We show that, in a sec6-4 mutant, blocking secretion leads to coaccumulation of Gyp5p and Gyl1p, together with Sec4p. Electron microscopy experiments demonstrate that Gyp5p is associated with secretory vesicles. Altogether, our results indicate that both Gyp5p and Gyl1p access the sites of polarized growth by transport on secretory vesicles. Two polarisome components, Spa2p and Bud6p, are involved in maintaining Gyp5p and Gyl1p colocalized at the sites of polarized growth.

Substance P and vasoactive intestinal peptide degradation by mast cell tryptase and chymase.

PubMed

Caughey, G H; Leidig, F; Viro, N F; Nadel, J A

1988-01-01

The peptides substance P (SP) and vasoactive intestinal peptide (VIP) released from peptidergic neurons have potent effects on gland secretion and on smooth muscle tone. Because mast cells release proteases during degranulation, and are located in many of the same tissue microenvironments into which SP and VIP are released, we wished to examine whether mast cell proteases, by cleaving and thus inactivating these peptides, could modulate their effects. We used active site-titrated preparations of the two major neutral proteases of mast cell granules, tryptase and chymase, to determine the sites and rates of cleavage of SP and VIP. The proteases were purified from dog mastocytomas. Tryptase cleaved VIP rapidly at two sites with a kcat/Km of 2.2 X 10(5) sec-1 M-1, but had no effect on SP. Chymase cleaved both SP and VIP at primarily a single site with kcat/Km of 3.9 X 10(4) and 5.4 X 10(4) sec-1 M-1, respectively. Thus, these data show that mast cell proteases degrade SP and VIP. The differences in peptidase activity between tryptase and chymase suggest that the consequences of protease release could vary according to mast cell protease phenotype and location in various tissues and species. Tryptase, by cleaving the bronchodilator VIP but not the bronchoconstrictor SP, might promote bronchial hyper-responsiveness in asthma by decreasing the nonadrenergic neural inhibitory influence mediated by VIP. In skin and other tissues, chymase might interrupt axon reflex-mediated neurogenic inflammation by cleaving SP.

Traffic routing in a switched regenerative satellite. Volume 2, task 4: Review (executive summary)

NASA Astrophysics Data System (ADS)

Barberis, G.

1982-12-01

A communications satellite design for 1990's European use is proposed. Time plan assignment is discussed. The satellite system should be interfaced with the terrestrial telephone network at several nodes per nation (rather than at one node per nation as for ECS). A 64 Kbit/sec ISDN service and a specialized 2.048 Mbit/sec service on reservation basis are suggested. Wide use of the 12/14 and 20/30 GHz bands is advocated. A procedure to achieve a switching plan with high efficiency, taking into account all system constraints such as no bursts breaking and two transmission rates harmonization is proposed. Algorithms to be implemented are: the Hungarian method; branch and bound; the INSERT heuristic; and the HOLE heuristic.

Exploring the significance of structural hierarchy in material systems-A review

NASA Astrophysics Data System (ADS)

Pan, Ning

2014-06-01

Structural hierarchy and heterogeneity are inherent features in biological materials, but their significance in affecting the system behaviors is yet to be fully understood. In Sec. I, this article first identifies the major characteristics that manifest, or are resulted from, such hierarchy and heterogeneity in materials. Then in Sec. II, it presents several typical natural material systems including wood, bone, and others from animals to illustrate the proposed views. The paper also discusses a man-made smart material, textiles, to demonstrate that textiles are hierarchal, multifunctional, highly complex, and arguably the engineered material closest on a par with biological materials in complexity, and, more importantly, we can still learn quite a few new things from them in development of novel materials. In Sec. III, the paper summarizes several general approaches in developing a hierarchal material system at various scales, including structure thinning and splitting, laminating and layering, spatial and angular orientation, heterogenization and hybridization, and analyzes the advantages associated with them. It also stresses the adverse consequences once the existing structural hierarchy breaks down due to various mutations in biological systems. It discusses, in particular, the influences of moisture and air on material properties, given the near ubiquitousness of both air and water in materials. It next deals with in Sec. IV, some theoretical issues in material research including packing and ordering, the bi-modular mechanics, the behavior non-affinities due to disparity in hierarchal levels, the importance of system dimensionality in a hierarchal material system, and more philosophically, the issues of Nature's wisdom versus Intelligent Design. Section V then offers some concluding remarks, including a recap of the major issues covered in this article, and some general conclusions derived from the analyses and discussions. The main purpose of this paper is to make an effort to explore, identify, derive, or theorize some generic principles based on the existing results, not to offer another comprehensive review of current research activities in the fields for that there already exist some excellent ones. This paper examines the related topics with several approaches to not only reveal the underlying geometrical and physical mechanisms but also to emphasize the ways in which such mechanisms may be applied to developing engineered material systems with novel properties.

40 CFR 162.156 - General requirements.

Code of Federal Regulations, 2010 CFR

2010-07-01

... thereunder: (i) Sec. 12(a)(1) (A) through (E), in accordance with: (A) Sec. 2(q)(1) (A) through (G). (B) Sec... registered by the Administrator in accordance with sec. 7 of FIFRA and 40 CFR part 167. (c) Books and records...

7 CFR 235.4 - Allocation of funds to States.

Code of Federal Regulations, 2010 CFR

2010-01-01

.... (Sec. 14, Pub. L. 95-166, 91 Stat. 1338 (42 U.S.C. 1776); sec. 7, Pub. L. 95-627, 92 Stat. 3621 (42 U.S.C. 1776); sec. 7(a), Pub. L. 95-627, 92 Stat. 3622 (42 U.S.C. 1751); Pub. L. 96-499, secs. 201 and..., 1759a, 1774 and 1776); E.O. 12372 (July 14, 1982, 47 FR 30959); sec. 401(b) Intergovernmental...

7 CFR 235.4 - Allocation of funds to States.

Code of Federal Regulations, 2012 CFR

2012-01-01

.... (Sec. 14, Pub. L. 95-166, 91 Stat. 1338 (42 U.S.C. 1776); sec. 7, Pub. L. 95-627, 92 Stat. 3621 (42 U.S.C. 1776); sec. 7(a), Pub. L. 95-627, 92 Stat. 3622 (42 U.S.C. 1751); Pub. L. 96-499, secs. 201 and..., 1759a, 1774 and 1776); E.O. 12372 (July 14, 1982, 47 FR 30959); sec. 401(b) Intergovernmental...

7 CFR 235.4 - Allocation of funds to States.

Code of Federal Regulations, 2013 CFR

2013-01-01

.... (Sec. 14, Pub. L. 95-166, 91 Stat. 1338 (42 U.S.C. 1776); sec. 7, Pub. L. 95-627, 92 Stat. 3621 (42 U.S.C. 1776); sec. 7(a), Pub. L. 95-627, 92 Stat. 3622 (42 U.S.C. 1751); Pub. L. 96-499, secs. 201 and..., 1759a, 1774 and 1776); E.O. 12372 (July 14, 1982, 47 FR 30959); sec. 401(b) Intergovernmental...

7 CFR 235.4 - Allocation of funds to States.

Code of Federal Regulations, 2014 CFR

2014-01-01

.... (Sec. 14, Pub. L. 95-166, 91 Stat. 1338 (42 U.S.C. 1776); sec. 7, Pub. L. 95-627, 92 Stat. 3621 (42 U.S.C. 1776); sec. 7(a), Pub. L. 95-627, 92 Stat. 3622 (42 U.S.C. 1751); Pub. L. 96-499, secs. 201 and..., 1759a, 1774 and 1776); E.O. 12372 (July 14, 1982, 47 FR 30959); sec. 401(b) Intergovernmental...

7 CFR 235.4 - Allocation of funds to States.

Code of Federal Regulations, 2011 CFR

2011-01-01

.... (Sec. 14, Pub. L. 95-166, 91 Stat. 1338 (42 U.S.C. 1776); sec. 7, Pub. L. 95-627, 92 Stat. 3621 (42 U.S.C. 1776); sec. 7(a), Pub. L. 95-627, 92 Stat. 3622 (42 U.S.C. 1751); Pub. L. 96-499, secs. 201 and..., 1759a, 1774 and 1776); E.O. 12372 (July 14, 1982, 47 FR 30959); sec. 401(b) Intergovernmental...

Activation of Rab GTPase Sec4 by its GEF Sec2 is required for prospore membrane formation during sporulation in yeast Saccharomyces cerevisiae.

PubMed

Suda, Yasuyuki; Tachikawa, Hiroyuki; Inoue, Ichiro; Kurita, Tomokazu; Saito, Chieko; Kurokawa, Kazuo; Nakano, Akihiko; Irie, Kenji

2018-02-01

Sec2 activates Sec4 Rab GTPase as a guanine nucleotide exchange factor for the recruitment of downstream effectors to facilitate tethering and fusion of post-Golgi vesicles at the plasma membrane. During the meiosis and sporulation of budding yeast, post-Golgi vesicles are transported to and fused at the spindle pole body (SPB) to form a de novo membrane, called the prospore membrane. Previous studies have revealed the role of the SPB outer surface called the meiotic outer plaque (MOP) in docking and fusion of post-Golgi vesicles. However, the upstream molecular machinery for post-Golgi vesicular fusion that facilitates prospore membrane formation remains enigmatic. Here, we demonstrate that the GTP exchange factor for Sec4, Sec2, participates in the formation of the prospore membrane. A conditional mutant in which the SEC2 expression is shut off during sporulation showed sporulation defects. Inactivation of Sec2 caused Sec4 targeting defects along the prospore membranes, thereby causing insufficient targeting of downstream effectors and cargo proteins to the prospore membrane. These results suggest that the activation of Sec4 by Sec2 is required for the efficient supply of post-Golgi vesicles to the prospore membrane and thus for prospore membrane formation/extension and subsequent deposition of spore wall materials. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

SEC16A is a RAB10 effector required for insulin-stimulated GLUT4 trafficking in adipocytes

PubMed Central

Bruno, Joanne; Chaudhary, Natasha; Iaea, David

2016-01-01

RAB10 is a regulator of insulin-stimulated translocation of the GLUT4 glucose transporter to the plasma membrane (PM) of adipocytes, which is essential for whole-body glucose homeostasis. We establish SEC16A as a novel RAB10 effector in this process. Colocalization of SEC16A with RAB10 is augmented by insulin stimulation, and SEC16A knockdown attenuates insulin-induced GLUT4 translocation, phenocopying RAB10 knockdown. We show that SEC16A and RAB10 promote insulin-stimulated mobilization of GLUT4 from a perinuclear recycling endosome/TGN compartment. We propose RAB10–SEC16A functions to accelerate formation of the vesicles that ferry GLUT4 to the PM during insulin stimulation. Because GLUT4 continually cycles between the PM and intracellular compartments, the maintenance of elevated cell-surface GLUT4 in the presence of insulin requires accelerated biogenesis of the specialized GLUT4 transport vesicles. The function of SEC16A in GLUT4 trafficking is independent of its previously characterized activity in ER exit site formation and therefore independent of canonical COPII-coated vesicle function. However, our data support a role for SEC23A, but not the other COPII components SEC13, SEC23B, and SEC31, in the insulin stimulation of GLUT4 trafficking, suggesting that vesicles derived from subcomplexes of COPII coat proteins have a role in the specialized trafficking of GLUT4. PMID:27354378

76 FR 47516 - Personnel Management in Agencies

Federal Register 2010, 2011, 2012, 2013, 2014

2011-08-05

... management, in accordance with merit system principles. Sec. 250.205 Systems and standards. The five human... effective human capital management in accordance with the merit system principles and in compliance with... OFFICE OF PERSONNEL MANAGEMENT 5 CFR PART 250 RIN 3206-AL98 Personnel Management in Agencies...

