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Sample records for kai semento gijutsu

  1. Circadian oscillations of KaiA-KaiC and KaiB-KaiC complex formations in an in vitro reconstituted KaiABC clock oscillator.

    PubMed

    Murakami, Reiko; Mutoh, Risa; Ishii, Ketaro; Ishiura, Masahiro

    2016-08-01

    The circadian clock is an endogenous biological mechanism that generates autonomous daily cycles in physiological activities. The phosphorylation levels of KaiC oscillated with a period of 24 h in an ATP-dependent clock oscillator reconstituted in vitro from KaiA, KaiB and KaiC. We examined the complex formations of KaiA and KaiB with KaiC in the KaiABC clock oscillator by fluorescence correlation spectrometry (FCS) analysis. The formation of KaiB-containing protein complex(es) oscillated in a circadian manner, with a single peak at 12 h and single trough at 24 h in the circadian cycle, whereas that of KaiA-containing protein complex(es) oscillated with two peaks at 12 and 24 h. FCS and surface plasmon resonance analyses showed that the binding affinity of KaiA for a mutant KaiC with Ala substitutions at the two phosphorylation sites considered to mimic the nonphosphorylated form of KaiC (np-KaiC) was higher than that for a mutant KaiC with Asp substitutions at the two phosphorylation sites considered to mimic the completely phosphorylated form of KaiC (cp-KaiC). The results from the study suggest that a KaiA-KaiB-cp-KaiC ternary complex and a KaiA-np-KaiC complex were formed at 12 and 24 h, respectively. PMID:27477077

  2. Insight into cyanobacterial circadian timing from structural details of the KaiB–KaiC interaction

    PubMed Central

    Snijder, Joost; Burnley, Rebecca J.; Wiegard, Anika; Melquiond, Adrien S. J.; Bonvin, Alexandre M. J. J.; Axmann, Ilka M.; Heck, Albert J. R.

    2014-01-01

    Circadian timing in cyanobacteria is determined by the Kai system consisting of KaiA, KaiB, and KaiC. Interactions between Kai proteins change the phosphorylation status of KaiC, defining the phase of circadian timing. The KaiC–KaiB interaction is crucial for the circadian rhythm to enter the dephosphorylation phase but it is not well understood. Using mass spectrometry to characterize Kai complexes, we found that KaiB forms monomers, dimers, and tetramers. The monomer is the unit that interacts with KaiC, with six KaiB monomers binding to one KaiC hexamer. Hydrogen–deuterium exchange MS reveals structural changes in KaiC upon binding of KaiB in both the CI and CII domains, showing allosteric coupling upon KaiB binding. Based on this information we propose a model of the KaiB–KaiC complex and hypothesize that the allosteric changes observed upon complex formation relate to coupling KaiC ATPase activity with KaiB binding and to sequestration of KaiA dimers into KaiCBA complexes. PMID:24474762

  3. Insight into cyanobacterial circadian timing from structural details of the KaiB-KaiC interaction.

    PubMed

    Snijder, Joost; Burnley, Rebecca J; Wiegard, Anika; Melquiond, Adrien S J; Bonvin, Alexandre M J J; Axmann, Ilka M; Heck, Albert J R

    2014-01-28

    Circadian timing in cyanobacteria is determined by the Kai system consisting of KaiA, KaiB, and KaiC. Interactions between Kai proteins change the phosphorylation status of KaiC, defining the phase of circadian timing. The KaiC-KaiB interaction is crucial for the circadian rhythm to enter the dephosphorylation phase but it is not well understood. Using mass spectrometry to characterize Kai complexes, we found that KaiB forms monomers, dimers, and tetramers. The monomer is the unit that interacts with KaiC, with six KaiB monomers binding to one KaiC hexamer. Hydrogen-deuterium exchange MS reveals structural changes in KaiC upon binding of KaiB in both the CI and CII domains, showing allosteric coupling upon KaiB binding. Based on this information we propose a model of the KaiB-KaiC complex and hypothesize that the allosteric changes observed upon complex formation relate to coupling KaiC ATPase activity with KaiB binding and to sequestration of KaiA dimers into KaiCBA complexes. PMID:24474762

  4. Structural model of the circadian clock KaiB-KaiC complex and mechanism for modulation of KaiC phosphorylation

    SciTech Connect

    Pattanayek, Rekha; Williams, Dewight R; Pattanayek, Sabuj; Mori, Tetsuya; Johnson, Carl H; Stewart, Phoebe L; Egli, Martin

    2010-03-08

    The circadian clock of the cyanobacterium Synechococcus elongatus can be reconstituted in vitro by the KaiA, KaiB and KaiC proteins in the presence of ATP. The principal clock component, KaiC, undergoes regular cycles between hyper- and hypo-phosphorylated states with a period of ca. 24 h that is temperature compensated. KaiA enhances KaiC phosphorylation and this enhancement is antagonized by KaiB. Throughout the cycle Kai proteins interact in a dynamic manner to form complexes of different composition. We present a three-dimensional model of the S. elongatus KaiB-KaiC complex based on X-ray crystallography, negative-stain and cryo-electron microscopy, native gel electrophoresis and modelling techniques. We provide experimental evidence that KaiB dimers interact with KaiC from the same side as KaiA and for a conformational rearrangement of the C-terminal regions of KaiC subunits. The enlarged central channel and thus KaiC subunit separation in the C-terminal ring of the hexamer is consistent with KaiC subunit exchange during the dephosphorylation phase. The proposed binding mode of KaiB explains the observation of simultaneous binding of KaiA and KaiB to KaiC, and provides insight into the mechanism of KaiB's antagonism of KaiA.

  5. Nature of KaiB-KaiC binding in the cyanobacterial circadian oscillator.

    PubMed

    Pattanayek, Rekha; Yadagiri, Kirthi Kiran; Ohi, Melanie D; Egli, Martin

    2013-03-01

    In the cyanobacteria Synechococcus elongatus and Thermosynechococcus elongatus, the KaiA, KaiB and KaiC proteins in the presence of ATP generate a post-translational oscillator (PTO) that can be reconstituted in vitro. KaiC is the result of a gene duplication and resembles a double doughnut with N-terminal CI and C-terminal CII hexameric rings. Six ATPs are bound between subunits in both the CI and CII ring. CI harbors ATPase activity, and CII catalyzes phosphorylation and dephosphorylation at T432 and S431 with a ca. 24-h period. KaiA stimulates KaiC phosphorylation, and KaiB promotes KaiC subunit exchange and sequesters KaiA on the KaiB-KaiC interface in the final stage of the clock cycle. Studies of the PTO protein-protein interactions are convergent in terms of KaiA binding to CII but have led to two opposing models of the KaiB-KaiC interaction. Electron microscopy (EM) and small angle X-ray scattering (SAXS), together with native PAGE using full-length proteins and separate CI and CII rings, are consistent with binding of KaiB to CII. Conversely, NMR together with gel filtration chromatography and denatured PAGE using monomeric CI and CII domains support KaiB binding to CI. To resolve the existing controversy, we studied complexes between KaiB and gold-labeled, full-length KaiC with negative stain EM. The EM data clearly demonstrate that KaiB contacts the CII ring. Together with the outcomes of previous analyses, our work establishes that only CII participates in interactions with KaiA and KaiB as well as with the His kinase SasA involved in the clock output pathway. PMID:23388462

  6. Nature of KaiB-KaiC binding in the cyanobacterial circadian oscillator

    PubMed Central

    Pattanayek, Rekha; Yadagiri, Kirthi Kiran; Ohi, Melanie D.; Egli, Martin

    2013-01-01

    In the cyanobacteria Synechococcus elongatus and Thermosynechococcus elongatus, the KaiA, KaiB and KaiC proteins in the presence of ATP generate a post-translational oscillator (PTO) that can be reconstituted in vitro. KaiC is the result of a gene duplication and resembles a double doughnut with N-terminal CI and C-terminal CII hexameric rings. Six ATPs are bound between subunits in both the CI and CII ring. CI harbors ATPase activity, and CII catalyzes phosphorylation and dephosphorylation at T432 and S431 with a ca. 24-h period. KaiA stimulates KaiC phosphorylation, and KaiB promotes KaiC subunit exchange and sequesters KaiA on the KaiB-KaiC interface in the final stage of the clock cycle. Studies of the PTO protein-protein interactions are convergent in terms of KaiA binding to CII but have led to two opposing models of the KaiB-KaiC interaction. Electron microscopy (EM) and small angle X-ray scattering (SAXS), together with native PAGE using full-length proteins and separate CI and CII rings, are consistent with binding of KaiB to CII. Conversely, NMR together with gel filtration chromatography and denatured PAGE using monomeric CI and CII domains support KaiB binding to CI. To resolve the existing controversy, we studied complexes between KaiB and gold-labeled, full-length KaiC with negative stain EM. The EM data clearly demonstrate that KaiB contacts the CII ring. Together with the outcomes of previous analyses, our work establishes that only CII participates in interactions with KaiA and KaiB as well as with the His kinase SasA involved in the clock output pathway. PMID:23388462

  7. Recent cyanobacterial Kai protein structures suggest a rotary clock.

    PubMed

    Wang, Jimin

    2005-05-01

    The cyanobacterial circadian oscillator consists of three Kai proteins, KaiA, KaiB, and KaiC, in its oscillation feedback loop. Structural comparison reveals that the Kai system resembles the F1-ATPase system in which KaiC is equivalent to alpha(3)beta(3), KaiA to gammadelta, and KaiB to its inhibitory factor. It also suggests that there exists a possible haemagglutinin-like spring-loaded mechanism for the activation of KaiA during the formation of Kai complexes. PMID:15893664

  8. CryoEM and Molecular Dynamics of the Circadian KaiB–KaiC Complex Indicates KaiB Monomers Interact with KaiC and Block ATP Binding Clefts

    PubMed Central

    Villarreal, Seth A.; Pattanayek, Rekha; Williams, Dewight R.; Mori, Tetsuya; Qin, Ximing; Johnson, Carl H.; Egli, Martin; Stewart, Phoebe L.

    2013-01-01

    The circadian control of cellular processes in cyanobacteria is regulated by a posttranslational oscillator formed by three Kai proteins. During the oscillator cycle KaiA serves to promote autophosphorylation of KaiC while KaiB counteracts this effect. Here we present a crystallographic structure of the wild-type Synechococcus elongatus KaiB and a cryoEM structure of a KaiBC complex. The crystal structure shows the expected dimer core structure and significant conformational variations of the KaiB C-terminal region, which is functionally important in maintaining rhythmicity. The KaiBC sample was formed with a C-terminally truncated form of KaiC, KaiC-Δ489, which is persistently phosphorylated. The KaiB-KaiC-Δ489 structure reveals that the KaiC hexamer can bind six monomers of KaiB, which form a continuous ring of density in the KaiBC complex. We performed cryoEM guided molecular dynamics flexible fitting simulations with crystal structures of KaiB and KaiC to probe the KaiBC protein-protein interface. This analysis indicated a favorable binding mode for the KaiB monomer on the CII end of KaiC, involving two adjacent KaiC subunits and spanning an ATP binding cleft. A KaiC mutation, R468C, which has been shown to affect the affinity of KaiB for KaiC and lengthen the period in a bioluminescence rhythm assay, is found within the middle of the predicted KaiBC interface. The proposed KaiB binding mode blocks access to the ATP binding cleft in the CII ring of KaiC, which provides insight into how KaiB might influence the phosphorylation status of KaiC. PMID:23796516

  9. CryoEM and Molecular Dynamics of the Circadian KaiB-KaiC Complex Indicates That KaiB Monomers Interact with KaiC and Block ATP Binding Clefts

    SciTech Connect

    Villarreal, Seth A.; Pattanayek, Rekha; Williams, Dewight R.; Mori, Tetsuya; Qin, Ximing; Johnson, Carl H.; Egli, Martin; Stewart, Phoebe L.

    2014-10-02

    The circadian control of cellular processes in cyanobacteria is regulated by a posttranslational oscillator formed by three Kai proteins. During the oscillator cycle, KaiA serves to promote autophosphorylation of KaiC while KaiB counteracts this effect. Here, we present a crystallographic structure of the wild-type Synechococcus elongatus KaiB and a cryo-electron microscopy (cryoEM) structure of a KaiBC complex. The crystal structure shows the expected dimer core structure and significant conformational variations of the KaiB C-terminal region, which is functionally important in maintaining rhythmicity. The KaiBC sample was formed with a C-terminally truncated form of KaiC, KaiC-Δ489, which is persistently phosphorylated. The KaiB–KaiC-Δ489 structure reveals that the KaiC hexamer can bind six monomers of KaiB, which form a continuous ring of density in the KaiBC complex. We performed cryoEM-guided molecular dynamics flexible fitting simulations with crystal structures of KaiB and KaiC to probe the KaiBC protein–protein interface. This analysis indicated a favorable binding mode for the KaiB monomer on the CII end of KaiC, involving two adjacent KaiC subunits and spanning an ATP binding cleft. A KaiC mutation, R468C, which has been shown to affect the affinity of KaiB for KaiC and lengthen the period in a bioluminescence rhythm assay, is found within the middle of the predicted KaiBC interface. The proposed KaiB binding mode blocks access to the ATP binding cleft in the CII ring of KaiC, which provides insight into how KaiB might influence the phosphorylation status of KaiC.

  10. The ATP-mediated regulation of KaiB-KaiC interaction in the cyanobacterial circadian clock.

    PubMed

    Mutoh, Risa; Nishimura, Atsuhito; Yasui, So; Onai, Kiyoshi; Ishiura, Masahiro

    2013-01-01

    The cyanobacterial circadian clock oscillator is composed of three clock proteins--KaiA, KaiB, and KaiC, and interactions among the three Kai proteins generate clock oscillation in vitro. However, the regulation of these interactions remains to be solved. Here, we demonstrated that ATP regulates formation of the KaiB-KaiC complex. In the absence of ATP, KaiC was monomeric (KaiC(1mer)) and formed a complex with KaiB. The addition of ATP plus Mg(2+) (Mg-ATP), but not that of ATP only, to the KaiB-KaiC(1mer) complex induced the hexamerization of KaiC and the concomitant release of KaiB from the KaiB-KaiC(1mer) complex, indicating that Mg-ATP and KaiB compete each other for KaiC. In the presence of ATP and Mg(2+) (Mg-ATP), KaiC became a homohexameric ATPase (KaiC(6mer)) with bound Mg-ATP and formed a complex with KaiB, but KaiC hexamerized by unhydrolyzable substrates such as ATP and Mg-ATP analogs, did not. A KaiC N-terminal domain protein, but not its C-terminal one, formed a complex with KaiB, indicating that KaiC associates with KaiB via its N-terminal domain. A mutant KaiC(6mer) lacking N-terminal ATPase activity did not form a complex with KaiB whereas a mutant lacking C-terminal ATPase activity did. Thus, the N-terminal domain of KaiC is responsible for formation of the KaiB-KaiC complex, and the hydrolysis of the ATP bound to N-terminal ATPase motifs on KaiC(6mer) is required for formation of the KaiB-KaiC(6mer) complex. KaiC(6mer) that had been hexamerized with ADP plus aluminum fluoride, which are considered to mimic ADP-Pi state, formed a complex with KaiB, suggesting that KaiB is able to associate with KaiC(6mer) with bound ADP-Pi. PMID:24244649

  11. The ATP-Mediated Regulation of KaiB-KaiC Interaction in the Cyanobacterial Circadian Clock

    PubMed Central

    Yasui, So; Onai, Kiyoshi; Ishiura, Masahiro

    2013-01-01

    The cyanobacterial circadian clock oscillator is composed of three clock proteins—KaiA, KaiB, and KaiC, and interactions among the three Kai proteins generate clock oscillation in vitro. However, the regulation of these interactions remains to be solved. Here, we demonstrated that ATP regulates formation of the KaiB-KaiC complex. In the absence of ATP, KaiC was monomeric (KaiC1mer) and formed a complex with KaiB. The addition of ATP plus Mg2+ (Mg-ATP), but not that of ATP only, to the KaiB-KaiC1mer complex induced the hexamerization of KaiC and the concomitant release of KaiB from the KaiB-KaiC1mer complex, indicating that Mg-ATP and KaiB compete each other for KaiC. In the presence of ATP and Mg2+ (Mg-ATP), KaiC became a homohexameric ATPase (KaiC6mer) with bound Mg-ATP and formed a complex with KaiB, but KaiC hexamerized by unhydrolyzable substrates such as ATP and Mg-ATP analogs, did not. A KaiC N-terminal domain protein, but not its C-terminal one, formed a complex with KaiB, indicating that KaiC associates with KaiB via its N-terminal domain. A mutant KaiC6mer lacking N-terminal ATPase activity did not form a complex with KaiB whereas a mutant lacking C-terminal ATPase activity did. Thus, the N-terminal domain of KaiC is responsible for formation of the KaiB-KaiC complex, and the hydrolysis of the ATP bound to N-terminal ATPase motifs on KaiC6mer is required for formation of the KaiB-KaiC6mer complex. KaiC6mer that had been hexamerized with ADP plus aluminum fluoride, which are considered to mimic ADP-Pi state, formed a complex with KaiB, suggesting that KaiB is able to associate with KaiC6mer with bound ADP-Pi. PMID:24244649

  12. Loop-Loop Interactions Regulate KaiA-Stimulated KaiC Phosphorylation in the Cyanobacterial KaiABC Circadian Clock

    SciTech Connect

    Egli, Martin; Pattanayek, Rekha; Sheehan, Jonathan H.; Xu, Yao; Mori, Tetsuya; Smith, Jarrod A.; Johnson, Carl H.

    2013-01-25

    We found that the Synechococcus elongatus KaiA, KaiB, and KaiC proteins in the presence of ATP generate a post-translational oscillator that runs in a temperature-compensated manner with a period of 24 h. KaiA dimer stimulates phosphorylation of KaiC hexamer at two sites per subunit, T432 and S431, and KaiB dimers antagonize KaiA action and induce KaiC subunit exchange. Neither the mechanism of KaiA-stimulated KaiC phosphorylation nor that of KaiB-mediated KaiC dephosphorylation is understood in detail at present. We demonstrate here that the A422V KaiC mutant sheds light on the former mechanism. It was previously reported that A422V is less sensitive to dark pulse-induced phase resetting and has a reduced amplitude of the KaiC phosphorylation rhythm in vivo. A422 maps to a loop (422-loop) that continues toward the phosphorylation sites. By pulling on the C-terminal peptide of KaiC (A-loop), KaiA removes restraints from the adjacent 422-loop whose increased flexibility indirectly promotes kinase activity. We found in the crystal structure that A422V KaiC lacks phosphorylation at S431 and exhibits a subtle, local conformational change relative to wild-type KaiC. Molecular dynamics simulations indicate higher mobility of the 422-loop in the absence of the A-loop and mobility differences in other areas associated with phosphorylation activity between wild-type and mutant KaiCs. Finally, the A-loop–422-loop relay that informs KaiC phosphorylation sites of KaiA dimer binding propagates to loops from neighboring KaiC subunits, thus providing support for a concerted allosteric mechanism of phosphorylation.

  13. Loop-Loop Interactions Regulate KaiA-Stimulated KaiC Phosphorylation in the Cyanobacterial KaiABC Circadian Clock

    PubMed Central

    Egli, Martin; Pattanayek, Rekha; Sheehan, Jonathan H.; Xu, Yao; Mori, Tetsuya; Smith, Jarrod A.; Johnson, Carl H.

    2013-01-01

    The Synechococcus elongatus KaiA, KaiB and KaiC proteins in the presence of ATP generate a post-translational oscillator (PTO) that runs in a temperature-compensated manner with a period of 24 hours. KaiA dimer stimulates phosphorylation of KaiC hexamer at two sites per subunit, T432 and S431, and KaiB dimers antagonize KaiA action and induce KaiC subunit exchange. Neither the mechanism of KaiA-stimulated KaiC phosphorylation nor that of KaiB-mediated KaiC dephosphorylation is understood in detail at the present time. We demonstrate here that the A422V KaiC mutant sheds light on the former mechanism. It was previously reported that A422V is less sensitive to dark pulse-induced phase resetting and has a reduced amplitude of the KaiC phosphorylation rhythm in vivo. A422 maps to a loop (422-loop) that continues toward the phosphorylation sites. By pulling on the C-terminal peptide of KaiC (A-loop), KaiA removes restraints from the adjacent 422-loop whose increased flexibility indirectly promotes kinase activity. We found in the crystal structure that A422V KaiC lacks phosphorylation at S431 and exhibits a subtle, local conformational change relative to wild-type KaiC. MD simulations indicate higher mobility of the 422-loop in the absence of the A-loop and mobility differences in other areas associated with phosphorylation activity between wild-type and mutant KaiCs. The A-loop···422-loop relay that informs KaiC phosphorylation sites of KaiA dimer binding propagates to loops from neighboring KaiC subunits, thus providing support for a concerted allosteric mechanism of phosphorylation. PMID:23351065

  14. A dynamic interaction process between KaiA and KaiC is critical to the cyanobacterial circadian oscillator

    PubMed Central

    Dong, Pei; Fan, Ying; Sun, Jianqiang; Lv, Mengting; Yi, Ming; Tan, Xiao; Liu, Sen

    2016-01-01

    The core circadian oscillator of cyanobacteria consists of three proteins, KaiA, KaiB, and KaiC. This circadian oscillator could be functionally reconstituted in vitro with these three proteins, and therefore has been a very important model in circadian rhythm research. KaiA can bind to KaiC and then stimulate its phosphorylation, but their interaction mechanism remains elusive. In this study, we followed the “second-site suppressor” strategy to investigate the interaction mechanism of KaiA and KaiC. Using protein sequence analyses, we showed that there exist co-varying residues in the binding interface of KaiA and KaiC. The followed mutagenesis study verified that these residues are important to the functions of KaiA and KaiC, but their roles could not be fully explained by the reported complex structures of KaiA and KaiC derived peptides. Combining our data with previous reports, we suggested a dynamic interaction mechanism in KaiA-KaiC interaction, in which both KaiA and the intrinsically disordered tail of KaiC undergo significant structural changes through conformational selection and induced fit during the binding process. At last, we presented a mathematic model to support this hypothesis and explained the importance of this interaction mechanism for the KaiABC circadian oscillator. PMID:27113386

  15. A dynamic interaction process between KaiA and KaiC is critical to the cyanobacterial circadian oscillator.

    PubMed

    Dong, Pei; Fan, Ying; Sun, Jianqiang; Lv, Mengting; Yi, Ming; Tan, Xiao; Liu, Sen

    2016-01-01

    The core circadian oscillator of cyanobacteria consists of three proteins, KaiA, KaiB, and KaiC. This circadian oscillator could be functionally reconstituted in vitro with these three proteins, and therefore has been a very important model in circadian rhythm research. KaiA can bind to KaiC and then stimulate its phosphorylation, but their interaction mechanism remains elusive. In this study, we followed the "second-site suppressor" strategy to investigate the interaction mechanism of KaiA and KaiC. Using protein sequence analyses, we showed that there exist co-varying residues in the binding interface of KaiA and KaiC. The followed mutagenesis study verified that these residues are important to the functions of KaiA and KaiC, but their roles could not be fully explained by the reported complex structures of KaiA and KaiC derived peptides. Combining our data with previous reports, we suggested a dynamic interaction mechanism in KaiA-KaiC interaction, in which both KaiA and the intrinsically disordered tail of KaiC undergo significant structural changes through conformational selection and induced fit during the binding process. At last, we presented a mathematic model to support this hypothesis and explained the importance of this interaction mechanism for the KaiABC circadian oscillator. PMID:27113386

  16. Effects of adenylates on the circadian interaction of KaiB with the KaiC complex in the reconstituted cyanobacterial Kai protein oscillator.

    PubMed

    Goda, Kazuhito; Kondo, Takao; Oyama, Tokitaka

    2014-01-01

    Cyanobacteria are photosynthetic prokaryotes that possess circadian oscillators. Clock proteins, KaiA, KaiB, KaiC compose the central circadian oscillator, which can be reconstituted in vitro in the presence of ATP. KaiC has ATPase, autokinase, and autophosphatase enzymatic activities. These activities are modulated by protein-protein interactions among the Kai proteins. The interaction of KaiB with the KaiC complex shows a circadian rhythm in the reconstituted system. We previously developed a quantitative, real-time monitoring system for the dynamic behavior of the complex using fluorescence correlation spectroscopy. Here, we examined the effects of ATP and ADP on the rhythmic interaction of KaiB. We show that increased concentration of ATP or ADP shortened period length. Adding ADP to the Kai protein oscillation shifted its phase in a phase-dependent manner. These results provide insight into how circadian oscillation entrainment mechanism is linked to cellular metabolism. PMID:25105527

  17. Intermolecular associations determine the dynamics of the circadian KaiABC oscillator

    PubMed Central

    Qin, Ximing; Byrne, Mark; Mori, Tetsuya; Zou, Ping; Williams, Dewight R.; Mchaourab, Hassane; Johnson, Carl Hirschie

    2010-01-01

    Three proteins from cyanobacteria (KaiA, KaiB, and KaiC) can reconstitute circadian oscillations in vitro. At least three molecular properties oscillate during this reaction, namely rhythmic phosphorylation of KaiC, ATP hydrolytic activity of KaiC, and assembly/disassembly of intermolecular complexes among KaiA, KaiB, and KaiC. We found that the intermolecular associations determine key dynamic properties of this in vitro oscillator. For example, mutations within KaiB that alter the rates of binding of KaiB to KaiC also predictably modulate the period of the oscillator. Moreover, we show that KaiA can bind stably to complexes of KaiB and hyperphosphorylated KaiC. Modeling simulations indicate that the function of this binding of KaiA to the KaiB•KaiC complex is to inactivate KaiA's activity, thereby promoting the dephosphorylation phase of the reaction. Therefore, we report here dynamics of interaction of KaiA and KaiB with KaiC that determine the period and amplitude of this in vitro oscillator. PMID:20679240

  18. Crystal Structure of the Redox-Active Cofactor DBMIB Bound to Circadian Clock Protein KaiA and Structural Basis for DBMIB’s Ability to Prevent Stimulation of KaiC Phosphorylation by KaiA

    PubMed Central

    Pattanayek, Rekha; Sidiqi, Said K.; Egli, Martin

    2012-01-01

    KaiA protein that stimulates KaiC phosphorylation in the cyanobacterial circadian clock was recently shown to be destabilized by dibromothymoquinone (DBMIB), thus revealing KaiA as a sensor of the plastoquinone (PQ) redox state and suggesting an indirect control of the clock by light through PQ redox changes. Here we show using X-ray crystallography that several DBMIBs are bound to KaiA dimer. Some binding modes are consistent with oligomerization of N-terminal KaiA pseudoreceiver domains and/or reduced inter-domain flexibility. DBMIB bound to the C-terminal KaiA (C-KaiA) domain and limited stimulation of KaiC kinase activity by C-KaiA in the presence of DBMIB demonstrate that the cofactor may weakly inhibit KaiA-KaiC binding. PMID:23020633

  19. Active output state of the Synechococcus Kai circadian oscillator

    PubMed Central

    Paddock, Mark L.; Boyd, Joseph S.; Adin, Dawn M.; Golden, Susan S.

    2013-01-01

    The mechanisms by which cellular oscillators keep time and transmit temporal information are poorly understood. In cyanobacteria, the timekeeping aspect of the circadian oscillator, composed of the KaiA, KaiB, and KaiC proteins, involves a cyclic progression of phosphorylation states at Ser431 and Thr432 of KaiC. Elucidating the mechanism that uses this temporal information to modulate gene expression is complicated by unknowns regarding the number, structure, and regulatory effects of output components. To identify oscillator signaling states without a complete description of the output machinery, we defined a simple metric, Kai-complex output activity (KOA), that represents the difference in expression of reporter genes between strains that carry specific variants of KaiC and baseline strains that lack KaiC. In the absence of the oscillator, expression of the class 1 paradigm promoter PkaiBC was locked at its usual peak level; conversely, that of the class 2 paradigm promoter PpurF was locked at its trough level. However, for both classes of promoters, peak KOA in wild-type strains coincided late in the circadian cycle near subjective dawn, when KaiC-pST becomes most prevalent (Ser431 is phosphorylated and Thr432 is not). Analogously, peak KOA was detected specifically for the phosphomimetic of KaiC-pST (KaiC-ET). Notably, peak KOA required KaiB, indicating that a KaiBC complex is involved in the output activity. We also found evidence that phosphorylated RpaA (regulator of phycobilisome associated) represses an RpaA-independent output of KOA. A simple mathematical expression successfully simulated two key features of the oscillator—the time of peak KOA and the peak-to-trough amplitude changes. PMID:24043774

  20. The Legionella pneumophila kai operon is implicated in stress response and confers fitness in competitive environments

    PubMed Central

    Loza-Correa, Maria; Sahr, Tobias; Rolando, Monica; Daniels, Craig; Petit, Pierre; Skarina, Tania; Valero, Laura Gomez; Dervins-Ravault, Delphine; Honoré, Nadine; Savchenko, Aleksey; Buchrieser, Carmen

    2014-01-01

    Summary Legionella pneumophila uses aquatic protozoa as replication niche and protection from harsh environments. Although L. pneumophila is not known to have a circadian clock, it encodes homologues of the KaiBC proteins of Cyanobacteria that regulate circadian gene expression. We show that L. pneumophila kaiB, kaiC and the downstream gene lpp1114, are transcribed as a unit under the control of the stress sigma factor RpoS. KaiC and KaiB of L. pneumophila do not interact as evidenced by yeast and bacterial two-hybrid analyses. Fusion of the C-terminal residues of cyanobacterial KaiB to Legionella KaiB restores their interaction. In contrast, KaiC of L. pneumophila conserved autophosphorylation activity, but KaiB does not trigger the dephosphorylation of KaiC like in Cyanobacteria. The crystal structure of L. pneumophila KaiB suggests that it is an oxidoreductase-like protein with a typical thioredoxin fold. Indeed, mutant analyses revealed that the kai operon-encoded proteins increase fitness of L. pneumophila in competitive environments, and confer higher resistance to oxidative and sodium stress. The phylogenetic analysis indicates that L. pneumophila KaiBC resemble Synechosystis KaiC2B2 and not circadian KaiB1C1. Thus, the L. pneumophila Kai proteins do not encode a circadian clock, but enhance stress resistance and adaption to changes in the environments. PMID:23957615

  1. Quantifying the Rhythm of KaiB-C Interaction for In Vitro Cyanobacterial Circadian Clock

    PubMed Central

    Ma, Lan; Ranganathan, Rama

    2012-01-01

    An oscillator consisting of KaiA, KaiB, and KaiC proteins comprises the core of cyanobacterial circadian clock. While one key reaction in this process—KaiC phosphorylation—has been extensively investigated and modeled, other key processes, such as the interactions among Kai proteins, are not understood well. Specifically, different experimental techniques have yielded inconsistent views about Kai A, B, and C interactions. Here, we first propose a mathematical model of cyanobacterial circadian clock that explains the recently observed dynamics of the four phospho-states of KaiC as well as the interactions among the three Kai proteins. Simulations of the model show that the interaction between KaiB and KaiC oscillates with the same period as the phosphorylation of KaiC, but displays a phase delay of ∼8 hr relative to the total phosphorylated KaiC. Secondly, this prediction on KaiB-C interaction are evaluated using a novel FRET (Fluorescence Resonance Energy Transfer)-based assay by tagging fluorescent proteins Cerulean and Venus to KaiC and KaiB, respectively, and reconstituting fluorescent protein-labeled in vitro clock. The data show that the KaiB∶KaiC interaction indeed oscillates with ∼24 hr periodicity and ∼8 hr phase delay relative to KaiC phosphorylation, consistent with model prediction. Moreover, it is noteworthy that our model indicates that the interlinked positive and negative feedback loops are the underlying mechanism for oscillation, with the serine phosphorylated-state (the “S-state") of KaiC being a hub for the feedback loops. Because the kinetics of the KaiB-C interaction faithfully follows that of the S-state, the FRET measurement may provide an important real-time probe in quantitative study of the cyanobacterial circadian clock. PMID:22900029

  2. Dual KaiC-based oscillations constitute the circadian system of cyanobacteria

    PubMed Central

    Kitayama, Yohko; Nishiwaki, Taeko; Terauchi, Kazuki; Kondo, Takao

    2008-01-01

    In the cyanobacterium Synechococcus elongatus PCC 7942, the KaiA, KaiB, and KaiC proteins are essential for the generation of circadian rhythms. Both in vivo and in vitro, phosphorylation of KaiC is regulated positively by KaiA and negatively by KaiB and shows circadian rhythmicity. The autonomous circadian cycling of KaiC phosphorylation is thought to be the basic pacemaker of the circadian clock and to control genome-wide gene expression in cyanobacteria. In this study, we found that temperature-compensated circadian oscillations of gene expression persisted even when KaiC was arrested in the phosphorylated state due to kaiA overexpression. Moreover, two phosphorylation mutants showed transcriptional oscillation with a long period. In kaiA-overexpressing and phosphorylation-deficient strains, KaiC oscillated and transient overexpression of phosphorylation-deficient kaiC reset the phase of the rhythm. These results suggest that transcription- and translation-based oscillations in KaiC abundance are also important for circadian rhythm generation in cyanobacteria. Furthermore, at low temperature, cyanobacteria can show circadian rhythms only when both the KaiC phosphorylation cycle and the transcription and translation cycle are intact. Our findings indicate that multiple coupled oscillatory systems based on the biochemical properties of KaiC are important to maintain robust and precise circadian rhythms in cyanobacteria. PMID:18477603

  3. Intramolecular Regulation of Phosphorylation Status of the Circadian Clock Protein KaiC

    PubMed Central

    Xu, Yao; Mori, Tetsuya; Qin, Ximing; Yan, Heping; Egli, Martin; Johnson, Carl Hirschie

    2009-01-01

    Background KaiC, a central clock protein in cyanobacteria, undergoes circadian oscillations between hypophosphorylated and hyperphosphorylated forms in vivo and in vitro. Structural analyses of KaiC crystals have identified threonine and serine residues in KaiC at three residues (T426, S431, and T432) as potential sites at which KaiC is phosphorylated; mutation of any of these three sites to alanine abolishes rhythmicity, revealing an essential clock role for each residue separately and for KaiC phosphorylation in general. Mass spectrometry studies confirmed that the S431 and T432 residues are key phosphorylation sites, however, the role of the threonine residue at position 426 was not clear from the mass spectrometry measurements. Methodology and Principal Findings Mutational approaches and biochemical analyses of KaiC support a key role for T426 in control of the KaiC phosphorylation status in vivo and in vitro and demonstrates that alternative amino acids at residue 426 dramatically affect KaiC's properties in vivo and in vitro, especially genetic dominance/recessive relationships, KaiC dephosphorylation, and the formation of complexes of KaiC with KaiA and KaiB. These mutations alter key circadian properties, including period, amplitude, robustness, and temperature compensation. Crystallographic analyses indicate that the T426 site is phosphorylatible under some conditions, and in vitro phosphorylation assays of KaiC demonstrate labile phosphorylation of KaiC when the primary S431 and T432 sites are blocked. Conclusions and Significance T426 is a crucial site that regulates KaiC phosphorylation status in vivo and in vitro and these studies underscore the importance of KaiC phosphorylation status in the essential cyanobacterial circadian functions. The regulatory roles of these phosphorylation sites–including T426–within KaiC enhance our understanding of the molecular mechanism underlying circadian rhythm generation in cyanobacteria. PMID:19946629

  4. 78 FR 70096 - Requested Administrative Waiver of the Coastwise Trade Laws: Vessel KOKUA KAI; Invitation for...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-11-22

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF TRANSPORTATION Maritime Administration Requested Administrative Waiver of the Coastwise Trade Laws: Vessel KOKUA KAI... of the vessel KOKUA KAI is: Intended Commercial Use Of Vessel: ``Tour Boat.'' Geographic...

  5. ATPase activity and its temperature compensation of the cyanobacterial clock protein KaiC.

    PubMed

    Murakami, Reiko; Miyake, Ayumi; Iwase, Ryo; Hayashi, Fumio; Uzumaki, Tatsuya; Ishiura, Masahiro

    2008-04-01

    KaiA, KaiB and KaiC constitute the circadian clock machinery in cyanobacteria. KaiC is a homohexamer; its subunit contains duplicated halves, each with a set of ATPase motifs. Here, using highly purified KaiC preparations of the thermophilic cyanobacterium Thermosynechococcus elongatus BP-1 produced in Escherichia coli, we found that the N- and C-terminal domains of KaiC had extremely weak ATPase activity. ATPase activity showed temperature compensation in wild-type KaiC, but not in KaiC(S431A/T432A), a mutant that lacks two phosphorylation sites. We concluded that KaiC phosphorylation is involved in the ATPase temperature-compensation mechanism-which is probably critical to the stability of the circadian clock in cyanobacteria-and we hypothesized the following temperature-compensation mechanism: (i) The C-terminal phosphorylation sites of a KaiC hexamer subunit are phosphorylated by the C-terminal domain of an adjacent KaiC subunit; (ii) the phosphorylation suppresses the ATPase activity of the C-terminal domain; and (iii) the phosphorylated KaiC spontaneously dephosphorylates, resulting in the recover of ATPase activity. PMID:18363969

  6. Circadian autodephosphorylation of cyanobacterial clock protein KaiC occurs via formation of ATP as intermediate.

    PubMed

    Nishiwaki, Taeko; Kondo, Takao

    2012-05-25

    The cyanobacterial circadian oscillator can be reconstituted in vitro; mixing three clock proteins (KaiA, KaiB, and KaiC) with ATP results in an oscillation of KaiC phosphorylation with a periodicity of ~24 h. The hexameric ATPase KaiC hydrolyzes ATP bound at subunit interfaces. KaiC also exhibits autokinase and autophosphatase activities, the latter of which is particularly noteworthy because KaiC is phylogenetically distinct from typical protein phosphatases. To examine this activity, we performed autodephosphorylation assays using (32)P-labeled KaiC. The residual radioactive ATP bound to subunit interfaces was removed using a newly established method, which included the dissociation of KaiC hexamers into monomers and the reconstitution of KaiC hexamers with nonradioactive ATP. This approach ensured that only the signals derived from (32)P-labeled KaiC were examined. We detected the transient formation of [(32)P]ATP preceding the accumulation of (32)P(i). Together with kinetic analyses, our data demonstrate that KaiC undergoes dephosphorylation via a mechanism that differs from those of conventional protein phosphatases. A phosphate group at a phosphorylation site is first transferred to KaiC-bound ADP to form ATP as an intermediate, which can be regarded as a reversal of the autophosphorylation reaction. Subsequently, the ATP molecule is hydrolyzed to form P(i). We propose that the ATPase active site mediates not only ATP hydrolysis but also the bidirectional transfer of the phosphate between phosphorylation sites and the KaiC-bound nucleotide. On the basis of these findings, we can now dissect the dynamics of the KaiC phosphorylation cycle relative to ATPase activity. PMID:22493509

  7. Circadian Autodephosphorylation of Cyanobacterial Clock Protein KaiC Occurs via Formation of ATP as Intermediate*

    PubMed Central

    Nishiwaki, Taeko; Kondo, Takao

    2012-01-01

    The cyanobacterial circadian oscillator can be reconstituted in vitro; mixing three clock proteins (KaiA, KaiB, and KaiC) with ATP results in an oscillation of KaiC phosphorylation with a periodicity of ∼24 h. The hexameric ATPase KaiC hydrolyzes ATP bound at subunit interfaces. KaiC also exhibits autokinase and autophosphatase activities, the latter of which is particularly noteworthy because KaiC is phylogenetically distinct from typical protein phosphatases. To examine this activity, we performed autodephosphorylation assays using 32P-labeled KaiC. The residual radioactive ATP bound to subunit interfaces was removed using a newly established method, which included the dissociation of KaiC hexamers into monomers and the reconstitution of KaiC hexamers with nonradioactive ATP. This approach ensured that only the signals derived from 32P-labeled KaiC were examined. We detected the transient formation of [32P]ATP preceding the accumulation of 32Pi. Together with kinetic analyses, our data demonstrate that KaiC undergoes dephosphorylation via a mechanism that differs from those of conventional protein phosphatases. A phosphate group at a phosphorylation site is first transferred to KaiC-bound ADP to form ATP as an intermediate, which can be regarded as a reversal of the autophosphorylation reaction. Subsequently, the ATP molecule is hydrolyzed to form Pi. We propose that the ATPase active site mediates not only ATP hydrolysis but also the bidirectional transfer of the phosphate between phosphorylation sites and the KaiC-bound nucleotide. On the basis of these findings, we can now dissect the dynamics of the KaiC phosphorylation cycle relative to ATPase activity. PMID:22493509

  8. Conversion between two conformational states of KaiC is induced by ATP hydrolysis as a trigger for cyanobacterial circadian oscillation.

