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Sample records for activities impart important

  1. Can exergames impart health messages? Game play, framing, and drivers of physical activity among children.

    PubMed

    Lwin, May O; Malik, Shelly

    2014-01-01

    This study examines the effectiveness of incorporating exergaming into physical education lessons as a platform for imparting health education messages and influencing children's beliefs about and attitudes toward physical activity. The authors launched a 6-week intervention program using Nintendo Wii games coupled with protection motivation theory-based health messaging among 5th-grade school children in Singapore. Results indicated that when children who were exposed to threat-framed messages played Wii exergames during physical education lessons, they reported more positive physical activity attitude, self-efficacy, and perceived behavioral control than did those who underwent regular physical education lessons and were exposed to the same message. In addition, among children playing Wii, the threat and coping frames had similar effects on the degree of message influence on physical activity attitudes and beliefs. The implications for schools, parents, and health policy are discussed.

  2. Can exergames impart health messages? Game play, framing, and drivers of physical activity among children.

    PubMed

    Lwin, May O; Malik, Shelly

    2014-01-01

    This study examines the effectiveness of incorporating exergaming into physical education lessons as a platform for imparting health education messages and influencing children's beliefs about and attitudes toward physical activity. The authors launched a 6-week intervention program using Nintendo Wii games coupled with protection motivation theory-based health messaging among 5th-grade school children in Singapore. Results indicated that when children who were exposed to threat-framed messages played Wii exergames during physical education lessons, they reported more positive physical activity attitude, self-efficacy, and perceived behavioral control than did those who underwent regular physical education lessons and were exposed to the same message. In addition, among children playing Wii, the threat and coping frames had similar effects on the degree of message influence on physical activity attitudes and beliefs. The implications for schools, parents, and health policy are discussed. PMID:24191779

  3. Taccalonolide binding to tubulin imparts microtubule stability and potent in vivo activity

    PubMed Central

    Risinger, AL; Li, J; Bennett, MJ; Rohena, CC; Peng, J; Schriemer, DC; Mooberry, SL

    2013-01-01

    The taccalonolides are highly acetylated steroids that stabilize cellular microtubules and overcome multiple mechanisms of taxane resistance. Recently, two potent taccalonolides, AF and AJ, were identified that bind tubulin directly and enhance microtubule polymerization. Extensive studies were conducted to characterize these new taccalonolides. AF and AJ caused aberrant mitotic spindles and bundling of interphase microtubules that differed from the effects of either paclitaxel or laulimalide. AJ also distinctly affected microtubule polymerization in that it enhanced the rate and extent of polymerization in the absence of any noticeable effect on microtubule nucleation. Additionally, the resulting microtubules were found to be profoundly cold stable. These data, along with studies showing synergistic antiproliferative effects between AJ and either paclitaxel or laulimalide, suggest a distinct binding site. Direct binding studies demonstrated that AJ could not be displaced from microtubules by paclitaxel, laulimalide or denaturing conditions, suggesting irreversible binding of AJ to microtubules. Mass spectrometry confirmed a covalent interaction of AJ with a peptide of β-tubulin containing the cyclostreptin binding sites. Importantly, AJ imparts strong inter-protofilament stability in a manner different from other microtubule stabilizers that covalently bind tubulin, consistent with the distinct effects of the taccalonolides as compared to other stabilizers. AF was found to be a potent and effective antitumor agent that caused tumor regression in the MDA-MB-231 breast cancer xenograft model. The antitumor efficacy of some taccalonolides, which stabilize microtubules in a manner different from other microtubule stabilizers, provides the impetus to explore the therapeutic potential of this site. PMID:24048820

  4. Modular activation of Rho1 by GPCR signalling imparts polarized myosin II activation during morphogenesis.

    PubMed

    Kerridge, Stephen; Munjal, Akankshi; Philippe, Jean-Marc; Jha, Ankita; de las Bayonas, Alain Garcia; Saurin, Andrew J; Lecuit, Thomas

    2016-03-01

    Polarized cell shape changes during tissue morphogenesis arise by controlling the subcellular distribution of myosin II. For instance, during Drosophila melanogaster gastrulation, apical constriction and cell intercalation are mediated by medial-apical myosin II pulses that power deformations, and polarized accumulation of myosin II that stabilizes these deformations. It remains unclear how tissue-specific factors control different patterns of myosin II activation and the ratchet-like myosin II dynamics. Here we report the function of a common pathway comprising the heterotrimeric G proteins Gα12/13, Gβ13F and Gγ1 in activating and polarizing myosin II during Drosophila gastrulation. Gα12/13 and the Gβ13F/γ1 complex constitute distinct signalling modules, which regulate myosin II dynamics medial-apically and/or junctionally in a tissue-dependent manner. We identify a ubiquitously expressed GPCR called Smog required for cell intercalation and apical constriction. Smog functions with other GPCRs to quantitatively control G proteins, resulting in stepwise activation of myosin II and irreversible cell shape changes. We propose that GPCR and G proteins constitute a general pathway for controlling actomyosin contractility in epithelia and that the activity of this pathway is polarized by tissue-specific regulators. PMID:26780298

  5. An ionizable active-site tryptophan imparts catalase activity to a peroxidase core.

    PubMed

    Loewen, Peter C; Carpena, Xavi; Vidossich, Pietro; Fita, Ignacio; Rovira, Carme

    2014-05-21

    Catalase peroxidases (KatG's) are bifunctional heme proteins that can disproportionate hydrogen peroxide (catalatic reaction) despite their structural dissimilarity with monofunctional catalases. Using X-ray crystallography and QM/MM calculations, we demonstrate that the catalatic reaction of KatG's involves deprotonation of the active-site Trp, which plays a role similar to that of the distal His in monofunctional catalases. The interaction of a nearby mobile arginine with the distal Met-Tyr-Trp essential adduct (in/out) acts as an electronic switch, triggering deprotonation of the adduct Trp.

  6. A peroxisome proliferator-activated receptor ligand MCC-555 imparts anti-proliferative response in pancreatic cancer cells by PPARgamma-independent up-regulation of KLF4

    SciTech Connect

    Min, Kyung-Won; Zhang, Xiaobo; Imchen, Temjenmongla; Baek, Seung Joon

    2012-09-01

    MCC-555 is a novel PPARα/γ dual ligand of the thiazolidinedione class and was recently developed as an anti-diabetic drug with unique properties. MCC-555 also has anti-proliferative activity through growth inhibition and apoptosis induction in several cancer cell types. Our group has shown that MCC-555 targets several proteins in colorectal tumorigenesis including nonsteroidal anti-inflammatory drug (NSAID)-activated gene (NAG-1) which plays an important role in chemoprevention responsible for chemopreventive compounds. NAG-1 is a member of the TGF-β superfamily and is involved in tumor progression and development; however, NAG-1's roles in pancreatic cancer have not been studied. In this report, we found that MCC-555 alters not only NAG-1 expression, but also p21 and cyclin D1 expression. NAG-1 and p21 expression was not blocked by PPARγ-specific antagonist GW9662, suggesting that MCC-555-induced NAG-1 and p21 expression is independent of PPARγ activation. However, decreasing cyclin D1 by MCC-555 seems to be affected by PPARγ activation. Further, we found that the GC box located in the NAG-1 promoter play an important role in NAG-1 transactivation by MCC-555. Subsequently, we screened several transcription factors that may bind to the GC box region in the NAG-1 promoter and found that KLF4 potentially binds to this region. Expression of KLF4 precedes NAG-1 and p21 expression in the presence of MCC-555, whereas blocking KLF4 expression using specific KLF4 siRNA showed that both NAG-1 and p21 expression by MCC-555 was blocked. In conclusion, MCC-555's actions on anti-proliferation involve both PPARγ-dependent and -independent pathways, thereby enhancing anti-tumorigenesis in pancreatic cancer cells. -- Highlights: ► PPARα/γ ligand MCC-555 exhibits anti-proliferative activity in pancreatic cancer cells. ► MCC-555 affects KLF4 expression following by NAG-1 and p21 expression in a PPARγ independent manner. ► MCC-555 also affects cyclin D1 down

  7. Calcium-activated and voltage-gated potassium channels of the pancreatic islet impart distinct and complementary roles during secretagogue induced electrical responses

    PubMed Central

    Jacobson, David A; Mendez, Felipe; Thompson, Michael; Torres, Jacqueline; Cochet, Olivia; Philipson, Louis H

    2010-01-01

    Glucose-induced β-cell action potential (AP) repolarization is regulated by potassium efflux through voltage gated (Kv) and calcium activated (KCa) potassium channels. Thus, ablation of the primary Kv channel of the β-cell, Kv2.1, causes increased AP duration. However, Kv2.1−/− islet electrical activity still remains sensitive to the potassium channel inhibitor tetraethylammonium. Therefore, we utilized Kv2.1−/− islets to characterize Kv and KCa channels and their respective roles in modulating the β-cell AP. The remaining Kv current present in Kv2.1−/−β-cells is inhibited with 5 μm CP 339818. Inhibition of the remaining Kv current in Kv2.1−/− mouse β-cells increased AP firing frequency by 39.6% but did not significantly enhance glucose stimulated insulin secretion (GSIS). The modest regulation of islet AP frequency by CP 339818 implicates other K+ channels, possibly KCa channels, in regulating AP repolarization. Blockade of the KCa channel BK with slotoxin increased β-cell AP amplitude by 28.2%, whereas activation of BK channels with isopimaric acid decreased β-cell AP amplitude by 30.6%. Interestingly, the KCa channel SK significantly contributes to Kv2.1−/− mouse islet AP repolarization. Inhibition of SK channels decreased AP firing frequency by 66% and increased AP duration by 67% only when Kv2.1 is ablated or inhibited and enhanced GSIS by 2.7-fold. Human islets also express SK3 channels and their β-cell AP frequency is significantly accelerated by 4.8-fold with apamin. These results uncover important repolarizing roles for both Kv and KCa channels and identify distinct roles for SK channel activity in regulating calcium- versus sodium-dependent AP firing. PMID:20643768

  8. Preparation of silver nano-particles immobilized onto chitin nano-crystals and their application to cellulose paper for imparting antimicrobial activity.

    PubMed

    Li, Zhihan; Zhang, Ming; Cheng, Dong; Yang, Rendang

    2016-10-20

    Immobilized silver nano-particles (Ag NPs) possess excellent antimicrobial properties due to their unique surface characteristics. In this paper, immobilized silver nano-particles were synthesized in the presence of chitin nano-crystals (CNC) based on the Tollens mechanism (reduction of silver ion by aldehydes in the chitosan oligosaccharides (COS)) under microwave-assisted conditions. The prepared Ag NPs-loaded CNC nano-composites were then applied onto the paper surface via coating for the preparation of antibacterial paper. Fourier transform infrared (FT-IR) and X-ray diffraction (XRD) results confirmed that the Ag NPs were immobilized onto the CNC. The transmission electron microscope (TEM) and scanning electron microscopy (SEM) results further revealed that the spherical Ag NPs (5-12nm) were well dispersed on the surface of CNC. The coated paper made from the Ag NPs-loaded CNC nano-composites exhibited a high effectiveness of the antibacterial activity against E. coli or S. aureus.

  9. Preparation of silver nano-particles immobilized onto chitin nano-crystals and their application to cellulose paper for imparting antimicrobial activity.

    PubMed

    Li, Zhihan; Zhang, Ming; Cheng, Dong; Yang, Rendang

    2016-10-20

    Immobilized silver nano-particles (Ag NPs) possess excellent antimicrobial properties due to their unique surface characteristics. In this paper, immobilized silver nano-particles were synthesized in the presence of chitin nano-crystals (CNC) based on the Tollens mechanism (reduction of silver ion by aldehydes in the chitosan oligosaccharides (COS)) under microwave-assisted conditions. The prepared Ag NPs-loaded CNC nano-composites were then applied onto the paper surface via coating for the preparation of antibacterial paper. Fourier transform infrared (FT-IR) and X-ray diffraction (XRD) results confirmed that the Ag NPs were immobilized onto the CNC. The transmission electron microscope (TEM) and scanning electron microscopy (SEM) results further revealed that the spherical Ag NPs (5-12nm) were well dispersed on the surface of CNC. The coated paper made from the Ag NPs-loaded CNC nano-composites exhibited a high effectiveness of the antibacterial activity against E. coli or S. aureus. PMID:27474631

  10. Imparting passivity to vapor deposited magnesium alloys

    NASA Astrophysics Data System (ADS)

    Wolfe, Ryan C.

    Magnesium has the lowest density of all structural metals. Utilization of low density materials is advantageous from a design standpoint, because lower weight translates into improved performance of engineered products (i.e., notebook computers are more portable, vehicles achieve better gas mileage, and aircraft can carry more payload). Despite their low density and high strength to weight ratio, however, the widespread implementation of magnesium alloys is currently hindered by their relatively poor corrosion resistance. The objective of this research dissertation is to develop a scientific basis for the creation of a corrosion resistant magnesium alloy. The corrosion resistance of magnesium alloys is affected by several interrelated factors. Among these are alloying, microstructure, impurities, galvanic corrosion effects, and service conditions, among others. Alloying and modification of the microstructure are primary approaches to controlling corrosion. Furthermore, nonequilibrium alloying of magnesium via physical vapor deposition allows for the formation of single-phase magnesium alloys with supersaturated concentrations of passivity-enhancing elements. The microstructure and surface morphology is also modifiable during physical vapor deposition through the variation of evaporation power, pressure, temperature, ion bombardment, and the source-to-substrate distance. Aluminum, titanium, yttrium, and zirconium were initially chosen as candidates likely to impart passivity on vapor deposited magnesium alloys. Prior to this research, alloys of this type have never before been produced, much less studied. All of these metals were observed to afford some degree of corrosion resistance to magnesium. Due to the especially promising results from nonequilibrium alloying of magnesium with yttrium and titanium, the ternary magnesium-yttrium-titanium system was investigated in depth. While all of the alloys are lustrous, surface morphology is observed under the scanning

  11. Imparting passivity to vapor deposited magnesium alloys

    NASA Astrophysics Data System (ADS)

    Wolfe, Ryan C.

    Magnesium has the lowest density of all structural metals. Utilization of low density materials is advantageous from a design standpoint, because lower weight translates into improved performance of engineered products (i.e., notebook computers are more portable, vehicles achieve better gas mileage, and aircraft can carry more payload). Despite their low density and high strength to weight ratio, however, the widespread implementation of magnesium alloys is currently hindered by their relatively poor corrosion resistance. The objective of this research dissertation is to develop a scientific basis for the creation of a corrosion resistant magnesium alloy. The corrosion resistance of magnesium alloys is affected by several interrelated factors. Among these are alloying, microstructure, impurities, galvanic corrosion effects, and service conditions, among others. Alloying and modification of the microstructure are primary approaches to controlling corrosion. Furthermore, nonequilibrium alloying of magnesium via physical vapor deposition allows for the formation of single-phase magnesium alloys with supersaturated concentrations of passivity-enhancing elements. The microstructure and surface morphology is also modifiable during physical vapor deposition through the variation of evaporation power, pressure, temperature, ion bombardment, and the source-to-substrate distance. Aluminum, titanium, yttrium, and zirconium were initially chosen as candidates likely to impart passivity on vapor deposited magnesium alloys. Prior to this research, alloys of this type have never before been produced, much less studied. All of these metals were observed to afford some degree of corrosion resistance to magnesium. Due to the especially promising results from nonequilibrium alloying of magnesium with yttrium and titanium, the ternary magnesium-yttrium-titanium system was investigated in depth. While all of the alloys are lustrous, surface morphology is observed under the scanning

  12. Active Nuclear Import of Membrane Proteins Revisited.

    PubMed

    Laba, Justyna K; Steen, Anton; Popken, Petra; Chernova, Alina; Poolman, Bert; Veenhoff, Liesbeth M

    2015-01-01

    It is poorly understood how membrane proteins destined for the inner nuclear membrane pass the crowded environment of the Nuclear Pore Complex (NPC). For the Saccharomyces cerevisiae proteins Src1/Heh1 and Heh2, a transport mechanism was proposed where the transmembrane domains diffuse through the membrane while the extralumenal domains encoding a nuclear localization signal (NLS) and intrinsically disordered linker (L) are accompanied by transport factors and travel through the NPC. Here, we validate the proposed mechanism and explore and discuss alternative interpretations of the data. First, to disprove an interpretation where the membrane proteins become membrane embedded only after nuclear import, we present biochemical and localization data to support that the previously used, as well as newly designed reporter proteins are membrane-embedded irrespective of the presence of the sorting signals, the specific transmembrane domain (multipass or tail anchored), independent of GET, and also under conditions that the proteins are trapped in the NPC. Second, using the recently established size limit for passive diffusion of membrane proteins in yeast, and using an improved assay, we confirm active import of polytopic membrane protein with extralumenal soluble domains larger than those that can pass by diffusion on similar timescales. This reinforces that NLS-L dependent active transport is distinct from passive diffusion. Thirdly, we revisit the proposed route through the center of the NPC and conclude that the previously used trapping assay is, unfortunately, poorly suited to address the route through the NPC, and the route thus remains unresolved. Apart from the uncertainty about the route through the NPC, the data confirm active, transport factor dependent, nuclear transport of membrane-embedded mono- and polytopic membrane proteins in baker's yeast. PMID:26473931

  13. Active Nuclear Import of Membrane Proteins Revisited

    PubMed Central

    Laba, Justyna K.; Steen, Anton; Popken, Petra; Chernova, Alina; Poolman, Bert; Veenhoff, Liesbeth M.

    2015-01-01

    It is poorly understood how membrane proteins destined for the inner nuclear membrane pass the crowded environment of the Nuclear Pore Complex (NPC). For the Saccharomyces cerevisiae proteins Src1/Heh1 and Heh2, a transport mechanism was proposed where the transmembrane domains diffuse through the membrane while the extralumenal domains encoding a nuclear localization signal (NLS) and intrinsically disordered linker (L) are accompanied by transport factors and travel through the NPC. Here, we validate the proposed mechanism and explore and discuss alternative interpretations of the data. First, to disprove an interpretation where the membrane proteins become membrane embedded only after nuclear import, we present biochemical and localization data to support that the previously used, as well as newly designed reporter proteins are membrane-embedded irrespective of the presence of the sorting signals, the specific transmembrane domain (multipass or tail anchored), independent of GET, and also under conditions that the proteins are trapped in the NPC. Second, using the recently established size limit for passive diffusion of membrane proteins in yeast, and using an improved assay, we confirm active import of polytopic membrane protein with extralumenal soluble domains larger than those that can pass by diffusion on similar timescales. This reinforces that NLS-L dependent active transport is distinct from passive diffusion. Thirdly, we revisit the proposed route through the center of the NPC and conclude that the previously used trapping assay is, unfortunately, poorly suited to address the route through the NPC, and the route thus remains unresolved. Apart from the uncertainty about the route through the NPC, the data confirm active, transport factor dependent, nuclear transport of membrane-embedded mono- and polytopic membrane proteins in baker’s yeast. PMID:26473931

  14. Polyelectrolyte multilayers impart healability to highly electrically conductive films.

    PubMed

    Li, Yang; Chen, Shanshan; Wu, Mengchun; Sun, Junqi

    2012-08-28

    Healable, electrically conductive films are fabricated by depositing Ag nanowires on water-enabled healable polyelectrolyte multilayers. The easily achieved healability of the polyelectrolyte multilayers is successfully imparted to the Ag nanowire layer. These films conveniently restore electrical conductivity lost as a result of damage by cuts several tens of micrometers wide when water is dropped on the cuts. PMID:22807199

  15. Paleoseismograms: Can Turbidite Deposits Record Flow Unsteadiness Imparted by Earthquakes?

    NASA Astrophysics Data System (ADS)

    Goldfinger, C.; Garrett, A.; Patton, J. R.; Morey, A. E.

    2012-12-01

    In paleoseismic investigations in Cascadia, Sumatra, Chile, and the Northern San Andreas Fault, we have found that it is possible to correlate individual turbidites between isolated sites using high resolution physical property data such as CT and Gamma density, P-wave velocity and magnetic susceptibility. The magnetic and density "fingerprints" of each turbidite are a reflection of grain size distributions in many lithologies. In our studies, we commonly observe "stacked" repetitions of coarse Bouma lower divisions, followed by the fining upward tail. However the long distance correlations between isolated sites and the level of detail possible are not well explained. It is difficult to explain the observation that individual events can be correlated in detail between separate channels, slope basins and onshore lakes that are not connected. To investigate this phenomenon, we have investigated several historic earthquake deposits, and have also generated turbidity currents in flume tanks to test the dynamics of transport and deposition of unsteady turbidity currents. Flume experiments commonly attempt to eliminate source heterogeneity, and assume that hydrodynamics, sediment character, and topography are the primary controls on deposition downstream. Longitudinal flow variability is rarely considered, but is predicted to have an important role in deposition (Kneller and McCaffrey, 2003), and is sometimes invoked for waxing and waning flood deposits. We question the starting assumption of no input variability, and input variable longitudinal flow regimes to simulate several common input scenarios including storm (hyperpycnal), dam breach, and long earthquake inputs. We use calibrated sediment, color coded by grain size and used photographic techniques to extract detailed grain size plots of each turbidite deposition compare to the input flow hydrograph. When compared to the input sources of the turbidity currents, we found that there is an excellent correlation

  16. Zonal drifts of irregularities imparted by meridional winds.

    NASA Technical Reports Server (NTRS)

    Waldman, H.; Da Rosa, A. V.

    1973-01-01

    In a uniform ionosphere, meridional winds cause only meridional motions of irregularities. It is shown, however, that, if F-region irregularities are considered in a real ionosphere in which there is a highly conductive E-layer, zonal motions occur. During the day a substantial westward drift takes place, while at night the drift is eastward but smaller, owing to the much smaller E-layer conductivity. Thus, the effect of meridional winds is to impart a net westward drift to small irregularities in the ionization, provided such irregularities persist long enough.

  17. IL-7 signaling imparts polyfunctionality and stemness potential to CD4(+) T cells.

    PubMed

    Ding, Zhi-Chun; Liu, Chufeng; Cao, Yang; Habtetsion, Tsadik; Kuczma, Michal; Pi, Wenhu; Kong, Heng; Cacan, Ercan; Greer, Susanna F; Cui, Yan; Blazar, Bruce R; Munn, David H; Zhou, Gang

    2016-06-01

    The functional status of CD4(+) T cells is a critical determinant of antitumor immunity. Polyfunctional CD4(+) T cells possess the ability to concomitantly produce multiple Th1-type cytokines, exhibiting a functional attribute desirable for cancer immunotherapy. However, the mechanisms by which these cells are induced are neither defined nor it is clear if these cells can be used therapeutically to treat cancer. Here, we report that CD4(+) T cells exposed to exogenous IL-7 during antigenic stimulation can acquire a polyfunctional phenotype, characterized by their ability to simultaneously express IFNγ, IL-2, TNFα and granzyme B. This IL-7-driven polyfunctional phenotype was associated with increased histone acetylation in the promoters of the effector genes, indicative of increased chromatin accessibility. Moreover, forced expression of a constitutively active (CA) form of STAT5 recapitulated IL-7 in inducing CD4(+) T-cell polyfunctionality. Conversely, the expression of a dominant negative (DN) form of STAT5 abolished the ability of IL-7 to induce polyfunctional CD4(+) T cells. These in-vitro-generated polyfunctional CD4(+) T cells can traffic to tumor and expand intratumorally in response to immunization. Importantly, adoptive transfer of polyfunctional CD4(+) T cells following lymphodepletive chemotherapy was able to eradicate large established tumors. This beneficial outcome was associated with the occurrence of antigen epitope spreading, activation of the endogenous CD8(+) T cells and persistence of donor CD4(+) T cells exhibiting memory stem cell attributes. These findings indicate that IL-7 signaling can impart polyfunctionality and stemness potential to CD4(+) T cells, revealing a previously unknown property of IL-7 that can be exploited in adoptive T-cell immunotherapy.

  18. INDIVIDUALISED CALCULATION OF TISSUE IMPARTED ENERGY IN BREAST TOMOSYNTHESIS.

    PubMed

    Geeraert, N; Klausz, R; Muller, S; Bloch, I; Bosmans, H

    2016-06-01

    The imparted energy to the glandular tissue in the breast (glandular imparted energy, GIE) is proposed for an improved assessment of the individual radiation-induced risk resulting from X-ray breast imaging. GIE is computed from an estimation of the quantity and localisation of glandular tissue in the breast. After a digital breast tomosynthesis (DBT) acquisition, the volumetric glandular content (volumetric breast density, VBD) is computed from the central X-ray projection. The glandular tissue distribution is determined by labelling the DBT voxels to ensure the conservation of the VBD. Finally, the GIE is calculated by Monte Carlo computation on the resulting tissue-labelled DBT volume. For verification, the method was applied to 10 breast-shaped digital phantoms made of different glandular spheres in an adipose background, and to a digital anthropomorphic phantom. Results were compared to direct GIE computations on the phantoms considered as 'ground-truth'. The major limitations in accuracy are those of DBT, in particular the limited z-resolution. However, for most phantoms, the results can be considered as acceptable.

  19. Ideas Exchange: "How Important Is Activity in Young Children (Preschool) to a Lifetime of Physical Activity?"

    ERIC Educational Resources Information Center

    Hushman, GLenn; Morrison, Jaime; Mally, Kristi; McCall, Renee; Corso, Marjorie; Kamla, Jim; Magnotta, John; Chase, Melissa A.; Garrahy, Deborah A.; Lorenzi, David G.; Barnd, Sue

    2009-01-01

    This article presents the opinions of several professionals who were asked: "How important is activity in young children (preschool) to a lifetime of physical activity?" These professionals point out the importance of physical activity to young children.

  20. Cytomagnetometric study of interactions between microfilaments and microtubules by measuring the energy imparted to magnetic particles within the cells

    NASA Astrophysics Data System (ADS)

    Nemoto, Iku; Kawamura, Kazuhisa

    2005-05-01

    Cytomagnetometric measurements of the energy imparted to intracellular organelles were made to study the relationship between microtubules and microfilaments. Depolymerization of microtubules by colchicine resulted in an increase in the energy suggesting that microtubules in control condition suppress the activity of microfilaments.

  1. Seasonal Pacing - Match Importance Affects Activity in Professional Soccer

    PubMed Central

    Link, Daniel; de Lorenzo, Michael F.

    2016-01-01

    This research explores the influence of match importance on player activity in professional soccer. Therefore, we used an observational approach and analyzed 1,211 matches of German Bundesliga and 2nd Bundesliga. The importance measurement employed is based on post season consequences of teams involved in a match. This means, if a match result could potentially influence the final rank, and this rank would lead to different consequences for a team, such as qualification for Champions League opposed to qualification for Europe League, then this match is classified as important; otherwise not. Activity was quantified by TOTAL DISTANCE COVERED, SPRINTS, FAST RUNS, DUELS, FOULS and ATTEMPTS. Running parameters were recorded using a semi-automatic optical tracking system, while technical variables were collected by professional data loggers. Based on our importance classification, low important matches occurred at the beginning of round 29. A two-way ANOVA indicates significantly increased FAST RUNS (+4%, d = 0.3), DUELS (+16%, d = 1.0) and FOULS (+36%, d = 1.2) in important matches compared to low important ones. For FAST RUNS and FOULS, this effect only exists in Bundesliga. A comparison of the two leagues show that TOTAL DISTANCE COVERED (+3%, d = 0.9), SPRINTS (+25%, d = 1.4) and FAST RUNS (+15%, d = 1.4) are higher compared to 2nd Bundesliga, whilst FOULS is less in Bundesliga (-7%, d = 0.3). No difference in player activity was found between matches at the beginning of a season (round 1–6) and at the end of a season (round 29–34). We conclude that match importance influences player activity in German professional soccer. The most reasonable explanation is a conscious or unconscious pacing strategy, motivated by preserving abilities or preventing injury. Since this tendency mainly exists in Bundesliga, this may suggest that more skilled players show a higher awareness for the need of pacing. PMID:27281051

  2. Seasonal Pacing - Match Importance Affects Activity in Professional Soccer.

    PubMed

    Link, Daniel; de Lorenzo, Michael F

    2016-01-01

    This research explores the influence of match importance on player activity in professional soccer. Therefore, we used an observational approach and analyzed 1,211 matches of German Bundesliga and 2nd Bundesliga. The importance measurement employed is based on post season consequences of teams involved in a match. This means, if a match result could potentially influence the final rank, and this rank would lead to different consequences for a team, such as qualification for Champions League opposed to qualification for Europe League, then this match is classified as important; otherwise not. Activity was quantified by TOTAL DISTANCE COVERED, SPRINTS, FAST RUNS, DUELS, FOULS and ATTEMPTS. Running parameters were recorded using a semi-automatic optical tracking system, while technical variables were collected by professional data loggers. Based on our importance classification, low important matches occurred at the beginning of round 29. A two-way ANOVA indicates significantly increased FAST RUNS (+4%, d = 0.3), DUELS (+16%, d = 1.0) and FOULS (+36%, d = 1.2) in important matches compared to low important ones. For FAST RUNS and FOULS, this effect only exists in Bundesliga. A comparison of the two leagues show that TOTAL DISTANCE COVERED (+3%, d = 0.9), SPRINTS (+25%, d = 1.4) and FAST RUNS (+15%, d = 1.4) are higher compared to 2nd Bundesliga, whilst FOULS is less in Bundesliga (-7%, d = 0.3). No difference in player activity was found between matches at the beginning of a season (round 1-6) and at the end of a season (round 29-34). We conclude that match importance influences player activity in German professional soccer. The most reasonable explanation is a conscious or unconscious pacing strategy, motivated by preserving abilities or preventing injury. Since this tendency mainly exists in Bundesliga, this may suggest that more skilled players show a higher awareness for the need of pacing.

  3. Seasonal Pacing - Match Importance Affects Activity in Professional Soccer.

    PubMed

    Link, Daniel; de Lorenzo, Michael F

    2016-01-01

    This research explores the influence of match importance on player activity in professional soccer. Therefore, we used an observational approach and analyzed 1,211 matches of German Bundesliga and 2nd Bundesliga. The importance measurement employed is based on post season consequences of teams involved in a match. This means, if a match result could potentially influence the final rank, and this rank would lead to different consequences for a team, such as qualification for Champions League opposed to qualification for Europe League, then this match is classified as important; otherwise not. Activity was quantified by TOTAL DISTANCE COVERED, SPRINTS, FAST RUNS, DUELS, FOULS and ATTEMPTS. Running parameters were recorded using a semi-automatic optical tracking system, while technical variables were collected by professional data loggers. Based on our importance classification, low important matches occurred at the beginning of round 29. A two-way ANOVA indicates significantly increased FAST RUNS (+4%, d = 0.3), DUELS (+16%, d = 1.0) and FOULS (+36%, d = 1.2) in important matches compared to low important ones. For FAST RUNS and FOULS, this effect only exists in Bundesliga. A comparison of the two leagues show that TOTAL DISTANCE COVERED (+3%, d = 0.9), SPRINTS (+25%, d = 1.4) and FAST RUNS (+15%, d = 1.4) are higher compared to 2nd Bundesliga, whilst FOULS is less in Bundesliga (-7%, d = 0.3). No difference in player activity was found between matches at the beginning of a season (round 1-6) and at the end of a season (round 29-34). We conclude that match importance influences player activity in German professional soccer. The most reasonable explanation is a conscious or unconscious pacing strategy, motivated by preserving abilities or preventing injury. Since this tendency mainly exists in Bundesliga, this may suggest that more skilled players show a higher awareness for the need of pacing. PMID:27281051

  4. A facile method for imparting superoleophobicity to polymer substrates

    NASA Astrophysics Data System (ADS)

    Ren, Guina; Zhang, Zhaozhu; Zhu, Xiaotao; Ge, Bo; Wang, Kun; Xu, Xianghui; Men, Xuehu; Zhou, Xiaoyan

    2014-03-01

    A new method was presented to impart polymer substrate with superoleophobic properties. Aluminum/polymer composite was created by a hot-pressing process, and rough surface textures needed to establish superoleophobicity were created by HCl etching and boiling water treatment. After surface fluorination, the surface became super-repellent towards water and several organic liquids, such as hexadecane. The effect of geometrical structure on hydrophobicity and oleophobicity was investigated, and the result showed that the synergistic action of microterraces and nanoflakes played a key role in establishing oleophobicity. A waterfall/jet test demonstrated that the obtained surface can keep its superoleophobicity after a long time exposure to water. Moreover, the obtained surface did not lose the superoleophobicity after placing it under cold condition for 7 days.

  5. Amino acids flanking the central core of Cu,Zn superoxide dismutase are important in retaining enzyme activity after autoclaving.

    PubMed

    Kumar, Arun; Randhawa, Vinay; Acharya, Vishal; Singh, Kashmir; Kumar, Sanjay

    2016-01-01

    Enzymes are known to be denatured upon boiling, although Cu,Zn superoxide dismutase of Potentilla atrosanguinea (Pot-SOD) retains significant catalytic activity even after autoclaving (heating at 121 °C at a pressure of 1.1 kg per square cm for 20 min). The polypeptide backbone of Pot-SOD consists of 152 amino acids with a central core spanning His45 to Cys145 that is involved in coordination of Cu(2+) and Zn(2+) ions. While the central core is essential for imparting catalytic activity and structural stability to the enzyme, the role of sequences flanking the central core was not understood. Experiments with deletion mutants showed that the amino acid sequences flanking the central core were important in retaining activity of Pot-SOD after autoclaving. Molecular dynamics simulations demonstrated the unfavorable structure of mutants due to increased size of binding pocket and enhanced negative charge on the electrostatic surface, resulting in unavailability of the substrate superoxide radical ([Formula: see text]) to the catalytic pocket. Deletion caused destabilization of structural elements and reduced solvent accessibility that further produced unfavorable structural geometry of the protein. PMID:25990646

  6. Parent participation plays an important part in promoting physical activity

    PubMed Central

    Lindqvist, Anna-Karin; Kostenius, Catrine; Gard, Gunvor; Rutberg, Stina

    2015-01-01

    Although physical activity (PA) is an important and modifiable determinant of health, in Sweden only 15% of boys and 10% of girls aged 15 years old achieve the recommended levels of PA 7 days per week. Adolescents’ PA levels are associated with social influence exerted by parents, friends, and teachers. The purpose of this study was to describe parents’ experiences of being a part of their adolescents’ empowerment-inspired PA intervention. A qualitative interview study was performed at a school in the northern part of Sweden. A total of 10 parents were interviewed, and the collected data were analyzed with qualitative content analysis. Three subthemes were combined into one main theme, demonstrating that parents are one important part of a successful PA intervention. The life of an adolescent has many options and demands that make it difficult to prioritize PA. Although parents felt that they were important in supporting their adolescent, a successful PA intervention must have multiple components. Moreover, the parents noted that the intervention had a positive effect upon not only their adolescents’, but also their own PA. Interventions aimed at promoting PA among adolescents should include measures to stimulate parent participation, have an empowerment approach, and preferably be school-based. PMID:26282870

  7. Stellar Activity and CMEs: Important Factors of Planetary Evolution

    NASA Astrophysics Data System (ADS)

    Khodachenko, Maxim L.

    CME activity of the Sun is known to be an important impacting factor for the magnetospheres, atmospheres, and surfaces of solar system planets. Following an idea of a solar-stellar analogy, CME phenomena are expected on other stars as well. The main planetary impact factors of the stellar CMEs include the associated interplanetary shocks, plasma density and velocity disturbances, energetic particles accelerated in the shock regions, as well as distortions of the magnetic field direction and modulus. All these factors should be properly taken into account during the study of evolutionary processes on exoplanets and their atmospheric and plasma environments. The planetary impact of the stellar CME activity may vary depending on stellar age, stellar spectral type and the orbital distance of a planet. Because of the relatively short range of propagation of the majority of CMEs, they affect most strongly the magnetospheres and atmospheres of close-orbit ( < 0.1 AU) exoplanets. In this chapter we discuss an issue of the stellar CME activity in the context of several actual problems of modern exoplanetology, including planetary atmosphere mass loss, planet survival at close orbits, and definition of a criterion for habitability.

  8. Centriolar CPAP/SAS-4 Imparts Slow Processive Microtubule Growth.

    PubMed

    Sharma, Ashwani; Aher, Amol; Dynes, Nicola J; Frey, Daniel; Katrukha, Eugene A; Jaussi, Rolf; Grigoriev, Ilya; Croisier, Marie; Kammerer, Richard A; Akhmanova, Anna; Gönczy, Pierre; Steinmetz, Michel O

    2016-05-23

    Centrioles are fundamental and evolutionarily conserved microtubule-based organelles whose assembly is characterized by microtubule growth rates that are orders of magnitude slower than those of cytoplasmic microtubules. Several centriolar proteins can interact with tubulin or microtubules, but how they ensure the exceptionally slow growth of centriolar microtubules has remained mysterious. Here, we bring together crystallographic, biophysical, and reconstitution assays to demonstrate that the human centriolar protein CPAP (SAS-4 in worms and flies) binds and "caps" microtubule plus ends by associating with a site of β-tubulin engaged in longitudinal tubulin-tubulin interactions. Strikingly, we uncover that CPAP activity dampens microtubule growth and stabilizes microtubules by inhibiting catastrophes and promoting rescues. We further establish that the capping function of CPAP is important to limit growth of centriolar microtubules in cells. Our results suggest that CPAP acts as a molecular lid that ensures slow assembly of centriolar microtubules and, thereby, contributes to organelle length control.

  9. LOV-based optogenetic devices: light-driven modules to impart photoregulated control of cellular signaling

    PubMed Central

    Pudasaini, Ashutosh; El-Arab, Kaley K.; Zoltowski, Brian D.

    2015-01-01

    The Light-Oxygen-Voltage domain family of proteins is widespread in biology where they impart sensory responses to signal transduction domains. The small, light responsive LOV modules offer a novel platform for the construction of optogenetic tools. Currently, the design and implementation of these devices is partially hindered by a lack of understanding of how light drives allosteric changes in protein conformation to activate diverse signal transduction domains. Further, divergent photocycle properties amongst LOV family members complicate construction of highly sensitive devices with fast on/off kinetics. In the present review we discuss the history of LOV domain research with primary emphasis on tuning LOV domain chemistry and signal transduction to allow for improved optogenetic tools. PMID:25988185

  10. Important biological activities induced by Thalassophryne maculosa fish venom.

    PubMed

    Sosa-Rosales, Josefina Ines; Piran-Soares, Ana Amélia; Farsky, Sandra H P; Takehara, Harumi Ando; Lima, Carla; Lopes-Ferreira, Mônica

    2005-02-01

    The accidents caused by Thalassophryne maculosa fish venoms are frequent and represent a public health problem in some regions of Venezuela. Most accidents occur in the fishing communities and tourists. The clinical picture is characterized by severe pain, dizziness, fever, edema, and necrosis. Due to the lack of efficient therapy it may take weeks, or even months for complete recovery of the victims. The investigations presented here were undertaken to assess the eletrophoretical profile and principal biological properties of the T. maculosa venom. Venom obtained from fresh captured specimens of this fish was tested in vitro or in animal models for a better characterization of its toxic activities. In contrast to other fish venoms, T. maculosa venom showed relative low LD50. The injection of venom in the footpad of mice reproduced a local inflammatory lesion similar to that described in humans. Significant increase of the nociceptive and edematogenic responses was observed followed within 48 h by necrosis. Pronounced alterations on microvascular hemodynamics were visualized after venom application. These alterations were represented by fibrin depots and thrombus formation followed by complete venular stasis and transient arteriolar contraction. T. maculosa venom is devoid of phospholipase A2 activity, but the venom showed proteolytic and myotoxic activities. SDS-Page analysis of the crude venom showed important bands: one band located above 97 M(w), one band between 68 and 97 M(w), one major band between 29 and 43 M(w) and the last one located below 18.4 M(w) Then, the results presented here support that T. maculosa venom present a mixture of bioactive toxins involved in a local inflammatory lesion. PMID:15626364

  11. Dike propagation in active volcanoes: importance, evidence, models and perspectives

    NASA Astrophysics Data System (ADS)

    Acocella, V.

    2011-12-01

    Most eruptions are fed by dikes; therefore, better knowledge of dike propagation is crucial to improve our understanding of how magma is transferred and extruded at volcanoes. Dike pattern data from a few tens of active volcanic edifices show how dike propagation in a volcano is not a random process; rather, it depends from the following factors (listed in order of importance): the presence of relief, the shape of the edifice, the proximity to the surface, and regional tectonic control. Relief enhances the development of radial dikes, which may also cluster following volcano elongation or regional patterns. Dikes approaching the surface of volcanic edifices, regardless of their initial orientation, reorient to become radial (parallel to the maximum gravitational stress); in presence of scarps, dikes reorient subparallel to the scarp (perpendicular to the minimum gravitational stress). These relationships have been also observed or inferred during eruptions at Etna, Stromboli, Vesuvio (Italy), Erta Ale (Afar) and Faial (Azores). While numerical modelling of dike propagation remains challenging, analogue models of dike emplacement have been performed over a few decades, also supporting part of the above-described evidence. Analogue models have been mostly conducted injecting air or water within gelatine and, recently, injecting vegetable oil within sand. More sophisticated analogue modelling is foreseen for the future, using a more appropriate scaling, a larger sensitivity and providing a more quantitative approach in capturing relationships. More in general, future research on dikes should be devoted towards identifying dike propagation paths, dike arrest mechanisms, and likely locations of vent formation at specific volcanoes, to better aid hazards assessment.

  12. Active Learning: The Importance of Developing a Comprehensive Measure

    ERIC Educational Resources Information Center

    Carr, Rodney; Palmer, Stuart; Hagel, Pauline

    2015-01-01

    This article reports on an investigation into the validity of a widely used scale for measuring the extent to which higher education students employ active learning strategies. The scale is the active learning scale in the Australasian Survey of Student Engagement. This scale is based on the Active and Collaborative Learning scale of the National…

  13. 75 FR 50772 - Agency Information Collection Activities: Importation Bond Structure

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-17

    ... Structure AGENCY: U.S. Customs and Border Protection, Department of Homeland Security. ] ACTION: 60-Day... concerning the: Importation Bond Structure. This request for comment is being made pursuant to the Paperwork...: Title: Importation Bond Structure. OMB Number: 1651-0050. Form Numbers: 301 and 5297. Abstract:...

  14. 78 FR 75576 - Agency Information Collection Activities: Importation Bond Structure

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-12-12

    ... Structure AGENCY: U.S. Customs and Border Protection (CBP), Department of Homeland Security. ACTION: 60-day... Importation Bond Structure. This request for comment is being made pursuant to the Paperwork Reduction Act of... Structure. OMB Number: 1651-0050. Form Number: CBP Forms 301 and 5297. Abstract: Bonds are used to...

  15. 75 FR 68809 - Agency Information Collection Activities: Importation Bond Structure

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-11-09

    ... information collection was previously published in the Federal Register (75 FR 50772) on August 17, 2010... Structure AGENCY: U.S. Customs and Border Protection, Department of Homeland Security. ACTION: 30-day notice... review and approval in accordance with the Paperwork Reduction Act: Importation Bond Structure. This is...

  16. How Important Is the Phosphatase Activity of Sensor Kinases?

    PubMed Central

    Kenney, Linda J.

    2010-01-01

    In two-component signaling systems, phosphorylated response regulators (RRs) are often dephosphorylated by their partner kinases in order to control the in vivo concentration of phospho-RR (RR~P). This activity is easily demonstrated in vitro, but these experiments have typically used very high concentrations of the histidine kinase (HK) compared to the RR~P. Many two-component systems exhibit exquisite control over the ratio of HK to RR in vivo. The question thus arises as to whether the phosphatase activity of HKs is significant in vivo. This topic will be explored in the present review. PMID:20223700

  17. The Importance of Engaging Pupils Actively in Demonstrations

    ERIC Educational Resources Information Center

    Suomela, Liisa; Juuti, Kalle; Ahtee, Maija

    2013-01-01

    Demonstrating is a traditional method in teaching science that can raise interest and encourage pupils to think about a topic. While demonstrating, the teacher can focus the pupils' attention on the relevant facts and introduce scientific principles and concepts. Through discussion and actively making observations and inferences, rather than…

  18. Compounds active against cell walls of medically important fungi.

    PubMed Central

    Hector, R F

    1993-01-01

    A number of substances that directly or indirectly affect the cell walls of fungi have been identified. Those that actively interfere with the synthesis or degradation of polysaccharide components share the property of being produced by soil microbes as secondary metabolites. Compounds specifically interfering with chitin or beta-glucan synthesis have proven effective in studies of preclinical models of mycoses, though they appear to have a restricted spectrum of coverage. Semisynthetic derivatives of some of the natural products have offered improvements in activity, toxicology, or pharmacokinetic behavior. Compounds which act on the cell wall indirectly or by a secondary mechanism of action, such as the azoles, act against diverse fungi but are usually fungistatic in nature. Overall, these compounds are attractive candidates for further development. PMID:8457977

  19. [Current views on the importance of physical activity].

    PubMed

    Berg, A; Deibert, P; Berg, A; König, D; Dickhuth, H H

    2004-07-01

    To improve the individual health profile and to solve the growing overweight problem, a long-term change in the lifestyle to one which includes an energetically balanced combination of diet and activity is essential. Physical activity and the muscles involved are the primary means by which body composition and energy turnover are regulated. A state of imbalance has decisive consequences on the development of atherogenic and inflammatory risk factors. Additionally, the aging process is significantly influenced by the long-term retention or loss of muscle mass. The Deutsche Gesellschaft für Sportmedizin und Prävention (German Society for Sports Medicine and Prevention, a registered association) offers within the concept of a therapeutic lifestyle change, an educational program for overweight adults (M.O.B.I.L.I.S.). PMID:15526658

  20. Importance of active case detection in a malaria elimination programme

    PubMed Central

    2014-01-01

    Background With the aim of eliminating malaria from Sri Lanka by 2014, the Anti-Malaria Campaign of Sri Lanka (AMC) sought the support of Tropical and Environmental Disease and Health Associates Private Limited (TEDHA), a private sector organization. In 2009, TEDHA was assigned 43 government hospitals in the district of Mannar in the Northern Province and in districts of Trincomalee, Batticaloa and Ampara in the Eastern Province to carry out malaria surveillance to complement the surveillance activities of the AMC. Passive case detection (PCD), activated passive case detection (APCD) and active case detection (ACD) for malaria have been routinely carried out in Sri Lanka. Methods The active case detection programme of TEDHA involves screening of populations irrespective of the presence of fever or any other signs or symptoms of malaria to detect infections and residual parasite carriers. ACD is done by TEDHA in a) high risk populations through mobile malaria clinics including armed forces personnel and b) pregnant females who visit antenatal clinics for asymptomatic malaria infections during the first trimester of pregnancy. Populations are selected in consultation with the Regional Malaria Officer of the AMC thus avoiding any overlap with the population screened by the government. Results TEDHA screened 387,309 individuals in the four districts for malaria by ACD including high risk groups and pregnant women between January 2010 and December 2012. During this period seven individuals were diagnosed with Plasmodium vivax infections and one individual was detected with a mixed infection of P. vivax and Plasmodium falciparum. All eight cases were detected by ACD carried out by mobile malaria clinics among high risk groups in the Mannar district. Conclusion The progress made by Sri Lanka in the malaria elimination drive is largely due to increased surveillance and judicious use of control methods which has resulted in zero indigenous malaria cases being reported since

  1. Imparting regenerative capacity to limbs by progenitor cell transplantation

    PubMed Central

    Lin, Gufa; Chen, Ying; Slack, Jonathan M.W.

    2012-01-01

    Summary The frog Xenopus can normally regenerate its limbs at early developmental stages but loses the ability during metamorphosis. This behavior provides a potential gain-of-function model for measures that can enhance limb regeneration. Here we show that frog limbs can be caused to form multidigit regenerates after receiving transplants of larval limb progenitor cells. It is necessary to activate Wnt/β -catenin signaling in the cells, and to add Sonic hedgehog, FGF10 and thymosin β4. These factors promote survival and growth of the grafted cells and also provide pattern information. The eventual regenerates are not composed solely of donor tissue; the host cells also make a substantial contribution despite their lack of regeneration-competence. Cells from adult frog legs or from regenerating tadpole tails do not promote limb regeneration, demonstrating the necessity for limb progenitor cells. These findings have obvious implications for the development of a technology to promote limb regeneration in mammals. PMID:23273877

  2. Methods of imparting information to patients in dental practice.

    PubMed

    Hein, W

    1984-03-01

    Changes are taking place in dentistry now that we know much more about the causes of caries, periodontal disease and most jaw malformations and that, through education and motivation, our patients can substantially protect themselves from these disorders. In our specialty in particular, it is true that 'prevention is better than cure'! This approach will be successful only if our patients are monitored on a regular recall basis. Attention is drawn to the existence of specific target groups. Successful results can only be achieved if individual efforts are backed by group and population prophylactic measures on the part of national or local authorities using the expertise of the dental profession. A preventively orientated practice must offer much more patient education than one which concentrates on the provision of curative services. Details of the approach to be followed should be thoroughly planned and the members of the dental team to be responsible for the tasks concerned should be identified. Appropriate equipment and working facilities are essential for the effective conduct of preventive measures. It must be emphasized that these measures in dental practice involve high costs. In attending to our patients, we should not forget that not only practical intervention but also the provision of advice is a vitally important health service.

  3. Imparting Desired Attributes by Optimization in Structural Design

    NASA Technical Reports Server (NTRS)

    Sobieszczanski-Sobieski, Jaroslaw; Venter, Gerhard

    2003-01-01

    Commonly available optimization methods typically produce a single optimal design as a Constrained minimum of a particular objective function. However, in engineering design practice it is quite often important to explore as much of the design space as possible with respect to many attributes to find out what behaviors are possible and not possible within the initially adopted design concept. The paper shows that the very simple method of the sum of objectives is useful for such exploration. By geometrical argument it is demonstrated that if every weighting coefficient is allowed to change its magnitude and its sign then the method returns a set of designs that are all feasible, diverse in their attributes, and include the Pareto and non-Pareto solutions, at least for convex cases. Numerical examples in the paper include a case of an aircraft wing structural box with thousands of degrees of freedom and constraints, and over 100 design variables, whose attributes are structural mass, volume, displacement, and frequency. The method is inherently suitable for parallel, coarse-grained implementation that enables exploration of the design space in the elapsed time of a single structural optimization.

  4. The active liquid Earth - importance of temporal and spatial variability

    NASA Astrophysics Data System (ADS)

    Arheimer, Berit

    2016-04-01

    The Planet Earth is indeed liquid and active - 71 percent of its surface is water-covered and this water never rests. Thanks to the water cycle, our planet's water supply is constantly moving from one place to another and from one form to another. Only 2.5% of the water is freshwater and it exists in the air as water vapor; it hits the ground as rain and snow; it flows on the surface from higher to lower altitudes in rivers, lakes, and glaciers; and it flows in the ground in soil, aquifers, and in all living organisms until it reaches the sea. On its way over the Earth's crust, some returns quickly to vapor again, while some is trapped and exposed to many "fill and spill" situations for a long journey. The variability in the water balance is crucial for hydrological understanding and modelling. The water cycle may appear simple, but magnitudes and rates in fluxes are very different from one place to another, resulting from variable drivers such as solar energy, precipitation and gravity in co-evolution with geology, soil, vegetation and fauna. The historical evolution, the temporal fluxes and diversity in space continue to fascinate hydrological scientists. Specific physical processes may be well known, but their boundary conditions, interactions and rate often remain unknown at a specific site and are difficult to monitor in nature. This results in mysterious features where trends in drivers do not match runoff, like the Sahelian Paradox or discharge to the Arctic Ocean. Humans have always interfered with the water cycle and engineering is fundamental for water regulation and re-allocation. Some 80% of the river flow from the northern part of the Earth is affected by fragmentation of the river channels by dams. In water management, there is always a tradeoff between upstream and downstream activities, not only regarding total water quantities but also for temporal patterns and water quality aspects. Sharing a water resource can generate conflicts but geopolitical

  5. Machine imparting complex rotary motion for lapping a spherical inner diameter

    DOEpatents

    Carroll, T.A.; Yetter, H.H.