Identification of isopropylbiphenyl, alkyl diphenylmethanes, diisopropylnaphthalene, linear alkyl benzenes and other polychlorinated biphenyl replacement compounds in effluents, sediments and fish in the Fox River System, Wisconsin

USGS Publications Warehouse

Peterman, Paul H.; Delfino, Joseph J.

1990-01-01

Five polychlorinated biphenyl replacement dye solvents and a diluent present in carbonless copy paper were identified by gas chromatography/mass spectrometry in the following matrices: effluents from a de-inking–recycling paper mill and a municipal wastewater treatment plant receiving wastewaters from a carbonless copy paper manufacturing plant; sediments; and fish collected near both discharges in the Fox River System, Wisconsin. An isopropylbiphenyl dye solvent mixture included mono-, di- and triisopropylbiphenyls. Also identified were two dye solvent mixtures marketed under the trade name Santosol. Santosol 100 comprised ethyl-diphenylmethanes (DPMs), benzyl-ethyl-DPMs, and dibenzyl-ethyl-DPMs. Similarly, Santosol 150 comprised dimethyl-DPMs, benzyl-dimethyl-DPMs, and dibenzyl-dimethyl-DPMs. Diisopropylnaphthalenes, widely used as a dye solvent in Japan, were identified for the first time in the US environment. sec-Butylbiphenyls and di-sec-butylbiphenyls, likely constituents of a sec-butylbiphenyl dye solvent mixture, were tentatively identified. Linear alkyl benzenes (C10 to C13-LABs) constituted the Alkylate 215 diluent mixture. Although known to occur as minor constituents in linear alkyl sulfonate detergents, LAB residues have not been previously attributed to commercial use of LABs.

One year quality of life measured with SEC-QoL in levonorgestrel 52 mg IUS users.

PubMed

Cristobal, Ignacio; Lete, Luis Ignacio; Viuda, Esther de la; Perulero, Nuria; Arbat, Agnes; Canals, Ignasi

2016-04-01

The present study aims to prospectively evaluate quality of life (QoL) of women using 52-mg levonorgestrel intrauterine system (LNG-IUS) for contraception determined through the Sociedad Española de Contracepción (Spanish contraception Society) (SEC)-QoL, a questionnaire specifically designed to assess the impact of contraceptive methods on QoL of fertile women. We conducted a prospective observational multicenter study of 201 reproductive age women who initiated the LNG-IUS for contraception. Sociodemographic and clinical data were collected at baseline and 12 months afterwards. Participants filled in the SEC-QoL questionnaire at both visits. SEQ-QoL scores range from 0 (worst QoL) to 100 (best QoL). Participants claimed an increased QoL 12 months after insertion in all five dimensions of SEC-QoL due to its high contraceptive efficacy and its capability to reduce other menstrual symptoms (e.g., heavy menstrual bleeding or dysmenorrhoea), overall exerting a positive impact on user's satisfaction. SEC-QoL general overall score went from a mean (S.D.) score of 46.3 (17.3) at baseline to 72.2 (14.8) 12 months afterwards (p<.001). Overall, 94.6% of women claimed having found additional benefits other than contraception. No pregnancies were reported during the 12 months of study duration, and only 14 women discontinued use of LNG-IUS (only two of them due to an adverse event), representing a continuation rate of 93%. Women using LNG-IUS for contraception have an increased QoL after 12 months of use, demonstrated by the increased score in all dimensions of the SEC-QoL questionnaire. The present study prospectively evaluated QoL of women using LNG-IUS for contraception through the SEC-QoL questionnaire. Participants claimed increased QoL 12 months afterwards, implying that women using LNG-IUS for contraception in usual clinical practise also benefit from the reduction of period-related symptoms, overall leading to very low discontinuation rates. Copyright © 2016 Elsevier Inc. All rights reserved.

Single-mode 850-nm vertical-cavity surface-emitting lasers with Zn-diffusion and oxide-relief apertures for > 50 Gbit/sec OOK and 4-PAM transmission

NASA Astrophysics Data System (ADS)

Shi, Jin-Wei; Wei, Chia-Chien; Chen, Jyehong; Ledentsov, N. N.; Yang, Ying-Jay

2017-02-01

Vertical-cavity surface-emitting lasers (VCSELs) has become the most important light source in the booming market of short-reach (< 300 meters) optical interconnect (OI). The next generation OI has been targeted at 56 Gbit/sec data rate per channel (CEI-56G) with the total data rate up to 400 Gbit/sec. However, the serious modal dispersion of multi-mode fiber (MMF), limited speed of VCSEL, and its high resistance (> 150 Ω) seriously limits the >50 Gbit/sec linking distance (< 10 m) by using only on-off keying (OOK) modulation scheme without any signal processing techniques. In contrast to OOK, 4-PAM modulation format is attractive for >50 Gbit/sec transmission due to that it can save one-half of the required bandwidth. Nevertheless, a 4.7 dB optical power penalty and the linearity of transmitter would become issues in the 4-PAM linking performance. Besides, in the modern OI system, the optics transreceiver module must be packaged as close as possible with the integrated circuits (ICs). The heat generated from ICs will become an issue in speed of VSCEL. Here, we review our recent work about 850 nm VCSEL, which has unique Zn-diffusion/oxide-relief apertures and special p- doping active layer with strong wavelength detuning to further enhance its modulation speed and high-temperature (85°C) performances. Single-mode (SM) devices with high-speed ( 26 GHz), reasonable resistance ( 70 Ω) and moderate output power ( 1.5 mW) can be achieved. Error-free 54 Gbit/sec OOK transmission through 1km MMF has been realized by using such SM device with signal processing techniques. Besides, the volterra nonlinear equalizer has been applied in our 4-PAM 64 Gbit/sec transmission through 2-km OM4 MMF, which significantly enhance the linearity of device and outperforms fed forward equalization (FFE) technique. Record high bit-rate distance product of 128.km is confirmed for optical-interconnect applications.

A Pseudo-Atomic Model of the COPII Cage Obtained from CryoEM and Mass Spectrometry Analyses

PubMed Central

Noble, Alex J.; Zhang, Qian; O’Donnell, Jason; Hariri, Hanaa; Bhattacharya, Nilakshee; Marshall, Alan G.

2012-01-01

COPII vesicles transport proteins from the ER to the Golgi apparatus. Previous cryoEM structures of the COPII cage lacked the resolution necessary to determine the residues of Sec13 and Sec31 that mediate assembly and flexibility of the COPII cage. Here we present a 12Å-resolution structure of the COPII cage, where the tertiary structure of Sec13 and Sec31 is clearly identifiable. We employ this structure and a homology model of the Sec13-Sec31 complex to create a reliable pseudo-atomic model of the COPII cage. We combined this model with hydrogen/deuterium exchange mass spectrometry analysis to characterize four distinct contact regions at the vertices of the COPII cage. Furthermore, we found that the 2-fold symmetry of the Sec31 dimeric region of Sec13-31 is broken on cage formation, and that the resulting hinge is essential to form the proper edge geometry in COPII cages. PMID:23262493

Agonal sequences in eight filmed hangings: analysis of respiratory and movement responses to asphyxia by hanging.

PubMed

Sauvageau, Anny; LaHarpe, Romano; Geberth, Vernon J

2010-09-01

It has been proposed that filmed hangings may hold the key to a better understanding of human asphyxia, and The Working Group on Human Asphyxia was formed to systematically review and compare these video recordings. This study analyzed eight filmed hangings. Considering time 0 to represent the onset of the final hanging, rapid loss of consciousness was observed (at 8-18 sec), closely followed by convulsions (at 10-19 sec). A complex pattern of decerebrate rigidity and decorticate rigidity then followed. Between 1 min 38 sec and 2 min 15 sec, muscle tone seemed to be lost, the body becoming progressively flaccid. From then on, isolated body movements were observed from time to time, the last one occurring between 1 min 2 sec and 7 min 31 sec. As for the respiratory responses, all cases presented deep rhythmic abdominal respiratory movements (last one between 1 min 2 sec and 2 min 5 sec). © 2010 American Academy of Forensic Sciences.

Regulation of the protein-conducting channel by a bound ribosome

PubMed Central

Gumbart, James; Trabuco, Leonardo G.; Schreiner, Eduard; Villa, Elizabeth; Schulten, Klaus

2009-01-01

Summary During protein synthesis, it is often necessary for the ribosome to form a complex with a membrane-bound channel, the SecY/Sec61 complex, in order to translocate nascent proteins across a cellular membrane. Structural data on the ribosome-channel complex are currently limited to low-resolution cryo-electron microscopy maps, including one showing a bacterial ribosome bound to a monomeric SecY complex. Using that map along with available atomic-level models of the ribosome and SecY, we have determined, through molecular dynamics flexible fitting (MDFF), an atomic-resolution model of the ribosome-channel complex. We characterized computationally the sites of ribosome-SecY interaction within the complex and determined the effect of ribosome binding on the SecY channel. We also constructed a model of a ribosome in complex with a SecY dimer by adding a second copy of SecY to the MDFF-derived model. The study involved 2.7-million-atom simulations over altogether nearly 50 ns. PMID:19913480

IcmS-dependent translocation of SdeA into macrophages by the Legionella pneumophila type IV secretion system.

PubMed

Bardill, J Patrick; Miller, Jennifer L; Vogel, Joseph P

2005-04-01

Legionella pneumophila replicates inside alveolar macrophages and causes an acute, potentially fatal pneumonia called Legionnaires' disease. The ability of this bacterium to grow inside of macrophages is dependent on the presence of a functional dot/icm type IV secretion system (T4SS). Proteins secreted by the Dot/Icm T4SS are presumed to alter the host endocytic pathway, allowing L. pneumophila to establish a replicative niche within the host cell. Here we show that a member of the SidE family of proteins interacts with IcmS and is required for full virulence in the protozoan host Acanthamoeba castellanii. Using immunofluorescence microscopy and adenylate cyclase fusions, we show that SdeA is secreted into host cells by L. pneumophila in an IcmS-dependent manner. The SidE-like proteins are secreted very early during macrophage infection, suggesting that they are important in the initial formation of the replicative phagosome. Secreted SidE family members show a similar localization to other Dot/Icm substrates, specifically, to the poles of the replicative phagosome. This common localization of secreted substrates of the Dot/Icm system may indicate the formation of a multiprotein complex on the cytoplasmic face of the replicative phagosome.

