Sample records for cop operon required

  1. The copYAZ Operon Functions in Copper Efflux, Biofilm Formation, Genetic Transformation, and Stress Tolerance in Streptococcus mutans

    PubMed Central

    Singh, Kamna; Senadheera, Dilani B.; Lévesque, Céline M.

    2015-01-01

    ABSTRACT In bacteria, copper homeostasis is closely monitored to ensure proper cellular functions while avoiding cell damage. Most Gram-positive bacteria utilize the copYABZ operon for copper homeostasis, where copA and copB encode copper-transporting P-type ATPases, whereas copY and copZ regulate the expression of the cop operon. Streptococcus mutans is a biofilm-forming oral pathogen that harbors a putative copper-transporting copYAZ operon. Here, we characterized the role of copYAZ operon in the physiology of S. mutans and delineated the mechanisms of copper-induced toxicity in this bacterium. We observed that copper induced toxicity in S. mutans cells by generating oxidative stress and disrupting their membrane potential. Deletion of the copYAZ operon in S. mutans strain UA159 resulted in reduced cell viability under copper, acid, and oxidative stress relative to the viability of the wild type under these conditions. Furthermore, the ability of S. mutans to form biofilms and develop genetic competence was impaired under copper stress. Briefly, copper stress significantly reduced cell adherence and total biofilm biomass, concomitantly repressing the transcription of the gtfB, gtfC, gtfD, gbpB, and gbpC genes, whose products have roles in maintaining the structural and/or functional integrity of the S. mutans biofilm. Furthermore, supplementation with copper or loss of copYAZ resulted in significant reductions in transformability and in the transcription of competence-associated genes. Copper transport assays revealed that the ΔcopYAZ strain accrued significantly large amounts of intracellular copper compared with the amount of copper accumulation in the wild-type strain, thereby demonstrating a role for CopYAZ in the copper efflux of S. mutans. The complementation of the CopYAZ system restored copper expulsion, membrane potential, and stress tolerance in the copYAZ-null mutant. Taking these results collectively, we have established the function of the S. mutans CopYAZ system in copper export and have further expanded knowledge on the importance of copper homeostasis and the CopYAZ system in modulating streptococcal physiology, including stress tolerance, membrane potential, genetic competence, and biofilm formation. IMPORTANCE S. mutans is best known for its role in the initiation and progression of human dental caries, one of the most common chronic diseases worldwide. S. mutans is also implicated in bacterial endocarditis, a life-threatening inflammation of the heart valve. The core virulence factors of S. mutans include its ability to produce and sustain acidic conditions and to form a polysaccharide-encased biofilm that provides protection against environmental insults. Here, we demonstrate that the addition of copper and/or deletion of copYAZ (the copper homeostasis system) have serious implications in modulating biofilm formation, stress tolerance, and genetic transformation in S. mutans. Manipulating the pathways affected by copper and the copYAZ system may help to develop potential therapeutics to prevent S. mutans infection in and beyond the oral cavity. PMID:26013484

  2. The CopRS Two-Component System Is Responsible for Resistance to Copper in the Cyanobacterium Synechocystis sp. PCC 68031[C][W][OA

    PubMed Central

    Giner-Lamia, Joaquín; López-Maury, Luis; Reyes, José C.; Florencio, Francisco J.

    2012-01-01

    Photosynthetic organisms need copper for cytochrome oxidase and for plastocyanin in the fundamental processes of respiration and photosynthesis. However, excess of free copper is detrimental inside the cells and therefore organisms have developed homeostatic mechanisms to tightly regulate its acquisition, sequestration, and efflux. Herein we show that the CopRS two-component system (also known as Hik31-Rre34) is essential for copper resistance in Synechocystis sp. PCC 6803. It regulates expression of a putative heavy-metal efflux-resistance nodulation and division type copper efflux system (encoded by copBAC) as well as its own expression (in the copMRS operon) in response to the presence of copper in the media. Mutants in this two-component system or the efflux system render cells more sensitive to the presence of copper in the media and accumulate more intracellular copper than the wild type. Furthermore, CopS periplasmic domain is able to bind copper, suggesting that CopS could be able to detect copper directly. Both operons (copMRS and copBAC) are also induced by the photosynthetic inhibitor 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone but this induction requires the presence of copper in the media. The reduced response of two mutant strains to copper, one lacking plastocyanin and a second one impaired in copper transport to the thylakoid, due to the absence of the PI-type ATPases PacS and CtaA, suggests that CopS can detect intracellular copper. In addition, a tagged version of CopS with a triple HA epitope localizes to both the plasma and the thylakoid membranes, suggesting that CopS could be involved in copper detection in both the periplasm and the thylakoid lumen. PMID:22715108

  3. The CopRS two-component system is responsible for resistance to copper in the cyanobacterium Synechocystis sp. PCC 6803.

    PubMed

    Giner-Lamia, Joaquín; López-Maury, Luis; Reyes, José C; Florencio, Francisco J

    2012-08-01

    Photosynthetic organisms need copper for cytochrome oxidase and for plastocyanin in the fundamental processes of respiration and photosynthesis. However, excess of free copper is detrimental inside the cells and therefore organisms have developed homeostatic mechanisms to tightly regulate its acquisition, sequestration, and efflux. Herein we show that the CopRS two-component system (also known as Hik31-Rre34) is essential for copper resistance in Synechocystis sp. PCC 6803. It regulates expression of a putative heavy-metal efflux-resistance nodulation and division type copper efflux system (encoded by copBAC) as well as its own expression (in the copMRS operon) in response to the presence of copper in the media. Mutants in this two-component system or the efflux system render cells more sensitive to the presence of copper in the media and accumulate more intracellular copper than the wild type. Furthermore, CopS periplasmic domain is able to bind copper, suggesting that CopS could be able to detect copper directly. Both operons (copMRS and copBAC) are also induced by the photosynthetic inhibitor 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone but this induction requires the presence of copper in the media. The reduced response of two mutant strains to copper, one lacking plastocyanin and a second one impaired in copper transport to the thylakoid, due to the absence of the P(I)-type ATPases PacS and CtaA, suggests that CopS can detect intracellular copper. In addition, a tagged version of CopS with a triple HA epitope localizes to both the plasma and the thylakoid membranes, suggesting that CopS could be involved in copper detection in both the periplasm and the thylakoid lumen.

  4. Redox control of copper homeostasis in cyanobacteria.

    PubMed

    López-Maury, Luis; Giner-Lamia, Joaquín; Florencio, Francisco J

    2012-12-01

    Copper is essential for all living organisms but is toxic when present in excess. Therefore organisms have developed homeostatic mechanism to tightly regulate its cellular concentration. In a recent study we have shown that CopRS two-component system is essential for copper resistance in the cyanobacterium Synechocystis sp PCC 6803. This two-component regulates expression of a heavy-metal RND type copper efflux system (encoded by copBAC) as well as its own expression (in the copMRS operon) in response to an excess of copper in the media. We have also observed that both operons are induced under condition that reduces the photosynthetic electron flow and this induction depends on the presence of the copper-protein, plastocyanin. These findings, together with CopS localization to the thylakoid membrane and its periplasmic domain being able to bind copper directly, suggest that CopS could be involved in copper detection in both the periplasm and the thylakoid lumen.

  5. CopM is a novel copper-binding protein involved in copper resistance in Synechocystis sp. PCC 6803.

    PubMed

    Giner-Lamia, Joaquín; López-Maury, Luis; Florencio, Francisco J

    2015-02-01

    Copper resistance system in the cyanobacterium Synechocystis sp. PCC 6803 comprises two operons, copMRS and copBAC, which are expressed in response to copper in the media. copBAC codes for a heavy-metal efflux-resistance nodulation and division (HME-RND) system, while copMRS codes for a protein of unknown function, CopM, and a two-component system CopRS, which controls the expression of these two operons. Here, we report that CopM is a periplasmic protein able to bind Cu(I) with high affinity (KD ~3 × 10(-16) ). Mutants lacking copM showed a sensitive copper phenotype similar to mutants affected in copB, but lower than mutants of the two-component system CopRS, suggesting that CopBAC and CopM constitute two independent resistance mechanisms. Moreover, constitutive expression of copM is able to partially suppress the copper sensitivity of the copR mutant strain, pointing out that CopM per se is able to confer copper resistance. Furthermore, constitutive expression of copM was able to reduce total cellular copper content of the copR mutant to the levels determined in the wild-type (WT) strain. Finally, CopM was localized not only in the periplasm but also in the extracellular space, suggesting that CopM can also prevent copper accumulation probably by direct copper binding outside the cell. © 2014 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

  6. CopM is a novel copper-binding protein involved in copper resistance in Synechocystis sp. PCC 6803

    PubMed Central

    Giner-Lamia, Joaquín; López-Maury, Luis; Florencio, Francisco J

    2015-01-01

    Copper resistance system in the cyanobacterium Synechocystis sp. PCC 6803 comprises two operons, copMRS and copBAC, which are expressed in response to copper in the media. copBAC codes for a heavy-metal efflux–resistance nodulation and division (HME-RND) system, while copMRS codes for a protein of unknown function, CopM, and a two-component system CopRS, which controls the expression of these two operons. Here, we report that CopM is a periplasmic protein able to bind Cu(I) with high affinity (KD ∼3 × 10−16). Mutants lacking copM showed a sensitive copper phenotype similar to mutants affected in copB, but lower than mutants of the two-component system CopRS, suggesting that CopBAC and CopM constitute two independent resistance mechanisms. Moreover, constitutive expression of copM is able to partially suppress the copper sensitivity of the copR mutant strain, pointing out that CopM per se is able to confer copper resistance. Furthermore, constitutive expression of copM was able to reduce total cellular copper content of the copR mutant to the levels determined in the wild-type (WT) strain. Finally, CopM was localized not only in the periplasm but also in the extracellular space, suggesting that CopM can also prevent copper accumulation probably by direct copper binding outside the cell. PMID:25545960

  7. Streptococcus mutans copper chaperone, CopZ, is critical for biofilm formation and competitiveness.

    PubMed

    Garcia, S S; Du, Q; Wu, H

    2016-12-01

    The oral cavity is a dynamic environment characterized by hundreds of bacterial species, saliva, and an influx of nutrients and metal ions such as copper. Although there is a physiologic level of copper in the saliva, the oral cavity is often challenged with an influx of copper ions. At high concentrations copper is toxic and must therefore be strictly regulated by pathogens for them to persist and cause disease. The cariogenic pathogen Streptococcus mutans manages excess copper using the copYAZ operon that encodes a negative DNA-binding repressor (CopY), the P1-ATPase copper exporter (CopA), and the copper chaperone (CopZ). These hypothetical roles of the copYAZ operon in regulation and copper transport to receptors led us to investigate their contribution to S. mutans virulence. Mutants defective in the copper chaperone CopZ, but not CopY or CopA, were impaired in biofilm formation and competitiveness against commensal streptococci. Characterization of the CopZ mutant biofilm revealed a decreased secretion of glucosyltransferases and reduced expression of mutacin genes. These data suggest that the function of copZ on biofilm and competitiveness is independent of copper resistance and CopZ is a global regulator for biofilm and other virulence factors. Further characterization of CopZ may lead to the identification of new biofilm pathways. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  8. Global Transcriptional Profiles of the Copper Responses in the Cyanobacterium Synechocystis sp. PCC 6803

    PubMed Central

    Giner-Lamia, Joaquin; López-Maury, Luis; Florencio, Francisco J.

    2014-01-01

    Copper is an essential element involved in fundamental processes like respiration and photosynthesis. However, it becomes toxic at high concentration, which has forced organisms to control its cellular concentration. We have recently described a copper resistance system in the cyanobacterium Synechocystis sp. PCC 6803, which is mediated by the two-component system, CopRS, a RND metal transport system, CopBAC and a protein of unknown function, CopM. Here, we report the transcriptional responses to copper additions at non-toxic (0.3 µM) and toxic concentrations (3 µM) in the wild type and in the copper sensitive copR mutant strain. While 0.3 µM copper slightly stimulated metabolism and promoted the exchange between cytochrome c6 and plastocyanin as soluble electron carriers, the addition of 3 µM copper catalyzed the formation of ROS, led to a general stress response and induced expression of Fe-S cluster biogenesis genes. According to this, a double mutant strain copRsufR, which expresses constitutively the sufBCDS operon, tolerated higher copper concentration than the copR mutant strain, suggesting that Fe-S clusters are direct targets of copper toxicity in Synechocystis. In addition we have also demonstrated that InrS, a nickel binding transcriptional repressor that belong to the CsoR family of transcriptional factor, was involved in heavy metal homeostasis, including copper, in Synechocystis. Finally, global gene expression analysis of the copR mutant strain suggested that CopRS only controls the expression of copMRS and copBAC operons in response to copper. PMID:25268225

  9. Evolutionary analysis and lateral gene transfer of two-component regulatory systems associated with heavy-metal tolerance in bacteria.

    PubMed

    Bouzat, Juan L; Hoostal, Matthew J

    2013-05-01

    Microorganisms have adapted intricate signal transduction mechanisms to coordinate tolerance to toxic levels of metals, including two-component regulatory systems (TCRS). In particular, both cop and czc operons are regulated by TCRS; the cop operon plays a key role in bacterial tolerance to copper, whereas the czc operon is involved in the efflux of cadmium, zinc, and cobalt from the cell. Although the molecular physiology of heavy metal tolerance genes has been extensively studied, their evolutionary relationships are not well-understood. Phylogenetic relationships among heavy-metal efflux proteins and their corresponding two-component regulatory proteins revealed orthologous and paralogous relationships from species divergences and ancient gene duplications. The presence of heavy metal tolerance genes on bacterial plasmids suggests these genes may be prone to spread through horizontal gene transfer. Phylogenetic inferences revealed nine potential examples of lateral gene transfer associated with metal efflux proteins and two examples for regulatory proteins. Notably, four of the examples suggest lateral transfer across major evolutionary domains. In most cases, differences in GC content in metal tolerance genes and their corresponding host genomes confirmed lateral gene transfer events. Three-dimensional protein structures predicted for the response regulators encoded by cop and czc operons showed a high degree of structural similarity with other known proteins involved in TCRS signal transduction, which suggests common evolutionary origins of functional phenotypes and similar mechanisms of action for these response regulators.

  10. A copper-induced quinone degradation pathway provides protection against combined copper/quinone stress in Lactococcus lactis IL1403.

    PubMed

    Mancini, Stefano; Abicht, Helge K; Gonskikh, Yulia; Solioz, Marc

    2015-02-01

    Quinones are ubiquitous in the environment. They occur naturally but are also in widespread use in human and industrial activities. Quinones alone are relatively benign to bacteria, but in combination with copper, they become toxic by a mechanism that leads to intracellular thiol depletion. Here, it was shown that the yahCD-yaiAB operon of Lactococcus lactis IL1403 provides resistance to combined copper/quinone stress. The operon is under the control of CopR, which also regulates expression of the copRZA copper resistance operon as well as other L. lactis genes. Expression of the yahCD-yaiAB operon is induced by copper but not by quinones. Two of the proteins encoded by the operon appear to play key roles in alleviating quinone/copper stress: YaiB is a flavoprotein that converts p-benzoquinones to less toxic hydroquinones, using reduced nicotinamide adenine dinucleotide phosphate (NADPH) as reductant; YaiA is a hydroquinone dioxygenase that converts hydroquinone putatively to 4-hydroxymuconic semialdehyde in an oxygen-consuming reaction. Hydroquinone and methylhydroquinone are both substrates of YaiA. Deletion of yaiB causes increased sensitivity of L. lactis to quinones and complete growth arrest under combined quinone and copper stress. Copper induction of the yahCD-yaiAB operon offers protection to copper/quinone toxicity and could provide a growth advantage to L. lactis in some environments. © 2014 John Wiley & Sons Ltd.

  11. Response to excess copper in the hyperthermophile Sulfolobus solfataricus strain 98/2

    PubMed Central

    Villafane, Aramis; Voskoboynik, Yekaterina; Cuebas, Mariola; Ruhl, Ilona; Bini, Elisabetta

    2009-01-01

    Copper is an essential micronutrient, but toxic in excess. Sulfolobus solfataricus cells have the ability to adapt to fluctuations of copper levels in their external environment. To better understand the molecular mechanism behind the organismal response to copper, the expression of the cluster of genes copRTA, which encodes the copper-responsive transcriptional regulator CopR, the copper-binding protein CopT, and CopA, has been investigated and the whole operon has been shown to be cotranscribed at low levels from the copR promoter under all conditions, whereas increased transcription from the copTA promoter occurs in the presence of excess copper. Furthermore, the expression of the copper-transporting ATPase CopA over a 27-hour interval has been monitored by quantitative real-time RT-PCR and compared to the pattern of cellular copper accumulation, as determined in a parallel analysis by Inductively Coupled Plasma Optical Emission spectrometry (ICP-OES). The results provide the basis for a model of the molecular mechanisms of copper homeostasis in Sulfolobus, which relies on copper efflux and sequestration. PMID:19427833

  12. Successful Establishment of Plasmids R1 and pMV158 in a New Host Requires the Relief of the Transcriptional Repression of Their Essential rep Genes

    PubMed Central

    Ruiz-Masó, José Á.; Luengo, Luis M.; Moreno-Córdoba, Inmaculada; Díaz-Orejas, Ramón; del Solar, Gloria

    2017-01-01

    Although differing in size, encoded traits, host range, and replication mechanism, both narrow-host-range theta-type conjugative enterobacterial plasmid R1 and promiscuous rolling-circle-type mobilizable streptococcal plasmid pMV158 encode a transcriptional repressor protein, namely CopB in R1 and CopG in pMV158, involved in replication control. The gene encoding CopB or CopG is cotranscribed with a downstream gene that encodes the replication initiator Rep protein of the corresponding plasmid. However, whereas CopG is an auto-repressor that inhibits transcription of the entire copG-repB operon, CopB is expressed constitutively and represses a second, downstream promoter that directs transcription of repA. As a consequence of the distinct regulatory pathways implied by CopB and CopG, these repressor proteins play a different role in control of plasmid replication during the steady state: while CopB has an auxiliary role by keeping repressed the regulated promoter whenever the plasmid copy number is above a low threshold, CopG plays a primary role by acting coordinately with RNAII. Here, we have studied the role of the regulatory circuit mediated by these transcriptional repressors during the establishment of these two plasmids in a new host cell, and found that excess Cop repressor molecules in the recipient cell result in a severe decrease in the frequency and/or the velocity of appearance of transformant colonies for the cognate plasmid but not for unrelated plasmids. Using the pMV158 replicon as a model system, together with highly sensitive real-time qPCR and inverse PCR methods, we have also analyzed the effect of CopG on the kinetics of repopulation of the plasmid in Streptococcus pneumoniae. We show that, whereas in the absence of CopG pMV158 repopulation occurs mainly during the first 45 min following plasmid transfer, the presence of the transcriptional repressor in the recipient cell severely impairs the replicon repopulation and makes the plasmid replicate at approximately the same rate as the chromosome at any time after transformation, which results in maximal plasmid loss rate in the absence of selection. Overall, these findings indicate that unrepressed activity of the Cop-regulated promoter is crucial for the successful colonization of the recipient bacterial cells by the plasmid. PMID:29250051

  13. A Copper-Activated Two-Component System Interacts with Zinc and Imipenem Resistance in Pseudomonas aeruginosa▿

    PubMed Central

    Caille, Olivier; Rossier, Claude; Perron, Karl

    2007-01-01

    The effects of copper (Cu) on trace metal and antibiotic resistance of Pseudomonas aeruginosa have been investigated. Cu treatments induced resistance not only to this metal but also, surprisingly, to zinc (Zn). Quantitative reverse transcription-PCR (qRT-PCR) revealed that after Cu treatment the transcription of the czcRS two-component system (TCS) operon was enhanced as well as that of the czcCBA operon encoding an efflux pump specific for zinc, cadmium, and cobalt. Cu treatments at the same time caused a decrease in the production of OprD porin, resulting in resistance to the carbapenem antibiotic imipenem. The CzcR regulator was known to repress oprD. However, Cu was still able to decrease the production of OprD and induce imipenem resistance in a czcRS knockout mutant. This strongly suggested that another Cu-dependent regulatory system was acting negatively on oprD expression. TCS regulator genes copR-copS have been shown to be involved in Cu tolerance in P. aeruginosa. qRT-PCR showed that overproduction of the CopR or of the CzcR regulator resulted in increased transcription of the czcC gene as well as in a decrease in oprD gene transcription, either in the wild-type strain or in the czcRS knockout mutant. Overproduction experiments suggest that a metal-dependent mechanism operates at the posttranscriptional level to control the production of the CzcCBA efflux pump. This study shows that CopR is a new negative regulator of OprD porin and that it links Zn, Cu, and imipenem resistances by interacting with the CzcRS TCS. PMID:17449606

  14. A horizontally gene transferred copper resistance locus confers hyper‐resistance to antibacterial copper toxicity and enables survival of community acquired methicillin resistant Staphylococcus aureus USA300 in macrophages

    PubMed Central

    Purves, Joanne; Thomas, Jamie; Riboldi, Gustavo P.; Zapotoczna, Marta; Tarrant, Emma; Andrew, Peter W.; Londoño, Alejandra; Planet, Paul J.; Geoghegan, Joan A.; Waldron, Kevin J.

    2018-01-01

    Summary Excess copper is highly toxic and forms part of the host innate immune system's antibacterial arsenal, accumulating at sites of infection and acting within macrophages to kill engulfed pathogens. We show for the first time that a novel, horizontally gene transferred copper resistance locus (copXL), uniquely associated with the SCCmec elements of the highly virulent, epidemic, community acquired methicillin resistant Staphylococcus aureus (CA‐MRSA) USA300, confers copper hyper‐resistance. These genes are additional to existing core genome copper resistance mechanisms, and are not found in typical S. aureus lineages, but are increasingly identified in emerging pathogenic isolates. Our data show that CopX, a putative P1B‐3‐ATPase efflux transporter, and CopL, a novel lipoprotein, confer copper hyper‐resistance compared to typical S. aureus strains. The copXL genes form an operon that is tightly repressed in low copper environments by the copper regulator CsoR. Significantly, CopX and CopL are important for S. aureus USA300 intracellular survival within macrophages. Therefore, the emergence of new S. aureus clones with the copXL locus has significant implications for public health because these genes confer increased resistance to antibacterial copper toxicity, enhancing bacterial fitness by altering S. aureus interaction with innate immunity. PMID:29521441

  15. A cytosolic copper storage protein provides a second level of copper tolerance in Streptomyces lividans.

    PubMed

    Straw, Megan L; Chaplin, Amanda K; Hough, Michael A; Paps, Jordi; Bavro, Vassiliy N; Wilson, Michael T; Vijgenboom, Erik; Worrall, Jonathan A R

    2018-01-24

    Streptomyces lividans has a distinct dependence on the bioavailability of copper for its morphological development. A cytosolic copper resistance system is operative in S. lividans that serves to preclude deleterious copper levels. This system comprises of several CopZ-like copper chaperones and P 1 -type ATPases, predominantly under the transcriptional control of a metalloregulator from the copper sensitive operon repressor (CsoR) family. In the present study, we discover a new layer of cytosolic copper resistance in S. lividans that involves a protein belonging to the newly discovered family of copper storage proteins, which we have named Ccsp (cytosolic copper storage protein). From an evolutionary perspective, we find Ccsp homologues to be widespread in Bacteria and extend through into Archaea and Eukaryota. Under copper stress Ccsp is upregulated and consists of a homotetramer assembly capable of binding up to 80 cuprous ions (20 per protomer). X-ray crystallography reveals 18 cysteines, 3 histidines and 1 aspartate are involved in cuprous ion coordination. Loading of cuprous ions to Ccsp is a cooperative process with a Hill coefficient of 1.9 and a CopZ-like copper chaperone can transfer copper to Ccsp. A Δccsp mutant strain indicates that Ccsp is not required under initial copper stress in S. lividans, but as the CsoR/CopZ/ATPase efflux system becomes saturated, Ccsp facilitates a second level of copper tolerance.

  16. Copper Tolerance and Characterization of a Copper-Responsive Operon, copYAZ, in an M1T1 Clinical Strain of Streptococcus pyogenes

    PubMed Central

    Gordon, Lily D.; Fang, Zhong; Holder, Robert C.; Reid, Sean D.

    2015-01-01

    ABSTRACT Infection with Streptococcus pyogenes is associated with a breadth of clinical manifestations ranging from mild pharyngitis to severe necrotizing fasciitis. Elevated levels of intracellular copper are highly toxic to this bacterium, and thus, the microbe must tightly regulate the level of this metal ion by one or more mechanisms, which have, to date, not been clearly defined. In this study, we have identified two virulence mechanisms by which S. pyogenes protects itself against copper toxicity. We defined a set of putative genes, copY (for a regulator), copA (for a P1-type ATPase), and copZ (for a copper chaperone), whose expression is regulated by copper. Our results indicate that these genes are highly conserved among a range of clinical S. pyogenes isolates. The copY, copA, and copZ genes are induced by copper and are transcribed as a single unit. Heterologous expression assays revealed that S. pyogenes CopA can confer copper tolerance in a copper-sensitive Escherichia coli mutant by preventing the accumulation of toxic levels of copper, a finding that is consistent with a role for CopA in copper export. Evaluation of the effect of copper stress on S. pyogenes in a planktonic or biofilm state revealed that biofilms may aid in protection during initial exposure to copper. However, copper stress appears to prevent the shift from the planktonic to the biofilm state. Therefore, our results indicate that S. pyogenes may use several virulence mechanisms, including altered gene expression and a transition to and from planktonic and biofilm states, to promote survival during copper stress. IMPORTANCE Bacterial pathogens encounter multiple stressors at the host-pathogen interface. This study evaluates a virulence mechanism(s) utilized by S. pyogenes to combat copper at sites of infection. A better understanding of pathogen tolerance to stressors such as copper is necessary to determine how host-pathogen interactions impact bacterial survival during infections. These insights may lead to the identification of novel therapeutic targets that can be used to address antibiotic resistance. PMID:26013489

  17. Copper Tolerance and Characterization of a Copper-Responsive Operon, copYAZ, in an M1T1 Clinical Strain of Streptococcus pyogenes.

    PubMed

    Young, Christie A; Gordon, Lily D; Fang, Zhong; Holder, Robert C; Reid, Sean D

    2015-08-01

    Infection with Streptococcus pyogenes is associated with a breadth of clinical manifestations ranging from mild pharyngitis to severe necrotizing fasciitis. Elevated levels of intracellular copper are highly toxic to this bacterium, and thus, the microbe must tightly regulate the level of this metal ion by one or more mechanisms, which have, to date, not been clearly defined. In this study, we have identified two virulence mechanisms by which S. pyogenes protects itself against copper toxicity. We defined a set of putative genes, copY (for a regulator), copA (for a P1-type ATPase), and copZ (for a copper chaperone), whose expression is regulated by copper. Our results indicate that these genes are highly conserved among a range of clinical S. pyogenes isolates. The copY, copA, and copZ genes are induced by copper and are transcribed as a single unit. Heterologous expression assays revealed that S. pyogenes CopA can confer copper tolerance in a copper-sensitive Escherichia coli mutant by preventing the accumulation of toxic levels of copper, a finding that is consistent with a role for CopA in copper export. Evaluation of the effect of copper stress on S. pyogenes in a planktonic or biofilm state revealed that biofilms may aid in protection during initial exposure to copper. However, copper stress appears to prevent the shift from the planktonic to the biofilm state. Therefore, our results indicate that S. pyogenes may use several virulence mechanisms, including altered gene expression and a transition to and from planktonic and biofilm states, to promote survival during copper stress. Bacterial pathogens encounter multiple stressors at the host-pathogen interface. This study evaluates a virulence mechanism(s) utilized by S. pyogenes to combat copper at sites of infection. A better understanding of pathogen tolerance to stressors such as copper is necessary to determine how host-pathogen interactions impact bacterial survival during infections. These insights may lead to the identification of novel therapeutic targets that can be used to address antibiotic resistance. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  18. Control of copper resistance and inorganic sulfur metabolism by paralogous regulators in Staphylococcus aureus.

    PubMed

    Grossoehme, Nicholas; Kehl-Fie, Thomas E; Ma, Zhen; Adams, Keith W; Cowart, Darin M; Scott, Robert A; Skaar, Eric P; Giedroc, David P

    2011-04-15

    All strains of Staphylococcus aureus encode a putative copper-sensitive operon repressor (CsoR) and one other CsoR-like protein of unknown function. We show here that NWMN_1991 encodes a bona fide Cu(I)-inducible CsoR of a genetically unlinked copA-copZ copper resistance operon in S. aureus strain Newman. In contrast, an unannotated open reading frame found between NWMN_0027 and NWMN_0026 (denoted NWMN_0026.5) encodes a CsoR-like regulator that represses expression of adjacent genes by binding specifically to a pair of canonical operator sites positioned in the NWMN_0027-0026.5 intergenic region. Inspection of these regulated genes suggests a role in assimilation of inorganic sulfur from thiosulfate and vectorial sulfur transfer, and we designate NWMN_0026.5 as CstR (CsoR-like sulfur transferase repressor). Expression analysis demonstrates that CsoR and CstR control their respective regulons in response to distinct stimuli with no overlap in vivo. Unlike CsoR, CstR does not form a stable complex with Cu(I); operator binding is instead inhibited by oxidation of the intersubunit cysteine pair to a mixture of disulfide and trisulfide linkages by a likely metabolite of thiosulfate assimilation, sulfite. CsoR is unreactive toward sulfite under the same conditions. We conclude that CsoR and CstR are paralogs in S. aureus that function in the same cytoplasm to control distinct physiological processes.

  19. Control of Copper Resistance and Inorganic Sulfur Metabolism by Paralogous Regulators in Staphylococcus aureus*

    PubMed Central

    Grossoehme, Nicholas; Kehl-Fie, Thomas E.; Ma, Zhen; Adams, Keith W.; Cowart, Darin M.; Scott, Robert A.; Skaar, Eric P.; Giedroc, David P.

    2011-01-01

    All strains of Staphylococcus aureus encode a putative copper-sensitive operon repressor (CsoR) and one other CsoR-like protein of unknown function. We show here that NWMN_1991 encodes a bona fide Cu(I)-inducible CsoR of a genetically unlinked copA-copZ copper resistance operon in S. aureus strain Newman. In contrast, an unannotated open reading frame found between NWMN_0027 and NWMN_0026 (denoted NWMN_0026.5) encodes a CsoR-like regulator that represses expression of adjacent genes by binding specifically to a pair of canonical operator sites positioned in the NWMN_0027–0026.5 intergenic region. Inspection of these regulated genes suggests a role in assimilation of inorganic sulfur from thiosulfate and vectorial sulfur transfer, and we designate NWMN_0026.5 as CstR (CsoR-like sulfur transferase repressor). Expression analysis demonstrates that CsoR and CstR control their respective regulons in response to distinct stimuli with no overlap in vivo. Unlike CsoR, CstR does not form a stable complex with Cu(I); operator binding is instead inhibited by oxidation of the intersubunit cysteine pair to a mixture of disulfide and trisulfide linkages by a likely metabolite of thiosulfate assimilation, sulfite. CsoR is unreactive toward sulfite under the same conditions. We conclude that CsoR and CstR are paralogs in S. aureus that function in the same cytoplasm to control distinct physiological processes. PMID:21339296

  20. Selfish operons: the evolutionary impact of gene clustering in prokaryotes and eukaryotes.

    PubMed

    Lawrence, J

    1999-12-01

    The Selfish Operon Model postulates that the organization of bacterial genes into operons is beneficial to the constituent genes in that proximity allows horizontal cotransfer of all genes required for a selectable phenotype; eukaryotic operons formed for very different reasons. Horizontal transfer of selfish operons most probably promotes bacterial diversification.

  1. 20 CFR 10.223 - Are there other circumstances under which OWCP will not authorize payment of COP?

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... will not authorize payment of COP? 10.223 Section 10.223 Employees' Benefits OFFICE OF WORKERS... of Cop § 10.223 Are there other circumstances under which OWCP will not authorize payment of COP... examination required by OWCP, the right to COP is suspended until the refusal or obstruction ceases. COP...

  2. 20 CFR 10.223 - Are there other circumstances under which OWCP will not authorize payment of COP?

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... OWCP will not authorize payment of COP? 10.223 Section 10.223 Employees' Benefits OFFICE OF WORKERS... of Cop § 10.223 Are there other circumstances under which OWCP will not authorize payment of COP... examination required by OWCP, the right to COP is suspended until the refusal or obstruction ceases. COP...

  3. 20 CFR 10.223 - Are there other circumstances under which OWCP will not authorize payment of COP?

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... OWCP will not authorize payment of COP? 10.223 Section 10.223 Employees' Benefits OFFICE OF WORKERS... of Cop § 10.223 Are there other circumstances under which OWCP will not authorize payment of COP... examination required by OWCP, the right to COP is suspended until the refusal or obstruction ceases. COP...

  4. 20 CFR 10.223 - Are there other circumstances under which OWCP will not authorize payment of COP?

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... will not authorize payment of COP? 10.223 Section 10.223 Employees' Benefits OFFICE OF WORKERS... of Cop § 10.223 Are there other circumstances under which OWCP will not authorize payment of COP... examination required by OWCP, the right to COP is suspended until the refusal or obstruction ceases. COP...

  5. 20 CFR 10.223 - Are there other circumstances under which OWCP will not authorize payment of COP?

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... OWCP will not authorize payment of COP? 10.223 Section 10.223 Employees' Benefits OFFICE OF WORKERS... of Cop § 10.223 Are there other circumstances under which OWCP will not authorize payment of COP... examination required by OWCP, the right to COP is suspended until the refusal or obstruction ceases. COP...

  6. Metal Resistance and Lithoautotrophy in the Extreme Thermoacidophile Metallosphaera sedula

    PubMed Central

    Maezato, Yukari; Johnson, Tyler; McCarthy, Samuel; Dana, Karl

    2012-01-01

    Archaea such as Metallosphaera sedula are thermophilic lithoautotrophs that occupy unusually acidic and metal-rich environments. These traits are thought to underlie their industrial importance for bioleaching of base and precious metals. In this study, a genetic approach was taken to investigate the specific relationship between metal resistance and lithoautotrophy during biotransformation of the primary copper ore, chalcopyrite (CuFeS2). In this study, a genetic system was developed for M. sedula to investigate parameters that limit bioleaching of chalcopyrite. The functional role of the M. sedula copRTA operon was demonstrated by cross-species complementation of a copper-sensitive Sulfolobus solfataricus copR mutant. Inactivation of the gene encoding the M. sedula copper efflux protein, copA, using targeted recombination compromised metal resistance and eliminated chalcopyrite bioleaching. In contrast, a spontaneous M. sedula mutant (CuR1) with elevated metal resistance transformed chalcopyrite at an accelerated rate without affecting chemoheterotrophic growth. Proteomic analysis of CuR1 identified pleiotropic changes, including altered abundance of transport proteins having AAA-ATPase motifs. Addition of the insoluble carbonate mineral witherite (BaCO3) further stimulated chalcopyrite lithotrophy, indicating that carbon was a limiting factor. Since both mineral types were actively colonized, enhanced metal leaching may arise from the cooperative exchange of energy and carbon between surface-adhered populations. Genetic approaches provide a new means of improving the efficiency of metal bioleaching by enhancing the mechanistic understanding of thermophilic lithoautotrophy. PMID:23065978

  7. Teaching the Big Ideas of Biology with Operon Models

    ERIC Educational Resources Information Center

    Cooper, Robert A.

    2015-01-01

    This paper presents an activity that engages students in model-based reasoning, requiring them to predict the behavior of the trp and lac operons under different environmental conditions. Students are presented six scenarios for the "trp" operon and five for the "lac" operon. In most of the scenarios, specific mutations have…

  8. 30 CFR 585.631 - When must I initiate activities under an approved COP?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... approved COP? 585.631 Section 585.631 Mineral Resources BUREAU OF OCEAN ENERGY MANAGEMENT, DEPARTMENT OF... CONTINENTAL SHELF Plans and Information Requirements Activities Under An Approved Cop § 585.631 When must I initiate activities under an approved COP? After your COP is approved, you must commence construction by...

  9. 30 CFR 285.631 - When must I initiate activities under an approved COP?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... approved COP? 285.631 Section 285.631 Mineral Resources MINERALS MANAGEMENT SERVICE, DEPARTMENT OF THE... Plans and Information Requirements Activities Under An Approved Cop § 285.631 When must I initiate activities under an approved COP? After your COP is approved, you must commence construction by the date...

  10. 30 CFR 285.631 - When must I initiate activities under an approved COP?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... approved COP? 285.631 Section 285.631 Mineral Resources BUREAU OF OCEAN ENERGY MANAGEMENT, REGULATION, AND... OUTER CONTINENTAL SHELF Plans and Information Requirements Activities Under An Approved Cop § 285.631 When must I initiate activities under an approved COP? After your COP is approved, you must commence...

  11. 30 CFR 585.631 - When must I initiate activities under an approved COP?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... approved COP? 585.631 Section 585.631 Mineral Resources BUREAU OF OCEAN ENERGY MANAGEMENT, DEPARTMENT OF... CONTINENTAL SHELF Plans and Information Requirements Activities Under An Approved Cop § 585.631 When must I initiate activities under an approved COP? After your COP is approved, you must commence construction by...

  12. 30 CFR 585.631 - When must I initiate activities under an approved COP?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... approved COP? 585.631 Section 585.631 Mineral Resources BUREAU OF OCEAN ENERGY MANAGEMENT, DEPARTMENT OF... CONTINENTAL SHELF Plans and Information Requirements Activities Under An Approved Cop § 585.631 When must I initiate activities under an approved COP? After your COP is approved, you must commence construction by...

  13. 20 CFR 10.200 - What is continuation of pay?

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ..., or COP. The employer, not OWCP, pays COP. Unlike wage loss benefits, COP is subject to taxes and all... (c) of this section, who is eligible for COP, and may not require the employee to use his or her own... continuing the employee's pay, the employer may controvert the employee's COP entitlement pending a final...

  14. 20 CFR 10.200 - What is continuation of pay?

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ..., or COP. The employer, not OWCP, pays COP. Unlike wage loss benefits, COP is subject to taxes and all... employee who is eligible for COP, and may not require the employee to use his or her own sick or annual... employee's pay, the employer may controvert the employee's COP entitlement pending a final determination by...

  15. 20 CFR 10.200 - What is continuation of pay?

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ..., or COP. The employer, not OWCP, pays COP. Unlike wage loss benefits, COP is subject to taxes and all... (c) of this section, who is eligible for COP, and may not require the employee to use his or her own... continuing the employee's pay, the employer may controvert the employee's COP entitlement pending a final...

  16. 20 CFR 10.200 - What is continuation of pay?

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ..., or COP. The employer, not OWCP, pays COP. Unlike wage loss benefits, COP is subject to taxes and all... employee who is eligible for COP, and may not require the employee to use his or her own sick or annual... employee's pay, the employer may controvert the employee's COP entitlement pending a final determination by...

  17. 20 CFR 10.200 - What is continuation of pay?

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ..., or COP. The employer, not OWCP, pays COP. Unlike wage loss benefits, COP is subject to taxes and all... (c) of this section, who is eligible for COP, and may not require the employee to use his or her own... continuing the employee's pay, the employer may controvert the employee's COP entitlement pending a final...

  18. Operon Formation is Driven by Co-Regulation and Not by Horizontal Gene Transfer

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

    Price, Morgan N.; Huang, Katherine H.; Arkin, Adam P.

    Although operons are often subject to horizontal gene transfer (HGT), non-HGT genes are particularly likely to be in operons. To resolve this apparent discrepancy and to determine whether HGT is involved in operon formation, we examined the evolutionary history of the genes and operons in Escherichia coli K12. We show that genes that have homologs in distantly related bacteria but not in close relatives of E. coli (indicating HGTi) form new operons at about the same rates as native genes. Furthermore, genes in new operons are no more likely than other genes to have phylogenetic trees that are inconsistent withmore » the species tree. In contrast, essential genes and ubiquitous genes without paralogs (genes believed to undergo HGT rarely) often form new operons. We conclude that HGT is not associated with operon formation, but instead promotes the prevalence of pre-existing operons. To explain operon formation, we propose that new operons reduce the amount of regulatory information required to specify optimal expression patterns. Consistent with this hypothesis, operons have greater amounts of conserved regulatory sequences than do individually transcribed genes.« less

  19. 30 CFR 585.659 - What requirements must I include in my SAP, COP, or GAP regarding air quality?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 30 Mineral Resources 2 2013-07-01 2013-07-01 false What requirements must I include in my SAP, COP, or GAP regarding air quality? 585.659 Section 585.659 Mineral Resources BUREAU OF OCEAN ENERGY... What requirements must I include in my SAP, COP, or GAP regarding air quality? (a) You must comply with...

  20. 30 CFR 585.659 - What requirements must I include in my SAP, COP, or GAP regarding air quality?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 30 Mineral Resources 2 2014-07-01 2014-07-01 false What requirements must I include in my SAP, COP, or GAP regarding air quality? 585.659 Section 585.659 Mineral Resources BUREAU OF OCEAN ENERGY... What requirements must I include in my SAP, COP, or GAP regarding air quality? (a) You must comply with...

  1. 30 CFR 585.659 - What requirements must I include in my SAP, COP, or GAP regarding air quality?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 30 Mineral Resources 2 2012-07-01 2012-07-01 false What requirements must I include in my SAP, COP, or GAP regarding air quality? 585.659 Section 585.659 Mineral Resources BUREAU OF OCEAN ENERGY... What requirements must I include in my SAP, COP, or GAP regarding air quality? (a) You must comply with...

  2. 30 CFR 285.620 - What is a Construction and Operations Plan (COP)?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 30 Mineral Resources 2 2010-07-01 2010-07-01 false What is a Construction and Operations Plan (COP... Information Requirements Construction and Operations Plan for Commercial Leases § 285.620 What is a Construction and Operations Plan (COP)? The COP describes your construction, operations, and conceptual...

  3. 30 CFR 285.635 - What must I do if I cease activities approved in my COP before the end of my commercial lease?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... in my COP before the end of my commercial lease? 285.635 Section 285.635 Mineral Resources BUREAU OF... Requirements Activities Under An Approved Cop § 285.635 What must I do if I cease activities approved in my COP... cease commercial operations, without an approved suspension, under your approved COP. If you cease...

  4. 30 CFR 285.618 - What must I do upon completion of approved site assessment activities?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... expiration of your site assessment term, you timely submit a COP meeting the requirements of this subpart, or... your COP for approval or FERC reviews your license application for approval. (b) You are not required... your approved COP or approved FERC license. (c) If, following the technical and environmental review of...

  5. 30 CFR 285.636 - What notices must I provide MMS following approval of my COP?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... approval of my COP? 285.636 Section 285.636 Mineral Resources MINERALS MANAGEMENT SERVICE, DEPARTMENT OF... SHELF Plans and Information Requirements Activities Under An Approved Cop § 285.636 What notices must I provide MMS following approval of my COP? You must notify MMS in writing of the following events, within...

  6. 30 CFR 585.636 - What notices must I provide BOEM following approval of my COP?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... approval of my COP? 585.636 Section 585.636 Mineral Resources BUREAU OF OCEAN ENERGY MANAGEMENT, DEPARTMENT... CONTINENTAL SHELF Plans and Information Requirements Activities Under An Approved Cop § 585.636 What notices must I provide BOEM following approval of my COP? You must notify BOEM in writing of the following...

  7. 30 CFR 285.636 - What notices must I provide MMS following approval of my COP?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... approval of my COP? 285.636 Section 285.636 Mineral Resources BUREAU OF OCEAN ENERGY MANAGEMENT, REGULATION... FACILITIES ON THE OUTER CONTINENTAL SHELF Plans and Information Requirements Activities Under An Approved Cop § 285.636 What notices must I provide MMS following approval of my COP? You must notify MMS in writing...

  8. 30 CFR 585.636 - What notices must I provide BOEM following approval of my COP?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... approval of my COP? 585.636 Section 585.636 Mineral Resources BUREAU OF OCEAN ENERGY MANAGEMENT, DEPARTMENT... CONTINENTAL SHELF Plans and Information Requirements Activities Under An Approved Cop § 585.636 What notices must I provide BOEM following approval of my COP? You must notify BOEM in writing of the following...

  9. 30 CFR 585.636 - What notices must I provide BOEM following approval of my COP?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... approval of my COP? 585.636 Section 585.636 Mineral Resources BUREAU OF OCEAN ENERGY MANAGEMENT, DEPARTMENT... CONTINENTAL SHELF Plans and Information Requirements Activities Under An Approved Cop § 585.636 What notices must I provide BOEM following approval of my COP? You must notify BOEM in writing of the following...

  10. 30 CFR 285.815 - What must I do if I have facility damage or an equipment failure?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... Assessments for Activities Conducted Under SAPs, COPs and GAPs Equipment Failure and Adverse Environmental... damage or failure under § 285.831, MMS may require you to revise your SAP, COP, or GAP to describe how you will address the facility damage or failure as required by § 285.634 (COP), § 285.617 (SAP), § 285...

  11. 30 CFR 585.902 - What are the general requirements for decommissioning for facilities authorized under my SAP, COP...

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... decommissioning for facilities authorized under my SAP, COP, or GAP? 585.902 Section 585.902 Mineral Resources... authorized under my SAP, COP, or GAP? (a) Except as otherwise authorized by BOEM under § 585.909, within 2... decommissioning the facilities under your SAP, COP, or GAP, you must submit a decommissioning application and...

  12. 30 CFR 585.902 - What are the general requirements for decommissioning for facilities authorized under my SAP, COP...

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... decommissioning for facilities authorized under my SAP, COP, or GAP? 585.902 Section 585.902 Mineral Resources..., Inspections, and Facility Assessments for Activities Conducted Under SAPs, COPs and GAPs Decommissioning... authorized under my SAP, COP, or GAP? (a) Except as otherwise authorized by BOEM under § 585.909, within 2...

  13. 30 CFR 585.902 - What are the general requirements for decommissioning for facilities authorized under my SAP, COP...

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... decommissioning for facilities authorized under my SAP, COP, or GAP? 585.902 Section 585.902 Mineral Resources... authorized under my SAP, COP, or GAP? (a) Except as otherwise authorized by BOEM under § 585.909, within 2... decommissioning the facilities under your SAP, COP, or GAP, you must submit a decommissioning application and...

  14. 30 CFR 285.902 - What are the general requirements for decommissioning for facilities authorized under my SAP, COP...

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... decommissioning for facilities authorized under my SAP, COP, or GAP? 285.902 Section 285.902 Mineral Resources... SAP, COP, or GAP? (a) Except as otherwise authorized by MMS under § 285.909, within 2 years following... under your SAP, COP, or GAP, you must submit a decommissioning application and receive approval from the...

  15. Genetic basis and importance of metal resistant genes in bacteria for bioremediation of contaminated environments with toxic metal pollutants.

    PubMed

    Das, Surajit; Dash, Hirak R; Chakraborty, Jaya

    2016-04-01

    Metal pollution is one of the most persistent and complex environmental issues, causing threat to the ecosystem and human health. On exposure to several toxic metals such as arsenic, cadmium, chromium, copper, lead, and mercury, several bacteria has evolved with many metal-resistant genes as a means of their adaptation. These genes can be further exploited for bioremediation of the metal-contaminated environments. Many operon-clustered metal-resistant genes such as cadB, chrA, copAB, pbrA, merA, and NiCoT have been reported in bacterial systems for cadmium, chromium, copper, lead, mercury, and nickel resistance and detoxification, respectively. The field of environmental bioremediation has been ameliorated by exploiting diverse bacterial detoxification genes. Genetic engineering integrated with bioremediation assists in manipulation of bacterial genome which can enhance toxic metal detoxification that is not usually performed by normal bacteria. These techniques include genetic engineering with single genes or operons, pathway construction, and alternations of the sequences of existing genes. However, numerous facets of bacterial novel metal-resistant genes are yet to be explored for application in microbial bioremediation practices. This review describes the role of bacteria and their adaptive mechanisms for toxic metal detoxification and restoration of contaminated sites.

  16. COP1 is required for UV-B–induced nuclear accumulation of the UVR8 photoreceptor

    PubMed Central

    Skvortsova, Mariya Y.; Loubéry, Sylvain

    2016-01-01

    The UV-B photoreceptor UV RESISTANCE LOCUS 8 (UVR8) promotes UV-B acclimation and tolerance in Arabidopsis thaliana. UVR8 localizes to both cytosol and nucleus, but its main activity is assumed to be nuclear. UV-B photoreception stimulates nuclear accumulation of UVR8 in a presently unknown manner. Here, we show that CONSTITUTIVELY PHOTOMORPHOGENIC 1 (COP1) is required for UV-B–induced nuclear accumulation of UVR8, but bypassing the COP1 requirement for UVR8 nuclear accumulation did not rescue the cop1 mutant UV-B phenotype. Using a glucocorticoid receptor (GR)-based fusion protein system to conditionally localize GR-UVR8 to the nucleus, we have demonstrated that both photoactivation and nuclear localization of UVR8 are required for UV-B–induced photomorphogenic responses. In contrast, there was no UV-B response when UV-B–activated UVR8 was artificially retained in the cytosol. In agreement with a predominantly nuclear activity, constitutively active UVR8W285A accumulated in the nucleus also in the absence of UV-B. Furthermore, GR-COP1 expression lines suggested that UV-B–activated UVR8 can be coimported into the nucleus by COP1. Our data strongly support localization of UVR8 signaling in the nucleus and a dual role for COP1 in the regulation of UV-B–induced UVR8 nuclear accumulation and in UVR8-mediated UV-B signaling. PMID:27407149

  17. The blue light-induced interaction of cryptochrome 1 with COP1 requires SPA proteins during Arabidopsis light signaling.

    PubMed

    Holtkotte, Xu; Ponnu, Jathish; Ahmad, Margaret; Hoecker, Ute

    2017-10-01

    Plants constantly adjust their growth, development and metabolism to the ambient light environment. Blue light is sensed by the Arabidopsis photoreceptors CRY1 and CRY2 which subsequently initiate light signal transduction by repressing the COP1/SPA E3 ubiquitin ligase. While the interaction between cryptochromes and SPA is blue light-dependent, it was proposed that CRY1 interacts with COP1 constitutively, i.e. also in darkness. Here, our in vivo co-immunoprecipitation experiments suggest that CRY1 and CRY2 form a complex with COP1 only after seedlings were exposed to blue light. No association between COP1 and CRY1 or CRY2 was observed in dark-grown seedlings. Thus, our results suggest that cryptochromes bind the COP1/SPA complex after photoactivation by blue light. In a spa quadruple mutant that is devoid of all four SPA proteins, CRY1 and COP1 did not interact in vivo, neither in dark-grown nor in blue light-grown seedlings. Hence, SPA proteins are required for the high-affinity interaction between CRY1 and COP1 in blue light. Yeast three-hybrid experiments also show that SPA1 enhances the CRY1-COP1 interaction. The coiled-coil domain of SPA1 which is responsible for COP1-binding was necessary to mediate a CRY1-SPA1 interaction in vivo, implying that-in turn-COP1 may be necessary for a CRY1-SPA1 complex formation. Hence, SPA1 and COP1 may act cooperatively in recognizing and binding photoactivated CRY1. In contrast, the blue light-induced association between CRY2 and COP1 was not dependent on SPA proteins in vivo. Similarly, ΔCC-SPA1 interacted with CRY2, though with a much lower affinity than wild-type SPA1. In total, our results demonstrate that CRY1 and CRY2 strongly differ in their blue light-induced interaction with the COP1/SPA complex.

  18. 30 CFR 585.815 - What must I do if I have facility damage or an equipment failure?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... Conducted Under SAPs, COPs and GAPs Equipment Failure and Adverse Environmental Effects § 585.815 What must... failure under § 585.831, BOEM may require you to revise your SAP, COP, or GAP to describe how you will address the facility damage or failure as required by § 585.634 (COP), § 585.617 (SAP), § 585.655 (GAP...

  19. 30 CFR 585.815 - What must I do if I have facility damage or an equipment failure?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... Conducted Under SAPs, COPs and GAPs Equipment Failure and Adverse Environmental Effects § 585.815 What must... failure under § 585.831, BOEM may require you to revise your SAP, COP, or GAP to describe how you will address the facility damage or failure as required by § 585.634 (COP), § 585.617 (SAP), § 585.655 (GAP...

  20. 30 CFR 585.815 - What must I do if I have facility damage or an equipment failure?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... Conducted Under SAPs, COPs and GAPs Equipment Failure and Adverse Environmental Effects § 585.815 What must... failure under § 585.831, BOEM may require you to revise your SAP, COP, or GAP to describe how you will address the facility damage or failure as required by § 585.634 (COP), § 585.617 (SAP), § 585.655 (GAP...

  1. 30 CFR 285.638 - What must I do upon completion of my commercial operations as approved in my COP or FERC license?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... operations as approved in my COP or FERC license? 285.638 Section 285.638 Mineral Resources MINERALS... FACILITIES ON THE OUTER CONTINENTAL SHELF Plans and Information Requirements Activities Under An Approved Cop § 285.638 What must I do upon completion of my commercial operations as approved in my COP or FERC...

  2. 30 CFR 285.632 - What documents must I submit before I may construct and install facilities under my approved COP?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... construct and install facilities under my approved COP? 285.632 Section 285.632 Mineral Resources BUREAU OF... Requirements Activities Under An Approved Cop § 285.632 What documents must I submit before I may construct and install facilities under my approved COP? (a) You must submit to MMS the documents listed in the following...

  3. 30 CFR 285.632 - What documents must I submit before I may construct and install facilities under my approved COP?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... construct and install facilities under my approved COP? 285.632 Section 285.632 Mineral Resources MINERALS... FACILITIES ON THE OUTER CONTINENTAL SHELF Plans and Information Requirements Activities Under An Approved Cop... COP? (a) You must submit to MMS the documents listed in the following table: ER29AP09.124 (b) You must...

  4. 30 CFR 285.700 - What reports must I submit to MMS before installing facilities described in my approved SAP, COP...

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... installing facilities described in my approved SAP, COP, or GAP? 285.700 Section 285.700 Mineral Resources... described in my approved SAP, COP, or GAP? (a) You must submit the following reports to MMS before installing facilities described in your approved COP (§ 285.632(a)) and, when required by this part, your SAP...

  5. 30 CFR 585.700 - What reports must I submit to BOEM before installing facilities described in my approved SAP, COP...

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... installing facilities described in my approved SAP, COP, or GAP? 585.700 Section 585.700 Mineral Resources... approved SAP, COP, or GAP? (a) You must submit the following reports to BOEM before installing facilities described in your approved COP (§ 585.632(a)) and, when required by this part, your SAP (§ 585.614(b)) or...

  6. 30 CFR 585.638 - What must I do upon completion of my commercial operations as approved in my COP or FERC license?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... operations as approved in my COP or FERC license? 585.638 Section 585.638 Mineral Resources BUREAU OF OCEAN... FACILITIES ON THE OUTER CONTINENTAL SHELF Plans and Information Requirements Activities Under An Approved Cop § 585.638 What must I do upon completion of my commercial operations as approved in my COP or FERC...

  7. 30 CFR 585.638 - What must I do upon completion of my commercial operations as approved in my COP or FERC license?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... operations as approved in my COP or FERC license? 585.638 Section 585.638 Mineral Resources BUREAU OF OCEAN... FACILITIES ON THE OUTER CONTINENTAL SHELF Plans and Information Requirements Activities Under An Approved Cop § 585.638 What must I do upon completion of my commercial operations as approved in my COP or FERC...

  8. 30 CFR 585.700 - What reports must I submit to BOEM before installing facilities described in my approved SAP, COP...

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... installing facilities described in my approved SAP, COP, or GAP? 585.700 Section 585.700 Mineral Resources... approved SAP, COP, or GAP? (a) You must submit the following reports to BOEM before installing facilities described in your approved COP (§ 585.632(a)) and, when required by this part, your SAP (§ 585.614(b)) or...

  9. 30 CFR 585.700 - What reports must I submit to BOEM before installing facilities described in my approved SAP, COP...

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... installing facilities described in my approved SAP, COP, or GAP? 585.700 Section 585.700 Mineral Resources... approved SAP, COP, or GAP? (a) You must submit the following reports to BOEM before installing facilities described in your approved COP (§ 585.632(a)) and, when required by this part, your SAP (§ 585.614(b)) or...

  10. 30 CFR 585.638 - What must I do upon completion of my commercial operations as approved in my COP or FERC license?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... operations as approved in my COP or FERC license? 585.638 Section 585.638 Mineral Resources BUREAU OF OCEAN... FACILITIES ON THE OUTER CONTINENTAL SHELF Plans and Information Requirements Activities Under An Approved Cop § 585.638 What must I do upon completion of my commercial operations as approved in my COP or FERC...

  11. Comprehensive genomic and phenotypic metal resistance profile of Pseudomonas putida strain S13.1.2 isolated from a vineyard soil.

    PubMed

    Chong, Teik Min; Yin, Wai-Fong; Chen, Jian-Woon; Mondy, Samuel; Grandclément, Catherine; Faure, Denis; Dessaux, Yves; Chan, Kok-Gan

    2016-12-01

    Trace metals are required in many cellular processes in bacteria but also induce toxic effects to cells when present in excess. As such, various forms of adaptive responses towards extracellular trace metal ions are essential for the survival and fitness of bacteria in their environment. A soil Pseudomonas putida, strain S13.1.2 has been isolated from French vineyard soil samples, and shown to confer resistance to copper ions. Further investigation revealed a high capacity to tolerate elevated concentrations of various heavy metals including nickel, cobalt, cadmium, zinc and arsenic. The complete genome analysis was conducted using single-molecule real-time (SMRT) sequencing and the genome consisted in a single chromosome at the size of 6.6 Mb. Presence of operons and gene clusters such as cop, cus, czc, nik, and asc systems were detected and accounted for the observed resistance phenotypes. The unique features in terms of specificity and arrangements of some genetic determinants were also highlighted in the study. Our findings has provided insights into the adaptation of this strain to accumulation and persistence of copper and other heavy metals in vineyard soil environment.

  12. 30 CFR 585.907 - How will BOEM process my decommissioning application?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... decommissioning application with the decommissioning general concept in your approved SAP, COP, or GAP to..., COP, or GAP, and BOEM will begin the appropriate NEPA analysis and other regulatory reviews as... change in the impacts previously identified and evaluated in your SAP, COP, or GAP; (2) Require any...

  13. 30 CFR 585.907 - How will BOEM process my decommissioning application?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... decommissioning application with the decommissioning general concept in your approved SAP, COP, or GAP to..., COP, or GAP, and BOEM will begin the appropriate NEPA analysis and other regulatory reviews as... change in the impacts previously identified and evaluated in your SAP, COP, or GAP; (2) Require any...

  14. CopZ from Bacillus subtilis interacts in vivo with a copper exporting CPx-type ATPase CopA.

    PubMed

    Radford, David S; Kihlken, Margaret A; Borrelly, Gilles P M; Harwood, Colin R; Le Brun, Nick E; Cavet, Jennifer S

    2003-03-14

    The structure of the hypothetical copper-metallochaperone CopZ from Bacillus subtilis and its predicted partner CopA have been studied but their respective contributions to copper export, -import, -sequestration and -supply are unknown. DeltacopA was hypersensitive to copper and contained more copper atoms cell(-1) than wild-type. Expression from the copA operator-promoter increased in elevated copper (not other metals), consistent with a role in copper export. A bacterial two-hybrid assay revealed in vivo interaction between CopZ and the N-terminal domain of CopA but not that of a related transporter, YvgW, involved in cadmium-resistance. Activity of copper-requiring cytochrome caa(3) oxidase was retained in deltacopZ and deltacopA. DeltacopZ was only slightly copper-hypersensitive but deltacopZ/deltacopA was more sensitive than deltacopA, implying some action of CopZ that is independent of CopA. Significantly, deltacopZ contained fewer copper atoms cell(-1) than wild-type under these conditions. CopZ makes a net contribution to copper sequestration and/or recycling exceeding any donation to CopA for export.

  15. Roles of Salmonella typhimurium umuDC and samAB in UV mutagenesis and UV sensitivity.

    PubMed Central

    Nohmi, T; Yamada, M; Watanabe, M; Murayama, S Y; Sofuni, T

    1992-01-01

    Expression of the umuDC operon is required for UV mutagenesis and most chemical mutagenesis in Escherichia coli. The closely related species Salmonella typhimurium has two sets of umuDC-like operons; the samAB operon is located in a 60-MDa cryptic plasmid, while the S. typhimurium umuDC (umuDCST) operon resides in a chromosome. The roles of these two umuDC-like operons in UV mutagenesis and UV sensitivity of S. typhimurium were investigated. A pBR322-derived plasmid carrying the samAB operon more efficiently restored UV mutability to a umuD44 strain and a umuC122::Tn5 strain of E. coli than a plasmid carrying the umuDCST operon did. When the umuDCST operon was specifically deleted from the chromosome of S. typhimurium TA2659, the resulting strain was not UV mutable and was more sensitive to the killing effect of UV irradiation than the parent strain was. Curing of the 60-MDa cryptic plasmid carrying the samAB operon did not influence the UV mutability of strain TA2659 but did increase its resistance to UV killing. A pSC101-derived plasmid carrying the samAB operon did not restore UV mutability to a umuD44 strain of E. coli, whereas pBR322- or pBluescript-derived plasmids carrying the samAB operon efficiently did restore UV mutability. We concluded that the umuDCST operon plays a major role in UV mutagenesis in S. typhimurium and that the ability of the samAB operon to promote UV mutagenesis is strongly affected by gene dosage. Possible reasons for the poor ability of samAB to promote UV mutagenesis when it is present on low-copy-number plasmids are discussed. Images PMID:1400244

  16. Adaptation effects in static postural control by providing simultaneous visual feedback of center of pressure and center of gravity.

    PubMed

    Takeda, Kenta; Mani, Hiroki; Hasegawa, Naoya; Sato, Yuki; Tanaka, Shintaro; Maejima, Hiroshi; Asaka, Tadayoshi

    2017-07-19

    The benefit of visual feedback of the center of pressure (COP) on quiet standing is still debatable. This study aimed to investigate the adaptation effects of visual feedback training using both the COP and center of gravity (COG) during quiet standing. Thirty-four healthy young adults were divided into three groups randomly (COP + COG, COP, and control groups). A force plate was used to calculate the coordinates of the COP in the anteroposterior (COP AP ) and mediolateral (COP ML ) directions. A motion analysis system was used to calculate the coordinates of the center of mass (COM) in both directions (COM AP and COM ML ). The coordinates of the COG in the AP direction (COG AP ) were obtained from the force plate signals. Augmented visual feedback was presented on a screen in the form of fluctuation circles in the vertical direction that moved upward as the COP AP and/or COG AP moved forward and vice versa. The COP + COG group received the real-time COP AP and COG AP feedback simultaneously, whereas the COP group received the real-time COP AP feedback only. The control group received no visual feedback. In the training session, the COP + COG group was required to maintain an even distance between the COP AP and COG AP and reduce the COG AP fluctuation, whereas the COP group was required to reduce the COP AP fluctuation while standing on a foam pad. In test sessions, participants were instructed to keep their standing posture as quiet as possible on the foam pad before (pre-session) and after (post-session) the training sessions. In the post-session, the velocity and root mean square of COM AP in the COP + COG group were lower than those in the control group. In addition, the absolute value of the sum of the COP - COM distances in the COP + COG group was lower than that in the COP group. Furthermore, positive correlations were found between the COM AP velocity and COP - COM parameters. The results suggest that the novel visual feedback training that incorporates the COP AP -COG AP interaction reduces postural sway better than the training using the COP AP alone during quiet standing. That is, even COP AP fluctuation around the COG AP would be effective in reducing the COM AP velocity.

  17. Induction of the Nitrate Assimilation nirA Operon and Protein-Protein Interactions in the Maturation of Nitrate and Nitrite Reductases in the Cyanobacterium Anabaena sp. Strain PCC 7120.

    PubMed

    Frías, José E; Flores, Enrique

    2015-07-01

    Nitrate is widely used as a nitrogen source by cyanobacteria, in which the nitrate assimilation structural genes frequently constitute the so-called nirA operon. This operon contains the genes encoding nitrite reductase (nirA), a nitrate/nitrite transporter (frequently an ABC-type transporter; nrtABCD), and nitrate reductase (narB). In the model filamentous cyanobacterium Anabaena sp. strain PCC 7120, which can fix N2 in specialized cells termed heterocysts, the nirA operon is expressed at high levels only in media containing nitrate or nitrite and lacking ammonium, a preferred nitrogen source. Here we examined the genes downstream of the nirA operon in Anabaena and found that a small open reading frame of unknown function, alr0613, can be cotranscribed with the operon. The next gene in the genome, alr0614 (narM), showed an expression pattern similar to that of the nirA operon, implying correlated expression of narM and the operon. A mutant of narM with an insertion mutation failed to produce nitrate reductase activity, consistent with the idea that NarM is required for the maturation of NarB. Both narM and narB mutants were impaired in the nitrate-dependent induction of the nirA operon, suggesting that nitrite is an inducer of the operon in Anabaena. It has previously been shown that the nitrite reductase protein NirA requires NirB, a protein likely involved in protein-protein interactions, to attain maximum activity. Bacterial two-hybrid analysis confirmed possible NirA-NirB and NarB-NarM interactions, suggesting that the development of both nitrite reductase and nitrate reductase activities in cyanobacteria involves physical interaction of the corresponding enzymes with their cognate partners, NirB and NarM, respectively. Nitrate is an important source of nitrogen for many microorganisms that is utilized through the nitrate assimilation system, which includes nitrate/nitrite membrane transporters and the nitrate and nitrite reductases. Many cyanobacteria assimilate nitrate, but regulation of the nitrate assimilation system varies in different cyanobacterial groups. In the N2-fixing, heterocyst-forming cyanobacteria, the nirA operon, which includes the structural genes for the nitrate assimilation system, is expressed in the presence of nitrate or nitrite if ammonium is not available to the cells. Here we studied the genes required for production of an active nitrate reductase, providing information on the nitrate-dependent induction of the operon, and found evidence for possible protein-protein interactions in the maturation of nitrate reductase and nitrite reductase. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  18. A predictive biophysical model of translational coupling to coordinate and control protein expression in bacterial operons

    PubMed Central

    Tian, Tian; Salis, Howard M.

    2015-01-01

    Natural and engineered genetic systems require the coordinated expression of proteins. In bacteria, translational coupling provides a genetically encoded mechanism to control expression level ratios within multi-cistronic operons. We have developed a sequence-to-function biophysical model of translational coupling to predict expression level ratios in natural operons and to design synthetic operons with desired expression level ratios. To quantitatively measure ribosome re-initiation rates, we designed and characterized 22 bi-cistronic operon variants with systematically modified intergenic distances and upstream translation rates. We then derived a thermodynamic free energy model to calculate de novo initiation rates as a result of ribosome-assisted unfolding of intergenic RNA structures. The complete biophysical model has only five free parameters, but was able to accurately predict downstream translation rates for 120 synthetic bi-cistronic and tri-cistronic operons with rationally designed intergenic regions and systematically increased upstream translation rates. The biophysical model also accurately predicted the translation rates of the nine protein atp operon, compared to ribosome profiling measurements. Altogether, the biophysical model quantitatively predicts how translational coupling controls protein expression levels in synthetic and natural bacterial operons, providing a deeper understanding of an important post-transcriptional regulatory mechanism and offering the ability to rationally engineer operons with desired behaviors. PMID:26117546

  19. 30 CFR 285.811 - When must I follow my Safety Management System?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... Activities Conducted Under SAPs, COPs and GAPs Safety Management Systems § 285.811 When must I follow my... activities described in your approved COP, SAP, or GAP. You must conduct all activities described in your approved COP, SAP, or GAP in accordance with the Safety Management System you described, as required by...

  20. Columbus Outdoor Pursuits: A Model Structure for Volunteer Based Outdoor Pursuits Programs.

    ERIC Educational Resources Information Center

    Gerckens, Ann E.

    Columbus Outdoor Pursuits (COP) is a volunteer-based nonprofit organization in Ohio that has offered outdoor educational and recreational opportunities to its members for 63 years. COP day and trip programs focus on outdoor activities such as bicycling, kayaking, hiking, and rock climbing. In addition, COP offers classes in the skills required for…

  1. 30 CFR 285.800 - How must I conduct my activities to comply with safety and environmental requirements?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ..., Inspections, and Facility Assessments for Activities Conducted Under SAPs, COPs and GAPs § 285.800 How must I... approved SAP, COP, or GAP, as required under §§ 285.615(c), 285.633(b), or 285.653(c). ...