    1985-01-30

    An apparatus for imparting complex rotary motion is used to lap an inner spherical diameter surface of a workpiece. A lapping tool consists of a dome and rod mounted along the dome's vertical axis. The workpiece containing the lapping tool is held in a gimbal which uses power derived from a secondary takeoff means to impart rotary motion about a horizontal axis. The gimbal is rotated about a vertical axis by a take means while mounted at a radially outward position on a rotating arm.

  6. Machine imparting complex rotary motion for lapping a spherical inner diameter

    DOEpatents

    Carroll, Thomas A.; Yetter, Harold H.

    1986-01-01

    An apparatus for imparting complex rotary motion is used to lap an inner spherical diameter surface of a workpiece. A lapping tool consists of a dome and rod mounted along the dome's vertical axis. The workpiece containing the lapping tool is held in a gimbal which uses power derived from a secondary takeoff means to impart rotary motion about a horizontal axis. The gimbal is rotated about a vertical axis by a take means while mounted at a radially outward position on a rotating arm.

  7. Imparting Barely Visible Impact Damage to a Stitched Composite Large-Scale Pressure Box

    NASA Technical Reports Server (NTRS)

    Lovejoy, Andrew E.; Przekop, Adam

    2016-01-01

    The Pultruded Rod Stitched Efficient Unitized Structure (PRSEUS) is a concept that was developed by The Boeing Company to address the complex structural design aspects associated with a pressurized hybrid wing body (HWB) aircraft configuration, which has been a focus of the NASA Environmentally Responsible Aviation Project. The NASA-Boeing structural development for the HWB aircraft culminated in testing of the multi-bay box, which is an 80%-scale representation of the pressurized center-body section. This structure was tested in the NASA Langley Research Center Combined Loads Test System facility. As part of this testing, barely visible impact damage was imparted to the interior and exterior of the test article to demonstrate compliance with a condition representative of the requirements for Category 1 damaged composite structure as defined by the Federal Aviation Regulations. Interior impacts were imparted using an existing spring-loaded impactor, while the exterior impacts were imparted using a newly designed, gravity-driven impactor. This paper describes the impacts to the test article, and the design of the gravitydriven guided-weight impactor. The guided-weight impactor proved to be a very reliable method to impart barely visible impact damage in locations which are not easily accessible for a traditional drop-weight impactor, while at the same time having the capability to be highly configurable for use on other aircraft structures.

  8. Imparting small vorticity to a Bianchi type-VIh empty spacetime

    NASA Astrophysics Data System (ADS)

    Batakis, Nikos A.

    1981-04-01

    We present and briefly discuss a Bianchi type-VIh empty spacetime. The field equations have been solved after being linearized with respect to a parameter which imparts vorticity to the model. The limit of zero vorticity is an already known solution.

  9. Imparting Social Capital to Educationally Disadvantaged Students: A Study of the Early Academic Outreach Program

    ERIC Educational Resources Information Center

    Jackson, Nicole Korgie

    2010-01-01

    The purpose of this study is to evaluate the process of an academic preparation program, also known as an outreach program, imparting social capital to educationally disadvantaged students. The particular form of social capital to be examined will be the college knowledge needed to successfully prepare for college admission. The theoretical…

  10. Effectiveness of Different Medium of Education to Imparting Knowledge at Bangladesh Open University

    ERIC Educational Resources Information Center

    Islam, Anwarul; Islam, Nasirul

    2008-01-01

    Open and distance learning system meanly based on different types of media to impart education to the learners. Bangladesh Open University (BOU) offered education through open and distance learning system. There are two largest programs one is Secondary School Certificate (SSC) and another one is Higher Secondary Certificate (HSC) offered by BOU…

  11. Imparting Catalyst-Control upon Classical Palladium-Catalyzed Alkenyl C–H Bond Functionalization Reactions

    PubMed Central

    Sigman, Matthew S.; Werner, Erik W.

    2011-01-01

    Conspectus The functional group transformations carried out by the palladium-catalyzed Wacker and Heck reactions are radically different, but they are both alkenyl C-H bond functionalization reactions that have found extensive use in organic synthesis. The synthetic community depends heavily on these important reactions, but selectivity issues arising from control by the substrate, rather than control by the catalyst, have prevented the realization of their full potential. Because of important similarities in the respective selectivity-determining nucleopalladation and β-hydride elimination steps of these processes, we posit that the mechanistic insight garnered through the development of one of these catalytic reactions may be applied to the other. In this Account, we detail our efforts to develop catalyst-controlled variants of both the Wacker oxidation and the Heck reaction to address synthetic limitations and provide mechanistic insight into the underlying organometallic processes of these reactions. In contrast to previous reports, we discovered that electrophilic palladium catalysts with non-coordinating counterions allowed for the use of a Lewis basic ligand to efficiently promote TBHP-mediated Wacker oxidation reactions of styrenes. This discovery led to the mechanistically guided development of a Wacker reaction catalyzed by a palladium complex with a bidentate ligand. This ligation may prohibit coordination of allylic heteroatoms, thereby allowing for the application of the Wacker oxidation to substrates that were poorly behaved under classical conditions. Likewise, we unexpectedly discovered that electrophilic Pd-σ-alkyl intermediates are capable of distinguishing between electronically inequivalent C–H bonds during β-hydride elimination. As a result, we have developed E-styrenyl selective oxidative Heck reactions of previously unsuccessful electronically non-biased alkene substrates using arylboronic acid derivatives. The mechanistic insight gained

  12. Imparting Motion to a Test Object Such as a Motor Vehicle in a Controlled Fashion

    NASA Technical Reports Server (NTRS)

    Southward, Stephen C. (Inventor); Reubush, Chandler (Inventor); Pittman, Bryan (Inventor); Roehrig, Kurt (Inventor); Gerard, Doug (Inventor)

    2014-01-01

    An apparatus imparts motion to a test object such as a motor vehicle in a controlled fashion. A base has mounted on it a linear electromagnetic motor having a first end and a second end, the first end being connected to the base. A pneumatic cylinder and piston combination have a first end and a second end, the first end connected to the base so that the pneumatic cylinder and piston combination is generally parallel with the linear electromagnetic motor. The second ends of the linear electromagnetic motor and pneumatic cylinder and piston combination being commonly linked to a mount for the test object. A control system for the linear electromagnetic motor and pneumatic cylinder and piston combination drives the pneumatic cylinder and piston combination to support a substantial static load of the test object and the linear electromagnetic motor to impart controlled motion to the test object.

  13. Design and Use of a Guided Weight Impactor to Impart Barely Visible Impact Damage

    NASA Technical Reports Server (NTRS)

    Lovejoy, Andrew E.; Przekop, Adam

    2016-01-01

    Aircraft structure is required to demonstrate satisfaction of the FAR requirements for Category 1, such as barely visible impact damage (BVID). Typical aircraft structure is impacted using a dropped weight impactor, which can impart BVID to the top surface of the structure. A recent test of a multi-bay box (MBB) composite test article, that represents an 80% scale center section of a hybrid wing body aircraft, required impact to be in a direction other than vertical from above, but still in an direction that is normal to the surface. This requirement eliminated the use of the conventional dropped weight impactor. Therefore, a design study was undertaken to determine the most effective way to efficiently and reliably impact the MBB. The chosen design was a guided weight impactor that is gravity driven. This paper describes the design of the guided weight impactor, and presents the results of its use for imparting BVID to the MBB. The guided weight impactor was seen to be a very reliable method to impart BVID, while at the same time having the capability to be highly configurable for use on other aircraft structure that is impacted at a variety of impact energies and from a variety of directions.

  14. Model for radial dependence of frequency distributions for energy imparted in nanometer volumes from HZE particles

    NASA Technical Reports Server (NTRS)

    Cucinotta, F. A.; Nikjoo, H.; Goodhead, D. T.; Decillo, J. F. (Principal Investigator)

    2000-01-01

    This paper develops a deterministic model of frequency distributions for energy imparted (total energy deposition) in small volumes similar to DNA molecules from high-energy ions of interest for space radiation protection and cancer therapy. Frequency distributions for energy imparted are useful for considering radiation quality and for modeling biological damage produced by ionizing radiation. For high-energy ions, secondary electron (delta-ray) tracks originating from a primary ion track make dominant contributions to energy deposition events in small volumes. Our method uses the distribution of electrons produced about an ion's path and incorporates results from Monte Carlo simulation of electron tracks to predict frequency distributions for ions, including their dependence on radial distance. The contribution from primary ion events is treated using an impact parameter formalism of spatially restricted linear energy transfer (LET) and energy-transfer straggling. We validate our model by comparing it directly to results from Monte Carlo simulations for proton and alpha-particle tracks. We show for the first time frequency distributions of energy imparted in DNA structures by several high-energy ions such as cosmic-ray iron ions. Our comparison with results from Monte Carlo simulations at low energies indicates the accuracy of the method.

  15. Model for radial dependence of frequency distributions for energy imparted in nanometer volumes from HZE particles.

    PubMed

    Cucinotta, F A; Nikjoo, H; Goodhead, D T

    2000-04-01

    This paper develops a deterministic model of frequency distributions for energy imparted (total energy deposition) in small volumes similar to DNA molecules from high-energy ions of interest for space radiation protection and cancer therapy. Frequency distributions for energy imparted are useful for considering radiation quality and for modeling biological damage produced by ionizing radiation. For high-energy ions, secondary electron (delta-ray) tracks originating from a primary ion track make dominant contributions to energy deposition events in small volumes. Our method uses the distribution of electrons produced about an ion's path and incorporates results from Monte Carlo simulation of electron tracks to predict frequency distributions for ions, including their dependence on radial distance. The contribution from primary ion events is treated using an impact parameter formalism of spatially restricted linear energy transfer (LET) and energy-transfer straggling. We validate our model by comparing it directly to results from Monte Carlo simulations for proton and alpha-particle tracks. We show for the first time frequency distributions of energy imparted in DNA structures by several high-energy ions such as cosmic-ray iron ions. Our comparison with results from Monte Carlo simulations at low energies indicates the accuracy of the method. PMID:10761008

  16. Physical activities and their importance to the health of people with severe mental illness in Sweden.

    PubMed

    Erdner, Anette; Magnusson, Annabella

    2012-10-01

    It is well known that people with severe mental illness often suffer from constant fatigue, insomnia, and somatic complaints that are too often overlooked. In addition, these persons die earlier in life than others in the population. The purpose of this study was to investigate patients' descriptions of activities and the importance of these activities for their health. Eight persons living in their own home were interviewed about both their views about exercise and their exercising activities. Two themes emerged: Getting Control over One's Life and The Need for Contact with Family & Friends. All of the informants were aware of the importance of physical activity to feel good. The informants described three different forms of activities: daily activities in the home, activities in a rehabilitation centre, and various forms of jogging. These different forms of activity were important to the informants since they reduced their anxiety and stress. PMID:23017044

  17. 77 FR 37696 - Agency Information Collection Activities: Importer ID Input Record

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-06-22

    ... SECURITY U.S. Customs and Border Protection Agency Information Collection Activities: Importer ID Input... other Federal agencies to comment on an information collection requirement concerning the Importer ID... concerning the following information collection: Title: Importer ID Input Record. OMB Number: 1651-0064....

  18. 78 FR 67383 - Agency Information Collection Activities; Importer ID Input Record

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-11-12

    ... SECURITY U.S. Customs and Border Protection Agency Information Collection Activities; Importer ID Input... other Federal agencies to comment on an information collection requirement concerning the Importer ID... concerning the following information collection: Title: Importer ID Input Record. OMB Number: 1651-0064....

  19. Mapping neural circuits with activity-dependent nuclear import of a transcription factor.

    PubMed

    Masuyama, Kaoru; Zhang, Yi; Rao, Yi; Wang, Jing W

    2012-03-01

    Abstract: Nuclear factor of activated T cells (NFAT) is a calcium-responsive transcription factor. We describe here an NFAT-based neural tracing method-CaLexA (calcium-dependent nuclear import of LexA)-for labeling active neurons in behaving animals. In this system, sustained neural activity induces nuclear import of the chimeric transcription factor LexA-VP16-NFAT, which in turn drives green fluorescent protein (GFP) reporter expression only in active neurons. We tested this system in Drosophila and found that volatile sex pheromones excite specific neurons in the olfactory circuit. Furthermore, complex courtship behavior associated with multi-modal sensory inputs activated neurons in the ventral nerve cord. This method harnessing the mechanism of activity-dependent nuclear import of a transcription factor can be used to identify active neurons in specific neuronal population in behaving animals. PMID:22236090

  20. The importance of space policy teaching in communicating space activities to society [rapid communication

    NASA Astrophysics Data System (ADS)

    Reibaldi, G. G.

    2003-12-01

    The governments' priority and budgets for space activities are steadily decreasing and the importance of space activities is not any longer reaching the front pages of the newspaper, as in the 1960s. On the other hand in Europe the people, at large, have shown an important interest and support for space activities. A contribution to bridge the gap between decreasing funding and important support of citizen can come from teaching space policy in universities as well as in special workshops for government, industrial and military circles. The paper will outline a course that fulfils this goal.

  1. Imparting the unique properties of DNA into complex material architectures and functions

    PubMed Central

    Xu, Phyllis F.; Noh, Hyunwoo; Lee, Ju Hun; Domaille, Dylan W.; Nakatsuka, Matthew A.; Goodwin, Andrew P.; Cha, Jennifer N.

    2014-01-01

    While the remarkable chemical and biological properties of DNA have been known for decades, these properties have only been imparted into materials with unprecedented function much more recently. The inimitable ability of DNA to form programmable, complex assemblies through stable, specific, and reversible molecular recognition has allowed the creation of new materials through DNA’s ability to control a material’s architecture and properties. In this review we discuss recent progress in how DNA has brought unmatched function to materials, focusing specifically on new advances in delivery agents, devices, and sensors. PMID:25525408

  2. Contractile activity-induced adaptations in the mitochondrial protein import system.

    PubMed

    Takahashi, M; Chesley, A; Freyssenet, D; Hood, D A

    1998-05-01

    We previously demonstrated that subsarcolemmal (SS) and intermyofibrillar (IMF) mitochondrial subfractions import proteins at different rates. This study was undertaken to investigate 1) whether protein import is altered by chronic contractile activity, which induces mitochondrial biogenesis, and 2) whether these two subfractions adapt similarly. Using electrical stimulation (10 Hz, 3 h/day for 7 and 14 days) to induce contractile activity, we observed that malate dehydrogenase import into the matrix of the SS and IMF mitochondia isolated from stimulated muscle was significantly increased by 1.4-to 1.7-fold, although the pattern of increase differed for each subfraction. This acceleration of import may be mitochondrial compartment specific, since the import of Bcl-2 into the outer membrane was not affected. Contractile activity also modified the mitochondrial content of proteins comprising the import machinery, as evident from increases in the levels of the intramitochondrial chaperone mtHSP70 as well as the outer membrane import receptor Tom20 in SS and IMF mitochondria. Addition of cytosol isolated from stimulated or control muscles to the import reaction resulted in similar twofold increases in the ability of mitochondria to import malate dehydrogenase, despite elevations in the concentration of mitochondrial import-stimulating factor within the cytosol of chronically stimulated muscle. These results suggest that chronic contractile activity modifies the extra- and intramitochondrial environments in a fashion that favors the acceleration of precursor protein import into the matrix of the organelle. This increase in protein import is likely an important adaptation in the overall process of mitochondrial biogenesis. PMID:9612226

  3. Physical Activity in the Heat: Important Considerations to Keep Your Students Safe

    ERIC Educational Resources Information Center

    Roetert, E. Paul; Richardson, Cheryl L.; Bergeron, Michael F.

    2012-01-01

    Although July and August tend to be the warmest months of the year, the months leading up to summer as well as the months just following summer can also be quite warm or even very hot. In this article, the authors share some important information to help prepare physical educators for overseeing activities in the heat and, just as importantly, to…

  4. Alanine-Scanning Mutational Analysis of Durancin GL Reveals Residues Important for Its Antimicrobial Activity.

    PubMed

    Ju, Xingrong; Chen, Xinquan; Du, Lihui; Wu, Xueyou; Liu, Fang; Yuan, Jian

    2015-07-22

    Durancin GL is a novel class IIa bacteriocin with 43 residues produced by Enterococcus durans 41D. This bacteriocin demonstrates narrow inhibition spectrum and potent antimicrobial activity against several Listeria monocytogenes strains, including nisin-resistant L. monocytogenes NR30. A systematic alanine-scanning mutational analysis with site-directed mutagenesis was performed to analyze durancin GL residues important for antimicrobial activity and specificity. Results showed that three mutations lost their antimicrobial activity, ten mutations demonstrated a decreased effect on the activity, and seven mutations exhibited relatively high activity. With regard to inhibitory spectrum, four mutants demonstrated a narrower antimicrobial spectrum than wild-type durancin GL. Another four mutants displayed a broader target cell spectrum and increased potency relative to wild-type durancin GL. These findings broaden our understanding of durancin GL residues important for its antimicrobial activity and contribute to future rational design of variants with increased potency.

  5. Alanine-Scanning Mutational Analysis of Durancin GL Reveals Residues Important for Its Antimicrobial Activity.

    PubMed

    Ju, Xingrong; Chen, Xinquan; Du, Lihui; Wu, Xueyou; Liu, Fang; Yuan, Jian

    2015-07-22

    Durancin GL is a novel class IIa bacteriocin with 43 residues produced by Enterococcus durans 41D. This bacteriocin demonstrates narrow inhibition spectrum and potent antimicrobial activity against several Listeria monocytogenes strains, including nisin-resistant L. monocytogenes NR30. A systematic alanine-scanning mutational analysis with site-directed mutagenesis was performed to analyze durancin GL residues important for antimicrobial activity and specificity. Results showed that three mutations lost their antimicrobial activity, ten mutations demonstrated a decreased effect on the activity, and seven mutations exhibited relatively high activity. With regard to inhibitory spectrum, four mutants demonstrated a narrower antimicrobial spectrum than wild-type durancin GL. Another four mutants displayed a broader target cell spectrum and increased potency relative to wild-type durancin GL. These findings broaden our understanding of durancin GL residues important for its antimicrobial activity and contribute to future rational design of variants with increased potency. PMID:26168032

  6. Importance of Endosomal Cathelicidin Degradation To Enhance DNA-Induced Chicken Macrophage Activation.

    PubMed

    Coorens, Maarten; van Dijk, Albert; Bikker, Floris; Veldhuizen, Edwin J A; Haagsman, Henk P

    2015-10-15

    Cathelicidins are essential in the protection against invading pathogens through both their direct antimicrobial activity and their immunomodulatory functions. Although cathelicidins are known to modulate activation by several TLR ligands, little is known about their influence on DNA-induced macrophage activation. In this study, we explored the effects of cathelicidins on DNA-induced activation of chicken macrophages and elucidated the intracellular processes underlying these effects. Our results show that chicken cathelicidin (CATH)-2 strongly enhances DNA-induced activation of both chicken and mammalian macrophages because of enhanced endocytosis of DNA-CATH-2 complexes. After endocytosis, DNA is liberated from the complex because of proteolytic breakdown of CATH-2, after which TLR21 is activated. This leads to increased cytokine expression and NO production. Through the interaction with DNA, CATH-2 can play an important role in modulating the immune response at sites of infection. These observations underline the importance of cathelicidins in sensing bacterial products and regulating immune responses.

  7. An automated technique for estimating patient-specific regional imparted energy and dose in TCM CT exams

    NASA Astrophysics Data System (ADS)

    Sanders, Jeremiah W.; Tian, Xiaoyu; Segars, W. Paul; Boone, John; Samei, Ehsan

    2016-03-01

    Currently computed tomography (CT) dosimetry relies on CT dose index (CTDI) and size specific dose estimates (SSDE). Organ dose is a better metric of radiation burden. However, organ dose estimation requires precise knowledge of organ locations. Regional imparted energy and dose can also be used to quantify radiation burden. Estimating the imparted energy from CT exams is beneficial in that it does not require precise estimates of the organ size or location. This work investigated an automated technique for retrospectively estimating the imparted energy from chest and abdominopelvic tube current modulated (TCM) CT exams. Monte Carlo simulations of chest and abdominopelvic TCM CT examinations across various tube potentials and TCM strengths were performed on 58 adult computational extended cardiac-torso (XCAT) phantoms to develop relationships between scanned mass and imparted energy normalized by dose length product (DLP). An automated algorithm for calculating the scanned patient volume was further developed using an open source mesh generation toolbox. The scanned patient volume was then used to estimate the scanned mass accounting for diverse density within the scan region. The scanned mass and DLP from the exam were used to estimate the imparted energy to the patient using the knowledgebase developed from the Monte Carlo simulations. Patientspecific imparted energy estimates were made from 20 chest and 20 abdominopelvic clinical CT exams. The average imparted energy was 274 +/- 141 mJ and 681 +/- 376 mJ for the chest and abdominopelvic exams, respectively. This method can be used to estimate the regional imparted energy and/or regional dose in chest and abdominopelvic TCM CT exams across clinical operations.

  8. Heat-shock stress activates a novel nuclear import pathway mediated by Hikeshi

    PubMed Central

    Imamoto, Naoko; Kose, Shingo

    2012-01-01

    Cellular stresses significantly affect nuclear transport systems. Nuclear transport pathways mediated by importin β-family members, which are active under normal conditions, are downregulated. During thermal stress, a nuclear import pathway mediated by a novel carrier, which we named Hikeshi, becomes active. Hikeshi is not a member of the importin β family and mediates the nuclear import of Hsp70s. Unlike importin β family-mediated nuclear transport, the Hikeshi-mediated nuclear import of Hsp70s is not coupled to the GTPase cycle of the small GTPase Ran but rather is coupled with the ATPase cycle of Hsp70s. Hikeshi-mediated nuclear import is essential for the attenuation and reversal of the thermal stress response in human cells. The mechanism and functions of this newly identified nuclear import pathway will be discussed. PMID:22895094

  9. The importance of active efflux systems in the quinolone resistance of clinical isolates of Salmonella spp.

    PubMed

    Escribano, Isabel; Rodríguez, Juan Carlos; Cebrian, Laura; Royo, Gloria

    2004-11-01

    The aim of this study was to determine the importance of the active elimination of antibiotics by active efflux systems, in the decrease in fluoroquinolone sensitivity of clinical isolates of Salmonella spp. as well as the intrinsic antibiotic activity of certain active efflux system inhibitors. The effect of the active efflux system on the decrease in sensitivity to nalidixic acid, ciprofloxacin, ofloxacin and sparfloxacin was studied by investigating the variation in the in vitro activity of these compounds when assayed in association with reserpine and MC 207.110. The active efflux systems inhibited by reserpine displayed low activity in the elimination of these compounds, whereas those inhibited by MC 207.110 showed high activity in the elimination of nalidixic acid and sparfloxacin, but were less effective in the elimination of ofloxacin and ciprofloxacin. These two compounds did not exhibit intrinsic inhibitory activity against Salmonella spp. at the concentrations assayed. These mechanisms of resistance to antibiotics are complex and vary depending on the chemical composition of the antibiotics used, and perhaps the inhibitors of these systems, although they do not exhibit any intrinsic antibiotic activity, may be used as adjuvants to increase the activity of certain antibiotics. These mechanisms complement the mutations in the gyrA gene and this supports the thesis that it is necessary to lower the breakpoint established by the NCCLS for ciprofloxacin, since the strains studied have resistance mechanisms that reduce the activity of this drug and may favour the emergence of resistant mutants during treatment.

  10. Caging of plumbagin on silver nanoparticles imparts selectivity and sensitivity to plumbagin for targeted cancer cell apoptosis.

    PubMed

    Duraipandy, N; Lakra, Rachita; Kunnavakkam Vinjimur, Srivatsan; Samanta, Debasis; K, Purna Sai; Kiran, Manikantan Syamala

    2014-11-01

    Plumbagin is a nutraceutical with potent anti-cancer activity. However, the therapeutic efficacy of plumbagin is overshadowed by the lack of sensitivity and selectivity towards cancer cells. The present study evaluated the use of nano-biotechnological intervention to cage plumbagin in silver nanoparticles for selective targeting of its biological effects towards cancerous cells. Caging of plumbagin in silver nanoparticles imparted selectivity and sensitivity to plumbagin for selective killing of cancer cells by altering the redox signalling events in the cancer cells. The selectivity and sensitivity of plumbagin towards cancer cells was due to the cumulative expression of the properties of plumbagin and nanoparticles which specifically affected the differential cancer cell microenvironment by altering the pyruvate kinase activity that regulates the ROS challenge in cancerous cells. The positive surface charge of plumbagin caged silver nanoparticles (PCSN) aids in getting them targeted towards anionic cancerous cells due to the exposed terminal carboxyl group of sialic acid residues. Furthermore, we observed that the effective concentration of the drug required to induce apoptosis was brought down to 50% upon caging of plumbagin on silver nanoparticles. We observed no such effect with the individual compound alone. The results indicated that the physico-chemical and biochemical properties of plumbagin significantly changed after conjugation with nanomaterials that facilitated "adding-in" therapeutical values to plumbagin which would otherwise be overshadowed by its lack of sensitivity and selectivity towards cancer cells. PMID:25188862

  11. Imparting functionality to biocatalysts via embedding enzymes into nanoporous materials by a de novo approach: size-selective sheltering of catalase in metal-organic framework microcrystals.

    PubMed

    Shieh, Fa-Kuen; Wang, Shao-Chun; Yen, Chia-I; Wu, Chang-Cheng; Dutta, Saikat; Chou, Lien-Yang; Morabito, Joseph V; Hu, Pan; Hsu, Ming-Hua; Wu, Kevin C-W; Tsung, Chia-Kuang

    2015-04-01

    We develop a new concept to impart new functions to biocatalysts by combining enzymes and metal-organic frameworks (MOFs). The proof-of-concept design is demonstrated by embedding catalase molecules into uniformly sized ZIF-90 crystals via a de novo approach. We have carried out electron microscopy, X-ray diffraction, nitrogen sorption, electrophoresis, thermogravimetric analysis, and confocal microscopy to confirm that the ~10 nm catalase molecules are embedded in 2 μm single-crystalline ZIF-90 crystals with ~5 wt % loading. Because catalase is immobilized and sheltered by the ZIF-90 crystals, the composites show activity in hydrogen peroxide degradation even in the presence of protease proteinase K.

  12. Tasco(®), a product of Ascophyllum nodosum, imparts thermal stress tolerance in Caenorhabditis elegans.

    PubMed

    Kandasamy, Saveetha; Fan, Di; Sangha, Jatinder Singh; Khan, Wajahatullah; Evans, Franklin; Critchley, Alan T; Prithiviraj, Balakrishnan

    2011-01-01

    Tasco(®), a commercial product manufactured from the brown alga Ascophyllum nodosum, has been shown to impart thermal stress tolerance in animals. We investigated the physiological, biochemical and molecular bases of this induced thermal stress tolerance using the invertebrate animal model, Caenorhabiditis elegans. Tasco(®) water extract (TWE) at 300 μg/mL significantly enhanced thermal stress tolerance as well as extended the life span of C. elegans. The mean survival rate of the model animals under thermal stress (35 °C) treated with 300 μg/mL and 600 μg/mL TWE, respectively, was 68% and 71% higher than the control animals. However, the TWE treatments did not affect the nematode body length, fertility or the cellular localization of daf-16. On the contrary, TWE under thermal stress significantly increased the pharyngeal pumping rate in treated animals compared to the control. Treatment with TWE also showed differential protein expression profiles over control following 2D gel-electrophoresis analysis. Furthermore, TWE significantly altered the expression of at least 40 proteins under thermal stress; among these proteins 34 were up-regulated while six were down-regulated. Mass spectroscopy analysis of the proteins altered by TWE treatment revealed that these proteins were related to heat stress tolerance, energy metabolism and a muscle structure related protein. Among them heat shock proteins, superoxide dismutase, glutathione peroxidase, aldehyde dehydrogenase, saposin-like proteins 20, myosin regulatory light chain 1, cytochrome c oxidase RAS-like, GTP-binding protein RHO A, OS were significantly up-regulated, while eukaryotic translation initiation factor 5A-1 OS, 60S ribosomal protein L18 OS, peroxiredoxin protein 2 were down regulated by TWE treatment. These results were further validated by gene expression and reporter gene expression analyses. Overall results indicate that the water soluble components of Tasco(®) imparted thermal stress tolerance in

  13. Tasco®, a Product of Ascophyllum nodosum, Imparts Thermal Stress Tolerance in Caenorhabditis elegans

    PubMed Central

    Kandasamy, Saveetha; Fan, Di; Sangha, Jatinder Singh; Khan, Wajahatullah; Evans, Franklin; Critchley, Alan T.; Prithiviraj, Balakrishnan

    2011-01-01

    Tasco®, a commercial product manufactured from the brown alga Ascophyllum nodosum, has been shown to impart thermal stress tolerance in animals. We investigated the physiological, biochemical and molecular bases of this induced thermal stress tolerance using the invertebrate animal model, Caenorhabiditis elegans. Tasco® water extract (TWE) at 300 μg/mL significantly enhanced thermal stress tolerance as well as extended the life span of C. elegans. The mean survival rate of the model animals under thermal stress (35 °C) treated with 300 μg/mL and 600 μg/mL TWE, respectively, was 68% and 71% higher than the control animals. However, the TWE treatments did not affect the nematode body length, fertility or the cellular localization of daf-16. On the contrary, TWE under thermal stress significantly increased the pharyngeal pumping rate in treated animals compared to the control. Treatment with TWE also showed differential protein expression profiles over control following 2D gel-electrophoresis analysis. Furthermore, TWE significantly altered the expression of at least 40 proteins under thermal stress; among these proteins 34 were up-regulated while six were down-regulated. Mass spectroscopy analysis of the proteins altered by TWE treatment revealed that these proteins were related to heat stress tolerance, energy metabolism and a muscle structure related protein. Among them heat shock proteins, superoxide dismutase, glutathione peroxidase, aldehyde dehydrogenase, saposin-like proteins 20, myosin regulatory light chain 1, cytochrome c oxidase RAS-like, GTP-binding protein RHO A, OS were significantly up-regulated, while eukaryotic translation initiation factor 5A-1 OS, 60S ribosomal protein L18 OS, peroxiredoxin protein 2 were down regulated by TWE treatment. These results were further validated by gene expression and reporter gene expression analyses. Overall results indicate that the water soluble components of Tasco® imparted thermal stress tolerance in the C

  14. Modulation of histone deacetylase 6 (HDAC6) nuclear import and tubulin deacetylase activity through acetylation.

    PubMed

    Liu, Yuanjing; Peng, Lirong; Seto, Edward; Huang, Suming; Qiu, Yi

    2012-08-17

    The reversible acetylation of histones and non-histone proteins by histone acetyltransferases and deacetylases (HDACs) plays a critical role in many cellular processes in eukaryotic cells. HDAC6 is a unique histone deacetylase with two deacetylase domains and a C-terminal zinc finger domain. HDAC6 resides mainly in the cytoplasm and regulates many important biological processes, including cell migration and degradation of misfold proteins. HDAC6 has also been shown to localize in the nucleus to regulate transcription. However, how HDAC6 shuttles between the nucleus and cytoplasm is largely unknown. In addition, it is not clear how HDAC6 enzymatic activity is modulated. Here, we show that HDAC6 can be acetylated by p300 on five clusters of lysine residues. One cluster (site B) of acetylated lysine is in the N-terminal nuclear localization signal region. These lysine residues in site B were converted to glutamine to mimic acetylated lysines. The mutations significantly reduced HDAC6 tubulin deacetylase activity and further impaired cell motility, but had no effect on histone deacetylase activity. More interestingly, these mutations retained HDAC6 in the cytoplasm by blocking the interaction with the nuclear import protein importin-α. The retention of HDAC6 in the cytoplasm by acetylation eventually affects histone deacetylation. Thus, we conclude that acetylation is an important post-translational modification that regulates HDAC6 tubulin deacetylase activity and nuclear import.

  15. 77 FR 55486 - Agency Information Collection Activities: Importer ID Input Record

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-09-10

    ... collection techniques or the use of other forms of information technology; and (e) the annual costs burden to... SECURITY U.S. Customs and Border Protection Agency Information Collection Activities: Importer ID Input... and request for comments; Extension of an existing collection of information. SUMMARY: U.S....

  16. How My Daughter Taught Me to Teach: The Importance of Active Communication

    ERIC Educational Resources Information Center

    Hunt-Gierut, Deborah

    2011-01-01

    In this article, the author shares how her daughter, who was diagnosed with a profound hearing loss when she was a year old, taught her to teach, and demonstrates the importance of active communication. Teaching her daughter English as her second language has posed many challenges, but has also revealed successful strategies that the author has…

  17. 78 FR 44572 - Draft Guidance for Industry on Pre-Launch Activities Importation Requests; Availability

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-07-24

    ... HUMAN SERVICES Food and Drug Administration Draft Guidance for Industry on Pre-Launch Activities Importation Requests; Availability AGENCY: Food and Drug Administration, HHS. ACTION: Notice. SUMMARY: The Food and Drug Administration (FDA) is announcing the availability of a draft guidance for...

  18. The importance of domain closure for the auto-activation of ERK2

    PubMed Central

    Barr, Daniel; Oashi, Taiji; Burkhard, Kimberly; Lucius, Sarah; Samadani, Ramin; Zhang, Jun; Shapiro, Paul; MacKerell, Alexander D.; van der Vaart, Arjan

    2011-01-01

    Extracellular signal-regulated kinases-1 and 2 (ERK1/2) play a critical role in regulating cell division and have been implicated in cancer. In addition to activation by the MAPK/ERK kinases 1 and 2 (MEK1/2), certain mutants of ERK2 can be activated by auto-phosphorylation. To identify the mechanism of auto-activation, we have performed a series of molecular dynamics simulations of ERK1/2 in various stages of activation as well as the constitutively active Q103A, I84A, L73P and R65S ERK2 mutants. Our simulations indicate the importance of domain closure for auto-activation and activity regulation, with that event occurring prior to folding of the activation lip and of loop L16. Results indicate that the second phosphorylation event to T183 disrupts hydrogen bonding involving D334 thereby allowing the kinase to lock into the active conformation. Based on the simulations, three predictions were made: G83A was suggested to impede activation, K162M was suggested to perturb the interface between the N and C-domain leading to activation, and Q64C was hypothesized to stop folding of loop L16 thereby perturbing the homodimerization interface. Functional analysis of the mutants validated the predictions concerning the G83A and Q64C mutants. The K162M mutant did not autoactivate as predicted however, which may be due to the location of the residue on the protein surface near the ED substrate docking domain. PMID:21842857

  19. Importance of structural and chemical heterogeneity of activated carbon surfaces for adsorption of dibenzothiophene

    SciTech Connect

    Ania, C.O.; Bandosz, T.J.

    2005-08-16

    The performance of various activated carbons obtained from different carbon precursors (i.e., plastic waste, coal, and wood) as adsorbents for the desulfurization of liquid hydrocarbon fuels was evaluated. To increase surface heterogeneity, the carbon surface was modified by oxidation with ammonium persulfate. The results showed the importance of activated carbon pore sizes and surface chemistry for the adsorption of dibenzothiophene (DBT) from liquid phase. Adsorption of DBT on activated carbons is governed by two types of contributions: physical and chemical interactions. The former include dispersive interactions in the microporous network of the carbons. While the volume of micropores governs the amount physisorbed, mesopores control the kinetics of the process. On the other hand, introduction of surface functional groups enhances the performance of the activated carbons as a result of specific interactions between the acidic centers of the carbon and the basic structure of DBT molecule as well as sulfur-sulfur interactions.

  20. RNA Helicase Important for Listeria monocytogenes Hemolytic Activity and Virulence Factor Expression

    PubMed Central

    Netterling, Sakura; Bäreclev, Caroline; Vaitkevicius, Karolis

    2015-01-01

    RNA helicases have been shown to be important for the function of RNA molecules at several levels, although their putative involvement in microbial pathogenesis has remained elusive. We have previously shown that Listeria monocytogenes DExD-box RNA helicases are important for bacterial growth, motility, ribosomal maturation, and rRNA processing. We assessed the importance of the RNA helicase Lmo0866 (here named CshA) for expression of virulence traits. We observed a reduction in hemolytic activity in a strain lacking CshA compared to the wild type. This phenomenon was less evident in strains lacking other RNA helicases. The reduced hemolysis was accompanied by lower expression of major listerial virulence factors in the ΔcshA strain, mainly listeriolysin O, but also to some degree the actin polymerizing factor ActA. Reduced expression of these virulence factors in the strain lacking CshA did not, however, correlate with a decreased level of the virulence regulator PrfA. When combining the ΔcshA knockout with a mutation creating a constitutively active PrfA protein (PrfA*), the effect of the ΔcshA knockout on LLO expression was negated. These data suggest a role for the RNA helicase CshA in posttranslational activation of PrfA. Surprisingly, although the expression of several virulence factors was reduced, the ΔcshA strain did not demonstrate any reduced ability to infect nonphagocytic cells compared to the wild-type strain. PMID:26483402

  1. The Importance of Landscape Elements for Bat Activity and Species Richness in Agricultural Areas

    PubMed Central

    Heim, Olga; Treitler, Julia T.; Tschapka, Marco; Knörnschild, Mirjam; Jung, Kirsten

    2015-01-01

    Landscape heterogeneity is regarded as a key factor for maintaining biodiversity and ecosystem function in production landscapes. We investigated whether grassland sites at close vicinity to forested areas are more frequently used by bats. Considering that bats are important consumers of herbivorous insects, including agricultural pest, this is important for sustainable land management. Bat activity and species richness were assessed using repeated monitoring from May to September in 2010 with acoustic monitoring surveys on 50 grassland sites in the Biosphere Reserve Schorfheide-Chorin (North-East Germany). Using spatial analysis (GIS), we measured the closest distance of each grassland site to potentially connecting landscape elements (e.g., trees, linear vegetation, groves, running and standing water). In addition, we assessed the distance to and the percent land cover of forest remnants and urban areas in a 200 m buffer around the recording sites to address differences in the local landscape setting. Species richness and bat activity increased significantly with higher forest land cover in the 200 m buffer and at smaller distance to forested areas. Moreover, species richness increased in proximity to tree groves. Larger amount of forest land cover and smaller distance to forest also resulted in a higher activity of bats on grassland sites in the beginning of the year during May, June and July. Landscape elements near grassland sites also influenced species composition of bats and species richness of functional groups (open, edge and narrow space foragers). Our results highlight the importance of forested areas, and suggest that agricultural grasslands that are closer to forest remnants might be better buffered against outbreaks of agricultural pest insects due to higher species richness and higher bat activity. Furthermore, our data reveals that even for highly mobile species such as bats, a very dense network of connecting elements within the landscape is

  2. The Importance of Landscape Elements for Bat Activity and Species Richness in Agricultural Areas.

    PubMed

    Heim, Olga; Treitler, Julia T; Tschapka, Marco; Knörnschild, Mirjam; Jung, Kirsten

    2015-01-01

    Landscape heterogeneity is regarded as a key factor for maintaining biodiversity and ecosystem function in production landscapes. We investigated whether grassland sites at close vicinity to forested areas are more frequently used by bats. Considering that bats are important consumers of herbivorous insects, including agricultural pest, this is important for sustainable land management. Bat activity and species richness were assessed using repeated monitoring from May to September in 2010 with acoustic monitoring surveys on 50 grassland sites in the Biosphere Reserve Schorfheide-Chorin (North-East Germany). Using spatial analysis (GIS), we measured the closest distance of each grassland site to potentially connecting landscape elements (e.g., trees, linear vegetation, groves, running and standing water). In addition, we assessed the distance to and the percent land cover of forest remnants and urban areas in a 200 m buffer around the recording sites to address differences in the local landscape setting. Species richness and bat activity increased significantly with higher forest land cover in the 200 m buffer and at smaller distance to forested areas. Moreover, species richness increased in proximity to tree groves. Larger amount of forest land cover and smaller distance to forest also resulted in a higher activity of bats on grassland sites in the beginning of the year during May, June and July. Landscape elements near grassland sites also influenced species composition of bats and species richness of functional groups (open, edge and narrow space foragers). Our results highlight the importance of forested areas, and suggest that agricultural grasslands that are closer to forest remnants might be better buffered against outbreaks of agricultural pest insects due to higher species richness and higher bat activity. Furthermore, our data reveals that even for highly mobile species such as bats, a very dense network of connecting elements within the landscape is

  3. The Importance of Landscape Elements for Bat Activity and Species Richness in Agricultural Areas.

    PubMed

    Heim, Olga; Treitler, Julia T; Tschapka, Marco; Knörnschild, Mirjam; Jung, Kirsten

    2015-01-01

    Landscape heterogeneity is regarded as a key factor for maintaining biodiversity and ecosystem function in production landscapes. We investigated whether grassland sites at close vicinity to forested areas are more frequently used by bats. Considering that bats are important consumers of herbivorous insects, including agricultural pest, this is important for sustainable land management. Bat activity and species richness were assessed using repeated monitoring from May to September in 2010 with acoustic monitoring surveys on 50 grassland sites in the Biosphere Reserve Schorfheide-Chorin (North-East Germany). Using spatial analysis (GIS), we measured the closest distance of each grassland site to potentially connecting landscape elements (e.g., trees, linear vegetation, groves, running and standing water). In addition, we assessed the distance to and the percent land cover of forest remnants and urban areas in a 200 m buffer around the recording sites to address differences in the local landscape setting. Species richness and bat activity increased significantly with higher forest land cover in the 200 m buffer and at smaller distance to forested areas. Moreover, species richness increased in proximity to tree groves. Larger amount of forest land cover and smaller distance to forest also resulted in a higher activity of bats on grassland sites in the beginning of the year during May, June and July. Landscape elements near grassland sites also influenced species composition of bats and species richness of functional groups (open, edge and narrow space foragers). Our results highlight the importance of forested areas, and suggest that agricultural grasslands that are closer to forest remnants might be better buffered against outbreaks of agricultural pest insects due to higher species richness and higher bat activity. Furthermore, our data reveals that even for highly mobile species such as bats, a very dense network of connecting elements within the landscape is

  4. [Social support and physical activity in adolescents from public schools: the importance of family and friends].

    PubMed

    Prado, Crisley Vanessa; Lima, Alex Vieira; Fermino, Rogério César; Añez, Ciro Romelio Rodriguez; Reis, Rodrigo Siqueira

    2014-04-01

    The aim of this study was to verify the association between different types and sources of social support and physical activity among adolescents from Curitiba, Paraná State, Brazil. A school-based survey was conducted with a representative sample of adolescents from public schools (n = 1,469). Multiple regression models were used to test the association between weekly frequency and sources of social support from family and friends and weekly frequency of physical activity. Among boys, frequent company of family (PR: 2.88; 95%CI: 2.00-4.13) and friends (PR: 5.46; 95%CI: 2.33-12.78) and positive reinforcement from friends (PR: 1.81; 95%CI: 1.18-2.77) were positively associated with physical activity. Sporadic invitation by the family was negatively associated with physical activity (PR: 0.66; 95%CI: 0.46-1.14). For girls, frequent company of family (PR: 3.39; 95%CI: 1.49-7.69) and friends (PR: 4.06; 95%CI: 2.22-7.45) increased the likelihood of physical activity. Company of friends was the most important type of social support for physical activity among these adolescents.

  5. Halophilic Bacteria of Lunsu Produce an Array of Industrially Important Enzymes with Salt Tolerant Activity

    PubMed Central

    Gupta, Sonika; Sharma, Parul; Dev, Kamal; Sourirajan, Anuradha

    2016-01-01

    The halophilic bacterial isolates SS1, SS2, SS3, SS5, and SS8 were characterized for production of industrially important enzymes like amylase, protease, lipase, and glutaminase. Halophilic bacterial isolates SS1 and SS3 exhibited salt dependent extracellular amylase and protease activities. Both the halophilic isolates SS1 and SS3 exhibited maximum amylase and protease activities in the presence of 1.5 and 1.0 M NaCl, respectively, with the optimum pH 8 and temperature 40°C. SS2 showed maximum extracellular protease and lipase activities in the presence of 0.75 M NaCl, at optimum pH of 7, and temperature 37°C. The glutaminase activity of SS3 increased with increase in concentration of NaCl up to 2.5 M. The optimum pH and temperature for L-glutaminase activity of SS3 was 8 and 40°C, respectively. The combined hydrolytic activities of these halophilic bacterial isolates can be used for bioconversion of organic materials to useful products. PMID:26885394

  6. [Social support and physical activity in adolescents from public schools: the importance of family and friends].

    PubMed

    Prado, Crisley Vanessa; Lima, Alex Vieira; Fermino, Rogério César; Añez, Ciro Romelio Rodriguez; Reis, Rodrigo Siqueira

    2014-04-01

    The aim of this study was to verify the association between different types and sources of social support and physical activity among adolescents from Curitiba, Paraná State, Brazil. A school-based survey was conducted with a representative sample of adolescents from public schools (n = 1,469). Multiple regression models were used to test the association between weekly frequency and sources of social support from family and friends and weekly frequency of physical activity. Among boys, frequent company of family (PR: 2.88; 95%CI: 2.00-4.13) and friends (PR: 5.46; 95%CI: 2.33-12.78) and positive reinforcement from friends (PR: 1.81; 95%CI: 1.18-2.77) were positively associated with physical activity. Sporadic invitation by the family was negatively associated with physical activity (PR: 0.66; 95%CI: 0.46-1.14). For girls, frequent company of family (PR: 3.39; 95%CI: 1.49-7.69) and friends (PR: 4.06; 95%CI: 2.22-7.45) increased the likelihood of physical activity. Company of friends was the most important type of social support for physical activity among these adolescents. PMID:24896057

  7. How sotolon can impart a Madeira off-flavor to aged beers.

    PubMed

    Scholtes, Caroline; Nizet, Sabrina; Collin, Sonia

    2015-03-25

    4,5-Dimethyl-3-hydroxy-2(5H)-furanone or sotolon is known to impart powerful Madeira-oxidized-curry-walnut notes to various alcoholic beverages. It has been much studied in oxidized Jura flor-sherry wines, aged Roussillon sweet wines, and old Port wines, in which it contributes to the characteristic "Madeira-oxidized" aroma of these beverages. No scientific paper describes how sotolon might be involved in the Madeira off-flavor found in aged beers. The specific extraction procedure applied here allowed us to quantify this lactone in 7 special beers, at levels sometimes well above its threshold (from 5 to 42 μg/L after 6, 12, 18, and 24 months of natural aging, while unquantifiable in fresh beer). Investigation of spiked beers led us to highlight the key role of pro-oxidants and acetaldehyde. Addition of ascorbic acid without sulfites should be avoided by brewers, as the former would intensify sotolon synthesis. Acetoin, a beer fermentation byproduct, also emerged as possible precursor in beer when combined with serine. PMID:25727253

  8. A microRNA Imparts Robustness Against Environmental Fluctuation During Development

    PubMed Central

    Li, Xin; Cassidy, Justin J.; Reinke, Catherine A.; Fischboeck, Stephen; Carthew, Richard W.

    2009-01-01

    The microRNA miR-7 is perfectly conserved from annelids to humans, and yet some of the genes that it regulates in Drosophila are not regulated in mammals. We have explored the role of lineage restricted targets, using Drosophila, in order to better understand the evolutionary significance of microRNA-target relationships. From studies of two well characterized developmental regulatory networks, we find that miR-7 functions in several interlocking feedback and feedforward loops, and propose that its role in these networks is to buffer them against perturbation. To directly demonstrate this function for miR-7, we subjected the networks to temperature fluctuation and found that miR-7 is essential for the maintenance of regulatory stability under conditions of environmental flux. We suggest that some conserved microRNAs like miR-7 may enter into novel genetic relationships to buffer developmental programs against variation and impart robustness to diverse regulatory networks. PMID:19379693

  9. Importance of the redox state of cytochrome c during caspase activation in cytosolic extracts.

    PubMed Central

    Hampton, M B; Zhivotovsky, B; Slater, A F; Burgess, D H; Orrenius, S

    1998-01-01

    The export of cytochrome c from mitochondria to the cytoplasm has been detected during apoptosis. Addition of cytochrome c to cytosolic extracts can activate the caspases, suggesting that this export could be an important intracellular signal for initiating the apoptotic programme. We have investigated the mechanism of caspase activation by cytochrome c. Mitochondrial cytochrome c normally shuttles electrons between complexes III and IV of the electron transport chain. Interaction with these complexes is dependent on electrostatic interactions via a polylysine binding pocket. Cytosolic caspase activation was only observed with intact holocytochrome c, and increasing the ionic composition of the extracts prevented activation, suggesting that stringent allosteric interactions between cytochrome c and other cytoplasmic factors are necessary. Cytochrome c was fully reduced within 5 min of addition to the cytosolic extracts. Potassium ferricyanide could maintain cytochrome c in an oxidized state, but care was taken to use ferricyanide at concentrations where its polyanion effect did not cause interference. The oxidized form of cytochrome c was able to activate the caspases. We conclude that reduced cytochrome c will function in the cytoplasm; however, its reduction is not a critical step, and electron transfer from cytochrome c to its cytoplasmic-binding partner(s) is not necessary in the pathway leading to apoptosis. PMID:9405280

  10. Active transporters as enzymes: an energetic framework applied to major facilitator superfamily and ABC importer systems.

    PubMed

    Shilton, Brian H

    2015-04-15

    Active membrane transporters are dynamic molecular machines that catalyse transport across a membrane by coupling solute movement to a source of energy such as ATP or a secondary ion gradient. A central question for many active transporters concerns the mechanism by which transport is coupled to a source of energy. The transport process and associated energetic coupling involve conformational changes in the transporter. For efficient transport, the conformational changes must be tightly regulated and they must link energy use to movement of the substrate across the membrane. The present review discusses active transport using the well-established energetic framework for enzyme-mediated catalysis. In particular, membrane transport systems can be viewed as ensembles consisting of low-energy and high-energy conformations. The transport process involves binding interactions that selectively stabilize the higher energy conformations, and in this way promote conformational changes in the system that are coupled to decreases in free energy and substrate translocation. The major facilitator superfamily of secondary active transporters is used to illustrate these ideas, which are then be expanded to primary active transport mediated by ABC (ATP-binding cassette) import systems, with a focus on the well-studied maltose transporter.

  11. Isoleucine 61 is important for the hemolytic activity of pyolysin of Trueperella pyogenes.

    PubMed

    Yan, Minghui; Hu, Yunhao; Bao, Jun; Xiao, Ya; Zhang, Yue; Yang, Lingxiao; Wang, Junwei; Zhang, Wenlong

    2016-01-15

    Pyolysin (PLO) is a hemolysin secreted by Trueperella pyogenes (T. pyogenes) and is important for the pathogenicity of T. pyogenes. Oligomerization of PLO monomers is a critical step in the process of hemolysis. However, the mechanisms of intermolecular interaction of PLO monomers are still not clearly illuminated. In this study, two monoclonal antibodies (mAbs) against PLO, named AP-1A3 and AP-4F12, respectively, were generated firstly, of which AP-1A3 showed no or undetectable hemolysis inhibition activity against recombinant PLO (rPLO), whereas AP-4F12 could markedly inhibit the hemolytic activity of rPLO. Epitope mapping revealed that AP-1A3 recognized amino acid residues ranging from 64 to 79 of mature PLO (91-106 including the signal peptide), whereas AP-4F12 recognized amino acid residues ranging from 58 to 75 (85-102 including the signal peptide). Comparison of the amino acid sequence of two epitopes revealed that six amino acid residues ranging from 58 to 63 of PLO were associated with the hemolytic activity of PLO. Alanine scan showed that substitution of each amino acid ranging from 58 to 62 with alanine had apparent impact on the hemolytic activity of rPLO, especially for the substitution of isoleucine 61 which caused almost complete loss of hemolytic activity of rPLO. Our findings identified a region in PLO and an amino acid in that region might play important role in the process of oligomerization of PLO monomers. PMID:26711048

  12. PEGylated ofloxacin nanoparticles render strong antibacterial activity against many clinically important human pathogens.