Central Nervous System Neuropeptide Y Signaling Modulates VLDL Triglyceride Secretion

PubMed Central

Stafford, John M.; Yu, Fang; Printz, Richard; Hasty, Alyssa H.; Swift, Larry L.; Niswender, Kevin D.

2014-01-01

OBJECTIVE Elevated triglyceride (TG) is the major plasma lipid abnormality in obese and diabetic patients and contributes to cardiovascular morbidity in these disorders. We sought to identify novel mechanisms leading to hypertriglyceridemia. Resistance to negative feedback signals from adipose tissue in key central nervous system (CNS) energy homeostatic circuits contributes to the development of obesity. Because triglycerides both represent the largest energy depot in the body and are elevated in both the plasma and adipose in obesity and diabetes, we hypothesized that the same neural circuits that regulate energy balance also regulate the secretion of TGs into plasma. RESEARCH DESIGN AND METHODS In normal fasting rats, the TG secretion rate was estimated by serial blood sampling after intravascular tyloxapol pretreatment. Neuropeptide Y (NPY) signaling in the CNS was modulated by intracerebroventricular injection of NPY, receptor antagonist, and receptor agonist. RESULTS A single intracerebroventricular injection of NPY increased TG secretion by 2.5-fold in the absence of food intake, and this was determined to be VLDL by fast performance liquid chromatography (FPLC). This effect was recapitulated by activating NPY signaling in downstream neurons with an NPY-Y5 receptor agonist. An NPY-Y1 receptor antagonist decreased the elevated TGs in the form of VLDL secretion rate by 50% compared with vehicle. Increased TG secretion was due to increased secretion of VLDL particles, rather than secretion of larger particles, because apolipoprotein B100 was elevated in FPLC fractions corresponding to VLDL. CONCLUSIONS We find that a key neuropeptide system involved in energy homeostasis in the CNS exerts control over VLDL-TG secretion into the bloodstream. PMID:18332095

The role of intrinsic disorder and dynamics in the assembly and function of the type II secretion system.

PubMed

Gu, Shuang; Shevchik, Vladimir E; Shaw, Rosie; Pickersgill, Richard W; Garnett, James A

2017-10-01

Many Gram-negative commensal and pathogenic bacteria use a type II secretion system (T2SS) to transport proteins out of the cell. These exported proteins or substrates play a major role in toxin delivery, maintaining biofilms, replication in the host and subversion of host immune responses to infection. We review the current structural and functional work on this system and argue that intrinsically disordered regions and protein dynamics are central for assembly, exo-protein recognition, and secretion competence of the T2SS. The central role of intrinsic disorder-order transitions in these processes may be a particular feature of type II secretion. Copyright © 2017 Elsevier B.V. All rights reserved.

77 FR 55153 - Magnuson-Stevens Act Provisions; Fisheries Off West Coast States; Pacific Coast Groundfish...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-09-07

... regulations erroneously state that an operating system such as ``Windows 2007'' may be used; there is no such operating system so this system is removed. Corrections to Regulations Implemented as Part of the 2011-2012... follows: Sec. 660.15 Equipment requirements. * * * * * (d) * * * (1) * * * (v) Operating system: Microsoft...

A gatekeeper chaperone complex directs translocator secretion during Type Three Secretion

DOE PAGES

Archuleta, Tara L.; Spiller, Benjamin W.; Kubori, Tomoko

2014-11-06

Many Gram-negative bacteria use Type Three Secretion Systems (T3SS) to deliver effector proteins into host cells. These protein delivery machines are composed of cytosolic components that recognize substrates and generate the force needed for translocation, the secretion conduit, formed by a needle complex and associated membrane spanning basal body, and translocators that form the pore in the target cell. A defined order of secretion in which needle component proteins are secreted first, followed by translocators, and finally effectors, is necessary for this system to be effective. While the secreted effectors vary significantly between organisms, the ~20 individual protein components thatmore » form the T3SS are conserved in many pathogenic bacteria. One such conserved protein, referred to as either a plug or gatekeeper, is necessary to prevent unregulated effector release and to allow efficient translocator secretion. The mechanism by which translocator secretion is promoted while effector release is inhibited by gatekeepers is unknown. We present the structure of the Chlamydial gatekeeper, CopN, bound to a translocator-specific chaperone. The structure identifies a previously unknown interface between gatekeepers and translocator chaperones and reveals that in the gatekeeper-chaperone complex the canonical translocator-binding groove is free to bind translocators. Thus, structure-based mutagenesis of the homologous complex in Shigella reveals that the gatekeeper-chaperone-translocator complex is essential for translocator secretion and for the ordered secretion of translocators prior to effectors.« less

Characterization of Nops, nodulation outer proteins, secreted via the type III secretion system of NGR234.

PubMed

Marie, Corinne; Deakin, William J; Viprey, Virginie; Kopciñska, Joanna; Golinowski, Wladyslaw; Krishnan, Hari B; Perret, Xavier; Broughton, William J

2003-09-01

The nitrogen-fixing symbiotic bacterium Rhizobium species NGR234 secretes, via a type III secretion system (TTSS), proteins called Nops (nodulation outer proteins). Abolition of TTSS-dependent protein secretion has either no effect or leads to a change in the number of nodules on selected plants. More dramatically, Nops impair nodule development on Crotalaria juncea roots, resulting in the formation of nonfixing pseudonodules. A double mutation of nopX and nopL, which code for two previously identified secreted proteins, leads to a phenotype on Pachyrhizus tuberosus differing from that of a mutant in which the TTSS is not functional. Use of antibodies and a modification of the purification protocol revealed that NGR234 secretes additional proteins in a TTSS-dependent manner. One of them was identified as NopA, a small 7-kDa protein. Single mutations in nopX and nopL were also generated to assess the involvement of each Nop in protein secretion and nodule formation. Mutation of nopX had little effect on NopL and NopA secretion but greatly affected the interaction of NGR234 with many plant hosts tested. NopL was not necessary for the secretion of any Nops but was required for efficient nodulation of some plant species. NopL may thus act as an effector protein whose recognition is dependent upon the hosts' genetic background.

Characterization of Shikonin Derivative Secretion in Lithospermum erythrorhizon Hairy Roots as a Model of Lipid-Soluble Metabolite Secretion from Plants

PubMed Central

Tatsumi, Kanade; Yano, Mariko; Kaminade, Kenta; Sugiyama, Akifumi; Sato, Mayuko; Toyooka, Kiminori; Aoyama, Takashi; Sato, Fumihiko; Yazaki, Kazufumi

2016-01-01

Shikonin derivatives are specialized lipophilic metabolites, secreted in abundant amounts from the root epidermal cells of Lithospermum erythrorhizon. Because they have anti-microbial activities, these compounds, which are derivatives of red naphthoquinone, are thought to serve as a chemical barrier for plant roots. The mechanism by which they are secreted from cells is, however, largely unknown. The shikonin production system in L. erythrorhizon is an excellent model for studying the mechanism by which lipophilic compounds are secreted from plant cells, because of the abundant amounts of these compounds produced by L. erythrorhizon, the 0 to 100% inducibility of their production, the light-specific inhibition of production, and the visibility of these products as red pigments. To date, many factors regulating shikonin biosynthesis have been identified, but no mechanism that regulates shikonin secretion without inhibiting biosynthesis has been detected. This study showed that inhibitors of membrane traffic strongly inhibit shikonin secretion without inhibiting shikonin production, suggesting that the secretion of shikonin derivatives into the apoplast utilizes pathways common to the ADP-ribosylation factor/guanine nucleotide exchange factor (ARF/GEF) system and actin filament polymerization, at least in part. These findings provide clues about the machinery involved in secreting lipid-soluble metabolites from cells. PMID:27507975

Characterization of Shikonin Derivative Secretion in Lithospermum erythrorhizon Hairy Roots as a Model of Lipid-Soluble Metabolite Secretion from Plants.

PubMed

Tatsumi, Kanade; Yano, Mariko; Kaminade, Kenta; Sugiyama, Akifumi; Sato, Mayuko; Toyooka, Kiminori; Aoyama, Takashi; Sato, Fumihiko; Yazaki, Kazufumi

2016-01-01

Shikonin derivatives are specialized lipophilic metabolites, secreted in abundant amounts from the root epidermal cells of Lithospermum erythrorhizon. Because they have anti-microbial activities, these compounds, which are derivatives of red naphthoquinone, are thought to serve as a chemical barrier for plant roots. The mechanism by which they are secreted from cells is, however, largely unknown. The shikonin production system in L. erythrorhizon is an excellent model for studying the mechanism by which lipophilic compounds are secreted from plant cells, because of the abundant amounts of these compounds produced by L. erythrorhizon, the 0 to 100% inducibility of their production, the light-specific inhibition of production, and the visibility of these products as red pigments. To date, many factors regulating shikonin biosynthesis have been identified, but no mechanism that regulates shikonin secretion without inhibiting biosynthesis has been detected. This study showed that inhibitors of membrane traffic strongly inhibit shikonin secretion without inhibiting shikonin production, suggesting that the secretion of shikonin derivatives into the apoplast utilizes pathways common to the ADP-ribosylation factor/guanine nucleotide exchange factor (ARF/GEF) system and actin filament polymerization, at least in part. These findings provide clues about the machinery involved in secreting lipid-soluble metabolites from cells.

The importance of connections between the cell wall integrity pathway and the unfolded protein response in filamentous fungi.

PubMed

Malavazi, Iran; Goldman, Gustavo Henrique; Brown, Neil Andrew

2014-11-01

In the external environment, or within a host organism, filamentous fungi experience sudden changes in nutrient availability, osmolality, pH, temperature and the exposure to toxic compounds. The fungal cell wall represents the first line of defense, while also performing essential roles in morphology, development and virulence. A polarized secretion system is paramount for cell wall biosynthesis, filamentous growth, nutrient acquisition and interactions with the environment. The unique ability of filamentous fungi to secrete has resulted in their industrial adoption as fungal cell factories. Protein maturation and secretion commences in the endoplasmic reticulum (ER). The unfolded protein response (UPR) maintains ER functionality during exposure to secretion and cell wall stress. UPR, therefore, influences secretion and cell wall homeostasis, which in turn impacts upon numerous fungal traits important to pathogenesis and biotechnology. Subsequently, this review describes the relevance of the cell wall and UPR systems to filamentous fungal pathogens or industrial microbes and then highlights interconnections between the two systems. Ultimately, the possible biotechnological applications of an enhanced understanding of such regulatory systems in combating fungal disease, or the removal of natural bottlenecks in protein secretion in an industrial setting, are discussed. © The Author 2014. Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oup.com.

Inhibition of selenocysteine tRNA[Ser]Sec aminoacylation provides evidence that aminoacylation is required for regulatory methylation of this tRNA

PubMed Central

Kim, Jin Young; Carlson, Bradley A.; Xu, Xue-Ming; Zeng, Yu; Chen, Shawn; Gladyshev, Vadim N.; Lee, Byeong Jae; Hatfield, Dolph L.