    PubMed

    Oyama, Katsuaki; Azai, Chihiro; Nakamura, Kaori; Tanaka, Syun; Terauchi, Kazuki

    2016-01-01

    The cyanobacterial circadian oscillator can be reconstituted in vitro by mixing three clock proteins, KaiA, KaiB and KaiC, with ATP. KaiC is the only protein with circadian rhythmic activities. In the present study, we tracked the complex formation of the three Kai proteins over time using blue native (BN) polyacrylamide gel electrophoresis (PAGE), in which proteins are charged with the anionic dye Coomassie brilliant blue (CBB). KaiC was separated as three bands: the KaiABC complex, KaiC hexamer and KaiC monomer. However, no KaiC monomer was observed using gel filtration chromatography and CBB-free native PAGE. These data indicate two conformational states of KaiC hexamer and show that the ground-state KaiC (gs-KaiC) is stable and competent-state KaiC (cs-KaiC) is labile and degraded into monomers by the binding of CBB. Repeated conversions from gs-KaiC to cs-KaiC were observed over 24 h using an in vitro reconstitution system. Phosphorylation of KaiC promoted the conversion from gs-KaiC to cs-KaiC. KaiA sustained the gs-KaiC state, and KaiB bound only cs-KaiC. An E77Q/E78Q-KaiC variant that lacked N-terminal ATPase activity remained in the gs-KaiC state. Taken together, ATP hydrolysis induces the formation of cs-KaiC and promotes the binding of KaiB, which is a trigger for circadian oscillations. PMID:27580682

  9. Conversion between two conformational states of KaiC is induced by ATP hydrolysis as a trigger for cyanobacterial circadian oscillation

    PubMed Central

    Oyama, Katsuaki; Azai, Chihiro; Nakamura, Kaori; Tanaka, Syun; Terauchi, Kazuki

    2016-01-01

    The cyanobacterial circadian oscillator can be reconstituted in vitro by mixing three clock proteins, KaiA, KaiB and KaiC, with ATP. KaiC is the only protein with circadian rhythmic activities. In the present study, we tracked the complex formation of the three Kai proteins over time using blue native (BN) polyacrylamide gel electrophoresis (PAGE), in which proteins are charged with the anionic dye Coomassie brilliant blue (CBB). KaiC was separated as three bands: the KaiABC complex, KaiC hexamer and KaiC monomer. However, no KaiC monomer was observed using gel filtration chromatography and CBB-free native PAGE. These data indicate two conformational states of KaiC hexamer and show that the ground-state KaiC (gs-KaiC) is stable and competent-state KaiC (cs-KaiC) is labile and degraded into monomers by the binding of CBB. Repeated conversions from gs-KaiC to cs-KaiC were observed over 24 h using an in vitro reconstitution system. Phosphorylation of KaiC promoted the conversion from gs-KaiC to cs-KaiC. KaiA sustained the gs-KaiC state, and KaiB bound only cs-KaiC. An E77Q/E78Q-KaiC variant that lacked N-terminal ATPase activity remained in the gs-KaiC state. Taken together, ATP hydrolysis induces the formation of cs-KaiC and promotes the binding of KaiB, which is a trigger for circadian oscillations. PMID:27580682

  10. Fluorescence Correlation Spectroscopy to Monitor Kai Protein-based Circadian Oscillations in Real Time*

    PubMed Central

    Goda, Kazuhito; Ito, Hiroshi; Kondo, Takao; Oyama, Tokitaka

    2012-01-01

    Dynamic protein-protein interactions play an essential role in cellular regulatory systems. The cyanobacterial circadian clock is an oscillatory system that can be reconstituted in vitro by mixing ATP and three clock proteins: KaiA, KaiB, and KaiC. Association and dissociation of KaiB from KaiC-containing complexes are critical to circadian phosphorylation and dephosphorylation of KaiC. We developed an automated and noninvasive method to monitor dynamic complex formation in real time using confocal fluorescence correlation spectroscopy (FCS) and uniformly labeled KaiB as a probe. A nanomolar concentration of the labeled KaiB for FCS measurement did not interfere with the oscillatory system but behaved similarly to the wild-type one during the measurement period (>5 days). The fluorescent probe was stable against repeated laser exposure. As an application, we show that this detection system allowed analysis of the dynamics of both long term circadian oscillations and short term responses to temperature changes (∼10 min) in the same sample. This suggested that a phase shift of the clock with a high temperature pulse occurred just after the stimulus through dissociation of KaiB from the KaiC complex. This monitoring method should improve our understanding of the mechanisms underlying this cellular circadian oscillator and provide a means to assess dynamic protein interactions in biological systems characterized by rates similar to those observed with the Kai proteins. PMID:22157012

  11. Structural basis of unique ligand specificity of KAI2-like protein from parasitic weed Striga hermonthica

    PubMed Central

    Xu, Yuqun; Miyakawa, Takuya; Nakamura, Hidemitsu; Nakamura, Akira; Imamura, Yusaku; Asami, Tadao; Tanokura, Masaru

    2016-01-01

    The perception of two plant germination inducers, karrikins and strigolactones, are mediated by the proteins KAI2 and D14. Recently, KAI2-type proteins from parasitic weeds, which are possibly related to seed germination induced by strigolactone, have been classified into three clades characterized by different responses to karrikin/strigolactone. Here we characterized a karrikin-binding protein in Striga (ShKAI2iB) that belongs to intermediate-evolving KAI2 and provided the structural bases for its karrikin-binding specificity. Binding assays showed that ShKAI2iB bound karrikins but not strigolactone, differing from other KAI2 and D14. The crystal structures of ShKAI2iB and ShKAI2iB-karrikin complex revealed obvious structural differences in a helix located at the entry of its ligand-binding cavity. This results in a smaller closed pocket, which is also the major cause of ShKAI2iB’s specificity of binding karrikin. Our structural study also revealed that a few non-conserved amino acids led to the distinct ligand-binding profile of ShKAI2iB, suggesting that the evolution of KAI2 resulted in its diverse functions. PMID:27507097

  12. Structural basis of unique ligand specificity of KAI2-like protein from parasitic weed Striga hermonthica.

    PubMed

    Xu, Yuqun; Miyakawa, Takuya; Nakamura, Hidemitsu; Nakamura, Akira; Imamura, Yusaku; Asami, Tadao; Tanokura, Masaru

    2016-01-01

    The perception of two plant germination inducers, karrikins and strigolactones, are mediated by the proteins KAI2 and D14. Recently, KAI2-type proteins from parasitic weeds, which are possibly related to seed germination induced by strigolactone, have been classified into three clades characterized by different responses to karrikin/strigolactone. Here we characterized a karrikin-binding protein in Striga (ShKAI2iB) that belongs to intermediate-evolving KAI2 and provided the structural bases for its karrikin-binding specificity. Binding assays showed that ShKAI2iB bound karrikins but not strigolactone, differing from other KAI2 and D14. The crystal structures of ShKAI2iB and ShKAI2iB-karrikin complex revealed obvious structural differences in a helix located at the entry of its ligand-binding cavity. This results in a smaller closed pocket, which is also the major cause of ShKAI2iB's specificity of binding karrikin. Our structural study also revealed that a few non-conserved amino acids led to the distinct ligand-binding profile of ShKAI2iB, suggesting that the evolution of KAI2 resulted in its diverse functions. PMID:27507097

  13. A protocol for preparing nucleotide-free KaiC monomer.

    PubMed

    Mukaiyama, Atsushi; Osako, Masato; Hikima, Takaaki; Kondo, Takao; Akiyama, Shuji

    2015-01-01

    The hexameric form of the KaiC protein is a core of the cyanobacterial biological clock, and its enzymatic activities exhibit circadian periodicity. The instability of the monomeric form of nucleotide-free KaiC has precluded its storage and detailed analyses of the activities of the reassembled hexamer. Here, we provide a protocol for preparing nucleotide-free KaiC monomer that is stable in solution and for triggering its reassembly into intact KaiC hexamer by the addition of ATP. A phosphate buffer containing glutamic acid and arginine enhanced the stability of KaiC monomer considerably. In addition, we found that reassembled KaiC hexamer was functionally active as the intact hexamer. This protocol provides a methodological basis for further analyses of first-turnover events of the ATPase/autokinase/autophosphatase activities of the KaiC hexamer. PMID:27493519

  14. A protocol for preparing nucleotide-free KaiC monomer

    PubMed Central

    Mukaiyama, Atsushi; Osako, Masato; Hikima, Takaaki; Kondo, Takao; Akiyama, Shuji

    2015-01-01

    The hexameric form of the KaiC protein is a core of the cyanobacterial biological clock, and its enzymatic activities exhibit circadian periodicity. The instability of the monomeric form of nucleotide-free KaiC has precluded its storage and detailed analyses of the activities of the reassembled hexamer. Here, we provide a protocol for preparing nucleotide-free KaiC monomer that is stable in solution and for triggering its reassembly into intact KaiC hexamer by the addition of ATP. A phosphate buffer containing glutamic acid and arginine enhanced the stability of KaiC monomer considerably. In addition, we found that reassembled KaiC hexamer was functionally active as the intact hexamer. This protocol provides a methodological basis for further analyses of first-turnover events of the ATPase/autokinase/autophosphatase activities of the KaiC hexamer. PMID:27493519

  15. SITE TECHNOLOGY CAPSULE: KAI RADIO FREQUENCY HEATING TECHNOLOGY

    EPA Science Inventory

    KAI developed a patented, in situ RFH technology to enhance the removal of volatile and semi-volatile organics by soil vapor extraction (SVE). Electromagnetic energy heats the soil resulting in increased contaminant vapor pressures and soil permeability that may increase with dry...

  16. INNOVATIVE TECHNOLOGY EVALUATION REPORT: RADIO FREQUENCY HEATING, KAI TECHNOLOGIES, INC.

    EPA Science Inventory

    A demonstration of KAI Technologies in-situ radio frequency heating system for soil treatment was conducted from January 1994 to July 1994 at Kelly Air Force Base in San Antonio, Texas. This demonstration was conducted as a joint effort between the USEPA and the USAF. The technol...

  17. DEMONSTRATION BULLETIN: RADIO FREQUENCY HEATING - KAI TECHNOLOGIES, INC.

    EPA Science Inventory

    Radio frequency heating (RFH) is a process that uses electromagnetic energy in the radio frequency (RF) band to heat soil in situ, thereby potentially enhancing the performance of standard soil vapor extraction (SVE) technologies. An RFH system developed by KAI Technologies, I...

  18. Exchange of ADP with ATP in the CII ATPase domain promotes autophosphorylation of cyanobacterial clock protein KaiC

    PubMed Central

    Nishiwaki-Ohkawa, Taeko; Kitayama, Yohko; Ochiai, Erika; Kondo, Takao

    2014-01-01

    The cyanobacterial circadian oscillator can be reconstituted in vitro. In the presence of KaiA and KaiB, the phosphorylation state of KaiC oscillates with a periodicity of ∼24 h. KaiC is a hexameric P-loop ATPase with autophosphorylation and autodephosphorylation activities. Recently, we found that dephosphorylation of KaiC occurs via reversal of the phosphorylation reaction: a phosphate group attached to Ser431/Thr432 is transferred to KaiC-bound ADP to generate ATP, which is subsequently hydrolyzed. This unusual reaction mechanism suggests that the KaiC phosphorylation rhythm is sustained by periodic shifts in the equilibrium of the reversible autophosphorylation reaction, the molecular basis of which has never been elucidated. Because KaiC-bound ATP and ADP serve as substrates for the forward and reverse reactions, respectively, we investigated the regulation of the nucleotide-bound state of KaiC. In the absence of KaiA, the condition in which the reverse reaction proceeds, KaiC favored the ADP-bound state. KaiA increased the ratio of ATP to total KaiC-bound nucleotides by facilitating the release of bound ADP and the incorporation of exogenous ATP, allowing the forward reaction to proceed. When both KaiA and KaiB were present, the ratio of ATP to total bound nucleotides exhibited a circadian rhythm, whose phase was advanced by several hours relative to that of the phosphorylation rhythm. Based on these findings, we propose that the direction of the reversible autophosphorylation reaction is regulated by KaiA and KaiB at the level of substrate availability and that this regulation sustains the oscillation of the phosphorylation state of KaiC. PMID:24616498

  19. Exchange of ADP with ATP in the CII ATPase domain promotes autophosphorylation of cyanobacterial clock protein KaiC.

    PubMed

    Nishiwaki-Ohkawa, Taeko; Kitayama, Yohko; Ochiai, Erika; Kondo, Takao

    2014-03-25

    The cyanobacterial circadian oscillator can be reconstituted in vitro. In the presence of KaiA and KaiB, the phosphorylation state of KaiC oscillates with a periodicity of ∼24 h. KaiC is a hexameric P-loop ATPase with autophosphorylation and autodephosphorylation activities. Recently, we found that dephosphorylation of KaiC occurs via reversal of the phosphorylation reaction: a phosphate group attached to Ser431/Thr432 is transferred to KaiC-bound ADP to generate ATP, which is subsequently hydrolyzed. This unusual reaction mechanism suggests that the KaiC phosphorylation rhythm is sustained by periodic shifts in the equilibrium of the reversible autophosphorylation reaction, the molecular basis of which has never been elucidated. Because KaiC-bound ATP and ADP serve as substrates for the forward and reverse reactions, respectively, we investigated the regulation of the nucleotide-bound state of KaiC. In the absence of KaiA, the condition in which the reverse reaction proceeds, KaiC favored the ADP-bound state. KaiA increased the ratio of ATP to total KaiC-bound nucleotides by facilitating the release of bound ADP and the incorporation of exogenous ATP, allowing the forward reaction to proceed. When both KaiA and KaiB were present, the ratio of ATP to total bound nucleotides exhibited a circadian rhythm, whose phase was advanced by several hours relative to that of the phosphorylation rhythm. Based on these findings, we propose that the direction of the reversible autophosphorylation reaction is regulated by KaiA and KaiB at the level of substrate availability and that this regulation sustains the oscillation of the phosphorylation state of KaiC. PMID:24616498

  20. KaiC intersubunit communication facilitates robustness of circadian rhythms in cyanobacteria.

    PubMed

    Kitayama, Yohko; Nishiwaki-Ohkawa, Taeko; Sugisawa, Yukiko; Kondo, Takao

    2013-01-01

    The cyanobacterial circadian clock is the only model clock to have been reconstituted in vitro. KaiC, the central clock component, is a homohexameric ATPase with autokinase and autophosphatase activities. Changes in phosphorylation state have been proposed to switch KaiC's activity between autokinase and autophosphatase. Here we analyse the molecular mechanism underlying the regulation of KaiC's activity, in the context of its hexameric structure. We reconstitute KaiC hexamers containing different variant protomers, and measure their autophosphatase and autokinase activities. We identify two types of regulatory mechanisms with distinct functions. First, local interactions between adjacent phosphorylation sites regulate KaiC's activities, coupling the ATPase and nucleotide-binding states at subunit interfaces of the CII domain. Second, the phosphorylation states of the protomers affect the overall activity of KaiC hexamers via intersubunit communication. Our findings indicate that intra-hexameric interactions play an important role in sustaining robust circadian rhythmicity. PMID:24305644

  1. Roles of two ATPase-motif-containing domains in cyanobacterial circadian clock protein KaiC.

    PubMed

    Hayashi, Fumio; Itoh, Noriyo; Uzumaki, Tatsuya; Iwase, Ryo; Tsuchiya, Yuka; Yamakawa, Hisanori; Morishita, Megumi; Onai, Kiyoshi; Itoh, Shigeru; Ishiura, Masahiro

    2004-12-10

    Cyanobacterial clock protein KaiC has a hexagonal, pot-shaped structure composed of six identical dumbbell-shaped subunits. Each subunit has duplicated domains, and each domain has a set of ATPase motifs. The two spherical regions of the dumbbell are likely to correspond to two domains. We examined the role of the two sets of ATPase motifs by analyzing the in vitro activity of ATPgammaS binding, AMPPNP-induced hexamerization, thermostability, and phosphorylation of KaiC and by in vivo rhythm assays both in wild type KaiC (KaiCWT) and KaiCs carrying mutations in either Walker motif A or deduced catalytic Glu residues. We demonstrated that 1) the KaiC subunit had two types of ATP-binding sites, a high affinity site in N-terminal ATPase motifs and a low affinity site in C-terminal ATPase motifs, 2) the N-terminal motifs were responsible for hexamerization, and 3) the C-terminal motifs were responsible for both stabilization and phosphorylation of the KaiC hexamer. We proposed the following reaction mechanism. ATP preferentially binds to the N-terminal high affinity site, inducing the hexamerization of KaiC. Additional ATP then binds to the C-terminal low affinity site, stabilizing and phosphorylating the hexamer. We discussed the effect of these KaiC mutations on circadian bioluminescence rhythm in cells of cyanobacteria. PMID:15377674

  2. Dephosphorylation of the Core Clock Protein KaiC in the Cyanobacterial KaiABC Circadian Oscillator Proceeds via an ATP Synthase Mechanism

    SciTech Connect

    Egli, Martin; Mori, Tetsuya; Pattanayek, Rekha; Xu, Yao; Qin, Ximing; Johnson, Carl H.

    2014-10-02

    The circadian clock of the cyanobacterium Synechococcus elongatus can be reconstituted in vitro from three proteins, KaiA, KaiB, and KaiC in the presence of ATP, to tick in a temperature-compensated manner. KaiC, the central cog of this oscillator, forms a homohexamer with 12 ATP molecules bound between its N- and C-terminal domains and exhibits unusual properties. Both the N-terminal (CI) and C-terminal (CII) domains harbor ATPase activity, and the subunit interfaces between CII domains are the sites of autokinase and autophosphatase activities. Hydrolysis of ATP correlates with phosphorylation at threonine and serine sites across subunits in an orchestrated manner, such that first T432 and then S431 are phosphorylated, followed by dephosphorylation of these residues in the same order. Although structural work has provided insight into the mechanisms of ATPase and kinase, the location and mechanism of the phosphatase have remained enigmatic. From the available experimental data based on a range of approaches, including KaiC crystal structures and small-angle X-ray scattering models, metal ion dependence, site-directed mutagenesis (i.e., E318, the general base), and measurements of the associated clock periods, phosphorylation patterns, and dephosphorylation courses as well as a lack of sequence motifs in KaiC that are typically associated with known phosphatases, we hypothesized that KaiCII makes use of the same active site for phosphorylation and dephosphorlyation. We observed that wild-type KaiC (wt-KaiC) exhibits an ATP synthase activity that is significantly reduced in the T432A/S431A mutant. We interpret the first observation as evidence that KaiCII is a phosphotransferase instead of a phosphatase and the second that the enzyme is capable of generating ATP, both from ADP and P{sub i} (in a reversal of the ATPase reaction) and from ADP and P-T432/P-S431 (dephosphorylation). This new concept regarding the mechanism of dephosphorylation is also supported by the

  3. Dephosphorylation of the core clock protein KaiC in the cyanobacterial KaiABC circadian oscillator proceeds via an ATP synthase mechanism.

    PubMed

    Egli, Martin; Mori, Tetsuya; Pattanayek, Rekha; Xu, Yao; Qin, Ximing; Johnson, Carl H

    2012-02-28

    The circadian clock of the cyanobacterium Synechococcus elongatus can be reconstituted in vitro from three proteins, KaiA, KaiB, and KaiC in the presence of ATP, to tick in a temperature-compensated manner. KaiC, the central cog of this oscillator, forms a homohexamer with 12 ATP molecules bound between its N- and C-terminal domains and exhibits unusual properties. Both the N-terminal (CI) and C-terminal (CII) domains harbor ATPase activity, and the subunit interfaces between CII domains are the sites of autokinase and autophosphatase activities. Hydrolysis of ATP correlates with phosphorylation at threonine and serine sites across subunits in an orchestrated manner, such that first T432 and then S431 are phosphorylated, followed by dephosphorylation of these residues in the same order. Although structural work has provided insight into the mechanisms of ATPase and kinase, the location and mechanism of the phosphatase have remained enigmatic. From the available experimental data based on a range of approaches, including KaiC crystal structures and small-angle X-ray scattering models, metal ion dependence, site-directed mutagenesis (i.e., E318, the general base), and measurements of the associated clock periods, phosphorylation patterns, and dephosphorylation courses as well as a lack of sequence motifs in KaiC that are typically associated with known phosphatases, we hypothesized that KaiCII makes use of the same active site for phosphorylation and dephosphorlyation. We observed that wild-type KaiC (wt-KaiC) exhibits an ATP synthase activity that is significantly reduced in the T432A/S431A mutant. We interpret the first observation as evidence that KaiCII is a phosphotransferase instead of a phosphatase and the second that the enzyme is capable of generating ATP, both from ADP and P(i) (in a reversal of the ATPase reaction) and from ADP and P-T432/P-S431 (dephosphorylation). This new concept regarding the mechanism of dephosphorylation is also supported by the

  4. ATPase activity of KaiC determines the basic timing for circadian clock of cyanobacteria.

    PubMed

    Terauchi, Kazuki; Kitayama, Yohko; Nishiwaki, Taeko; Miwa, Kumiko; Murayama, Yoriko; Oyama, Tokitaka; Kondo, Takao

    2007-10-01

    Self-sustainable oscillation of KaiC phosphorylation has been reconstituted in vitro, demonstrating that this cycle is the basic time generator of the circadian clock of cyanobacteria. Here we show that the ATPase activity of KaiC satisfies the characteristics of the circadian oscillation, the period length, and the temperature compensation. KaiC possesses extremely weak but stable ATPase activity (15 molecules of ATP per day), and the addition of KaiA and KaiB makes the activity oscillate with a circadian period in vitro. The ATPase activity of KaiC is inherently temperature-invariant, suggesting that temperature compensation of the circadian period could be driven by this simple biochemical reaction. Moreover, the activities of wild-type KaiC and five period-mutant proteins are directly proportional to their in vivo circadian frequencies, indicating that the ATPase activity defines the circadian period. Thus, we propose that KaiC ATPase activity constitutes the most fundamental reaction underlying circadian periodicity in cyanobacteria. PMID:17901204

  5. 78 FR 11270 - Requested Administrative Waiver of the Coastwise Trade Laws: Vessel KAI ORA; Invitation for...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-02-15

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF TRANSPORTATION Maritime Administration Requested Administrative Waiver of the Coastwise Trade Laws: Vessel KAI ORA... intended service of the vessel KAI ORA is: Intended Commercial Use of Vessel: ``Skippered Charter in...

  6. 76 FR 67251 - Requested Administrative Waiver of the Coastwise Trade Laws: Vessel KAUHALE KAI III; Invitation...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-10-31

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF TRANSPORTATION Maritime Administration Requested Administrative Waiver of the Coastwise Trade Laws: Vessel KAUHALE KAI III... the vessel KAUHALE KAI III is: INTENDED COMMERCIAL USE OF VESSEL: ``Short term private...

  7. Dephosphorylation of the Core Clock Protein KaiC in the Cyanobacterial KaiABC Circadian Oscillator Proceeds via an ATP Synthase Mechanism

    PubMed Central

    Egli, Martin; Mori, Tetsuya; Pattanayek, Rekha; Xu, Yao; Qin, Ximing; Johnson, Carl H.

    2012-01-01

    The circadian clock of the cyanobacterium Synechococcus elongatus can be reconstituted in vitro from three proteins, KaiA, KaiB and KaiC in the presence of ATP, to tick in a temperature-compensated manner. KaiC, the central cog of this oscillator, forms a homo-hexamer with twelve ATP molecules bound between its N- and C-terminal domains and exhibits unusual properties. Both the N-terminal (CI) and C-terminal (CII) domains harbor ATPase activity and the subunit interfaces between CII domains are the sites of auto-kinase and auto-phosphatase activities. Hydrolysis of ATP correlates with phosphorylation at threonine and serine sites across subunits in an orchestrated manner, such that first T432 and then S431 is phosphorylated, followed by dephosphorylation of these residues in the same order. Although structural work has provided insight into the mechanisms of ATPase and kinase, the location and mechanism of the phosphatase have remained enigmatic. From the available experimental data based on a range of approaches, including KaiC crystal structures and small angle X-ray scattering (SAXS) models, metal ion dependence, site-directed mutagenesis (i.e. E318, the general base) and measurements of the associated clock periods, phosphorylation patterns and dephosphorylation courses as well as a lack of sequence motifs in KaiC that are typically associated with known phosphatases, we hypothesized that KaiCII makes use of the same active site for phosphorylation and dephosphorlyation. We observed that wt-KaiC exhibits an ATP synthase activity that is significantly reduced in the T432A/S431A mutant. We interpret the first observation as evidence that KaiCII is a phospho-transferase instead of a phosphatase and the second that the enzyme is capable of generating ATP, both from ADP + Pi (in a reversal of the ATPase reaction), and ADP + P-T432/P-S431 (dephosphorylation). This new concept regarding the mechanism of dephosphorylation is also supported by strikingly similar make

  8. Phase-dependent generation and transmission of time information by the KaiABC circadian clock oscillator through SasA-KaiC interaction in cyanobacteria.

    PubMed

    Valencia S, J; Bitou, Kyouhei; Ishii, Kentaro; Murakami, Reiko; Morishita, Megumi; Onai, Kiyoshi; Furukawa, Yukio; Imada, Katsumi; Namba, Keiichi; Ishiura, Masahiro

    2012-05-01

    Circadian clocks allow organisms to predict environmental changes of the day/night cycle. In the cyanobacterial circadian clock machinery, the phosphorylation level and ATPase activity of the clock protein KaiC oscillate with a period of approximately 24 h. The time information is transmitted from KaiC to the histidine kinase SasA through the SasA autophosphorylation-enhancing activity of KaiC, ultimately resulting in genome-wide transcription cycles. Here, we showed that SasA derived from the thermophilic cyanobacterium Thermosynechococcus elongatus BP-1 has the domain structure of an orthodox histidine kinase and that its C-terminal domain, which contains a phosphorylation site at His160, is responsible for the autophosphorylation activity and the temperature- and phosphorylation state-dependent trimerization / hexamerization activity of SasA. SasA and KaiC associate through their N-terminal domains with an affinity that depends on their phosphorylation states. Furthermore, the SasA autophosphorylation-enhancing activity of KaiC requires the C-terminal ATPase catalytic site and depends on its phosphorylation state. We show that the phosphotransfer activity of SasA is essential for the generation of normal circadian gene expression in cyanobacterial cells. Numerical simulations suggest that circadian time information (free phosphorylated SasA) is released mainly by unphosphorylated KaiC during the late subjective night. PMID:22512339

  9. Innovative technology evaluation report, radio frequency heating, KAI Technologies, Inc.

    SciTech Connect

    Groeber, M.; Krietemeyer, S.; Saylor, E.

    1995-04-01

    A demonstration of KAI Technologies in-situ radio frequency heating system for soil treatment was conducted from January 1994 to July 1994 at Kelly Air Force Base in San Antonio, Texas. This demonstration was conducted as a joint effort between the USEAP and the USAF. The technology was used to remove hydrocarbon contamination from a contaminated site that had been used during waste treatment and storage operations in the past. This report discusses the results of this demonstration and presents an economic analysis of the process.

  10. Tracking and visualizing the circadian ticking of the cyanobacterial clock protein KaiC in solution.

    PubMed

    Murayama, Yoriko; Mukaiyama, Atsushi; Imai, Keiko; Onoue, Yasuhiro; Tsunoda, Akina; Nohara, Atsushi; Ishida, Tatsuro; Maéda, Yuichiro; Terauchi, Kazuki; Kondo, Takao; Akiyama, Shuji

    2011-01-01

    The circadian clock in cyanobacteria persists even without the transcription/translation feedbacks proposed for eukaryotic systems. The period of the cyanobacterial clock is tuned to the circadian range by the ATPase activity of a clock protein known as KaiC. Here, we provide structural evidence on how KaiC ticks away 24 h while coupling the ATPase activity in its N-terminal ring to the phosphorylation state in its C-terminal ring. During the phosphorylation cycle, the C-terminal domains of KaiC are repositioned in a stepwise manner to affect global expansion and contraction motions of the C-terminal ring. Arg393 of KaiC has a critical function in expanding the C-terminal ring and its replacement with Cys affects the temperature compensation of the period--a fundamental property of circadian clocks. The conformational ticking of KaiC observed here in solution serves as a timing cue for assembly/disassembly of other clock proteins (KaiA and KaiB), and is interlocked with its auto-inhibitory ATPase underlying circadian periodicity of cyanobacteria. PMID:21113137

  11. The Roles of the Dimeric and Tetrameric Structures of the Clock Protein KaiB in the Generation of Circadian Oscillations in Cyanobacteria*

    PubMed Central

    Murakami, Reiko; Mutoh, Risa; Iwase, Ryo; Furukawa, Yukio; Imada, Katsumi; Onai, Kiyoshi; Morishita, Megumi; Yasui, So; Ishii, Kentaro; Valencia Swain, Jonathan Orville; Uzumaki, Tatsuya; Namba, Keiichi; Ishiura, Masahiro

    2012-01-01

    The molecular machinery of the cyanobacterial circadian clock consists of three proteins, KaiA, KaiB, and KaiC. The three Kai proteins interact with each other and generate circadian oscillations in vitro in the presence of ATP (an in vitro KaiABC clock system). KaiB consists of four subunits organized as a dimer of dimers, and its overall shape is that of an elongated hexagonal plate with a positively charged cleft flanked by two negatively charged ridges. We found that a mutant KaiB with a C-terminal deletion (KaiB1–94), which lacks the negatively charged ridges, was a dimer. Despite its dimeric structure, KaiB1–94 interacted with KaiC and generated normal circadian oscillations in the in vitro KaiABC clock system. KaiB1–94 also generated circadian oscillations in cyanobacterial cells, but they were weak, indicating that the C-terminal region and tetrameric structure of KaiB are necessary for the generation of normal gene expression rhythms in vivo. KaiB1–94 showed the highest affinity for KaiC among the KaiC-binding proteins we examined and inhibited KaiC from forming a complex with SasA, which is involved in the main output pathway from the KaiABC clock oscillator in transcription regulation. This defect explains the mechanism underlying the lack of normal gene expression rhythms in cells expressing KaiB1–94. PMID:22722936

  12. Evidence that KARRIKIN-INSENSITIVE2 (KAI2) Receptors may Perceive an Unknown Signal that is not Karrikin or Strigolactone

    PubMed Central

    Conn, Caitlin E.; Nelson, David C.

    2016-01-01

    The α/β-hydrolases KAI2 and D14 are paralogous receptors for karrikins and strigolactones, two classes of plant growth regulators with butenolide moieties. KAI2 and D14 act in parallel signaling pathways that share a requirement for the F-box protein MAX2, but produce distinct growth responses by regulating different members of the SMAX1-LIKE/D53 family. kai2 and max2 mutants share seed germination, seedling growth, leaf shape, and petiole orientation phenotypes that are not found in d14 or SL-deficient mutants. This implies that KAI2 recognizes an unknown, endogenous signal, herein termed KAI2 ligand (KL). Recent studies of ligand-specificity among KAI2 paralogs in basal land plants and root parasitic plants suggest that karrikin and strigolactone perception may be evolutionary adaptations of KL receptors. Here we demonstrate that evolutionarily conserved KAI2c genes from two parasite species rescue multiple phenotypes of the Arabidopsis kai2 mutant, unlike karrikin-, and strigolactone-specific KAI2 paralogs. We hypothesize that KAI2c proteins recognize KL, which could be an undiscovered hormone. PMID:26779242

  13. KaiC intersubunit communication facilitates robustness of circadian rhythms in cyanobacteria

    PubMed Central

    Kitayama, Yohko; Nishiwaki-Ohkawa, Taeko; Sugisawa, Yukiko; Kondo, Takao

    2013-01-01

    The cyanobacterial circadian clock is the only model clock to have been reconstituted in vitro. KaiC, the central clock component, is a homohexameric ATPase with autokinase and autophosphatase activities. Changes in phosphorylation state have been proposed to switch KaiC’s activity between autokinase and autophosphatase. Here we analyse the molecular mechanism underlying the regulation of KaiC’s activity, in the context of its hexameric structure. We reconstitute KaiC hexamers containing different variant protomers, and measure their autophosphatase and autokinase activities. We identify two types of regulatory mechanisms with distinct functions. First, local interactions between adjacent phosphorylation sites regulate KaiC’s activities, coupling the ATPase and nucleotide-binding states at subunit interfaces of the CII domain. Second, the phosphorylation states of the protomers affect the overall activity of KaiC hexamers via intersubunit communication. Our findings indicate that intra-hexameric interactions play an important role in sustaining robust circadian rhythmicity. PMID:24305644

  14. An arginine tetrad as mediator of input-dependent and input-independent ATPases in the clock protein KaiC

    PubMed Central

    Pattanayek, Rekha; Xu, Yao; Lamichhane, Aashish; Johnson, Carl H.; Egli, Martin

    2014-01-01

    A post-translational oscillator (PTO) composed of the proteins KaiA, KaiB and KaiC is at the heart of the cyanobacterial circadian clock. KaiC interacts with KaiA and KaiB over the daily cycle, and CII domains undergo rhythmic phosphorylation/dephosphorylation with a 24 h period. Both the N-terminal (CI) and C-terminal (CII) rings of KaiC exhibit ATPase activity. The CI ATPase proceeds in an input-independent fashion, but the CII ATPase is subject to metabolic input signals. The crystal structure of KaiC from Thermosynechococcus elongatus allows insight into the different anatomies of the CI and CII ATPases. Four consecutive arginines in CI (Arg linker) that connect the P-loop, CI subunits and CI and CII at the ring interface are primary candidates for the coordination of the CI and CII activities. The mutation of linker residues alters the period or triggers arhythmic behavior. Comparison between the CI and CII structures also reveals differences in loop regions that are key to KaiA and KaiB binding and activation of CII ATPase and kinase. Common packing features in KaiC crystals shed light on the KaiB–KaiC interaction. PMID:24816106

  15. An arginine tetrad as mediator of input-dependent and input-independent ATPases in the clock protein KaiC.

    PubMed

    Pattanayek, Rekha; Xu, Yao; Lamichhane, Aashish; Johnson, Carl H; Egli, Martin

    2014-05-01

    A post-translational oscillator (PTO) composed of the proteins KaiA, KaiB and KaiC is at the heart of the cyanobacterial circadian clock. KaiC interacts with KaiA and KaiB over the daily cycle, and CII domains undergo rhythmic phosphorylation/dephosphorylation with a 24 h period. Both the N-terminal (CI) and C-terminal (CII) rings of KaiC exhibit ATPase activity. The CI ATPase proceeds in an input-independent fashion, but the CII ATPase is subject to metabolic input signals. The crystal structure of KaiC from Thermosynechococcus elongatus allows insight into the different anatomies of the CI and CII ATPases. Four consecutive arginines in CI (Arg linker) that connect the P-loop, CI subunits and CI and CII at the ring interface are primary candidates for the coordination of the CI and CII activities. The mutation of linker residues alters the period or triggers arhythmic behavior. Comparison between the CI and CII structures also reveals differences in loop regions that are key to KaiA and KaiB binding and activation of CII ATPase and kinase. Common packing features in KaiC crystals shed light on the KaiB-KaiC interaction. PMID:24816106

  16. Flexibility of the C-terminal, or CII, ring of KaiC governs the rhythm of the circadian clock of cyanobacteria

    PubMed Central

    Chang, Yong-Gang; Kuo, Nai-Wei; Tseng, Roger; LiWang, Andy

    2011-01-01

    In the cyanobacterial circadian oscillator, KaiA and KaiB alternately stimulate autophosphorylation and autodephosphorylation of KaiC with a periodicity of approximately 24 h. KaiA activates autophosphorylation by selectively capturing the A loops of KaiC in their exposed positions. The A loops and sites of phosphorylation, residues S431 and T432, are located in the CII ring of KaiC. We find that the flexibility of the CII ring governs the rhythm of KaiC autophosphorylation and autodephosphorylation and is an example of dynamics-driven protein allostery. KaiA-induced autophosphorylation requires flexibility of the CII ring. In contrast, rigidity is required for KaiC-KaiB binding, which induces a conformational change in KaiB that enables it to sequester KaiA by binding to KaiA’s linker. Autophosphorylation of the S431 residues around the CII ring stabilizes the CII ring, making it rigid. In contrast, autophosphorylation of the T432 residues offsets phospho-S431-induced rigidity to some extent. In the presence of KaiA and KaiB, the dynamic states of the CII ring of KaiC executes the following circadian rhythm: . Apparently, these dynamic states govern the pattern of phosphorylation, ST → SpT → pSpT → pST → ST. CII-CI ring-on-ring stacking is observed when the CII ring is rigid, suggesting a mechanism through which the ATPase activity of the CI ring is rhythmically controlled. SasA, a circadian clock-output protein, binds to the CI ring. Thus, rhythmic ring stacking may also control clock-output pathways. PMID:21788479

  17. Flexibility of the C-terminal, or CII, ring of KaiC governs the rhythm of the circadian clock of cyanobacteria.

    PubMed

    Chang, Yong-Gang; Kuo, Nai-Wei; Tseng, Roger; LiWang, Andy

    2011-08-30

    In the cyanobacterial circadian oscillator, KaiA and KaiB alternately stimulate autophosphorylation and autodephosphorylation of KaiC with a periodicity of approximately 24 h. KaiA activates autophosphorylation by selectively capturing the A loops of KaiC in their exposed positions. The A loops and sites of phosphorylation, residues S431 and T432, are located in the CII ring of KaiC. We find that the flexibility of the CII ring governs the rhythm of KaiC autophosphorylation and autodephosphorylation and is an example of dynamics-driven protein allostery. KaiA-induced autophosphorylation requires flexibility of the CII ring. In contrast, rigidity is required for KaiC-KaiB binding, which induces a conformational change in KaiB that enables it to sequester KaiA by binding to KaiA's linker. Autophosphorylation of the S431 residues around the CII ring stabilizes the CII ring, making it rigid. In contrast, autophosphorylation of the T432 residues offsets phospho-S431-induced rigidity to some extent. In the presence of KaiA and KaiB, the dynamic states of the CII ring of KaiC executes the following circadian rhythm: CII STflexible → CIISpTflexible → CIIpSpTrigid → CIIpSTvery-rigid → CIISTflexible. Apparently, these dynamic states govern the pattern of phosphorylation, ST → SpT → pSpT → pST → ST. CII-CI ring-on-ring stacking is observed when the CII ring is rigid, suggesting a mechanism through which the ATPase activity of the CI ring is rhythmically controlled. SasA, a circadian clock-output protein, binds to the CI ring. Thus, rhythmic ring stacking may also control clock-output pathways. PMID:21788479

  18. Tumor suppressor KAI1 affects integrin {alpha}v{beta}3-mediated ovarian cancer cell adhesion, motility, and proliferation

    SciTech Connect

    Ruseva, Zlatna; Geiger, Pamina Xenia Charlotte; Hutzler, Peter; Kotzsch, Matthias; Luber, Birgit; Schmitt, Manfred; Gross, Eva; Reuning, Ute

    2009-06-10

    The tetraspanin KAI1 had been described as a metastasis suppressor in many different cancer types, a function for which associations of KAI1 with adhesion and signaling receptors of the integrin superfamily likely play a role. In ovarian cancer, integrin {alpha}v{beta}3 correlates with tumor progression and its elevation in vitro provoked enhanced cell adhesion accompanied by significant increases in cell motility and proliferation in the presence of its major ligand vitronectin. In the present study, we characterized integrin {alpha}v{beta}3-mediated tumor biological effects as a function of cellular KAI1 restoration and proved for the first time that KAI1, besides its already known physical crosstalk with {beta}1-integrins, also colocalizes with integrin {alpha}v{beta}3. Functionally, elevated KAI1 levels drastically increased integrin {alpha}v{beta}3/vitronectin-dependent ovarian cancer cell adhesion. Since an intermediate level of cell adhesive strength is required for optimal cell migration, we next studied ovarian cancer cell motility as a function of KAI1 restoration. By time lapse video microscopy, we found impaired integrin {alpha}v{beta}3/vitronectin-mediated cell migration most probably due to strongly enhanced cellular immobilization onto the adhesion-supporting matrix. Moreover, KAI1 reexpression significantly diminished cell proliferation. These data strongly indicate that KAI1 may suppress ovarian cancer progression by inhibiting integrin {alpha}v{beta}3/vitronectin-provoked tumor cell motility and proliferation as important hallmarks of the oncogenic process.

  19. Nm23-H1 was involved in regulation of KAI1 expression in high-metastatic lung cancer cells L9981

    PubMed Central

    Chang, Rui; Liu, Bin; Zu, Lingling

    2016-01-01

    Background The tetraspanin KAI1/CD82 was identified as a tumor metastasis suppressor that down-regulated in malignant progression of lung cancer. However, the underlying mechanism of anti-metastasis role of KAI1 in lung cancer is hardly known. In this paper, we sought to study the function and regulatory mechanism of KAI1 in high metastasis lung cancer cell line. Methods KAI1 expression was detected in high/low metastatic large lung cancer cell line L9981/NL9980 by quantitative real-time polymerase chain reaction (qRT-PCR). The tumor suppressor function of KAI1 was determined by wound healing assay after over-expression or knockdown of KAI1 in L9981 or NL9980 cells. Invasion assay was performed to detect the invasion ability of L9981 by transfection of KAI1. The effect of tumor suppressor p53 on KAI1 expression was measured by western blot and luciferase assay. Then the regulation of KAI1 due to over-expression of metastasis suppressor nm23-H1 was monitored by qRT-PCR, western blot and reporter gene assay. The progression of L9981 cells after p53 and nm23-H1 expression was detected by invasion assay. Also, methylation status of KAI1 promoter in NL9980 and L9981 cells were examined by bisulfite sequencing and methylation-specific PCR. Results We found that KAI1 is down-regulated in high metastatic L9981 cells compare with NL9980 cells. The migration and invasion of L9981 cells were remarkably suppressed in vitro by KAI1 transfection. The migration ability of NL9980 was enhanced by inhibition of KAI1. Furthermore, KAI1 expression was induced after over-expression of p53 or nm23-H1, while cell invasion was inhibited in L9981 cells. The results of reporter analysis indicated that KAI1 promoter region between −922 to −846 could response to nm23-H1. In addition, we discovered only slight methylation of KAI1 promoter, which showed that loss expression of KAI1 in L9981 cells may not due to promoter methylation. Conclusions The results suggested that nm23-H1 was

  20. Diversity of KaiC-based timing systems in marine Cyanobacteria.

    PubMed

    Axmann, Ilka M; Hertel, Stefanie; Wiegard, Anika; Dörrich, Anja K; Wilde, Annegret

    2014-04-01

    The coordination of biological activities into daily cycles provides an important advantage for the fitness of diverse organisms. Most eukaryotes possess an internal clock ticking with a periodicity of about one day to anticipate sunrise and sunset. The 24-hour period of the free-running rhythm is highly robust against many changes in the natural environment. Among prokaryotes, only Cyanobacteria are known to harbor such a circadian clock. Its core oscillator consists of just three proteins, KaiA, KaiB, and KaiC that produce 24-hour oscillations of KaiC phosphorylation, even in vitro. This unique three-protein oscillator is well documented for the freshwater cyanobacterium Synechococcus elongatus PCC 7942. Several physiological studies demonstrate a circadian clock also for other Cyanobacteria including marine species. Genes for the core clock components are present in nearly all marine cyanobacterial species, though there are large differences in the specific composition of these genes. In the first section of this review we summarize data on the model circadian clock from S. elongatus PCC 7942 and compare it to the reduced clock system of the marine cyanobacterium Prochlorococcus marinus MED4. In the second part we discuss the diversity of timing mechanisms in other marine Cyanobacteria with regard to the presence or absence of different components of the clock. PMID:24388874

  1. Structures of KaiC Circadian Clock Mutant Proteins: A New Phosphorylation Site at T426 and Mechanisms of Kinase, ATPase and Phosphatase

    SciTech Connect

    Pattanayek, Rekha; Mori, Tetsuya; Xu, Yao; Pattanayek, Sabuj; Johnson, Carl H.; Egli, Martin

    2010-09-02

    The circadian clock of the cyanobacterium Synechococcus elongatus can be reconstituted in vitro by three proteins, KaiA, KaiB and KaiC. Homo-hexameric KaiC displays kinase, phosphatase and ATPase activities; KaiA enhances KaiC phosphorylation and KaiB antagonizes KaiA. Phosphorylation and dephosphorylation of the two known sites in the C-terminal half of KaiC subunits, T432 and S431, follow a strict order (TS {yields} pTS {yields} pTpS {yields} TpS {yields} TS) over the daily cycle, the origin of which is not understood. To address this void and to analyze the roles of KaiC active site residues, in particular T426, we determined structures of single and double P-site mutants of S. elongatus KaiC. The conformations of the loop region harboring P-site residues T432 and S431 in the crystal structures of six KaiC mutant proteins exhibit subtle differences that result in various distances between Thr (or Ala/Asn/Glu) and Ser (or Ala/Asp) residues and the ATP {gamma}-phosphate. T432 is phosphorylated first because it lies consistently closer to P{gamma}. The structures of the S431A and T432E/S431A mutants reveal phosphorylation at T426. The environments of the latter residue in the structures and functional data for T426 mutants in vitro and in vivo imply a role in dephosphorylation. We provide evidence for a third phosphorylation site in KaiC at T426. T426 and S431 are closely spaced and a KaiC subunit cannot carry phosphates at both sites simultaneously. Fewer subunits are phosphorylated at T426 in the two KaiC mutants compared to phosphorylated T432 and/or S431 residues in the structures of wt and other mutant KaiCs, suggesting that T426 phosphorylation may be labile. The structures combined with functional data for a host of KaiC mutant proteins help rationalize why S431 trails T432 in the loss of its phosphate and shed light on the mechanisms of the KaiC kinase, ATPase and phosphatase activities.