    PubMed

    Marslin, Gregory; Revina, Ann Mary; Khandelwal, Vinoth Kumar Megraj; Balakumar, Krishnamoorthy; Sheeba, Caroline J; Franklin, Gregory

    2015-08-01

    The rise of bacterial resistance against important drugs threatens their clinical utility. Fluoroquinones, one of the most important classes of contemporary antibiotics has also reported to suffer bacterial resistance. Since the general mechanism of bacterial resistance against fluoroquinone antibiotics (e.g. ofloxacin) consists of target mutations resulting in reduced membrane permeability and increased efflux by the bacteria, strategies that could increase bacterial uptake and reduce efflux of the drug would provide effective treatment. In the present study, we have compared the efficiencies of ofloxacin delivered in the form of free drug (OFX) and as nanoparticles on bacterial uptake and antibacterial activity. Although both poly(lactic-co-glycolic acid) (OFX-PLGA) and methoxy poly(ethylene glycol)-b-poly(lactic-co-glycolic acid) (OFX-mPEG-PLGA) nanoformulations presented improved bacterial uptake and antibacterial activity against all the tested human bacterial pathogens, namely, Escherichia coli, Proteus vulgaris, Salmonella typhimurium, Pseudomonas aeruginosa, Klebsiella pneumoniae and Staphylococcus aureus, OFX-mPEG-PLGA showed significantly higher bacterial uptake and antibacterial activity compared to OFX-PLGA. We have also found that mPEG-PLGA nanoencapsulation could significantly inhibit Bacillus subtilis resistance development against OFX.

  13. Determination of Importance Evaluation for Exploratory Studies Facility (ESF) Subsurface Testing Activities

    SciTech Connect

    C.J. Byrne

    2001-02-20

    This Determination of Importance Evaluation (DIE) applies to the Subsurface Exploratory Studies Facility (ESF), encompassing the Topopah Spring (TS) Loop from Station 0+00 meters (m) at the North Portal to breakthrough at the South Portal (approximately 78+77 m), and ancillary test and operation support areas including the Enhanced Characterization of the Repository Block (ECRB) Cross Drift. This evaluation applies specifically to site characterization testing activities ongoing and planned in the Subsurface ESF. ESF site characterization activities are being performed to obtain the information necessary to determine whether the Yucca Mountain Site is suitable as a geologic repository for spent nuclear fuel and high-level radioactive waste. A more detailed description of these testing activities is provided in Section 6 of this DIE. Generally, the construction and operation of excavations associated with these testing activities are evaluated in the DIE for the Subsurface ESF (CRWMS M&O 1999a) and the DIE for the ESF ECRB Cross Drift (CRWMS M&O 2000a). The scope of this DIE also entails the proposed Unsaturated Zone (UZ) Transport Test at Busted Butte. Although, not a part of the TS Loop or ECRB Cross Drift, the associated testing activities are Subsurface testing activities. Busted Butte is located to the south south-east of the TS Loop and is outside the Conceptual Controlled Area Boundary (CCAB). These activities provide access to the Calico Hills (CH) geologic structure. In the case of Busted Butte, construction and operation of excavations are evaluated herein (since this activity was not previously evaluated in CRWMS M&O 1999a). The objectives of this DIE are to determine whether Subsurface ESF testing, and associated activities, could potentially impact site characterization testing and/or the waste isolation capabilities of the site. Controls needed to limit any potential impacts are identified in Section 13. The validity and veracity of the individual

  14. Genome sequence of Wickerhamomyces anomalus DSM 6766 reveals genetic basis of biotechnologically important antimicrobial activities.

    PubMed

    Schneider, Jessica; Rupp, Oliver; Trost, Eva; Jaenicke, Sebastian; Passoth, Volkmar; Goesmann, Alexander; Tauch, Andreas; Brinkrolf, Karina

    2012-05-01

    The ascomycetous yeast Wickerhamomyces anomalus (formerly Pichia anomala and Hansenula anomala) exhibits antimicrobial activities and flavoring features that are responsible for its frequent association with food, beverage and feed products. However, limited information on the genetic background of this yeast and its multiple capabilities are currently available. Here, we present the draft genome sequence of the neotype strain W. anomalus DSM 6766. On the basis of pyrosequencing, a de novo assembly of this strain resulted in a draft genome sequence with a total size of 25.47 Mbp. An automatic annotation using RAPYD generated 11 512 protein-coding sequences. This annotation provided the basis to analyse metabolic capabilities, phylogenetic relationships, as well as biotechnologically important features and yielded novel candidate genes of W. anomalus DSM 6766 coding for proteins participating in antimicrobial activities. PMID:22292503

  15. Determination of Importance Evaluation for Exploratory Studies Facility (ESF) Subsurface Testing Activities

    SciTech Connect

    S. Goodin

    2002-07-22

    This Determination of Importance Evaluation (DIE) applies to the Subsurface Exploratory Studies Facility (ESF), encompassing the Topopah Spring (TS) Loop from Station 0+00 meters (m) at the North Portal to breakthrough at the South Portal (approximately 78+77 m), and ancillary test and operation support areas including the Enhanced Characterization of the Repository Block (ECRB) Cross Drift. This evaluation applies specifically to site characterization testing activities ongoing and planned in the Subsurface ESF. ESF site characterization activities are being performed to obtain the information necessary to determine whether the Yucca Mountain Site is suitable as a geologic repository for spent nuclear fuel and high-level radioactive waste. A more detailed description of these testing activities is provided in Section 6 of this DIE. Generally, the construction and operation of excavations associated with these testing activities are evaluated in the DIE for the Subsurface ESF (CRWMS M&O 1999a) and the DIE for the ESF ECRB Cross Drift (CRWMS M&O 2000a). The scope of this DIE also entails the proposed Unsaturated Zone (UZ) Transport Test at Busted Butte. Although, not a part of the TS Loop or ECRB Cross Drift, the associated testing activities are Subsurface testing activities. Busted Butte is located to the south south-east of the TS Loop and is outside the Conceptual Controlled Area Boundary (CCAB). These activities provide access to the Calico Hills (CH) geologic structure. In the case of Busted Butte, construction and operation of excavations are evaluated herein (since this activity was not previously evaluated in CRWMS M&O 1999a). The objectives of this DIE are to determine whether Subsurface ESF testing, and associated activities, could potentially impact site characterization testing and/or the waste isolation capabilities of the site. Controls needed to limit any potential impacts are identified in Section 13. The validity and veracity of the individual

  16. The multidrug resistance protein 1: a functionally important activation marker for murine Th1 cells.

    PubMed

    Prechtl, S; Roellinghoff, M; Scheper, R; Cole, S P; Deeley, R G; Lohoff, M

    2000-01-15

    Previously, we described the expression of an energy-dependent pump in resting murine Th2 (but not resting Th1) cells which extruded the fluorescent dye Fluo-3. After stimulation with Ag and APCs, Th1 cells also expressed this pump. Furthermore, expression of the murine multidrug resistance protein 1 (mrp1) correlated with the presence of the pump. In this study, we report that Fluo-3 is indeed transported by murine mrp1 or its human ortholog MRP1, as revealed by transfection of HEK 293 cells with mrp1 or MRP1 cDNA. Like antigenic activation, IL-2 dose-dependently enhanced the Fluo-3-extruding activity in murine Th1 cells. Although TNF-alpha and IL-12 by themselves only weakly enhanced Fluo-3 extrusion, each of them did so in strong synergism with IL-2. An Ab directed against mrp1 was used to quantify the expression of mrp1 protein in T cells at the single-cell level. Like the Fluo-3 pump, mrp1 protein expression was enhanced by IL-2. Immunohistochemical studies using confocal laser microscopy indicated that mrp1 is localized mainly at the plasma membrane. In addition, protein expression of mrp1 was induced in Vbeta8+CD4+ T cells 12 h after in vivo application of Staphylococcal enterotoxin B. Finally, mrp1 was functionally relevant during the activation process of Th1 cells, because T cell activation could be suppressed by exposure of cells to the mrp1 inhibitor MK571. Thus, we present mrp1 as a novel, functionally important activation marker for Th1 cells and short-term in vivo activated CD4+ T cells, whereas its expression seems to be constitutive in Th2 cells.

  17. Activity and biological effects of neem products against arthropods of medical and veterinary importance.

    PubMed

    Mulla, M S; Su, T

    1999-06-01

    Botanical insecticides are relatively safe and degradable, and are readily available sources of biopesticides. The most prominent phytochemical pesticides in recent years are those derived from neem trees, which have been studied extensively in the fields of entomology and phytochemistry, and have uses for medicinal and cosmetic purposes. The neem products have been obtained from several species of neem trees in the family Meliaceae. Six species in this family have been the subject of botanical pesticide research. They are Azadirachta indica A. Juss, Azadirachta excelsa Jack, Azadirachta siamens Valeton, Melia azedarach L., Melia toosendan Sieb. and Zucc., and Melia volkensii Gürke. The Meliaceae, especially A. indica (Indian neem tree), contains at least 35 biologically active principles. Azadirachtin is the predominant insecticidal active ingredient in the seed, leaves, and other parts of the neem tree. Azadirachtin and other compounds in neem products exhibit various modes of action against insects such as antifeedancy, growth regulation, fecundity suppression and sterilization, oviposition repellency or attractancy, changes in biological fitness, and blocking development of vector-borne pathogens. Some of these bioactivity parameters of neem products have been investigated at least in some species of insects of medical and veterinary importance, such as mosquitoes, flies, triatomines, cockroaches, fleas, lice, and others. Here we review, synthesize, and analyze published information on the activity, modes of action, and other biological effects of neem products against arthropods of medical and veterinary importance. The amount of information on the activity, use, and application of neem products for the control of disease vectors and human and animal pests is limited. Additional research is needed to determine the potential usefulness of neem products in vector control programs. PMID:10412110

  18. A Meloidogyne incognita effector is imported into the nucleus and exhibits transcriptional activation activity in planta.

    PubMed

    Zhang, Lei; Davies, Laura J; Elling, Axel A

    2015-01-01

    Root-knot nematodes are sedentary biotrophic endoparasites that maintain a complex interaction with their host plants. Nematode effector proteins are synthesized in the oesophageal glands of nematodes and secreted into plant tissue through a needle-like stylet. Effectors characterized to date have been shown to mediate processes essential for nematode pathogenesis. To gain an insight into their site of action and putative function, the subcellular localization of 13 previously isolated Meloidogyne incognita effectors was determined. Translational fusions were created between effectors and EGFP-GUS (enhanced green fluorescent protein-β-glucuronidase) reporter genes, which were transiently expressed in tobacco leaf cells. The majority of effectors localized to the cytoplasm, with one effector, 7H08, imported into the nuclei of plant cells. Deletion analysis revealed that the nuclear localization of 7H08 was mediated by two novel independent nuclear localization domains. As a result of the nuclear localization of the effector, 7H08 was tested for the ability to activate gene transcription. 7H08 was found to activate the expression of reporter genes in both yeast and plant systems. This is the first report of a plant-parasitic nematode effector with transcriptional activation activity.

  19. Antimicrobial activity of some important Adiantum species used traditionally in indigenous systems of medicine.

    PubMed

    Singh, Meenakshi; Singh, Neha; Khare, P B; Rawat, A K S

    2008-01-17

    Adiantum Linn. of Adiantaceae family is one of the most common and widely distributed species. Ethnomedicinally, the genus is important and popularly known as "Hansraj" in Ayurvedic System of Medicine. It has been used in cold, tumors of spleen, liver and other viscera, skin diseases, bronchitis and inflammatory diseases. It is also considered as tonic and diuretic. In the present study its four important species, i.e. Adiantum capillus-veneris, Adiantum peruvianum, Adiantum venustum and Adiantum caudatum were collected and extracted with methanol. These extracts were tested for their antimicrobial agents against five gram positive, six gram negative (including multiresistant bacteria Staphylococcus aureus) and eight fungal strains using standard microdilution assay. The maximum activity was exhibited by the methanolic extract of Adiantum venustum followed by Adiantum capillus-veneris, Adiantum peruvianum and Adiantum caudatum. The methanolic extract of Adiantum capillus-veneris had very low MIC value (0.48 microg/ml) against Escherichia coli whereas, Adiantum venustum extract against Aspergillus terreus with MIC of 0.97 microg/ml. Total phenolic constituents of Adiantum species viz. Adiantum venustum, Adiantum capillus-veneris, Adiantum peruvianum and Adiantum caudatum were 0.81% (w/w), 0.83% (w/w), 0.71% (w/w) and 0.52% (w/w), respectively (as gallic acid equivalent); implying that the observed activity could be related to the amount of phenolics. PMID:17997240

  20. The nucleotide-binding domain of NLRC5 is critical for nuclear import and transactivation activity

    SciTech Connect

    Meissner, Torsten B.; Li, Amy; Liu, Yuen-Joyce; Gagnon, Etienne; Kobayashi, Koichi S.

    2012-02-24

    Highlights: Black-Right-Pointing-Pointer NLRC5 requires an intact NLS for its function as MHC class I transactivator. Black-Right-Pointing-Pointer Nuclear presence of NLRC5 is required for MHC class I induction. Black-Right-Pointing-Pointer Nucleotide-binding controls nuclear import and transactivation activity of NLRC5. -- Abstract: Major histocompatibility complex (MHC) class I and class II are crucial for the function of the human adaptive immune system. A member of the NLR (nucleotide-binding domain, leucine-rich repeat) protein family, NLRC5, has recently been identified as a transcriptional regulator of MHC class I and related genes. While a 'master regulator' of MHC class II genes, CIITA, has long been known, NLRC5 specifically associates with and transactivates the proximal promoters of MHC class I genes. In this study, we analyzed the molecular requirements of NLRC5 nuclear import and transactivation activity. We show that NLRC5-mediated MHC class I gene induction requires an intact nuclear localization signal and nuclear distribution of NLRC5. In addition, we find that the nucleotide-binding domain (NBD) of NLRC5 is critical not only for nuclear translocation but also for the transactivation of MHC class I genes. Changing the cellular localization of NLRC5 is likely to immediately impact MHC class I expression as well as MHC class I-mediated antigen presentation. NLRC5 may thus provide a promising target for the modulation of MHC class I antigen presentation, especially in the setting of transplant medicine.

  1. Importance of being Nernst: Synaptic activity and functional relevance in stem cell-derived neurons

    PubMed Central

    Bradford, Aaron B; McNutt, Patrick M

    2015-01-01

    Functional synaptogenesis and network emergence are signature endpoints of neurogenesis. These behaviors provide higher-order confirmation that biochemical and cellular processes necessary for neurotransmitter release, post-synaptic detection and network propagation of neuronal activity have been properly expressed and coordinated among cells. The development of synaptic neurotransmission can therefore be considered a defining property of neurons. Although dissociated primary neuron cultures readily form functioning synapses and network behaviors in vitro, continuously cultured neurogenic cell lines have historically failed to meet these criteria. Therefore, in vitro-derived neuron models that develop synaptic transmission are critically needed for a wide array of studies, including molecular neuroscience, developmental neurogenesis, disease research and neurotoxicology. Over the last decade, neurons derived from various stem cell lines have shown varying ability to develop into functionally mature neurons. In this review, we will discuss the neurogenic potential of various stem cells populations, addressing strengths and weaknesses of each, with particular attention to the emergence of functional behaviors. We will propose methods to functionally characterize new stem cell-derived neuron (SCN) platforms to improve their reliability as physiological relevant models. Finally, we will review how synaptically active SCNs can be applied to accelerate research in a variety of areas. Ultimately, emphasizing the critical importance of synaptic activity and network responses as a marker of neuronal maturation is anticipated to result in in vitro findings that better translate to efficacious clinical treatments. PMID:26240679

  2. Comparison of Antioxidant Activities of Hydrolysates of Domestic and Imported Skim Milk Powders Treated with Papain

    PubMed Central

    Ha, Go Eun; Chang, Oun Ki; Han, Gi Sung; Ham, Jun Sang; Park, Beom-Young; Jeong, Seok-Geun

    2015-01-01

    Milk proteins have many potential sequences within their primary structure, each with a specific biological activity. In this study, we compared and investigated the bioactivities of hydrolysates of the domestic (A, B) and imported (C, D) skim milk powders generated using papain digestion. MALDI-TOF analysis revealed that all milk powder proteins were intact, indicating no autolysis. Electrophoretic analysis of hydrolysates showed papain treatment caused degradation of milk proteins into peptides of various size. The antioxidant activity of the hydrolysates, determined using 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and total phenolic contents (TPC) assays, increased with incubation times. In all skim milk powders, the antioxidant activities of hydrolysates were highest following 24 h papain treatment (TPC: A, 196.48 μM GE/L; B, 194.52 μM GE/L; C, 194.76 μM GE/L; D, 163.75 μM GE/L; ABTS: A, 75%; B, 72%; C, 72%; D, 57%). The number of peptide derived from skim milk powders, as determined by LC-MS/MS, was 308 for A, 283 for B, 208 for C, and 135 for D. Hydrolysate A had the highest antioxidant activity and the most potential antioxidant peptides amongst the four skim milk powder hydrolysates. A total of 4 β-lactoglobulin, 4 αs1-casein, and 56 β-casein peptide fragments were identified as potential antioxidant peptides in hydrolysate A by LC-MS/MS. These results suggest that domestic skim milk could have applications in various industries, i.e., in the development of functional foods. PMID:26761850

  3. Increasing ocean temperatures reduce activity patterns of a large commercially important coral reef fish.

    PubMed

    Johansen, J L; Messmer, V; Coker, D J; Hoey, A S; Pratchett, M S

    2014-04-01

    Large-bodied fish are critical for sustaining coral reef fisheries, but little is known about the vulnerability of these fish to global warming. This study examined the effects of elevated temperatures on the movement and activity patterns of the common coral trout Plectropomus leopardus (Serranidae), which is an important fishery species in tropical Australia and throughout the Indo West-Pacific. Adult fish were collected from two locations on Australia's Great Barrier Reef (23°S and 14°S) and maintained at one of four temperatures (24, 27, 30, 33 °C). Following >4 weeks acclimation, the spontaneous swimming speeds and activity patterns of individuals were recorded over a period of 12 days. At 24-27 °C, spontaneous swimming speeds of common coral trout were 0.43-0.45 body lengths per second (bls(-1)), but dropped sharply to 0.29 bls(-1) at 30 °C and 0.25 bls(-1) at 33 °C. Concurrently, individuals spent 9.3-10.6% of their time resting motionless on the bottom at 24-27 °C, but this behaviour increased to 14.0% at 30 °C and 20.0% of the time at 33 °C (mean ± SE). The impact of temperature was greatest for smaller individuals (<45 cm TL), showing significant changes to swimming speeds across every temperature tested, while medium (45-55 cm TL) and large individuals (>55 cm TL) were first affected by 30 °C and 33 °C, respectively. Importantly, there was some indication that populations can adapt to elevated temperature if presented with adequate time, as the high-latitude population decreased significantly in swimming speeds at both 30 °C and 33 °C, while the low-latitude population only showed significant reductions at 33 °C. Given that movement and activity patterns of large mobile species are directly related to prey encounter rates, ability to capture prey and avoid predators, any reductions in activity patterns are likely to reduce overall foraging and energy intake, limit the energy available for growth and reproduction, and affect the fitness and

  4. Importance of neutron energy distribution in borehole activation analysis in relatively dry, low-porosity rocks

    USGS Publications Warehouse

    Senftle, F.E.; Moxham, R.M.; Tanner, A.B.; Philbin, P.W.; Boynton, G.R.; Wager, R.E.

    1977-01-01

    To evaluate the importance of variations in the neutron energy distribution in borehole activation analysis, capture gamma-ray measurements were made in relatively dry, low-porosity gabbro of the Duluth Complex. Although sections of over a meter of solid rock were encountered in the borehole, there was significant fracturing with interstitial water leading to a substantial variation of water with depth in the borehole. The linear-correlation coefficients calculated for the peak intensities of several elements compared to the chemical core analyses were generally poor throughout the depth investigated. The data suggest and arguments are given which indicate that the variation of the thermal-to-intermediate-to-fast neutron flux density as a function of borehole depth is a serious source of error and is a major cause of the changes observed in the capture gamma-ray peak intensities. These variations in neutron energy may also cause a shift in the observed capture gamma-ray energy.

  5. Hydrothermal activity at slow-spreading ridges: variability and importance of magmatic controls

    NASA Astrophysics Data System (ADS)

    Escartin, Javier

    2016-04-01

    less studied, similar hydrothermal systems are found elsewhere associated to other central volcanoes along the ridge axis (e.g., Menez Gwenn at the Mid-Atlantic Ridge and Soria Mornia or Troll Wall at the Arctic Ridges). Long-lived hydrothermal activity plays an important role in controlling the thermal structure of the lithosphere and its accretion at and near-axis, and also determining the distribution and biogeography of vent communities. Along slow-spreading segments, long-lived hydrothermal activity can be provided both by volcanic systems (e.g., Lucky Strike) and tectonic systems (oceanic detachment faults). While magmatic and hydrothermal activity is relatively well understood now in volcanic systems (e.g., Lucky Strike), tectonic systems (oceanic detachment faults) require further integrated studies to constrain the links between long-lived localization of deformation along oceanic detachment faults, hydrothermal activity, and origin and nature of off-axis heat sources animating hydrothermal circulation.

  6. Active nuclear import and export pathways regulate E2F-5 subcellular localization.

    PubMed

    Apostolova, Margarita D; Ivanova, Iordanka A; Dagnino, Carla; D'Souza, Sudhir J A; Dagnino, Lina

    2002-09-13

    Epidermal keratinocyte differentiation is accompanied by differential regulation of E2F genes, including up-regulation of E2F-5 and its concomitant association with the retinoblastoma family protein p130. This complex appears to play a role in irreversible withdrawal from the cell cycle in differentiating keratinocytes. We now report that keratinocyte differentiation is also accompanied by changes in E2F-5 subcellular localization, from the cytoplasm to the nucleus. To define the molecular determinants of E2F-5 nuclear import, we tested its ability to enter the nucleus in import assays in vitro using digitonin-permeabilized cells. We found that E2F-5 enters the nucleus through mediated transport processes that involve formation of nuclear pore complexes. It has been proposed that E2F-4 and E2F-5, which lack defined nuclear localization signal (NLS) consensus sequences, enter the nucleus in association with NLS-containing DP-2 or pRB family proteins. However, we show that nuclear import of E2F-5 only requires the first N-terminal 56 amino acid residues and is not dependent on interaction with DP or pRB family proteins. Because E2F-5 is predominantly cytoplasmic in undifferentiated keratinocytes and in other intact cells, we also examined whether this protein is subjected to active nuclear export. Indeed, E2F-5 is exported from the nucleus through leptomycin B-sensitive, CRM1-mediated transport, through a region corresponding to amino acid residues 130-154. This region excludes the DNA- and the p130-binding domains. Thus, the subcellular distribution of E2F-5 is tightly regulated in intact cells, through multiple functional domains that direct nucleocytoplasmic shuttling of this protein.

  7. Identification of a protein with antioxidant activity that is important for the protection against beer ageing.

    PubMed

    Wu, Ming J; Clarke, Frank M; Rogers, Peter J; Young, Paul; Sales, Narelle; O'Doherty, Patrick J; Higgins, Vincent J

    2011-01-01

    This study was carried out with fresh Australian lager beer which was sampled directly off the production line, the same samples aged for 12 weeks at 30 °C, and the vintage beer which was kept at 20 °C for 5 years. Characteristic Australian lager flavour was maintained in the fresh and vintage beers but was lost in the aged beer. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and free thiol group labelling analyses of beer proteins found that this flavour stability correlated with the presence of an unknown 10 kilodaltons (kDa) protein with a higher level of free thiols. The protein was purified by size-exclusion chromatography, then peptide sequencing and database matching identified it as the barley lipid transfer protein (LTP1). Further characterisation using diphenylpicrylhydrazyl (DPPH) free radical scavenging and a Saccharomyces cerevisiae-based antioxidant screening assay demonstrated that the LTP1 protein was active in DPPH reduction and antioxidant activity. The absence of free thiol in the aged beer indicates that the thiol functional groups within the LTP1 protein were saturated and suggests that it is important in the flavour stability of beer by maintaining reduction capacity during the ageing process.

  8. Identification of a Protein with Antioxidant Activity that is Important for the Protection against Beer Ageing

    PubMed Central

    Wu, Ming J.; Clarke, Frank M.; Rogers, Peter J.; Young, Paul; Sales, Narelle; O’Doherty, Patrick J.; Higgins, Vincent J.

    2011-01-01

    This study was carried out with fresh Australian lager beer which was sampled directly off the production line, the same samples aged for 12 weeks at 30 °C, and the vintage beer which was kept at 20 °C for 5 years. Characteristic Australian lager flavour was maintained in the fresh and vintage beers but was lost in the aged beer. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and free thiol group labelling analyses of beer proteins found that this flavour stability correlated with the presence of an unknown 10 kilodaltons (kDa) protein with a higher level of free thiols. The protein was purified by size-exclusion chromatography, then peptide sequencing and database matching identified it as the barley lipid transfer protein (LTP1). Further characterisation using diphenylpicrylhydrazyl (DPPH) free radical scavenging and a Saccharomyces cerevisiae-based antioxidant screening assay demonstrated that the LTP1 protein was active in DPPH reduction and antioxidant activity. The absence of free thiol in the aged beer indicates that the thiol functional groups within the LTP1 protein were saturated and suggests that it is important in the flavour stability of beer by maintaining reduction capacity during the ageing process. PMID:22016646

  9. Mineralization of naphtenic acids with thermally-activated persulfate: The important role of oxygen.

    PubMed

    Xu, Xiyan; Pliego, Gema; Zazo, Juan A; Casas, Jose A; Rodriguez, Juan J

    2016-11-15

    This study reports on the mineralization of model naphtenic acids (NAs) in aqueous solution by catalyst-free thermally-activated persulfate (PS) oxidation. These species are found to be pollutants in oil sands process-affected waters. The NAs tested include saturated-ring (cyclohexanecarboxylic and cyclohexanebutyric acids) and aromatic (2-naphthoic and 1,2,3,4-tetrahydro-2-naphthoic acids) structures, at 50mgL(-1)starting concentration. The effect of PS dose within a wide range (10-100% of the theoretical stoichiometric) and working temperature (40-97°C) was investigated. At 80°C and intitial pH=8 complete mineralization of the four NAs was achieved with 40-60% of the stoichiometric PS dose. This is explained because of the important contribution of oxygen, which was experimentally verified and was found to be more effective toward the NAs with a single cyclohexane ring than for the bicyclic aromatic-ring-bearing ones. The effect of chloride and bicarbonate was also checked. The former showed negative effect on the degradation rate of NAs whereas it was negligible or even positive for bicarbonate. The rate of mineralization was well described by simple pseudo-first order kinetics with values of the rate constants normalized to the PS dose within the range of 0.062-0.099h(-1). Apparent activation energy values between 93.7-105.3kJmol(-1) were obtained. PMID:27442986

  10. Determining importance and grading of items and activities for the Yucca Mountain Project

    SciTech Connect

    DeKlever, R.; Verna, B.

    1993-12-31

    Raytheon Services Nevada (RSN), in support of the Department of Energy`s (DOE) Yucca Mountain Project, has been responsible for the Title 2 designs of the initial structures, systems, and components for the Exploratory Studies Facility (ESF), and the creation of the design output documents for the Surface-Based Testing (SBT) programs. The ESF and SBT programs are major scientific contributors to the overall site characterization program which will determine the suitability of Yucca Mountain to contain a proposed High Level Nuclear Waste (HLNW) repository. Accurate, traceable and objective characterization and testing documentation that is germane to the protection of public health and safety, and the environment, and that satisfies all the requirements of 10 CFR Part 60(1), must be established, evaluated and accepted. To assure that these requirements are satisfied, specific design functions and products, including items and activities depicted within respective design output documents, are subjected to the requirements of an NRC and DOE-approved Quality Assurance (QA) program. An evaluation (classification) is applied to these items and activities to determine their importance to radiological safety (ITS) and waste isolation (ITWI). Subsequently, QA program controls are selected (grading) for the items and activities. RSN has developed a DOE-approved classification process that is based on probabilistic risk assessment (PRA) techniques and that uses accident/impact scenarios. Results from respective performance assessment and test interference evaluations are also integrated into the classification analyses for various items. The methodology and results of the RSN classification and grading processes, presented herein, relative to ESF and SBT design products, demonstrates a solid, defensible methodological basis for classification and grading.

  11. The functional importance of blood group-active molecules in human red blood cells.

    PubMed

    Anstee, D J

    2011-01-01

    Antigens of 23 of the 30 human blood group systems are defined by the amino acid sequence of red cell membrane proteins. The antigens of DI, RH, RHAG, MNS, GE and CO systems are carried on blood group-active proteins (Band 3, D and CE polypeptides, RhAG, Glycophorins A and B, Glycophorins C and D and Aquaporin 1, respectively) which are expressed at high levels (>200,000 copies/red cell). These major proteins contribute to essential red cell functions either directly as membrane transporters and by providing linkage to the underlying red cell skeleton or by facilitating the membrane assembly of the protein complexes involved in these processes. The proteins expressing antigens of the remaining 17 blood group systems are much less abundant (<20,000 copies/red cell) and their functional importance for the circulating red cell is largely unknown. Human gene knock-outs (null phenotypes) have been described for many of these minor blood group-active proteins, but only absence of Kx glycoprotein has been clearly linked with pathology directly related to the function of circulating red cells. Recent evidence suggesting the normal quality control system for glycoprotein synthesis is altered during the latter stages of red cell production raises the possibility that many of these low abundance blood group-active proteins are vestigial. In sickle cell disease and polycythaemia vera, elevated Lutheran glycoprotein expression may contribute to pathology. Dyserythropoiesis with reduced antigen expression can result from mutations in the erythroid transcription factors GATA-1 and EKLF.

  12. Resistance to Degradation and Cellular Distribution are Important Features for the Antitumor Activity of Gomesin

    PubMed Central

    Buri, Marcus V.; Domingues, Tatiana M.; Paredes-Gamero, Edgar J.; Casaes-Rodrigues, Rafael L.; Rodrigues, Elaine Guadelupe; Miranda, Antonio

    2013-01-01

    Many reports have shown that antimicrobial peptides exhibit anticancer abilities. Gomesin (Gm) exhibits potent cytotoxic activity against cancer cells by a membrane pore formation induced after well-orchestrated intracellular mechanisms. In this report, the replacements of the Cys by Ser or Thr, and the use D-amino acids in the Gm structure were done to investigate the importance of the resistance to degradation of the molecule with its cytotoxicity. [Thr2,6,11,15]-Gm, and [Ser2,6,11,15]-Gm exhibits low cytotoxicity, and low resistance to degradation, and after 24 h are present in localized area near to the membrane. Conversely, the use of D-amino acids in the analogue [D-Thr2,6,11,15]-D-Gm confers resistance to degradation, increases its potency, and maintained this peptide spread in the cytosol similarly to what happens with Gm. Replacements of Cys by Thr and Gln by L- or D-Pro ([D-Thr2,6,11,15, Pro9]-D-Gm, and [Thr2,6,11,15, D-Pro9]-Gm), which induced a similar β-hairpin conformation, also increase their resistance to degradation, and cytotoxicity, but after 24 h they are not present spread in the cytosol, exhibiting lower cytotoxicity in comparison to Gm. Additionally, chloroquine, a lysosomal enzyme inhibitor potentiated the effect of the peptides. Furthermore, the binding and internalization of peptides was determined, but a direct correlation among these factors was not observed. However, cholesterol ablation, which increase fluidity of cellular membrane, also increase cytotoxicity and internalization of peptides. β-hairpin spatial conformation, and intracellular localization/target, and the capability of entry are important properties of gomesin cytotoxicity. PMID:24312251

  13. Activated protein C resistance among postmenopausal women using transdermal estrogens: importance of progestogen

    PubMed Central

    Canonico, Marianne; Alhenc-Gelas, Martine; Plu-Bureau, Geneviève; Olié, Valérie; Scarabin, Pierre-Yves

    2010-01-01

    Introduction While the route of estrogen administration is known to be an important determinant of the thrombotic risk among postmenopausal women using hormone therapy, recent data have shown that norpregnane derivatives but not micronized progesterone would increase venous thromboembolism risk among transdermal estrogens users. However, differential effects of progesterone and norpregnanes on haemostasis have not yet been investigated. Methods We set up a cross-sectional study among healthy postmenopausal women aged 45 to 70 years. The impact of Activated Protein C (APC) on endogenous thrombin potential was investigated in plasma samples of 108 women who did not use any hormone therapy (n=40) or who were treated by transdermal estrogens combined with micronized progesterone (n=30) or norpregnane derivatives (n=38). Results After exclusion of women with factor V Leiden and/or G20210A prothrombin gene mutations, there was no significant change in APC sensitivity among women who used transdermal estrogens combined with micronized progesterone compared to non-users. Women using transdermal estrogens combined with norpregnanes were less sensitive to APC than were non-users (p=0.003) or users of transdermal estrogens combined with micronized progesterone (p=0.004). In addition, prothrombin fragment 1+2 concentration was higher in users of transdermal estrogens plus norpregnanes than in non-users (p=0.004). Other haemostatic parameters did not vary significantly across the different subgroups. Conclusion Transdermal estrogens combined with norpregnanes may induce an APC resistance and activate blood coagulation. These results provide a biological support to epidemiological data regarding the potential thrombogenic effects of norpregnanes. However, these findings need to be confirmed in a randomized trial. PMID:20613675

  14. Calcitriol imparts neuroprotection in vitro to midbrain dopaminergic neurons by upregulating GDNF expression.

    PubMed

    Orme, Rowan P; Bhangal, Manminder S; Fricker, Rosemary A

    2013-01-01

    During development a tightly controlled signaling cascade dictates the differentiation, maturation and survival of developing neurons. Understanding this signaling mechanism is important for developing therapies for neurodegenerative illnesses. In previous work we have sought to understand the complex signaling pathways responsible for the development of midbrain dopamine neurons using a proteomic approach. One protein we have identified as being expressed in developing midbrain tissue is the vitamin D receptor. Therefore we investigated the effect of the biologically active vitamin D3 metabolite, calcitriol, on primary fetal ventral mesencephalic cultures of dopamine neurons. We observed a dose responsive increase in numbers of rat primary dopamine neurons when calcitriol was added to culture media. Western blot data showed that calcitriol upregulated the expression of glial derived neurotrophic factor (GDNF). Blocking GDNF signaling could prevent calcitriol's ability to increase numbers of dopamine neurons. An apoptosis assay and cell birth dating experiment revealed that calcitriol increases the number of dopamine neurons through neuroprotection and not increased differentiation. This could have implications for future neuroprotective PD therapies.

  15. Calcitriol Imparts Neuroprotection In Vitro to Midbrain Dopaminergic Neurons by Upregulating GDNF Expression

    PubMed Central

    Orme, Rowan P.; Bhangal, Manminder S.; Fricker, Rosemary A.

    2013-01-01

    During development a tightly controlled signaling cascade dictates the differentiation, maturation and survival of developing neurons. Understanding this signaling mechanism is important for developing therapies for neurodegenerative illnesses. In previous work we have sought to understand the complex signaling pathways responsible for the development of midbrain dopamine neurons using a proteomic approach. One protein we have identified as being expressed in developing midbrain tissue is the vitamin D receptor. Therefore we investigated the effect of the biologically active vitamin D3 metabolite, calcitriol, on primary fetal ventral mesencephalic cultures of dopamine neurons. We observed a dose responsive increase in numbers of rat primary dopamine neurons when calcitriol was added to culture media. Western blot data showed that calcitriol upregulated the expression of glial derived neurotrophic factor (GDNF). Blocking GDNF signaling could prevent calcitriol’s ability to increase numbers of dopamine neurons. An apoptosis assay and cell birth dating experiment revealed that calcitriol increases the number of dopamine neurons through neuroprotection and not increased differentiation. This could have implications for future neuroprotective PD therapies. PMID:23626767

  16. Test-Retest Reliability and descriptive analyses of the Modified Important People and Activities (MIPA) Interview

    PubMed Central

    Zywiak, William H.; Stout, Robert L.; Braciszewski, Jordan M.; Wray, Tyler B.; Longabaugh, Richard

    2016-01-01

    Background The Important People and Activities (IPA) instrument assesses network characteristics and social support for drinking and abstinence. The IPA has garnered widespread use in the alcohol treatment field. We modified the IPA to assess HIV status, drug of choice, and IV drug use among social network members. Further, we queried frequency of unprotected sex, between the participant and network members. Aim Since this measure was modified, and the test-retest reliability of the IPA has only rarely been examined, we conducted a small substudy (n=26) to examine 1-week test-retest reliability of this measure. Methods Participants were individuals in a day treatment program with an SUD and/or AUD diagnosis. Results Drug of choice for the participants represented roughly equal thirds of heroin, cocaine, and alcohol. The sample was 62% female and 39% Latino/a. At pretest 198 persons were named on the MIPA (M=7.6 network members per subject). It was determined that 152 of the people were overlapping between the test and retest. Conclusions Percent agreement, ICCs, and kappas for the items ranged from acceptable to excellent across the two time periods. Classification of network members as positive, negative, or neutral influences on sobriety also demonstrated good to excellent kappas. PMID:27429606

  17. The July 2009 flurry of 3≤M<6 seismicity across Taiwan, and its possible relationship to coseismic and postseismic stresses imparted by the 1999 M=7.6 Chi-Chi earthquake

    NASA Astrophysics Data System (ADS)

    Stein, R. S.; Chan, C.

    2009-12-01

    An unusually productive sequence of twelve 3≤M<6 earthquakes occurred in central and northeastern Taiwan during 2-28 July 2009. These events have strike-slip and thrust focal mechanisms. We study the progression of these shocks, as well as smaller (M≥2) shocks since the time of the 21 September 1999 M=7.6 Chi-Chi event, and calculate of the impact of stress transferred by Chi-Chi on their occurrence. We estimate the Coulomb stress imparted by the coseismic slip, afterslip and viscoelastic deformation, all of which are important for this ramp-thrust event. Eleven out of the 12 July 2009 shocks sustained a Coulomb stress increase on at least one of their nodal planes caused by the coseismic and postseismic stress imparted by the Chi-Chi event (assuming a friction coefficient of 0.4). The stress increases range from 0.05 to 2.1 bars. Nevertheless, only 7 out of the 12 shocks can be associated with postseismic stress increases resolved onto their nodal planes, so the influence of postseismic stress remains equivocal. In examining the much more abundant M≥2 shocks, we find a good spatial correlation between the seismicity rate change and the calculated postseismic stress imparted 15 months after Chi-Chi. Here, the seismicity rate 15-75 months after the 1999 quake is divided by the rate during the 60 months before the 1999 quake. A very high rate of seismicity is observed just northeast of the northern end of the Chelungpu fault relative to the period immediately after the Chi-Chi shock. This pattern of seismicity appears to be associated with stress imparted by afterslip and viscoelastic deformation. Thus, the recent flurry of earthquakes seems largely consistent with the calculated evolution of stress and seismicity during the past decade, as does the occurrence of smaller events.

  18. Mg2+ imparts NMDA receptor subtype selectivity to the Alzheimer's drug memantine.

    PubMed

    Kotermanski, Shawn E; Johnson, Jon W

    2009-03-01

    N-methyl-D-aspartate receptors (NMDARs) mediate interneuronal communication and are broadly involved in nervous system physiology and pathology (Dingledine et al., 1999). Memantine, a drug that blocks the ion channel formed by NMDARs, is a widely prescribed treatment of Alzheimer's disease (Schmitt, 2005; Lipton, 2006; Parsons et al., 2007). Research on memantine's mechanism of action has focused on the NMDAR subtypes most highly expressed in adult cerebral cortex, NR1/2A and NR1/2B receptors (Cull-Candy and Leszkiewicz, 2004), and has largely ignored interactions with extracellular Mg(2+) (Mg(2+)(o)). Mg(2+)(o) is an endogenous NMDAR channel blocker that binds near memantine's binding site (Kashiwagi et al., 2002; Chen and Lipton, 2005). We report that a physiological concentration (1 mM) of Mg(2+)(o) decreased memantine inhibition of NR1/2A and NR1/2B receptors nearly 20-fold at a membrane voltage near rest. In contrast, memantine inhibition of the other principal NMDAR subtypes, NR1/2C and NR1/2D receptors, was decreased only approximately 3-fold. As a result, therapeutic memantine concentrations should have negligible effects on NR1/2A or NR1/2B receptor activity but pronounced effects on NR1/2C and NR1/2D receptors. Quantitative modeling showed that the voltage dependence of memantine inhibition also is altered by 1 mM Mg(2+)(o). We report similar results with the NMDAR channel blocker ketamine, a drug used to model schizophrenia (Krystal et al., 2003). These results suggest that currently hypothesized mechanisms of memantine and ketamine action should be reconsidered and that NR1/2C and/or NR1/2D receptors play a more important role in cortical physiology and pathology than previously appreciated.

  19. Escherichia coli Pyruvate Dehydrogenase Complex Is an Important Component of CXCL10-Mediated Antimicrobial Activity

    PubMed Central

    Schutte, Kirsten M.; Fisher, Debra J.; Burdick, Marie D.; Mehrad, Borna; Mathers, Amy J.; Mann, Barbara J.; Nakamoto, Robert K.

    2015-01-01

    Chemokines are best recognized for their role within the innate immune system as chemotactic cytokines, signaling and recruiting host immune cells to sites of infection. Certain chemokines, such as CXCL10, have been found to play an additional role in innate immunity, mediating CXCR3-independent killing of a diverse array of pathogenic microorganisms. While this is still not clearly understood, elucidating the mechanisms underlying chemokine-mediated antimicrobial activity may facilitate the development of novel therapeutic strategies effective against antibiotic-resistant Gram-negative pathogens. Here, we show that CXCL10 exerts antibacterial effects on clinical and laboratory strains of Escherichia coli and report that disruption of pyruvate dehydrogenase complex (PDHc), which converts pyruvate to acetyl coenzyme A, enables E. coli to resist these antimicrobial effects. Through generation and screening of a transposon mutant library, we identified two mutants with increased resistance to CXCL10, both with unique disruptions of the gene encoding the E1 subunit of PDHc, aceE. Resistance to CXCL10 also occurred following deletion of either aceF or lpdA, genes that encode the remaining two subunits of PDHc. Although PDHc resides within the bacterial cytosol, electron microscopy revealed localization of immunogold-labeled CXCL10 to the bacterial cell surface in both the E. coli parent and aceE deletion mutant strains. Taken together, our findings suggest that while CXCL10 interacts with an as-yet-unidentified component on the cell surface, PDHc is an important mediator of killing by CXCL10. To our knowledge, this is the first description of PDHc as a key bacterial component involved in the antibacterial effect of a chemokine. PMID:26553462

  20. Escherichia coli Pyruvate Dehydrogenase Complex Is an Important Component of CXCL10-Mediated Antimicrobial Activity.

    PubMed

    Schutte, Kirsten M; Fisher, Debra J; Burdick, Marie D; Mehrad, Borna; Mathers, Amy J; Mann, Barbara J; Nakamoto, Robert K; Hughes, Molly A

    2016-01-01

    Chemokines are best recognized for their role within the innate immune system as chemotactic cytokines, signaling and recruiting host immune cells to sites of infection. Certain chemokines, such as CXCL10, have been found to play an additional role in innate immunity, mediating CXCR3-independent killing of a diverse array of pathogenic microorganisms. While this is still not clearly understood, elucidating the mechanisms underlying chemokine-mediated antimicrobial activity may facilitate the development of novel therapeutic strategies effective against antibiotic-resistant Gram-negative pathogens. Here, we show that CXCL10 exerts antibacterial effects on clinical and laboratory strains of Escherichia coli and report that disruption of pyruvate dehydrogenase complex (PDHc), which converts pyruvate to acetyl coenzyme A, enables E. coli to resist these antimicrobial effects. Through generation and screening of a transposon mutant library, we identified two mutants with increased resistance to CXCL10, both with unique disruptions of the gene encoding the E1 subunit of PDHc, aceE. Resistance to CXCL10 also occurred following deletion of either aceF or lpdA, genes that encode the remaining two subunits of PDHc. Although PDHc resides within the bacterial cytosol, electron microscopy revealed localization of immunogold-labeled CXCL10 to the bacterial cell surface in both the E. coli parent and aceE deletion mutant strains. Taken together, our findings suggest that while CXCL10 interacts with an as-yet-unidentified component on the cell surface, PDHc is an important mediator of killing by CXCL10. To our knowledge, this is the first description of PDHc as a key bacterial component involved in the antibacterial effect of a chemokine.

  1. GlpC gene is responsible for biofilm formation and defense against phagocytes and imparts tolerance to pH and organic solvents in Proteus vulgaris.

    PubMed

    Wu, Y L; Liu, K S; Yin, X T; Fei, R M

    2015-01-01

    Biofilm-forming bacteria are highly resistant to antibiotics, host immune defenses, and other external conditions. The formation of biofilms plays a key role in colonization and infection. To explore the mechanism of biofilm formation, mutant strains of Proteus vulgaris XC 2 were generated by Tn5 random transposon insertion. Only one biofilm defective bacterial species was identified from among 500 mutants. Inactivation of the glpC gene coding an anaerobic glycerol-3-phosphate dehydrogenase subunit C was identified by sequence analysis of the biofilm defective strain. Differences were detected in the growth phenotypes of the wild-type and mutant strains under pH, antibiotic, and organic solvent stress conditions. Furthermore, we observed an increase in the phagocytosis of the biofilm defective strain by the mouse macrophage RAW264.7 cell line compared to the wild-type strain. This study shows that the glpC gene plays an important role in biofilm formation, in addition to imparting pH, organic solvent, and antibiotic tolerance, and defense against phagocytosis to Proteus sp. The results further clarified the mechanism of biofilm formation at the genomic level, and indicated the importance of the glpC gene in this process. This data may provide innovative therapeutic measures against P. vulgaris infections; furthermore, as an important crocodile pathogen, this study also has important significance in the protection of Chinese alligators. PMID:26400293

  2. GlpC gene is responsible for biofilm formation and defense against phagocytes and imparts tolerance to pH and organic solvents in Proteus vulgaris.

    PubMed

    Wu, Y L; Liu, K S; Yin, X T; Fei, R M

    2015-09-09

    Biofilm-forming bacteria are highly resistant to antibiotics, host immune defenses, and other external conditions. The formation of biofilms plays a key role in colonization and infection. To explore the mechanism of biofilm formation, mutant strains of Proteus vulgaris XC 2 were generated by Tn5 random transposon insertion. Only one biofilm defective bacterial species was identified from among 500 mutants. Inactivation of the glpC gene coding an anaerobic glycerol-3-phosphate dehydrogenase subunit C was identified by sequence analysis of the biofilm defective strain. Differences were detected in the growth phenotypes of the wild-type and mutant strains under pH, antibiotic, and organic solvent stress conditions. Furthermore, we observed an increase in the phagocytosis of the biofilm defective strain by the mouse macrophage RAW264.7 cell line compared to the wild-type strain. This study shows that the glpC gene plays an important role in biofilm formation, in addition to imparting pH, organic solvent, and antibiotic tolerance, and defense against phagocytosis to Proteus sp. The results further clarified the mechanism of biofilm formation at the genomic level, and indicated the importance of the glpC gene in this process. This data may provide innovative therapeutic measures against P. vulgaris infections; furthermore, as an important crocodile pathogen, this study also has important significance in the protection of Chinese alligators.

  3. Sound Perception: Rhythmic Brain Activity Really Is Important for Auditory Segregation.

    PubMed

    Snyder, Joel S

    2015-12-21

    A new study suggests that rhythmic brain activity plays a causal role in the perceptual segregation of sound patterns, rather than such activity simply being a non-functional by-product of sensory processing.

  4. Riverine Particulate Organic Carbon From a Pristine, Active Mountain Belt: The Importance of Vegetation and Landslides

    NASA Astrophysics Data System (ADS)

    Hilton, R. G.; Hovius, N.; Galy, A.

    2005-12-01

    The erosion and transfer of terrestrial organic carbon is an important component in the carbon cycle. The sequestration of this material in sedimentary basins may influence global climate. Mountain rivers play a crucial role in the routing of Particulate Organic Carbon (POC) because they have high sediment production rates, small storage potential and tendency to discharge sediment to the oceans at very high concentrations. We present new constraints on the sourcing and transfer of POC in the tectonically active Southern Alps of New Zealand, where anthropogenic disturbance is minimal. Riverine POC can be derived from bedrock (in the form of sedimentary kerogen), standing biomass and soils. In active mountain belts, landsliding dominates sediment production and transfer of POC from hillslopes to river channels. Landslide debris fans, river suspended load and bedload, and bedrock were sampled in catchments draining the rapidly eroding western Southern Alps, in order to assess the sourcing and transfer of POC. Samples were analyzed for percent organic carbon (Corg), percent nitrogen (N_ {org) and δ13C. Using C/N and δ13C as source proxies, we have determined the main sources of the POC. Landslide debris has Corg ~ 0.15% to 2.77%. The sand, silt and clay fractions of this material all have C/N ~ 10 to 40 and δ13C ~ -20‰ to -28‰. This indicates that POC in landslide debris is dominated by soil carbon (C/N ~ 16 to 25) mixed with hillslope vegetation (C/N ~ 40+) and diluted by bedrock (Corg = 0.01%, C/N = 4.7). Variations in the vegetation type of the pre-slide hillslopes and the depth of failure (i.e. volume of bedrock) give rise to variability of carbon concentration and source signature between landslides. In river suspended load, Corg ~ 0.5%, C/N ~ 15.6 and δ13C ~ - 23.5‰ at mean flow. These values are relatively uniform along the mountain belt. C_ {org} decreases with increasing suspended sediment concentration. We conclude that the bulk of the suspended

  5. In vitro activity of premafloxacin, a new extended-spectrum fluoroquinolone, against pathogens of veterinary importance.

    PubMed Central

    Watts, J L; Salmon, S A; Sanchez, M S; Yancey, R J

    1997-01-01

    The in vitro activity of premafloxacin against 673 veterinary pathogens was evaluated. Premafloxacin was equivalent to ciprofloxacin, enrofloxacin, and danofloxacin in activity against the gram-negative bacilli but was much more active (MIC for 90% of the strains tested [MIC90], 0.015 to 0.25 microg/ml) than the comparison antimicrobial agents (MIC90, 0.13 to 16.0 microg/ml) against the staphylococci, streptococci, and anaerobes tested. PMID:9145899

  6. Esterase in imported fire ants, Solenopsis invicata and S. richteri (Hymenoptera: Formicidae): activity, kinetics and variation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Black imported fire ant, Solenopsis richteri, is closely related to the notorious red imported fire ant, Solenopsis invicta. Despite being very similar in biology and behavior, S. invicta is a much more successful invader. In contrast to S. invicta that has invaded numberous countries and regions,...