2011-01-01

There are two isoforms of selenocysteine (Sec) tRNA[Ser]Sec that differ by a single methyl group, Um34. The non-Um34 isoform supports the synthesis of a subclass of selenoproteins, designated housekeeping, while the Um34 isoform supports the expression of another subclass, designated stress-related selenoproteins. Herein, we investigated the relationship between tRNA[Ser]Sec aminoacylation and Um34 synthesis which is the last step in the maturation of this tRNA. Mutation of the discriminator base at position 73 in tRNA[Ser]Sec dramatically reduced aminoacylation with serine, as did an inhibitor of seryl-tRNA synthetase, SB-217452. Although both the mutation and the inhibitor prevented Um34 synthesis, neither precluded the synthesis of any other of the known base modifications on tRNA[Ser]Sec following microinjection and incubation of the mutant tRNA[Ser]Sec transcript, or the wild type transcript along with inhibitor, in Xenopus oocytes. The data demonstrate that Sec tRNA[Ser]Sec must be aminoacylated for Um34 addition. The fact that selenium is required for Um34 methylation suggests that Sec must be attached to its tRNA for Um34 methylation. This would explain why selenium is essential for the function of Um34 methylase and provides further insights into the hierarchy of selenoprotein expression. PMID:21624347

Identification of the DotL coupling protein subcomplex of the Legionella Dot/Icm type IV secretion system.

PubMed

Vincent, Carr D; Friedman, Jonathan R; Jeong, Kwang Cheol; Sutherland, Molly C; Vogel, Joseph P

2012-07-01

Legionella pneumophila, the causative agent of Legionnaires' disease, survives in macrophages by altering the endocytic pathway of its host cell. To accomplish this, the bacterium utilizes a type IVB secretion system to deliver effector molecules into the host cell cytoplasm. In a previous report, we performed an extensive characterization of the L. pneumophila type IVB secretion system that resulted in the identification of a critical five-protein subcomplex that forms the core of the secretion apparatus. Here we describe a second Dot/Icm protein subassembly composed of the type IV coupling protein DotL, the apparatus proteins DotM and DotN, and the secretion adaptor proteins IcmS and IcmW. In the absence of IcmS or IcmW, DotL becomes destabilized at the transition from the exponential to stationary phases of growth, concurrent with the expression of many secreted substrates. Loss of DotL is dependent on ClpA, a regulator of the cytoplasmic protease ClpP. The resulting decreased levels of DotL in the icmS and icmW mutants exacerbates the intracellular defects of these strains and can be partially suppressed by overproduction of DotL. Thus, in addition to their role as chaperones for Legionella type IV secretion system substrates, IcmS and IcmW perform a second function as part of the Dot/Icm type IV coupling protein subcomplex. © 2012 Blackwell Publishing Ltd.

Transposon-disruption of a maize nuclear gene, tha1, encoding a chloroplast SecA homologue: in vivo role of cp-SecA in thylakoid protein targeting.

PubMed

Voelker, R; Mendel-Hartvig, J; Barkan, A

1997-02-01

A nuclear mutant of maize, tha1, which exhibited defects in the translocation of proteins across the thylakoid membrane, was described previously. A transposon insertion at the tha1 locus facilitated the cloning of portions of the tha1 gene. Strong sequence similarity with secA genes from bacteria, pea and spinach indicates that tha1 encodes a SecA homologue (cp-SecA). The tha1-ref allele is either null or nearly so, in that tha1 mRNA is undetectable in mutant leaves and cp-SecA accumulation is reduced > or = 40-fold. These results, in conjunction with the mutant phenotype described previously, demonstrate that cp-SecA functions in vivo to facilitate the translocation of OEC33, PSI-F and plastocyanin but does not function in the translocation of OEC23 and OEC16. Our results confirm predictions for cp-SecA function made from the results of in vitro experiments and establish several new functions for cp-SecA, including roles in the targeting of a chloroplast-encoded protein, cytochrome f, and in protein targeting in the etioplast, a nonphotosynthetic plastid type. Our finding that the accumulation of properly targeted plastocyanin and cytochrome f in tha1-ref thylakoid membranes is reduced only a few-fold despite the near or complete absence of cp-SecA suggests that cp-SecA facilitates but is not essential in vivo for their translocation across the membrane.

Association of early- and adult-life socioeconomic circumstances with muscle strength in older age.

PubMed

Cheval, Boris; Boisgontier, Matthieu P; Orsholits, Dan; Sieber, Stefan; Guessous, Idris; Gabriel, Rainer; Stringhini, Silvia; Blane, David; van der Linden, Bernadette W A; Kliegel, Matthias; Burton-Jeangros, Claudine; Courvoisier, Delphine S; Cullati, Stéphane

2018-05-01

socioeconomic circumstances (SEC) during a person's lifespan influence a wide range of health outcomes. However, solid evidence of the association of early- and adult-life SEC with health trajectories in ageing is still lacking. This study assessed whether early-life SEC are associated with muscle strength in later life-a biomarker of health-and whether this relationship is caused by adult-life SEC and health behaviours. we used data from the Survey of Health Ageing and Retirement in Europe, a 12-year population-based cohort study with repeated measurement in six waves (2004-15) and retrospective collection of life-course data. Participants' grip strength was assessed by using a handheld dynamometer. Confounder-adjusted logistic mixed-effect models were used to examine the associations of early- and adult-life SEC with the risk of low muscle strength (LMS) in older age. a total of 24,179 participants (96,375 observations) aged 50-96 living in 14 European countries were included in the analyses. Risk of LMS was increased with disadvantaged relative to advantaged early-life SEC. The association between risk of LMS and disadvantaged early-life SEC gradually decreased when adjusting for adult-life SEC for both sexes and with unhealthy behaviours for women. After adjusting for these factors, all associations between risk of LMS and early-life SEC remained significant for women. early-life SEC are associated with muscle strength after adjusting for adult-life SEC and behavioural lifestyle factors, especially in women, which suggests that early life may represent a sensitive period for future health.

76 FR 41993 - Customs and Border Protection Officer Retirement

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Characterization of Native Protein Complexes and Protein Isoform Variation Using Size-fractionation-based Quantitative Proteomics*

PubMed Central

Kirkwood, Kathryn J.; Ahmad, Yasmeen; Larance, Mark; Lamond, Angus I.

2013-01-01

Proteins form a diverse array of complexes that mediate cellular function and regulation. A largely unexplored feature of such protein complexes is the selective participation of specific protein isoforms and/or post-translationally modified forms. In this study, we combined native size-exclusion chromatography (SEC) with high-throughput proteomic analysis to characterize soluble protein complexes isolated from human osteosarcoma (U2OS) cells. Using this approach, we have identified over 71,500 peptides and 1,600 phosphosites, corresponding to over 8,000 proteins, distributed across 40 SEC fractions. This represents >50% of the predicted U2OS cell proteome, identified with a mean peptide sequence coverage of 27% per protein. Three biological replicates were performed, allowing statistical evaluation of the data and demonstrating a high degree of reproducibility in the SEC fractionation procedure. Specific proteins were detected interacting with multiple independent complexes, as typified by the separation of distinct complexes for the MRFAP1-MORF4L1-MRGBP interaction network. The data also revealed protein isoforms and post-translational modifications that selectively associated with distinct subsets of protein complexes. Surprisingly, there was clear enrichment for specific Gene Ontology terms associated with differential size classes of protein complexes. This study demonstrates that combined SEC/MS analysis can be used for the system-wide annotation of protein complexes and to predict potential isoform-specific interactions. All of these SEC data on the native separation of protein complexes have been integrated within the Encyclopedia of Proteome Dynamics, an online, multidimensional data-sharing resource available to the community. PMID:24043423

Characterization of native protein complexes and protein isoform variation using size-fractionation-based quantitative proteomics.

PubMed

Kirkwood, Kathryn J; Ahmad, Yasmeen; Larance, Mark; Lamond, Angus I

2013-12-01

Proteins form a diverse array of complexes that mediate cellular function and regulation. A largely unexplored feature of such protein complexes is the selective participation of specific protein isoforms and/or post-translationally modified forms. In this study, we combined native size-exclusion chromatography (SEC) with high-throughput proteomic analysis to characterize soluble protein complexes isolated from human osteosarcoma (U2OS) cells. Using this approach, we have identified over 71,500 peptides and 1,600 phosphosites, corresponding to over 8,000 proteins, distributed across 40 SEC fractions. This represents >50% of the predicted U2OS cell proteome, identified with a mean peptide sequence coverage of 27% per protein. Three biological replicates were performed, allowing statistical evaluation of the data and demonstrating a high degree of reproducibility in the SEC fractionation procedure. Specific proteins were detected interacting with multiple independent complexes, as typified by the separation of distinct complexes for the MRFAP1-MORF4L1-MRGBP interaction network. The data also revealed protein isoforms and post-translational modifications that selectively associated with distinct subsets of protein complexes. Surprisingly, there was clear enrichment for specific Gene Ontology terms associated with differential size classes of protein complexes. This study demonstrates that combined SEC/MS analysis can be used for the system-wide annotation of protein complexes and to predict potential isoform-specific interactions. All of these SEC data on the native separation of protein complexes have been integrated within the Encyclopedia of Proteome Dynamics, an online, multidimensional data-sharing resource available to the community.

Kinetics of cycle length dependence of ventricular repolarization: effect of autonomic blockade

NASA Technical Reports Server (NTRS)

Raeder, E. A.; Albrecht, P.; Perrott, M.; Cohen, R. J.

1995-01-01

INTRODUCTION: Beat-to-beat adaptation of ventricular repolarization duration to cardiac cycle length and autonomic activity has not been previously characterized in the spontaneously beating human heart. METHODS AND RESULTS: The ECG of 14 healthy subjects was recorded from the supine and upright positions. Autonomic blockade was accomplished by atropine and propranolol. RR and RT intervals were measured by a computer algorithm, and the impulse response (h) from RR to RT computed. In the supine position the maximal adjustment of the RT interval occurred in the first beat following a change in cycle length (hpeak = 17.8 +/- 1.6 msec/sec), but continued to be detectable for 3.8 seconds (2.9-4.7 sec). Propranolol attenuated the peak impulse response to 15.8 +/- 4.0 msec/sec (P = NS). In the standing position the peak impulse response was increased to 25.2 +/- 5.0 msec/sec (P = 0.004 vs supine), and the impulse response duration (hdur) shortened to 1.4 seconds (1.3-1.6). This was reversed by beta blockade (hpeak = 10.7 +/- 3.6 [P = 0.005 vs standing]; hdur = 5.5 sec [4.8-6.1]). Parasympathetic and combined autonomic blockade resulted in too little residual heart rate variability to estimate the impulse response accurately. The slope of the regression of delta RT and delta RR in the supine position was 0.0177 +/- 0.0016, which was closely correlated with the peak impulse response (r = 0.91). CONCLUSIONS: System identification techniques can assist in characterizing the cycle dependence of ventricular repolarization and may provide new insights into conditions associated with abnormal repolarization.

Circulating anti-double-stranded DNA antibody-secreting cells in patients with systemic lupus erythematosus: a novel biomarker for disease activity.