  2. Overexpression of KAI1 induces autophagy and increases MiaPaCa-2 cell survival through the phosphorylation of extracellular signal-regulated kinases

    SciTech Connect

    Wu, Chun-Yan; Yan, Jun; Yang, Yue-Feng; Xiao, Feng-Jun; Li, Qing-Fang; Zhang, Qun-Wei; Wang, Li-Sheng; Guo, Xiao-Zhong; Wang, Hua

    2011-01-21

    Research highlights: {yields} We first investigate the effects of KAI1 on autophagy in MiaPaCa-2 cells. {yields} Our findings demonstrate that KAI1 induces autophagy, which in turn inhibits KAI1-induced apoptosis. {yields} This study also supplies a possible novel therapeutic method for the treatment of pancreatic cancer using autophagy inhibitors. -- Abstract: KAI1, a metastasis-suppressor gene belonging to the tetraspanin family, is known to inhibit cancer metastasis without affecting the primary tumorigenicity by inhibiting the epidermal growth factor (EGF) signaling pathway. Recent studies have shown that hypoxic conditions of solid tumors induce high-level autophagy and KAI1 expression. However, the relationship between autophagy and KAI1 remains unclear. By using transmission electron microscopy, confocal microscopy, and Western blotting, we found that KAI1 can induce autophagy in a dose- and time-dependent manner in the human pancreatic cell line MiaPaCa-2. KAI1-induced autophagy was confirmed by the expression of autophagy-related proteins LC3 and Beclin 1. KAI1 induces autophagy through phosphorylation of extracellular signal-related kinases rather than that of AKT. KAI1-induced autophagy protects MiaPaCa-2 cells from apoptosis and proliferation inhibition partially through the downregulation of poly [adenosine diphosphate (ADP)-ribose] polymerase (PARP) cleavage and caspase-3 activation.

  3. Elevated ATPase activity of KaiC applies a circadian checkpoint on cell division in Synechococcus elongatus.

    PubMed

    Dong, Guogang; Yang, Qiong; Wang, Qiang; Kim, Yong-Ick; Wood, Thammajun L; Osteryoung, Katherine W; van Oudenaarden, Alexander; Golden, Susan S

    2010-02-19

    A circadian clock coordinates physiology and behavior in diverse groups of living organisms. Another major cyclic cellular event, the cell cycle, is regulated by the circadian clock in the few cases where linkage of these cycles has been studied. In the cyanobacterium Synechococcus elongatus, the circadian clock gates cell division by an unknown mechanism. Using timelapse microscopy, we confirm the gating of cell division in the wild-type and demonstrate the regulation of cytokinesis by key clock components. Specifically, a state of the oscillator protein KaiC that is associated with elevated ATPase activity closes the gate by acting through a known clock output pathway to inhibit FtsZ ring formation at the division site. An activity that stimulates KaiC phosphorylation independently of the KaiA protein was also uncovered. We propose a model that separates the functions of KaiC ATPase and phosphorylation in cell division gating and other circadian behaviors. PMID:20178745

  4. Hypersensitive Photic Responses and Intact Genome-Wide Transcriptional Control without the KaiC Phosphorylation Cycle in the Synechococcus Circadian System

    PubMed Central

    Umetani, Miki; Hosokawa, Norimune; Kitayama, Yohko

    2014-01-01

    Cyanobacteria are unique organisms with remarkably stable circadian oscillations. These are controlled by a network architecture that comprises two regulatory factors: posttranslational oscillation (PTO) and a transcription/translation feedback loop (TTFL). The clock proteins KaiA, KaiB, and KaiC are essential for the circadian rhythm of the unicellular species Synechococcus elongatus PCC 7942. Temperature-compensated autonomous cycling of KaiC phosphorylation has been proposed as the primary oscillator mechanism that maintains the circadian clock, even in the dark, and it controls genome-wide gene expression rhythms under continuous-light conditions (LL). However, the kaiCEE mutation (where “EE” represents the amino acid changes Ser431Glu and Thr432Glu), where phosphorylation cycling does not occur in vivo, has a damped but clear kaiBC expression rhythm with a long period. This suggests that there must be coupling between the robust PTO and the “slave” unstable TTFL. Here, we found that the kaiCEE mutant strain in LL was hypersensitive to the dark acclimation required for phase shifting. Twenty-three percent of the genes in the kaiCEE mutant strain exhibited genome-wide transcriptional rhythms with a period of 48 h in LL. The circadian phase distribution was also conserved significantly in most of the wild-type and kaiCEE mutant strain cycling genes, which suggests that the output mechanism was not damaged severely even in the absence of KaiC phosphorylation cycles. These results strongly suggest that the KaiC phosphorylation cycle is not essential for generating the genome-wide rhythm under light conditions, whereas it is important for appropriate circadian timing in the light and dark. PMID:24244001

  5. 75 FR 35513 - Application of Schuman Aviation Company Ltd. D/B/A Makani Kai Helicopters D/B/A Ko Olina...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-06-22

    ... Office of the Secretary Application of Schuman Aviation Company Ltd. D/B/A Makani Kai Helicopters D/B/A Ko Olina Helicopters D/B/A Pacific Air Express D/B/A Makani Kai Air Charters; For Commuter Authority... cause why it should not issue an order finding Schuman Aviation Company Ltd. d/b/a Makani...

  6. The Use of Rockets as Military Weapons at the Siege of Kai Fung Foo in 1232 A.D

    NASA Technical Reports Server (NTRS)

    2004-01-01

    One of the earliest recorded instances of the use of rockets was as military weapons against the Mongols by the Chinese at the siege of Kai Fung Foo in 1232 A.D. An arrow with a tube of gunpowder produced an arrow of flying fire. The Mongol attackers fled in terror, even though the rockets were inaccurate and relatively harmless.

  7. The Academic Profession and University Governance Participation in Japan: Focusing on the Role of Kyoju-kai

    ERIC Educational Resources Information Center

    Yonezawa, Akiyoshi

    2014-01-01

    The dominant role of Kyoju-kai (the professoriate) in university governance in Japan is now facing a critical examination as part of university reforms in response to global competition. What are the determinants of the characteristics of participation in university governance by individual faculty members? In what way does the organizational…

  8. Hui Malama O Ke Kai: A Positive Prevention-Based Youth Development Program Based on Native Hawaiian Values and Activities

    ERIC Educational Resources Information Center

    Hishinuma, Earl S.; Chang, Janice Y.; Sy, Angela; Greaney, Malia F.; Morris, Katherine A.; Scronce, Ami C.; Rehuher, Davis; Nishimura, Stephanie T.

    2009-01-01

    Evaluation of after-school programs that are culturally and place-based and promote positive youth development among minority and indigenous youths has not been widely published. The present evaluation is the first of its kind of an after-school, youth-risk prevention program called Hui Malama O Ke Kai (HMK), that emphasizes Native Hawaiian values…

  9. Dissecting the Diverse Functions of the Metastasis Suppressor CD82/KAI1

    PubMed Central

    Tsai, Yien Che; Weissman, Allan M.

    2011-01-01

    The recent identification of metastasis suppressor genes, the products of which inhibit metastasis but not primary tumor growth, distinguishes oncogenic transformation and tumor suppression from a hallmark of malignancy, the ability of cancer cells to invade sites distant from the primary tumor. The metastasis suppressor CD82/KAI1 is a member of the tetraspanin superfamily of glycoproteins. CD82 suppresses metastasis by multiple mechanisms including inhibition of cell motility and invasion, promotion of cell polarity as well as induction of senescence and apoptosis in response to extracellular stimuli. A common feature of these diverse effects is CD82 regulation of membrane organization as well as protein trafficking and interactions, which affects cellular signaling and intercellular communication. PMID:21875585

  10. Evolution of KaiC-Dependent Timekeepers: A Proto-circadian Timing Mechanism Confers Adaptive Fitness in the Purple Bacterium Rhodopseudomonas palustris

    PubMed Central

    Ma, Peijun; Mori, Tetsuya; Zhao, Chi; Thiel, Teresa; Johnson, Carl Hirschie

    2016-01-01

    Circadian (daily) rhythms are a fundamental and ubiquitous property of eukaryotic organisms. However, cyanobacteria are the only prokaryotic group for which bona fide circadian properties have been persuasively documented, even though homologs of the cyanobacterial kaiABC central clock genes are distributed widely among Eubacteria and Archaea. We report the purple non-sulfur bacterium Rhodopseudomonas palustris (that harbors homologs of kaiB and kaiC) only poorly sustains rhythmicity in constant conditions–a defining characteristic of circadian rhythms. Moreover, the biochemical characteristics of the Rhodopseudomonas homolog of the KaiC protein in vivo and in vitro are different from those of cyanobacterial KaiC. Nevertheless, R. palustris cells exhibit adaptive kaiC-dependent growth enhancement in 24-h cyclic environments, but not under non-natural constant conditions. Therefore, our data indicate that Rhodopseudomonas does not have a classical circadian rhythm, but a novel timekeeping mechanism that does not sustain itself in constant conditions. These results question the adaptive value of self-sustained oscillatory capability for daily timekeepers and establish new criteria for circadian-like systems that are based on adaptive properties (i.e., fitness enhancement in rhythmic environments), rather than upon observations of persisting rhythms in constant conditions. We propose that the Rhodopseudomonas system is a "proto" circadian timekeeper, as in an ancestral system that is based on KaiC and KaiB proteins and includes some, but not necessarily all, of the canonical properties of circadian clocks. These data indicate reasonable intermediate steps by which bona fide circadian systems evolved in simple organisms. PMID:26982486

  11. KAI407, a Potent Non-8-Aminoquinoline Compound That Kills Plasmodium cynomolgi Early Dormant Liver Stage Parasites In Vitro

    PubMed Central

    Zeeman, Anne-Marie; van Amsterdam, Sandra M.; McNamara, Case W.; Voorberg-van der Wel, Annemarie; Klooster, Els J.; van den Berg, Alexander; Remarque, Edmond J.; Plouffe, David M.; van Gemert, Geert-Jan; Luty, Adrian; Sauerwein, Robert; Gagaring, Kerstin; Borboa, Rachel; Chen, Zhong; Kuhen, Kelli; Glynne, Richard J.; Chatterjee, Arnab K.; Nagle, Advait; Roland, Jason; Winzeler, Elizabeth A.; Leroy, Didier; Campo, Brice; Diagana, Thierry T.; Yeung, Bryan K. S.; Thomas, Alan W.

    2014-01-01

    Preventing relapses of Plasmodium vivax malaria through a radical cure depends on use of the 8-aminoquinoline primaquine, which is associated with safety and compliance issues. For future malaria eradication strategies, new, safer radical curative compounds that efficiently kill dormant liver stages (hypnozoites) will be essential. A new compound with potential radical cure activity was identified using a low-throughput assay of in vitro-cultured hypnozoite forms of Plasmodium cynomolgi (an excellent and accessible model for Plasmodium vivax). In this assay, primary rhesus hepatocytes are infected with P. cynomolgi sporozoites, and exoerythrocytic development is monitored in the presence of compounds. Liver stage cultures are fixed after 6 days and stained with anti-Hsp70 antibodies, and the relative proportions of small (hypnozoite) and large (schizont) forms relative to the untreated controls are determined. This assay was used to screen a series of 18 known antimalarials and 14 new non-8-aminoquinolines (preselected for blood and/or liver stage activity) in three-point 10-fold dilutions (0.1, 1, and 10 μM final concentrations). A novel compound, designated KAI407 showed an activity profile similar to that of primaquine (PQ), efficiently killing the earliest stages of the parasites that become either primary hepatic schizonts or hypnozoites (50% inhibitory concentration [IC50] for hypnozoites, KAI407, 0.69 μM, and PQ, 0.84 μM; for developing liver stages, KAI407, 0.64 μM, and PQ, 0.37 μM). When given as causal prophylaxis, a single oral dose of 100 mg/kg of body weight prevented blood stage parasitemia in mice. From these results, we conclude that KAI407 may represent a new compound class for P. vivax malaria prophylaxis and potentially a radical cure. PMID:24366744

  12. Spent Mushroom Waste as a Media Replacement for Peat Moss in Kai-Lan (Brassica oleracea var. Alboglabra) Production

    PubMed Central

    Sendi, H.; Mohamed, M. T. M.; Anwar, M. P.; Saud, H. M.

    2013-01-01

    Peat moss (PM) is the most widely used growing substrate for the pot culture. Due to diminishing availability and increasing price of PM, researchers are looking for viable alternatives for peat as a growth media component for potted plants. A pot study was conducted with a view to investigate the possibility of using spent mushroom waste (SMW) for Kai-lan (Brassica oleracea var. Alboglabra) production replacing peat moss (PM) in growth media. The treatments evaluated were 100% PM (control), 100% SMW, and mixtures of SMW and PM in different ratios like 1 : 1, 1 : 2, and 2 : 1 (v/v) with/without NPK amendment. The experiment was arranged in a completely randomized design with five replications per treatment. Chemical properties like pH and salinity level (EC) of SMW were within the acceptable range of crop production but, nutrient content, especially nitrogen content was not enough to provide sufficient nutrition to plant for normal growth. Only PM (100%) and SMW and PM mixture in 1 : 1 ratio with NPK amendment performed equally in terms of Kai-lan growth. This study confirms the feasibility of replacing PM by SMW up to a maximum of 50% in the growth media and suggests that NPK supplementation from inorganic sources is to ensure a higher productivity of Kai-lan. PMID:24106452

  13. Genetic diversity and population structure of ‘Khao Kai Noi’, a Lao rice (Oryza sativa L.) landrace, revealed by microsatellite DNA markers

    PubMed Central

    Vilayheuang, Koukham; Machida-Hirano, Ryoko; Bounphanousay, Chay; Watanabe, Kazuo N.

    2016-01-01

    Rice (Oryza sativa L.) is the main food for people in Laos, where it has been grown and eaten since prehistory. Diverse landraces are grown in Laos. ‘Khao Kai Noi’, a landrace favored for its eating quality, is held in the nationwide collection of traditional landraces in the Lao national genebank. Genetic diversity is crucial for sustainable use of genetic resources and conservation. To investigate the genetic diversity of ‘Khao Kai Noi’ for conservation, we genotyped 70 accessions by using 23 polymorphic simple sequence repeat markers. The markers generated 2 to 17 alleles (132 in total), with an average of 5.7 per locus. The total expected heterozygosity over all ‘Khao Kai Noi’ accessions was 0.271. Genetic variation was largest among accessions and smallest within accessions. Khao Kai Noi accessions were classified into three different genetic backgrounds, but there was unclear association between the three inferred population and name subgroups and geographical distribution. Most of the accessions were clustered with temperate japonica and showed genetic relatedness to rice from neighboring provinces of Vietnam, suggesting a Vietnamese origin. The results of this study will contribute to the conservation, core collection and future breeding of the Khao Kai Noi population. PMID:27162492

  14. Deletion of the Synechocystis sp. PCC 6803 kaiAB1C1 gene cluster causes impaired cell growth under light-dark conditions.

    PubMed

    Dörrich, Anja K; Mitschke, Jan; Siadat, Olga; Wilde, Annegret

    2014-11-01

    In contrast to Synechococcus elongatus PCC 7942, few data exist on the timing mechanism of the widely used cyanobacterium Synechocystis sp. PCC 6803. The standard kaiAB1C1 operon present in this organism was shown to encode a functional KaiC protein that interacted with KaiA, similar to the S. elongatus PCC 7942 clock. Inactivation of this operon in Synechocystis sp. PCC 6803 resulted in a mutant with a strong growth defect when grown under light-dark cycles, which was even more pronounced when glucose was added to the growth medium. In addition, mutants showed a bleaching phenotype. No effects were detected in mutant cells grown under constant light. Microarray experiments performed with cells grown for 1 day under a light-dark cycle revealed many differentially regulated genes with known functions in the ΔkaiABC mutant in comparison with the WT. We identified the genes encoding the cyanobacterial phytochrome Cph1 and the light-repressed protein LrtA as well as several hypothetical ORFs with a complete inverse behaviour in the light cycle. These transcripts showed a stronger accumulation in the light but a weaker accumulation in the dark in ΔkaiABC cells in comparison with the WT. In general, we found a considerable overlap with microarray data obtained for hik31 and sigE mutants. These genes are known to be important regulators of cell metabolism in the dark. Strikingly, deletion of the ΔkaiABC operon led to a much stronger phenotype under light-dark cycles in Synechocystis sp. PCC 6803 than in Synechococcus sp. PCC 7942. PMID:25139948

  15. Early diagenetic processes of saline meromictic Lake Kai-ike, southwest Japan: III. Sulfur speciation and isotopes

    NASA Astrophysics Data System (ADS)

    Sakai, N.; Yamaguchi, K. E.; Oguri, K.

    2014-12-01

    Lake Kai-ike is a saline meromictic lake located along the coast of Kami-Koshiki Island. The lake is isolated from ocean by a gravel bar, through which seawater infiltrates by tidal pumping. The lake is permanently redox (density)-stratified with a mid-depth development of photic zone anoxia and a dense community of photosynthetic bacteria pinkish "bacterial plate". The early diagenesis of sulfur in sediments overlain by an anoxic water body was investigated using a sediment core (KAI4) from the lake. We determined abundance of various S-bearing species (i.e., Cr-reducible sulfide (= pyrite S: Spy), acid-volatile sulfide (AVS), sulfate sulfur (SSO4), elemental sulfur (S0), and organic sulfur) by an improved sequential extraction method. Here we focus on drastic and rapid changes on sulfur biogeochemistry found in the uppermost 5cm layer. With increasing depth, abundance of Spy increased but that of SSO4 and δ34S value of Spy (δ34Spy) decreased. These results suggest progressive formation of bacteriogenic pyrite. The δ34S values of SSO4 (δ34SSO4) ranged from 25.1 ‰ (at sediment surface) to 3.8 ‰ in the uppermost 5 cm layer. This δ34SSO4 decrease in the top 5 cm sediment suggests that SSO4 in the surface sediment inherits SO42- with elevated δ34S values (higher than typical seawater δ34S value of 21‰) in the water column, which is due to extensive bacterial sulfate reduction with preferential removal of low-δ34S sulfur as sulfide. In the lower part of the uppermost 5 cm layer, SO42- formed by oxidation of S0, AVS, and/or Spy with low-δ34S values by SO42--bearing seawater introduced by infiltration through the gravel bar. Increasing δ34Spy values with increasing depth suggest near complete consumption of SO42- by active bacterial sulfate reduction, and this process could be explained by Rayleigh distillation model. Early diagenesis of sulfur does occur in whole section of 25cm-long KAI4 core that accumulated for the last ~60 years (Yamaguchi et al

  16. Phosphorus and iron cycles during early diagenesis of Lake Kai-ike sediments, Kami-koshiki Island, southwest Japan

    NASA Astrophysics Data System (ADS)

    Iida, H.; Yamaguchi, K. E.; Oguri, K.

    2014-12-01

    A meromictic Lake Kai-ike is located on the northeastern margin of Kami-koshiki island, Japan. Permanent density stratification develops due to seawater infiltration through a gravel bar separating the lake from the ocean. The oxygenated surface water overlays a stagnant, saline, and anoxic deep water containing hydrogen sulfide. Purple sulfur bacteria (Chromatium sp.) inhabit the chemocline at 4.5m depth. At the lake bottom, green sulfur bacteria form microbial mat-like structures (Nakajima et al., 2003; Environ. Microbiol.). Such environment can be treated as a model for the past anoxic ocean, such as during Cretaceous OAEs (Oguri et al., 2003; Frontier Res. on Earth Evol.). A 25 cm-long KAI4 sediment core (Yamaguchi et al., 2010; Palaeo3) was used for two sequential extraction methods. SEDEX method (Ruttenberg, 1992; Limnol. Oceanogr.) was used for partitioning phosphorus-bearing species into Pabs (absorbed), PFe (Fe-bound), Pauth (authigenic), Pdet (detrital), and Porg (organic). Iron-bearing species were also divided into FeHCl (HCl-soluble), Fecarb (carbonate), Feox (oxide), Femag (magnetite), and Feresi (residue), following the method of Poulton et al. (2005; Chem. Geol.). At the uppermost part of KAI4 core, Porg was the most abundant P-bearing species (~90% of total P). The Porg content sharply decreased with increasing depth to 5cm. The second most abundant species was PFe; however, PFe and Feox contents remained constant throughout the whole depth. At sediment surface in present-day oxygenated ocean, Fe3+-(oxy)hydroxides trap phosphate diffusing from deeper-anoxic sediment, and the phosphate concentration in pore water becomes high enough to precipitate authigenic apatite (Slomp et al., 1996; J. Mar. Res.). In case of Lake Kai-ike, however, the amount of Fe3+-(oxy)hydroxides was small relative to that of Porg (PFe/Porg = ~0.1). We suggest that the excess phosphate not adsorbed on Fe3+-(oxy)hydroxides was diffused out to the overlying water mass, and the

  17. Characterization of multiple constituents in Kai-Xin-San prescription and rat plasma after oral administration by liquid chromatography with quadrupole time-of-flight tandem mass spectrometry.

    PubMed

    Zhang, Xiaowen; Li, Qing; Lv, Chunxiao; Xu, Huarong; Liu, Xujia; Sui, Zhenyu; Bi, Kaishun

    2015-06-01

    A sensitive and reliable ultra high performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry method was established to separate and identify the chemical constituents of Kai-Xin-San prescription, a classic traditional Chinese medicine formula that plays an important role in the treatment of Alzheimer's disease. The detection was performed on an Agilent 6520 Accurate-Mass quadrupole time-of-flight mass spectrometer equipped with an electrospray ionization source in negative modes. With the optimized conditions, a total of 54 compounds were identified or tentatively characterized. Out of the 54 compounds, six compounds were identified by comparing the retention time and mass spectrometry data with reference standards, the rest were characterized by analyzing mass spectrometry data and retrieving the literature data. Results indicated ginsenosides, polygala saponins, terpenoids, and oligosaccharide esters were the major effective constituents in Kai-Xin-San prescription. There were 26 prototype ingredients that were assigned for identification in rat plasma. It is also concluded that the developed ultra high performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry method with high sensitivity and resolution is suitable for identifying and characterizing the chemical constituents of Kai-Xin-San prescription, and the analysis provides a helpful chemical basis for further research on Kai-Xin-San prescription and the clinical diagnostics of Alzheimer's disease. PMID:25845859

  18. The Experience of KAI MAHI, an Employment Initiative for People with an Experience of Mental Illness, as Told by Zarna, Zeus, Lulu, Mary, Paul, and Hemi.

    PubMed

    de Malmanche, Janie; Robertson, Linda

    2015-11-01

    There is a growing body of evidence endorsing the use of supported employment models for people with an experience of mental illness. However current literature and research regarding the perspective of people with experience of mental illness, as well as alternate models of employment support, is sparse. This study has captured the stories of employment of people with experience of mental illness who participated in KAI MAHI, a group based employment program. Findings identified key components of KAI MAHI, consistent with their overall experiences of employment, which were influential in assisting them to find and sustain employment. These included the opportunity for self-determination, a sense of self-efficacy, and respectful relationships. PMID:26173402

  19. Behavioral and biochemical effects of a formulation of the traditional Chinese medicine, Kai-Xin-San, in fatigued rats

    PubMed Central

    HU, YUAN; CAO, YIN; LIU, MING; LIU, PING; CUI, HONG; DAI-HONG, GUO

    2013-01-01

    The present study was designed to evaluate the anti-fatigue activity and the behavioral and biochemical effects of Kai-Xin-San (KXS) extracts on fatigued rats. The rats were randomly divided into six groups: untreated control (UC), running control (RC), RC treated with 13 mg/kg/day modafinil and RC treated with KXS at dosages of 125, 250 and 500 mg/kg/day, respectively. The treatments were administered orally. Anti-fatigue activity was assessed using the treadmill running test and serum biochemical parameters were determined using an autoanalyzer and commercially available kits. Furthermore, the standardization of the KXS extracts was ensured using a high-performance liquid chromatography (HPLC)-fingerprint. The extracts were shown to increase exhaustive running time in the treadmill running test and reverse the fatigue-induced reduction in hepatic/muscle glycogen and testosterone, in addition to reducing the lactate dehydrogenase (LDH), serum urea nitrogen (SUN), blood lactic acid (BLA) and β-endorphin levels in the serum of the fatigued rats. Moreover, the extracts enhanced superoxide dismutase (SOD) activity and decreased the malondialdehyde (MDA) levels in the serum of the fatigued rats. The results of this preliminary study indicated that KXS exhibits anti-fatigue activity. This was reflected in the effects on the biochemical markers for fatigue. PMID:24137300

  20. Expression, purification, crystallization and preliminary X-ray analysis of the KaiC-like protein PH0187 from the hyperthermophilic archaeon Pyrococcus horikoshii OT3

    PubMed Central

    Kang, Hee-Jin; Kubota, Keiko; Miyazono, Ken-ichi; Tanokura, Masaru

    2011-01-01

    KaiC is the central protein in the circadian rhythm in cyanobacteria. The 28 kDa KaiC-like protein PH0187 from the hyperthermophilic archaeon Pyrococcus horikoshii was expressed in Escherichia coli, purified and crystallized using the sitting-drop vapour-diffusion method at 293 K. Crystals of PH0187 were obtained using a reservoir solution consisting of 1.0 M ammonium phosphate monobasic and 0.1 M sodium citrate tribasic pH 5.3 (the final pH value of the reservoir solution was 4.8) and diffracted X-rays to 2.75 Å resolution. The crystal of PH0187 belonged to space group P6322, with unit-cell parameters a = b = 239.1, c = 106.5 Å. The crystal contained four PH0187 molecules in the asymmetric unit. PMID:21206047

  1. A comitative source for object markers in Sinitic languages: 跟 kai55 in Waxiang and 共 kang7 in Southern Min

    PubMed Central

    Chappell, Hilary; Peyraube, Alain; Wu, Yunji

    2013-01-01

    This analysis sets out to specifically discuss the polyfunctionality of 跟 [kai55] in Waxiang (Sinitic), whose lexical source is the verb ‘to follow’. Amongst its various uses, we find a preposition ‘with, along’, a marker of adjuncts and a NP conjunction, thus superficially resembling its Mandarin cognate 跟 gēn ‘with’. Curiously, however, it has also evolved into a direct object marker in Waxiang, with a function similar to that of preposition 把 bă < ‘hold, take’ as found in the S–bă–O–VP or so-called ‘disposal’ form in standard Mandarin. The pathways of grammaticalization for 跟 [kai55] in Waxiang are thus discussed in order to determine how it has developed this unusual grammatical function in one of the linguistic zones of China where verbs of giving or taking are, in fact, the main source for grammaticalized object markers in ‘disposal’ constructions. On the basis of 16th and 17th century Southern Min literature (Sinitic), a comparison is also made with analogous developments for comitative 共 gòng ‘with’ to provide support for our hypothesis that the direct object marking use has evolved from the oblique function of a benefactive or dative, and is clearly separate from the crosslinguistically well-attested pathway that leads to its use as a conjunction. We would thus like to propose that these data contribute a new pattern to the stock of grammaticalization pathways, specifically, comitative > dative/benefactive > accusative (direct object marker). PMID:24273384

  2. A Chinese herbal decoction, reformulated from Kai-Xin-San, relieves the depression-like symptoms in stressed rats and induces neurogenesis in cultured neurons

    PubMed Central

    Yan, Lu; Hu, Qinghua; Mak, Marvin S. H.; Lou, Jianshu; Xu, Sherry L.; Bi, Cathy W. C.; Zhu, Yue; Wang, Huaiyou; Dong, Tina T. X.; Tsim, Karl W. K.

    2016-01-01

    Kai-Xin-San (KXS), a Chinese herbal decoction for anti-depression, is a combination of paired-herbs, i.e. Ginseng Radix et Rhizoma (GR)-Polygalae Radix (PR) and Acori Tatarinowii Rhizoma (ATR)-Poria (PO). The make-up of the paired-herbs has been commonly revised according to syndrome differentiation and treatment variation of individual. Currently, an optimized KXS (KXS2012) was prepared by functional screening different combination of GR-PR and ATR-PO. The aim of this study was to verify the effect and underlying mechanism of KXS2012 against depression in chronic mild stress (CMS)-induced depressive rats and in primary cultures of neurons and astrocytes. In rat model, the CMS-induced depressive symptoms were markedly alleviated by the treatment with KXS2012. The CMS-suppressed neurotransmitter amounts were restored in the presence of KXS2012. And the expressions of neurotropic factors and its corresponding receptors were increased under KXS2012 administration. In cultured neurons, application of KXS2012 could promote neurogenesis by inducing the expression of synaptotagmin and dendritic spine density. Moreover, application of KXS2012 in cultured astrocytes, or in H2O2-stressed astrocytes, induced the expressions of neurotrophic factors: the increase might be associated with the modification of Erk1/2 and CREB phosphorylation. Our current results fully support the therapeutic efficacy of KXS2012 against depression in cell and animal models. PMID:27444820

  3. A Chinese herbal decoction, reformulated from Kai-Xin-San, relieves the depression-like symptoms in stressed rats and induces neurogenesis in cultured neurons.

    PubMed

    Yan, Lu; Hu, Qinghua; Mak, Marvin S H; Lou, Jianshu; Xu, Sherry L; Bi, Cathy W C; Zhu, Yue; Wang, Huaiyou; Dong, Tina T X; Tsim, Karl W K

    2016-01-01

    Kai-Xin-San (KXS), a Chinese herbal decoction for anti-depression, is a combination of paired-herbs, i.e. Ginseng Radix et Rhizoma (GR)-Polygalae Radix (PR) and Acori Tatarinowii Rhizoma (ATR)-Poria (PO). The make-up of the paired-herbs has been commonly revised according to syndrome differentiation and treatment variation of individual. Currently, an optimized KXS (KXS2012) was prepared by functional screening different combination of GR-PR and ATR-PO. The aim of this study was to verify the effect and underlying mechanism of KXS2012 against depression in chronic mild stress (CMS)-induced depressive rats and in primary cultures of neurons and astrocytes. In rat model, the CMS-induced depressive symptoms were markedly alleviated by the treatment with KXS2012. The CMS-suppressed neurotransmitter amounts were restored in the presence of KXS2012. And the expressions of neurotropic factors and its corresponding receptors were increased under KXS2012 administration. In cultured neurons, application of KXS2012 could promote neurogenesis by inducing the expression of synaptotagmin and dendritic spine density. Moreover, application of KXS2012 in cultured astrocytes, or in H2O2-stressed astrocytes, induced the expressions of neurotrophic factors: the increase might be associated with the modification of Erk1/2 and CREB phosphorylation. Our current results fully support the therapeutic efficacy of KXS2012 against depression in cell and animal models. PMID:27444820

  4. A Standardized Chinese Herbal Decoction, Kai-Xin-San, Restores Decreased Levels of Neurotransmitters and Neurotrophic Factors in the Brain of Chronic Stress-Induced Depressive Rats

    PubMed Central

    Zhu, Kevin Yue; Mao, Qing-Qiu; Ip, Siu-Po; Choi, Roy Chi-Yan; Dong, Tina Ting-Xia; Lau, David Tai-Wai; Tsim, Karl Wah-Keung

    2012-01-01

    Kai-xin-san (KXS), a Chinese herbal decoction being prescribed by Sun Simiao in Beiji Qianjin Yaofang about 1400 years ago, contains Ginseng Radix et Rhizoma, Polygalae Radix, Acori tatarinowii Rhizoma, and Poria. KXS has been used to treat stress-related psychiatric disease with the symptoms of depression and forgetfulness in ancient China until today. However, the mechanism of its antidepression action is still unknown. Here, the chronic mild-stress-(CMS-) induced depressive rats were applied in exploring the action mechanisms of KXS treatment. Daily intragastric administration of KXS for four weeks significantly alleviated the CMS-induced depressive symptoms displayed by enhanced sucrose consumption. In addition, the expressions of those molecular bio-markers relating to depression in rat brains were altered by the treatment of KXS. These KXS-regulated brain biomarkers included: (i) the levels of dopamine, norepinephrine, and serotonin (ii) the transcript levels of proteins relating to neurotransmitter metabolism; (iii) the transcript levels of neurotrophic factors and their receptors. The results suggested that the anti-depressant-like action of KXS might be mediated by an increase of neurotransmitters and expression of neurotrophic factors and its corresponding receptors in the brain. Thus, KXS could serve as alternative medicine, or health food supplement, for patients suffering from depression. PMID:22973399

  5. Synergism and Rules from Combination of Baicalin, Jasminoidin and Desoxycholic acid in Refined Qing Kai Ling for Treat Ischemic Stroke Mice Model

    PubMed Central

    Wang, Zhong; Wang, Chang-ming; Wang, Yong-yan; Zhang, Zhan-jun

    2012-01-01

    Refined Qing-Kai-Ling (QKL), a modified Chinese medicine, consists of three main ingredients (Baicalin, Jasminoidin and Desoxycholic acid), plays a synergistic effect on the treatment of the acute stage of ischemic stroke. However, the rules of the combination and synergism are still unknown. Based on the ischemic stroke mice model, all different kinds of combination of Baicalin, Jasminoidin, and Desoxycholic acid were investigated by the methods of neurological examination, microarray, and genomics analysis. As a result, it confirmed that the combination of three drugs offered a better therapeutical effect on ischemic stroke than monotherapy of each drug. Additionally, we used Ingenuity pathway Analysis (IPA) and principal component analysis (PCA) to extract the dominant information of expression changes in 373 ischemia-related genes. The results suggested that 5 principal components (PC1-5) could account for more than 95% energy in the gene data. Moreover, 3 clusters (PC1, PC2+PC5, and PC3+PC4) were addressed with cluster analysis. Furthermore, we matched PCs on the drug-target networks, the findings demonstrated that Baicalin related with PC1 that played the leading role in the combination; Jasminoidin related with PC2+PC5 that played a compensatory role; while Desoxycholic acid had the least performance alone which could relate with PC3+PC4 that played a compatible role. These manifestations were accorded with the principle of herbal formulae of Traditional Chinese Medicine (TCM), emperor-minister-adjuvant-courier. In conclusion, we firstly provided scientific evidence to the classic theory of TCM formulae, an initiating holistic viewpoint of combination therapy of TCM. This study also illustrated that PCA might be an applicable method to analyze the complicated data of drug combination. PMID:23049867

  6. The tetraspanin, KAI1/CD82, is expressed by late-lineage oligodendrocyte precursors and may function to restrict precursor migration and promote oligodendrocyte differentiation and myelination

    PubMed Central

    Mela, Angeliki; Goldman, James E.

    2009-01-01

    In the adult mammalian brain, oligodendrocyte progenitors can differentiate into mature oligodendrocytes during remyelination. Mechanisms that regulate migration and differentiation of progenitors are of great importance in understanding normal development and demyelinating/remyelinating conditions. In a microarray analysis comparing adult and neonatal O4+ cells, we found that the tetraspanin, KAI1/CD82, is far more highly expressed in adult O4+ cells than in neonatal O4+ cells (Lin et al., in press). CD82 is a metastasis suppressor and its expression is often down-regulated or lost in the advanced stages of metastatic cancer. We hypothesized that CD82 could be a factor that restricts migration and promotes differentiation of maturing oligodendrocytes. Western analysis of isolated adult O4+ cells confirms the elevated levels of CD82, which continues to be expressed as these become O1+ in vitro. In the adult rat white matter CD82 is co-expressed with CC1 and olig2 but not with NG2 or GFAP. Immature cells of the neonatal forebrain subventricular zone (SVZ) infected in vivo with a retrovirus that constitutively expresses CD82 do not remain immature, but differentiate either into CC1+ and MBP+ myelinating oligodendrocytes in the white matter or zebrinII+ astrocytes in the cortex. Their migration from the SVZ is severely restricted. In contrast, downregulation of CD82 in SVZ cells in vivo, using retroviral-expressed shRNAs, prevents their differentiation into myelinating oligodendrocytes. shRNA-expressing cells remained PDGFRα+, olig2+ or NG2+, or became CC1+ non-myelinating oligodendrocytes, or GFAP+ astrocytes. CD82 thus appears to be a critical molecule in the regulation of oligodendrocyte progenitor migration and myelination. PMID:19741124

  7. Synergism and rules from combination of Baicalin, Jasminoidin and Desoxycholic acid in refined Qing Kai Ling for treat ischemic stroke mice model.

    PubMed

    Li, Jian; Wu, Run-guo; Meng, Fan-yun; Wang, Zhong; Wang, Chang-ming; Wang, Yong-yan; Zhang, Zhan-jun

    2012-01-01

    Refined Qing-Kai-Ling (QKL), a modified Chinese medicine, consists of three main ingredients (Baicalin, Jasminoidin and Desoxycholic acid), plays a synergistic effect on the treatment of the acute stage of ischemic stroke. However, the rules of the combination and synergism are still unknown. Based on the ischemic stroke mice model, all different kinds of combination of Baicalin, Jasminoidin, and Desoxycholic acid were investigated by the methods of neurological examination, microarray, and genomics analysis. As a result, it confirmed that the combination of three drugs offered a better therapeutical effect on ischemic stroke than monotherapy of each drug. Additionally, we used Ingenuity pathway Analysis (IPA) and principal component analysis (PCA) to extract the dominant information of expression changes in 373 ischemia-related genes. The results suggested that 5 principal components (PC1-5) could account for more than 95% energy in the gene data. Moreover, 3 clusters (PC1, PC2+PC5, and PC3+PC4) were addressed with cluster analysis. Furthermore, we matched PCs on the drug-target networks, the findings demonstrated that Baicalin related with PC1 that played the leading role in the combination; Jasminoidin related with PC2+PC5 that played a compensatory role; while Desoxycholic acid had the least performance alone which could relate with PC3+PC4 that played a compatible role. These manifestations were accorded with the principle of herbal formulae of Traditional Chinese Medicine (TCM), emperor-minister-adjuvant-courier. In conclusion, we firstly provided scientific evidence to the classic theory of TCM formulae, an initiating holistic viewpoint of combination therapy of TCM. This study also illustrated that PCA might be an applicable method to analyze the complicated data of drug combination. PMID:23049867

  8. Identification and determination of the major constituents in Kai-Xin-San by UPLC-Q/TOF MS and UFLC-MS/MS method.

    PubMed

    Lv, Chunxiao; He, Bosai; Sui, Zhenyu; Li, Qing; Bi, Kaishun

    2016-07-01

    In order to have overall chemical material information of Kai-Xin-San (KXS), the reliable ultra-high-performance liquid chromatography quadrupole time-of-flight mass spectrometer (UHPLC-Q-TOF-MS) and ultra-fast liquid chromatography mass spectrometer (UFLC-MS/MS) methods were developed for the identification and determination of the major constituents in KXS. Moreover, the UHPLC-Q-TOF-MS method was also applied to screen for multiple absorbed components in rat plasma after oral administration of KXS. The UHPLC-Q-TOF-MS method was achieved on Agilent 6520 Q-TOF mass and operated in the negative ion mode. Good separation was performed on a ZORBAX Eclipse Plus C18 column with a gradient elution at a flow rate of 0.2 ml/min. A total of 92 compounds in KXS were identified or tentatively characterized based on their exact molecular weights, fragmentation patterns, and literature data. A total of 26 compounds including 23 prototype components and three metabolites were identified in rat plasma after oral administration of KXS. Then, 16 major bioactive constituents were chosen as the benchmark substances to evaluate the quality of KXS. Their quantitative analyses were performed by a triple quadrupole tandem mass spectrometer (MS/MS) operating in multiple-reaction monitoring mode(MRM). The analysis was completed with a gradient elution at a flow rate of 0.4 ml/min within 35 min. The simple and fast method was validated and showed good linearity, precision, and recovery. Furthermore, the method was successful applied for the determination of 16 compounds in KXS. All results would provide essential data for identification and quality control of active chemical constituents in KXS. Copyright © 2016 John Wiley & Sons, Ltd. PMID:27434806

  9. Metabolomics approach to explore the effects of Kai-Xin-San on Alzheimer's disease using UPLC/ESI-Q-TOF mass spectrometry.

    PubMed

    Chu, Hang; Zhang, Aihua; Han, Ying; Lu, Shengwen; Kong, Ling; Han, Jinwei; Liu, Zhidong; Sun, Hui; Wang, Xijun

    2016-03-15

    Alzheimer's disease (AD) is a multifactorial neurodegenerative disease that influences elderly populations, with no effective method for its treatment so far. To improve its diagnosis and treatment, changes of small molecule metabolite during AD should be elucidated. Kai-Xin-San (KXS) is an herbal formulae that has been widely used to treat mental disorders, especially amnesia and depression in China. Experimental AD was induced in rats by an intraperitoneal injection of d-galactose (d-gal) and administered intragastrically with aluminum chloride (AlCl3) simultaneously for 105 days. Morris water maze task as a behavior test was used for testing the effects of KXS on AD model and pathological changes to the brain were assessed by hematoxylin-eosin staining and immunohistochemistry. The levels of Bcl-2 and ChAT in hippocampus were evaluated by western-blot. Furthermore, metabolite profiling of AD was performed through ultra-performance liquid chromatography/electrospray ionization quadruple time-of- flight-high-definition mass spectrometry (UPLC/ESI-Q-TOF/HDMS) combined with pattern recognition approaches and pathway analysis. d-gal and AlCl3-treated caused a decline in spatial learning and memory, hippocampal histopathological abnormalities and increased Aβ1-40 levels in the brain cortex and hippocampus along with decreased Bcl-2 and ChAT expression in the hippocampus. KXS significantly improved the cognitive impairment induced by d-gal and AlCl3, attenuated hippocampal histopathological abnormalities, reduced Aβ1-40 levels and increased Bcl-2 and ChAT expression in the hippocampus. A total of 48 metabolites were considered as potential biomarkers of AD, and 36 metabolites may correlate with the regulation of KXS treatment on AD. Changes in AD metabolic profiling were close to normal states through regulating multiple perturbed pathways after KXS treatment. This study has revealed the potential biomarkers and metabolic networks of AD, illuminated the biochemistry

  10. Kai-xin-san, a chinese herbal decoction containing ginseng radix et rhizoma, polygalae radix, acori tatarinowii rhizoma, and poria, stimulates the expression and secretion of neurotrophic factors in cultured astrocytes.