  2. DHU1 negatively regulates UV-B signaling via its direct interaction with COP1 and RUP1.

    PubMed

    Kim, Sang-Hoon; Kim, Hani; Chung, Sunglan; Lee, Jae-Hoon

    2017-09-16

    Although DWD HYPERSENSITIVE TO UV-B 1 (DHU1) is reported to be a negative regulator in UV-B mediated cellular responses, its detailed role in UV-B signaling is still elusive. To further understand the action mechanism of DHU1 in UV-B response, physical and genetic interactions of DHU1 with various UV-B signaling components were investigated. Yeast two hybrid assay results suggested that DHU1 directly interacts with COP1 and RUP1, implying a functional connection with both COP1 and RUP1. In spite of the physical association between DHU1 and COP1, loss of DHU1 did not affect protein stability of COP1. Epistatic analysis showed that the functional loss of both DHU1 and UVR8 leads to alleviation of UV-B hypersensitivity displayed in dhu1-1. Moreover, phenotypic studies with dhu1-1 cop1-6 and dhu1-1 hy5-215 revealed that COP1 and HY5 are epistatic to DHU1, indicating that UV-B hypersensitivity of dhu1-1 requires both COP1 and HY5. In the case of dhu1-1 rup1-1, UV-B responsiveness was similar to that of both dhu1-1 and rup1-1, implying that DHU1 and RUP1 are required for each other's function. Collectively, these results show that the role of DHU1 as a negative regulator in UV-B response may be derived from its direct interaction with COP1 by sequestering COP1 from the active UVR8-COP1 complex, resulting in a decrease in the COP1 population that positively participates in UV-B signaling together with UVR8. Furthermore, this inhibitory role of DHU1 in UV-B signaling is likely to be functionally connected to RUP1. This study will serve as a platform to further understand more detailed action mechanism of DHU1 in UV-B response and DHU1-mediated core UV-B signaling in Arabidopsis. Copyright © 2017 Elsevier Inc. All rights reserved.

  3. 30 CFR 585.618 - What must I do upon completion of approved site assessment activities?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... assessment term, you timely submit a COP meeting the requirements of this subpart, or a complete FERC license... facilities in place on your lease during the time that BOEM reviews your COP for approval or FERC reviews... for facilities that are authorized to remain in place under your approved COP or approved FERC license...

  4. 30 CFR 585.618 - What must I do upon completion of approved site assessment activities?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... assessment term, you timely submit a COP meeting the requirements of this subpart, or a complete FERC license... facilities in place on your lease during the time that BOEM reviews your COP for approval or FERC reviews... for facilities that are authorized to remain in place under your approved COP or approved FERC license...

  5. 30 CFR 585.618 - What must I do upon completion of approved site assessment activities?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... assessment term, you timely submit a COP meeting the requirements of this subpart, or a complete FERC license... facilities in place on your lease during the time that BOEM reviews your COP for approval or FERC reviews... for facilities that are authorized to remain in place under your approved COP or approved FERC license...

  6. Identification of an operon, Pil-Chp, that controls twitching motility and virulence in Xylella fastidiosa.

    PubMed

    Cursino, Luciana; Galvani, Cheryl D; Athinuwat, Dusit; Zaini, Paulo A; Li, Yaxin; De La Fuente, Leonardo; Hoch, Harvey C; Burr, Thomas J; Mowery, Patricia

    2011-10-01

    Xylella fastidiosa is an important phytopathogenic bacterium that causes many serious plant diseases, including Pierce's disease of grapevines. Disease manifestation by X. fastidiosa is associated with the expression of several factors, including the type IV pili that are required for twitching motility. We provide evidence that an operon, named Pil-Chp, with genes homologous to those found in chemotaxis systems, regulates twitching motility. Transposon insertion into the pilL gene of the operon resulted in loss of twitching motility (pilL is homologous to cheA genes encoding kinases). The X. fastidiosa mutant maintained the type IV pili, indicating that the disrupted pilL or downstream operon genes are involved in pili function, and not biogenesis. The mutated X. fastidiosa produced less biofilm than wild-type cells, indicating that the operon contributes to biofilm formation. Finally, in planta the mutant produced delayed and less severe disease, indicating that the Pil-Chp operon contributes to the virulence of X. fastidiosa, presumably through its role in twitching motility.

  7. Direct colloid osmometry in healthy New World camelids.

    PubMed

    Quesada, Rolando J; Gorman, Maria Elena; Cebra, Christopher K; Verdugo, Claudio; Mosley, Craig A

    2011-06-01

    Direct colloid osmometry provides an objective assessment of the oncotic effects of crystalloid or colloidal fluid therapy, which is especially useful in monitoring fluid therapy of critically ill camelids due to their tendency toward nonspecific hypoproteinemia with increased risk of developing edema and ascites. The aims of this study were to measure colloid osmotic pressure (COP) of alpacas and llamas, determine its correlation with concentrations of total protein (TP) and total solids (TS), as well as both albumin (A) and globulin (G) concentrations in the same model (A+G), and evaluate the effects of sample type and storage conditions on COP. Blood was collected from clinically healthy alpacas (n=23) and llamas (n=22) into heparin tubes. COP of fresh whole blood (COP(FB) ) and plasma (COP(FP) ) was determined using a membrane osmometer. For 20 alpacas, COP of refrigerated whole blood (COP(RB) ) and frozen plasma (COP(FrP) ) was also measured. Correlations between COP(FB) and TS, TP, and A+G concentrations were assessed by simple and multiple regression analysis to model potential predictors. Median COP(FB) from alpacas (24.6 mmHg, range 19.3-28.1) was not significantly different from that of llamas (25.3 mmHg, range 22.5-33.7). Sample type or storage conditions did not affect COP. Measured COP had a strong positive linear correlation with TS, TP, and A+G concentrations in alpacas (r(2) =.7, .74, and .88, respectively). In llamas, COP correlated best with TS concentration (r(2) =.59), whereas correlation with TP and A+G concentrations was poor (r(2) =.19 and .25, respectively). COP can be measured using heparinized whole blood or plasma, either fresh or stored. Direct measurement is recommended whenever quantitative knowledge of COP is required in clinical or research setting. Further studies are needed to verify if the poor association of COP with TP found in this study can be generalized to llamas. ©2011 American Society for Veterinary Clinical Pathology.

  8. NATO initial common operational picture capability project

    NASA Astrophysics Data System (ADS)

    Fanti, Laura; Beach, David

    2002-08-01

    The Common Operational Picture (COP) capability can be defined as the ability to display on a single screen integrated views of the Recognized Maritime, Air and Ground Pictures, enriched by other tactical data, such as theater plans, assets, intelligence and logistics information. The purpose of the COP capability is to provide military forces a comprehensive view of the battle space, thereby enhancing situational awareness and the decision-making process across the military command and control spectrum. The availability of a COP capability throughout the command structure is a high priority operational requirement in NATO. A COP capability for NATO is being procured and implemented in an incremental way within the NATO Automated Information System (Bi-SC AIS) Functional Services programme under the coordination of the NATO Consultation, Command and Control Agency (NC3A) Integrated Programme Team 5 (IPT5). The NATO Initial COP (iCOP) capability project, first step of this evolutionary procurement, will provide an initial COP capability to NATO in a highly pragmatic and low-risk fashion, by using existing operational communications infrastructure and NATO systems, i.e. the NATO-Wide Integrated Command and Control Software for Air Operations (ICC), the Maritime Command and Control Information System (MCCIS), and the Joint Operations and Intelligence Information System (JOIIS), which will provide respectively the Recognized Air, Maritime and Ground Pictures. This paper gives an overview of the NATO Initial COP capability project, including its evolutionary implementation approach, and describes the technical solution selected to satisfy the urgent operational requirement in a timely and cost effective manner.

  9. Incorporation of a horizontally transferred gene into an operon during cnidarian evolution.

    PubMed

    Dana, Catherine E; Glauber, Kristine M; Chan, Titus A; Bridge, Diane M; Steele, Robert E

    2012-01-01

    Genome sequencing has revealed examples of horizontally transferred genes, but we still know little about how such genes are incorporated into their host genomes. We have previously reported the identification of a gene (flp) that appears to have entered the Hydra genome through horizontal transfer. Here we provide additional evidence in support of our original hypothesis that the transfer was from a unicellular organism, and we show that the transfer occurred in an ancestor of two medusozoan cnidarian species. In addition we show that the gene is part of a bicistronic operon in the Hydra genome. These findings identify a new animal phylum in which trans-spliced leader addition has led to the formation of operons, and define the requirements for evolution of an operon in Hydra. The identification of operons in Hydra also provides a tool that can be exploited in the construction of transgenic Hydra strains.

  10. c-Type Cytochrome Assembly Is a Key Target of Copper Toxicity within the Bacterial Periplasm

    PubMed Central

    Durand, Anne; Azzouzi, Asma; Bourbon, Marie-Line; Steunou, Anne-Soisig; Liotenberg, Sylviane; Maeshima, Akinori; Astier, Chantal; Argentini, Manuela; Saito, Shingo

    2015-01-01

    ABSTRACT In the absence of a tight control of copper entrance into cells, bacteria have evolved different systems to control copper concentration within the cytoplasm and the periplasm. Central to these systems, the Cu+ ATPase CopA plays a major role in copper tolerance and translocates copper from the cytoplasm to the periplasm. The fate of copper in the periplasm varies among species. Copper can be sequestered, oxidized, or released outside the cells. Here we describe the identification of CopI, a periplasmic protein present in many proteobacteria, and show its requirement for copper tolerance in Rubrivivax gelatinosus. The ΔcopI mutant is more susceptible to copper than the Cu+ ATPase copA mutant. CopI is induced by copper, localized in the periplasm and could bind copper. Interestingly, copper affects cytochrome c membrane complexes (cbb3 oxidase and photosystem) in both ΔcopI and copA-null mutants, but the causes are different. In the copA mutant, heme and chlorophyll synthesis are affected, whereas in ΔcopI mutant, the decrease is a consequence of impaired cytochrome c assembly. This impact on c-type cytochromes would contribute also to the copper toxicity in the periplasm of the wild-type cells when they are exposed to high copper concentrations. PMID:26396241

  11. Evaluation of the Role of the opgGH Operon in Yersinia pseudotuberculosis and Its Deletion during the Emergence of Yersinia pestis

    PubMed Central

    Quintard, Kévin; Dewitte, Amélie; Reboul, Angéline; Madec, Edwige; Bontemps-Gallo, Sébastien; Dondeyne, Jacqueline; Marceau, Michaël; Simonet, Michel

    2015-01-01

    The opgGH operon encodes glucosyltransferases that synthesize osmoregulated periplasmic glucans (OPGs) from UDP-glucose, using acyl carrier protein (ACP) as a cofactor. OPGs are required for motility, biofilm formation, and virulence in various bacteria. OpgH also sequesters FtsZ in order to regulate cell size according to nutrient availability. Yersinia pestis (the agent of flea-borne plague) lost the opgGH operon during its emergence from the enteropathogen Yersinia pseudotuberculosis. When expressed in OPG-negative strains of Escherichia coli and Dickeya dadantii, opgGH from Y. pseudotuberculosis restored OPGs synthesis, motility, and virulence. However, Y. pseudotuberculosis did not produce OPGs (i) under various growth conditions or (ii) when overexpressing its opgGH operon, its galUF operon (governing UDP-glucose), or the opgGH operon or Acp from E. coli. A ΔopgGH Y. pseudotuberculosis strain showed normal motility, biofilm formation, resistance to polymyxin and macrophages, and virulence but was smaller. Consistently, Y. pestis was smaller than Y. pseudotuberculosis when cultured at ≥37°C, except when the plague bacillus expressed opgGH. Y. pestis expressing opgGH grew normally in serum and within macrophages and was fully virulent in mice, suggesting that small cell size was not advantageous in the mammalian host. Lastly, Y. pestis expressing opgGH was able to infect Xenopsylla cheopis fleas normally. Our results suggest an evolutionary scenario whereby an ancestral Yersinia strain lost a factor required for OPG biosynthesis but kept opgGH (to regulate cell size). The opgGH operon was presumably then lost because OpgH-dependent cell size control became unnecessary. PMID:26150539

  12. 30 CFR 285.626 - What must I include in my COP?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... ENERGY ALTERNATE USES OF EXISTING FACILITIES ON THE OUTER CONTINENTAL SHELF Plans and Information Requirements Contents of the Construction and Operations Plan § 285.626 What must I include in my COP? (a) You...

  13. 30 CFR 285.622 - How do I submit my COP?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... ALTERNATE USES OF EXISTING FACILITIES ON THE OUTER CONTINENTAL SHELF Plans and Information Requirements Construction and Operations Plan for Commercial Leases § 285.622 How do I submit my COP? (a) You must submit...

  14. 30 CFR 285.621 - What must I demonstrate in my COP?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... RENEWABLE ENERGY ALTERNATE USES OF EXISTING FACILITIES ON THE OUTER CONTINENTAL SHELF Plans and Information Requirements Construction and Operations Plan for Commercial Leases § 285.621 What must I demonstrate in my COP...

  15. Ancient Origin of the Tryptophan Operon and the Dynamics of Evolutionary Change†

    PubMed Central

    Xie, Gary; Keyhani, Nemat O.; Bonner; Jensen, Roy A.

    2003-01-01

    The seven conserved enzymatic domains required for tryptophan (Trp) biosynthesis are encoded in seven genetic regions that are organized differently (whole-pathway operons, multiple partial-pathway operons, and dispersed genes) in prokaryotes. A comparative bioinformatics evaluation of the conservation and organization of the genes of Trp biosynthesis in prokaryotic operons should serve as an excellent model for assessing the feasibility of predicting the evolutionary histories of genes and operons associated with other biochemical pathways. These comparisons should provide a better understanding of possible explanations for differences in operon organization in different organisms at a genomics level. These analyses may also permit identification of some of the prevailing forces that dictated specific gene rearrangements during the course of evolution. Operons concerned with Trp biosynthesis in prokaryotes have been in a dynamic state of flux. Analysis of closely related organisms among the Bacteria at various phylogenetic nodes reveals many examples of operon scission, gene dispersal, gene fusion, gene scrambling, and gene loss from which the direction of evolutionary events can be deduced. Two milestone evolutionary events have been mapped to the 16S rRNA tree of Bacteria, one splitting the operon in two, and the other rejoining it by gene fusion. The Archaea, though less resolved due to a lesser genome representation, appear to exhibit more gene scrambling than the Bacteria. The trp operon appears to have been an ancient innovation; it was already present in the common ancestor of Bacteria and Archaea. Although the operon has been subjected, even in recent times, to dynamic changes in gene rearrangement, the ancestral gene order can be deduced with confidence. The evolutionary history of the genes of the pathway is discernible in rough outline as a vertical line of descent, with events of lateral gene transfer or paralogy enriching the analysis as interesting features that can be distinguished. As additional genomes are thoroughly analyzed, an increasingly refined resolution of the sequential evolutionary steps is clearly possible. These comparisons suggest that present-day trp operons that possess finely tuned regulatory features are under strong positive selection and are able to resist the disruptive evolutionary events that may be experienced by simpler, poorly regulated operons. PMID:12966138

  16. Escherichia coli mutant with altered respiratory control of the frd operon.

    PubMed Central

    Iuchi, S; Kuritzkes, D R; Lin, E C

    1985-01-01

    In wild-type Escherichia coli, fumarate reductase encoded by the frd operon is inducible by its substrate in the absence of molecular oxygen and nitrate. Synthesis of this enzyme under permissive conditions requires the fnr+ gene product, which is believed to be a pleiotropic regulatory protein that activates transcription. A spontaneous mutant was isolated in which the expression of the frd operon no longer depended on the presence of fumarate or the fnr+ gene product. Aerobic repression of the operon was abolished, but nitrate repression remained intact. Transductional analysis showed that the mutation was closely linked to the frd locus. The mutant phenotype strongly suggests that repression by molecular oxygen and nitrate is mediated by different mechanisms. PMID:3882660

  17. 30 CFR 285.634 - What activities require a revision to my COP, and when will MMS approve the revision?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...: (1) Designed not to cause undue harm or damage to natural resources; life (including human and... 30 Mineral Resources 2 2010-07-01 2010-07-01 false What activities require a revision to my COP, and when will MMS approve the revision? 285.634 Section 285.634 Mineral Resources MINERALS MANAGEMENT...

  18. 30 CFR 285.815 - What must I do if I have facility damage or an equipment failure?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... Environmental and Safety Management, Inspections, and Facility Assessments for Activities Conducted Under SAPs..., MMS may require you to revise your SAP, COP, or GAP to describe how you will address the facility damage or failure as required by § 285.634 (COP), § 285.617 (SAP), § 285.655 (GAP). You must submit a...

  19. Unprecedented high-resolution view of bacterial operon architecture revealed by RNA sequencing.

    PubMed

    Conway, Tyrrell; Creecy, James P; Maddox, Scott M; Grissom, Joe E; Conkle, Trevor L; Shadid, Tyler M; Teramoto, Jun; San Miguel, Phillip; Shimada, Tomohiro; Ishihama, Akira; Mori, Hirotada; Wanner, Barry L

    2014-07-08

    We analyzed the transcriptome of Escherichia coli K-12 by strand-specific RNA sequencing at single-nucleotide resolution during steady-state (logarithmic-phase) growth and upon entry into stationary phase in glucose minimal medium. To generate high-resolution transcriptome maps, we developed an organizational schema which showed that in practice only three features are required to define operon architecture: the promoter, terminator, and deep RNA sequence read coverage. We precisely annotated 2,122 promoters and 1,774 terminators, defining 1,510 operons with an average of 1.98 genes per operon. Our analyses revealed an unprecedented view of E. coli operon architecture. A large proportion (36%) of operons are complex with internal promoters or terminators that generate multiple transcription units. For 43% of operons, we observed differential expression of polycistronic genes, despite being in the same operons, indicating that E. coli operon architecture allows fine-tuning of gene expression. We found that 276 of 370 convergent operons terminate inefficiently, generating complementary 3' transcript ends which overlap on average by 286 nucleotides, and 136 of 388 divergent operons have promoters arranged such that their 5' ends overlap on average by 168 nucleotides. We found 89 antisense transcripts of 397-nucleotide average length, 7 unannotated transcripts within intergenic regions, and 18 sense transcripts that completely overlap operons on the opposite strand. Of 519 overlapping transcripts, 75% correspond to sequences that are highly conserved in E. coli (>50 genomes). Our data extend recent studies showing unexpected transcriptome complexity in several bacteria and suggest that antisense RNA regulation is widespread. Importance: We precisely mapped the 5' and 3' ends of RNA transcripts across the E. coli K-12 genome by using a single-nucleotide analytical approach. Our resulting high-resolution transcriptome maps show that ca. one-third of E. coli operons are complex, with internal promoters and terminators generating multiple transcription units and allowing differential gene expression within these operons. We discovered extensive antisense transcription that results from more than 500 operons, which fully overlap or extensively overlap adjacent divergent or convergent operons. The genomic regions corresponding to these antisense transcripts are highly conserved in E. coli (including Shigella species), although it remains to be proven whether or not they are functional. Our observations of features unearthed by single-nucleotide transcriptome mapping suggest that deeper layers of transcriptional regulation in bacteria are likely to be revealed in the future. Copyright © 2014 Conway et al.

  20. 30 CFR 285.659 - What requirements must I include in my SAP, COP, or GAP regarding air quality?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ..., or GAP regarding air quality? 285.659 Section 285.659 Mineral Resources BUREAU OF OCEAN ENERGY... Pipeline Deviations § 285.659 What requirements must I include in my SAP, COP, or GAP regarding air quality..., according to the following table. ER29AP09.130 (b) For air quality modeling that you perform in support of...

  1. 30 CFR 585.800 - How must I conduct my activities to comply with safety and environmental requirements?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... for Activities Conducted Under SAPs, COPs and GAPs § 585.800 How must I conduct my activities to.... (b) You must certify compliance with those terms and conditions identified in your approved SAP, COP...

  2. 30 CFR 585.800 - How must I conduct my activities to comply with safety and environmental requirements?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... for Activities Conducted Under SAPs, COPs and GAPs § 585.800 How must I conduct my activities to.... (b) You must certify compliance with those terms and conditions identified in your approved SAP, COP...

  3. 30 CFR 585.800 - How must I conduct my activities to comply with safety and environmental requirements?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... for Activities Conducted Under SAPs, COPs and GAPs § 585.800 How must I conduct my activities to.... (b) You must certify compliance with those terms and conditions identified in your approved SAP, COP...

  4. 30 CFR 285.416 - How do I request a lease or grant suspension?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... SAP, COP, or GAP, as required, you may request a suspension to extend the preliminary or site assessment term of your lease or grant that includes a revised schedule for submission of a SAP, COP, or GAP...

  5. α2-COP is involved in early secretory traffic in Arabidopsis and is required for plant growth

    PubMed Central

    Gimeno-Ferrer, Fátima; Pastor-Cantizano, Noelia; Bernat-Silvestre, César; Selvi-Martínez, Pilar; Vera-Sirera, Francisco; Gao, Caiji; Perez-Amador, Miguel Angel; Jiang, Liwen; Aniento, Fernando

    2017-01-01

    Abstract COP (coat protein) I-coated vesicles mediate intra-Golgi transport and retrograde transport from the Golgi to the endoplasmic reticulum. These vesicles form through the action of the small GTPase ADP-ribosylation factor 1 (ARF1) and the COPI heptameric protein complex (coatomer), which consists of seven subunits (α-, β-, β′-, γ-, δ-, ε- and ζ-COP). In contrast to mammals and yeast, several isoforms for coatomer subunits, with the exception of γ and δ, have been identified in Arabidopsis. To understand the role of COPI proteins in plant biology, we have identified and characterized a loss-of-function mutant of α2-COP, an Arabidopsis α-COP isoform. The α2-cop mutant displayed defects in plant growth, including small rosettes, stems and roots and mislocalization of p24δ5, a protein of the p24 family containing a C-terminal dilysine motif involved in COPI binding. The α2-cop mutant also exhibited abnormal morphology of the Golgi apparatus. Global expression analysis of the α2-cop mutant revealed altered expression of plant cell wall-associated genes. In addition, a strong upregulation of SEC31A, which encodes a subunit of the COPII coat, was observed in the α2-cop mutant; this also occurs in a mutant of a gene upstream of COPI assembly, GNL1, which encodes an ARF-guanine nucleotide exchange factor (GEF). These findings suggest that loss of α2-COP affects the expression of secretory pathway genes. PMID:28025315

  6. Experimental heat and mass transfer of the separated and coupled rotating desiccant wheel and heat wheel

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

    Enteria, Napoleon; Yoshino, Hiroshi; Mochida, Akashi

    The experimental evaluation of the separated and coupled rotating desiccant wheel and heat wheel is reported. The study aims to investigate the performance of the desiccant wheel and of the heat wheel both when operated separately and jointly. The performance evaluation of the desiccant wheel is based on its moisture removal capacity (MRC), moisture removal regeneration (MRR), and moisture mass balance (MMB). In addition, the study used the total energy balance (TEB), sensible coefficient of performance (COP{sub Sensible}), latent coefficient of performance (COP{sub Latent}) and, total coefficient of performance (COP{sub Total}). The performance of the heat wheel is based onmore » its effectiveness. The COP{sub Sensible}, COP{sub Latent} and, COP{sub Total} are used in the performance evaluation of the coupled desiccant wheel and heat wheel. The general results of the study show that the MRC, MRR and MMB coupled with the TEB, COP{sub Latent}, COP{sub Sensible} and COP{sub Total} predict adequately the performance of the desiccant wheel. In addition, the coupled operation of the desiccant wheel and heat wheel, contributed to the reduction of the external thermal energy requirement for the regeneration of the desiccant wheel. This study can be applied in other researches seeking evaluation of the desiccant wheel, heat wheel, and their combined operation. Moreover, the data presented here are significant for the desiccant wheel benchmarking and for evaluation of the desiccant wheel models. (author)« less

  7. 30 CFR 585.416 - How do I request a lease or grant suspension?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... submit a SAP, COP, or GAP, as required, you may request a suspension to extend the preliminary or site assessment term of your lease or grant that includes a revised schedule for submission of a SAP, COP, or GAP...

  8. 30 CFR 585.416 - How do I request a lease or grant suspension?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... submit a SAP, COP, or GAP, as required, you may request a suspension to extend the preliminary or site assessment term of your lease or grant that includes a revised schedule for submission of a SAP, COP, or GAP...

  9. 30 CFR 285.810 - What must I include in my Safety Management System?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... Activities Conducted Under SAPs, COPs and GAPs Safety Management Systems § 285.810 What must I include in my... your COP (provided under § 285.627(d)) and, when required by this part, your SAP (as provided in § 285...

  10. 30 CFR 585.416 - How do I request a lease or grant suspension?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... submit a SAP, COP, or GAP, as required, you may request a suspension to extend the preliminary or site assessment term of your lease or grant that includes a revised schedule for submission of a SAP, COP, or GAP...

  11. Supporting Emerging Disciplines with e-Communities: Needs and Benefits

    PubMed Central

    Butler, Brian S; Schleyer, Titus K; Weiss, Patricia M; Wang, Xiaoqing; Thyvalikakath, Thankam P; Hatala, Courtney L; Naderi, Reza A

    2008-01-01

    Background Science has developed from a solitary pursuit into a team-based collaborative activity and, more recently, into a multidisciplinary research enterprise. The increasingly collaborative character of science, mandated by complex research questions and problems that require many competencies, requires that researchers lower the barriers to the creation of collaborative networks of experts, such as communities of practice (CoPs). Objectives The aim was to assess the information needs of prospective members of a CoP in an emerging field, dental informatics, and to evaluate their expectations of an e-community in order to design a suitable electronic infrastructure. Methods A Web-based survey instrument was designed and administered to 2768 members of the target audience. Benefit expectations were analyzed for their relationship to (1) the respondents’ willingness to participate in the CoP and (2) their involvement in funded research. Two raters coded the respondents’ answers regarding expected benefits using a 14-category coding scheme (Kappa = 0.834). Results The 256 respondents (11.1% response rate) preferred electronic resources over traditional print material to satisfy their information needs. The most frequently expected benefits from participation in the CoP were general information (85% of respondents), peer networking (31.1%), and identification of potential collaborators and/or research opportunities (23.2%). Conclusions The competitive social-information environment in which CoPs are embedded presents both threats to sustainability and opportunities for greater integration and impact. CoP planners seeking to support the development of emerging biomedical science disciplines should blend information resources, social search and filtering, and visibility mechanisms to provide a portfolio of social and information benefits. Assessing benefit expectations and alternatives provides useful information for CoP planners seeking to prioritize community infrastructure development and encourage participation. PMID:18653443

  12. Crosstalk between virulence loci: regulation of Salmonella enterica pathogenicity island 1 (SPI-1) by products of the std fimbrial operon.

    PubMed

    López-Garrido, Javier; Casadesús, Josep

    2012-01-01

    Invasion of intestinal epithelial cells is a critical step in Salmonella infection and requires the expression of genes located in Salmonella pathogenicity island 1 (SPI-1). A key factor for SPI-1 expression is DNA adenine (Dam) methylation, which activates synthesis of the SPI-1 transcriptional activator HilD. Dam-dependent regulation of hilD is postranscriptional (and therefore indirect), indicating the involvement of unknown cell functions under Dam methylation control. A genetic screen has identified the std fimbrial operon as the missing link between Dam methylation and SPI-1. We show that all genes in the std operon are part of a single transcriptional unit, and describe three previously uncharacterized ORFs (renamed stdD, stdE, and stdF). We present evidence that two such loci (stdE and stdF) are involved in Dam-dependent control of Salmonella SPI-1: in a Dam(-) background, deletion of stdE or stdF suppresses SPI-1 repression; in a Dam(+) background, constitutive expression of StdE and/or StdF represses SPI-1. Repression of SPI-1 by products of std operon explains the invasion defect of Salmonella Dam(-) mutants, which constitutively express the std operon. Dam-dependent repression of std in the ileum may be required to permit invasion, as indicated by two observations: constitutive expression of StdE and StdF reduces invasion of epithelial cells in vitro (1,000 fold) and attenuates Salmonella virulence in the mouse model (>60 fold). In turn, crosstalk between std and SPI-1 may play a role in intestinal infections by preventing expression of SPI-1 in the caecum, an intestinal compartment in which the std operon is known to be expressed.

  13. 30 CFR 285.628 - How will MMS process my COP?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 30 Mineral Resources 2 2011-07-01 2011-07-01 false How will MMS process my COP? 285.628 Section 285.628 Mineral Resources BUREAU OF OCEAN ENERGY MANAGEMENT, REGULATION, AND ENFORCEMENT, DEPARTMENT... CONTINENTAL SHELF Plans and Information Requirements Contents of the Construction and Operations Plan § 285...

  14. 78 FR 16830 - Submission for OMB Review; Comment Request

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-03-19

    ... submit to the Office of Management and Budget (OMB) for clearance the following proposal for collection... agreements for research supporting the management of coastal ecosystems. The statutory authority for COP is... project final report using COP formats. All of these requirements are needed for better evaluation of...

  15. Improved determination of dynamic balance using the centre of mass and centre of pressure inclination variables in a complete golf swing cycle.

    PubMed

    Choi, Ahnryul; Sim, Taeyong; Mun, Joung Hwan

    2016-01-01

    Golf requires proper dynamic balance to accurately control the club head through a harmonious coordination of each human segment and joint. In this study, we evaluated the ability for dynamic balance during a golf swing by using the centre of mass (COM)-centre of pressure (COP) inclination variables. Twelve professional, 13 amateur and 10 novice golfers participated in this study. Six infrared cameras, two force platforms and SB-Clinic software were used to measure the net COM and COP trajectories. In order to evaluate dynamic balance ability, the COM-COP inclination angle, COM-COP inclination angular velocity and normalised COM-COP inclination angular jerk were used. Professional golfer group revealed a smaller COM-COP inclination angle and angular velocity than novice golfer group in the lead/trail direction (P < 0.01). In the normalised COM-COP inclination angular jerk, the professional golfer group showed a lower value than the other two groups in all directions. Professional golfers tend to exhibit improved dynamic balance, and this can be attributed to the neuromusculoskeletal system that maintains balance with proper postural control. This study has the potential to allow for an evaluation of the dynamic balance mechanism and will provide useful basic information for swing training and prevention of golf injuries.

  16. Functional Analysis of the Magnetosome Island in Magnetospirillum gryphiswaldense: The mamAB Operon Is Sufficient for Magnetite Biomineralization

    PubMed Central

    Lohße, Anna; Ullrich, Susanne; Katzmann, Emanuel; Borg, Sarah; Wanner, Gerd; Richter, Michael; Voigt, Birgit; Schweder, Thomas; Schüler, Dirk

    2011-01-01

    Bacterial magnetosomes are membrane-enveloped, nanometer-sized crystals of magnetite, which serve for magnetotactic navigation. All genes implicated in the synthesis of these organelles are located in a conserved genomic magnetosome island (MAI). We performed a comprehensive bioinformatic, proteomic and genetic analysis of the MAI in Magnetospirillum gryphiswaldense. By the construction of large deletion mutants we demonstrate that the entire region is dispensable for growth, and the majority of MAI genes have no detectable function in magnetosome formation and could be eliminated without any effect. Only <25% of the region comprising four major operons could be associated with magnetite biomineralization, which correlated with high expression of these genes and their conservation among magnetotactic bacteria. Whereas only deletion of the mamAB operon resulted in the complete loss of magnetic particles, deletion of the conserved mms6, mamGFDC, and mamXY operons led to severe defects in morphology, size and organization of magnetite crystals. However, strains in which these operons were eliminated together retained the ability to synthesize small irregular crystallites, and weakly aligned in magnetic fields. This demonstrates that whereas the mamGFDC, mms6 and mamXY operons have crucial and partially overlapping functions for the formation of functional magnetosomes, the mamAB operon is the only region of the MAI, which is necessary and sufficient for magnetite biomineralization. Our data further reduce the known minimal gene set required for magnetosome formation and will be useful for future genome engineering approaches. PMID:22043287

  17. Regulation of Lactobacillus casei Sorbitol Utilization Genes Requires DNA-Binding Transcriptional Activator GutR and the Conserved Protein GutM▿

    PubMed Central

    Alcántara, Cristina; Sarmiento-Rubiano, Luz Adriana; Monedero, Vicente; Deutscher, Josef; Pérez-Martínez, Gaspar; Yebra, María J.

    2008-01-01

    Sequence analysis of the five genes (gutRMCBA) downstream from the previously described sorbitol-6-phosphate dehydrogenase-encoding Lactobacillus casei gutF gene revealed that they constitute a sorbitol (glucitol) utilization operon. The gutRM genes encode putative regulators, while the gutCBA genes encode the EIIC, EIIBC, and EIIA proteins of a phosphoenolpyruvate-dependent sorbitol phosphotransferase system (PTSGut). The gut operon is transcribed as a polycistronic gutFRMCBA messenger, the expression of which is induced by sorbitol and repressed by glucose. gutR encodes a transcriptional regulator with two PTS-regulated domains, a galactitol-specific EIIB-like domain (EIIBGat domain) and a mannitol/fructose-specific EIIA-like domain (EIIAMtl domain). Its inactivation abolished gut operon transcription and sorbitol uptake, indicating that it acts as a transcriptional activator. In contrast, cells carrying a gutB mutation expressed the gut operon constitutively, but they failed to transport sorbitol, indicating that EIIBCGut negatively regulates GutR. A footprint analysis showed that GutR binds to a 35-bp sequence upstream from the gut promoter. A sequence comparison with the presumed promoter region of gut operons from various firmicutes revealed a GutR consensus motif that includes an inverted repeat. The regulation mechanism of the L. casei gut operon is therefore likely to be operative in other firmicutes. Finally, gutM codes for a conserved protein of unknown function present in all sequenced gut operons. A gutM mutant, the first constructed in a firmicute, showed drastically reduced gut operon expression and sorbitol uptake, indicating a regulatory role also for GutM. PMID:18676710

  18. Sulfate-Dependent Repression of Genes That Function in Organosulfur Metabolism in Bacillus subtilis Requires Spx

    PubMed Central

    Erwin, Kyle N.; Nakano, Shunji; Zuber, Peter

    2005-01-01

    Oxidative stress in Bacillus subtilis results in the accumulation of Spx protein, which exerts both positive and negative transcriptional control over a genome-wide scale through its interaction with the RNA polymerase α subunit. Previous microarray transcriptome studies uncovered a unique class of genes that are controlled by Spx-RNA polymerase interaction under normal growth conditions that do not promote Spx overproduction. These genes were repressed by Spx when sulfate was present as a sole sulfur source. The genes include those of the ytmI, yxeI, and ssu operons, which encode products resembling proteins that function in the uptake and desulfurization of organic sulfur compounds. Primer extension and analysis of operon-lacZ fusion expression revealed that the operons are repressed by sulfate and cysteine; however, Spx functioned only in sulfate-dependent repression. Both the ytmI operon and the divergently transcribed ytlI, encoding a LysR-type regulator that positively controls ytmI operon transcription, are repressed by Spx in sulfate-containing media. The CXXC motif of Spx, which is necessary for redox sensitive control of Spx activity in response to oxidative stress, is not required for sulfate-dependent repression. The yxeL-lacZ and ssu-lacZ fusions were also repressed in an Spx-dependent manner in media containing sulfate as the sole sulfur source. This work uncovers a new role for Spx in the control of sulfur metabolism in a gram-positive bacterium under nonstressful growth conditions. PMID:15937167

  19. 30 CFR 585.705 - When must I use a Certified Verification Agent (CVA)?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... with your SAP under § 585.610(a)(9), COP under § 585.626(b)(20), or GAP under § 585.645(c)(5). (1) BOEM... decision on your SAP, COP, or GAP. (2) If BOEM does not waive the requirement for a CVA, you may file an...

  20. 30 CFR 585.705 - When must I use a Certified Verification Agent (CVA)?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... with your SAP under § 585.610(a)(9), COP under § 585.626(b)(20), or GAP under § 585.645(c)(5). (1) BOEM... decision on your SAP, COP, or GAP. (2) If BOEM does not waive the requirement for a CVA, you may file an...

  1. 30 CFR 285.626 - What must I include in my COP?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 30 Mineral Resources 2 2011-07-01 2011-07-01 false What must I include in my COP? 285.626 Section 285.626 Mineral Resources BUREAU OF OCEAN ENERGY MANAGEMENT, REGULATION, AND ENFORCEMENT, DEPARTMENT... CONTINENTAL SHELF Plans and Information Requirements Contents of the Construction and Operations Plan § 285...

  2. Kinematics of center of mass and center of pressure predict friction requirement at shoe-floor interface during walking.

    PubMed

    Yamaguchi, Takeshi; Yano, Masaru; Onodera, Hiroshi; Hokkirigawa, Kazuo

    2013-06-01

    We aimed to determine whether inclination angles of the line connecting the whole body center of mass (COM) to the center of pressure (COP) (COM-COP angle) help predict the required coefficient of friction (RCOF) in young adult males during the weight acceptance and push-off phases in transient movements such as turning, gait termination and initiation, and steady-state movements such as straight walking. Seventeen healthy young adult males were asked to (1) walk in a straight line, (2) turn 60° with either foot (step and spin turns), and (3) initiate and terminate walking on a dry level floor. Peak absolute values of the ratio between resultant horizontal and vertical ground reaction forces during the weight acceptance and push-off phases (RCOFh and RCOFt, respectively) were calculated. COM-COP angles θh and θt at the instant of RCOFh and RCOFt, respectively, were also calculated. Bivariate regression analysis demonstrated that the |θh| and |θt| tangents were significant predictors of RCOFh (R = 0.878; R(2) = 0.770; p<0.001) and RCOFt (R = 0.918; R(2) = 0.843; p<0.001), respectively. The results suggest that COM and COP kinematics (i.e., the COM-COP angle) serve as a predictor of friction requirement during the weight acceptance and push-off phases in steady-state movements such as straight walking and transient movements such as turning as well as gait termination and initiation. Copyright © 2012 Elsevier B.V. All rights reserved.

  3. Pneumonia risk stratification in tropical Australia: does the SMART-COP score apply?

    PubMed

    Davis, Joshua S; Cross, Gail B; Charles, Patrick G P; Currie, Bart J; Anstey, Nicholas M; Cheng, Allen C

    2010-02-01

    To examine the performance in tropical northern Australia of SMART-COP, a simple scoring system developed in temperate Australia to predict the need for intensive respiratory or vasopressor support (IRVS) in pneumonia patients. A prospective observational study of patients admitted to Royal Darwin Hospital in the Northern Territory with sepsis between August 2007 and May 2008. Chest x-rays were reviewed to confirm pneumonia, and each patient's SMART-COP score was assessed against the need for IRVS. Of 206 patients presenting with radiologically confirmed pneumonia, 184 were eligible for inclusion. The mean age of patients was 50.1 years, 65% were Indigenous and 56% were men. Overall, 38 patients (21%) required IRVS, and 18 patients (10%) died by Day 30. A SMART-COP score of >or= 3 had a sensitivity of only 71% for predicting the need for IRVS and 67% for 30-day mortality. As the variables most strongly associated with IRVS were serum albumin level < 35 g/L (odds ratio, 6.8) and Indigenous status (odds ratio, 2.3), we tested a modified scoring system (SMART-COP) that used a higher weighting for albumin and included Indigenous status. A SMART-COP score of >or= 3 had a sensitivity of 97% for IRVS and 100% for 30-day mortality. The SMART-COP score underestimates the severity of pneumonia in tropical northern Australia, but can be improved by using locally relevant additions.

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

    PubMed

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

    2011-06-01

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

  5. ATES/heat pump simulations performed with ATESSS code

    NASA Astrophysics Data System (ADS)

    Vail, L. W.

    1989-01-01

    Modifications to the Aquifer Thermal Energy Storage System Simulator (ATESSS) allow simulation of aquifer thermal energy storage (ATES)/heat pump systems. The heat pump algorithm requires a coefficient of performance (COP) relationship of the form: COP = COP sub base + alpha (T sub ref minus T sub base). Initial applications of the modified ATES code to synthetic building load data for two sizes of buildings in two U.S. cities showed insignificant performance advantage of a series ATES heat pump system over a conventional groundwater heat pump system. The addition of algorithms for a cooling tower and solar array improved performance slightly. Small values of alpha in the COP relationship are the principal reason for the limited improvement in system performance. Future studies at Pacific Northwest Laboratory (PNL) are planned to investigate methods to increase system performance using alternative system configurations and operations scenarios.

  6. Myeloid leukemia factor 1 regulates p53 by suppressing COP1 via COP9 signalosome subunit 3.

    PubMed

    Yoneda-Kato, Noriko; Tomoda, Kiichiro; Umehara, Mari; Arata, Yukinobu; Kato, Jun-ya

    2005-05-04

    Myeloid leukemia factor 1 (MLF1) was first identified as the leukemic fusion protein NPM-MLF1 generated by the t(3;5)(q25.1;q34) chromosomal translocation. Although MLF1 expresses normally in a variety of tissues including hematopoietic stem cells and the overexpression of MLF1 correlates with malignant transformation in human cancer, little is known about how MLF1 is involved in the regulation of cell growth. Here we show that MLF1 is a negative regulator of cell cycle progression functioning upstream of the tumor suppressor p53. MLF1 induces p53-dependent cell cycle arrest in murine embryonic fibroblasts. This action requires a novel binding partner, subunit 3 of the COP9 signalosome (CSN3). A reduction in the level of CSN3 protein with small interfering RNA abrogated MLF1-induced G1 arrest and impaired the activation of p53 by genotoxic stress. Furthermore, ectopic MLF1 expression and CSN3 knockdown inversely affect the endogenous level of COP1, a ubiquitin ligase for p53. Exogenous expression of COP1 overcomes MLF1-induced growth arrest. These results indicate that MLF1 is a critical regulator of p53 and suggest its involvement in leukemogenesis through a novel CSN3-COP1 pathway.

  7. 30 CFR 285.659 - What requirements must I include in my SAP, COP, or GAP regarding air quality?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ..., or GAP regarding air quality? 285.659 Section 285.659 Mineral Resources MINERALS MANAGEMENT SERVICE... must I include in my SAP, COP, or GAP regarding air quality? (a) You must comply with the Clean Air Act...) For air quality modeling that you perform in support of the activities proposed in your plan, you...

  8. Extrachromosomal Nucleolus-Like Compartmentalization by a Plasmid-Borne Ribosomal RNA Operon and Its Role in Nucleoid Compaction.

    PubMed

    Mata Martin, Carmen; Sun, Zhe; Zhou, Yan Ning; Jin, Ding Jun

    2018-01-01

    In the fast-growing Escherichia coli cells, RNA polymerase (RNAP) molecules are concentrated and form foci at clusters of ribosomal RNA (rRNA) operons resembling eukaryotic nucleolus. The bacterial nucleolus-like organization, spatially compartmentalized at the surface of the compact bacterial chromosome (nucleoid), serves as transcription factories for rRNA synthesis and ribosome biogenesis, which influences the organization of the nucleoid. Unlike wild type that has seven rRNA operons in the genome in a mutant that has six (Δ6 rrn ) rRNA operons deleted in the genome, there are no apparent transcription foci and the nucleoid becomes uncompacted, indicating that formation of RNAP foci requires multiple copies of rRNA operons clustered in space and is critical for nucleoid compaction. It has not been determined, however, whether a multicopy plasmid-borne rRNA operon (p rrnB ) could substitute the multiple chromosomal rRNA operons for the organization of the bacterial nucleolus-like structure in the mutants of Δ6 rrn and Δ7 rrn that has all seven rRNA operons deleted in the genome. We hypothesized that extrachromosomal nucleolus-like structures are similarly organized and functional in trans from p rrnB in these mutants. In this report, using multicolor images of three-dimensional superresolution Structured Illumination Microscopy (3D-SIM), we determined the distributions of both RNAP and NusB that are a transcription factor involved in rRNA synthesis and ribosome biogenesis, p rrnB clustering, and nucleoid structure in these two mutants in response to environmental cues. Our results found that the extrachromosomal nucleolus-like organization tends to be spatially located at the poles of the mutant cells. In addition, formation of RNAP foci at the extrachromosomal nucleolus-like structure condenses the nucleoid, supporting the idea that active transcription at the nucleolus-like organization is a driving force in nucleoid compaction.

  9. Association with β-COP Regulates the Trafficking of the Newly Synthesized Na,K-ATPase*

    PubMed Central

    Morton, Michael J.; Farr, Glen A.; Hull, Michael; Capendeguy, Oihana; Horisberger, Jean-Daniel; Caplan, Michael J.

    2010-01-01

    Plasma membrane expression of the Na,K-ATPase requires assembly of its α- and β-subunits. Using a novel labeling technique to identify Na,K-ATPase partner proteins, we detected an interaction between the Na,K-ATPase α-subunit and the coat protein, β-COP, a component of the COP-I complex. When expressed in the absence of the Na,K-ATPase β-subunit, the Na,K-ATPase α-subunit interacts with β-COP, is retained in the endoplasmic reticulum, and is targeted for degradation. In the presence of the Na,K-ATPase β-subunit, the α-subunit does not interact with β-COP and traffics to the plasma membrane. Pulse-chase experiments demonstrate that in cells expressing both the Na,K-ATPase α- and β-subunits, newly synthesized α-subunit associates with β-COP immediately after its synthesis but that this interaction does not constitute an obligate intermediate in the assembly of the α- and β-subunits to form the pump holoenzyme. The interaction with β-COP was reduced by mutating a dibasic motif at Lys54 in the Na,K-ATPase α-subunit. This mutant α-subunit is not retained in the endoplasmic reticulum and reaches the plasma membrane, even in the absence of Na,K-ATPase β-subunit expression. Although the Lys54 α-subunit reaches the cell surface without need for β-subunit assembly, it is only functional as an ion-transporting ATPase in the presence of the β-subunit. PMID:20801885

  10. Silencing of Essential Genes within a Highly Coordinated Operon in Escherichia coli.

    PubMed

    Goh, Shan; Hohmeier, Angela; Stone, Timothy C; Offord, Victoria; Sarabia, Francisco; Garcia-Ruiz, Cristina; Good, Liam

    2015-08-15

    Essential bacterial genes located within operons are particularly challenging to study independently because of coordinated gene expression and the nonviability of knockout mutants. Essentiality scores for many operon genes remain uncertain. Antisense RNA (asRNA) silencing or in-frame gene disruption of genes may help establish essentiality but can lead to polar effects on genes downstream or upstream of the target gene. Here, the Escherichia coli ribF-ileS-lspA-fkpB-ispH operon was used to evaluate the possibility of independently studying an essential gene using expressed asRNA and target gene overexpression to deregulate coupled expression. The gene requirement for growth in conditional silencing strains was determined by the relationship of target mRNA reduction with growth inhibition as the minimum transcript level required for 50% growth (MTL50). Mupirocin and globomycin, the protein inhibitors of IleS and LspA, respectively, were used in sensitization assays of strains containing both asRNA-expressing and open reading frame-expressing plasmids to examine deregulation of the overlapping ileS-lspA genes. We found upstream and downstream polar silencing effects when either ileS or lspA was silenced, indicating coupled expression. Weighted MTL50 values (means and standard deviations) of ribF, ileS, and lspA were 0.65 ± 0.18, 0.64 ± 0.06, and 0.76 ± 0.10, respectively. However, they were not significantly different (P = 0.71 by weighted one-way analysis of variance). The gene requirement for ispH could not be determined due to insufficient growth reduction. Mupirocin and globomycin sensitization experiments indicated that ileS-lspA expression could not be decoupled. The results highlight the inherent challenges associated with genetic analyses of operons; however, coupling of essential genes may provide opportunities to improve RNA-silencing antimicrobials. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  11. Silencing of Essential Genes within a Highly Coordinated Operon in Escherichia coli

    PubMed Central

    Hohmeier, Angela; Stone, Timothy C.; Offord, Victoria; Sarabia, Francisco; Garcia-Ruiz, Cristina; Good, Liam

    2015-01-01

    Essential bacterial genes located within operons are particularly challenging to study independently because of coordinated gene expression and the nonviability of knockout mutants. Essentiality scores for many operon genes remain uncertain. Antisense RNA (asRNA) silencing or in-frame gene disruption of genes may help establish essentiality but can lead to polar effects on genes downstream or upstream of the target gene. Here, the Escherichia coli ribF-ileS-lspA-fkpB-ispH operon was used to evaluate the possibility of independently studying an essential gene using expressed asRNA and target gene overexpression to deregulate coupled expression. The gene requirement for growth in conditional silencing strains was determined by the relationship of target mRNA reduction with growth inhibition as the minimum transcript level required for 50% growth (MTL50). Mupirocin and globomycin, the protein inhibitors of IleS and LspA, respectively, were used in sensitization assays of strains containing both asRNA-expressing and open reading frame-expressing plasmids to examine deregulation of the overlapping ileS-lspA genes. We found upstream and downstream polar silencing effects when either ileS or lspA was silenced, indicating coupled expression. Weighted MTL50 values (means and standard deviations) of ribF, ileS, and lspA were 0.65 ± 0.18, 0.64 ± 0.06, and 0.76 ± 0.10, respectively. However, they were not significantly different (P = 0.71 by weighted one-way analysis of variance). The gene requirement for ispH could not be determined due to insufficient growth reduction. Mupirocin and globomycin sensitization experiments indicated that ileS-lspA expression could not be decoupled. The results highlight the inherent challenges associated with genetic analyses of operons; however, coupling of essential genes may provide opportunities to improve RNA-silencing antimicrobials. PMID:26070674

  12. Strategies for Developing Family Nursing Communities of Practice Through Social Media.

    PubMed

    Isaacson, Kris; Looman, Wendy S

    2017-02-01

    This discussion article presents communities of practice (CoPs) and bridging social capital as conceptual frameworks to demonstrate how social media can be leveraged for family nursing knowledge, scholarship, and practice. CoPs require a shared domain of interest, exchange of resources, and dedication to expanding group knowledge. Used strategically and with a professional presence, mainstream social media channels such as Twitter, Facebook, and YouTube can support the family nurse in developing and contributing to CoPs related to family nursing. This article presents four strategies-curate, connect, collaborate, and contribute-for establishing and growing a social media presence that fits one's professional goals and time availability. Family nurses who leverage social media using these strategies can strengthen existing CoPs and at the same time bridge networks to reach new audiences, such as family advocacy groups, policy makers, educators, practitioners, and a wide array of other extended networks.

  13. Dechlorination of lindane by the cyanobacterium Anabaena sp. strain PCC7120 depends on the function of the nir operon.

    PubMed Central

    Kuritz, T; Bocanera, L V; Rivera, N S

    1997-01-01

    Nitrate is essential for lindane dechlorination by the cyanobacteria Anabaena sp. strain PCC7120 and Nostoc ellipsosporum, as it is for dechlorination of other organic compounds by heterotrophic microorganisms. Based on analyses of mutants and effects of environmental factors, we conclude that lindane dechlorination by Anabaena sp. requires a functional nir operon that encodes the enzymes for nitrate utilization. PMID:9150239

  14. Carbon monoxide poisoning: a five year review at Tan Tock Seng Hospital, Singapore.

    PubMed

    Handa, P K; Tai, D Y H

    2005-11-01

    Carbon monoxide poisoning (COP) is one of the leading causes of death from poisoning worldwide. There is no published study of COP in Singapore so far. All patients admitted with the diagnosis of COP to Tan Tock Seng Hospital (TTSH) over 5 years from 1999 to 2003 were retrospectively reviewed. The diagnosis was based on a history of potential exposure to carbon monoxide (CO) and elevated levels of carboxyhaemoglobin (COHb). The causes, demographic data, clinical presentations, management and complications were analysed. There were 12 patients with COP. Their average age was 38.9 (+/-11.8) years, with a male-to-female ratio of 3:1. Accidental COP (58.3%) was more common than intentional COP (41.7%). The most common cause of accidental COP was smoke inhalation from a faulty vehicle. Gas stove was the most preferred source for intentional poisoning. Presenting features were headache (83.3%), confusion (83.3%), coma (12.7%) and agitation (8.3%). The mean COHb level on admission was 35.9% (+/-13.6). All were treated with 100% oxygen. All the patients achieved normal levels of COHb within 24 hours of admission. Two (16.7%) required intubation for airway protection as they were comatose on arrival, of which 1 presented with very high level of COHb (48.1%) and was the only patient to be treated with hyperbaric oxygen. Acute complications were globus pallidus infarction (16.6%), acute respiratory distress syndrome (8.3%) and myocardial ischaemia (8.3%). Most of the patients (91.7%) were discharged well from the hospital. One patient developed parkinsonism after a follow-up of 2 years. There were no deaths. COP is relatively uncommon in Singapore. It has a low rate of short- and long-term complications.

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

    Price, Morgan N.; Arkin, Adam P.; Alm, Eric J.

    Operons are a major feature of all prokaryotic genomes, but how and why operon structures vary is not well understood. To elucidate the life-cycle of operons, we compared gene order between Escherichia coli K12 and its relatives and identified the recently formed and destroyed operons in E. coli. This allowed us to determine how operons form, how they become closely spaced, and how they die. Our findings suggest that operon evolution is driven by selection on gene expression patterns. First, both operon creation and operon destruction lead to large changes in gene expression patterns. For example, the removal of lysAmore » and ruvA from ancestral operons that contained essential genes allowed their expression to respond to lysine levels and DNA damage, respectively. Second, some operons have undergone accelerated evolution, with multiple new genes being added during a brief period. Third, although most operons are closely spaced because of a neutral bias towards deletion and because of selection against large overlaps, highly expressed operons tend to be widely spaced because of regulatory fine-tuning by intervening sequences. Although operon evolution seems to be adaptive, it need not be optimal: new operons often comprise functionally unrelated genes that were already in proximity before the operon formed.« less

  16. Evolutionary dynamics of nematode operons: easy come, slow go.

    PubMed

    Qian, Wenfeng; Zhang, Jianzhi

    2008-03-01

    Operons are widespread in prokaryotes, but are uncommon in eukaryotes, except nematode worms, where approximately 15% of genes reside in over 1100 operons in the model organism Caenorhabditis elegans. It is unclear how operons have become abundant in nematode genomes. The "one-way street" hypothesis asserts that once formed by chance, operons are very difficult to break, because the breakage would leave downstream genes in an operon without a promoter, and hence, unexpressed. To test this hypothesis, we analyzed the presence and absence of C. elegans operons in Caenorhabditis briggsae, Caenorhabditis remanei, and Caenorhabditis brenneri, using Pristionchus pacificus and Brugia malayi as outgroups, and identified numerous operon gains and losses. Coupled with experimental examination of trans-splicing patterns, our comparative genomic analysis revealed diverse molecular mechanisms of operon losses, including inversion, insertion, and relocation, but the presence of internal promoters was not found to facilitate operon losses. In several cases, the data allowed inference of mechanisms by which downstream genes are expressed after operon breakage. We found that the rate of operon gain is approximately 3.3 times that of operon loss. Thus, the evolutionary dynamics of nematode operons is better described as "easy come, slow go," rather than a "one-way street." Based on a mathematic model of operon gains and losses and additional assumptions, we projected that the number of operons in C. elegans will continue to rise by 6%-18% in future evolution before reaching equilibrium between operon gains and losses.

  17. Using a community of practice to evaluate falls prevention activity in a residential aged care organisation: a clinical audit.

    PubMed

    Francis-Coad, Jacqueline; Etherton-Beer, Christopher; Bulsara, Caroline; Nobre, Debbie; Hill, Anne-Marie

    2017-03-01

    Objective This study evaluates whether a community of practice (CoP) could conduct a falls prevention clinical audit and identify gaps in falls prevention practice requiring action. Methods Cross-sectional falls prevention clinical audits were conducted in 13 residential aged care (RAC) sites of a not-for-profit organisation providing care to a total of 779 residents. The audits were led by an operationalised CoP assisted by site clinical staff. A CoP is a group of people with a shared interest who get together to innovate for change. The CoP was made up of self-nominated staff representing all RAC sites and comprised of staff from various disciplines with a shared interest in falls prevention. Results All 13 (100%) sites completed the audit. CoP conduct of the audit met identified criteria for an effective clinical audit. The priorities for improvement were identified as increasing the proportion of residents receiving vitamin D supplementation (mean 41.5%, s.d. 23.7) and development of mandatory falls prevention education for staff and a falls prevention policy, as neither was in place at any site. CoP actions undertaken included a letter to visiting GPs requesting support for vitamin D prescription, surveys of care staff and residents to inform falls education development, defining falls and writing a falls prevention policy. Conclusion A CoP was able to effectively conduct an evidence-based falls prevention activity audit and identify gaps in practice. CoP members were well positioned, as site staff, to overcome barriers and facilitate action in falls prevention practice. What is known about the topic? Audit and feedback is an effective way of measuring clinical quality and safety. CoPs have been established in healthcare using workplace staff to address clinical problems but little is known about their ability to audit and influence practice change. What does this paper add? This study contributes to the body of knowledge on CoPs in healthcare by evaluating the performance of one in the domain of falls prevention audit action. What are the implications for practitioners? A CoP is an effective model to engage staff in the clinical audit process. Clinical audits can raise staff awareness of gaps in practice and motivate staff to plan and action change as recommended in best practice guidelines.

  18. A Homologue of an Operon Required for DNA Transfer in Agrobacterium Is Required in Brucella abortus for Virulence and Intracellular Multiplication

    PubMed Central

    Sieira, Rodrigo; Comerci, Diego J.; Sánchez, Daniel O.; Ugalde, Rodolfo A.

    2000-01-01

    As part of a Brucella abortus 2308 genome project carried out in our laboratory, we identified, cloned, and sequenced a genomic DNA fragment containing a locus (virB) highly homologous to bacterial type IV secretion systems. The B. abortus virB locus is a collinear arrangement of 13 open reading frames (ORFs). Between virB1 and virB2 and downstream of ORF12, two degenerated, palindromic repeat sequences characteristic of Brucella intergenic regions were found. Gene reporter studies demonstrated that the B. abortus virB locus constitutes an operon transcribed from virB1 which is turned on during the stationary phase of growth. A B. abortus polar virB1 mutant failed to replicate in HeLa cells, indicating that the virB operon plays a critical role in intracellular multiplication. Mutants with polar and nonpolar mutations introduced in virB10 showed different behaviors in mice and in the HeLa cell infection assay, suggesting that virB10 per se is necessary for the correct function of this type IV secretion apparatus. Mouse infection assays demonstrated that the virB operon constitutes a major determinant of B. abortus virulence. It is suggested that putative effector molecules secreted by this type IV secretion system determine routing of B. abortus to an endoplasmic reticulum-related replication compartment. PMID:10940027

  19. Biofilm formation and cellulose expression by Bordetella avium 197N, the causative agent of bordetellosis in birds and an opportunistic respiratory pathogen in humans.

    PubMed

    McLaughlin, Kimberley; Folorunso, Ayorinde O; Deeni, Yusuf Y; Foster, Dona; Gorbatiuk, Oksana; Hapca, Simona M; Immoor, Corinna; Koza, Anna; Mohammed, Ibrahim U; Moshynets, Olena; Rogalsky, Sergii; Zawadzki, Kamil; Spiers, Andrew J

    2017-06-01

    Although bacterial cellulose synthase (bcs) operons are widespread within the Proteobacteria phylum, subunits required for the partial-acetylation of the polymer appear to be restricted to a few γ-group soil, plant-associated and phytopathogenic pseudomonads, including Pseudomonas fluorescens SBW25 and several Pseudomonas syringae pathovars. However, a bcs operon with acetylation subunits has also been annotated in the unrelated β-group respiratory pathogen, Bordetella avium 197N. Our comparison of subunit protein sequences and GC content analyses confirms the close similarity between the B. avium 197N and pseudomonad operons and suggests that, in both cases, the cellulose synthase and acetylation subunits were acquired as a single unit. Using static liquid microcosms, we can confirm that B. avium 197N expresses low levels of cellulose in air-liquid interface biofilms and that biofilm strength and attachment levels could be increased by elevating c-di-GMP levels like the pseudomonads, but cellulose was not required for biofilm formation itself. The finding that B. avium 197N is capable of producing cellulose from a highly-conserved, but relatively uncommon bcs operon raises the question of what functional role this modified polymer plays during the infection of the upper respiratory tract or survival between hosts, and what environmental signals control its production. Copyright © 2017 Institut Pasteur. All rights reserved.

  20. A novel determination of energy expenditure efficiency during a balance task using accelerometers. A pilot study.

    PubMed

    Caña-Pino, Alejandro; Apolo-Arenas, Maria Dolores; Moral-Blanco, Javier; De la Cruz-Sánchez, Ernesto; Espejo-Antúnez, Luis

    2017-08-18

    The objectives of this study are to determine the displacement of the center of pressure (CoP) and its association with the spectral energy density of the acceleration required for the maintenance of postural balance in different standing positions in healthy participants using design observational and setting laboratorial studies. Participants were 30 healthy university students aged between 18 and 32 years old (mean [M] ± standard deviation [SD] = 21,57 ± 3,31 years). Triaxial accelerometer and a pressure platform were used in order to obtain energy spectral density and CoP sway measurements during four balance tasks. Statistically significant differences were found for anteroposterior (p = 0.002) and mediolateral (p = 0.009) CoP displacement between the conditions eyes closed and stable surface and the conditions eyes closed and unstable surface. A statistically significant correlation was also observed between Z-axis (anterior-posterior) of the accelerometer and mediolateral axis of the CoP (r = 0.465; p = 0.01) and between Y-axis accelerometer (mediolateral) and displacement of the CoP in the anteroposterior axis (r = 0.413; p = 0.023). Spectral energy density appears to be associated with the displacement of CoP in healthy participants.

  1. Myeloid leukemia factor 1 regulates p53 by suppressing COP1 via COP9 signalosome subunit 3

    PubMed Central

    Yoneda-Kato, Noriko; Tomoda, Kiichiro; Umehara, Mari; Arata, Yukinobu; Kato, Jun-ya

    2005-01-01

    Myeloid leukemia factor 1 (MLF1) was first identified as the leukemic fusion protein NPM-MLF1 generated by the t(3;5)(q25.1;q34) chromosomal translocation. Although MLF1 expresses normally in a variety of tissues including hematopoietic stem cells and the overexpression of MLF1 correlates with malignant transformation in human cancer, little is known about how MLF1 is involved in the regulation of cell growth. Here we show that MLF1 is a negative regulator of cell cycle progression functioning upstream of the tumor suppressor p53. MLF1 induces p53-dependent cell cycle arrest in murine embryonic fibroblasts. This action requires a novel binding partner, subunit 3 of the COP9 signalosome (CSN3). A reduction in the level of CSN3 protein with small interfering RNA abrogated MLF1-induced G1 arrest and impaired the activation of p53 by genotoxic stress. Furthermore, ectopic MLF1 expression and CSN3 knockdown inversely affect the endogenous level of COP1, a ubiquitin ligase for p53. Exogenous expression of COP1 overcomes MLF1-induced growth arrest. These results indicate that MLF1 is a critical regulator of p53 and suggest its involvement in leukemogenesis through a novel CSN3–COP1 pathway. PMID:15861129

  2. The Bacillus subtilis GntR family repressor YtrA responds to cell wall antibiotics.

    PubMed

    Salzberg, Letal I; Luo, Yun; Hachmann, Anna-Barbara; Mascher, Thorsten; Helmann, John D

    2011-10-01

    The transglycosylation step of cell wall synthesis is a prime antibiotic target because it is essential and specific to bacteria. Two antibiotics, ramoplanin and moenomycin, target this step by binding to the substrate lipid II and the transglycosylase enzyme, respectively. Here, we compare the ramoplanin and moenomycin stimulons in the Gram-positive model organism Bacillus subtilis. Ramoplanin strongly induces the LiaRS two-component regulatory system, while moenomycin almost exclusively induces genes that are part of the regulon of the extracytoplasmic function (ECF) σ factor σ(M). Ramoplanin additionally induces the ytrABCDEF and ywoBCD operons, which are not part of a previously characterized antibiotic-responsive regulon. Cluster analysis reveals that these two operons are selectively induced by a subset of cell wall antibiotics that inhibit lipid II function or recycling. Repression of both operons requires YtrA, which recognizes an inverted repeat in front of its own operon and in front of ywoB. These results suggest that YtrA is an additional regulator of cell envelope stress responses.

  3. The Bacillus subtilis GntR Family Repressor YtrA Responds to Cell Wall Antibiotics▿§

    PubMed Central

    Salzberg, Letal I.; Luo, Yun; Hachmann, Anna-Barbara; Mascher, Thorsten; Helmann, John D.

    2011-01-01

    The transglycosylation step of cell wall synthesis is a prime antibiotic target because it is essential and specific to bacteria. Two antibiotics, ramoplanin and moenomycin, target this step by binding to the substrate lipid II and the transglycosylase enzyme, respectively. Here, we compare the ramoplanin and moenomycin stimulons in the Gram-positive model organism Bacillus subtilis. Ramoplanin strongly induces the LiaRS two-component regulatory system, while moenomycin almost exclusively induces genes that are part of the regulon of the extracytoplasmic function (ECF) σ factor σM. Ramoplanin additionally induces the ytrABCDEF and ywoBCD operons, which are not part of a previously characterized antibiotic-responsive regulon. Cluster analysis reveals that these two operons are selectively induced by a subset of cell wall antibiotics that inhibit lipid II function or recycling. Repression of both operons requires YtrA, which recognizes an inverted repeat in front of its own operon and in front of ywoB. These results suggest that YtrA is an additional regulator of cell envelope stress responses. PMID:21856850

  4. InfoSec-MobCop - Framework for Theft Detection and Data Security on Mobile Computing Devices

    NASA Astrophysics Data System (ADS)

    Gupta, Anand; Gupta, Deepank; Gupta, Nidhi

    People steal mobile devices with the intention of making money either by selling the mobile or by taking the sensitive information stored inside it. Mobile thefts are rising even with existing deterrents in place. This is because; they are ineffective, as they generate unnecessary alerts and might require expensive hardware equipments. In this paper a novel framework termed as InfoSec-MobCop is proposed which secures a mobile user’s data and discovers theft by detecting any anomaly in the user behavior. The anomaly of the user is computed by extracting and monitoring user specific details (typing pattern and usage history). The result of any intrusion attempt by a masquerader is intimated to the service provider through an SMS. Effectiveness of the used approach is discussed using FAR and FRR graphs. The experimental system uses both real users and simulated studies to quantify the effectiveness of the InfoSec-MobCop (Information Security Mobile Cop).

  5. A global analysis of adaptive evolution of operons in cyanobacteria.

    PubMed

    Memon, Danish; Singh, Abhay K; Pakrasi, Himadri B; Wangikar, Pramod P

    2013-02-01

    Operons are an important feature of prokaryotic genomes. Evolution of operons is hypothesized to be adaptive and has contributed significantly towards coordinated optimization of functions. Two conflicting theories, based on (i) in situ formation to achieve co-regulation and (ii) horizontal gene transfer of functionally linked gene clusters, are generally considered to explain why and how operons have evolved. Furthermore, effects of operon evolution on genomic traits such as intergenic spacing, operon size and co-regulation are relatively less explored. Based on the conservation level in a set of diverse prokaryotes, we categorize the operonic gene pair associations and in turn the operons as ancient and recently formed. This allowed us to perform a detailed analysis of operonic structure in cyanobacteria, a morphologically and physiologically diverse group of photoautotrophs. Clustering based on operon conservation showed significant similarity with the 16S rRNA-based phylogeny, which groups the cyanobacterial strains into three clades. Clade C, dominated by strains that are believed to have undergone genome reduction, shows a larger fraction of operonic genes that are tightly packed in larger sized operons. Ancient operons are in general larger, more tightly packed, better optimized for co-regulation and part of key cellular processes. A sub-clade within Clade B, which includes Synechocystis sp. PCC 6803, shows a reverse trend in intergenic spacing. Our results suggest that while in situ formation and vertical descent may be a dominant mechanism of operon evolution in cyanobacteria, optimization of intergenic spacing and co-regulation are part of an ongoing process in the life-cycle of operons.

  6. Detecting uber-operons in prokaryotic genomes.

    PubMed

    Che, Dongsheng; Li, Guojun; Mao, Fenglou; Wu, Hongwei; Xu, Ying

    2006-01-01

    We present a study on computational identification of uber-operons in a prokaryotic genome, each of which represents a group of operons that are evolutionarily or functionally associated through operons in other (reference) genomes. Uber-operons represent a rich set of footprints of operon evolution, whose full utilization could lead to new and more powerful tools for elucidation of biological pathways and networks than what operons have provided, and a better understanding of prokaryotic genome structures and evolution. Our prediction algorithm predicts uber-operons through identifying groups of functionally or transcriptionally related operons, whose gene sets are conserved across the target and multiple reference genomes. Using this algorithm, we have predicted uber-operons for each of a group of 91 genomes, using the other 90 genomes as references. In particular, we predicted 158 uber-operons in Escherichia coli K12 covering 1830 genes, and found that many of the uber-operons correspond to parts of known regulons or biological pathways or are involved in highly related biological processes based on their Gene Ontology (GO) assignments. For some of the predicted uber-operons that are not parts of known regulons or pathways, our analyses indicate that their genes are highly likely to work together in the same biological processes, suggesting the possibility of new regulons and pathways. We believe that our uber-operon prediction provides a highly useful capability and a rich information source for elucidation of complex biological processes, such as pathways in microbes. All the prediction results are available at our Uber-Operon Database: http://csbl.bmb.uga.edu/uber, the first of its kind.