PubMed

Hanaoka, H; Okazaki, Y; Satoh, T; Kaneko, Y; Yasuoka, H; Seta, N; Kuwana, M

2012-10-01

Antibodies against double-stranded DNA (dsDNA) are widely used to diagnose systemic lupus erythematosus (SLE) and evaluate its activity in patients. This study was undertaken to examine the clinical utility of circulating anti-dsDNA antibody-secreting cells for evaluating SLE patients. Anti-dsDNA antibody-secreting cells quantified using an enzyme-linked immunospot assay were detected in the spleen, bone marrow and peripheral blood from MRL/lpr but not in control BALB/c mice. Circulating anti-dsDNA antibody-secreting cells were detected in 29 (22%) of 130 patients with SLE, but in none of 49 with non-SLE connective-tissue disease or 18 healthy controls. The presence of circulating anti-dsDNA antibody-secreting cells was associated with persistent proteinuria, high SLE disease activity index and systemic lupus activity measures, and a high serum anti-dsDNA antibody titre measured with an enzyme-linked immunosorbent assay. The positive predictive value for active disease was 48% for circulating anti-dsDNA antibody-secreting cells versus 17% for serum anti-dsDNA antibodies. A prospective cohort of patients with circulating anti-dsDNA antibodies and inactive SLE showed that the cumulative disease flare-free rate was significantly lower in patients with than without circulating anti-dsDNA antibody-secreting cells (p < 0.001). Circulating anti-dsDNA antibody-secreting cells are a useful biomarker for assessing disease activity in SLE patients.

Functional Description for the Department of the Army Movements Management System. Redesign Phase 1. (DAMMS-R1). Volume 3.

DTIC Science & Technology

1987-12-31

Maintan -.ountry-TbI e CARDINALiii 134; HL~.,N-L.". 1); bL t i 11L h I...MEMO: FCityCd-Memo SOURCE IS: ’TMAS DAlA MODEL’ ATTRIBUTE IS: MEDIA ’DISK’ SEC-CLASS ’UNCLASSIFIED’ volatility ’DYNAMIC’I UPDATE- METHOD ’INTERACTIVE...8217 ATTRIBUTE IS: MEDIA ’DISK’ SEC-CLASS ’UNCLASSIFIED’ volatility ’STATTO’ UPDATE- METHOD ’INTERACTIVE’ UPDATE-FREQUENCY ’AS PER CHANGE’ Tt.LLE-FORM ’COMMAND’

Changes in particle transport as a result of resonant magnetic perturbations in DIII-D (vol 19, 056503, 2012)

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

Mordijck, S.; Doyle, E. J.; McKee, G. R.

2012-01-01

Publisher s Note: Changes in particle transport as a result of resonant magnetic perturbations in DIII-D [Phys. Plasmas 19, 056503 (2012)]a) In the Invited Papers from the 53rd Annual Meeting of the APS Division of Plasma Physics of the May 2012 issue of the journal, this article was originally published online and in print in the incorrect section; it was published within Ionospheric, Solar-System and Astrophysical Plasmas (Sec. 65) instead of Magnetically Confined Plasmas, Heating, Confinement (Sec. 61). AIP apologizes for this error. a)

Two-Party secret key distribution via a modified quantum secret sharing protocol

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

Grice, Warren P.; Evans, Philip G.; Lawrie, Benjamin

We present and demonstrate a method of distributing secret information based on N-party single-qubit Quantum Secret Sharing (QSS) in a modied plug-and-play two-party Quantum Key Distribution (QKD) system with N 2 intermediate nodes and compare it to both standard QSS and QKD. Our setup is based on the Clavis2 QKD system built by ID Quantique but is generalizable to any implementation. We show that any two out of N parties can build a secret key based on partial information from each other and with collaboration from the remaining N 2 parties. This method signicantly reduces the number of resources (singlemore » photon detectors, lasers and dark ber connections) needed to implement QKD on the grid.« less

Two-Party secret key distribution via a modified quantum secret sharing protocol

DOE PAGES

Grice, Warren P.; Evans, Philip G.; Lawrie, Benjamin; ...

2015-01-01

We present and demonstrate a method of distributing secret information based on N-party single-qubit Quantum Secret Sharing (QSS) in a modied plug-and-play two-party Quantum Key Distribution (QKD) system with N 2 intermediate nodes and compare it to both standard QSS and QKD. Our setup is based on the Clavis2 QKD system built by ID Quantique but is generalizable to any implementation. We show that any two out of N parties can build a secret key based on partial information from each other and with collaboration from the remaining N 2 parties. This method signicantly reduces the number of resources (singlemore » photon detectors, lasers and dark ber connections) needed to implement QKD on the grid.« less

The Role of Pathogen-Secreted Proteins in Fungal Vascular Wilt Diseases

PubMed Central

de Sain, Mara; Rep, Martijn

2015-01-01

A limited number of fungi can cause wilting disease in plants through colonization of the vascular system, the most well-known being Verticillium dahliae and Fusarium oxysporum. Like all pathogenic microorganisms, vascular wilt fungi secrete proteins during host colonization. Whole-genome sequencing and proteomics screens have identified many of these proteins, including small, usually cysteine-rich proteins, necrosis-inducing proteins and enzymes. Gene deletion experiments have provided evidence that some of these proteins are required for pathogenicity, while the role of other secreted proteins remains enigmatic. On the other hand, the plant immune system can recognize some secreted proteins or their actions, resulting in disease resistance. We give an overview of proteins currently known to be secreted by vascular wilt fungi and discuss their role in pathogenicity and plant immunity. PMID:26473835

Lifetime of the Excited State In Vivo

PubMed Central

Mar, T.; Govindjee; Singhal, G. S.; Merkelo, H.

1972-01-01

Using a mode-locked laser (λ, 632.8 nm), fluorescence decay of chlorophyll (Chl) a in the green alga Chlorella pyrenoidosa, the red alga Porphyridium cruentum, and the blue-green alga Anacystis nidulans was measured by the phase-shift method under conditions when photosynthesis was not operative (3-(3,4-dichlorophenyl)-1,1-dimethylurea [DCMU] poisoning, or cooling to 77°K). In the presence of 10-5 M DCMU, the lifetime of Chl a fluorescence (τ) at room temperature is about 1.7 nsec in Chlorella, 1.0 nsec in Porphyridium, and 0.7 nsec in Anacystis. At 77°K, τ is 1.4 nsec (for fluorescence at about 685 nm, F-685) and 2.3 nsec (for F-730) in Chlorella, 0.9 nsec (F-685) and 1.2 nsec (F-730) in Porphyridium, and 0.8 nsec (F-685 and F-730) in Anacystis. From the above measurement, and the assumption that τ0 (the intrinsic fluorescence lifetime) for Chl a in all three algae is 15.2 nsec, we have calculated the rate constants of radiationless transition (that includes energy transfer to weakly fluorescent system I) processes competing with fluorescence at room temperature to be about 5 × 108 sec-1 in Chlorella, 9 × 108 sec-1 in Porphyridium, and 13 × 108 sec-1 in Anacystis. At 77°K, this rate constant for Chl a that fluoresces at 685 nm remains, in the first approximation, the same as at room temperature. From the τ data, the rate constant for the trapping of excitation energy is calculated to be about 1.2 × 109 sec-1 for Chlorella, 2 × 109 sec-1 for Porphyridium, and 2 × 109 sec-1 for Anacystis. The efficiency of trapping is calculated to be about 66% (Chlorella), 68% (Porphyridium), and 60% (Anacystis). (It is recognized that variations in the above values are to be expected if algae grown under different conditions are used for experimentation.) The maximum quantum yield of Chl a fluorescence for system II (λ, 632.8 nm), calculated from τ measurements, is about 10% in Chlorella, 6-7% in Porhyridium, and 5% in Anacystis under conditions when photosynthesis is not operative; the values at 77°K appear to be very close to those with DCMU added at room temperature. ø for F-730 at 77°K, however, is somewhat higher than for F-685. The predicted quantum yields of fluorescence for Chl a in intact cells (both systems I and II) at low intensities of 632.8 nm light are about 2-3, 1-2, and 1% for Chlorella, Porphyridium, and Anacystis, respectively. PMID:4624832

78 FR 51742 - Notice of Proposed Withdrawal and Opportunity for Public Meeting; California

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Research Directions in Database Security, II

DTIC Science & Technology

1990-11-01

WILLIAMS Burke Ct 286 WOLCOTT 9th Rd 795 WOOD 25th St 520 YANCEY Motley St 398 ZUZACK Arden Rd LDV> : The style of the prototype is such that the...WOLCOTT Fin Clk YANCEY Dept Mgr ZUZACK Proc Anal LDV> ;RR3: S (sortrel (project (njoin (project employee-base ’(Department Employee-Name Employee-Num...Proc Anal PROC MAHONEY Secy PROC YANCEY Dept Mgr PROC ZUZACK Proc Anal SEC BRIMER DMSO SEC FALBO Secy SEC HILL Dept Mgr SEC MITCHELL Ast DBSO SEC THOMAS

C-tactile afferent stimulating touch carries a positive affective value.

PubMed

Pawling, Ralph; Cannon, Peter R; McGlone, Francis P; Walker, Susannah C

2017-01-01

The rewarding sensation of touch in affiliative interactions is hypothesized to be underpinned by a specialized system of nerve fibers called C-Tactile afferents (CTs), which respond optimally to slowly moving, gentle touch, typical of a caress. However, empirical evidence to support the theory that CTs encode socially relevant, rewarding tactile information in humans is currently limited. While in healthy participants, touch applied at CT optimal velocities (1-10cm/sec) is reliably rated as subjectively pleasant, neuronopathy patients lacking large myelinated afferents, but with intact C-fibres, report that the conscious sensation elicited by stimulation of CTs is rather vague. Given this weak perceptual impact the value of self-report measures for assessing the specific affective value of CT activating touch appears limited. Therefore, we combined subjective ratings of touch pleasantness with implicit measures of affective state (facial electromyography) and autonomic arousal (heart rate) to determine whether CT activation carries a positive affective value. We recorded the activity of two key emotion-relevant facial muscle sites (zygomaticus major-smile muscle, positive affect & corrugator supercilii-frown muscle, negative affect) while participants evaluated the pleasantness of experimenter administered stroking touch, delivered using a soft brush, at two velocities (CT optimal 3cm/sec & CT non-optimal 30cm/sec), on two skin sites (CT innervated forearm & non-CT innervated palm). On both sites, 3cm/sec stroking touch was rated as more pleasant and produced greater heart rate deceleration than 30cm/sec stimulation. However, neither self-report ratings nor heart rate responses discriminated stimulation on the CT innervated arm from stroking of the non-CT innervated palm. In contrast, significantly greater activation of the zygomaticus major (smiling muscle) was seen specifically to CT optimal, 3cm/sec, stroking on the forearm in comparison to all other stimuli. These results offer the first empirical evidence in humans that tactile stimulation that optimally activates CTs carries a positive affective valence that can be measured implicitly.