  7. MAP kinases with distinct inhibitory functions impart signaling specificity during yeast differentiation.

    PubMed

    Madhani, H D; Styles, C A; Fink, G R

    1997-11-28

    Filamentous invasive growth of S. cerevisiae requires multiple elements of the mitogen-activated protein kinase (MAPK) signaling cascade that are also components of the mating pheromone response pathway. Here we show that, despite sharing several constituents, the two pathways use different MAP kinases. The Fus3 MAPK regulates mating, whereas the Kss1 MAPK regulates filamentation and invasion. Remarkably, in addition to their kinase-dependent activation functions, Kss1 and Fus3 each have a distinct kinase-independent inhibitory function. Kss1 inhibits the filamentation pathway by interacting with its target transcription factor Ste12. Fus3 has a different inhibitory activity that prevents the inappropriate activation of invasion by the pheromone response pathway. In the absence of Fus3, there is erroneous crosstalk in which mating pheromone now activates filamentation-specific gene expression using the Kss1 MAPK. PMID:9393860

  8. The 4'-hydroxyl group of resveratrol is functionally important for direct activation of PPARα.

    PubMed

    Takizawa, Yoshie; Nakata, Rieko; Fukuhara, Kiyoshi; Yamashita, Hiroshi; Kubodera, Hideo; Inoue, Hiroyasu

    2015-01-01

    Long-term moderate consumption of red wine is associated with a reduced risk of developing lifestyle-related diseases such as cardiovascular disease and cancer. Therefore, resveratrol, a constituent of grapes and various other plants, has attracted substantial interest. This study focused on one molecular target of resveratrol, the peroxisome proliferator activated receptor α (PPARα). Our previous study in mice showed that resveratrol-mediated protection of the brain against stroke requires activation of PPARα; however, the molecular mechanisms involved in this process remain unknown. Here, we evaluated the chemical basis of the resveratrol-mediated activation of PPARα by performing a docking mode simulation and examining the structure-activity relationships of various polyphenols. The results of experiments using the crystal structure of the PPARα ligand-binding domain and an analysis of the activation of PPARα by a resveratrol analog 4-phenylazophenol (4-PAP) in vivo indicate that the 4'-hydroxyl group of resveratrol is critical for the direct activation of PPARα. Activation of PPARα by 5 μM resveratrol was enhanced by rolipram, an inhibitor of phosphodiesterase (PDE) and forskolin, an activator of adenylate cyclase. We also found that resveratrol has a higher PDE inhibitory activity (IC50 = 19 μM) than resveratrol analogs trans-4-hydroxystilbene and 4-PAP (IC50 = 27-28 μM), both of which has only 4'-hydroxyl group, indicating that this 4'-hydroxyl group of resveratrol is not sufficient for the inhibition of PDE. This result is consistent with that 10 μM resveratrol has a higher agonistic activity of PPARα than these analogs, suggesting that there is a feedforward activation loop of PPARα by resveratrol, which may be involved in the long-term effects of resveratrol in vivo. PMID:25798826

  9. 75 FR 79342 - Taking and Importing Marine Mammals; Navy Training Activities Conducted Within the Northwest...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-12-20

    ... preamble to the final rule (75 FR 69296, November 10, 2010), that the taking resulting from the activities... findings, described in the preamble to the final rule (75 FR 45527, August 3, 2010), that the taking... Training Activities Conducted Within the Northwest Training Range Complex (NWTRC) and Military...

  10. IMPORTANCE OF ACTIVATED CARBON'S OXYGEN SURFACE FUNCTIONAL GROUPS ON ELEMENTAL MERCURY ADSORPTION

    EPA Science Inventory

    The effect of varying physical and chemical properties of activated carbons on adsorption of elemental mercury [Hg(0)] was studied by treating two activated carbons to modify their surface functional groups and pore structures. Heat treatment (1200 K) in nitrogen (N2), air oxidat...

  11. Functional Mapping of Protein Kinase A Reveals Its Importance in Adult Schistosoma mansoni Motor Activity

    PubMed Central

    de Saram, Paulu S. R.; Ressurreição, Margarida; Davies, Angela J.; Rollinson, David; Emery, Aidan M.; Walker, Anthony J.

    2013-01-01

    Cyclic AMP (cAMP)-dependent protein kinase/protein kinase A (PKA) is the major transducer of cAMP signalling in eukaryotic cells. Here, using laser scanning confocal microscopy and ‘smart’ anti-phospho PKA antibodies that exclusively detect activated PKA, we provide a detailed in situ analysis of PKA signalling in intact adult Schistosoma mansoni, a causative agent of debilitating human intestinal schistosomiasis. In both adult male and female worms, activated PKA was consistently found associated with the tegument, oral and ventral suckers, oesophagus and somatic musculature. In addition, the seminal vesicle and gynaecophoric canal muscles of the male displayed activated PKA whereas in female worms activated PKA localized to the ootype wall, the ovary, and the uterus particularly around eggs during expulsion. Exposure of live worms to the PKA activator forskolin (50 µM) resulted in striking PKA activation in the central and peripheral nervous system including at nerve endings at/near the tegument surface. Such neuronal PKA activation was also observed without forskolin treatment, but only in a single batch of worms. In addition, PKA activation within the central and peripheral nervous systems visibly increased within 15 min of worm-pair separation when compared to that observed in closely coupled worm pairs. Finally, exposure of adult worms to forskolin induced hyperkinesias in a time and dose dependent manner with 100 µM forskolin significantly increasing the frequency of gross worm movements to 5.3 times that of control worms (P≤0.001). Collectively these data are consistent with PKA playing a central part in motor activity and neuronal communication, and possibly interplay between these two systems in S. mansoni. This study, the first to localize a protein kinase when exclusively in an activated state in adult S. mansoni, provides valuable insight into the intricacies of functional protein kinase signalling in the context of whole schistosome physiology

  12. Functional mapping of protein kinase A reveals its importance in adult Schistosoma mansoni motor activity.

    PubMed

    de Saram, Paulu S R; Ressurreição, Margarida; Davies, Angela J; Rollinson, David; Emery, Aidan M; Walker, Anthony J

    2013-01-01

    Cyclic AMP (cAMP)-dependent protein kinase/protein kinase A (PKA) is the major transducer of cAMP signalling in eukaryotic cells. Here, using laser scanning confocal microscopy and 'smart' anti-phospho PKA antibodies that exclusively detect activated PKA, we provide a detailed in situ analysis of PKA signalling in intact adult Schistosoma mansoni, a causative agent of debilitating human intestinal schistosomiasis. In both adult male and female worms, activated PKA was consistently found associated with the tegument, oral and ventral suckers, oesophagus and somatic musculature. In addition, the seminal vesicle and gynaecophoric canal muscles of the male displayed activated PKA whereas in female worms activated PKA localized to the ootype wall, the ovary, and the uterus particularly around eggs during expulsion. Exposure of live worms to the PKA activator forskolin (50 µM) resulted in striking PKA activation in the central and peripheral nervous system including at nerve endings at/near the tegument surface. Such neuronal PKA activation was also observed without forskolin treatment, but only in a single batch of worms. In addition, PKA activation within the central and peripheral nervous systems visibly increased within 15 min of worm-pair separation when compared to that observed in closely coupled worm pairs. Finally, exposure of adult worms to forskolin induced hyperkinesias in a time and dose dependent manner with 100 µM forskolin significantly increasing the frequency of gross worm movements to 5.3 times that of control worms (P≤0.001). Collectively these data are consistent with PKA playing a central part in motor activity and neuronal communication, and possibly interplay between these two systems in S. mansoni. This study, the first to localize a protein kinase when exclusively in an activated state in adult S. mansoni, provides valuable insight into the intricacies of functional protein kinase signalling in the context of whole schistosome physiology.

  13. 31 CFR 560.505 - Importation of certain Iranian-origin services authorized; activities related to certain visa...

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... services authorized; activities related to certain visa categories authorized. 560.505 Section 560.505...; activities related to certain visa categories authorized. (a) The importation of Iranian-origin services into... with that purpose. (b) Persons otherwise qualified for a non-immigrant visa under categories A-3 and...

  14. 78 FR 36580 - Agency Information Collection Activities; Proposed Collection; Comments Requested: Import/Export...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-06-18

    ... previously published in the Federal Register at Volume 78 FR 19312, March 29, 2013, allowing for a 60 day... chemicals ephedrine, pseudoephedrine, and phenylpropanolamine, importers must report all information...

  15. In vitro antifungal activity of pneumocandin L-743,872 against a variety of clinically important molds.

    PubMed Central

    Del Poeta, M; Schell, W A; Perfect, J R

    1997-01-01

    The in vitro activity of the new antifungal drug pneumocandin L-743,872 against 55 isolates of clinically important molds was examined by an adapted macrobroth dilution method for yeasts. Pneumocandin L-743,872 exhibited in vitro antifungal activity against Alternaria sp., Aspergillus flavus, Aspergillus fumigatus, Curvularia lunata, Exophiala jeanselmei, Fonsecaea pedrosoi, Paecilomyces variotii, and Scedosporium apiospermum. The drug appeared to lack significant in vitro inhibitory activity against Fusarium oxysporum, Fusarium solani, Rhizopus arrhizus, Paecilomyces lilacinus, and Scedosporium prolificans. PMID:9257774

  16. [Importance of estrogens and estrogen-active compounds for udder health in cattle. A review].

    PubMed

    Zdunczyk, S; Zerbe, H; Hoedemaker, M

    2003-11-01

    High oestrogen concentrations in blood or high intake of oestrogen-active compounds with forage can be associated with an enhanced occurrence of udder diseases. Mean somatic cell count (MSCC) can increase and milk yield can decrease. Subclinically infected udder quarters can develop clinical mastitis and the rate of new infections can be high. This review describes concentrations of oestrogens in peripheral blood plasma in cattle and occurrence of oestrogen-active compounds in forage. Relationships between oestrogens or oestrogen-active compounds and udder health are presented. The possible mechanisms of enhanced susceptibility of the udder to infection under the influence of oestrogens are discussed. PMID:14679840

  17. Structural transitions during prothrombin activation: On the importance of fragment 2

    PubMed Central

    Adams, Ty E.; Huntington, James A.

    2016-01-01

    Prothrombin is activated to thrombin by the prothrombinase complex through sequential cleavage at two distinct sites. This occurs at sites of vascular injury in a highly regulated cascade of serine protease and cofactor activation, where activated platelets provide a suitable surface for protease/cofactor/substrate assembly. The precise structural and conformational changes undergone during the transition from prothrombin to thrombin have been studied for decades, and several structures of prothrombin fragments along the activation pathway have been solved. Here we present a new structure analyzed in context of other recent structures and biochemical studies. What emerges is an unexpected mechanism that involves a change in the mode of binding of the F2 domain (fragment 2) on the catalytic domain after cleavage at Arg320, and a subsequent reorientation of the linker between the F2 and catalytic domain to present the Arg271 site for cleavage. PMID:26365066

  18. The importance of behavior theory in control system modeling of physical activity sensor data.

    PubMed

    Riley, William T; Martin, Cesar A; Rivera, Daniel E

    2014-01-01

    Among health behaviors, physical activity has the most extensive record of research using passive sensors. Control systems and other system dynamic approaches have long been considered applicable for understanding human behavior, but only recently has the technology provided the precise and intensive longitudinal data required for these analytic approaches. Although sensors provide intensive data on the patterns and variations of physical activity over time, the influences of these variations are often unmeasured. Health behavior theories provide an explanatory framework of the putative mediators of physical activity changes. Incorporating the intensive longitudinal measurement of these theoretical constructs is critical to improving the fit of control system model of physical activity and for advancing behavioral theory. Theory-based control models also provide guidance on the nature of the controllers which serve as the basis for just-in-time adaptive interventions based on these control system models.

  19. Activity of Bifenthrin, Chlorfenapyr, Fipronil, and Thiamethoxam against Red Imported Fire Ants (Hymenoptera: Formicidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bifenthrin, chlorfenapyr, fipronil, and thiamethoxam were evaluated for activity against the Argentine ant, Linepithema humile (Mayr). Mobility impairment and lethal times were determined following topical treatments. Ants were immobilized most quickly by bifenthrin, followed by chlorfenapyr and thi...

  20. The importance of behavior theory in control system modeling of physical activity sensor data.

    PubMed

    Riley, William T; Martin, Cesar A; Rivera, Daniel E

    2014-01-01

    Among health behaviors, physical activity has the most extensive record of research using passive sensors. Control systems and other system dynamic approaches have long been considered applicable for understanding human behavior, but only recently has the technology provided the precise and intensive longitudinal data required for these analytic approaches. Although sensors provide intensive data on the patterns and variations of physical activity over time, the influences of these variations are often unmeasured. Health behavior theories provide an explanatory framework of the putative mediators of physical activity changes. Incorporating the intensive longitudinal measurement of these theoretical constructs is critical to improving the fit of control system model of physical activity and for advancing behavioral theory. Theory-based control models also provide guidance on the nature of the controllers which serve as the basis for just-in-time adaptive interventions based on these control system models. PMID:25571577

  1. Chemical and biological mechanisms of phytochemical activation of Nrf2 and importance in disease prevention

    PubMed Central

    Eggler, Aimee L.; Savinov, Sergey N.

    2016-01-01

    Plants are an incredibly rich source of compounds that activate the Nrf2 transcription factor, leading to upregulation of a battery of cytoprotective genes. This perspective surveys established and proposed molecular mechanisms of Nrf2 activation by phytochemicals with a special emphasis on a common chemical property of Nrf2 activators: the ability as “soft” electrophiles to modify cellular thiols, either directly or as oxidized biotransformants. In addition, the role of reactive oxygen/nitrogen species as secondary messengers in Nrf2 activation is discussed. While the uniquely reactive C151 of Keap1, an Nrf2 repressor protein, is highlighted as a key target of cytoprotective phytochemicals, also reviewed are other stress-responsive proteins, including kinases, which play non-redundant roles in the activation of Nrf2 by plant-derived agents. Finally, the perspective presents two key factors accounting for the enhanced therapeutic windows of effective phytochemical activators of the Keap1–Nrf2 axis: enhanced selectivity toward sensor cysteines and reversibility of addition to thiolate molecules. PMID:26855455

  2. Affinity Is an Important Determinant of the Anti-Trypanosome Activity of Nanobodies

    PubMed Central

    Caljon, Guy; Stijlemans, Benoît; Saerens, Dirk; Van Den Abbeele, Jan; Muyldermans, Serge; Magez, Stefan; De Baetselier, Patrick

    2012-01-01

    Background The discovery of Nanobodies (Nbs) with a direct toxic activity against African trypanosomes is a recent advancement towards a new strategy against these extracellular parasites. The anti-trypanosomal activity relies on perturbing the highly active recycling of the Variant-specific Surface Glycoprotein (VSG) that occurs in the parasite's flagellar pocket. Methodology/Principal Findings Here we expand the existing panel of Nbs with anti-Trypanosoma brucei potential and identify four categories based on their epitope specificity. We modified the binding properties of previously identified Nanobodies Nb_An05 and Nb_An33 by site-directed mutagenesis in the paratope and found this to strongly affect trypanotoxicity despite retention of antigen-targeting properties. Affinity measurements for all identified anti-trypanosomal Nbs reveal a strong correlation between trypanotoxicity and affinity (KD), suggesting that it is a crucial determinant for this activity. Half maximal effective (50%) affinity of 57 nM was calculated from the non-linear dose-response curves. In line with these observations, Nb humanizing mutations only preserved the trypanotoxic activity if the KD remained unaffected. Conclusions/Significance This study reveals that the binding properties of Nanobodies need to be compatible with achieving an occupancy of >95% saturation of the parasite surface VSG in order to exert an anti-trypanosomal activity. As such, Nb-based approaches directed against the VSG target would require binding to an accessible, conserved epitope with high affinity. PMID:23166849

  3. P38 activation is more important than ERK activation in lung injury induced by prolonged hyperbaric oxygen.

    PubMed

    Ma, Jun; Fang, Yi-Qun; Gu, Ai-Mei; Wang, Fang-Fang; Zhang, Shi; Li, Kai-Cheng

    2013-01-01

    Prolonged exposure to hyperbaric oxygen can cause pulmonary and nerve system toxicity. Although hyperbaric oxygen treatment has been used for a broad spectrum of ailments, the mechanisms of prolonged hyperbaric oxygen-induced lung injury are not fully understood. The purpose of the present work was to investigate the roles of ERK, p38, and caspase-3 in rat lung tissue exposed to hyperbaric oxygen at 2.3 atmospheres absolute (atm abs) for two, six and 10 hours. The results showed that the ERK and p38 were phosphorylated at two hours and reached a peak at six hours into exposure to hyperbaric oxygen. While the phosphorylation level of ERK decreased, p38 remained at a high level of activation at 10 hours. The activation of ERK and p38 was down-regulated when rats were exposed to normoxic hyperbaric nitrogen for 10 hours. However, caspase-3 was activated at six hours and 10 hours into exposure to hyperbaric oxygen. These results demonstrated different changes of activation of ERK and p38 during lung injury induced by prolonged exposure to hyperbaric oxygen. The time course changes of activated caspase-3 were similar to the process of p38 activation upon exposure to hyperbaric oxygen. In this way, activation of p38, not ERK, seems to be a mechanism associated with prolonged hyperbaric oxygen-induced lung injury.

  4. Active faults crossing trunk pipeline routes: some important steps to avoid the disaster

    NASA Astrophysics Data System (ADS)

    Besstrashnov, Vladimir; Strom, Alexander

    2010-05-01

    Trunk pipelines that pass through tectonically active areas connecting oil and gas reservoirs with terminals and refineries cross active faults that can produce large earthquakes. Besides strong motion affecting vast areas, these earthquakes are often associated with surface faulting that provides additional hazard to pipelines. To avoid significant economic losses and environmental pollution, pipelines should be designed to sustain both effects (shaking and direct rupturing) without pipe damage and spill. Special studies aimed to provide necessary input data for the designers should be performed in the course of engineering survey. However, our experience on conducting and review of such studies for several oil and gas trunk pipelines in Russia show urgent need of more strict definition of basic conceptions and approaches used for identification and localization of these potentially hazardous tectonic features. Identification of active faults (fault zones) considered as causative faults - sources of strong motion caused by seismic waves that affect dozens kilometers of pipeline route can be done by use of both direct and indirect evidence of Late Pleistocene - Holocene activity of faults and fault zones. Since strong motion parameters can be considered as constant within the near-field zone, which width in case of large earthquake is up to dozens kilometers, accuracy of active fault location is not so critical and ±1-2 km precision provided by use of indirect evidence is acceptable. In contrast, if one have to identify and characterize zones of potential surface rupturing that require special design of the endangered pipeline section, only direct evidence of such activity can provide reliable input data for crossing design with relevant accuracy of fault location, amount and direction of displacement. Only traces of surface faults displacing Late Pleistocene - Holocene sediments and/or geomorphic features are considered as direct evidence of fault activity. Just

  5. A Qualitative Study of Environmental Factors Important for Physical Activity in Rural Adults

    PubMed Central

    Cleland, Verity; Hughes, Clarissa; Thornton, Lukar; Venn, Alison; Squibb, Kathryn; Ball, Kylie

    2015-01-01

    Purpose Despite increasing evidence that the physical environment impacts on physical activity among urban-dwellers, little attention has been devoted to understanding this relationship in rural populations. Work in this area is further hindered by a lack of environmental measures specifically designed for rural settings. This qualitative study aimed to explore the salience of urban physical activity environment constructs among rural adults. Methods In 2011, 49 rural men and women from three distinct areas (coastal, animal-based farming, forestry/plant-based farming) of rural Tasmania, Australia, were purposively recruited to participate in semi-structured interviews. Interviews explored features of the built and social environment commonly examined in studies of urban adults, including functional characteristics (eg, lighting, footpaths, roads/verges), road and personal safety, availability and accessibility of places to be active, destinations, and aesthetics. Interviews were recorded, transcribed verbatim and analysed using a content-thematic approach using QSR NVivo software. Findings While some urban environmental constructs were salient to these rural adults, such as availability of and accessibility to places to be active, some constructs were operationalised differently, such as road safety (where large trucks and winding roads rather than traffic density was of concern), or were not considered relevant (eg, personal safety related to crime, availability of walkable destinations, aesthetics). Conclusions The measurement of the physical environment in rural populations may require reconsideration and/or modification to ensure salience and appropriate quantification of associations with physical activity in future studies. PMID:26554376

  6. High Sulfation and a High Molecular Weight Are Important for Anti-hepcidin Activity of Heparin.

    PubMed

    Asperti, Michela; Naggi, Annamaria; Esposito, Emiliano; Ruzzenenti, Paola; Di Somma, Margherita; Gryzik, Magdalena; Arosio, Paolo; Poli, Maura

    2015-01-01

    Heparins are efficient inhibitors of hepcidin expression even in vivo, where they induce an increase of systemic iron availability. Heparins seem to act by interfering with BMP6 signaling pathways that control the expression of liver hepcidin, causing the suppression of SMAD1/5/8 phosphorylation. The anti-hepcidin activity persists also when the heparin anticoagulant property is abolished or reduced by chemical reactions of oxidation/reduction (glycol-split, Gs-Heparins) or by high sulfation (SS-Heparins), but the structural characteristics needed to optimize this inhibitory activity have not been studied in detail. To this aim we analyzed three different heparins (Mucosal Heparin, the Glycol split RO-82, the partially desulfated glycol-split RO-68 and the oversulfated SSLMWH) and separated them in fractions of molecular weight in the range 4-16 kD. Since the distribution of the negative charges in heparins contributes to the activity, we produced 2-O- and 6-O-desulfated heparins. These derivatives were analyzed for the capacity to inhibit hepcidin expression in hepatic HepG2 cells and in mice. The two approaches produced consistent results and showed that the anti-hepcidin activity strongly decreases with molecular weight below 7 kD, with high N-acetylation and after 2-O and 6-O desulfation. The high sulfation and high molecular weight properties for efficient anti-hepcidin activity suggest that heparin is involved in multiple binding sites. PMID:26955355

  7. Pomegranate juice and prostate cancer: importance of the characterisation of the active principle.

    PubMed

    Chrubasik-Hausmann, Sigrun; Vlachojannis, Christian; Zimmermann, Benno

    2014-11-01

    Two exploratory clinical studies investigating proprietary pomegranate products showed a trend of effectiveness in increasing prostate-specific antigen doubling time in patients with prostate cancer. A recent clinical study did not support these results. We therefore analysed a lot of the marketed pomegranate blend for co-active pomegranate compounds. The high-performance liquid chromatography method was used to detect punicalagin, ellagic acid and anthocyanins. Total polyphenoles were determined by the Folin-Ciocalteu method using gallic acid as reference. The results show that the co-active compounds in the daily dose of the pomegranate blend were far below those previously tested and that the photometric assessment is not reliable for the standardisation of study medications. Not pomegranate but the low amount of co-active compounds in the proprietary pomegranate blend was responsible for its clinical ineffectiveness.

  8. The importance of physical activity in osteoporosis. From the molecular pathways to the clinical evidence.

    PubMed

    Castrogiovanni, Paola; Trovato, Francesca Maria; Szychlinska, Marta Anna; Nsir, Houda; Imbesi, Rosa; Musumeci, Giuseppe

    2016-11-01

    Osteoporosis is a very common bone disorder characterized by low bone mass and signs of deterioration, responsible for bone fragility typical in this pathology. The risk factors for the onset of osteoporosis are many and different from each other. Some of them cannot be modified, such as age, hereditary diseases and endocrine diseases. Others are modifiable, so that prevention is an advisable tool to reduce the incidence of osteoporosis. Among preventive tools, physical activity is certainly a valid instrument of prevention, in fact physical activity contributes to a healthy energy balance and increases muscle mass and bone mass. In the present narrative review, we wanted to pay attention to the possible influence of physical activity on the pathophysiological molecular pathways of osteoporosis and to the use of different exercise training in treatment of osteoporosis. From the literature analyzed, in relation to the effects of physical activity on bone metabolism, it is shown that exercise acts on molecular pathways of bone remodeling involving all cellular types of bone tissue. In relation to clinical trials adopted in patients with osteoporosis, it is evident that a multi-component training, including aerobic activity and other types of training (resistance and/or strength exercises), is the best kind of exercise in improving bone mass and bone metabolism in older adults and especially osteopoenic and osteoporotic women. With regard to whole-body-vibration training, it seems to be a valid alternative to current methods due to its greater adaptability to patients. In conclusion, physical activity, whatever the adopted training, always has beneficial effects on patients suffering from osteoporosis, and not only on bone homeostasis but on the whole skeletal muscle system.

  9. Physiological community ecology: variation in metabolic activity of ecologically important rocky intertidal invertebrates along environmental gradients.

    PubMed

    Dahlhoff, Elizabeth P; Stillman, Jonathon H; Menge, Bruce A

    2002-08-01

    Rocky intertidal invertebrates live in heterogeneous habitats characterized by steep gradients in wave activity, tidal flux, temperature, food quality and food availability. These environmental factors impact metabolic activity via changes in energy input and stress-induced alteration of energetic demands. For keystone species, small environmentally induced shifts in metabolic activity may lead to disproportionately large impacts on community structure via changes in growth or survival of these key species. Here we use biochemical indicators to assess how natural differences in wave exposure, temperature and food availability may affect metabolic activity of mussels, barnacles, whelks and sea stars living at rocky intertidal sites with different physical and oceanographic characteristics. We show that oxygen consumption rate is correlated with the activity of key metabolic enzymes (e.g., citrate synthase and malate dehydrogenase) for some intertidal species, and concentrations of these enzymes in certain tissues are lower for starved individuals than for those that are well fed. We also show that the ratio of RNA to DNA (an index of protein synthetic capacity) is highly variable in nature and correlates with short-term changes in food availability. We also observed striking patterns in enzyme activity and RNA/DNA in nature, which are related to differences in rocky intertidal community structure. Differences among species and habitats are most pronounced in summer and are linked to high nearshore productivity at sites favored by suspension feeders and to exposure to stressful low-tide air temperatures in areas of low wave splash. These studies illustrate the great promise of using biochemical indicators to test ecological models, which predict changes in community structure along environmental gradients. Our results also suggest that biochemical indices must be carefully validated with laboratory studies, so that the indicator selected is likely to respond to the

  10. Physiological community ecology: variation in metabolic activity of ecologically important rocky intertidal invertebrates along environmental gradients.

    PubMed

    Dahlhoff, Elizabeth P; Stillman, Jonathon H; Menge, Bruce A

    2002-08-01

    Rocky intertidal invertebrates live in heterogeneous habitats characterized by steep gradients in wave activity, tidal flux, temperature, food quality and food availability. These environmental factors impact metabolic activity via changes in energy input and stress-induced alteration of energetic demands. For keystone species, small environmentally induced shifts in metabolic activity may lead to disproportionately large impacts on community structure via changes in growth or survival of these key species. Here we use biochemical indicators to assess how natural differences in wave exposure, temperature and food availability may affect metabolic activity of mussels, barnacles, whelks and sea stars living at rocky intertidal sites with different physical and oceanographic characteristics. We show that oxygen consumption rate is correlated with the activity of key metabolic enzymes (e.g., citrate synthase and malate dehydrogenase) for some intertidal species, and concentrations of these enzymes in certain tissues are lower for starved individuals than for those that are well fed. We also show that the ratio of RNA to DNA (an index of protein synthetic capacity) is highly variable in nature and correlates with short-term changes in food availability. We also observed striking patterns in enzyme activity and RNA/DNA in nature, which are related to differences in rocky intertidal community structure. Differences among species and habitats are most pronounced in summer and are linked to high nearshore productivity at sites favored by suspension feeders and to exposure to stressful low-tide air temperatures in areas of low wave splash. These studies illustrate the great promise of using biochemical indicators to test ecological models, which predict changes in community structure along environmental gradients. Our results also suggest that biochemical indices must be carefully validated with laboratory studies, so that the indicator selected is likely to respond to the

  11. The Relative Importance of Academic Activities: Autonomous Values from the Canadian Professoriate

    ERIC Educational Resources Information Center

    Osakwe, Carlton; Keavey, Kalen; Uzoka, Faith-Michael; Fedoruk, Alan; Osuji, Joseph

    2015-01-01

    In this paper, we report findings on the contemporary idea of a university as defined by the relative importance of teaching, research, service, and collegiality in teaching-intensive universities. We also investigate Boyer's model of scholarship relative to the research--teaching--service nexus. Our findings show that while research is embraced…

  12. 75 FR 67981 - Agency Information Collection Activities; Proposed Collection; Comment Request; Importer's Entry...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-11-04

    ... this collection of information expands the universe of respondents being regulated under the FD&C Act... origin FDA regulated foods, drugs, cosmetics, medical devices, radiological health, and tobacco products..., landport, and airport) where foreign-origin FDA-regulated products are offered for import, FDA is...

  13. Importance of Health-Related Fitness Knowledge to Increasing Physical Activity and Physical Fitness

    ERIC Educational Resources Information Center

    Ferkel, Rick C.; Judge, Lawrence W.; Stodden, David F.; Griffin, Kent

    2014-01-01

    Physical inactivity is expanding across all ages in the United States. Research has documented a deficiency in health-related fitness knowledge (HRFK) among elementary- through college-aged students. The need for a credible and reliable resource that provides research-based information regarding the importance of HRFK is significant. The purpose…

  14. 78 FR 19312 - Agency Information Collection Activities; Proposed Collection; Comments Requested: Import/Export...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-03-29

    ...: Import/Export Declaration for List I and List II Chemicals, DEA Forms 486 and 486A ACTION: 60-Day Notice. The Department of Justice (DOJ), Drug Enforcement Administration (DEA), will be submitting the... the Department of Justice sponsoring the collection: Form Number: DEA Forms 486 and 486A....

  15. 77 FR 62532 - Agency Information Collection Activities; Proposed Collection; Comments Requested: Import/Export...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-15

    ...: Import/Export Declaration for List I and List II Chemicals; DEA Forms 486 and 486A ACTION: 30-Day Notice. The Department of Justice (DOJ), Drug Enforcement Administration (DEA) will be submitting the... Department sponsoring the collection: Form number: DEA Forms 486 and 486A. Component: Office of...

  16. 77 FR 47666 - Agency Information Collection Activities: Proposed Collection; Comments Requested: Import/Export...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-08-09

    ...: Import/Export Declaration for List I and List II Chemicals; DEA Forms 486 and 486A ACTION: 60-Day Notice... Administration (DEA), will be submitting the following information collection request to the Office of Management... component of the Department of Justice sponsoring the collection: Form Number: DEA Forms 486 and...

  17. What is the importance of abnormal "background" activity in seizure generation?

    PubMed

    Staba, Richard J; Worrell, Gregory A

    2014-01-01

    Investigations of interictal epileptiform spikes and seizures have played a central role in the study of epilepsy. The background EEG activity, however, has received less attention. In this chapter we discuss the characteristic features of the background activity of the brain when individuals are at rest and awake (resting wake) and during sleep. The characteristic rhythms of the background EEG are presented, and the presence of 1/f (β) behavior of the EEG power spectral density is discussed and its possible origin and functional significance. The interictal EEG findings of focal epilepsy and the impact of interictal epileptiform spikes on cognition are also discussed. PMID:25012365

  18. Biological and chemical study of fused tri- and tetracyclic indazoles and analogues with important antiparasitic activity

    NASA Astrophysics Data System (ADS)

    Díaz-Urrutia, Christian A.; Olea-Azar, Claudio A.; Zapata, Gerald A.; Lapier, Michel; Mura, Francisco; Aguilera-Venegas, Benjamín; Arán, Vicente J.; López-Múñoz, Rodrigo A.; Maya, Juan D.

    A series of fused tri- and tetracyclic indazoles and analogues compounds (NID) with potential antiparasitic effects were studied using voltamperometric and spectroscopic techniques. Nitroanion radicals generated by cyclic voltammetry were characterized by electron spin resonance spectroscopy (ESR) and their spectral lines were explained and analyzed using simulated spectra. In addition, we examined the interaction between radical species generated from nitroindazole derivatives and glutathione (GSH). Biological assays such as activity against Trypanosoma cruzi and cytotoxicity against macrophages were carried out. Finally, spin trapping and molecular modeling studies were also done in order to elucidate the potentials action mechanisms involved in the trypanocidal activity.

  19. Biological and chemical study of fused tri- and tetracyclic indazoles and analogues with important antiparasitic activity.

    PubMed

    Díaz-Urrutia, Christian A; Olea-Azar, Claudio A; Zapata, Gerald A; Lapier, Michel; Mura, Francisco; Aguilera-Venegas, Benjamín; Arán, Vicente J; López-Múñoz, Rodrigo A; Maya, Juan D

    2012-09-01

    A series of fused tri- and tetracyclic indazoles and analogues compounds (NID) with potential antiparasitic effects were studied using voltamperometric and spectroscopic techniques. Nitroanion radicals generated by cyclic voltammetry were characterized by electron spin resonance spectroscopy (ESR) and their spectral lines were explained and analyzed using simulated spectra. In addition, we examined the interaction between radical species generated from nitroindazole derivatives and glutathione (GSH). Biological assays such as activity against Trypanosoma cruzi and cytotoxicity against macrophages were carried out. Finally, spin trapping and molecular modeling studies were also done in order to elucidate the potentials action mechanisms involved in the trypanocidal activity.

  20. Resisting Rote: The Importance of Active Learning for All Course Learning Objectives

    ERIC Educational Resources Information Center

    Fata-Hartley, Cori

    2011-01-01

    Many college science educators have moved away from the traditional lecture format and toward learner-centered classroom environments. Yet many of us struggle to cover large content loads, reverting at times to rote memorization. This paper suggests rote memorization simply does not work and students must be actively engaged to learn. (Contains 1…

  1. The importance of physical activity and sleep for affect on stressful days: Two intensive longitudinal studies.

    PubMed

    Flueckiger, Lavinia; Lieb, Roselind; Meyer, Andrea H; Witthauer, Cornelia; Mata, Jutta

    2016-06-01

    We investigated the potential stress-buffering effect of 3 health behaviors-physical activity, sleep quality, and snacking-on affect in the context of everyday life in young adults. In 2 intensive longitudinal studies with up to 65 assessment days over an entire academic year, students (Study 1, N = 292; Study 2, N = 304) reported stress intensity, sleep quality, physical activity, snacking, and positive and negative affect. Data were analyzed using multilevel regression analyses. Stress and positive affect were negatively associated; stress and negative affect were positively associated. The more physically active than usual a person was on a given day, the weaker the association between stress and positive affect (Study 1) and negative affect (Studies 1 and 2). The better than usual a person's sleep quality had been during the previous night, the weaker the association between stress and positive affect (Studies 1 and 2) and negative affect (Study 2). The association between daily stress and positive or negative affect did not differ as a function of daily snacking (Studies 1 and 2). On stressful days, increasing physical activity or ensuring high sleep quality may buffer adverse effects of stress on affect in young adults. These findings suggest potential targets for health-promotion and stress-prevention programs, which could help reduce the negative impact of stress in young adults. (PsycINFO Database Record

  2. Apoptosome activation, an important molecular instigator in 6-mercaptopurine induced Leydig cell death

    PubMed Central

    Morgan, Jessica A.; Lynch, John; Panetta, John C.; Wang, Yao; Frase, Sharon; Bao, Ju; Zheng, Jie; Opferman, Joseph T.; Janke, Laura; Green, Daniel M.; Chemaitilly, Wassim; Schuetz, John D.

    2015-01-01

    Leydig cells are crucial to the production of testosterone in males. It is unknown if the cancer chemotherapeutic drug, 6-mercaptopurine (6 MP), produces Leydig cell failure among adult survivors of childhood acute lymphoblastic leukemia. Moreover, it is not known whether Leydig cell failure is due to either a loss of cells or an impairment in their function. Herein, we show, in a subset of childhood cancer survivors, that Leydig cell failure is related to the dose of 6 MP. This was extended, in a murine model, to demonstrate that 6 MP exposure induced caspase 3 activation, and the loss of Leydig cells was independent of Bak and Bax activation. The death of these non-proliferating cells was triggered by 6 MP metabolism, requiring formation of both cytosolic reactive oxygen species and thiopurine nucleotide triphosphates. The thiopurine nucleotide triphosphates (with physiological amounts of dATP) uniquely activated the apoptosome. An ABC transporter (Abcc4/Mrp4) reduced the amount of thiopurines, thereby providing protection for Leydig cells. The studies reported here demonstrate that the apoptosome is uniquely activated by thiopurine nucleotides and suggest that 6 MP induced Leydig cell death is likely a cause of Leydig cell failure in some survivors of childhood cancer. PMID:26576726

  3. Ambulatory Activity of Children with Cerebral Palsy: Which Characteristics Are Important?

    ERIC Educational Resources Information Center

    van Wely, Leontien; Becher, Jules G.; Balemans, Astrid C. J.; Dallmeijer, Annet J.

    2012-01-01

    Aim: To assess ambulatory activity of children with cerebral palsy (CP), aged 7 to 13 years, and identify associated characteristics. Method: Sixty-two children with spastic CP (39 males, 23 females; mean age 10y 1mo, SD 1y 8mo; age range 7-13y), classified as Gross Motor Function Classification System (GMFCS) levels I to III, participated.…

  4. Antiausterity activity of arctigenin enantiomers: importance of (2R,3R)-absolute configuration.

    PubMed

    Awale, Suresh; Kato, Mamoru; Dibwe, Dya Fita; Li, Feng; Miyoshi, Chika; Esumi, Hiroyasu; Kadota, Shigetoshi; Tezuka, Yasuhiro

    2014-01-01

    From a MeOH extract of powdered roots of Wikstroemia indica, six dibenzyl-gamma-butyrolactone-type lignans with (2S,3S)-absolute configuration [(+)-arctigenin (1), (+)-matairesinol (2), (+)-trachelogenin (3), (+)-nortrachelogenin (4), (+)-hinokinin (5), and (+)-kusunokinin (6)] were isolated, whereas three dibenzyl-gamma-butyrolactone-type lignans with (2R,3R)-absolute configuration [(-)-arctigenin (1*), (-)-matairesinol (2*), (-)-trachelogenin (3*)] were isolated from Trachelospermum asiaticum. The in vitro preferential cytotoxic activity of the nine compounds was evaluated against human pancreatic PANC-1 cancer cells in nutrient-deprived medium (NDM), but none of the six lignans (1-6) with (2S,3S)-absolute configuration showed preferential cytotoxicity. On the other hand, three lignans (1*-3*) with (2R,3R)-absolute configuration exhibited preferential cytotoxicity in a concentration-dependent manner with PC50 values of 0.54, 6.82, and 5.85 microM, respectively. Furthermore, the effect of (-)- and (+)-arctigenin was evaluated against the activation of Akt, which is a key process in the tolerance to nutrition starvation. Interestingly, only (-)-arctigenin (1*) strongly suppressed the activation of Akt. These results indicate that the (2R,3R)-absolute configuration of (-)-enantiomers should be required for the preferential cytotoxicity through the inhibition of Akt activation.

  5. Adolescents Online: The Importance of Internet Activity Choices to Salient Relationships

    ERIC Educational Resources Information Center

    Blais, Julie J.; Craig, Wendy M.; Pepler, Debra; Connolly, Jennifer

    2008-01-01

    The purpose of this study was to determine whether using the Internet for different activities affects the quality of close adolescent relationships (i.e., best friendships and romantic relationships). In a one-year longitudinal study of 884 adolescents (Mean age = 15, 46% male), we examined whether visiting chat rooms, using ICQ, using the…

  6. 75 FR 45527 - Taking and Importing Marine Mammals; Military Training Activities and Research, Development...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-03

    ... under ] the MMPA: Blue whale; fin whale; humpback whale; sei whale; sperm whale; North Pacific right... activity, which was published in the Federal Register on October 20, 2009 (74 FR 53796). This information... individuals of 2 species by Level A Harassment annually, and 10 individual beaked whales by mortality over...

  7. Important Physiological Parameters and Physical Activity Data for Evaluating Exposure Modeling Performance: a Synthesis

    EPA Science Inventory

    The purpose of this report is to develop a database of physiological parameters needed for understanding and evaluating performance of the APEX and SHEDS exposure/intake dose rate model used by the Environmental Protection Agency (EPA) as part of its regulatory activities. The A...

  8. Importance of Quality Recreation Activities for Older Adults Residing in Nursing Homes: Considerations for Gerontologists.

    ERIC Educational Resources Information Center

    Haberkost, Michael; And Others

    1996-01-01

    Based on a needs assessment survey (66 responses from 101 nursing home activity coordinators), a recreation manual and training program was developed and tested with 25 coordinators/recreation staff. The 14 who completed evaluations increased their understanding of such topics as depression; goals of nursing home recreation programs; motivation of…

  9. Antiausterity activity of arctigenin enantiomers: importance of (2R,3R)-absolute configuration.

    PubMed

    Awale, Suresh; Kato, Mamoru; Dibwe, Dya Fita; Li, Feng; Miyoshi, Chika; Esumi, Hiroyasu; Kadota, Shigetoshi; Tezuka, Yasuhiro

    2014-01-01

    From a MeOH extract of powdered roots of Wikstroemia indica, six dibenzyl-gamma-butyrolactone-type lignans with (2S,3S)-absolute configuration [(+)-arctigenin (1), (+)-matairesinol (2), (+)-trachelogenin (3), (+)-nortrachelogenin (4), (+)-hinokinin (5), and (+)-kusunokinin (6)] were isolated, whereas three dibenzyl-gamma-butyrolactone-type lignans with (2R,3R)-absolute configuration [(-)-arctigenin (1*), (-)-matairesinol (2*), (-)-trachelogenin (3*)] were isolated from Trachelospermum asiaticum. The in vitro preferential cytotoxic activity of the nine compounds was evaluated against human pancreatic PANC-1 cancer cells in nutrient-deprived medium (NDM), but none of the six lignans (1-6) with (2S,3S)-absolute configuration showed preferential cytotoxicity. On the other hand, three lignans (1*-3*) with (2R,3R)-absolute configuration exhibited preferential cytotoxicity in a concentration-dependent manner with PC50 values of 0.54, 6.82, and 5.85 microM, respectively. Furthermore, the effect of (-)- and (+)-arctigenin was evaluated against the activation of Akt, which is a key process in the tolerance to nutrition starvation. Interestingly, only (-)-arctigenin (1*) strongly suppressed the activation of Akt. These results indicate that the (2R,3R)-absolute configuration of (-)-enantiomers should be required for the preferential cytotoxicity through the inhibition of Akt activation. PMID:24660468

  10. Physical Activity Based Professional Development for Teachers: The Importance of Whole School Involvement

    ERIC Educational Resources Information Center

    Till, Jude; Ferkins, Lesley; Handcock, Phil

    2011-01-01

    Objective: This study sought to investigate teachers' perceptions of a physical activity-related professional development intervention. Design: Interview-based qualitative approach founded on the interpretive paradigm. Setting: Purposive selection of one high-rated independent, and one low-rated public primary school from Auckland, New Zealand.…

  11. Student Perceptions of Information Literacy Instruction: The Importance of Active Learning

    ERIC Educational Resources Information Center

    Detlor, Brian; Booker, Lorne; Serenko, Alexander; Julien, Heidi

    2012-01-01

    This study investigates the merits of employing active learning strategies in the delivery of information literacy instruction (ILI). Traditional approaches to the teaching of information literacy skills--where students are passive recipients of the information they receive--are challenged. Rather, methods that encourage students to actively…

  12. 76 FR 50457 - Taking and Importing Marine Mammals; Military Training Activities and Research Conducted Within...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-15

    ... became effective on August 3, 2010 (75 FR 45527, August 3, 2010), and remain in effect through August 3... under regulations issued on August 3, 2010 (75 FR 45527). The Navy has complied with the measures...; Military Training Activities and Research Conducted Within the Mariana Islands Range Complex...

  13. Bling My Research! A Mock Grant Panel Activity Illustrating the Importance of Basic Research

    ERIC Educational Resources Information Center

    Leander, Celeste A.; Whitton, Jeannette

    2010-01-01

    First-year university students have misconceptions about the source and dynamics of publicly funded research money. We designed an activity in which students take part in a mock grant panel. The results indicated a strong tendency toward student funding of applied medical research at the expense of basic research. Exposure to a few examples of…

  14. C-Terminus of the B-Chain of Relaxin-3 Is Important for Receptor Activity

    PubMed Central

    Shabanpoor, Fazel; Bathgate, Ross A. D.

    2013-01-01

    Human relaxin-3 is a neuropeptide that is structurally similar to human insulin with two chains (A and B) connected by three disulfide bonds. It is expressed primarily in the brain and has modulatory roles in stress and anxiety, feeding and metabolism, and arousal and behavioural activation. Structure-activity relationship studies have shown that relaxin-3 interacts with its cognate receptor RXFP3 primarily through its B-chain and that its A-chain does not have any functional role. In this study, we have investigated the effect of modification of the B-chain C-terminus on the binding and activity of the peptide. We have chemically synthesised and characterized H3 relaxin as C-termini acid (both A and B chains having free C-termini; native form) and amide forms (both chains’ C-termini were amidated). We have confirmed that the acid form of the peptide is more potent than its amide form at both RXFP3 and RXFP4 receptors. We further investigated the effects of amidation at the C-terminus of individual chains. We report here for the first time that amidation at the C-terminus of the B-chain of H3 relaxin leads to significant drop in the binding and activity of the peptide at RXFP3/RXFP4 receptors. However, modification of the A-chain C-terminus does not have any effect on the activity. We have confirmed using circular dichroism spectroscopy that there is no secondary structural change between the acid and amide form of the peptide, and it is likely that it is the local C-terminal carboxyl group orientation that is crucial for interacting with the receptors. PMID:24349312

  15. Mathematical modeling of the intracellular protein dynamics: the importance of active transport along microtubules.

    PubMed

    Szymańska, Zuzanna; Parisot, Martin; Lachowicz, Mirosław

    2014-12-21

    In this paper we propose a mathematical model of protein and mRNA transport inside a cell. The spatio-temporal model takes into account the active transport along microtubules in the cytoplasm as well as diffusion and is able to reproduce the oscillatory changes in protein concentration observed in many experimental data. In the model the protein and the mRNA interact with each other that allows us to classify the model as a simple gene regulatory network. The proposed model is generic and may be adapted to specific signaling pathways. On the basis of numerical simulations, we formulate a new hypothesis that the oscillatory dynamics is allowed by the mRNA active transport along microtubules from the nucleus to distant locations.

  16. Soft Skills: An Important Asset Acquired from Organizing Regional Student Group Activities

    PubMed Central

    de Ridder, Jeroen; Meysman, Pieter; Oluwagbemi, Olugbenga; Abeel, Thomas

    2014-01-01

    Contributing to a student organization, such as the International Society for Computational Biology Student Council (ISCB-SC) and its Regional Student Group (RSG) program, takes time and energy. Both are scarce commodities, especially when you are trying to find your place in the world of computational biology as a graduate student. It comes as no surprise that organizing ISCB-SC-related activities sometimes interferes with day-to-day research and shakes up your priority list. However, we unanimously agree that the rewards, both in the short as well as the long term, make the time spent on these extracurricular activities more than worth it. In this article, we will explain what makes this so worthwhile: soft skills. PMID:24992198

  17. Soft skills: an important asset acquired from organizing regional student group activities.

    PubMed

    de Ridder, Jeroen; Meysman, Pieter; Oluwagbemi, Olugbenga; Abeel, Thomas

    2014-07-01

    Contributing to a student organization, such as the International Society for Computational Biology Student Council (ISCB-SC) and its Regional Student Group (RSG) program, takes time and energy. Both are scarce commodities, especially when you are trying to find your place in the world of computational biology as a graduate student. It comes as no surprise that organizing ISCB-SC-related activities sometimes interferes with day-to-day research and shakes up your priority list. However, we unanimously agree that the rewards, both in the short as well as the long term, make the time spent on these extracurricular activities more than worth it. In this article, we will explain what makes this so worthwhile: soft skills. PMID:24992198

  18. Soft skills: an important asset acquired from organizing regional student group activities.

    PubMed

    de Ridder, Jeroen; Meysman, Pieter; Oluwagbemi, Olugbenga; Abeel, Thomas

    2014-07-01

    Contributing to a student organization, such as the International Society for Computational Biology Student Council (ISCB-SC) and its Regional Student Group (RSG) program, takes time and energy. Both are scarce commodities, especially when you are trying to find your place in the world of computational biology as a graduate student. It comes as no surprise that organizing ISCB-SC-related activities sometimes interferes with day-to-day research and shakes up your priority list. However, we unanimously agree that the rewards, both in the short as well as the long term, make the time spent on these extracurricular activities more than worth it. In this article, we will explain what makes this so worthwhile: soft skills.

  19. Relative importance of complement-mediated bactericidal and opsonic activity for protection against meningococcal disease.

    PubMed

    Granoff, Dan M

    2009-06-24

    Killing of Neisseria meningitidis can result from complement-mediated serum bactericidal activity (SBA) or opsonophagocytosis (OPA), or a combination of the two mechanisms. While SBA titers > or =1:4 confer protection, recent evidence suggests that this threshold titer may not be required. For example, the incidence of meningococcal disease declines between ages 1 and 4 years without evidence of acquisition of SBA titers > or =1:4. Meningococcal polysaccharide vaccination also elicited OPA and lowered the risk of disease in patients with late complement component deficiencies whose sera did not support SBA. Sera from healthy adults immunized with an outer membrane vesicle vaccine showed OPA killing of N. meningitidis with C6-depleted complement, and whole blood from complement-sufficient non-immunized adults with SBA titers <1:4 also frequently had killing activity. Collectively the data indicate that SBA titers <1:4 and/or vaccine-induced OPA can confer protection against meningococcal disease.

  20. Genetic dissection of the phospholipid hydroperoxidase activity of yeast gpx3 reveals its functional importance.

    PubMed

    Avery, Angela M; Willetts, Sylvia A; Avery, Simon V

    2004-11-01

    Saccharomyces cerevisiae expresses multiple phospholipid hydroperoxide glutathione peroxidase (PHGPx)-like proteins in the absence of a classical glutathione peroxidase (cGPx), providing a unique system for dissecting the roles of these enzymes in vivo. The Gpx3 (Orp1/PHGpx3) protein transduces the hydroperoxide signal to the transcription factor Yap1, a function that could account for most GPX-dependent phenotypes. To test this hypothesis and ascertain what functions of Gpx3 can be shared by cGPx-like enzymes, we constructed a novel cGPx-like yeast enzyme, cGpx3. We confirmed that the "gap" sequences conserved among cGPxs but absent from aligned PHGPx sequences are the principal cause of the structural and functional differences of these enzymes. Peroxidase activity against a cGPx substrate was high in the cGpx3 construct, which was multimeric and had a peroxidase catalytic mechanism distinct from Gpx3; but cGpx3 was defective for phospholipid hydroperoxidase and signaling activities. cGpx3 did not complement the sensitivity to lipid peroxidation of a gpxDelta mutant, and the resistance to lipid peroxidation conferred by Gpx3 was independent of Yap1, establishing a functional role for Gpx3 phospholipid hydroperoxidase activity. Using the comparison between cGpx3 and Gpx3 in conjunction with other constructs to probe lipid peroxidation as a toxicity mechanism, we also ascertained that lipid peroxidation-dependent processes are a principal cause of cellular cadmium toxicity. The results demonstrate that phospholipid hydroperoxidase and Yap1-mediated signaling activities of Gpx3 have independent functional roles, although both functions depend on the absence of cGPx-like subunit interaction sites, and the results resolve more clearly the potential drivers of the differential selective evolution of GPx-like enzymes. PMID:15337745

  1. Importance of the disulfide bridges in the antibacterial activity of human hepcidin.

    PubMed

    Hocquellet, Agnès; le Senechal, Caroline; Garbay, Bertrand

    2012-08-01

    Hepcidin was first identified as an antimicrobial peptide present in human serum and urine. It was later demonstrated that hepcidin is the long sought hormone that regulates iron homeostasis in mammals. The native peptide of 25 amino acids (Hepc25) contains four disulfide bridges that maintain a β-hairpin motif. The aim of the present study was to assess whether the intramolecular disulfide bridges are necessary for Hepc25 antimicrobial activity. We show that a synthetic peptide corresponding to human Hepc25, and which contains the four disulfide bridges, has an antibacterial activity against several strains of Gram-positive and Gram-negative bacteria. On the contrary, a synthetic peptide where all cysteines were replaced by alanines (Hepc25-Ala) had no detectable activity against the same strains of bacteria. In a further step, the mode of action of Hepc25 on Escherichia coli was studied. SYTOX Green uptake was used to assess bacterial membrane integrity. No permeabilization of the membrane was observed with Hepc25, indicating that this peptide does not kill bacteria by destroying their membranes. Gel retardation assay showed that the Hepc25 binds to DNA with high efficiency, and that this binding ability is dependent on the presence of the intramolecular disulfide bridges. Reduction of Hepc25 or replacement of the eight cysteines by alanine residues led to peptides that were no longer able to bind DNA in the in vitro assay. Altogether, these results demonstrate that Hepc25 should adopt a three-dimensional structure stabilized by the intramolecular disulfide bridges in order to have antibacterial activity.