  7. Detecting uber-operons in prokaryotic genomes

    PubMed Central

    Che, Dongsheng; Li, Guojun; Mao, Fenglou; Wu, Hongwei; Xu, Ying

    2006-01-01

    We present a study on computational identification of uber-operons in a prokaryotic genome, each of which represents a group of operons that are evolutionarily or functionally associated through operons in other (reference) genomes. Uber-operons represent a rich set of footprints of operon evolution, whose full utilization could lead to new and more powerful tools for elucidation of biological pathways and networks than what operons have provided, and a better understanding of prokaryotic genome structures and evolution. Our prediction algorithm predicts uber-operons through identifying groups of functionally or transcriptionally related operons, whose gene sets are conserved across the target and multiple reference genomes. Using this algorithm, we have predicted uber-operons for each of a group of 91 genomes, using the other 90 genomes as references. In particular, we predicted 158 uber-operons in Escherichia coli K12 covering 1830 genes, and found that many of the uber-operons correspond to parts of known regulons or biological pathways or are involved in highly related biological processes based on their Gene Ontology (GO) assignments. For some of the predicted uber-operons that are not parts of known regulons or pathways, our analyses indicate that their genes are highly likely to work together in the same biological processes, suggesting the possibility of new regulons and pathways. We believe that our uber-operon prediction provides a highly useful capability and a rich information source for elucidation of complex biological processes, such as pathways in microbes. All the prediction results are available at our Uber-Operon Database: , the first of its kind. PMID:16682449

  8. Examining Changes to Center of Pressure During the First Trials of Wii Gameplay.

    PubMed

    Reed-Jones, Rebecca; Carvalho, Laura; Sanderson, Chelsey; Montelpare, William; Murray, Nicholas; Powell, Douglas

    2017-02-01

    Use of the Nintendo Wii™ as a balance assessment and rehabilitation tool continues to grow. One advantage of the Wii is that games can serve as a virtual reality training tool; however, a disadvantage of the Wii is the human-machine interface and the learning effect over multiple trials. The purpose of this study was to assess changes in postural control during Wii gameplay over a series of trials in novice players. Thirty-one university athletes (aged 18-25 years) completed four trials of the Nintendo Wii Fit™ soccer heading (SH) balance game. Center of pressure (COP) was calculated in the anterior-posterior (AP) and medial-lateral (ML) directions for each 70-second time trial at 1000 Hz. COP was assessed using six linear and two nonlinear measures. Repeated measures analysis of variances compared COP measures over the four trials. Significant differences in COP magnitude and velocity were found between trials 1 and 2 in the ML direction. No significant effects of trial were found in the AP direction. In contrast, a measure of the overall area of COP using an ellipse method revealed a significant reduction to COP area between trials 3 and 4. No significant differences between trials were observed in nonlinear measures. These results demonstrate how magnitude and velocity measures of COP control stabilize after the first trial of Wii SH game play in novice young adults. As Wii rehabilitation focuses on individuals with balance difficulties, an important consideration when using the game as an assessment tool is that more than four trials may be required to capture learning in these populations. In addition, contrasting results from ellipse measurement methods point to the use of multiple measures for robust description of COP behavior. This work provides understanding of normative postural control responses with further research in clinical populations needed.

  9. Robust C–C bonded porous networks with chemically designed functionalities for improved CO2 capture from flue gas

    PubMed Central

    Thirion, Damien; Lee, Joo S; Özdemir, Ercan

    2016-01-01

    Effective carbon dioxide (CO2) capture requires solid, porous sorbents with chemically and thermally stable frameworks. Herein, we report two new carbon–carbon bonded porous networks that were synthesized through metal-free Knoevenagel nitrile–aldol condensation, namely the covalent organic polymer, COP-156 and 157. COP-156, due to high specific surface area (650 m2/g) and easily interchangeable nitrile groups, was modified post-synthetically into free amine- or amidoxime-containing networks. The modified COP-156-amine showed fast and increased CO2 uptake under simulated moist flue gas conditions compared to the starting network and usual industrial CO2 solvents, reaching up to 7.8 wt % uptake at 40 °C. PMID:28144294

  10. Relationship of the functional movement screen in-line lunge to power, speed, and balance measures.

    PubMed

    Hartigan, Erin H; Lawrence, Michael; Bisson, Brian M; Torgerson, Erik; Knight, Ryan C

    2014-05-01

    The in-line lunge of the Functional Movement Screen (FMS) evaluates lateral stability, balance, and movement asymmetries. Athletes who score poorly on the in-line lunge should avoid activities requiring power or speed until scores are improved, yet relationships between the in-line lunge scores and other measures of balance, power, and speed are unknown. (1) Lunge scores will correlate with center of pressure (COP), maximum jump height (MJH), and 36.6-meter sprint time and (2) there will be no differences between limbs on lunge scores, MJH, or COP. Descriptive laboratory study. Level 3. Thirty-seven healthy, active participants completed the first 3 tasks of the FMS (eg, deep squat, hurdle step, in-line lunge), unilateral drop jumps, and 36.6-meter sprints. A 3-dimensional motion analysis system captured MJH. Force platforms measured COP excursion. A laser timing system measured 36.6-m sprint time. Statistical analyses were used to determine whether a relationship existed between lunge scores and COP, MJH, and 36.6-m speed (Spearman rho tests) and whether differences existed between limbs in lunge scores (Wilcoxon signed-rank test), MJH, and COP (paired t tests). Lunge scores were not significantly correlated with COP, MJH, or 36.6-m sprint time. Lunge scores, COP excursion, and MJH were not statistically different between limbs. Performance on the FMS in-line lunge was not related to balance, power, or speed. Healthy participants were symmetrical in lunging measures and MJH. Scores on the FMS in-line lunge should not be attributed to power, speed, or balance performance without further examination. However, assessing limb symmetry appears to be clinically relevant.

  11. Improved Analytical Method for Determination of Cholesterol-Oxidation Products in Meat and Animal Fat by QuEChERS Coupled with Gas Chromatography-Mass Spectrometry.

    PubMed

    Chiu, Che-Wei; Kao, Tsai-Hua; Chen, Bing-Huei

    2018-04-04

    Cholesterol is widely present in animal fats and meat products and can undergo oxidation to form cholesterol-oxidation products (COPs) during heating. The objective of this study was to develop a QuEChERS method for the determination of COPs in edible animal fats and meat products via gas chromatography-mass spectrometry in which the required solvent volume and extraction time were reduced. By employing a DB-5MS capillary column (30 m × 0.25 mm i.d., 0.25 μm film thickness) and a temperature-programming method, seven COPs, cholesterol, and the internal standard 5α-cholestane could be separated within 19 min. The limits of detection and limits of quantitation based on the COP standards ranged from 0.16 to 180 ng/mL and from 0.32 to 400 ng/mL, respectively, and the recoveries ranged from 89.1 to 107.6% for boiled pork and from 80.5 to 105.6% for lard. The intraday variabilities for boiled pork and lard ranged from 2.27 to 6.87% and from 1.52 to 9.78%, respectively, whereas the interday variabilities ranged from 1.81 to 7.89% and from 3.57 to 9.26%, respectively. Among the various meat samples, fish showed the highest level of COPs (31.84 μg/g). For the edible fats, the COP contents in tallow (22.79-60.15 μg/g) were much higher than those in lard (0.152-2.55 μg/g) and butter (0.526-1.36 μg/g). Collectively, this method can be applied to determine COPs in cholesterol-containing foodstuffs.

  12. The mechanism of Cu+ transport ATPases: interaction with CU+ chaperones and the role of transient metal-binding sites.

    PubMed

    Padilla-Benavides, Teresita; McCann, Courtney J; Argüello, José M

    2013-01-04

    Cu(+)-ATPases are membrane proteins that couple the hydrolysis of ATP to the efflux of cytoplasmic Cu(+). In cells, soluble chaperone proteins bind and distribute cytoplasmic Cu(+), delivering the ion to the transmembrane metal-binding sites in the ATPase. The structure of Legionella pneumophila Cu(+)-ATPase (Gourdon, P., Liu, X. Y., Skjørringe, T., Morth, J. P., Møller, L. B., Pedersen, B. P., and Nissen, P. (2011) Nature 475, 59-64) shows that a kinked transmembrane segment forms a "platform" exposed to the cytoplasm. In addition, neighboring invariant Met, Asp, and Glu are located at the "entrance" of the ion path. Mutations of amino acids in these regions of the Archaeoglobus fulgidus Cu(+)-ATPase CopA do not affect ATPase activity in the presence of Cu(+) free in solution. However, Cu(+) bound to the corresponding chaperone (CopZ) could not activate the mutated ATPases, and in parallel experiments, CopZ was unable to transfer Cu(+) to CopA. Furthermore, mutation of a specific electronegative patch on the CopZ surface abolishes the ATPase activation and Cu(+) transference, indicating that the region is required for the CopZ-CopA interaction. Moreover, the data suggest that the interaction is driven by the complementation of the electropositive platform in the ATPase and the electronegative Cu(+) chaperone. This docking likely places the Cu(+) proximal to the conserved carboxyl and thiol groups in the entrance site that induce metal release from the chaperone via ligand exchange. The initial interaction of Cu(+) with the pump is transient because Cu(+) is transferred from the entrance site to transmembrane metal-binding sites involved in transmembrane translocation.

  13. Reliability of center of pressure measures within and between sessions in individuals post-stroke and healthy controls.

    PubMed

    Gray, Vicki L; Ivanova, Tanya D; Garland, S Jayne

    2014-01-01

    Knowing the reliability of the center of pressure (COP) is important for interpreting balance deficits post-stroke, especially when the balance deficits can necessitate the use of short duration trials. The novel aspect of this reliability study was to examine the center of pressure measures using two adjacent force platforms between and within sessions in stroke and controls. After stroke, it is important to understand the contribution of the paretic and non-paretic leg to the motor control of standing balance. Because there is a considerable body of knowledge on COP reliability on a single platform, we chose to examine reliability using two adjacent platforms which has not been examined previously in stroke. Twenty participants post-stroke and 22 controls performed an arm raise, load drop and quiet stance balance task while standing on two adjacent force platforms, on two separate days. Intraclass correlations coefficient (ICC2,1) and percentage standard error of measurement (SEM%) were calculated for COP velocity, ellipse area, anterior-posterior (AP) displacement, and medial-lateral (ML) displacement. Between sessions, COP velocity was the most reliable with high ICCs and low SEM% across groups and tasks and ellipse area was less reliable with low ICCs across groups and tasks. COP measures were less reliable during the arm raise than load drop post-stroke. Within session reliability was high for COP velocity and ML displacement requiring no more than six trials across tasks. The COP velocity was the most reliable measure with high ICCs between sessions and the high reliability was achieved with fewer trials in both groups in a single session. Copyright © 2014 Elsevier B.V. All rights reserved.

  14. The Copper Homeostasis Transcription Factor CopR Is Involved in H2O2 Stress in Lactobacillus plantarum CAUH2

    PubMed Central

    Yang, Yang; Yin, Jia; Liu, Jie; Xu, Qi; Lan, Tian; Ren, Fazheng; Hao, Yanling

    2017-01-01

    Transcriptional factors (TFs) play important roles in the responses to oxidative, acid, and other environmental stresses in Gram-positive bacteria, but the regulatory mechanism of TFs involved in oxidative stress remains unknown in lactic acid bacteria. In the present work, homologous overexpression strains with 43 TFs were constructed in the Lactobacillus plantarum CAUH2 parent strain. The strain overexpressing CopR displayed the highest sensitivity and a 110-fold decrease in survival rate under H2O2 challenge. The importance of CopR in the response to H2O2 stress was further confirmed by a 10.8-fold increase in the survival of a copR insertion mutant. In silico analysis of the genes flanking copR revealed putative CopR-binding “cop box” sequences in the promoter region of the adjacent gene copB encoding a Cu2+-exporting ATPase. Electrophoretic mobility shift assay (EMSA) analysis demonstrated the specific binding of CopR with copB in vitro, suggesting copB is a target gene of CopR in L. plantarum. The role of CopB involved in oxidative stress was verified by the significantly decreased survival in the copB mutant. Furthermore, a growth defect in copper-containing medium demonstrated that CopB functions as an export ATPase for copper ions. Furthermore, EMSAs revealed that CopR functions as a regulator that negatively regulates copB gene and Cu2+ serves as inducer of CopR to activate the expression of CopB in response to H2O2 stress in L. plantarum CAUH2. Our findings indicated that CopR plays an important role in enhancing oxidative resistance by regulating copB to modulate copper homeostasis. PMID:29089937

  15. Postural control and head stability during natural gaze behaviour in 6- to 12-year-old children.

    PubMed

    Schärli, A M; van de Langenberg, R; Murer, K; Müller, R M

    2013-06-01

    We investigated how the influence of natural exploratory gaze behaviour on postural control develops from childhood into adulthood. In a cross-sectional design, we compared four age groups: 6-, 9-, 12-year-olds and young adults. Two experimental trials were performed: quiet stance with a fixed gaze (fixed) and quiet stance with natural exploratory gaze behaviour (exploratory). The latter was elicited by having participants watch an animated short film on a large screen in front of them. 3D head rotations in space and centre of pressure (COP) excursions on the ground plane were measured. Across conditions, both head rotation and COP displacement decreased with increasing age. Head movement was greater in the exploratory condition in all age groups. In all children-but not in adults-COP displacement was markedly greater in the exploratory condition. Bivariate correlations across groups showed highly significant positive correlations between COP displacement in ML direction and head rotation in yaw, roll, and pitch in both conditions. The regularity of COP displacements did not show a clear developmental trend, which indicates that COP dynamics were qualitatively similar across age groups. Together, the results suggest that the contribution of head movement to eye-head saccades decreases with age and that head instability-in part resulting from such gaze-related head movements-is an important limiting factor in children's postural control. The lack of head stabilisation might particularly affect children in everyday activities in which both postural control and visual exploration are required.

  16. Operon-mapper: A Web Server for Precise Operon Identification in Bacterial and Archaeal Genomes.

    PubMed

    Taboada, Blanca; Estrada, Karel; Ciria, Ricardo; Merino, Enrique

    2018-06-19

    Operon-mapper is a web server that accurately, easily, and directly predicts the operons of any bacterial or archaeal genome sequence. The operon predictions are based on the intergenic distance of neighboring genes as well as the functional relationships of their protein-coding products. To this end, Operon-mapper finds all the ORFs within a given nucleotide sequence, along with their genomic coordinates, orthology groups, and functional relationships. We believe that Operon-mapper, due to its accuracy, simplicity and speed, as well as the relevant information that it generates, will be a useful tool for annotating and characterizing genomic sequences. http://biocomputo.ibt.unam.mx/operon_mapper/.

  17. Arabidopsis cop8 and fus4 mutations define the same gene that encodes subunit 4 of the COP9 signalosome.

    PubMed Central

    Serino, G; Tsuge, T; Kwok, S; Matsui, M; Wei, N; Deng, X W

    1999-01-01

    The pleiotropic constitutive photomorphogenic/deetiolated/fusca (cop/det/fus) mutants of Arabidopsis exhibit features of light-grown seedlings when grown in the dark. Cloning and biochemical analysis of COP9 have revealed that it is a component of a multiprotein complex, the COP9 signalosome (previously known as the COP9 complex). Here, we compare the immunoaffinity and the biochemical purification of the COP9 signalosome from cauliflower and confirm its eight-subunit composition. Molecular cloning of subunit 4 of the complex revealed that it is a proteasome-COP9 complex-eIF3 domain protein encoded by a gene that maps to chromosome 5, near the chromosomal location of the cop8 and fus4 mutations. Genetic complementation tests showed that the cop8 and fus4 mutations define the same locus, now designated as COP8. Molecular analysis of the subunit 4-encoding gene in both cop8 and fus4 mutants identified specific molecular lesions, and overexpression of the subunit 4 cDNA in a cop8 mutant background resulted in complete rescue of the mutant phenotype. Thus, we conclude that COP8 encodes subunit 4 of the COP9 signalosome. Examination of possible molecular interactions by using the yeast two-hybrid assay indicated that COP8 is capable of strong self-association as well as interaction with COP9, FUS6/COP11, FUS5, and Arabidopsis JAB1 homolog 1, the latter four proteins being previously defined subunits of the Arabidopsis COP9 signalosome. A comparative sequence analysis indicated that COP8 is highly conserved among multicellular eukaryotes and is also similar to a subunit of the 19S regulatory particle of the 26S proteasome. PMID:10521526

  18. Phenotypic and Molecular Analysis of Mes-3, a Maternal-Effect Gene Required for Proliferation and Viability of the Germ Line in C. Elegans

    PubMed Central

    Paulsen, J. E.; Capowski, E. E.; Strome, S.

    1995-01-01

    mes-3 is one of four maternal-effect sterile genes that encode maternal components required for normal postembryonic development of the germ line in Caenorhabditis elegans. mes-3 mutant mothers produce sterile progeny, which contain few germ cells and no gametes. This terminal phenotype reflects two problems: reduced proliferation of the germ line and germ cell death. Both the appearance of the dying germ cells and the results of genetic tests indicate that germ cells in mes-3 animals undergo a necrotic-like death, not programmed cell death. The few germ cells that appear healthy in mes-3 worms do not differentiate into gametes, even after elimination of the signaling pathway that normally maintains the undifferentiated population of germ cells. Thus, mes-3 encodes a maternally supplied product that is required both for proliferation of the germ line and for maintenance of viable germ cells that are competent to differentiate into gametes. Cloning and molecular characterization of mes-3 revealed that it is the upstream gene in an operon. The genes in the operon display parallel expression patterns; transcripts are present throughout development and are not restricted to germ-line tissue. Both mes-3 and the downstream gene in the operon encode novel proteins. PMID:8601481

  19. Two Paralogous Families of a Two-Gene Subtilisin Operon Are Widely Distributed in Oral Treponemes

    PubMed Central

    Correia, Frederick F.; Plummer, Alvin R.; Ellen, Richard P.; Wyss, Chris; Boches, Susan K.; Galvin, Jamie L.; Paster, Bruce J.; Dewhirst, Floyd E.

    2003-01-01

    Certain oral treponemes express a highly proteolytic phenotype and have been associated with periodontal diseases. The periodontal pathogen Treponema denticola produces dentilisin, a serine protease of the subtilisin family. The two-gene operon prcA-prtP is required for expression of active dentilisin (PrtP), a putative lipoprotein attached to the treponeme's outer membrane or sheath. The purpose of this study was to examine the diversity and structure of treponemal subtilisin-like proteases in order to better understand their distribution and function. The complete sequences of five prcA-prtP operons were determined for Treponema lecithinolyticum, “Treponema vincentii,” and two canine species. Partial operon sequences were obtained for T. socranskii subsp. 04 as well as 450- to 1,000-base fragments of prtP genes from four additional treponeme strains. Phylogenetic analysis demonstrated that the sequences fall into two paralogous families. The first family includes the sequence from T. denticola. Treponemes possessing this operon family express chymotrypsin-like protease activity and can cleave the substrate N-succinyl-alanyl-alanyl-prolyl-phenylalanine-p-nitroanilide (SAAPFNA). Treponemes possessing the second paralog family do not possess chymotrypsin-like activity or cleave SAAPFNA. Despite examination of a range of protein and peptide substrates, the specificity of the second protease family remains unknown. Each of the fully sequenced prcA and prtP genes contains a 5′ hydrophobic leader sequence with a treponeme lipobox. The two paralogous families of treponeme subtilisins represent a new subgroup within the subtilisin family of proteases and are the only subtilisin lipoprotein family. The present study demonstrated that the subtilisin paralogs comprising a two-gene operon are widely distributed among treponemes. PMID:14617650

  20. Toilet Grab-Bar Preference and Center of Pressure Deviation During Toilet Transfers in Healthy Seniors, Seniors With Hip Replacements, and Seniors Having Suffered a Stroke.

    PubMed

    Kennedy, Matthew Joel; Arcelus, Amaya; Guitard, Paulette; Goubran, R A; Sveistrup, Heidi

    2015-01-01

    Multiple toilet grab-bar configurations are required by people with a diverse spectrum of disability. The study purpose was to determine toilet grab-bar preference of healthy seniors, seniors with a hip replacement, and seniors post-stroke, and to determine the effect of each configuration on centre of pressure (COP) displacement during toilet transfers. 14 healthy seniors, 7 ambulatory seniors with a hip replacement, and 8 ambulatory seniors post-stroke participated in the study. Toilet transfers were performed with no bars (NB), commode (C), two vertical bars (2VB), one vertical bar (1VB), a horizontal bar (H), two swing-away bars (S) and a diagonal bar (D). COP was measured using pressure sensitive floor mats. Participants rated the safety, ease of use, helpfulness, comfort and preference for instalment. 2VB was most preferred and had the smallest COP deviation. Least preferred was H and NB. C caused largest COP displacement but had favourable ratings. The preference and safety of the 2VB should be considered in the design of accessible toilets and in accessibility construction guidelines. However these results need to be verified in non-ambulatory populations. C is frequently prescribed, but generates large COP deviation, suggesting it may present an increased risk of falls.

  1. Characteristics and bioactivities of different molecular weight polysaccharides from camellia seed cake.

    PubMed

    Xu, Zhou; Li, Xu; Feng, Shiling; Liu, Jing; Zhou, Lijun; Yuan, Ming; Ding, Chunbang

    2016-10-01

    Four polysaccharides, namely COP-1, COP-2, COP-3 and COP-4, were ultrafiltrated from crud Camellia oleifera seed cake polysaccharides (COP-c), purified, and characterized, including the determination of antioxidant and antiproliferative activities. Their molecular weights were 7.9, 36, 83 and 225kDa, respectively. All COPs showed the similar FT-IR spectrums, but significant differentials in monosaccharide components. COP-2 exhibited the highest radical scavenging abilities. COP-1 has the strongest metal chelating capabilities. Although with higher molecular weight, COP-4 showed the poorest antioxidant abilities. These results suggested appreciate molecular weight COP possessed a better antioxidant activities. Additionally, all COPs had non-significant antiproliferative abilities in HaLa and HepG2 cells. Copyright © 2016 Elsevier B.V. All rights reserved.

  2. Age, gender, and percentage of circulating osteoprogenitor (COP) cells: The COP Study.

    PubMed

    Gunawardene, Piumali; Al Saedi, Ahmed; Singh, Lakshman; Bermeo, Sandra; Vogrin, Sara; Phu, Steven; Suriyaarachchi, Pushpa; Pignolo, Robert J; Duque, Gustavo

    2017-10-01

    Circulating osteoprogenitor (COP) cells are blood-borne cells which express a variety of osteoblastic markers and are able to form bone nodules in vivo. Whereas a high percentage of COP cells (%COP) is associated with vascular calcification, low %COP has been associated with disability and frailty. However, the reference range of %COP in age- and gender-matching populations, and the age-related changes in %COP remain unknown. A cross-sectional study was undertaken in 144 healthy volunteers in Western Sydney (20-90year-old, 10 male and 10 female subjects per decade). %COP was quantified by flow cytometry. A high inter-and intra-rater reliability was found. In average, in this healthy population average of %COP was 0.42. There was no significant difference in %COP among the age groups. Similarly, no significant difference was found in %COP with gender, weight, height or BMI. In addition, we identified a normal reference range of %COP of 0.1-3.8%. In conclusion, in addition to the identification of steady levels of COP cells with age, we also identified a normal reference range of %COP, which could be used in future studies looking at musculoskeletal diseases in older populations. Copyright © 2017 Elsevier Inc. All rights reserved.

  3. Bistable behavior of the lac operon in E. coli when induced with a mixture of lactose and TMG.

    PubMed

    Díaz-Hernández, Orlando; Santillán, Moisés

    2010-01-01

    In this work we investigate multistability in the lac operon of Escherichia coli when it is induced by a mixture of lactose and the non-metabolizable thiomethyl galactoside (TMG). In accordance with previously published experimental results and computer simulations, our simulations predict that: (1) when the system is induced by TMG, the system shows a discernible bistable behavior while, (2) when the system is induced by lactose, bistability does not disappear but excessively high concentrations of lactose would be required to observe it. Finally, our simulation results predict that when a mixture of lactose and TMG is used, the bistability region in the extracellular glucose concentration vs. extracellular lactose concentration parameter space changes in such a way that the model predictions regarding bistability could be tested experimentally. These experiments could help to solve a recent controversy regarding the existence of bistability in the lac operon under natural conditions.

  4. Evidence against the selfish operon theory.

    PubMed

    Pál, Csaba; Hurst, Laurence D

    2004-06-01

    According to the selfish operon hypothesis, the clustering of genes and their subsequent organization into operons is beneficial for the constituent genes because it enables the horizontal gene transfer of weakly selected, functionally coupled genes. The majority of these are expected to be non-essential genes. From our analysis of the Escherichia coli genome, we conclude that the selfish operon hypothesis is unlikely to provide a general explanation for clustering nor can it account for the gene composition of operons. Contrary to expectations, essential genes with related functions have an especially strong tendency to cluster, even if they are not in operons. Moreover, essential genes are particularly abundant in operons.

  5. Comparison of whole blood and plasma colloid osmotic pressure in healthy cats.

    PubMed

    Jackson, Mary L; Kerl, Marie E; Tynan, Beth; Mann, F A

    2014-01-01

    To establish reference intervals for whole blood and plasma colloid osmotic pressure (COP) in healthy cats between the ages of 1 and 10 years using a cage-side colloid osmometer. Prospective, observational study. University veterinary teaching hospital. Sixty-three healthy cats. Phlebotomy. Whole blood COP mean was 24.4 (±2.78) mmHg and plasma COP mean was 24.3 (±2.59) mmHg. Reference interval for our study population of feline whole blood COP was 18.9 to 30.4 mmHg, and for our study population of feline plasma COP was 18.3 to 30.8 mmHg. Difference of paired whole blood COP and plasma COP was +0.23 ± 1.68 mmHg (P = 0.32). There was no significant difference when comparing COP from neutered male and neutered female cats. Total protein and albumin were significantly correlated with whole blood COP (total protein to whole blood COP P < 0.0001, r = 0.53; albumin to whole blood COP P <0.0001, r = 0.68) and plasma COP (total protein to plasma COP P = 0.0025, r = 0.41; albumin to plasma COP P < 0.0001, r = 0.66). No significant difference was found between mean whole blood and plasma COP in this study population of cats. Even though not statistically significant, evaluation of paired whole blood COP and plasma COP did reveal a slight difference; therefore, it seems prudent to maintain sample consistency for serial evaluations in cats. © Veterinary Emergency and Critical Care Society 2014.

  6. Coefficient of performance for a low-dissipation Carnot-like refrigerator with nonadiabatic dissipation

    NASA Astrophysics Data System (ADS)

    Hu, Yong; Wu, Feifei; Ma, Yongli; He, Jizhou; Wang, Jianhui; Hernández, A. Calvo; Roco, J. M. M.

    2013-12-01

    We study the coefficient of performance (COP) and its bounds for a Carnot-like refrigerator working between two heat reservoirs at constant temperatures Th and Tc, under two optimization criteria χ and Ω. In view of the fact that an “adiabatic” process usually takes finite time and is nonisentropic, the nonadiabatic dissipation and the finite time required for the adiabatic processes are taken into account by assuming low dissipation. For given optimization criteria, we find that the lower and upper bounds of the COP are the same as the corresponding ones obtained from the previous idealized models where any adiabatic process is undergone instantaneously with constant entropy. To describe some particular models with very fast adiabatic transitions, we also consider the influence of the nonadiabatic dissipation on the bounds of the COP, under the assumption that the irreversible entropy production in the adiabatic process is constant and independent of time. Our theoretical predictions match the observed COPs of real refrigerators more closely than the ones derived in the previous models, providing a strong argument in favor of our approach.

  7. Getting to uptake: do communities of practice support the implementation of evidence-based practice?

    PubMed

    Barwick, Melanie A; Peters, Julia; Boydell, Katherine

    2009-02-01

    Practitioners are increasingly encouraged to adopt evidence-based practices (EBP) leading to a need for new knowledge translation strategies to support implementation and practice change. This study examined the benefits of a community of practice in the context of Ontario's children's mental health sector where organizations are mandated to adopt a standardized outcome measure to monitor client response to treatment. Readiness for change, practice change, content knowledge, and satisfaction with and use of implementation supports were examined among practitioners newly trained on the measure who were randomly assigned to a community of practice (CoP) or a practice as usual (PaU) group. CoP practitioners attended 6 sessions over 12 months; PaU practitioners had access to usual implementation supports. Groups did not differ on readiness for change or reported practice change, although CoP participants demonstrated greater use of the tool in practice, better content knowledge and were more satisfied with implementation supports than PaU participants. CoPs present a promising model for translating EBP knowledge and promoting practice change in children's mental health that requires further study.

  8. Regulation of the glv Operon in Bacillus subtilis: YfiA (GlvR) Is a Positive Regulator of the Operon That Is Repressed through CcpA and cre

    PubMed Central

    Yamamoto, Hiroki; Serizawa, Masakuni; Thompson, John; Sekiguchi, Junichi

    2001-01-01

    Maltose metabolism and the regulation of the glv operon of Bacillus subtilis, comprising three genes, glvA (6-phospho-α-glucosidase), yfiA (now designated glvR), and glvC (EIICB transport protein), were investigated. Maltose dissimilation was dependent primarily upon the glv operon, and insertional inactivation of either glvA, glvR, or glvC markedly inhibited growth on the disaccharide. A second system (MalL) contributed to a minor extent to maltose metabolism. Northern blotting revealed two transcripts corresponding to a monocistronic mRNA of glvA and a polycistronic mRNA of glvA-glvR-glvC. Primer extension analysis showed that both transcripts started at the same base (G) located 26 bp upstream of the 5′ end of glvA. When glvR was placed under control of the spac promoter, expression of the glv operon was dependent upon the presence of isopropyl-β-d-thiogalactopyranoside (IPTG). In regulatory studies, the promoter sequence of the glv operon was fused to lacZ and inserted into the amyE locus, and the resultant strain (AMGLV) was then transformed with a citrate-controlled glvR plasmid, pHYCM2VR. When cultured in Difco sporulation medium containing citrate, this transformant [AMGLV(pHYCM2VR)] expressed LacZ activity, but synthesis of LacZ was repressed by glucose. In an isogenic strain, [AMGLVCR(pHYCM2VR)], except for a mutation in the sequence of a catabolite-responsive element (cre), LacZ activity was expressed in the presence of citrate and glucose. Insertion of a citrate-controlled glvR plasmid at the amyE locus of ccpA+ and ccpA mutant organisms yielded strains AMCMVR and AMCMVRCC, respectively. In the presence of both glucose and citrate, AMCMVR failed to express the glv operon, whereas under the same conditions high-level expression of both mRNA transcripts was found in strain AMCMVRCC. Collectively, our findings suggest that GlvR (the product of the glvR gene) is a positive regulator of the glv operon and that glucose exerts its effect via catabolite repression requiring both CcpA and cre. PMID:11489864

  9. Long-range transcriptional control of an operon necessary for virulence-critical ESX-1 secretion in Mycobacterium tuberculosis.

    PubMed

    Hunt, Debbie M; Sweeney, Nathan P; Mori, Luisa; Whalan, Rachael H; Comas, Iñaki; Norman, Laura; Cortes, Teresa; Arnvig, Kristine B; Davis, Elaine O; Stapleton, Melanie R; Green, Jeffrey; Buxton, Roger S

    2012-05-01

    The ESX-1 secretion system of Mycobacterium tuberculosis has to be precisely regulated since the secreted proteins, although required for a successful virulent infection, are highly antigenic and their continued secretion would alert the immune system to the infection. The transcription of a five-gene operon containing espACD-Rv3613c-Rv3612c, which is required for ESX-1 secretion and is essential for virulence, was shown to be positively regulated by the EspR transcription factor. Thus, transcription from the start site, found to be located 67 bp upstream of espA, was dependent upon EspR enhancer-like sequences far upstream (between 884 and 1,004 bp), which we term the espA activating region (EAR). The EAR contains one of the known binding sites for EspR, providing the first in vivo evidence that transcriptional activation at the espA promoter occurs by EspR binding to the EAR and looping out DNA between this site and the promoter. Regulation of transcription of this operon thus takes place over long regions of the chromosome. This regulation may differ in some members of the M. tuberculosis complex, including Mycobacterium bovis, since deletions of the intergenic region have removed the upstream sequence containing the EAR, resulting in lowered espA expression. Consequent differences in expression of ESX-1 in these bacteria may contribute to their various pathologies and host ranges. The virulence-critical nature of this operon means that transcription factors controlling its expression are possible drug targets.

  10. A Structure-Based Mechanism for Arf1-Dependent Recruitment of Coatomer to Membranes

    PubMed Central

    Yu, Xinchao; Breitman, Marianna; Goldberg, Jonathan

    2012-01-01

    Summary Budding of COPI-coated vesicles from Golgi membranes requires an Arf-family G protein and the coatomer complex recruited from cytosol. Arf is also required with coatomer-related clathrin adaptor complexes to bud vesicles from the trans-Golgi network and endosomal compartments. To understand the structural basis for Arf-dependent recruitment of a vesicular coat to the membrane, we determined the structure of Arf1 bound to the γζ-COP subcomplex of coatomer. Structure-guided biochemical analysis reveals that a second Arf1-GTP molecule binds to βδ-COP at a site common to the γ- and β-COP subunits. The Arf1-binding sites on coatomer are spatially related to PtdIns4,5P2-binding sites on the endocytic AP2 complex, providing evidence that the orientation of membrane binding is general for this class of vesicular coat proteins. A bivalent GTP-dependent binding mode has implications for the dynamics of coatomer interaction with the Golgi and for the selection of cargo molecules. PMID:22304919

  11. Competing for space in an already crowded market: a mixed methods study of why an online community of practice (CoP) for alcohol harm reduction failed to generate interest amongst the group of public health professionals at which it was aimed.

    PubMed

    Ponsford, Ruth; Ford, Jennifer; Korjonen, Helena; Hughes, Emma; Keswani, Asha; Pliakas, Triantafyllos; Egan, Matt

    2017-07-21

    Improving mechanisms for knowledge translation (KT) and connecting decision-makers to each other and the information and evidence they consider relevant to their work remains a priority for public health. Virtual communities of practices (CoPs) potentially offer an affordable and flexible means of encouraging connection and sharing of evidence, information and learning among the public health community in ways that transgress traditional geographical, professional, institutional and time boundaries. The suitability of online CoPs in public health, however, has rarely been tested. This paper explores the reasons why particular online CoP for alcohol harm reduction hosted by the UK Health Forum failed to generate sufficient interest from the group of public health professionals at which it was aimed. The study utilises online web-metrics demonstrating a lack of online activity on the CoP. One hundred and twenty seven responses to an online questionnaire were used to explore whether the lack of activity could be explained by the target audience's existing information and evidence practices and needs. Qualitative interviews with 10 members describe in more detail the factors that shape and inhibit use of the virtual CoP by those at which it was targeted. Quantitative and qualitative data confirm that the target audience had an interest in the kind of information and evidence the CoP was set up to share and generate discussion about, but also that participants considered themselves to already have relatively good access to the information and evidence they needed to inform their work. Qualitative data revealed that the main barriers to using the CoP were a proliferation of information sources meaning that participants preferred to utilise trusted sources that were already established within their daily routines and a lack of time to engage with new online tools that required any significant commitment. Specialist online CoPs are competing for space in an already crowded market. A target audience that regards itself as busy and over-supplied is unlikely to commit to a new service without the assurance that the service will provide unique and valuable well-summarised information, which would reduce the need to spend time accessing competing resources.

  12. Use of communities of practice in business and health care sectors: a systematic review.

    PubMed

    Li, Linda C; Grimshaw, Jeremy M; Nielsen, Camilla; Judd, Maria; Coyte, Peter C; Graham, Ian D

    2009-05-17

    Since being identified as a concept for understanding knowledge sharing, management, and creation, communities of practice (CoPs) have become increasingly popular within the health sector. The CoP concept has been used in the business sector for over 20 years, but the use of CoPs in the health sector has been limited in comparison. First, we examined how CoPs were defined and used in these two sectors. Second, we evaluated the evidence of effectiveness on the health sector CoPs for improving the uptake of best practices and mentoring new practitioners. We conducted a search of electronic databases in the business, health, and education sectors, and a hand search of key journals for primary studies on CoP groups. Our research synthesis for the first objective focused on three areas: the authors' interpretations of the CoP concept, the key characteristics of CoP groups, and the common elements of CoP groups. To examine the evidence on the effectiveness of CoPs in the health sector, we identified articles that evaluated CoPs for improving health professional performance, health care organizational performance, professional mentoring, and/or patient outcome; and used experimental, quasi-experimental, or observational designs. The structure of CoP groups varied greatly, ranging from voluntary informal networks to work-supported formal education sessions, and from apprentice training to multidisciplinary, multi-site project teams. Four characteristics were identified from CoP groups: social interaction among members, knowledge sharing, knowledge creation, and identity building; however, these were not consistently present in all CoPs. There was also a lack of clarity in the responsibilities of CoP facilitators and how power dynamics should be handled within a CoP group. We did not find any paper in the health sector that met the eligibility criteria for the quantitative analysis, and so the effectiveness of CoP in this sector remained unclear. There is no dominant trend in how the CoP concept is operationalized in the business and health sectors; hence, it is challenging to define the parameters of CoP groups. This may be one of the reasons for the lack of studies on the effectiveness of CoPs in the health sector. In order to improve the usefulness of the CoP concept in the development of groups and teams, further research will be needed to clarify the extent to which the four characteristics of CoPs are present in the mature and emergent groups, the expectations of facilitators and other participants, and the power relationship within CoPs.

  13. SMA Diagnosis: Detection of SMN1 Deletion with Real-Time mCOP-PCR System Using Fresh Blood DNA.

    PubMed

    Niba, Emma Tabe Eko; Ar Rochmah, Mawaddah; Harahap, Nur Imma Fatimah; Awano, Hiroyuki; Morioka, Ichiro; Iijima, Kazumoto; Saito, Toshio; Saito, Kayoko; Takeuchi, Atsuko; Lai, Poh San; Bouike, Yoshihiro; Nishio, Hisahide; Shinohara, Masakazu

    2017-12-18

    Spinal muscular atrophy (SMA) is one of the most common autosomal recessive disorders. The symptoms are caused by defects of lower motor neurons in the spinal cord. More than 95% of SMA patients are homozygous for survival motor neuron 1 (SMN1) deletion. We previously developed a screening system for SMN1 deletion based on a modified competitive oligonucleotide priming-PCR (mCOP-PCR) technique using dried blood spot (DBS) on filter paper. This system is convenient for mass screening in the large population and/or first-tier diagnostic method of the patients in the remote areas. However, this system was still time-consuming and effort-taking, because it required pre-amplification procedure to avoid non-specific amplification and gel-electrophoresis to detect the presence or absence of SMN1 deletion. When the fresh blood samples are used instead of DBS, or when the gel-electrophoresis is replaced by real-time PCR, we may have a simpler and more rapid diagnostic method for SMA. To establish a simpler and more rapid diagnostic method of SMN1 deletion using fresh blood DNA. DNA samples extracted from fresh blood and stored at 4 ℃ for 1 month. The samples were assayed using a real-time mCOP-PCR system without pre-amplification procedures. DNA samples had already been genotyped by PCR-restriction fragment length polymorphism (PCR-RFLP), showing the presence or absence of SMN1 exon 7. The DNA samples were directly subjected to the mCOP-PCR step. The amplification of mCOP-PCR was monitored in a real-time PCR apparatus. The genotyping results of the real-time mCOP-PCR system using fresh blood DNA were completely matched with those of PCR-RFLP. In this real-time mCOP-PCR system using fresh blood-DNA, it took only four hours from extraction of DNA to detection of the presence or absence of SMN1 deletion, while it took more than 12 hours in PCR-RFLP. Our real-time mCOP-PCR system using fresh blood DNA was rapid and accurate, suggesting it may be useful for the first-tier diagnostic method of SMA.

  14. McMYB10 Modulates the Expression of a Ubiquitin Ligase, McCOP1 During Leaf Coloration in Crabapple

    PubMed Central

    Li, Ke-Ting; Zhang, Jie; Kang, Yan-Hui; Chen, Meng-Chen; Song, Ting-Ting; Geng, Hui; Tian, Ji; Yao, Yun-Cong

    2018-01-01

    In higher plants, anthocyanins are protective secondary metabolites, which contribute to the color of leaves, stems, flowers, and fruits, and have been found to have an antioxidant role in human health. In this study, we determined the expression of McMYB10 and its specific E3 ubiquitin ligase, McCOP1, in crabapple leaves during the course of a day and in five leaf development stages. Interestingly, the results showed that the transcription level of McCOP1 genes was higher in daylight than at night, and the transcripts of McMYB10 presented a positive correlation with the transcription of McCOP1-1 and McCOP1-2 and anthocyanin accumulation in a crabapple cultivar with red-colored leaves. Several MYB transcription factor (TF) binding sites of the MYBCORE type were found in the McCOP1-1 and McCOP1-2 promoters, and we deduced that there may be a relationship between McMYB10 and McCOP1-1 and McCOP1-2 at the transcriptional level. Yeast one hybrid (Y1H) and electrophoretic mobility shift assays (EMSA) demonstrated that the McMYB10 TF binds specifically to the promoter of McCOP1-1 and McCOP1-2. Furthermore, increased levels of McMYB10 promoted anthocyanin biosynthesis and the expression level of McCOP1-1 and McCOP1-2 in crabapple leaves during continuous light treatments, and overexpression or silencing of McMYB10 in crabapple leaves and apple fruits also result in an increase or decrease, respectively, in the expression of McCOP1-1 and McCOP1-2 and in anthocyanin biosynthesis. Our results reveal a new self-regulation mechanism in where McMYB10 modulates its own expression by activating McCOP1-1 and McCOP1-2 expression to promote ubiquitination of the McMYB10 protein by McCOP1. PMID:29915606

  15. Transcriptional Regulator LsrB of Sinorhizobium meliloti Positively Regulates the Expression of Genes Involved in Lipopolysaccharide Biosynthesis

    PubMed Central

    Tang, Guirong; Wang, Ying

    2014-01-01

    Rhizobia induce nitrogen-fixing nodules on host legumes, which is important in agriculture and ecology. Lipopolysaccharide (LPS) produced by rhizobia is required for infection or bacteroid survival in host cells. Genes required for LPS biosynthesis have been identified in several Rhizobium species. However, the regulation of their expression is not well understood. Here, Sinorhizobium meliloti LsrB, a member of the LysR family of transcriptional regulators, was found to be involved in LPS biosynthesis by positively regulating the expression of the lrp3-lpsCDE operon. An lsrB in-frame deletion mutant displayed growth deficiency, sensitivity to the detergent sodium dodecyl sulfate, and acidic pH compared to the parent strain. This mutant produced slightly less LPS due to lower expression of the lrp3 operon. Analysis of the transcriptional start sites of the lrp3 and lpsCDE gene suggested that they constitute one operon. The expression of lsrB was positively autoregulated. The promoter region of lrp3 was specifically precipitated by anti-LsrB antibodies in vivo. The promoter DNA fragment containing TN11A motifs was bound by the purified LsrB protein in vitro. These new findings suggest that S. meliloti LsrB is associated with LPS biosynthesis, which is required for symbiotic nitrogen fixation on some ecotypes of alfalfa plants. PMID:24951786

  16. Triple effect absorption cycles

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

    Erickson, D.C.; Potnis, S.V.; Tang, J.

    1996-12-31

    Triple effect absorption chillers can achieve 50% COP improvement over double-effect systems. However, to translate this potential into cost-effective hardware, the most promising embodiments must be identified. In this study, 12 generic triple effect cycles and 76 possible hermetic loop arrangements of those 12 generic cycles were identified. The generic triple effect cycles were screened based on their pressure and solubility field requirements, generic COPs, risk involved in the component design, and number of components in a high corrosive environment. This screening identified four promising arrangements: Alkitrate Topping cycle, Pressure Staged Envelope cycle, High Pressure Overlap cycle, and Dual Loopmore » cycle. All of these arrangements have a very high COP ({approximately} 1.8), however the development risk and cost involved is different for each arrangement. Therefore, the selection of a particular arrangement will depend upon the specific situation under consideration.« less

  17. The Pea light-independent photomorphogenesis1 Mutant Results from Partial Duplication of COP1 Generating an Internal Promoter and Producing Two Distinct Transcripts

    PubMed Central

    Sullivan, James A.; Gray, John C.

    2000-01-01

    The pea lip1 (light-independent photomorphogenesis1) mutant shows many of the characteristics of light-grown development when grown in continuous darkness. To investigate the identity of LIP1, cDNAs encoding the pea homolog of COP1, a repressor of photomorphogenesis identified in Arabidopsis, were isolated from wild-type and lip1 pea seedlings. lip1 seedlings contained a wild-type COP1 transcript as well as a larger COP1′ transcript that contained an internal in-frame duplication of 894 bp. The COP1′ transcript segregated with the lip1 phenotype in F2 seedlings and could be translated in vitro to produce a protein of ∼100 kD. The COP1 gene in lip1 peas contained a 7.5-kb duplication, consisting of exons 1 to 7 of the wild-type sequence, located 2.5 kb upstream of a region of genomic DNA identical to the wild-type COP1 DNA sequence. Transcription and splicing of the mutant COP1 gene was predicted to produce the COP1′ transcript, whereas transcription from an internal promoter in the 2.5-kb region of DNA located between the duplicated regions of COP1 would produce the wild-type COP1 transcript. The presence of small quantities of wild-type COP1 transcripts may reduce the severity of the phenotype produced by the mutated COP1′ protein. The genomic DNA sequences of the COP1 gene from wild-type and lip1 peas and the cDNA sequences of COP1 and COP1′ transcripts have been submitted to the EMBL database under the EMBL accession numbers AJ276591, AJ276592, AJ289773, and AJ289774, respectively. PMID:11041887

  18. Use of communities of practice in business and health care sectors: A systematic review

    PubMed Central

    Li, Linda C; Grimshaw, Jeremy M; Nielsen, Camilla; Judd, Maria; Coyte, Peter C; Graham, Ian D

    2009-01-01

    Background Since being identified as a concept for understanding knowledge sharing, management, and creation, communities of practice (CoPs) have become increasingly popular within the health sector. The CoP concept has been used in the business sector for over 20 years, but the use of CoPs in the health sector has been limited in comparison. Objectives First, we examined how CoPs were defined and used in these two sectors. Second, we evaluated the evidence of effectiveness on the health sector CoPs for improving the uptake of best practices and mentoring new practitioners. Methods We conducted a search of electronic databases in the business, health, and education sectors, and a hand search of key journals for primary studies on CoP groups. Our research synthesis for the first objective focused on three areas: the authors' interpretations of the CoP concept, the key characteristics of CoP groups, and the common elements of CoP groups. To examine the evidence on the effectiveness of CoPs in the health sector, we identified articles that evaluated CoPs for improving health professional performance, health care organizational performance, professional mentoring, and/or patient outcome; and used experimental, quasi-experimental, or observational designs. Results The structure of CoP groups varied greatly, ranging from voluntary informal networks to work-supported formal education sessions, and from apprentice training to multidisciplinary, multi-site project teams. Four characteristics were identified from CoP groups: social interaction among members, knowledge sharing, knowledge creation, and identity building; however, these were not consistently present in all CoPs. There was also a lack of clarity in the responsibilities of CoP facilitators and how power dynamics should be handled within a CoP group. We did not find any paper in the health sector that met the eligibility criteria for the quantitative analysis, and so the effectiveness of CoP in this sector remained unclear. Conclusion There is no dominant trend in how the CoP concept is operationalized in the business and health sectors; hence, it is challenging to define the parameters of CoP groups. This may be one of the reasons for the lack of studies on the effectiveness of CoPs in the health sector. In order to improve the usefulness of the CoP concept in the development of groups and teams, further research will be needed to clarify the extent to which the four characteristics of CoPs are present in the mature and emergent groups, the expectations of facilitators and other participants, and the power relationship within CoPs. PMID:19445723

  19. 20 CFR 10.216 - How is the pay rate for COP calculated?

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 20 Employees' Benefits 1 2014-04-01 2012-04-01 true How is the pay rate for COP calculated? 10.216... AMENDED Continuation of Pay Calculation of Cop § 10.216 How is the pay rate for COP calculated? The employer shall calculate COP using the period of time and the weekly pay rate. (a) The pay rate for COP...

  20. 20 CFR 10.216 - How is the pay rate for COP calculated?

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 20 Employees' Benefits 1 2013-04-01 2012-04-01 true How is the pay rate for COP calculated? 10.216... AMENDED Continuation of Pay Calculation of Cop § 10.216 How is the pay rate for COP calculated? The employer shall calculate COP using the period of time and the weekly pay rate. (a) The pay rate for COP...

  1. The nucleoid-associated protein Fis directly modulates the synthesis of cellulose, an essential component of pellicle-biofilms in the phytopathogenic bacterium Dickeya dadantii.

    PubMed

    Prigent-Combaret, Claire; Zghidi-Abouzid, Ouafa; Effantin, Géraldine; Lejeune, Philippe; Reverchon, Sylvie; Nasser, William

    2012-10-01

    Bacteria use biofilm structures to colonize surfaces and to survive in hostile conditions, and numerous bacteria produce cellulose as a biofilm matrix polymer. Hence, expression of the bcs operon, responsible for cellulose biosynthesis, must be finely regulated in order to allow bacteria to adopt the proper surface-associated behaviours. Here we show that in the phytopathogenic bacterium, Dickeya dadantii, production of cellulose is required for pellicle-biofilm formation and resistance to chlorine treatments. Expression of the bcs operon is growth phase-regulated and is stimulated in biofilms. Furthermore, we unexpectedly found that the nucleoid-associated protein and global regulator of virulence functions, Fis, directly represses bcs operon expression by interacting with an operator that is absent from the bcs operon of animal pathogenic bacteria and the plant pathogenic bacterium Pectobacterium. Moreover, production of cellulose enhances plant surface colonization by D. dadantii. Overall, these data suggest that cellulose production and biofilm formation may be important factors for surface colonization by D. dadantii and its subsequent survival in hostile environments. This report also presents a new example of how bacteria can modulate the action of a global regulator to co-ordinate basic metabolism, virulence and modifications of lifestyle. © 2012 Blackwell Publishing Ltd.

  2. Awakening sleeping beauty: production of propionic acid in Escherichia coli through the sbm operon requires the activity of a methylmalonyl-CoA epimerase.

    PubMed

    Gonzalez-Garcia, Ricardo Axayacatl; McCubbin, Tim; Wille, Annalena; Plan, Manuel; Nielsen, Lars Keld; Marcellin, Esteban

    2017-07-17

    Propionic acid is used primarily as a food preservative with smaller applications as a chemical building block for the production of many products including fabrics, cosmetics, drugs, and plastics. Biological production using propionibacteria would be competitive against chemical production through hydrocarboxylation of ethylene if native producers could be engineered to reach near-theoretical yield and good productivity. Unfortunately, engineering propionibacteria has proven very challenging. It has been suggested that activation of the sleeping beauty operon in Escherichia coli is sufficient to achieve propionic acid production. Optimising E. coli production should be much easier than engineering propionibacteria if tolerance issues can be addressed. Propionic acid is produced in E. coli via the sleeping beauty mutase operon under anaerobic conditions in rich medium via amino acid degradation. We observed that the sbm operon enhances amino acids degradation to propionic acid and allows E. coli to degrade isoleucine. However, we show here that the operon lacks an epimerase reaction that enables propionic acid production in minimal medium containing glucose as the sole carbon source. Production from glucose can be restored by engineering the system with a methylmalonyl-CoA epimerase from Propionibacterium acidipropionici (0.23 ± 0.02 mM). 1-Propanol production was also detected from the promiscuous activity of the native alcohol dehydrogenase (AdhE). We also show that aerobic conditions are favourable for propionic acid production. Finally, we increase titre 65 times using a combination of promoter engineering and process optimisation. The native sbm operon encodes an incomplete pathway. Production of propionic acid from glucose as sole carbon source is possible when the pathway is complemented with a methylmalonyl-CoA epimerase. Although propionic acid via the restored succinate dissimilation pathway is considered a fermentative process, the engineered pathway was shown to be functional under anaerobic and aerobic conditions.

  3. Insect peptide CopA3-induced protein degradation of p27Kip1 stimulates proliferation and protects neuronal cells from apoptosis

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

    Nam, Seung Taek; Kim, Dae Hong; Lee, Min Bum

    2013-07-19

    Highlights: •CopA3 peptide isolated from the Korean dung beetle has antimicrobial activity. •Our study reported that CopA3 has anticancer and immunosuppressive effects. •We here demonstrated that CopA3 has neurotropic and neuroprotective effects. •CopA3 degrades p27Kip1 protein and this mediates effects of CopA3 on neuronal cells. -- Abstract: We recently demonstrated that the antibacterial peptide, CopA3 (a D-type disulfide dimer peptide, LLCIALRKK), inhibits LPS-induced macrophage activation and also has anticancer activity in leukemia cells. Here, we examined whether CopA3 could affect neuronal cell proliferation. We found that CopA3 time-dependently increased cell proliferation by up to 31 ± 2% in human neuroblastomamore » SH-SY5Y cells, and up to 29 ± 2% in neural stem cells isolated from neonatal mouse brains. In both cell types, CopA3 also significantly inhibited the apoptosis and viability losses caused by 6-hydroxy dopamine (a Parkinson disease-mimicking agent) and okadaic acid (an Alzheimer’s disease-mimicking agent). Immunoblotting revealed that the p27Kip1 protein (a negative regulator of cell cycle progression) was markedly degraded in CopA3-treated SH-SY5Y cells. Conversely, an adenovirus expressing p27Kip1 significantly inhibited the antiapoptotic effects of CopA3 against 6-hydroxy dopamine- and okadaic acid-induced apoptosis, and decreased the neurotropic effects of CopA3. These results collectively suggest that CopA3-mediated protein degradation of p27Kip1 may be the main mechanism through which CopA3 exerts neuroprotective and neurotropic effects.« less

  4. Depletion of ε-COP in the COPI Vesicular Coat Reduces Cleistothecium Production in Aspergillus nidulans.

    PubMed

    Kang, Eun-Hye; Song, Eun-Jung; Kook, Jun Ho; Lee, Hwan-Hee; Jeong, Bo-Ri; Park, Hee-Moon

    2015-03-01

    We have previously isolated ε-COP, the α-COP interactor in COPI of Aspergillus nidulans, by yeast two-hybrid screening. To understand the function of ε-COP, the aneA (+) gene for ε-COP/AneA was deleted by homologous recombination using a gene-specific disruption cassette. Deletion of the ε-COP gene showed no detectable changes in vegetative growth or asexual development, but resulted in decrease in the production of the fruiting body, cleistothecium, under conditions favorable for sexual development. Unlike in the budding yeast Saccharomyces cerevisiae, in A. nidulans, over-expression of ε-COP did not rescue the thermo-sensitive growth defect of the α-COP mutant at 42℃. Together, these data show that ε-COP is not essential for viability, but it plays a role in fruiting body formation in A. nidulans.

  5. Optimization for ultrasonic-microwave synergistic extraction of polysaccharides from Cornus officinalis and characterization of polysaccharides.

    PubMed

    Yin, Xiulian; You, Qinghong; Jiang, Zhonghai; Zhou, Xinghai

    2016-02-01

    Ultrasonic-microwave synergistic extraction (UMSE) of polysaccharides from Cornus officinalis was optimized by response surface methodology (RSM). The effect of four different factors on the yield of C. officinalis polysaccharides (COP) was studied. RSM results showed that the optimal conditions were extraction time of 31.49823 min, microwave power of 99.39769 W, and water-to-raw material ratio of 28.16273. The COP yield was 11.38±0.31% using the modified optimal conditions, which was consistent with the value predicted by the model. The crude COP was purified by DEAE-Cellulose 52 chromatography and Sephadex G-100 chromatography. Five fractions, namely, crude COP, COP-1, COP-2, COP-3, and COP-4, were obtained. Monosaccharide composition analysis revealed that the COP was composed of glucose, arabinose, fucose, xylose, mannose, and rhamnose. Preliminary structural characterizations of COP were conducted by scanning electron microscopy and Fourier transform infrared spectroscopy. Copyright © 2015 Elsevier B.V. All rights reserved.

  6. The Ubiquitin Ligase COP1 Promotes Glioma Cell Proliferation by Preferentially Downregulating Tumor Suppressor p53.

    PubMed

    Zou, Shenshan; Zhu, Yufu; Wang, Bin; Qian, Fengyuan; Zhang, Xiang; Wang, Lei; Fu, Chunling; Bao, Hanmo; Xie, Manyi; Gao, Shangfeng; Yu, Rutong; Shi, Hengliang

    2017-09-01

    Human glioma causes substantial morbidity and mortality worldwide. However, the molecular mechanisms underlying glioma progression are still largely unknown. COP1 (constitutively photomorphogenic 1), an E3 ubiquitin ligase, is important in cell survival, development, cell growth, and cancer biology by regulating different substrates. As is well known, both tumor suppressor p53 and oncogenic protein c-JUN could be ubiquitinated and degraded by ubiquitin ligase COP1, which may be the reason that COP1 serves as an oncogene or a tumor suppressor in different cancer types. Up to now, the possible role of COP1 in human glioma is still unclear. In the present study, we found that the expression of COP1 was upregulated in human glioma tissues. The role of COP1 in glioma cell proliferation was investigated using COP1 loss- and gain-of-function. The results showed that downregulation of COP1 by short hairpin RNA (shRNA) inhibited glioma cell proliferation, while overexpression of COP1 significantly promoted it. Furthermore, we demonstrated that COP1 only interacted with and regulated p53, but not c-JUN. Taken together, these results indicate that COP1 may play a role in promoting glioma cell proliferation by interacting with and downregulating tumor suppressor p53 rather than oncogenic protein c-JUN.

  7. Comparison of the overlapping frd and ampC operons of Escherichia coli with the corresponding DNA sequences in other gram-negative bacteria.

    PubMed

    Bergström, S; Lindberg, F P; Olsson, O; Normark, S

    1983-09-01

    Specific DNA probes from Escherichia coli K-12 were used to analyze the sequence divergence of the frd and ampC operons in various species of gram-negative bacteria. These operons code for the fumarate reductase complex and the chromosomal beta-lactamase, respectively. We demonstrate that the two operons show the same general pattern of divergence, although the frd operon is considerably more conserved than is the ampC operon. The major exception is Salmonella typhimurium LT2, which shows a strong homology to the E. coli frd probe but none to the E. coli ampC probe. The operons from Citrobacter freundii and Shigella sonnei were cloned and characterized by physical mapping, Southern hybridization, and protein synthesis in minicells. In S. sonnei, as in E. coli K-12, the frd and ampC operons overlap (T. Grundström and B. Jaurin, Proc. Natl. Acad. Sci. U.S.A. 79:1111-1115, 1982). Only minor discrepancies between the two operons were found over the entire frd-ampC region. In C. freundii, the ampC and frd operons do not overlap, being separated by about 1,100 base pairs. Presumably the inducible property of the C. freundii chromosomal beta-lactamase is encoded by this 1,100-base-pair DNA segment.

  8. Interplay of Noisy Gene Expression and Dynamics Explains Patterns of Bacterial Operon Organization

    NASA Astrophysics Data System (ADS)

    Igoshin, Oleg

    2011-03-01

    Bacterial chromosomes are organized into operons -- sets of genes co-transcribed into polycistronic messenger RNA. Hypotheses explaining the emergence and maintenance of operons include proportional co-regulation, horizontal transfer of intact ``selfish'' operons, emergence via gene duplication, and co-production of physically interacting proteins to speed their association. We hypothesized an alternative: operons can reduce or increase intrinsic gene expression noise in a manner dependent on the post-translational interactions, thereby resulting in selection for or against operons in depending on the network architecture. We devised five classes of two-gene network modules and show that the effects of operons on intrinsic noise depend on class membership. Two classes exhibit decreased noise with co-transcription, two others reveal increased noise, and the remaining one does not show a significant difference. To test our modeling predictions we employed bioinformatic analysis to determine the relationship gene expression noise and operon organization. The results confirm the overrepresentation of noise-minimizing operon architectures and provide evidence against other hypotheses. Our results thereby suggest a central role for gene expression noise in selecting for or maintaining operons in bacterial chromosomes. This demonstrates how post-translational network dynamics may provide selective pressure for organizing bacterial chromosomes, and has practical consequences for designing synthetic gene networks. This work is supported by National Institutes of Health grant 1R01GM096189-01.

  9. Operon Conservation and the Evolution of trans-Splicing in the Phylum Nematoda

    PubMed Central

    Guiliano, David B; Blaxter, Mark L

    2006-01-01

    The nematode Caenorhabditis elegans is unique among model animals in that many of its genes are cotranscribed as polycistronic pre-mRNAs from operons. The mechanism by which these operonic transcripts are resolved into mature mRNAs includes trans-splicing to a family of SL2-like spliced leader exons. SL2-like spliced leaders are distinct from SL1, the major spliced leader in C. elegans and other nematode species. We surveyed five additional nematode species, representing three of the five major clades of the phylum Nematoda, for the presence of operons and the use of trans-spliced leaders in resolution of polycistronic pre-mRNAs. Conserved operons were found in Pristionchus pacificus, Nippostrongylus brasiliensis, Strongyloides ratti, Brugia malayi, and Ascaris suum. In nematodes closely related to the rhabditine C. elegans, a related family of SL2-like spliced leaders is used for operonic transcript resolution. However, in the tylenchine S. ratti operonic transcripts are resolved using a family of spliced leaders related to SL1. Non-operonic genes in S. ratti may also receive these SL1 variants. In the spirurine nematodes B. malayi and A. suum operonic transcripts are resolved using SL1. Mapping these phenotypes onto the robust molecular phylogeny for the Nematoda suggests that operons evolved before SL2-like spliced leaders, which are an evolutionary invention of the rhabditine lineage. PMID:17121468

  10. Glycopeptide Resistance vanA Operons in Paenibacillus Strains Isolated from Soil

    PubMed Central

    Guardabassi, Luca; Perichon, Bruno; van Heijenoort, Jean; Blanot, Didier; Courvalin, Patrice

    2005-01-01

    The sequence and gene organization of the van operons in vancomycin (MIC of >256 μg/ml)- and teicoplanin (MIC of ≥32 μg/ml)-resistant Paenibacillus thiaminolyticus PT-2B1 and Paenibacillus apiarius PA-B2B isolated from soil were determined. Both operons had regulatory (vanR and vanS), resistance (vanH, vanA, and vanX), and accessory (vanY, vanZ, and vanW) genes homologous to the corresponding genes in enterococcal vanA and vanB operons. The vanAPT operon in P. thiaminolyticus PT-2B1 had the same gene organization as that of vanA operons whereas vanAPA in P. apiarius PA-B2B resembled vanB operons due to the presence of vanW upstream from the vanHAX cluster but was closer to vanA operons in sequence. Reference P. apiarius strains NRRL B-4299 and NRRL B-4188 were found to harbor operons indistinguishable from vanAPA by PCR mapping, restriction fragment length polymorphism, and partial sequencing, suggesting that this operon was species specific. As in enterococci, resistance was inducible by glycopeptides and associated with the synthesis of pentadepsipeptide peptidoglycan precursors ending in d-Ala-d-Lac, as demonstrated by d,d-dipeptidase activities, high-pressure liquid chromatography, and mass spectrometry. The precursors differed from those in enterococci by the presence of diaminopimelic acid instead of lysine in the peptide chain. Altogether, the results are compatible with the notion that van operons in soil Paenibacillus strains and in enterococci have evolved from a common ancestor. PMID:16189102

  11. Glycopeptide resistance vanA operons in Paenibacillus strains isolated from soil.

    PubMed

    Guardabassi, Luca; Perichon, Bruno; van Heijenoort, Jean; Blanot, Didier; Courvalin, Patrice

    2005-10-01

    The sequence and gene organization of the van operons in vancomycin (MIC of >256 microg/ml)- and teicoplanin (MIC of > or =32 microg/ml)-resistant Paenibacillus thiaminolyticus PT-2B1 and Paenibacillus apiarius PA-B2B isolated from soil were determined. Both operons had regulatory (vanR and vanS), resistance (vanH, vanA, and vanX), and accessory (vanY, vanZ, and vanW) genes homologous to the corresponding genes in enterococcal vanA and vanB operons. The vanA(PT) operon in P. thiaminolyticus PT-2B1 had the same gene organization as that of vanA operons whereas vanA(PA) in P. apiarius PA-B2B resembled vanB operons due to the presence of vanW upstream from the vanHAX cluster but was closer to vanA operons in sequence. Reference P. apiarius strains NRRL B-4299 and NRRL B-4188 were found to harbor operons indistinguishable from vanA(PA) by PCR mapping, restriction fragment length polymorphism, and partial sequencing, suggesting that this operon was species specific. As in enterococci, resistance was inducible by glycopeptides and associated with the synthesis of pentadepsipeptide peptidoglycan precursors ending in D-Ala-D-Lac, as demonstrated by D,D-dipeptidase activities, high-pressure liquid chromatography, and mass spectrometry. The precursors differed from those in enterococci by the presence of diaminopimelic acid instead of lysine in the peptide chain. Altogether, the results are compatible with the notion that van operons in soil Paenibacillus strains and in enterococci have evolved from a common ancestor.

  12. Insights into arsenic multi-operons expression and resistance mechanisms in Rhodopseudomonas palustris CGA009

    PubMed Central

    Zhao, Chungui; Zhang, Yi; Chan, Zhuhua; Chen, Shicheng; Yang, Suping

    2015-01-01

    Arsenic (As) is widespread in the environment and causes numerous health problems. Rhodopseudomonas palustris has been regarded as a good model organism for studying arsenic detoxification since it was first demonstrated to methylate environmental arsenic by conversion to soluble or gaseous methylated species. However, the detailed arsenic resistance mechanisms remain unknown though there are at least three arsenic-resistance operons (ars1, ars2, and ars3) in R. palustris. In this study, we investigated how arsenic multi-operons contributed to arsenic detoxification in R. palustris. The expression of ars2 or ars3 operons increased with increasing environmental arsenite (As(III)) concentrations (up to 1.0 mM) while transcript of ars1 operon was not detected in the middle log-phase (55 h). ars2 operon was actively expressed even at the low concentration of As(III) (0.01 μM), whereas the ars3 operon was expressed at 1.0 μM of As(III), indicating that there was a differential regulation mechanism for the three arsenic operons. Furthermore, ars2 and ars3 operons were maximally transcribed in the early log-phase where ars2 operon was 5.4-fold higher than that of ars3 operon. A low level of ars1 transcript was only detected at 43 h (early log-phase). Arsenic speciation analysis demonstrated that R. palustris could reduce As(V) to As(III). Collectively, strain CGA009 detoxified arsenic by using arsenic reduction and methylating arsenic mechanism, while the latter might occur with the presence of higher concentrations of arsenic. PMID:26441915

  13. Identification and characterization of the nickel uptake system for urease biogenesis in Streptococcus salivarius 57.I.

    PubMed

    Chen, Yi-Ywan M; Burne, Robert A

    2003-12-01

    Ureases are multisubunit enzymes requiring Ni(2+) for activity. The low pH-inducible urease gene cluster in Streptococcus salivarius 57.I is organized as an operon, beginning with ureI, followed by ureABC (structural genes), and ureEFGD (accessory genes). Urease biogenesis also requires a high-affinity Ni(2+) uptake system. By searching the partial genome sequence of a closely related organism, Streptococcus thermophilus LMG18311, three open reading frame (ORFs) homologous to those encoding proteins involved in cobalamin biosynthesis and cobalt transport (cbiMQO) were identified immediately 3' to the ure operon. To determine whether these genes were involved in urease biogenesis by catalyzing Ni(2+) uptake in S. salivarius, regions 3' to ureD were amplified by PCRs from S. salivarius by using primers identical to the S. thermophilus sequences. Sequence analysis of the products revealed three ORFs. Reverse transcriptase PCR was used to demonstrate that the ORFs are transcribed as part of the ure operon. Insertional inactivation of ORF1 with a polar kanamycin marker completely abolished urease activity and the ability to accumulate (63)Ni(2+) during growth. Supplementation of the growth medium with NiCl(2) at concentrations as low as 2.5 micro M partially restored urease activity in the mutant. Both wild-type and mutant strains showed enhanced urease activity when exogenous Ni(2+) was provided at neutral pH. Enhancement of urease activity by adding nickel was regulated at the posttranslational level. Thus, ORF1, ORF2, and ORF3 are part of the ure operon, and these genes, designated ureM, ureQ, and ureO, respectively, likely encode a Ni(2+)-specific ATP-binding cassette transporter.

  14. Getting to Uptake: Do Communities of Practice Support the Implementation of Evidence-Based Practice?

    PubMed Central

    Barwick, Melanie A.; Peters, Julia; Boydell, Katherine

    2009-01-01

    Introduction Practitioners are increasingly encouraged to adopt evidence-based practices (EBP) leading to a need for new knowledge translation strategies to support implementation and practice change. This study examined the benefits of a community of practice in the context of Ontario’s children’s mental health sector where organizations are mandated to adopt a standardized outcome measure to monitor client response to treatment. Method Readiness for change, practice change, content knowledge, and satisfaction with and use of implementation supports were examined among practitioners newly trained on the measure who were randomly assigned to a community of practice (CoP) or a practice as usual (PaU) group. CoP practitioners attended 6 sessions over 12 months; PaU practitioners had access to usual implementation supports. Results Groups did not differ on readiness for change or reported practice change, although CoP participants demonstrated greater use of the tool in practice, better content knowledge and were more satisfied with implementation supports than PaU participants. Conclusion CoPs present a promising model for translating EBP knowledge and promoting practice change in children’s mental health that requires further study. PMID:19270845

  15. THE EFFECTS OF DIFFERENT TRUNK INCLINATIONS ON BILATERAL TRUNK MUSCULAR ACTIVITIES, CENTRE OF PRESSURE AND FORCE EXERTIONS IN STATIC PUSHING POSTURES.