Final Technical Report for Year 5 Early Career Research Project "Viscosity and equation of state of hot and dense QCD matter"

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

Molnar, Denes

2016-05-25

The Section below summarizes research activities and achievements during the fifth (last) year of the PI’s Early Career Research Project (ECRP). Unlike the first four years of the project, the last year was not funded under the American Recovery and Reinvestment Act (ARRA). The ECRP advanced two main areas: i) radiative 3 ↔ 2 radiative transport, via development of a new computer code MPC/Grid that solves the Boltzmann transport equation in full 6+1D (3X+3V+time); and ii) application of relativistic hydrodynamics, via development of a self-consistent framework to convert viscous fluids to particles. In Year 5 we finalized thermalization studies withmore » radiative gg ↔ ggg transport (Sec. 1.1.1) and used nonlinear covariant transport to assess the accuracy of fluid-to-particle conversion models (Sec. 1.1.2), calculated observables with self-consistent fluid-to-particle conversion from realistic viscous hydrodynamic evolution (Secs. 1.2.1 and 1.2.2), extended the covariant energy loss formulation to heavy quarks (Sec. 1.4.1) and studied energy loss in small systems (Sec. 1.4.2), and also investigated how much of the elliptic flow could have non-hydrodynamic origin (Sec 1.3). Years 1-4 of the ECRP were ARRA-funded and, therefore, they have their own report document ’Final Technical Report for Years 1-4 of the Early Career Research Project “Viscosity and equation of state of hot and dense QCD matter”’ (same award number DE-SC0004035). The PI’s group was also part of the DOE JET Topical Collaboration, a multi-institution project that overlapped in time significantly with the ECRP. Purdue achievements as part of the JET Top- ical Collaboration are in a separate report “Final Technical Report summarizing Purdue research activities as part of the DOE JET Topical Collaboration” (award DE-SC0004077).« less

The design and evaluation of grazing incidence relay optics

NASA Technical Reports Server (NTRS)

Davis, John M.; Chase, R. C.; Silk, J. K.; Krieger, A. S.

1989-01-01

X-ray astronomy, both solar and celestial, has many needs for high spatial resolution observations which have to be performed with electronic detectors. If the resolution is not to be detector limited, plate scales in excess of 25 microns arc/sec, corresponding to focal lengths greater than 5 m, are required. In situations where the physical size is restricted, the problem can be solved by the use of grazing incidence relay optics. A system was developed which employs externally polished hyperboloid-hyperboloid surfaces to be used in conjunction with a Wolter-Schwarzschild primary. The secondary is located in front of the primary focus and provides a magnification of 4, while the system has a plate scale of 28 microns arc/sec and a length of 1.9 m. The design, tolerance specification, fabrication and performance at visible and X-ray wavelengths of this optical system are described.

Stromal Cells Positively and Negatively Modulate the Growth of Cancer Cells: Stimulation via the PGE2-TNFα-IL-6 Pathway and Inhibition via Secreted GAPDH-E-Cadherin Interaction

PubMed Central

Kawada, Manabu; Inoue, Hiroyuki; Ohba, Shun-ichi; Yoshida, Junjiro; Masuda, Tohru; Yamasaki, Manabu; Usami, Ihomi; Sakamoto, Shuichi; Abe, Hikaru; Watanabe, Takumi; Yamori, Takao; Shibasaki, Masakatsu; Nomoto, Akio

2015-01-01

Fibroblast-like stromal cells modulate cancer cells through secreted factors and adhesion, but those factors are not fully understood. Here, we have identified critical stromal factors that modulate cancer growth positively and negatively. Using a cell co-culture system, we found that gastric stromal cells secreted IL-6 as a growth and survival factor for gastric cancer cells. Moreover, gastric cancer cells secreted PGE2 and TNFα that stimulated IL-6 secretion by the stromal cells. Furthermore, we found that stromal cells secreted glyceraldehyde 3-phosphate dehydrogenase (GAPDH). Extracellular GAPDH, or its N-terminal domain, inhibited gastric cancer cell growth, a finding confirmed in other cell systems. GAPDH bound to E-cadherin and downregulated the mTOR-p70S6 kinase pathway. These results demonstrate that stromal cells could regulate cancer cell growth through the balance of these secreted factors. We propose that negative regulation of cancer growth using GAPDH could be a new anti-cancer strategy. PMID:25785838

LegC3, an Effector Protein from Legionella pneumophila, Inhibits Homotypic Yeast Vacuole Fusion In Vivo and In Vitro

PubMed Central

Bennett, Terry L.; Kraft, Shannon M.; Reaves, Barbara J.; Mima, Joji; O’Brien, Kevin M.; Starai, Vincent J.

2013-01-01

During infection, the intracellular pathogenic bacterium Legionella pneumophila causes an extensive remodeling of host membrane trafficking pathways, both in the construction of a replication-competent vacuole comprised of ER-derived vesicles and plasma membrane components, and in the inhibition of normal phagosome:endosome/lysosome fusion pathways. Here, we identify the LegC3 secreted effector protein from L. pneumophila as able to inhibit a SNARE- and Rab GTPase-dependent membrane fusion pathway in vitro, the homotypic fusion of yeast vacuoles (lysosomes). This vacuole fusion inhibition appeared to be specific, as similar secreted coiled-coiled domain containing proteins from L. pneumophila, LegC7/YlfA and LegC2/YlfB, did not inhibit vacuole fusion. The LegC3-mediated fusion inhibition was reversible by a yeast cytosolic extract, as well as by a purified soluble SNARE, Vam7p. LegC3 blocked the formation of trans-SNARE complexes during vacuole fusion, although we did not detect a direct interaction of LegC3 with the vacuolar SNARE protein complexes required for fusion. Additionally, LegC3 was incapable of inhibiting a defined synthetic model of vacuolar SNARE-driven membrane fusion, further suggesting that LegC3 does not directly inhibit the activity of vacuolar SNAREs, HOPS complex, or Sec17p/18p during membrane fusion. LegC3 is likely utilized by Legionella to modulate eukaryotic membrane fusion events during pathogenesis. PMID:23437241

Proteomic analysis of enterotoxigenic Escherichia coli (ETEC) in neutral and alkaline conditions.

PubMed

Gonzales-Siles, Lucia; Karlsson, Roger; Kenny, Diarmuid; Karlsson, Anders; Sjöling, Åsa

2017-01-07

Enterotoxigenic Escherichia coli (ETEC) is a major cause of diarrhea in children and travelers to endemic areas. Secretion of the heat labile AB 5 toxin (LT) is induced by alkaline conditions. In this study, we determined the surface proteome of ETEC exposed to alkaline conditions (pH 9) as compared to neutral conditions (pH 7) using a LPI Hexalane FlowCell combined with quantitative proteomics. Relative quantitation with isobaric labeling (TMT) was used to compare peptide abundance and their corresponding proteins in multiple samples at MS/MS level. For protein identification and quantification samples were analyzed using either a 1D-LCMS or a 2D-LCMS approach. Strong up-regulation of the ATP synthase operon encoding F1Fo ATP synthase and down-regulation of proton pumping proteins NuoF, NuoG, Ndh and WrbA were detected among proteins involved in regulating the proton and electron transport under alkaline conditions. Reduced expression of proteins involved in osmotic stress was found at alkaline conditions while the Sec-dependent transport over the inner membrane and outer membrane protein proteins such as OmpA and the β-Barrel Assembly Machinery (BAM) complex were up-regulated. ETEC exposed to alkaline environments express a specific proteome profile characterized by up-regulation of membrane proteins and secretion of LT toxin. Alkaline microenvironments have been reported close to the intestinal epithelium and the alkaline proteome may hence represent a better view of ETEC during infection.

Cloning of the Pichia anomala SEC61 gene and its expression in a Saccharomyces cerevisiae sec61 mutant.

PubMed

Ruíz, Teresa; De la Rosa, José M; Domínguez, Angel; Rodríguez, Luis

2003-05-01

In several organisms, including Saccharomyces cerevisiae and other yeast species, the product encoded by the SEC61 gene is considered to be the core element of the translocation apparatus within the endoplasmic reticulum membrane through which translocation of secretory and membrane proteins occurs. In this study, we have cloned and characterized the homolog of the SEC61 gene from the yeast Pichia anomala. The cloned gene includes an ORF, interrupted after the first ten nucleotides by an intron of 131 bp, encoding a 479-amino acid putative polypeptide exhibiting homology to the products encoded by different eukaryotic SEC61 genes, particularly to those from other yeast species. We show that the P. anomala SEC61 gene is correctly processed (intron splicing) when expressed in S. cerevisiae and that it is able to complement the thermosensitive phenotype associated with a mutation in the S. cerevisiae SEC61 gene.

Sec63p and Kar2p are required for the translocation of SRP-dependent precursors into the yeast endoplasmic reticulum in vivo

PubMed Central

Young, Barry P.; Craven, Rachel A.; Reid, Peter J.; Willer, Martin; Stirling, Colin J.

2001-01-01

The translocation of secretory polypeptides into the endoplasmic reticulum (ER) occurs at the translocon, a pore-forming structure that orchestrates the transport and maturation of polypeptides at the ER membrane. In yeast, targeting of secretory precursors to the translocon can occur by two distinct pathways that are distinguished by their dependence upon the signal recognition particle (SRP). The SRP-dependent pathway requires SRP and its membrane-bound receptor, whereas the SRP-independent pathway requires a separate receptor complex consisting of Sec62p, Sec63p, Sec71p, Sec72p plus lumenal Kar2p/BiP. Here we demonstrate that Sec63p and Kar2p are also required for the SRP-dependent targeting pathway in vivo. Furthermore, we demonstrate multiple roles for Sec63p, at least one of which is exclusive to the SRP-independent pathway. PMID:11226176

ssaD1, a suppressor of secA51(Ts) that renders growth of Escherichia coli cold sensitive, is an early amber mutation in the transcription factor gene nusB.

PubMed Central

Rajapandi, T.; Oliver, D.

1994-01-01

Complementation analysis of the ssaD1 mutation, isolated as a suppressor of the secA51(Ts) mutation that renders growth of Escherichia coli cold sensitive, was used to show that ssaD corresponds to nusB, a gene known to be important in transcription antitermination. DNA sequence analysis of the ssaD1 allele showed that it creates an amber mutation in the 15th codon of nusB. Analysis of the effect of different levels of NusB protein on secA transcription and translation suggested that NusB plays little or no role in the control of secA expression. Accordingly, mechanisms by which nusB inactivation can lead to suppression of secA51(Ts) and secY24(Ts) mutations without affecting secA expression need to be considered. PMID:8021230

Structure of Salmonella FlhE, conserved member of a flagellar Type III secretion operon

DOE PAGES

Lee, Jaemin; Monzingo, Arthur F.; Keatinge-Clay, Adrian T.; ...

2014-12-26

In this paper, the bacterial flagellum is assembled by a multicomponent transport apparatus categorized as a type III secretion system. The secretion of proteins that assemble into the flagellum is driven by the proton motive force. The periplasmic protein FlhE is a member of the flhBAE operon in the majority of bacteria where FlhE is found. FlhA and FlhB are established components of the flagellar type III secretion system. The absence of FlhE results in a proton leak through the flagellar system, inappropriate secretion patterns, and cell death, indicating that FlhE regulates an important aspect of proper flagellar biosynthesis. Wemore » isolated FlhE from the periplasm of Salmonella and solved its structure to 1.5 Å resolution. The structure reveals a β-sandwich fold, with no close structural homologs. Finally, possible roles of FlhE, including that of a chaperone, are discussed.« less

Structure of Salmonella FlhE, conserved member of a flagellar Type III secretion operon

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

Lee, Jaemin; Monzingo, Arthur F.; Keatinge-Clay, Adrian T.