  2. Importance of Membrane Structural Integrity for RPE65 Retinoid Isomerization Activity

    SciTech Connect

    Golczak, Marcin; Kiser, Philip D.; Lodowski, David T.; Maeda, Akiko; Palczewski, Krzysztof

    2010-04-05

    Regeneration of visual chromophore in the vertebrate visual cycle involves the retinal pigment epithelium-specific protein RPE65, the key enzyme catalyzing the cleavage and isomerization of all-trans-retinyl fatty acid esters to 11-cis-retinol. Although RPE65 has no predicted membrane spanning domains, this protein predominantly associates with microsomal fractions isolated from bovine retinal pigment epithelium (RPE). We have re-examined the nature of RPE65 interactions with native microsomal membranes by using extraction and phase separation experiments. We observe that hydrophobic interactions are the dominant forces that promote RPE65 association with these membranes. These results are consistent with the crystallographic model of RPE65, which features a large lipophilic surface that surrounds the entrance to the catalytic site of this enzyme and likely interacts with the hydrophobic core of the endoplasmic reticulum membrane. Moreover, we report a critical role for phospholipid membranes in preserving the retinoid isomerization activity and physical properties of RPE65. Isomerase activity measured in bovine RPE was highly sensitive to phospholipase A{sup 2} treatment, but the observed decline in 11-cis-retinol production did not directly reflect inhibition by products of lipid hydrolysis. Instead, a direct correlation between the kinetics of phospholipid hydrolysis and retinoid isomerization suggests that the lipid membrane structure is critical for RPE65 enzymatic activity. We also provide evidence that RPE65 operates in a multiprotein complex with retinol dehydrogenase 5 and retinal G protein-coupled receptor in RPE microsomes. Modifications in the phospholipid environment affecting interactions with these protein components may be responsible for the alterations in retinoid metabolism observed in phospholipid-depleted RPE microsomes. Thus, our results indicate that the enzymatic activity of native RPE65 strongly depends on its membrane binding and

  3. Estimation of effective doses to adult and pediatric patients from multislice computed tomography: A method based on energy imparted

    SciTech Connect

    Theocharopoulos, Nicholas; Damilakis, John; Perisinakis, Kostas; Tzedakis, Antonis; Karantanas, Apostolos; Gourtsoyiannis, Nicholas

    2006-10-15

    The purpose of this study is to provide a method and required data for the estimation of effective dose (E) values to adult and pediatric patients from computed tomography (CT) scans of the head, chest abdomen, and pelvis, performed on multi-slice scanners. Mean section radiation dose (d{sub m}) to cylindrical water phantoms of varying radius normalized over CT dose index free-in-air (CTDI{sub F}) were calculated for the head and body scanning modes of a multislice scanner with use of Monte Carlo techniques. Patients were modeled as equivalent water phantoms and the energy imparted ({epsilon}) to simulated pediatric and adult patients was calculated on the basis of measured CTDI{sub F} values. Body region specific energy imparted to effective dose conversion coefficients (E/{epsilon}) for adult male and female patients were generated from previous data. Effective doses to patients aged newborn to adult were derived for all available helical and axial beam collimations, taking into account age specific patient mass and scanning length. Depending on high voltage, body region, and patient sex, E/{epsilon} values ranged from 0.008 mSv/mJ for head scans to 0.024 mSv/mJ for chest scans. When scanned with the same technique factors as the adults, pediatric patients absorb as little as 5% of the energy imparted to adults, but corresponding effective dose values are up to a factor of 1.6 higher. On average, pediatric patients absorb 44% less energy per examination but have a 24% higher effective dose, compared with adults. In clinical practice, effective dose values to pediatric patients are 2.5 to 10 times lower than in adults due to the adaptation of tube current. A method is provided for the calculation of effective dose to adult and pediatric patients on the basis of individual patient characteristics such as sex, mass, dimensions, and density of imaged anatomy, and the technical features of modern multislice scanners. It allows the optimum selection of scanning

  4. Local origins impart conserved bone type-related differences in human osteoblast behaviour.

    PubMed

    Shah, M; Gburcik, V; Reilly, P; Sankey, R A; Emery, R J; Clarkin, C E; Pitsillides, A A

    2015-03-04

    Osteogenic behaviour of osteoblasts from trabecular, cortical and subchondral bone were examined to determine any bone type-selective differences in samples from both osteoarthritic (OA) and osteoporotic (OP) patients. Cell growth, differentiation; alkaline phosphatase (TNAP) mRNA and activity, Runt-related transcription factor-2 (RUNX2), SP7-transcription factor (SP7), bone sialoprotein-II (BSP-II), osteocalcin/bone gamma-carboxyglutamate (BGLAP), osteoprotegerin (OPG, TNFRSF11B), receptor activator of nuclear factor-κβ ligand (RANKL, TNFSF11) mRNA levels and proangiogenic vascular endothelial growth factor-A (VEGF-A) mRNA and protein release were assessed in osteoblasts from paired humeral head samples from age-matched, human OA/OP (n = 5/4) patients. Initial outgrowth and increase in cell number were significantly faster (p < 0.01) in subchondral and cortical than trabecular osteoblasts, in OA and OP, and this bone type-related differences were conserved despite consistently faster growth in OA. RUNX2/SP7 levels and TNAP mRNA and protein activity were, however, greater in trabecular than subchondral and cortical osteoblasts in OA and OP. BSP-II levels were significantly greater in trabecular and lowest in cortical osteoblasts in both OA and OP. In contrast, BGLAP levels showed divergent bone type-selective behaviour; highest in osteoblasts from subchondral origins in OA and trabecular origins in OP. We found virtually identical bone type-related differences, however, in TNFRSF11B:TNFSF11 in OA and OP, consistent with greater potential for paracrine effects on osteoclasts in trabecular osteoblasts. Subchondral osteoblasts (OA) exhibited highest VEGF-A mRNA levels and release. Our data indicate that human osteoblasts in trabecular, subchondral and cortical bone have inherent, programmed diversity, with specific bone type-related differences in growth, differentiation and pro-angiogenic potential in vitro.

  5. Yeast Cytosine Deaminase Mutants with Increased Thermostability Impart Sensitivity to 5-Fluorocytosine

    PubMed Central

    Stolworthy, Tiffany S.; Korkegian, Aaron M.; Willmon, Candice L.; Ardiani, Andressa; Cundiff, Jennifer; Stoddard, Barry L.; Black, Margaret E.

    2008-01-01

    SUMMARY Prodrug gene therapy (PGT) is a treatment strategy in which tumor cells are transfected with a 'suicide' gene that encodes a metabolic enzyme capable of converting a nontoxic prodrug into a potent cytotoxin. One of the most promising PGT enzymes is cytosine deaminase (CD), a microbial salvage enzyme that converts cytosine to uracil. CD also converts 5-fluorocytosine (5FC) to 5-fluorouracil (5FU), an inhibitor of DNA synthesis and RNA function. Over 150 studies of cytosine deaminase-mediated PGT applications have been reported since 2000, all using wild-type enzymes. However, various forms of cytosine deaminase are limited by inefficient turnover of 5FC and/or limited thermostability. In a previous study we stabilized and extended the half-life of yeast cytosine deaminase (yCD) by repacking of its hydrophobic core at several positions distant from the active site. Here we report that random mutagenesis of residues selected based on alignment with similar enzymes, followed by selection for enhanced sensitization to 5FC, also produces an enzyme variant (yCD-D92E) with elevated Tm values and increased activity half-life. The new mutation is located at the enzyme's dimer interface, indicating that independent mutational pathways can lead to an increase in the temperature that induces protein unfolding and aggregation in thermal denaturation experiments measured by circular dichroism spectroscopy, and an increase in the half-life of enzyme activity at physiological temperature, as well as more subtle effect on enzyme kinetics. Each independently derived set of mutations significantly improves the enzyme's performance in PGT assays both in cell culture and in animal models. PMID:18291415

  6. The importance of diet and physical activity in the treatment of conditions managed in general practice.

    PubMed Central

    Little, P; Margetts, B

    1996-01-01

    Evidence from meta-analyses, physiological data and individual studies suggests that diet and exercise are important in the aetiology and treatment of many of the conditions that are managed predominantly in primary care (hypercholesterolaemia, hypertension, diabetes, obesity and excess alcohol intake). However, much of the evidence comes from outside primary care, and it is doubtful whether those studies done in primary care used optimal intervention strategies. A priority for future research should be to demonstrate the feasibility, efficacy and efficiency of lifestyle interventions in a general practice setting. PMID:8731628

  7. Gonococcal interactions with polymorphonuclear neutrophils: importance of the phagosome for bactericidal activity.

    PubMed Central

    Densen, P; Mandell, G L

    1978-01-01

    Gonococci are capable of attaching to the surface of polymorphonuclear leukocytes (PMN). In this location they resist phagocytosis and are not killed by PMN. To delineate the factors involved in the survival of these gonococci, we investigated the interaction of virulent gonococci, which adhere to cells and resist phagocytosis, and avirulent gonococci, which are phagocytized and killed by PMN. In the presence of serum, both virulent and avirulent gonococci associate equally well with PMN and stimulate increases in oxidative metabolism. In the absence of serum virulent gonococci attached to PMN and stimulated PMN oxidative metabolism to a greater extent than avirulent gonococci which did not attach to PMN (P = 0.0009). Therefore, the survival of virulent gonococci attached to the PMN surface is not a result of failure to activate oxidative and bactericidal mechanisms. Both virulent and avirulent gonococci stimulated equivalent PMN specific granule release as measured by the appearance of lactoferrin in the media. Phagocytosis of avirulent gonococci stimulated significantly greater beta-glucuronidase release (P = 0.01) and myeloperoxidase-mediated iodination of protein (P = 0.001) by PMN than attachment of virulent gonococci. In the absence of serum neither type of gonococci stimulated beta-glocuronidase release or protein iodination by PMN. Thus, virulent gonococci fail to stimulate primary granule release by PMN. To further assess the role of attachment versus ingestion on the survival of gonococci, PMN were treated with cytochalasin B to block ingestion. Cytochalasin B-treated PMN were unable to kill either virulent or avirulent gonococci despite normal degranulation stimulated by the latter. The failure of PMN to kill surface-attached gonococci appears to be a consequence of the failure of PMN to enclose the virulent gonococci within a phagosome. The phagocytic vacuole thus plays a critical role in normal PMN bactericidal activity by providing a closed space in

  8. Are Parental Perceptions of Child Activity Levels and Overall Health More Important than Perceptions of Weight?

    PubMed Central

    Vangeepuram, Nita; Ramos, Michelle A.; Fei, Kezhen; Fox, Ashley M.; Horowitz, Carol R.; Kleinman, Lawrence C.; Galvez, Maida P.

    2016-01-01

    Objectives To examine relationships between parental perceptions of child weight and overall health, reported lifestyle behaviors and measured body mass index (BMI). Methods Using community-partnered methods, we surveyed families residing in a two census tract area identified for targeted interventions to decrease diabetes related disparities. The survey included demographics, child dietary and physical activity behaviors, and parental perception of child’s health and weight. We measured child BMI using a standardized protocol. Results We surveyed parents of 116 children with a mean age of 7 years (range 3–15) with 51 % boys, 74 % Hispanic, and 26 % Black. Over half of the children (55 %) were overweight or obese. Half (50 %) of the parents underestimated their children’s weight. Reported daily hours of walking and/or running trended higher (3.6 vs. 2.6 h, p = 0.08) for children perceived to be of normal weight. Parents who correctly estimated their child’s weight status reported more hours of daily walking/running than parents who underestimated child weight status, 4.5 versus 2.4 h, p = 0.0002. Parents of healthy weight children were more likely to report that children were in excellent or very good health compared to parents of overweight/obese children, 75 versus 56 % respectively (p = 0.04). We found significant racial/ethnic differences in reported diet and physical activity behaviors and perception of overall health. Conclusions for Practice Parental perceptions of child health and physical activity level may be related to perceptions of their child’s weight status. Study findings informed community-based initiatives for reducing diabetes risk among children. PMID:27010551

  9. Comparative activities of clavulanic acid, sulbactam, and tazobactam against clinically important beta-lactamases.

    PubMed Central

    Payne, D J; Cramp, R; Winstanley, D J; Knowles, D J

    1994-01-01

    Clavulanic acid, sulbactam, and tazobactam are inhibitors of a variety of plasmid-mediated beta-lactamases. However, inhibition data for these three inhibitors with a wide range of different plasmid-mediated beta-lactamases have not yet been compared under the same experimental conditions. A number of groups have inferred that clavulanic acid inhibits extended-spectrum TEM and SHV beta-lactamases, but inhibition data have rarely been published. In this study, the 50% inhibitory concentrations of these three beta-lactamase inhibitors for 35 plasmid-mediated beta-lactamases have been determined. Of these 35 beta-lactamases, 20 were extended-spectrum TEM- or SHV-derived beta-lactamases. The other 15 enzymes were conventional-spectrum beta-lactamases such as TEM-1 and SHV-1. Clavulanic acid was a more potent inhibitor than sulbactam for 32 of the 35 plasmid-mediated beta-lactamases tested. In particular, clavulanic acid was 60 and 580 times more potent than sulbactam against TEM-1 and SHV-1, respectively, currently the two most clinically prevalent gram-negative plasmid-mediated beta-lactamases. Statistical analysis of the data of the 50% inhibitory concentrations showed that clavulanic acid was 20 times more active overall than sulbactam against the conventional-spectrum enzymes. In addition, clavulanic acid was 14 times more potent than sulbactam at inhibiting the extended-spectrum enzymes. Tazobactam also showed significantly greater activity than sulbactam against the two groups of beta-lactamases. There were no significant differences between the overall activities of tazobactam and clavulanic acid against the extended-spectrum TEM and SHV enzymes and conventional-spectrum enzymes, although differences in their inhibition profiles were observed. PMID:8031044

  10. Post-Transition State Dynamics in Gas Phase Reactivity: Importance of Bifurcations and Rotational Activation.

    PubMed

    Martín-Sómer, Ana; Yáñez, Manuel; Hase, William L; Gaigeot, Marie-Pierre; Spezia, Riccardo

    2016-03-01

    Beyond the established use of thermodynamic vs kinetic control to explain chemical reaction selectivity, the concept of bifurcations on a potential energy surface (PES) is proving to be of pivotal importance with regard to selectivity. In this article, we studied by means of post-transition state (TS) direct dynamics simulations the effect that vibrational and rotational excitation at the TS may have on selectivity on a bifurcating PES. With this aim, we studied the post-TS unimolecular reactivity of the [Ca(formamide)](2+) ion, for which Coulomb explosion and neutral loss reactions compete. The PES exhibits different kinds of nonintrinsic reaction coordinate (IRC) dynamics, among them PES bifurcations, which direct the trajectories to multiple reaction paths after passing the TS. Direct dynamics simulations were used to distinguish between the bifurcation non-IRC dynamics and non-IRC dynamics arising from atomistic motions directing the trajectories away from the IRC. Overall, we corroborated the idea that kinetic selectivity often does not reduce to a simple choice between paths with different barrier heights and instead dynamical behavior after passing the TS may be crucial. Importantly, rotational excitation may play a pivotal role on the reaction selectivity favoring nonthermodynamic products.

  11. Retinobenzoic acids. 4. Conformation of aromatic amides with retinoidal activity. Importance of trans-amide structure for the activity.

    PubMed

    Kagechika, H; Himi, T; Kawachi, E; Shudo, K

    1989-10-01

    N-Methylation of two retinoidal amide compounds, 4-[(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthalenyl)carbamoyl]benz oic acid (3, Am80) and 4-[[(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2- naphthalenyl)carbonyl]amino]benzoic acid (5, Am580), resulted in the disappearance of their potent differentiation-inducing activity on human promyelocytic leukemia cell line HL-60. Studies with 1H NMR and UV spectroscopy indicated that large conformational differences exist between the active secondary amides and the inactive N-methyl amides. From a comparison of the spectroscopic results of these amides with those of stilbene derivatives, the conformations of the active amides are expected to resemble that of (E)-stilbene, whereas the inactive amides resemble the Z isomer: 3 (Am80) and 5 (Am580) have a trans-amide bond and their whole structures are elongated, while the N-methylated compounds [4 (Am90) and 6 (Am590)] have a cis-amide bond, resulting in the folding of the two benzene rings. These structures in the crystals were related to those in solution by 13C NMR spectroscopic comparison between the two phases (solid and solution).

  12. Covalent Inhibition of Ubc13 Affects Ubiquitin Signaling and Reveals Active Site Elements Important for Targeting.

    PubMed

    Hodge, Curtis D; Edwards, Ross A; Markin, Craig J; McDonald, Darin; Pulvino, Mary; Huen, Michael S Y; Zhao, Jiyong; Spyracopoulos, Leo; Hendzel, Michael J; Glover, J N Mark

    2015-07-17

    Ubc13 is an E2 ubiquitin conjugating enzyme that functions in nuclear DNA damage signaling and cytoplasmic NF-κB signaling. Here, we present the structures of complexes of Ubc13 with two inhibitors, NSC697923 and BAY 11-7082, which inhibit DNA damage and NF-κB signaling in human cells. NSC697923 and BAY 11-7082 both inhibit Ubc13 by covalent adduct formation through a Michael addition at the Ubc13 active site cysteine. The resulting adducts of both compounds exploit a binding groove unique to Ubc13. We developed a Ubc13 mutant which resists NSC697923 inhibition and, using this mutant, we show that the inhibition of cellular DNA damage and NF-κB signaling by NSC697923 is largely due to specific Ubc13 inhibition. We propose that unique structural features near the Ubc13 active site could provide a basis for the rational development and design of specific Ubc13 inhibitors. PMID:25909880

  13. Covalent Inhibition of Ubc13 Affects Ubiquitin Signaling and Reveals Active Site Elements Important for Targeting.

    PubMed

    Hodge, Curtis D; Edwards, Ross A; Markin, Craig J; McDonald, Darin; Pulvino, Mary; Huen, Michael S Y; Zhao, Jiyong; Spyracopoulos, Leo; Hendzel, Michael J; Glover, J N Mark

    2015-07-17

    Ubc13 is an E2 ubiquitin conjugating enzyme that functions in nuclear DNA damage signaling and cytoplasmic NF-κB signaling. Here, we present the structures of complexes of Ubc13 with two inhibitors, NSC697923 and BAY 11-7082, which inhibit DNA damage and NF-κB signaling in human cells. NSC697923 and BAY 11-7082 both inhibit Ubc13 by covalent adduct formation through a Michael addition at the Ubc13 active site cysteine. The resulting adducts of both compounds exploit a binding groove unique to Ubc13. We developed a Ubc13 mutant which resists NSC697923 inhibition and, using this mutant, we show that the inhibition of cellular DNA damage and NF-κB signaling by NSC697923 is largely due to specific Ubc13 inhibition. We propose that unique structural features near the Ubc13 active site could provide a basis for the rational development and design of specific Ubc13 inhibitors.

  14. Covalent Inhibition of Ubc13 Affects Ubiquitin Signaling and Reveals Active Site Elements Important for Targeting

    PubMed Central

    Hodge, Curtis D.; Edwards, Ross A.; Markin, Craig J.; McDonald, Darin; Pulvino, Mary; Huen, Michael S. Y.; Zhao, Jiyong; Spyracopoulos, Leo; Hendzel, Michael J.; Glover, J.N. Mark

    2015-01-01

    Ubc13 is an E2 ubiquitin conjugating enzyme that functions in nuclear DNA damage signaling and cytoplasmic NF-κB signaling. Here we present the structures of complexes of Ubc13 with two inhibitors, NSC697923 and BAY 11-7082, which inhibit DNA damage and NF-κB signaling in human cells. NSC697923 and BAY 11-7082 both inhibit Ubc13 by covalent adduct formation through a Michael addition at the Ubc13 active site cysteine. The resulting adducts of both compounds exploit a binding groove unique to Ubc13. We developed a Ubc13 mutant which resists NSC697923 inhibition and, using this mutant, we show that the inhibition of cellular DNA damage and NF-κB signaling by NSC697923 is largely due to specific Ubc13 inhibition. We propose that unique structural features near the Ubc13 active site could provide a basis for the rational development and design of specific Ubc13 inhibitors. PMID:25909880

  15. Imparting super hydro/oleophobic properties to cotton fabric by means of molecular and nanoparticles vapor deposition methods

    NASA Astrophysics Data System (ADS)

    Aminayi, Payam; Abidi, Noureddine

    2013-12-01

    Super hydro/oleophobic cotton fabrics were prepared using nanoparticle vapor deposition and molecular vapor deposition techniques. The surface was roughened by trimethylaluminum/water nanoparticles followed by functionalization with (tridecafluoro-1,1,2,2-tetrahydrooctyl)trichlorosilane. This process imparted unique hydro/oleophobic properties (contact angle >160°). In contrast to wet chemistry processes, the chemicals used and the thickness of the coatings produced by this method could be controlled precisely, which allowed for the minimization of waste generation while preserving the original properties of the fabric. Dynamic contact angles were measured and the fabrics that were coated using this technique showed low-contact-angle hysteresis properties. Scanning electron microscopy and universal attenuated total reflectance Fourier transform infrared spectroscopy confirmed the formation of a nanostructure-roughened surface. Drop dynamics such as the force of droplet movement, work of adhesion, and surface free energy were also calculated.

  16. Importance of aerosol composition and mixing state for cloud droplet activation in the high Arctic

    NASA Astrophysics Data System (ADS)

    Leck, C.; Svensson, E.

    2014-08-01

    Concentrations of cloud condensation nuclei (CCN) were measured throughout an expedition by icebreaker around the central Arctic Ocean, including a 3 week ice drift operation at 87° N, from 3 August to 9 September 2008. In agreement with previous observations in the area and season median daily CCN concentrations at 0.2% water vapor supersaturation were typically in the range of 15 to 30 cm-3, but concentrations varied by two to three orders of magnitude over the expedition and were occasionally below 1 cm-3. The CCN concentrations were highest near the ice edge and fell by a factor of three in the first 48 h of transport from the open sea into the pack ice region. For longer transport times they increased again indicating a local source over the pack ice, suggested to be polymer gels, via drops injected into the air by bubbles bursting on open leads. By assuming Köhler theory and simulating the cloud nucleation process using a Lagrangian adiabatic air parcel model that solves the kinetic formulation for condensation of water on size resolved aerosol particles we inferred the properties of the unexplained non-water soluble aerosol fraction that is necessary for reproducing the observed concentrations of CCN. We propose that the portion of the internally/externally mixed water insoluble particles was larger in the corresponding smaller aerosol sizes ranges. These particles were physically and chemically behaving as polymer gels: the interaction of the hydrophilic and hydrophobic entities on the structures of polymer gels during cloud droplet activation would at first only show a partial wetting character and only weak hygroscopic growth. Given time, a high CCN activation efficiency is achieved, which is promoted by the hydrophilicity or surface-active properties of the gels. Thus the result in this study argues for that the behavior of the high Arctic aerosol in CCN-counters operating at water vapor supersaturations > 0.4% (high relative humidities) may not be

  17. Is anterior cruciate ligament surgery technique important in rehabilitation and activity scores?

    PubMed Central

    Kilinc, Bekir Eray; Kara, Adnan; Celik, Haluk; Oc, Yunus; Camur, Savas

    2016-01-01

    To compare the two different anterior cruciate ligament surgery techniques’ effect in rehabilitation and activity performance. Fifty-five patients were evaluated. Twenty-seven patients with transtibial technique (TT), 28 with anatomic single-bundle technique (AT) included. Tegner Activity Scale (TAS) was performed at preoperation and follow-up. The returning time of the sport and work was evaluated at follow-up. Single-leg hop test was performed at follow-up. Outcomes were compared between the two groups. The determined length difference between the operated knee and the intact knee was compared between the two groups. Average age of TT and AT was 27.9±6.4 yr, 28.3±6 yr, respectively. There was a significant difference between the two groups in duration of returning to sport. TT group had higher duration to return to sport (P<0.01). No difference between the two groups in duration of returning to work (P>0.05). There was a significant difference between the two groups. TT group had significantly higher values than AT group (P<0.01). No difference in TAS between the two techniques at preoperation and at last follow-up (P>0.05). The increase of TAS in patients who had AT was higher than the patients who had TT (P>0.05). No difference in single-leg hop test at 55%–65%, 65%–75%, and 85%–95% level (P>0.05). In this test at 75%–85% TT group had higher values than AT group (P<0.05), AT group had higher values at 95%–105% level (P<0.05). Good short and long-term knee outcome scores depend on rehabilitation protocol after surgery. Surgery technique should provide the adequate stability in rehabilitation period. AT obtains better outcomes in rehabilitation. PMID:27419120

  18. Importance of lysine 125 for heparin binding and activation of antithrombin.

    PubMed

    Schedin-Weiss, Sophia; Desai, Umesh R; Bock, Susan C; Gettins, Peter G W; Olson, Steven T; Björk, Ingemar

    2002-04-16

    The anticoagulant sulfated polysaccharide, heparin, binds to the plasma coagulation proteinase inhibitor, antithrombin, and activates it by a conformational change that results in a greatly increased rate of inhibition of target proteinases. Lys125 of antithrombin has previously been implicated in this binding by chemical modification and site-directed mutagenesis and by the crystal structure of a complex between antithrombin and a pentasaccharide constituting the antithrombin-binding region of heparin. Replacement of Lys125 with Met or Gln in this work reduced the affinity of antithrombin for full-length heparin or the pentasaccharide by 150-600-fold at I = 0.15, corresponding to a loss of 25-33% of the total binding energy. The affinity decrease was due both to disruption of approximately three ionic interactions, indicating that Lys125 and two other basic residues of antithrombin act cooperatively in binding to heparin, and to weakened nonionic interactions. The mutations caused a 10-17-fold decrease in the affinity of the initial, weak binding step of the two-step mechanism of heparin binding to antithrombin. They also increased the reverse rate constant of the second, conformational change step by 10-50-fold. Lys125 is thus a major heparin-binding residue of antithrombin, contributing an amount of binding energy comparable to that of Arg129, but less energy than Lys114. It is the first residue identified so far that has a critical role in the initial recognition of heparin by antithrombin, but also appreciably stabilizes the heparin-induced activated state of the inhibitor. These effects are exerted by interactions of Lys125 with the nonreducing end of the heparin pentasaccharide. PMID:11939772

  19. Downregulation of hPMC2 imparts chemotherapeutic sensitivity to alkylating agents in breast cancer cells

    PubMed Central

    Krishnamurthy, Nirmala; Liu, Lili; Xiong, Xiahui; Zhang, Junran; Montano, Monica M

    2015-01-01

    Triple negative breast cancer cell lines have been reported to be resistant to the cyotoxic effects of temozolomide (TMZ). We have shown previously that a novel protein, human homolog of Xenopus gene which Prevents Mitotic Catastrophe (hPMC2) has a role in the repair of estrogen-induced abasic sites. Our present study provides evidence that downregulation of hPMC2 in MDA-MB-231 and MDA-MB-468 breast cancer cells treated with temozolomide (TMZ) decreases cell survival. This increased sensitivity to TMZ is associated with an increase in number of apurinic/apyrimidinic (AP) sites in the DNA. We also show that treatment with another alkylating agent, BCNU, results in an increase in AP sites and decrease in cell survival. Quantification of western blot analyses and immunofluorescence experiments reveal that treatment of hPMC2 downregulated cells with TMZ results in an increase in γ-H2AX levels, suggesting an increase in double strand DNA breaks. The enhancement of DNA double strand breaks in TMZ treated cells upon downregulation of hPCM2 is also revealed by the comet assay. Overall, we provide evidence that downregulation of hPMC2 in breast cancer cells increases cytotoxicity of alkylating agents, representing a novel mechanism of treatment for breast cancer. Our data thus has important clinical implications in the management of breast cancer and brings forth potentially new therapeutic strategies. PMID:25849309

  20. Imparting electroactivity to polycaprolactone fibers with heparin-doped polypyrrole: Modulation of hemocompatibility and inflammatory responses.

    PubMed

    Xiong, Gordon M; Yuan, Shaojun; Wang, Jun Kit; Do, Anh Tuan; Tan, Nguan Soon; Yeo, Kiat Seng; Choong, Cleo

    2015-09-01

    Hemocompatibility, anti-inflammation and anti-thrombogenicity of acellular synthetic vascular grafts remains a challenge in biomaterials design. Using electrospun polycaprolactone (PCL) fibers as a template, a coating of polypyrrole (PPy) was successfully polymerized onto the fiber surface. The fibers coated with heparin-doped PPy (PPy-HEP) demonstrated better electroactivity, lower surface resistivity (9-10-fold) and better anti-coagulation response (non-observable plasma recalcification after 30min vs. recalcification at 8-9min) as compared to fibers coated with pristine PPy. Red blood cell compatibility, measured by% hemolysis, was greatly improved on PPy-HEP-coated PCL in comparison to uncoated PCL (3.9±2.1% vs. 22.1±4.1%). PPy-HEP-coated PCL fibers also exhibited higher stiffness values (6.8±0.9MPa vs. 4.2±0.8MPa) as compared to PCL fibers, but similar tensile strengths. It was also observed that the application of a low alternating current led to a 4-fold reduction of platelet activation (as quantitated by CD62p expression) for the PPy-HEP-coated fibers as compared to non-stimulated conditions. In parallel, a reduction in the leukocyte adhesion to both pristine PPy-coated and PPy-HEP-coated fibers was observable with AC stimulation. Overall, a new strategy involving the use of hemocompatible conducting polymers and electrical stimulation to control thrombogenicity and inflammatory responses for synthetic vascular graft designs was demonstrated.

  1. Recombinant BCG Expressing Mycobacterium ulcerans Ag85A Imparts Enhanced Protection against Experimental Buruli ulcer

    PubMed Central

    Hart, Bryan E.; Hale, Laura P.; Lee, Sunhee

    2015-01-01

    Buruli ulcer, an emerging tropical disease caused by Mycobacterium ulcerans (MU), is characterized by disfiguring skin necrosis and high morbidity. Relatively little is understood about the mode of transmission, pathogenesis, or host immune responses to MU infection. Due to significant reduction in quality of life for patients with extensive tissue scarring, and that a disproportionately high percentage of those affected are disadvantaged children, a Buruli ulcer vaccine would be greatly beneficial to the worldwide community. Previous studies have shown that mice inoculated with either M. bovis bacille Calmette–Guérin (BCG) or a DNA vaccine encoding the M. ulcerans mycolyl transferase, Ag85A (MU-Ag85A), are transiently protected against pathology caused by intradermal challenge with MU. Building upon this principle, we have generated quality-controlled, live-recombinant strains of BCG and M. smegmatis which express the immunodominant MU Ag85A. Priming with rBCG MU-Ag85A followed by an M. smegmatis MU-Ag85A boost strongly induced murine antigen-specific CD4+ T cells and elicited functional IFNγ-producing splenocytes which recognized MU-Ag85A peptide and whole M. ulcerans better than a BCG prime-boost vaccination. Strikingly, mice vaccinated with a single subcutaneous dose of BCG MU-Ag85A or prime-boost displayed significantly enhanced survival, reduced tissue pathology, and lower bacterial load compared to mice vaccinated with BCG. Importantly, this level of superior protection against experimental Buruli ulcer compared to BCG has not previously been achieved. These results suggest that use of BCG as a recombinant vehicle expressing MU antigens represents an effective Buruli ulcer vaccine strategy and warrants further antigen discovery to improve vaccine efficacy. PMID:26393347

  2. Dissipation of the energy imparted by mid-latitude storms in the Southern Ocean

    NASA Astrophysics Data System (ADS)

    Jouanno, Julien; Capet, Xavier; Madec, Gurvan; Roullet, Guillaume; Klein, Patrice

    2016-06-01

    The aim of this study is to clarify the role of the Southern Ocean storms on interior mixing and meridional overturning circulation. A periodic and idealized numerical model has been designed to represent the key physical processes of a zonal portion of the Southern Ocean located between 70 and 40° S. It incorporates physical ingredients deemed essential for Southern Ocean functioning: rough topography, seasonally varying air-sea fluxes, and high-latitude storms with analytical form. The forcing strategy ensures that the time mean wind stress is the same between the different simulations, so the effect of the storms on the mean wind stress and resulting impacts on the Southern Ocean dynamics are not considered in this study. Level and distribution of mixing attributable to high-frequency winds are quantified and compared to those generated by eddy-topography interactions and dissipation of the balanced flow. Results suggest that (1) the synoptic atmospheric variability alone can generate the levels of mid-depth dissipation frequently observed in the Southern Ocean (10-10-10-9 W kg-1) and (2) the storms strengthen the overturning, primarily through enhanced mixing in the upper 300 m, whereas deeper mixing has a minor effect. The sensitivity of the results to horizontal resolution (20, 5, 2 and 1 km), vertical resolution and numerical choices is evaluated. Challenging issues concerning how numerical models are able to represent interior mixing forced by high-frequency winds are exposed and discussed, particularly in the context of the overturning circulation. Overall, submesoscale-permitting ocean modeling exhibits important delicacies owing to a lack of convergence of key components of its energetics even when reaching Δx = 1 km.

  3. The Effects of Constraint-Induced Movement Therapy on Activities Important to Independent School Participation of Children with Hemiparesis

    ERIC Educational Resources Information Center

    Carney, Joan

    2012-01-01

    This study investigated the efficacy of constraint-induced movement therapy (CI therapy) on activities important to school participation in children with hemiparesis. Four children, ages 4-0 to 7-10 participated in an intensive CI therapy program in a clinical setting. Constraining casts were worn 24 hours daily. Therapy was delivered 6 hours…

  4. Imparting electroactivity to polycaprolactone fibers with heparin-doped polypyrrole: Modulation of hemocompatibility and inflammatory responses.

    PubMed

    Xiong, Gordon M; Yuan, Shaojun; Wang, Jun Kit; Do, Anh Tuan; Tan, Nguan Soon; Yeo, Kiat Seng; Choong, Cleo

    2015-09-01

    Hemocompatibility, anti-inflammation and anti-thrombogenicity of acellular synthetic vascular grafts remains a challenge in biomaterials design. Using electrospun polycaprolactone (PCL) fibers as a template, a coating of polypyrrole (PPy) was successfully polymerized onto the fiber surface. The fibers coated with heparin-doped PPy (PPy-HEP) demonstrated better electroactivity, lower surface resistivity (9-10-fold) and better anti-coagulation response (non-observable plasma recalcification after 30min vs. recalcification at 8-9min) as compared to fibers coated with pristine PPy. Red blood cell compatibility, measured by% hemolysis, was greatly improved on PPy-HEP-coated PCL in comparison to uncoated PCL (3.9±2.1% vs. 22.1±4.1%). PPy-HEP-coated PCL fibers also exhibited higher stiffness values (6.8±0.9MPa vs. 4.2±0.8MPa) as compared to PCL fibers, but similar tensile strengths. It was also observed that the application of a low alternating current led to a 4-fold reduction of platelet activation (as quantitated by CD62p expression) for the PPy-HEP-coated fibers as compared to non-stimulated conditions. In parallel, a reduction in the leukocyte adhesion to both pristine PPy-coated and PPy-HEP-coated fibers was observable with AC stimulation. Overall, a new strategy involving the use of hemocompatible conducting polymers and electrical stimulation to control thrombogenicity and inflammatory responses for synthetic vascular graft designs was demonstrated. PMID:25983317

  5. Imparting chemical stability in nanoparticulate silver via a conjugated polymer casing approach.

    PubMed

    Chang, Mincheol; Kim, Taejoon; Park, Hyun-Woo; Kang, Minjeong; Reichmanis, Elsa; Yoon, Hyeonseok

    2012-08-01

    Only limited information is available on the design and synthesis of functional materials for preventing corrosion of metal nanostructures. In the nanometer regime, even noble metals are subject to chemical attack. Here, the corrosion behavior of noble metal nanoparticles coated with a conjugated polymer nanolayer was explored for the first time. Specifically, electrochemical corrosion and sulfur tarnishing behaviors were examined for Ag-polypyrrole (PPy) core-shell nanoparticles using potentiodynamic polarization and spectrophotometric analysis, respectively. First, the Ag-PPy nanoparticles exhibited enhanced resistance to electrochemically induced corrosion compared to their exposed silver counterparts. Briefly, a neutral PPy shell provided the highest protection efficiency (75.5%), followed by sulfate ion- (61.3%) and dodecylbenzenesulfonate ion- (53.6%) doped PPy shells. However, the doping of the PPy shell with chloride ion induced an adverse effect (protection efficiency, -120%). Second, upon exposure to sulfide ions, the Ag-PPy nanoparticles preserved their morphology and colloidal stability while the bare silver analog underwent significant structural deformation. To further understand the function of the PPy shell as a protection layer for the silver core, the catalytic activity of the nanostructures was also evaluated. Using the reduction of 4-nitrophenol as a representative example of a catalytic reaction, the rate constant for that reduction using the PPy encased Ag nanoparticles was found to be 1.1 × 10(-3) s(-1), which is approximately 33% less than that determined for the parent silver. These results demonstrate that PPy can serve as both an electrical and chemical barrier for mitigating undesirable chemical degradation in corrosive environments, as well as provide a simple physical barrier to corrosive substances under appropriate conditions. PMID:22860984

  6. Trimethyloxonium modification of batrachotoxin-activated Na channels alters functionally important protein residues.

    PubMed Central

    Cherbavaz, D B

    1995-01-01

    The extracellular side of single batrachotoxin-activated voltage-dependent Na channels isolated from rat skeletal muscle membranes incorporated into neutral planar lipid bilayers were treated in situ with the carboxyl methylating reagent, trimethyloxonium (TMO). These experiments were designed to determine whether TMO alters Na channel function by a general through-space electrostatic mechanism or by methylating specific carboxyl groups essential to channel function. TMO modification reduced single-channel conductance by decreasing the maximal turnover rate. Modification increased channel selectivity for sodium ions relative to potassium ions as measured under biionic conditions. TMO modification increased the mu-conotoxin (muCTX) off-rate by three orders of magnitude. Modification did not alter the muCTX on-rate at low ionic strength or Na channel voltage-dependent gating characteristics. These data demonstrate that TMO does not act via a general electrostatic mechanism. Instead, TMO targets protein residues specifically involved in ion conduction, ion selectivity, and muCTX binding. These data support the hypothesis that muCTX blocks open-channel current by physically obstructing the ion channel pore. PMID:7787022

  7. The importance of the active surface area of graphite materials in the first lithium intercalation

    NASA Astrophysics Data System (ADS)

    Novák, P.; Ufheil, J.; Buqa, H.; Krumeich, F.; Spahr, M. E.; Goers, D.; Wilhelm, H.; Dentzer, J.; Gadiou, R.; Vix-Guterl, C.

    When lithium is intercalated into graphite in ethylene carbonate (EC) containing electrolytes, solvent co-intercalation leading to the exfoliation of the graphite structure could occur. The exfoliation can be suppressed if an efficient solid electrolyte interphase (SEI, a passivation layer) is formed. Here we study the role played by the active surface area (ASA) of graphite materials during their first electrochemical reduction. ASA (related to the presence of defects at the carbon surface) appears as a critical graphite surface parameter influencing the surface passivation mechanism and the graphite exfoliation. The ASA of TIMREX ® SLX50 synthetic graphite was modified by thermal treatment in argon and air. The electrochemical performance was characterized in 1 M LiPF 6, EC:DMC electrolyte and post mortem analyses were performed by SEM imaging. It turned out that a decrease of the graphite ASA, i.e., an increase of the graphite structural order, hinders the formation of the passivation layer and favors the exfoliation process. In contrast, the exfoliation of the same graphite can be suppressed if its ASA is increased for example by air treatment. The ASA of the graphite kinetically controls the formation of an efficient SEI film and accordingly the irreversible charge loss is much lower in the case of graphite with a high ASA value.

  8. Active processes on a mixed clastic carbonate Brazilian shelf margin: Importance for hydrocarbon exploration in turbidites

    SciTech Connect

    Cainelli, C. )

    1991-03-01

    The search for subtle hydrocarbon accumulations in turbidite systems requires additional approaches for more successful exploration, particularly when direct recognition on seismic lines is difficult. This includes the determination and understanding of processes controlling sand distribution on the shelf and the mapping of sand pathways from the shelf to the slop/basin that can guide efforts to look for more favorable sites for turbidite sandstone deposition. The approach can be exemplified in the Sergipe-Alagoas basin, on the Brazillian Atlantic passive margin. The section analyzed is the Piacabucu Formation, a thick seaward prograding wedge composed of coastal sandstones and shelf edge carbonates on a narrow shelf and slope-basin shales with turbidite lenses. Waves and currents control the redistribution of sediments transported to the shelf by rivers. More wave energy is expended in ten hours in the San Francisco delta than in an entire year in the Mississippi delta. Such environment precludes deposition of mud on the shelf, but it stimulates the development of shelf edge carbonates. Rimed carbonates along the shelf break serve as a barrier for downslope movements of coarse-grained sediment, where turbidites are oil targets. The search for gaps in the carbonate barrier which can tap the behind-barrier sands is critical for sand-rich turbidite development. It is believed that canyons create these gaps and act as active turbidity current routes.

  9. The importance of hinge sequence for loop function and catalytic activity in the reaction catalyzed by triosephosphate isomerase.

    PubMed

    Xiang, J; Sun, J; Sampson, N S

    2001-04-01

    We have determined the sequence requirements for the N-terminal protein hinge of the active-site lid of triosephosphate isomerase. The codons for the hinge (PVW) were replaced with a genetic library of all possible 8000 amino acid combinations. The most active of these 8000 mutants were selected using in vivo complementation of a triosephosphate isomerase-deficient strain of Escherichia coli, DF502. Approximately 0.3 % of the mutants complement DF502 with an activity that is between 10 and 70 % of wild-type activity. They all contain Pro at the first position. Furthermore, the sequences of these hinge mutants reveal that hydrophobic packing is very important for efficient formation of the enediol intermediate. However, the reduced catalytic activities observed are not due to increased rates of loop opening. To explore the relationship between the N-terminal and C-terminal hinges, three semi-active mutants from the N-terminal hinge selection experiment (PLH, PHS and PTF), and six active C-terminal hinge mutants from previous work (NSS, LWA, YSL, KTK, NPN, KVA) were combined to form 18 "double-hinge" mutants. The activities of these mutants suggest that the N-terminal and C-terminal hinge structures affect one another. It appears that specific side-chain interactions are important for forming a catalytically active enzyme, but not for preventing release of the unstable enediol intermediate from the active site of the enzyme. The independence of intermediate release on amino acid sequence is consistent with the absence of a "universal" hinge sequence in structurally related enzymes. PMID:11286559

  10. The importance of hinge sequence for loop function and catalytic activity in the reaction catalyzed by triosephosphate isomerase.

    PubMed

    Xiang, J; Sun, J; Sampson, N S

    2001-04-01

    We have determined the sequence requirements for the N-terminal protein hinge of the active-site lid of triosephosphate isomerase. The codons for the hinge (PVW) were replaced with a genetic library of all possible 8000 amino acid combinations. The most active of these 8000 mutants were selected using in vivo complementation of a triosephosphate isomerase-deficient strain of Escherichia coli, DF502. Approximately 0.3 % of the mutants complement DF502 with an activity that is between 10 and 70 % of wild-type activity. They all contain Pro at the first position. Furthermore, the sequences of these hinge mutants reveal that hydrophobic packing is very important for efficient formation of the enediol intermediate. However, the reduced catalytic activities observed are not due to increased rates of loop opening. To explore the relationship between the N-terminal and C-terminal hinges, three semi-active mutants from the N-terminal hinge selection experiment (PLH, PHS and PTF), and six active C-terminal hinge mutants from previous work (NSS, LWA, YSL, KTK, NPN, KVA) were combined to form 18 "double-hinge" mutants. The activities of these mutants suggest that the N-terminal and C-terminal hinge structures affect one another. It appears that specific side-chain interactions are important for forming a catalytically active enzyme, but not for preventing release of the unstable enediol intermediate from the active site of the enzyme. The independence of intermediate release on amino acid sequence is consistent with the absence of a "universal" hinge sequence in structurally related enzymes.

  11. {beta}-Catenin stabilization imparts crypt progenitor phenotype to hyperproliferating colonic epithelia

    SciTech Connect

    Sellin, Joseph H.; Wang Yu; Singh, Pomila; Umar, Shahid

    2009-01-01

    Utilizing the Citrobacter rodentium (CR)-induced transmissible murine colonic hyperplasia (TMCH) model, we provide mechanistic basis of changes in {beta}-catenin/APC/CKI{epsilon} leading to progression and/or regression of hyperplasia in vivo. In response to CR-induced TMCH, crypt lengths increased significantly between days 6-27 post-infection, followed by a steep decline by day 34. {beta}-Cat{sup 45}/total {beta}-catenin were elevated on day 1 post-infection, preceding changes in crypt length, and persisted for 27 days before declining by day 34. Importantly, cellular CKI{epsilon} and {beta}-catenin co-immunoprecipitated and exhibited remarkable parallel changes in kinetics during hyperplasia/regression phases. {beta}-catenin, phosphorylated at Ser33,37 and Thr41 ({beta}-cat{sup 33,37/41}), was low till day 12, followed by gradual increase until day 27 before declining by day 34. GSK-3{beta} exhibited significant Ser{sup 9}-phosphorylation/inactivation at days 6-12 with partial recovery at days 27-34. Wild type (wt) APC (p312) levels increased at day 6 with transient proteolysis/truncation to p130 form between days 12 and 15; p312 reappeared by day 19 and returned to baseline by day 34. The kinetics of {beta}-Cat{sup 45}/{beta}-catenin nuclear accumulation and acetylation (Ac-{beta}-Cat{sup Lys49}) from days 6 to 27, followed by loss of phosphorylation/acetylation by day 34 was almost identical; Tcf-4 co-immunoprecipitated with {beta}-Cat{sup 45}/{beta}-catenin and localized immunohistochemically to {beta}-Cat{sup 41/45}-positive regions leading to elevated cyclin D1 expression, during the hyperproliferative, but not regression phases of TMCH. CKI{epsilon} mediated phosphorylation of {beta}-Cat{sup 45}, resulting in stabilization/nuclear translocation of {beta}-Cat{sup 45} may be critical for maintaining proliferation at days 6-27. Reversal of GSK-3{beta} phosphorylation and APC changes may be equally critical during the regression phase from days 27 to 34.

  12. The importance of methanotrophic activity in geothermal soils of Pantelleria island (Italy)

    NASA Astrophysics Data System (ADS)

    D'Alessandro, Walter; Gagliano, Antonina Lisa; Quatrini, Paola; Parello, Francesco

    2013-04-01

    Methane is a major contributor to the greenhouse effect, its atmospheric concentration being more than doubled since the XIX century. Every year 22 Tg of methane are released to the atmosphere from several natural and anthropogenic sources. Natural sources include geothermal/volcanic areas but the estimation of the total methane emission from these areas is currently not well defined since the balance between emission through degassing and microbial oxidation within the soils is not well known. Microbial oxidation in soils contributes globally for about 3-9% to the removal of methane from the atmosphere and recent studies evidenced methanotrophic activity also in soils of volcanic/geothermal areas despite their harsh environmental conditions (high temperatures, low pH and high concentrations of H2S and NH3). Methanotrophs are a diverse group of bacteria that are able to metabolize methane as their only source of carbon and energy and are found within the Alpha and Gamma classes of Proteobacteria and within the phylum Verrucomicrobia. Our purpose was to study the interaction between methanotrophic communities and the methane emitted from the geothermally most active site of Pantelleria island (Italy), Favara Grande, whose total methane emission has been previously estimated in about 2.5 t/a. Laboratory incubation experiments with soil samples from Favara Grande showed methane consumption values of up to 9500 ng g-1 dry soil per hour while soils collected outside the geothermal area consume less than 6 ng g-1 h-1. The maximum consumption was measured in the shallowest part of the soil profile (1-3 cm) and high values (>100 ng g-1 h-1) were maintained up to a depht of 15 cm. Furthermore, the highest consumption was measured at 37°C, and a still recognizable consumption (>20 ng g-1 h-1) at 80°C, with positive correlation with the methane concentration in the incubation atmosphere. These results can be considered a clear evidence of the presence of methanotrophs that

  13. Importance of Baseline Prognostic Factors With Increasing Time Since Initiation of Highly Active Antiretroviral Therapy

    PubMed Central

    2012-01-01

    Background The extent to which the prognosis for AIDS and death of patients initiating highly active antiretroviral therapy (HAART) continues to be affected by their characteristics at the time of initiation (baseline) is unclear. Methods We analyzed data on 20,379 treatment-naive HIV-1–infected adults who started HAART in 1 of 12 cohort studies in Europe and North America (61,798 person-years of follow-up, 1844 AIDS events, and 1005 deaths). Results Although baseline CD4 cell count became less prognostic with time, individuals with a baseline CD4 count <25 cells/µL had persistently higher progression rates than individuals with a baseline CD4 count >350 cells/µL (hazard ratio for AIDS = 2.3, 95% confidence interval [CI]: 1.0 to 2.3; mortality hazard ratio = 2.5, 95% CI: 1.2 to 5.5, 4 to 6 years after starting HAART). Rates of AIDS were persistently higher in individuals who had experienced an AIDS event before starting HAART. Individuals with presumed transmission by means of injection drug use experienced substantially higher rates of AIDS and death than other individuals throughout follow-up (AIDS hazard ratio = 1.6, 95% CI: 0.8 to 3.0; mortality hazard ratio = 3.5, 95% CI: 2.2 to 5.5, 4 to 6 years after starting HAART). Conclusions Compared with other patient groups, injection drug users and patients with advanced immunodeficiency at baseline experience substantially increased rates of AIDS and death up to 6 years after starting HAART. PMID:18043315

  14. β-Catenin is important for cancer stem cell generation and tumorigenic activity in nasopharyngeal carcinoma.

    PubMed

    Jiang, Rui; Niu, Xiaoshuang; Huang, Yuxiang; Wang, Xiaosheng

    2016-03-01

    Nasopharyngeal carcinoma (NPC) is one of the most common malignant tumors with poor prognosis and recurrence in South China. The hard eradication of NPC in clinic is predominantly due to cancer stem cells (CSCs). Increasing evidence revealed that the aberrant activation of Wnt/β-catenin was positively correlated with the produce of CSCs. To further investigate the effect of β-catenin on CSCs and tumorigenesis in NPC, a CNE2 cell line (pLKO.1-sh-β-catenin-CNE2) with stably suppressed expression of β-catenin was used in this study. The expressions of biomarkers in CSCs including c-myc, Nanog, Oct3/4, Sox2, EpCAM as well as adhesion-related proteins like E-cadherin and vimentin were analyzed by western blot analysis and immunofluorescent staining. The proliferation and migration abilities were investigated by MTT assay and Transwell assay, respectively. Cell cycle was analyzed by flow cytometry. Finally, xenograft was performed to determine the effect of β-catenin on oncogenesis in vivo. Results showed that the expressions of c-myc, Nanog, Oct3/4, Sox2, and EpCAM were all decreased in pLKO.1-sh-β-catenin-CNE2 cells. It was also found that vimentin was downregulated, while E-cadherin was upregulated. Results of MTT and Transwell assays suggested that the proliferation and migration abilities were impaired by silencing of β-catenin, and more cells were arrested in G1 phase when compared with the control. In vivo study indicated that the tumor growth was markedly suppressed in experimental group. Based on current findings, β-catenin may function as an essential protein for the maintenance of migration and proliferation abilities of NPC cells, and a complicated network consisting of c-myc, Nanog, Oct3/4, Sox2, EpCAM, E-cadherin, vimentin, and β-catenin may be involved in the inherent regulation mechanisms. PMID:26849897

  15. Arginine kinase: differentiation of gene expression and protein activity in the red imported fire ant, Solenopsis invicta.