    PubMed

    Zhu, Kevin Yue; Xu, Sherry Li; Choi, Roy Chi-Yan; Yan, Artemis Lu; Dong, Tina Ting-Xia; Tsim, Karl Wah-Keung

    2013-01-01

    Kai-xin-san (KXS), a Chinese herbal decoction prescribed by Sun Simiao in Beiji Qianjin Yaofang about 1400 years ago, contains Ginseng Radix et Rhizoma, Polygalae Radix, Acori Tatarinowii Rhizoma, and Poria. In China, KXS has been used to treat stress-related psychiatric diseases with the symptoms of depression and forgetfulness. Although animal study has supported the antidepression function of KXS, the mechanism in cellular level is still unknown. Here, a chemically standardized water extract of KXS was applied onto cultured astrocytes in exploring the action mechanisms of KXS treatment, which significantly stimulated the expression and secretion of neurotrophic factors, including NGF, BDNF, and GDNF, in a dose-dependent manner: the stimulation was both in mRNA and protein levels. In addition, the water extracts of four individual herbs did not significantly stimulate the expression of neurotrophic factors, which could explain the optimized effect of KXS in a herbal decoction. The KXS-induced expression of neurotrophic factors did not depend on signaling mediated by estrogen receptor or protein kinase. The results suggested that the antidepressant-like action of KXS might be mediated by an increase of expression of neurotrophic factors in astrocytes, which fully supported the clinical usage of this decoction. PMID:24222781

  11. Kai-Xin-San, a Chinese Herbal Decoction Containing Ginseng Radix et Rhizoma, Polygalae Radix, Acori Tatarinowii Rhizoma, and Poria, Stimulates the Expression and Secretion of Neurotrophic Factors in Cultured Astrocytes

    PubMed Central

    Zhu, Kevin Yue; Xu, Sherry Li; Choi, Roy Chi-Yan; Yan, Artemis Lu; Dong, Tina Ting-Xia; Tsim, Karl Wah-Keung

    2013-01-01

    Kai-xin-san (KXS), a Chinese herbal decoction prescribed by Sun Simiao in Beiji Qianjin Yaofang about 1400 years ago, contains Ginseng Radix et Rhizoma, Polygalae Radix, Acori Tatarinowii Rhizoma, and Poria. In China, KXS has been used to treat stress-related psychiatric diseases with the symptoms of depression and forgetfulness. Although animal study has supported the antidepression function of KXS, the mechanism in cellular level is still unknown. Here, a chemically standardized water extract of KXS was applied onto cultured astrocytes in exploring the action mechanisms of KXS treatment, which significantly stimulated the expression and secretion of neurotrophic factors, including NGF, BDNF, and GDNF, in a dose-dependent manner: the stimulation was both in mRNA and protein levels. In addition, the water extracts of four individual herbs did not significantly stimulate the expression of neurotrophic factors, which could explain the optimized effect of KXS in a herbal decoction. The KXS-induced expression of neurotrophic factors did not depend on signaling mediated by estrogen receptor or protein kinase. The results suggested that the antidepressant-like action of KXS might be mediated by an increase of expression of neurotrophic factors in astrocytes, which fully supported the clinical usage of this decoction. PMID:24222781

  12. Will Kai Become a Skinhead? Cultures of Hate in Germany and the New Europe.

    ERIC Educational Resources Information Center

    Lhotzky, Stephan

    2001-01-01

    The rise of hate crimes and development of right-wing extremism among adolescents in Europe will require a response involving a multitude of measures in different areas and various levels. Combating this development will necessitate cooperation of experts in different fields. Individuals and groups will need to contribute knowledge in a way that…

  13. It Started in Hawai'i Kai: Reminiscences of 43 Years (and Counting) of Collaboration and Friendship.

    PubMed

    Popper, Arthur N; Fay, Richard R

    2016-01-01

    This paper discusses the 43+ year collaboration of Arthur Popper and Richard Fay. Over these years, we have co-authored over 30 papers and 55 books. The collaboration benefits from a strong friendship that includes our spouses and children. By any measure, our collaboration must be seen as being successful. The basis for this success is, we think, twofold. First, we have very complementary and overlapping research interests. This has enabled us to tackle issues, whether in research or in planning meetings or books, from different perspectives. Second, a hallmark of our successful collaboration has been our deep and close friendship and the extension of that friendship to our spouses and children. In this paper, we discuss some of the events that have shaped our collaboration, and some of the people who have impacted our lives. PMID:26515309

  14. Intricate protein-protein interactions in the cyanobacterial circadian clock.

    PubMed

    Egli, Martin

    2014-08-01

    The cyanobacterial circadian clock consists of a post-translational oscillator (PTO) and a PTO-dependent transcription-translation feedback loop (TTFL). The PTO can be reconstituted in vitro with the KaiA, KaiB, and KaiC proteins, enabling detailed biochemical and biophysical investigations. Both the CI and the CII halves of the KaiC hexamer harbor ATPases, but only the C-terminal CII ring exhibits kinase and phospho-transferase activities. KaiA stimulates the kinase and KaiB associates with KaiC during the dephosphorylation phase and sequesters KaiA. Recent research has led to conflicting models of the KaiB-KaiC interaction, precluding a clear understanding of KaiB function and KaiABC clock mechanism. PMID:24936066

  15. Intricate Protein-Protein Interactions in the Cyanobacterial Circadian Clock*

    PubMed Central

    Egli, Martin

    2014-01-01

    The cyanobacterial circadian clock consists of a post-translational oscillator (PTO) and a PTO-dependent transcription-translation feedback loop (TTFL). The PTO can be reconstituted in vitro with the KaiA, KaiB, and KaiC proteins, enabling detailed biochemical and biophysical investigations. Both the CI and the CII halves of the KaiC hexamer harbor ATPases, but only the C-terminal CII ring exhibits kinase and phospho-transferase activities. KaiA stimulates the kinase and KaiB associates with KaiC during the dephosphorylation phase and sequesters KaiA. Recent research has led to conflicting models of the KaiB-KaiC interaction, precluding a clear understanding of KaiB function and KaiABC clock mechanism. PMID:24936066

  16. Combined SAXS/EM Based Models of the S. elongatus Post-Translational Circadian Oscillator and its Interactions with the Output His-Kinase SasA

    SciTech Connect

    Pattanayek, Rekha; Williams, Dewight R.; Rossi, Gian; Weigand, Steven; Mori, Tetsuya; Johnson, Carl H.; Stewart, Phoebe L.; Egli, Martin

    2012-03-15

    The circadian clock in the cyanobacterium Synechococcus elongatus is composed of a post-translational oscillator (PTO) that can be reconstituted in vitro from three different proteins in the presence of ATP and a transcription-translation feedback loop (TTFL). The homo-hexameric KaiC kinase, phosphatase and ATPase alternates between hypo- and hyper-phosphorylated states over the 24-h cycle, with KaiA enhancing phosphorylation, and KaiB antagonizing KaiA and promoting KaiC subunit exchange. SasA is a His kinase that relays output signals from the PTO formed by the three Kai proteins to the TTFL. Although the crystal structures for all three Kai proteins are known, atomic resolution structures of Kai and Kai/SasA protein complexes have remained elusive. Here, we present models of the KaiAC and KaiBC complexes derived from solution small angle X-ray scattering (SAXS), which are consistent with previous EM based models. We also present a combined SAXS/EM model of the KaiC/SasA complex, which has two N-terminal SasA sensory domains occupying positions on the C-terminal KaiC ring reminiscent of the orientations adopted by KaiB dimers. Using EM we demonstrate that KaiB and SasA compete for similar binding sites on KaiC. We also propose an EM based model of the ternary KaiABC complex that is consistent with the sequestering of KaiA by KaiB on KaiC during the PTO dephosphorylation phase. This work provides the first 3D-catalogue of protein-protein interactions in the KaiABC PTO and the output pathway mediated by SasA.

  17. Combined SAXS/EM based models of the S. elongatus post-translational circadian oscillator and its interactions with the output His-kinase SasA.

    PubMed

    Pattanayek, Rekha; Williams, Dewight R; Rossi, Gian; Weigand, Steven; Mori, Tetsuya; Johnson, Carl H; Stewart, Phoebe L; Egli, Martin

    2011-01-01

    The circadian clock in the cyanobacterium Synechococcus elongatus is composed of a post-translational oscillator (PTO) that can be reconstituted in vitro from three different proteins in the presence of ATP and a transcription-translation feedback loop (TTFL). The homo-hexameric KaiC kinase, phosphatase and ATPase alternates between hypo- and hyper-phosphorylated states over the 24-h cycle, with KaiA enhancing phosphorylation, and KaiB antagonizing KaiA and promoting KaiC subunit exchange. SasA is a His kinase that relays output signals from the PTO formed by the three Kai proteins to the TTFL. Although the crystal structures for all three Kai proteins are known, atomic resolution structures of Kai and Kai/SasA protein complexes have remained elusive. Here, we present models of the KaiAC and KaiBC complexes derived from solution small angle X-ray scattering (SAXS), which are consistent with previous EM based models. We also present a combined SAXS/EM model of the KaiC/SasA complex, which has two N-terminal SasA sensory domains occupying positions on the C-terminal KaiC ring reminiscent of the orientations adopted by KaiB dimers. Using EM we demonstrate that KaiB and SasA compete for similar binding sites on KaiC. We also propose an EM based model of the ternary KaiABC complex that is consistent with the sequestering of KaiA by KaiB on KaiC during the PTO dephosphorylation phase. This work provides the first 3D-catalogue of protein-protein interactions in the KaiABC PTO and the output pathway mediated by SasA. PMID:21887298

  18. Combined SAXS/EM Based Models of the S. elongatus Post-Translational Circadian Oscillator and its Interactions with the Output His-Kinase SasA

    PubMed Central

    Pattanayek, Rekha; Williams, Dewight R.; Rossi, Gian; Weigand, Steven; Mori, Tetsuya; Johnson, Carl H.; Stewart, Phoebe L.; Egli, Martin

    2011-01-01

    The circadian clock in the cyanobacterium Synechococcus elongatus is composed of a post-translational oscillator (PTO) that can be reconstituted in vitro from three different proteins in the presence of ATP and a transcription-translation feedback loop (TTFL). The homo-hexameric KaiC kinase, phosphatase and ATPase alternates between hypo- and hyper-phosphorylated states over the 24-h cycle, with KaiA enhancing phosphorylation, and KaiB antagonizing KaiA and promoting KaiC subunit exchange. SasA is a His kinase that relays output signals from the PTO formed by the three Kai proteins to the TTFL. Although the crystal structures for all three Kai proteins are known, atomic resolution structures of Kai and Kai/SasA protein complexes have remained elusive. Here, we present models of the KaiAC and KaiBC complexes derived from solution small angle X-ray scattering (SAXS), which are consistent with previous EM based models. We also present a combined SAXS/EM model of the KaiC/SasA complex, which has two N-terminal SasA sensory domains occupying positions on the C-terminal KaiC ring reminiscent of the orientations adopted by KaiB dimers. Using EM we demonstrate that KaiB and SasA compete for similar binding sites on KaiC. We also propose an EM based model of the ternary KaiABC complex that is consistent with the sequestering of KaiA by KaiB on KaiC during the PTO dephosphorylation phase. This work provides the first 3D-catalogue of protein-protein interactions in the KaiABC PTO and the output pathway mediated by SasA. PMID:21887298

  19. Keith Attenborough, Kai Ming Li and Kirill Horoshenkov, Predicting Outdoor Sound , Taylor & Francis, UK (2006) ISBN 9780419235101 xiii+441pp., price £75.

    NASA Astrophysics Data System (ADS)

    Gilbert, Kenneth E.

    2008-08-01

    In the preface to "Predicting Outdoor Sound," the authors say they regard their book as complementary to the book, "Computational Acoustics," by Erik M. Salomons. Further, they say that workers in outdoor sound would be wise to own both books. With these statements in mind, it seems prudent to briefly compare the two books. Put simply, "Computational Acoustics" is mainly about using full-wave, numerical models to predict long-range sound propagation in a realistic atmosphere over irregular, but otherwise simple, terrain. In contrast, "Predicting Outdoor Sound" is mainly about how to predict sound propagation in a homogeneous atmosphere over surfaces that range from simple flat impedance planes to very complex structures such as realistic barriers and buildings. There are other important differences as well. While "Computational Acoustics" is largely theoretical, "Predicting Outdoor Sound" has a wealth of experimental measurements and empirical prediction schemes. Specifically, Chapters 1-6 contain in-depth discussions of propagation in a homogeneous atmosphere over a flat, finite impedance surface. A multitude of ground impedance measurements and models are discussed in these chapters. Chapter 7 discusses the effect of source motion. Chapter 8 considers sound propagation over a flat surface with a horizontally varying impedance. Chapters 9 and 10 discuss, respectively, the performance of outdoor noise barriers and the effects of vegetation, trees and turbulence. Chapter 11 considers approximate analytic models for sound propagation that include the effects of a finite impedance surface, refraction, and turbulence. In Chapter 12, a number of currently popular, semi-empirical noise prediction schemes are discussed. Chapter 13, the final chapter, discusses sound and noise in an urban environment, with consideration of such issues as street canyons and building facades. Although the book probably would not serve well as a stand-alone text, the authors have nevertheless succeeded admirably in fulfilling their purpose: "…to provide a comprehensive reference about aspects of outdoor sound and its prediction that should be useful to practitioners, and yet is respectable from the academic point of view." As a complete and authoritative source of information on the effects of surfaces of all kinds (ground impedance, barriers, buildings, etc.) on sound propagation, "Predicting Outdoor Sound" is unique and indispensable.

  20. Rhythmic ring–ring stacking drives the circadian oscillator clockwise

    PubMed Central

    Chang, Yong-Gang; Tseng, Roger; Kuo, Nai-Wei; LiWang, Andy

    2012-01-01

    The oscillator of the circadian clock of cyanobacteria is composed of three proteins, KaiA, KaiB, and KaiC, which together generate a self-sustained ∼24-h rhythm of phosphorylation of KaiC. The mechanism propelling this oscillator has remained elusive, however. We show that stacking interactions between the CI and CII rings of KaiC drive the transition from the phosphorylation-specific KaiC–KaiA interaction to the dephosphorylation-specific KaiC–KaiB interaction. We have identified the KaiB-binding site, which is on the CI domain. This site is hidden when CI domains are associated as a hexameric ring. However, stacking of the CI and CII rings exposes the KaiB-binding site. Because the clock output protein SasA also binds to CI and competes with KaiB for binding, ring stacking likely regulates clock output. We demonstrate that ADP can expose the KaiB-binding site in the absence of ring stacking, providing an explanation for how it can reset the clock. PMID:22967510

  1. A Protein Fold Switch Joins the Circadian Oscillator to Clock Output in Cyanobacteria

    PubMed Central

    Chang, Yong-Gang; Cohen, Susan E.; Phong, Connie; Myers, William K.; Kim, Yong-Ick; Tseng, Roger; Lin, Jenny; Zhang, Li; Boyd, Joseph S.; Lee, Yvonne; Kang, Shannon; Lee, David; Li, Sheng; Britt, R. David; Rust, Michael J.; Golden, Susan S.; LiWang, Andy

    2015-01-01

    Organisms are adapted to the relentless cycles of day and night, because they evolved timekeeping systems called circadian clocks, which regulate biological activities with ~24-h rhythms. The clock of cyanobacteria is driven by a three-protein oscillator comprised of KaiA, KaiB, and KaiC, which together generate a circadian rhythm of KaiC phosphorylation. We show that KaiB flips between two distinct three-dimensional folds, and its rare transition to an active state provides a time delay that is required to match the timing of the oscillator to that of earth’s rotation. Once KaiB switches folds, it binds phosphorylated KaiC and captures KaiA, initiating a phase transition of the circadian cycle, and regulates components of the clock-output pathway, providing the link that joins the timekeeping and signaling functions of the oscillator. PMID:26113641

  2. Circadian rhythms. A protein fold switch joins the circadian oscillator to clock output in cyanobacteria.

    PubMed

    Chang, Yong-Gang; Cohen, Susan E; Phong, Connie; Myers, William K; Kim, Yong-Ick; Tseng, Roger; Lin, Jenny; Zhang, Li; Boyd, Joseph S; Lee, Yvonne; Kang, Shannon; Lee, David; Li, Sheng; Britt, R David; Rust, Michael J; Golden, Susan S; LiWang, Andy

    2015-07-17

    Organisms are adapted to the relentless cycles of day and night, because they evolved timekeeping systems called circadian clocks, which regulate biological activities with ~24-hour rhythms. The clock of cyanobacteria is driven by a three-protein oscillator composed of KaiA, KaiB, and KaiC, which together generate a circadian rhythm of KaiC phosphorylation. We show that KaiB flips between two distinct three-dimensional folds, and its rare transition to an active state provides a time delay that is required to match the timing of the oscillator to that of Earth's rotation. Once KaiB switches folds, it binds phosphorylated KaiC and captures KaiA, which initiates a phase transition of the circadian cycle, and it regulates components of the clock-output pathway, which provides the link that joins the timekeeping and signaling functions of the oscillator. PMID:26113641

  3. Mixtures of opposing phosphorylations within hexamers precisely time feedback in the cyanobacterial circadian clock

    PubMed Central

    Lin, Jenny; Chew, Justin; Chockanathan, Udaysankar; Rust, Michael J.

    2014-01-01

    Circadian oscillations are generated by the purified cyanobacterial clock proteins, KaiA, KaiB, and KaiC, through rhythmic interactions that depend on multisite phosphorylation of KaiC. However, the mechanisms that allow these phosphorylation reactions to robustly control the timing of oscillations over a range of protein stoichiometries are not clear. We show that when KaiC hexamers consist of a mixture of differentially phosphorylated subunits, the two phosphorylation sites have opposing effects on the ability of each hexamer to bind to the negative regulator KaiB. We likewise show that the ability of the positive regulator KaiA to act on KaiC depends on the phosphorylation state of the hexamer and that KaiA and KaiB recognize alternative allosteric states of the KaiC ring. Using mathematical models with kinetic parameters taken from experimental data, we find that antagonism of the two KaiC phosphorylation sites generates an ultrasensitive switch in negative feedback strength necessary for stable circadian oscillations over a range of component concentrations. Similar strategies based on opposing modifications may be used to support robustness in other timing systems and in cellular signaling more generally. PMID:25197081

  4. Dynamic Localization of the Cyanobacterial Circadian Clock Proteins

    PubMed Central

    Cohen, Susan E.; Erb, Marcella L.; Selimkhanov, Jangir; Dong, Guogang; Hasty, Jeff; Pogliano, Joe; Golden, Susan S.

    2014-01-01

    SUMMARY Background The cyanobacterial circadian clock system has been extensively studied and the structures, interactions and biochemical activities of the central oscillator proteins (KaiA, KaiB and KaiC) have been well elucidated. Despite this rich repository of information, little is known about the distribution of these proteins within the cell. Results Here we report that KaiA and KaiC localize as discrete foci near a single pole of cells in a clock-dependent fashion, with enhanced polar localization observed at night. KaiA localization is dependent on KaiC; consistent with this notion, KaiA and KaiC co-localize with each other as well as with CikA, a key input/output factor previously reported to display unipolar localization. The molecular mechanism that localizes KaiC to the poles is conserved in Escherichia coli, another Gram-negative rod shaped bacterium, suggesting that KaiC localization is not dependent on other clock- or cyanobacterial-specific factors. Moreover, expression of CikA mutant variants that distribute diffusely results in the striking de-localization of KaiC. Conclusions This work shows that the cyanobacterial circadian system undergoes a circadian orchestration of subcellular organization. We propose that the observed spatiotemporal localization pattern represents a novel layer of regulation that contributes to the robustness of the clock by facilitating protein complex formation and synchronizing the clock with environmental stimuli. PMID:25127213

  5. Adaptors and Innovators: A Description and Measure

    ERIC Educational Resources Information Center

    Kirton, Michael

    1976-01-01

    Describes development of the Kirton Adaption Innovation Inventory (KAI) for rating respondents on a continuum of adaptiveness-innovativeness, discusses tests of the validity and utility of the KAI model, and evaluates the KAI model's characteristics. For availability see EA 507 670. (Author/JG)

  6. Revealing a Two-Loop Transcriptional Feedback Mechanism in the Cyanobacterial Circadian Clock

    PubMed Central

    Hertel, Stefanie; Brettschneider, Christian; Axmann, Ilka M.

    2013-01-01

    Molecular genetic studies in the circadian model organism Synechococcus have revealed that the KaiC protein, the central component of the circadian clock in cyanobacteria, is involved in activation and repression of its own gene transcription. During 24 hours, KaiC hexamers run through different phospho-states during daytime. So far, it has remained unclear which phospho-state of KaiC promotes kaiBC expression and which opposes transcriptional activation. We systematically analyzed various combinations of positive and negative transcriptional feedback regulation by introducing a combined TTFL/PTO model consisting of our previous post-translational oscillator that considers all four phospho-states of KaiC and a transcriptional/translational feedback loop. Only a particular two-loop feedback mechanism out of 32 we have extensively tested is able to reproduce existing experimental observations, including the effects of knockout or overexpression of kai genes. Here, threonine and double phosphorylated KaiC hexamers activate and unphosphorylated KaiC hexamers suppress kaiBC transcription. Our model simulations suggest that the peak expression ratio of the positive and the negative component of kaiBC expression is the main factor for how the different two-loop feedback models respond to removal or to overexpression of kai genes. We discuss parallels between our proposed TTFL/PTO model and two-loop feedback structures found in the mammalian clock. PMID:23516349

  7. Structural and dynamic aspects of protein clocks: how can they be so slow and stable?

    PubMed

    Akiyama, Shuji

    2012-07-01

    KaiC is a core protein of the cyanobacterial Kai oscillator, which persists without transcription-translation feedback. In the presence of KaiA and KaiB, KaiC reveals rhythmic activation/inactivation of its ATPase and autokinase/autophosphotase activities over approximately 24 h. Since the in vitro reconstruction of the Kai oscillator, the structures and functions of the Kai proteins have been studied extensively. Each protein's crystal structure and low-resolution views of Kai complexes have been reported. In addition, newer data are emerging on dynamic aspects such as assembly/disassembly of the Kai components and a ticking motion of KaiC, which is probably coupled to its slow, temperature-compensated ATPase activity. The accumulated evidence offers an ideal opportunity to revisit a fundamental question regarding biological circadian clocks: what determines the temperature-compensated 24 h period? In this review, I summarize the current understanding of the Kai oscillator's molecular mechanism and discuss emerging ideas on protein clocks. PMID:22273739

  8. How a cyanobacterium tells time.

    PubMed

    Dong, Guogang; Golden, Susan S

    2008-12-01

    The cyanobacterium Synechococcus elongatus builds a circadian clock on an oscillator composed of three proteins, KaiA, KaiB, and KaiC, which can recapitulate a circadian rhythm of KaiC phosphorylation in vitro. The molecular structures of all three proteins are known, and the phosphorylation steps of KaiC, the interaction dynamics among the three Kai proteins, and a weak ATPase activity of KaiC have all been characterized. An input pathway of redox-sensitive proteins uses photosynthetic function to relay light/dark information to the oscillator, and signal transduction proteins of well-known families broadcast temporal information to the genome, where global changes in transcription and a compaction of the chromosome are clock regulated. PMID:18983934

  9. A Selaginella moellendorffii Ortholog of KARRIKIN INSENSITIVE2 Functions in Arabidopsis Development but Cannot Mediate Responses to Karrikins or Strigolactones[OPEN

    PubMed Central

    Waters, Mark T.; Scaffidi, Adrian; Moulin, Solène L.Y.; Sun, Yueming K.; Flematti, Gavin R.; Smith, Steven M.

    2015-01-01

    In Arabidopsis thaliana, the α/β-fold hydrolase KARRIKIN INSENSITIVE2 (KAI2) is essential for normal seed germination, seedling development, and leaf morphogenesis, as well as for responses to karrikins. KAI2 is a paralog of DWARF14 (D14), the proposed strigolactone receptor, but the evolutionary timing of functional divergence between the KAI2 and D14 clades has not been established. By swapping gene promoters, we show that Arabidopsis KAI2 and D14 proteins are functionally distinct. We show that the catalytic serine of KAI2 is essential for function in plants and for biochemical activity in vitro. We identified two KAI2 homologs from Selaginella moellendorffii and two from Marchantia polymorpha. One from each species could hydrolyze the strigolactone analog GR24 in vitro, but when tested for their ability to complement Arabidopsis d14 and kai2 mutants, neither of these homologs was effective. However, the second KAI2 homolog from S. moellendorffii was able to complement the seedling and leaf development phenotypes of Arabidopsis kai2. This homolog could not transduce signals from exogenous karrikins, strigolactone analogs, or carlactone, but its activity did depend on the conserved catalytic serine. We conclude that KAI2, and most likely the endogenous signal to which it responds, has been conserved since the divergence of lycophytes and angiosperm lineages, despite their major developmental and morphogenic differences. PMID:26175507

  10. Comment on: "Crustal strength in central Tibet determined from Holocene shoreline deflection around Siling Co" by Xuhua Shi, Eric Kirby, Kevin P. Furlong, Kai Meng, Ruth Robinson and Erchie Wang

    NASA Astrophysics Data System (ADS)

    England, Philip C.; Walker, Richard T.

    2016-01-01

    Shi et al. (2015) analysed the distributions of elevation of palaeoshorelines around Siling Tso, central Tibet assuming a model in which surface loads are entirely supported by an elastic lid overlying an inviscid fluid. They concluded that the thickness of the elastic lid is 20-30 km and that, for the assumption of an inviscid substrate in this model to be valid, the viscosity of the crust below the elastic lid must be less than 1- 2 ×1019 Pas. Here we relax the assumption of an inviscid lower crust and show that the distribution of shoreline elevations may be explained either by a thick elastic lid or by high viscosity in the lower crust. In the limit of an inviscid lower crust, the thickness of the elastic lid must be greater than 25 to 39 km. If the elastic lid is thinner than this, then the viscosity of the crust beneath must be at least 5 ×1019 to 2 ×1020 Pas, depending on the time interval over which the lake was loaded.

  11. Distinguishing Feedback Mechanisms in Clock Models

    NASA Astrophysics Data System (ADS)

    Golden, Alexander; Lubensky, David

    Biological oscillators are very diverse but can be classified based on dynamical motifs such as type of feedback. The S. Elongatus circadian oscillator is a novel circadian oscillator that can operate at constant protein number by modifying covalent states. It can be reproduced in vitro with only 3 different purified proteins: KaiA, KaiB, and KaiC. We use computational and analytic techniques to compare models of the S. Elongatus post-translational oscillator that rely on positive feedback with models that rely on negative feedback. We show that introducing a protein that binds competitively with KaiA to the KaiB-KaiC complex can distinguish between positive and negative feedback as the primary driver of the rhythm, which has so far been difficult to address experimentally. NSF Grant DMR-1056456.

  12. Structural and biophysical methods to analyze clock function and mechanism.

    PubMed

    Egli, Martin

    2015-01-01

    Structural approaches have provided insight into the mechanisms of circadian clock oscillators. This review focuses upon the myriad structural methods that have been applied to the molecular architecture of cyanobacterial circadian proteins, their interactions with each other, and the mechanism of the KaiABC posttranslational oscillator. X-ray crystallography and solution NMR were deployed to gain an understanding of the three-dimensional structures of the three proteins KaiA, KaiB, and KaiC that make up the inner timer in cyanobacteria. A hybrid structural biology approach including crystallography, electron microscopy, and solution scattering has shed light on the shapes of binary and ternary Kai protein complexes. Structural studies of the cyanobacterial oscillator demonstrate both the strengths and the limitations of the divide-and-conquer strategy. Thus, investigations of complexes involving domains and/or peptides have afforded valuable information into Kai protein interactions. However, high-resolution structural data are still needed at the level of complexes between the 360-kDa KaiC hexamer that forms the heart of the clock and its KaiA and KaiB partners. PMID:25662460

  13. Circadian transcriptional regulation by the posttranslational oscillator without de novo clock gene expression in Synechococcus

    PubMed Central

    Hosokawa, Norimune; Hatakeyama, Tetsuhiro S.; Kojima, Takashi; Kikuchi, Yoshiyuki; Ito, Hiroshi; Iwasaki, Hideo

    2011-01-01

    Circadian rhythms are a fundamental property of most organisms, from cyanobacteria to humans. In the unicellular obligately photoautotrophic cyanobacterium Synechococcus elongatus PCC 7942, essentially all promoter activities are controlled by the KaiABC-based clock under continuous light conditions. When Synechococcus cells are transferred from the light to continuous dark (DD) conditions, the expression of most genes, including the clock genes kaiA and kaiBC, is rapidly down-regulated, whereas the KaiC phosphorylation cycle persists. Therefore, we speculated that the posttranslational oscillator might not drive the transcriptional circadian output without de novo expression of the kai genes. Here we show that the cyanobacterial clock regulates the transcriptional output even in the dark. The expression of a subset of genes in the genomes of cells grown in the dark was dramatically affected by kaiABC nullification, and the magnitude of dark induction was dependent on the time at which the cells were transferred from the light to the dark. Moreover, under DD conditions, the expression of some dark-induced gene transcripts exhibited temperature-compensated damped oscillations, which were nullified in kaiABC-null strains and were affected by a kaiC period mutation. These results indicate that the Kai protein-based posttranslational oscillator can drive the circadian transcriptional output even without the de novo expression of the clock genes. PMID:21896749

  14. Daily Expression Pattern of Protein-Encoding Genes and Small Noncoding RNAs in Synechocystis sp. Strain PCC 6803

    PubMed Central

    Beck, Christian; Hertel, Stefanie; Rediger, Anne; Lehmann, Robert; Wiegard, Anika; Kölsch, Adrian; Heilmann, Beate; Georg, Jens; Hess, Wolfgang R.

    2014-01-01

    Many organisms harbor circadian clocks with periods close to 24 h. These cellular clocks allow organisms to anticipate the environmental cycles of day and night by synchronizing circadian rhythms with the rising and setting of the sun. These rhythms originate from the oscillator components of circadian clocks and control global gene expression and various cellular processes. The oscillator of photosynthetic cyanobacteria is composed of three proteins, KaiA, KaiB, and KaiC, linked to a complex regulatory network. Synechocystis sp. strain PCC 6803 possesses the standard cyanobacterial kaiABC gene cluster plus multiple kaiB and kaiC gene copies and antisense RNAs for almost every kai transcript. However, there is no clear evidence of circadian rhythms in Synechocystis sp. PCC 6803 under various experimental conditions. It is also still unknown if and to what extent the multiple kai gene copies and kai antisense RNAs affect circadian timing. Moreover, a large number of small noncoding RNAs whose accumulation dynamics over time have not yet been monitored are known for Synechocystis sp. PCC 6803. Here we performed a 48-h time series transcriptome analysis of Synechocystis sp. PCC 6803, taking into account periodic light-dark phases, continuous light, and continuous darkness. We found that expression of functionally related genes occurred in different phases of day and night. Moreover, we found day-peaking and night-peaking transcripts among the small RNAs; in particular, the amounts of kai antisense RNAs correlated or anticorrelated with those of their respective kai target mRNAs, pointing toward the regulatory relevance of these antisense RNAs. Surprisingly, we observed that the amounts of 16S and 23S rRNAs in this cyanobacterium fluctuated in light-dark periods, showing maximum accumulation in the dark phase. Importantly, the amounts of all transcripts, including small noncoding RNAs, did not show any rhythm under continuous light or darkness, indicating the absence

  15. 75 FR 3963 - Requested Administrative Waiver of the Coastwise Trade Laws

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-01-25

    ....S.C. 12121 and MARAD's regulations at 46 CFR part 388 (68 FR 23084; April 30, 2003), that the... administrative waiver of the Coastwise Trade Laws for the vessel MELE KAI. SUMMARY: As authorized by 46 U.S.C... vessel MELE KAI is: Intended Commercial Use of Vessel: ``Sport fishing out of Nawiliwili Harbor and...

  16. Two antagonistic clock-regulated histidine kinases time the activation of circadian gene expression

    PubMed Central

    Gutu, Andrian; O’Shea, Erin K.

    2013-01-01

    Summary The cyanobacterial circadian pacemaker consists of a three-protein clock – KaiA, KaiB and KaiC – that generates oscillations in the phosphorylation state of KaiC. Here we investigate how temporal information encoded in KaiC phosphorylation is transduced to RpaA, a transcription factor required for circadian gene expression. We show that phosphorylation of RpaA is regulated by two antagonistic histidine kinases, SasA and CikA, which are sequentially activated at distinct times by the Kai clock complex. SasA acts as a kinase toward RpaA, whereas CikA, previously implicated in clock input, acts as a phosphatase that dephosphorylates RpaA. CikA and SasA cooperate to generate an oscillation of RpaA activity that is distinct from that generated by either enzyme alone and offset from the rhythm of KaiC phosphorylation. Our observations reveal how circadian clocks can precisely control the timing of output pathways via the concerted action of two oppositely acting enzymes. PMID:23541768

  17. An allele of the crm gene blocks cyanobacterial circadian rhythms

    PubMed Central

    Boyd, Joseph S.; Bordowitz, Juliana R.; Bree, Anna C.; Golden, Susan S.

    2013-01-01

    The SasA-RpaA two-component system constitutes a key output pathway of the cyanobacterial Kai circadian oscillator. To date, rhythm of phycobilisome associated (rpaA) is the only gene other than kaiA, kaiB, and kaiC, which encode the oscillator itself, whose mutation causes completely arrhythmic gene expression. Here we report a unique transposon insertion allele in a small ORF located immediately upstream of rpaA in Synechococcus elongatus PCC 7942 termed crm (for circadian rhythmicity modulator), which results in arrhythmic promoter activity but does not affect steady-state levels of RpaA. The crm ORF complements the defect when expressed in trans, but only if it can be translated, suggesting that crm encodes a small protein. The crm1 insertion allele phenotypes are distinct from those of an rpaA null; crm1 mutants are able to grow in a light:dark cycle and have no detectable oscillations of KaiC phosphorylation, whereas low-amplitude KaiC phosphorylation rhythms persist in the absence of RpaA. Levels of phosphorylated RpaA in vivo measured over time are significantly altered compared with WT in the crm1 mutant as well as in the absence of KaiC. Taken together, these results are consistent with the hypothesis that the Crm polypeptide modulates a circadian-specific activity of RpaA. PMID:23918383

  18. A circadian clock nanomachine that runs without transcription or translation

    PubMed Central

    Egli, Martin; Johnson, Carl Hirschie

    2013-01-01

    The biochemical basis of circadian timekeeping is best characterized in cyanobacteria. The structures of its key molecular players, KaiA, KaiB, and KaiC are known and these proteins can reconstitute a remarkable circadian oscillation in a test tube. KaiC is rhythmically phosphorylated and its phospho-status is a marker of circadian phase that regulates ATPase activity and the oscillating assembly of a nanomachine. Analyses of the nanomachines have revealed how their timing circuit is ratcheted to be unidirectional and how they stay in synch to ensure a robust oscillator. These insights are likely to elucidate circadian timekeeping in higher organisms, including how transcription and translation could appear to be a core circadian timer when the true pacemaker is an embedded biochemical oscillator. PMID:23571120

  19. A circadian clock nanomachine that runs without transcription or translation.

    PubMed

    Egli, Martin; Johnson, Carl Hirschie

    2013-10-01

    The biochemical basis of circadian timekeeping is best characterized in cyanobacteria. The structures of its key molecular players, KaiA, KaiB, and KaiC are known and these proteins can reconstitute a remarkable circadian oscillation in a test tube. KaiC is rhythmically phosphorylated and its phospho-status is a marker of circadian phase that regulates ATPase activity and the oscillating assembly of a nanomachine. Analyses of the nanomachines have revealed how their timing circuit is ratcheted to be unidirectional and how they stay in synch to ensure a robust oscillator. These insights are likely to elucidate circadian timekeeping in higher organisms, including how transcription and translation could appear to be a core circadian timer when the true pacemaker is an embedded biochemical oscillator. PMID:23571120

  20. Kenya AIDS Indicator Surveys 2007 and 2012: Implications for Public Health Policies for HIV Prevention and Treatment

    PubMed Central

    Maina, William K.; Kim, Andrea A.; Rutherford, George W.; Harper, Malayah; K’Oyugi, Boniface O.; Sharif, Shahnaaz; Kichamu, George; Muraguri, Nicholas M.; Akhwale, Willis; De Cock, Kevin M.

    2016-01-01

    AIDS Indicator Surveys are standardized surveillance tools used by countries with generalized HIV epidemics to provide, in a timely fashion, indicators for effective monitoring of HIV. Such data should guide responses to the HIV epidemic, meet program reporting requirements, and ensure comparability of findings across countries and over time. Kenya has conducted 2 AIDS Indicator Surveys, in 2007 (KAIS 2007) and 2012–2013 (KAIS 2012). These nationally representative surveys have provided essential epidemiologic, socio-demographic, behavioral, and biologic data on HIV and related indicators to evaluate the national HIV response and inform policies for prevention and treatment of the disease. We present a summary of findings from KAIS 2007 and KAIS 2012 and the impact that these data have had on changing HIV policies and practice. PMID:24732817

  1. Elucidation of the Role of Clp Protease Components in Circadian Rhythm by Genetic Deletion and Overexpression in Cyanobacteria

    PubMed Central

    Kitayama, Yohko; Kondo, Takao

    2013-01-01

    In the cyanobacterium Synechococcus elongatus PCC7942, KaiA, KaiB, and KaiC are essential elements of the circadian clock, and Kai-based oscillation is thought to be the basic circadian timing mechanism. The Kai-based oscillator coupled with transcription/translation feedback and other intercellular factors maintains the stability of the 24-hour period in vivo. In this study, we showed that disruption of the Clp protease family genes clpP1, clpP2, and clpX and the overexpression of clpP3 cause long-period phenotypes. There were no significant changes in the levels of the clock proteins in these mutants. The overexpression of clpX led to a decrease in kaiBC promoter activity, the disruption of the circadian rhythm, and eventually cell death. However, after the transient overexpression of clpX, the kaiBC gene expression rhythm recovered after a few days. The rhythm phase after recovery was almost the same as the phase before clpX overexpression. These results suggest that the core Kai-based oscillation was not affected by clpX overexpression. Moreover, we showed that the overexpression of clpX sequentially upregulated ribosomal protein subunit mRNA levels, followed by upregulation of other genes, including the clock genes. Additionally, we found that the disruption of clpX decreased the expression of the ribosomal protein subunits. Finally, we showed that the circadian period was prolonged following the addition of a translation inhibitor at a low concentration. These results suggest that translational efficiency affects the circadian period and that clpX participates in the control of translation efficiency by regulating the transcription of ribosomal protein genes. PMID:23913328

  2. The circadian oscillator in Synechococcus elongatus controls metabolite partitioning during diurnal growth

    PubMed Central

    Diamond, Spencer; Jun, Darae; Rubin, Benjamin E.; Golden, Susan S.

    2015-01-01

    Synechococcus elongatus PCC 7942 is a genetically tractable model cyanobacterium that has been engineered to produce industrially relevant biomolecules and is the best-studied model for a prokaryotic circadian clock. However, the organism is commonly grown in continuous light in the laboratory, and data on metabolic processes under diurnal conditions are lacking. Moreover, the influence of the circadian clock on diurnal metabolism has been investigated only briefly. Here, we demonstrate that the circadian oscillator influences rhythms of metabolism during diurnal growth, even though light–dark cycles can drive metabolic rhythms independently. Moreover, the phenotype associated with loss of the core oscillator protein, KaiC, is distinct from that caused by absence of the circadian output transcriptional regulator, RpaA (regulator of phycobilisome-associated A). Although RpaA activity is important for carbon degradation at night, KaiC is dispensable for those processes. Untargeted metabolomics analysis and glycogen kinetics suggest that functional KaiC is important for metabolite partitioning in the morning. Additionally, output from the oscillator functions to inhibit RpaA activity in the morning, and kaiC-null strains expressing a mutant KaiC phosphomimetic, KaiC-pST, in which the oscillator is locked in the most active output state, phenocopies a ΔrpaA strain. Inhibition of RpaA by the oscillator in the morning suppresses metabolic processes that normally are active at night, and kaiC-null strains show indications of oxidative pentose phosphate pathway activation as well as increased abundance of primary metabolites. Inhibitory clock output may serve to allow secondary metabolite biosynthesis in the morning, and some metabolites resulting from these processes may feed back to reinforce clock timing. PMID:25825710

  3. The Cyanobacterial Circadian System: From Biophysics to Bioevolution

    PubMed Central

    Stewart, Phoebe L.; Egli, Martin

    2011-01-01

    Recent studies have unveiled the molecular machinery responsible for the biological clock in cyanobacteria and found that it exerts pervasive control over cellular processes including global gene expression. Indeed, the entire chromosome undergoes daily cycles of topology/compaction! The circadian system comprises both a posttranslational oscillator (PTO) and a transcriptional/translational feedback loop (TTFL). The PTO can be reconstituted in vitro with three purified proteins (KaiA, KaiB, and KaiC) and ATP. These are the only circadian proteins for which high-resolution structures are available. Phase in this nanoclockwork has been associated with key phosphorylations of KaiC. Structural considerations illuminate the mechanism by which the KaiABC oscillator ratchets unidirectionally. Models of the complete in vivo system have important implications for our understanding of circadian clocks in higher organisms, including mammals. The conjunction of structural, biophysical, and biochemical approaches to this system has brought our understanding of the molecular mechanisms of biological timekeeping to an unprecedented level. PMID:21332358

  4. Does Access to Care Still Affect Health Care Utilization by Immigrants? Testing of an Empirical Explanatory Model of Health Care Utilization by Korean American Immigrants with High Blood Pressure

    PubMed Central

    Han, Hae-Ra; Lee, Jong-Eun; Kim, Ji-Yun; Kim, Kim B.; Ryu, Jai Poong; Kim, Miyong

    2015-01-01

    Despite well-known benefits of health care utilization for the effective management of chronic diseases, the underlying mechanism of understanding health care utilization in ethnic minority population has not been systematically explored. The purpose of this paper is to examine the predictive ability of a health care utilization model by analyzing the interplay between predisposing, enabling, and need factors. The sample consisted of hypertensive Korean American immigrants (KAIs) 40–64 years of age who participated in a self-help intervention for high blood pressure care (SHIP-HBP). Using structured questionnaires, data were collected from 445 KAIs at baseline and analyzed with path analysis. Insurance status and relevant medical history were not just strong direct effects but also carried the most total effect on the health care utilization of these patients. Life priorities, years of residence in the US and perceived income level exerted indirect effects through the participants’ insurance status. Our statistical analysis indicated a good fit for the proposed model (x2 = 28.4, P = 0.29; NFI = 0.91; CFI = 0.99; RMSEA = 0.02). Overall, the model explained 18% of the variance in health care utilization of hypertensive KAIs. These findings strongly support a need to improve access to health care for KAIs by introducing a variety of community resources and building sustainable community infrastructures. PMID:19649709

  5. Collaborative Research: Neutrinos and Nucleosynthesis in Hot and Dense Matter

    SciTech Connect

    Alford, Mark

    2015-05-31

    The Topical Collaboration funded one of Prof. Alford's graduate students, Jun (Sophia) Han, by providing 75% of her support. The work reported here was wholly or partly supported by the Topical Collaboration. Additional support, e.g. for postdoc Kai Schwenzer, came from Nuclear Theory grant #DE-FG02-05ER41375.