In this paper, the bacterial flagellum is assembled by a multicomponent transport apparatus categorized as a type III secretion system. The secretion of proteins that assemble into the flagellum is driven by the proton motive force. The periplasmic protein FlhE is a member of the flhBAE operon in the majority of bacteria where FlhE is found. FlhA and FlhB are established components of the flagellar type III secretion system. The absence of FlhE results in a proton leak through the flagellar system, inappropriate secretion patterns, and cell death, indicating that FlhE regulates an important aspect of proper flagellar biosynthesis. Wemore » isolated FlhE from the periplasm of Salmonella and solved its structure to 1.5 Å resolution. The structure reveals a β-sandwich fold, with no close structural homologs. Finally, possible roles of FlhE, including that of a chaperone, are discussed.« less

76 FR 10086 - Petition for Waiver of Compliance

Federal Register 2010, 2011, 2012, 2013, 2014

2011-02-23

... Railroad Freight Car Safety Standards, i.e. Sec. Sec. 215.303 and 215.305, which require stenciling of restricted cars; as well as that of the Reflectorization of Rail Freight Rolling Stock, i.e. Sec. Sec. 224.3... requirements of its safety standards. The individual petition is described below, including the party seeking...

75 FR 44781 - Joint CFTC-SEC Advisory Committee on Emerging Regulatory Issues

Federal Register 2010, 2011, 2012, 2013, 2014

2010-07-29

...; File No. 265-26] Joint CFTC-SEC Advisory Committee on Emerging Regulatory Issues AGENCY: Commodity... Meeting of Joint CFTC-SEC Advisory Committee on Emerging Regulatory Issues. SUMMARY: The Joint CFTC-SEC Advisory Committee on Emerging Regulatory Issues will hold a public meeting on August 11, 2010, from 9 a.m...

75 FR 66362 - Joint CFTC-SEC Advisory Committee on Emerging Regulatory Issues

Federal Register 2010, 2011, 2012, 2013, 2014

2010-10-28

...; File No. 265-26] Joint CFTC-SEC Advisory Committee on Emerging Regulatory Issues AGENCY: Commodity... meeting of Joint CFTC-SEC Advisory Committee on Emerging Regulatory Issues. SUMMARY: The Joint CFTC-SEC Advisory Committee on Emerging Regulatory Issues will hold a public meeting on November 5, 2010, from 9 a.m...

Engraftment and reconstitution of hematopoiesis is dependent on VEGFR2 mediated regeneration of sinusoidal endothelial cells

PubMed Central

Hooper, Andrea T.; Butler, Jason M.; Nolan, Daniel J; Kranz, Andrea; Iida, Kaoruko; Kobayashi, Mariko; Kopp, Hans-Georg; Shido, Koji; Petit, Isabelle; Yanger, Kilangsungla; James, Daylon; Witte, Larry; Zhu, Zhenping; Wu, Yan; Pytowski, Bronislaw; Rosenwaks, Zev; Mittal, Vivek; Sato, Thomas N.; Rafii, Shahin

2011-01-01

SUMMARY The phenotypic attributes and molecular determinants for the regeneration of bone marrow (BM) sinusoidal endothelial cells (SECs) and their contribution to hematopoiesis are unknown. We show that after myelosuppression VEGFR2 activation promotes reassembly of regressed SECs, reconstituting hematopoietic stem and progenitor cells (HSPCs). VEGFR2 and VEGFR3 expression are restricted to BM vasculature, demarcating a continuous network of VEGFR2+VEGFR3+Sca1− SECs and VEGFR2+VEGFR3−Sca1+ arterioles. While chemotherapy (5FU) and sublethal irradiation (650 rad) induce minor SEC regression, lethal irradiation (950 rad) induces severe regression of SECs requiring BM transplantation (BMT) for regeneration. Conditional deletion of VEGFR2 in adult mice blocks regeneration of SECs in sublethally irradiated animals, preventing hematopoietic reconstitution. Inhibition of VEGFR2 signaling in lethally irradiated wild type mice rescued with BMT severely impairs SEC reconstruction, preventing engraftment and reconstitution of HSPCs. Therefore, activation of VEGFR2 is critical for regeneration of VEGFR3+Sca1− SECs that are essential for engraftment and restoration of HSPCs and hematopoiesis. PMID:19265665

Preferential Targeting of a Signal Recognition Particle-dependent Precursor to the Ssh1p Translocon in Yeast♦

PubMed Central

Spiller, Michael P.; Stirling, Colin J.

2011-01-01

Protein translocation across the endoplasmic reticulum membrane occurs via a “translocon” channel formed by the Sec61p complex. In yeast, two channels exist: the canonical Sec61p channel and a homolog called Ssh1p. Here, we used trapped translocation intermediates to demonstrate that a specific signal recognition particle-dependent substrate, Sec71p, is targeted exclusively to Ssh1p. Strikingly, we found that, in the absence of Ssh1p, precursor could be successfully redirected to canonical Sec61p, demonstrating that the normal targeting reaction must involve preferential sorting to Ssh1p. Our data therefore demonstrate that Ssh1p is the primary translocon for Sec71p and reveal a novel sorting mechanism at the level of the endoplasmic reticulum membrane enabling precursors to be directed to distinct translocons. Interestingly, the Ssh1p-dependent translocation of Sec71p was found to be dependent upon Sec63p, demonstrating a previously unappreciated functional interaction between Sec63p and the Ssh1p translocon. PMID:21454595

Inhibition of a type III secretion system by the deletion of a short loop in one of its membrane proteins

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

Meshcheryakov, Vladimir A.; Kitao, Akio; Core Research for Evolutionary Science and Technology, Tokyo 113-0032

2013-05-01

Crystal structures of the cytoplasmic domain of FlhB from S. typhimurium and A. aeolicus were solved at 2.45 and 2.55 Å resolution, respectively. The deletion of a short loop in the cytoplasmic domain of Salmonella FlhB completely abolishes secretion by the type III secretion system. A molecular-dynamics simulation shows that the deletion of the loop affects the flexibility of a linker between the transmembrane and cytoplasmic domains of FlhB. The membrane protein FlhB is a highly conserved component of the flagellar secretion system. It is composed of an N-terminal transmembrane domain and a C-terminal cytoplasmic domain (FlhB{sub C}). Here, themore » crystal structures of FlhB{sub C} from Salmonella typhimurium and Aquifex aeolicus are described at 2.45 and 2.55 Å resolution, respectively. These flagellar FlhB{sub C} structures are similar to those of paralogues from the needle type III secretion system, with the major difference being in a linker that connects the transmembrane and cytoplasmic domains of FlhB. It was found that deletion of a short flexible loop in a globular part of Salmonella FlhB{sub C} leads to complete inhibition of secretion by the flagellar secretion system. Molecular-dynamics calculations demonstrate that the linker region is the most flexible part of FlhB{sub C} and that the deletion of the loop reduces this flexibility. These results are in good agreement with previous studies showing the importance of the linker in the function of FlhB and provide new insight into the relationship between the different parts of the FlhB{sub C} molecule.« less

Testing the scalar expectancy theory (SET) and the learning-to-time model (LeT) in a double bisection task.

PubMed

Machado, Armando; Pata, Paulo

2005-02-01

Two theories of timing, scalar expectancy theory (SET) and learning-to-time (LeT), make substantially different assumptions about what animals learn in temporal tasks. In a test of these assumptions, pigeons learned two temporal discriminations. On Type 1 trials, they learned to choose a red key after a 1-sec signal and a green key after a 4-sec signal; on Type 2 trials, they learned to choose a blue key after a 4-sec signal and a yellow key after either an 8-sec signal (Group 8) or a 16-sec signal (Group 16). Then, the birds were exposed to signals 1 sec, 4 sec, and 16 sec in length and given a choice between novel key combinations (red or green vs. blue or yellow). The choice between the green key and the blue key was of particular significance because both keys were associated with the same 4-sec signal. Whereas SET predicted no effect of the test signal duration on choice, LeT predicted that preference for green would increase monotonically with the length of the signal but would do so faster for Group 8 than for Group 16. The results were consistent with LeT, but not with SET.

Native structure of a type IV secretion system core complex essential for Legionella pathogenesis.

PubMed

Kubori, Tomoko; Koike, Masafumi; Bui, Xuan Thanh; Higaki, Saori; Aizawa, Shin-Ichi; Nagai, Hiroki

2014-08-12

Bacterial type IV secretion systems are evolutionarily related to conjugation systems and play a pivotal role in infection by delivering numerous virulence factors into host cells. Using transmission electron microscopy, we report the native molecular structure of the core complex of the Dot/Icm type IV secretion system encoded by Legionella pneumophila, an intracellular human pathogen. The biochemically isolated core complex, composed of at least five proteins--DotC, DotD, DotF, DotG, and DotH--has a ring-shaped structure. Intriguingly, morphologically distinct premature complexes are formed in the absence of DotG or DotF. Our data suggest that DotG forms a central channel spanning inner and outer membranes. DotF, a component dispensable for type IV secretion, plays a role in efficient embedment of DotG into the functional core complex. These results highlight a common scheme for the biogenesis of transport machinery.

Regulation of pulmonary surfactant secretion in the developing lizard, Pogona vitticeps.

PubMed

Sullivan, Lucy C; Orgeig, Sandra; Daniels, Christopher B

2002-11-01

Pulmonary surfactant is a mixture of lipids and proteins that is secreted by alveolar type II cells in the lungs of all air-breathing vertebrates. Pulmonary surfactant functions to reduce the surface tension in the lungs and, therefore, reduce the work of breathing. In mammals, the embryonic maturation of the surfactant system is controlled by a host of factors, including glucocorticoids, thyroid hormones and autonomic neurotransmitters. We have used a co-culture system of embryonic type II cells and lung fibroblasts to investigate the ability of dexamethasone, tri-iodothyronine (T(3)), adrenaline and carbamylcholine (carbachol) to stimulate the cellular secretion of phosphatidylcholine in the bearded dragon (Pogona vitticeps) at day 55 (approx. 92%) of incubation and following hatching. Adrenaline stimulated surfactant secretion both before and after hatching, whereas carbachol stimulated secretion only at day 55. Glucocorticoids and triiodothyronine together stimulated secretion at day 55 but did not after hatching. Therefore, adrenaline, carbachol, dexamethasone and T(3), are all involved in the development of the surfactant system in the bearded dragon. However, the efficacy of the hormones is attenuated during the developmental process. These differences probably relate to the changes in the cellular environment during development and the specific biology of the bearded dragon.

A FRET-Based DNA Biosensor Tracks OmpR-Dependent Acidification of Salmonella during Macrophage Infection

PubMed Central

Chakraborty, Smarajit; Mizusaki, Hideaki; Kenney, Linda J.