    PubMed

    Wang, Haichuan; Zhang, Lan; Zhang, Lee; Lin, Qin; Liu, Nannan

    2009-02-01

    Arginine kinase (AK), a primary enzyme in cell metabolism and adenosine 5'-triphosphate (ATP)-consuming processes, plays an important role in cellular energy metabolism and maintaining constant ATP levels in invertebrate cells. In order to identify genes that are differentially expressed between larvae and adults, queens and workers, and female alates (winged) and queens (wingless), AK cDNA was obtained from the red imported fire ant. The cDNA sequence of the gene has open reading frames of 1065 nucleotides, encoding a protein of 355 amino acid residues that includes the substrate recognition region, the signature sequence pattern of ATP:guanidino kinases, and an "actinin-type" actin binding domain. Northern blot analysis and protein activity analysis demonstrated that the expression of the AK gene and its protein activity were developmentally, caste specifically, and tissue specifically regulated in red imported fire ants with a descending order of worker> alate (winged adult) female> alate (winged adult) male> larvae> worker pupae approximately alate pupae. These results suggest a different demand for energy-consumption and production in the different castes of the red imported fire ant, which may be linked to their different missions and physiological activities in the colonies. The highest level of the AK gene expression and activity was identified in head tissue of both female alates and workers and thorax tissue of workers, followed by thorax tissue of female alates and abdomen tissue of male alates, suggesting the main tissues or cells in these body parts, such as brain, neurons and muscles, which have been identified as the major tissues and/or cells that display high and variable rates of energy turnover in other organisms, play a key role in energy production and its utilization in the fire ant. In contrast, in the male alate, the highest AK expression and activity were found in the abdomen, suggesting that here energy demand may relate to sperm formation

  16. Identification by mutagenesis of a conserved glutamate (Glu487) residue important for catalytic activity in rat liver carnitine palmitoyltransferase II.

    PubMed

    Zheng, Guolu; Dai, Jia; Woldegiorgis, Gebre

    2002-11-01

    Mammalian mitochondrial membranes express two active but distinct carnitine palmitoyltransferases: carnitine palmitoyltransferase I (CPTI), which is malonyl coA-sensitive and detergent-labile; and carnitine palmitoyltransferase II (CPTII), which is malonyl coA-insensitive and detergent-stable. To determine the role of the highly conserved C-terminal acidic residues glutamate 487 (Glu(487)) and glutamate 500 (Glu(500)) on catalytic activity in rat liver CPTII, we separately mutated these residues to alanine, aspartate, or lysine, and the effect of the mutations on CPTII activity was determined in the Escherichia coli-expressed mutants. Substitution of Glu(487) with alanine, aspartate, or lysine resulted in almost complete loss in CPTII activity. Because a conservative substitution mutation of this residue, Glu(487) with aspartate (E487D), resulted in a 97% loss in activity, we predicted that Glu(487) would be at the active-site pocket of CPTII. The substantial loss in CPTII activity observed with the E487K mutant, along with the previously reported loss in activity observed in a child with a CPTII deficiency disease, establishes that Glu(487) is crucial for maintaining the configuration of the liver isoform of the CPTII active site. Substitution of the conserved Glu(500) in CPTII with alanine or aspartate reduced the V(max) for both substrates, suggesting that Glu(500) may be important in stabilization of the enzyme-substrate complex. A conservative substitution of Glu(500) to aspartate resulted in a significant decrease in the V(max) for the substrates. Thus, Glu(500) may play a role in substrate binding and catalysis. Our site-directed mutagenesis studies demonstrate that Glu(487) in the liver isoform of CPTII is essential for catalysis. PMID:12200419

  17. The p21-activated kinase, PAK2, is important in the activation of numerous pancreatic acinar cell signaling cascades and in the onset of early pancreatitis events.

    PubMed

    Nuche-Berenguer, Bernardo; Ramos-Álvarez, Irene; Jensen, R T

    2016-06-01

    In a recent study we explored Group-1-p21-activated kinases (GP.1-PAKs) in rat pancreatic acini. Only PAK2 was present; it was activated by gastrointestinal-hormones/neurotransmitters and growth factors in a PKC-, Src- and small-GTPase-mediated manner. PAK2 was required for enzyme-secretion and ERK/1-2-activation. In the present study we examined PAK2's role in CCK and TPA-activation of important distal signaling cascades mediating their physiological/pathophysiological effects and analyzed its role in pathophysiological processes important in early pancreatitis. In rat pancreatic acini, PAK2-inhibition by the specific, GP.1.PAK-inhibitor, IPA-3-suppressed cholecystokinin (CCK)/TPA-stimulated activation of focal-adhesion kinases and mitogen-activated protein-kinases. PAK2-inhibition reversed the dual stimulatory/inhibitory effect of CCK/TPA on the PI3K/Akt/GSK-3β pathway. However, its inhibition did not affect PKC activation. PAK2-inhibition protected acini from CCK-induced ROS-generation; caspase/trypsin-activation, important in early pancreatitis; as well as from cell-necrosis. Furthermore, PAK2-inhibition reduced proteolytic-activation of PAK-2p34, which is involved in programmed-cell-death. To ensure that the study did not only rely in the specificity of IPA-3 as a PAK inhibitor, we used two other approaches for PAK inhibition, FRAX597 a ATP-competitive-GP.1-PAKs-inhibitor and infection with a PAK2-dominant negative(DN)-Advirus. Those two approaches confirmed the results obtained with IPA-3. This study demonstrates that PAK2 is important in mediating CCK's effect on the activation of signaling-pathways known to mediate its physiological/pathophysiological responses including several cellular processes linked to the onset of pancreatitis. Our results suggest that PAK2 could be a new, important therapeutic target to consider for the treatment of diseases involving deregulation of pancreatic acinar cells. PMID:26912410

  18. Synergistic Behavior of Tubes, Junctions, and Sheets Imparts Mechano-Mutable Functionality in 3D Porous Boron Nitride Nanostructures

    PubMed Central

    2015-01-01

    One-dimensional (1D) boron nitride nanotube (BNNT) and 2D hexagonal BN (h-BN) are attractive for demonstrating fundamental physics and promising applications in nano-/microscale devices. However, there is a high anisotropy associated with these BN allotropes as their excellent properties are either along the tube axis or in-plane directions, posing an obstacle in their widespread use in technological and industrial applications. Herein, we report a series of 3D BN prototypes, namely, pillared boron nitride (PBN), by fusing single-wall BNNT and monolayer h-BN aimed at filling this gap. We use density functional theory and molecular dynamics simulations to probe the diverse mechano-mutable properties of PBN prototypes. Our results demonstrate that the synergistic effect of the tubes, junctions, and sheets imparts cooperative deformation mechanisms, which overcome the intrinsic limitations of the PBN constituents and provide a number of superior characteristics including 3D balance of strength and toughness, emergence of negative Poisson’s ratio, and elimination of strain softening along the armchair orientation. These features, combined with the ultrahigh surface area and lightweight structure, render PBN as a 3D multifunctional template for applications in graphene-based nanoelectronics, optoelectronics, gas storage, and functional composites with fascinating in-plane and out-of-plane tailorable properties. PMID:25289114

  19. Lipocalin 2 Imparts Selective Pressure on Bacterial Growth in the Bladder and Is Elevated in Women with Urinary Tract Infection

    PubMed Central

    Steigedal, Magnus; Marstad, Anne; Haug, Markus; Damås, Jan K.; Strong, Roland K.; Roberts, Pacita L.; Himpsl, Stephanie D.; Stapleton, Ann; Hooton, Thomas M.; Mobley, Harry L. T.; Hawn, Thomas R.

    2014-01-01

    Competition for iron is a critical component of successful bacterial infections, but the underlying in vivo mechanisms are poorly understood. We have previously demonstrated that lipocalin 2 (LCN2) is an innate immunity protein that binds to bacterial siderophores and starves them for iron, thus representing a novel host defense mechanism to infection. In the present study we show that LCN2 is secreted by the urinary tract mucosa and protects against urinary tract infection (UTI). We found that LCN2 was expressed in the bladder, ureters, and kidneys of mice subject to UTI. LCN2 was protective with higher bacterial numbers retrieved from bladders of Lcn2-deficient mice than from wild-type mice infected with the LCN2-sensitive Escherichia coli strain H9049. Uropathogenic E. coli mutants in siderophore receptors for salmochelin, aerobactin, or yersiniabactin displayed reduced fitness in wild-type mice, but not in mice deficient of LCN2, demonstrating that LCN2 imparts a selective pressure on bacterial growth in the bladder. In a human cohort of women with recurrent E. coli UTIs, urine LCN2 levels were associated with UTI episodes and with levels of bacteriuria. The number of siderophore systems was associated with increasing bacteriuria during cystitis. Our data demonstrate that LCN2 is secreted by the urinary tract mucosa in response to uropathogenic E. coli challenge and acts in innate immune defenses as a colonization barrier that pathogens must overcome to establish infection. PMID:25398327

  20. Importance of fatty acid substituents of chemically synthesized lipid A-subunit analogs in the expression of immunopharmacological activity.

    PubMed Central

    Kumazawa, Y; Nakatsuka, M; Takimoto, H; Furuya, T; Nagumo, T; Yamamoto, A; Homma, Y; Inada, K; Yoshida, M; Kiso, M

    1988-01-01

    The immunopharmacological activities of chemically synthesized lipid A-subunit analogs, 4-O-phosphono-D-glucosamine derivatives carrying different N- and 3-O-linked acyl groups, were investigated. None of the synthetic compounds tested exhibited any detectable pyrogenicity at a dose of 10 micrograms/kg. Weaker lethal toxicity in galactosamine-sensitized mice was detected at 1 microgram per mouse for all the synthetic compounds except GLA-58. Among (RS) stereoisomers of 4-O-phosphono-D-glucosamine derivatives carrying a 3-O-tetradecanoyl (C14) group with different N-linked acyloxyacyl groups, i.e., 3-dodecanoyloxytetradecanoyl [C14-O-(C12)], 3-tetradecanoyloxytetradecanoyl [C14-O-(C14)], and 3-hexadecanoyloxytetradecanoyl [C14-O-(C16)] groups (termed GLA-57, GLA-27, and GLA-58, respectively), GLA-27 exhibited significant colony-stimulating factor-inducing and tumor necrosis factor-inducing activities, mitogenicity, polyclonal B-cell activation activity, macrophage activation, and adjuvanticity. The activities of GLA-57, which had an N-linked C14-O-(C12) group, were equivalent to or somewhat weaker than those of GLA-27 with a C14-O-(C14) group. Significant immunopharmacological activities were not observed for GLA-58, carrying a C14-O-(C16) group bound to the amino group. GLA-59, carrying 3-O-linked 3-hydroxytetradecanoyl (C14OH) and N-linked C14-O-(C14) groups, showed much higher activities than GLA-27, GLA-60, a compound which possesses the same fatty acid substituents as GLA-59 but with reversed binding sites, showed the strongest B-cell activation and adjuvant activities among the synthetic compounds. Among stereoisomers of GLA-59 and GLA-60 composed of fatty acid substituents with the (RR) and (SS) configuration, compounds with the (RR) configuration elicited stronger activities than the (SS) stereoisomers. The importance of fatty acid substituents, including stereospecificity for the expression of immunopharmacological activities of 4-O

  1. Sphingosine kinase 1 dependent protein kinase C-δ activation plays an important role in acute liver failure in mice

    PubMed Central

    Lei, Yan-Chang; Yang, Ling-Ling; Li, Wen; Luo, Pan

    2015-01-01

    AIM: To investigate the role of protein kinase C (PKC)-δ activation in the pathogenesis of acute liver failure (ALF) in a well-characterized mouse model of D-galactosamine (D-GalN)/lipopolysaccharide (LPS)-induced ALF. METHODS: BALB/c mice were randomly assigned to five groups, and ALF was induced in mice by intraperitoneal injection of D-GaIN (600 mg/kg) and LPS (10 μg/kg). Kaplan-Meier method was used for survival analysis. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels at different time points within one week were determined using a multiparameteric analyzer. Serum levels of high-mobility group box 1 (HMGB1), tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and IL-10 as well as nuclear factor (NF)-κB activity were determined by enzyme-linked immunosorbent assay. Hepatic morphological changes at 36 h after ALF induction were assessed by hematoxylin and eosin staining. Expression of PKC-δ in liver tissue and peripheral blood mononuclear cells (PBMCs) was analyzed by Western blot. RESULTS: The expression and activation of PKC-δ were up-regulated in liver tissue and PBMCs of mice with D-GalN/LPS-induced ALF. Inhibition of PKC-δ activation with rottlerin significantly increased the survival rates and decreased serum ALT/AST levels at 6, 12 and 24 h compared with the control group (P < 0.001). Rottlerin treatment also significantly decreased serum levels of HMGB1 at 6, 12, and 24 h, TNF-α, IL-6 and IL-1 β at 12 h compared with the control group (P < 0.01). The inflammatory cell infiltration and necrosis in liver tissue were also decreased in the rottlerin treatment group. Furthermore, sphingosine kinase 1 (SphK1) dependent PKC-δ activation played an important role in promoting NF-κB activation and inflammatory cytokine production in ALF. CONCLUSION: SphK1 dependent PKC-δ activation plays an important role in promoting NF-κB activation and inflammatory response in ALF, and inhibition of PKC-δ activation might be

  2. Mutational analysis of the redox-sensitive transcriptional regulator OxyR: regions important for oxidation and transcriptional activation.

    PubMed Central

    Kullik, I; Toledano, M B; Tartaglia, L A; Storz, G

    1995-01-01

    OxyR is a redox-sensitive transcriptional regulator of the LysR family which activates the expression of genes important for the defense against hydrogen peroxide in Escherichia coli and Samonella typhimurium. OxyR is sensitive to oxidation and reduction, and only oxidized OxyR is able to activate transcription of its target genes. Using site-directed mutagenesis, we found that one cysteine residue (C-199) is critical for the redox sensitivity of OxyR, and a C-199-->S mutation appears to lock the OxyR protein in the reduced form. We also used a random mutagenesis approach to isolate eight constitutively active mutants. All of the mutations are located in the C-terminal half of the protein, and four of the mutations map near the critical C-199 residue. In vivo as well as in vitro transcription experiments showed that the constitutive mutant proteins were able to activate transcription under both oxidizing and reducing conditions, and DNase I footprints showed that this activation is due to the ability of the mutant proteins to induce cooperative binding of RNA polymerase. Unexpectedly, RNA polymerase was also found to reciprocally affect OxyR binding. PMID:7868602

  3. Comparison of Cr(VI) removal by activated sludge and dissolved organic matter (DOM): importance of UV light.

    PubMed

    Gong, Yu-Feng; Song, Jia; Ren, Hai-Tao; Han, Xu

    2015-12-01

    Removal of toxic Cr(VI) by activated sludge and DOM derived from activated sludge was investigated in this study. A rapid increase in TOC concentration from 50.93 to 127.40 mg L(-1) is observed during the Cr(VI) removal process by activated sludge in the pH range of 2-9. Removal efficiencies of Cr(VI) by either activated sludge or DOM greatly decreased with the increasing initial pH. Kinetics of Cr(VI) removal by activated sludge indicate that both biosorption and bioreduction are involved in the Cr(VI) removal. Cr(VI) removal by DOM is slow in dark, but it is greatly enhanced when UV light is applied. The first-order constant increases from 0.0033 (in dark) to 0.079 min(-1) (UV illumination) at pH 2.0 and 1068 mg L(-1) DOM. The enhancement of Cr(VI) reduction is due to the generation of the reactive intermediates such as O2(●-) and DOM* as DOM absorbed light energy, which plays important roles in the reduction of Cr(VI).

  4. Overexpression of Crocus carotenoid cleavage dioxygenase, CsCCD4b, in Arabidopsis imparts tolerance to dehydration, salt and oxidative stresses by modulating ROS machinery.

    PubMed

    Baba, Shoib Ahmad; Jain, Deepti; Abbas, Nazia; Ashraf, Nasheeman

    2015-09-15

    Apocarotenoids modulate vital physiological and developmental processes in plants. These molecules are formed by the cleavage of carotenoids, a reaction catalyzed by a family of enzymes called carotenoid cleavage dioxygenases (CCDs). Apocarotenoids like β-ionone and β-cyclocitral have been reported to act as stress signal molecules during high light stress in many plant species. In Crocus sativus, these two apocarotenoids are formed by enzymatic cleavage of β-carotene at 9, 10 and 7, 8 bonds by CsCCD4 enzymes. In the present study three isoforms of CsCCD4 were subjected to molecular modeling and docking analysis to determine their substrate specificity and all the three isoforms displayed high substrate specificity for β-carotene. Further, expression of these three CsCCD4 isoforms investigated in response to various stresses revealed that CsCCD4a and CsCCD4b exhibit enhanced expression in response to dehydration, salt and methylviologen, providing a clue towards their role in mediating plant defense response. This was confirmed by overexpressing CsCCD4b in Arabidopsis. The transgenic plants developed longer roots and possessed higher number of lateral roots. Further, overexpression of CsCCD4b imparted enhanced tolerance to salt, dehydration and oxidative stresses as was evidenced by higher survival rate, increased relative root length and biomass in transgenic plants as compared to wild type. Transgenic plants also displayed higher activity and expression of reactive oxygen species (ROS) metabolizing enzymes. This indicates that β-ionone and β-cyclocitral which are enzymatic products of CsCCD4b may act as stress signals and mediate reprogramming of stress responsive genes which ultimately leads to plant defense.

  5. Overexpression of Crocus carotenoid cleavage dioxygenase, CsCCD4b, in Arabidopsis imparts tolerance to dehydration, salt and oxidative stresses by modulating ROS machinery.

    PubMed

    Baba, Shoib Ahmad; Jain, Deepti; Abbas, Nazia; Ashraf, Nasheeman

    2015-09-15

    Apocarotenoids modulate vital physiological and developmental processes in plants. These molecules are formed by the cleavage of carotenoids, a reaction catalyzed by a family of enzymes called carotenoid cleavage dioxygenases (CCDs). Apocarotenoids like β-ionone and β-cyclocitral have been reported to act as stress signal molecules during high light stress in many plant species. In Crocus sativus, these two apocarotenoids are formed by enzymatic cleavage of β-carotene at 9, 10 and 7, 8 bonds by CsCCD4 enzymes. In the present study three isoforms of CsCCD4 were subjected to molecular modeling and docking analysis to determine their substrate specificity and all the three isoforms displayed high substrate specificity for β-carotene. Further, expression of these three CsCCD4 isoforms investigated in response to various stresses revealed that CsCCD4a and CsCCD4b exhibit enhanced expression in response to dehydration, salt and methylviologen, providing a clue towards their role in mediating plant defense response. This was confirmed by overexpressing CsCCD4b in Arabidopsis. The transgenic plants developed longer roots and possessed higher number of lateral roots. Further, overexpression of CsCCD4b imparted enhanced tolerance to salt, dehydration and oxidative stresses as was evidenced by higher survival rate, increased relative root length and biomass in transgenic plants as compared to wild type. Transgenic plants also displayed higher activity and expression of reactive oxygen species (ROS) metabolizing enzymes. This indicates that β-ionone and β-cyclocitral which are enzymatic products of CsCCD4b may act as stress signals and mediate reprogramming of stress responsive genes which ultimately leads to plant defense. PMID:26595090

  6. The importance of delineating networks by activity type in bottlenose dolphins (Tursiops truncatus) in Cedar Key, Florida

    PubMed Central

    Gazda, Stefanie; Iyer, Swami; Killingback, Timothy; Connor, Richard; Brault, Solange

    2015-01-01

    Network analysis has proved to be a valuable tool for studying the behavioural patterns of complex social animals. Often such studies either do not distinguish between different behavioural states of the organisms or simply focus attention on a single behavioural state to the exclusion of all others. In either of these approaches it is impossible to ascertain how the behavioural patterns of individuals depend on the type of activity they are engaged in. Here we report on a network-based analysis of the behavioural associations in a population of bottlenose dolphins (Tursiops truncatus) in Cedar Key, Florida. We consider three distinct behavioural states—socializing, travelling and foraging—and analyse the association networks corresponding to each activity. Moreover, in constructing the different activity networks we do not simply record a spatial association between two individuals as being either present or absent, but rather quantify the degree of any association, thus allowing us to construct weighted networks describing each activity. The results of these weighted activity networks indicate that networks can reveal detailed patterns of bottlenose dolphins at the population level; dolphins socialize in large groups with preferential associations; travel in small groups with preferential associates; and spread out to forage in very small, weakly connected groups. There is some overlap in the socialize and travel networks but little overlap between the forage and other networks. This indicates that the social bonds maintained in other activities are less important as they forage on dispersed, solitary prey. The overall network, not sorted by activity, does not accurately represent any of these patterns. PMID:26064611

  7. Increased aerobic glycolysis is important for the motility of activated VSMC and inhibited by indirubin-3′-monoxime

    PubMed Central

    Heiss, Elke H.; Schachner, Daniel; Donati, Maddalena; Grojer, Christoph S.; Dirsch, Verena M.

    2016-01-01

    Increased aerobic glycolysis is a recognized feature of multiple cellular phenotypes and offers a potential point for drug interference, as pursued by anti-tumor agents targeting the Warburg effect. This study aimed at examining the role of aerobic glycolysis for migration of vascular smooth muscle cells (VSMC) and its susceptibility to the small molecule indirubin-3′-monoxime (I3MO). Activation of VSMC with platelet-derived growth factor (PDGF) resulted in migration and increased glycolytic activity which was accompanied by an increased glucose uptake and hexokinase (HK) 2 expression. Inhibition of glycolysis or hexokinase by pharmacological agents or siRNA-mediated knockdown significantly reduced the migratory behavior in VSMC without affecting cell viability or early actin cytoskeleton rearrangement. I3MO, previously recognized as inhibitor of VSMC migration, was able to counteract the PDGF-activated increase in glycolysis and HK2 abundance. Activation of signal transducer and activator of transcription (STAT) 3 could be identified as crucial event in upregulation of HK2 and glycolytic activity in PDGF-stimulated VSMC and as point of interference for I3MO. I3MO did not inhibit hypoxia-inducible factor (HIF)1α-dependent transcription nor influence miRNA 143 levels, other potential regulators of HK2 levels. Overall, we demonstrate that increased aerobic glycolysis is an important factor for the motility of activated VSMC and that the anti-migratory property of I3MO may partly depend on impairment of glycolysis via a compromised STAT3/HK2 signaling axis. PMID:27185663

  8. The importance of delineating networks by activity type in bottlenose dolphins (Tursiops truncatus) in Cedar Key, Florida.

    PubMed

    Gazda, Stefanie; Iyer, Swami; Killingback, Timothy; Connor, Richard; Brault, Solange

    2015-03-01

    Network analysis has proved to be a valuable tool for studying the behavioural patterns of complex social animals. Often such studies either do not distinguish between different behavioural states of the organisms or simply focus attention on a single behavioural state to the exclusion of all others. In either of these approaches it is impossible to ascertain how the behavioural patterns of individuals depend on the type of activity they are engaged in. Here we report on a network-based analysis of the behavioural associations in a population of bottlenose dolphins (Tursiops truncatus) in Cedar Key, Florida. We consider three distinct behavioural states-socializing, travelling and foraging-and analyse the association networks corresponding to each activity. Moreover, in constructing the different activity networks we do not simply record a spatial association between two individuals as being either present or absent, but rather quantify the degree of any association, thus allowing us to construct weighted networks describing each activity. The results of these weighted activity networks indicate that networks can reveal detailed patterns of bottlenose dolphins at the population level; dolphins socialize in large groups with preferential associations; travel in small groups with preferential associates; and spread out to forage in very small, weakly connected groups. There is some overlap in the socialize and travel networks but little overlap between the forage and other networks. This indicates that the social bonds maintained in other activities are less important as they forage on dispersed, solitary prey. The overall network, not sorted by activity, does not accurately represent any of these patterns.

  9. Pupicidal and repellent activities of Pogostemon cablin essential oil chemical compounds against medically important human vector mosquitoes

    PubMed Central

    Gokulakrishnan, J; Kuppusamy, Elumalai; Shanmugam, Dhanasekaran; Appavu, Anandan; Kaliyamoorthi, Krishnappa

    2013-01-01

    Objective To determine the repellent and pupicidal activities of Pogostemon cablin (P. cablin) chemical compositions were assayed for their toxicity against selected important vector mosquitoes, viz., Aedes aegypti (Ae. aegypti), Anopheles stephensi (An. stephensi) and Culex quinquefasciatus (Cx. quinquefasciatus) (Diptera: Culicidae). Methods The plants dry aerial parts were subjected to hydrodistillation using a modified Clevenger-type apparatus. The composition of the essential oil was analyzed by Gas Chromatography (GC) and GC mass spectrophotometry. Evaluation was carried out in a net cage (45 cm×30 cm×45 cm) containing 100 blood starved female mosquitoes and were assayed in the laboratory condition by using the protocol of WHO 2010. The repellent activity of P. cablin chemical compositions at concentration of 2mg/cm2were applied on skin of fore arm in man and exposed against adult female mosquitoes. The pupicidal activity was determined against selected important vector mosquitoes to concentration of 100 mg/L and mortality of each pupa was recorded after 24 h of exposure to the compounds. Results Chemical constituents of 15 compounds were identified in the oil of P.cablin compounds representing to 98.96%. The major components in essential oil were â-patchoulene, á-guaiene, ã-patchoulene, á-bulnesene and patchouli alcohol. The repellent activity of patchouli alcohol compound was found to be most effective for repellent activity and 2 mg/cm2 concentration provided 100% protection up to 280 min against Ae. aegypti, An. stephensi and Cx. quinquefasciatus, respectively. Similarly, pupae exposed to 100 mg/L concentrations of P. cablin chemical compositions. Among five compounds tested patchouli alcoholwas found to be most effective for pupicidal activity provided 28.44, 26.28 and 25.36 against Ae.aegypti, An.stephensi and Cx. quinquefasciatus, respectively. The percent adult emergence was inversely proportional to the concentration of compounds and directly

  10. Importance of AOX pathway in optimizing photosynthesis under high light stress: role of pyruvate and malate in activating AOX.

    PubMed

    Dinakar, Challabathula; Raghavendra, Agepati S; Padmasree, Kollipara

    2010-05-01

    The present study shows the importance of alternative oxidase (AOX) pathway in optimizing photosynthesis under high light (HL). The responses of photosynthesis and respiration were monitored as O(2) evolution and O(2) uptake in mesophyll protoplasts of pea pre-incubated under different light intensities. Under HL (3000 micromol m(-2) s(-1)), mesophyll protoplasts showed remarkable decrease in the rates of NaHCO(3)-dependent O(2) evolution (indicator of photosynthetic carbon assimilation), while decrease in the rates of respiratory O(2) uptake were marginal. While the capacity of AOX pathway increased significantly by two fold under HL, the capacity of cytochrome oxidase (COX) pathway decreased by >50% compared with capacities under darkness and normal light (NL). Further, the total cellular levels of pyruvate and malate, which are assimilatory products of active photosynthesis and stimulators of AOX activity, were increased remarkably parallel to the increase in AOX protein under HL. Upon restriction of AOX pathway using salicylhydroxamic acid (SHAM), the observed decrease in NaHCO(3)-dependent O(2) evolution or p-benzoquinone (BQ)-dependent O(2) evolution [indicator of photosystem II (PSII) activity] and the increase in total cellular levels of pyruvate and malate were further aggravated/promoted under HL. The significance of raised malate and pyruvate levels in activation of AOX protein/AOX pathway, which in turn play an important role in dissipating excess chloroplastic reducing equivalents and sustenance of photosynthetic carbon assimilation to balance the effects of HL stress on photosynthesis, was depicted as a model.

  11. Antimicrobial Activity of Copaiba (Copaifera officinalis) and Pracaxi (Pentaclethra macroloba) Oils against Staphylococcus Aureus: Importance in Compounding for Wound Care.

    PubMed

    Guimarães, Anna Luísa Aguijar; Cunha, Elisa Alves; Matias, Fernanda Oliveira; Garcia, Patrícia Guedes; Danopoulos, Panagiota; Swikidisa, Rosita; Pinheiro, Vanessa Alves; Nogueira, Rodrigo José Lupatini

    2016-01-01

    The Amazon rainforest is the largest reserve of natural products in the world. Its rich biodiversity of medicinal plants has been utilized by local populations for hundreds of years for the prevention and treatment of various diseases and ailments. Oil extracts from plant species such as Copaifera officinalis and Pentaclethra macroloba are used in compounded formulations for their antiinflammatory, antimicrobial, emollient, moisturizing, and wound-healing activities. The objective of this study was to investigate the in vitro bacteriostatic effect of two Amazonian oils, Copaiba and Pracaxi, against Staphylococcus aureus, a clinically important microorganism responsible for wound infection, to support the use of these oils as novel natural products for compounded wound-treatment modalities. The antibacterial activity of Copaiba and Pracaxi oils against a standard strain of Staphylococcus aureus was assessed using broth microdilution to determine the Minimum Inhibitory Concentration and Minimum Bactericidal Concentration of the oil extracts. Copaiba oil demonstrated antibacterial activity against Staphylococcus aureus, with a Minimum Inhibitory Concentration of 0.3125 mg/mL and a Minimum Bactericidal Concentration of 0.3125 mg/mL. Conversely, Pracaxi oil failed to inhibit Staphylococcus aureus growth. While additional studies are required to further evaluate the antimicrobial activity of Pracaxi oil, even low concentrations of Copaiba oil effectively inhibited Staphylococcus aureus growth, supporting its potential use as a promising adjuvant in compounded topical formulations for wound and scar healing. PMID:27125055

  12. Importance of sulfate groups for the macrophage-stimulating activities of ascophyllan isolated from the brown alga Ascophyllum nodosum.

    PubMed

    Jiang, Zedong; Ueno, Mikinori; Nishiguchi, Tomoki; Abu, Ryogo; Isaka, Shogo; Okimura, Takasi; Yamaguchi, Kenichi; Oda, Tatsuya

    2013-10-18

    To investigate the role of sulfate groups on the macrophage-stimulating activities of ascophyllan, we prepared desulfated ascophyllan, and its effects on RAW264.7 cells were compared with native ascophyllan. The chemical structural analysis revealed that nearly 21% of sulfate groups of ascophyllan were removed by desulfation reaction, while no significant changes in the molecular mass and monosaccharide composition occurred after desulfation. NO- and cytokine- (TNF-α and G-CSF) inducing activities of the desulfated ascophyllan on RAW264.7 cells were significantly decreased as compared to native ascophyllan. Furthermore, the activity of desulfated ascophyllan to induce reactive oxygen species (ROS) generation from RAW264.7 cells decreased to almost negligible level. Our results suggest that the level of sulfate groups of ascophyllan is an important structural element responsible for the macrophage-stimulating activities. Probably, even the limited removal of sulfate residues sensitive to desulfation reaction may result in significant decrease in the bioactivities of ascophyllan. PMID:24025707

  13. Notch and Wnt/β-catenin signaling pathway play important roles in activating liver cancer stem cells

    PubMed Central

    Wang, Ronghua; Sun, Qian; Wang, Peng; Liu, Man; Xiong, Si; Luo, Jing; Huang, Hai; Du, Qiang; Geller, David A.; Cheng, Bin

    2016-01-01

    Human hepatocellular carcinoma (HCC) is driven and maintained by liver cancer stem cells (LCSCs) that display stem cell properties. These LCSCs are promoted by the intersecting of Notch and Wnt/β-Catenin signaling pathways. In this study, we demonstrate that LCSCs with markers CD90, CD24, CD13, and CD133 possess stem properties of self-renewal and tumorigenicity in NOD/SCID mice. The increased expression of these markers was correlated with advanced disease stage, larger tumors, and worse overall survival in 61 HCC cases. We also found that both Notch and Wnt/β-catenin signaling pathways played important roles in increasing the stem-ness characteristics of LCSCs. Our data suggested that Notch1 was downstream of Wnt/β-catenin. The active form of Notch1 intracellular domain (NICD) expression depended on Wnt/β-catenin pathway activation. Moreover, Notch1 negatively contributed to Wnt/β-catenin signaling modulation. Knock down of Notch1 with lentivirus N1ShRNA up-regulated the active form of β-catenin. Ectopic expression of NICD with LV-Notch1 in LCSCs attenuated β-catenin/TCF dependent luciferase activity significantly. In addition, there was a non-proteasome mediated feedback loop between Notch1 and Wnt/β-catenin signaling in LCSCs. The central role of Notch and the Wnt/β-catenin signaling pathway in LCSCs may provide an attractive therapeutic strategy against HCC. PMID:26735577

  14. Neurotoxicity and other pharmacological activities of the snake venom phospholipase A2 OS2: The N-terminal region is more important than enzymatic activity

    PubMed Central

    Rouault, Morgane; Rash, Lachlan D.; Escoubas, Pierre; Boilard, Eric; Bollinger, James; Lomonte, Bruno; Maurin, Thomas; Guillaume, Carole; Canaan, Stéphane; Deregnaucourt, Christiane; Schrével, Joseph; Doglio, Alain; Gutiérrez, José María; Lazdunski, Michel; Gelb, Michael H.; Lambeau, Gérard

    2009-01-01

    Several snake venom secreted phospholipases A2 (sPLA2s) including OS2 exert a variety of pharmacological effects ranging from central neurotoxicity to anti-HIV activity by mechanisms that are not yet fully understood. To conclusively address the role of enzymatic activity and map the key structural elements of OS2 responsible for its pharmacological properties, we have prepared single point OS2 mutants at the catalytic site and large chimeras between OS2 and OS1, an homologous but non toxic sPLA2. Most importantly, we found that the enzymatic activity of the active site mutant H48Q is 500-fold lower than that of the wild-type protein, while central neurotoxicity is only 16-fold lower, providing convincing evidence that catalytic activity is at most a minor factor that determines central neurotoxicity. The chimera approach has identified the N-terminal region (residues 1–22) of OS2, but not the central one (residues 58–89), as crucial for both enzymatic activity and pharmacological effects. The C-terminal region of OS2 (residues 102–119) was found to be critical for enzymatic activity, but not for central neurotoxicity and anti-HIV activity, allowing us to further dissociate enzymatic activity and pharmacological effects. Finally, direct binding studies with the C-terminal chimera which poorly binds to phospholipids while it is still neurotoxic, led to the identification of a subset of brain N-type receptors which may be directly involved in central neurotoxicity. PMID:16669624

  15. Nucleoporin 62-Like Protein Activates Canonical Wnt Signaling through Facilitating the Nuclear Import of β-Catenin in Zebrafish

    PubMed Central

    Yang, Xiaojie; Gu, Qilin; Lin, Li; Li, Shaoyang; Zhong, Shan

    2015-01-01

    Nucleoporin p62 (Nup62) localizes in the central channel of nuclear pore complexes (NPCs) and regulates nuclear pore permeability and nucleocytoplasmic transport. However, the developmental roles of Nup62 in vertebrates remain largely unclear. Zebrafish Nup62-like protein (Nup62l) is a homolog of mammalian Nup62. The nup62l gene is maternally expressed, but its transcripts are ubiquitously distributed during early embryogenesis and enriched in the head, pharynx, and intestine of developing embryos. Activation of the Wnt/β-catenin pathway positively modulates nup62l transcription, while Bmp signaling acts downstream of Wnt/β-catenin signaling to negatively regulate nup62l expression. Overexpression of nup62l dorsalized embryos and enhanced gastrula convergence and extension (CE) movements. In contrast, knockdown of Nup62l led to ventralized embryos, an impediment to CE movements, and defects in specification of midline organ progenitors. Mechanistically, Nup62l acts as an activator of Wnt/β-catenin signaling through interaction with and facilitation of nuclear import of β-catenin-1/2 in zebrafish. Thus, Nup62l regulates dorsoventral patterning, gastrula CE movements, and proper specification of midline organ precursors through mediating the nuclear import of β-catenins in zebrafish. PMID:25605329

  16. The Importance of Stereochemically Active Lone Pairs For Influencing Pb(II) and As(III) Protein Binding

    PubMed Central

    Neupane, Kosh P.; De Gioia, Luca

    2012-01-01

    Heavy metal toxicity is a worldwide problem which is associated with the metal’s high affinity for thiolate rich proteins. Despite the tremendous toxicity concern, the mode of binding of As(III) and Pb(II) to proteins is poorly understood. To clarify the requirements for toxic metal binding to metalloregulatory sensor proteins such as As(III) in ArsR/ArsD and Pb(II) in PbrR or replacing Zn(II) in δ-aminolevulinc acid dehydratase (ALAD), we have employed computational and experimental methods examining these heavy metals binding to designed peptide models. The computational results show that the mode of coordination of As(III) and Pb(II) is greatly influenced by the steric bulk within the second coordination environment of the metal. The proposed basis of this selectivity is the large size of the ion and, most important, the influence of the stereochemically active lone pair in hemi-directed complexes of the metal ion as being crucial. The experimental data show that switching a bulky leucine layer above the metal binding site by a smaller alanine residue enhances the Pb(II) binding affinity by a factor of five supporting experimentally this lone pair steric hindrance hypothesis. These complementary approaches demonstrate the potential importance of a stereochemically active lone pair as a metal recognition mode in proteins and, specifically, how the second coordination sphere environment affects the affinity and selectivity of protein targets by certain toxic ions. PMID:22231489

  17. Importance of the tuning of band position in optimizing the electronic coupling and photocatalytic activity of nanocomposite

    SciTech Connect

    Jin, Xiaoyan; Mok, Eun Kyung; Baek, Ji-Won; Park, Sang-Hyun; Hwang, Seong-Ju

    2015-10-15

    The electronic coupling and photocatalytic activity of Ag{sub 2}CO{sub 3}–TiO{sub 2} nanocomposite can be optimized by the fine-tuning of the band position of titanium oxide with nitrogen doping. The increase of the valence band energy of TiO{sub 2} by N-doping leads not only to the enhanced absorption of visible light but also to the promoted hole transfer from Ag{sub 2}CO{sub 3} to TiO{sub 2}, resulting in the efficient spatial separation of photogenerated electrons and holes. While the undoped Ag{sub 2}CO{sub 3}–TiO{sub 2} nanocomposite shows an inferior photocatalytic activity to the pure Ag{sub 2}CO{sub 3}, the photocatalyst performance of N-doped nanocomposite is better than those of Ag{sub 2}CO{sub 3} and undoped Ag{sub 2}CO{sub 3}–TiO{sub 2} nanocomposite. This observation underscores a significant enhancement of the photocatalytic activity of nanocomposite upon N-doping, a result of enhanced electronic coupling between the hybridized species. The present results clearly demonstrate the importance of the fine-tuning of band position in optimizing the photocatalytic activity of hybrid-type photocatalysts. - Highlights: • The band position of Ag{sub 2}CO{sub 3}–TiO{sub 2} can be effectively tailored by nitrogen doping. • The N-doping leads to the improvement of charge separation. • The N-doped Ag{sub 2}CO{sub 3}–TiO{sub 2} shows high photocatalytic activity.

  18. Fear of Movement and Low Self-Efficacy Are Important Barriers in Physical Activity after Renal Transplantation

    PubMed Central

    Zelle, Dorien M.; Corpeleijn, Eva; Klaassen, Gerald; Schutte, Elise; Navis, Gerjan; Bakker, Stephan J. L.

    2016-01-01

    Background Physical activity (PA) and exercise are commonly used as preventive measures for cardiovascular disease in the general population, and could be effective in the management of post-transplantation cardiovascular risk. PA levels are low after renal transplantation and very few renal transplant recipients (RTR) meet the PA guidelines. Identification of barriers to regular PA is important to identify targets for intervention to improve PA levels after renal transplantation. We investigated fear of movement and physical self-efficacy as barriers to PA in RTR. Methods RTR were investigated between 2001–2003. The Tampa Score of Kinesiophobia–Dutch Version (TSK-11) was used to assess fear of movement. Physical self-efficacy was measured with the LIVAS-scale. PA was assessed using validated questionnaires (Tecumseh Occupational Activity Questionnaire and the Minnesota Leisure Time Physical Activity Questionnaire). Results A total of 487 RTR (age 51±12 years, 55% men) were studied. Median score [interquartile range] on TSK-11 was 22 [17–26]. Low physical self-efficacy (Exp B:0.41[0.31–0.54], p<0.001) and history of myocardial infarction, transient ischemic attack and cerebrovascular accident (Exp B:1.30[1.03–1.63],p = 0.03) were independent determinants for fear of movement. Fear of movement was associated with lower daily PA, occupational, sports and leisure time PA. Mediation-analysis showed that a large part (73%) of the effect of fear of movement on PA was explained by low physical self-efficacy. Conclusions This study was the first to examine fear of movement and self-efficacy in relation to PA in RTR. Fear of movement was associated with a low PA level, and the larger part of this relation was mediated by low physical self-efficacy. Both fear of movement and physical self-efficacy level are important targets for intervention during rehabilitation after renal transplantation. PMID:26844883

  19. Towards Pro-active Embodied Agents: On the Importance of Neural Mechanisms Suitable to Process Time Information

    NASA Astrophysics Data System (ADS)

    de Croon, G.; Nolfi, S.; Postma, E. O.

    In Embodied Cognitive Science, many studies have focused on reactive agents, i.e. agents that have no internal state and always respond in the same way to the same stimulus. However, this particular focus is not due to a rejection of the importance of internal states. Rather, it is due to the difficulty of developing pro-active embodied and situated agents, that is agents able to: (a) extract internal states by integrating sensorymotor information through time and, (b) later use these internal states to modulate their motor behaviour according to the current environmental circumstances. In this chapter we will focus on how pro-active agents can be developed and, more specifically, on which are the neural mechanisms that might favour the development of pro-active agents. By comparing the results of five sets of evolutionary experiments in which simulated robots are provided with different types of recurrent neural networks, we gain insight into the relation between the robots` capabilities and the characteristics of their neural controllers. We show how special mechanisms for processing information in time facilitate the exploitation of internal states.

  20. Hypothalamic-pituitary-adrenocortical (HPA) activity in kindergarten children: importance of gender and associations with behavioral/emotional difficulties.

    PubMed

    Hatzinger, Martin; Brand, Serge; Perren, Sonja; von Wyl, Agnes; von Klitzing, Kai; Holsboer-Trachsler, Edith

    2007-11-01

    The current cross-sectional study investigated basal and stress-challenged hypothalamic-pituitary-adrenocortical (HPA) system function in 102 five-year old kindergarten children (59 boys, 43 girls) who had been assessed by a comprehensive psychological and behavioral test battery. Baseline HPA system activity was significantly increased in girls when compared to boys (p<0.001). Furthermore, basal HPA system activity predicted a high hormonal release during stress with--again--girls showing higher hormonal responses than boys (p<0.01). Importantly, increased HPA system activity (baseline and stress-challenged) was significantly associated with hyperactivity/impulsivity and emotional problems in boys and with positive emotions in girls (p<0.05). These results suggest an occurrence of neurobiological alterations early in development. The observed neurobiological changes are gender specific already at the age of 5 years. Prospective long-term follow up of the identified subjects with HPA axis alterations will clarify if these markers are predictive for the onset of psychiatric disorders. PMID:16979188

  1. Type I beta-turn conformation is important for biological activity of the melanocyte-stimulating hormone analogues.

    PubMed

    Li, S Z; Lee, J H; Lee, W; Yoon, C J; Baik, J H; Lim, S K

    1999-10-01

    In order to define which structure of alpha-melanocyte-stimulating hormone (MSH) analogues plays a critical role for ligand-receptor interaction and selectivity, we analysed receptor-binding and cAMP-generating activity in Chinese hamster ovary cell lines stably transfected with rMC3R and hMC4R, as well as the NMR structures of chemically synthesized alpha-MSH analogues. Compared with [Ahx4]alpha-MSH, the linear MTII designated as alpha-MSH-ND revealed a preference for the MC4R, whereas its IC50 and EC50 values were comparable to those of MTII reported previously. Truncation of Ahx4 and Asp5 of alpha-MSH-ND remarkably decreased the receptor-binding and cAMP-generating activity. Meanwhile, maximum cAMP-generating activity was observed at a higher concentration (10(-5) M) of alpha-MSH-ND(6-10), and MC4R preference was changed into MC3R preference. In contrast, [Gln6]alpha-MSH-ND(6-10) lost its cAMP-generating activity almost completely, even though it bound to both receptors. Whereas the solution conformation of alpha-MSH-ND revealed a stable type I beta-turn structure, [Gln6]alpha-MSH-ND(6-10) revealed a tight gamma-turn composed of Gln6-D-Phe7-Arg8. Replacement of the His6 residue of alpha-MSH-ND by Gln, Asn, Arg or Lys decreased not only the receptor binding, but also the cAMP-generating activity in both the MC3R and the MC4R. The structure of [Gln6]alpha-MSH-ND exhibited a stable type I' beta-turn comprising Asp5, Gln6, D-Phe7 and Arg8. [Lys6]alpha-MSH-ND showed a greatly reduced binding affinity and cAMP-generating activity with the loss of MC4R selectivity. In NMR studies, [Lys6]alpha-MSH-ND also demonstrated a gamma-turn conformation around Lys6-DPhe7-Arg8. From the above results, we conclude that a type I beta-turn conformation comprising the residues Asp5-His6-(D-Phe7)-Arg8 was important for receptor binding and activation, as well as the selectivity of MSH analogues.

  2. Autophagy is not required to sustain exercise and PRKAA1/AMPK activity but is important to prevent mitochondrial damage during physical activity.

    PubMed

    Lo Verso, Francesca; Carnio, Silvia; Vainshtein, Anna; Sandri, Marco

    2014-01-01

    Physical activity has been recently documented to play a fundamental physiological role in the regulation of autophagy in several tissues. It has also been reported that autophagy is required for exercise itself and for training-induced adaptations in glucose homeostasis. These autophagy-mediated metabolic improvements are thought to be largely dependent on the activation of the metabolic sensor PRKAA1/AMPK. However, it is unknown whether these important benefits stem from systemic adaptations or are due solely to alterations in skeletal muscle metabolism. To address this we utilized inducible, muscle-specific, atg7 knockout mice that we have recently generated. Our findings indicate that acute inhibition of autophagy in skeletal muscle just prior to exercise does not have an impact on physical performance, PRKAA1 activation, or glucose homeostasis. However, we reveal that autophagy is critical for the preservation of mitochondrial function during damaging muscle contraction. This effect appears to be gender specific affecting primarily females. We also establish that basal oxidative stress plays a crucial role in mitochondrial maintenance during normal physical activity. Therefore, autophagy is an adaptive response to exercise that ensures effective mitochondrial quality control during damaging physical activity. PMID:25483961

  3. Gingerols and shogaols: Important nutraceutical principles from ginger.

    PubMed

    Semwal, Ruchi Badoni; Semwal, Deepak Kumar; Combrinck, Sandra; Viljoen, Alvaro M

    2015-09-01

    Gingerols are the major pungent compounds present in the rhizomes of ginger (Zingiber officinale Roscoe) and are renowned for their contribution to human health and nutrition. Medicinal properties of ginger, including the alleviation of nausea, arthritis and pain, have been associated with the gingerols. Gingerol analogues are thermally labile and easily undergo dehydration reactions to form the corresponding shogaols, which impart the characteristic pungent taste to dried ginger. Both gingerols and shogaols exhibit a host of biological activities, ranging from anticancer, anti-oxidant, antimicrobial, anti-inflammatory and anti-allergic to various central nervous system activities. Shogaols are important biomarkers used for the quality control of many ginger-containing products, due to their diverse biological activities. In this review, a large body of available knowledge on the biosynthesis, chemical synthesis and pharmacological activities, as well as on the structure-activity relationships of various gingerols and shogaols, have been collated, coherently summarised and discussed. The manuscript highlights convincing evidence indicating that these phenolic compounds could serve as important lead molecules for the development of therapeutic agents to treat various life-threatening human diseases, particularly cancer. Inclusion of ginger or ginger extracts in nutraceutical formulations could provide valuable protection against diabetes, cardiac and hepatic disorders. PMID:26228533

  4. Gingerols and shogaols: Important nutraceutical principles from ginger.

    PubMed

    Semwal, Ruchi Badoni; Semwal, Deepak Kumar; Combrinck, Sandra; Viljoen, Alvaro M

    2015-09-01

    Gingerols are the major pungent compounds present in the rhizomes of ginger (Zingiber officinale Roscoe) and are renowned for their contribution to human health and nutrition. Medicinal properties of ginger, including the alleviation of nausea, arthritis and pain, have been associated with the gingerols. Gingerol analogues are thermally labile and easily undergo dehydration reactions to form the corresponding shogaols, which impart the characteristic pungent taste to dried ginger. Both gingerols and shogaols exhibit a host of biological activities, ranging from anticancer, anti-oxidant, antimicrobial, anti-inflammatory and anti-allergic to various central nervous system activities. Shogaols are important biomarkers used for the quality control of many ginger-containing products, due to their diverse biological activities. In this review, a large body of available knowledge on the biosynthesis, chemical synthesis and pharmacological activities, as well as on the structure-activity relationships of various gingerols and shogaols, have been collated, coherently summarised and discussed. The manuscript highlights convincing evidence indicating that these phenolic compounds could serve as important lead molecules for the development of therapeutic agents to treat various life-threatening human diseases, particularly cancer. Inclusion of ginger or ginger extracts in nutraceutical formulations could provide valuable protection against diabetes, cardiac and hepatic disorders.

  5. Sterol-Dependent Nuclear Import of ORP1S Promotes LXR Regulated Trans-Activation of APOE

    PubMed Central

    Lee, Sungsoo; Wang, Ping-Yuan; Jeong, Yangsik; Mangelsdorf, David J.; Anderson, Richard G. W.; Michaely, Peter

    2013-01-01

    Oxysterol binding protein related protein 1S (ORP1S) is a member of a family of sterol transport proteins. Here we present evidence that ORP1S translocates from the cytoplasm to the nucleus in response to sterol binding. The sterols that best promote nuclear import of ORP1S also activate the liver X receptor (LXR) transcription factors and we show that ORP1S binds to LXRs, promotes binding of LXRs to LXR response elements (LXREs) and specifically enhances LXR-dependent transcription via the ME.1 and ME.2 enhancer elements of the apoE gene. We propose that ORP1S is a cytoplasmic sterol sensor, which transports sterols to the nucleus and promotes LXR-dependent gene transcription through select enhancer elements. PMID:22728266

  6. Terracidiphilus gabretensis gen. nov., sp. nov., an Abundant and Active Forest Soil Acidobacterium Important in Organic Matter Transformation

    PubMed Central

    García-Fraile, Paula; Benada, Oldrich; Cajthaml, Tomáš; Baldrian, Petr

    2015-01-01

    Understanding the activity of bacteria in coniferous forests is highly important, due to the role of these environments as a global carbon sink. In a study of the microbial biodiversity of montane coniferous forest soil in the Bohemian Forest National Park (Czech Republic), we succeeded in isolating bacterial strain S55T, which belongs to one of the most abundant operational taxonomic units (OTUs) in active bacterial populations, according to the analysis of RNA-derived 16S rRNA amplicons. The 16S rRNA gene sequence analysis showed that the species most closely related to strain S55T include Bryocella elongata SN10T (95.4% identity), Acidicapsa ligni WH120T (95.2% identity), and Telmatobacter bradus TPB6017T (95.0% identity), revealing that strain S55T should be classified within the phylum Acidobacteria, subdivision 1. Strain S55T is a rod-like bacterium that grows at acidic pH (3 to 6). Its phylogenetic, genotypic, phenotypic, and chemotaxonomic characteristics indicate that strain S55T corresponds to a new genus within the phylum Acidobacteria; thus, we propose the name Terracidiphilus gabretensis gen. nov., sp. nov. (strain S55T = NBRC 111238T = CECT 8791T). This strain produces extracellular enzymes implicated in the degradation of plant-derived biopolymers. Moreover, analysis of the genome sequence of strain S55T also reveals the presence of enzymatic machinery required for organic matter decomposition. Soil metatranscriptomic analyses found 132 genes from strain S55T being expressed in the forest soil, especially during winter. Our results suggest an important contribution of T. gabretensis S55T in the carbon cycle in the Picea abies coniferous forest. PMID:26546425

  7. [Importance of the hyperuricaemia, gout and gender nosological features in the activity of general practitioner - family doctor].