    PubMed

    Sanjaya, Kadek Heri; Lee, Soomin; Sriwarno, Andar Bagus; Shimomura, Yoshihito; Katsuura, Tetsuo

    2014-06-01

    In order to reconcile contradictory results from previous studies on manual pushing, a study was conducted to examine the effect of trunk inclination on muscular activities, centre of pressure (COP) and force exertion during static pushing. Ten subjects pushed at 0 degrees, 15 degrees, 30 degrees, and 45 degrees body inclinations in parallel and staggered feet stances. Wall and ground force plates measured pushing force, wall COP, vertical ground reaction force (GRF) and ground COP. Electromyogram data were recorded at 10 trunk muscle sites. Pushing force was found to increase with body inclination. GRF peaked at 15 degrees and reached its lowest level at the 45 degrees inclination. The lowest wall force plate standard deviation of COP displacement was found at the 30 degrees inclination. The lowest low back muscular activity was found at the 15 degrees and 30 degrees inclinations. Based on force exertion, muscular load, and stability, the 30 degrees body inclination was found to be the best posture for static pushing. This study also showed asymmetry in muscular activity and force exertion which has been received less attention in manual pushing studies. These findings will require further study.

  16. Study of Staphylococcus aureus N315 Pathogenic Genes by Text Mining and Enrichment Analysis of Pathways and Operons.

    PubMed

    Yang, Chun-Feng; Gou, Wei-Hui; Dai, Xin-Lun; Li, Yu-Mei

    2018-06-01

    Staphylococcus aureus (S. aureus) is a versatile pathogen found in many environments and can cause nosocomial infections in the community and hospitals. S. aureus infection is an increasingly serious threat to global public health that requires action across many government bodies, medical and health sectors, and scientific research institutions. In the present study, S. aureus N315 genes that have been shown in the literature to be pathogenic were extracted using a bibliometric method for functional enrichment analysis of pathways and operons to statistically discover novel pathogenic genes associated with S. aureus N315. A total of 383 pathogenic genes were mined from the literature using bibliometrics, and subsequently a few new pathogenic genes of S. aureus N315 were identified by functional enrichment analysis of pathways and operons. The discovery of these novel S. aureus N315 pathogenic genes is of great significance to treat S. aureus induced diseases and identify potential diagnostic markers, thus providing theoretical fundamentals for epidemiological prevention.

  17. Bistable Behavior of the Lac Operon in E. Coli When Induced with a Mixture of Lactose and TMG

    PubMed Central

    Díaz-Hernández, Orlando; Santillán, Moisés

    2010-01-01

    In this work we investigate multistability in the lac operon of Escherichia coli when it is induced by a mixture of lactose and the non-metabolizable thiomethyl galactoside (TMG). In accordance with previously published experimental results and computer simulations, our simulations predict that: (1) when the system is induced by TMG, the system shows a discernible bistable behavior while, (2) when the system is induced by lactose, bistability does not disappear but excessively high concentrations of lactose would be required to observe it. Finally, our simulation results predict that when a mixture of lactose and TMG is used, the bistability region in the extracellular glucose concentration vs. extracellular lactose concentration parameter space changes in such a way that the model predictions regarding bistability could be tested experimentally. These experiments could help to solve a recent controversy regarding the existence of bistability in the lac operon under natural conditions. PMID:21423364

  18. Regulation of the cnr Cobalt and Nickel Resistance Determinant from Ralstonia sp. Strain CH34†

    PubMed Central

    Grass, Gregor; Große, Cornelia; Nies, Dietrich H.

    2000-01-01

    Ralstonia sp. strain CH34 is resistant to nickel and cobalt cations. Resistance is mediated by the cnr determinant located on plasmid pMOL28. The cnr genes are organized in two clusters, cnrYXH and cnrCBA. As revealed by reverse transcriptase PCR and primer extension, transcription from these operons is initiated from promoters located upstream of the cnrY and cnrC genes. These two promoters exhibit conserved sequences at the −10 (CCGTATA) and −35 (CRAGGGGRAG) regions. The CnrH gene product, which is required for expression of both operons, is a sigma factor belonging to the sigma L family, whose activity seems to be governed by the membrane-bound CnrY and CnrX gene products in response to Ni2+. Half-maximal activation from the cnrCBA operon was determined by using appropriate lacZ gene fusions and was shown to occur at an Ni2+ concentration of about 50 μM. PMID:10671463

  19. Transcription coactivator Arabidopsis ANGUSTIFOLIA3 modulates anthocyanin accumulation and light-induced root elongation through transrepression of Constitutive Photomorphogenic1.

    PubMed

    Meng, Lai-Sheng

    2015-04-01

    ANGUSTIFOLIA3 (AN3), a transcription coactivator, is implicated in modulating cell proliferation. In this study, I found that AN3 is a novel regulator of anthocyanin biosynthesis and light-induced root elongation. Seedlings and seeds lacking AN3 activity presented significantly reduced anthocyanin accumulation and light-induced root elongation, whereas those of transgenic plants harbouring the 35S:AN3 construct exhibited increased anthocyanin accumulation. AN3 is required for the proper expression of other genes that affect anthocyanin accumulation and light-induced root elongation, Constitutive Photomorphogenic1 (COP1), encoding a RING motif - containing E3 ubiquitin ligase. AN3 was associated with COP1 promoter in vivo. Thus, AN3 may act with other proteins that bind to COP1 promoter to promote anthocyanin accumulation and inhibit light-induced root elongation. © 2014 John Wiley & Sons Ltd.

  20. Arsenic Detoxification by Geobacter Species.

    PubMed

    Dang, Yan; Walker, David J F; Vautour, Kaitlin E; Dixon, Steven; Holmes, Dawn E

    2017-02-15

    Insight into the mechanisms for arsenic detoxification by Geobacter species is expected to improve the understanding of global cycling of arsenic in iron-rich subsurface sedimentary environments. Analysis of 14 different Geobacter genomes showed that all of these species have genes coding for an arsenic detoxification system (ars operon), and several have genes required for arsenic respiration (arr operon) and methylation (arsM). Genes encoding four arsenic repressor-like proteins were detected in the genome of G. sulfurreducens; however, only one (ArsR1) regulated transcription of the ars operon. Elimination of arsR1 from the G. sulfurreducens chromosome resulted in enhanced transcription of genes coding for the arsenic efflux pump (Acr3) and arsenate reductase (ArsC). When the gene coding for Acr3 was deleted, cells were not able to grow in the presence of either the oxidized or reduced form of arsenic, while arsC deletion mutants could grow in the presence of arsenite but not arsenate. These studies shed light on how Geobacter influences arsenic mobility in anoxic sediments and may help us develop methods to remediate arsenic contamination in the subsurface. This study examines arsenic transformation mechanisms utilized by Geobacter, a genus of iron-reducing bacteria that are predominant in many anoxic iron-rich subsurface environments. Geobacter species play a major role in microbially mediated arsenic release from metal hydroxides in the subsurface. This release raises arsenic concentrations in drinking water to levels that are high enough to cause major health problems. Therefore, information obtained from studies of Geobacter should shed light on arsenic cycling in iron-rich subsurface sedimentary environments, which may help reduce arsenic-associated illnesses. These studies should also help in the development of biosensors that can be used to detect arsenic contaminants in anoxic subsurface environments. We examined 14 different Geobacter genomes and found that all of these species possess genes coding for an arsenic detoxification system (ars operon), and some also have genes required for arsenic respiration (arr operon) and arsenic methylation (arsM). Copyright © 2017 American Society for Microbiology.

  1. Arsenic Detoxification by Geobacter Species

    PubMed Central

    Walker, David J. F.; Vautour, Kaitlin E.; Dixon, Steven

    2016-01-01

    ABSTRACT Insight into the mechanisms for arsenic detoxification by Geobacter species is expected to improve the understanding of global cycling of arsenic in iron-rich subsurface sedimentary environments. Analysis of 14 different Geobacter genomes showed that all of these species have genes coding for an arsenic detoxification system (ars operon), and several have genes required for arsenic respiration (arr operon) and methylation (arsM). Genes encoding four arsenic repressor-like proteins were detected in the genome of G. sulfurreducens; however, only one (ArsR1) regulated transcription of the ars operon. Elimination of arsR1 from the G. sulfurreducens chromosome resulted in enhanced transcription of genes coding for the arsenic efflux pump (Acr3) and arsenate reductase (ArsC). When the gene coding for Acr3 was deleted, cells were not able to grow in the presence of either the oxidized or reduced form of arsenic, while arsC deletion mutants could grow in the presence of arsenite but not arsenate. These studies shed light on how Geobacter influences arsenic mobility in anoxic sediments and may help us develop methods to remediate arsenic contamination in the subsurface. IMPORTANCE This study examines arsenic transformation mechanisms utilized by Geobacter, a genus of iron-reducing bacteria that are predominant in many anoxic iron-rich subsurface environments. Geobacter species play a major role in microbially mediated arsenic release from metal hydroxides in the subsurface. This release raises arsenic concentrations in drinking water to levels that are high enough to cause major health problems. Therefore, information obtained from studies of Geobacter should shed light on arsenic cycling in iron-rich subsurface sedimentary environments, which may help reduce arsenic-associated illnesses. These studies should also help in the development of biosensors that can be used to detect arsenic contaminants in anoxic subsurface environments. We examined 14 different Geobacter genomes and found that all of these species possess genes coding for an arsenic detoxification system (ars operon), and some also have genes required for arsenic respiration (arr operon) and arsenic methylation (arsM). PMID:27940542

  2. Virtual communities of practice: can they support the prevention agenda in public health?

    PubMed Central

    Ford, Jennifer; Korjonen, Helena; Keswani, Asha; Hughes, Emma

    2015-01-01

    Background Virtual Communities of Practice (CoPs) are flexible communication and knowledge management tools enabling collaboration, sharing of best practice and professional development. There have been few studies that have looked at the use and usefulness of virtual CoPs in public health. Methods This project sought to gather the evidence and develop recommendations for the value of virtual CoPs in public health through a literature review, and through piloting two CoPs in obesity. The research aimed to find out how useful CoPs are in obesity prevention, what makes a CoP successful and what evaluation methods are appropriate. Results CoPs are composed of observers, passive and active contributors with a small group of 'super-users'. All users learn through reading and listening, even if they do not post. The CoPs had higher levels of reading activity as opposed to low levels of posting activity. Longer existence of CoPs usually means more active membership. There are complex reasons why users fail to engage in knowledge sharing. Success of a CoP is creating an online environment where users feel comfortable. CoPs need administrative support and facilitation. Champions play a vital role. Conclusions Evidence shows some encouraging results about the value of CoPs in enabling collaboration and information sharing. Despite low membership numbers of the obesity CoPs piloted, members see value and suggest improvements. Findings suggest that success comes from leadership, champions, and larger networks with more posting activity. Mixed methods of quantitative and qualitative research are appropriate in measuring the use and impact of CoPs. PMID:26284150

  3. 30 CFR 285.630 - [Reserved

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... OF EXISTING FACILITIES ON THE OUTER CONTINENTAL SHELF Plans and Information Requirements Contents of the Construction and Operations Plan § 285.630 [Reserved] Activities Under an Approved COP ...

  4. Hypermutation in derepressed operons of Escherichia coli K12

    PubMed Central

    Wright, Barbara E.; Longacre, Angelika; Reimers, Jacqueline M.

    1999-01-01

    This article presents evidence that starvation for leucine in an Escherichia coli auxotroph triggers metabolic activities that specifically target the leu operon for derepression, increased rates of transcription, and mutation. Derepression of the leu operon was a prerequisite for its activation by the signal nucleotide, guanosine tetraphosphate, which accumulates in response to nutritional stress (the stringent response). A quantitative correlation was established between leuB mRNA abundance and leuB− reversion rates. To further demonstrate that derepression increased mutation rates, the chromosomal leu operon was placed under the control of the inducible tac promoter. When the leu operon was induced by isopropyl-d-thiogalactoside, both leuB mRNA abundance and leuB− reversion rates increased. These investigations suggest that guanosine tetraphosphate may contribute as much as attenuation in regulating leu operon expression and that higher rates of mutation are specifically associated with the derepressed leu operon. PMID:10220423

  5. Conversion from CUL4-based COP1–SPA E3 apparatus to UVR8–COP1–SPA complexes underlies a distinct biochemical function of COP1 under UV-B

    PubMed Central

    Huang, Xi; Ouyang, Xinhao; Yang, Panyu; Lau, On Sun; Chen, Liangbi; Wei, Ning; Deng, Xing Wang

    2013-01-01

    The evolutionarily conserved CONSTITUTIVE PHOTOMORPHOGENESIS 1 (COP1) is a RING and WD40 protein that functions as a substrate receptor of CULLIN4–DAMAGED DNA BINDING PROTEIN 1 (CUL4–DDB1)–based E3 ubiquitin ligases in both plants and animals. In Arabidopsis, COP1 is a central repressor of photomorphogenesis in the form of COP1–SUPPRESSOR OF PHYA (SPA) complex(es). CUL4–DDB1–COP1–SPA suppresses the photomorphogenic program by targeting the transcription factor ELONGATED HYPOCOTYL 5 for degradation. Intriguingly, under photomorphogenic UV-B light, COP1 reverses its repressive role and promotes photomorphogenesis. However, the mechanism by which COP1 is functionally switched is still obscure. Here, we demonstrate that UV-B triggers the physical and functional disassociation of the COP1–SPA core complex(es) from CUL4–DDB1 and the formation of a unique complex(es) containing the UV-B receptor UV RESISTANCE LOCUS 8 (UVR8). The establishment of this UV-B–dependent COP1 complex(es) is associated with its positive modulation of ELONGATED HYPOCOTYL 5 stability and activity, which sheds light on the mechanism of COP1’s promotive action in UV-B–induced photomorphogenesis. PMID:24067658

  6. Cross-Regulation between the phz1 and phz2 Operons Maintain a Balanced Level of Phenazine Biosynthesis in Pseudomonas aeruginosa PAO1

    PubMed Central

    Jiang, Bei; Xiao, Bo; Liu, Linde; Ge, Yihe; Hu, Xiaomei

    2016-01-01

    Gene duplication often provides selective advantages for the survival of microorganisms in adapting to varying environmental conditions. P. aeruginosa PAO1 possesses two seven-gene operons [phz1 (phzA1B1C1D1E1F1G1) and phz2 (phzA2B2C2D2E2F2G2)] that are involved in the biosynthesis of phenazine-1-carboxylic acid and its derivatives. Although the two operons are highly homologous and their functions are well known, it is unclear how the two phz operons coordinate their expressions to maintain the phenazine biosynthesis. By constructing single and double deletion mutants of the two phz operons, we found that the phz1-deletion mutant produced the same or less amount of phenazine-1-carboxylic acid and pyocyanin in GA medium than the phz2-knockout mutant while the phz1-phz2 double knockout mutant did not produce any phenazines. By generating phzA1 and phzA2 translational and transcriptional fusions with a truncated lacZ reporter, we found that the expression of the phz1 operon increased significantly at the post-transcriptional level and did not alter at the transcriptional level in the absence of the phz2 operon. Surprisingly, the expression the phz2 operon increased significantly at the post-transcriptional level and only moderately at the transcriptional level in the absence of the phz1 operon. Our findings suggested that a complex cross-regulation existed between the phz1 and phz2 operons. By mediating the upregulation of one phz operon expression while the other was deleted, this crosstalk would maintain the homeostatic balance of phenazine biosynthesis in P. aeruginosa PAO1. PMID:26735915

  7. Genome-wide analysis of trans-splicing in the nematode Pristionchus pacificus unravels conserved gene functions for germline and dauer development in divergent operons.

    PubMed

    Sinha, Amit; Langnick, Claudia; Sommer, Ralf J; Dieterich, Christoph

    2014-09-01

    Discovery of trans-splicing in multiple metazoan lineages led to the identification of operon-like gene organization in diverse organisms, including trypanosomes, tunicates, and nematodes, but the functional significance of such operons is not completely understood. To see whether the content or organization of operons serves similar roles across species, we experimentally defined operons in the nematode model Pristionchus pacificus. We performed affinity capture experiments on mRNA pools to specifically enrich for transcripts that are trans-spliced to either the SL1- or SL2-spliced leader, using spliced leader-specific probes. We obtained distinct trans-splicing patterns from the analysis of three mRNA pools (total mRNA, SL1 and SL2 fraction) by RNA-seq. This information was combined with a genome-wide analysis of gene orientation and spacing. We could confirm 2219 operons by RNA-seq data out of 6709 candidate operons, which were predicted by sequence information alone. Our gene order comparison of the Caenorhabditis elegans and P. pacificus genomes shows major changes in operon organization in the two species. Notably, only 128 out of 1288 operons in C. elegans are conserved in P. pacificus. However, analysis of gene-expression profiles identified conserved functions such as an enrichment of germline-expressed genes and higher expression levels of operonic genes during recovery from dauer arrest in both species. These results provide support for the model that a necessity for increased transcriptional efficiency in the context of certain developmental processes could be a selective constraint for operon evolution in metazoans. Our method is generally applicable to other metazoans to see if similar functional constraints regulate gene organization into operons. © 2014 Sinha et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.

  8. Immunomodulation of Experimental Autoimmune Encephalomyelitis by Oral Administration of Copolymer 1

    NASA Astrophysics Data System (ADS)

    Teitelbaum, Dvora; Arnon, Ruth; Sela, Michael

    1999-03-01

    The activity of copolymer 1 (Cop 1, Copax-one, glatiramer acetate) in suppressing experimental autoimmune encephalomyelitis (EAE) and in the treatment of multiple sclerosis patients when injected parenterally has been extensively demonstrated. In the present study we addressed the question of whether Cop 1 can induce oral tolerance to EAE similar to myelin basic protein (MBP). We now have demonstrated that oral Cop 1 inhibited EAE induction in both rats and mice. Furthermore, oral Cop 1 was more effective than oral MBP in suppressing EAE in rats. The beneficial effect of oral Cop 1 was found to be associated with specific inhibition of the proliferative and Th1 cytokine secretion responses to MBP of spleen cells from Cop 1-fed mice and rats. In all of these assays, oral Cop 1 was more effective than oral MBP. The tolerance induced by Cop 1 could be adoptively transferred with spleen cells from Cop 1-fed animals. Furthermore, Cop 1-specific T cell lines, which inhibit EAE induction in vivo, could be isolated from the above spleen cells. These T cell lines secrete the anti-inflammatory cytokines IL-10 and transforming growth factor type β , but not IL-4, in response to both Cop 1 and MBP. In conclusion, oral Cop 1 has a beneficial effect on the development of EAE that is associated with down-regulation of T cell immune responses to MBP and is mediated by Th2/3 type regulatory cells. These results suggest that oral administration of Cop 1 may modulate multiple sclerosis as well.

  9. Long-term prognosis of patients with carbon monoxide poisoning: a nationwide cohort study.

    PubMed

    Huang, Chien-Cheng; Chung, Min-Hsien; Weng, Shih-Feng; Chien, Chih-Chiang; Lin, Shio-Jean; Lin, Hung-Jung; Guo, How-Ran; Su, Shih-Bin; Hsu, Chien-Chin; Juan, Chi-Wen

    2014-01-01

    Carbon monoxide poisoning (COP) often produces severe complications and can be fatal. Because this topic has not been well delineated, we investigated long-term prognoses of patients with COP (COP[+]). In this retrospective nationwide cohort study, 441 COP[+] patients and 8820 COP[-] controls (120) from 1999 to 2010 were selected from Taiwan's National Health Insurance Research Database. Thirty-seven (8.39%) COP[+] patients and 142 (1.61%) controls died (P<0.0001) during follow-up. Incidence rate ratios (IRR) of death were 5.24 times higher in COP[+] patients than in controls (P<0.0001). The risk of death was particularly high in the first month after COP (IRR: 308.78; 95% confidence interval [CI]: 40.79-2337.56), 1 to 6 months after (IRR: 18.92; 95% CI: 7.69-46.56), and 6-12 months after (IRR: 4.73; 95% CI: 1.02-21.90). After adjusting for age, gender, and selected comorbidities, the hazard ratio of death for COP[+] patients was still 4.097 times higher than for controls. Moreover, older age (≥30 years old), male gender, diabetes mellitus, hypertension, and low income were also independent mortality predictors. COP significantly increases the risk for long-term mortality. Early follow-up and secondary prevention of death are needed for patients with COP.

  10. Facile chemical synthesis and equilibrium unfolding properties of CopG

    PubMed Central

    Wales, Thomas E.; Richardson, Jane S.; Fitzgerald, Michael C.

    2004-01-01

    The 45-amino acid polypeptide chain of the homodimeric transcriptional repressor, CopG, was chemically synthesized by stepwise solid phase peptide synthesis (SPPS) using a protocol based on Boc-chemistry. The product obtained from the synthesis was readily purified by reversed-phase HPLC to give a good overall yield (21% by weight). Moreover, the synthetic CopG constructs prepared in this work folded into three-dimensional structures similar to the wild-type protein prepared using conventional recombinant methods as judged by far UV-CD spectroscopy. A fluorescent CopG analog, (Y39W)CopG, was also designed and chemically synthesized to facilitate biophysical studies of CopG’s protein folding and assembly reaction. The guanidinium chloride-induced equilibrium unfolding properties of the wild-type CopG and (Y39W)CopG constructs in this work were characterized and used to develop a model for CopG’s equilibrium unfolding reaction. Our results indicate that CopG’s folding and assembly reaction is well modeled by a two-state process involving folded dimer and unfolded monomer. Using this model, ΔGf and m-values of −13.42 ± 0.04 kcal/mole dimer and 1.92 ± 0.01 kcal/(mole M) were calculated for CopG. PMID:15169951

  11. The pap Operon of Avian Pathogenic Escherichia coli Strain O1:K1 Is Located on a Novel Pathogenicity Island

    PubMed Central

    Kariyawasam, Subhashinie; Johnson, Timothy J.; Nolan, Lisa K.

    2006-01-01

    We have identified a 56-kb pathogenicity island (PAI) in avian pathogenic Escherichia coli strain O1:K1 (APEC-O1). This PAI, termed PAI IAPEC-O1, is integrated adjacent to the 3′ end of the pheV tRNA gene. It carries putative virulence genes of APEC (pap operon), other E. coli genes (tia and ireA), and a 1.5-kb region unique to APEC-O1. The kps gene cluster required for the biosynthesis of polysialic acid capsule was mapped to a location immediately downstream of this PAI. PMID:16369033

  12. 30 CFR 585.628 - How will BOEM process my COP?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 30 Mineral Resources 2 2014-07-01 2014-07-01 false How will BOEM process my COP? 585.628 Section... COP? (a) BOEM will review your submitted COP, and the information provided pursuant to § 585.627, to... reviews. We will notify you if your submitted COP lacks any necessary information. (b) BOEM will prepare...

  13. 30 CFR 585.628 - How will BOEM process my COP?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 30 Mineral Resources 2 2013-07-01 2013-07-01 false How will BOEM process my COP? 585.628 Section... COP? (a) BOEM will review your submitted COP, and the information provided pursuant to § 585.627, to... reviews. We will notify you if your submitted COP lacks any necessary information. (b) BOEM will prepare...

  14. 30 CFR 585.628 - How will BOEM process my COP?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 30 Mineral Resources 2 2012-07-01 2012-07-01 false How will BOEM process my COP? 585.628 Section... COP? (a) BOEM will review your submitted COP, and the information provided pursuant to § 585.627, to... reviews. We will notify you if your submitted COP lacks any necessary information. (b) BOEM will prepare...

  15. Problem-Solving Test: Tryptophan Operon Mutants

    ERIC Educational Resources Information Center

    Szeberenyi, Jozsef

    2010-01-01

    This paper presents a problem-solving test that deals with the regulation of the "trp" operon of "Escherichia coli." Two mutants of this operon are described: in mutant A, the operator region of the operon carries a point mutation so that it is unable to carry out its function; mutant B expresses a "trp" repressor protein unable to bind…

  16. Gene context conservation of a higher order than operons.

    PubMed

    Lathe, W C; Snel, B; Bork, P

    2000-10-01

    Operons, co-transcribed and co-regulated contiguous sets of genes, are poorly conserved over short periods of evolutionary time. The gene order, gene content and regulatory mechanisms of operons can be very different, even in closely related species. Here, we present several lines of evidence which suggest that, although an operon and its individual genes and regulatory structures are rearranged when comparing the genomes of different species, this rearrangement is a conservative process. Genomic rearrangements invariably maintain individual genes in very specific functional and regulatory contexts. We call this conserved context an uber-operon.

  17. Regulation of the yjjQ-bglJ Operon, Encoding LuxR-Type Transcription Factors, and the Divergent yjjP Gene by H-NS and LeuO▿ †

    PubMed Central

    Stratmann, Thomas; Madhusudan, S.; Schnetz, Karin

    2008-01-01

    The yjjQ and bglJ genes encode LuxR-type transcription factors conserved in several enterobacterial species. YjjQ is a potential virulence factor in avian pathogenic Escherichia coli. BglJ counteracts the silencing of the bgl (β-glucoside) operon by H-NS in E. coli K-12. Here we show that yjjQ and bglJ form an operon carried by E. coli K-12, whose expression is repressed by the histone-like nucleoid structuring (H-NS) protein. The LysR-type transcription factor LeuO counteracts this repression. Furthermore, the yjjP gene, encoding a membrane protein of unknown function and located upstream in divergent orientation to the yjjQ-bglJ operon, is likewise repressed by H-NS. Mapping of the promoters as well as the H-NS and LeuO binding sites within the 555-bp intergenic region revealed that H-NS binds to the center of the AT-rich regulatory region and distal to the divergent promoters. LeuO sites map to the center and to positions distal to the yjjQ promoters, while one LeuO binding site overlaps with the divergent yjjP promoter. This latter LeuO site is required for full derepression of the yjjQ promoters. The arrangement of regulatory sites suggests that LeuO restructures the nucleoprotein complex formed by H-NS. Furthermore, the data support the conclusion that LeuO, whose expression is likewise repressed by H-NS and which is a virulence factor in Salmonella enterica, is a master regulator that among other loci, also controls the yjjQ-bglJ operon and thus indirectly the presumptive targets of YjjQ and BglJ. PMID:18055596

  18. Development and validation of an rDNA operon based primer walking strategy applicable to de novo bacterial genome finishing

    PubMed Central

    Eastman, Alexander W.; Yuan, Ze-Chun

    2015-01-01

    Advances in sequencing technology have drastically increased the depth and feasibility of bacterial genome sequencing. However, little information is available that details the specific techniques and procedures employed during genome sequencing despite the large numbers of published genomes. Shotgun approaches employed by second-generation sequencing platforms has necessitated the development of robust bioinformatics tools for in silico assembly, and complete assembly is limited by the presence of repetitive DNA sequences and multi-copy operons. Typically, re-sequencing with multiple platforms and laborious, targeted Sanger sequencing are employed to finish a draft bacterial genome. Here we describe a novel strategy based on the identification and targeted sequencing of repetitive rDNA operons to expedite bacterial genome assembly and finishing. Our strategy was validated by finishing the genome of Paenibacillus polymyxa strain CR1, a bacterium with potential in sustainable agriculture and bio-based processes. An analysis of the 38 contigs contained in the P. polymyxa strain CR1 draft genome revealed 12 repetitive rDNA operons with varied intragenic and flanking regions of variable length, unanimously located at contig boundaries and within contig gaps. These highly similar but not identical rDNA operons were experimentally verified and sequenced simultaneously with multiple, specially designed primer sets. This approach also identified and corrected significant sequence rearrangement generated during the initial in silico assembly of sequencing reads. Our approach reduces the required effort associated with blind primer walking for contig assembly, increasing both the speed and feasibility of genome finishing. Our study further reinforces the notion that repetitive DNA elements are major limiting factors for genome finishing. Moreover, we provided a step-by-step workflow for genome finishing, which may guide future bacterial genome finishing projects. PMID:25653642

  19. 30 CFR 285.634 - What activities require a revision to my COP, and when will MMS approve the revision?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... ENERGY MANAGEMENT, REGULATION, AND ENFORCEMENT, DEPARTMENT OF THE INTERIOR OFFSHORE RENEWABLE ENERGY... location of your onshore support base from one State to another or to a new base requiring expansion; (6...

  20. On-board attitude determination for the Explorer Platform satellite

    NASA Technical Reports Server (NTRS)

    Jayaraman, C.; Class, B.

    1992-01-01

    This paper describes the attitude determination algorithm for the Explorer Platform satellite. The algorithm, which is baselined on the Landsat code, is a six-element linear quadratic state estimation processor, in the form of a Kalman filter augmented by an adaptive filter process. Improvements to the original Landsat algorithm were required to meet mission pointing requirements. These consisted of a more efficient sensor processing algorithm and the addition of an adaptive filter which acts as a check on the Kalman filter during satellite slew maneuvers. A 1750A processor will be flown on board the satellite for the first time as a coprocessor (COP) in addition to the NASA Standard Spacecraft Computer. The attitude determination algorithm, which will be resident in the COP's memory, will make full use of its improved processing capabilities to meet mission requirements. Additional benefits were gained by writing the attitude determination code in Ada.

  1. 20 CFR 10.221 - How is a claim for COP controverted?

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 20 Employees' Benefits 1 2014-04-01 2012-04-01 true How is a claim for COP controverted? 10.221... AMENDED Continuation of Pay Controversion and Termination of Cop § 10.221 How is a claim for COP... must controvert the claim for COP on Form CA-1, explaining in detail the basis for the refusal. The...

  2. 20 CFR 10.221 - How is a claim for COP controverted?

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 20 Employees' Benefits 1 2010-04-01 2010-04-01 false How is a claim for COP controverted? 10.221... AMENDED Continuation of Pay Controversion and Termination of Cop § 10.221 How is a claim for COP... must controvert the claim for COP on Form CA-1, explaining in detail the basis for the refusal. The...

  3. 20 CFR 10.216 - How is the pay rate for COP calculated?

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 20 Employees' Benefits 1 2012-04-01 2012-04-01 false How is the pay rate for COP calculated? 10..., AS AMENDED Continuation of Pay Calculation of Cop § 10.216 How is the pay rate for COP calculated? The employer shall calculate COP using the period of time and the weekly pay rate. (a) The pay rate...

  4. 20 CFR 10.221 - How is a claim for COP controverted?

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 20 Employees' Benefits 1 2011-04-01 2011-04-01 false How is a claim for COP controverted? 10.221... AMENDED Continuation of Pay Controversion and Termination of Cop § 10.221 How is a claim for COP... must controvert the claim for COP on Form CA-1, explaining in detail the basis for the refusal. The...

  5. 20 CFR 10.221 - How is a claim for COP controverted?

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 20 Employees' Benefits 1 2013-04-01 2012-04-01 true How is a claim for COP controverted? 10.221... AMENDED Continuation of Pay Controversion and Termination of Cop § 10.221 How is a claim for COP... must controvert the claim for COP on Form CA-1, explaining in detail the basis for the refusal. The...

  6. 20 CFR 10.215 - How does OWCP compute the number of days of COP used?

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... COP used? 10.215 Section 10.215 Employees' Benefits OFFICE OF WORKERS' COMPENSATION PROGRAMS...' COMPENSATION ACT, AS AMENDED Continuation of Pay Calculation of Cop § 10.215 How does OWCP compute the number of days of COP used? COP is payable for a maximum of 45 calendar days, and every day used is counted...

  7. 20 CFR 10.221 - How is a claim for COP controverted?

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 20 Employees' Benefits 1 2012-04-01 2012-04-01 false How is a claim for COP controverted? 10.221... AMENDED Continuation of Pay Controversion and Termination of Cop § 10.221 How is a claim for COP... must controvert the claim for COP on Form CA-1, explaining in detail the basis for the refusal. The...

  8. 20 CFR 10.215 - How does OWCP compute the number of days of COP used?

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... COP used? 10.215 Section 10.215 Employees' Benefits OFFICE OF WORKERS' COMPENSATION PROGRAMS...' COMPENSATION ACT, AS AMENDED Continuation of Pay Calculation of Cop § 10.215 How does OWCP compute the number of days of COP used? COP is payable for a maximum of 45 calendar days, and every day used is counted...

  9. 20 CFR 10.215 - How does OWCP compute the number of days of COP used?

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... COP used? 10.215 Section 10.215 Employees' Benefits OFFICE OF WORKERS' COMPENSATION PROGRAMS...' COMPENSATION ACT, AS AMENDED Continuation of Pay Calculation of Cop § 10.215 How does OWCP compute the number of days of COP used? COP is payable for a maximum of 45 calendar days, and every day used is counted...

  10. 20 CFR 10.215 - How does OWCP compute the number of days of COP used?

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... COP used? 10.215 Section 10.215 Employees' Benefits OFFICE OF WORKERS' COMPENSATION PROGRAMS...' COMPENSATION ACT, AS AMENDED Continuation of Pay Calculation of Cop § 10.215 How does OWCP compute the number of days of COP used? COP is payable for a maximum of 45 calendar days, and every day used is counted...

  11. Role of the parCBA Operon of the Broad-Host-Range Plasmid RK2 in Stable Plasmid Maintenance

    PubMed Central

    Easter, Carla L.; Schwab, Helmut; Helinski, Donald R.

    1998-01-01

    The par region of the stably maintained broad-host-range plasmid RK2 is organized as two divergent operons, parCBA and parDE, and a cis-acting site. parDE encodes a postsegregational killing system, and parCBA encodes a resolvase (ParA), a nuclease (ParB), and a protein of unknown function (ParC). The present study was undertaken to further delineate the role of the parCBA region in the stable maintenance of RK2 by first introducing precise deletions in the three genes and then assessing the abilities of the different constructs to stabilize RK2 in three strains of Escherichia coli and two strains of Pseudomonas aeruginosa. The intact parCBA operon was effective in stabilizing a conjugation-defective RK2 derivative in E. coli MC1061K and RR1 but was relatively ineffective in E. coli MV10Δlac. In the two strains in which the parCBA operon was effective, deletions in parB, parC, or both parB and parC caused an approximately twofold reduction in the stabilizing ability of the operon, while a deletion in the parA gene resulted in a much greater loss of parCBA activity. For P. aeruginosa PAO1161Rifr, the parCBA operon provided little if any plasmid stability, but for P. aeruginosa PAC452Rifr, the RK2 plasmid was stabilized to a substantial extent by parCBA. With this latter strain, parA and res alone were sufficient for stabilization. The cer resolvase system of plasmid ColE1 and the loxP/Cre system of plasmid P1 were tested in comparison with the parCBA operon. We found that, not unlike what was previously observed with MC1061K, cer failed to stabilize the RK2 plasmid with par deletions in strain MV10Δlac, but this multimer resolution system was effective in stabilizing the plasmid in strain RR1. The loxP/Cre system, on the other hand, was very effective in stabilizing the plasmid in all three E. coli strains. These observations indicate that the parA gene, along with its res site, exhibits a significant level of plasmid stabilization in the absence of the parC and parB genes but that in at least one E. coli strain, all three genes are required for maximum stabilization. It cannot be determined from these results whether or not the stabilization effects seen with parCBA or the cer and loxP/Cre systems are strictly due to a reduction in the level of RK2 dimers and an increase in the number of plasmid monomer units or if these systems play a role in a more complex process of plasmid stabilization that requires as an essential step the resolution of plasmid dimers. PMID:9811663

  12. Cultivating a Network For Messaging About Climate Change Across an Urban System

    NASA Astrophysics Data System (ADS)

    Wertheim, J.

    2014-12-01

    Currently, some of the most promising efforts to address climate change are taking place at the scale of cities and municipalities. Large urban areas host an active population of organizations working to influence local environmental policies more rigorous than those at the state and national level. The composition of these groups is broadening as impacts of climate change are being recognized as relevant to more sectors within urban systems, from health centers to community leaders, leading more organizations to consider how they can raise awareness and gain support for their needs. The National Geographic Society, as part of the National Science Foundation (NSF)-funded Climate and Urban Systems Partnership (CUSP), has convened a pilot "community of practice" (CoP) consisting of organizations working at the local level in Washington, DC to communicate with audiences, from the general public to local government agencies, about ways that climate change is predicted to affect the city and what can be done about it. The purpose of the CoP was initially to help these groups coordinate their activities, share knowledge and resources, and to create a platform for ongoing collaborative learning. While the CoP is still evolving, it is clear that it has potential to provide even deeper and more meaningful support to these groups' efforts. Developing effective messaging about climate change across an urban system depends on the valuable insight these groups have into their audience's interests, beliefs, and knowledge, but it also requires a set of competencies that few members of the CoP hold. As conveners of the CoP, we have identified and prioritized those competencies and are developing a process for training CoP members to apply their expertise to implement empirically-based best practices in climate change messaging, public communication, and integration of data and visualizations. The process of training the group has the potential to both create a CoP that becomes a trusted resource for climate messaging activities in DC and an enduring network of professionals who identify participating in the group's ongoing work as essential to their own professional development

  13. Molecular Characterization of Copper Resistance Genes from Xanthomonas citri subsp. citri and Xanthomonas alfalfae subsp. citrumelonis▿

    PubMed Central

    Behlau, Franklin; Canteros, Blanca I.; Minsavage, Gerald V.; Jones, Jeffrey B.; Graham, James H.

    2011-01-01

    Copper sprays have been widely used for control of endemic citrus canker caused by Xanthomonas citri subsp. citri in citrus-growing areas for more than 2 decades. Xanthomonas alfalfae subsp. citrumelonis populations were also exposed to frequent sprays of copper for several years as a protective measure against citrus bacterial spot (CBS) in Florida citrus nurseries. Long-term use of these bactericides has led to the development of copper-resistant (Cur) strains in both X. citri subsp. citri and X. alfalfae subsp. citrumelonis, resulting in a reduction of disease control. The objectives of this study were to characterize for the first time the genetics of copper resistance in X. citri subsp. citri and X. alfalfae subsp. citrumelonis and to compare these organisms to other Cur bacteria. Copper resistance determinants from X. citri subsp. citri strain A44(pXccCu2) from Argentina and X. alfalfae subsp. citrumelonis strain 1381(pXacCu2) from Florida were cloned and sequenced. Open reading frames (ORFs) related to the genes copL, copA, copB, copM, copG, copC, copD, and copF were identified in X. citri subsp. citri A44. The same ORFs, except copC and copD, were also present in X. alfalfae subsp. citrumelonis 1381. Transposon mutagenesis of the cloned copper resistance determinants in pXccCu2 revealed that copper resistance in X. citri subsp. citri strain A44 is mostly due to copL, copA, and copB, which are the genes in the cloned cluster with the highest nucleotide homology (≥92%) among different Cur bacteria. PMID:21515725

  14. A phylogenomic analysis of the Actinomycetales mce operons.

    PubMed

    Casali, Nicola; Riley, Lee W

    2007-02-26

    The genome of Mycobacterium tuberculosis harbors four copies of a cluster of genes termed mce operons. Despite extensive research that has demonstrated the importance of these operons on infection outcome, their physiological function remains obscure. Expanding databases of complete microbial genome sequences facilitate a comparative genomic approach that can provide valuable insight into the role of uncharacterized proteins. The M. tuberculosis mce loci each include two yrbE and six mce genes, which have homology to ABC transporter permeases and substrate-binding proteins, respectively. Operons with an identical structure were identified in all Mycobacterium species examined, as well as in five other Actinomycetales genera. Some of the Actinomycetales mce operons include an mkl gene, which encodes an ATPase resembling those of ABC uptake transporters. The phylogenetic profile of Mkl orthologs exactly matched that of the Mce and YrbE proteins. Through topology and motif analyses of YrbE homologs, we identified a region within the penultimate cytoplasmic loop that may serve as the site of interaction with the putative cognate Mkl ATPase. Homologs of the exported proteins encoded adjacent to the M. tuberculosis mce operons were detected in a conserved chromosomal location downstream of the majority of Actinomycetales operons. Operons containing linked mkl, yrbE and mce genes, resembling the classic organization of an ABC importer, were found to be common in Gram-negative bacteria and appear to be associated with changes in properties of the cell surface. Evidence presented suggests that the mce operons of Actinomycetales species and related operons in Gram-negative bacteria encode a subfamily of ABC uptake transporters with a possible role in remodeling the cell envelope.

  15. High accuracy operon prediction method based on STRING database scores.

    PubMed

    Taboada, Blanca; Verde, Cristina; Merino, Enrique

    2010-07-01

    We present a simple and highly accurate computational method for operon prediction, based on intergenic distances and functional relationships between the protein products of contiguous genes, as defined by STRING database (Jensen,L.J., Kuhn,M., Stark,M., Chaffron,S., Creevey,C., Muller,J., Doerks,T., Julien,P., Roth,A., Simonovic,M. et al. (2009) STRING 8-a global view on proteins and their functional interactions in 630 organisms. Nucleic Acids Res., 37, D412-D416). These two parameters were used to train a neural network on a subset of experimentally characterized Escherichia coli and Bacillus subtilis operons. Our predictive model was successfully tested on the set of experimentally defined operons in E. coli and B. subtilis, with accuracies of 94.6 and 93.3%, respectively. As far as we know, these are the highest accuracies ever obtained for predicting bacterial operons. Furthermore, in order to evaluate the predictable accuracy of our model when using an organism's data set for the training procedure, and a different organism's data set for testing, we repeated the E. coli operon prediction analysis using a neural network trained with B. subtilis data, and a B. subtilis analysis using a neural network trained with E. coli data. Even for these cases, the accuracies reached with our method were outstandingly high, 91.5 and 93%, respectively. These results show the potential use of our method for accurately predicting the operons of any other organism. Our operon predictions for fully-sequenced genomes are available at http://operons.ibt.unam.mx/OperonPredictor/.

  16. The Anaerobe-Specific Orange Protein Complex of Desulfovibrio vulgaris Hildenborough Is Encoded by Two Divergent Operons Coregulated by σ54 and a Cognate Transcriptional Regulator▿†

    PubMed Central

    Fiévet, Anouchka; My, Laetitia; Cascales, Eric; Ansaldi, Mireille; Pauleta, Sofia R.; Moura, Isabel; Dermoun, Zorah; Bernard, Christophe S.; Dolla, Alain; Aubert, Corinne

    2011-01-01

    Analysis of sequenced bacterial genomes revealed that the genomes encode more than 30% hypothetical and conserved hypothetical proteins of unknown function. Among proteins of unknown function that are conserved in anaerobes, some might be determinants of the anaerobic way of life. This study focuses on two divergent clusters specifically found in anaerobic microorganisms and mainly composed of genes encoding conserved hypothetical proteins. We show that the two gene clusters DVU2103-DVU2104-DVU2105 (orp2) and DVU2107-DVU2108-DVU2109 (orp1) form two divergent operons transcribed by the σ54-RNA polymerase. We further demonstrate that the σ54-dependent transcriptional regulator DVU2106, located between orp1 and orp2, collaborates with σ54-RNA polymerase to orchestrate the simultaneous expression of the divergent orp operons. DVU2106, whose structural gene is transcribed by the σ70-RNA polymerase, negatively retrocontrols its own expression. By using an endogenous pulldown strategy, we identify a physiological complex composed of DVU2103, DVU2104, DVU2105, DVU2108, and DVU2109. Interestingly, inactivation of DVU2106, which is required for orp operon transcription, induces morphological defects that are likely linked to the absence of the ORP complex. A putative role of the ORP proteins in positioning the septum during cell division is discussed. PMID:21531797

  17. Using Synthetic Biology to Distinguish and Overcome Regulatory and Functional Barriers Related to Nitrogen Fixation

    PubMed Central

    Wang, Xia; Yang, Jian-Guo; Chen, Li; Wang, Ji-Long; Cheng, Qi; Dixon, Ray; Wang, Yi-Ping

    2013-01-01

    Biological nitrogen fixation is a complex process requiring multiple genes working in concert. To date, the Klebsiella pneumoniae nif gene cluster, divided into seven operons, is one of the most studied systems. Its nitrogen fixation capacity is subject to complex cascade regulation and physiological limitations. In this report, the entire K. pneumoniae nif gene cluster was reassembled as operon-based BioBrick parts in Escherichia coli. It provided ∼100% activity of native K. pneumoniae system. Based on the expression levels of these BioBrick parts, a T7 RNA polymerase–LacI expression system was used to replace the σ54-dependent promoters located upstream of nif operons. Expression patterns of nif operons were critical for the maximum activity of the recombinant system. By mimicking these expression levels with variable-strength T7-dependent promoters, ∼42% of the nitrogenase activity of the σ54-dependent nif system was achieved in E. coli. When the newly constructed T7-dependent nif system was challenged with different genetic and physiological conditions, it bypassed the original complex regulatory circuits, with minor physiological limitations. Therefore, we have successfully replaced the nif regulatory elements with a simple expression system that may provide the first step for further research of introducing nif genes into eukaryotic organelles, which has considerable potentials in agro-biotechnology. PMID:23935879

  18. Solving a discrete model of the lac operon using Z3

    NASA Astrophysics Data System (ADS)

    Gutierrez, Natalia A.

    2014-05-01

    A discrete model for the Lcac Operon is solved using the SMT-solver Z3. Traditionally the Lac Operon is formulated in a continuous math model. This model is a system of ordinary differential equations. Here, it was considerated as a discrete model, based on a Boolean red. The biological problem of Lac Operon is enunciated as a problem of Boolean satisfiability, and it is solved using an STM-solver named Z3. Z3 is a powerful solver that allows understanding the basic dynamic of the Lac Operon in an easier and more efficient way. The multi-stability of the Lac Operon can be easily computed with Z3. The code that solves the Boolean red can be written in Python language or SMT-Lib language. Both languages were used in local version of the program as online version of Z3. For future investigations it is proposed to solve the Boolean red of Lac Operon using others SMT-solvers as cvc4, alt-ergo, mathsat and yices.

  19. Statistics Graduate Students' Professional Development for Teaching: A Communities of Practice Model

    NASA Astrophysics Data System (ADS)

    Justice, Nicola

    Graduate teaching assistants (GTAs) are responsible for instructing approximately 25% of introductory statistics courses in the United States (Blair, Kirkman, & Maxwell, 2013). Most research on GTA professional development focuses on structured activities (e.g., courses, workshops) that have been developed to improve GTAs' pedagogy and content knowledge. Few studies take into account the social contexts of GTAs' professional development. However, GTAs perceive their social interactions with other GTAs to be a vital part of their preparation and support for teaching (e.g., Staton & Darling, 1989). Communities of practice (CoPs) are one way to bring together the study of the social contexts and structured activities of GTA professional development. CoPs are defined as groups of practitioners who deepen their knowledge and expertise by interacting with each other on an ongoing basis (e.g., Lave & Wenger, 1991). Graduate students may participate in CoPs related to teaching in many ways, including attending courses or workshops, participating in weekly meetings, engaging in informal discussions about teaching, or participating in e-mail conversations related to teaching tasks. This study explored the relationship between statistics graduate students' experiences in CoPs and the extent to which they hold student-centered teaching beliefs. A framework for characterizing GTAs' experiences in CoPs was described and a theoretical model relating these characteristics to GTAs' beliefs was developed. To gather data to test the model, the Graduate Students' Experiences Teaching Statistics (GETS) Inventory was created. Items were written to collect information about GTAs' current teaching beliefs, teaching beliefs before entering their degree programs, characteristics of GTAs' experiences in CoPs, and demographic information. Using an online program, the GETS Inventory was administered to N =218 statistics graduate students representing 37 institutions in 24 different U.S. states. The data gathered from the national survey suggest that statistics graduate students often experience CoPs through required meetings and voluntary discussions about teaching. Participants feel comfortable disagreeing with the people they perceive to be most influential on their teaching beliefs. Most participants perceive a faculty member to have the most influential role in shaping their teaching beliefs. The survey data did not provide evidence to support the proposed theoretical model relating characteristics of experiences in CoPs and beliefs about teaching statistics. Based on cross-validation results, prior beliefs about teaching statistics was the best predictor of current beliefs. Additional models were retained that included student characteristics suggested by previous literature to be associated with student-centered or traditional teaching beliefs (e.g., prior teaching experience, international student status). The results of this study can be used to inform future efforts to help promote student-centered teaching beliefs and teaching practices among statistics GTAs. Modifications to the GETS Inventory are suggested for use in future research designed to gather information about GTAs, their teaching beliefs, and their experiences in CoPs. Suggestions are also made for aspects of CoPs that might be studied further in order to learn how CoPs can promote teaching beliefs and practices that support student learning.

  20. Assessing the genetic diversity of Cu resistance in mine tailings through high-throughput recovery of full-length copA genes

    PubMed Central

    Li, Xiaofang; Zhu, Yong-Guan; Shaban, Babak; Bruxner, Timothy J. C.; Bond, Philip L.; Huang, Longbin

    2015-01-01

    Characterizing the genetic diversity of microbial copper (Cu) resistance at the community level remains challenging, mainly due to the polymorphism of the core functional gene copA. In this study, a local BLASTN method using a copA database built in this study was developed to recover full-length putative copA sequences from an assembled tailings metagenome; these sequences were then screened for potentially functioning CopA using conserved metal-binding motifs, inferred by evolutionary trace analysis of CopA sequences from known Cu resistant microorganisms. In total, 99 putative copA sequences were recovered from the tailings metagenome, out of which 70 were found with high potential to be functioning in Cu resistance. Phylogenetic analysis of selected copA sequences detected in the tailings metagenome showed that topology of the copA phylogeny is largely congruent with that of the 16S-based phylogeny of the tailings microbial community obtained in our previous study, indicating that the development of copA diversity in the tailings might be mainly through vertical descent with few lateral gene transfer events. The method established here can be used to explore copA (and potentially other metal resistance genes) diversity in any metagenome and has the potential to exhaust the full-length gene sequences for downstream analyses. PMID:26286020

  1. Long-Term Prognosis of Patients with Carbon Monoxide Poisoning: A Nationwide Cohort Study

    PubMed Central

    Chung, Min-Hsien; Weng, Shih-Feng; Chien, Chih-Chiang; Lin, Shio-Jean; Lin, Hung-Jung; Guo, How-Ran; Hsu, Chien-Chin; Juan, Chi-Wen

    2014-01-01

    Background Carbon monoxide poisoning (COP) often produces severe complications and can be fatal. Because this topic has not been well delineated, we investigated long-term prognoses of patients with COP (COP[+]). Methods In this retrospective nationwide cohort study, 441 COP[+] patients and 8820 COP[−] controls (120) from 1999 to 2010 were selected from Taiwan’s National Health Insurance Research Database. Results Thirty-seven (8.39%) COP[+] patients and 142 (1.61%) controls died (P<0.0001) during follow-up. Incidence rate ratios (IRR) of death were 5.24 times higher in COP[+] patients than in controls (P<0.0001). The risk of death was particularly high in the first month after COP (IRR: 308.78; 95% confidence interval [CI]: 40.79–2337.56), 1 to 6 months after (IRR: 18.92; 95% CI: 7.69–46.56), and 6–12 months after (IRR: 4.73; 95% CI: 1.02–21.90). After adjusting for age, gender, and selected comorbidities, the hazard ratio of death for COP[+] patients was still 4.097 times higher than for controls. Moreover, older age (≥30 years old), male gender, diabetes mellitus, hypertension, and low income were also independent mortality predictors. Conclusions COP significantly increases the risk for long-term mortality. Early follow-up and secondary prevention of death are needed for patients with COP. PMID:25167083

  2. 30 CFR 585.634 - What activities require a revision to my COP, and when will BOEM approve the revision?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... ENERGY MANAGEMENT, DEPARTMENT OF THE INTERIOR OFFSHORE RENEWABLE ENERGY AND ALTERNATE USES OF EXISTING... support base from one State to another or to a new base requiring expansion; (6) Changes in the location...

  3. 30 CFR 585.634 - What activities require a revision to my COP, and when will BOEM approve the revision?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... ENERGY MANAGEMENT, DEPARTMENT OF THE INTERIOR OFFSHORE RENEWABLE ENERGY AND ALTERNATE USES OF EXISTING... support base from one State to another or to a new base requiring expansion; (6) Changes in the location...

  4. 30 CFR 585.634 - What activities require a revision to my COP, and when will BOEM approve the revision?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... ENERGY MANAGEMENT, DEPARTMENT OF THE INTERIOR OFFSHORE RENEWABLE ENERGY AND ALTERNATE USES OF EXISTING... support base from one State to another or to a new base requiring expansion; (6) Changes in the location...

  5. Center-of-pressure movements during equine-assisted activities.

    PubMed

    Clayton, Hilary M; Kaiser, Leeann J; de Pue, Bonnie; Kaiser, Lana

    2011-01-01

    We compared anteroposterior and mediolateral range of motion and velocity of the center of pressure (COP) on the horse's back between riders without disabilities and riders with cerebral palsy. An electronic pressure mat was used to track COP movements beneath the saddle in 4 riders without disabilities and 4 riders with cerebral palsy. Comparisons between rider groups were made using the Mann-Whitney test (p < .05). The two rider groups differed significantly in anteroposterior range of COP motion, mediolateral range of COP motion, and mediolateral COP velocity. Anteroposterior COP velocity did not differ between groups. The results suggest that measurements of COP range of motion and velocity are potentially useful for monitoring changes in balance as an indicator of core stability during equine-assisted activities.

  6. Interaction of COP1 and UVR8 regulates UV-B-induced photomorphogenesis and stress acclimation in Arabidopsis

    PubMed Central

    Favory, Jean-Jacques; Stec, Agnieszka; Gruber, Henriette; Rizzini, Luca; Oravecz, Attila; Funk, Markus; Albert, Andreas; Cloix, Catherine; Jenkins, Gareth I; Oakeley, Edward J; Seidlitz, Harald K; Nagy, Ferenc; Ulm, Roman

    2009-01-01

    The ultraviolet-B (UV-B) portion of the solar radiation functions as an environmental signal for which plants have evolved specific and sensitive UV-B perception systems. The UV-B-specific UV RESPONSE LOCUS 8 (UVR8) and the multifunctional E3 ubiquitin ligase CONSTITUTIVELY PHOTOMORPHOGENIC 1 (COP1) are key regulators of the UV-B response. We show here that uvr8-null mutants are deficient in UV-B-induced photomorphogenesis and hypersensitive to UV-B stress, whereas overexpression of UVR8 results in enhanced UV-B photomorphogenesis, acclimation and tolerance to UV-B stress. By using sun simulators, we provide evidence at the physiological level that UV-B acclimation mediated by the UV-B-specific photoregulatory pathway is indeed required for survival in sunlight. At the molecular level, we demonstrate that the wild type but not the mutant UVR8 and COP1 proteins directly interact in a UV-B-dependent, rapid manner in planta. These data collectively suggest that UV-B-specific interaction of COP1 and UVR8 in the nucleus is a very early step in signalling and responsible for the plant's coordinated response to UV-B ensuring UV-B acclimation and protection in the natural environment. PMID:19165148

  7. Periplasmic response upon disruption of transmembrane Cu transport in Pseudomonas aeruginosa.

    PubMed

    Raimunda, Daniel; Padilla-Benavides, Teresita; Vogt, Stefan; Boutigny, Sylvain; Tomkinson, Kaleigh N; Finney, Lydia A; Argüello, José M

    2013-02-01

    Pseudomonas aeruginosa, an opportunistic pathogen, has two transmembrane Cu(+) transport ATPases, CopA1 and CopA2. Both proteins export cytoplasmic Cu(+) into the periplasm and mutation of either gene leads to attenuation of virulence. CopA1 is required for maintaining cytoplasmic copper levels, while CopA2 provides copper for cytochrome c oxidase assembly. We hypothesized that transported Cu(+) ions would be directed to their destination via specific periplasmic partners and disruption of transport should affect the periplasmic copper homeostasis. Supporting this, mutation of either ATPase gene led to large increments in periplasmic cuproprotein levels. Toward identifying the proteins participating in this cellular response the periplasmic metalloproteome was resolved in non-denaturing bidimensional gel electrophoresis, followed by X-ray fluorescence visualization and identification by mass-spectrometry. A single spot containing the electron shuttle protein azurin was responsible for the observed increments in cuproprotein contents. In agreement, lack of either Cu(+)-ATPase induced an increase in azu transcription. This is associated with an increase in the expression of anr and rpoS oxidative stress response regulators, rather than cueR, a copper sensing regulator. We propose that azurin overexpression and accumulation in the periplasm is part of the cellular response to cytoplasmic oxidative stress in P. aeruginosa.

  8. 30 CFR 285.638 - What must I do upon completion of my commercial operations as approved in my COP or FERC license?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... operations as approved in my COP or FERC license? 285.638 Section 285.638 Mineral Resources BUREAU OF OCEAN... Activities Under An Approved Cop § 285.638 What must I do upon completion of my commercial operations as approved in my COP or FERC license? (a) Upon completion of your approved activities under your COP, you...

  9. 30 CFR 585.635 - What must I do if I cease activities approved in my COP before the end of my commercial lease?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... in my COP before the end of my commercial lease? 585.635 Section 585.635 Mineral Resources BUREAU OF... Approved Cop § 585.635 What must I do if I cease activities approved in my COP before the end of my..., without an approved suspension, under your approved COP. If you cease commercial operations for an...

  10. 30 CFR 585.635 - What must I do if I cease activities approved in my COP before the end of my commercial lease?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... in my COP before the end of my commercial lease? 585.635 Section 585.635 Mineral Resources BUREAU OF... Approved Cop § 585.635 What must I do if I cease activities approved in my COP before the end of my..., without an approved suspension, under your approved COP. If you cease commercial operations for an...

  11. 30 CFR 585.635 - What must I do if I cease activities approved in my COP before the end of my commercial lease?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... in my COP before the end of my commercial lease? 585.635 Section 585.635 Mineral Resources BUREAU OF... Approved Cop § 585.635 What must I do if I cease activities approved in my COP before the end of my..., without an approved suspension, under your approved COP. If you cease commercial operations for an...

  12. New, Improved Version of the mCOP-PCR Screening System for Detection of Spinal Muscular Atrophy Gene (SMN1) Deletion.

    PubMed

    Shinohara, Masakazu; Ar Rochmah, Mawaddah; Nakanishi, Kenta; Harahap, Nur Imma Fatimah; Niba, Emma Tabe Eko; Saito, Toshio; Saito, Kayoko; Takeuchi, Atsuko; Bouike, Yoshihiro; Nishio, Hisahide

    2017-09-07

    Spinal muscular atrophy (SMA) is a frequent autosomal recessive disorder, characterized by lower motor neuron loss in the spinal cord. More than 95% of SMA patients show homozygous survival motor neuron 1 (SMN1) deletion. We previously developed a screening system for SMN1 deletion based on a modified competitive oligonucleotide priming-PCR (mCOP-PCR) technique. However, non-specific amplification products were observed with mCOP-PCR, which might lead to erroneous interpretation of the screening results. To establish an improved version of the mCOP-PCR screening system without non-specific amplification. DNA samples were assayed using a new version of the mCOP-PCR screening system. DNA samples had already been genotyped by PCR-restriction fragment length polymorphism (PCR-RFLP), showing the presence or absence of SMN1 exon 7. The new mCOP-PCR method contained a targeted pre-amplification step of the region, including an SMN1-specific nucleotide, prior to the mCOP-PCR step. mCOP-PCR products were electrophoresed on agarose gels. No non-specific amplification products were detected in electrophoresis gels with the new mCOP-PCR screening system. An additional targeted pre-amplification step eliminated non-specific amplification from mCOP-PCR screening.

  13. Effects of body lean and visual information on the equilibrium maintenance during stance.

    PubMed

    Duarte, Marcos; Zatsiorsky, Vladimir M

    2002-09-01

    Maintenance of equilibrium was tested in conditions when humans assume different leaning postures during upright standing. Subjects ( n=11) stood in 13 different body postures specified by visual center of pressure (COP) targets within their base of support (BOS). Different types of visual information were tested: continuous presentation of visual target, no vision after target presentation, and with simultaneous visual feedback of the COP. The following variables were used to describe the equilibrium maintenance: the mean of the COP position, the area of the ellipse covering the COP sway, and the resultant median frequency of the power spectral density of the COP displacement. The variability of the COP displacement, quantified by the COP area variable, increased when subjects occupied leaning postures, irrespective of the kind of visual information provided. This variability also increased when vision was removed in relation to when vision was present. Without vision, drifts in the COP data were observed which were larger for COP targets farther away from the neutral position. When COP feedback was given in addition to the visual target, the postural control system did not control stance better than in the condition with only visual information. These results indicate that the visual information is used by the postural control system at both short and long time scales.

  14. A phylogenomic analysis of the Actinomycetales mce operons

    PubMed Central

    Casali, Nicola; Riley, Lee W

    2007-01-01

    Background The genome of Mycobacterium tuberculosis harbors four copies of a cluster of genes termed mce operons. Despite extensive research that has demonstrated the importance of these operons on infection outcome, their physiological function remains obscure. Expanding databases of complete microbial genome sequences facilitate a comparative genomic approach that can provide valuable insight into the role of uncharacterized proteins. Results The M. tuberculosis mce loci each include two yrbE and six mce genes, which have homology to ABC transporter permeases and substrate-binding proteins, respectively. Operons with an identical structure were identified in all Mycobacterium species examined, as well as in five other Actinomycetales genera. Some of the Actinomycetales mce operons include an mkl gene, which encodes an ATPase resembling those of ABC uptake transporters. The phylogenetic profile of Mkl orthologs exactly matched that of the Mce and YrbE proteins. Through topology and motif analyses of YrbE homologs, we identified a region within the penultimate cytoplasmic loop that may serve as the site of interaction with the putative cognate Mkl ATPase. Homologs of the exported proteins encoded adjacent to the M. tuberculosis mce operons were detected in a conserved chromosomal location downstream of the majority of Actinomycetales operons. Operons containing linked mkl, yrbE and mce genes, resembling the classic organization of an ABC importer, were found to be common in Gram-negative bacteria and appear to be associated with changes in properties of the cell surface. Conclusion Evidence presented suggests that the mce operons of Actinomycetales species and related operons in Gram-negative bacteria encode a subfamily of ABC uptake transporters with a possible role in remodeling the cell envelope. PMID:17324287

  15. Toward High-Performance and Low-Cost Hydrogen Evolution Reaction Electrocatalysts: Nanostructuring Cobalt Phosphide (CoP) Particles on Carbon Fiber Paper.

    PubMed

    Yu, Shu Hearn; Chua, Daniel H C

    2018-05-02

    In this communication, we facily fabricated nanostructured CoP particles (150 to 200 nm) on carbon fiber paper (CFP) for hydrogen evolution reaction (HER) by a simple two-step process via a green route. In the first step, crystalline Co 3 O 4 nanocubes (150-200 nm) were loaded on CFP through a hydrothermal process at low temperature (120 °C). Interestingly, crystalline Co 3 O 4 nanocubes with a size 150-200 nm exhibited different growth mechanisms in contrast to the crystalline Co 3 O 4 nanocubes with a size <100 nm reported earlier. In the second step, these crystalline Co 3 O 4 nanocubes were converted to catalytically active CoP particles through chemical vapor deposition (CVD) phosphorization (denoted as CoP/CFP-H). Remarkably, CoP/CFP-H exhibited a low Tafel slope of 49.7 mV/dec and only required overpotentials of 128.1, 144.4, and 190.8 mV to drive geometric current densities of -10, -20, and -100 mA cm -2 , respectively. Besides, the CoP/CFP-H also demonstrated an excellent durability in an acidic environment under 2000 sweeps at a high scan rate (100 mV s -1 ) and a 24 h chronopotentiometry testing. For comparison, CoP was also fabricated through the electrodeposition method, followed by CVD phosphorization (denoted as CoP/CFP-E). It was found that the latter had exhibited inferior activity compared to CoP/CFP-H. The good performances of CoP/CFP-H are essentially due to the rational designs of electrode: (i) the applications of highly HER active CoP electrocatalyst, (ii) the intimate contact of nanostructured CoP on carbon fibers, and (iii) the large electrochemical surface area at electrocatalyst/electrolyte interface due to the large retaining of particles features after phosphorization. Notably, the intermediate Co 3 O 4 /CFP can serve as a platform to develop other cobalt-based functional materials.

  16. In Situ Space Gas Dynamic Measurements by the ROSINA Comet Pressure Sensor COPS on the Rosetta Spacecraft

    NASA Astrophysics Data System (ADS)

    Tzou, C. Y.; Altwegg, K.; Fiethe, B.; Gasc, S.; Rubin, M.

    2014-12-01

    Rosetta is part of the cornerstone missions executed by the European Space Agency. It is the first space mission to orbit and also land on a comet. Starting in August 2014 Rosetta will be able to carry out a close study of comet 67P/Churyumov-Gerasimenko. The Rosetta Orbiter Spectrometer for Ion and Neutral Analysis (ROSINA) is one of the core payloads on board of the Rosetta spacecraft [Balsiger et al, 2007]. ROSINA's main objective is to determine the major atmospheric and ionospheric composition in the coma and to investigate the gas dynamics around the comet. ROSINA consists of two mass spectrometers and a pressure sensor. The Comet Pressure Sensor (COPS) includes two gauges: the "nude gauge" measures total neutral density in the coma and the "ram gauge" measures the dynamic pressure of the cometary gas flux to obtain the bulk velocity of the neutral gas. The combination of these two gauges makes COPS capable to derive the gas dynamics at the location of the spacecraft. We performed laboratory gas dynamic measurements with the identical flight-spare instrument of COPS. Using the Calibration System for The Mass Spectrometer Instrument ROSINA (CASYMIR) we produce neutral gas beams to model cometary gas jets with velocities from thermal up to 2 km/s. We expect that COPS will be able to detect the faint and expanding atmosphere of comet 67P/Churyumov-Gerasimenko as early as August 2014 when the comet is still farther than 3 AU from the Sun. We will present the first ROSINA COPS observations of the gas dynamics around the comet together with the corresponding laboratory measurements required for the interpretation of these data. Reference: Balsiger, H. et al.: ROSINA-Rosetta Orbiter Spectrometer for Ion and Neutral Analysis, Space Science Reviews, Vol. 128, 745-801, 2007.

  17. Sequence and features of the tryptophan operon of Vibrio parahemolyticus.

    PubMed

    Crawford, I P; Han, C Y; Silverman, M

    1991-01-01

    The nucleotide sequence of the trp operon of the marine enteric bacterium Vibrio parahemolyticus is presented. The gene order E, G, D, C(F), B, A is identical to that of other enterics. The structural genes of the operon are preceded by a long leader region encoding a 41-residue peptide containing five tryptophan residues. The organization of the leader region suggests that transcription of the operon is subject to attenuation control. The promoter-operator region of the V. parahemolyticus trp operon is almost identical to the corresponding promoter-operator of E. coli. The similarities suggest that promoter strength and operator function are identical in the two species, and that transcription initiation is regulated by repression. The operon appears to lack the internal promoter within trpD that is common in terrestrial enteric species.

  18. Copper tolerance in Frankia sp. strain EuI1c involves surface binding and copper transport.

    PubMed

    Rehan, Medhat; Furnholm, Teal; Finethy, Ryan H; Chu, Feixia; El-Fadly, Gomaah; Tisa, Louis S

    2014-09-01

    Several Frankia strains have been shown to be copper-tolerant. The mechanism of their copper tolerance was investigated for Frankia sp. strain EuI1c. Copper binding was shown by binding studies. Unusual globular structures were observed on the surface of the bacterium. These globular structures were composed of aggregates containing many relatively smaller "leaf-like" structures. Scanning electron microscopy with energy-dispersive X-ray (SEM-EDAX) analysis of these structures indicated elevated copper and phosphate levels compared to the control cells. Fourier transform infrared spectroscopy (FTIR) analysis indicated an increase in extracellular phosphate on the cell surface of copper-stressed cells. Bioinformatics' analysis of the Frankia sp. strain EuI1c genome revealed five potential cop genes: copA, copZ, copC, copCD, and copD. Experiments with Frankia sp. strain EuI1c using qRT-PCR indicated an increase in messenger RNA (mRNA) levels of the five cop genes upon Cu(2+) stress. After 5 days of Cu(2+) stress, the copA, copZ, copC, copCD, and copD mRNA levels increased 25-, 8-, 18-, 18-, and 25-fold, respectively. The protein profile of Cu(2+)-stressed Frankia sp. strain EuI1c cells revealed the upregulation of a 36.7 kDa protein that was identified as FraEuI1c_1092 (sulfate-binding periplasmic transport protein). Homologues of this gene were only present in the genomes of the Cu(2+)-resistant Frankia strains (EuI1c, DC12, and CN3). These data indicate that copper tolerance by Frankia sp. strain EuI1c involved the binding of copper to the cell surface and transport proteins.

  19. 20 CFR 10.224 - What happens if OWCP finds that the employee is not entitled to COP after it has been paid?

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... is not entitled to COP after it has been paid? 10.224 Section 10.224 Employees' Benefits OFFICE OF... Termination of Cop § 10.224 What happens if OWCP finds that the employee is not entitled to COP after it has been paid? Where OWCP finds that the employee is not entitled to COP after it has been paid, the...

  20. 20 CFR 10.224 - What happens if OWCP finds that the employee is not entitled to COP after it has been paid?