  6. Effects of Typographic Variables on Eye-Movement Measures in Reading Chinese from a Screen

    ERIC Educational Resources Information Center

    Yen, Nai-Shing; Tsai, Jie-Li; Chen, Pei-Ling; Lin, Hsuan-Yu; Chen, Arbee L. P.

    2011-01-01

    To investigate the most efficient way to represent text in reading Chinese on computer displays, three typographic variables, character size (41[feet] arc/24 pixels and 60[feet] arc/32 pixels), character spacing (1/4 and 1/8 character width) and font type (Kai and Ming), were manipulated. Results showed that the reading speed for Chinese…

  7. The Impact of an Assurance System on the Quality of Teaching and Learning--Using the Example of a University in Russia and One of the Universities in Germany

    ERIC Educational Resources Information Center

    Szymenderski, Peggy; Yagudina, Liliya; Burenkova, Olga

    2015-01-01

    In this paper we consider the question of how quality assurance can have a real, positive impact on the quality of teaching and learning at universities, considering the realities of different systems--the system of control and the system of quality culture--in using the example of two universities: the KNITU-KAI in Russia and the TU Dresden in…

  8. On a Quest for English

    ERIC Educational Resources Information Center

    Waters, John K.

    2007-01-01

    This article reports the potential of online role-playing games to be a powerful tool for English as a second language (ESL) learning. When Professor Edd Schneider and game designer Kai Zheng suggested to attendees gathered in San Francisco last spring for the annual Game Developers Conference that massively multiplayer online role-playing games,…

  9. Font Effects of Chinese Characters and Pseudo-Characters on the N400: Evidence for an Orthographic Processing View

    ERIC Educational Resources Information Center

    Lv, Caixia; Wang, Quanhong

    2012-01-01

    Event-related brain potentials (ERPs) were recorded during a Chinese character decision task to examine whether N400 amplitude is modulated by stimulus font. Results revealed large negative-going ERPs in an N400 time window of 300-500 ms to stimuli presented in degraded Xing Kai Ti (XKT) font compared with more intact Song Ti (ST) font regardless…

  10. Influence of Creative Style and Gender on Students' Achievement in Physics

    ERIC Educational Resources Information Center

    Mkpanang, John T.

    2016-01-01

    The research investigated the influence of creative style and gender on students' achievement in physics. The sample consisting one hundred (100) Senior Secondary II physics students, made up of 50 males and 50 females in Oruk Anam Local Government Area of Akwa Ibom State, Nigeria, were administered the Kirton Adaptor-Innovator Inventory (KAI),…

  11. Purification, crystallization and preliminary crystallographic analysis of RecA superfamily ATPase PH0284 from Pyrococcus horikoshii OT3

    SciTech Connect

    Bagautdinov, Bagautdin; Kunishima, Naoki

    2006-04-01

    RecA superfamily ATPase PH0284 from P. horikoshii OT3 was overexpressed, purified, crystallized and cocrystallized with ATP. Both crystal forms belong to the trigonal space group P3{sub 2}21 and diffract X-rays to 2.0 and 2.3 Å resolution, respectively. Circadian (daily) protein clocks are found in cyanobacteria, where a complex of the KaiA, KaiB and KaiC proteins generates circadian rhythms. The 28.09 kDa KaiC homologue PH0284 protein from Pyrococcus horikoshii OT3 was cloned and expressed and the purified protein was crystallized by the oil-microbatch method at 295 K. X-ray diffraction data from the crystal were collected to 2.0 Å resolution using synchrotron radiation at 100 K. The crystal belongs to the trigonal space group P3{sub 2}21, with unit-cell parameters a = b = 96.06, c = 298.90 Å. Assuming the presence of one hexamer in the asymmetric unit gives a V{sub M} value of 2.36 Å{sup 3} Da{sup −1} and a solvent content of 47.9%. A cocrystal with ATP was prepared and a diffraction data set was collected at 2.3 Å resolution.

  12. Purification, crystallization and preliminary crystallographic analysis of RecA superfamily ATPase PH0284 from Pyrococcus horikoshii OT3.

    PubMed

    Bagautdinov, Bagautdin; Kunishima, Naoki

    2006-04-01

    Circadian (daily) protein clocks are found in cyanobacteria, where a complex of the KaiA, KaiB and KaiC proteins generates circadian rhythms. The 28.09 kDa KaiC homologue PH0284 protein from Pyrococcus horikoshii OT3 was cloned and expressed and the purified protein was crystallized by the oil-microbatch method at 295 K. X-ray diffraction data from the crystal were collected to 2.0 angstroms resolution using synchrotron radiation at 100 K. The crystal belongs to the trigonal space group P3(2)21, with unit-cell parameters a = b = 96.06, c = 298.90 angstroms. Assuming the presence of one hexamer in the asymmetric unit gives a V(M) value of 2.36 angstroms3 Da(-1) and a solvent content of 47.9%. A cocrystal with ATP was prepared and a diffraction data set was collected at 2.3 angstroms resolution. PMID:16582499

  13. Purification, crystallization and preliminary crystallographic analysis of RecA superfamily ATPase PH0284 from Pyrococcus horikoshii OT3

    PubMed Central

    Bagautdinov, Bagautdin; Kunishima, Naoki

    2006-01-01

    Circadian (daily) protein clocks are found in cyanobacteria, where a complex of the KaiA, KaiB and KaiC proteins generates circadian rhythms. The 28.09 kDa KaiC homologue PH0284 protein from Pyrococcus horikoshii OT3 was cloned and expressed and the purified protein was crystallized by the oil-microbatch method at 295 K. X-ray diffraction data from the crystal were collected to 2.0 Å resolution using synchrotron radiation at 100 K. The crystal belongs to the trigonal space group P3221, with unit-cell parameters a = b = 96.06, c = 298.90 Å. Assuming the presence of one hexamer in the asymmetric unit gives a V M value of 2.36 Å3 Da−1 and a solvent content of 47.9%. A cocrystal with ATP was prepared and a diffraction data set was collected at 2.3 Å resolution. PMID:16582499

  14. ACUTE-TO-CHRONIC ESTIMATION (ACE V 2.0) WITH TIME-CONCENTRATION-EFFECT MODELS: USER MANUAL AND SOFTWARE

    EPA Science Inventory

    Ellersieck, Mark R., Amha Asfaw, Foster L. Mayer, Gary F. Krause, Kai Sun and Gunhee Lee. 2003. Acute-to-Chronic Estimation (ACE v2.0) with Time-Concentration-Effect Models: User Manual and Software. EPA/600/R-03/107. U.S. Environmental Protection Agency, National Health and Envi...

  15. ICCE/ICCAI 2000 Full & Short Papers (Methodologies).

    ERIC Educational Resources Information Center

    2000

    This document contains the full text of the following full and short papers on methodologies from ICCE/ICCAI 2000 (International Conference on Computers in Education/International Conference on Computer-Assisted Instruction): (1) "A Methodology for Learning Pattern Analysis from Web Logs by Interpreting Web Page Contents" (Chih-Kai Chang and…

  16. Chinese Lessons: Shanghai's Rise to the Top of the PISA League Tables

    ERIC Educational Resources Information Center

    Tucker, Marc S., Ed.

    2014-01-01

    Marc Tucker, President of the National Center on Education and the Economy, presents this compilation of interviews with top Chinese education leaders and international researchers exploring some of the policies and practices behind Shanghai's outstanding performance on PISA 2009 and PISA 2012. The interviews include perspectives from Kai-ming…

  17. Expression of human epileptic temporal lobe neurotransmitter receptors in Xenopus oocytes: An innovative approach to study epilepsy.

    PubMed

    Palma, Eleonora; Esposito, Vincenzo; Mileo, Anna Maria; Di Gennaro, Giancarlo; Quarato, Pierpaolo; Giangaspero, Felice; Scoppetta, Ciriaco; Onorati, Paolo; Trettel, Flavia; Miledi, Ricardo; Eusebi, Fabrizio

    2002-11-12

    Poly(A(+)) RNA was extracted from the temporal lobe (TL) of medically intractable epileptic patients which underwent surgical TL resection. Injection of this mRNA into Xenopus oocytes led to the expression of ionotropic receptors for gamma-aminobutyric acid (GABA), kainate (KAI) and alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA). Membrane currents elicited by GABA inverted polarity at -15 mV, close to the oocyte's chloride equilibrium potential, were inhibited by bicuculline, and were potentiated by pentobarbital and flunitrazepam. These basic characteristics were also displayed by GABA currents elicited in oocytes injected with mRNAs isolated from human TL glioma (TLG) or from mouse TL. However, the GABA receptors expressed by the epileptic TL mRNA exhibited some unusual properties, consisting in a rapid current run-down after repetitive GABA applications and a large EC(50) (125 microM). AMPA alone evoked very small or nil currents, whereas KAI induced larger currents. Nevertheless, upon cyclothiazide treatment, AMPA elicited substantial currents that, like the KAI currents, were inhibited by 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX). Furthermore, the glutamate receptor 5 (GluR5) agonist, ATPA, failed to evoke an obvious current although both RT-PCR and Western blot analyses showed GluR5 expression in the epileptic TL. Oocytes injected with mouse TL or human TLG mRNAs generated KAI and AMPA currents similar to those evoked in oocytes injected with epileptic TL mRNA but, in contrast to these, the mouse TL and human TLG oocytes were also responsive to ATPA. Our findings are in accord with the concept that both a depression of GABA inhibition and a dysfunction of the KAI-receptor system maintain a high neuronal excitability that results in epileptic seizures. PMID:12409614

  18. Expression of human epileptic temporal lobe neurotransmitter receptors in Xenopus oocytes: An innovative approach to study epilepsy

    PubMed Central

    Palma, Eleonora; Esposito, Vincenzo; Mileo, Anna Maria; Di Gennaro, Giancarlo; Quarato, Pierpaolo; Giangaspero, Felice; Scoppetta, Ciriaco; Onorati, Paolo; Trettel, Flavia; Miledi, Ricardo; Eusebi, Fabrizio

    2002-01-01

    Poly(A+) RNA was extracted from the temporal lobe (TL) of medically intractable epileptic patients which underwent surgical TL resection. Injection of this mRNA into Xenopus oocytes led to the expression of ionotropic receptors for γ-aminobutyric acid (GABA), kainate (KAI) and α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA). Membrane currents elicited by GABA inverted polarity at −15 mV, close to the oocyte's chloride equilibrium potential, were inhibited by bicuculline, and were potentiated by pentobarbital and flunitrazepam. These basic characteristics were also displayed by GABA currents elicited in oocytes injected with mRNAs isolated from human TL glioma (TLG) or from mouse TL. However, the GABA receptors expressed by the epileptic TL mRNA exhibited some unusual properties, consisting in a rapid current run-down after repetitive GABA applications and a large EC50 (125 μM). AMPA alone evoked very small or nil currents, whereas KAI induced larger currents. Nevertheless, upon cyclothiazide treatment, AMPA elicited substantial currents that, like the KAI currents, were inhibited by 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX). Furthermore, the glutamate receptor 5 (GluR5) agonist, ATPA, failed to evoke an obvious current although both RT-PCR and Western blot analyses showed GluR5 expression in the epileptic TL. Oocytes injected with mouse TL or human TLG mRNAs generated KAI and AMPA currents similar to those evoked in oocytes injected with epileptic TL mRNA but, in contrast to these, the mouse TL and human TLG oocytes were also responsive to ATPA. Our findings are in accord with the concept that both a depression of GABA inhibition and a dysfunction of the KAI-receptor system maintain a high neuronal excitability that results in epileptic seizures. PMID:12409614

  19. The extent of grain yield and plant growth enhancement by plant growth-promoting broad-spectrum Streptomyces sp. in chickpea.

    PubMed

    Gopalakrishnan, Subramaniam; Srinivas, Vadlamudi; Alekhya, Gottumukkala; Prakash, Bandikinda; Kudapa, Himabindu; Rathore, Abhishek; Varshney, Rajeev Kumar

    2015-01-01

    The physiological and molecular responses of five strains of Streptomyces sp. (CAI-17, CAI-68, CAI-78, KAI-26 and KAI-27), with their proven potential for charcoal rot disease control in sorghum and plant growth-promotion (PGP) in sorghum and rice, were studied to understand the mechanisms causing the beneficial effects. In this investigation, those five strains were evaluated for their PGP capabilities in chickpea in the 2012-13 and 2013-14 post-rainy seasons. All of the Streptomyces sp. strains exhibited enhanced nodule number, nodule weight, root weight and shoot weight at 30 days after sowing (DAS) and pod number, pod weight, leaf area, leaf weight and stem weight at 60 DAS in both seasons over the un-inoculated control. At crop maturity, the Streptomyces strains had enhanced stover yield, grain yield, total dry matter and seed number plant(-1) in both seasons over the un-inoculated control. In the rhizosphere, the Streptomyces sp. also significantly enhanced microbial biomass carbon, dehydrogenase activity, total nitrogen, available phosphorous and organic carbon in both seasons over the un-inoculated control. Of the five strains of Streptomyces sp., CAI-17, CAI-68 and CAI-78 were superior to KAI-26 and KAI-27 in terms of their effects on root and shoot development, nodule formation and crop productivity. Scanning electron microscopy (SEM) micrographs had revealed the success in colonization of the chickpea roots by all five strains. Quantitative real-time PCR (qRT-PCR) analysis of selected PGP genes of actinomycetes revealed the selective up-regulation of indole-3-acetic acid (IAA)-related and siderophore-related genes by CAI-68 and of β-1,3-glucanase genes by KAI-26. PMID:25646153

  20. The Cyanobacterial Clock and Metabolism

    PubMed Central

    Pattanayak, Gopal; Rust, Michael J.

    2014-01-01

    Cyanobacteria possess the simplest known circadian clock, which presents a unique opportunity to study how rhythms are generated and how input signals from the environment reset the clock time. The kaiABC locus forms the core of the oscillator, and the remarkable ability to reconstitute oscillations using purified KaiABC proteins has allowed researchers to study mechanism using the tools of quantitative biochemistry. Autotrophic cyanobacteria experience major shifts in metabolism following a light-dark transition, and recent work suggests that input mechanisms that couple the day-night cycle to the clock involve energy and redox metabolites acting directly on clock proteins. We offer a summary of the current state of knowledge in this system and present a perspective for future lines of investigation. PMID:24667330

  1. Electrical control of spin in topological insulators

    NASA Astrophysics Data System (ADS)

    Chang, Kai

    2012-02-01

    by changing the gate voltage. It provides us a new way to control surface magnetism electrically. The gap opened by doped magnetic ions can lead to a short-range Bloembergen-Rowland interaction. The competition among the Heisenberg, Ising, and DM terms leads to rich spin configurations and an anomalous Hall effect on different lattices [4]. There are many proposals for quantum computation scheme are based on the spin in semiconductor quantum dots. Topological insulator quantum dots display a very different behavior with that of conventional semiconductor quantum dots [5]. In sharp contrast to conventional semiconductor quantum dots, the quantum states in the gap of the HgTe QD are fully spin-polarized and show ring-like density distributions near the boundary of the QD and optically dark. The persistent charge currents and magnetic moments, i.e., the Aharonov-Bohm effect, can be observed in such a QD structure. This feature offers us a practical way to detect these exotic ring-like edge states by using the SQUID technique. [0pt]Refs: [1] W. Yang, Kai Chang, and S. C. Zhang, Phys. Rev. Lett. 100, 056602 (2008); J. Li and Kai Chang, Appl. Phys. Lett. 95, 222110 (2009). [2] L. B. Zhang, Kai Chang, X. C. Xie, H. Buhmann and L. W. Molenkamp, New J. Phys. 12, 083058 (2010). [3] L. B. Zhang, F. Cheng, F. Zhai and Kai Chang, Phys. Rev. B 83 081402(R) (2011); Z. H. Wu, F. Zhai, F. M. Peeters, H. Q. Xu and Kai Chang, Phys, Rev. Lett. 106, 176802 (2011). [4] J. J. Zhu, D. X. Yao, S. C. Zhang, and Kai Chang, Phys. Rev. Lett. 106, 097201 (2011). [5] Kai Chang, and Wen-Kai Lou, Phys. Rev. Lett. 106, 206802 (2011).

  2. Circadian rhythms. Atomic-scale origins of slowness in the cyanobacterial circadian clock.

    PubMed

    Abe, Jun; Hiyama, Takuya B; Mukaiyama, Atsushi; Son, Seyoung; Mori, Toshifumi; Saito, Shinji; Osako, Masato; Wolanin, Julie; Yamashita, Eiki; Kondo, Takao; Akiyama, Shuji

    2015-07-17

    Circadian clocks generate slow and ordered cellular dynamics but consist of fast-moving bio-macromolecules; consequently, the origins of the overall slowness remain unclear. We identified the adenosine triphosphate (ATP) catalytic region [adenosine triphosphatase (ATPase)] in the amino-terminal half of the clock protein KaiC as the minimal pacemaker that controls the in vivo frequency of the cyanobacterial clock. Crystal structures of the ATPase revealed that the slowness of this ATPase arises from sequestration of a lytic water molecule in an unfavorable position and coupling of ATP hydrolysis to a peptide isomerization with high activation energy. The slow ATPase is coupled with another ATPase catalyzing autodephosphorylation in the carboxyl-terminal half of KaiC, yielding the circadian response frequency of intermolecular interactions with other clock-related proteins that influences the transcription and translation cycle. PMID:26113637

  3. Evaluation of Streptomyces strains isolated from herbal vermicompost for their plant growth-promotion traits in rice.

    PubMed

    Gopalakrishnan, Subramaniam; Vadlamudi, Srinivas; Bandikinda, Prakash; Sathya, Arumugam; Vijayabharathi, Rajendran; Rupela, Om; Kudapa, Himabindu; Katta, Krishnamohan; Varshney, Rajeev Kumar

    2014-01-20

    Six actinomycetes, CAI-13, CAI-85, CAI-93, CAI-140, CAI-155 and KAI-180, isolated from six different herbal vermi-composts were characterized for in vitro plant growth-promoting (PGP) properties and further evaluated in the field for PGP activity in rice. Of the six actinomycetes, CAI-13, CAI-85, CAI-93, CAI-140 and CAI-155 produced siderophores; CAI-13, CAI-93, CAI-155 and KAI-180 produced chitinase; CAI-13, CAI-140, CAI-155 and KAI-180 produced lipase; CAI-13, CAI-93, CAI-155 and KAI-180 produced protease; and CAI-13, CAI-85, CAI-140 and CAI-155 produced ß-1-3-glucanase whereas all the six actinomycetes produced cellulase, hydrocyanic acid and indole acetic acid (IAA). The actinomycetes were able to grow in NaCl concentrations of up to 8%, at pH values between 7 and 11, temperatures between 20 and 40 °C and compatible with fungicide bavistin at field application levels. In the rice field, the actinomycetes significantly enhanced tiller numbers, panicle numbers, filled grain numbers and weight, stover yield, grain yield, total dry matter, root length, volume and dry weight over the un-inoculated control. In the rhizosphere, the actinomycetes also significantly enhanced total nitrogen, available phosphorous, % organic carbon, microbial biomass carbon and nitrogen and dehydrogenase activity over the un-inoculated control. Sequences of 16S rDNA gene of the actinomycetes matched with different Streptomyces species in BLAST analysis. Of the six actinomycetes, CAI-85 and CAI-93 were found superior over other actinomycetes in terms of PGP properties, root development and crop productivity. qRT-PCR analysis on selected plant growth promoting genes of actinomycetes revealed the up-regulation of IAA genes only in CAI-85 and CAI-93. PMID:24113511

  4. The Kenya AIDS Indicator Survey 2012: Rationale, Methods, Description of Participants, and Response Rates

    PubMed Central

    Waruiru, Wanjiru; Kim, Andrea A.; Kimanga, Davies O.; Ng’ang’a, James; Schwarcz, Sandra; Kimondo, Lucy; Ng’ang’a, Anne; Umuro, Mamo; Mwangi, Mary; Ojwang’, James K.; Maina, William K.

    2016-01-01

    Background Cross-sectional population-based surveys are essential surveillance tools for tracking changes in HIV epidemics. In 2007, Kenya implemented the first AIDS Indicator Survey [Kenya AIDS Indicator Survey (KAIS) 2007)], a nationally representative, population-based survey that collected demographic and behavioral data and blood specimens from individuals aged 15–64 years. Kenya’s second AIDS Indicator Survey (KAIS 2012) was conducted to monitor changes in the epidemic, evaluate HIV prevention, care, and treatment initiatives, and plan for an efficient and effective response to the HIV epidemic. Methods KAIS 2012 was a cross-sectional 2-stage cluster sampling design, household-based HIV serologic survey that collected information on households as well as demographic and behavioral data from Kenyans aged 18 months to 64 years. Participants also provided blood samples for HIV serology and other related tests at the National HIV Reference Laboratory. Results Among 9300 households sampled, 9189 (98.8%) were eligible for the survey. Of the eligible households, 8035 (87.4%) completed household-level questionnaires. Of 16,383 eligible individuals aged 15–64 years and emancipated minors aged less than 15 years in these households, 13,720 (83.7%) completed interviews; 11,626 (84.7%) of the interviewees provided a blood specimen. Of 6302 eligible children aged 18 months to 14 years, 4340 (68.9%) provided a blood specimen. Of the 2094 eligible children aged 10–14 years, 1661 (79.3%) completed interviews. Conclusions KAIS 2012 provided representative data to inform a strategic response to the HIV epidemic in the country. PMID:24732819

  5. Biobutanol production in a Clostridium acetobutylicum biofilm reactor integrated with simultaneous product recovery by adsorption

    PubMed Central

    2014-01-01

    Background Clostridium acetobutylicum can propagate on fibrous matrices and form biofilms that have improved butanol tolerance and a high fermentation rate and can be repeatedly used. Previously, a novel macroporous resin, KA-I, was synthesized in our laboratory and was demonstrated to be a good adsorbent with high selectivity and capacity for butanol recovery from a model solution. Based on these results, we aimed to develop a process integrating a biofilm reactor with simultaneous product recovery using the KA-I resin to maximize the production efficiency of biobutanol. Results KA-I showed great affinity for butanol and butyrate and could selectively enhance acetoin production at the expense of acetone during the fermentation. The biofilm reactor exhibited high productivity with considerably low broth turbidity during repeated batch fermentations. By maintaining the butanol level above 6.5 g/L in the biofilm reactor, butyrate adsorption by the KA-I resin was effectively reduced. Co-adsorption of acetone by the resin improved the fermentation performance. By redox modulation with methyl viologen (MV), the butanol-acetone ratio and the total product yield increased. An equivalent solvent titer of 96.5 to 130.7 g/L was achieved with a productivity of 1.0 to 1.5 g · L-1 · h-1. The solvent concentration and productivity increased by 4 to 6-fold and 3 to 5-fold, respectively, compared to traditional batch fermentation using planktonic culture. Conclusions Compared to the conventional process, the integrated process dramatically improved the productivity and reduced the energy consumption as well as water usage in biobutanol production. While genetic engineering focuses on strain improvement to enhance butanol production, process development can fully exploit the productivity of a strain and maximize the production efficiency. PMID:24401161

  6. [Analysis on the part of local medicinal herbs in Guangyangzaji (Guang Yang Miscellaneous Notes)].

    PubMed

    Zhang, Jun-Ke

    2010-09-01

    Guangyangzaji (Guang Yang Miscellaneous Notes) are historical notes written by a scholar of the Qing dynasty: Liu Xian-ting. This book recorded much medical content of the people. Doing simple research and analysis on the name, physical nature and effectiveness of the Lobular Pyrolae, Kai Wood (Chinese pistache), Huanglian snakes (a snake habitat with Coptis grass) and other native herbs can contribute to understanding the evolution of the medicinal herbs. PMID:21163084

  7. Robust and tunable circadian rhythms from differentially sensitive catalytic domains

    PubMed Central

    Phong, Connie; Markson, Joseph S.; Wilhoite, Crystal M.; Rust, Michael J.

    2013-01-01

    Circadian clocks are ubiquitous biological oscillators that coordinate an organism’s behavior with the daily cycling of the external environment. To ensure synchronization with the environment, the period of the clock must be maintained near 24 h even as amplitude and phase are altered by input signaling. We show that, in a reconstituted circadian system from cyanobacteria, these conflicting requirements are satisfied by distinct functions for two domains of the central clock protein KaiC: the C-terminal autokinase domain integrates input signals through the ATP/ADP ratio, and the slow N-terminal ATPase acts as an input-independent timer. We find that phosphorylation in the C-terminal domain followed by an ATPase cycle in the N-terminal domain is required to form the inhibitory KaiB•KaiC complexes that drive the dynamics of the clock. We present a mathematical model in which this ATPase-mediated delay in negative feedback gives rise to a compensatory mechanism that allows a tunable phase and amplitude while ensuring a robust circadian period. PMID:23277568

  8. Robust and tunable circadian rhythms from differentially sensitive catalytic domains.

    PubMed

    Phong, Connie; Markson, Joseph S; Wilhoite, Crystal M; Rust, Michael J

    2013-01-15

    Circadian clocks are ubiquitous biological oscillators that coordinate an organism's behavior with the daily cycling of the external environment. To ensure synchronization with the environment, the period of the clock must be maintained near 24 h even as amplitude and phase are altered by input signaling. We show that, in a reconstituted circadian system from cyanobacteria, these conflicting requirements are satisfied by distinct functions for two domains of the central clock protein KaiC: the C-terminal autokinase domain integrates input signals through the ATP/ADP ratio, and the slow N-terminal ATPase acts as an input-independent timer. We find that phosphorylation in the C-terminal domain followed by an ATPase cycle in the N-terminal domain is required to form the inhibitory KaiB•KaiC complexes that drive the dynamics of the clock. We present a mathematical model in which this ATPase-mediated delay in negative feedback gives rise to a compensatory mechanism that allows a tunable phase and amplitude while ensuring a robust circadian period. PMID:23277568

  9. Theophylline-dependent riboswitch as a novel genetic tool for strict regulation of protein expression in Cyanobacterium Synechococcus elongatus PCC 7942.

    PubMed

    Nakahira, Yoichi; Ogawa, Atsushi; Asano, Hiroyuki; Oyama, Tokitaka; Tozawa, Yuzuru

    2013-10-01

    The cyanobacterium Synechococcus elongatus PCC 7942 is a major model species for studies of photosynthesis. It is are also a potential cell factory for the production of renewable biofuels and valuable chemicals. We employed engineered riboswitches to control translational initiation of target genes in this cyanobacterium. A firefly luciferase reporter assay revealed that three theophylline riboswitches performed as expected in the cyanobacterium. Riboswitch-E* exhibited very low leaky expression of luciferase and superior and dose-dependent on/off regulation of protein expression by theophylline. The maximum magnitude of the induction vs. basal level was ∼190-fold. Furthermore, the induction level was responsive to a wide range of theophylline concentrations in the medium, from 0 to 2 mM, facilitating the fine-tuning of luciferase expression. We adapted this riboswitch to another gene regulation system, in which expression of the circadian clock kaiC gene product is controlled by the theophylline concentration in the culture medium. The results demonstrated that the adequately adjusted expression level of KaiC restored complete circadian rhythm in the kaiC-deficient arrhythmic mutant. This theophylline-dependent riboswitch system has potential for various applications as a useful genetic tool in cyanobacteria. PMID:23969558

  10. Daily Rhythms in the Cyanobacterium Synechococcus elongatus Probed by High-resolution Mass Spectrometry–based Proteomics Reveals a Small Defined Set of Cyclic Proteins*

    PubMed Central

    Guerreiro, Ana C. L.; Benevento, Marco; Lehmann, Robert; van Breukelen, Bas; Post, Harm; Giansanti, Piero; Maarten Altelaar, A. F.; Axmann, Ilka M.; Heck, Albert J. R.

    2014-01-01

    Circadian rhythms are self-sustained and adjustable cycles, typically entrained with light/dark and/or temperature cycles. These rhythms are present in animals, plants, fungi, and several bacteria. The central mechanism behind these “pacemakers” and the connection to the circadian regulated pathways are still poorly understood. The circadian rhythm of the cyanobacterium Synechococcus elongatus PCC 7942 (S. elongatus) is highly robust and controlled by only three proteins, KaiA, KaiB, and KaiC. This central clock system has been extensively studied functionally and structurally and can be reconstituted in vitro. These characteristics, together with a relatively small genome (2.7 Mbp), make S. elongatus an ideal model system for the study of circadian rhythms. Different approaches have been used to reveal the influence of the central S. elongatus clock on rhythmic gene expression, rhythmic mRNA abundance, rhythmic DNA topology changes, and cell division. However, a global analysis of its proteome dynamics has not been reported yet. To uncover the variation in protein abundances during 48 h under light and dark cycles (12:12 h), we used quantitative proteomics, with TMT 6-plex isobaric labeling. We queried the S. elongatus proteome at 10 different time points spanning a single 24-h period, leading to 20 time points over the full 48-h period. Employing multidimensional separation and high-resolution mass spectrometry, we were able to find evidence for a total of 82% of the S. elongatus proteome. Of the 1537 proteins quantified over the time course of the experiment, only 77 underwent significant cyclic variations. Interestingly, our data provide evidence for in- and out-of-phase correlation between mRNA and protein levels for a set of specific genes and proteins. As a range of cyclic proteins are functionally not well annotated, this work provides a resource for further studies to explore the role of these proteins in the cyanobacterial circadian rhythm. PMID

  11. Using Information and Communications Technology in a National Population-Based Survey: The Kenya AIDS Indicator Survey 2012

    PubMed Central

    Ojwang’, James K.; Lee, Veronica C.; Waruru, Anthony; Ssempijja, Victor; Ng’ang’a, John G.; Wakhutu, Brian E.; Kandege, Nicholas O.; Koske, Danson K.; Kamiru, Samuel M.; Omondi, Kenneth O.; Kakinyi, Mutua; Kim, Andrea A.; Oluoch, Tom

    2016-01-01

    Background With improvements in technology, electronic data capture (EDC) for large surveys is feasible. EDC offers benefits over traditional paper-based data collection, including more accurate data, greater completeness of data, and decreased data cleaning burden. Methods The second Kenya AIDS Indicator Survey (KAIS 2012) was a population-based survey of persons aged 18 months to 64 years. A software application was designed to capture the interview, specimen collection, and home-based testing and counseling data. The application included: interview translations for local languages; options for single, multiple, and fill-in responses; and automated participant eligibility determination. Data quality checks were programmed to automate skip patterns and prohibit outlier responses. A data sharing architecture was developed to transmit the data in realtime from the field to a central server over a virtual private network. Results KAIS 2012 was conducted between October 2012 and February 2013. Overall, 68,202 records for the interviews, specimen collection, and home-based testing and counseling were entered into the application. Challenges arose during implementation, including poor connectivity and a systems malfunction that created duplicate records, which prevented timely data transmission to the central server. Data cleaning was minimal given the data quality control measures. Conclusions KAIS 2012 demonstrated the feasibility of using EDC in a population-based survey. The benefits of EDC were apparent in data quality and minimal time needed for data cleaning. Several important lessons were learned, such as the time and monetary investment required before survey implementation, the importance of continuous application testing, and contingency plans for data transmission due to connectivity challenges. PMID:24732816

  12. Seroprevalence of Infections with Dengue, Rift Valley Fever and Chikungunya Viruses in Kenya, 2007

    PubMed Central

    Ochieng, Caroline; Ahenda, Petronella; Vittor, Amy Y.; Nyoka, Raymond; Gikunju, Stella; Wachira, Cyrus; Waiboci, Lilian; Umuro, Mamo; Kim, Andrea A.; Nderitu, Leonard; Juma, Bonventure; Montgomery, Joel M.; Breiman, Robert F.; Fields, Barry

    2015-01-01

    Arthropod-borne viruses are a major constituent of emerging infectious diseases worldwide, but limited data are available on the prevalence, distribution, and risk factors for transmission in Kenya and East Africa. In this study, we used 1,091 HIV-negative blood specimens from the 2007 Kenya AIDS Indicator Survey (KAIS 2007) to test for the presence of IgG antibodies to dengue virus (DENV), chikungunya virus (CHIKV) and Rift Valley fever virus (RVFV).The KAIS 2007 was a national population-based survey conducted by the Government of Kenya to provide comprehensive information needed to address the HIV/AIDS epidemic. Antibody testing for arboviruses was performed on stored blood specimens from KAIS 2007 through a two-step sandwich IgG ELISA using either commercially available kits or CDC-developed assays. Out of the 1,091 samples tested, 210 (19.2%) were positive for IgG antibodies against at least one of the three arboviruses. DENV was the most common of the three viruses tested (12.5% positive), followed by RVFV and CHIKV (4.5% and 0.97%, respectively). For DENV and RVFV, the participant’s province of residence was significantly associated (P≤.01) with seropositivity. Seroprevalence of DENV and RVFV increased with age, while there was no correlation between province of residence/age and seropositivity for CHIKV. Females had twelve times higher odds of exposure to CHIK as opposed to DENV and RVFV where both males and females had the same odds of exposure. Lack of education was significantly associated with a higher odds of previous infection with either DENV or RVFV (p <0.01). These data show that a number of people are at risk of arbovirus infections depending on their geographic location in Kenya and transmission of these pathogens is greater than previously appreciated. This poses a public health risk, especially for DENV. PMID:26177451

  13. Diurnal rhythm of a unicellular diazotrophic cyanobacterium under mixotrophic conditions and elevated carbon dioxide.

    PubMed

    Gaudana, Sandeep B; Alagesan, Swathi; Chetty, Madhu; Wangikar, Pramod P

    2013-11-01

    Mixotrophic cultivation of cyanobacteria in wastewaters with flue gas sparging has the potential to simultaneously sequester carbon content from gaseous and aqueous streams and convert to biomass and biofuels. Therefore, it was of interest to study the effect of mixotrophy and elevated CO2 on metabolism, morphology and rhythm of gene expression under diurnal cycles. We chose a diazotrophic unicellular cyanobacterium Cyanothece sp. ATCC 51142 as a model, which is a known hydrogen producer with robust circadian rhythm. Cyanothece 51142 grows faster with nitrate and/or an additional carbon source in the growth medium and at 3 % CO2. Intracellular glycogen contents undergo diurnal oscillations with greater accumulation under mixotrophy. While glycogen is exhausted by midnight under autotrophic conditions, significant amounts remain unutilized accompanied by a prolonged upregulation of nifH gene under mixotrophy. This possibly supports nitrogen fixation for longer periods thereby leading to better growth. To gain insights into the influence of mixotrophy and elevated CO2 on circadian rhythm, transcription of core clock genes kaiA, kaiB1 and kaiC1, the input pathway, cikA, output pathway, rpaA and representatives of key metabolic pathways was analyzed. Clock genes' transcripts were lower under mixotrophy suggesting a dampening effect exerted by an external carbon source such as glycerol. Nevertheless, the genes of the clock and important metabolic pathways show diurnal oscillations in expression under mixotrophic and autotrophic growth at ambient and elevated CO2, respectively. Taken together, the results indicate segregation of light and dark associated reactions even under mixotrophy and provide important insights for further applications. PMID:23881383

  14. Estimate of transport properties of porous media by microfocus X-ray computed tomography and random walk simulation

    NASA Astrophysics Data System (ADS)

    Nakashima, Yoshito; Watanabe, Yoshinori

    2002-12-01

    The transport properties (porosity, surface-to-volume ratio of the pore space, diffusion coefficient, and permeability) of a porous medium were calculated by image analysis and random walk simulation using the digital image data on the pore structure of a bead pack (diameter 2.11 mm). A theory developed for laboratory experiments of nuclear magnetic resonance was applied to the random walk simulation. The three-dimensional data set (2563 voxels) of the bead pack was obtained by microfocus X-ray computed tomography at a spatial resolution of 0.053 mm. An original cluster labeling program, Kai3D.m, was used to estimate the porosity and surface-to-volume ratio. The surface-to-volume ratio and diffusion coefficient were calculated by an original random walk program, RW3D.m. The calculations were completed on a personal computer in reasonable time (≤13 hours). The permeability was estimated by substituting the results of Kai3D.m and RW3D.m into the Kozeny-Carman equation. The results for the porosity, surface-to-volume ratio, and diffusion coefficient were within 5-8% of measured values, whereas the calculated permeability involved an error of 35%. The promising results of the present study indicate that it is possible to estimate the permeability of porous media with reasonable accuracy by the diffusometry and random walk simulation. Because, in principle, the diffusometry could be performed by proton nuclear magnetic resonance logging, the method of estimating the transport properties presented here is applicable to the in situ measurement of strata. We open the original Mathematica® programs (Kai3D.m and RW3D.m) used to calculate the porosity, surface-to-volume ratio, and diffusion coefficient at the authors' home page to facilitate the personal-computer-based study of porous media using X-ray computed tomography.

  15. Design of multi-mode compatible image acquisition system for HD area array CCD

    NASA Astrophysics Data System (ADS)

    Wang, Chen; Sui, Xiubao

    2014-11-01

    Combining with the current development trend in video surveillance-digitization and high-definition, a multimode-compatible image acquisition system for HD area array CCD is designed. The hardware and software designs of the color video capture system of HD area array CCD KAI-02150 presented by Truesense Imaging company are analyzed, and the structure parameters of the HD area array CCD and the color video gathering principle of the acquisition system are introduced. Then, the CCD control sequence and the timing logic of the whole capture system are realized. The noises of the video signal (KTC noise and 1/f noise) are filtered by using the Correlated Double Sampling (CDS) technique to enhance the signal-to-noise ratio of the system. The compatible designs in both software and hardware for the two other image sensors of the same series: KAI-04050 and KAI-08050 are put forward; the effective pixels of these two HD image sensors are respectively as many as four million and eight million. A Field Programmable Gate Array (FPGA) is adopted as the key controller of the system to perform the modularization design from top to bottom, which realizes the hardware design by software and improves development efficiency. At last, the required time sequence driving is simulated accurately by the use of development platform of Quartus II 12.1 combining with VHDL. The result of the simulation indicates that the driving circuit is characterized by simple framework, low power consumption, and strong anti-interference ability, which meet the demand of miniaturization and high-definition for the current tendency.

  16. A design of driving circuit for star sensor imaging camera

    NASA Astrophysics Data System (ADS)

    Li, Da-wei; Yang, Xiao-xu; Han, Jun-feng; Liu, Zhao-hui

    2016-01-01

    The star sensor is a high-precision attitude sensitive measuring instruments, which determine spacecraft attitude by detecting different positions on the celestial sphere. Imaging camera is an important portion of star sensor. The purpose of this study is to design a driving circuit based on Kodak CCD sensor. The design of driving circuit based on Kodak KAI-04022 is discussed, and the timing of this CCD sensor is analyzed. By the driving circuit testing laboratory and imaging experiments, it is found that the driving circuits can meet the requirements of Kodak CCD sensor.

  17. In-situ demonstration of radio-frequency enhanced chlorinated hydrocarbon remediation

    SciTech Connect

    Kasevich, R.S.; Price, S.L.; Faust, D.L.; Jarosch, T.R.

    1994-06-01

    This paper discusses the results of a successful demonstration of radio frequency (RF) heating for enhanced chlorinated hydrocarbon remediation at the M-Area Seepage Basin of the Department of Energy`s Savannah River Site. RF heating was integrated with soil vapor extraction (SVE) to enhance the release of residual volatile chlorinated hydrocarbons which are concentrated in low permeable clay lenses in the unsaturated zone. Participants in this effort consisted of the Westinghouse Savannah River Technology Center; the Westinghouse Science and Technology Center (Pittsburgh, PA); and KAI Technologies, Inc. which provided the RF technology. Additionally, a better understanding of RF heating technology is gained through a description of the RF heating system.

  18. [The Evolutionary Origin of Placodes and Neural Crest Cells

    NASA Technical Reports Server (NTRS)

    Bronner-Fraser, Marianne

    2003-01-01

    The long-term goal of this NASA-supported research is to understand the evolutionary origin of placodes and neural crest cells, with particular reference to evolution of the inner ear, and their evolutionary and developmental relationships. The cephalochordcate amphioxus, the closest living invertebrate relative of the vertebrates is used as a stand-in for the ancestral vertebrate. The research, which has supported one graduate student, Jr-Kai Yu, has resulted in ten publications by the Holland laboratory in peer-reviewed journals.

  19. Present status of information centers (libraries)

    NASA Astrophysics Data System (ADS)

    Suzuki, Keigo; Adachi, Takashi; Kusumi, Yoshihisa; Nishiyama, Mieko; Hirata, Toshiichiro

    Documentation Kondan-Kai (gathering for discussing issues) has conducted the questionnaire which asks present status of information centers of companies. The purpose of this questionnaire was to obtain some tips which would be useful for companies to consider what their information centers should be in a highly informationalized society, and how they should be changing. The results were grouped into the following five sections and analyzed; (1) management system of information center, (2) use condition of information center, (3) management of books and journals, (4) external online information retrieval and (5) management of technical materials generated inside company.

  20. Detectable HIV Viral Load in Kenya: Data from a Population-Based Survey

    PubMed Central

    Cherutich, Peter; Kim, Andrea A.; Kellogg, Timothy A.; Sherr, Kenneth; Waruru, Anthony; De Cock, Kevin M.; Rutherford, George W.