    PubMed

    Rudichenko, V M

    2012-01-01

    In this article there were analyzed gender data about features of hyperuricaemia and gout: women are much older at the onset of gout arthritis (one of main reasons, probably, makes menopause by itself), have more associated comorbid deseases as hypertension and kidney failure and drinks less alcoholic beverages. It was noticed, that typical localisation of the lesion on the first toe is less often in women, and women are more inclined to use diuretics among medical drugs. Abovementioned clinical features are of some importance for the broad activity of general practitioners - family doctors. Gender features of polyarthicular gout are not uniformed. Scientific researches confirmed possibility of the genetic basis of the uric acid metabolism, which influences some fenotypical features of the organism. Several genes are known for their influence on serum uric acid: PDZK1, GCKR, SLC2A9, ABCG2, LRRC16A, SLC17A3, SLC16A9 and SLC22A12. However, conclusions of the research works confirm the necessity of scientific clarification of the importance of different factors of gender differences.

  8. Sterol-dependent nuclear import of ORP1S promotes LXR regulated trans-activation of apoE

    SciTech Connect

    Lee, Sungsoo; Wang, Ping-Yuan; Jeong, Yangsik; Mangelsdorf, David J.; Anderson, Richard G.W.; Michaely, Peter

    2012-10-01

    Oxysterol binding protein related protein 1S (ORP1S) is a member of a family of sterol transport proteins. Here we present evidence that ORP1S translocates from the cytoplasm to the nucleus in response to sterol binding. The sterols that best promote nuclear import of ORP1S also activate the liver X receptor (LXR) transcription factors and we show that ORP1S binds to LXRs, promotes binding of LXRs to LXR response elements (LXREs) and specifically enhances LXR-dependent transcription via the ME.1 and ME.2 enhancer elements of the apoE gene. We propose that ORP1S is a cytoplasmic sterol sensor, which transports sterols to the nucleus and promotes LXR-dependent gene transcription through select enhancer elements. -- Highlights: Black-Right-Pointing-Pointer ORP1S translocates to the nucleus in response to sterol binding. Black-Right-Pointing-Pointer The sterols that best promote nuclear import of ORP1S are LXR agonists. Black-Right-Pointing-Pointer ORP1S binds to LXRs, enhances binding of LXRs to LXREs and promotes LXR-dependent transcription of apoE.

  9. TGF-β: An Important Mediator of Allergic Disease and a Molecule with Dual Activity in Cancer Development

    PubMed Central

    Tirado-Rodriguez, Belen; Segura-Medina, Patricia; Huerta-Yepez, Sara

    2014-01-01

    The transforming growth factor-β (TGF-β) superfamily is a family of structurally related proteins that includes TGF-β, activins/inhibins, and bone morphogenic proteins (BMPs). Members of the TGF-β superfamily regulate cellular functions such as proliferation, apoptosis, differentiation, and migration and thus play key roles in organismal development. TGF-β is involved in several human diseases, including autoimmune disorders and vascular diseases. Activation of the TGF-β receptor induces phosphorylation of serine/threonine residues and triggers phosphorylation of intracellular effectors (Smads). Once activated, Smad proteins translocate to the nucleus and induce transcription of their target genes, regulating various processes and cellular functions. Recently, there has been an attempt to correlate the effect of TGF-β with various pathological entities such as allergic diseases and cancer, yielding a new area of research known as “allergooncology," which investigates the mechanisms by which allergic diseases may influence the progression of certain cancers. This knowledge could generate new therapeutic strategies aimed at correcting the pathologies in which TGF-β is involved. Here, we review recent studies that suggest an important role for TGF-β in both allergic disease and cancer progression. PMID:25110717

  10. Arachnids of medical importance in Brazil: main active compounds present in scorpion and spider venoms and tick saliva.

    PubMed

    Cordeiro, Francielle A; Amorim, Fernanda G; Anjolette, Fernando A P; Arantes, Eliane C

    2015-01-01

    Arachnida is the largest class among the arthropods, constituting over 60,000 described species (spiders, mites, ticks, scorpions, palpigrades, pseudoscorpions, solpugids and harvestmen). Many accidents are caused by arachnids, especially spiders and scorpions, while some diseases can be transmitted by mites and ticks. These animals are widely dispersed in urban centers due to the large availability of shelter and food, increasing the incidence of accidents. Several protein and non-protein compounds present in the venom and saliva of these animals are responsible for symptoms observed in envenoming, exhibiting neurotoxic, dermonecrotic and hemorrhagic activities. The phylogenomic analysis from the complementary DNA of single-copy nuclear protein-coding genes shows that these animals share some common protein families known as neurotoxins, defensins, hyaluronidase, antimicrobial peptides, phospholipases and proteinases. This indicates that the venoms from these animals may present components with functional and structural similarities. Therefore, we described in this review the main components present in spider and scorpion venom as well as in tick saliva, since they have similar components. These three arachnids are responsible for many accidents of medical relevance in Brazil. Additionally, this study shows potential biotechnological applications of some components with important biological activities, which may motivate the conducting of further research studies on their action mechanisms.

  11. Membrane permeability of redox active metal chelators: an important element in reducing hydroxyl radical induced NAD+ depletion in neuronal cells.

    PubMed

    Jayasena, T; Grant, R S; Keerthisinghe, N; Solaja, I; Smythe, G A

    2007-03-01

    There is substantial evidence implicating increased production of the hydroxyl radical and oxidative stress in the pathogenesis of neurodegenerative diseases such as Alzheimer's disease (AD). Significant amounts of hydroxyl radicals will be produced in the presence of hydrogen peroxide and redox active iron via Fenton chemistry. Increased iron levels within the cytoplasm of vulnerable neurons suggest that this may also be an important site of oxidative activity. We investigated the likelihood that intracellular, rather than extracellular chelation of ferrous or ferric iron may be more effective in reducing hydroxyl radical induced cell damage and preserving NAD(+) levels and cell viability. Using intracellular NAD(H) measurements as an indicator of cell viability we found that membrane permeable ferrous chelators were most efficient in preserving cellular NAD(+) levels. Hydrophilic, ferrous or ferric chelators and lipophilic ferric chelators were essentially ineffective in preventing cellular NAD(+) depletion when added at physiological concentrations. We propose that lipophilic ferrous chelators, due to their actions inside the cell, are effective agents for moderating neuronal damage in conditions such as AD where intracellular oxidative stress plays a significant role in disease pathology. PMID:17210195

  12. Terpenoid composition and antifungal activity of three commercially important essential oils against Aspergillus flavus and Aspergillus niger.

    PubMed

    Bisht, Deepa; Pal, Anirban; Chanotiya, C S; Mishra, Dhirendra; Pandey, K N

    2011-12-01

    Hydro-distilled essential oils extracted from three commercially important aromatic plants were analysed by capillary gas chromatography-flame ionization detector and gas chromatography/quadrupole mass spectrometry and subjected to antifungal activity. Fifteen compounds, which accounted for 97.8% of Acorus calamus root oil composition have been identified. Besides the major constituent (Z)-asarone (81.1-92.4%), (Z)-methyl isoeugenol (1.8-2.1%), (Z)-isoelemicin (1.2-1.3%), (E)-asarone (1.0-2.6%), (E)-methyl isoeugenol (0.2-0.4%), (Z)-β-ocimene (0.2-0.4%), elemicin (0.2-0.3%), linalool (0.1-0.9%) and kessane (t-0.2%) were identified. Monoterpenes constituted the main fraction of Origanum vulgare essential oil attaining 90.5% of the total oil composition. p-Cymene (10.3%) was the major component of the monoterpene hydrocarbon fraction while thymol (53.2%) and carvacrol (3.9%) were the most abundant oxygenated monoterpenes among the 33 identified constituents. Cinnamomum tamala leaf oil contained (E)-cinnamaldehyde as the principal component. Quantitative variations in (Z)-cinnamaldehyde (5.8-7.1%), linalool (6.4-8.5%) and (E)-cinnamyl acetate (4.7-5.2%) were significant. The antifungal activity of the hydro-distilled essential oils of A. calamus, O. vulgare and C. tamala were evaluated against Aspergillus flavus and Aspergillus niger. Disc diffusion method was used for the determination of the inhibitory effect. O. vulgare essential oil exhibited the highest activity. Moreover, all three essential oils inhibit the growth of A. flavus and A. niger. PMID:21707253

  13. Effect of Erica sp. honey against microorganisms of clinical importance: study of the factors underlying this biological activity.

    PubMed

    Feás, Xesus; Iglesias, Antonio; Rodrigues, Sandra; Estevinho, Leticia M

    2013-04-11

    This study aimed to determine the factors (phenolic compounds, flavonoids, sugars or H2O2) that contribute the most to the antimicrobial activity of heather honey samples against four yeasts and four bacteria with medical importance. To discard the effect of H2O2 in the antimicrobial activity, catalase was added. To evaluate the osmotic pressure's effect, artificial honey was also used. Phenolic compounds and flavonoids were determined and Pearson's correlation analysis was performed to assess whether these correlated with antimicrobial activity. The amount of phenolic compounds ranged from 630.89 ± 5.21 GAE kg-1 to 718.92 ± 4.41 GAE kg-1, while the flavonoids varied between 450.72 ± 5.67 CAE kg-1 and 673.98 ± 4.33 CAE kg-1. For the bacteria, the minimum inhibitory concentration (MIC) of the honey without catalase ranged from 1.01 ± 0.50% to 10.00 ± 4.72% and was between 2.00 ± 0.94% and 13.27 ± 5.23% for honey with catalase. Concerning the yeasts, the MICs was between 13.16 ± 4.08% and 20.00 ± 5.09% for honey without catalase and between 14.95 ± 4.16% and 25.67 ± 5.50% for honey with catalase. The elucidation of the antimicrobial factors and action mechanisms is essential for the correct use of honey in therapeutic applications.

  14. Cooperative effects of hepatitis B virus and TNF may play important roles in the activation of metabolic pathways through the activation of NF-κB

    PubMed Central

    Wu, Shuang; Kanda, Tatsuo; Nakamoto, Shingo; Jiang, Xia; Nakamura, Masato; Sasaki, Reina; Haga, Yuki; Shirasawa, Hiroshi; Yokosuka, Osamu

    2016-01-01

    Elevated levels of inflammatory cytokines such as tumor necrosis factor-α (TNF-α) and interleukin (IL)-1β are often observed in the sera of hepatitis B virus (HBV)-infected patients. It is well known that these cytokines activate nuclear factor-κB (NF-κB)-signaling, and are associated with endoplasmic reticulum (ER) stress. We investigated whether HBV or HBV X protein (HBx) enhanced the activation of NF-κB in the presence of TNF and/or IL-1β, and their effects on the expression of metabolic pathway-associated genes. We examined whether HBV or HBx enhanced cytokine-induced activation of NF-κB in hepatocytes, using a reporter assay, in the presence or absence of TNF and/or IL-1β. The expression of insulin-like growth factor binding protein 1 (IGFBP1), one of the NF-κB target genes was also examined. The expression of metabolic pathway-associated genes in HepG2 and HepG2.2.15 cells in the presence or absence of TNF was evaluated by RT-qPCR. Human hepatocytes expressed TNF receptors and IL-1 receptors. NF-κB was activated by cooperation between HBx and TNF in human hepatocytes. We observed IGFBP1 expression in HBV infection and that a number of metabolic pathway-associated genes were upregulated in HepG2.2.15 cells, compared with HepG2 cells with or without TNF treatment. We observed the cooperative effects of HBV and TNF which enhanced the activation of NF-κB as well as upregulated the expression of metabolic pathway-associated genes in hepatocytes. These effects may be important in the development of HBV-associated metabolic syndrome. PMID:27315566

  15. Effective ATPase activity and moderate chaperonin-cochaperonin interaction are important for the functional single-ring chaperonin system.

    PubMed

    Illingworth, Melissa; Salisbury, Jared; Li, Wenqian; Lin, Donghai; Chen, Lingling

    2015-10-01

    Escherichia coli chaperonin GroEL and its cochaperonin GroES are essential for cell growth as they assist folding of cellular proteins. The double-ring assembly of GroEL is required for the chaperone function, and a single-ring variant GroEL(SR) is inactive with GroES. Mutations in GroEL(SR) (A92T, D115N, E191G, and A399T) have been shown to render GroEL(SR)-GroES functional, but the molecular mechanism of activation is unclear. Here we examined various biochemical properties of these functional GroEL(SR)-GroES variants, including ATP hydrolysis rate, chaperonin-cochaperonin interaction, and in vitro protein folding activity. We found that, unlike the diminished ATPase activity of the inactive GroEL(SR)-GroES, all four single-ring variants hydrolyzed ATP at a level comparable to that of the double-ring GroEL-GroES. The chaperonin-cochaperonin interaction in these single-ring systems was weaker, by at least a 50-fold reduction, than the highly stable inactive GroEL(SR)-GroES. Strikingly, only GroEL(SR)D115N-GroES and GroEL(SR)A399T-GroES assisted folding of malate dehydrogenase (MDH), a commonly used folding substrate. These in vitro results are interesting considering that all four of the single-ring systems were able to substitute GroEL-GroES to support cell growth, suggesting that the precise action of chaperonin on MDH folding may not represent that on the intrinsic cellular substrates. Our findings that both effective ATP hydrolysis rate and moderate chaperonin-cochaperonin interaction are important factors for functional single-ring GroEL(SR)-GroES are reminiscent of the naturally occurring single-ring human mitochondrial chaperonin mtHsp60-mtHsp10. Differences in biochemical properties between the single- and double-ring chaperonin systems may be exploited in designing molecules for selective targeting. PMID:26271593

  16. Alumoxane/ferroxane nanoparticles for the removal of viral pathogens: the importance of surface functionality to nanoparticle activity.

    PubMed

    Maguire-Boyle, Samuel J; Liga, Michael V; Li, Qilin; Barron, Andrew R

    2012-09-21

    A bi-functional nano-composite coating has been created on a porous Nomex® fabric support as a trap for aspirated virus contaminated water. Nomex® fabric was successively dip-coated in solutions containing cysteic acid functionalized alumina (alumoxane) nanoparticles and cysteic acid functionalized iron oxide (ferroxane) nanoparticles to form a nanoparticle coated Nomex® (NPN) fabric. From SEM and EDX the nanoparticle coating of the Nomex® fibers is uniform, continuous, and conformal. The NPN was used as a filter for aspirated bacteriophage MS2 viruses using end-on filtration. All measurements were repeated to give statistical reliability. The NPN fabrics show a large decrease as compared to Nomex® alone or alumoxane coated Nomex®. An increase in the ferroxane content results in an equivalent increase in virus retention. This suggests that it is the ferroxane that has an active role in deactivating and/or binding the virus. Heating the NPN to 160 °C results in the loss of cysteic acid functional groups (without loss of the iron nanoparticle's core structure) and the resulting fabric behaves similar to that of untreated Nomex®, showing that the surface functionalization of the nanoparticles is vital for the surface collapse of aspirated water droplets and the absorption and immobilization of the MS2 viruses. Thus, for virus immobilization, it is not sufficient to have iron oxide nanoparticles per se, but the surface functionality of a nanoparticle is vitally important in ensuring efficacy.

  17. The calcium-activated potassium channel KCa3.1 is an important modulator of hepatic injury.

    PubMed

    Sevelsted Møller, Linda; Fialla, Annette Dam; Schierwagen, Robert; Biagini, Matteo; Liedtke, Christian; Laleman, Wim; Klein, Sabine; Reul, Winfried; Koch Hansen, Lars; Rabjerg, Maj; Singh, Vikrant; Surra, Joaquin; Osada, Jesus; Reinehr, Roland; de Muckadell, Ove B Schaffalitzky; Köhler, Ralf; Trebicka, Jonel

    2016-01-01

    The calcium-activated potassium channel KCa3.1 controls different cellular processes such as proliferation and volume homeostasis. We investigated the role of KCa3.1 in experimental and human liver fibrosis. KCa3.1 gene expression was investigated in healthy and injured human and rodent liver. Effect of genetic depletion and pharmacological inhibition of KCa3.1 was evaluated in mice during carbon tetrachloride induced hepatic fibrogenesis. Transcription, protein expression and localisation of KCa3.1 was analysed by reverse transcription polymerase chain reaction, Western blot and immunohistochemistry. Hemodynamic effects of KCa3.1 inhibition were investigated in bile duct-ligated and carbon tetrachloride intoxicated rats. In vitro experiments were performed in rat hepatic stellate cells and hepatocytes. KCa3.1 expression was increased in rodent and human liver fibrosis and was predominantly observed in the hepatocytes. Inhibition of KCa3.1 aggravated liver fibrosis during carbon tetrachloride challenge but did not change hemodynamic parameters in portal hypertensive rats. In vitro, KCa3.1 inhibition leads to increased hepatocyte apoptosis and DNA damage, whereas proliferation of hepatic stellate cells was stimulated by KCa3.1 inhibition. Our data identifies KCa3.1 channels as important modulators in hepatocellular homeostasis. In contrast to previous studies in vitro and other tissues this channel appears to be anti-fibrotic and protective during liver injury. PMID:27354175

  18. The calcium-activated potassium channel KCa3.1 is an important modulator of hepatic injury

    PubMed Central

    Sevelsted Møller, Linda; Fialla, Annette Dam; Schierwagen, Robert; Biagini, Matteo; Liedtke, Christian; Laleman, Wim; Klein, Sabine; Reul, Winfried; Koch Hansen, Lars; Rabjerg, Maj; Singh, Vikrant; Surra, Joaquin; Osada, Jesus; Reinehr, Roland; de Muckadell, Ove B. Schaffalitzky; Köhler, Ralf; Trebicka, Jonel

    2016-01-01

    The calcium-activated potassium channel KCa3.1 controls different cellular processes such as proliferation and volume homeostasis. We investigated the role of KCa3.1 in experimental and human liver fibrosis. KCa3.1 gene expression was investigated in healthy and injured human and rodent liver. Effect of genetic depletion and pharmacological inhibition of KCa3.1 was evaluated in mice during carbon tetrachloride induced hepatic fibrogenesis. Transcription, protein expression and localisation of KCa3.1 was analysed by reverse transcription polymerase chain reaction, Western blot and immunohistochemistry. Hemodynamic effects of KCa3.1 inhibition were investigated in bile duct-ligated and carbon tetrachloride intoxicated rats. In vitro experiments were performed in rat hepatic stellate cells and hepatocytes. KCa3.1 expression was increased in rodent and human liver fibrosis and was predominantly observed in the hepatocytes. Inhibition of KCa3.1 aggravated liver fibrosis during carbon tetrachloride challenge but did not change hemodynamic parameters in portal hypertensive rats. In vitro, KCa3.1 inhibition leads to increased hepatocyte apoptosis and DNA damage, whereas proliferation of hepatic stellate cells was stimulated by KCa3.1 inhibition. Our data identifies KCa3.1 channels as important modulators in hepatocellular homeostasis. In contrast to previous studies in vitro and other tissues this channel appears to be anti-fibrotic and protective during liver injury. PMID:27354175

  19. Alumoxane/ferroxane nanoparticles for the removal of viral pathogens: the importance of surface functionality to nanoparticle activity

    NASA Astrophysics Data System (ADS)

    Maguire-Boyle, Samuel J.; Liga, Michael V.; Li, Qilin; Barron, Andrew R.

    2012-08-01

    A bi-functional nano-composite coating has been created on a porous Nomex® fabric support as a trap for aspirated virus contaminated water. Nomex® fabric was successively dip-coated in solutions containing cysteic acid functionalized alumina (alumoxane) nanoparticles and cysteic acid functionalized iron oxide (ferroxane) nanoparticles to form a nanoparticle coated Nomex® (NPN) fabric. From SEM and EDX the nanoparticle coating of the Nomex® fibers is uniform, continuous, and conformal. The NPN was used as a filter for aspirated bacteriophage MS2 viruses using end-on filtration. All measurements were repeated to give statistical reliability. The NPN fabrics show a large decrease as compared to Nomex® alone or alumoxane coated Nomex®. An increase in the ferroxane content results in an equivalent increase in virus retention. This suggests that it is the ferroxane that has an active role in deactivating and/or binding the virus. Heating the NPN to 160 °C results in the loss of cysteic acid functional groups (without loss of the iron nanoparticle's core structure) and the resulting fabric behaves similar to that of untreated Nomex®, showing that the surface functionalization of the nanoparticles is vital for the surface collapse of aspirated water droplets and the absorption and immobilization of the MS2 viruses. Thus, for virus immobilization, it is not sufficient to have iron oxide nanoparticles per se, but the surface functionality of a nanoparticle is vitally important in ensuring efficacy.A bi-functional nano-composite coating has been created on a porous Nomex® fabric support as a trap for aspirated virus contaminated water. Nomex® fabric was successively dip-coated in solutions containing cysteic acid functionalized alumina (alumoxane) nanoparticles and cysteic acid functionalized iron oxide (ferroxane) nanoparticles to form a nanoparticle coated Nomex® (NPN) fabric. From SEM and EDX the nanoparticle coating of the Nomex® fibers is uniform, continuous

  20. An Important Natural Genetic Resource of Oreochromis niloticus (Linnaeus, 1758) Threatened by Aquaculture Activities in Loboi Drainage, Kenya

    PubMed Central

    Ndiwa, Titus Chemandwa; Nyingi, Dorothy Wanja; Agnese, Jean-François

    2014-01-01

    The need to improve food security in Africa through culture of tilapias has led to transfer of different species from their natural ranges causing negative impacts on wild fish genetic resources. Loboi swamp in Kenya is fed by three hot springs: Lake Bogoria Hotel, Chelaba and Turtle Springs, hosting natural populations of Oreochromis niloticus. The present study aimed at better genetic characterization of these threatened populations. Partial mtDNA sequences of the D-loop region and variations at 16 microsatellite loci were assessed in the three hot spring populations and compared with three other natural populations of O. niloticus in the region. Results obtained indicated that the hot spring populations had mitochondrial and nuclear genetic variability similar to or higher than the large closely related populations. This may be attributed to the perennial nature of the hot springs, which do not depend on rainfall but rather receive permanent water supply from deep aquifers. The study also revealed that gene flow between the three different hot spring populations was sufficiently low thus allowing their differentiation. This differentiation was unexpected considering the very close proximity of the springs to each other. It is possible that the swamp creates a barrier to free movement of fish from one spring to the other thereby diminishing gene flow. Finally, the most surprising and worrying results were that the three hot spring populations are introgressed by mtDNA genes of O. leucostictus, while microsatellite analysis suggested that some nuclear genes may also have crossed the species barrier. It is very likely that the recent intensification of aquaculture activities in the Loboi drainage may be responsible for these introgressions. Taking into account the importance of these new genetic resources, protection and management actions of the Loboi swamp should be accorded top priority to prevent the loss of these spring populations. PMID:25222491

  1. An important natural genetic resource of Oreochromis niloticus (Linnaeus, 1758) threatened by aquaculture activities in Loboi drainage, Kenya.

    PubMed

    Ndiwa, Titus Chemandwa; Nyingi, Dorothy Wanja; Agnese, Jean-François

    2014-01-01

    The need to improve food security in Africa through culture of tilapias has led to transfer of different species from their natural ranges causing negative impacts on wild fish genetic resources. Loboi swamp in Kenya is fed by three hot springs: Lake Bogoria Hotel, Chelaba and Turtle Springs, hosting natural populations of Oreochromis niloticus. The present study aimed at better genetic characterization of these threatened populations. Partial mtDNA sequences of the D-loop region and variations at 16 microsatellite loci were assessed in the three hot spring populations and compared with three other natural populations of O. niloticus in the region. Results obtained indicated that the hot spring populations had mitochondrial and nuclear genetic variability similar to or higher than the large closely related populations. This may be attributed to the perennial nature of the hot springs, which do not depend on rainfall but rather receive permanent water supply from deep aquifers. The study also revealed that gene flow between the three different hot spring populations was sufficiently low thus allowing their differentiation. This differentiation was unexpected considering the very close proximity of the springs to each other. It is possible that the swamp creates a barrier to free movement of fish from one spring to the other thereby diminishing gene flow. Finally, the most surprising and worrying results were that the three hot spring populations are introgressed by mtDNA genes of O. leucostictus, while microsatellite analysis suggested that some nuclear genes may also have crossed the species barrier. It is very likely that the recent intensification of aquaculture activities in the Loboi drainage may be responsible for these introgressions. Taking into account the importance of these new genetic resources, protection and management actions of the Loboi swamp should be accorded top priority to prevent the loss of these spring populations.

  2. Alumoxane/ferroxane nanoparticles for the removal of viral pathogens: the importance of surface functionality to nanoparticle activity.

    PubMed

    Maguire-Boyle, Samuel J; Liga, Michael V; Li, Qilin; Barron, Andrew R

    2012-09-21

    A bi-functional nano-composite coating has been created on a porous Nomex® fabric support as a trap for aspirated virus contaminated water. Nomex® fabric was successively dip-coated in solutions containing cysteic acid functionalized alumina (alumoxane) nanoparticles and cysteic acid functionalized iron oxide (ferroxane) nanoparticles to form a nanoparticle coated Nomex® (NPN) fabric. From SEM and EDX the nanoparticle coating of the Nomex® fibers is uniform, continuous, and conformal. The NPN was used as a filter for aspirated bacteriophage MS2 viruses using end-on filtration. All measurements were repeated to give statistical reliability. The NPN fabrics show a large decrease as compared to Nomex® alone or alumoxane coated Nomex®. An increase in the ferroxane content results in an equivalent increase in virus retention. This suggests that it is the ferroxane that has an active role in deactivating and/or binding the virus. Heating the NPN to 160 °C results in the loss of cysteic acid functional groups (without loss of the iron nanoparticle's core structure) and the resulting fabric behaves similar to that of untreated Nomex®, showing that the surface functionalization of the nanoparticles is vital for the surface collapse of aspirated water droplets and the absorption and immobilization of the MS2 viruses. Thus, for virus immobilization, it is not sufficient to have iron oxide nanoparticles per se, but the surface functionality of a nanoparticle is vitally important in ensuring efficacy. PMID:22864822

  3. The 1911 M ~6.6 Calaveras earthquake: Source parameters and the role of static, viscoelastic, and dynamic coulomb stress changes imparted by the 1906 San Francisco earthquake

    USGS Publications Warehouse

    Doser, D.I.; Olsen, K.B.; Pollitz, F.F.; Stein, R.S.; Toda, S.

    2009-01-01

    The occurrence of a right-lateral strike-slip earthquake in 1911 is inconsistent with the calculated 0.2-2.5 bar static stress decrease imparted by the 1906 rupture at that location on the Calaveras fault, and 5 yr of calculated post-1906 viscoelastic rebound does little to reload the fault. We have used all available first-motion, body-wave, and surface-wave data to explore possible focal mechanisms for the 1911 earthquake. We find that the event was most likely a right-lateral strikeslip event on the Calaveras fault, larger than, but otherwise resembling, the 1984 Mw 6.1 Morgan Hill earthquake in roughly the same location. Unfortunately, we could recover no unambiguous surface fault offset or geodetic strain data to corroborate the seismic analysis despite an exhaustive archival search. We calculated the static and dynamic Coulomb stress changes for three 1906 source models to understand stress transfer to the 1911 site. In contrast to the static stress shadow, the peak dynamic Coulomb stress imparted by the 1906 rupture promoted failure at the site of the 1911 earthquake by 1.4-5.8 bar. Perhaps because the sample is small and the aftershocks are poorly located, we find no correlation of 1906 aftershock frequency or magnitude with the peak dynamic stress, although all aftershocks sustained a calculated dynamic stress of ???3 bar. Just 20 km to the south of the 1911 epicenter, we find that surface creep of the Calaveras fault at Hollister paused for ~17 yr after 1906, about the expected delay for the calculated static stress drop imparted by the 1906 earthquake when San Andreas fault postseismic creep and viscoelastic relaxation are included. Thus, the 1911 earthquake may have been promoted by the transient dynamic stresses, while Calaveras fault creep 20 km to the south appears to have been inhibited by the static stress changes.

  4. The Seeds of Learning: Young Children Develop Important Skills through Their Gardening Activities at a Midwestern Early Education Program

    ERIC Educational Resources Information Center

    Miller, Dana L.

    2007-01-01

    Using teachers as co-researchers to collect and analyze data, this case study explored preschool and kindergarteners' learning when they were engaged in hands-on activities in the garden and greenhouse areas of a model outdoor classroom. Key findings suggest that when young children are participating in garden and greenhouse activities they are:…

  5. 76 FR 4637 - Taking and Importing Marine Mammals; U.S. Navy's Atlantic Fleet Active Sonar Training

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-26

    ...&E) activities to be conducted within the Atlantic Fleet Active Sonar Training (AFAST) Study Area for... Integrated Comprehensive Management Program (ICMP) Plan, which is intended for use as a planning tool to... contacts listed here. The mailbox address for providing e-mail comments on the ICMP Plan is...

  6. The Importance of Physical Fitness versus Physical Activity for Coronary Artery Disease Risk Factors: A Cross-Sectional Analysis.

    ERIC Educational Resources Information Center

    Young, Deborah Rohm; Steinhardt, Mary A.

    1993-01-01

    This cross-sectional study examined relationships among physical fitness, physical activity, and risk factors for coronary artery disease (CAD) in male police officers. Data from screenings and physical fitness assessments indicated physical activity must be sufficient to influence fitness before obtaining statistically significant risk-reducing…

  7. Assessing the Importance of Social and Environmental Supports for Leisure-Time Physical Activity in a Unique Socio-Cultural and Geographical Context

    ERIC Educational Resources Information Center

    Gauthier, Alain P.; Lariviere, Michel; Pong, Raymond; Snelling, Susan J.; Young, Nancy L.

    2010-01-01

    Researchers have found important differences among population sub-groups when considering their "determinants of physical activity". The health of Francophones and Northern Ontario residents in Canada has been assessed as relatively poorer than the general population. Understanding the determinants of physical activity in these populations holds…

  8. The importance of activity-based methods in radiology and the technology that now makes this possible.

    PubMed

    Monge, Paul

    2006-01-01

    Activity-based methods serve as a dynamic process that has allowed many other industries to reduce and control their costs, increase productivity, and streamline their processes while improving product quality and service. The method could serve the healthcare industry in an equally beneficial way. Activity-based methods encompass both activity based costing (ABC) and activity-based management (ABM). ABC is a cost management approach that links resource consumption to activities that an enterprise performs, and then assigns those activities and their associated costs to customers, products, or product lines. ABM uses the resource assignments derived in ABC so that operation managers can improve their departmental processes and workflows. There are three fundamental problems with traditional cost systems. First, traditional systems fail to reflect the underlying diversity of work taking place within an enterprise. Second, it uses allocations that are, for the most part, arbitrary Single step allocations fail to reflect the real work-the activities being performed and the associate resources actually consumed. Third, they only provide a cost number that, standing alone, does not provide any guidance on how to improve performance by lowering cost or enhancing throughput. PMID:16783959

  9. Establishment of social club for adolescents living in urban slums: a potential tool to impart HIV/AIDS education.

    PubMed

    Deshpande, Kirti; Shivaswamy, M S; Diwan, Vishal; De Costa, Ayesha; Bhattacharya, Aruna; Mahadik, Vijay K

    2013-05-01

    An interventional study was carried out under a Government of Madhya Pradesh, India supported project with aims to describe establishment of a social club (a youth club), incorporating strategy of life-skill-based education and its potential as a tool for creating HIV/AIDS awareness among adolescents of age 15-19 years living in slums of Ujjain city. The social club activities included AIDS education by combined methods, vocational training and recreational activities for the adolescents. Establishment of social club was feasible with available local resources. Social club activities were need based, thus received good response from adolescents (94% had attendance above 75% in AIDS awareness sessions). Knowledge and attitude improved and misconceptions reduced significantly by AIDS education. Majority (86%) of the participants contributed as AIDS messengers in the community. The social clubs seem to have potential to improve HIV/AIDS awareness and empower adolescents to adopt healthy lifestyle through combination of teen friendly activities and atmosphere. PMID:24765690

  10. Nematicidal and antimicrobial activities of methanol extracts of 17 plants, of importance in ethnopharmacology, obtained from the Arabian Peninsula

    PubMed Central

    Al-Marby, Adel; Ejike, Chukwunonso ECC; Nasim, Muhammad Jawad; Awadh-Ali, Nasser A; Al-badani, Rwaida A; Alghamdi, Ghanem MA; Jacob, Claus

    2016-01-01

    Aim/Background: The development of resistance to synthetic drugs by target organisms is a major challenge facing medicine, yet locked within plants are phytochemicals used in herbal medicine (especially in the Arabian Peninsula) that may find application in this regard. In pursuit of unlocking these “hidden treasures,” the methanol extracts of leaves, aerial parts, fruits, and resins of 17 plants used in the Arabian Peninsula were screened for antimicrobial activities. Materials and Methods: The nematicidal, antibacterial, and antifungal activities were determined using appropriate assays. Steinernema feltiae, Staphylococcus carnosus, Escherichia coli, and Saccharomyces cerevisiae were used as test organisms. Concentrations of the extracts ranging from 0.5 to 20 mg/ml were tested and appropriate statistical tests performed on the data generated. Results: The results show that extracts from Solanum incanum, Chenopodium murale, Commiphora myrrha, Anthemis nobilis, and Achillea biebersteinii were the most active and had very high activities against two or more of the test organisms at low concentrations. Extracts of the leaves of S. incanum and resins of Ferula asafoetida were the most active nematicides, with significant activity at 0.5 mg/ml. Extracts of C. myrrha and C. murale had the most active antibacterial activity with inhibition zones of 12-15 mm and minimum inhibitory concentrations (MICs) of 2.5 mg/ml for both bacteria. Extracts of the leaves of A. biebersteinii were the most active fungicide, giving an MIC of 1.5 mg/ml. Conclusion: The results validate the use of these plants in ethnopharmacology, and open new vistas of opportunities for the development of cheap but effective agents that may be useful against infectious diseases. PMID:27104031

  11. The importance of culturally meaningful activity for health benefits among older Korean immigrant living in the United States.

    PubMed

    Kim, Junhyoung; Kim, May; Han, Areum; Chin, Seungtae

    2015-01-01

    Research indicates that participation in culturally meaningful activity is beneficial for immigrants' health and well-being, yet older Korean immigrants struggle with accepting new cultural perspectives, which can negatively affect their health and well-being. Using in-depth interviews, this study was designed to capture the value of culturally meaningful activities for health among older Korean immigrants. Three themes were identified: (a) improved psychological well-being, (b) enhanced positive emotions and feelings, and (c) social connections developed with others. The findings suggest that by engaging in various culturally meaningful activities, older Korean immigrants gain a sense of social, cultural, and psychological significance in life. This study also provided evidence that older Korean immigrants maintain and develop their cultural identity through culturally meaningful activities.

  12. The importance of culturally meaningful activity for health benefits among older Korean immigrant living in the United States

    PubMed Central

    Kim, Junhyoung; Kim, May; Han, Areum; Chin, Seungtae

    2015-01-01

    Research indicates that participation in culturally meaningful activity is beneficial for immigrants’ health and well-being, yet older Korean immigrants struggle with accepting new cultural perspectives, which can negatively affect their health and well-being. Using in-depth interviews, this study was designed to capture the value of culturally meaningful activities for health among older Korean immigrants. Three themes were identified: (a) improved psychological well-being, (b) enhanced positive emotions and feelings, and (c) social connections developed with others. The findings suggest that by engaging in various culturally meaningful activities, older Korean immigrants gain a sense of social, cultural, and psychological significance in life. This study also provided evidence that older Korean immigrants maintain and develop their cultural identity through culturally meaningful activities. PMID:26084272

  13. Developmental experiences during extracurricular activities and Australian adolescents' self-concept: particularly important for youth from disadvantaged schools.

    PubMed

    Blomfield, Corey J; Barber, Bonnie L

    2011-05-01

    Extracurricular activities provide adolescents with a number of positive personal and interpersonal developmental experiences. This study investigated whether developmental experiences that occurred during extracurricular activities were linked to a more positive self-concept for Australian adolescents, and whether this link was particularly salient for youth from disadvantaged schools. Adolescents (N = 1,504, 56% Female) from 26 diverse high schools across Western Australia were surveyed. The findings revealed that adolescents from low socio-economic status schools who participated in extracurricular activities had a more positive general self-worth and social self-concept than adolescents from similar socio-economic schools who did not participate in any extracurricular activities. Furthermore, the positive developmental experiences that occurred during extracurricular activities predicted a more positive general self-worth and social and academic self-concept, and this link was stronger for youth from low SES schools. These findings suggest that the developmental experiences afforded by extracurricular activities may foster positive adolescent development.

  14. Disruption of the immune-checkpoint VISTA gene imparts a proinflammatory phenotype with predisposition to the development of autoimmunity.

    PubMed

    Wang, Li; Le Mercier, Isabelle; Putra, Juan; Chen, Wenna; Liu, Jun; Schenk, Austin D; Nowak, Elizabeth C; Suriawinata, Arief A; Li, Jiannan; Noelle, Randolph J

    2014-10-14

    V domain-containing Ig suppressor of T-cell activation (VISTA) is a negative checkpoint regulator that suppresses T cell-mediated immune responses. Previous studies using a VISTA-neutralizing monoclonal antibody show that VISTA blockade enhances T-cell activation. The current study describes a comprehensive characterization of mice in which the gene for VISTA has been deleted. Despite the apparent normal hematopoietic development in young mice, VISTA genetic deficiency leads to a gradual accumulation of spontaneously activated T cells, accompanied by the production of a spectrum of inflammatory cytokines and chemokines. Enhanced T-cell responsiveness was also observed upon immunization with neoantigen. Despite the presence of multiorgan chronic inflammation, aged VISTA-deficient mice did not develop systemic or organ-specific autoimmune disease. Interbreeding of the VISTA-deficient mice with 2D2 T-cell receptor transgenic mice, which are predisposed to the development of experimental autoimmune encephalomyelitis, drastically enhanced disease incidence and intensity. Disease development is correlated with the increase in the activation of encephalitogenic T cells in the periphery and enhanced infiltration into the CNS. Taken together, our data suggest that VISTA is a negative checkpoint regulator whose loss of function lowers the threshold for T-cell activation, allowing for an enhanced proinflammatory phenotype and an increase in the frequency and intensity of autoimmunity under susceptible conditions.

  15. Implementing Policies to Enhance Physical Education and Physical Activity in Schools.

    PubMed

    Cooper, Kenneth H; Greenberg, Jayne D; Castelli, Darla M; Barton, Mitch; Martin, Scott B; Morrow, James R

    2016-06-01

    The purpose of this commentary is to provide an overview of national physical activity recommendations and policies (e.g., from the Institute of Medicine, National Physical Activity Plan, and Centers for Disease Control and Prevention) and to discuss how these important initiatives can be implemented in local schools. Successful policies are illustrated. Specific strategies and ideas are shared regarding how physical educators can assert themselves and impart their knowledge in an effort to build support for policy implementations that enhance the delivery of physical education and physical activity in their schools and communities. PMID:27100264

  16. Beyond Physical Activity: The Importance of Play and Nature-Based Play Spaces for Children's Health and Development.

    PubMed

    Herrington, Susan; Brussoni, Mariana

    2015-12-01

    The reduction of child obesity continues to be a challenge worldwide. Research indicates that playing outdoors, particularly in natural play spaces, boosts children's physical activity, potentially decreasing childhood obesity. We present evidence that natural play spaces also provide for more diverse forms of play for children of varying ages and competencies. This is crucial because play spaces designed expressly for physical activity may not increase physical activity among less active children. Moreover, when researchers only examine physical activity in play, they overlook the valuable contributions that play makes to other aspects of children's health and development. To enhance research on children and their play environments, we introduce the theory of play affordances. To assist in the creation of more natural play spaces, we describe the Seven Cs, an evidence-based approach for designing children's play spaces that promotes diverse play. We end with some preliminary insights from our current research using the Seven Cs to illustrate the connections between play, nature, and children's healthy development.

  17. Beyond Physical Activity: The Importance of Play and Nature-Based Play Spaces for Children's Health and Development.

    PubMed

    Herrington, Susan; Brussoni, Mariana

    2015-12-01

    The reduction of child obesity continues to be a challenge worldwide. Research indicates that playing outdoors, particularly in natural play spaces, boosts children's physical activity, potentially decreasing childhood obesity. We present evidence that natural play spaces also provide for more diverse forms of play for children of varying ages and competencies. This is crucial because play spaces designed expressly for physical activity may not increase physical activity among less active children. Moreover, when researchers only examine physical activity in play, they overlook the valuable contributions that play makes to other aspects of children's health and development. To enhance research on children and their play environments, we introduce the theory of play affordances. To assist in the creation of more natural play spaces, we describe the Seven Cs, an evidence-based approach for designing children's play spaces that promotes diverse play. We end with some preliminary insights from our current research using the Seven Cs to illustrate the connections between play, nature, and children's healthy development. PMID:26399254

  18. Federal environmental standards of potential importance to operations and activities at US Department of Energy sites. Draft

    SciTech Connect

    Fowler, K.M.; Bilyard, G.R.; Davidson, S.A.; Jonas, R.J.; Joseph, J.

    1993-06-01

    The US Department of Energy (DOE) is now engaged in a program of environmental restoration nationwide across its 45 sites. It is also bringing its facilities into compliance with environmental regulations, decontaminating and decommissioning unwanted facilities, and constructing new waste management facilities. One of the most difficult questions that DOE must face in successfully remediating its inactive waste sites, decontaminating and decommissioning its inactive facilities, and operating its waste management facilities is: ``What criteria and standards should be met?`` Acceptable standards or procedures for determining standards will assist DOE in its conduct of ongoing waste management and pending cleanup activities by helping to ensure that those activities are conducted in compliance with applicable laws and regulations and are accepted by the regulatory community and the public. This document reports on the second of three baseline activities that are being conducted as prerequisites to either the development of quantitative standards that could be used by DOE, or consistent procedures for developing such standards. The first and third baseline activities are also briefly discussed in conjunction with the second of the three activities.

  19. SAR studies on hydropentalene derivatives--Important core units of biologically active tetramic acid macrolactams and ptychanolides.

    PubMed

    Lutz, Vanessa; Mannchen, Fabian; Krebs, Michael; Park, Natja; Krüger, Claudia; Raja, Aruna; Sasse, Florenz; Baro, Angelika; Laschat, Sabine

    2014-07-01

    Structurally diverse bicyclo[3.3.0]octanes were prepared and tested for their biological activity. Both the antiproliferative activity and the results of phenotypic characterization varied with the substitution patterns. Two derivatives displayed high inhibitory (IC50 ≤3μM) activity against the L-929 cell line, but differed in their mode of action. A cluster analysis with impedance profiling data showed the two compounds in relationship to microtubule interfering compounds. In PtK2 cells treated with both derivatives a perturbing effect on the microtubular network was observed, whereas the actin cytoskeleton in incubated PtK2 cells was disturbed only by one compound. The effects on tubulin and actin polymerization could be confirmed by in vitro polymerization experiments.

  20. Molecular characterization of protease activity in Serratia sp. strain SCBI and its importance in cytotoxicity and virulence.

    PubMed

    Petersen, Lauren M; Tisa, Louis S

    2014-11-01

    A newly recognized Serratia species, termed South African Caenorhabditis briggsae isolate (SCBI), is both a mutualist of the nematode Caenorhabditis briggsae KT0001 and a pathogen of lepidopteran insects. Serratia sp. strain SCBI displays high proteolytic activity, and because secreted proteases are known virulence factors for many pathogens, the purpose of this study was to identify genes essential for extracellular protease activity in Serratia sp. strain SCBI and to determine what role proteases play in insect pathogenesis and cytotoxicity. A bank of 2,100 transposon mutants was generated, and six SCBI mutants with defective proteolytic activity were identified. These mutants were also defective in cytotoxicity. The mutants were found defective in genes encoding the following proteins: alkaline metalloprotease secretion protein AprE, a BglB family transcriptional antiterminator, an inosine/xanthosine triphosphatase, GidA, a methyl-accepting chemotaxis protein, and a PIN domain protein. Gene expression analysis on these six mutants showed significant downregulation in mRNA levels of several different types of predicted protease genes. In addition, transcriptome sequencing (RNA-seq) analysis provided insight into how inactivation of AprE, GidA, and a PIN domain protein influences motility and virulence, as well as protease activity. Using quantitative reverse transcription-PCR (qRT-PCR) to further characterize expression of predicted protease genes in wild-type Serratia sp. SCBI, the highest mRNA levels for the alkaline metalloprotease genes (termed prtA1 to prtA4) occurred following the death of an insect host, while two serine protease and two metalloprotease genes had their highest mRNA levels during active infection. Overall, these results indicate that proteolytic activity is essential for cytotoxicity in Serratia sp. SCBI and that its regulation appears to be highly complex.

  1. Molecular Characterization of Protease Activity in Serratia sp. Strain SCBI and Its Importance in Cytotoxicity and Virulence

    PubMed Central

    Petersen, Lauren M.

    2014-01-01

    A newly recognized Serratia species, termed South African Caenorhabditis briggsae isolate (SCBI), is both a mutualist of the nematode Caenorhabditis briggsae KT0001 and a pathogen of lepidopteran insects. Serratia sp. strain SCBI displays high proteolytic activity, and because secreted proteases are known virulence factors for many pathogens, the purpose of this study was to identify genes essential for extracellular protease activity in Serratia sp. strain SCBI and to determine what role proteases play in insect pathogenesis and cytotoxicity. A bank of 2,100 transposon mutants was generated, and six SCBI mutants with defective proteolytic activity were identified. These mutants were also defective in cytotoxicity. The mutants were found defective in genes encoding the following proteins: alkaline metalloprotease secretion protein AprE, a BglB family transcriptional antiterminator, an inosine/xanthosine triphosphatase, GidA, a methyl-accepting chemotaxis protein, and a PIN domain protein. Gene expression analysis on these six mutants showed significant downregulation in mRNA levels of several different types of predicted protease genes. In addition, transcriptome sequencing (RNA-seq) analysis provided insight into how inactivation of AprE, GidA, and a PIN domain protein influences motility and virulence, as well as protease activity. Using quantitative reverse transcription-PCR (qRT-PCR) to further characterize expression of predicted protease genes in wild-type Serratia sp. SCBI, the highest mRNA levels for the alkaline metalloprotease genes (termed prtA1 to prtA4) occurred following the death of an insect host, while two serine protease and two metalloprotease genes had their highest mRNA levels during active infection. Overall, these results indicate that proteolytic activity is essential for cytotoxicity in Serratia sp. SCBI and that its regulation appears to be highly complex. PMID:25182493

  2. The Importance of Motor Functional Levels from the Activity Limitation Perspective of ICF in Children with Cerebral Palsy

    ERIC Educational Resources Information Center

    Mutlu, Akmer

    2010-01-01

    Our purpose in this study was to evaluate performance and capacity as defined by Gross Motor Function Classification System (GMFCS) and Manual Ability Classification System (MACS) from the "activity limitation" perspective of International Classification of Functioning, Disability, and Health (ICF) and to investigate the relationship between the…

  3. Activation of the AMPK-ULK1 pathway plays an important role in autophagy during prion infection

    PubMed Central

    Fan, Xue-Yu; Tian, Chan; Wang, Hui; Xu, Yin; Ren, Ke; Zhang, Bao-Yun; Gao, Chen; Shi, Qi; Meng, Ge; Zhang, Lu-Bin; Zhao, Yang-Jing; Shao, Qi-Xiang; Dong, Xiao-Ping

    2015-01-01

    AMPK is a serine/threonine protein kinase that acts as a positive regulator of autophagy, by phosphorylating ULK1 at specific sites. A previous study demonstrated activation of the macroautophagic system in scrapie-infected experimental rodents and in certain human prion diseases, in which the essential negative regulator mTOR is severely inhibited. In this study, AMPK and ULK1 in the brains of hamsters infected with scrapie strain 263 K and in the scrapie-infected cell line SMB-S15 were analysed. The results showed an up-regulated trend of AMPK and AMPK-Thr172, ULK1 and ULK1-Ser555. Increases in brain AMPK and ULK1 occurred at an early stage of agent 263 K infection. The level of phosphorylated ULK1-Ser757 decreased during mid-infection and was only negligibly present at the terminal stage, a pattern that suggested a close relationship of the phosphorylated protein with altered endogenous mTOR. In addition, the level of LKB1 associated with AMPK activation was selectively increased at the early and middle stages of infection. Knockdown of endogenous ULK1 in SMB-S15 cells inhibited LC3 lipidation. These results showed that, in addition to the abolishment of the mTOR regulatory pathway, activation of the AMPK-ULK1 pathway during prion infection contributes to autophagy activation in prion-infected brain tissues. PMID:26423766

  4. 75 FR 5055 - Taking and Importing Marine Mammals; U.S. Navy's Atlantic Fleet Active Sonar Training (AFAST)

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-02-01

    ... to AFAST training, maintenance, and RDT&E became effective on January 22, 2009 (74 FR 4843, January... conducted within the AFAST Study Area under regulations issued on January 22, 2009 (74 FR 4843, January 27.... Navy's Atlantic Fleet Active Sonar Training (AFAST) AGENCY: National Marine Fisheries Service...

  5. The Serum Complement System: A Simplified Laboratory Exercise to Measure the Activity of an Important Component of the Immune System

    ERIC Educational Resources Information Center

    Inglis, Jordan E.; Radziwon, Kimberly A.; Maniero, Gregory D.

    2008-01-01

    The immune system is a vital physiological component that affords animals protection from disease and is composed of innate and adaptive mechanisms that rely on cellular and dissolved components. The serum complement system is a series of dissolved proteins that protect against a variety of pathogens. The activity of complement in serum can be…

  6. ERK1/2 signaling plays an important role in topoisomerase II poison-induced G2/M checkpoint activation.

    PubMed

    Kolb, Ryan H; Greer, Patrick M; Cao, Phu T; Cowan, Kenneth H; Yan, Ying

    2012-01-01

    Topo II poisons, which target topoisomerase II (topo II) to generate enzyme mediated DNA damage, have been commonly used for anti-cancer treatment. While clinical evidence demonstrate a capability of topo II poisons in inducing apoptosis in cancer cells, accumulating evidence also show that topo II poison treatment frequently results in cell cycle arrest in cancer cells, which was associated with subsequent resistance to these treatments. Results in this report indicate that treatment of MCF-7 and T47D breast cancer cells with topo II poisons resulted in an increased phosphorylation of extracellular signal-regulated kinase 1 and 2 (ERK1/2) and an subsequent induction of G2/M cell cycle arrest. Furthermore, inhibition of ERK1/2 activation using specific inhibitors markedly attenuated the topo II poison-induced G2/M arrest and diminished the topo II poison-induced activation of ATR and Chk1 kinases. Moreover, decreased expression of ATR by specific shRNA diminished topo II poison-induced G2/M arrest but had no effect on topo II poison-induced ERK1/2 activation. In contrast, inhibition of ERK1/2 signaling had little, if any, effect on topo II poison-induced ATM activation. In addition, ATM inhibition by either incubation of cells with ATM specific inhibitor or transfection of cells with ATM specific siRNA did not block topo II poison-induced G2/M arrest. Ultimately, inhibition of ERK1/2 signaling greatly enhanced topo II poison-induced apoptosis. These results implicate a critical role for ERK1/2 signaling in the activation of G2/M checkpoint response following topo II poison treatment, which protects cells from topo II poison-induced apoptosis.

  7. Regulated expression of platelet factor 4 in human monocytes--role of PARs as a quantitatively important monocyte activation pathway.