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... is not entitled to COP after it has been paid? 10.224 Section 10.224 Employees' Benefits OFFICE OF... Termination of Cop § 10.224 What happens if OWCP finds that the employee is not entitled to COP after it has been paid? Where OWCP finds that the employee is not entitled to COP after it has been paid, the...

  1. 20 CFR 10.224 - What happens if OWCP finds that the employee is not entitled to COP after it has been paid?

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... is not entitled to COP after it has been paid? 10.224 Section 10.224 Employees' Benefits OFFICE OF... Termination of Cop § 10.224 What happens if OWCP finds that the employee is not entitled to COP after it has been paid? Where OWCP finds that the employee is not entitled to COP after it has been paid, the...

  2. 20 CFR 10.224 - What happens if OWCP finds that the employee is not entitled to COP after it has been paid?

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... is not entitled to COP after it has been paid? 10.224 Section 10.224 Employees' Benefits OFFICE OF... Termination of Cop § 10.224 What happens if OWCP finds that the employee is not entitled to COP after it has been paid? Where OWCP finds that the employee is not entitled to COP after it has been paid, the...

  3. 20 CFR 10.224 - What happens if OWCP finds that the employee is not entitled to COP after it has been paid?

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... is not entitled to COP after it has been paid? 10.224 Section 10.224 Employees' Benefits OFFICE OF... Termination of Cop § 10.224 What happens if OWCP finds that the employee is not entitled to COP after it has been paid? Where OWCP finds that the employee is not entitled to COP after it has been paid, the...

  4. High hydrostatic pressures in traumatic joints require elevated synovial capillary pressure probably associated with arteriolar vasodilatation.

    PubMed

    Ahlqvist, J; Harilainen, A; Aalto, K; Sarna, S; Lalla, M; Osterlund, K

    1994-11-01

    Three out of the four Starling pressures were determined at arthroscopy of traumatic effusions of the knee. The range of the joint fluid hydrostatic pressure Pjoint was 5-83 cmH2O (0.5-8.1 kPa, 4-61 mmHg), that of the colloid osmotic pressure difference COPplasma-COPjoint 0-21.7 cmH2O. In 11 of 15 cases the sum Pjoint+COP difference exceeded 32.6 cmH2O (3.19 kPa, 24 mmHg), a high estimate of average capillary pressure at the level of the heart. The number of 'exceeding' cases was 8/15 if only 80% of the COP difference was considered effective. Pjoint and the COP difference oppose filtration of fluid from plasma into joints, indicating that mean capillary pressure, the only Starling pressure not determined, was elevated unless the effusions were being resorbed back into the blood. The findings can be explained by tamponade compensated by arteriolar vasodilatation, suspected to be metabolically mediated.

  5. COP improvement of refrigerator/freezers, air-conditioners, and heat pumps using nonazeotropic refrigerant mixtures

    NASA Technical Reports Server (NTRS)

    Westra, Douglas G.

    1993-01-01

    With the February, 1992 announcement by President Bush to move the deadline for outlawing CFC (chloro-fluoro-carbon) refrigerants from the year 2000 to the year 1996, the refrigeration and air-conditioning industries have been accelerating their efforts to find alternative refrigerants. Many of the alternative refrigerants being evaluated require synthetic lubricants, are less efficient, and have toxicity problems. One option to developing new, alternative refrigerants is to combine existing non-CFC refrigerants to form a nonazeotropic mixture, with the concentration optimized for the given application so that system COP (Coefficient Of Performance) may be maintained or even improved. This paper will discuss the dilemma that industry is facing regarding CFC phase-out and the problems associated with CFC alternatives presently under development. A definition of nonazeotropic mixtures will be provided, and the characteristics and COP benefits of nonazeotropic refrigerant mixtures will be explained using thermodynamic principles. Limitations and disadvantages of nonazeotropic mixtures will be discussed, and example systems using such mixtures will be reviewed.

  6. δ-COP contains a helix C-terminal to its longin domain key to COPI dynamics and function

    PubMed Central

    Arakel, Eric C.; Richter, Kora P.; Clancy, Anne; Schwappach, Blanche

    2016-01-01

    Membrane recruitment of coatomer and formation of coat protein I (COPI)-coated vesicles is crucial to homeostasis in the early secretory pathway. The conformational dynamics of COPI during cargo capture and vesicle formation is incompletely understood. By scanning the length of δ-COP via functional complementation in yeast, we dissect the domains of the δ-COP subunit. We show that the μ-homology domain is dispensable for COPI function in the early secretory pathway, whereas the N-terminal longin domain is essential. We map a previously uncharacterized helix, C-terminal to the longin domain, that is specifically required for the retrieval of HDEL-bearing endoplasmic reticulum-luminal residents. It is positionally analogous to an unstructured linker that becomes helical and membrane-facing in the open form of the AP2 clathrin adaptor complex. Based on the amphipathic nature of the critical helix it may probe the membrane for lipid packing defects or mediate interaction with cargo and thus contribute to stabilizing membrane-associated coatomer. PMID:27298352

  7. Consultative Committee for Space Data Systems recommendation for space data system standards: Telecommand. Part 2.1: Command operation procedures

    NASA Technical Reports Server (NTRS)

    1991-01-01

    This recommendation contains the detailed specification of the logic required to carry out the Command Operations Procedures of the Transfer Layer. The Recommendation for Telecommand--Part 2, Data Routing Service contains the standard data structures and data communication procedures used by the intermediate telecommand system layers (the Transfer and Segmentation Layers). In particular, it contains a brief description of the Command Operations Procedures (COP) within the Transfer Layer. This recommendation contains the detailed definition of the COP's in the form of state tables, along with definitions of the terms used. It is assumed that the reader of this document is familiar with the data structures and terminology of part 2. In case of conflict between the description of the COP's in part 2 and in this recommendation, the definition in this recommendation will take precedence. In particular, this document supersedes section 4.3.3.1 through 4.3.3.4 of part 2.

  8. COP1, a negative regulator of photomorphogenesis, positively regulates plant disease resistance via double-stranded RNA binding proteins

    PubMed Central

    Lim, Gah-Hyun; Zhu, Shifeng; Clavel, Marion; Yu, Keshun; Navarre, Duroy; Kachroo, Aardra; Deragon, Jean-Marc

    2018-01-01

    The E3 ubiquitin ligase COP1 (Constitutive Photomorphogenesis 1) is a well known component of the light-mediated plant development that acts as a repressor of photomorphogenesis. Here we show that COP1 positively regulates defense against turnip crinkle virus (TCV) and avrRPM1 bacteria by contributing to stability of resistance (R) protein HRT and RPM1, respectively. HRT and RPM1 levels and thereby pathogen resistance is significantly reduced in the cop1 mutant background. Notably, the levels of at least two double-stranded RNA binding (DRB) proteins DRB1 and DRB4 are reduced in the cop1 mutant background suggesting that COP1 affects HRT stability via its effect on the DRB proteins. Indeed, a mutation in either drb1 or drb4 resulted in degradation of HRT. In contrast to COP1, a multi-subunit E3 ligase encoded by anaphase-promoting complex (APC) 10 negatively regulates DRB4 and TCV resistance but had no effect on DRB1 levels. We propose that COP1-mediated positive regulation of HRT is dependent on a balance between COP1 and negative regulators that target DRB1 and DRB4. PMID:29513740

  9. Center of Pressure Trajectory during Gait: A Comparison of Four Foot Positions

    PubMed Central

    Lugade, Vipul; Kaufman, Kenton

    2014-01-01

    Knowledge of the center of pressure (COP) trajectory during stance can elucidate possible foot pathology, provide comparative effectiveness of foot orthotics, and allow for appropriate calculation of balance control and joint kinetics during gait. Therefore, the goal of this study was to investigate the COP movement when walking at self-selected speeds with plantigrade, equinus, inverted, and everted foot positions. A total of 13 healthy subjects were asked to walk barefoot across an 8 meter walkway with embedded force plates. The COP was computed for each stance limb using the ground reaction forces and moments collected from three force plates. Results demonstrated that the COP excursion was 83% of the foot length and 27% of the foot width in the anterior-posterior and medial lateral directions for plantigrade walking, respectively. Regression equations explained 94% and 44% of the anterior-posterior and medial-lateral COP variability during plantigrade walking. While the range of motion and COP velocity was similar for inverted and everted walking, the COP remained on the lateral and medial aspects of the foot for these two walking conditions, respectively. A reduced anterior-posterior COP range of motion and velocity was demonstrated during equinus walking. Ankle joint motion in the frontal and sagittal planes supported this COP movement, with increased inversion and plantar flexion demonstrated during inverted and equinus conditions, respectively. Results from this study demonstrated the COP kinematics during simulated pathological gait conditions, with the COP trajectory providing an additional tool for the evaluation of patients with pathology. PMID:24447906

  10. Association Between Circulating Osteogenic Progenitor Cells and Disability and Frailty in Older Persons: The Nepean Osteoporosis and Frailty Study.

    PubMed

    Gunawardene, Piumali; Bermeo, Sandra; Vidal, Christopher; Al-Saedi, Ahmed; Chung, Philip; Boersma, Derek; Phu, Steven; Pokorski, Izabella; Suriyaarachchi, Pushpa; Demontiero, Oddom; Duque, Gustavo

    2016-09-01

    Circulating osteogenic progenitor (COP) cells are considered as surrogates of the mesenchymal repository in the body. In this study, we hypothesized that COP cells decrease with age and that lower levels of COP cells are associated with greater frailty and disability in older persons. Using well-established clinical criteria, we quantified physical performance and disability and stratified frailty in a random sample of community-dwelling individuals enrolled in the Nepean Osteoporosis and Frailty (NOF) Study (mean age 82.8; N = 77; 70% female; 27 nonfrail, 23 prefrail, and 27 frail). Percentage of COP cells was quantified by flow cytometry. Logistic regression models estimated the relationship between the percentage of COP cells and prevalent disability, poor physical performance, and frailty. We found that aging is associated with a significant decrease in COP cells (p < .001). Lower percentages of COP cells were associated with disability and poor physical performance (p < .001). Older adults with COP cells in the lower quartile were more likely to be frail (odds ratio 2.65, 95% confidence interval 2.72-3.15, p < .001). In conclusion, COP cells in the circulation decrease with age. Lower percentages of COP cells in late life are associated with prevalent frailty and disability. Further longitudinal studies are needed to understand COP cells as a risk stratifier, biomarker, or therapeutic target and to predict disability in frail older persons. © The Author 2015. Published by Oxford University Press on behalf of The Gerontological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  11. 77 FR 68149 - Agency Information Collection Activities; Proposed New Collection; Comments Requested: COPS...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-11-15

    ... Collection; Comments Requested: COPS Comparative Assessment of Cost Reduction by Agencies Survey ACTION: 60... Collection: Proposed new collection; comments requested. (2) Title of the Form/Collection: COPS Comparative... Office grants or cooperative agreements will be asked complete the COPS Comparative Assessment of Cost...

  12. 78 FR 2692 - Agency Information Collection Activities; Proposed New Collection; Comments Requested: COPS...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-01-14

    ... Collection; Comments Requested: COPS Comparative Assessment of Cost Reduction by Agencies Survey ACTION: 30...; comments requested. (2) Title of the Form/Collection: COPS Comparative Assessment of Cost Reduction by... will be asked complete the COPS Comparative Assessment of Cost Reduction Survey. The survey will be...

  13. 32 CFR 635.24 - Updating the COPS MPRS.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 32 National Defense 4 2014-07-01 2013-07-01 true Updating the COPS MPRS. 635.24 Section 635.24... CRIMINAL INVESTIGATIONS LAW ENFORCEMENT REPORTING Offense Reporting § 635.24 Updating the COPS MPRS... ensure that every founded offense is reported into the COPS MPRS. Timely and accurate reporting is...

  14. 32 CFR 635.24 - Updating the COPS MPRS.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 32 National Defense 4 2012-07-01 2011-07-01 true Updating the COPS MPRS. 635.24 Section 635.24... CRIMINAL INVESTIGATIONS LAW ENFORCEMENT REPORTING Offense Reporting § 635.24 Updating the COPS MPRS... ensure that every founded offense is reported into the COPS MPRS. Timely and accurate reporting is...

  15. 32 CFR 635.24 - Updating the COPS MPRS.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 32 National Defense 4 2013-07-01 2013-07-01 false Updating the COPS MPRS. 635.24 Section 635.24... CRIMINAL INVESTIGATIONS LAW ENFORCEMENT REPORTING Offense Reporting § 635.24 Updating the COPS MPRS... ensure that every founded offense is reported into the COPS MPRS. Timely and accurate reporting is...

  16. Multiple regulatory elements for the glpA operon encoding anaerobic glycerol-3-phosphate dehydrogenase and the glpD operon encoding aerobic glycerol-3-phosphate dehydrogenase in Escherichia coli: further characterization of respiratory control.

    PubMed

    Iuchi, S; Cole, S T; Lin, E C

    1990-01-01

    In Escherichia coli, sn-glycerol-3-phosphate can be oxidized by two different flavo-dehydrogenases, an anaerobic enzyme encoded by the glpACB operon and an aerobic enzyme encoded by the glpD operon. These two operons belong to the glp regulon specifying the utilization of glycerol, sn-glycerol-3-phosphate, and glycerophosphodiesters. In glpR mutant cells grown under conditions of low catabolite repression, the glpA operon is best expressed anaerobically with fumarate as the exogenous electron acceptor, whereas the glpD operon is best expressed aerobically. Increased anaerobic expression of glpA is dependent on the fnr product, a pleiotropic activator of genes involved in anaerobic respiration. In this study we found that the expression of a glpA1(Oxr) (oxygen-resistant) mutant operon, selected for increased aerobic expression, became less dependent on the FNR protein but more dependent on the cyclic AMP-catabolite gene activator protein complex mediating catabolite repression. Despite the increased aerobic expression of glpA1(Oxr), a twofold aerobic repressibility persisted. Moreover, anaerobic repression by nitrate respiration remained normal. Thus, there seems to exist a redox control apart from the FNR-mediated one. We also showed that the anaerobic repression of the glpD operon was fully relieved by mutations in either arcA (encoding a presumptive DNA recognition protein) or arcB (encoding a presumptive redox sensor protein). The arc system is known to mediate pleiotropic control of genes of aerobic function.

  17. LacR Is a Repressor of lacABCD and LacT Is an Activator of lacTFEG, Constituting the lac Gene Cluster in Streptococcus pneumoniae

    PubMed Central

    Afzal, Muhammad; Shafeeq, Sulman

    2014-01-01

    Comparison of the transcriptome of Streptococcus pneumoniae strain D39 grown in the presence of either lactose or galactose with that of the strain grown in the presence of glucose revealed the elevated expression of various genes and operons, including the lac gene cluster, which is organized into two operons, i.e., lac operon I (lacABCD) and lac operon II (lacTFEG). Deletion of the DeoR family transcriptional regulator lacR that is present downstream of the lac gene cluster revealed elevated expression of lac operon I even in the absence of lactose. This suggests a function of LacR as a transcriptional repressor of lac operon I, which encodes enzymes involved in the phosphorylated tagatose pathway in the absence of lactose or galactose. Deletion of lacR did not affect the expression of lac operon II, which encodes a lactose-specific phosphotransferase. This finding was further confirmed by β-galactosidase assays with PlacA-lacZ and PlacT-lacZ in the presence of either lactose or glucose as the sole carbon source in the medium. This suggests the involvement of another transcriptional regulator in the regulation of lac operon II, which is the BglG-family transcriptional antiterminator LacT. We demonstrate the role of LacT as a transcriptional activator of lac operon II in the presence of lactose and CcpA-independent regulation of the lac gene cluster in S. pneumoniae. PMID:24951784

  18. The Dickeya dadantii biofilm matrix consists of cellulose nanofibres, and is an emergent property dependent upon the type III secretion system and the cellulose synthesis operon.

    PubMed

    Jahn, Courtney E; Selimi, Dija A; Barak, Jeri D; Charkowski, Amy O

    2011-10-01

    Dickeya dadantii is a plant-pathogenic bacterium that produces cellulose-containing biofilms, called pellicles, at the air-liquid interface of liquid cultures. D. dadantii pellicle formation appears to be an emergent property dependent upon at least three gene clusters, including cellulose synthesis, type III secretion system (T3SS) and flagellar genes. The D. dadantii cellulose synthesis operon is homologous to that of Gluconacetobacter xylinus, which is used for industrial cellulose production, and the cellulose nanofibres produced by D. dadantii were similar in diameter and branching pattern to those produced by G. xylinus. Salmonella enterica, an enterobacterium closely related to D. dadantii, encodes a second type of cellulose synthesis operon, and it produced biofilm strands that differed in width and branching pattern from those of D. dadantii and G. xylinus. Unlike any previously described cellulose fibre, the D. dadantii cellulose nanofibres were decorated with bead-like structures. Mutation of the cellulose synthesis operon genes resulted in loss of cellulose synthesis and production of a cellulase-resistant biofilm. Mutation of other genes required for pellicle formation, including those encoding FliA (a sigma factor that regulates flagella production), HrpL (a sigma factor that regulates the T3SS), and AdrA, a GGDEF protein, affected both biofilm and cell morphology. Mutation of the cellulose synthase bcsA or of bcsC resulted in decreased accumulation of the T3SS-secreted protein HrpN.

  19. beta'-COP, a novel subunit of coatomer.

    PubMed Central

    Stenbeck, G; Harter, C; Brecht, A; Herrmann, D; Lottspeich, F; Orci, L; Wieland, F T

    1993-01-01

    Several lines of evidence favour the hypothesis that intracellular biosynthetic protein transport in eukaryotes is mediated by non-clathrin-coated vesicles (for a review see Rothman and Orci, 1992). The vesicles have been isolated and a set of their surface proteins has been characterized as coat proteins (COPs). These COPs exist in the cytosol as a preformed complex, the coatomer, which was prior to this study known to contain six subunits: four (alpha-, beta-, gamma- and delta-COP) with molecular weights between 160 and 58 kDa, and two additional proteins of approximately 36 and 20 kDa, epsilon- and xi-COP. Here we describe a novel subunit of the coatomer complex, beta'-COP. This subunit occurs in amounts stoichiometric to the established COPs both in the coatomer and in nonclathrin-coated vesicles and shows homology to the beta-subunits of trimeric G proteins. Images PMID:8334999

  20. Cryptogenic organizing pneumonia masquerading as lung carcinoma: A case report and review of the literature

    PubMed Central

    Huo, Ji-Ping; Liu, Cui; Jin, Bei-Bei; Duan, Feng-Xia; Mei, Sheng-Hui; Li, Xin-Gang; Zhao, Zhi-Gang

    2018-01-01

    Cryptogenic organizing pneumonia (COP) is a rare pulmonary disorder of unknown etiology. COP with hemoptysis as the primary presenting symptom has rarely been reported. The present study reported a case of COP that resembled lung carcinoma with hemoptysis as the only clinical symptom. The patient recovered well following thoracoscope surgery. A literature review of 119 COP cases between 1995 and 2015 was presented. Cough, fever and dyspnea were the most common clinical manifestations. The most common imaging manifestations were multiple or single consolidation, lung nodules, migratory sign, reversed halo sign, and multiple ground-glass opacity. A total of 3 cases exhibited COP accompanied by lung cancer. Glucocorticoids were effective for the majority of cases and invasive surgeries were implemented in most cases. The majority of cases recovered or relieved, and the prognosis of COP was relatively good. COP was easily confused with lung tumor and it is necessary to make differential diagnosis between COP and lung cancer. Invasive surgery should be avoided when possible to avoid or reduce patient trauma. PMID:29399056

  1. Diversity of sesquiterpene synthases in the basidiomycete Coprinus cinereus

    PubMed Central

    Agger, Sean; Lopez-Gallego, Fernando; Schmidt-Dannert, Claudia

    2009-01-01

    SUMMARY Fungi are a rich source of bioactive secondary metabolites and mushroom-forming fungi (Agaricomycetes) are especially known for the synthesis of numerous bioactive and often cytotoxic sesquiterpenoid secondary metabolites. Compared to the large number of sesquiterpene synthases identified in plants, less than a handful of unique sesquiterpene synthases have been described from fungi. Here we describe the functional characterization of six sesquiterpene synthases (Cop1 to Cop6) and two terpene oxidizing cytochrome P450 monooxygenases (Cox1 and Cox2) from Coprinus cinereus. The genes were cloned and, except for cop5, functionally expressed in Escherichia coli and/or Saccharomyces cerevisiae. Cop1 and Cop2 each synthesize germacrene A as the major product. Cop3 was identified as a α-muurolene synthase, an enzyme that has not been described previously, while Cop4 synthesizes δ-cadinene as its major product. Cop6 was originally annotated as a trichodiene synthase homolog, but instead was found to catalyze highly specific the synthesis of α-cuprenene. Co-expression of cop6 and the two monooxygenase genes next to it yields oxygenated α-cuprenene derivatives, including cuparophenol, suggesting that these genes encode the enzymes for the biosynthesis of antimicrobial quinone sesquiterpenoids (known as lagopodins) that were previously isolated from C. cinereus and other Coprinus species. PMID:19400802

  2. An assessment of efficient water heating options for an all-electric single family residence in a mixed-humid climate.

    PubMed

    Balke, Elizabeth C; Healy, William M; Ullah, Tania

    2016-12-01

    An evaluation of a variety of efficient water heating strategies for an all-electric single family home located in a mixed-humid climate is conducted using numerical modeling. The strategies considered include various combinations of solar thermal, heat pump, and electric resistance water heaters. The numerical model used in the study is first validated against a year of field data obtained on a dual-tank system with a solar thermal preheat tank feeding a heat pump water heater that serves as a backup. Modeling results show that this configuration is the most efficient of the systems studied over the course of a year, with a system coefficient of performance (COP sys ) of 2.87. The heat pump water heater alone results in a COP sys of 1.9, while the baseline resistance water heater has a COP sys of 0.95. Impacts on space conditioning are also investigated by considering the extra energy consumption required of the air source heat pump to remove or add heat from the conditioned space by the water heating system. A modified COP sys that incorporates the heat pump energy consumption shows a significant drop in efficiency for the dual tank configuration since the heat pump water heater draws the most heat from the space in the heating season while the high temperatures in the solar storage tank during the cooling season result in an added heat load to the space. Despite this degradation in the COP sys , the combination of the solar thermal preheat tank and the heat pump water heater is the most efficient option even when considering the impacts on space conditioning.

  3. Redox-responsive Covalent Organic Nanosheets from Viologens and Calix[4]arene for Iodine and Toxic Dye Capture.

    PubMed

    Skorjanc, Tina; Shetty, Dinesh; Sharma, Sudhir; Raya, Jesus; Traboulsi, Hassan; Han, Dong-Suk; Lalla, Jayesh; Newlon, Ryan; Jagannathan, Ramesh; Kirmizialtin, Serdal; Olsen, John-Carl; Trabolsi, Ali

    2018-04-17

    Owing to their chemical and thermal stabilities, high uptake capacities, and easy recyclability, covalent organic polymers (COPs) have shown promise as pollutant sponges. Here, we describe the use of diazo coupling to synthesize two cationic COPs, COP1++ and COP2++, that incorporate a viologen-based molecular switch and an organic macrocycle, calix[4]arene. Both COPs form nanosheets with height profiles of 6.00 and 8.00 nm, respectively, based on AFM measurements. The sheets remain morphologically intact upon one- or two-electron reductions of their viologen subunits. MD simulations of the dicationic COPs indicate that calix[4]arene adopts a partial cone conformation and that, in height, the individual 2D polymer layers are 5.48 Å in COP1++ and 5.65 Å in COP2++, which, together with the AFM measurements, suggests that the nanosheets are composed of 11 and 14 layers, respectively. The COPs, in either dicationic, radical cationic, or neutral form exhibit high affinity for iodine, reaching up to 200% mass increase when exposed to iodine vapor at 70 °C, which makes the materials among the best-performing nanosheets for iodine capture reported in the literature. In addition, the COPs effectively remove Congo red from solution in the pH range of 2 - 10, reaching nearly 100% removal within 15 minutes at acidic pH. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. Company observational post-marketing studies: drug risk assessment and drug research in special populations--a study-based analysis.

    PubMed

    Hasford, J; Lamprecht, T

    1998-01-01

    Company observational post-marketing studies (COPS) claim to provide essential data about drug risks and effectiveness in special populations not admitted to pre-approval clinical trials. Since COPS are often mainly regarded as a marketing activity, this study-based analysis tries to evaluate the scientific contributions of COPS. Thirty-five COPS were identified by hand-searching through medical journals, writing to pharmaceutical manufacturers and using MEDLINE. Fourteen COPS evaluated cardiovascular drugs, 9 evaluated NSAIDs and 12 evaluated various other indications. Thirty-five COPS listed effectiveness, 31 listed safety and 8 listed patient compliance as principal objectives. Not a single COPS included a control group. Seventeen of 21 evaluable COPS mentioned extensive exclusion criteria similar to those in clinical trials. Median observation time was 8 weeks, too short for chronic diseases and for adverse drug reactions with longer latency periods. One new adverse event was regarded. Global assessments of the outcomes by physicians dominated and were not based on objective clinical findings. None of the studies specified any details concerning the standardisation of observations or quality-control procedures. The current COPS scheme does not contribute significantly to our knowledge of drug safety and the effects in special populations. Despite serious criticism over the past 20 years, the poor quality of COPS compared with dramatic improvements of pre-approval trials - implies a need for detailed guidelines for non-experimental phase IV research, similar to the Good Clinical Practice-Guideline of the European Community.

  5. Reliability and feasibility of gait initiation centre-of-pressure excursions using a Wii® Balance Board in older adults at risk of falling.

    PubMed

    Lee, James; Webb, Graham; Shortland, Adam P; Edwards, Rebecca; Wilce, Charlotte; Jones, Gareth D

    2018-04-17

    Impairments in dynamic balance have a detrimental effect in older adults at risk of falls (OARF). Gait initiation (GI) is a challenging transitional movement. Centre of pressure (COP) excursions using force plates have been used to measure GI performance. The Nintendo Wii Balance Board (WBB) offers an alternative to a standard force plate for the measurement of CoP excursion. To determine the reliability of COP excursions using the WBB, and its feasibility within a 4-week strength and balance intervention (SBI) treating OARF. Ten OARF subjects attending SBI and ten young healthy adults, each performed three GI trials after 10 s of quiet stance from a standardised foot position (shoulder width) before walking forward 3 m to pick up an object. Averaged COP mediolateral (ML) and anteroposterior (AP) excursions (distance) and path-length time (GI-onset to first toe-off) were analysed. WBB ML (0.866) and AP COP excursion (0.895) reliability (ICC 3,1 ) was excellent, and COP path-length reliability was fair (0.517). Compared to OARF, healthy subjects presented with larger COP excursion in both directions and shorter COP path length. OARF subjects meaningfully improved their timed-up-and-go and ML COP excursion between weeks 1-4, while AP COP excursions, path length, and confidence-in-balance remained stable. COP path length and excursion directions probably measure different GI postural control attributes. Limitations in WBB accuracy and precision in transition tasks needs to be established before it can be used clinically to measure postural aspects of GI viably. The WBB could provide valuable clinical evaluation of balance function in OARF.

  6. Salt Stress and Ethylene Antagonistically Regulate Nucleocytoplasmic Partitioning of COP1 to Control Seed Germination1[OPEN

    PubMed Central

    Shi, Hui; Gu, Juntao; Dong, Jingao; Deng, Xing Wang

    2016-01-01

    Seed germination, a critical stage initiating the life cycle of a plant, is severely affected by salt stress. However, the underlying mechanism of salt inhibition of seed germination (SSG) is unclear. Here, we report that the Arabidopsis (Arabidopsis thaliana) CONSTITUTIVE PHOTOMORPHOGENESIS1 (COP1) counteracts SSG. Genetic assays provide evidence that SSG in loss of function of the COP1 mutant was stronger than this in the wild type. A GUS-COP1 fusion was constitutively localized to the nucleus in radicle cells. Salt treatment caused COP1 to be retained in the cytosol, but the addition of ethylene precursor 1-aminocyclopropane-1-carboxylate had the reverse effect on the translocation of COP1 to the nucleus, revealing that ethylene and salt exert opposite regulatory effects on the localization of COP1 in germinating seeds. However, loss of function of the ETHYLENE INSENSITIVE3 (EIN3) mutant impaired the ethylene-mediated rescue of the salt restriction of COP1 to the nucleus. Further research showed that the interaction between COP1 and LONG HYPOCOTYL5 (HY5) had a role in SSG. Correspondingly, SSG in loss of function of HY5 was suppressed. Biochemical detection showed that salt promoted the stabilization of HY5, whereas ethylene restricted its accumulation. Furthermore, salt treatment stimulated and ethylene suppressed transcription of ABA INSENSITIVE5 (ABI5), which was directly transcriptionally regulated by HY5. Together, our results reveal that salt stress and ethylene antagonistically regulate nucleocytoplasmic partitioning of COP1, thereby controlling Arabidopsis seed germination via the COP1-mediated down-regulation of HY5 and ABI5. These findings enhance our understanding of the stress response and have great potential for application in agricultural production. PMID:26850275

  7. Comparison of centre of gravity and centre of pressure patterns in the golf swing.

    PubMed

    Smith, Aimée C; Roberts, Jonathan R; Kong, Pui Wah; Forrester, Stephanie E

    2017-03-01

    Analysing the centre of pressure (COP) and centre of gravity (COG) could reveal stabilising strategies used by golfers throughout the golf swing. This study identified and compared golfers' COP and COG patterns throughout the golf swing in medial-lateral (ML) and anterior-posterior (AP) directions using principal component analysis (PCA) and examined their relationship to clubhead velocity. Three-dimensional marker trajectories were collected using Vicon motion analysis and force plate data from two Kistler force plates for 22 low-handicap golfers during drives. Golfers' COG and COP were expressed as a percentage distance between their feet. PCA was performed on COG and COP in ML and AP directions. Relationships between principal component (PC) scores were examined using Pearson correlation and regression analysis used to examine the relationship with clubhead velocity. ML COP movements varied in magnitude (PC1), rate of change and timing (PC2 and PC3). The COP and COG PC1 scores were strongly correlated in both directions (ML: r = 0.90, P < .05; AP: r = 0.81, P < .05). Clubhead velocity, explained by three PCs (74%), related to timing and rate of change in COP ML near downswing (PC2 and PC3) and timing of COG ML late backswing (PC2). The relationship between COP ML and COG ML PC1 scores identified extremes of COP and COG patterns in golfers and could indicate a golfer's dynamic balance. Golfers with earlier movement of COP to the front foot (PC2) and rate of change (PC3) patterns in ML COP, prior to the downswing, may be more likely to generate higher clubhead velocity.

  8. Cardiorespiratory optimal point during exercise testing as a predictor of all-cause mortality.

    PubMed

    Ramos, Plínio S; Araújo, Claudio Gil S

    2017-04-01

    The cardiorespiratory optimal point (COP) is a novel index, calculated as the minimum oxygen ventilatory equivalent (VE/VO 2 ) obtained during cardiopulmonary exercise testing (CPET). In this study we demonstrate the prognostic value of COP both independently and in combination with maximum oxygen consumption (VO 2 max) in community-dwelling adults. Maximal cycle ergometer CPET was performed in 3331 adults (66% men) aged 40-85 years, healthy (18%) or with chronic disease (81%). COP cut-off values of <22, 22-30, and >30 were selected based on the log-rank test. Risk discrimination was assessed using COP as an independent predictor and combined with VO 2 max. Median follow-up was 6.4 years (7.1% mortality). Subjects with COP >30 demonstrated increased mortality compared to those with COP <22 (hazard ratio [HR] 6.86, 95% confidence interval [CI] 3.69-12.75, p<0.001). Multivariate analysis including gender, age, body mass index, and the forced expiratory volume in 1 s/vital capacity ratio showed adjusted HR for COP >30 of 3.72 (95% CI 1.98-6.98; p<0.001) and for COP 22-30 of 2.15 (95% CI 1.15-4.03, p<0.001). Combining COP and VO 2 max data further enhanced risk discrimination. COP >30, either independently or in combination with low VO 2 max, is a good predictor of all-cause mortality in community-dwelling adults (healthy or with chronic disease). COP is a submaximal prognostic index that is simple to obtain and adds to CPET assessment, especially for adults unable or unwilling to achieve maximal exercise. Copyright © 2017 Sociedade Portuguesa de Cardiologia. Publicado por Elsevier España, S.L.U. All rights reserved.

  9. Age-related differences in quality of standing balance using a composite score.

    PubMed

    Pasma, Jantsje H; Bijlsma, Astrid Y; van der Bij, Mark D W; Arendzen, J Hans; Meskers, Carel G M; Maier, Andrea B

    2014-01-01

    Age-related differences in standing balance are not detected by testing the ability to maintain balance. Quality of standing balance might be more sensitive to detect age-related differences. To study age-related differences in quality of standing balance, center of pressure (CoP) movement was evaluated using a wide range of CoP parameters in several standing conditions in healthy young and old participants. In 35 healthy young (18-30 years) and 75 healthy old (70-80 years) participants, CoP movement was assessed in eight standing conditions on a force plate, including side-by-side, one-leg, semi-tandem and tandem stance, both with eyes open and eyes closed. Direction-specific CoP composite scores were calculated from standardized single CoP parameters (mean amplitude, amplitude variability, mean velocity, velocity variability and range) in anterior-posterior (AP) and medial-lateral (ML) direction. Linear regression analysis was used to detect age-related differences in single CoP parameters and composite scores - adjusted for gender, height and weight. Overall, single CoP parameters were higher in old compared to young participants, but no single CoP parameter consistently demonstrated the largest effect size for all standing conditions. Age-related differences were demonstrated for CoP composite scores in AP direction (tandem eyes open; semi-tandem eyes closed; p < 0.001). CoP composite scores in ML direction were consistently higher for all standing conditions in old compared to young participants (p < 0.001). CoP composite scores in ML direction were the most consistent parameters to detect age-related differences in quality of standing balance in healthy participants and might be of clinical value to detect subtle changes in quality of standing balance. © 2014 S. Karger AG, Basel

  10. Salt Stress and Ethylene Antagonistically Regulate Nucleocytoplasmic Partitioning of COP1 to Control Seed Germination.

    PubMed

    Yu, Yanwen; Wang, Juan; Shi, Hui; Gu, Juntao; Dong, Jingao; Deng, Xing Wang; Huang, Rongfeng

    2016-04-01

    Seed germination, a critical stage initiating the life cycle of a plant, is severely affected by salt stress. However, the underlying mechanism of salt inhibition of seed germination (SSG) is unclear. Here, we report that the Arabidopsis (Arabidopsis thaliana) CONSTITUTIVE PHOTOMORPHOGENESIS1 (COP1) counteracts SSG Genetic assays provide evidence that SSG in loss of function of the COP1 mutant was stronger than this in the wild type. A GUS-COP1 fusion was constitutively localized to the nucleus in radicle cells. Salt treatment caused COP1 to be retained in the cytosol, but the addition of ethylene precursor 1-aminocyclopropane-1-carboxylate had the reverse effect on the translocation of COP1 to the nucleus, revealing that ethylene and salt exert opposite regulatory effects on the localization of COP1 in germinating seeds. However, loss of function of the ETHYLENE INSENSITIVE3 (EIN3) mutant impaired the ethylene-mediated rescue of the salt restriction of COP1 to the nucleus. Further research showed that the interaction between COP1 and LONG HYPOCOTYL5 (HY5) had a role in SSG Correspondingly, SSG in loss of function of HY5 was suppressed. Biochemical detection showed that salt promoted the stabilization of HY5, whereas ethylene restricted its accumulation. Furthermore, salt treatment stimulated and ethylene suppressed transcription of ABA INSENSITIVE5 (ABI5), which was directly transcriptionally regulated by HY5. Together, our results reveal that salt stress and ethylene antagonistically regulate nucleocytoplasmic partitioning of COP1, thereby controlling Arabidopsis seed germination via the COP1-mediated down-regulation of HY5 and ABI5. These findings enhance our understanding of the stress response and have great potential for application in agricultural production. © 2016 American Society of Plant Biologists. All Rights Reserved.

  11. Cloning and Molecular Analysis of a Mannitol Operon of Phosphoenolpyruvate-dependent Phosphotransferase (PTS) type From Vibrio cholerae O395

    PubMed Central

    Kumar, Sanath; Smith, Kenneth P.; Floyd, Jody L.; Varela, Manuel F.

    2010-01-01

    A putative mannitol operon of the phosphoenolpyruvate phosphotransferase (PTS) type was cloned from Vibrio cholerae O395 and its activity studied in Escherichia coli. The 3.9 kb operon comprising of three genes is organized as mtlADR. Based on the sequence analysis, these were identified as genes encoding a putative mannitol-specific enzyme IICBA (EIIMtl) component (MtlA), a mannitol-1-phosphate dehydrogenase (MtlD) and a mannitol operon repressor (MtlR). The transport of [3H]mannitol by the cloned mannitol operon in E. coli was 13.8±1.4 nmol/min/mg protein. The insertional inactivation of EIIMtl abolished mannitol and sorbitol transport in V. cholerae O395. Comparison of the mannitol utilization apparatus of V. cholerae with those of Gram-negative and Gram positive bacteria suggests highly conserved nature of the system. MtlA and MtlD exhibit 75% similarity with corresponding sequences of E. coli mannitol operon genes, while MtlR has 63% similarity with MtlR of E. coli. The cloning of V. cholerae mannitol utilization system in an E. coli background will help in elucidating the functional properties of this operon. PMID:21184218

  12. Center of Pressure Motion After Calf Vibration Is More Random in Fallers Than Non-fallers: Prospective Study of Older Individuals

    PubMed Central

    van den Hoorn, Wolbert; Kerr, Graham K.; van Dieën, Jaap H.; Hodges, Paul W.

    2018-01-01

    Aging is associated with changes in balance control and elderly take longer to adapt to changing sensory conditions, which may increase falls risk. Low amplitude calf muscle vibration stimulates local sensory afferents/receptors and affects sense of upright when applied in stance. It has been used to assess the extent the nervous system relies on calf muscle somatosensory information and to rapidly change/perturb part of the somatosensory information causing balance unsteadiness by addition and removal of the vibratory stimulus. This study assessed the effect of addition and removal of calf vibration on balance control (in the absence of vision) in elderly individuals (>65 years, n = 99) who did (n = 41) or did not prospectively report falls (n = 58), and in a group of young individuals (18–25 years, n = 23). Participants stood barefoot and blindfolded on a force plate for 135 s. Vibrators (60 Hz, 1 mm) attached bilaterally over the triceps surae muscles were activated twice for 15 s; after 15 and 75 s (45 s for recovery). Balance measures were applied in a windowed (15 s epoch) manner to compare center-of-pressure (CoP) motion before, during and after removal of calf vibration between groups. In each epoch, CoP motion was quantified using linear measures, and non-linear measures to assess temporal structure of CoP motion [using recurrence quantification analysis (RQA) and detrended fluctuation analysis]. Mean CoP displacement during and after vibration did not differ between groups, which suggests that calf proprioception and/or weighting assigned by the nervous system to calf proprioception was similar for the young and both groups of older individuals. Overall, compared to the elderly, CoP motion of young was more predictable and persistent. Balance measures were not different between fallers and non-fallers before and during vibration. However, non-linear aspects of CoP motion of fallers and non-fallers differed after removal of vibration, when dynamic re-weighting is required. During this period fallers exhibited more random CoP motion, which could result from a reduced ability to control balance and/or a reduced ability to dynamically reweight proprioceptive information. These results show that non-linear measures of balance provide evidence for deficits in balance control in people who go on to fall in the following 12 months. PMID:29632494

  13. Structure Identification Using the US EPA's CompTox Chemistry Dashboard (CompTox CoP)

    EPA Science Inventory

    Community of practice webinar presentation on the Identification of unknowns in non-targeted analyses (NTA) requires the integration of complementary data types to generate a confident consensus structure.

  14. Water-filled training tubes increase core muscle activation and somatosensory control of balance during squat.

    PubMed

    Ditroilo, Massimiliano; O'Sullivan, Rory; Harnan, Brian; Crossey, Aislinn; Gillmor, Beth; Dardis, William; Grainger, Adam

    2018-09-01

    This study examined trunk muscle activation, balance and proprioception while squatting with a water-filled training tube (WT) and a traditional barbell (BB), with either closed (CE) or open eyes (OE). Eighteen male elite Gaelic footballers performed an isometric squat under the following conditions: BB-OE, BB-CE, WT-OE and WT-CE. The activity of rectus abdominis (RA), external oblique (EO) and multifidus (MF) was measured using electromyography, along with sway of the centre of pressure (CoP) using a force platform. Only the EO and the MF muscles exhibited an increased activity with WT (p < 0.01). In the medio-lateral direction both the velocity and range of the CoP increased significantly with WT (p < 0.01). Interestingly, the range of the CoP for the WT-CE condition was significantly lower than WT-OE (p < 0.05, d = 0.44), whilst the velocity of the CoP was marginally reduced (d = 0.29). WT elicited a greater level core muscle activation and created a greater challenge to postural stability when compared to a BB. It appears that WT does not benefit from vision but emphasises the somatosensory control of balance. The use of WT may be beneficial in those sports requiring development of somatosensory/proprioceptive contribution to balance control.

  15. Visual Reliance for Balance Control in Older Adults Persists When Visual Information Is Disrupted by Artificial Feedback Delays

    PubMed Central

    Balasubramaniam, Ramesh

    2014-01-01

    Sensory information from our eyes, skin and muscles helps guide and correct balance. Less appreciated, however, is that delays in the transmission of sensory information between our eyes, limbs and central nervous system can exceed several 10s of milliseconds. Investigating how these time-delayed sensory signals influence balance control is central to understanding the postural system. Here, we investigate how delayed visual feedback and cognitive performance influence postural control in healthy young and older adults. The task required that participants position their center of pressure (COP) in a fixed target as accurately as possible without visual feedback about their COP location (eyes-open balance), or with artificial time delays imposed on visual COP feedback. On selected trials, the participants also performed a silent arithmetic task (cognitive dual task). We separated COP time series into distinct frequency components using low and high-pass filtering routines. Visual feedback delays affected low frequency postural corrections in young and older adults, with larger increases in postural sway noted for the group of older adults. In comparison, cognitive performance reduced the variability of rapid center of pressure displacements in young adults, but did not alter postural sway in the group of older adults. Our results demonstrate that older adults prioritize vision to control posture. This visual reliance persists even when feedback about the task is delayed by several hundreds of milliseconds. PMID:24614576

  16. [Medical treatment for the management of painful endometriosis without infertility: CNGOF-HAS Endometriosis Guidelines].

    PubMed

    Sauvan, M; Chabbert-Buffet, N; Canis, M; Collinet, P; Fritel, X; Geoffron, S; Legendre, G; Wattier, J-M; Fernandez, H

    2018-03-01

    To provide clinical practice guidelines for the management of painful endometriosis in women without infertility. Systematic review of the literature literature since 2006, level of evidence rating, external proofreading and grading of the recommendation grade by an expert group according to HAS methodology. Combined hormonal contraceptives (COP) and the levonorgestrel-releasing intra-uterin system (LNG-IUS) are recommended as first-line hormonal therapies for the treatment of painful endometriosis (grade B). Second-line therapy relies on oral desogestrel microprogestative, etonogestrel-releasing implant, GnRH analogs (GnRHa) and dienogest (grade C). It is recommended to use add-back therapy containing estrogen in association with GnRHa (grade B). After endometriosis surgery, hormonal treatment relying on COP or LNG-IUS is recommended to prevent pain recurrence (grade B). COP is recommended to reduce the risk of endometrioma recurrence after surgery (grade B) but the prescription of GnRHa is not recommended (grade C). Continuous COP is recommended in case of dysmenorrhea (grade B). GnRHa is not recommended as first line endometriosis treatment for adolescent girl because of the risk of bone demineralization (grade B). The management of endometriosis-induced chronic pain requires an interdisciplinary evaluation. Physical therapies improving the quality of life such as yoga, relaxation or osteopathy can be proposed (expert agreement). Promising medical alternatives are currently under preclinical and clinical evaluation. Copyright © 2018 Elsevier Masson SAS. All rights reserved.

  17. Tactical Mission Command (TMC)

    DTIC Science & Technology

    2016-03-01

    capabilities to Army commanders and their staffs, consisting primarily of a user-customizable Common Operating Picture ( COP ) enabled with real-time... COP viewer and data management capability. It is a collaborative, visualization and planning application that also provides a common map display... COP ): Display the COP consisting of the following:1 Friendly forces determined by the commander including subordinate and supporting units at

  18. IRR (Inter-Rater Reliability) of a COP (Classroom Observation Protocol)--A Critical Appraisal

    ERIC Educational Resources Information Center

    Rui, Ning; Feldman, Jill M.

    2012-01-01

    Notwithstanding broad utility of COPs (classroom observation protocols), there has been limited documentation of the psychometric properties of even the most popular COPs. This study attempted to fill this void by closely examining the item and domain-level IRR (inter-rater reliability) of a COP that was used in a federally funded striving readers…

  19. Quantitative approaches to the study of bistability in the lac operon of Escherichia coli.

    PubMed

    Santillán, Moisés; Mackey, Michael C

    2008-08-06

    In this paper, the history and importance of the lac operon in the development of molecular and systems biology are briefly reviewed. We start by presenting a description of the regulatory mechanisms in this operon, taking into account the most recent discoveries. Then we offer a survey of the history of the lac operon, including the discovery of its main elements and the subsequent influence on the development of molecular and systems biology. Next the bistable behaviour of the operon is discussed, both with respect to its discovery and its molecular origin. A review of the literature in which this bistable phenomenon has been studied from a mathematical modelling viewpoint is then given. We conclude with some brief remarks.

  20. A mutation in a new gene bglJ, activates the bgl operon in Escherichia coli K-12

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

    Giel, M.; Desnoyer, M.; Lopilato, J.

    1996-06-01

    A new mutation , bglJ4, has been characterized that results in the expression of the silent bgl operon. The bgl operon encodes proteins necessary for the transport and utilization of the aromatic {beta}-glucosides arbutin and salicin. A variety of mutations activate the operon and result in a Bgl{sup +} phenotype. Activating mutations are located upstream of the bgl promoter and in genes located elsewhere on the chromosome. Mutations outside of the bgl operon occur in the genes encoding DNA gyrase and in the gene encoding the nucleoid associated protein H-NS. The mutation described here, bglJ4, has been mapped to amore » new locus at min 99 on the Escherichia coli K-12 genetic map. The putative protein encoded by the bglJ gene has homology to a family of transcriptional activators. Evidence is presented that increased expression of the bglJ product is needed for activation of the bgl operon. 56 refs., 3 figs., 3 tabs.« less

  1. Evidence for a common gene pool and frequent recombinational exchange of the tbpBA operon in Mannheimia haemolytica, Mannheimia glucosida and Bibersteinia trehalosi

    PubMed Central

    Lee, Inkyoung; Davies, Robert L.

    2012-01-01

    SUMMARY The tbpBA operon was sequenced in 42 representative isolates of Mannheimia haemolytica (32), Mannheimia glucosida (6) and Bibersteinia trehalosi (4). A total of 27 tbpB and 20 tbpA alleles were identified whilst the tbpBA operon was represented by 28 unique alleles that could be assigned to seven classes. There were 1566 (34.8% variation) polymorphic nucleotide sites and 482 (32.1% variation) variable inferred amino acid positions among the 42 tbpBA sequences. The tbpBA operons of serotype A2 M. haemolytica isolates are, with one exception, substantially more diverse than those of the other M. haemolytica serotypes and most likely have a different ancestral origin. The tbpBA phylogeny has been severely disrupted by numerous small- and large-scale intragenic recombination events. In addition, assortative (entire gene) recombination events, involving either the entire tbpBA operon or the individual tbpB and tbpA genes, have played a major role in shaping tbpBA structure and it’s distribution in the three species. Our findings indicate that a common gene pool exists for tbpBA in M. haemolytica, M. glucosida and B. trehalosi. In particular, B. trehalosi, M. glucosida and ovine M. haemolytica isolates share a large portion of the tbpA gene and this probably reflects selection for a conserved TbpA protein that provides effective iron-uptake in sheep. Bovine and ovine serotype A2 lineages have very different tbpBA alleles. Bovine-like tbpBA alleles have been partially, or completely, replaced by ovine-like tbpBA alleles in ovine serotype A2 isolates suggesting that different transferrin receptors are required by serotype A2 isolates for optimum iron uptake in cattle and sheep. Conversely, the tbpBA alleles of bovine-pathogenic serotype A1 and A6 isolates are very similar to those of closely related ovine isolates suggesting a recent and common evolutionary origin. PMID:20884693

  2. 20 CFR 10.222 - When may an employer terminate COP which has already begun?

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 20 Employees' Benefits 1 2011-04-01 2011-04-01 false When may an employer terminate COP which has...' COMPENSATION ACT, AS AMENDED Continuation of Pay Controversion and Termination of Cop § 10.222 When may an employer terminate COP which has already begun? (a) Where the employer has continued the pay of the...

  3. 20 CFR 10.222 - When may an employer terminate COP which has already begun?

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 20 Employees' Benefits 1 2014-04-01 2012-04-01 true When may an employer terminate COP which has...' COMPENSATION ACT, AS AMENDED Continuation of Pay Controversion and Termination of Cop § 10.222 When may an employer terminate COP which has already begun? (a) Where the employer has continued the pay of the...

  4. 20 CFR 10.222 - When may an employer terminate COP which has already begun?

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 20 Employees' Benefits 1 2012-04-01 2012-04-01 false When may an employer terminate COP which has...' COMPENSATION ACT, AS AMENDED Continuation of Pay Controversion and Termination of Cop § 10.222 When may an employer terminate COP which has already begun? (a) Where the employer has continued the pay of the...

  5. 30 CFR 585.658 - Can my cable or pipeline construction deviate from my approved COP or GAP?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... from my approved COP or GAP? 585.658 Section 585.658 Mineral Resources BUREAU OF OCEAN ENERGY... Can my cable or pipeline construction deviate from my approved COP or GAP? (a) You must make every... COP or GAP. (c) If, after construction, it is determined that a deviation from the approved plan has...

  6. Communicating Knowing through Communities of Practice: Exploring Internal Communicative Processes and Differences among CoPs

    ERIC Educational Resources Information Center

    Iverson, Joel O.; McPhee, Robert D.

    2008-01-01

    Knowing is an enacted, communicated process that is difficult to observe, let alone manage, in organizations. Communities of practice (CoPs) offer a productive solution for improving knowledge and knowledge management, but the communicative processes that enact CoPs have not been explored, leaving CoPs as an organizational black box. This research…

  7. 20 CFR 10.222 - When may an employer terminate COP which has already begun?

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 20 Employees' Benefits 1 2010-04-01 2010-04-01 false When may an employer terminate COP which has...' COMPENSATION ACT, AS AMENDED Continuation of Pay Controversion and Termination of Cop § 10.222 When may an employer terminate COP which has already begun? (a) Where the employer has continued the pay of the...

  8. 20 CFR 10.206 - May an employee who uses leave after an injury later decide to use COP instead?

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... later decide to use COP instead? 10.206 Section 10.206 Employees' Benefits OFFICE OF WORKERS... THE FEDERAL EMPLOYEES' COMPENSATION ACT, AS AMENDED Continuation of Pay Eligibility for Cop § 10.206 May an employee who uses leave after an injury later decide to use COP instead? On Form CA-1, an...

  9. 20 CFR 10.206 - May an employee who uses leave after an injury later decide to use COP instead?

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... later decide to use COP instead? 10.206 Section 10.206 Employees' Benefits OFFICE OF WORKERS... THE FEDERAL EMPLOYEES' COMPENSATION ACT, AS AMENDED Continuation of Pay Eligibility for Cop § 10.206 May an employee who uses leave after an injury later decide to use COP instead? On Form CA-1, an...

  10. 20 CFR 10.222 - When may an employer terminate COP which has already begun?

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 20 Employees' Benefits 1 2013-04-01 2012-04-01 true When may an employer terminate COP which has...' COMPENSATION ACT, AS AMENDED Continuation of Pay Controversion and Termination of Cop § 10.222 When may an employer terminate COP which has already begun? (a) Where the employer has continued the pay of the...

  11. 30 CFR 585.658 - Can my cable or pipeline construction deviate from my approved COP or GAP?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... from my approved COP or GAP? 585.658 Section 585.658 Mineral Resources BUREAU OF OCEAN ENERGY... Can my cable or pipeline construction deviate from my approved COP or GAP? (a) You must make every... COP or GAP. (c) If, after construction, it is determined that a deviation from the approved plan has...

  12. 30 CFR 585.658 - Can my cable or pipeline construction deviate from my approved COP or GAP?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... from my approved COP or GAP? 585.658 Section 585.658 Mineral Resources BUREAU OF OCEAN ENERGY... Can my cable or pipeline construction deviate from my approved COP or GAP? (a) You must make every... COP or GAP. (c) If, after construction, it is determined that a deviation from the approved plan has...

  13. The Importance of CoP in Transforming New Learning Communities into Experienced Ones in EFL Classrooms

    ERIC Educational Resources Information Center

    Nagao, Akiko

    2017-01-01

    Since the Communities of Practice (CoP) concept has been adopted in various learning environments, visualizing its development in English as a foreign language (EFL) classrooms is complicated. Thus, based on the CoP concept, this study investigates the changes in learners' degrees of participation and CoP elements in EFL writing/reading classes…

  14. Preparation and characterization of aminoethyl hydroxypropyl starch modified with collagen peptide.

    PubMed

    Wen, Huigao; Hu, Jin; Ge, Hongyu; Zou, Shengqiong; Xiao, Yao; Li, Ya; Feng, Han; Fan, Lihong

    2017-08-01

    The preparation of aminoethyl hydroxypropyl starch collagen peptide (AEHPS-COP) was via an enzyme-catalyzed reaction between amino groups in aminoethyl hydroxypropyl starch (AEHPS) and γ-carboxamide groups in collagen peptide (COP) by using microbial transglutaminase (MTGase) as biocatalyst. As an intermediate reactant, AEHPS was synthesized from hydroxypropyl starch (HPS) and 2-chloroethylamine hydrochloride (CEH). The chemical structures of HPS, AEHPS and AEHPS-COP were characterized by Fourier transform infrared spectroscopy (FT-IR) and 13 C nuclear magnetic resonance ( 13 C NMR). The reaction conditions that influenced the degree of substitution (DS) of AEHPS-COP were optimized, which included the reaction temperature, the reaction time, the mass ratio of collagen peptide to aminoethyl hydroxypropyl starch and the pH value. In addition, in vitro antioxidant activities of AEHPS-COP were evaluated through the scavenging activity of hydroxyl and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical. Furthermore, the methylthiazol tetrazolium (MTT) assay was applied to investigate the cell viability of AEHPS-COP. The results indicated that the AEHPS-COP exhibited better cell viability to L929 mouse fibroblast cells. Therefore, the AEHPS-COP showed a promising potential application in cosmetic, biomedical and pharmaceutical fields. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. A Novel Method for Accurate Operon Predictions in All SequencedProkaryotes

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

    Price, Morgan N.; Huang, Katherine H.; Alm, Eric J.

    2004-12-01

    We combine comparative genomic measures and the distance separating adjacent genes to predict operons in 124 completely sequenced prokaryotic genomes. Our method automatically tailors itself to each genome using sequence information alone, and thus can be applied to any prokaryote. For Escherichia coli K12 and Bacillus subtilis, our method is 85 and 83% accurate, respectively, which is similar to the accuracy of methods that use the same features but are trained on experimentally characterized transcripts. In Halobacterium NRC-1 and in Helicobacterpylori, our method correctly infers that genes in operons are separated by shorter distances than they are in E.coli, andmore » its predictions using distance alone are more accurate than distance-only predictions trained on a database of E.coli transcripts. We use microarray data from sixphylogenetically diverse prokaryotes to show that combining intergenic distance with comparative genomic measures further improves accuracy and that our method is broadly effective. Finally, we survey operon structure across 124 genomes, and find several surprises: H.pylori has many operons, contrary to previous reports; Bacillus anthracis has an unusual number of pseudogenes within conserved operons; and Synechocystis PCC6803 has many operons even though it has unusually wide spacings between conserved adjacent genes.« less

  16. Validating and calibrating the Nintendo Wii balance board to derive reliable center of pressure measures.

    PubMed

    Leach, Julia M; Mancini, Martina; Peterka, Robert J; Hayes, Tamara L; Horak, Fay B

    2014-09-29

    The Nintendo Wii balance board (WBB) has generated significant interest in its application as a postural control measurement device in both the clinical and (basic, clinical, and rehabilitation) research domains. Although the WBB has been proposed as an alternative to the "gold standard" laboratory-grade force plate, additional research is necessary before the WBB can be considered a valid and reliable center of pressure (CoP) measurement device. In this study, we used the WBB and a laboratory-grade AMTI force plate (AFP) to simultaneously measure the CoP displacement of a controlled dynamic load, which has not been done before. A one-dimensional inverted pendulum was displaced at several different displacement angles and load heights to simulate a variety of postural sway amplitudes and frequencies (<1 Hz). Twelve WBBs were tested to address the issue of inter-device variability. There was a significant effect of sway amplitude, frequency, and direction on the WBB's CoP measurement error, with an increase in error as both sway amplitude and frequency increased and a significantly greater error in the mediolateral (ML) (compared to the anteroposterior (AP)) sway direction. There was no difference in error across the 12 WBB's, supporting low inter-device variability. A linear calibration procedure was then implemented to correct the WBB's CoP signals and reduce measurement error. There was a significant effect of calibration on the WBB's CoP signal accuracy, with a significant reduction in CoP measurement error (quantified by root-mean-squared error) from 2-6 mm (before calibration) to 0.5-2 mm (after calibration). WBB-based CoP signal calibration also significantly reduced the percent error in derived (time-domain) CoP sway measures, from -10.5% (before calibration) to -0.05% (after calibration) (percent errors averaged across all sway measures and in both sway directions). In this study, we characterized the WBB's CoP measurement error under controlled, dynamic conditions and implemented a linear calibration procedure for WBB CoP signals that is recommended to reduce CoP measurement error and provide more reliable estimates of time-domain CoP measures. Despite our promising results, additional work is necessary to understand how our findings translate to the clinical and rehabilitation research domains. Once the WBB's CoP measurement error is fully characterized in human postural sway (which differs from our simulated postural sway in both amplitude and frequency content), it may be used to measure CoP displacement in situations where lower accuracy and precision is acceptable.

  17. Validating and Calibrating the Nintendo Wii Balance Board to Derive Reliable Center of Pressure Measures

    PubMed Central

    Leach, Julia M.; Mancini, Martina; Peterka, Robert J.; Hayes, Tamara L.; Horak, Fay B.

    2014-01-01

    The Nintendo Wii balance board (WBB) has generated significant interest in its application as a postural control measurement device in both the clinical and (basic, clinical, and rehabilitation) research domains. Although the WBB has been proposed as an alternative to the “gold standard” laboratory-grade force plate, additional research is necessary before the WBB can be considered a valid and reliable center of pressure (CoP) measurement device. In this study, we used the WBB and a laboratory-grade AMTI force plate (AFP) to simultaneously measure the CoP displacement of a controlled dynamic load, which has not been done before. A one-dimensional inverted pendulum was displaced at several different displacement angles and load heights to simulate a variety of postural sway amplitudes and frequencies (<1 Hz). Twelve WBBs were tested to address the issue of inter-device variability. There was a significant effect of sway amplitude, frequency, and direction on the WBB's CoP measurement error, with an increase in error as both sway amplitude and frequency increased and a significantly greater error in the mediolateral (ML) (compared to the anteroposterior (AP)) sway direction. There was no difference in error across the 12 WBB's, supporting low inter-device variability. A linear calibration procedure was then implemented to correct the WBB's CoP signals and reduce measurement error. There was a significant effect of calibration on the WBB's CoP signal accuracy, with a significant reduction in CoP measurement error (quantified by root-mean-squared error) from 2–6 mm (before calibration) to 0.5–2 mm (after calibration). WBB-based CoP signal calibration also significantly reduced the percent error in derived (time-domain) CoP sway measures, from −10.5% (before calibration) to −0.05% (after calibration) (percent errors averaged across all sway measures and in both sway directions). In this study, we characterized the WBB's CoP measurement error under controlled, dynamic conditions and implemented a linear calibration procedure for WBB CoP signals that is recommended to reduce CoP measurement error and provide more reliable estimates of time-domain CoP measures. Despite our promising results, additional work is necessary to understand how our findings translate to the clinical and rehabilitation research domains. Once the WBB's CoP measurement error is fully characterized in human postural sway (which differs from our simulated postural sway in both amplitude and frequency content), it may be used to measure CoP displacement in situations where lower accuracy and precision is acceptable. PMID:25268919

  18. Transcript analysis of the extended hyp-operon in the cyanobacteria Nostoc sp. strain PCC 7120 and Nostoc punctiforme ATCC 29133

    PubMed Central

    2011-01-01

    Background Cyanobacteria harbor two [NiFe]-type hydrogenases consisting of a large and a small subunit, the Hup- and Hox-hydrogenase, respectively. Insertion of ligands and correct folding of nickel-iron hydrogenases require assistance of accessory maturation proteins (encoded by the hyp-genes). The intergenic region between the structural genes encoding the uptake hydrogenase (hupSL) and the accessory maturation proteins (hyp genes) in the cyanobacteria Nostoc PCC 7120 and N. punctiforme were analysed using molecular methods. Findings The five ORFs, located in between the uptake hydrogenase structural genes and the hyp-genes, can form a transcript with the hyp-genes. An identical genomic localization of these ORFs are found in other filamentous, N2-fixing cyanobacterial strains. In N. punctiforme and Nostoc PCC 7120 the ORFs upstream of the hyp-genes showed similar transcript level profiles as hupS (hydrogenase structural gene), nifD (nitrogenase structural gene), hypC and hypF (accessory hydrogenase maturation genes) after nitrogen depletion. In silico analyzes showed that these ORFs in N. punctiforme harbor the same conserved regions as their homologues in Nostoc PCC 7120 and that they, like their homologues in Nostoc PCC 7120, can be transcribed together with the hyp-genes forming a larger extended hyp-operon. DNA binding studies showed interactions of the transcriptional regulators CalA and CalB to the promoter regions of the extended hyp-operon in N. punctiforme and Nostoc PCC 7120. Conclusions The five ORFs upstream of the hyp-genes in several filamentous N2-fixing cyanobacteria have an identical genomic localization, in between the genes encoding the uptake hydrogenase and the maturation protein genes. In N. punctiforme and Nostoc PCC 7120 they are transcribed as one operon and may form transcripts together with the hyp-genes. The expression pattern of the five ORFs within the extended hyp-operon in both Nostoc punctiforme and Nostoc PCC 7120 is similar to the expression patterns of hupS, nifD, hypF and hypC. CalA, a known transcription factor, interacts with the promoter region between hupSL and the five ORFs in the extended hyp-operon in both Nostoc strains. PMID:21672234

  19. Maximization of transcription of the serC (pdxF)-aroA multifunctional operon by antagonistic effects of the cyclic AMP (cAMP) receptor protein-cAMP complex and Lrp global regulators of Escherichia coli K-12.

    PubMed

    Man, T K; Pease, A J; Winkler, M E

    1997-06-01

    The arrangement of the Escherichia coli serC (pdxF) and aroA genes into a cotranscribed multifunctional operon allows coregulation of two enzymes required for the biosynthesis of L-serine, pyridoxal 5'-phosphate, chorismate, and the aromatic amino acids and vitamins. RNase T2 protection assays revealed two major transcripts that were initiated from a promoter upstream from serC (pdxF). Between 80 to 90% of serC (pdxF) transcripts were present in single-gene mRNA molecules that likely arose by Rho-independent termination between serC (pdxF) and aroA. serC (pdxF)-aroA cotranscripts terminated at another Rho-independent terminator near the end of aroA. We studied operon regulation by determining differential rates of beta-galactosidase synthesis in a merodiploid strain carrying a single-copy lambda[phi(serC [pdxF]'-lacZYA)] operon fusion. serC (pdxF) transcription was greatest in bacteria growing in minimal salts-glucose medium (MMGlu) and was reduced in minimal salts-glycerol medium, enriched MMGlu, and LB medium. serC (pdxF) transcription was increased in cya or crp mutants compared to their cya+ crp+ parent in MMGlu or LB medium. In contrast, serC (pdxF) transcription decreased in an lrp mutant compared to its lrp+ parent in MMGlu. Conclusions obtained by using the operon fusion were corroborated by quantitative Western immunoblotting of SerC (PdxF), which was present at around 1,800 dimers per cell in bacteria growing in MMGlu. RNase T2 protection assays of serC (pdxF)-terminated and serC (pdxF)-aroA cotranscript amounts supported the conclusion that the operon was regulated at the transcription level under the conditions tested. Results with a series of deletions upstream of the P(serC (pdxF)) promoter revealed that activation by Lrp was likely direct, whereas repression by the cyclic AMP (cAMP) receptor protein-cAMP complex (CRP-cAMP) was likely indirect, possibly via a repressor whose amount or activity was stimulated by CRP-cAMP.

  20. 20 CFR 10.206 - May an employee who uses leave after an injury later decide to use COP instead?

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... injury later decide to use COP instead? 10.206 Section 10.206 Employees' Benefits OFFICE OF WORKERS... THE FEDERAL EMPLOYEES' COMPENSATION ACT, AS AMENDED Continuation of Pay Eligibility for Cop § 10.206 May an employee who uses leave after an injury later decide to use COP instead? On Form CA-1, an...

  1. 20 CFR 10.206 - May an employee who uses leave after an injury later decide to use COP instead?

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... injury later decide to use COP instead? 10.206 Section 10.206 Employees' Benefits OFFICE OF WORKERS... THE FEDERAL EMPLOYEES' COMPENSATION ACT, AS AMENDED Continuation of Pay Eligibility for Cop § 10.206 May an employee who uses leave after an injury later decide to use COP instead? On Form CA-1, an...

  2. 30 CFR 585.632 - What documents must I submit before I may construct and install facilities under my approved COP?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... construct and install facilities under my approved COP? 585.632 Section 585.632 Mineral Resources BUREAU OF... Approved Cop § 585.632 What documents must I submit before I may construct and install facilities under my approved COP? (a) You must submit to BOEM the documents listed in the following table: Document...

  3. 30 CFR 585.632 - What documents must I submit before I may construct and install facilities under my approved COP?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... construct and install facilities under my approved COP? 585.632 Section 585.632 Mineral Resources BUREAU OF... Approved Cop § 585.632 What documents must I submit before I may construct and install facilities under my approved COP? (a) You must submit to BOEM the documents listed in the following table: Document...

  4. 30 CFR 585.632 - What documents must I submit before I may construct and install facilities under my approved COP?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... construct and install facilities under my approved COP? 585.632 Section 585.632 Mineral Resources BUREAU OF... Approved Cop § 585.632 What documents must I submit before I may construct and install facilities under my approved COP? (a) You must submit to BOEM the documents listed in the following table: Document...

  5. 20 CFR 10.206 - May an employee who uses leave after an injury later decide to use COP instead?

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... injury later decide to use COP instead? 10.206 Section 10.206 Employees' Benefits OFFICE OF WORKERS... THE FEDERAL EMPLOYEES' COMPENSATION ACT, AS AMENDED Continuation of Pay Eligibility for Cop § 10.206 May an employee who uses leave after an injury later decide to use COP instead? On Form CA-1, an...

  6. 50 CFR 23.86 - How can I obtain information on a CoP?

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 50 Wildlife and Fisheries 9 2014-10-01 2014-10-01 false How can I obtain information on a CoP? 23... FAUNA AND FLORA (CITES) CITES Administration § 23.86 How can I obtain information on a CoP? As we receive information on an upcoming CoP from the CITES Secretariat, we will notify the public either...

  7. 50 CFR 23.86 - How can I obtain information on a CoP?

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 50 Wildlife and Fisheries 6 2010-10-01 2010-10-01 false How can I obtain information on a CoP? 23... FAUNA AND FLORA (CITES) CITES Administration § 23.86 How can I obtain information on a CoP? As we receive information on an upcoming CoP from the CITES Secretariat, we will notify the public either...

  8. 50 CFR 23.86 - How can I obtain information on a CoP?

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 50 Wildlife and Fisheries 9 2012-10-01 2012-10-01 false How can I obtain information on a CoP? 23... FAUNA AND FLORA (CITES) CITES Administration § 23.86 How can I obtain information on a CoP? As we receive information on an upcoming CoP from the CITES Secretariat, we will notify the public either...

  9. 50 CFR 23.86 - How can I obtain information on a CoP?

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 50 Wildlife and Fisheries 9 2013-10-01 2013-10-01 false How can I obtain information on a CoP? 23... FAUNA AND FLORA (CITES) CITES Administration § 23.86 How can I obtain information on a CoP? As we receive information on an upcoming CoP from the CITES Secretariat, we will notify the public either...

  10. 50 CFR 23.86 - How can I obtain information on a CoP?

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 50 Wildlife and Fisheries 8 2011-10-01 2011-10-01 false How can I obtain information on a CoP? 23... FAUNA AND FLORA (CITES) CITES Administration § 23.86 How can I obtain information on a CoP? As we receive information on an upcoming CoP from the CITES Secretariat, we will notify the public either...