    2016-01-01

    Introduction At the individual level, there is clear evidence that Human Immunodeficiency Virus (HIV) transmission can be substantially reduced by lowering viral load. However there are few data describing population-level HIV viremia especially in high-burden settings with substantial under-diagnosis of HIV infection. The 2nd Kenya AIDS Indicator Survey (KAIS 2012) provided a unique opportunity to evaluate the impact of antiretroviral therapy (ART) coverage on viremia and to examine the risks for failure to suppress viral replication. We report population-level HIV viral load suppression using data from KAIS 2012. Methods Between October 2012 to February 2013, KAIS 2012 surveyed household members, administered questionnaires and drew serum samples to test for HIV and, for those found to be infected with HIV, plasma viral load (PVL) was measured. Our principal outcome was unsuppressed HIV viremia, defined as a PVL ≥ 550 copies/mL. The exposure variables included current treatment with ART, prior history of an HIV diagnosis, and engagement in HIV care. All point estimates were adjusted to account for the KAIS 2012 cluster sampling design and survey non-response. Results Overall, 61·2% (95% CI: 56·4–66·1) of HIV-infected Kenyans aged 15–64 years had not achieved virological suppression. The base10 median (interquartile range [IQR]) and mean (95% CI) VL was 4,633 copies/mL (0–51,596) and 81,750 copies/mL (59,366–104,134), respectively. Among 266 persons taking ART, 26.1% (95% CI: 20.0–32.1) had detectable viremia. Non-ART use, younger age, and lack of awareness of HIV status were independently associated with significantly higher odds of detectable viral load. In multivariate analysis for the sub-sample of patients on ART, detectable viremia was independently associated with younger age and sub-optimal adherence to ART. Discussion This report adds to the limited data of nationally-representative surveys to report population- level virological

  1. [A brief history of Chinese royal court medicines].

    PubMed

    Li, Chun-Sheng

    2010-09-01

    With the establishment of an emperor and a royal court, the court physician came into being and the royal court medicines gradually began to evolve. In the first year of Kai Huang of the Sui dynasty (581), King Wendi of the Sui dynasty established the imperial medical bureau. Since then the royal court medicines entered a period of development and prosperity. In the Yuan dynasty, the scope of official duty of the imperial hospital narrowed, the development of royal court medicines lacked new growth. To the Ming and Qing dynasties, the royal court medicine began to decline and eventually ended with the demise of the Qing dynasty. PMID:21163077

  2. Genome-Wide and Heterocyst-Specific Circadian Gene Expression in the Filamentous Cyanobacterium Anabaena sp. Strain PCC 7120

    PubMed Central

    Kushige, Hiroko; Kugenuma, Hideyuki; Matsuoka, Masaki; Ehira, Shigeki; Ohmori, Masayuki

    2013-01-01

    The filamentous, heterocystous cyanobacterium Anabaena sp. strain PCC 7120 is one of the simplest multicellular organisms that show both morphological pattern formation with cell differentiation (heterocyst formation) and circadian rhythms. Therefore, it potentially provides an excellent model in which to analyze the relationship between circadian functions and multicellularity. However, detailed cyanobacterial circadian regulation has been intensively analyzed only in the unicellular species Synechococcus elongatus. In contrast to the highest-amplitude cycle in Synechococcus, we found that none of the kai genes in Anabaena showed high-amplitude expression rhythms. Nevertheless, ∼80 clock-controlled genes were identified. We constructed luciferase reporter strains to monitor the expression of some high-amplitude genes. The bioluminescence rhythms satisfied the three criteria for circadian oscillations and were nullified by genetic disruption of the kai gene cluster. In heterocysts, in which photosystem II is turned off, the metabolic and redox states are different from those in vegetative cells, although these conditions are thought to be important for circadian entrainment and timekeeping processes. Here, we demonstrate that circadian regulation is active in heterocysts, as shown by the finding that heterocyst-specific genes, such as all1427 and hesAB, are expressed in a robust circadian fashion exclusively without combined nitrogen. PMID:23316037

  3. Crystal structure of the flagellar accessory protein FlaH of Methanocaldococcus jannaschii suggests a regulatory role in archaeal flagellum assembly.

    PubMed

    Meshcheryakov, Vladimir A; Wolf, Matthias

    2016-06-01

    Archaeal flagella are unique structures that share functional similarity with bacterial flagella, but are structurally related to bacterial type IV pili. The flagellar accessory protein FlaH is one of the conserved components of the archaeal motility system. However, its function is not clearly understood. Here, we present the 2.2 Å resolution crystal structure of FlaH from the hyperthermophilic archaeon, Methanocaldococcus jannaschii. The protein has a characteristic RecA-like fold, which has been found previously both in archaea and bacteria. We show that FlaH binds to immobilized ATP-however, it lacks ATPase activity. Surface plasmon resonance analysis demonstrates that ATP affects the interaction between FlaH and the archaeal motor protein FlaI. In the presence of ATP, the FlaH-FlaI interaction becomes significantly weaker. A database search revealed similarity between FlaH and several DNA-binding proteins of the RecA superfamily. The closest structural homologs of FlaH are KaiC-like proteins, which are archaeal homologs of the circadian clock protein KaiC from cyanobacteria. We propose that one of the functions of FlaH may be the regulation of archaeal motor complex assembly. PMID:27060465

  4. Aberration analysis and calculation in system of Gaussian beam illuminates lenslet array

    NASA Astrophysics Data System (ADS)

    Zhao, Zhu; Hui, Mei; Zhou, Ping; Su, Tianquan; Feng, Yun; Zhao, Yuejin

    2014-09-01

    Low order aberration was founded when focused Gaussian beam imaging at Kodak KAI -16000 image detector, which is integrated with lenslet array. Effect of focused Gaussian beam and numerical simulation calculation of the aberration were presented in this paper. First, we set up a model of optical imaging system based on previous experiment. Focused Gaussian beam passed through a pinhole and was received by Kodak KAI -16000 image detector whose microlenses of lenslet array were exactly focused on sensor surface. Then, we illustrated the characteristics of focused Gaussian beam and the effect of relative space position relations between waist of Gaussian beam and front spherical surface of microlenses to the aberration. Finally, we analyzed the main element of low order aberration and calculated the spherical aberration caused by lenslet array according to the results of above two steps. Our theoretical calculations shown that , the numerical simulation had a good agreement with the experimental result. Our research results proved that spherical aberration was the main element and made up about 93.44% of the 48 nm error, which was demonstrated in previous experiment. The spherical aberration is inversely proportional to the value of divergence distance between microlens and waist, and directly proportional to the value of the Gaussian beam waist radius.

  5. Driving techniques for high frame rate CCD camera

    NASA Astrophysics Data System (ADS)

    Guo, Weiqiang; Jin, Longxu; Xiong, Jingwu

    2008-03-01

    This paper describes a high-frame rate CCD camera capable of operating at 100 frames/s. This camera utilizes Kodak KAI-0340, an interline transfer CCD with 640(vertical)×480(horizontal) pixels. Two output ports are used to read out CCD data and pixel rates approaching 30 MHz. Because of its reduced effective opacity of vertical charge transfer registers, interline transfer CCD can cause undesired image artifacts, such as random white spots and smear generated in the registers. To increase frame rate, a kind of speed-up structure has been incorporated inside KAI-0340, then it is vulnerable to a vertical stripe effect. The phenomena which mentioned above may severely impair the image quality. To solve these problems, some electronic methods of eliminating these artifacts are adopted. Special clocking mode can dump the unwanted charge quickly, then the fast readout of the images, cleared of smear, follows immediately. Amplifier is used to sense and correct delay mismatch between the dual phase vertical clock pulses, the transition edges become close to coincident, so vertical stripes disappear. Results obtained with the CCD camera are shown.

  6. Lichen Secondary Metabolite, Physciosporin, Inhibits Lung Cancer Cell Motility.

    PubMed

    Yang, Yi; Park, So-Yeon; Nguyen, Thanh Thi; Yu, Young Hyun; Nguyen, Tru Van; Sun, Eun Gene; Udeni, Jayalal; Jeong, Min-Hye; Pereira, Iris; Moon, Cheol; Ha, Hyung-Ho; Kim, Kyung Keun; Hur, Jae-Seoun; Kim, Hangun

    2015-01-01

    Lichens produce various unique chemicals that can be used for pharmaceutical purposes. To screen for novel lichen secondary metabolites showing inhibitory activity against lung cancer cell motility, we tested acetone extracts of 13 lichen samples collected in Chile. Physciosporin, isolated from Pseudocyphellaria coriacea (Hook f. & Taylor) D.J. Galloway & P. James, was identified as an effective compound and showed significant inhibitory activity in migration and invasion assays against human lung cancer cells. Physciosporin treatment reduced both protein and mRNA levels of N-cadherin with concomitant decreases in the levels of epithelial-mesenchymal transition markers such as snail and twist. Physciosporin also suppressed KITENIN (KAI1 C-terminal interacting tetraspanin)-mediated AP-1 activity in both the absence and presence of epidermal growth factor stimulation. Quantitative real-time PCR analysis showed that the expression of the metastasis suppressor gene, KAI1, was increased while that of the metastasis enhancer gene, KITENIN, was dramatically decreased by physciosporin. Particularly, the activity of 3'-untranslated region of KITENIN was decreased by physciosporin. Moreover, Cdc42 and Rac1 activities were decreased by physciosporin. These results demonstrated that the lichen secondary metabolite, physciosporin, inhibits lung cancer cell motility through novel mechanisms of action. PMID:26371759

  7. Lichen Secondary Metabolite, Physciosporin, Inhibits Lung Cancer Cell Motility

    PubMed Central

    Yang, Yi; Park, So-Yeon; Nguyen, Thanh Thi; Yu, Young Hyun; Nguyen, Tru Van; Sun, Eun Gene; Udeni, Jayalal; Jeong, Min-Hye; Pereira, Iris; Moon, Cheol; Ha, Hyung-Ho; Kim, Kyung Keun; Hur, Jae-Seoun; Kim, Hangun

    2015-01-01

    Lichens produce various unique chemicals that can be used for pharmaceutical purposes. To screen for novel lichen secondary metabolites showing inhibitory activity against lung cancer cell motility, we tested acetone extracts of 13 lichen samples collected in Chile. Physciosporin, isolated from Pseudocyphellaria coriacea (Hook f. & Taylor) D.J. Galloway & P. James, was identified as an effective compound and showed significant inhibitory activity in migration and invasion assays against human lung cancer cells. Physciosporin treatment reduced both protein and mRNA levels of N-cadherin with concomitant decreases in the levels of epithelial-mesenchymal transition markers such as snail and twist. Physciosporin also suppressed KITENIN (KAI1 C-terminal interacting tetraspanin)-mediated AP-1 activity in both the absence and presence of epidermal growth factor stimulation. Quantitative real-time PCR analysis showed that the expression of the metastasis suppressor gene, KAI1, was increased while that of the metastasis enhancer gene, KITENIN, was dramatically decreased by physciosporin. Particularly, the activity of 3’-untranslated region of KITENIN was decreased by physciosporin. Moreover, Cdc42 and Rac1 activities were decreased by physciosporin. These results demonstrated that the lichen secondary metabolite, physciosporin, inhibits lung cancer cell motility through novel mechanisms of action. PMID:26371759

  8. Art, class and gender in Joseon dynasty Korea: representations of lower-class women by the scholar-painter Yun Duseo.

    PubMed

    Chung, Saehyang P

    2011-01-01

    This paper examines several pioneering genre paintings by the important scholar painter Yun Duseo (1668-1715), with its focus on their artistic sources which have not yet been explored so far. Painted on ramie, 'Women Picking Potherbs' is one of the most intriguing examples among Yun Duseo's oeuvre, which encompasses a broad variety of themes, including genre imagery, landscapes, portraits, dragons, and horses. Even among Yun Duseo's genre paintings, 'Women Picking Potherbs' is extraordinary, as recent scholarship regards it as the earliest independent representation of lower-class women in the history of Korean art. In particular, Yun Duseo painted two women who were working ourdoors to gather spring potherbs. In a conservative Confucian society, it was extraordinary women who were working outdoors. Hence, Yun Duseo occupies a highly important place in Korean painting. Furthermore, even though Yun Duseo came from the upper-class, he often painted images of lower class people working. It is possible that Yun Duseo was familiar with the book titled "Tian gong kai wu" (Exploitation of the Works of Nature) which was published in the 17th century. By identifying the probable body of his artistic sources in the book known as "Tian gong kai wu," it will be possible to assess the innovations and limitations found in 'Women Picking Potherbs'. PMID:22171413

  9. Exploration of the factor structure of the Kirton Adaption-Innovation Inventory using bootstrapping estimation.

    PubMed

    Im, Subin; Min, Soonhong

    2013-04-01

    Exploratory factor analyses of the Kirton Adaption-Innovation Inventory (KAI), which serves to measure individual cognitive styles, generally indicate three factors: sufficiency of originality, efficiency, and rule/group conformity. In contrast, a 2005 study by Im and Hu using confirmatory factor analysis supported a four-factor structure, dividing the sufficiency of originality dimension into two subdimensions, idea generation and preference for change. This study extends Im and Hu's (2005) study of a derived version of the KAI by providing additional evidence of the four-factor structure. Specifically, the authors test the robustness of the parameter estimates to the violation of normality assumptions in the sample using bootstrap methods. A bias-corrected confidence interval bootstrapping procedure conducted among a sample of 356 participants--members of the Arkansas Household Research Panel, with middle SES and average age of 55.6 yr. (SD = 13.9)--showed that the four-factor model with two subdimensions of sufficiency of originality fits the data significantly better than the three-factor model in non-normality conditions. PMID:23833873

  10. Compact Sources of Ultrashort Pulses

    NASA Astrophysics Data System (ADS)

    Duling, Irl N., III

    2006-11-01

    List of contributors; Acronyms and abbreviations; Preface; 1. Short pulse generation H. A. Haus; 2. Passive modelocking in solid state lasers Thomas Brabec, Stephen M. J. Kelly and Ferenc Krausz; 3. Compact modelocked solid state lasers pumped by laser diodes John R. M. Barr; 4. Modelocking of all-fiber lasers Irl N. Duling, III and Michael L. Dennis; 5. Nonlinear polarization evolution in passively modelocked fiber lasers Martin E. Fermann; 6. Ultrafast vertical cavity semiconductor lasers Wenbin Jiang and John Bowers; 7. High power ultrafast semiconductor for injection diode lasers Peter J. Delfyett; 8. The hybrid soliton pulse source Paul A. Morton; 9. Monolithic colliding pulse modelocked diode lasers Ming C. Wu and Young-Kai Chen; Index.

  11. Hierarchy of models: From qualitative to quantitative analysis of circadian rhythms in cyanobacteria

    NASA Astrophysics Data System (ADS)

    Chaves, M.; Preto, M.

    2013-06-01

    A hierarchy of models, ranging from high to lower levels of abstraction, is proposed to construct "minimal" but predictive and explanatory models of biological systems. Three hierarchical levels will be considered: Boolean networks, piecewise affine differential (PWA) equations, and a class of continuous, ordinary, differential equations' models derived from the PWA model. This hierarchy provides different levels of approximation of the biological system and, crucially, allows the use of theoretical tools to more exactly analyze and understand the mechanisms of the system. The Kai ABC oscillator, which is at the core of the cyanobacterial circadian rhythm, is analyzed as a case study, showing how several fundamental properties—order of oscillations, synchronization when mixing oscillating samples, structural robustness, and entrainment by external cues—can be obtained from basic mechanisms.

  12. History of Amateur Variable Star Observations in Japan

    NASA Astrophysics Data System (ADS)

    Kiyota, S.

    2012-06-01

    (Abstract only) Japan has about 100 years of history of variable star observing since Naozo Ichinohe, professional astronomer in Tokyo Observatory, observed d Cep in 1906. The first amateur variable star observer is Yoshihiko Kasai, who began observing variable stars in 1918. I introduce a brief history of Japanese amateur variable star observation, including topics of variable star organizations, nova and supernova hunters, collaborations with the AAVSO and the world, PEP and CCD observations. I also introduce the most active variable star observer, Hiroaki Narumi, who made over 260,000 visual estimates since 1975. VSOLJ was established in 1987 in collaborations with the variable star sections of Nihon Tenmon Kenkyu-kai (NTK) and the Oriental Astronomical Association (OAA). VSOLJ maintains a database of Japanese variable star observations (http://vsolj.cetus-net.org) and publishes the Variable Star Bulletin in English.

  13. Filtration method efficiently desalts crude in commercial test

    SciTech Connect

    Not Available

    1993-05-17

    During 3 months of industrial testing of a filtration crude oil desalting method, a total of 120,500 metric tons (mt), or 1,475 mt/d (almost 11,000 b/d) of crude was processed. Rongxi Du, Kai Peng, and Li Wang, engineers at Wuhan Petrochemical Works, Wuhan, China, in an unpublished report, indicate that they determined unit operating parameters and performed statistical analyses of desalting-efficiency data from the test run. The engineers also determined relationships between desalting efficiency and flow velocity, relative density, mixing pressure drop (MPD), filtration-tank pressure drop, and temperature. The desalting and dewatering level of single-stage filtrations desalting was found to be equal to that of two-stage electrostatic desalting with remarkable benefits resulting from reduced power, water, and demulsifier requirements. This paper describes the filtration desalting, test parameters, performance results, and filter revivification.

  14. Technical evaluation report for the demonstration of radio frequency soil decontamination at Site S-1

    SciTech Connect

    Lyon, C.R.; Blanchard, C.F.; Whitt, L.H.

    1995-04-01

    The Air Force`s Armstrong Laboratory at Tyndall Air Force Base, Florida, has supported the research and development of Radio Frequency Soil Decontamination. Radio frequency soil decontamination is essentially a heat-assisted soil vapor extraction process. Site S-1 at Kelly Air Force Base, San Antonio, Texas, was selected for the demonstration of two patented techniques. The site is a former sump that collected spills and surface run-off from a waste petroleum, oils, and lubricants and solvent storage and transfer area. In 1993, a technique developed by the IIT Research Institute using an array of electrodes placed in the soil was demonstrated. In 1994, a technique developed by KAI Technologies, Inc. using a single applicator placed in a vertical borehole was demonstrated. Approximately 120 tons of soil were heated during each demonstration to a temperature of about 150 degrees Celsius.

  15. Using adjuvants and environmental factors to modulate the activity of antimicrobial peptides.

    PubMed

    Walkenhorst, William F

    2016-05-01

    The increase in antibiotic resistant and multi-drug resistant bacterial infections has serious implications for the future of health care. The difficulty in finding both new microbial targets and new drugs against existing targets adds to the concern. The use of combination and adjuvant therapies are potential strategies to counter this threat. Antimicrobial peptides (AMPs) are a promising class of antibiotics (ABs), particularly for topical and surface applications. Efforts have been directed toward a number of strategies, including the use of conventional ABs combined with AMPs, and the use of potentiating agents to increase the performance of AMPs. This review focuses on combination strategies such as adjuvants and the manipulation of environmental variables to improve the efficacy of AMPs as potential therapeutic agents. This article is part of a Special Issue entitled: Antimicrobial peptides edited by Karl Lohner and Kai Hilpert. PMID:26751595

  16. Helicopter fuselage drag - combined computational fluid dynamics and experimental studies

    NASA Astrophysics Data System (ADS)

    Batrakov, A.; Kusyumov, A.; Mikhailov, S.; Pakhov, V.; Sungatullin, A.; Valeev, M.; Zherekhov, V.; Barakos, G.

    2015-06-01

    In this paper, wind tunnel experiments are combined with Computational Fluid Dynamics (CFD) aiming to analyze the aerodynamics of realistic fuselage configurations. A development model of the ANSAT aircraft and an early model of the AKTAI light helicopter were employed. Both models were tested at the subsonic wind tunnel of KNRTU-KAI for a range of Reynolds numbers and pitch and yaw angles. The force balance measurements were complemented by particle image velocimetry (PIV) investigations for the cases where the experimental force measurements showed substantial unsteadiness. The CFD results were found to be in fair agreement with the test data and revealed some flow separation at the rear of the fuselages. Once confidence on the CFD method was established, further modifications were introduced to the ANSAT-like fuselage model to demonstrate drag reduction via small shape changes.

  17. Application of Barcoding to Reduce Error of Patient Identification and to Increase Patient's Information Confidentiality of Test Tube Labelling in a Psychiatric Teaching Hospital.

    PubMed

    Liu, Hsiu-Chu; Li, Hsing; Chang, Hsin-Fei; Lu, Mei-Rou; Chen, Feng-Chuan

    2015-01-01

    Learning from the experience of another medical center in Taiwan, Kaohsiung Municipal Kai-Suan Psychiatric Hospital has changed the nursing informatics system step by step in the past year and a half . We considered ethics in the original idea of implementing barcodes on the test tube labels to process the identification of the psychiatric patients. The main aims of this project are to maintain the confidential information and to transport the sample effectively. The primary nurses had been using different work sheets for this project to ensure the acceptance of the new barcode system. In the past two years the errors in the blood testing process were as high as 11,000 in 14,000 events per year, resulting in wastage of resources. The actions taken by the nurses and the new barcode system implementation can improve the clinical nursing care quality, safety of the patients, and efficiency, while decreasing the cost due to the human error. PMID:26262221

  18. Synthetic antibiofilm peptides.

    PubMed

    de la Fuente-Núñez, César; Cardoso, Marlon Henrique; de Souza Cândido, Elizabete; Franco, Octavio Luiz; Hancock, Robert E W

    2016-05-01

    Bacteria predominantly exist as multicellular aggregates known as biofilms that are associated with at least two thirds of all infections and exhibit increased adaptive resistance to conventional antibiotic therapies. Therefore, biofilms are major contributors to the global health problem of antibiotic resistance, and novel approaches to counter them are urgently needed. Small molecules of the innate immune system called host defense peptides (HDPs) have emerged as promising templates for the design of potent, broad-spectrum antibiofilm agents. Here, we review recent developments in the new field of synthetic antibiofilm peptides, including mechanistic insights, synergistic interactions with available antibiotics, and their potential as novel antimicrobials against persistent infections caused by biofilms. This article is part of a Special Issue entitled: Antimicrobial peptides edited by Karl Lohner and Kai Hilpert. PMID:26724202

  19. Inexpensive Neutron Imaging Cameras Using CCDs for Astronomy

    NASA Astrophysics Data System (ADS)

    Hewat, A. W.

    We have developed inexpensive neutron imaging cameras using CCDs originally designed for amateur astronomical observation. The low-light, high resolution requirements of such CCDs are similar to those for neutron imaging, except that noise as well as cost is reduced by using slower read-out electronics. For example, we use the same 2048x2048 pixel "Kodak" KAI-4022 CCD as used in the high performance PCO-2000 CCD camera, but our electronics requires ∼5 sec for full-frame read-out, ten times slower than the PCO-2000. Since neutron exposures also require several seconds, this is not seen as a serious disadvantage for many applications. If higher frame rates are needed, the CCD unit on our camera can be easily swapped for a faster readout detector with similar chip size and resolution, such as the PCO-2000 or the sCMOS PCO.edge 4.2.

  20. Feature

    NASA Astrophysics Data System (ADS)

    2011-01-01

    Shi Changxu, former vice-president of NSFC wins Top Prize of National Science and Technology Award of China Both China and the world materials community have greatly benefitted from his service, by RPH Chang Shi Changxu—a great teacher and mentor for materials scientists, by Gaoqing Max Lu A bright example for all of us—Professor Shi Changxu, by Wei Gao Professor Shi Changxu—The Giant Materials Scientist of China, by Wuzong Zhou Congratulations to Academician Changxu Shi on the Occasion of His Winning the 2010 Chinese Science & Technology Grand Prize, by Ju Li, Kai Chen, Zhiwei Shan, Guanjun Qiao, Jun Sun and Evan Ma Materials—the foundation for technology revolutions, by Zhong Lin Wang

  1. Inhibition of VEGF-dependent angiogenesis by the anti-CD82 monoclonal antibody 4F9 through regulation of lipid raft microdomains.

    PubMed

    Nomura, Sayaka; Iwata, Satoshi; Hatano, Ryo; Komiya, Eriko; Dang, Nam H; Iwao, Noriaki; Ohnuma, Kei; Morimoto, Chikao

    2016-05-20

    CD82 (also known as KAI1) belongs to the tetraspanin superfamily of type III transmembrane proteins, and is involved in regulating cell adhesion, migration and proliferation. In contrast to these well-established roles of CD82 in tumor biology, its function in endothelial cell (EC) activity and tumor angiogenesis is yet to be determined. In this study, we show that suppression of CD82 negatively regulates vascular endothelial growth factor (VEGF)-induced angiogenesis. Moreover, we demonstrate that the anti-CD82 mAb 4F9 effectively inhibits phosphorylation of VEGF receptor 2 (VEGFR2), which is the principal mediator of the VEGF-induced angiogenic signaling process in tumor angiogenesis, by regulating the organization of the lipid raft microdomain signaling platform in human EC. Our present work therefore suggests that CD82 on EC is a potential target for anti-angiogenic therapy in VEGFR2-dependent tumor angiogenesis. PMID:27103437

  2. Proton induced tautomeric switching in N-rich aromatics with tunable acid-base character

    NASA Astrophysics Data System (ADS)

    Centore, Roberto; Manfredi, Carla; Fusco, Sandra; Maglione, Cira; Carella, Antonio; Capobianco, Amedeo; Peluso, Andrea; Colonna, Daniele; Di Carlo, Aldo

    2015-08-01

    The acid-base properties of selected derivatives of the [1,2,4]triazolo[3,2-c][1,2,4]triazole fused aromatic system have been investigated by UV-vis spectroscopy. Neutral heterobicycles (HL) exhibit amphoteric behavior (they can deliver the N-H proton forming the conjugated base L- and can accept up to two protons, forming the species H2L+ and H3L++) and show an unprecedented tautomeric switching upon protonation, as revealed by single crystal X-ray analysis and confirmed by theoretical calculations. By varying the groups attached at the heterocycle, a remarkable shift of pKai values, up to 5-6 units, is observed. In particular, with strong electron attractor groups at position 7 (e.g. p-nitrophenyl or pentafluorophenyl) the neutral compounds are stronger acids than phenol or p-nitrophenol.

  3. Noise in Nonlinear Dynamical Systems

    NASA Astrophysics Data System (ADS)

    Moss, Frank; McClintock, P. V. E.

    2009-08-01

    List of contributors; Preface; Introduction to volume three; 1. The effects of coloured quadratic noise on a turbulent transition in liquid He II J. T. Tough; 2. Electrohydrodynamic instability of nematic liquid crystals: growth process and influence of noise S. Kai; 3. Suppression of electrohydrodynamic instabilities by external noise Helmut R. Brand; 4. Coloured noise in dye laser fluctuations R. Roy, A. W. Yu and S. Zhu; 5. Noisy dynamics in optically bistable systems E. Arimondo, D. Hennequin and P. Glorieux; 6. Use of an electronic model as a guideline in experiments on transient optical bistability W. Lange; 7. Computer experiments in nonlinear stochastic physics Riccardo Mannella; 8. Analogue simulations of stochastic processes by means of minimum component electronic devices Leone Fronzoni; 9. Analogue techniques for the study of problems in stochastic nonlinear dynamics P. V. E. McClintock and Frank Moss; Index.

  4. Controlling bacterial infections by inhibiting proton-dependent processes.

    PubMed

    Kaneti, Galoz; Meir, Ohad; Mor, Amram

    2016-05-01

    Bacterial resistance to antibiotics is recognized as one of the greatest threats in modern healthcare, taking a staggering toll worldwide. New approaches for controlling bacterial infections must be designed, eventually combining multiple strategies for complimentary therapies. This review explores an old/new paradigm for multi-targeted antibacterial therapy, focused at disturbing bacterial cytoplasmic membrane functions at sub minimal inhibitory concentrations, namely through superficial physical alterations of the bilayer, thereby perturbing transmembrane signals transduction. Such a paradigm may have the advantage of fighting the infection while avoiding many of the known resistance mechanisms. This article is part of a Special Issue entitled: Antimicrobial peptides edited by Karl Lohner and Kai Hilpert. PMID:26522076

  5. Reptin and Pontin function antagonistically with PcG and TrxG complexes to mediate Hox gene control

    PubMed Central

    Diop, Soda Balla; Bertaux, Karine; Vasanthi, Dasari; Sarkeshik, Ali; Goirand, Benjamin; Aragnol, Denise; Tolwinski, Nicholas S; Cole, Michael D; Pradel, Jacques; Yates, John R; Mishra, Rakesh K; Graba, Yacine; Saurin, Andrew J

    2008-01-01

    Pontin (Pont) and Reptin (Rept) are paralogous ATPases that are evolutionarily conserved from yeast to human. They are recruited in multiprotein complexes that function in various aspects of DNA metabolism. They are essential for viability and have antagonistic roles in tissue growth, cell signalling and regulation of the tumour metastasis suppressor gene, KAI1, indicating that the balance of Pont and Rept regulates epigenetic programmes critical for development and cancer progression. Here, we describe Pont and Rept as antagonistic mediators of Drosophila Hox gene transcription, functioning with Polycomb group (PcG) and Trithorax group proteins to maintain correct patterns of expression. We show that Rept is a component of the PRC1 PcG complex, whereas Pont purifies with the Brahma complex. Furthermore, the enzymatic functions of Rept and Pont are indispensable for maintaining Hox gene expression states, highlighting the importance of these two antagonistic factors in transcriptional output. PMID:18259215

  6. A spectroscopic study of phenylbutazone and aspirin bound to serum albumin in rheumatoid diseases

    NASA Astrophysics Data System (ADS)

    Maciążek-Jurczyk, M.; Sułkowska, A.; Bojko, B.; Równicka-Zubik, J.; Sułkowski, W. W.

    2011-11-01

    Interaction of phenylbutazone (PBZ) and aspirin (ASA), two drugs recommended in rheumatoid diseases (RDs), when binding to human (HSA) and bovine (BSA) serum albumins, has been studied by quenching of fluorescence and proton nuclear magnetic resonance ( 1HNMR) techniques. On the basis of spectrofluorescence measurements high affinity binding sites of PBZ and ASA on albumin as well as their interaction within the binding sites were described. A low affinity binding site has been studied by proton nuclear magnetic resonance spectroscopy. Using fluorescence spectroscopy the location of binding site in serum albumin (SA) for PBZ and ASA was found. Association constants Ka were determined for binary (i.e. PBZ-SA and ASA-SA) and ternary complexes (i.e. PBZ-[ASA]-SA and ASA-[PBZ]-SA). PBZ and ASA change the affinity of each other to the binding site in serum albumin (SA). The presence of ASA causes the increase of association constants KaI of PBZ-SA complex. Similarly, PBZ influences KaI of ASA-SA complex. This phenomenon shows that the strength of binding and the stability of the complexes increase in the presence of the second drug. The decrease of KaII values suggests that the competition between PBZ and ASA in binding to serum albumin in the second class of binding sites occurs. The analysis of 1HNMR spectral parameters i.e. changes of chemical shifts and relaxation times of the drug indicate that the presence of ASA weakens the interaction of PBZ with albumin. Similarly PBZ weakens the interaction of ASA with albumin. This conclusion points to the necessity of using a monitoring therapy owning to the possible increase of uncontrolled toxic effects.

  7. Generic temperature compensation of biological clocks by autonomous regulation of catalyst concentration

    PubMed Central

    Hatakeyama, Tetsuhiro S.; Kaneko, Kunihiko

    2012-01-01

    Circadian clocks—ubiquitous in life forms ranging from bacteria to multicellular organisms—often exhibit intrinsic temperature compensation; the period of circadian oscillators is maintained constant over a range of physiological temperatures, despite the expected Arrhenius form for the reaction coefficient. Observations have shown that the amplitude of the oscillation depends on the temperature but the period does not; this suggests that although not every reaction step is temperature independent, the total system comprising several reactions still exhibits compensation. Here we present a general mechanism for such temperature compensation. Consider a system with multiple activation energy barriers for reactions, with a common enzyme shared across several reaction steps. The steps with the highest activation energy rate-limit the cycle when the temperature is not high. If the total abundance of the enzyme is limited, the amount of free enzyme available to catalyze a specific reaction decreases as more substrates bind to the common enzyme. We show that this change in free enzyme abundance compensates for the Arrhenius-type temperature dependence of the reaction coefficient. Taking the example of circadian clocks with cyanobacterial proteins KaiABC, consisting of several phosphorylation sites, we show that this temperature compensation mechanism is indeed valid. Specifically, if the activation energy for phosphorylation is larger than that for dephosphorylation, competition for KaiA shared among the phosphorylation reactions leads to temperature compensation. Moreover, taking a simpler model, we demonstrate the generality of the proposed compensation mechanism, suggesting relevance not only to circadian clocks but to other (bio)chemical oscillators as well. PMID:22566655

  8. Prostate-derived ets factor represses tumorigenesis and modulates epithelial-to-mesenchymal transition in bladder carcinoma cells.

    PubMed

    Tsui, Ke-Hung; Lin, Yu-Hsiang; Chung, Li-Chuan; Chuang, Sung-Ting; Feng, Tsui-Hsia; Chiang, Kun-Chun; Chang, Phei-Lang; Yeh, Chi-Ju; Juang, Horng-Heng

    2016-05-28

    Prostate-derived Ets (E-twenty six) factor (PDEF), an epithelium-specific member of the Ets family of transcription factors, has been shown to play a role in suppressing the development of many epithelium-derived cancers such as prostate and breast cancer. It is not clear, however, whether PDEF is involved in the development or progression of bladder cancer. In a comparison between normal urothelium and bladder tumor tissue, we identified significant decreases of PDEF in the tumor tissue. Further, the immunohistochemistry assays indicated a significantly higher immunostaining of PDEF in low-grade bladder tumors. Additionally, the highly differentiated transitional-cell bladder carcinoma RT-4 cells expressed significantly more PDEF levels than the bladder carcinoma HT1376 and the T24 cells. Ectopic overexpression of PDEF attenuated proliferation, invasion, and tumorigenesis of bladder carcinoma cells in vitro and in vivo. PDEF enhanced the expression levels of mammary serine protease inhibitor (MASPIN), N-myc downstream regulated gene 1 (NDRG1), KAI1, and B-cell translocation gene 2 (BTG2). PDEF modulated epithelial-mesenchymal-transition (EMT) by upregulating E-cadherin expression and downregulating the expression of N-cadherin, SNAIL, SLUG, and vimentin, leading to lower migration and invasion abilities of bladder carcinoma cells. Filamentous actin (F-actin) polarization and remodeling were observed in PDEF-knockdown RT-4 cells. Our results suggest that PDEF gene expression is associated with the extent of bladder neoplasia and PDEF modulated the expressions of EMT-related genes. The induction of BTG2, NDRG1, MASPIN, and KAI1 gene expressions by PDEF may explain the inhibitory functions of PDEF on the proliferation, invasion, and tumorigenesis in bladder carcinoma cells. PMID:26965996

  9. Orphans and Vulnerable Children in Kenya: Results From a Nationally Representative Population-Based Survey

    PubMed Central

    Lee, Veronica C.; Muriithi, Patrick; Gilbert-Nandra, Ulrike; Kim, Andrea A.; Schmitz, Mary E.; Odek, James; Mokaya, Rose; Galbraith, Jennifer S.

    2016-01-01

    Background In Kenya, it is estimated that there are approximately 3.6 million children aged <18 years who have been orphaned or who are vulnerable. We examined the data from the second Kenya AIDS Indicator Survey (KAIS 2012) to determine the number and profile of orphans and vulnerable children (OVC) in Kenya who were aged <18 years. Methods KAIS 2012 was a nationally representative, population-based household survey. We analyzed the data for all the children from birth to age 17 years who resided in an eligible household so as to determine whether their parents were alive or had been very ill to define their OVC status. Results We estimated that there were 2.6 million OVC in Kenya in 2012, of whom 1.8 million were orphans and 750,000 were vulnerable. Among orphans, 15% were double orphans. Over one-third of all the OVC were aged between 10 and 14 years. Households with ≥1 OVC (12% of all households) were usually in the lowest 2 wealth quintiles, and 22% of OVC households had experienced moderate or severe hunger. Receipt of OVC support services was low for medical (3.7%), psychological (4.1%), social (1.3%), and material support (6.2%); educational support was slightly more common (11.5%). Orphanhood among children aged <15 years increased from 1993 to 2003 (P < 0.01) but declined from 2003 to 2012 (P < 0.01). Conclusions The 2.6 million OVC constitute a significant proportion of Kenya’s population aged <18 years. Special attention should be paid to OVC to prevent further vulnerability and ensure their well-being and development as they transition into adulthood. PMID:24732824

  10. Burden of HIV Infection Among Children Aged 18 Months to 14 Years in Kenya: Results From a Nationally Representative Population-Based Cross-sectional Survey

    PubMed Central

    Ng’eno, Bernadette; Mwangi, Ann; Ng’ang’a, Lucy; Kim, Andrea A.; Waruru, Anthony; Mukui, Irene; Ngugi, Evelyn W.; Rutherford, George W.

    2016-01-01

    Backgrounds In Kenya, mathematical models estimate that there are approximately 220,000 children aged less than 15 years infected with HIV. We analyzed data from the second Kenya AIDS Indicator Survey (KAIS 2012) to estimate the prevalence of HIV infection among children aged 18 months to 14 years. Methods KAIS 2012 was a nationally representative 2-stage cluster sample household survey. We studied children aged 18 months to 14 years whose parents or guardians answered questions pertaining to their children by interview. Blood specimens were collected for HIV serology and viral load measurement. Results We identified 5162 children who were eligible for the study. Blood was obtained for 3681 (71.3%) children. Among child participants, 16.4% had been tested for HIV infection in the past, and among children with parents or guardians who self-reported HIV-positive status, 52.9% had been tested for HIV infection. Twenty-eight (0.9%) children tested HIV-positive in the survey. Of these, 11 had been previously diagnosed with HIV infection before the survey. All 11 children were in HIV care and receiving cotrimoxazole; 8 were on antiretorivral therapy (ART). Among those on ART, 4 were virologically suppressed. Conclusions HIV causes a substantial burden of disease in the Kenyan pediatric population. Although most children who had been diagnosed with HIV before the survey were engaged in care and treatment, they represented less than half of HIV-infected children identified in the survey. Future efforts should focus on identifying infected children and getting them into care and on suppressive ART as early as possible. PMID:24732823

  11. Status of Voluntary Medical Male Circumcision in Kenya: Findings From 2 Nationally Representative Surveys in Kenya, 2007 and 2012

    PubMed Central

    Galbraith, Jennifer S.; Ochieng, Athanasius; Mwalili, Samuel; Emusu, Donath; Mwandi, Zebedee; Kim, Andrea A.; Rutherford, George; Maina, William K.; Kimanga, Davies O.; Chesang, Kipruto; Cherutich, Peter

    2016-01-01

    Background The Kenyan Ministry of Health initiated a voluntary medical male circumcision (VMMC) program in 2008. We used data from 2 nationally representative surveys to estimate trends in the number, demographic characteristics, and sexual behaviors of recently circumcised and uncircumcised HIV-uninfected men in Kenya. Methods We compared the proportion of circumcised men between the first and second Kenya AIDS Indicator Survey (KAIS 2007 and KAIS 2012) to assess the progress of Kenya’s VMMC program. We calculated the number of uncircumcised HIV-uninfected men. We conducted descriptive analyses and used multivariable methods to identify the variables independently associated with HIV-uninfected uncircumcised men aged 15–64 years in the VMMC priority region of Nyanza. Results The proportion of men who reported being circumcised increased significantly from 85.0% in 2007 to 91.2% in 2012. The proportions of circumcised men increased in all regions, with the highest increases of 18.1 and 9.0 percentage points in the VMMC priority regions of Nyanza and Nairobi, respectively. Half (52.5%) of HIV-uninfected and uncircumcised men had never been married, and 84.6% were not using condoms at all times with their last sexual partner. Conclusions VMMC prevalence has increased across Kenya demonstrating the success of the national program. Despite this accomplishment, the Nyanza region remains below the target to circumcise 80% of all eligible men aged 15–49 years between 2009 and 2013. As new cohorts of young men enter into adolescence, consistent focus is needed. To ensure sustainability of the VMMC program, financial resources and coordinated planning must continue. PMID:24732820

  12. A spectroscopic study of phenylbutazone and aspirin bound to serum albumin in rheumatoid diseases.

    PubMed

    Maciążek-Jurczyk, M; Sułkowska, A; Bojko, B; Równicka-Zubik, J; Sułkowski, W W

    2011-11-01

    Interaction of phenylbutazone (PBZ) and aspirin (ASA), two drugs recommended in rheumatoid diseases (RDs), when binding to human (HSA) and bovine (BSA) serum albumins, has been studied by quenching of fluorescence and proton nuclear magnetic resonance ((1)HNMR) techniques. On the basis of spectrofluorescence measurements high affinity binding sites of PBZ and ASA on albumin as well as their interaction within the binding sites were described. A low affinity binding site has been studied by proton nuclear magnetic resonance spectroscopy. Using fluorescence spectroscopy the location of binding site in serum albumin (SA) for PBZ and ASA was found. Association constants K(a) were determined for binary (i.e. PBZ-SA and ASA-SA) and ternary complexes (i.e. PBZ-[ASA]-SA and ASA-[PBZ]-SA). PBZ and ASA change the affinity of each other to the binding site in serum albumin (SA). The presence of ASA causes the increase of association constants K(aI) of PBZ-SA complex. Similarly, PBZ influences K(aI) of ASA-SA complex. This phenomenon shows that the strength of binding and the stability of the complexes increase in the presence of the second drug. The decrease of K(aII) values suggests that the competition between PBZ and ASA in binding to serum albumin in the second class of binding sites occurs. The analysis of (1)HNMR spectral parameters i.e. changes of chemical shifts and relaxation times of the drug indicate that the presence of ASA weakens the interaction of PBZ with albumin. Similarly PBZ weakens the interaction of ASA with albumin. This conclusion points to the necessity of using a monitoring therapy owning to the possible increase of uncontrolled toxic effects. PMID:21856214

  13. Biomonitoring of heavy metals and trace organics using the intertidal mussel Perna viridis in Hong Kong coastal waters.