2015-01-01

In bacteria, one paradigm for signal transduction is the two-component regulatory system, consisting of a sensor kinase (usually a membrane protein) and a response regulator (usually a DNA binding protein). The EnvZ/OmpR two-component system responds to osmotic stress and regulates expression of outer membrane proteins. In Salmonella, EnvZ/OmpR also controls expression of another two-component system SsrA/B, which is located on Salmonella Pathogenicity Island (SPI) 2. SPI-2 encodes a type III secretion system, which functions as a nanomachine to inject bacterial effector proteins into eukaryotic cells. During the intracellular phase of infection, Salmonella switches from assembling type III secretion system structural components to secreting effectors into the macrophage cytoplasm, enabling Salmonella to replicate in the phagocytic vacuole. Major questions remain regarding how bacteria survive the acidified vacuole and how acidification affects bacterial secretion. We previously reported that EnvZ sensed cytoplasmic signals rather than extracellular ones, as intracellular osmolytes altered the dynamics of a 17-amino-acid region flanking the phosphorylated histidine. We reasoned that the Salmonella cytoplasm might acidify in the macrophage vacuole to activate OmpR-dependent transcription of SPI-2 genes. To address these questions, we employed a DNA-based FRET biosensor (“I-switch”) to measure bacterial cytoplasmic pH and immunofluorescence to monitor effector secretion during infection. Surprisingly, we observed a rapid drop in bacterial cytoplasmic pH upon phagocytosis that was not predicted by current models. Cytoplasmic acidification was completely dependent on the OmpR response regulator, but did not require known OmpR-regulated genes such as ompC, ompF, or ssaC (SPI-2). Microarray analysis highlighted the cadC/BA operon, and additional experiments confirmed that it was repressed by OmpR. Acidification was blocked in the ompR null background in a Cad-dependent manner. Acid-dependent activation of OmpR stimulated type III secretion; blocking acidification resulted in a neutralized cytoplasm that was defective for SPI-2 secretion. Based upon these findings, we propose that Salmonella infection involves an acid-dependent secretion process in which the translocon SseB moves away from the bacterial cell surface as it associates with the vacuolar membrane, driving the secretion of SPI-2 effectors such as SseJ. New steps in the SPI-2 secretion process are proposed. PMID:25875623

A 72-hour study to test the efficacy and safety of the "Mucus Slurper" in mechanically ventilated sheep.

PubMed

Li Bassi, Gianluigi; Curto, Francesco; Zanella, Alberto; Stylianou, Mario; Kolobow, Theodor

2007-03-01

In patients intubated with endotracheal tubes (ETTs), suctioning is routinely performed to remove mucus from within the ETT and trachea. The Mucus Slurper is a novel ETT with built-in suction ports arranged radially at its tip. We evaluated the safety and efficacy of the Mucus Slurper, compared with conventional tracheal suction, to prevent airway obstruction in sheep with the ETT and trachea oriented below horizontal. Prospective randomized animal study. Animal research facility at the National Institutes of Health. Twelve healthy sheep. Sheep were randomized to be intubated with either the Mucus Slurper (study group) or a Hi-Lo Tracheal Tube (Mallinckrodt, St. Louis, MO) (control group) and mechanically ventilated for 72 hrs. In the study group, automatic, timed tracheal aspiration lasted 0.3 secs, was repeated every 2 mins, and was synchronized with the early expiratory phase. In the control group, tracheal suction was performed every 6 hrs or as required. In the control group, tracheal secretions accumulated progressively within the ETT and the trachea. In the study group, all mucus that reached the tip of the Mucus Slurper was aspirated, keeping the lumen of the ETT, and proximal trachea, free from secretions. In the study group, expiratory water trap protein concentration, a crude index to measure mucus drainage through the ETT, was consistently less than the control group (p < .001). At autopsy, no macroscopic injury to the tracheal mucosa was found in either group. In the study group, the respiratory circuit was less colonized than in the control group. There were no statistically significant differences between the two groups in bacterial colonization of the lungs/bronchi. The Mucus Slurper, combined with orientation of the trachea below horizontal, prevents accumulation of secretions within the lumen of the ETT and trachea, without need for conventional tracheal suctioning.

Correlating levels of type III secretion and secreted proteins with fecal shedding of Escherichia coli O157:H7 in cattle

USDA-ARS?s Scientific Manuscript database

The locus of enterocyte effacement (LEE) encodes a type III secretion system (T3SS) for secreting factors that enable Escherichia coli O157:H7 to produce attaching and effacing lesions (A/E) on epithelial cells. The importance of LEE-encoded proteins in intestinal colonization of cattle is well-stud...

The effect of interplanetary trajectory options on a manned Mars aerobrake configuration

NASA Technical Reports Server (NTRS)

Braun, Robert D.; Powell, Richard W.; Hartung, Lin C.

1990-01-01

Manned Mars missions originating in low Earth orbit (LEO) in the time frame 2010 to 2025 were analyzed to identify preferred mission opportunities and their associated vehicle and trajectory characteristics. Interplanetary and Mars atmospheric trajectory options were examined under the constraints of an initial manned exploration scenario. Two chemically propelled vehicle options were considered: (1) an all propulsive configuration, and (2) a configuration which employs aerobraking at Earth and Mars with low lift/drag (L/D) shapes. Both the interplanetary trajectory options as well as the Mars atmospheric passage are addressed to provide a coupled trajectory simulation. Direct and Venus swingby interplanetary transfers with a 60 day Mars stopover are considered. The range and variation in both Earth and Mars entry velocity are also defined. Two promising mission strategies emerged from the study: (1) a 1.0 to 2.0 year Venus swingby mission, and (2) a 2.0 to 2.5 year direct mission. Through careful trajectory selection, 11 mission opportunities are identified in which the Mars entry velocity is between 6 and 10 km/sec and Earth entry velocity ranges from 11.5 to 12.5 km/sec. Simulation of the Earth return aerobraking maneuver is not performed. It is shown that a low L/D configuration is not feasible for Mars aerobraking without substantial improvements in the interplanetary navigation system. However, even with an advanced navigation system, entry corridor and aerothermal requirements restrict the number of potential mission opportunities. It is also shown that for a large blunt Mars aerobrake configuration, the effects of radiative heating can be significant at entry velocities as low as 6.2 km/sec and will grow to dominate the aerothermal environment at entry velocities above 8.5 km/sec. Despite the additional system complexity associated with an aerobraking vehicle, the use of aerobraking was shown to significantly lower the required initial LEO weight. In comparison with an all propulsive mission, savings between 19 and 59 percent were obtained depending upon launch date.

Selenocysteine Confers Resistance to Inactivation by Oxidation in Thioredoxin Reductase: Comparison of Selenium and Sulfur Enzymes†

PubMed Central

Snider, Gregg W.; Ruggles, Erik; Khan, Nadeem; Hondal, Robert J.

2013-01-01

Mammalian thioredoxin reductase (TR) is a selenocysteine (Sec)-containing homodimeric pyridine nucleotide oxidoreductase which catalyzes the reduction of oxidized thioredoxin. We have previously demonstrated the full-length mitochondrial mammalian TR (mTR3) enzyme to be resistant to inactivation from exposure to 50 mM H2O2. Because a Sec residue oxidizes more rapidly than a cysteine (Cys) residue, it has been previously thought that Sec-containing enzymes are “sensitive to oxidation” compared to Cys-orthologs. Here we show for the first time a direct comparison of the abilities of Sec-containing mTR3 and the Cys-ortholog from D. melanogaster (DmTR) to resist inactivation by oxidation from a variety of oxidants including H2O2, hydroxyl radical, peroxynitrite, hypochlorous acid, hypobromous acid, and hypothiocyanous acid. The results show that the Sec-containing TR is far superior to the Cys-ortholog TR in resisting inactivation by oxidation. To further test our hypothesis that the use of Sec confers strong resistance to inactivation by oxidation, we constructed a chimeric enzyme in which we replaced the active site Cys nucleophile of DmTR with a Sec residue using semisynthesis. The chimeric Sec-containing enzyme has similar ability to resist inactivation by oxidation as the wild type Sec-containing TR from mouse mitochondria. The use of Sec in the chimeric enzyme “rescued” the enzyme from oxidant-induced inactivation for all of the oxidants tested in this study, in direct contrast to previous understanding. We discuss two possibilities for this rescue effect from inactivation under identical conditions of oxidative stress: (i) Sec resists over-oxidation and inactivation, whereas a Cys residue can be permanently over-oxidized to the sulfinic acid form, and (ii) Sec protects the body of the enzyme from harmful oxidation by allowing the enzyme to metabolize (turnover) various oxidants much better than a Cys-containing TR. PMID:23865454

Shear-wave elastography of the breast: value of a quality measure and comparison with strain elastography.

PubMed

Barr, Richard G; Zhang, Zheng

2015-04-01

To determine whether addition of quality measure (QM) of shear-wave (SW) velocity (Vs) estimation can increase SW elastography sensitivity for breast cancer. With written informed consent, this institutional review board-approved, HIPAA-compliant study included 143 women (mean age, 48.5 years ± 8.7) scheduled for breast biopsy. Mean lesion size was 16.4 mm ± 11.8; 95 (66%) lesions were benign; 48 (34%), malignant. If more than one lesion was present, lesion with highest Breast Imaging Reporting and Data System (BI-RADS) category was chosen. If there were more than one with highest BI-RADS category, a lesion was randomly selected. Conventional ultrasonography (US), strain elastography, and SW elastography were performed with QM. QM assesses SW quality to provide accurate Vs. Lesions were evaluated for Vs and QM (high or low). Lesions with Vs of less than 4.5 m/sec were classified benign; lesions with Vs of 4.5 m/sec or greater, malignant. Results were correlated with pathologic findings. Vs data with or without incorporating QM were used to determine SW elastography diagnostic performance. Binomial proportions and exact 95% confidence intervals (CIs) were calculated. In 95 benign lesions, 13 (14%) had no SW elastography signal; 77 (81%), Vs of less than 4.5 m/sec; and five (5%), Vs of 4.5 m/sec or greater. In 48 malignant lesions, eight (17%) had no SW elastography signal; 20 (42%), Vs of less than 4.5 m/sec; and 20 (42%), V of 4.5 m/sec or greater. QM was low in 17 of 20 (85%) malignant lesions with Vs of less than 4.5 m/sec. Without QM, using Vs of 4.5 m/sec or greater as test positive, SW elastography had lesion-level sensitivity of 50% (95% CI: 34%, 66%); specificity, 94% (95% CI: 86%, 98%); positive predictive value (PPV), 80% (95% CI: 59%, 93%); and negative predictive value (NPV), 79% (95% CI: 70%, 87%). Using QM where additional lesions with both low Vs and low QM were treated as test positive, SW elastography had lesion-level sensitivity of 93% (95% CI: 80%, 98%); specificity, 89% (95% CI: 80%, 95%); PPV, 80% (95% CI: 66%, 91%); and NPV, 96% (95% CI: 89%, 99%). Addition of QM can improve SW elastography sensitivity, with no significant change in specificity. © RSNA, 2014 Online supplemental material is available for this article.