  11. Concept for a common operational picture in a guidance vehicle

    NASA Astrophysics Data System (ADS)

    Wagner, Boris; Eck, Ralf; Maier, Sebastian

    2017-05-01

    A Common Operational Picture (COP) shows many operational aspects in coded form inside a geodata representation like a map. For building this picture, many specialized groups produce information. Beside the operating forces these are intelligences, logistics, or the own leaders planning group. Operations in which a COP is used typically are disaster management or military actions. An existing software for Interactive Visualization of Integrated Geodata runs on Tablet-PCs, PCs, Digital Map Tables and video walls. It is already used by the Deutsche Führungsakademie (military academy) for the education of staff officers. German civil disaster management agency decided to use the Digital Map Table for their intelligence analysis. In a mobile scenario, however, novel requirements have to be taken into account to adapt the software to the new environment. This paper investigates these requirements as well as the possible adaptions to provide a COP across multiple players on the go. When acting together, the groups do this in a widespread manner. They are physically spread and they use a variety of software and hardware to produce their contribution. This requires hardware to be ruggedized, mobile, and to support a variety of interfaces. The limited bandwidth in such a setting poses the main challenge for the software, which has to synchronize exchanging a minimum of information. Especially for mobile participants, a solution is planned that scales the amount of data (maps/intelligence data) to the available equipment, the upcoming mission, and the underlying theatre. Special focus is laid on a guidance vehicle leading a convoy.

  12. Identification and characterization of a novel outer membrane protein receptor required for hemin utilization in Vibrio vulnificus

    PubMed Central

    Datta, Shreya

    2011-01-01

    Vibrio vulnificus, the cause of septicemia and serious wound infection in humans and fishes, require iron for its pathogenesis. Hemin uptake through the outer membrane receptor, HupA, is one of its many mechanisms by which it acquires iron. We report here the identification of an additional TonB-dependent hemin receptor HvtA, that is needed in conjunction with the HupA protein for optimal hemin utilization. The HvtA protein is significantly homologous to other outer membrane hemin receptors and its expression in trans restored the uptake of hemin and hemoglobin, the latter to a weaker extent, in a mutant strain that was defective in both receptors. Quantitative RT-PCR suggested that transcription of the hvtA gene was iron regulated. The operon containing the hvtA gene is homologous to the operon in V. cholerae containing the hemin receptor gene hutR suggesting a vertical transmission of the hvtA cluster from V. cholerae to V. vulnificus. PMID:22015545

  13. Variability of rRNA Operon Copy Number and Growth Rate Dynamics of Bacillus Isolated from an Extremely Oligotrophic Aquatic Ecosystem

    PubMed Central

    Valdivia-Anistro, Jorge A.; Eguiarte-Fruns, Luis E.; Delgado-Sapién, Gabriela; Márquez-Zacarías, Pedro; Gasca-Pineda, Jaime; Learned, Jennifer; Elser, James J.; Olmedo-Alvarez, Gabriela; Souza, Valeria

    2016-01-01

    The ribosomal RNA (rrn) operon is a key suite of genes related to the production of protein synthesis machinery and thus to bacterial growth physiology. Experimental evidence has suggested an intrinsic relationship between the number of copies of this operon and environmental resource availability, especially the availability of phosphorus (P), because bacteria that live in oligotrophic ecosystems usually have few rrn operons and a slow growth rate. The Cuatro Ciénegas Basin (CCB) is a complex aquatic ecosystem that contains an unusually high microbial diversity that is able to persist under highly oligotrophic conditions. These environmental conditions impose a variety of strong selective pressures that shape the genome dynamics of their inhabitants. The genus Bacillus is one of the most abundant cultivable bacterial groups in the CCB and usually possesses a relatively large number of rrn operon copies (6–15 copies). The main goal of this study was to analyze the variation in the number of rrn operon copies of Bacillus in the CCB and to assess their growth-related properties as well as their stoichiometric balance (N and P content). We defined 18 phylogenetic groups within the Bacilli clade and documented a range of from six to 14 copies of the rrn operon. The growth dynamic of these Bacilli was heterogeneous and did not show a direct relation to the number of operon copies. Physiologically, our results were not consistent with the Growth Rate Hypothesis, since the copies of the rrn operon were decoupled from growth rate. However, we speculate that the diversity of the growth properties of these Bacilli as well as the low P content of their cells in an ample range of rrn copy number is an adaptive response to oligotrophy of the CCB and could represent an ecological mechanism that allows these taxa to coexist. These findings increase the knowledge of the variability in the number of copies of the rrn operon in the genus Bacillus and give insights about the physiology of this bacterial group under extreme oligotrophic conditions. PMID:26779143

  14. Development of High Capacity Split Stirling Cryocooler for HTS

    NASA Astrophysics Data System (ADS)

    Yumoto, Kenta; Nakano, Kyosuke; Hiratsuka, Yoshikatsu

    Sumitomo Heavy Industries, Ltd. (SHI) developed a high-power Stirling-type pulse tube cryocooler for cooling high-temperature superconductor (HTS) devices, such as superconductor motors, superconducting magnetic energy storage (SMES), and fault current limiters. The experimental results of a prototype pulse tube cryocooler were reported in September 2013. For a U-type expander, the cooling capacity was 151 W at 70 K with a compressor input power of 4 kW. Correspondingly, the coefficient of performance (COP) was about 0.038. However, the efficiency of the cryocooler is required to be COP > 0.1 and it was found that, theoretically, it is difficult to further improve the efficiency of a pulse tube cryocooler because the workflow generated at the hot end of the pulse tube cannot be recovered. Therefore, it was decided to change the expander to a free-piston type from a pulse tube type. A prototype was developed and preliminary experiments were conducted. A cooling capacity of 120 W at 70 K with a compressor input power of 2.15 kW with corresponding COP of 0.056, was obtained. The detailed results are reported in this paper.

  15. The combination of platelet count and neutrophil lymphocyte ratio is a predictive factor in patients with esophageal squamous cell carcinoma.

    PubMed

    Feng, Ji-Feng; Huang, Ying; Chen, Qi-Xun

    2014-10-01

    The prognostic value of inflammation indexes in esophageal cancer was not established. In this study, therefore, both prognostic values of Glasgow prognostic score (GPS) and combination of platelet count and neutrophil lymphocyte ratio (COP-NLR) in patients with esophageal squamous cell carcinoma (ESCC) were investigated and compared. This retrospective study included 375 patients who underwent esophagectomy for ESCC. The cancer-specific survival (CSS) was calculated by the Kaplan-Meier method, and the difference was assessed by the log-rank test. The GPS was calculated as follows: patients with elevated C-reactive protein (> 10 mg/l) and hypoalbuminemia (< 35 g/l) were assigned to GPS2. Patients with one or no abnormal value were assigned to GPS1 or GPS0, respectively. The COP-NLR was calculated as follows: patients with elevated platelet count (> 300 × 10(9)/l) and neutrophil lymphocyte ratio (> 3) were assigned to COP-NLR2. Patients with one or no abnormal value were assigned to COP-NLR1 or COP-NLR0, respectively. The 5-year CSS in patients with GPS0, 1, and 2 was 50.0%, 27.0%, and 12.5%, respectively (P < .001). The 5-year CSS in patients with COP-NLR0, 1, and 2 was 51.8%, 27.0%, and 11.6%, respectively (P < .001). Multivariate analysis showed that both GPS (P = .003) and COP-NLR (P = .003) were significant predictors in such patients. In addition, our study demonstrated a similar hazard ratio (HR) between COP-NLR and GPS (HR = 1.394 vs HR = 1.367). COP-NLR is an independent predictive factor in patients with ESCC. We conclude that COP-NLR predicts survival in ESCC similar to GPS.

  16. Diametral tensile strength and film thickness of an experimental dental luting agent derived from castor oil.

    PubMed

    Carmello, Juliana Cabrini; Fais, Laiza Maria Grassi; Ribeiro, Lígia Nunes de Moraes; Claro Neto, Salvador; Guaglianoni, Dalton Geraldo; Pinelli, Lígia Antunes Pereira

    2012-02-01

    The need to develop new dental luting agents in order to improve the success of treatments has greatly motivated research. The aim of this study was to evaluate the diametral tensile strength (DTS) and film thickness (FT) of an experimental dental luting agent derived from castor oil (COP) with or without addition of different quantities of filler (calcium carbonate - CaCO3). Eighty specimens were manufactured (DTS N=40; FT N=40) and divided into 4 groups: Pure COP; COP 10%; COP 50% and zinc phosphate (control). The cements were mixed according to the manufacturers' recommendations and submitted to the tests. The DTS test was performed in the MTS 810 testing machine (10 KN, 0.5 mm/min). For FT test, the cements were sandwiched between two glass plates (2 cm²) and a load of 15 kg was applied vertically on the top of the specimen for 10 min. The data were analyzed by means of one-way ANOVA and Tukey's test (α=0.05). The values of DTS (MPa) were: Pure COP- 10.94 ± 1.30; COP 10%- 30.06 ± 0.64; COP 50%- 29.87 ± 0.27; zinc phosphate- 4.88 ± 0.96. The values of FT (µm) were: Pure COP- 31.09 ± 3.16; COP 10%- 17.05 ± 4.83; COP 50%- 13.03 ± 4.83; Zinc Phosphate- 20.00 ± 0.12. One-way ANOVA showed statistically significant differences among the groups (DTS - p=1.01E-40; FT - p=2.4E-10). The experimental dental luting agent with 50% of filler showed the best diametral tensile strength and film thickness.

  17. Diametral tensile strength and film thickness of an experimental dental luting agent derived from castor oil

    PubMed Central

    CARMELLO, Juliana Cabrini; FAIS, Laiza Maria Grassi; RIBEIRO, Lígia Nunes de Moraes; CLARO NETO, Salvador; GUAGLIANONI, Dalton Geraldo; PINELLI, Lígia Antunes Pereira

    2012-01-01

    The need to develop new dental luting agents in order to improve the success of treatments has greatly motivated research. Objective The aim of this study was to evaluate the diametral tensile strength (DTS) and film thickness (FT) of an experimental dental luting agent derived from castor oil (COP) with or without addition of different quantities of filler (calcium carbonate - CaCO3). Material and Methods Eighty specimens were manufactured (DTS N=40; FT N=40) and divided into 4 groups: Pure COP; COP 10%; COP 50% and zinc phosphate (control). The cements were mixed according to the manufacturers' recommendations and submitted to the tests. The DTS test was performed in the MTS 810 testing machine (10 KN, 0.5 mm/min). For FT test, the cements were sandwiched between two glass plates (2 cm2) and a load of 15 kg was applied vertically on the top of the specimen for 10 min. The data were analyzed by means of one-way ANOVA and Tukey's test (α=0.05). Results The values of DTS (MPa) were: Pure COP- 10.94±1.30; COP 10%- 30.06±0.64; COP 50%- 29.87±0.27; zinc phosphate- 4.88±0.96. The values of FT (µm) were: Pure COP- 31.09±3.16; COP 10%- 17.05±4.83; COP 50%- 13.03±4.83; Zinc Phosphate- 20.00±0.12. One-way ANOVA showed statistically significant differences among the groups (DTS - p=1.01E-40; FT - p=2.4E-10). Conclusion The experimental dental luting agent with 50% of filler showed the best diametral tensile strength and film thickness. PMID:22437672

  18. Reproducibility and diagnostic performance of shear wave elastography in evaluating breast solid mass.

    PubMed

    Hong, Sun; Woo, Ok Hee; Shin, Hye Seon; Hwang, Soon-Young; Cho, Kyu Ran; Seo, Bo Kyoung

    Shear wave elastography (SWE) was performed independently by two radiologists in 264 solid breast masses. The images were reviewed for color overlay pattern (COP) classification by the two radiologists, double blinded to any information. The interobserver agreement of the COP was almost perfect (κ=0.908) and high in E max (ICC=0.89). The AUC value of the COP (0.954) was significantly higher than that of E max (0.915) (p=0.002) but not significantly different from that of E max combined with COP (0.957) (p=0.098). The SWE color overlay pattern and E max of breast masses were highly reproducible. The COP had better diagnostic ability than E max , suggesting that COP may be a more reliable parameter for solid breast mass evaluation. Copyright © 2017 Elsevier Inc. All rights reserved.

  19. What's in a name? Tensions between formal and informal communities of practice among regional subspecialty cancer surgeons.

    PubMed

    Kitto, Simon C; Grant, Rachel E; Peller, Jennifer; Moulton, Carol-Anne; Gallinger, Steven

    2018-03-01

    In 2007 the Cancer Care Ontario Hepatobiliary-Pancreatic (HPB) Community of Practice was formed during the wake of provincial regionalization of HPB services in Ontario, Canada. Despite being conceptualized within the literature as an educational intervention, communities of practice (CoP) are increasingly being adopted in healthcare as quality improvement initiatives. A qualitative case study approach using in-depth interviews and document analysis was employed to gain insight into the perceptions and attitudes of the HPB surgeons in the CoP. This study demonstrates how an engineered formal or idealized structure of a CoP was created in tension with the natural CoPs that HPB surgeons identified with during and after their training. This tension contributed to the inactive and/or marginal participation by some of the surgeons in the CoP. The findings of this study represent a cautionary tale for such future engineering attempts in two distinct ways: (1) a CoP in surgery cannot simply be created by regulatory agencies, rather they need to be supported in a way to evolve naturally, and (2) when the concept of CoPs is co-opted by governing bodies, it does not necessarily capture the power and potential of situated learning. To ensure CoP sustainability and effectiveness, we suggest that both core and peripheral members need to be more directly involved at the inception of the COP in terms of design, organization, implementation and ongoing management.

  20. A quantitative study of the benefits of co-regulation using the spoIIA operon as an example

    PubMed Central

    Iber, Dagmar

    2006-01-01

    The distribution of most genes is not random, and functionally linked genes are often found in clusters. Several theories have been put forward to explain the emergence and persistence of operons in bacteria. Careful analysis of genomic data favours the co-regulation model, where gene organization into operons is driven by the benefits of coordinated gene expression and regulation. Direct evidence that coexpression increases the individual's fitness enough to ensure operon formation and maintenance is, however, still lacking. Here, a previously described quantitative model of the network that controls the transcription factor σF during sporulation in Bacillus subtilis is employed to quantify the benefits arising from both organization of the sporulation genes into the spoIIA operon and from translational coupling. The analysis shows that operon organization, together with translational coupling, is important because of the inherent stochastic nature of gene expression, which skews the ratios between protein concentrations in the absence of co-regulation. The predicted impact of different forms of gene regulation on fitness and survival agrees quantitatively with published sporulation efficiencies. PMID:16924264

  1. A quantitative study of the benefits of co-regulation using the spoIIA operon as an example.

    PubMed

    Iber, Dagmar

    2006-01-01

    The distribution of most genes is not random, and functionally linked genes are often found in clusters. Several theories have been put forward to explain the emergence and persistence of operons in bacteria. Careful analysis of genomic data favours the co-regulation model, where gene organization into operons is driven by the benefits of coordinated gene expression and regulation. Direct evidence that coexpression increases the individual's fitness enough to ensure operon formation and maintenance is, however, still lacking. Here, a previously described quantitative model of the network that controls the transcription factor sigma(F) during sporulation in Bacillus subtilis is employed to quantify the benefits arising from both organization of the sporulation genes into the spoIIA operon and from translational coupling. The analysis shows that operon organization, together with translational coupling, is important because of the inherent stochastic nature of gene expression, which skews the ratios between protein concentrations in the absence of co-regulation. The predicted impact of different forms of gene regulation on fitness and survival agrees quantitatively with published sporulation efficiencies.

  2. 20 CFR 10.217 - Is COP charged if the employee continues to work, but in a different job that pays less?

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 20 Employees' Benefits 1 2012-04-01 2012-04-01 false Is COP charged if the employee continues to... COMPENSATION UNDER THE FEDERAL EMPLOYEES' COMPENSATION ACT, AS AMENDED Continuation of Pay Calculation of Cop § 10.217 Is COP charged if the employee continues to work, but in a different job that pays less? If...

  3. 20 CFR 10.217 - Is COP charged if the employee continues to work, but in a different job that pays less?

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 20 Employees' Benefits 1 2014-04-01 2012-04-01 true Is COP charged if the employee continues to... COMPENSATION UNDER THE FEDERAL EMPLOYEES' COMPENSATION ACT, AS AMENDED Continuation of Pay Calculation of Cop § 10.217 Is COP charged if the employee continues to work, but in a different job that pays less? If...

  4. 20 CFR 10.217 - Is COP charged if the employee continues to work, but in a different job that pays less?

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 20 Employees' Benefits 1 2013-04-01 2012-04-01 true Is COP charged if the employee continues to... COMPENSATION UNDER THE FEDERAL EMPLOYEES' COMPENSATION ACT, AS AMENDED Continuation of Pay Calculation of Cop § 10.217 Is COP charged if the employee continues to work, but in a different job that pays less? If...

  5. 20 CFR 10.217 - Is COP charged if the employee continues to work, but in a different job that pays less?

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 20 Employees' Benefits 1 2010-04-01 2010-04-01 false Is COP charged if the employee continues to... COMPENSATION UNDER THE FEDERAL EMPLOYEES' COMPENSATION ACT, AS AMENDED Continuation of Pay Calculation of Cop § 10.217 Is COP charged if the employee continues to work, but in a different job that pays less? If...

  6. 20 CFR 10.217 - Is COP charged if the employee continues to work, but in a different job that pays less?

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 20 Employees' Benefits 1 2011-04-01 2011-04-01 false Is COP charged if the employee continues to... COMPENSATION UNDER THE FEDERAL EMPLOYEES' COMPENSATION ACT, AS AMENDED Continuation of Pay Calculation of Cop § 10.217 Is COP charged if the employee continues to work, but in a different job that pays less? If...

  7. Performance analysis for minimally nonlinear irreversible refrigerators at finite cooling power

    NASA Astrophysics Data System (ADS)

    Long, Rui; Liu, Zhichun; Liu, Wei

    2018-04-01

    The coefficient of performance (COP) for general refrigerators at finite cooling power have been systematically researched through the minimally nonlinear irreversible model, and its lower and upper bounds in different operating regions have been proposed. Under the tight coupling conditions, we have calculated the universal COP bounds under the χ figure of merit in different operating regions. When the refrigerator operates in the region with lower external flux, we obtained the general bounds (0 < ε <(√{ 9 + 8εC } - 3) / 2) under the χ figure of merit. We have also calculated the universal bounds for maximum gain in COP under different operating regions to give a further insight into the COP gain with the cooling power away from the maximum one. When the refrigerator operates in the region located between maximum cooling power and maximum COP with lower external flux, the upper bound for COP and the lower bound for relative gain in COP present large values, compared to a relative small loss from the maximum cooling power. If the cooling power is the main objective, it is desirable to operate the refrigerator at a slightly lower cooling power than at the maximum one, where a small loss in the cooling power induces a much larger COP enhancement.

  8. An assessment of antioxidant status in patients with carbon monoxide poisoning

    PubMed Central

    Zengin, Suat; A, Behcet; Karta, Sahin; Can, Basri; Orkmez, Mustafa; Taskin, Abdullah; Lok, Ugur; Gulen, Bediha; Yildirim, Cuma; Taysi, Seyithan

    2014-01-01

    BACKGROUND: Carbon monoxide poisoning (COP) is an important cause of mortality and morbidity worldwide. This study was to investigate the levels of serum paraoxonase (PON), arylesterase (ARYL), ceruloplasmin (Cp), and sulfhydryl (-SH) in the treatment of COP, and to further understand the pathophysiology of COP. METHODS: This prospective study comprised 107 individuals with COP (group 1) and 50 healthy volunteers (group 2). Serum, plasma, and erythrocyte samples were taken on admission from all participants with COP. This process was repeated in the 90th and 180th minutes of treatment. Samples were taken from the control group only once. The levels of plasma PON, ARYL, Cp activity and -SH were measured in both groups. RESULTS: Age, gender, and carboxyhemoglobin level were not correlated with PON, ARYL, Cp, and -SH levels. PON, ARYL, and -SH levels were significantly decreased in group 1 compared with group 2. Conversely, Cp was significantly elevated in group 1 in contrast to group 2. Although ARYL was lower on admission in patients with COP than that was observed in the 90th and 180th minutes (P<0.001), Cp was higher on admission than at the other time points (P<0.001). CONCLUSIONS: Participants with COP had decreased levels of antioxidants (PON, ARLY, and -SH). COP represses the antioxidant system. PMID:25215155

  9. Inhibition of cholesterol oxidation products (COPs) formation in emulsified porcine patties by phenolic-rich avocado (Persea americana Mill.) extracts.

    PubMed

    Rodríguez-Carpena, Javier-Germán; Morcuende, David; Petrón, María Jesus; Estévez, Mario

    2012-03-07

    The effect of phenolic-rich extracts from avocado peel on the formation of cholesterol oxidation products (COPs) in porcine patties subjected to cooking and chill storage was studied. Eight COPs (7α-hydroxycholesterol, 7β-hydroxycholesterol, 7-ketocholesterol, 20α-hydroxycholesterol, 25-hydroxycholesterol, cholestanetriol, 5,6β-epoxycholesterol, and 5,6α-epoxycholesterol) were identified and quantified by GC-MS. The addition of avocado extracts (∼600 GAE/kg patty) to patties significantly inhibited the formation of COPs during cooking. Cooked control (C) patties contained a larger variety and greater amounts of COPs than the avocado-treated (T) counterparts. COPs sharply increased in cooked patties during the subsequent chilled storage. This increase was significantly higher in C patties than in the T patties. Interestingly, the amount of COPs in cooked and chilled T patties was similar to those found in cooked C patties. The mechanisms implicated in cholesterol oxidation in a processed meat product, the protective effect of avocado phenolics, and the potential implication of lipid and protein oxidation are thoroughly described in the present paper.

  10. Phenotypic Characterization of a copA Mutant of Neisseria gonorrhoeae Identifies a Link between Copper and Nitrosative Stress

    PubMed Central

    Djoko, Karrera Y.; Franiek, Jessica A.; Edwards, Jennifer L.; Falsetta, Megan L.; Kidd, Stephen P.; Potter, Adam J.; Chen, Nathan H.; Apicella, Michael A.; Jennings, Michael P.

    2012-01-01

    NGO0579 is annotated copA in the Neisseria gonorrhoeae chromosome, suggesting that it encodes a cation-transporting ATPase specific for copper ions. Compared to wild-type cells, a copA mutant was more sensitive to killing by copper ions but not to other transition metals. The mutant also accumulated a greater amount of copper, consistent with the predicted role of CopA as a copper efflux pump. The copA mutant showed a reduced ability to invade and survive within human cervical epithelial cells, although its ability to form a biofilm on the surface of these cells was not significantly different from that of the wild type. In the presence of copper, the copA mutant exhibited increased sensitivity to killing by nitrite or nitric oxide. Therefore, we concluded that copper ion efflux catalyzed by CopA is linked to the nitrosative stress defense system of Neisseria gonorrhoeae. These observations suggest that copper may exert its effects as an antibacterial agent in the innate immune system via an interaction with reactive nitrogen species. PMID:22184419

  11. Rules for the recognition of dilysine retrieval motifs by coatomer

    PubMed Central

    Ma, Wenfu; Goldberg, Jonathan

    2013-01-01

    Cytoplasmic dilysine motifs on transmembrane proteins are captured by coatomer α-COP and β′-COP subunits and packaged into COPI-coated vesicles for Golgi-to-ER retrieval. Numerous ER/Golgi proteins contain K(x)Kxx motifs, but the rules for their recognition are unclear. We present crystal structures of α-COP and β′-COP bound to a series of naturally occurring retrieval motifs—encompassing KKxx, KxKxx and non-canonical RKxx and viral KxHxx sequences. Binding experiments show that α-COP and β′-COP have generally the same specificity for KKxx and KxKxx, but only β′-COP recognizes the RKxx signal. Dilysine motif recognition involves lysine side-chain interactions with two acidic patches. Surprisingly, however, KKxx and KxKxx motifs bind differently, with their lysine residues transposed at the binding patches. We derive rules for retrieval motif recognition from key structural features: the reversed binding modes, the recognition of the C-terminal carboxylate group which enforces lysine positional context, and the tolerance of the acidic patches for non-lysine residues. PMID:23481256

  12. An extracytoplasmic function sigma factor-dependent periplasmic glutathione peroxidase is involved in oxidative stress response of Shewanella oneidensis

    DOE PAGES

    Dai, Jingcheng; Wei, Hehong; Tian, Chunyuan; ...

    2015-01-01

    Background: Bacteria use alternative sigma factors (σs) to regulate condition-specific gene expression for survival and Shewanella harbors multiple ECF (extracytoplasmic function) σ genes and cognate anti-sigma factor genes. Here we comparatively analyzed two of the rpoE-like operons in the strain MR-1: rpoE-rseA-rseB-rseC and rpoE2-chrR. Results: RpoE was important for bacterial growth at low and high temperatures, in the minimal medium, and high salinity. The degP/htrA orthologue, required for growth of Escherichia coli and Pseudomonas aeruginosa at high temperature, is absent in Shewanella, while the degQ gene is RpoE-regulated and is required for bacterial growth at high temperature. RpoE2 was essentialmore » for the optimal growth in oxidative stress conditions because the rpoE2 mutant was sensitive to hydrogen peroxide and paraquat. The operon encoding a ferrochelatase paralogue (HemH2) and a periplasmic glutathione peroxidase (PgpD) was identified as RpoE2-dependent. PgpD exhibited higher activities and played a more important role in the oxidative stress responses than the cytoplasmic glutathione peroxidase CgpD under tested conditions. The rpoE2-chrR operon and the identified regulon genes, including pgpD and hemH2, are coincidently absent in several psychrophilic and/or deep-sea Shewanella strains. Conclusion: In S. oneidensis MR-1, the RpoE-dependent degQ gene is required for optimal growth under high temperature. The rpoE2 and RpoE2-dependent pgpD gene encoding a periplasmic glutathione peroxidase are involved in oxidative stress responses. But rpoE2 is not required for bacterial growth at low temperature and it even affected bacterial growth under salt stress, indicating that there is a tradeoff between the salt resistance and RpoE2-mediated oxidative stress responses.« less

  13. An extracytoplasmic function sigma factor-dependent periplasmic glutathione peroxidase is involved in oxidative stress response of Shewanella oneidensis

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

    Dai, Jingcheng; Wei, Hehong; Tian, Chunyuan

    Background: Bacteria use alternative sigma factors (σs) to regulate condition-specific gene expression for survival and Shewanella harbors multiple ECF (extracytoplasmic function) σ genes and cognate anti-sigma factor genes. Here we comparatively analyzed two of the rpoE-like operons in the strain MR-1: rpoE-rseA-rseB-rseC and rpoE2-chrR. Results: RpoE was important for bacterial growth at low and high temperatures, in the minimal medium, and high salinity. The degP/htrA orthologue, required for growth of Escherichia coli and Pseudomonas aeruginosa at high temperature, is absent in Shewanella, while the degQ gene is RpoE-regulated and is required for bacterial growth at high temperature. RpoE2 was essentialmore » for the optimal growth in oxidative stress conditions because the rpoE2 mutant was sensitive to hydrogen peroxide and paraquat. The operon encoding a ferrochelatase paralogue (HemH2) and a periplasmic glutathione peroxidase (PgpD) was identified as RpoE2-dependent. PgpD exhibited higher activities and played a more important role in the oxidative stress responses than the cytoplasmic glutathione peroxidase CgpD under tested conditions. The rpoE2-chrR operon and the identified regulon genes, including pgpD and hemH2, are coincidently absent in several psychrophilic and/or deep-sea Shewanella strains. Conclusion: In S. oneidensis MR-1, the RpoE-dependent degQ gene is required for optimal growth under high temperature. The rpoE2 and RpoE2-dependent pgpD gene encoding a periplasmic glutathione peroxidase are involved in oxidative stress responses. But rpoE2 is not required for bacterial growth at low temperature and it even affected bacterial growth under salt stress, indicating that there is a tradeoff between the salt resistance and RpoE2-mediated oxidative stress responses.« less

  14. Transcriptional Regulation of Aggregatibacter actinomycetemcomitans lsrACDBFG and lsrRK Operons and Their Role in Biofilm Formation

    PubMed Central

    Torres-Escobar, Ascención; Juárez-Rodríguez, María Dolores; Lamont, Richard J.

    2013-01-01

    Autoinducer-2 (AI-2) is required for biofilm formation and virulence of the oral pathogen Aggregatibacter actinomycetemcomitans, and we previously showed that lsrB codes for a receptor for AI-2. The lsrB gene is expressed as part of the lsrACDBFG operon, which is divergently transcribed from an adjacent lsrRK operon. In Escherichia coli, lsrRK encodes a repressor and AI-2 kinase that function to regulate lsrACDBFG. To determine if lsrRK controls lsrACDBFG expression and influences biofilm growth of A. actinomycetemcomitans, we first defined the promoters for each operon. Transcriptional reporter plasmids containing the 255-bp lsrACDBFG-lsrRK intergenic region (IGR) fused to lacZ showed that essential elements of lsrR promoter reside 89 to 255 bp upstream from the lsrR start codon. Two inverted repeat sequences that represent potential binding sites for LsrR and two sequences resembling the consensus cyclic AMP receptor protein (CRP) binding site were identified in this region. Using electrophoretic mobility shift assay (EMSA), purified LsrR and CRP proteins were shown to bind probes containing these sequences. Surprisingly, the 255-bp IGR did not contain the lsrA promoter. Instead, a fragment encompassing nucleotides +1 to +159 of lsrA together with the 255-bp IGR was required to promote lsrA transcription. This suggests that a region within the lsrA coding sequence influences transcription, or alternatively that the start codon of A. actinomycetemcomitans lsrA has been incorrectly annotated. Transformation of ΔlsrR, ΔlsrK, ΔlsrRK, and Δcrp deletion mutants with lacZ reporters containing the lsrA or lsrR promoter showed that LsrR negatively regulates and CRP positively regulates both lsrACDBFG and lsrRK. However, in contrast to what occurs in E. coli, deletion of lsrK had no effect on the transcriptional activity of the lsrA or lsrR promoters, suggesting that another kinase may be capable of phosphorylating AI-2 in A. actinomycetemcomitans. Finally, biofilm formation of the ΔlsrR, ΔlsrRK, and Δcrp mutants was significantly reduced relative to that of the wild type, indicating that proper regulation of the lsr locus is required for optimal biofilm growth by A. actinomycetemcomitans. PMID:23104800

  15. Synergic role of the two ars operons in arsenic tolerance in Pseudomonas putida KT2440.

    PubMed

    Fernández, Matilde; Udaondo, Zulema; Niqui, José-Luis; Duque, Estrella; Ramos, Juan-Luis

    2014-10-01

    The chromosome of Pseudomonas putida KT2440 carries two clusters of genes, denoted ars1 and ars2, that are annotated as putative arsenic resistance operons. In this work, we present evidence that both operons encode functional arsenic-response regulatory genes as well as arsenic extrusion systems that confer resistance to both arsenite [As(III)] and arsenate [As(V)]. Transcriptional fusions of P(ars1) and P(ars2) to lacZ revealed that expression of both operons was induced by arsenite and arsenate. We generated single mutants in ars1 and ars2, which showed lower resistance to arsenic than the wild-type strain. A double ars1/ars2 was found to be highly sensitive to arsenic. Minimum inhibitory concentrations (MICs) for single mutants decreased two- to fourfold with respect to the parental strain, while in the double mutant the MIC decreased 128-fold for arsenite and 32-fold for arsenate. Bioinformatic analysis revealed that the ars2 resistance operon is part of the core genome of P. putida, while the ars1 operon appears to only occur in the KT2440 strain, suggesting that ars1 was acquired by horizontal gene transfer. The presence of ars1 in KT2440 may explain why it exhibits higher resistance to arsenic than other P. putida strains, which bear only the ars2 operon.

  16. OperomeDB: A Database of Condition-Specific Transcription Units in Prokaryotic Genomes.

    PubMed

    Chetal, Kashish; Janga, Sarath Chandra

    2015-01-01

    Background. In prokaryotic organisms, a substantial fraction of adjacent genes are organized into operons-codirectionally organized genes in prokaryotic genomes with the presence of a common promoter and terminator. Although several available operon databases provide information with varying levels of reliability, very few resources provide experimentally supported results. Therefore, we believe that the biological community could benefit from having a new operon prediction database with operons predicted using next-generation RNA-seq datasets. Description. We present operomeDB, a database which provides an ensemble of all the predicted operons for bacterial genomes using available RNA-sequencing datasets across a wide range of experimental conditions. Although several studies have recently confirmed that prokaryotic operon structure is dynamic with significant alterations across environmental and experimental conditions, there are no comprehensive databases for studying such variations across prokaryotic transcriptomes. Currently our database contains nine bacterial organisms and 168 transcriptomes for which we predicted operons. User interface is simple and easy to use, in terms of visualization, downloading, and querying of data. In addition, because of its ability to load custom datasets, users can also compare their datasets with publicly available transcriptomic data of an organism. Conclusion. OperomeDB as a database should not only aid experimental groups working on transcriptome analysis of specific organisms but also enable studies related to computational and comparative operomics.

  17. Uptake of chitosan-derived D-glucosamine oligosaccharides in Streptomyces coelicolor A3(2).

    PubMed

    Viens, Pascal; Dubeau, Marie-Pierre; Kimura, Akane; Desaki, Yoshitake; Shinya, Tomonori; Shibuya, Naoto; Saito, Akihiro; Brzezinski, Ryszard

    2015-05-01

    The csnR gene, localized at the beginning of an operon, csnR-K, which organization is conserved through many actinomycete genomes, was previously shown to repress the transcription of the chitosanase gene csnA in Streptomyces lividans. However, knowledge on the function of the whole csnR-K operon in the metabolism of chitosan (an N-deacetylated derivative of chitin) remained limited. Mutants of S. coelicolor A3(2) harboring partial or total deletions of the csnR-K operon were analyzed for their capacity to uptake glucosamine oligosaccharides (GlcN)n. The csnR-K operon was autoregulated by CsnR repressor and its transcription was inducible by GlcN oligosaccharides. The operon controlled the uptake of GlcN oligosaccharides in S. coelicolor A3(2), with a minor contribution to the consumption of monomeric GlcN but not chitin-related N-acetylated derivatives. The deletion of the whole operon abolished the uptake of GlcN oligosaccharides. The CsnEFG transporter encoded by this operon is the front door for the assimilation of chitosan-derived hydrolysis products in S. coelicolor A3(2). The ATP-binding component MsiK was essential for CsnEFG transport function. Also, deletion of msiK abolished the induction of csnA transcription by GlcN oligosaccharides. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  18. An operon for production of bioactive gibberellin A4 phytohormone with wide distribution in the bacterial rice leaf streak pathogen Xanthomonas oryzae pv. oryzicola.

    PubMed

    Nagel, Raimund; Turrini, Paula C G; Nett, Ryan S; Leach, Jan E; Verdier, Valérie; Van Sluys, Marie-Anne; Peters, Reuben J

    2017-05-01

    Phytopathogens have developed elaborate mechanisms to attenuate the defense response of their host plants, including convergent evolution of complex pathways for production of the GA phytohormones, which were actually first isolated from the rice fungal pathogen Gibberella fujikuroi. The rice bacterial pathogen Xanthomonas oryzae pv. oryzicola (Xoc) has been demonstrated to contain a biosynthetic operon with cyclases capable of producing the universal GA precursor ent-kaurene. Genetic (knock-out) studies indicate that the derived diterpenoid serves as a virulence factor for this rice leaf streak pathogen, serving to reduce the jasmonic acid-mediated defense response. Here the functions of the remaining genes in the Xoc operon are elucidated and the distribution of the operon in X. oryzae is investigated in over 100 isolates. The Xoc operon leads to production of the bioactive GA 4 , an additional step beyond production of the penultimate precursor GA 9 mediated by the homologous operons recently characterized from rhizobia. Moreover, this GA biosynthetic operon was found to be widespread in Xoc (> 90%), but absent in the other major X. oryzae pathovar. These results indicate selective pressure for production of GA 4 in the distinct lifestyle of Xoc, and the importance of GA to both fungal and bacterial pathogens of rice. © 2017 The Authors. New Phytologist © 2017 New Phytologist Trust.

  19. An operon for production of bioactive gibberellin A4 phytohormone with wide distribution in the bacterial rice leaf streak pathogen Xanthomonas oryzae pv. oryzicola

    PubMed Central

    Nagel, Raimund; Turrini, Paula C. G.; Nett, Ryan S.; Leach, Jan E.; Verdier, Valérie; Van Sluys, Marie-Anne; Peters, Reuben J.

    2016-01-01

    Summary Phytopathogens have developed elaborate mechanisms to attenuate the defense response of their host plants, including convergent evolution of complex pathways for production of the gibberellin (GA) phytohormones, which were actually first isolated from the rice fungal pathogen Gibberella fujikuroi. The rice bacterial pathogen Xanthomonas oryzae pv. oryzicola (Xoc) has been demonstrated to contain a biosynthetic operon with cyclases capable of producing the universal GA precursor ent-kaurene. Genetic (knock-out) studies indicate that the derived diterpenoid serves as a virulence factor for this rice leaf streak pathogen, serving to reduce the jasmonic acid (JA) mediated defense response.Here the function of the remaining genes in the Xoc operon are elucidated and the distribution of the operon in X. oryzae investigated in over 100 isolates.The Xoc operon leads to production of the bioactive GA4, an additional step beyond production of the penultimate precursor GA9 mediated by the homologous operons recently characterized from rhizobia. Moreover, this GA biosynthetic operon was found to be widespread in Xoc (>90%), but absent in the other major oryzae pathovar.These results indicate selective pressure for production of GA4 in the distinct lifestyle of Xoc, and the importance of GA to both fungal and bacterial pathogens of rice. PMID:28134995

  20. Navy Network Dependability: Models, Metrics, and Tools

    DTIC Science & Technology

    2010-01-01

    different COP servers. The COP Synchronization Tool (CST) is the preferred method of exchanging data between COP servers: A critical component of COP...ASW mission’s equipment strings. A major difference in results between the new model and the old one is that the new one is far less optimistic about...understand why perceptions about the dependability (e.g., availability) of networks from users’ (e.g., sailors) per- spectives sometimes differ from the

  1. Understanding balance differences in individuals with multiple sclerosis with mild disability: an investigation of differences in sensory feedback on postural control during a Romberg task.

    PubMed

    Denommé, Luke T; Mandalfino, Patricia; Cinelli, Michael E

    2014-06-01

    A major presenting symptom in 'individuals with multiple sclerosis with mild balance disability' (IwMS) is poor postural control, resulting from slowed spinal somatosensory conduction. Postural control deficits in IwMS are most apparent when vision is removed and the base of support is reduced such is the case during tandem and single support stances. The current study used center of pressure (COP) measurements to determine whether postural control differences exist between IwMS and either 'healthy age-matched individuals' (HAMI) or 'community-dwelling older adults' (OA). Postural control was evaluated using a Romberg standing task, which required participants to stand with their feet together and hands by their sides for 45 s with either their eyes open or closed. Results revealed that COP velocity root mean square was greater in IwMS and their COP position was closer to their self-selected maximum stability limits (e.g., greater Standing Index proportion) when vision was removed compared to HAMI. Conversely, IwMS displayed similar postural control characteristics to OA. The current study highlights two novel findings: (1) the utility of novel COP measurements to assess differences in the level of postural control in IwMS; and (2) the benefit of assessing postural control levels in IwMS to not only a population with a fully intact and functional postural control system (HAMI) but also to another population that is thought to experience postural control deficits (OA).

  2. Quantifying center of pressure variability in chondrodystrophoid dogs.

    PubMed

    Blau, S R; Davis, L M; Gorney, A M; Dohse, C S; Williams, K D; Lim, J-H; Pfitzner, W G; Laber, E; Sawicki, G S; Olby, N J

    2017-08-01

    The center of pressure (COP) position reflects a combination of proprioceptive, motor and mechanical function. As such, it can be used to quantify and characterize neurologic dysfunction. The aim of this study was to describe and quantify the movement of COP and its variability in healthy chondrodystrophoid dogs while walking to provide a baseline for comparison to dogs with spinal cord injury due to acute intervertebral disc herniations. Fifteen healthy adult chondrodystrophoid dogs were walked on an instrumented treadmill that recorded the location of each dog's COP as it walked. Center of pressure (COP) was referenced from an anatomical marker on the dogs' back. The root mean squared (RMS) values of changes in COP location in the sagittal (y) and horizontal (x) directions were calculated to determine the range of COP variability. Three dogs would not walk on the treadmill. One dog was too small to collect interpretable data. From the remaining 11 dogs, 206 trials were analyzed. Mean RMS for change in COPx per trial was 0.0138 (standard deviation, SD 0.0047) and for COPy was 0.0185 (SD 0.0071). Walking speed but not limb length had a significant effect on COP RMS. Repeat measurements in six dogs had high test retest consistency in the x and fair consistency in the y direction. In conclusion, COP variability can be measured consistently in dogs, and a range of COP variability for normal chondrodystrophoid dogs has been determined to provide a baseline for future studies on dogs with spinal cord injury. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. A Numbers Game: Ribosome Densities, Bacterial Growth, and Antibiotic-Mediated Stasis and Death

    PubMed Central

    McCall, Ingrid C.; Perrot, Véronique; Weiss, Howard; Ovesepian, Armen; Baquero, Fernando

    2017-01-01

    ABSTRACT We postulate that the inhibition of growth and low rates of mortality of bacteria exposed to ribosome-binding antibiotics deemed bacteriostatic can be attributed almost uniquely to these drugs reducing the number of ribosomes contributing to protein synthesis, i.e., the number of effective ribosomes. We tested this hypothesis with Escherichia coli K-12 MG1655 and constructs that had been deleted for 1 to 6 of the 7 rRNA (rrn) operons. In the absence of antibiotics, constructs with fewer rrn operons have lower maximum growth rates and longer lag phases than those with more ribosomal operons. In the presence of the ribosome-binding “bacteriostatic” antibiotics tetracycline, chloramphenicol, and azithromycin, E. coli strains with 1 and 2 rrn operons are killed at a substantially higher rate than those with more rrn operons. This increase in the susceptibility of E. coli with fewer rrn operons to killing by ribosome-targeting bacteriostatic antibiotics is not reflected in their greater sensitivity to killing by the bactericidal antibiotic ciprofloxacin, which does not target ribosomes, but also to killing by gentamicin, which does. Finally, when such strains are exposed to these ribosome-targeting bacteriostatic antibiotics, the time before these bacteria start to grow again when the drugs are removed, referred to as the post-antibiotic effect (PAE), is markedly greater for constructs with fewer rrn operons than for those with more rrn operons. We interpret the results of these other experiments reported here as support for the hypothesis that the reduction in the effective number of ribosomes due to binding to these structures provides a sufficient explanation for the action of bacteriostatic antibiotics that target these structures. PMID:28174311

  4. Identification and characterization of transcripts from the biotin biosynthetic operon of Bacillus subtilis.

    PubMed Central

    Perkins, J B; Bower, S; Howitt, C L; Yocum, R R; Pero, J

    1996-01-01

    Northern (RNA) blot analysis of the Bacillus subtilis biotin operon, bioWAFDBIorf2, detected at least two steady-state polycistronic transcripts initiated from a putative vegetative (Pbio) promoter that precedes the operon, i.e., a full-length 7.2-kb transcript covering the entire operon and a more abundant 5.1-kb transcript covering just the first five genes of the operon. Biotin and the B. subtilis birA gene product regulated synthesis of the transcripts. Moreover, replacing the putative Pbio promoter and regulatory sequence with a constitutive SP01 phage promoter resulted in higher-level constitutive synthesis. Removal of a rho-independent terminator-like sequence located between the fifth (bioB) and sixth (bioI) genes prevented accumulation of the 5.1-kb transcript, suggesting that the putative terminator functions to limit expression of bioI, which is thought to be involved in an early step in biotin synthesis. PMID:8892842

  5. Identification and characterization of transcripts from the biotin biosynthetic operon of Bacillus subtilis.

    PubMed

    Perkins, J B; Bower, S; Howitt, C L; Yocum, R R; Pero, J

    1996-11-01

    Northern (RNA) blot analysis of the Bacillus subtilis biotin operon, bioWAFDBIorf2, detected at least two steady-state polycistronic transcripts initiated from a putative vegetative (Pbio) promoter that precedes the operon, i.e., a full-length 7.2-kb transcript covering the entire operon and a more abundant 5.1-kb transcript covering just the first five genes of the operon. Biotin and the B. subtilis birA gene product regulated synthesis of the transcripts. Moreover, replacing the putative Pbio promoter and regulatory sequence with a constitutive SP01 phage promoter resulted in higher-level constitutive synthesis. Removal of a rho-independent terminator-like sequence located between the fifth (bioB) and sixth (bioI) genes prevented accumulation of the 5.1-kb transcript, suggesting that the putative terminator functions to limit expression of bioI, which is thought to be involved in an early step in biotin synthesis.

  6. Development of a copula-based particle filter (CopPF) approach for hydrologic data assimilation under consideration of parameter interdependence

    NASA Astrophysics Data System (ADS)

    Fan, Y. R.; Huang, G. H.; Baetz, B. W.; Li, Y. P.; Huang, K.

    2017-06-01

    In this study, a copula-based particle filter (CopPF) approach was developed for sequential hydrological data assimilation by considering parameter correlation structures. In CopPF, multivariate copulas are proposed to reflect parameter interdependence before the resampling procedure with new particles then being sampled from the obtained copulas. Such a process can overcome both particle degeneration and sample impoverishment. The applicability of CopPF is illustrated with three case studies using a two-parameter simplified model and two conceptual hydrologic models. The results for the simplified model indicate that model parameters are highly correlated in the data assimilation process, suggesting a demand for full description of their dependence structure. Synthetic experiments on hydrologic data assimilation indicate that CopPF can rejuvenate particle evolution in large spaces and thus achieve good performances with low sample size scenarios. The applicability of CopPF is further illustrated through two real-case studies. It is shown that, compared with traditional particle filter (PF) and particle Markov chain Monte Carlo (PMCMC) approaches, the proposed method can provide more accurate results for both deterministic and probabilistic prediction with a sample size of 100. Furthermore, the sample size would not significantly influence the performance of CopPF. Also, the copula resampling approach dominates parameter evolution in CopPF, with more than 50% of particles sampled by copulas in most sample size scenarios.

  7. Molecular Characterization of Copper and Cadmium Resistance Determinants in the Biomining Thermoacidophilic Archaeon Sulfolobus metallicus

    PubMed Central

    Orell, Alvaro; Remonsellez, Francisco; Arancibia, Rafaela; Jerez, Carlos A.

    2013-01-01

    Sulfolobus metallicus is a thermoacidophilic crenarchaeon used in high-temperature bioleaching processes that is able to grow under stressing conditions such as high concentrations of heavy metals. Nevertheless, the genetic and biochemical mechanisms responsible for heavy metal resistance in S. metallicus remain uncharacterized. Proteomic analysis of S. metallicus cells exposed to 100 mM Cu revealed that 18 out of 30 upregulated proteins are related to the production and conversion of energy, amino acids biosynthesis, and stress responses. Ten of these last proteins were also up-regulated in S. metallicus treated in the presence of 1 mM Cd suggesting that at least in part, a common general response to these two heavy metals. The S. metallicus genome contained two complete cop gene clusters, each encoding a metallochaperone (CopM), a Cu-exporting ATPase (CopA), and a transcriptional regulator (CopT). Transcriptional expression analysis revealed that copM and copA from each cop gene cluster were cotranscribed and their transcript levels increased when S. metallicus was grown either in the presence of Cu or using chalcopyrite (CuFeS2) as oxidizable substrate. This study shows for the first time the presence of a duplicated version of the cop gene cluster in Archaea and characterizes some of the Cu and Cd resistance determinants in a thermophilic archaeon employed for industrial biomining. PMID:23509422

  8. Molecular characterization of copper and cadmium resistance determinants in the biomining thermoacidophilic archaeon Sulfolobus metallicus.

    PubMed

    Orell, Alvaro; Remonsellez, Francisco; Arancibia, Rafaela; Jerez, Carlos A

    2013-01-01

    Sulfolobus metallicus is a thermoacidophilic crenarchaeon used in high-temperature bioleaching processes that is able to grow under stressing conditions such as high concentrations of heavy metals. Nevertheless, the genetic and biochemical mechanisms responsible for heavy metal resistance in S. metallicus remain uncharacterized. Proteomic analysis of S. metallicus cells exposed to 100 mM Cu revealed that 18 out of 30 upregulated proteins are related to the production and conversion of energy, amino acids biosynthesis, and stress responses. Ten of these last proteins were also up-regulated in S. metallicus treated in the presence of 1 mM Cd suggesting that at least in part, a common general response to these two heavy metals. The S. metallicus genome contained two complete cop gene clusters, each encoding a metallochaperone (CopM), a Cu-exporting ATPase (CopA), and a transcriptional regulator (CopT). Transcriptional expression analysis revealed that copM and copA from each cop gene cluster were cotranscribed and their transcript levels increased when S. metallicus was grown either in the presence of Cu or using chalcopyrite (CuFeS2) as oxidizable substrate. This study shows for the first time the presence of a duplicated version of the cop gene cluster in Archaea and characterizes some of the Cu and Cd resistance determinants in a thermophilic archaeon employed for industrial biomining.

  9. Statistical analysis of dust signals observed by ROSINA/COPS onboard of the Rosetta spacecraft at comet 67P/Churyumov-Gerasimenko

    NASA Astrophysics Data System (ADS)

    Tzou, Chia-Yu; altwegg, kathrin; Bieler, Andre; Calmonte, Ursina; Gasc, Sébastien; Le Roy, Léna; Rubin, Martin

    2016-10-01

    ROSINA is the in situ Rosetta Orbiter Spectrometer for Ion and Neutral Analysis on board of Rosetta, one of the corner stone missions of the European Space Agency (ESA) to land and orbit the Jupiter family comet 67P/Churyumov-Gerasimenko (67P). ROSINA consists of two mass spectrometers and a pressure sensor. The Reflectron Time of Flight Spectrometer (RTOF) and the Double Focusing Mass Spectrometer (DFMS) complement each other in mass and time resolution.The Comet Pressure Sensor (COPS) provides density measurements of the neutral molecules in the cometary coma of 67P. COPS has two gauges, a nude gauge that measures the total neutral density and a ram gauge that measures the dynamic pressure from the comet. Combining the two COPS is also capable of providing gas dynamic information such as gas velocity and gas temperature of the coma.While Rosetta started orbiting around 67P in August 2014, COPS observed diurnal and seasonal variations of the neutral gas density in the coma. Surprisingly, additional to these major density variation patterns, COPS occasionally observed small spikes in the density that are associated with dust. These dust signals can be interpreted as a result of cometary dust releasing volatiles while heated up near COPS. A statistical analysis of dust signals detected by COPS will be presented.

  10. Augmented feedback of COM and COP modulates the regulation of quiet human standing relative to the stability boundary.

    PubMed

    Kilby, Melissa C; Slobounov, Semyon M; Newell, Karl M

    2016-06-01

    The experiment manipulated real-time kinematic feedback of the motion of the whole body center of mass (COM) and center of pressure (COP) in anterior-posterior (AP) and medial-lateral (ML) directions to investigate the variables actively controlled in quiet standing of young adults. The feedback reflected the current 2D postural positions within the 2D functional stability boundary that was scaled to 75%, 30% and 12% of its original size. The findings showed that the distance of both COP and COM to the respective stability boundary was greater during the feedback trials compared to a no feedback condition. However, the temporal safety margin of the COP, that is, the virtual time-to-contact (VTC), was higher without feedback. The coupling relation of COP-COM showed stable in-phase synchronization over all of the feedback conditions for frequencies below 1Hz. For higher frequencies (up to 5Hz), there was progressive reduction of COP-COM synchronization and local adaptation under the presence of augmented feedback. The findings show that the augmented feedback of COM and COP motion differentially and adaptively influences spatial and temporal properties of postural motion relative to the stability boundary while preserving the organization of the COM-COP coupling in postural control. Copyright © 2016. Published by Elsevier B.V.

  11. Knockout of an outer membrane protein operon of anaplasma marginale by transposon mutagenesis

    USDA-ARS?s Scientific Manuscript database

    Large amounts of data generated by genomics, transcriptomics and proteomics technologies have increased our understanding of the biology of Anaplasma marginale. However, these data have also led to new assumptions that require testing, ideally through classic genetic mutation. One example is the def...

  12. 30 CFR 585.811 - When must I follow my Safety Management System?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ..., COPs and GAPs Safety Management Systems § 585.811 When must I follow my Safety Management System? Your... COP, SAP, or GAP. You must conduct all activities described in your approved COP, SAP, or GAP in...

  13. 30 CFR 585.811 - When must I follow my Safety Management System?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ..., COPs and GAPs Safety Management Systems § 585.811 When must I follow my Safety Management System? Your... COP, SAP, or GAP. You must conduct all activities described in your approved COP, SAP, or GAP in...

  14. 30 CFR 585.811 - When must I follow my Safety Management System?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ..., COPs and GAPs Safety Management Systems § 585.811 When must I follow my Safety Management System? Your... COP, SAP, or GAP. You must conduct all activities described in your approved COP, SAP, or GAP in...

  15. mar Operon Involved in Multidrug Resistance of Enterobacter aerogenes

    PubMed Central

    Chollet, Renaud; Bollet, Claude; Chevalier, Jacqueline; Malléa, Monique; Pagès, Jean-Marie; Davin-Regli, Anne

    2002-01-01

    We determined the sequence of the entire marRAB operon in Enterobacter aerogenes. It is functionally and structurally analogous to the Escherichia coli operon. The overexpression of E. aerogenes MarA induces a multidrug resistance phenotype in a susceptible strain, demonstrated by a noticeable resistance to various antibiotics, a decrease in immunodetected porins, and active efflux of norfloxacin. PMID:11897595

  16. The Staphylococcus aureus group II biotin protein ligase BirA is an effective regulator of biotin operon transcription and requires the DNA binding domain for full enzymatic activity.

    PubMed

    Henke, Sarah K; Cronan, John E

    2016-11-01

    Group II biotin protein ligases (BPLs) are characterized by the presence of an N-terminal DNA binding domain that functions in transcriptional regulation of the genes of biotin biosynthesis and transport. The Staphylococcus aureus Group II BPL which is called BirA has been reported to bind an imperfect inverted repeat located upstream of the biotin synthesis operon. DNA binding by other Group II BPLs requires dimerization of the protein which is triggered by synthesis of biotinoyl-AMP (biotinoyl-adenylate), the intermediate in the ligation of biotin to its cognate target proteins. However, the S. aureus BirA was reported to dimerize and bind DNA in the absence of biotin or biotinoyl-AMP (Soares da Costa et al. (2014) Mol Microbiol 91: 110-120). These in vitro results argued that the protein would be unable to respond to the levels of biotin or acceptor proteins and thus would lack the regulatory properties of the other characterized BirA proteins. We tested the regulatory function of the protein using an in vivo model system and examined its DNA binding properties in vitro using electrophoretic mobility shift and fluorescence anisotropy analyses. We report that the S. aureus BirA is an effective regulator of biotin operon transcription and that the prior data can be attributed to artifacts of mobility shift analyses. We also report that deletion of the DNA binding domain of the S. aureus BirA results in loss of virtually all of its ligation activity. © 2016 John Wiley & Sons Ltd.

  17. COPS5 (Jab1) protein increases β site processing of amyloid precursor protein and amyloid β peptide generation by stabilizing RanBP9 protein levels.

    PubMed

    Wang, Hongjie; Dey, Debleena; Carrera, Ivan; Minond, Dmitriy; Bianchi, Elisabetta; Xu, Shaohua; Lakshmana, Madepalli K

    2013-09-13

    Increased processing of amyloid precursor protein (APP) and accumulation of neurotoxic amyloid β peptide (Aβ) in the brain is central to the pathogenesis of Alzheimer's disease (AD). Therefore, the identification of molecules that regulate Aβ generation is crucial for future therapeutic approaches for AD. We demonstrated previously that RanBP9 regulates Aβ generation in a number of cell lines and primary neuronal cultures by forming tripartite protein complexes with APP, low-density lipoprotein-related protein, and BACE1, consequently leading to increased amyloid plaque burden in the brain. RanBP9 is a scaffold protein that exists and functions in multiprotein complexes. To identify other proteins that may bind RanBP9 and regulate Aβ levels, we used a two-hybrid analysis against a human brain cDNA library and identified COPS5 as a novel RanBP9-interacting protein. This interaction was confirmed by coimmunoprecipitation experiments in both neuronal and non-neuronal cells and mouse brain. Colocalization of COPS5 and RanBP9 in the same subcellular compartments further supported the interaction of both proteins. Furthermore, like RanBP9, COPS5 robustly increased Aβ generation, followed by increased soluble APP-β (sAPP-β) and decreased soluble-APP-α (sAPP-α) levels. Most importantly, down-regulation of COPS5 by siRNAs reduced Aβ generation, implying that endogenous COPS5 regulates Aβ generation. Finally, COPS5 levels were increased significantly in AD brains and APΔE9 transgenic mice, and overexpression of COPS5 strongly increased RanBP9 protein levels by increasing its half-life. Taken together, these results suggest that COPS5 increases Aβ generation by increasing RanBP9 levels. Thus, COPS5 is a novel RanBP9-binding protein that increases APP processing and Aβ generation by stabilizing RanBP9 protein levels.

  18. A role for tubular networks and a COP I-independent pathway in the mitotic fragmentation of Golgi stacks in a cell-free system

    PubMed Central

    1995-01-01

    Golgi stacks were previously shown to be converted into tubular networks when incubated in mitotic cytosol depleted of the coatomer subunit of COP I coats (Misteli and Warren, 1994). Similar, though smaller, networks are now shown to be an early intermediate on the Golgi fragmentation pathway both in vitro and in vivo. Their appearance mirrors the disappearance of Golgi cisternae and at their peak they constitute 35% of total Golgi membrane. They are consumed by two pathways, the first involving the budding of COP I-coated vesicles described previously (Misteli and Warren, 1994). The second involves a COP I-independent mechanism that leads eventually to a vesicle fraction that is larger in size and more heterogeneous than that produced by the COP I-mechanism. We suggest that both pathways operate concurrently at the onset of mitotic fragmentation. The COP I-independent pathway converts cisternae into tubular networks that then fragment. The COP I- dependent pathway partially consumes first the cisternae at the beginning of the incubation and then the tubular networks that form from them. PMID:7657690

  19. Demographic and clinical characteristics of carbon monoxide poisoning: nationwide data between 1999 and 2012 in Taiwan.

    PubMed

    Huang, Chien-Cheng; Ho, Chung-Han; Chen, Yi-Chen; Lin, Hung-Jung; Hsu, Chien-Chin; Wang, Jhi-Joung; Su, Shih-Bin; Guo, How-Ran

    2017-07-14

    Carbon monoxide poisoning (COP) is not uncommon, but nationwide epidemiological data are limited. We conducted a study in Taiwan to fill the data gaps. We used a nationwide poisoning database to study all COP patients in Taiwan diagnosed between 1999 and 2012. We conducted descriptive analyses and compared the differences between the two sexes. In addition, we assessed the trends in suicide and mortality rates from 1999 to 2012. We identified 25,912 COP patients with an almost equal female and male distribution (50.6% vs. 49.4%). The mean age was 36.0 years, and most of the patients were between 20 and 50 years old. The highest incidence rate in the year occurred during winter. While female patients were more likely to have mental disorders (35.9% vs. 28.1%, p < 0.001), male patients were more likely to be resulted from suicide attempts (22.9% vs. 17.7%, p < 0.001). In both sexes, the suicidal rate increased from 1999, reached the peak in 2007, and then decreased gradually. Hyperbaric oxygen therapy was performed in 24.2% of the patients. Neurological sequelae developed in 9.1% of the patients, and chronic respiratory failure and requirement of long-term care were observed in 5.0% and 0.1% of the patients. The 1-month and 3-month mortality rates were 1.6% and 5.0%. The 3-month mortality rate did not show significant change between 1999 and 2012. This study showed a whole picture of COP in Taiwan, which could add to the important knowledge of this disastrous problem in public health. Some important findings, including higher percentages of mental disorders in female patients and suicide attempt in male patients, seasonal changes, and trends in mortality and morbidity (suicide) rates, may help developing strategies for prevention and treatment of COP.

  20. Relation between lowered colloid osmotic pressure, respiratory failure, and death.

    PubMed

    Tonnesen, A S; Gabel, J C; McLeavey, C A

    1977-01-01

    Plasma colloid osmotic pressure was measured each day in 84 intensive care unit patients. Probit analysis demonstrated a direct relationship between colloid osmotic pressure (COP) and survival. The COP associated with a 50% survival rate was 15.0 torr. COP was higher in survivors than in nonsurvivors without respiratory failure and in patients who recovered from respiratory failure. We conclude that lowered COP is associated with an elevated mortality rate. However, the relationship to death is not explained by the relationship to respiratory failure.

  1. CHOP or THP-COP regimens in the treatment of newly diagnosed peripheral T-cell lymphoma, not otherwise specified: a comparison of doxorubicin and pirarubicin.

    PubMed

    Shibata, Yuhei; Hara, Takeshi; Kasahara, Senji; Yamada, Toshiki; Sawada, Michio; Mabuchi, Ryoko; Matsumoto, Takuro; Nakamura, Nobuhiko; Nakamura, Hiroshi; Ninomiya, Soranobu; Kitagawa, Junichi; Kanemura, Nobuhiro; Kito, Yusuke; Goto, Naoe; Miyazaki, Tatsuhiko; Takami, Tsuyoshi; Takeuchi, Tamotsu; Shimizu, Masahito; Tsurumi, Hisashi

    2017-06-01

    The CHOP regimen consisting of cyclophosphamide, doxorubicin (DOX), vincristine and prednisolone has been the most used regimen for peripheral T-cell lymphoma, not otherwise specified (PTCL-NOS). Pirarubicin [tetrahydropyranyladriamycin (THP)], a derivative of DOX, is an anthracycline with reportedly less cardiotoxicity than DOX. Here, we confirmed the efficacy of THP-COP using THP instead of DOX in the treatment of PTCL-NOS. The study protocol employed a retrospective, consecutive entry design. We retrospectively analysed 56 patients with PTCL-NOS who had received THP-COP or CHOP. These regimens were performed every 21 days. Twenty-nine patients received THP-COP, and 27 received CHOP. There were no significant differences in known prognostic factors, including in the International Prognostic Index (IPI) and the prognostic index for T-cell lymphoma (PIT), between the two groups. Complete remission rates in patients with THP-COP and CHOP were 52% in both groups; the 3-year overall survival (OS) rates were 67% and 52% (p = 0.074), and the 3-year progression-free survival (PFS) rates were 51% and 29% (p = 0.070), respectively. In patients with low IPI (low or low-intermediate), THP-COP had significantly better 3-year OS (100% vs. 64%; p < 0.001) and 3-year PFS (75% vs. 33%; p < 0.05) than CHOP. Similar differences between THP-COP and CHOP were observed in patients with a low PIT (groups 1 or 2). Our study showed that THP-COP produced results equivalent to CHOP regarding efficacy and safety in patients with PTCL-NOS. In patients with low IPI or PIT, THP-COP resulted in significantly better prognosis. Copyright © 2015 John Wiley & Sons, Ltd. Copyright © 2015 John Wiley & Sons, Ltd.

  2. Increased risk for diabetes mellitus in patients with carbon monoxide poisoning

    PubMed Central

    Huang, Chien-Cheng; Ho, Chung-Han; Chen, Yi-Chen; Lin, Hung-Jung; Hsu, Chien-Chin; Wang, Jhi-Joung; Su, Shih-Bin; Guo, How-Ran

    2017-01-01

    Carbon monoxide poisoning (COP) causes hypoxic injury and inflammatory and immunological reactions in the brain and local organs including the pancreas. Therefore, it is plausible that COP may increase the risk for developing diabetes mellitus (DM), but studies on this possible association are limited. We conducted a nationwide study in Taiwan to fill the data gap. We used the Nationwide Poisoning Database and the Longitudinal Health Insurance Database 2000 to identify all COP patients diagnosed between 1999 and 2012 (the study cohort) and then construct a comparison cohort of patients without COP through matching at 1:3 by the index date and age. The risk for DM between the two cohorts was compared by following up until 2013. We also investigated the independent predictors for DM in all the patients. During the study period, 22,308 COP patients were identified, and 66,924 non-COP patients were included in the comparison cohort accordingly. Patients with COP had an increased risk for DM with an adjusted hazard ratio (AHR) of 1.92 (95% confidence interval [CI]: 1.79–2.06) after adjusting for age, sex, comorbidities, and monthly income, especially in the subgroups of age <35 years, age ≥ 65 years, female sex, and comorbidities with congestive heart failure, hyperthyroidism, and polycystic ovary syndrome. Cox proportional hazard regression analysis showed that the increased risk for DM was highest in the first month after COP (AHR= 3.38; 95% CI: 2.29–4.99) and lasted even after 4 years (AHR= 1.82; 95% CI: 1.62–2.04). We found that COP, older age, male sex, hypertension, hyperlipidemia, hyperuricemia, and low monthly income were independent predictors for DM. Intervention studies are needed to validate the results and delineate the detailed mechanisms. PMID:28969020

  3. A horse’s locomotor signature: COP path determined by the individual limb

    PubMed Central

    Hobbs, Sarah Jane; Back, Willem

    2017-01-01

    Introduction Ground reaction forces in sound horses with asymmetric hooves show systematic differences in the horizontal braking force and relative timing of break-over. The Center Of Pressure (COP) path quantifies the dynamic load distribution under the hoof in a moving horse. The objective was to test whether anatomical asymmetry, quantified by the difference in dorsal wall angle between the left and right forelimbs, correlates with asymmetry in the COP path between these limbs. In addition, repeatability of the COP path was investigated. Methods A larger group (n = 31) visually sound horses with various degree of dorsal hoof wall asymmetry trotted three times over a pressure mat. COP path was determined in a hoof-bound coordinate system. A relationship between correlations between left and right COP paths and degree of asymmetry was investigated. Results Using a hoof-bound coordinate system made the COP path highly repeatable and unique for each limb. The craniocaudal patterns are usually highly correlated between left and right, but the mediolateral patterns are not. Some patterns were found between COP path and dorsal wall angle but asymmetry in dorsal wall angle did not necessarily result in asymmetry in COP path and the same could be stated for symmetry. Conclusion This method is a highly sensitive method to quantify the net result of the interaction between all of the forces and torques that occur in the limb and its inertial properties. We argue that changes in motor control, muscle force, inertial properties, kinematics and kinetics can potentially be picked up at an early stage using this method and could therefore be used as an early detection method for changes in the musculoskeletal apparatus. PMID:28196073

  4. Interprofessional Communities of Practice in Continuing Medical Education for Promoting and Sustaining Practice Change: A Prospective Cohort Study.

    PubMed

    Barker, Megan; Lecce, Julia; Ivanova, Anna; Zawertailo, Laurie; Dragonetti, Rosa; Selby, Peter

    2018-01-01

    Standard knowledge delivery formats for CME may have limited impact on long-term practice change. A community of practice (CoP) is one tool that may enhance competencies and support practice change. This study explores the utility of an interprofessional CoP as an adjunct to a CME program in tobacco addiction treatment (Training Enhancement in Applied Counselling and Health [TEACH] Project) to promote and sustain practice change. A prospective cohort design was utilized to examine the long-term impact of the TEACH CoP on practice change. An online survey was administered to TEACH-trained practitioners to assess perceived feasibility, importance, and confidence related to course competencies, involvement in TEACH CoP activities, engagement in knowledge transfer (KT), and implementation of new programming. Chi-square tests were used to detect differences in KT and program development associated with CoP participation. Course competency scores from immediate postcourse surveys and long-term follow-up surveys were compared. No significant differences in participant characteristics were found between those who did (n = 300) and did not (n = 122) participate in the TEACH CoP. Mean self-perceived competency scores were greater immediately after course than at long-term follow-up; however, self-ratings of competency in pharmacological interventions and motivational interviewing were higher at follow-up. TEACH CoP participation was associated with significantly greater engagement in KT and implementation of new programming after training. The findings from this evaluation suggest the value of interprofessional CoPs offered posttraining as a mechanism to enhance practice. CME providers should consider offering CoPs as a component of training programs to promote and sustain practice change.

  5. Required coefficient of friction in the anteroposterior and mediolateral direction during turning at different walking speeds

    PubMed Central

    Yamaguchi, Takeshi; Suzuki, Akito; Hokkirigawa, Kazuo

    2017-01-01

    This study investigated the required coefficient of friction (RCOF) and the tangent of center of mass (COM)–center of pressure (COP) angle in the mediolateral (ML) and anteroposterior (AP) directions during turning at different walking speeds. Sixteen healthy young adults (8 males and 8 females) participated in this study. The participants were instructed to conduct trials of straight walking and 90° step and spin turns to the right at each of three self-selected speeds (slow, normal, and fast). The ML and AP directions during turning gait were defined using the orientation of the pelvis to construct a body-fixed reference frame. The RCOF values and COM–COP angle tangent in the ML direction during turning at weight acceptance phase were higher than those during straight walking, and those values increased with increasing walking speed. The ML component of the RCOF and COM–COP tangent values during weight acceptance for step turns were higher than those for spin turns. The mean centripetal force during turning tended to increase with an increase in walking speed and had a strong positive correlation with the RCOF values in the ML direction (R = 0.97 during the weight acceptance phase; R = 0.95 during the push-off phase). Therefore, turning, particularly step turn, is likely to cause lateral slip at weight acceptance because of the increased centripetal force compared with straight walking. Future work should test at-risk population and compare with the present results. PMID:28640853

  6. Required coefficient of friction in the anteroposterior and mediolateral direction during turning at different walking speeds.