    PubMed

    Liu, J H; Kueh, C S W

    2005-01-01

    This paper presents the results of a 6-year (1998-2003) survey of trace toxics in the intertidal mussel Perna viridis conducted by the Hong Kong Environmental Protection Department. Concentrations of heavy metals and trace organics were measured in the soft bodies of P. viridis collected from five sites in Hong Kong waters, i.e. Wu Kai Sha (Tolo Harbour), Ma Wan (Northwest), Tsim Sha Tsui (Victoria Harbour), Tai Tam (Hong Kong South) and Lamma Island (Southern Waters) in order to establish the spatial patterns of contaminants in mussels. Among the metals analysed, Cd showed a significant concentration gradient in Hong Kong waters. The levels of Cd in P. viridis were significantly higher at Ma Wan as compared to the other sites studied. Ma Wan also had relatively higher concentrations of Pb. Mn concentrations were particularly prominent at Wu Kai Sha. Significantly higher concentrations of Hg and Cu were recorded at Tai Tam and Tsim Sha Tsui. Tai Tam and Wu Kai Sha had higher levels of V; whereas higher Ni concentrations were recorded at Lamma Island and Tai Tam. No clear spatial patterns for Al, As, Cr, Fe and Zn were observed. Higher concentrations of PAHs in P. viridis were observed around urban centres impacted by sewage discharges (e.g. Tsim Sha Tsui); whereas higher PCB levels were found not only in Tsim Sha Tsui but also in less urbanised areas such as Lamma Island and Tai Tam, suggesting that these may be due to non-sewage related inputs. The study also shows that Northwest and Southern waters are subject to a higher degree of DDT pollution compared with other sites. Of the 17 dioxin compounds analysed, positive data were mostly recorded for two compounds which are of low toxicity (i.e. OCDD and 1,2,3,4,6,7,8-HpCDD) whereas the most toxic congeners (i.e. 2,3,7,8-TCDD and 1,2,3,7,8-PeCDD) were not detected in the 6 years of monitoring. In general, the levels of OCDD in P. viridis were found to be higher in Tai Tam and Lamma Island in Southern Waters of Hong

  14. Links between two different types of spectra of charged nanometer aerosol particles

    NASA Astrophysics Data System (ADS)

    Luts, A.; Komsaare, K.; Parts, T.-E.; Hõrrak, U.

    2011-08-01

    We have, since 2007, continuously measured the electrical mobility distribution of small (< 1.5 nm in diameter) corona-generated one-second-aged air ions, using our Small Air Ion Spectrometer (KAIS), in urban area, in the center of the town of Tartu, Estonia. We have simultaneously measured the mobility distributions of natural air ions (0.42-7.4 nm in diameter) with the Balanced Scanning Mobility Analyzer (BSMA). In this work we employ these data to establish certain links between the concurrent spectra of two types, especially for days with new particle formation events. We elaborated and tested an automatic classification method, which selected the spectra according their shape. In the case of the BSMA negative ions, we obtained four classes of the spectra which are associated with the intermediate ion (1.6-7.4 nm in diameter) nucleation burst events and rain-type events. The spectra within these classes are characterized by special shapes and they are called "event-like spectra". The first class demonstrates the strongest increase in the concentration of all ions with the mobilities below 0.8 cm 2V -1 s -1 (above 1.25 nm in diameter), it mainly contains spring and early summer spectra, recorded around midday. In the second class, the events are weaker; it contains a large number of late spring morning time spectra, when relative humidity (RH) tends to decrease. The third and the fourth classes contain many spectra, which resemble to short isolated events. These spectra are excluded from the further analysis. In the first class, the precipitation spectra make about 34% of all the event-like spectra, in the other classes about 10%. In the case of the BSMA positive ions, only the first and second classes were present. The precipitation spectra make about 20% of all event-like classes. About 45% of the event-like spectra are without precipitation, but with RH > 90%, and these spectra are excluded from the further analysis. The spectra with RH < 50% formed 12% of

  15. The footprints of typhoons on seismic records and their implications on small-scale coupling mechanisms in South China Sea

    NASA Astrophysics Data System (ADS)

    Xiao, H.; Xue, M.; Yang, T.; Liu, C.; Hua, Q.; Xia, S.; Huang, H.; Le, B. M.; Huo, D.; Pan, M.; Li, L.

    2015-12-01

    By investigating the footprints of typhoons on seismic records, we can understand their contributions to seismic noises as well as to small-scale coupling mechanisms of typhoon-land and typhoon-ocean-land. We analyze the signatures of typhoon KAI-TAK and BOPHA using seismic data from the ocean-bottom seismometers (OBSs) deployed in the central basin of South China Sea by Tongji University in 2012 as well as seismic stations (IC.QIZ,HK.HKPS and RM.SZP) on lands.Our preliminary results show that typhoons mainly enhance microseisms at the frequency band of ~0.1-0.5 Hz, including both long period double frequency (LPDF) and short period double frequency (SPDF) microseisms. A positive correlation observed between the amplitude of microseisms and the height of local ocean waves. Because OBSs are deployed at the bottom of ocean, single frequency (SF) microseisms are not prominent on them due to their fast attenuation with depth. During the typhoon KAI-TAK, the increase of LPDF energy is very small in OBSs while that is very high on land stations, indicating that LPDF microseisms are generated at nearby shorelines and can propagate towards the sea through the solid earth. However, the increase of SPDF energy is almost the same level for both OBSs and land stations indicating that the generation of SPDF is probably local.However, we also observe a small amount of energy arrives before the increases of the wave heights at the land station HK.HKPS. We derive that this energy may from a source that is not local: while LPDF can be generated at nearby shorelines and SPDF can be generated everywhere locally, they can both transmit through the solid part of the Earth to a station some distance away, i.e. HK.HKPS. In addition, we find that typhoons enhance not only the microseisms as expected but also the seismic energy from higher frequency bands. The spectrum amplitude during Typhoon periods, normalized by that of no-storm periods, shows that land stations produce stronger higher

  16. Organochlorines and heavy metals in wild caught food as a potential human health risk to the indigenous Māori population of South Canterbury, New Zealand.

    PubMed

    Stewart, Michael; Phillips, Ngaire R; Olsen, Greg; Hickey, Christopher W; Tipa, Gail

    2011-05-01

    Increasing concentrations of anthropogenic contaminants in wild kai (food) of cultural, recreational and economic importance to the indigenous Māori of New Zealand is a potential human health risk. Contaminants that are known to bioaccumulate through the food chain (e.g., organochlorine pesticides (OCPs), PCBs and selected heavy metals) were analysed in important kai species including eel (Anguilla sp.), brown trout (Salmo trutta), black flounder (Rhombosolea retiaria) and watercress (Nasturtium officinale) from important harvesting sites in the region of South Canterbury. Eels contained relatively high wet weight concentrations of p,p'-DDE (8.6-287ng/g), PCBs ((32)Σ(PCB); 0.53-58.3ng/g), dieldrin (<0.05-16.3ng/g) and Σchlordanes (0.03-10.6ng/g). Trout and flounder contained lower concentrations of organochlorines than eels, with p,p'-DDE wet weight concentrations ranging from 2.2 to 18.5ng/g for trout and 6.4 to 27.8ng/g for flounder. Total arsenic wet weight concentrations were below detection limits for eels but ranged from 0.27 to 0.89μg/g for trout and 0.12 to 0.56μg/g for flounder. Mercury concentrations ranged from 0.02 to 0.56μg/g, 0.11 to 0.50μg/g and 0.04 to 0.10μg/g (ww) for eel, trout and flounder respectively. Lifetime excess cancer risk was calculated through established risk assessment procedures, highlighting dieldrin, ΣPCBs and p,p'-DDE in eels and arsenic in trout and flounder as primary contaminants of concern. A second non-cancer chronic health risk assessment indicated that mercury and PCBs were a potential concern in eels and mercury in trout. A cumulative lifetime cancer risk assessment showed potential health risk for consumption of some species, even at low consumption rates and provided the basis for establishing recommended dietary consumption limits for harvest sites within the study region. PMID:21402398

  17. [Experiences in introducing and using a patient data management system (PDMS). in anesthesiology at the Clinic of Anesthesiology and Intensive Care Medicine of the University of Leipzig].

    PubMed

    Friese, St; Olthoff, D

    2003-01-01

    In the field of anaesthesia the demands on the quality and quantity of documentation are increasing constantly. Patient Data Management Systems (PDMS) have proved an effective means of handling the volume of data generated. The main reasons for introducing a PDMS vary greatly, nevertheless, it is possible to formulate general requirements such as those of the "Position Paper of the Study Group on Patient Data Management Systems (PDMS) of the University Departments of Anaesthesiology in Bavaria". Although these requirements are very broad, they provide a good basis for comparing different approaches to computer-assisted documentation in anaesthesiology. The stage currently reached at the Department of Anaesthesiology and Intensive Care Medicine (KAI) of the University of Leipzig is analysed in comparison with the position paper. The COPRA system was established at KAI Leipzig eight years ago. It was developed from an existing version for intensive care medicine. It meets the demands made on it when it was introduced and can be enlarged and adjusted to the special needs of anaesthesiology. One particular requirement was that it should be possible to handle computer-assisted documentation and conventional documentation on paper simultaneously. This requirement is met by making the printed forms and those shown on the VDU practically the same in appearance. The anaesthetist is able to recognize "his" record on the screen. This greatly reduces the time required for familiarization and training. If possible, the orientation and updating of the system should be in the hands of an anaesthetist, since this is the only way to ensure that it remains geared primarily to medical needs. Administrative aspects have to be taken into account, but they should not dominate the system. The anaesthetist managing the system should have some basic training in EDP, or at least take a special interest in it. This ensures that minor enlargements can be carried out easily, as soon as

  18. Objective Identification of Antarctic Katabatic Outflow Events On Different Spatial and Temporal Scales

    NASA Astrophysics Data System (ADS)

    Spangehl, T.; Leckebusch, G. C.; Speth, P.

    Katabatic winds are a significant climatological feature of the boundary layer over Antarctica. These drainage flow systems of cold air masses are steered by orography and often converge in several coastal regions around Antarctica where they lead to strong and persistent katabatic winds. The aim of this study is to investigate the rela- tionship between outbreaks of cold air masses over Antarctica and anomalies of the Southern Hemisphere atmospheric circulation. At first we will therefore focus on the objective identification of katabatic outbreak events. The principal idea is to define an index, which we called KAI (from German: Katabatischer Abfluß Index), as indicator for katabatic outflow. First data of automatic weather stations (provided by University of Wisconsin) and GTS-Synops (British Antarctic Survey) are used to define a regional katabatic outflow index. Second, based on operational (ECMWF-) analysis a continen- tal scale outflow index is constructed by means of mass flow. Whereas the definition of the regional indices is only based on wind data further meteorological parameters as for example temperature or the strength of the surface inversion are taken into account, to exclude events that are of pure synoptic nature and are not of katabatic origin. Time series of KAI are analysed with regard to the variabilty of the identified outflow events focussing on strength and duration. Regional indices are compared against each other to investigate larger scale connections of katabatic outflow. It is especially tested if the continental scale index can be compared to the regional indices in terms of variability. Additionally, by the aid of backward trajectories, different regimes of katabatic wind systems can be recognised. So a classification of katabatic outflow events is achieved. Finally, index based composite studies of the geopotential height are performed to gain a first look insight the connection to the hemispheric circulation. It can be shown that

  19. Lack of Knowledge of HIV Status a Major Barrier to HIV Prevention, Care and Treatment Efforts in Kenya: Results from a Nationally Representative Study

    PubMed Central

    Cherutich, Peter; Kaiser, Reinhard; Galbraith, Jennifer; Williamson, John; Shiraishi, Ray W.; Ngare, Carol; Mermin, Jonathan; Marum, Elizabeth; Bunnell, Rebecca

    2012-01-01

    Background We analyzed HIV testing rates, prevalence of undiagnosed HIV, and predictors of testing in the Kenya AIDS Indicator Survey (KAIS) 2007. Methods KAIS was a nationally representative sero-survey that included demographic and behavioral indicators and testing for HIV, HSV-2, syphilis, and CD4 cell counts in the population aged 15–64 years. We used gender-specific multivariable regression models to identify factors independently associated with HIV testing in sexually active persons. Results Of 19,840 eligible persons, 80% consented to interviews and blood specimen collection. National HIV prevalence was 7.1% (95% CI 6.5–7.7). Among ever sexually active persons, 27.4% (95% CI 25.6–29.2) of men and 44.2% (95% CI 42.5–46.0) of women reported previous HIV testing. Among HIV-infected persons, 83.6% (95% CI 76.2–91.0) were unaware of their HIV infection. Among sexually active women aged 15–49 years, 48.7% (95% CI 46.8–50.6) had their last HIV test during antenatal care (ANC). In multivariable analyses, the adjusted odds ratio (AOR) for ever HIV testing in women ≥35 versus 15–19 years was 0.2 (95% CI: 0.1–0.3; p<0.0001). Other independent associations with ever HIV testing included urban residence (AOR 1.6, 95% CI: 1.2–2.0; p = 0.0005, women only), highest wealth index versus the four lower quintiles combined (AOR 1.8, 95% CI: 1.3–2.5; p = 0.0006, men only), and an increasing testing trend with higher levels of education. Missed opportunities for testing were identified during general or pregnancy-specific contacts with health facilities; 89% of adults said they would participate in home-based HIV testing. Conclusions The vast majority of HIV-infected persons in Kenya are unaware of their HIV status, posing a major barrier to HIV prevention, care and treatment efforts. New approaches to HIV testing provision and education, including home-based testing, may increase coverage. Targeted interventions should involve sexually active men

  20. Antiretroviral Treatment Scale-up Among Persons Living With HIV in Kenya: Results From a Nationally Representative Survey

    PubMed Central

    Odhiambo, Jacob O.; Kellogg, Timothy A.; Kim, Andrea A.; Ng’ang’a, Lucy; Mukui, Irene; Umuro, Mamo; Mohammed, Ibrahim; De Cock, Kevin M.; Kimanga, Davies O.; Schwarcz, Sandra

    2016-01-01

    Background In 2007, 29% of HIV-infected Kenyans in need of antiretroviral therapy (ART), based on an immunologic criterion of CD4 ≤350 cells per microliter, were receiving ART. Since then, substantial treatment scale-up has occurred in the country. We analyzed data from the second Kenya AIDS Indicator Survey (KAIS 2012) to assess progress of treatment scale-up in Kenya. Methods KAIS 2012 was a nationally representative survey of persons aged 18 months to 64 years that collected information on HIV status, care, and treatment. ART eligibility was defined based on 2 standards: (1) 2011 Kenya eligibility criteria for ART initiation: CD4 ≤350 cells per microliter or co-infection with active tuberculosis and (2) 2013 World Health Organization (WHO) eligibility criteria for ART initiation: CD4 ≤500 cells per microliter, co-infection with active tuberculosis, currently pregnant or breastfeeding, and infected partners in serodiscordant relationships. Blood specimens were tested for HIV antibodies and HIV-positive specimens tested for CD4 cell counts. Results Among 13,720 adults and adolescents aged 15–64 years, 11,626 provided a blood sample, and 648 were HIV infected. Overall, 58.8% [95% confidence interval (CI): 52.0 to 65.5) were eligible for treatment using the 2011 Kenya eligibility criteria and 77.4% (95% CI: 72.4 to 82.4) using the 2013 WHO eligibility criteria. Coverage of ART was 60.5% (95% CI: 50.8 to 70.2) using the 2011 Kenya eligibility criteria and 45.9% (95% CI: 37.7 to 54.2) using the 2013 WHO eligibility criteria. Conclusions ART coverage has increased from 29% in 2007 to 61% in 2012. If Kenya adopts the 2013 WHO guidelines for ART initiation, need for ART increases by an additional 19 percentage points and current coverage decreases by an additional 15 percentage points, representing an additional 214,000 persons who will need to be reached. PMID:24732815

  1. Factors Associated with HIV Infection in Married or Cohabitating Couples in Kenya: Results from a Nationally Representative Study

    PubMed Central

    Kaiser, Reinhard; Bunnell, Rebecca; Hightower, Allen; Kim, Andrea A.; Cherutich, Peter; Mwangi, Mary; Oluoch, Tom; Dadabhai, Sufia; Mureithi, Patrick; Mugo, Nelly; Mermin, Jonathan

    2011-01-01

    Background In order to inform prevention programming, we analyzed HIV discordance and concordance within couples in the Kenya AIDS Indicator Survey (KAIS) 2007. Methods KAIS was a nationally representative population-based sero-survey that examined demographic and behavioral indicators and serologic testing for HIV, HSV-2, syphilis, and CD4 cell counts in 15,853 consenting adults aged 15–64 years. We analyzed interview and blood testing data at the sexual partnership level from married or cohabitating couples. Multivariable regression models were used to identify factors independently associated with HIV discordant and concordant status. Results Of 3256 couples identified in the survey, 2748 (84.4%) had interview and blood testing data. Overall, 3.8% of couples were concordantly infected with HIV, and in 5.8% one partner was infected, translating to 338,000 discordant couples in Kenya. In 83.6% of HIV-infected Kenyans living in married or cohabitating couples neither partner knew their HIV status. Factors independently associated with HIV-discordance included young age in women (AOR 1.5, 95% CI: 1.2–1.8; p<0.0001), increasing number of lifetime sexual partners in women (AOR 1.5, 95% CI: 1.3–1.8; p<0.0001), HSV-2 infection in either or both partners (AOR 4.1, 95% CI: 2.3–7.2; p<0.0001), and lack of male circumcision (AOR 1.6, 95% CI: 1.0–2.5; p = 0.032). Independent factors for HIV-concordance included HSV-2 infection in both partners (AOR 6.5, 95% CI: 2.3–18.7; p = 0.001) and lack of male circumcision (AOR 1.8, 95% CI: 1.0–3.3; p = 0.043). Conclusions Couple prevention interventions should begin early in relationships and include mutual knowledge of HIV status, reduction of outside sexual partners, and promotion of male circumcision among HIV-uninfected men. Mechanisms for effective prevention or suppression of HSV-2 infection are also needed. PMID:21423615

  2. Engagement in HIV Care Among Kenyan Adults and Adolescents: Results From a National Population-Based Survey

    PubMed Central

    Wafula, Rose; Masyuko, Sarah; Ng’ang’a, Lucy; Kim, Andrea A.; Gichangi, Anthony; Mukui, Irene; Batuka, James; Ngugi, Evelyn W.; Maina, William K.; Schwarcz, Sandra

    2016-01-01

    Background Increasing access to care and treatment for HIV-infected persons is a goal in Kenya’s response to the HIV epidemic. Using data from the second Kenya AIDS Indicator Survey (KAIS 2012), we describe coverage of services received among adults and adolescents who were enrolled in HIV care. Methods KAIS 2012 was a population-based survey that collected information from persons aged 15–64 years that included self-reported HIV status, and for persons reporting HIV infection, use of HIV care and antiretroviral therapy (ART). Blood specimens were collected and tested for HIV. HIV-positive specimens were tested for CD4 counts and viral load. Results Among 363 persons who reported HIV infection, 93.4% [95% confidence interval (CI): 87.2 to 99.6] had ever received HIV care. Among those receiving HIV care, 96.3% (95% CI: 94.1 to 98.4) were using cotrimoxazole prophylaxis, and 74.6% (95% CI: 69.0 to 80.2) were receiving ART. A lower proportion of persons in care and not on ART reported using cotrimoxazole (89.5%, 95% CI: 82.5 to 96.5 compared with 98.6%, 95% CI: 97.1 to 100) and had a CD4 count measurement done (72.9%, 95% CI: 64.0 to 81.9 compared with 90.0%, 95% CI: 82.8 to 97.3) than persons in care and on ART, respectively. Among persons in care and not on ART, 23.2% (95% CI: 6.8 to 39.7) had CD4 counts ≤350 cells per microliter. Viral suppression was observed in 75.3% (95% CI: 68.7 to 81.9) of persons on ART. Conclusions Linkage and retention in care are high among persons with known HIV infection. However, improvements in care for the pre-ART population are needed. Viral suppression rates were comparable to developed settings. PMID:24732825

  3. Fluid Leakages in Gas-Hydrate Systems at the Storegga Slide of the Mid-Norwegian Margin

    NASA Astrophysics Data System (ADS)

    Hustoft, S.; Mienert, J.; Buenz, S.

    2005-12-01

    High-resolution 3D-seismic data has given new insight to the fluid flow regime at the well-documented gas-hydrate system along the northern flank of the Storegga Slide of the passive mid-Norwegian continental margin. Fluids are expelled from a widespread polygonal fault system of fine-grained, Miocene sediments of the lower Kai Formation that contribute to formation of gas hydrates in the study area. Previous work has mainly presumed a diffusive fluid flow regime above the polygonal fault system into the gas hydrate stability zone. However, 3D-seismic data indicates that migrating fluids above the polygonal fault system are focused trough imbricated small-offset fault zones in the upper stratigraphic level of the Kai Formation. Fluids seems to accumulate laterally within the lower level of the overlying Plio-Pleistocene Naust Formation, causing a small-scale dome structure with low frequency content, above the imbricated fault zones. Vertical fluid leakage originates from the extended dome structure causing fracturing in sediments throughout a ~200 m sedimentary sequence. The fluid leakage terminates within strata represented by negative amplitude anomalies located beneath a bottom-simulating reflector (BSR). Free gas in relatively porous media is deflected laterally by impermeable hydrated sediments occurring down-slope, and less permeable glacigenic sediments up-slope. At the upper termination of the fractured sediments within the Plio-Pleistocene formation, a 3 km long channel is recognized showing a geometric shape opposite to the stratigraphic deep-seated dome structure, but having the same lateral extension. The presence of vertical blowout pipes in direct proximity to the channel, indicate high pore pressure build-up. Adding to the complexity of the fluid flow system in gas hydrate settings, the channel might be a seismic artefact due to the presence of gas (pushdown), rather than a morphological feature. Due to the low organic matter content (< 1 %) of

  4. Geological evidence for historical and older earthquakes and tsunamis along the Nankai Trough, Japan

    NASA Astrophysics Data System (ADS)

    Garrett, Ed; De Batist, Marc; Heyvaert, Vanessa M. A.; Hubert-Ferrari, Aurélia; Fujiwara, Osamu; Yokoyama, Yusuke; Brückner, Helmut; Garrett, Philip

    2015-04-01

    In the wake of the devastating 2011 Tōhoku earthquake and tsunami, the Central Disaster Management Council of the Japanese Cabinet Office issued new guidance for assessing seismic hazards in Japan. Before 2011, seismic hazard assessment relied on source models developed from knowledge of a small number of well-documented historical earthquakes. Less well-known historical earthquakes, including the AD 869 Jōgan Sanriku earthquake, were largely disregarded as their seismic intensities or tsunami heights could not be reconciled with the chosen seismic sources. Following the unexpectedly large size of the Tōhoku earthquake, the Cabinet Office advocated renewed investigation of earthquake and tsunami occurrence over historical and longer timescales, with a particular focus on defining the largest possible magnitudes. The new guidelines pay close attention to the Nankai Trough, the subduction zone where the Philippine Sea Plate dives beneath the Eurasian Plate. The Nankai Trough faces the densely populated and highly industrialised coastline of south central Japan and harbours a widely-known seismic gap along its eastern Tōkai segment. A full-length rupture of the Nankai Trough, including the Tōkai segment, could produce an earthquake with a magnitude approaching that of the 2011 event, with tsunami travel times to the closest shorelines of less than 30 minutes. We review geological evidence for historical and older earthquakes and tsunamis along the Nankai Trough. This evidence comes from a wide variety of sources, including uplifted marine terraces, subsided marshes, liquefaction features, turbidites and tsunami deposits in coastal lakes and lowlands. Examining papers published before and after 2011, we investigate the impact of the new Cabinet Office guidelines on attempts to understand the magnitude and recurrence of these events. Additionally, we introduce the Belgian Science Policy Office funded QuakeRecNankai project, a collaboration aiming to supplement

  5. Neotectonic activity at the Giant Gjallar Vent (Norwegian Sea) indicates a future phase of active fluid venting

    NASA Astrophysics Data System (ADS)

    Dumke, Ines; Berndt, Christian; Crutchley, Gareth; Couillard, Mélanie; Gay, Aurélien

    2013-04-01

    The Giant Gjallar Vent (GGV) is a hydrothermal vent complex that formed during the opening of the North Atlantic at about 55 Ma. Sill intrusions into Cretaceous organic-rich sediments led to the production and subsequent vigorous seafloor venting of methane. A later phase of fluid escape occurred in mid-Oligocene times. The GGV is characterised by two pipes of 440 m and 480 m in diameter that reach up to the Base Late Pliocene Unconformity (BLPU) between the Kai and Naust formations. The unconformity is strongly deformed over an area of c. 18,000 km² across the vent, with a positive relief of up to 38 m above the surrounding paleo-seafloor. The overlying sediments of the Naust Formation conformally drape this deformation, smoothing its relief to a maximum of 15 m at the modern seafloor. The sediment drape indicates present inactivity of the vent system, as does the absence of indicators of active fluid escape in the water column during RV METEOR cruise M87-2 in 2012. However, high-resolution 2D seismic and Parasound data from the same cruise, and exploration-type 3D seismic data acquired by Norsk Hydro, show several indications for recent to ongoing activity at the GGV. Beneath the BLPU, strong frequency attenuation and chaotic reflections indicate the presence of free gas. At the edges of the extent of chaotic reflections, subvertical faults cut the unconformity as well as horizons of the lower and middle Naust Formation, suggesting tectonic activity after deposition of these horizons. Neotectonic activity is further indicated by the extensive occurrence of shallow faults apparent in Parasound records in the immediate vicinity of the vent and up to 16 km away. Some of these faults reach the seafloor. The observed deformation and faults may be the result of fluids accumulating beneath the BLPU due to increased loading of the oozy Kai Formation by denser glacigenic Naust sediments. Because of the lower permeability of the Naust Formation, the unconformity acts as a

  6. Ancient lenses in art and sculpture and the objects viewed through them, dating back 4500 years

    NASA Astrophysics Data System (ADS)

    Enoch, Jay M.

    1998-07-01

    The early history of lenses is controversial. The author has sought to address the problem by identifying lens elements (mainly convex/plano) which remain associated with objects intended to be viewed through them (i.e., in their original context). These are found in museums in sculptures, rings, pendants, etc. A number of outstanding examples will be illustrated in the talk; these sophisticated pieces of art are certainly not first constructs. Most are of rock crystal, rose quartz, or glass. Lenses have origin among artisans rather than scientists. Clearly, skills were often lost and rediscovered. Early lens-like objects have been found broadly in the eastern Mediterranean area/Middle East, in France, in Italy (Rome), and possibly in Peru and Scandinavia, etc. To date, the earliest lenses identified in context are from the IV/V Dynasties of Egypt, dating back to about 4500 years ago (e.g., the superb `Le Scribe Accroupi' and `the Kai' in the Louvre; added fine examples are located in the Cairo Museum). Latter examples have been found in Knossos (Minoan [Herakleion Museum]; ca. 3500 years ago); others had origin in Greece (examples in the Athens National Archeological Museum and the British Museum equals BM), in Rome (Metropolitan Museum, NY; BM; Vatican Museums; Bologna Archeological Museum), etc. Also. of great interest is the study of possible lens applications. This is a fascinating scientific, artistic and intellectual project.

  7. Global Transcriptomic Analysis Reveals the Mechanism of Phelipanche aegyptiaca Seed Germination.

    PubMed

    Yao, Zhaoqun; Tian, Fang; Cao, Xiaolei; Xu, Ying; Chen, Meixiu; Xiang, Benchun; Zhao, Sifeng

    2016-01-01

    Phelipanche aegyptiaca is one of the most destructive root parasitic plants of Orobanchaceae. This plant has significant impacts on crop yields worldwide. Conditioned and host root stimulants, in particular, strigolactones, are needed for unique seed germination. However, no extensive study on this phenomenon has been conducted because of insufficient genomic information. Deep RNA sequencing, including de novo assembly and functional annotation was performed on P. aegyptiaca germinating seeds. The assembled transcriptome was used to analyze transcriptional dynamics during seed germination. Key gene categories involved were identified. A total of 274,964 transcripts were determined, and 53,921 unigenes were annotated according to the NR, GO, COG, KOG, and KEGG databases. Overall, 5324 differentially expressed genes among dormant, conditioned, and GR24-treated seeds were identified. GO and KEGG enrichment analyses demonstrated numerous DEGs related to DNA, RNA, and protein repair and biosynthesis, as well as carbohydrate and energy metabolism. Moreover, ABA and ethylene were found to play important roles in this process. GR24 application resulted in dramatic changes in ABA and ethylene-associated genes. Fluridone, a carotenoid biosynthesis inhibitor, alone could induce P. aegyptiaca seed germination. In addition, conditioning was probably not the indispensable stage for P. aegyptiaca, because the transcript level variation of MAX2 and KAI2 genes (relate to strigolactone signaling) was not up-regulated by conditioning treatment. PMID:27428962

  8. Epstein-Barr virus-encoded small non-coding RNAs induce cancer cell chemoresistance and migration.

    PubMed

    Banerjee, Aditi Sengupta; Pal, Anindita Deb; Banerjee, Subrata

    2013-09-01

    Epstein-Barr virus (EBV) encoded small, non-coding, non-polyadenylated RNAs, known as EBERs are the most abundantly expressed viral transcripts in latently EBV infected cells. We found the specific role of EBERs in cell cycle progression, resistance against chemotherapeutic drug and cellular invasion in gastric cancer cells in vitro. Ectopic expression of EBERs upregulates the expression of IL-6 and activate its downstream STAT3, which is significantly involved in downregulating the expression of cell cycle inhibitor genes p21 and p27. Stable expression of EBERs regulates the activation of pFAK and pPAK1 and the expression of anti-metastatic genes RhoGDI and KAI-1 in gastric cancer cells. In addition, administration of neu-IL-6 antibody and dominant negative STAT3β reduces chemoresistance and inhibits invasion of EBERs-expressing gastric cancer cells. Our results thus revealed a novel role of EBERs in the coordination of IL-6-STAT3 signaling pathway to chemoresistance and cellular migration. PMID:23791019

  9. Determination of acrylamide in Thai-conventional snacks from Nong Mon market, Chonburi using GC-MS technique.

    PubMed

    Komthong, P; Suriyaphan, O; Charoenpanich, J

    2012-01-01

    Acrylamide in Thai-conventional snacks was analysed by GC/MS with a linear response ranged of 5-50 µg and r² > 0.99. The limit of detection (s/n = 3) and limit of quantification (s/n = 10) were 4 and 15 µg kg⁻¹, respectively, and RSD < 2%. Acrylamide in 19 food samples ranged from <15 µg kg⁻¹ to 1.26 mg kg⁻¹ with highest concentrations in Kanom Jak. Moderate levels (150-500 µg kg⁻¹) were detected mostly in deep-fried products, especially sweet potato and taro crisps, Kanom Kai Hong, banana fritters, durian chips and spring rolls. Thai-conventional snacks possessed low concentrations (<150 µg kg⁻¹) including Khao Larm, Pa Tong Koo, sweet banana crisps and deep-fried Chinese wonton. Acrylamide was lowest (<15 µg kg⁻¹) in fish strips, rice crackers, Hoi Jor and fried fish balls. Dietary habits by 400 tourists indicate a daily intake of acrylamide <150 ng, well below a toxic dose. PMID:24779691

  10. Similar Structures to the E-to-H Helix Unit in the Globin-Like Fold are Found in Other Helical Folds

    PubMed Central

    Matsuoka, Masanari; Fujita, Aoi; Kawai, Yosuke; Kikuchi, Takeshi

    2014-01-01

    A protein in the globin-like fold contains six alpha-helices, A, B, E, F, G and H. Among them, the E-to-H helix unit (E, F, G and H helices) forms a compact structure. In this study, we searched similar structures to the E-to-H helix of leghomoglobin in the whole protein structure space using the Dali program. Several similar structures were found in other helical folds, such as KaiA/RbsU domain and Type III secretion system domain. These observations suggest that the E-to-H helix unit may be a common subunit in the whole protein 3D structure space. In addition, the common conserved hydrophobic residues were found among the similar structures to the E-to-H helix unit. Hydrophobic interactions between the conserved residues may stabilize the 3D structures of the unit. We also predicted the possible compact regions of the units using the average distance method. PMID:24970216

  11. Molecular mechanisms of membrane targeting antibiotics.

    PubMed

    Epand, Richard M; Walker, Chelsea; Epand, Raquel F; Magarvey, Nathan A

    2016-05-01

    The bacterial membrane provides a target for antimicrobial peptides. There are two groups of bacteria that have characteristically different surface membranes. One is the Gram-negative bacteria that have an outer membrane rich in lipopolysaccharide. Several antimicrobials have been found to inhibit the synthesis of this lipid, and it is expected that more will be developed. In addition, antimicrobial peptides can bind to the outer membrane of Gram-negative bacteria and block passage of solutes between the periplasm and the cell exterior, resulting in bacterial toxicity. In Gram-positive bacteria, the major bacterial lipid component, phosphatidylglycerol can be chemically modified by bacterial enzymes to convert the lipid from anionic to cationic or zwitterionic form. This process leads to increased levels of resistance of the bacteria against polycationic antimicrobial agents. Inhibitors of this enzyme would provide protection against the development of bacterial resistance. There are antimicrobial agents that directly target a component of bacterial cytoplasmic membranes that can act on both Gram-negative as well as Gram-positive bacteria. Many of these are cyclic peptides with a rigid binding site capable of binding a lipid component. This binding targets antimicrobial agents to bacteria, rather than being toxic to host cells. This article is part of a Special Issue entitled: Antimicrobial peptides edited by Karl Lohner and Kai Hilpert. PMID:26514603

  12. Colonial modernity and networks in the Japanese empire: the role of Gotō Shinpei.

    PubMed

    Low, Morris

    2010-01-01

    This paper examines how Gotō Shinpei (1857-1929) sought to develop imperial networks emanating out of Tokyo in the fields of public health, railways, and communications. These areas helped define colonial modernity in the Japanese empire. In public health, Gotō's friendship with the bacteriologist Kitasato Shibasaburō led to the establishment of an Institute of Infectious Diseases in Tokyo. Key scientists from the institute took up positions in colonial medical colleges, creating a public health network that serviced the empire. Much of the empire itself was linked by a network of railways. Gotō was the first president of the South Manchuria Railway company (SMR). Communication technologies, especially radio, helped to bring the empire closer. By 1925, the Tokyo Broadcasting Station had begun its public radio broadcasts. Broadcasting soon came under the umbrella of the new organization, the Nippon Hōsō Kyōkai (NHK). Gotō was NHK's first president. The empire would soon be linked by radio, and it was by radio that Emperor Hirohito announced to the nation in 1945 that the empire had been lost. PMID:20549877

  13. Mono- and bis-tolyl­terpyridine iridium(III) complexes

    PubMed Central

    Hinkle, Lindsay M.; Young, Victor G.; Mann, Kent R.

    2010-01-01

    The first structure report of trichlorido[4′-(p-tolyl)-2,2′:6′,2′′-terpyridine]iridium(III) dimethyl sulfoxide solvate, [IrCl3(C22H17N3)]·C2H6OS, (I), is presented, along with a higher-symmetry setting of previously reported bis­[4′-(p-tolyl)-2,2′:6′,2′′-terpyridine]iridium(III) tris­(hexa­fluorido­phosphate) acetonitrile disolvate, [Ir(C22H17N3)2](PF6)3·2C2H3N, (II) [Yoshikawa, Yamabe, Kanehisa, Kai, Takashima & Tsukahara (2007 ▶). Eur. J. Inorg. Chem. pp. 1911–1919]. For (I), the data were collected with synchrotron radiation and the dimethyl sulfoxide solvent mol­ecule is disordered over three positions, one of which is an inversion center. The previously reported structure of (II) is presented in the more appropriate C2/c space group. The iridium complex and one PF6 − anion lie on twofold axes in this structure, making half of the mol­ecule unique. PMID:20203396

  14. Battlefield triage life signs detection techniques

    NASA Astrophysics Data System (ADS)

    Boric-Lubecke, Olga; Lin, Jenshan; Park, Byung-Kwon; Li, Changzhi; Massagram, Wansuree; Lubecke, Victor M.; Host-Madsen, Anders

    2008-04-01

    Getting to wounded soldiers on the battlefield is a precarious task, and medics have a very high casualty rate. It is therefore a vital importance to prioritize which soldiers to attend to first. The first step is to detect life signs - if a soldier is dead or alive, and prioritize recovery of live soldiers. The second step is to obtain vital signs from live soldiers, and use this to prioritize which are in most urgent need of attention. Our team at Kai Sensors, University of Hawaii and University of Florida is developing Doppler radar heart sensing technology that provides the means to detect life signs, respiration and/or heart beat, at a distance, even for subjects lying motionless, e.g., unconscious subjects, wearing body armor, and hidden from direct view. Since this technology can deliver heart rate information with high accuracy, it may also enable the assessment of a subject's physiological and psychological state based on heart rate variability (HRV) analysis. Thus, the degree of a subject's injury may also be determined. The software and hardware developments and challenges for life signs detection and monitoring for battlefield triage will be discussed, including heart signal detection from all four sides of the human body, detection in the presence of body armor, and the feasibility of HRV parameter extraction.

  15. Influence of pressure and silane depletion on microcrystalline silicon material quality and solar cell performance

    SciTech Connect

    Bugnon, G.; Feltrin, A.; Meillaud, F.; Ballif, C.; Bailat, J.

    2009-03-15

    Hydrogenated microcrystalline silicon growth by very high frequency plasma-enhanced chemical vapor deposition is investigated in an industrial-type parallel plate R and D KAI reactor to study the influence of pressure and silane depletion on material quality. Single junction solar cells with intrinsic layers prepared at high pressures and in high silane depletion conditions exhibit remarkable improvements, reaching 8.2% efficiency. Further analyses show that better cell performances are linked to a significant reduction of the bulk defect density in intrinsic layers. These results can be partly attributed to lower ion bombardment energies due to higher pressures and silane depletion conditions, improving the microcrystalline material quality. Layer amorphization with increasing power density is observed at low pressure and in low silane depletion conditions. A simple model for the average ion energy shows that ion energy estimates are consistent with the amorphization process observed experimentally. Finally, the material quality of a novel regime for high rate deposition is reviewed on the basis of these findings.

  16. Single mutations in sasA enable a simpler ΔcikA gene network architecture with equivalent circadian properties

    PubMed Central

    Shultzaberger, Ryan K.; Boyd, Joseph S.; Katsuki, Takeo; Golden, Susan S.; Greenspan, Ralph J.

    2014-01-01

    The circadian input kinase of the cyanobacterium Synechococcus elongatus PCC 7942 (CikA) is important both for synchronizing circadian rhythms with external environmental cycles and for transferring temporal information between the oscillator and the global transcriptional regulator RpaA (regulator of phycobilisome-associated A). KOs of cikA result in one of the most severely altered but still rhythmic circadian phenotypes observed. We chemically mutagenized a cikA-null S. elongatus strain and screened for second-site suppressor mutations that could restore normal circadian rhythms. We identified two independent mutations in the Synechococcus adaptive sensor A (sasA) gene that produce nearly WT rhythms of gene expression, likely because they compensate for the loss of CikA on the temporal phosphorylation of RpaA. Additionally, these mutations restore the ability to reset the clock after a short dark pulse through an output-independent pathway, suggesting that SasA can influence entrainment through direct interactions with KaiC, a property previously unattributed to it. These experiments question the evolutionary advantage of integrating CikA into the cyanobacterial clock, challenge the conventional construct of separable input and output pathways, and show how easily the cell can adapt to restore phenotype in a severely compromised genetic network. PMID:25385627

  17. The poor have become rich, and the rich have become poor: collective trauma in the Guinean Languette.

    PubMed

    Abramowitz, Sharon A

    2005-11-01

    This paper uses Kai Erikson's (Everything in its path: the disaster at Buffalo Creek. Simon & Schuster, New York, 1978) definition of collective trauma to interrogate the symptom reports and narrative accounts of six Guinean communities attacked by Sierra Leonean and Liberian RUF forces in 2000-2001. These data, collected in 2003, found high rates of fear, physical anxiety, emotional anxiety, depression, physical distress, sadness, and post-traumatic stress disorder-related symptoms across all communities, but found lower rates of distress among communities that had developed collective narratives of resistance to violence, or had concertedly resisted post-conflict social change. Communities with higher rates of distress tended to report community narratives of violence and post-conflict social life, which emphasized abandonment, isolation, disregard of community rituals and social supports, and the dislocation of local moral worlds. This study argues that the physical and emotional symptoms of trauma-related mental illness are articulations of collective trauma and represent the physical and emotional manifestations of the destruction of local moral worlds. It illuminates the processes by which violence inverts social experience, and argues that the social dimensions of trauma have long-term consequences for post-conflict reconstruction. PMID:16125293

  18. Nuclear waste`s human dimension

    SciTech Connect

    Erikson, K.; Colglazier, E.W.; White, G.F.

    1994-12-31

    The United States has pinned its hopes for a permanent underground repository for its high-level nuclear wastes on Yucca Mountain, Nevada. Nevertheless, the Department of Energy`s (DOE) site research efforts have failed {open_quotes}to adequately consider human behavior and emotions,{close_quotes} write Kai Erikson of Yale University, E. William Colglazier of the National Academy of Sciences, and Gilbert F. White of the University of Colorado. The authors maintain that it is impossible to predict changes in geology, seismology, and hydrology that may affect the Yucca Mountain area over the next 1,000 years. Predicting human behavior in that time frame remains even more daunting, they insist. They admit that {open_quotes}DOE...has been given the impossible assignment to take tens of thousands of metric tons of the most hazardous materials ever created and, in the face of growing opposition, entomb them so that they will do little harm for thousands of years.{close_quotes} The researchers suggest that the government seek a secure, retrievable storage arrangement while it continues its search for safer long-term options.

  19. Quantifying the robustness of circadian oscillations at the single-cell level

    NASA Astrophysics Data System (ADS)

    Lambert, Guillaume; Rust, Michael

    2014-03-01

    Cyanobacteria are light-harvesting microorganisms that contribute to 30% of the photosynthetic activity on Earth and contain one of the simplest circadian systems in the animal kingdom. In Synechococcus elongatus , a species of freshwater cyanobacterium, circadian oscillations are regulated by the KaiABC system, a trio of interacting proteins that act as a biomolecular pacemaker of the circadian system. While the core oscillator precisely anticipates Earth's 24h light/dark cycle, it is unclear how much individual cells benefit from the expression and maintenance of a circadian clock. By studying the growth dynamics of individual S . elongatus cells under sudden light variations, we show that several aspects of cellular growth, such as a cell's division probability and its elongation rate, are tightly coupled to the circadian clock. We propose that the evolution and maintenance of a circadian clock increases the fitness of cells by allowing them to take advantage of cyclical light/dark environments by alternating between two phenotypes: expansionary, where cells grow and divide at a fast pace during the first part of the day, and conservative, where cells enter a more quiescent state to better prepare to the stresses associated with the night's prolonged darkness.

  20. Application of preparative high-speed counter-current chromatography for separation and purification of arctiin from Fructus Arctii.