    PubMed

    Yamaguchi, Takeshi; Suzuki, Akito; Hokkirigawa, Kazuo

    2017-01-01

    This study investigated the required coefficient of friction (RCOF) and the tangent of center of mass (COM)-center of pressure (COP) angle in the mediolateral (ML) and anteroposterior (AP) directions during turning at different walking speeds. Sixteen healthy young adults (8 males and 8 females) participated in this study. The participants were instructed to conduct trials of straight walking and 90° step and spin turns to the right at each of three self-selected speeds (slow, normal, and fast). The ML and AP directions during turning gait were defined using the orientation of the pelvis to construct a body-fixed reference frame. The RCOF values and COM-COP angle tangent in the ML direction during turning at weight acceptance phase were higher than those during straight walking, and those values increased with increasing walking speed. The ML component of the RCOF and COM-COP tangent values during weight acceptance for step turns were higher than those for spin turns. The mean centripetal force during turning tended to increase with an increase in walking speed and had a strong positive correlation with the RCOF values in the ML direction (R = 0.97 during the weight acceptance phase; R = 0.95 during the push-off phase). Therefore, turning, particularly step turn, is likely to cause lateral slip at weight acceptance because of the increased centripetal force compared with straight walking. Future work should test at-risk population and compare with the present results.

  7. An ABC Transporter System of Yersinia pestis Allows Utilization of Chelated Iron by Escherichia coli SAB11

    PubMed Central

    Bearden, Scott W.; Staggs, Teanna M.; Perry, Robert D.

    1998-01-01

    The acquisition of iron is an essential component in the pathogenesis of Yersinia pestis, the agent of bubonic and pneumonic plague. A cosmid library derived from the genomic DNA of Y. pestis KIM6+ was used for transduction of an Escherichia coli mutant (SAB11) defective in the biosynthesis of the siderophore enterobactin. Recombinant plasmids which had a common 13-kb BamHI fragment were isolated from SAB11 transductants in which growth but not enterobactin synthesis was restored on media containing the iron chelator EDDA [ethylenediamine-di(o-hydroxyphenyl acetic acid)]. Subcloning and transposon mutagenesis revealed a 5.6-kb region, designated yfe, essential for SAB11 growth stimulation. In vitro transcription-translation analysis identified polypeptides of 18, 29.5, 32, and 33 kDa encoded by the yfe locus. Sequence analysis shows this locus to be comprised of five genes in two separate operons which have potential Fur-binding sequences in both promoters. A putative polycistronic operon, yfeABCD, is Fur regulated and responds to iron and manganese. A functional Fur protein is required for the observed manganese repression of this operon. This operon encodes polypeptides which have strong similarity to the ATP-binding cassette (ABC) family of transporters and include a periplasmic binding protein (YfeA), an ATP-binding protein (YfeB), and two integral membrane proteins (YfeC and -D), which likely function in the acquisition of inorganic iron and possibly other ions. The ∼21-kDa protein encoded by the separately transcribed yfeE gene may be located in the cell envelope, since a yfeE::TnphoA fusion is PhoA+. Mutations in this gene abrogate growth of SAB11 on iron-chelated media. PMID:9495751

  8. An ABC transporter system of Yersinia pestis allows utilization of chelated iron by Escherichia coli SAB11.

    PubMed

    Bearden, S W; Staggs, T M; Perry, R D

    1998-03-01

    The acquisition of iron is an essential component in the pathogenesis of Yersinia pestis, the agent of bubonic and pneumonic plague. A cosmid library derived from the genomic DNA of Y. pestis KIM6+ was used for transduction of an Escherichia coli mutant (SAB11) defective in the biosynthesis of the siderophore enterobactin. Recombinant plasmids which had a common 13-kb BamHI fragment were isolated from SAB11 transductants in which growth but not enterobactin synthesis was restored on media containing the iron chelator EDDA [ethylenediamine-di(o-hydroxyphenyl acetic acid)]. Subcloning and transposon mutagenesis revealed a 5.6-kb region, designated yfe, essential for SAB11 growth stimulation. In vitro transcription-translation analysis identified polypeptides of 18, 29.5, 32, and 33 kDa encoded by the yfe locus. Sequence analysis shows this locus to be comprised of five genes in two separate operons which have potential Fur-binding sequences in both promoters. A putative polycistronic operon, yfeABCD, is Fur regulated and responds to iron and manganese. A functional Fur protein is required for the observed manganese repression of this operon. This operon encodes polypeptides which have strong similarity to the ATP-binding cassette (ABC) family of transporters and include a periplasmic binding protein (YfeA), an ATP-binding protein (YfeB), and two integral membrane proteins (YfeC and -D), which likely function in the acquisition of inorganic iron and possibly other ions. The approximately 21-kDa protein encoded by the separately transcribed yfeE gene may be located in the cell envelope, since a yfeE::TnphoA fusion is PhoA+. Mutations in this gene abrogate growth of SAB11 on iron-chelated media.

  9. 7 CFR 457.126 - Popcorn cop isurance povisions.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 6 2010-01-01 2010-01-01 false Popcorn cop isurance povisions. 457.126 Section 457.126 Agriculture Regulations of the Department of Agriculture (Continued) FEDERAL CROP INSURANCE CORPORATION, DEPARTMENT OF AGRICULTURE COMMON CROP INSURANCE REGULATIONS § 457.126 Popcorn cop isurance...

  10. Video game-based exercises for balance rehabilitation: a single-subject design.

    PubMed

    Betker, Aimee L; Szturm, Tony; Moussavi, Zahra K; Nett, Cristabel

    2006-08-01

    To investigate whether coupling foot center of pressure (COP)-controlled video games to standing balance exercises will improve dynamic balance control and to determine whether the motivational and challenging aspects of the video games would increase a subject's desire to perform the exercises and complete the rehabilitation process. Case study, pre- and postexercise. University hospital outpatient clinic. A young adult with excised cerebellar tumor, 1 middle-aged adult with single right cerebrovascular accident, and 1 middle-aged adult with traumatic brain injury. A COP-controlled, video game-based exercise system. The following were calculated during 12 different tasks: the number of falls, range of COP excursion, and COP path length. Postexercise, subjects exhibited a lower fall count, decreased COP excursion limits for some tasks, increased practice volume, and increased attention span during training. The COP-controlled video game-based exercise regime motivated subjects to increase their practice volume and attention span during training. This in turn improved subjects' dynamic balance control.

  11. Diminishing the self-stigma of mental illness by coming out proud.

    PubMed

    Corrigan, Patrick W; Larson, Jonathon E; Michaels, Patrick J; Buchholz, Blythe A; Rossi, Rachel Del; Fontecchio, Malia Javier; Castro, David; Gause, Michael; Krzyżanowski, Richard; Rüsch, Nicolas

    2015-09-30

    This randomized controlled trial examined the impact of the Coming Out Proud (COP) program on self-stigma, stigma stress, and depression. Research participants who experienced mental health challenges were randomly assigned to a three session COP program (n=51) or a waitlist control (n=75). Outcome measures that assessed the progressively harmful stages of self-stigma, stigma stress appraisals, and depression were administered at pre-test, post-test, and one-month follow-up. People completing COP showed significant improvement at post-test and follow-up in the more harmful aspects of self-stigma compared to the control group. COP participants also showed improvements in stigma stress appraisals. Women participating in COP showed significant post-test and follow-up reductions in depression after COP compared to the control group. Men did not show this effect. Future research should determine whether these benefits also enhance attitudes related to recovery, empowerment, and self-determination. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  12. Roles of CONSTITUTIVE PHOTOMORPHOGENIC 10 in Arabidopsis stomata development

    PubMed Central

    Delgado, Dolores; Ballesteros, Isabel; Mena, Montaña; Fenoll, Carmen

    2012-01-01

    Stomata are epidermal bi-celled structures that differentiate within special cell lineages initiated by a subset of protodermal cells. Recently, we showed that the Arabidopsis photomorphogenic repressor COP10 controls specific cell-lineage and cell-signaling developmental mechanisms in stomatal lineages. Loss-of-function cop10-1 mutant cotyledons and leaves produced (in the light and in the dark) abundant stomatal clusters, but nonlineage epidermal cells were not affected. Here we examine COP10 role in hypocotyls, cylindrical organs displaying a distinct epidermal organization with alternate files of protruding and non-protruding cells, with the latter producing a limited number of stomata. COP10 prevents stomatal clusters and restricts stomata production in hypocotyls; these roles are specific to lineage cells as in cotyledons, since COP10 loss of function does not elicit stomatal fate in nonlineage cells; COP10 also sustains the directional cell expansion of all hypocotyl epidermal cell types, and seems necessary for the differentiation between protruding and non-protruding cell files. PMID:22836493

  13. Transcription of the extended hyp-operon in Nostoc sp. strain PCC 7120

    PubMed Central

    Agervald, Åsa; Stensjö, Karin; Holmqvist, Marie; Lindblad, Peter

    2008-01-01

    Background The maturation of hydrogenases into active enzymes is a complex process and e.g. a correctly assembled active site requires the involvement of at least seven proteins, encoded by hypABCDEF and a hydrogenase specific protease, encoded either by hupW or hoxW. The N2-fixing cyanobacterium Nostoc sp. strain PCC 7120 may contain both an uptake and a bidirectional hydrogenase. The present study addresses the presence and expression of hyp-genes in Nostoc sp. strain PCC 7120. Results RT-PCRs demonstrated that the six hyp-genes together with one ORF may be transcribed as a single operon. Transcriptional start points (TSPs) were identified 280 bp upstream from hypF and 445 bp upstream of hypC, respectively, demonstrating the existence of several transcripts. In addition, five upstream ORFs located in between hupSL, encoding the small and large subunits of the uptake hydrogenase, and the hyp-operon, and two downstream ORFs from the hyp-genes were shown to be part of the same transcript unit. A third TSP was identified 45 bp upstream of asr0689, the first of five ORFs in this operon. The ORFs are annotated as encoding unknown proteins, with the exception of alr0692 which is identified as a NifU-like protein. Orthologues of the four ORFs asr0689-alr0692, with a highly conserved genomic arrangement positioned between hupSL, and the hyp genes are found in several other N2-fixing cyanobacteria, but are absent in non N2-fixing cyanobacteria with only the bidirectional hydrogenase. Short conserved sequences were found in six intergenic regions of the extended hyp-operon, appearing between 11 and 79 times in the genome. Conclusion This study demonstrated that five ORFs upstream of the hyp-gene cluster are co-transcribed with the hyp-genes, and identified three TSPs in the extended hyp-gene cluster in Nostoc sp. strain PCC 7120. This may indicate a function related to the assembly of a functional uptake hydrogenase, hypothetically in the assembly of the small subunit of the enzyme. PMID:18442387

  14. Role of P27 -P55 operon from Mycobacterium tuberculosis in the resistance to toxic compounds

    PubMed Central

    2011-01-01

    Background The P27-P55 (lprG-Rv1410c) operon is crucial for the survival of Mycobacterium tuberculosis, the causative agent of human tuberculosis, during infection in mice. P55 encodes an efflux pump that has been shown to provide Mycobacterium smegmatis and Mycobacterium bovis BCG with resistance to several drugs, while P27 encodes a mannosylated glycoprotein previously described as an antigen that modulates the immune response against mycobacteria. The objective of this study was to determine the individual contribution of the proteins encoded in the P27-P55 operon to the resistance to toxic compounds and to the cell wall integrity of M. tuberculosis. Method In order to test the susceptibility of a mutant of M. tuberculosis H37Rv in the P27-P55 operon to malachite green, sodium dodecyl sulfate, ethidium bromide, and first-line antituberculosis drugs, this strain together with the wild type strain and a set of complemented strains were cultivated in the presence and in the absence of these drugs. In addition, the malachite green decolorization rate of each strain was obtained from decolorization curves of malachite green in PBS containing bacterial suspensions. Results The mutant strain decolorized malachite green faster than the wild type strain and was hypersensitive to both malachite green and ethidium bromide, and more susceptible to the first-line antituberculosis drugs: isoniazid and ethambutol. The pump inhibitor reserpine reversed M. tuberculosis resistance to ethidium bromide. These results suggest that P27-P55 functions through an efflux-pump like mechanism. In addition, deletion of the P27-P55 operon made M. tuberculosis susceptible to sodium dodecyl sulfate, suggesting that the lack of both proteins causes alterations in the cell wall permeability of the bacterium. Importantly, both P27 and P55 are required to restore the wild type phenotypes in the mutant. Conclusions The results clearly indicate that P27 and P55 are functionally connected in processes that involve the preservation of the cell wall and the transport of toxic compounds away from the cells. PMID:21762531

  15. Correlates of protection for enteric vaccines.

    PubMed

    Holmgren, Jan; Parashar, Umesh D; Plotkin, Stanley; Louis, Jacques; Ng, Su-Peing; Desauziers, Eric; Picot, Valentina; Saadatian-Elahi, Mitra

    2017-06-08

    An immunological Correlate of Protection (CoP) is an immune response that is statistically interrelated with protection. Identification of CoPs for enteric vaccines would help design studies to improve vaccine performance of licensed vaccines in low income settings, and would facilitate the testing of future vaccines in development that might be more affordable. CoPs are lacking today for most existing and investigational enteric vaccines. In order to share the latest information on CoPs for enteric vaccines and to discuss novel approaches to correlate mucosal immune responses in humans with protection, the Foundation Mérieux organized an international conference of experts where potential CoPs for vaccines were examined using case-studies for both bacterial and viral enteric pathogens. Experts on the panel concluded that to date, all established enteric vaccine CoPs, such as those for hepatitis A, Vi typhoid and poliovirus vaccines, are based on serological immune responses even though these may poorly reflect the relevant gut immune responses or predict protective efficacy. Known CoPs for cholera, norovirus and rotavirus could be considered as acceptable for comparisons of similarly composed vaccines while more work is still needed to establish CoPs for the remaining enteric pathogens and their candidate vaccines. Novel approaches to correlate human mucosal immune responses with protection include the investigation of gut-originating antibody-secreting cells (ASCs), B memory cells and follicular helper T cells from samples of peripheral blood during their recirculation. Copyright © 2017.

  16. Cation-induced transcriptional regulation of the dlt operon of Staphylococcus aureus.

    PubMed

    Koprivnjak, Tomaz; Mlakar, Vid; Swanson, Lindsey; Fournier, Benedicte; Peschel, Andreas; Weiss, Jerrold P

    2006-05-01

    Lipoteichoic and wall teichoic acids (TA) are highly anionic cell envelope-associated polymers containing repeating polyglycerol/ribitol phosphate moieties. Substitution of TA with D-alanine is important for modulation of many cell envelope-dependent processes, such as activity of autolytic enzymes, binding of divalent cations, and susceptibility to innate host defenses. D-Alanylation of TA is diminished when bacteria are grown in medium containing increased NaCl concentrations, but the effects of increased salt concentration on expression of the dlt operon encoding proteins mediating D-alanylation of TA are unknown. We demonstrate that Staphylococcus aureus transcriptionally represses dlt expression in response to high concentrations of Na(+) and moderate concentrations of Mg(2+) and Ca(2+) but not sucrose. Changes in dlt mRNA are induced within 15 min and sustained for several generations of growth. Mg(2+)-induced dlt repression depends on the ArlSR two-component system. Northern blotting, reverse transcription-PCR, and SMART-RACE analyses suggest that the dlt transcript begins 250 bp upstream of the dltA start codon and includes an open reading frame immediately upstream of dltA. Chloramphenicol transacetylase transcriptional fusions indicate that a region encompassing the 171 to 325 bp upstream of dltA is required for expression and Mg(2+)-induced repression of the dlt operon in S. aureus.

  17. Cation-Induced Transcriptional Regulation of the dlt Operon of Staphylococcus aureus

    PubMed Central

    Koprivnjak, Tomaz; Mlakar, Vid; Swanson, Lindsey; Fournier, Benedicte; Peschel, Andreas; Weiss, Jerrold P.

    2006-01-01

    Lipoteichoic and wall teichoic acids (TA) are highly anionic cell envelope-associated polymers containing repeating polyglycerol/ribitol phosphate moieties. Substitution of TA with d-alanine is important for modulation of many cell envelope-dependent processes, such as activity of autolytic enzymes, binding of divalent cations, and susceptibility to innate host defenses. d-Alanylation of TA is diminished when bacteria are grown in medium containing increased NaCl concentrations, but the effects of increased salt concentration on expression of the dlt operon encoding proteins mediating d-alanylation of TA are unknown. We demonstrate that Staphylococcus aureus transcriptionally represses dlt expression in response to high concentrations of Na+ and moderate concentrations of Mg2+ and Ca2+ but not sucrose. Changes in dlt mRNA are induced within 15 min and sustained for several generations of growth. Mg2+-induced dlt repression depends on the ArlSR two-component system. Northern blotting, reverse transcription-PCR, and SMART-RACE analyses suggest that the dlt transcript begins 250 bp upstream of the dltA start codon and includes an open reading frame immediately upstream of dltA. Chloramphenicol transacetylase transcriptional fusions indicate that a region encompassing the 171 to 325 bp upstream of dltA is required for expression and Mg2+-induced repression of the dlt operon in S. aureus. PMID:16672616

  18. Agrobacterium tumefaciens can obtain sulphur from an opine that is synthesized by octopine synthase using S-methylmethionine as a substrate.

    PubMed

    Flores-Mireles, Ana Lidia; Eberhard, Anatol; Winans, Stephen C

    2012-06-01

    Agrobacterium tumefaciens incites plant tumours that produce nutrients called opines, which are utilized by the bacteria during host colonization. Various opines provide sources of carbon, nitrogen and phosphorous, but virtually nothing was previously known about how A. tumefaciens acquires sulphur during colonization. Some strains encode an operon required for the catabolism of the opine octopine. This operon contains a gene, msh, that is predicted to direct the conversion of S-methylmethionine (SMM) and homocysteine (HCys) to two equivalents of methionine. Purified Msh carried out this reaction, suggesting that SMM could be an intermediate in opine catabolism. Purified octopine synthase (Ocs, normally expressed in plant tumours) utilized SMM and pyruvate to produce a novel opine, designated sulfonopine, whose catabolism by the bacteria would regenerate SMM. Sulfonopine was produced by tobacco and Arabidopsis when colonized by A. tumefaciens and was utilized as sole source of sulphur by A. tumefaciens. Purified Ocs also used 13 other proteogenic and non-proteogenic amino acids as substrates, including three that contain sulphur. Sulfonopine and 11 other opines were tested for induction of octopine catabolic operon and all were able to do so. This is the first study of the acquisition of sulphur, an essential element, by this pathogen. © 2012 Blackwell Publishing Ltd.

  19. Structural and Biochemical Characterization of Gentisate 1,2-Dioxygenase from Escherichia coli O157:H7.

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

    Adams,M.; Singh, V.; Keller, B.

    2006-01-01

    Gentisic acid (2,5-dihydroxybenzoic acid) is a key intermediate in aerobic bacterial pathways that are responsible for the metabolism of a large number of aromatic compounds. The critical step of these pathways is the oxygen-dependent reaction catalysed by gentisate 1,2-dioxygenase which opens the aromatic ring of gentisate to form maleylpyruvate. From gentisic acid, the cell derives carbon and energy through the conversion of maleylpyruvate to central metabolites. We have confirmed the annotation of a gentisate 1,2-dioygenase from the pathogenic O157:H7 Escherichia coli strain and present the first structural characterization of this family of enzymes. The identity of the reaction product wasmore » revealed using tandem mass spectroscopy. The operon responsible for the degradation of gentisate in this organism exhibits a high degree of conservation with the gentisate-degrading operons of other pathogenic bacteria, including the Shiga toxin-producing E. coli O103:H2, but does not appear to be present in non-pathogenic strains. The acquisition of the gentisate operon may represent a special adaptation to meet carbon source requirements under conditions of environmental stress and may provide a selective advantage for enterohaemorrhagic E. coli relative to their non-pathogenic counterparts.« less

  20. 32 CFR 635.6 - Name checks.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... with AR 340-21. (b) Checks will be accomplished by a review of the COPS MPRS. Information will be... agencies for counterintelligence purposes. (d) COPS MPRS is a database, which will contain all military police reports filed worldwide. Authorized users of COPS MPRS can conduct name checks for criminal...

  1. 32 CFR 635.6 - Name checks.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... with AR 340-21. (b) Checks will be accomplished by a review of the COPS MPRS. Information will be... agencies for counterintelligence purposes. (d) COPS MPRS is a database, which will contain all military police reports filed worldwide. Authorized users of COPS MPRS can conduct name checks for criminal...

  2. 32 CFR 635.6 - Name checks.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... with AR 340-21. (b) Checks will be accomplished by a review of the COPS MPRS. Information will be... agencies for counterintelligence purposes. (d) COPS MPRS is a database, which will contain all military police reports filed worldwide. Authorized users of COPS MPRS can conduct name checks for criminal...

  3. 78 FR 7812 - Agency Information Collection Activities; Proposed new Collection; Comments Requested: COPS...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-02-04

    ... DEPARTMENT OF JUSTICE [OMB Number 1103-NEW] Agency Information Collection Activities; Proposed new Collection; Comments Requested: COPS Survey on Police Consolidation and Shared Services ACTION: 60-Day Notice. The Department of Justice (DOJ) Office of Community Oriented Policing Services (COPS) will be...

  4. 78 FR 20694 - Agency Information Collection Activities; Proposed New Collection; Comments Requested: COPS...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-04-05

    ... DEPARTMENT OF JUSTICE [OMB Number 1103-NEW] Agency Information Collection Activities; Proposed New Collection; Comments Requested: COPS Survey on Police Consolidation and Shared Services ACTION: 30-Day Notice. The Department of Justice (DOJ) Office of Community Oriented Policing Services (COPS) will be...

  5. Selecting and designing with the right thermoplastic polymer for your microfluidic chip: a close look into cyclo-olefin polymer

    NASA Astrophysics Data System (ADS)

    Nevitt, Mark

    2013-03-01

    Engineers who are developing microfluidic devices and bioMEMs for life science applications have many aspects to consider when selecting the proper base materials for constructing a device. While glass and polydimethylsiloxane (PDMS) are the staple materials for proof-of-concept and prototype chip fabrication, they are not a feasible solution for commercial production due to their slow, labor-intensive production rate. Alternatively, a molded or extruded thermoplastic solution can deliver the precision, consistency, and high volume capability required for commercial scale production. Traditional thermoplastics, such as polymethylmethacrylate (PMMA), polycarbonate (PC), and polystyrene (PS), are well known by development engineers in the bioscience community; however, cyclo-olefin polymer (COP), a relative newcomer in the world of plastics, is gaining increasing attention for use in microfluidic devices due to its unique balance of key properties compared to conventional thermoplastics. In this paper, we provide a comprehensive look at the properties which make COP an excellent candidate for providing the flow cell support and reagent storage functions in microfluidic assays. We also explore the processing attributes and capabilities of COP resin and film which are crucial for manufacturing high-performance microfluidic devices.

  6. Changes in postural sway as a consequence of wearing a military backpack.

    PubMed

    Heller, Michelle F; Challis, John H; Sharkey, Neil A

    2009-07-01

    Military personnel are often required to carry all of their personal supplies and equipment for long distances during both training and combat situations, creating many biomechanical and postural challenges for these individuals. In addition to other problems such as generalized fatigue and the development of stress fractures, significant external loads may also affect a soldier's postural sway. The purpose of this study was to assess changes in postural sway as a consequence of wearing a military backpack in females. Forty-three female subjects between the ages of 18 and 25 volunteered to participate. There were two conditions: unloaded and while wearing an 18.1 kg military backpack. Each subject stood with two feet on a force platform for 30s under both conditions while center of pressure (COP) data were collected. COP path length increased 64%, medial-lateral excursion increased 131%, anterior-posterior excursion increased 54%, and COP area increased 229% with addition of the backpack (p<0.0001 for all of these measures). These data show that wearing 18.1 kg of external weight in a military backpack increases the postural sway of females, which may in turn increase the likelihood of falls and injury.

  7. CalR is required for the expression of T6SS2 and the adhesion of Vibrio parahaemolyticus to HeLa cells.

    PubMed

    Zhang, Lingyu; Osei-Adjei, George; Zhang, Ying; Gao, He; Yang, Wenhui; Zhou, Dongsheng; Huang, Xinxiang; Yang, Huiying; Zhang, Yiquan

    2017-08-01

    Vibrio parahaemolyticus expresses one major virulence determinant T6SS2, which is constituted into three putative operons, i.e., VPA1027-1024, VPA1043-1028, and VPA1044-1046. CalR, a LysR-type transcriptional regulator, was originally identified as a repressor of the swarming motility and T3SS1 gene expression. As shown in this study, CalR binds to the promoter-proximal region of each of the three operons to activate their transcription, and moreover, CalR activates the adhesion of V. parahaemolyticus to HeLa cells. In addition, competitive EMSAs demonstrated that CalR acts as an antagonist of H-NS in V. parahaemolyticus. Collectively, these studies confirmed a new physiological role for CalR in V. parahaemolyticus.

  8. The Neuro-Mechanical Processes That Underlie Goal-Directed Medio-Lateral APA during Gait Initiation.

    PubMed

    Honeine, Jean-Louis; Schieppati, Marco; Crisafulli, Oscar; Do, Manh-Cuong

    2016-01-01

    Gait initiation (GI) involves passing from bipedal to unipedal stance. It requires a rapid movement of the center of foot pressure (CoP) towards the future swing foot and of the center of mass (CoM) in the direction of the stance foot prior to the incoming step. This anticipatory postural adjustment (APA) allows disengaging the swing leg from the ground and establishing favorable conditions for stepping. This study aimed to describe the neuro-mechanical process that underlies the goal-directed medio-lateral (ML) APA. We hypothesized that controlled knee flexion of the stance leg contributes to the initial ML displacement of the CoP and to the calibration of the first step. Fourteen subjects initiated gait starting from three different initial stance widths of 15 cm (Small), 30 cm (Medium), and 45 cm (Large). Optoelectronic, force platform and electromyogram (EMG) measurements were performed. During APA, soleus activity diminished bilaterally, while tibialis anterior (TA) activity increased, more so in the stance leg than in the swing leg, and to a larger extent with increasing initial stance width. Knee flexion of the stance leg was observed during APA and correlated with the ML CoP displacement towards the swing leg. ML CoP and CoM displacements during APA increased with increasing stance width. The activity of stance-leg TA was correlated with the degree of knee flexion. Swing-leg tensor fasciae latae (TFL) was also active during APA. Across subjects, when stance-leg tibialis activity was low, TFL activity was large and vice versa. The modulation of the ML CoP position during APA allowed the gravity-driven torque to place the CoM just lateral to the stance foot during step execution. Accordingly, the gravity-driven torque, the ML CoM velocity during step execution, and the step width at foot contact (FC) were lower in the Small and greater in the Large condition. Consequently, the position of the stepping foot at FC remained close to the sagittal plane in all three conditions. Conclusively, coordinated activation of hip abductors and ankle dorsiflexors during APA displaces the CoP towards the swing leg, and sets the contact position for the swing foot.

  9. The Neuro-Mechanical Processes That Underlie Goal-Directed Medio-Lateral APA during Gait Initiation

    PubMed Central

    Honeine, Jean-Louis; Schieppati, Marco; Crisafulli, Oscar; Do, Manh-Cuong

    2016-01-01

    Gait initiation (GI) involves passing from bipedal to unipedal stance. It requires a rapid movement of the center of foot pressure (CoP) towards the future swing foot and of the center of mass (CoM) in the direction of the stance foot prior to the incoming step. This anticipatory postural adjustment (APA) allows disengaging the swing leg from the ground and establishing favorable conditions for stepping. This study aimed to describe the neuro-mechanical process that underlies the goal-directed medio-lateral (ML) APA. We hypothesized that controlled knee flexion of the stance leg contributes to the initial ML displacement of the CoP and to the calibration of the first step. Fourteen subjects initiated gait starting from three different initial stance widths of 15 cm (Small), 30 cm (Medium), and 45 cm (Large). Optoelectronic, force platform and electromyogram (EMG) measurements were performed. During APA, soleus activity diminished bilaterally, while tibialis anterior (TA) activity increased, more so in the stance leg than in the swing leg, and to a larger extent with increasing initial stance width. Knee flexion of the stance leg was observed during APA and correlated with the ML CoP displacement towards the swing leg. ML CoP and CoM displacements during APA increased with increasing stance width. The activity of stance-leg TA was correlated with the degree of knee flexion. Swing-leg tensor fasciae latae (TFL) was also active during APA. Across subjects, when stance-leg tibialis activity was low, TFL activity was large and vice versa. The modulation of the ML CoP position during APA allowed the gravity-driven torque to place the CoM just lateral to the stance foot during step execution. Accordingly, the gravity-driven torque, the ML CoM velocity during step execution, and the step width at foot contact (FC) were lower in the Small and greater in the Large condition. Consequently, the position of the stepping foot at FC remained close to the sagittal plane in all three conditions. Conclusively, coordinated activation of hip abductors and ankle dorsiflexors during APA displaces the CoP towards the swing leg, and sets the contact position for the swing foot. PMID:27642280

  10. Parallel Evolution and Horizontal Gene Transfer of the pst Operon in Firmicutes from Oligotrophic Environments

    PubMed Central

    Moreno-Letelier, Alejandra; Olmedo, Gabriela; Eguiarte, Luis E.; Martinez-Castilla, Leon; Souza, Valeria

    2011-01-01

    The high affinity phosphate transport system (pst) is crucial for phosphate uptake in oligotrophic environments. Cuatro Cienegas Basin (CCB) has extremely low P levels and its endemic Bacillus are closely related to oligotrophic marine Firmicutes. Thus, we expected the pst operon of CCB to share the same evolutionary history and protein similarity to marine Firmicutes. Orthologs of the pst operon were searched in 55 genomes of Firmicutes and 13 outgroups. Phylogenetic reconstructions were performed for the pst operon and 14 concatenated housekeeping genes using maximum likelihood methods. Conserved domains and 3D structures of the phosphate-binding protein (PstS) were also analyzed. The pst operon of Firmicutes shows two highly divergent clades with no correlation to the type of habitat nor a phylogenetic congruence, suggesting horizontal gene transfer. Despite sequence divergence, the PstS protein had a similar 3D structure, which could be due to parallel evolution after horizontal gene transfer events. PMID:21461370

  11. Decaffeination and measurement of caffeine content by addicted Escherichia coli with a refactored N-demethylation operon from Pseudomonas putida CBB5.

    PubMed

    Quandt, Erik M; Hammerling, Michael J; Summers, Ryan M; Otoupal, Peter B; Slater, Ben; Alnahhas, Razan N; Dasgupta, Aurko; Bachman, James L; Subramanian, Mani V; Barrick, Jeffrey E

    2013-06-21

    The widespread use of caffeine (1,3,7-trimethylxanthine) and other methylxanthines in beverages and pharmaceuticals has led to significant environmental pollution. We have developed a portable caffeine degradation operon by refactoring the alkylxanthine degradation (Alx) gene cluster from Pseudomonas putida CBB5 to function in Escherichia coli. In the process, we discovered that adding a glutathione S-transferase from Janthinobacterium sp. Marseille was necessary to achieve N 7 -demethylation activity. E. coli cells with the synthetic operon degrade caffeine to the guanine precursor, xanthine. Cells deficient in de novo guanine biosynthesis that contain the refactored operon are ″addicted″ to caffeine: their growth density is limited by the availability of caffeine or other xanthines. We show that the addicted strain can be used as a biosensor to measure the caffeine content of common beverages. The synthetic N-demethylation operon could be useful for reclaiming nutrient-rich byproducts of coffee bean processing and for the cost-effective bioproduction of methylxanthine drugs.

  12. 7 CFR 457.126 - Popcorn cop isurance povisions.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 6 2011-01-01 2011-01-01 false Popcorn cop isurance povisions. 457.126 Section 457... CORPORATION, DEPARTMENT OF AGRICULTURE COMMON CROP INSURANCE REGULATIONS § 457.126 Popcorn cop isurance povisions. The Popcorn Crop Insurance Provisions for the 1999 and succeeding crop years are as follows: FCIC...

  13. 32 CFR 635.16 - General.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... the COPS MPRS and a systems administrator to ensure that the system is properly functioning. Reporting... System (DIBRS). The Army inputs its data into DIBRS utilizing COPS. Any data reported to DIBRS is only as good as the data reported into COPS, so the need for accuracy in reporting incidents and utilizing...

  14. 32 CFR 635.16 - General.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... the COPS MPRS and a systems administrator to ensure that the system is properly functioning. Reporting... System (DIBRS). The Army inputs its data into DIBRS utilizing COPS. Any data reported to DIBRS is only as good as the data reported into COPS, so the need for accuracy in reporting incidents and utilizing...

  15. 32 CFR 635.16 - General.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... the COPS MPRS and a systems administrator to ensure that the system is properly functioning. Reporting... System (DIBRS). The Army inputs its data into DIBRS utilizing COPS. Any data reported to DIBRS is only as good as the data reported into COPS, so the need for accuracy in reporting incidents and utilizing...

  16. 32 CFR 635.16 - General.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... the COPS MPRS and a systems administrator to ensure that the system is properly functioning. Reporting... System (DIBRS). The Army inputs its data into DIBRS utilizing COPS. Any data reported to DIBRS is only as good as the data reported into COPS, so the need for accuracy in reporting incidents and utilizing...

  17. 30 CFR 585.907 - How will BOEM process my decommissioning application?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ..., COPs and GAPs Decommissioning Applications § 585.907 How will BOEM process my decommissioning... your decommissioning application with the decommissioning general concept in your approved SAP, COP, or... revise your SAP, COP, or GAP, and BOEM will begin the appropriate NEPA analysis and other regulatory...

  18. 77 FR 63293 - Proposed Information Collection; Comment Request; Coastal Ocean Program Grants Proposal...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-16

    ... Oceanic and Atmospheric Administration's Coastal Ocean Program (COP) provides direct financial assistance.... The statutory authority for COP is Public Law 102-567 Section 201 (Coastal Ocean Program). In addition... to file annual progress reports and a project final report using COP formats. All of these...

  19. 76 FR 10065 - Agency Information Collection Activities: Revision of a Previously Approved Collection, With...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-02-23

    ... Information Collection Under Review: COPS Application Package. The Department of Justice (DOJ) Office of Community Oriented Policing Services (COPS), will be submitting the following information collection request.../Collection: COPS Application Package. (3) Agency form number, if any, and the applicable component of the...

  20. 32 CFR 635.24 - Updating the COPS MPRS.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 32 National Defense 4 2011-07-01 2011-07-01 false Updating the COPS MPRS. 635.24 Section 635.24 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY (CONTINUED) LAW ENFORCEMENT AND... ensure that every founded offense is reported into the COPS MPRS. Timely and accurate reporting is...

  1. 32 CFR 635.24 - Updating the COPS MPRS.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 32 National Defense 4 2010-07-01 2010-07-01 true Updating the COPS MPRS. 635.24 Section 635.24 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY (CONTINUED) LAW ENFORCEMENT AND... ensure that every founded offense is reported into the COPS MPRS. Timely and accurate reporting is...

  2. Flight Test of Advanced Digital Control Concepts.

    DTIC Science & Technology

    1982-03-01

    34 equations, and this result is then compared against the "model" equations. OPTION :::CREATrE ,CIEY, COFPY, MAT ,AMAT, COIP-YUMAT, BMAT , 74 COPFY, CMAT...BMATY .-COPY, NMAT, AMAT ,72 ,COPY, GMAT, RMA, COP:Y, (MAT, BMA’TCOFY, MA T ,AMAT >74, COPY, CMA’TsmA, PCOPY, RMAT, BmAT , pCOPYVMiAT , AMAo.T 72...COP’Y, MAT, AMAT, >75, COPY , MAtT, BMAT , COPY, ZMAT, AMAT, 74 ,COP’Y, MAT, TMAT, COPY, VMAT, AMAT, .:COPY ,RMAiT, MAT,73,COPW GMAT, E4MAT, COP-"YTMAT

  3. MdCOP1 Ubiquitin E3 Ligases Interact with MdMYB1 to Regulate Light-Induced Anthocyanin Biosynthesis and Red Fruit Coloration in Apple1[W][OA

    PubMed Central

    Li, Yuan-Yuan; Mao, Ke; Zhao, Cheng; Zhao, Xian-Yan; Zhang, Hua-Lei; Shu, Huai-Rui; Hao, Yu-Jin

    2012-01-01

    MdMYB1 is a crucial regulator of light-induced anthocyanin biosynthesis and fruit coloration in apple (Malus domestica). In this study, it was found that MdMYB1 protein accumulated in the light but degraded via a ubiquitin-dependent pathway in the dark. Subsequently, the MdCOP1-1 and MdCOP1-2 genes were isolated from apple fruit peel and were functionally characterized in the Arabidopsis (Arabidopsis thaliana) cop1-4 mutant. Yeast (Saccharomyces cerevisiae) two-hybrid, bimolecular fluorescence complementation, and coimmunoprecipitation assays showed that MdMYB1 interacts with the MdCOP1 proteins. Furthermore, in vitro and in vivo experiments indicated that MdCOP1s are necessary for the ubiquitination and degradation of MdMYB1 protein in the dark and are therefore involved in the light-controlled stability of the MdMYB1 protein. Finally, a viral vector-based transformation approach demonstrated that MdCOP1s negatively regulate the peel coloration of apple fruits by modulating the degradation of the MdMYB1 protein. Our findings provide new insight into the mechanism by which light controls anthocyanin accumulation and red fruit coloration in apple and even other plant species. PMID:22855936

  4. Improvement of the COP of the LiBr-Water Double-Effect Absorption Cycles

    NASA Astrophysics Data System (ADS)

    Shitara, Atsushi

    Prevention of the global warming has called for a great necessity for energy saving. This applies to the improvement of the COP of absorption chiller-heaters. We started the development of the high efficiency gas-fired double-effect absorption chiller-heater using LiBr-H2O to achieve target performance in short or middle term. To maintain marketability, the volume of the high efficiency machine has been set below the equal to the conventional machine. The absorption cycle technology for improving the COP and the element technology for downsizing the machine is necessary in this development. In this study, the former is investigated. In this report, first of all the target performance has been set at cooling COP of 1.35(on HHV), which is 0.35 higher than the COP of 1.0 for conventional machines in the market. This COP of 1.35 is practically close to the maximum limit achievable by double-effect absorption chiller-heater. Next, the design condition of each element to achieve the target performance and the effect of each mean to improve the COP are investigated. Moreover, as a result of comparing the various flows(series, parallel, reverse)to which the each mean is applied, it has been found the optimum cycle is the parallel flow.

  5. Evidence for involvement of the C-terminal domain in the dimerization of the CopY repressor protein from Enterococcus hirae

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

    Pazehoski, Kristina O., E-mail: pazehosk@pitt.edu; Cobine, Paul A., E-mail: pac0006@auburn.edu; Winzor, Donald J.

    2011-03-11

    Research highlights: {yields} A metal-binding protein domain is directly involved in protein dimerization. {yields} Fusing the metal-binding domain to a monomeric protein induces dimerization. {yields} Frontal size-exclusion chromatography measures the strength of dimer interaction. {yields} Ultracentrifugation studies confirm the influence of metal binding on dimerization. -- Abstract: Metal binding to the C-terminal region of the copper-responsive repressor protein CopY is responsible for homodimerization and the regulation of the copper homeostasis pathway in Enterococcus hirae. Specific involvement of the 38 C-terminal residues of CopY in dimerization is indicated by zonal and frontal (large zone) size-exclusion chromatography studies. The studies demonstrate thatmore » the attachment of these CopY residues to the immunoglobulin-binding domain of streptococcal protein G (GB1) promotes dimerization of the monomeric protein. Although sensitivity of dimerization to removal of metal from the fusion protein is smaller than that found for CopY (as measured by ultracentrifugation studies), the demonstration that an unrelated protein (GB1) can be induced to dimerize by extending its sequence with the C-terminal portion of CopY confirms the involvement of this region in CopY homodimerization.« less

  6. Predictors of carbon monoxide poisoning-induced delayed neuropsychological sequelae.

    PubMed

    Ku, Hsiao-Lun; Yang, Kai-Chun; Lee, Ying-Chiao; Lee, Ming-Been; Chou, Yuan-Hwa

    2010-01-01

    Carbon monoxide poisoning (COP) commonly results in delayed neuropsychological sequelae (DNS). The aim of the article is to demonstrate the clinical characteristics and potential predictors of COP-induced DNS later. Retrospective medical record review was performed for patients who had COP in the past year at a National Medical Center in Taiwan. Sixty patients with COP were registered during a one-year period. Fifty-six of them (93.3%) were COP because of suicide attempt. Patients with COP who have a complete medical record of carboxyhemoglobin (COHb) and Glasgow Coma Scale (GCS) and Mini-Mental Status Examination (MMSE) scores were recruited. Multiple regression analysis was performed to search for the predictive factors of DNS. Forty-three patients were recruited. Most had attempted suicide (93.0%) using CO, and thirteen developed DNS later. A longer duration of admission, more sessions of hyperbaric oxygen therapy, and positive findings in brain computed tomography (CT) scans were more often found in patients with DNS than those without DNS. The GCS and MMSE scores and positive findings in brain CT scans were associated with the development of DNS but COHb was not. Our results identified several potential predictors of DNS. This finding may help clinicians understand and treat COP patients efficiently. Copyright 2010 Elsevier Inc. All rights reserved.

  7. Performance characteristics of low global warming potential R134a alternative refrigerants in ejector-expansion refrigeration system

    NASA Astrophysics Data System (ADS)

    Mishra, Shubham; Sarkar, Jahar

    2016-12-01

    Performance assessment of ejector-expansion vapor compression refrigeration system with eco-friendly R134a alternative refrigerants (R152a, R1234yf, R600a, R600, R290, R161, R32, and propylene) is presented for air-conditioning application. Ejector has been modeled by considering experimental data based correlations of component efficiencies to take care of all irreversibilities. Ejector area ratio has been optimized based on maximum coefficient of performance (COP) for typical air-conditioner operating temperatures. Selected refrigerants have been compared based on area ratio, pressure lift ratio, entrainment ratio, COP, COP improvement and volumetric cooling capacity. Effects of normal boiling point and critical point on the performances have been studied as well. Using ejector as an expansion device, maximum improvement in COP is noted in R1234yf (10.1%), which reduces the COP deviation with R134a (4.5% less in basic cycle and 2.5% less in ejector cycle). Hence, R1234yf seems to be best alternative for ejector expansion system due to its mild flammability and comparable volumetric capacity and cooling COP. refrigerant R161 is superior to R134a in terms of both COP and volumetric cooling capacity, although may be restricted for low capacity application due to its flammability.

  8. Single leg balancing in ballet: effects of shoe conditions and poses.

    PubMed

    Lobo da Costa, Paula H; Azevedo Nora, Fernanda G S; Vieira, Marcus Fraga; Bosch, Kerstin; Rosenbaum, Dieter

    2013-03-01

    The purpose of this study was to describe the effects of lower limb positioning and shoe conditions on stability levels of selected single leg ballet poses performed in demi-pointe position. Fourteen female non-professional ballet dancers (mean age of 18.4±2.8 years and mean body mass index of 21.5±2.8kg/m(2)) who had practiced ballet for at least seven years, without any musculoskeletal impairment volunteered to participate in this study. A capacitive pressure platform allowed for the assessment of center of pressure variables related to the execution of three single leg ballet poses in demi pointé position: attitude devant, attitude derriére, and attitude a la second. Peak pressures, contact areas, COP oscillation areas, anterior-posterior and medio-lateral COP oscillations and velocities were compared between two shoe conditions (barefoot versus slippers) and among the different poses. Barefoot performances produced more stable poses with significantly higher plantar contact areas, smaller COP oscillation areas and smaller anterior-posterior COP oscillations. COP oscillation areas, anterior-posterior COP oscillations and medio-lateral COP velocities indicated that attitude a la second is the least challenging and attitude derriére the most challenging pose. Copyright © 2012 Elsevier B.V. All rights reserved.

  9. Fractal dimension approach in postural control of subjects with Prader-Willi Syndrome

    PubMed Central

    2011-01-01

    Background Static posturography is user-friendly technique suitable for the study of the centre of pressure (CoP) trajectory. However, the utility of static posturography in clinical practice is somehow limited and there is a need for reliable approaches to extract physiologically meaningful information from stabilograms. The aim of this study was to quantify the postural strategy of Prader-Willi patients with the fractal dimension technique in addition to the CoP trajectory analysis in time and frequency domain. Methods 11 adult patients affected by Prader-Willi Syndrome (PWS) and 20 age-matched individuals (Control group: CG) were included in this study. Postural acquisitions were conducted by means of a force platform and the participants were required to stand barefoot on the platform with eyes open and heels at standardized distance and position for 30 seconds. Platform data were analysed in time and frequency domain. Fractal Dimension (FD) was also computed. Results The analysis of CoP vs. time showed that in PWS participants all the parameters were statistically different from CG, with greater displacements along both the antero-posterior and medio-lateral direction and longer CoP tracks. As for frequency analysis, our data showed no significant differences between PWS and CG. FD evidenced that PWS individuals were characterized by greater value in comparison with CG. Conclusions Our data showed that while the analysis in the frequency domain did not seem to explain the postural deficit in PWS, the FD method appears to provide a more informative description of it and to complement and integrate the time domain analysis. PMID:21854639

  10. Insertional Mutations in the Hydrogenase vhc and frc Operons Encoding Selenium-Free Hydrogenases in Methanococcus voltae

    PubMed Central

    Berghofer, Y.; Klein, A.

    1995-01-01

    Methanococcus voltae, which contains four different gene groups that encode [NiFe]-hydrogenases, was transformed with integration vectors to achieve polar inactivation of two of the four hydrogenase operons that encode the selenium-free enzymes Vhc and Frc. Transformants which were selected by their acquired puromycin resistance showed site-specific insertions in either the vhc or frc operon by single crossover events. Southern hybridization revealed tandem integrations of whole vectors in the vhc operon, whereas only one vector copy was found in the frc operon. Northern (RNA) hybridizations showed a pac transcript of defined size, indicating strong termination in front of the hydrogenase genes downstream. In spite of the apparent abolition of expression of selenium-free hydrogenases through these polar insertions, they were not lethal to cells upon growth in selenium-deprived minimal medium, which we had previously shown to strongly induce transcription of the respective operons in M. voltae. Instead, like wild-type control cultures, transformants responded to selenium deprivation only with a reduction in growth rate. We conclude that loss of the potential to express a selenium-free hydrogenase can nevertheless be balanced by very small amounts of selenium hydrogenases under laboratory conditions in which the hydrogen supply is not likely to be a limiting growth factor. PMID:16535019

  11. The chemotaxis regulator pilG of Xylella fastidiosa is required for virulence in Vitis vinifera grapevines

    USDA-ARS?s Scientific Manuscript database

    Type IV pili of X. fastidiosa are regulated by pilG, a response regulator protein putatively involved in chemotaxis-like operon sensing stimuli through signal transduction pathways. To elucidate roles of pilG in pathogenicity of X. fastidiosa, the pilG-deletion mutant and complementary strain contai...

  12. Pantothenic acid biosynthesis in zymomonas

    DOEpatents

    Tao, Luan; Tomb, Jean-Francois; Viitanen, Paul V.

    2014-07-01

    Zymomonas is unable to synthesize pantothenic acid and requires this essential vitamin in growth medium. Zymomonas strains transformed with an operon for expression of 2-dehydropantoate reductase and aspartate 1-decarboxylase were able to grow in medium lacking pantothenic acid. These strains may be used for ethanol production without pantothenic acid supplementation in seed culture and fermentation media.

  13. Characterization of the regulation of a plant polysaccharide utilization operon and its role in biofilm formation in Bacillus subtilis

    PubMed Central

    Habib, Cameron; Yu, Yiyang; Gozzi, Kevin; Ching, Carly; Shemesh, Moshe

    2017-01-01

    The soil bacterium Bacillus subtilis is often found in association with plants in the rhizosphere. Previously, plant polysaccharides have been shown to stimulate formation of root-associated multicellular communities, or biofilms, in this bacterium, yet the underlying mechanism is not fully understood. A five-gene gan operon (ganSPQAB) in B. subtilis has recently been shown to be involved in utilization of the plant-derived polysaccharide galactan. Despite these findings, molecular details about the regulation of the operon and the role of the operon in biofilm formation remain elusive. In this study, we performed comprehensive genetic analyses on the regulation of the gan operon. We show that this operon is regulated both by a LacI-like transcription repressor (GanR), which directly binds to pairs of inverted DNA repeats in the promoter region of the operon, and by the catabolite control protein A (CcpA). Derepression can be triggered by the presence of the inducer β-1,4-galactobiose, a hydrolysis product of galactan, or in situ when B. subtilis cells are associated with plant roots. In addition to the transcriptional regulation, the encoded ß-galactosidase GanA (by ganA), which hydrolyzes ß-1,4-galactobiose into galactose, is inhibited at the enzymatic level by the catalytic product galactose. Thus, the galactan utilization pathway is under complex regulation involving both positive and negative feedback mechanisms in B. subtilis. We discuss about the biological significance of such complex regulation as well as a hypothesis of biofilm induction by galactan via multiple mechanisms. PMID:28617843

  14. Isoprenoid Pyrophosphate-Dependent Transcriptional Regulation of Carotenogenesis in Corynebacterium glutamicum

    PubMed Central

    Henke, Nadja A.; Heider, Sabine A. E.; Hannibal, Silvin; Wendisch, Volker F.; Peters-Wendisch, Petra

    2017-01-01

    Corynebacterium glutamicum is a natural producer of the C50 carotenoid decaprenoxanthin. The crtEcg0722crtBIYEb operon comprises most of its genes for terpenoid biosynthesis. The MarR-type regulator encoded upstream and in divergent orientation of the carotenoid biosynthesis operon has not yet been characterized. This regulator, named CrtR in this study, is encoded in many actinobacterial genomes co-occurring with terpenoid biosynthesis genes. CrtR was shown to repress the crt operon of C. glutamicum since DNA microarray experiments revealed that transcript levels of crt operon genes were increased 10 to 70-fold in its absence. Transcriptional fusions of a promoter-less gfp gene with the crt operon and crtR promoters confirmed that CrtR represses its own gene and the crt operon. Gel mobility shift assays with purified His-tagged CrtR showed that CrtR binds to a region overlapping with the −10 and −35 promoter sequences of the crt operon. Isoprenoid pyrophosphates interfered with binding of CrtR to its target DNA, a so far unknown mechanism for regulation of carotenogenesis. The molecular details of protein-ligand interactions remain to be studied. Decaprenoxanthin synthesis by C. glutamicum wild type was enhanced 10 to 30-fold upon deletion of crtR and was decreased 5 to 6-fold as result of crtR overexpression. Moreover, deletion of crtR was shown as metabolic engineering strategy to improve production of native and non-native carotenoids including lycopene, β-carotene, C.p. 450 and sarcinaxanthin. PMID:28484430

  15. Relatively slow stochastic gene-state switching in the presence of positive feedback significantly broadens the region of bimodality through stabilizing the uninduced phenotypic state.

    PubMed

    Ge, Hao; Wu, Pingping; Qian, Hong; Xie, Xiaoliang Sunney

    2018-03-01

    Within an isogenic population, even in the same extracellular environment, individual cells can exhibit various phenotypic states. The exact role of stochastic gene-state switching regulating the transition among these phenotypic states in a single cell is not fully understood, especially in the presence of positive feedback. Recent high-precision single-cell measurements showed that, at least in bacteria, switching in gene states is slow relative to the typical rates of active transcription and translation. Hence using the lac operon as an archetype, in such a region of operon-state switching, we present a fluctuating-rate model for this classical gene regulation module, incorporating the more realistic operon-state switching mechanism that was recently elucidated. We found that the positive feedback mechanism induces bistability (referred to as deterministic bistability), and that the parameter range for its occurrence is significantly broadened by stochastic operon-state switching. We further show that in the absence of positive feedback, operon-state switching must be extremely slow to trigger bistability by itself. However, in the presence of positive feedback, which stabilizes the induced state, the relatively slow operon-state switching kinetics within the physiological region are sufficient to stabilize the uninduced state, together generating a broadened parameter region of bistability (referred to as stochastic bistability). We illustrate the opposite phenotype-transition rate dependence upon the operon-state switching rates in the two types of bistability, with the aid of a recently proposed rate formula for fluctuating-rate models. The rate formula also predicts a maximal transition rate in the intermediate region of operon-state switching, which is validated by numerical simulations in our model. Overall, our findings suggest a biological function of transcriptional "variations" among genetically identical cells, for the emergence of bistability and transition between phenotypic states.

  16. Prediction of operon-like gene clusters in the Arabidopsis thaliana genome based on co-expression analysis of neighboring genes.

    PubMed

    Wada, Masayoshi; Takahashi, Hiroki; Altaf-Ul-Amin, Md; Nakamura, Kensuke; Hirai, Masami Y; Ohta, Daisaku; Kanaya, Shigehiko

    2012-07-15

    Operon-like arrangements of genes occur in eukaryotes ranging from yeasts and filamentous fungi to nematodes, plants, and mammals. In plants, several examples of operon-like gene clusters involved in metabolic pathways have recently been characterized, e.g. the cyclic hydroxamic acid pathways in maize, the avenacin biosynthesis gene clusters in oat, the thalianol pathway in Arabidopsis thaliana, and the diterpenoid momilactone cluster in rice. Such operon-like gene clusters are defined by their co-regulation or neighboring positions within immediate vicinity of chromosomal regions. A comprehensive analysis of the expression of neighboring genes therefore accounts a crucial step to reveal the complete set of operon-like gene clusters within a genome. Genome-wide prediction of operon-like gene clusters should contribute to functional annotation efforts and provide novel insight into evolutionary aspects acquiring certain biological functions as well. We predicted co-expressed gene clusters by comparing the Pearson correlation coefficient of neighboring genes and randomly selected gene pairs, based on a statistical method that takes false discovery rate (FDR) into consideration for 1469 microarray gene expression datasets of A. thaliana. We estimated that A. thaliana contains 100 operon-like gene clusters in total. We predicted 34 statistically significant gene clusters consisting of 3 to 22 genes each, based on a stringent FDR threshold of 0.1. Functional relationships among genes in individual clusters were estimated by sequence similarity and functional annotation of genes. Duplicated gene pairs (determined based on BLAST with a cutoff of E<10(-5)) are included in 27 clusters. Five clusters are associated with metabolism, containing P450 genes restricted to the Brassica family and predicted to be involved in secondary metabolism. Operon-like clusters tend to include genes encoding bio-machinery associated with ribosomes, the ubiquitin/proteasome system, secondary metabolic pathways, lipid and fatty-acid metabolism, and the lipid transfer system. Copyright © 2012 Elsevier B.V. All rights reserved.

  17. Gait initiation and termination strategies in patients with Prader-Willi syndrome.

    PubMed

    Cimolin, Veronica; Cau, Nicola; Galli, Manuela; Santovito, Cristina; Grugni, Graziano; Capodaglio, Paolo

    2017-05-23

    Gait Initiation (GI) is a functional task representing one of the first voluntary destabilizing behaviours observed in the development of a locomotor pattern as the whole body centre of mass transitions from a large to a small base of support. Conversely, Gait Termination (GT) consists in the transition from walking to standing which, in everyday life, is a very common movement. Compared to normal walking, it requires higher control of postural stability. For a safe GT, the forward movement of the body has to be slowed down to achieve a stable upright position. Stability requirements have to be fulfilled for safe GT. In individuals with Prader-Willi syndrome (PWS), excessive body weight negatively affects the movement, such as walking and posture, but there are no experimental studies about GI and GT in these individuals. The aim of this study was to quantitatively characterise the strategy of patients with PWS during GI and GT using parameters obtained by the Center of Pressure (CoP) track. Twelve patients with PWS, 20 obese (OG) and 19 healthy individuals (HG) were tested using a force platform during the GI and GT tasks. CoP plots were divided into different phases, and duration, length and velocity of the CoP trace in these phases were calculated and compared for each task. As for GI, the results showed a significant reduction of the task duration and lower velocity and CoP length parameters in PWS, compared to OG and HG. In PWS, those parameters were reduced to a higher degree with respect to the OG. During GT, longer durations, similar to OG, were observed in PWS than HG. Velocity is reduced when compared to OG and HG, especially in medio-lateral direction and in the terminal part of GT. From these data, GI appears to be a demanding task in most of its sub-phases for PWS individuals, while GT seems to require caution only towards the end of the task. Breaking the cycle of gait into the phases of GI and GT and implementing specific exercises focusing on weight transfer and foot clearance during the transition phase from the steady condition to gait will possibly improve the effectiveness of rehabilitation and fall and injury prevention.

  18. A new SPE/GC-fid method for the determination of cholesterol oxidation products. Application to subcutaneous fat from Iberian dry-cured ham.

    PubMed

    Narváez-Rivas, Mónica; Pham, Alessandra J; Schilling, M Wes; León-Camacho, Manuel

    2014-05-01

    A new method for the isolation and analysis of cholesterol oxidation products (COPs) using solid phase extraction (SPE) and silica columns was developed using gas chromatography-flame ion detection (GC-FID). The method comprises of saponification and liquid-liquid extraction of the unsaponifiable fraction prior to the isolation and derivatization of the COPs to trimethylsilyl ethers. The COPs used in this study are cholestane-5α-6α-epoxide, cholestane-3β-5α-6β-triol, 25-hydroxycholesterol and 5-cholesten-3β-ol-7-one. In order to identify the COPs fraction a GC-ion-trap-mass spectrometry experiment were conducted using authentic standards to verify the presence of the COPs. The method was effective at rapidly separating the COPs (25 min run). Calibration curves were linear with the LODs and LOQs bellow 0.03 and 0.07 mgkg(-1) for all cases, respectively. This methodology gave a total recovery for every compound that was used in the study. Betulin was used as an internal standard to monitor the recovery. The method was validated with a standard mixture of COPs. The method has been applied to characterize the COP fraction of subcutaneous fat from Iberian dry-cured ham. Cholestane-5α-6α-epoxide, cholestane-3β-5α-6β-triol, 25-hydroxycholesterol and 5-cholesten-3β-ol-7-one have been identified for the first time in these samples. Copyright © 2014 Elsevier B.V. All rights reserved.

  19. Sixteen-Day Bedrest Significantly Increases Plasma Colloid Osmotic Pressure

    NASA Technical Reports Server (NTRS)

    Hargens, Alan R.; Hsieh, S. T.; Murthy, G.; Ballard, R. E.; Convertino, V. A.; Wade, Charles E. (Technical Monitor)

    1994-01-01

    Upon exposure to microgravity, astronauts lose up to 10% of their total plasma volume, which may contribute to orthostatic intolerance after space flight. Because plasma colloid osmotic pressure (COP) is a primary factor maintaining plasma volume, our objective was to measure time course changes in COP during microgravity simulated by 6 deg. head-down tilt (HDT). Seven healthy male subjects (30-55 years of age) were placed in HDT for 16 days. For the purpose of another study, three of the seven subjects were chosen to exercise on a cycle ergometer on day 16. Blood samples were drawn immediately before bedrest on day 14 of bedrest, 18-24 hours following exercise while all subjects were still in HDT and 1 hour following bedrest termination. Plasma COP was measured in all 20 microliter EDTA-treated samples using an osmometer fitted with a PM 30 membrane. Data were analyzed with paired and unpaired t-tests. Plasma COP on day 14 of bedrest (29.9 +/- 0.69 mmHg) was significantly higher (p less than 0.005) than the control, pre-bedrest value (23.1 +/- 0.76 mmHg). At one hour of upright recovery after HDT, plasma COP remained significantly elevated (exercise: 26.9 +/- 0.87 mmHg; no exercise: 26.3 +/- 0.85 mmHg). Additionally, exercise had no significant effect on plasma COP 18-24 hours following exercise (exercise: 27.8 +/- 1.09 mmHg; no exercise: 27.1 +/- 0.78 mmHg). Our results demonstrate that plasma COP increases significantly with microgravity simulated by HDT. However, preliminary results indicate exercise during HDT does not significantly affect plasma COP.

  20. Relationships between the center of pressure and the movements of the ankle and knee joints during the stance phase in patients with severe medial knee osteoarthritis.

    PubMed

    Fukaya, Takashi; Mutsuzaki, Hirotaka; Okubo, Tomoyuki; Mori, Koichi; Wadano, Yasuyoshi

    2016-08-01

    The knee joint movement during the stance phase is affected by altered ankle movement and the center of pressure (COP). However the relationships between changes in the center of pressure (COP) and the altered kinematics and kinetics of the ankle and knee joints in patients with osteoarthritis (OA) of the knee are not well understood. The purpose of this study was to determine the relationships between changes in the COP and the altered kinematic and kinetic variables in ankle and knee joints during the stance phase in patients with medial knee OA. Fourteen patients with knee OA (21 knees) and healthy subjects were assessed by gait analysis using an eight-camera motion analysis system to record forward and lateral shifts in the COP and the angle and net internal moments of the knee and ankle joint. Spearman rank-correlation coefficients were used to determine the relationship between these results. In knees with medial OA, lateral shifts in the COP were correlated with knee flexion angle. Lateral shifts in the COP were correlated with the second peak of the knee extensor moment and correlated with the knee abductor moment. In patients with medial knee OA, lateral shifts in the COP were negatively correlated with the kinematic and kinetic variables in the sagittal plane of the knee joints. Controlling such lateral shifts in the COP may thus be an effective intervention for mechanical loads on the knee during the stance phase in patients with knee OA. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. β-COP as a Component of Transport Vesicles for HDL Apolipoprotein-Mediated Cholesterol Exocytosis

    PubMed Central

    Ma, Weilie; Lin, Margarita; Ding, Hang; Lin, Guorong; Zhang, Zhizhen

    2016-01-01

    Objective HDL and its apolipoproteins protect against atherosclerotic disease partly by removing excess cholesterol from macrophage foam cells. But the underlying mechanisms of cholesterol clearance are still not well defined. We investigated roles of vesicle trafficking of coatomer β-COP in delivering cholesterol to the cell surface during apoA-1 and apoE-mediated lipid efflux from fibroblasts and THP-1 macrophages. Methods shRNA knockout, confocal and electron microscopy and biochemical analysis were used to investigate the roles of β-COP in apolipoprotein-mediated cholesterol efflux in fibroblasts and THP-1 macrophages. Results We showed that β-COP knockdown by lentiviral shRNA resulted in reduced apoA-1-mediated cholesterol efflux, while increased cholesterol accumulation and formation of larger vesicles were observed in THP-1 macrophages by laser scanning confocal microscopy. Immunogold electron microscopy showed that β-COP appeared on the membrane protrusion complexes and colocalized with apoA-1 or apoE during cholesterol efflux. This was associated with releasing heterogeneous sizes of small particles into the culture media of THP-1 macrophage. Western blotting also showed that apoA-1 promotes β-COP translocation to the cell membrane and secretion into culture media, in which a total of 17 proteins were identified by proteomics. Moreover, β-COP exclusively associated with human plasma HDL fractions. Conclusion ApoA-1 and apoE promoted transport vesicles consisting of β-COP and other candidate proteins to exocytose cholesterol, forming the protrusion complexes on cell surface, which were then released from the cell membrane as small particles to media. PMID:26986486

  2. Salmonella enterica Serovar Enteritidis Core O Polysaccharide Conjugated to H:g,m Flagellin as a Candidate Vaccine for Protection against Invasive Infection with S. Enteritidis▿†

    PubMed Central

    Simon, Raphael; Tennant, Sharon M.; Wang, Jin Y.; Schmidlein, Patrick J.; Lees, Andrew; Ernst, Robert K.; Pasetti, Marcela F.; Galen, James E.; Levine, Myron M.

    2011-01-01

    Nontyphoidal Salmonella enterica serovars Enteritidis and Typhimurium are a common cause of gastroenteritis but also cause invasive infections and enteric fever in certain hosts (young children in sub-Saharan Africa, the elderly, and immunocompromised individuals). Salmonella O polysaccharides (OPS) and flagellar proteins are virulence factors and protective antigens. The surface polysaccharides of Salmonella are poorly immunogenic and do not confer immunologic memory, limitations overcome by covalently attaching them to carrier proteins. We conjugated core polysaccharide-OPS (COPS) of Salmonella Enteritidis lipopolysaccharide (LPS) to flagellin protein from the homologous strain. COPS and flagellin were purified from a genetically attenuated (ΔguaBA) “reagent strain” (derived from an isolate from a patient with clinical bacteremia) engineered for increased flagellin production (ΔclpPX). Conjugates were constructed by linking flagellin monomers or polymers at random COPS hydroxyls with various polysaccharide/protein ratios by 1-cyano-4-dimethylaminopyridinium tetrafluoroborate (CDAP) or at the 3-deoxy-d-manno-octulosonic acid (KDO) terminus by thioether chemistry. Mice immunized on days 0, 28, and 56 with COPS-flagellin conjugates mounted higher anti-LPS IgG levels than mice receiving unconjugated COPS and exhibited high antiflagellin IgG; anti-LPS and antiflagellin IgG levels increased following booster doses. Antibodies generated by COPS-flagellin conjugates mediated opsonophagocytosis of S. Enteritidis cells into mouse macrophages. Mice immunized with flagellin alone, COPS-CRM197, or COPS-flagellin conjugates were significantly protected from lethal challenge with wild-type S. Enteritidis (80 to 100% vaccine efficacy). PMID:21807909

  3. Copolymer-1 enhances cognitive performance in young adult rats

    PubMed Central

    Meneses, Alfredo; Cruz-Martínez, Yolanda; Anaya-Jiménez, Rosa María; Liy-Salmerón, Gustavo; Carvajal, Horacio Guillermo; Ponce-López, Maria Teresa

    2018-01-01

    Cognitive impairment is a dysfunction observed as a sequel of various neurodegenerative diseases, as well as a concomitant element in the elderly stages of life. In clinical settings, this malfunction is identified as mild cognitive impairment. Previous studies have suggested that cognitive impairment could be the result of a reduction in the expression of brain-derived neurotrophic factor (BDNF) and/or immune dysfunction. Copolymer-1 (Cop-1) is an FDA-approved synthetic peptide capable of inducing the activation of Th2/3 cells, which are able to release BDNF, as well as to migrate and accumulate in the brain. In this study, we evaluated the effect of Cop-1 immunization on improvement of cognition in adult rats. For this purpose, we performed four experiments. We evaluated the effect of Cop-1 immunization on learning/memory using the Morris water maze for spatial memory and autoshaping for associative memory in 3- or 6-month-old rats. BDNF concentrations at the hippocampus were determined by ELISA. Cop-1 immunization induced a significant improvement of spatial memory and associative memory in 6-month-old rats. Likewise, Cop-1 improved spatial memory and associative memory when animals were immunized at 3 months and evaluated at 6 months old. Additionally, Cop-1 induced a significant increase in BDNF levels at the hippocampus. To our knowledge, the present investigation reports the first instance of Cop-1 treatment enhancing cognitive function in normal young adult rats, suggesting that Cop-1 may be a practical therapeutic strategy potentially useful for age- or disease-related cognitive impairment. PMID:29494605

  4. Flavobacterium johnsoniae sprB Is Part of an Operon Spanning the Additional Gliding Motility Genes sprC, sprD, and sprF ▿ †

    PubMed Central

    Rhodes, Ryan G.; Nelson, Shawn S.; Pochiraju, Soumya; McBride, Mark J.

    2011-01-01

    Cells of Flavobacterium johnsoniae move rapidly over surfaces by a process known as gliding motility. Gld proteins are thought to comprise the gliding motor that propels cell surface adhesins, such as the 669-kDa SprB. A novel protein secretion apparatus called the Por secretion system (PorSS) is required for assembly of SprB on the cell surface. Genetic and molecular analyses revealed that sprB is part of a seven-gene operon spanning 29.3 kbp of DNA. In addition to sprB, three other genes of this operon (sprC, sprD, and sprF) are involved in gliding. Mutations in sprB, sprC, sprD, and sprF resulted in cells that failed to form spreading colonies on agar but that exhibited some motility on glass in wet mounts. SprF exhibits some similarity to Porphyromonas gingivalis PorP, which is required for secretion of gingipain protease virulence factors via the P. gingivalis PorSS. F. johnsoniae sprF mutants produced SprB protein but were defective in localization of SprB to the cell surface, suggesting a role for SprF in secretion of SprB. The F. johnsoniae PorSS is involved in secretion of extracellular chitinase in addition to its role in secretion of SprB. SprF was not needed for chitinase secretion and may be specifically required for SprB secretion by the PorSS. Cells with nonpolar mutations in sprC or sprD produced and secreted SprB and propelled it rapidly along the cell surface. Multiple paralogs of sprB, sprC, sprD, and sprF are present in the genome, which may explain why mutations in sprB, sprC, sprD, and sprF do not result in complete loss of motility and suggests the possibility that semiredundant SprB-like adhesins may allow movement of cells over different surfaces. PMID:21131497

  5. 78 FR 64979 - Agency Information Collection Activities; Revision of a Previously Approved Collection, with...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-10-30

    ... Collection, with Change; Comments Requested: COPS Progress Report ACTION: 60-Day Notice. The Department of Justice (DOJ) Office of Community Oriented Policing Services (COPS) will be submitting the following...; comments requested. (2) Title of the Form/Collection: COPS Progress Report. (3) Agency form number, if any...

  6. 76 FR 10064 - Agency Information Collection Activities: Extension of a Previously Approved Collection; Comments...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-02-23

    ... Collection Under Review: COPS Progress Report. The Department of Justice (DOJ) Office of Community Oriented Policing Services (COPS) will be submitting the following information collection request to the Office of... Form/Collection: COPS Progress Report. (3) Agency form number, if any, and the applicable component of...

  7. CoPs Facing Rationalization: The Politics of Community Reproduction

    ERIC Educational Resources Information Center

    Kilskar, Stine Skaufel; Ingvaldsen, Jonas A.; Valle, Nina

    2018-01-01

    Purpose: This paper aims to explore the relationship between the contemporary forms of manufacturing rationalization and the reproduction of communities of practice (CoPs) centred on tasks and craft. Building on critical literature highlighting the tensions between CoPs and rationalization, this paper aims to develop a nuanced account of how CoPs…

  8. School-Based Partnerships: A Problem-Solving Strategy. COPS Innovations

    ERIC Educational Resources Information Center

    Uchida, Craig D.; Solomon, Shellie; Katz, Charles M.; Pappas, Cynthia E.

    2006-01-01

    The Community Oriented Policing Services (COPS) Office funded the School-Based Partnerships (SBP) grant program for the purpose of partnering law enforcement agencies with schools to address crime and disorder problems in and around middle and high schools. In Fiscal Years 1998 and 1999, the COPS Office awarded 275 law enforcement agencies more…

  9. Modified combination of platelet count and neutrophil "to" lymphocyte ratio as a prognostic factor in patients with advanced head and neck cancer.

    PubMed

    Nakayama, Masahiro; Gosho, Masahiko; Hirose, Yuki; Nishimura, Bungo; Tanaka, Shuho; Tabuchi, Keiji; Okubo, Hideki; Wada, Tetsuro; Hara, Akira

    2018-06-01

    We evaluated the prognostic potential of the combination of platelet count and neutrophil to lymphocyte ratio (COP-NLR) in patients with advanced head and neck cancer. We proposed a modified COP-NLR scoring system defined as follows: score 0 (platelet count level <300 × 10 9 /L and NLR <3); score 1 (platelet count level ≥300 × 10 9 /L and NLR <3); and score 2 (NLR ≥3). We assessed whether the modified scoring system had better performance as an indicator of prognosis than the existing COP-NLR scoring system (original and 4-group scores). A total of 248 patients were enrolled. The Akaike Information Criterion value with the modified COP-NLR score was the smallest among the 3 models. The 3-year survival rates according to the modified COP-NLR scores of 0, 1, and 2 were 80.6%, 59.9%, and 23.8%, respectively. The modified COP-NLR score is a useful prognostic marker in patients with advanced head and neck cancer. © 2018 Wiley Periodicals, Inc.

  10. The complete nucleotide sequence of the glnALG operon of Escherichia coli K12.

    PubMed Central

    Miranda-Ríos, J; Sánchez-Pescador, R; Urdea, M; Covarrubias, A A

    1987-01-01

    The nucleotide sequence of the E. coli glnALG operon has been determined. The glnL (ntrB) and glnG (ntrC) genes present a high homology, at the nucleotide and aminoacid levels, with the corresponding genes of Klebsiella pneumoniae. The predicted aminoacid sequence for glutamine synthetase allowed us to locate some of the enzyme domains. The structure of this operon is discussed. PMID:2882477

  11. Fast vaccine design and development based on correlates of protection (COPs)

    PubMed Central

    van Els, Cécile; Mjaaland, Siri; Næss, Lisbeth; Sarkadi, Julia; Gonczol, Eva; Smith Korsholm, Karen; Hansen, Jon; de Jonge, Jørgen; Kersten, Gideon; Warner, Jennifer; Semper, Amanda; Kruiswijk, Corine; Oftung, Fredrik

    2014-01-01

    New and reemerging infectious diseases call for innovative and efficient control strategies of which fast vaccine design and development represent an important element. In emergency situations, when time is limited, identification and use of correlates of protection (COPs) may play a key role as a strategic tool for accelerated vaccine design, testing, and licensure. We propose that general rules for COP-based vaccine design can be extracted from the existing knowledge of protective immune responses against a large spectrum of relevant viral and bacterial pathogens. Herein, we focus on the applicability of this approach by reviewing the established and up-coming COPs for influenza in the context of traditional and a wide array of new vaccine concepts. The lessons learnt from this field may be applied more generally to COP-based accelerated vaccine design for emerging infections. PMID:25424803

  12. Development of a glycoconjugate vaccine to prevent invasive Salmonella Typhimurium infections in sub-Saharan Africa

    PubMed Central

    Baliban, Scott M.; Yang, Mingjun; Ramachandran, Girish; Curtis, Brittany; Shridhar, Surekha; Laufer, Rachel S.; Wang, Jin Y.; Van Druff, John; Higginson, Ellen E.; Hegerle, Nicolas; Varney, Kristen M.; Galen, James E.; Tennant, Sharon M.; Lees, Andrew; MacKerell, Alexander D.; Levine, Myron M.; Simon, Raphael

    2017-01-01

    Invasive infections associated with non-typhoidal Salmonella (NTS) serovars Enteritidis (SE), Typhimurium (STm) and monophasic variant 1,4,[5],12:i:- are a major health problem in infants and young children in sub-Saharan Africa, and currently, there are no approved human NTS vaccines. NTS O-polysaccharides and flagellin proteins are protective antigens in animal models of invasive NTS infection. Conjugates of SE core and O-polysaccharide (COPS) chemically linked to SE flagellin have enhanced the anti-COPS immune response and protected mice against fatal challenge with a Malian SE blood isolate. We report herein the development of a STm glycoconjugate vaccine comprised of STm COPS conjugated to the homologous serovar phase 1 flagellin protein (FliC) with assessment of the role of COPS O-acetyls for functional immunity. Sun-type COPS conjugates linked through the polysaccharide reducing end to FliC were more immunogenic and protective in mice challenged with a Malian STm blood isolate than multipoint lattice conjugates (>95% vaccine efficacy [VE] versus 30–43% VE). Immunization with de-O-acetylated STm-COPS conjugated to CRM197 provided significant but reduced protection against STm challenge compared to mice immunized with native STm-COPS:CRM197 (63–74% VE versus 100% VE). Although OPS O-acetyls were highly immunogenic, post-vaccination sera that contained various O-acetyl epitope-specific antibody profiles displayed similar in vitro bactericidal activity when equivalent titers of anti-COPS IgG were assayed. In-silico molecular modeling further indicated that STm OPS forms a single dominant conformation, irrespective of O-acetylation, in which O-acetyls extend outward and are highly solvent exposed. These preclinical results establish important quality attributes for an STm vaccine that could be co-formulated with an SE-COPS:FliC glycoconjugate as a bivalent NTS vaccine for use in sub-Saharan Africa. PMID:28388624

  13. Exploratory behavior during stance persists with visual feedback.

    PubMed

    Murnaghan, C D; Horslen, B C; Inglis, J T; Carpenter, M G

    2011-11-10

    Recent evidence showing center of pressure (COP) displacements increase following an external stabilization of the center of mass (COM) supports the theory that postural sway may be exploratory and serve as a means of acquiring sensory information. The aim of the current study was to further test this theory and rule out potential confounding effects of sensory illusions or motor drift on prior observations. Participants stood as still as possible in an apparatus which allowed movements of the COM to be stabilized ("locked") without subject awareness, and they were provided real-time visual feedback of their COM or COP throughout the trial. If there was an influence of sensory illusions or motor drift, we hypothesized that the change in COP displacement with locking would be reduced when participants were provided visual confirmation of COM stabilization (COM feedback), or when they were aware of the position of the COP throughout the trial (COP feedback). Confirming our previous results, increases in COP displacement were observed when movements of the COM were stabilized. In addition, our results showed that increases in COP displacement could not be explained by the presence of sensory illusions or motor drift, since increases in COP were observed despite being provided convincing evidence that the COM had been stabilized, and when participants were aware of their COP position throughout the trial. These results provide further support for an exploratory role of postural sway. The theoretical basis of current clinical practices designed to deal with balance control deficits due to age or disease is typically based on the opinion that increases in sway are a consequence of a failing balance control system. Our results suggest that this may not be the case, and if sway is in fact exploratory, a serious re-evaluation of current clinical practices may be warranted. Copyright © 2011 IBRO. Published by Elsevier Ltd. All rights reserved.

  14. An explorative investigation of functional differences in plantar center of pressure of four foot types using sample entropy method.

    PubMed

    Mei, Zhanyong; Ivanov, Kamen; Zhao, Guoru; Li, Huihui; Wang, Lei

    2017-04-01

    In the study of biomechanics of different foot types, temporal or spatial parameters derived from plantar pressure are often used. However, there is no comparative study of complexity and regularity of the center of pressure (CoP) during the stance phase among pes valgus, pes cavus, hallux valgus and normal foot. We aim to analyze whether CoP sample entropy characteristics differ among these four foot types. In our experiment participated 40 subjects with normal feet, 40 with pes cavus, 19 with pes valgus and 36 with hallux valgus. A Footscan ® system was used to collect CoP data. We used sample entropy to quantify several parameters of the investigated four foot types. These are the displacement in medial-lateral (M/L) and anterior-posterior (A/P) directions, as well as the vertical ground reaction force of CoP during the stance phase. To fully examine the potential of the sample entropy method for quantification of CoP components, we provide results for two cases: calculating the sample entropy of normalized CoP components, as well as calculating it using the raw data of CoP components. We also explored what are the optimal values of parameters m (the matching length) and r (the tolerance range) when calculating the sample entropy of CoP data obtained during the stance phases. According to statistical results, some factors significantly influenced the sample entropy of CoP components. The sample entropies of non-normalized A/P values for the left foot, as well as for the right foot, were different between the normal foot and pes valgus, and between the normal foot and hallux valgus. The sample entropy of normalized M/L displacement of the right foot was different between the normal foot and pes cavus. The measured variable for A/P and M/L displacements could serve for the study of foot function.

  15. Footwear and Foam Surface Alter Gait Initiation of Typical Subjects

    PubMed Central

    Vieira, Marcus Fraga; Sacco, Isabel de Camargo Neves; Nora, Fernanda Grazielle da Silva Azevedo; Rosenbaum, Dieter; Lobo da Costa, Paula Hentschel

    2015-01-01

    Gait initiation is the task commonly used to investigate the anticipatory postural adjustments necessary to begin a new gait cycle from the standing position. In this study, we analyzed whether and how foot-floor interface characteristics influence the gait initiation process. For this purpose, 25 undergraduate students were evaluated while performing a gait initiation task in three experimental conditions: barefoot on a hard surface (barefoot condition), barefoot on a soft surface (foam condition), and shod on a hard surface (shod condition). Two force plates were used to acquire ground reaction forces and moments for each foot separately. A statistical parametric mapping (SPM) analysis was performed in COP time series. We compared the anterior-posterior (AP) and medial-lateral (ML) resultant center of pressure (COP) paths and average velocities, the force peaks under the right and left foot, and the COP integral x force impulse for three different phases: the anticipatory postural adjustment (APA) phase (Phase 1), the swing-foot unloading phase (Phase 2), and the support-foot unloading phase (Phase 3). In Phase 1, significantly smaller ML COP paths and velocities were found for the shod condition compared to the barefoot and foam conditions. Significantly smaller ML COP paths were also found in Phase 2 for the shod condition compared to the barefoot and foam conditions. In Phase 3, increased AP COP velocities were found for the shod condition compared to the barefoot and foam conditions. SPM analysis revealed significant differences for vector COP time series in the shod condition compared to the barefoot and foam conditions. The foam condition limited the impulse-generating capacity of COP shift and produced smaller ML force peaks, resulting in limitations to body-weight transfer from the swing to the support foot. The results suggest that footwear and a soft surface affect COP and impose certain features of gait initiation, especially in the ML direction of Phase 1. PMID:26270323

  16. A castor oil-containing dental luting agent: effects of cyclic loading and storage time on flexural strength

    PubMed Central

    DERCELI, Juliana dos Reis; FAIS, Laiza Maria Grassi; PINELLI, Lígia Antunes Pereira

    2014-01-01

    Favorable results in the use of castor oil polyurethane (COP) as pulp capping, membrane material, sealer, mouthwash and in bone repair, associated with the fact that Ricinus communis is not derived from petroleum and it is abundant in Brazil, encourage researches in the development of luting agents. Objectives This study compared the flexural strength (FS) of a castor oil-containing dental luting agent with a weight percentage of 10% (wt%) of calcium carbonate (COP10) with RelyX ARC (RX) after mechanical cycling (MC) and distilled water storage. Material and Methods Sixty-four specimens (25x2x2 mm) were fabricated and divided into two groups, COP10 and RX (control). Each group was divided into 4 subgroups (n=8) according to the storage time, 24 hours (24 h) or 60 days (60 d), and the performance (MC+FS) or not (only FS) of the mechanical cycling test. The FS (10 kN; 0.5 mm/min) and MC tests (10,000 cycles, 5 Hz, 0.5 mm/min) were carried out using an MTS-810 machine. The data were analyzed using ANOVA (α=0.05). Results The obtained FS (MPa) values were: COP10 24h- 19.04±2.41; COP10 60d- 17.92±3.54; RX 24h- 75.19±3.43; RX 60d- 88.77±6.89. All the RX specimens submitted to MC fractured, while the values for COP10 after MC were as follows: COP10 24h- 17.90±1.87 and COP10 60d- 18.60±1.60. Conclusions A castor oil-containing dental luting agent with a weight percentage of 10% (wt%) of calcium carbonate is resistant to mechanical cycling without decreases in flexural strength. However, mean COP10 showed only about 25% of the RelyX ARC mean flexural strength. PMID:25591018

  17. Development of a glycoconjugate vaccine to prevent invasive Salmonella Typhimurium infections in sub-Saharan Africa.

    PubMed

    Baliban, Scott M; Yang, Mingjun; Ramachandran, Girish; Curtis, Brittany; Shridhar, Surekha; Laufer, Rachel S; Wang, Jin Y; Van Druff, John; Higginson, Ellen E; Hegerle, Nicolas; Varney, Kristen M; Galen, James E; Tennant, Sharon M; Lees, Andrew; MacKerell, Alexander D; Levine, Myron M; Simon, Raphael

    2017-04-01

    Invasive infections associated with non-typhoidal Salmonella (NTS) serovars Enteritidis (SE), Typhimurium (STm) and monophasic variant 1,4,[5],12:i:- are a major health problem in infants and young children in sub-Saharan Africa, and currently, there are no approved human NTS vaccines. NTS O-polysaccharides and flagellin proteins are protective antigens in animal models of invasive NTS infection. Conjugates of SE core and O-polysaccharide (COPS) chemically linked to SE flagellin have enhanced the anti-COPS immune response and protected mice against fatal challenge with a Malian SE blood isolate. We report herein the development of a STm glycoconjugate vaccine comprised of STm COPS conjugated to the homologous serovar phase 1 flagellin protein (FliC) with assessment of the role of COPS O-acetyls for functional immunity. Sun-type COPS conjugates linked through the polysaccharide reducing end to FliC were more immunogenic and protective in mice challenged with a Malian STm blood isolate than multipoint lattice conjugates (>95% vaccine efficacy [VE] versus 30-43% VE). Immunization with de-O-acetylated STm-COPS conjugated to CRM197 provided significant but reduced protection against STm challenge compared to mice immunized with native STm-COPS:CRM197 (63-74% VE versus 100% VE). Although OPS O-acetyls were highly immunogenic, post-vaccination sera that contained various O-acetyl epitope-specific antibody profiles displayed similar in vitro bactericidal activity when equivalent titers of anti-COPS IgG were assayed. In-silico molecular modeling further indicated that STm OPS forms a single dominant conformation, irrespective of O-acetylation, in which O-acetyls extend outward and are highly solvent exposed. These preclinical results establish important quality attributes for an STm vaccine that could be co-formulated with an SE-COPS:FliC glycoconjugate as a bivalent NTS vaccine for use in sub-Saharan Africa.

  18. The Yersinia pestis caf1M1A1 fimbrial capsule operon promotes transmission by flea bite in a mouse model of bubonic plague.

    PubMed

    Sebbane, Florent; Jarrett, Clayton; Gardner, Donald; Long, Daniel; Hinnebusch, B Joseph

    2009-03-01

    Plague is a zoonosis transmitted by fleas and caused by the gram-negative bacterium Yersinia pestis. During infection, the plasmidic caf1M1A1 operon that encodes the Y. pestis F1 protein capsule is highly expressed, and anti-F1 antibodies are protective. Surprisingly, the capsule is not required for virulence after injection of cultured bacteria, even though it is an antiphagocytic factor and capsule-deficient Y. pestis strains are rarely isolated. We found that a caf-negative Y. pestis mutant was not impaired in either flea colonization or virulence in mice after intradermal inoculation of cultured bacteria. In contrast, absence of the caf operon decreased bubonic plague incidence after a flea bite. Successful development of plague in mice infected by flea bite with the caf-negative mutant required a higher number of infective bites per challenge. In addition, the mutant displayed a highly autoaggregative phenotype in infected liver and spleen. The results suggest that acquisition of the caf locus via horizontal transfer by an ancestral Y. pestis strain increased transmissibility and the potential for epidemic spread. In addition, our data support a model in which atypical caf-negative strains could emerge during climatic conditions that favor a high flea burden. Human infection with such strains would not be diagnosed by the standard clinical tests that detect F1 antibody or antigen, suggesting that more comprehensive surveillance for atypical Y. pestis strains in plague foci may be necessary. The results also highlight the importance of studying Y. pestis pathogenesis in the natural context of arthropod-borne transmission.

  19. The atlA operon of Streptococcus mutans: role in autolysin maturation and cell surface biogenesis.

    PubMed

    Ahn, Sang-Joon; Burne, Robert A

    2006-10-01

    The Smu0630 protein (AtlA) was recently shown to be involved in cell separation, biofilm formation, and autolysis. Here, transcriptional studies revealed that atlA is part of a multigene operon under the control of at least three promoters. The morphology and biofilm-forming capacity of a nonpolar altA mutant could be restored to that of the wild-type strain by adding purified AtlA protein to the medium. A series of truncated derivatives of AtlA revealed that full activity required the C terminus and repeat regions. AtlA was cell associated and readily extractable from with sodium dodecyl sulfate. Of particular interest, the surface protein profile of AtlA-deficient strains was dramatically altered compared to the wild-type strain, as was the nature of the association of the multifunctional adhesin P1 with the cell wall. In addition, AtlA-deficient strains failed to develop competence as effectively as the parental strain. Mutation of thmA, which can be cotranscribed with atlA and encodes a putative pore-forming protein, resulted in a phenotype very similar to that of the AtlA-deficient strain. ThmA was also shown to be required for efficient processing of AtlA to its mature form, and treatment of the thmA mutant strain with full-length AtlA protein did not restore normal cell separation and biofilm formation. The effects of mutating other genes in the operon on cell division, biofilm formation, or AtlA biogenesis were not as profound. This study reveals that AtlA is a surface-associated protein that plays a critical role in the network connecting cell surface biogenesis, biofilm formation, genetic competence, and autolysis.

  20. Introducing a multimedia course to enhance health professionals' skills to facilitate communities of practice.

    PubMed

    Jakubec, Sonya L; Parboosingh, John; Colvin, Barbara

    2014-01-01

    Scholarship about communities of practice (COP) is uncovering evidence that interactivity between community members contributes to improvement in practice. Leadership and facilitation are crucial elements of successful COP implementation. The purpose of this paper is to describe an innovative COP facilitator's course and report on the experiences of participants in the first course. In response to this need and emerging evidence, an on-line COP facilitator's course was developed and implemented in Alberta, Canada, in 2011. This course included a home-based COP practicum, introductory face-to-face session, an on-line discussion board moderated by faculty and on-line learning modules. Evaluation of the course was formalized in a qualitative study incorporating content analysis of postings, semi-structured interviews of successful participants and narrative responses to questions in a post course survey. A total of 15 of 22 participants perceived they acquired basic knowledge about community facilitation by completing the self-learning modules and assignments. Many did not establish home-based COP and only partially participated in the interactive components of the course. Six participants successfully completed the course by establishing home-based COP and actively participating in the social and interactive components of the course. They perceived they met course objectives and greatly benefited from participation in the course, in particular when they pushed themselves to facilitate in new and different ways, and when they were actively engaged with their home-based COP where they could practice and receive feedback. While the main reasons why participants dropped out or failed to complete all course components were reported, the experiences and perceptions of six participants who successfully completed all course components form the major part of the evaluation of the course and hence introduce bias. A more in depth analysis of why learners are reluctant to engage in participatory learning could be the focus of further studies. The following key recommendations emerged in the study alongside recommendations for further study of best practices in supporting COP facilitation. First, a formal interview before enrollment into the COP facilitator's course is recommended to reinforce the comprehensiveness, time commitment and the practical applications intended within the course. Second, methods of "aggressive facilitation" with skilled COP facilitators can best model facilitation to those involved in the course. Third, supporting course participants to trial out a diversity of community facilitation skills in the safety of the course is crucial to success. The collaboration, networking and interactivity of interdisciplinary health care workers is of tremendous consequence to health outcomes and a vital concern to practitioners and administrators. Little is currently understood of the leadership and facilitation of the COP models and these discoveries lend a timely contribution to the field.

  1. Structure, Regulation, and Putative Function of the Arginine Deiminase System of Streptococcus suis

    PubMed Central

    Gruening, Petra; Fulde, Marcus; Valentin-Weigand, Peter; Goethe, Ralph

    2006-01-01

    Streptococcus suis is an important cause of infectious diseases in young pigs. Little is known about the virulence factors or protective antigens of S. suis. Recently, we have identified two proteins of the arginine deiminase system (ADS) of S. suis, which were temperature induced and expressed on the streptococcal surface (N. Winterhoff, R. Goethe, P. Gruening, M. Rohde, H. Kalisz, H. E. Smith, and P. Valentin-Weigand, J. Bacteriol. 184:6768-6776, 2002). In the present study, we analyzed the complete ADS of S. suis. Due to their homologies to the recently published S. gordonii ADS genes, the genes for arginine deiminase, ornithine carbamoyl-transferase, and carbamate kinase, which were previously designated adiS, octS, and ckS, respectively, were renamed arcA, arcB, and arcC, respectively. Our data revealed that arcA, arcB, and arcC of the S. suis ADS are transcribed from an operon (arcABC operon). Additionally, putative ADS-associated genes were cloned and sequenced which, however, did not belong to the arcABC operon. These were the flpS gene upstream of the arcABC operon with homology to the flp transcription regulator of S. gordonii and the arcD, arcT, arcH, and argR genes downstream of the arcABC operon with high homologies to a putative arginine-ornithine antiporter, a putative dipeptidase of S. gordonii, a putative β-N-acetylhexosaminidase of S. pneumoniae, and a putative arginine repressor of S. gordonii, respectively. The transcriptional start point of the arcABC operon was determined, and promoter analysis provided evidence that multiple factors contribute to the regulation of the ADS. Thus, a putative binding site for a transcription regulator of the Crp/Fnr family, an ArgR-binding site, and two cis-acting catabolite response elements were identified in the promoter-operator region of the operon. Consistent with this, we could demonstrate that the ADS of S. suis is inducible by arginine and reduced O2 tension and subject to carbon catabolite repression. Furthermore, comparing an arcA knockout mutant in which expression of the three operon-encoded proteins was abolished with the parental wild-type strain showed that the arcABC operon of S. suis contributes to survival under acidic conditions. PMID:16385025

  2. Structure, regulation, and putative function of the arginine deiminase system of Streptococcus suis.

    PubMed

    Gruening, Petra; Fulde, Marcus; Valentin-Weigand, Peter; Goethe, Ralph

    2006-01-01

    Streptococcus suis is an important cause of infectious diseases in young pigs. Little is known about the virulence factors or protective antigens of S. suis. Recently, we have identified two proteins of the arginine deiminase system (ADS) of S. suis, which were temperature induced and expressed on the streptococcal surface (N. Winterhoff, R. Goethe, P. Gruening, M. Rohde, H. Kalisz, H. E. Smith, and P. Valentin-Weigand, J. Bacteriol. 184:6768-6776, 2002). In the present study, we analyzed the complete ADS of S. suis. Due to their homologies to the recently published S. gordonii ADS genes, the genes for arginine deiminase, ornithine carbamoyl-transferase, and carbamate kinase, which were previously designated adiS, octS, and ckS, respectively, were renamed arcA, arcB, and arcC, respectively. Our data revealed that arcA, arcB, and arcC of the S. suis ADS are transcribed from an operon (arcABC operon). Additionally, putative ADS-associated genes were cloned and sequenced which, however, did not belong to the arcABC operon. These were the flpS gene upstream of the arcABC operon with homology to the flp transcription regulator of S. gordonii and the arcD, arcT, arcH, and argR genes downstream of the arcABC operon with high homologies to a putative arginine-ornithine antiporter, a putative dipeptidase of S. gordonii, a putative beta-N-acetylhexosaminidase of S. pneumoniae, and a putative arginine repressor of S. gordonii, respectively. The transcriptional start point of the arcABC operon was determined, and promoter analysis provided evidence that multiple factors contribute to the regulation of the ADS. Thus, a putative binding site for a transcription regulator of the Crp/Fnr family, an ArgR-binding site, and two cis-acting catabolite response elements were identified in the promoter-operator region of the operon. Consistent with this, we could demonstrate that the ADS of S. suis is inducible by arginine and reduced O2 tension and subject to carbon catabolite repression. Furthermore, comparing an arcA knockout mutant in which expression of the three operon-encoded proteins was abolished with the parental wild-type strain showed that the arcABC operon of S. suis contributes to survival under acidic conditions.

  3. 30 CFR 285.902 - What are the general requirements for decommissioning for facilities authorized under my SAP, COP...

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... BUREAU OF OCEAN ENERGY MANAGEMENT, REGULATION, AND ENFORCEMENT, DEPARTMENT OF THE INTERIOR OFFSHORE RENEWABLE ENERGY ALTERNATE USES OF EXISTING FACILITIES ON THE OUTER CONTINENTAL SHELF Decommissioning... facilities, projects, cables, pipelines, and obstructions; (2) Clear the seafloor of all obstructions created...

  4. 78 FR 20564 - Medicare and Medicaid Programs; Survey, Certification and Enforcement Procedures

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-04-05

    ... Providers Act of 2008 NF--Nursing Facility OIG--Office of the Inspector General OPT--Provider of outpatient... Act SNF--Skilled Nursing Facility TJC--The Joint Commission I. Background To participate in the... participation (CoPs) for most providers, requirements for skilled nursing facilities (SNFs), conditions for...

  5. Rehabilitation living lab in the mall community of practice: learning together to improve rehabilitation, participation and social inclusion for people living with disabilities.

    PubMed

    Mazer, Barbara; Kairy, Dahlia; Guindon, Andréanne; Girard, Michel; Swaine, Bonnie; Kehayia, Eva; Labbé, Delphine

    2015-04-22

    Communities of practice (CoP) can facilitate collaboration between people who share a common interest, but do not usually work together. A CoP was initiated and developed including stakeholders from clinical, research, community and governmental backgrounds involved in a large multidisciplinary and multi-sectorial project: the Rehabilitation Living Lab in a Mall (RehabMaLL). This study aimed to evaluate the structure, process and outcomes of this CoP. A single case-study, using mixed-methods, evaluated the RehabMaLL CoP initiative after one year, based on Donabedian's conceptual evaluation model. Forty-three participants took part in the RehabMaLL CoP with 60.5% (n = 26) participating at least once on the online platform where 234 comments were posted. Four in-person meetings were held. Members expressed satisfaction regarding the opportunity to share knowledge with people from diverse backgrounds and the usefulness of the CoP for the RehabMaLL project. Collaboration led to concrete outcomes, such as a sensitization activity and a research project. Common challenges included lack of time and difficulty finding common objectives. A CoP can be a useful strategy to facilitate knowledge sharing on disability issues. Future research is necessary to determine strategies of increasing knowledge creation between members.

  6. Acute Positive Effects of Exercise on Center-of-Pressure Fluctuations During Quiet Standing in Middle-Aged and Elderly Women.

    PubMed

    Fukusaki, Chiho; Masani, Kei; Miyasaka, Maya; Nakazawa, Kimitaka

    2016-01-01

    Acute effects of exercise on postural stability have been studied with a focus on fatigue. This study investigated the acute effects of moderate-intensity exercise on center-of-pressure (COP) fluctuation measures in middle-aged and elderly women. Thirty-five healthy women volunteered: 18 women performed a moderate aquatic exercise session for 80 minutes and 17 remained calm in a sitting position for the same duration. Center-of-pressure fluctuations during quiet standing were recorded for 60 seconds with eyes open and closed before and after the exercise and sitting tasks. The time- and frequency-domain measures of the COP time series were calculated. The frequency-domain measures were also calculated for the COP velocity time series. According to 2-way analysis of variance and paired t-tests with a Bonferroni's correction, mean velocity of COP fluctuations, mean velocity of COP fluctuations in the medial-lateral (ML) direction, and total power of the COP velocity time series in the ML direction exhibited significant reductions after 1 session of exercise. These results indicated that a moderate-intensity aquatic exercise decreased COP velocity, counteracting age-related and fatigue-inducing postural deterioration. Therefore, we concluded that a single session of moderate-intensity aquatic exercise has acute positive effects on postural stability in middle-aged and elderly women.

  7. Co3O4/CoP composite hollow polyhedron: A superior catalyst with dramatic efficiency and stability for the room temperature reduction of 4-nitrophenol

    NASA Astrophysics Data System (ADS)

    Liu, Xing; Li, Xiangqing; Qin, Lixia; Mu, Jin; Kang, Shi-Zhao

    2018-03-01

    In the present work, Co3O4/CoP composite hollow polyhedrons were prepared and characterized with X-ray diffraction, scanning electron microscopy, transmission electron microscopy, X-ray photoelectron spectroscopy and N2 adsorption-desorption isotherms. Then, the catalytic activity of the as-prepared Co3O4/CoP hollow polyhedrons was evaluated for the borohydride-assisted reduction of 4-nitrophenol at room temperature. The results indicate that the as-prepared Co3O4/CoP hollow polyhedrons are an efficient recyclable catalyst for the reduction of 4-nitrophenol. When the 4-nitrophenol initial concentration is 1.0 × 10-4 mol L-1 (100 mL), almost 100% 4-nitrophenol can be reduced within 3 min in the presence of the Co3O4/CoP hollow polyhedrons. The apparent rate constant of the 4-nitrophenol reduction is 1.61 min-1 at room temperature, and the activity factor is about 5.37 × 104 mL min-1 g-1, which is almost two times higher than that over Ag nanoparticles. Finally, the catalytic mechanism was preliminarily discussed. It is found that CoP plays an important role in the catalytic process. Here, CoP serves as active sites, which leads to efficient formation of hydrogen atoms from BH4- and fast electron transfer.

  8. Rehabilitation Living Lab in the Mall Community of Practice: Learning Together to Improve Rehabilitation, Participation and Social Inclusion for People Living with Disabilities

    PubMed Central

    Mazer, Barbara; Kairy, Dahlia; Guindon, Andréanne; Girard, Michel; Swaine, Bonnie; Kehayia, Eva; Labbé, Delphine

    2015-01-01

    Communities of practice (CoP) can facilitate collaboration between people who share a common interest, but do not usually work together. A CoP was initiated and developed including stakeholders from clinical, research, community and governmental backgrounds involved in a large multidisciplinary and multi-sectorial project: the Rehabilitation Living Lab in a Mall (RehabMaLL). This study aimed to evaluate the structure, process and outcomes of this CoP. A single case-study, using mixed-methods, evaluated the RehabMaLL CoP initiative after one year, based on Donabedian’s conceptual evaluation model. Forty-three participants took part in the RehabMaLL CoP with 60.5% (n = 26) participating at least once on the online platform where 234 comments were posted. Four in-person meetings were held. Members expressed satisfaction regarding the opportunity to share knowledge with people from diverse backgrounds and the usefulness of the CoP for the RehabMaLL project. Collaboration led to concrete outcomes, such as a sensitization activity and a research project. Common challenges included lack of time and difficulty finding common objectives. A CoP can be a useful strategy to facilitate knowledge sharing on disability issues. Future research is necessary to determine strategies of increasing knowledge creation between members. PMID:25913187

  9. Monodispersepoly[BMA-co-(COPS-I)] Particles by Soap-Free Emulsion Copolymerization and Its Optical Properties as Photonic Crystals.

    PubMed

    Lee, Ki Chang; Choo, Hun Seung

    2015-10-01

    In order to study the surfactant-free emulsion copolymerization of benzyl methacrylate (BMA) with sodium 1-allyloxy-2-hydroxypropane sulfonate (COPS-I) and the resulting optical properties, a series of experiments was carried out at various reaction conditions such as the changes of BMA concentration, COPS-I concentration, BMA concentration under a fixed COPS-I amount, initiator and divinyl benzene (DVB) concentration. All the latices showed highly monodispersed spherical particles in the size range of 144~435 nm and the respective shiny structural colors from their colloidal photonic crystals. It is found that the changes in such polymerization factors greatly affect the number of particles and particle diameter, polymerization rate, molecular weight, zeta-potential, and refractive indices. The increase of number of particles led to the increased rate of polymerization and zeta-potential of the latices, on the other hand, to the decreased molecular weight. Refractive indices and the reflectivity increased with COPS-I concentration, on the other hand, and decreased with DVB concentration. Especially, refractive indices of the resulting poly[BMA-co-(COPS-I)] colloidal photonic crystals showed much higher values of 1.65~2.21 than that of polystyrene, due to the formation of core-shell shaped morphology. Monodisperse and high refractive index of poly[BMA-co-(COPS-I)] particles prepared in this work could be used for the study in photonic crystals and electrophoretic display.

  10. Drosophila COP9 signalosome subunit 7 interacts with multiple genomic loci to regulate development

    PubMed Central

    Singer, Ruth; Atar, Shimshi; Atias, Osnat; Oron, Efrat; Segal, Daniel; Hirsch, Joel A.; Tuller, Tamir; Orian, Amir; Chamovitz, Daniel A.

    2014-01-01

    The COP9 signalosome protein complex has a central role in the regulation of development of multicellular organisms. While the function of this complex in ubiquitin-mediated protein degradation is well established, results over the past few years have hinted that the COP9 signalosome may function more broadly in the regulation of gene expression. Here, using DamID technology, we show that COP9 signalosome subunit 7 functionally associates with a large number of genomic loci in the Drosophila genome, and show that the expression of many genes within these loci is COP9 signalosome-dependent. This association is likely direct as we show CSN7 binds DNA in vitro. The genes targeted by CSN7 are preferentially enriched for transcriptionally active regions of the genome, and are involved in the regulation of distinct gene ontology groupings including imaginal disc development and cell-cycle control. In accord, loss of CSN7 function leads to cell-cycle delay and altered wing development. These results indicate that CSN7, and by extension the entire COP9 signalosome, functions directly in transcriptional control. While the COP9 signalosome protein complex has long been known to regulate protein degradation, here we expand the role of this complex by showing that subunit 7 binds DNA in vitro and functions directly in vivo in transcriptional control of developmentally important pathways that are relevant for human health. PMID:25106867

  11. Fnr, NarP, and NarL Regulation of Escherichia coli K-12 napF (Periplasmic Nitrate Reductase) Operon Transcription In Vitro

    PubMed Central

    Darwin, Andrew J.; Ziegelhoffer, Eva C.; Kiley, Patricia J.; Stewart, Valley

    1998-01-01

    The expression of several Escherichia coli operons is activated by the Fnr protein during anaerobic growth and is further controlled in response to nitrate and nitrite by the homologous response regulators, NarL and NarP. Among these operons, the napF operon, encoding a periplasmic nitrate reductase, has unique features with respect to its Fnr-, NarL-, and NarP-dependent regulation. First, the Fnr-binding site is unusually located compared to the control regions of most other Fnr-activated operons, suggesting different Fnr-RNA polymerase contacts during transcriptional activation. Second, nitrate and nitrite activation is solely dependent on NarP but is antagonized by the NarL protein. In this study, we used DNase I footprint analysis to confirm our previous assignment of the unusual location of the Fnr-binding site in the napF control region. In addition, the in vivo effects of Fnr-positive control mutations on napF operon expression indicate that the napF promoter is atypical with respect to Fnr-mediated activation. The transcriptional regulation of napF was successfully reproduced in vitro by using a supercoiled plasmid template and purified Fnr, NarL, and NarP proteins. These in vitro transcription experiments demonstrate that, in the presence of Fnr, the NarP protein causes efficient transcription activation whereas the NarL protein does not. This suggests that Fnr and NarP may act synergistically to activate napF operon expression. As observed in vivo, this activation by Fnr and NarP is antagonized by the addition of NarL in vitro. PMID:9696769

  12. The Antisense RNA As1_flv4 in the Cyanobacterium Synechocystis sp. PCC 6803 Prevents Premature Expression of the flv4-2 Operon upon Shift in Inorganic Carbon Supply*

    PubMed Central

    Eisenhut, Marion; Georg, Jens; Klähn, Stephan; Sakurai, Isamu; Mustila, Henna; Zhang, Pengpeng; Hess, Wolfgang R.; Aro, Eva-Mari

    2012-01-01

    The functional relevance of natural cis-antisense transcripts is mostly unknown. Here we have characterized the association of three antisense RNAs and one intergenically encoded noncoding RNA with an operon that plays a crucial role in photoprotection of photosystem II under low carbon conditions in the cyanobacterium Synechocystis sp. PCC 6803. Cyanobacteria show strong gene expression dynamics in response to a shift of cells from high carbon to low levels of inorganic carbon (Ci), but the regulatory mechanisms are poorly understood. Among the most up-regulated genes in Synechocystis are flv4, sll0218, and flv2, which are organized in the flv4-2 operon. The flavodiiron proteins encoded by this operon open up an alternative electron transfer route, likely starting from the QB site in photosystem II, under photooxidative stress conditions. Our expression analysis of cells shifted from high carbon to low carbon demonstrated an inversely correlated transcript accumulation of the flv4-2 operon mRNA and one antisense RNA to flv4, designated as As1_flv4. Overexpression of As1_flv4 led to a decrease in flv4-2 mRNA. The promoter activity of as1_flv4 was transiently stimulated by Ci limitation and negatively regulated by the AbrB-like transcription regulator Sll0822, whereas the flv4-2 operon was positively regulated by the transcription factor NdhR. The results indicate that the tightly regulated antisense RNA As1_flv4 establishes a transient threshold for flv4-2 expression in the early phase after a change in Ci conditions. Thus, it prevents unfavorable synthesis of the proteins from the flv4-2 operon. PMID:22854963

  13. Hierarchical CoP/Ni 5 P 4 /CoP microsheet arrays as a robust pH-universal electrocatalyst for efficient hydrogen generation

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

    Mishra, Ishwar Kumar; Zhou, Haiqing; Sun, Jingying

    Exceptional Pt-like electrocatalytic activity was achieved in a sandwich-like catalyst of CoP/Ni 5 P 4 /CoP microsheet arrays for pH-universal hydrogen evolution through simply wrapping Ni 5 P 4 nanosheet arrays in CoP nanoparticles.

  14. Hierarchical CoP/Ni 5 P 4 /CoP microsheet arrays as a robust pH-universal electrocatalyst for efficient hydrogen generation

    DOE PAGES

    Mishra, Ishwar Kumar; Zhou, Haiqing; Sun, Jingying; ...

    2018-01-01

    Exceptional Pt-like electrocatalytic activity was achieved in a sandwich-like catalyst of CoP/Ni 5 P 4 /CoP microsheet arrays for pH-universal hydrogen evolution through simply wrapping Ni 5 P 4 nanosheet arrays in CoP nanoparticles.

  15. Discovery of an operon that participates in agmatine metabolism and regulates biofilm formation in Pseudomonas aeruginosa

    PubMed Central

    Williams, Bryan J.; Du, Rui-Hong; Calcutt, M. Wade; Abdolrasulnia, Rasul; Christman, Brian W.; Blackwell, Timothy S.

    2013-01-01

    Summary Agmatine is the decarboxylation product of arginine and a number of bacteria have devoted enzymatic pathways for its metabolism. Pseudomonas aeruginosa harbours the aguBA operon that metabolizes agmatine to putrescine, which can be subsequently converted into other polyamines or shunted into the TCA cycle for energy production. We discovered an alternate agmatine operon in the P. aeruginosa strain PA14 named agu2ABCA′ that contains two genes for agmatine deiminases (agu2A and agu2A′). This operon was found to be present in 25% of clinical P. aeruginosa isolates. Agu2A′ contains a twin-arginine translocation signal at its N-terminus and site-directed mutagenesis and cell fractionation experiments confirmed this protein is secreted to the periplasm. Analysis of the agu2ABCA′ promoter demonstrates that agmatine induces expression of the operon during the stationary phase of growth and during biofilm growth and agu2ABCA′ provides only weak complementation of aguBA, which is induced during log phase. Biofilm assays of mutants of all three agmatine deiminase genes in PA14 revealed that deletion of agu2ABCA′, specifically its secreted product Agu2A′, reduces biofilm production of PA14 following addition of exogenous agmatine. Together, these findings reveal a novel role for the agu2ABCA′ operon in the biofilm development of P. aeruginosa. PMID:20149107

  16. Delivery of high-energy radiation in midinfrared spectral region by hollow waveguides

    NASA Astrophysics Data System (ADS)

    Nemec, Michal; Jelinkova, Helena; Sulc, Jan; Cerny, Pavel; Miyagi, Mitsunobu; Iwai, Katsumasa; Abe, Yukio; Shi, Yi-Wei; Matsuura, Yuji

    2003-07-01

    Due to increasing number of requirements dealing with the application of a high energy mid-infrared radiation in various branches of medicine (cardiology, dentistry, dermatology, urology, gastroenterology), an enough flexible and lossless delivery system is required. For a transport of this high energy pulses in a mid-infrared region special cyclic olefin polymer-coated silver (COP/Ag) hollow glass waveguides were prepared and tested. A length of the waveguides was 0.5 m and inner diameter 1 mm. As a radiation source, an Er:YAG laser was used. The system generated the energy up to 2.16 J or 2.35 J (in dependence on a repetition rate used - 3 Hz or 4 Hz, respectively). The length of transmitted pulses was measured to be from 110 up to 550 usec in dependence on output energy used. The output radiation was coupled into the COP/Ag waveguide and a throughput and losses values were measured in dependence to input radiation parameters. The transmission obtained was 91%. The maximum delivered energy was dependent on a damage threshold of the waveguide. It was found that the damage threshold is dependent on the repetition rate which shows the dependences on the heat dissipated in the waveguide wall. The value of the damage was 1.7 J and 1.5 J for 3 Hz and 4 Hz repetition rate, respectively. The safe delivered power reached the value of 5 W. The characteristics obtained make this specially constructed COP/Ag hollow glass waveguide promising for the delivery of high-energy laser pulses in medicine and also in other applications.

  17. Ramadan Fasting Effects on Postural Control in the Elderly: A Comparison Between Fallers and Non-fallers.

    PubMed

    Laatar, Rabeb; Baccouch, Rym; Borji, Rihab; Kachouri, Hiba; Rebai, Haithem; Sahli, Sonia

    2016-11-01

    Our purpose was to compare the effects of Ramadan fasting on postural control in elderly fallers and non-fallers. The protocol involved twenty-four healthy old volunteer males divided into two groups: fallers (mean age = 75.43 ± 5.26 years, weight = 67.25 ± 5.30 kg and height = 1.65 ± 0.02 m) and non-fallers (mean age = 72.3 ± 6.42 years, weight = 65.5 ± 6.15 kg and height = 1.64 ± 0.03 m). Participants performed a simple reaction time test (SRT) and a postural control protocol on four different occasions: one week before Ramadan (BR), during the second (SWR) and the fourth week (FWR) of Ramadan and three weeks after Ramadan (AR). Center of pressure (CoP) parameters [the CoP medial-lateral length (CoP X ), and the CoP antero-posterior length (CoP Y )] were assessed using a force platform under two surface conditions: Firm surface and Foam surface. The results showed that Ramadan fasting influences similarly fallers and non-fallers. In fact, for both groups, the CoP X and the CoP Y values increased significantly during the SWR and the FWR compared to BR. These CoP parameters decreased significantly in the Firm surface conditions in the FWR. Moreover, the CoP parameters were significantly higher during the FWR and AR in comparison with BR in the Foam surface conditions. However, the amplitude of increase of the CoP [X] % and the CoP [Y] % (ΔCoP [X] % and ΔCoP [Y] %) between BR and the SWR was significantly higher in the fallers than non-fallers. The SRT of elderly fallers and non-fallers was significantly higher in the SWR and in the FWR compared to BR. In conclusion, Ramadan fasting alters similarly postural control of elderly fallers and non-fallers, but the amplitude (ΔCoP [X] % and ΔCoP [Y] %) of this alteration seems to be more pronounced in fallers than non-fallers. A probable beginning of adaptation occurs at the FWR. However, three weeks seems to be insufficient to recover postural control alterations due to Ramadan fasting in difficult sensory conditions in elderly fallers and non-fallers.

  18. MicroRNA-214 Promotes Apoptosis in Canine Hemangiosarcoma by Targeting the COP1-p53 Axis.

    PubMed

    Heishima, Kazuki; Mori, Takashi; Sakai, Hiroki; Sugito, Nobuhiko; Murakami, Mami; Yamada, Nami; Akao, Yukihiro; Maruo, Kohji

    2015-01-01

    MicroRNA-214 regulates both angiogenic function in endothelial cells and apoptosis in various cancers. However, the regulation and function of miR-214 is unclear in canine hemangiosarcoma, which is a spontaneous model of human angiosarcoma. The expression and functional roles of miR-214 in canine hemangiosarcoma were presently explored by performing miRNA TaqMan qRT-PCR and transfecting cells with synthetic microRNA. Here, we report that miR-214 was significantly down-regulated in the cell lines used and in clinical samples of canine hemangiosarcoma. Restoration of miR-214 expression reduced cell growth and induced apoptosis in canine hemangiosarcoma cell lines through transcriptional activation of p53-regulated genes although miR-214 had a slight effect of growth inhibition on normal endothelial cells. We identified COP1, which is a critical negative regulator of p53, as a novel direct target of miR-214. COP1 was overexpressed and the specific COP1 knockdown induced apoptosis through transcriptional activation of p53-regulated genes as well as did miR-214-transfection in HSA cell lines. Furthermore, p53 knockdown abolished the miR-214-COP1-mediated apoptosis; thus, miR-214 and COP1 regulated apoptosis through controlling p53 in HSA. In conclusion, miR-214 functioned as a tumor suppressor in canine hemangiosarcoma by inducing apoptosis through recovering the function of p53. miR-214 down-regulation and COP1 overexpression is likely to contribute to tumorigenesis of HSA. Therefore, targeting miR-214-COP1-p53 axis would possibly be a novel effective strategy for treatment of canine hemangiosarcoma and capable of being applied to the development of novel therapeutics for human angiosarcoma.

  19. COPS: A Sensitive and Accurate Tool for Detecting Somatic Copy Number Alterations Using Short-Read Sequence Data from Paired Samples

    PubMed Central

    Krishnan, Neeraja M.; Gaur, Prakhar; Chaudhary, Rakshit; Rao, Arjun A.; Panda, Binay

    2012-01-01

    Copy Number Alterations (CNAs) such as deletions and duplications; compose a larger percentage of genetic variations than single nucleotide polymorphisms or other structural variations in cancer genomes that undergo major chromosomal re-arrangements. It is, therefore, imperative to identify cancer-specific somatic copy number alterations (SCNAs), with respect to matched normal tissue, in order to understand their association with the disease. We have devised an accurate, sensitive, and easy-to-use tool, COPS, COpy number using Paired Samples, for detecting SCNAs. We rigorously tested the performance of COPS using short sequence simulated reads at various sizes and coverage of SCNAs, read depths, read lengths and also with real tumor:normal paired samples. We found COPS to perform better in comparison to other known SCNA detection tools for all evaluated parameters, namely, sensitivity (detection of true positives), specificity (detection of false positives) and size accuracy. COPS performed well for sequencing reads of all lengths when used with most upstream read alignment tools. Additionally, by incorporating a downstream boundary segmentation detection tool, the accuracy of SCNA boundaries was further improved. Here, we report an accurate, sensitive and easy to use tool in detecting cancer-specific SCNAs using short-read sequence data. In addition to cancer, COPS can be used for any disease as long as sequence reads from both disease and normal samples from the same individual are available. An added boundary segmentation detection module makes COPS detected SCNA boundaries more specific for the samples studied. COPS is available at ftp://115.119.160.213 with username “cops” and password “cops”. PMID:23110103

  20. Immunogenicity and efficacy following sequential parenterally-administered doses of Salmonella Enteritidis COPS:FliC glycoconjugates in infant and adult mice.

    PubMed

    Baliban, Scott M; Curtis, Brittany; Toema, Deanna; Tennant, Sharon M; Levine, Myron M; Pasetti, Marcela F; Simon, Raphael

    2018-05-23

    In sub-Saharan Africa, invasive nontyphoidal Salmonella (iNTS) infections with serovars S. Enteritidis, S. Typhimurium and I 4,[5],12:i:- are widespread in children < 5 years old. Development of an efficacious vaccine would provide an important public health tool to prevent iNTS disease in this population. Glycoconjugates of S. Enteritidis core and O-polysaccharide (COPS) coupled to the homologous serovar phase 1 flagellin protein (FliC) were previously shown to be immunogenic and protected adult mice against death following challenge with a virulent Malian S. Enteritidis blood isolate. This study extends these observations to immunization of mice in early life and also assesses protection with partial and full regimens. Anti-COPS and anti-FliC serum IgG titers were assessed in infant and adult mice after immunization with 1, 2 or 3 doses of S. Enteritidis COPS:FliC alone or co-formulated with aluminum hydroxide or monophosphoryl lipid A (MPL) adjuvants. S. Enteritidis COPS:FliC was immunogenic in both age groups, although the immune responses were quantitatively lower in infants. Kinetics of antibody production were similar for the native and adjuvanted formulations after three doses; conjugates formulated with MPL elicited significantly increased anti-COPS IgG titers in adult but not infant mice. Nevertheless, robust protection against S. Enteritidis challenge was seen for all three formulations when three doses were given either during infancy or as adults. We further found that significant protection could be achieved with two COPS:FliC doses, despite elicitation of modest serum anti-COPS IgG antibody titers. These findings guide potential immunization strategies that may be translated to develop a human pediatric iNTS vaccine for sub-Saharan Africa.

  1. Estimation of Foot Plantar Center of Pressure Trajectories with Low-Cost Instrumented Insoles Using an Individual-Specific Nonlinear Model.

    PubMed

    Hu, Xinyao; Zhao, Jun; Peng, Dongsheng; Sun, Zhenglong; Qu, Xingda

    2018-02-01

    Postural control is a complex skill based on the interaction of dynamic sensorimotor processes, and can be challenging for people with deficits in sensory functions. The foot plantar center of pressure (COP) has often been used for quantitative assessment of postural control. Previously, the foot plantar COP was mainly measured by force plates or complicated and expensive insole-based measurement systems. Although some low-cost instrumented insoles have been developed, their ability to accurately estimate the foot plantar COP trajectory was not robust. In this study, a novel individual-specific nonlinear model was proposed to estimate the foot plantar COP trajectories with an instrumented insole based on low-cost force sensitive resistors (FSRs). The model coefficients were determined by a least square error approximation algorithm. Model validation was carried out by comparing the estimated COP data with the reference data in a variety of postural control assessment tasks. We also compared our data with the COP trajectories estimated by the previously well accepted weighted mean approach. Comparing with the reference measurements, the average root mean square errors of the COP trajectories of both feet were 2.23 mm (±0.64) (left foot) and 2.72 mm (±0.83) (right foot) along the medial-lateral direction, and 9.17 mm (±1.98) (left foot) and 11.19 mm (±2.98) (right foot) along the anterior-posterior direction. The results are superior to those reported in previous relevant studies, and demonstrate that our proposed approach can be used for accurate foot plantar COP trajectory estimation. This study could provide an inexpensive solution to fall risk assessment in home settings or community healthcare center for the elderly. It has the potential to help prevent future falls in the elderly.

  2. Comparison of Path Length and Ranges of Movement of the Center of Pressure and Reaction Time and Between Paired-Play and Solo-Play of a Virtual Reality Game.

    PubMed

    Portnoy, Sigal; Hersch, Ayelet; Sofer, Tal; Tresser, Sarit

    2017-06-01

    To test whether paired-play will induce longer path length and ranges of movement of the center of pressure (COP), which reflects on balance performance and stability, compared to solo-play and to test the difference in the path length and ranges of movement of the COP while playing the virtual reality (VR) game with the dominant hand compared to playing it with the nondominant hand. In this cross-sectional study 20 children (age 6.1 ± 0.7 years old) played an arm movement controlled VR game alone and with a peer while each of them stood on a pressure measuring pad to track the path length and ranges of movement of the COP. The total COP path was significantly higher during the paired-play (median 295.8 cm) compared to the COP path during the solo-play (median 189.2 cm). No significant differences were found in the reaction time and the mediolateral and anterior-posterior COP ranges between solo-play and paired-play. No significant differences were found between the parameters extracted during paired-play with the dominant or nondominant hand. Our findings imply that the paired-play is advantageous compared to solo-play since it induces a greater movement for the child, during which, higher COP velocities are reached that may contribute to improving the balance control of the child. Apart from the positive social benefits of paired-play, this positive effect on the COP path length is a noteworthy added value in the clinical setting when treating children with balance disorder.

  3. Estimation of Foot Plantar Center of Pressure Trajectories with Low-Cost Instrumented Insoles Using an Individual-Specific Nonlinear Model

    PubMed Central

    Hu, Xinyao; Zhao, Jun; Peng, Dongsheng

    2018-01-01

    Postural control is a complex skill based on the interaction of dynamic sensorimotor processes, and can be challenging for people with deficits in sensory functions. The foot plantar center of pressure (COP) has often been used for quantitative assessment of postural control. Previously, the foot plantar COP was mainly measured by force plates or complicated and expensive insole-based measurement systems. Although some low-cost instrumented insoles have been developed, their ability to accurately estimate the foot plantar COP trajectory was not robust. In this study, a novel individual-specific nonlinear model was proposed to estimate the foot plantar COP trajectories with an instrumented insole based on low-cost force sensitive resistors (FSRs). The model coefficients were determined by a least square error approximation algorithm. Model validation was carried out by comparing the estimated COP data with the reference data in a variety of postural control assessment tasks. We also compared our data with the COP trajectories estimated by the previously well accepted weighted mean approach. Comparing with the reference measurements, the average root mean square errors of the COP trajectories of both feet were 2.23 mm (±0.64) (left foot) and 2.72 mm (±0.83) (right foot) along the medial–lateral direction, and 9.17 mm (±1.98) (left foot) and 11.19 mm (±2.98) (right foot) along the anterior–posterior direction. The results are superior to those reported in previous relevant studies, and demonstrate that our proposed approach can be used for accurate foot plantar COP trajectory estimation. This study could provide an inexpensive solution to fall risk assessment in home settings or community healthcare center for the elderly. It has the potential to help prevent future falls in the elderly. PMID:29389857

  4. Chlamydia pneumoniae effector chlamydial outer protein N sequesters fructose bisphosphate aldolase A, providing a benefit to bacterial growth.

    PubMed

    Ishida, Kasumi; Matsuo, Junji; Yamamoto, Yoshimasa; Yamaguchi, Hiroyuki

    2014-12-21

    Pathogenic chlamydiae are obligate intracellular pathogens and have adapted successfully to human cells, causing sexually transmitted diseases or pneumonia. Chlamydial outer protein N (CopN) is likely a critical effector protein secreted by the type III secretion system in chlamydiae, which manipulates host cells. However, the mechanisms of its action remain to be clarified. In this work, we aimed to identify previously unidentified CopN effector target in host cells. We first performed a pull-down assay with recombinant glutathione S-transferase (GST) fusion CopN proteins (GST-CpCopN: Chlamydia pneumoniae TW183, GST-CtCopN: Chlamydia trachomatis D/UW-3/CX) as "bait" and soluble lysates obtained from human immortal epithelial HEp-2 cells as "prey", followed by SDS-PAGE with mass spectroscopy (MS). We found that a host cell protein specifically bound to GST-CpCopN, but not GST-CtCopN. MS revealed the host protein to be fructose bisphosphate aldolase A (aldolase A), which plays a key role in glycolytic metabolism. We also confirmed the role of aldolase A in chlamydia-infected HEp-2 cells by using two distinct experiments for gene knockdown with an siRNA specific to aldolase A transcripts, and for assessment of glycolytic enzyme gene expression levels. As a result, both the numbers of chlamydial inclusion-forming units and RpoD transcripts were increased in the chlamydia-infected aldolase A knockdown cells, as compared with the wild-type HEp-2 cells. Meanwhile, chlamydial infection tended to enhance expression of aldolase A. We discovered that one of the C. pneumoniae CopN targets is the glycolytic enzyme aldolase A. Sequestering aldolase A may be beneficial to bacterial growth in infected host cells.

  5. Sharing best practices through online communities of practice: a case study.

    PubMed

    Thomas, Annamma Udaya; Fried, Grace P; Johnson, Peter; Stilwell, Barbara J

    2010-11-12

    The USAID-funded Capacity Project established the Global Alliance for Pre-Service Education (GAPS) to provide an online forum to discuss issues related to teaching and acquiring competence in family planning, with a focus on developing countries' health related training institutions. The success of the Global Alliance for Nursing and Midwifery's ongoing web-based community of practice (CoP) provided a strong example of the successful use of this medium to reach many participants in a range of settings. GAPS functioned as a moderated set of forums that were analyzed by a small group of experts in family planning and pre-service education from three organizations. The cost of the program included the effort provided by the moderators and the time to administer responses and conduct the analysis. Family planning is still considered a minor topic in health related training institutions. Rather than focusing solely on family planning competencies, GAPS members suggested a focus on several professional competencies (e.g. communication, leadership, cultural sensitivity, teamwork and problem solving) that would enhance the resulting health care graduate's ability to operate in a complex health environment. Resources to support competency-based education in the academic setting must be sufficient and appropriately distributed. Where clinical competencies are incorporated into pre-service education, responsible faculty and preceptors must be clinically proficient. The interdisciplinary GAPS memberships allowed for a comparison and contrast of competencies, opportunities, promising practices, documents, lessons learned and key teaching strategies. Online CoPs are a useful interface for connecting developing country experiences. From CoPs, we may uncover challenges and opportunities that are faced in the absorption of key public health competencies required for decreasing maternal mortality and morbidity. Use of the World Health Organization (WHO) Implementing Best Practices Knowledge Gateway, which requires only a low bandwidth connection, gave educators an opportunity to engage in the discussion even in the most Internet access-restricted places (e.g. Ethiopia). In order to sustain an online CoP, funds must come from an international organization (e.g. WHO regional office) or university that can program the costs long-term. Eventually, the long-term effectiveness and sustainability of GAPS rests on its transfer to the members themselves.

  6. Attempted suicide by cop: a case study of traumatic brain injury and the insanity defense.

    PubMed

    Bresler, Scott; Scalora, Mario J; Elbogen, Eric B; Moore, Y Scott

    2003-01-01

    Over the years, there have been a number of well-publicized incidents involving persons who seemingly maneuver police officers into shooting them. Such cases, while relatively rare compared with most forms of violence, nevertheless pose difficult challenges to law enforcement agencies. Relatively little is known about persons who engage in suicide by cop incidents. To our knowledge, there has been no published indepth research on instigators of suicide by cop who survived. In this paper, we present a case study of an individual who engaged in and lived through three separate "attempted suicide by cop" incidents. After describing relevant history as well as events of the most recent incident, we compare the case with the extant literature on suicide by cop and analyze commonalities and differences. Finally, we examine the legal considerations involved, with particular attention devoted to the role the individual's traumatic brain injury played in applying the insanity defense.

  7. CORRELATION PURSUIT: FORWARD STEPWISE VARIABLE SELECTION FOR INDEX MODELS

    PubMed Central

    Zhong, Wenxuan; Zhang, Tingting; Zhu, Yu; Liu, Jun S.

    2012-01-01

    In this article, a stepwise procedure, correlation pursuit (COP), is developed for variable selection under the sufficient dimension reduction framework, in which the response variable Y is influenced by the predictors X1, X2, …, Xp through an unknown function of a few linear combinations of them. Unlike linear stepwise regression, COP does not impose a special form of relationship (such as linear) between the response variable and the predictor variables. The COP procedure selects variables that attain the maximum correlation between the transformed response and the linear combination of the variables. Various asymptotic properties of the COP procedure are established, and in particular, its variable selection performance under diverging number of predictors and sample size has been investigated. The excellent empirical performance of the COP procedure in comparison with existing methods are demonstrated by both extensive simulation studies and a real example in functional genomics. PMID:23243388

  8. Continuous punishment and the potential of gentle rule enforcement.

    PubMed

    Erev, Ido; Ingram, Paul; Raz, Ornit; Shany, Dror

    2010-05-01

    The paper explores the conditions that determine the effect of rule enforcement policies that imply an attempt to punish all the visible violations of the rule. We start with a simple game-theoretic analysis that highlights the value of gentle COntinuous Punishment (gentle COP) policies. If the subjects of the rule are rational, gentle COP can eliminate violations even when the rule enforcer has limited resources. The second part of the paper uses simulations to examine the robustness of gentle COP policies to likely deviations from rationality. The results suggest that when the probability of detecting violations is sufficiently high, gentle COP policies can be effective even when the subjects of the rule are boundedly rational adaptive learners. The paper concludes with experimental studies that clarify the value of gentle COP policies in the lab, and in attempt to eliminate cheating in exams. Copyright (c) 2009 Elsevier B.V. All rights reserved.

  9. The chemotaxis regulator pilG of Xylella fastidiosa is required for virulence in Vitis vinifera grapevines

    USDA-ARS?s Scientific Manuscript database

    Xylella fastidiosa is a Gram-negative, xylem-limited pathogenic bacterium that causes Pierce’s disease of grapevines. Type IV pili of X. fastidiosa are regulated by pilG, a chemotaxis regulator in Pil-Chp operon involving signal transduction pathways. To elucidate the role of pilG in twitching motil...

  10. A Eukaryotic-Type Serine/Threonine Protein Kinase Is Required for Biofilm Formation, Genetic Competence, and Acid Resistance in Streptococcus mutans

    PubMed Central

    Hussain, Haitham; Branny, Pavel; Allan, Elaine

    2006-01-01

    We report an operon encoding a eukaryotic-type serine/threonine protein kinase (STPK) and its cognate phosphatase (STPP) in Streptococcus mutans. Mutation of the gene encoding the STPK produced defects in biofilm formation, genetic competence, and acid resistance, determinants important in caries pathogenesis. PMID:16452447

  11. The transcriptional terminator sequences downstream of the covR gene terminate covR/S operon transcription to generate covR monocistronic transcripts in Streptococcus pyogenes.

    PubMed

    Chiang-Ni, Chuan; Tsou, Chih-Cheng; Lin, Yee-Shin; Chuang, Woei-Jer; Lin, Ming-T; Liu, Ching-Chuan; Wu, Jiunn-Jong

    2008-12-31

    CovR/S is an important two component regulatory system, which regulates about 15% of the gene expression in Streptococcus pyogenes. The covR/S locus was identified as an operon generating an RNA transcript around 2.5-kb in size. In this study, we found the covR/S operon produced three RNA transcripts (around 2.5-, 1.0-, and 0.8-kb in size). Using RNA transcriptional terminator sequence prediction and transcriptional terminator analysis, we identified two atypical rho-independent terminator sequences downstream of the covR gene and showed these terminator sequences terminate RNA transcription efficiently. These results indicate that covR/S operon generates covR/S transcript and monocistronic covR transcripts.

  12. A super-family of transcriptional activators regulates bacteriophage packaging and lysis in Gram-positive bacteria

    PubMed Central

    Quiles-Puchalt, Nuria; Tormo-Más, María Ángeles; Campoy, Susana; Toledo-Arana, Alejandro; Monedero, Vicente; Lasa, Íñigo; Novick, Richard P.; Christie, Gail E.; Penadés, José R.

    2013-01-01

    The propagation of bacteriophages and other mobile genetic elements requires exploitation of the phage mechanisms involved in virion assembly and DNA packaging. Here, we identified and characterized four different families of phage-encoded proteins that function as activators required for transcription of the late operons (morphogenetic and lysis genes) in a large group of phages infecting Gram-positive bacteria. These regulators constitute a super-family of proteins, here named late transcriptional regulators (Ltr), which share common structural, biochemical and functional characteristics and are unique to this group of phages. They are all small basic proteins, encoded by genes present at the end of the early gene cluster in their respective phage genomes and expressed under cI repressor control. To control expression of the late operon, the Ltr proteins bind to a DNA repeat region situated upstream of the terS gene, activating its transcription. This involves the C-terminal part of the Ltr proteins, which control specificity for the DNA repeat region. Finally, we show that the Ltr proteins are the only phage-encoded proteins required for the activation of the packaging and lysis modules. In summary, we provide evidence that phage packaging and lysis is a conserved mechanism in Siphoviridae infecting a wide variety of Gram-positive bacteria. PMID:23771138

  13. Good CoPs and Bad CoPs: Facilitating Reform in First-Year Assessment via a Community of Practice

    ERIC Educational Resources Information Center

    Howlett, Catherine; Arthur, James Michael; Ferreira, Jo Anne

    2016-01-01

    The idea and implementation of learning communities are gaining favour in higher education institutions. In particular, there are a number of successful examples to emerge of the application of the Community of Practice (CoP) framework proposed by Wenger [(1998). "Communities of practice: Learning, meaning, and identity." Cambridge, MA:…

  14. 30 CFR 585.714 - What records relating to SAPs, COPs, and GAPs must I keep?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 30 Mineral Resources 2 2014-07-01 2014-07-01 false What records relating to SAPs, COPs, and GAPs must I keep? 585.714 Section 585.714 Mineral Resources BUREAU OF OCEAN ENERGY MANAGEMENT, DEPARTMENT OF... records relating to SAPs, COPs, and GAPs must I keep? (a) Until BOEM releases your financial assurance...

  15. 30 CFR 585.714 - What records relating to SAPs, COPs, and GAPs must I keep?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 30 Mineral Resources 2 2013-07-01 2013-07-01 false What records relating to SAPs, COPs, and GAPs must I keep? 585.714 Section 585.714 Mineral Resources BUREAU OF OCEAN ENERGY MANAGEMENT, DEPARTMENT OF... records relating to SAPs, COPs, and GAPs must I keep? (a) Until BOEM releases your financial assurance...

  16. 30 CFR 585.714 - What records relating to SAPs, COPs, and GAPs must I keep?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 30 Mineral Resources 2 2012-07-01 2012-07-01 false What records relating to SAPs, COPs, and GAPs must I keep? 585.714 Section 585.714 Mineral Resources BUREAU OF OCEAN ENERGY MANAGEMENT, DEPARTMENT OF... records relating to SAPs, COPs, and GAPs must I keep? (a) Until BOEM releases your financial assurance...

  17. 30 CFR 285.714 - What records relating to SAPs, COPs, and GAPs must I keep?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 30 Mineral Resources 2 2011-07-01 2011-07-01 false What records relating to SAPs, COPs, and GAPs must I keep? 285.714 Section 285.714 Mineral Resources BUREAU OF OCEAN ENERGY MANAGEMENT, REGULATION... Verification Agent § 285.714 What records relating to SAPs, COPs, and GAPs must I keep? (a) Until MMS releases...

  18. 20 CFR 10.216 - How is the pay rate for COP calculated?

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... for COP purposes is equal to the employee's regular “weekly” pay (the average of the weekly pay over... occurred during the 45-day period are to be reflected in the weekly pay determination. (b) The weekly pay... 20 Employees' Benefits 1 2010-04-01 2010-04-01 false How is the pay rate for COP calculated? 10...

  19. A Community of Practice Focused on Resiliency in Graduate Nursing Students

    ERIC Educational Resources Information Center

    Wildes, Megan

    2016-01-01

    The purpose of this project was to create a Community of Practice (CoP) focused on resiliency in graduate nursing students. CoPs are networks of people who collectively learn and share in learning as a social experience. By engaging a CoP that focused on resiliency in graduate nursing students, the aim was to positively support students' sense of…

  20. 50 CFR 23.88 - What are the resolutions and decisions of the CoP?

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 50 Wildlife and Fisheries 9 2012-10-01 2012-10-01 false What are the resolutions and decisions of the CoP? 23.88 Section 23.88 Wildlife and Fisheries UNITED STATES FISH AND WILDLIFE SERVICE... decisions of the CoP? (a) Purpose. Under Article XI of the Treaty, the Parties agree to resolutions and...

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