    PubMed

    Wang, Xiao; Li, Fuwei; Sun, Qinglei; Yuan, Jingpeng; Jiang, Ting; Zheng, Chengchao

    2005-01-21

    Following an initial clean-up step on the AB-8 resin (polystyrene resin, 0.3-1.25 mm: NanKai Chemical Factory, Tianjin, China), high-speed counter-current chromatography (HSCCC) was used to purify an arctiin from an extract of the fruits of the Arctium lappa L. Arctiin is a major lignan compound in the traditional Chinese medicinal herb A. lappa L. The two-phase solvent system used was composed of ethyl acetate-n-butanol-ethanol-water at an optimized volume ratio of 5:0.5:1:5 (v/v/v/v). The upper phase was used as the mobile phase in the head to tail elution mode. A total amount of 159 mg of arctiin at 98% purity was obtained from 350 mg of the crude extract (containing 49% arctiin) with 91% recovery. The preparative isolation and purification of arctiin by HSCCC was completed in 5 h in a separation. Identification of the target compound was performed by LC-electrospray ionization MS and 13C-NMR. The structure of the product was further confirmed by comparison with authentic sample (National Institute of the Control of Pharmaceutical and Biological Products, Beijing, China). PMID:15700478

  1. Downregulated long non-coding RNA MEG3 in breast cancer regulates proliferation, migration and invasion by depending on p53's transcriptional activity.

    PubMed

    Sun, Lin; Li, Yu; Yang, Bangxiang

    2016-09-01

    Long non-coding RNAs (lncRNAs) was found to play critical roles in tumorigenesis, hence, screen of tumor-related lncRNAs, identification of their biological roles is important for understanding the processes of tumorigenesis. In this study, we identified the expressing difference of several tumor-related lncRNAs in breast cancer samples and found that, MEG3, which is downregulated in non-small cell lung cancer (NSCLC) tumor tissues, is also downregulated in breast cancer samples compared with adjacent tissues. For figuring out the effect of MEG3 in breast cancer cells MCF7 and MB231, we overexpressed MEG3 in these cells, and found that it resulted the inhibition of proliferation, colony formation, migration and invasion capacities by enhancing p53's transcriptional activity on its target genes, including p21, Maspin and KAI1. MEG3 presented similar effects in MB157, which is a p53-null breast cancer cell line, when functional p53 but not p53R273H mutant, which lacks transcriptional activity, was introduced. Surprisingly, overexpression of MEG3 activates p53's transcriptional activity by decreasing MDM2's transcription level, and thus stabilizes and accumulates P53. Taken together, our findings indicate that MEG3 is downregulated in breast cancer tissues and affects breast cancer cells' malignant behaviors, which indicate MEG3 a potential therapeutic target for breast cancer. PMID:27166155

  2. Switching dynamics in ferroelectric P(VDF-TrFE) thin films

    NASA Astrophysics Data System (ADS)

    Zhao, Dong; Katsouras, Ilias; Asadi, Kamal; Blom, Paul W. M.; de Leeuw, Dago M.

    2015-12-01

    Switching, i.e., polarization reversal, of ferroelectric P(VDF-TrFE) thin films has been investigated in a wide range of applied electric field and temperature. The measured polarization transients can be quantitatively described by a compressed exponential function as originally formulated by Kolmogorov, Avrami, and Ishibashi (the KAI model). The phenomenological parameters switching time and Avrami index are related to the velocity and morphology of the domain walls, respectively. We show that the switching time depends exponentially on the electric field as described by the Merz law. The experimentally obtained Avrami index is independent of temperature but decreases with applied electric field from 1.55 at low field to 1.0 at 300 MV/m, indicative of an out-of-equilibrium dynamics of the growing domains. Using a random walk model we demonstrate that the observed switching dynamics is in agreement with a domain growth mechanism where domains are circular at low electric field with few nucleation sites, whereas at high field irregular entangled domains are formed. The density of nucleation sites is extracted as 1.5 % of the monomeric units in this work. Switching transients on capacitors where the nuclei density has been artificially increased exhibit one-dimensional-like growth, as predicted by our model.

  3. Reversible and Irreversible Magnetization Switching in Co/Pt Multilayers

    NASA Astrophysics Data System (ADS)

    Davies, J. E.; Liu, Kai; Hellwig, O.; Fullerton, E. E.

    2004-03-01

    The magnetization reversal processes in [Co(4ÅPt(7ÅN multilayers (N=5-150) with perpendicular anisotropy^1 have been investigated by a first order reversal curve (FORC) technique^2. In most samples, the resultant FORC distributions have three distinct regions: as the reversal magnetic field increases, the magnetization first goes through an irreversible switching process, the onset of which corresponds to the nucleation of reverse domains; interestingly a mostly reversible process is then observed, indicating the propagation of domains; finally a second irreversible process brings the sample to negative saturation, corresponding to the final annihilation of domains. For samples with larger numbers of repeats (N), the major loop becomes more and more slanted as the reverse domains form a labyrinth stripe domain state. Correspondingly, in the FORC distribution, the reversible region spans over a larger and larger field range. These irreversible and reversible processes are direct manifestations of reversal via domain nucleation and propagation, as well as rotation processes. ^1O. Hellwig, G. P. Denbeaux, J. B. Kortright, and E. E. Fullerton, Physica B 336, 136 (2003). ^2H. G. Katzgraber, F. Pázmándi, C. R. Pike, Kai Liu, R. T. Scalettar, K. L. Verosub, and G. T. Zimányi, Phys. Rev. Lett., 89, 257202 (2002).

  4. Influence of lipidation on the mode of action of a small RW-rich antimicrobial peptide.

    PubMed

    Wenzel, Michaela; Schriek, Patrick; Prochnow, Pascal; Albada, H Bauke; Metzler-Nolte, Nils; Bandow, Julia E

    2016-05-01

    Antimicrobial peptides are a potent class of antibiotics. In the Gram-positive model organism Bacillus subtilis the synthetic peptide RWRWRW-NH2 integrates into the bacterial membrane and delocalizes essential peripheral membrane proteins involved in cell wall biosynthesis and respiration. A lysine residue has been added to the hexapeptide core structure, either C or N-terminally. Lipidation of the lysine residues by a C8-acyl chain significantly improved antibacterial activity against both Gram-positive and Gram-negative bacteria. Here, we report a comparative proteomic study in B. subtilis on the mechanism of action of the lipidated and non-lipidated peptides. All derivatives depolarized the bacterial membrane without forming pores and all affected cell wall integrity. Proteomic profiling of the bacterial stress responses to the small RW-rich antimicrobial peptides was reflective of non-disruptive membrane integration. Overall, our results indicate that antimicrobial peptides can be derivatized with lipid chains enhancing antibacterial activity without significantly altering the mechanism of action. This article is part of a Special Issue entitled: Antimicrobial peptides edited by Karl Lohner and Kai Hilpert. PMID:26603779

  5. Cold Atmospheric Plasma for Medicine: State of Research and Clinical Application

    NASA Astrophysics Data System (ADS)

    von Woedtke, Thomas

    2015-09-01

    Basic research in plasma medicine has made excellent progress and resulted in the fundamental insights that biological effects of cold atmospheric plasmas (CAP) are significantly caused by changes of the liquid environment of cells, and are dominated by redox-active species. First CAP sources are CE-certified as medical devices. Main focus of plasma application is on wound healing and treatment of infective skin diseases. Clinical applications in this field confirm the supportive effect of cold plasma treatment in acceleration of healing of chronic wounds above all in cases where conventional treatment fails. Cancer treatment is another actual and emerging field of CAP application. The ability of CAP to kill cancer cells by induction of apoptosis has been proved in vitro. First clinical applications of CAP in palliative care of cancer are realized. In collaboration with Hans-Robert Metelmann, University Medicine Greifswald; Helmut Uhlemann, Klinikum Altenburger Land GmbH Altenburg; Anke Schmidt and Kai Masur, Leibniz Institute for Plasma Science and Technology (INP Greifswald); Renate Schönebeck, Neoplas Tools GmbH Greifswald; and Klaus-Dieter Weltmann, Leibniz Institute for Plasma Science and Technology (INP Greifswald).

  6. Spot centroid sensitivity to angle of intensity on image detector with lenslet array.

    PubMed

    Hui, Mei; Zhou, Ping; Su, Peng; Zhao, Zhu

    2015-05-20

    Lenslet array was introduced to an image detector to compensate for low sensitivity. These lenses deviate the light from different incident angles and potentially introduce errors when subpixel accuracy is needed. We investigated the spot centroid position because the angle of incidence changes on a Kodak KAI-16000 image detector with lenslet array. In our experiment, we noticed that there is a cubic dependency on the incident angle. The experimental results show that dependence on the angle of incidence is related to the lenslet array in the Kodak detector used for the pentaprism test. This situation caused an error in spherical aberration on the test surface after integration. The magnitude of the cubic component at incident angle of 14° (equivalent to F/2) is 11.6 μm, which corresponds to a 48 nm rms spherical aberration for the test surface and brings the scanning pentaprism test closer to the principal test while there is a 56 nm rms discrepancy. The discrepancy in spherical aberration between the two tests reduced to 8 nm after this calibration. It also showed the contrast measurement results for the Kodak detector and PointGrey detector. We performed experiments with two different detectors to quantify this effect. PMID:26192498

  7. Revealing the Atomic Site-Dependent g Factor within a Single Magnetic Molecule via the Extended Kondo Effect

    NASA Astrophysics Data System (ADS)

    Du, Shixuan

    Control over charge and spin states at the single molecule level is crucial not only for a fundamental understanding of charge and spin interactions but also represents a prerequisite for development of molecular electronics and spintronics. In this talk, I will talk about the extended spin distribution in space beyond the central Mn ion, and onto the non-magnetic constituent atoms of the MnPc molecule. This extended spin distribution results in an extended Kondo effect, which can be explained by spin polarization induced by symmetry breaking of the molecular framework, as confirmed by DFT calculations. Measuring the evolution of the Kondo splitting with applied magnetic fields at different atomic sites, we find a spatial variation of the g-factor within a single molecule for the first time. The existence of atomic site-dependent g-factors can be attributed to specific molecular orbitals distributed over the entire molecule. This work not only open up a new opportunity for quantum information recording, but also provide a new route to explore the internal electronic and spin structure of complex molecules, hard to achieve otherwise. (L. W. Liu et al., Phys. Rev. Lett. 2015, 114, 126601. In collaboration with Liwei Liu, Kai Yang, Yuhang Jiang, Li Gao, Qi Liu, Boqun Song, Wende Xiao, Haitao Zhou, Hongjun Gao in CAS, Min Ouyang in MU, and A.H. Castro Neto in SNU.) Revealing the Atomic Site-Dependent g Factor within a Single Magnetic Molecule via the Extended Kondo Effect.

  8. Interaction between antimicrobial peptides and mycobacteria.

    PubMed

    Gutsmann, Thomas

    2016-05-01

    Mycobacteria can cause different severe health problems, including tuberculosis (TB). The treatment of TB with conventional antibiotics is successful, however, the number of multi-drug and extensively-drug resistant Mycobacterium tuberculosis strains increases. Moreover, many classical antimycobacterial antibiotics have severe side effects. Therefore, antimicrobial peptides (AMPs) seem to be good candidates for new therapeutic strategies. On the one hand AMPs can be used as a single drug or in combination with conventional antibiotics to directly kill mycobacteria, or on the other hand to act as immunstimulatory agents. This review summarizes the findings on the role of endogenous human AMPs being involved in TB, the antimycobacterial activity of various AMPs, and the molecular modes of action. Most active AMPs interact with the mycobacterial cell envelope and in particular with the mycomembrane and the plasma membrane. The mycomembrane is a very rigid membrane probably leading to a lower activity of the AMPs against mycobacteria as compared to other Gram-negative or Gram-positive bacteria. For some AMPs also other targets have been identified. Because of the complex environment of intracellular mycobacteria being trapped in the phagosome, within the macrophage, within the granuloma, within the lung, the external administration of AMPs in the latent phase of TB is a challenge. However, in the acute phase the AMPs can attack mycobacteria in a direct way. This article is part of a Special Issue entitled: Antimicrobial peptides edited by Karl Lohner and Kai Hilpert. PMID:26851776

  9. Antimicrobial peptides and their interaction with biofilms of medically relevant bacteria.

    PubMed

    Batoni, Giovanna; Maisetta, Giuseppantonio; Esin, Semih

    2016-05-01

    Biofilm-associated infections represent one of the major threats of modern medicine. Biofilm-forming bacteria are encased in a complex mixture of extracellular polymeric substances (EPS) and acquire properties that render them highly tolerant to conventional antibiotics and host immune response. Therefore, there is a pressing demand of new drugs active against microbial biofilms. In this regard, antimicrobial peptides (AMPs) represent an option taken increasingly in consideration. After dissecting the peculiar biofilm features that may greatly affect the development of new antibiofilm drugs, the present article provides a general overview of the rationale behind the use of AMPs against biofilms of medically relevant bacteria and on the possible mechanisms of AMP-antibiofilm activity. An analysis of the interactions of AMPs with biofilm components, especially those constituting the EPS, and the obstacles and/or opportunities that may arise from such interactions in the development of new AMP-based antibiofilm strategies is also presented and discussed. This article is part of a Special Issue entitled: Antimicrobial Peptides edited by Karl Lohner and Kai Hilpert. PMID:26525663

  10. Determination of Vitamin C, b-carotene and Riboflavin Contents in Five Green Vegetables Organically and Conventionally Grown.

    PubMed

    Ismail, Amin; Cheah, Sook Fun

    2003-03-01

    As consumer interest in organically grown vegetables is increasing in Malaysia, there is a need to answer whether the vegetables are more nutritious than those conventionally grown. This study investigates commercially available vegetables grown organically and conventionally, purchased from retailers to analyse β-carotene, vitamin C and riboflavin contents. Five types of green vegetables were selected, namely Chinese mustard (sawi) (Brassica juncea), Chinese kale (kai-lan) (Brassica alboglabra), lettuce (daun salad) (Lactuca sativa), spinach (bayam putih) (Amaranthus viridis) and swamp cabbage (kangkung) (Ipomoea aquatica). For vitamin analysis, a reverse-phase high performance liquid chromatography was used to identify and quantify β -carotene, vitamin C and riboflavin. The findings showed that not all of the organically grown vegetables were higher in vitamins than that conventionally grown. This study found that only swamp cabbage grown organically was highest in β -carotene, vitamin C and riboflavin contents among the entire samples studied. The various nutrients in organically grown vegetables need to be analysed for the generation of a database on nutritional value which is important for future research. PMID:22692530

  11. Culture heritage and identity - some cases in Taiwan on the protection of cultural heritage

    NASA Astrophysics Data System (ADS)

    Wang, R. W.-C.

    2015-09-01

    The protection of cultural heritage relates to an issue of identity. How a nation or a state tries to face to its history is often revealed on the protection of cultural heritage. Taiwan is as a country with complex history, especially the period after World War II. This article will work on some significant cases, regarded as ideological representation of identity. This article works on the cultural identity by observing and analyzing different cases of classified Historic Monuments. In different political periods, we see how the government tries to fabricate on the identity issue by working on Historic Monuments preservation. During the presidency of Chiang Kai-shek and his son Chiang Ching-kuo, the classification of Historic Monuments tried to focus on those make by former Chinese migrants. They tried hard to establish and reaffirm the ever existing "fact" of people in Taiwan. Whereas after the late 1980s and 1990s, after Chiang's reign, local conscience has been awaken. Political ambience turned to a new era. This freedom of speech of post-Chiang's reign encourages people to seek on their identity. The complex political situation of Taiwan makes this seeking cultural identity related to the seeking of independence of Taiwan. The respect to the aboriginal people also reoriented to include the preservation of their tribes and villages.

  12. Global Transcriptomic Analysis Reveals the Mechanism of Phelipanche aegyptiaca Seed Germination

    PubMed Central

    Yao, Zhaoqun; Tian, Fang; Cao, Xiaolei; Xu, Ying; Chen, Meixiu; Xiang, Benchun; Zhao, Sifeng

    2016-01-01

    Phelipanche aegyptiaca is one of the most destructive root parasitic plants of Orobanchaceae. This plant has significant impacts on crop yields worldwide. Conditioned and host root stimulants, in particular, strigolactones, are needed for unique seed germination. However, no extensive study on this phenomenon has been conducted because of insufficient genomic information. Deep RNA sequencing, including de novo assembly and functional annotation was performed on P. aegyptiaca germinating seeds. The assembled transcriptome was used to analyze transcriptional dynamics during seed germination. Key gene categories involved were identified. A total of 274,964 transcripts were determined, and 53,921 unigenes were annotated according to the NR, GO, COG, KOG, and KEGG databases. Overall, 5324 differentially expressed genes among dormant, conditioned, and GR24-treated seeds were identified. GO and KEGG enrichment analyses demonstrated numerous DEGs related to DNA, RNA, and protein repair and biosynthesis, as well as carbohydrate and energy metabolism. Moreover, ABA and ethylene were found to play important roles in this process. GR24 application resulted in dramatic changes in ABA and ethylene-associated genes. Fluridone, a carotenoid biosynthesis inhibitor, alone could induce P. aegyptiaca seed germination. In addition, conditioning was probably not the indispensable stage for P. aegyptiaca, because the transcript level variation of MAX2 and KAI2 genes (relate to strigolactone signaling) was not up-regulated by conditioning treatment. PMID:27428962

  13. [Clinical usage of Yi-gan san-schizophrenia, borderline personality disorder, dyskinesia etc-].

    PubMed

    Horiguchi, Jyun

    2012-01-01

    Yi-gan san (YGS, yokukan-san in Japanese) was developed in 1555 by Xue Kai as a remedy for restlessness and agitation in children. Prompted by the increasing life expectancy of the Japanese population, geriatricians have begun to use this traditional regimen for behavioral and psychological symptoms of dementia in the elderly. Moreover, we reported that YGS therapy is a well-tolerated and effective remedy that improves the symptoms of schizophrenia., borderline personality disorder, Charles Bonnet Syndrome, pervasive developmental disorder. Asperger's disorder, neuroleptics induced tardive dyskinesia, and restless legs syndrome. In a pilot investigation, we administered YGS as an open-label adjunct to antipsychotic medication to patients with treatment-resistant shizophrenia, borderline personality disorder, Charles Bonnet Syndrome, pervasive developmental disorder, Asperger's disorder, neuroleptics induced tardive dyskinesia. The present lecture summarizes the available data based on the above our data. In addition, we extend our discussion to the potential applications of YGS for combining this treatment with cellular and molecular therapy. PMID:23094294

  14. Improving short antimicrobial peptides despite elusive rules for activity.

    PubMed

    Mikut, Ralf; Ruden, Serge; Reischl, Markus; Breitling, Frank; Volkmer, Rudolf; Hilpert, Kai

    2016-05-01

    Antimicrobial peptides (AMPs) can effectively kill a broad range of life threatening multidrug-resistant bacteria, a serious threat to public health worldwide. However, despite great hopes novel drugs based on AMPs are still rare. To accelerate drug development we studied different approaches to improve the antibacterial activity of short antimicrobial peptides. Short antimicrobial peptides seem to be ideal drug candidates since they can be synthesized quickly and easily, modified and optimized. In addition, manufacturing a short peptide drug will be more cost efficient than long and structured ones. In contrast to longer and structured peptides short AMPs seem hard to design and predict. Here, we designed, synthesized and screened five different peptide libraries, each consisting of 600 9-mer peptides, against Pseudomonas aeruginosa. Each library is presenting a different approach to investigate effectiveness of an optimization strategy. The data for the 3000 peptides were analyzed using models based on fuzzy logic bioinformatics and plausible descriptors. The rate of active or superior active peptides was improved from 31.0% in a semi-random library from a previous study to 97.8% in the best new designed library. This article is part of a Special Issue entitled: Antimicrobial peptides edited by Karl Lohner and Kai Hilpert. PMID:26687790

  15. Peptides with dual mode of action: Killing bacteria and preventing endotoxin-induced sepsis.

    PubMed

    Brandenburg, Klaus; Heinbockel, Lena; Correa, Wilmar; Lohner, Karl

    2016-05-01

    Bacterial infections, with the most severe form being sepsis, can often not be treated adequately leading to high morbidity and lethality of infected patients in critical care units. In particular, the increase in resistant bacterial strains and the lack of new antibiotics are main reasons for the worsening of the current situation, As a new approach, the use of antimicrobial peptides (AMPs) seems to be promising, combining the ability of broad-spectrum bactericidal activity and low potential of induction of resistance. Peptides based on natural defense proteins or polypeptides such as lactoferrin, Limulus anti-lipopolysaccharide factor (LALF), cathelicidins, and granulysins are candidates due to their high affinity to bacteria and to their pathogenicity factors, in first line lipopolysaccharide (LPS, endotoxin) of Gram-negative origin. In this review, we discuss literature with the focus on the use of AMPs from natural sources and their variants as antibacterial as well as anti-endotoxin (anti-inflammatory) drugs. Considerable progress has been made by the design of new AMPs for acting efficiently against the LPS-induced inflammation reaction in vitro as well as in vivo (mouse) models of sepsis. Furthermore, the data indicate that efficient antibacterial compounds are not necessarily equally efficient as anti-endotoxin drugs and vice versa. The most important reason for this may be the different molecular geometry of LPS in bacteria and in free form. This article is part of a Special Issue entitled: Antimicrobial peptides edited by Karl Lohner and Kai Hilpert. PMID:26801369

  16. On the controllers of prime ideals of group algebras of Abelian torsion-free groups of finite rank over a field of positive characteristic

    SciTech Connect

    Tushev, A V

    2006-10-31

    In the present paper certain methods are developed that enable one to study the properties of the controller of a prime faithful ideal I of the group algebra kA of an Abelian torsion-free group A of finite rank over a field k. The main idea is that the quotient ring kA/I by the given ideal I can be embedded as an integral domain k[A] into some field F and the group A becomes a subgroup of the multiplicative group of the field F. This allows one to apply certain results of field theory, such as Kummer's theory and the properties of the multiplicative groups of fields, to the study of the integral domain k[A]. In turn, the properties of the integral domain k[A]{approx_equal}kA/I depend essentially on the properties of the ideal I. In particular, by using these methods, an independent proof of the new version of Brookes's theorem on the controllers of prime ideals of the group algebra kA of an Abelian torsion-free group A of finite rank is obtained in the case where the field k has positive characteristic.

  17. Importance of active tectonics during karst formation. A Middle Eocene to Pleistocene example of the Lina Moutains (Irian Jaya, Indonesia)

    NASA Astrophysics Data System (ADS)

    Thery, J.-M.; Pubellier, M.; Thery, B.; Butterlin, J.; Blondeau, A.; Adams, C. G.

    1999-05-01

    The Lina Moutains show a typical example of karst formation associated to recent and active tectonics. The limestone samples were collected from giant potholes present beneath the heavy rainforest, during speleological expeditions to the Bird's Head of Irian-Jaya. Micropalaeontological data allow us to give a Middle Pleistocene age for the most recent karst formation. A detailed stratigraphy between the Upper Lutetian and the Middle Pleistocene was recorded, with tectonic events during the Oligocene and Pleistocene. The edge of the resurgence layer was also dated. We also conclude the probable existence of a subterraneous network downhill of the karst within the most recent levels of the Kais Limestone formation. We replace this formation within the tectonic evolution of this area between the Eocene and the Middle Pleistocene, in conjunction with the oblique convergence of the Pacific plate carrying volcanic arc fragments and the Australian margin, which resulted in folding, normal faulting associated with local extension, and wrench motion, which are settings capable of creating uplift of the carbonated platform.

  18. Mono- and bis-tolylterpyridine iridium(III) complexes

    SciTech Connect

    Hinkle, Lindsay M.; Young, Jr., Victor G.; Mann, Kent R.

    2012-01-20

    The first structure report of trichlorido[4'-(p-tolyl)-2,2':6',2{double_prime}-terpyridine]iridium(III) dimethyl sulfoxide solvate, [IrCl{sub 3}(C{sub 22}H{sub 17}N{sub 3})] {center_dot} C{sub 2}H{sub 6}OS, (I), is presented, along with a higher-symmetry setting of previously reported bis[4'-(p-tolyl)-2,2':6',2{double_prime}-terpyridine]iridium(III) tris(hexafluoridophosphate) acetonitrile disolvate, [Ir(C{sub 22}H{sub 17}N{sub 3})2](PF{sub 6}){sub 3} {center_dot} 2C{sub 2}H{sub 3}N, (II) [Yoshikawa, Yamabe, Kanehisa, Kai, Takashima & Tsukahara (2007). Eur. J. Inorg. Chem. pp. 1911-1919]. For (I), the data were collected with synchrotron radiation and the dimethyl sulfoxide solvent molecule is disordered over three positions, one of which is an inversion center. The previously reported structure of (II) is presented in the more appropriate C2/c space group. The iridium complex and one PF{sub 6}{sup -} anion lie on twofold axes in this structure, making half of the molecule unique.

  19. Single mutations in sasA enable a simpler ΔcikA gene network architecture with equivalent circadian properties.

    PubMed

    Shultzaberger, Ryan K; Boyd, Joseph S; Katsuki, Takeo; Golden, Susan S; Greenspan, Ralph J

    2014-11-25

    The circadian input kinase of the cyanobacterium Synechococcus elongatus PCC 7942 (CikA) is important both for synchronizing circadian rhythms with external environmental cycles and for transferring temporal information between the oscillator and the global transcriptional regulator RpaA (regulator of phycobilisome-associated A). KOs of cikA result in one of the most severely altered but still rhythmic circadian phenotypes observed. We chemically mutagenized a cikA-null S. elongatus strain and screened for second-site suppressor mutations that could restore normal circadian rhythms. We identified two independent mutations in the Synechococcus adaptive sensor A (sasA) gene that produce nearly WT rhythms of gene expression, likely because they compensate for the loss of CikA on the temporal phosphorylation of RpaA. Additionally, these mutations restore the ability to reset the clock after a short dark pulse through an output-independent pathway, suggesting that SasA can influence entrainment through direct interactions with KaiC, a property previously unattributed to it. These experiments question the evolutionary advantage of integrating CikA into the cyanobacterial clock, challenge the conventional construct of separable input and output pathways, and show how easily the cell can adapt to restore phenotype in a severely compromised genetic network. PMID:25385627

  20. The positive effects of typhoons on the fish CPUE in the South China Sea

    NASA Astrophysics Data System (ADS)

    Yu, Jie; Tang, Danling; Chen, Guobao; Li, Yongzhen; Huang, Zirong; Wang, Sufen

    2014-08-01

    Due to the logistical difficulties associated with fish data sampling after typhoons, short-term effects of typhoons on fishery in the South China Sea (SCS) have not been well-understood. The present study is to evaluate the impacts on the fish catch per unit effort (CPUE) owing to the three typhoons Chanthu, Vicente, and Kai-tak in the northwestern SCS, using long-term fish catch data and satellite data. The results show that the CPUE of total catch and some sorted catches have been changing because of the typhoons. On total catch, firstly, the CPUE has increased approximately 0.32 kg h-1 kw-1, 0.20 kg h-1 kw-1, and 0.25 kg h-1 kw-1 during the three typhoon periods. Then, the CPUEs decreased to the pre-typhoon level in about three weeks. Thirdly, among the three typhoons, the slow-moving Chanthu has caused a larger increase in CPUE. The typhoons impact was two-pronged, depending on fish species. One is the positive effects on meso-demersal fishes, cephalopoda and pelagic fishes. The other is the increase in CPUE of low trophic level carnivorous fishes after the three typhoons. This research provided the first evidence of CPUE increase after typhoons in the open sea.

  1. [The hospital ship Jutlandia].

    PubMed

    Winge, M

    1996-01-01

    The Danish contribution to the United Nations action during the Korean War (1950-52) was the hospital ship "Jutlandia". The motorvessel Jutlandia - 8.500 tons - was built by the Nakskov Shipyard in 1934, and was rebuilt in three months at the same shipyard to a modern hospital ship with 300 beds, 3 operating theatres, a dental clinic, an x-ray department etc. The crew and the hospital staff consisted approximately each of 100 persons. Jutlandia sailed for Korea on Jan. 23. 1951 and the expedition ended in Copenhagen on Oct. 16. 1953. On the first two cruises the ship was stationed at Pusan. During the first period mostly as an "evacuation sick-bay" and during the second period the ship was opened for Korean military and civil patients, and extensive help was given to the local population on shore. While in Denmark between the second and third cruise a helicopter deck was installed and the operating theatre for neuro-surgery was changed to an opthalmic clinic. This time the ship was stationed at the Bay of Ichon so close to the front, that the wounded could be admitted directly from the advanced dressing stations. On the return journeys to Europe patients were sailed to their home countries. Commodore Kai Hammerich was in charge of the expedition and captain Christen Kondrup was in charge of the ship, throughout the whole expedition. PMID:11625136

  2. Areal Feature Matching Based on Similarity Using Critic Method

    NASA Astrophysics Data System (ADS)

    Kim, J.; Yu, K.

    2015-10-01

    In this paper, we propose an areal feature matching method that can be applied for many-to-many matching, which involves matching a simple entity with an aggregate of several polygons or two aggregates of several polygons with fewer user intervention. To this end, an affine transformation is applied to two datasets by using polygon pairs for which the building name is the same. Then, two datasets are overlaid with intersected polygon pairs that are selected as candidate matching pairs. If many polygons intersect at this time, we calculate the inclusion function between such polygons. When the value is more than 0.4, many of the polygons are aggregated as single polygons by using a convex hull. Finally, the shape similarity is calculated between the candidate pairs according to the linear sum of the weights computed in CRITIC method and the position similarity, shape ratio similarity, and overlap similarity. The candidate pairs for which the value of the shape similarity is more than 0.7 are determined as matching pairs. We applied the method to two geospatial datasets: the digital topographic map and the KAIS map in South Korea. As a result, the visual evaluation showed two polygons that had been well detected by using the proposed method. The statistical evaluation indicates that the proposed method is accurate when using our test dataset with a high F-measure of 0.91.

  3. An intimate link between antimicrobial peptide sequence diversity and binding to essential components of bacterial membranes.

    PubMed

    Schmitt, Paulina; Rosa, Rafael D; Destoumieux-Garzón, Delphine

    2016-05-01

    Antimicrobial peptides and proteins (AMPs) are widespread in the living kingdom. They are key effectors of defense reactions and mediators of competitions between organisms. They are often cationic and amphiphilic, which favors their interactions with the anionic membranes of microorganisms. Several AMP families do not directly alter membrane integrity but rather target conserved components of the bacterial membranes in a process that provides them with potent and specific antimicrobial activities. Thus, lipopolysaccharides (LPS), lipoteichoic acids (LTA) and the peptidoglycan precursor Lipid II are targeted by a broad series of AMPs. Studying the functional diversity of immune effectors tells us about the essential residues involved in AMP mechanism of action. Marine invertebrates have been found to produce a remarkable diversity of AMPs. Molluscan defensins and crustacean anti-LPS factors (ALF) are diverse in terms of amino acid sequence and show contrasted phenotypes in terms of antimicrobial activity. Their activity is directed essentially against Gram-positive or Gram-negative bacteria due to their specific interactions with Lipid II or Lipid A, respectively. Through those interesting examples, we discuss here how sequence diversity generated throughout evolution informs us on residues required for essential molecular interaction at the bacterial membranes and subsequent antibacterial activity. Through the analysis of molecular variants having lost antibacterial activity or shaped novel functions, we also discuss the molecular bases of functional divergence in AMPs. This article is part of a Special Issue entitled: Antimicrobial peptides edited by Karl Lohner and Kai Hilpert. PMID:26498397

  4. Electromagnetic mixed-waste processing system for asbestos decontamination

    SciTech Connect

    1995-04-01

    The first phase of a program to develop and demonstrate a cost-effective, integrated process for remediation of asbestos-containing material that is contaminated with organics, heavy metals, and radioactive compounds was successfully completed. Laboratory scale tests were performed to demonstrate initial process viability for asbestos conversion, organics removal, and radionuclide and heavy metal removal. All success criteria for the laboratory tests were met. (1) Ohio DSI demonstrated greater than 99% asbestos conversion to amorphous solids using their commercial process. (2) KAI demonstrated 90% removal of organics from the asbestos suspension. (3) Westinghouse STC achieved the required metals removal criteria on a laboratory scale (e.g., 92% removal of uranium from solution, resin loadings of 0.6 equivalents per liter, and greater than 50% regeneration of resin in a batch test.) Using the information gained in the laboratory tests, the process was reconfigured to provide the basis for the mixed waste remediation system. An integrated process is conceptually developed, and a Phase 2 program plan is proposed to provide the bench-scale development needed in order to refine the design basis for a pilot processing system.

  5. Advancing Cancer Survivorship in a Country with 1.35 Billion People: The China Lymphoma Project

    PubMed Central

    Coughlin, Steven; Reno, Jamie

    2016-01-01

    Rates of lymphoma are rising rapidly and lymphoma is now the ninth most common cancer among Chinese males. The China Lymphoma Project was founded to increase awareness of lymphoma in China, including the survivability of the disease and the availability of potentially life-saving treatments, and to provide social support for men, women, and children in China who are living with the disease. The project is working with China government officials, several of the top cancer hospitals in China and the U.S., internationally known oncologists and cancer researchers, pharmaceutical and biotech companies in China and the U.S., healthcare and environmental companies, the Confucius Institute at San Diego State University, and the Asian Heritage Society. Advances in e-Health are being utilized to provide patient education and social support. The project will provide free e-books that profile lymphoma survivors (e.g., Kai-Fu Lee, creator of Google China), new videos, websites, pamphlets, blogs, video logs (vlogs), peer-to-peer counseling and support, and information about the latest treatments and oncology clinical trials.

  6. Co-localization of the amyloid precursor protein and the Notch intracellular domains in nuclear transcription factories

    PubMed Central

    Konietzko, Uwe; Goodger, Zoë V.; Meyer, Michelle; Kohli, Bernhard M.; Bosset, Jérôme; Lahiri, Debomoy K.; Nitsch, Roger M.

    2009-01-01

    The β-amyloid precursor protein (APP) plays a major role in Alzheimer’s disease. The APP intracellular domain (AICD), together with Fe65 and Tip60, localizes to spherical nuclear AFT complexes that might represent sites of transcription. We now show that endogenous AICD is targeted to similar nuclear spots. AFT complexes were closely associated with Cajal and PML bodies but did not localize to nucleoli or splicing speckles. Live imaging revealed that AFT complexes were highly mobile within nuclei. Following pharmacological inhibition of transcription AFT complexes merged into a few large assemblies. We have previously shown that AICD regulates the expression of its own precursor APP. Transfection of APP promoter plasmids as substrates resulted in cytosolic AFT complex formation at the labeled APP promoter plasmids. In addition, identification of chromosomal APP or KAI1 gene loci by fluorescence in situ hybridization showed their close association with nuclear AFT complexes. The transcriptional activator Notch intracellular domain (NICD) localized to the same nuclear spots as occupied by AFT complexes, suggesting that these nuclear compartments correspond to transcription factories. Fe65 and Tip60 also co-localized with APP in the neurites of primary neurons. Pre-assembled AFT complexes may serve to assist fast nuclear signaling upon endoproteolytic APP cleavage. PMID:18403052

  7. Regional release of [3H]adenosine derivatives from rat brain in vivo: effect of excitatory amino acids, opiate agonists, and benzodiazepines.

    PubMed

    Jhamandas, K; Dumbrille, A

    1980-11-01

    Previous studies have suggested that the theophylline-sensitive actions of (a) glutamic acid and related excitatory amino acids, (b) opiate agonists, and (c) benzodiazepines may be mediated by adenosine and its derivatives (AD). In this study the effects of these theophylline-sensitive agents on the in vivo release of labelled AD from the rat cerebral cortex and cerebellum were examined. Application of L-glutamate (L-Glu) to the cortex or cerebellum, preloaded with [3H]adenosine or [3H]adenine, markedly stimulated the release of [3H]adenosine derivatives ([3H]AD) from these regions. This effect was also produced by L-aspartate (L-Asp), kainate (Kai), and two "non-amino-acid" depolarizing agents, veratridine and potassium (56 mM). In the cerebral cortex the excitatory action of L-Glu or L-Asp was stronger than that of Kai but in the cerebellum all of these agents produced a similar effect on the release of [3H]AD. Pretreatment of the cortex with glutamic acid diethylester, HA966, 2-amino-4-phosphonobutyrate, and 2,3-diaminopropionate, but not theophylline, selectively antagonized the releasing action of L-Glu. This action of L-Glu was also reduced by EGTA added to a Ca-free physiological medium. In the cortex and cerebellum the excitatory action of L-Glu on the release of [3H]AD was attenuated by tetrodotoxin (TTX). TTX also antagonized the action of veratridine, but not potassium, on the cortical output of [3H]AD. Chromatographic analysis of release samples showed that about 60% of the total radioactivity released by L-Glu was associated with pharmacologically active AD, adenosine, and adenine nucleotides. The opiate agonists morphine and D-Met2-Pro5-enkephalinamide by themselves did not influence the spontaneous release of [3H]AD from the cortex but both agents very effectively inhibited the release evoked by L-Glu. This inhibitory action was antagonized by naloxone. At a low dose levorphanol, but not dextrorphan, inhibited the L-Glu-evoked release of [3H]AD. Both

  8. Education of Sustainability Engineers

    NASA Astrophysics Data System (ADS)

    Oleschko, K.; Perrier, E.; Tarquis, A. M.

    2010-05-01

    It's not the same to educate the sustainable engineers as to prepare the engineers of Sustainability. In the latter case all existing methods of inventive creativity (Altshuller, 1988) should be introduced in the teaching and research processes in order to create a culture of innovation at a group. The Theory of Inventing Problem Solving (TRIZ) is based on the pioneer works of Genrich Altshuller (1988) and his associated. Altshuller reviewed over 2 million patents beginning in 1946 (Orlov, 2006) and developed the Laws of Evolution of Technological Systems; An Algorithm for Inventive Problem Solving (ARIZ); forty typical Techniques for Overcoming System Conflicts (TOSC); a system of 76 Standard Approaches to Inventive Problems (Standards) etc. (Fey and Rivin, 1997). Nowadays, "a theory and constructive instrument package for the controlled synthesis of ideas and the focused transformation of the object to be improved" (Orlov, 2006) are used with high efficacy as the teaching and thinking inventive problem-solving methods in some high schools (Barak and Mesika, 2006; Sokoi et al., 2008) as well as a framework for research (Moehrle, 2005) in construction industry (Zhang et al., 2009); chemical engineering (Cortes Robles et al., 2008) etc. In 2005 US Congress passed the innovation act with the intent of increasing research investment (Gupta, 2007), while China had included inventive principles of TRIZ in strategy and decision making structure design (Kai Yang, 2010). The integrating of TRIZ into eco-innovation diminishes the common conflicts between technology and environment (Chang and Chen, 2004). In our presentation we show discuss some examples of future patents elaborated by the master degree students of Queretaro University, Faculty of Engineering, Mexico using TRIZ methods. References 1. Altshuller, G., 1988. Creativity as an Exact Science. Gordon and Breach, New York. 2. Chang, Hsiang-Tang and Chen, Jahau Lewis, 2004. The conflict-problem-solving CAD software

  9. The bioinformatics of nucleotide sequence coding for proteins requiring metal coenzymes and proteins embedded with metals

    NASA Astrophysics Data System (ADS)

    Tremberger, G.; Dehipawala, Sunil; Cheung, E.; Holden, T.; Sullivan, R.; Nguyen, A.; Lieberman, D.; Cheung, T.

    2015-09-01

    All metallo-proteins need post-translation metal incorporation. In fact, the isotope ratio of Fe, Cu, and Zn in physiology and oncology have emerged as an important tool. The nickel containing F430 is the prosthetic group of the enzyme methyl coenzyme M reductase which catalyzes the release of methane in the final step of methano-genesis, a prime energy metabolism candidate for life exploration space mission in the solar system. The 3.5 Gyr early life sulfite reductase as a life switch energy metabolism had Fe-Mo clusters. The nitrogenase for nitrogen fixation 3 billion years ago had Mo. The early life arsenite oxidase needed for anoxygenic photosynthesis energy metabolism 2.8 billion years ago had Mo and Fe. The selection pressure in metal incorporation inside a protein would be quantifiable in terms of the related nucleotide sequence complexity with fractal dimension and entropy values. Simulation model showed that the studied metal-required energy metabolism sequences had at least ten times more selection pressure relatively in comparison to the horizontal transferred sequences in Mealybug, guided by the outcome histogram of the correlation R-sq values. The metal energy metabolism sequence group was compared to the circadian clock KaiC sequence group using magnesium atomic level bond shifting mechanism in the protein, and the simulation model would suggest a much higher selection pressure for the energy life switch sequence group. The possibility of using Kepler 444 as an example of ancient life in Galaxy with the associated exoplanets has been proposed and is further discussed in this report. Examples of arsenic metal bonding shift probed by Synchrotron-based X-ray spectroscopy data and Zn controlled FOXP2 regulated pathways in human and chimp brain studied tissue samples are studied in relationship to the sequence bioinformatics. The analysis results suggest that relatively large metal bonding shift amount is associated with low probability correlation R

  10. Measuring the style of innovative thinking among engineering students

    NASA Astrophysics Data System (ADS)

    Passig, David; Cohen, Lizi

    2014-01-01

    Background: Many tools have been developed to measure the ability of workers to innovate. However, all of them are based on self-reporting questionnaires, which raises questions about their validity Purpose: The aim was to develop and validate a tool, called Ideas Generation Implementation (IGI), to objectively measure the style and potential of engineering students in generating innovative technological ideas. The cognitive framework of IGI is based on the Architectural Innovation Model (AIM). Tool description: The IGI tool was designed to measure the level of innovation in generating technological ideas and their potential to be implemented. These variables rely on the definition of innovation as 'creativity, implemented in a high degree of success'. The levels of innovative thinking are based on the AIM and consist of four levels: incremental innovation, modular innovation, architectural innovation and radical innovation. Sample: Sixty experts in technological innovation developed the tool. We checked its face validity and calculated its reliability in a pilot study (kappa = 0.73). Then, 145 undergraduate students were sampled at random from the seven Israeli universities offering engineering programs and asked to complete the questionnaire. Design and methods: We examined the construct validity of the tool by conducting a variance analysis and measuring the correlations between the innovator's style of each student, as suggested by the AIM, and the three subscale factors of creative styles (efficient, conformist and original), as suggested by the Kirton Adaptors and Innovators (KAI) questionnaire. Results: Students with a radical innovator's style inclined more than those with an incremental innovator's style towards the three creative cognitive styles. Students with an architectural innovator's style inclined moderately, but not significantly, towards the three creative styles. Conclusions: The IGI tool objectively measures innovative thinking among students