    PubMed

    Schaffner, Andreas; Rhyn, Petra; Schoedon, Gabriela; Schaer, Dominik J

    2005-07-01

    Human mononuclear phagocytes have recently been shown to express constitutively and even more so, upon stimulation with bacteria, fungi, lipopolysaccharide (LPS), zymosan, or thrombin platelet basic protein (PBP). This CXC chemokine as well as platelet factor 4 (PF4), which is located genomically at a short distance from the PBP, were previously considered to be specific markers for the megakaryocyte cell lineage. Both chemokines have signaling and antimicrobial activity. In the present studies, transcriptional and expressional regulation of PF4 and related chemokines was studied in human monocytes. As shown by quantitative mRNA analysis, Western blots, radioimmunoprecipitation of cell extracts, and immunofluorescence and quantitatively with enzyme-linked immunosorbent assay, human monocytes express PF4 in the same order of magnitude as the known, regulated CXC chemokine interleukin (IL)-8. Expression of PF4 is up-regulated at the mRNA and protein level by thrombin and mediated by proteinase-activated receptors (PARs), resulting in a 32- to 128-fold higher mRNA level and leading to an up-to-sixfold increase of the peptide concentration in monocyte culture supernatants. Thrombin and the synthetic ligand of PAR-1 and PAR-2, SFLLRN, also induced comparable increases in the levels of mRNA for PBP, IL-8, regulated on activation, normal T expressed and secreted (RANTES), monocyte chemoattractant protein-1, and macrophage-inflammatory protein-1alpha and increased synthesis of these chemokines as shown by immunofluorescence or a quantitative immunobead-based method. The induction of increased mRNA levels for all chemokines by SFLLRN was unsurpassed by LPS, zymosan, interferon-gamma (IFN-gamma), tumor necrosis factor alpha (TNF-alpha), and IL-1. Activation of monocytes through PARs represents an alternate activation mechanism, independent from IFN-gamma, TNF-alpha, or other signaling pathways. PMID:15788441

  8. Importance of the CCR5-CCL5 axis for mucosal Trypanosoma cruzi protection and B cell activation.

    PubMed

    Sullivan, Nicole L; Eickhoff, Christopher S; Zhang, Xiuli; Giddings, Olivia K; Lane, Thomas E; Hoft, Daniel F

    2011-08-01

    Trypanosoma cruzi is an intracellular parasite and the causative agent of Chagas disease. Previous work has shown that the chemokine receptor CCR5 plays a role in systemic T. cruzi protection. We evaluated the importance of CCR5 and CCL5 for mucosal protection against natural oral and conjunctival T. cruzi challenges. T. cruzi-immune CCR5(-/-) and wild-type C57BL/6 mice were generated by repeated infectious challenges with T. cruzi. CCR5(-/-) and wild-type mice developed equivalent levels of cellular, humoral, and protective mucosal responses. However, CCR5(-/-)-immune mice produced increased levels of CCL5 in protected gastric tissues, suggesting compensatory signaling through additional receptors. Neutralization of CCL5 in CCR5(-/-)-immune mice resulted in decreased mucosal inflammatory responses, reduced T. cruzi-specific Ab-secreting cells, and significantly less mucosal T. cruzi protection, confirming an important role for CCL5 in optimal immune control of T. cruzi replication at the point of initial mucosal invasion. To investigate further the mechanism responsible for mucosal protection mediated by CCL5-CCR5 signaling, we evaluated the effects of CCL5 on B cells. CCL5 enhanced proliferation and IgM secretion in highly purified B cells triggered by suboptimal doses of LPS. In addition, neutralization of endogenous CCL5 inhibited B cell proliferation and IgM secretion during stimulation of highly purified B cells, indicating that B cell production of CCL5 has important autocrine effects. These findings demonstrate direct effects of CCL5 on B cells, with significant implications for the development of mucosal adjuvants, and further suggest that CCL5 may be important as a general B cell coactivator.

  9. 2-(Nitroaryl)benzothiazole and benzoxazole derivatives as fluorogenic substrates for the detection of nitroreductase activity in clinically important microorganisms.

    PubMed

    Cellier, Marie; Gignoux, Amandine; James, Arthur L; Orenga, Sylvain; Perry, John D; Robinson, Shaun N; Stanforth, Stephen P; Turnbull, Graeme

    2015-12-15

    A series of carboxy-substituted 2-(nitroaryl)benzothiazole derivatives and carboxy-substituted 2-(nitroaryl)benzoxazole derivatives were prepared and evaluated as potential nitroreductase substrates for the purpose of detecting clinically important microorganisms. Several of the substrates produced highly fluorescent colonies with the majority of a panel of 10 Gram-negative bacteria and also with two of a panel of 8 Gram-positive bacteria.

  10. Important role of surface fluoride in nitrogen-doped TiO2 nanoparticles with visible light photocatalytic activity.

    PubMed

    Brauer, Jonathan I; Szulczewski, Greg

    2014-12-11

    Nitrogen-doped TiO2 nanoparticles have been synthesized using sol-gel methods and subsequently fluorinated at room temperature by aging in acidic solutions of NaF. The nanoparticles were characterized by X-ray photoelectron spectroscopy, X-ray diffraction, UV-vis, and IR diffuse reflectance spectroscopy. After aging at room temperature in NaF solutions, the Ti-OH groups on the surface of the TiO2 nanoparticles were replaced by Ti-F bonds, which resulted in a decrease of the point of zero charge from pH 5.4 to 2.8. Most importantly, the nitrogen dopants were retained after the fluorination process, and the amorphous nanoparticles were partially converted into the anatase phase. Annealing the photocatalysts resulted in a decrease of both the nitrogen and fluoride atomic concentration. Diffuse reflectance spectra show an increase in absorbance above 400 nm after annealing the F,N-doped TiO2, which suggests the formation of color centers. The photoactivity of the F,N-doped and N-doped TiO2 catalysts were evaluated by monitoring by the decolorization of methylene blue with visible light. Mass spectrometric analysis revealed that methylene blue undergoes successive demethylation, and more importantly, the rate of decolorization depends on the fluoride concentration. These results show the importance of a two-step synthesis method to independently control the nitrogen and fluoride concentration.

  11. [Correlation between clinical parameters and quantitative analysis of inflammatory infiltrate. Importance in the diagnosis of active periodontal disease].

    PubMed

    Mendieta Fiter, C

    1989-05-01

    Recent studies have indicated that the destruction in chronic periodontal disease occurs in relatively short periods of time which are followed of prolonged periods of inactivity. These bursts of activity are characterized by an increase in the inflammatory reaction. It has been the purpose of this paper to study the correlation between the clinical measurements of periodontal disease and the histomorphometric quantification of areas of dense inflammatory infiltrate. The results of this study show that the clinical parameters that measure gingival inflammation or loss of periodontal attachment are useful to distinguish pathology from normal (p less than 0.003), but lack sensitivity to detect burst of periodontal disease activity (p greater than 0.05). PMID:2489938

  12. [Correlation between clinical parameters and quantitative analysis of inflammatory infiltrate. Importance in the diagnosis of active periodontal disease].

    PubMed

    Mendieta Fiter, C

    1989-05-01

    Recent studies have indicated that the destruction in chronic periodontal disease occurs in relatively short periods of time which are followed of prolonged periods of inactivity. These bursts of activity are characterized by an increase in the inflammatory reaction. It has been the purpose of this paper to study the correlation between the clinical measurements of periodontal disease and the histomorphometric quantification of areas of dense inflammatory infiltrate. The results of this study show that the clinical parameters that measure gingival inflammation or loss of periodontal attachment are useful to distinguish pathology from normal (p less than 0.003), but lack sensitivity to detect burst of periodontal disease activity (p greater than 0.05).

  13. Metals content of Glossoscolex paulistus extracellular hemoglobin: Its peroxidase activity and the importance of these ions in the protein stability.

    PubMed

    Caruso, Celia S; Biazin, Ezer; Carvalho, Francisco A O; Tabak, Marcel; Bachega, José F R

    2016-08-01

    In this work we investigate the presence of divalent cations bound to the Glossoscolex paulistus (HbGp) hemoglobin and their effect over the protein stability and the peroxidase (POD) activity. Atomic absorption studies show that the HbGp iron content is consistent with the presence of 144 ions per protein. Moreover, using iron as a reference, the content of calcium was estimated as 30±4 ions per protein, independently of the EDTA pre-treatment or not prior to the acidic treatment performed in the protein digestion. The zinc content was 14±2 ions in the absence of EDTA pre-treatment, and 3±1 ions per protein in the presence of EDTA pre-treatment, implying the presence of one zinc ion per protomer (1/12 of the whole molecule). Finally, the copper concentration is negligible. Different from the vertebrate hemoglobins, where the effectors are usually organic anions, the hexagonal bilayer hemoglobins have as effectors inorganic cations that increase the oxygen affinity and stabilize the structure. Previous studies have suggested that the presence of divalent cations, such as copper and zinc, is related to the different types of antioxidant enzymatic activities as the superoxide dismutase (SOD) activity shown by giant hemoglobin from Lumbricus terrestris (HbLt). Recently, studies on HbGp crystal structure have confirmed the presence of Zn(2+) and Ca(2+) binding sites. The Ca(2+) sites are similar as observed in the HbLt crystal structure. Otherwise, the Zn(2+) sites have no relation with those observed in Cu/Zn SODs. Our peroxidase assays with guaiacol confirm the POD activity and the effect of the zinc ions for HbGp. Our present results on HbGp metal content and their stability effects is the first step to understand the role of these cations in HbGp function in the future. PMID:27221949

  14. Metabolism of hydroxypyruvate in a mutant of barley lacking NADH-dependent hydroxypyruvate reductase, an important photorespiratory enzyme activity

    SciTech Connect

    Murray, A.J.S.; Blackwell, R.D.; Lea, P.J. )

    1989-09-01

    A mutant of barley (Hordeum vulgare L.), LaPr 88/29, deficient in NADH-dependent hydroxypyruvate reductase (HPR) activity has been isolated. The activities of both NADH (5%) and NADPH-dependent (19%) HPR were severely reduced in this mutant compared to the wild type. Although lacking an enzyme in the main carbon pathway of photorespiration, this mutant was capable of CO{sub 2} fixation rates equivalent to 75% of that of the wild type, in normal atmospheres and 50% O{sub 2}. There also appeared to be little disruption to the photorespiratory metabolism as ammonia release, CO{sub 2} efflux and {sup 14}CO{sub 2} release from L-(U-{sup 14}C)serine feeding were similar in both mutant and wild-type leaves. When leaves of LaPr 88/29 were fed either ({sup 14}C)serine or {sup 14}CO{sub 2}, the accumulation of radioactivity was in serine and not in hydroxypyruvate, although the mutant was still able to metabolize over 25% of the supplied ({sup 14}C)serine into sucrose. After 3 hours in air the soluble amino acid pool was almost totally dominated by serine and glycine. LaPr 88/29 has also been used to show that NADH-glyoxylate reductase and NADH-HPR are probably not catalyzed by the same enzyme in barley and that over 80% of the NADPH-dependent HPR activity is due to the NADH-dependent enzyme. We also suggest that the alternative NADPH activity can metabolize a proportion, but not all, of the hydroxypyruvate produced during photorespiration and may thus form a useful backup to the NADH-dependent enzyme under conditions of maximal photorespiration.

  15. Low resolution solution structure of HAMLET and the importance of its alpha-domains in tumoricidal activity.

    PubMed

    Ho, C S James; Rydstrom, Anna; Manimekalai, Malathy Sony Subramanian; Svanborg, Catharina; Grüber, Gerhard

    2012-01-01

    HAMLET (Human Alpha-lactalbumin Made LEthal to Tumor cells) is the first member in a new family of protein-lipid complexes with broad tumoricidal activity. Elucidating the molecular structure and the domains crucial for HAMLET formation is fundamental for understanding its tumoricidal function. Here we present the low-resolution solution structure of the complex of oleic acid bound HAMLET, derived from small angle X-ray scattering data. HAMLET shows a two-domain conformation with a large globular domain and an extended part of about 2.22 nm in length and 1.29 nm width. The structure has been superimposed into the related crystallographic structure of human α-lactalbumin, revealing that the major part of α-lactalbumin accommodates well in the shape of HAMLET. However, the C-terminal residues from L105 to L123 of the crystal structure of the human α-lactalbumin do not fit well into the HAMLET structure, resulting in an extended conformation in HAMLET, proposed to be required to form the tumoricidal active HAMLET complex with oleic acid. Consistent with this low resolution structure, we identified biologically active peptide epitopes in the globular as well as the extended domains of HAMLET. Peptides covering the alpha1 and alpha2 domains of the protein triggered rapid ion fluxes in the presence of sodium oleate and were internalized by tumor cells, causing rapid and sustained changes in cell morphology. The alpha peptide-oleate bound forms also triggered tumor cell death with comparable efficiency as HAMLET. In addition, shorter peptides corresponding to those domains are biologically active. These findings provide novel insights into the structural prerequisites for the dramatic effects of HAMLET on tumor cells.

  16. Importance of surface carbide formation on the activity and selectivity of Pd surfaces in the selective hydrogenation of acetylene

    NASA Astrophysics Data System (ADS)

    Yang, Bo; Burch, Robbie; Hardacre, Christopher; Hu, P.; Hughes, Philip

    2016-04-01

    A recent experimental investigation (Kim et al. J. Catal. 306 (2013) 146-154) on the selective hydrogenation of acetylene over Pd nanoparticles with different shapes concluded that Pd(100) showed higher activity and selectivity than Pd(111) for acetylene hydrogenation. However, our recent density functional calculations (Yang et al. J. Catal. 305 (2013) 264-276) observed that the clean Pd(111) surface should result in higher activity and ethylene selectivity compared with the clean Pd(100) surface for acetylene hydrogenation. In the current work, using density functional theory calculations, we find that Pd(100) in the carbide form gives rise to higher activity and selectivity than Pd(111) carbide. These results indicate that the catalyst surface is most likely in the carbide form under the experimental reaction conditions. Furthermore, the adsorption energies of hydrogen atoms as a function of the hydrogen coverage at the surface and subsurface sites over Pd(100) are compared with those over Pd(111), and it is found that the adsorption of hydrogen atoms is always less favoured on Pd(100) over the whole coverage range. This suggests that the Pd(100) hydride surface will be less stable than the Pd(111) hydride surface, which is also in accordance with the experimental results reported.

  17. Endospores of B subtilis are pyrogenic and activate Mono Mac 6 cells: importance of the CD14 receptor.

    PubMed

    Moesby, Lise; Hansen, Eirk W; Christensen, Jens D; Tommerup, Lene; Nielsen, Christina

    2003-07-01

    The monocytic cell line Mono Mac 6 is sensitive to pyrogens and interleukin-6 secretion is induced after exposure to pyrogens. The aim of this study is to examine the pyrogenic activity and the interleukin-6-inducing capacity of the Gram-positive B. subtilis bacteria, endospores and isolated cell wall components. Furthermore the involvement of CD14 in activation of interleukin-6 release is investigated. All test substances are pyrogenic in the rabbit pyrogen test. The test substance is incubated with monocytic cells (Mono Mac 6) for 24 h and the secreted interleukin-6 is determined in a sandwich immunoassay. B. subtilis bacteria and endospores induce interleukin-6 in a dose-dependent manner. Endospores are less potent than bacteria. Lipoteichoic acid (LTA) isolated from B. subtilis induces interleukin-6 in a dose-dependent manner, whereas muramyl dipeptide (MDP) is unable to induce interleukin-6. Lipopolysaccharides (LPS) dose-dependently induce interleukin-6 release, but the curve differs from that of LTA both in shape and offset. The interleukin-6 secretion induced by LPS, LTA and B. subtilis bacteria can be blocked by 73-85% by an antibody directed against CD14, whereas the antibody only blocks 25% of B. subtilis endospores-induced interleukin-6 release. The results might indicate that B. subtilis endospores use an additional pathway to CD14 to activate mononuclear cells.

  18. Understanding the Importance of Context: A Qualitative Study of a Location-Based Exergame to Enhance School Childrens Physical Activity.

    PubMed

    Robertson, Judy; Jepson, Ruth; Macvean, Andrew; Gray, Stuart

    2016-01-01

    Many public health interventions are less effective than expected in 'real life settings', yet little work is undertaken to understand the reasons why. The effectiveness of complex public health interventions can often be traced back to a robust programme theory (how and why an intervention brings about a change in outcome(s)) and assumptions that are made about the context in which it is implemented. Understanding whether effectiveness (or lack thereof) is due to the intervention or the context is hugely helpful in decisions about whether to a) modify the intervention; b) modify the context; c) stop providing the intervention. Exergames-also known as Active Video Games or AVGS-are video games which use the player's bodily movements as input and have potential to increase physical activity in children. However, the results of a recent pilot randomised controlled trial (RCT) of a location-based exergame (FitQuest) in a school setting were inconclusive; no significant effect was detected for any of the outcome measures. The aim of this study was to explore whether the programme theory for FitQuest was correct with respect to how and why it would change children's perceptions of physical activity (PA) and exercise self-efficacy in the school setting. A further aim was to investigate the features of the school setting (context) that may impact on FitQuest's implementation and effectiveness. Qualitative data (gathered during the RCT) were gathered from interviews with teachers and children, and observation of sessions using FitQuest. Thematic analysis indicated that whilst children enjoyed playing the game, engaged with goal setting within the game context and undertook low to vigorous physical activity, there were significant contextual factors that prevented it from being played as often as intended. These included environmental factors (e.g. size of the playground), school factors (cancellations due to other activities), school technology policy (rules relating to

  19. Understanding the Importance of Context: A Qualitative Study of a Location-Based Exergame to Enhance School Childrens Physical Activity

    PubMed Central

    Jepson, Ruth; Macvean, Andrew; Gray, Stuart

    2016-01-01

    Many public health interventions are less effective than expected in ‘real life settings’, yet little work is undertaken to understand the reasons why. The effectiveness of complex public health interventions can often be traced back to a robust programme theory (how and why an intervention brings about a change in outcome(s)) and assumptions that are made about the context in which it is implemented. Understanding whether effectiveness (or lack thereof) is due to the intervention or the context is hugely helpful in decisions about whether to a) modify the intervention; b) modify the context; c) stop providing the intervention. Exergames–also known as Active Video Games or AVGS–are video games which use the player's bodily movements as input and have potential to increase physical activity in children. However, the results of a recent pilot randomised controlled trial (RCT) of a location-based exergame (FitQuest) in a school setting were inconclusive; no significant effect was detected for any of the outcome measures. The aim of this study was to explore whether the programme theory for FitQuest was correct with respect to how and why it would change children’s perceptions of physical activity (PA) and exercise self-efficacy in the school setting. A further aim was to investigate the features of the school setting (context) that may impact on FitQuest’s implementation and effectiveness. Qualitative data (gathered during the RCT) were gathered from interviews with teachers and children, and observation of sessions using FitQuest. Thematic analysis indicated that whilst children enjoyed playing the game, engaged with goal setting within the game context and undertook low to vigorous physical activity, there were significant contextual factors that prevented it from being played as often as intended. These included environmental factors (e.g. size of the playground), school factors (cancellations due to other activities), school technology policy (rules

  20. Understanding the Importance of Context: A Qualitative Study of a Location-Based Exergame to Enhance School Childrens Physical Activity.

    PubMed

    Robertson, Judy; Jepson, Ruth; Macvean, Andrew; Gray, Stuart

    2016-01-01

    Many public health interventions are less effective than expected in 'real life settings', yet little work is undertaken to understand the reasons why. The effectiveness of complex public health interventions can often be traced back to a robust programme theory (how and why an intervention brings about a change in outcome(s)) and assumptions that are made about the context in which it is implemented. Understanding whether effectiveness (or lack thereof) is due to the intervention or the context is hugely helpful in decisions about whether to a) modify the intervention; b) modify the context; c) stop providing the intervention. Exergames-also known as Active Video Games or AVGS-are video games which use the player's bodily movements as input and have potential to increase physical activity in children. However, the results of a recent pilot randomised controlled trial (RCT) of a location-based exergame (FitQuest) in a school setting were inconclusive; no significant effect was detected for any of the outcome measures. The aim of this study was to explore whether the programme theory for FitQuest was correct with respect to how and why it would change children's perceptions of physical activity (PA) and exercise self-efficacy in the school setting. A further aim was to investigate the features of the school setting (context) that may impact on FitQuest's implementation and effectiveness. Qualitative data (gathered during the RCT) were gathered from interviews with teachers and children, and observation of sessions using FitQuest. Thematic analysis indicated that whilst children enjoyed playing the game, engaged with goal setting within the game context and undertook low to vigorous physical activity, there were significant contextual factors that prevented it from being played as often as intended. These included environmental factors (e.g. size of the playground), school factors (cancellations due to other activities), school technology policy (rules relating to

  1. Decoding the membrane activity of the cyclotide kalata B1: the importance of phosphatidylethanolamine phospholipids and lipid organization on hemolytic and anti-HIV activities.

    PubMed

    Henriques, Sónia Troeira; Huang, Yen-Hua; Rosengren, K Johan; Franquelim, Henri G; Carvalho, Filomena A; Johnson, Adam; Sonza, Secondo; Tachedjian, Gilda; Castanho, Miguel A R B; Daly, Norelle L; Craik, David J

    2011-07-01

    Cyclotides, a large family of cyclic peptides from plants, have a broad range of biological activities, including insecticidal, cytotoxic, and anti-HIV activities. In all of these activities, cell membranes seem likely to be the primary target for cyclotides. However, the mechanistic role of lipid membranes in the activity of cyclotides remains unclear. To determine the role of lipid organization in the activity of the prototypic cyclotide, kalata B1 (kB1), and synthetic analogs, their bioactivities and affinities for model membranes were evaluated. We found that the bioactivity of kB1 is dependent on the lipid composition of target cell membranes. In particular, the activity of kB1 requires specific interactions with phospholipids containing phosphatidylethanolamine (PE) headgroups but is further modulated by nonspecific peptide-lipid hydrophobic interactions, which are favored in raft-like membranes. Negatively charged phospholipids do not favor high kB1 affinity. This lipid selectivity explains trends in antimicrobial and hemolytic activities of kB1; it does not target bacterial cell walls, which are negatively charged and lacking PE-phospholipids but can insert in the membranes of red blood cells, which have a low PE content and raft domains in their outer layer. We further show that the anti-HIV activity of kB1 is the result of its ability to target and disrupt the membranes of HIV particles, which are raft-like membranes very rich in PE-phospholipids.

  2. Heterologous Expression of Two Jatropha Aquaporins Imparts Drought and Salt Tolerance and Improves Seed Viability in Transgenic Arabidopsis thaliana.

    PubMed

    Khan, Kasim; Agarwal, Pallavi; Shanware, Arti; Sane, Vidhu Aniruddha

    2015-01-01

    Drought and high salinity are environmental conditions that cause adverse effects on the growth and productivity of crops. Aquaporins are small integral membrane proteins that belong to the family of the major intrinsic proteins (MIPs), with members in animals, plants and microbes, where they facilitate the transport of water and/or small neutral solutes thereby affecting water balance. In this study we characterized two aquaporin genes namely, plasma membrane intrinsic protein (PIP2;7) and tonoplast intrinsic protein TIP1;3 from Jatropha curcas that are localised to the plasma membrane and vacuole respectively. Transgenic Arabidopsis thaliana lines over-expressing JcPIP2;7 and JcTIP1;3 under a constitutive promoter show improved germination under high salt and mannitol compared to control seeds. These transgenic plants also show increased root length under abiotic stress conditions compared to wild type Col-0 plants. Transgenic lines exposed to drought conditions by withholding water for 20 days, were able to withstand water stress and attained normal growth after re-watering unlike control plants which could not survive. Transgenic lines also had better seed yield than control under salt stress. Importantly, seed viability of transgenic plants grown under high salt concentration was 35%-45% compared to less than 5% for control seeds obtained from plants growing under salt stress. The effect of JcPIP2;7 and JcTIP1;3 on improving germination and seed viability in drought and salinity make these important candidates for genetic manipulation of plants for growth in saline soils. PMID:26067295

  3. Heterologous Expression of Two Jatropha Aquaporins Imparts Drought and Salt Tolerance and Improves Seed Viability in Transgenic Arabidopsis thaliana

    PubMed Central

    Khan, Kasim; Agarwal, Pallavi; Shanware, Arti; Sane, Vidhu Aniruddha

    2015-01-01

    Drought and high salinity are environmental conditions that cause adverse effects on the growth and productivity of crops. Aquaporins are small integral membrane proteins that belong to the family of the major intrinsic proteins (MIPs), with members in animals, plants and microbes, where they facilitate the transport of water and/or small neutral solutes thereby affecting water balance. In this study we characterized two aquaporin genes namely, plasma membrane intrinsic protein (PIP2;7) and tonoplast intrinsic protein TIP1;3 from Jatropha curcas that are localised to the plasma membrane and vacuole respectively. Transgenic Arabidopsis thaliana lines over-expressing JcPIP2;7 and JcTIP1;3 under a constitutive promoter show improved germination under high salt and mannitol compared to control seeds. These transgenic plants also show increased root length under abiotic stress conditions compared to wild type Col-0 plants. Transgenic lines exposed to drought conditions by withholding water for 20 days, were able to withstand water stress and attained normal growth after re-watering unlike control plants which could not survive. Transgenic lines also had better seed yield than control under salt stress. Importantly, seed viability of transgenic plants grown under high salt concentration was 35%-45% compared to less than 5% for control seeds obtained from plants growing under salt stress. The effect of JcPIP2;7 and JcTIP1;3 on improving germination and seed viability in drought and salinity make these important candidates for genetic manipulation of plants for growth in saline soils. PMID:26067295

  4. Heterologous Expression of Two Jatropha Aquaporins Imparts Drought and Salt Tolerance and Improves Seed Viability in Transgenic Arabidopsis thaliana.

    PubMed

    Khan, Kasim; Agarwal, Pallavi; Shanware, Arti; Sane, Vidhu Aniruddha

    2015-01-01

    Drought and high salinity are environmental conditions that cause adverse effects on the growth and productivity of crops. Aquaporins are small integral membrane proteins that belong to the family of the major intrinsic proteins (MIPs), with members in animals, plants and microbes, where they facilitate the transport of water and/or small neutral solutes thereby affecting water balance. In this study we characterized two aquaporin genes namely, plasma membrane intrinsic protein (PIP2;7) and tonoplast intrinsic protein TIP1;3 from Jatropha curcas that are localised to the plasma membrane and vacuole respectively. Transgenic Arabidopsis thaliana lines over-expressing JcPIP2;7 and JcTIP1;3 under a constitutive promoter show improved germination under high salt and mannitol compared to control seeds. These transgenic plants also show increased root length under abiotic stress conditions compared to wild type Col-0 plants. Transgenic lines exposed to drought conditions by withholding water for 20 days, were able to withstand water stress and attained normal growth after re-watering unlike control plants which could not survive. Transgenic lines also had better seed yield than control under salt stress. Importantly, seed viability of transgenic plants grown under high salt concentration was 35%-45% compared to less than 5% for control seeds obtained from plants growing under salt stress. The effect of JcPIP2;7 and JcTIP1;3 on improving germination and seed viability in drought and salinity make these important candidates for genetic manipulation of plants for growth in saline soils.

  5. The Importance of Establishing and Maintaining Continuity of Knowledge during 21st Century Nuclear Fuel Cycle Activities

    SciTech Connect

    Pickett, Chris A; Rowe, Nathan C; Younkin, James R; Wishard, Bernard; Bean, Robert; Blair, Dianna; Lawson, Ray; Weeks, George; Tolk, Keith

    2012-01-01

    During this century, the entire nuclear fuel cycle will expand and become increasingly more global, taxing both the resources and capabilities of the International Atomic Energy Agency (IAEA) to maintain an effective Continuity of Knowledge (CoK) and its ability to provide timely detection of diversion. Uranium that currently is mined and milled in one country will be converted, enriched, and fabricated into fuel for reactors in an expanding set of new countries. This expansion will make it harder to guarantee that regional activities stay regional and that diversion detection is timely unless new and sustainable tools are developed to improve inspector effectiveness. To deal with this emerging reality, the IAEA must increase its use of unattended monitoring and employ new tools and methods that enhance CoK during all phases of the fuel cycle. This approach will help provide useful information to aid in detecting undeclared activities and create opportunities for timely and appropriate responses to events well before they enter phases of greater concern (e.g., enrichment). The systems that maintain CoK of safeguarded assets rely on containment and surveillance (C/S) technologies. The 21st century fuel cycle will require increased use of these technologies and systems, plus greater implementation of unattended systems that can securely collect data when inspectors are not present.

  6. Sports Facilities, Shopping Centers or Homes: What Locations are Important for Adults' Physical Activity? A Cross-Sectional Study.

    PubMed

    Jansen, Marijke; Ettema, Dick; Pierik, Frank; Dijst, Martin

    2016-03-04

    Physical activity (PA) is influenced by the built environment. However, little is known about the types of built environment where adults spend their time, and at what levels of PA they engage in those environments. Understanding the effect of the built environment on PA requires insight into PA behavior at different types of locations (e.g., home, work, shopping centers, and sports facilities). Therefore, this study describes where adults aged 45-65 years were active with moderate-to-vigorous intensity (MVPA), and examines associations of socio-demographic factors and neighborhood with MVPA at these locations. Participants' (N = 308) PA was measured for seven days using accelerometers and GPS-devices. Adults spent most minutes of MVPA at home and work. Highest MVPA-ratios of total time spent at a location were achieved in sports facilities and during transport. Neighborhood characteristics and socio-demographic factors such as work status, health status and household structure, had significant effects on MVPA at various locations and on total MVPA. Understanding PA behavior at various locations may provide insights that allow professionals in different domains (e.g., health, landscaping, urban planning) to develop strategies to stimulate PA.

  7. The Influence of Organized Physical Activity (including Gymnastics) on Young Adult Skeletal Traits: Is Maturity Phase Important?

    PubMed Central

    Bernardoni, Brittney; Scerpella, Tamara A.; Rosenbaum, Paula F.; Kanaley, Jill A.; Raab, Lindsay N.; Li, Quefeng; Wang, Sijian; Dowthwaite, Jodi N.

    2015-01-01

    We prospectively evaluated adolescent organized physical activity (PA) as a factor in adult female bone traits. Annual DXA scans accompanied semi-annual records of anthropometry, maturity and PA for 42 participants in this preliminary analysis (criteria: appropriately timed DXA scans at ~1 year pre-menarche [predictor] and ~5 years post-menarche [dependent variable]). Regression analysis evaluated total adolescent inter-scan PA and PA over 3 maturity sub-phases as predictors of young adult bone outcomes: 1) bone mineral content (BMC), geometry and strength indices at non-dominant distal radius and femoral neck; 2) sub-head BMC; 3) lumbar spine BMC. Analyses accounted for baseline gynecological age (years pre- or post-menarche), baseline bone status, adult body size and inter-scan body size change. Gymnastics training was evaluated as a potentially independent predictor, but did not improve models for any outcomes (p<0.07). Pre-menarcheal bone traits were strong predictors of most adult outcomes (semi-partial r2 = 0.21-0.59, p≤0.001). Adult 1/3 radius and sub-head BMC were predicted by both total PA and PA 1-3 years post-menarche (p<0.03). PA 3-5 years post-menarche predicted femoral narrow neck width, endosteal diameter and buckling ratio (p<0.05). Thus, participation in organized physical activity programs throughout middle and high school may reduce lifetime fracture risk in females. PMID:25386845

  8. The Influence of Organized Physical Activity (Including Gymnastics) on Young Adult Skeletal Traits: Is Maturity Phase Important?

    PubMed

    Bernardoni, Brittney; Scerpella, Tamara A; Rosenbaum, Paula F; Kanaley, Jill A; Raab, Lindsay N; Li, Quefeng; Wang, Sijian; Dowthwaite, Jodi N

    2015-05-01

    We prospectively evaluated adolescent organized physical activity (PA) as a factor in adult female bone traits. Annual DXA scans accompanied semiannual records of anthropometry, maturity, and PA for 42 participants in this preliminary analysis (criteria: appropriately timed DXA scans at ~1 year premenarche [predictor] and ~5 years postmenarche [dependent variable]). Regression analysis evaluated total adolescent interscan PA and PA over 3 maturity subphases as predictors of young adult bone outcomes: 1) bone mineral content (BMC), geometry, and strength indices at nondominant distal radius and femoral neck; 2) subhead BMC; 3) lumbar spine BMC. Analyses accounted for baseline gynecological age (years pre- or postmenarche), baseline bone status, adult body size and interscan body size change. Gymnastics training was evaluated as a potentially independent predictor, but did not improve models for any outcomes (p > .07). Premenarcheal bone traits were strong predictors of most adult outcomes (semipartial r2 = .21-0.59, p ≤ .001). Adult 1/3 radius and subhead BMC were predicted by both total PA and PA 1-3 years postmenarche (p < .03). PA 3-5 years postmenarche predicted femoral narrow neck width, endosteal diameter, and buckling ratio (p < .05). Thus, participation in organized physical activity programs throughout middle and high school may reduce lifetime fracture risk in females. PMID:25386845

  9. Aromatic plants play an important role in promoting soil biological activity related to nitrogen cycling in an orchard ecosystem.

    PubMed

    Chen, Xinxin; Song, Beizhou; Yao, Yuncong; Wu, Hongying; Hu, Jinghui; Zhao, Lingling

    2014-02-15

    Aromatic plants can substantially improve the diversity and structure of arthropod communities, as well as reduce the number of herbivore pests and regulate the abundance of predators and parasitoids. However, it is not clear whether aromatic plants are also effective in improving soil quality by enhancing nutrient cycling. Here, field experiments are described involving intercropping with aromatic plants to investigate their effect on soil nitrogen (N) cycling in an orchard ecosystem. The results indicate that the soil organic nitrogen and available nitrogen contents increased significantly in soils intercropped with aromatic plants. Similarly, the activities of soil protease and urease increased, together with total microbial biomass involved in N cycling, including nitrifying bacteria, denitrifying bacteria and azotobacters, as well as the total numbers of bacteria and fungi. This suggests that aromatic plants improve soil N cycling and nutrient levels by enriching the soil in organic matter through the regulation of both the abundance and community structure of microorganisms, together with associated soil enzyme activity, in orchard ecosystems. PMID:24342101

  10. Sports Facilities, Shopping Centers or Homes: What Locations are Important for Adults’ Physical Activity? A Cross-Sectional Study

    PubMed Central

    Jansen, Marijke; Ettema, Dick; Pierik, Frank; Dijst, Martin

    2016-01-01

    Physical activity (PA) is influenced by the built environment. However, little is known about the types of built environment where adults spend their time, and at what levels of PA they engage in those environments. Understanding the effect of the built environment on PA requires insight into PA behavior at different types of locations (e.g., home, work, shopping centers, and sports facilities). Therefore, this study describes where adults aged 45–65 years were active with moderate-to-vigorous intensity (MVPA), and examines associations of socio-demographic factors and neighborhood with MVPA at these locations. Participants’ (N = 308) PA was measured for seven days using accelerometers and GPS-devices. Adults spent most minutes of MVPA at home and work. Highest MVPA-ratios of total time spent at a location were achieved in sports facilities and during transport. Neighborhood characteristics and socio-demographic factors such as work status, health status and household structure, had significant effects on MVPA at various locations and on total MVPA. Understanding PA behavior at various locations may provide insights that allow professionals in different domains (e.g., health, landscaping, urban planning) to develop strategies to stimulate PA. PMID:26959041

  11. Aromatic plants play an important role in promoting soil biological activity related to nitrogen cycling in an orchard ecosystem.

    PubMed

    Chen, Xinxin; Song, Beizhou; Yao, Yuncong; Wu, Hongying; Hu, Jinghui; Zhao, Lingling

    2014-02-15

    Aromatic plants can substantially improve the diversity and structure of arthropod communities, as well as reduce the number of herbivore pests and regulate the abundance of predators and parasitoids. However, it is not clear whether aromatic plants are also effective in improving soil quality by enhancing nutrient cycling. Here, field experiments are described involving intercropping with aromatic plants to investigate their effect on soil nitrogen (N) cycling in an orchard ecosystem. The results indicate that the soil organic nitrogen and available nitrogen contents increased significantly in soils intercropped with aromatic plants. Similarly, the activities of soil protease and urease increased, together with total microbial biomass involved in N cycling, including nitrifying bacteria, denitrifying bacteria and azotobacters, as well as the total numbers of bacteria and fungi. This suggests that aromatic plants improve soil N cycling and nutrient levels by enriching the soil in organic matter through the regulation of both the abundance and community structure of microorganisms, together with associated soil enzyme activity, in orchard ecosystems.

  12. Solvent-vapor concentration imparts selectivity on the propagation front during polymorphic transformation in molecular-semiconductor thin films

    NASA Astrophysics Data System (ADS)

    Purdum, Geoffrey; Gessner, Thomas; Weitz, R. Thomas; Loo, Yueh-Lin

    Post-deposition processing allows precise control over the structural development of molecular-semiconductor thin films. In particular, solvent-vapor annealing converts thin films of a core-chlorinated naphthalene diimide from its triclinic polymorph to its monoclinic polymorph. By tuning the concentration of solvent vapor, we can simultaneously impact the morphology of the resulting monoclinic thin film. At low solvent-vapor concentrations, transformation in-plane is isotropic; we observe comparable transformation rates along the b- and c-axes, resulting in plate-like domains. At high solvent-vapor concentrations, transformation along the c-axis is instead favored, resulting in the formation of needle-like domains. Extended solvent-vapor annealing at these conditions can lead to isolated needles in the active channels of field-effect transistors; these devices exhibit electron mobilities exceeding 1 cm2/Vs.

  13. Application of the Collision-Imparted Velocity Method for Analyzing the Responses of Containment and Deflector Structures to Engine Rotor Fragment Impact

    NASA Technical Reports Server (NTRS)

    Collins, T. P.; Witmer, E. A.

    1973-01-01

    An approximate analysis, termed the Collision Imparted Velocity Method (CIVM), was employed for predicting the transient structural responses of containment rings or deflector rings which are subjected to impact from turbojet-engine rotor burst fragments. These 2-d structural rings may be initially circular or arbitrarily curved and may have either uniform or variable thickness; elastic, strain hardening, and strain rate material properties are accommodated. This approximate analysis utilizes kinetic energy and momentum conservation relations in order to predict the after-impact velocities of the fragment and the impacted ring segment. This information is then used in conjunction with a finite element structural response computation code to predict the transient, large deflection responses of the ring. Similarly, the equations of motion for each fragment are solved in small steps in time. Also, some comparisons of predictions with experimental data for fragment-impacted free containment rings are presented.

  14. Trypanosoma cruzi Infection Imparts a Regulatory Program in Dendritic Cells and T Cells via Galectin-1-Dependent Mechanisms.

    PubMed

    Poncini, Carolina V; Ilarregui, Juan M; Batalla, Estela I; Engels, Steef; Cerliani, Juan P; Cucher, Marcela A; van Kooyk, Yvette; González-Cappa, Stella M; Rabinovich, Gabriel A

    2015-10-01

    Galectin-1 (Gal-1), an endogenous glycan-binding protein, is widely distributed at sites of inflammation and microbial invasion. Despite considerable progress regarding the immunoregulatory activity of this lectin, the role of endogenous Gal-1 during acute parasite infections is uncertain. In this study, we show that Gal-1 functions as a negative regulator to limit host-protective immunity following intradermal infection with Trypanosoma cruzi. Concomitant with the upregulation of immune inhibitory mediators, including IL-10, TGF-β1, IDO, and programmed death ligand 2, T. cruzi infection induced an early increase of Gal-1 expression in vivo. Compared to their wild-type (WT) counterpart, Gal-1-deficient (Lgals1(-/-)) mice exhibited reduced mortality and lower parasite load in muscle tissue. Resistance of Lgals1(-/-) mice to T. cruzi infection was associated with a failure in the activation of Gal-1-driven tolerogenic circuits, otherwise orchestrated by WT dendritic cells, leading to secondary dysfunction in the induction of CD4(+)CD25(+)Foxp3(+) regulatory T cells. This effect was accompanied by an increased number of CD8(+) T cells and higher frequency of IFN-γ-producing CD4(+) T cells in muscle tissues and draining lymph nodes as well as reduced parasite burden in heart and hindlimb skeletal muscle. Moreover, dendritic cells lacking Gal-1 interrupted the Gal-1-mediated tolerogenic circuit and reinforced T cell-dependent anti-parasite immunity when adoptively transferred into WT mice. Thus, endogenous Gal-1 may influence T. cruzi infection by fueling tolerogenic circuits that hinder anti-parasite immunity.

  15. Predictors of Unprotected Sex among Young Sexually Active African American, Hispanic, and White MSM: The Importance of Ethnicity and Culture

    PubMed Central

    Fernández, M. Isabel; Harper, Gary W.; Hidalgo, Marco A.; Jamil, Omar B.; Torres, Rodrigo Sebastián

    2010-01-01

    Despite the recognized need for culturally tailored HIV prevention interventions for gay, bisexual, and questioning youth, few studies have examined if predictors of unprotected sex vary for youth from different ethnic groups. This study reports on a sample of 189 gay, bisexual, and questioning youth (age 15–22) from three racial/ethnic backgrounds (African American, Hispanic, and White) recruited in Chicago, IL and Miami-Dade and Broward Counties, Florida. For African American youth, being in a long-term relationship, having been kicked out of the home for having sex with men, and younger age at initiation of sexual behavior were associated with unprotected sex. For Hispanic youth, higher ethnic identification and older age at initiation of sexual behavior were associated with unprotected sex. For White youth, no predictors were associated with unprotected sex. Our findings point to the importance of understanding the varying predictors of unprotected sex and integrating them into tailored prevention interventions. PMID:17721725

  16. A trustful relationship—the importance for relatives to actively participate in the meeting with the physician

    PubMed Central

    2013-01-01

    In previous research, no uniform picture emerged of the role of relatives in the meeting between an elderly patient and a physician. Knowledge about relatives’ experiences of the meeting between an elderly patient and a physician will help healthcare practitioners better understand the role of relatives during the meeting and how practitioners can assist relatives in assuming their supporting role more efficiently. The purpose of this study is to explore experiences of relatives of meeting with the physician in a hospital setting when an elderly patient is discharged from hospital care to home care, in order to identify aspects that may facilitate relatives in taking up their role in a more efficient manner. This descriptive and exploratory study is based on 20 interviews with relatives. The result shows that the physician's communication style influences the meeting between the relative, the elderly patient, and the physician, and that this style is the result of power and interaction. A trustful relationship during the meeting between the relative and the physician can increase the relative's feeling of confidence with the healthcare organization and treatment of the elderly patient. The relative has an important supporting role in the care for the elderly family member, both in the hospital and the home setting. It is likely that the relative's value as a resource, for both the patient and the physician, increases as the relative experiences feelings of confidence in the meeting with the physician. It is therefore of value to increase our knowledge about the conditions and circumstances facilitating and/or hampering the meeting between the relative and the physician. The result stresses the importance of encouraging relatives to participate in the meeting. Physicians need more guidance and training in communication skills, respectful demeanor, and collaboration while meeting the relatives. PMID:23694680

  17. Evidence that Highly Conserved Residues of Transmembrane Segment 6 of Escherichia coli MntH Are Important for Transport Activity

    PubMed Central

    Haemig, Heather A.H.; Moen, Patrick J.; Brooker, Robert J.

    2010-01-01

    Nramp (natural resistance-associated macrophage protein) family members have been characterized in mammals, yeast, and bacteria as divalent metal ion/H+ symporters. In previous work, a bioinformatic approach was used for the identification of residues that are conserved within the Nramp family [ref.1. Haemig, H.A. and R.J. Brooker, (2004) J Membr. Biol, 201(2): 97-107]. Based on site-directed mutagenesis of highly conserved negatively charged residues, a model was proposed for the metal binding site of the E.coli homolog, MntH. In this study, we have focused on the highly conserved residues, including two histidines, of transmembrane segment 6 (TMS-6). Multiple mutants were made at the eight conserved sites (i.e., Gly-205, Ala-206, Met-209, Pro-210, His-211, Leu-215, His-216, and Ser-217) in TMS-6 of E. coli MntH. Double mutants involving His-211 and His-216 were also made. The results indicate the side chain volume of these residues is critically important for function. In most cases, only substitutions that are closest in side chain volume still permit transport. In addition, the Km for metal binding is largely unaffected by mutations in TMS-6, whereas Vmax values were decreased in all mutants characterized kinetically. Thus, these residues do not appear to play a role in metal binding. Instead, they may comprise an important face on TMS-6 that is critical for protein conformational changes during transport. Also, in contrast to other studies, our data do not strongly indicate that the conserved histidine residues play a role in pH regulation of metal transport. PMID:20441230

  18. Importance of aerosol composition and mixing state for cloud droplet activation over the Arctic pack ice in summer

    NASA Astrophysics Data System (ADS)

    Leck, C.; Svensson, E.

    2015-03-01

    Concentrations of cloud condensation nuclei (CCN) were measured throughout an expedition by icebreaker around the central Arctic Ocean, including a 3 week ice drift operation at 87° N, from 3 August to 9 September 2008. In agreement with previous observations in the area and season, median daily CCN concentrations at 0.2% water vapour supersaturation (SS) were typically in the range of 15 to 30 cm-3, but concentrations varied by 2 to 3 orders of magnitude over the expedition and were occasionally below 1 cm-3. The CCN concentrations were highest near the ice edge and fell by a factor of 3 in the first 48 h of transport from the open sea into the pack ice region. For longer transport times they increased again, indicating a local source over the pack ice, suggested to be polymer gels, via drops injected into the air by bubbles bursting on open leads. We inferred the properties of the unexplained non-water soluble aerosol fraction that was necessary for reproducing the observed concentrations of CCN. This was made possible by assuming Köhler theory and simulating the cloud nucleation process using a Lagrangian adiabatic air parcel model that solves the kinetic formulation for condensation of water on size resolved aerosol particles. We propose that the portion of the internally/externally mixed water insoluble particles was larger in the corresponding smaller aerosol size ranges. These particles were physically and chemically behaving as polymer gels: the interaction of the hydrophilic and hydrophobic entities on the structures of polymer gels during cloud droplet activation would at first only show a partial wetting character and only weak hygroscopic growth. Given time, a high CCN activation efficiency is achieved, which is promoted by the hydrophilicity or surface-active properties of the gels. Thus the result in this study argues that the behaviour of the high Arctic aerosol in CCN-counters operating at water vapour SSs > 0.4% (high relative humidities) may not

  19. A stress-responsive late embryogenesis abundant protein 7 (CsLEA7) of tea [Camellia sinensis (L.) O. Kuntze] encodes for a chaperone that imparts tolerance to Escherichia coli against stresses.

    PubMed

    Paul, Asosii; Singh, Sewa; Sharma, Shweta; Kumar, Sanjay

    2014-11-01

    The present study characterized CsLEA7, a group 7 late embryogenesis abundant (LEA) gene, from tea [Camellia sinensis (L.) O. Kuntze]. The gene had an open reading frame of 462 base pairs encoding 153 amino acids with calculated molecular weight of 16.63 kDa and an isoelectric point (pI) of 4.93. Analysis revealed CsLEA7 to be an intrinsically ordered protein consisting of nine β-strands and two α-helices. CsLEA7 expressed ubiquitously in all the tissues analyzed with highest level of transcripts in mature leaf as compared to in flower bud, younger leaves, stem and fruit. Expression was the least in root tissue. CsLEA7 exhibited up-regulation in response to low temperature, polyethylene glycol-8000, sodium chloride and hydrogen peroxide in tea. Analysis of the promoter of CsLEA7 revealed a core promoter element and distinct cis-acting regulatory elements regulating gene expression under abiotic stresses. CsLEA7 exhibited chaperonic activity as evinced by protection to malate dehydrogenase against heat denaturation assay. Recombinant Escherichia coli cells producing CsLEA7 exhibited improved tolerance against diverse cues: polyethylene glycol-8000, sodium chloride, hydrogen peroxide and low temperature signifying its role in imparting stress tolerance.

  20. A stress-responsive late embryogenesis abundant protein 7 (CsLEA7) of tea [Camellia sinensis (L.) O. Kuntze] encodes for a chaperone that imparts tolerance to Escherichia coli against stresses.

    PubMed

    Paul, Asosii; Singh, Sewa; Sharma, Shweta; Kumar, Sanjay

    2014-11-01

    The present study characterized CsLEA7, a group 7 late embryogenesis abundant (LEA) gene, from tea [Camellia sinensis (L.) O. Kuntze]. The gene had an open reading frame of 462 base pairs encoding 153 amino acids with calculated molecular weight of 16.63 kDa and an isoelectric point (pI) of 4.93. Analysis revealed CsLEA7 to be an intrinsically ordered protein consisting of nine β-strands and two α-helices. CsLEA7 expressed ubiquitously in all the tissues analyzed with highest level of transcripts in mature leaf as compared to in flower bud, younger leaves, stem and fruit. Expression was the least in root tissue. CsLEA7 exhibited up-regulation in response to low temperature, polyethylene glycol-8000, sodium chloride and hydrogen peroxide in tea. Analysis of the promoter of CsLEA7 revealed a core promoter element and distinct cis-acting regulatory elements regulating gene expression under abiotic stresses. CsLEA7 exhibited chaperonic activity as evinced by protection to malate dehydrogenase against heat denaturation assay. Recombinant Escherichia coli cells producing CsLEA7 exhibited improved tolerance against diverse cues: polyethylene glycol-8000, sodium chloride, hydrogen peroxide and low temperature signifying its role in imparting stress tolerance. PMID:25052187

  1. Increase of electrodermal activity of heart meridian during physical exercise: the significance of electrical values in acupuncture and diagnostic importance.

    PubMed

    Pontarollo, Francesco; Rapacioli, Giuliana; Bellavite, Paolo

    2010-08-01

    Electric field measurements of skin potential and electrical currents are physiological indicators of electrodermal activity (EDA) and have been associated with a variety of sensory, cognitive and emotional stimuli. The aim of this study was to investigate the EDA at some hand acupoints before, during and after a physical exercise. EDA of eight points located at the corner of fingernails of hands was measured in 10 healthy young volunteers before, during and after a 14-min acute exercise in a bicycle ergometer. In pre-exercise resting state the parameters were stable and similar between the 8 different tested points, while during exercise a significant increase of current (from 1000-2000 to 4000-8000 nA) was observed, with the maximal values related to the point located on the ulnar side of the little finger, at the base of the nail, corresponding to the Shao chong (HT9) of heart meridian.

  2. Increasing the catalytic activity of Bilirubin oxidase from Bacillus pumilus: Importance of host strain and chaperones proteins.

    PubMed

    Gounel, Sébastien; Rouhana, Jad; Stines-Chaumeil, Claire; Cadet, Marine; Mano, Nicolas

    2016-07-20

    Aggregation of recombinant proteins into inclusion bodies (IBs) is the main problem of the expression of multicopper oxidase in Escherichia coli. It is usually attributed to inefficient folding of proteins due to the lack of copper and/or unavailability of chaperone proteins. The general strategies reported to overcome this issue have been focused on increasing the intracellular copper concentration. Here we report a complementary method to optimize the expression in E. coli of a promising Bilirubin oxidase (BOD) isolated from Bacillus pumilus. First, as this BOD has a disulfide bridge, we switched E.coli strain from BL21 (DE3) to Origami B (DE3), known to promote the formation of disulfide bridges in the bacterial cytoplasm. In a second step, we investigate the effect of co-expression of chaperone proteins on the protein production and specific activity. Our strategy allowed increasing the final amount of enzyme by 858% and its catalytic rate constant by 83%. PMID:27165502

  3. DNA polymerase κ-dependent DNA synthesis at stalled replication forks is important for CHK1 activation

    PubMed Central

    Bétous, Rémy; Pillaire, Marie-Jeanne; Pierini, Laura; van der Laan, Siem; Recolin, Bénédicte; Ohl-Séguy, Emma; Guo, Caixia; Niimi, Naoko; Grúz, Petr; Nohmi, Takehiko; Friedberg, Errol; Cazaux, Christophe; Maiorano, Domenico; Hoffmann, Jean-Sébastien

    2013-01-01

    Formation of primed single-stranded DNA at stalled replication forks triggers activation of the replication checkpoint signalling cascade resulting in the ATR-mediated phosphorylation of the Chk1 protein kinase, thus preventing genomic instability. By using siRNA-mediated depletion in human cells and immunodepletion and reconstitution experiments in Xenopus egg extracts, we report that the Y-family translesion (TLS) DNA polymerase kappa (Pol κ) contributes to the replication checkpoint response and is required for recovery after replication stress. We found that Pol κ is implicated in the synthesis of short DNA intermediates at stalled forks, facilitating the recruitment of the 9-1-1 checkpoint clamp. Furthermore, we show that Pol κ interacts with the Rad9 subunit of the 9-1-1 complex. Finally, we show that this novel checkpoint function of Pol κ is required for the maintenance of genomic stability and cell proliferation in unstressed human cells. PMID:23799366

  4. Increase of electrodermal activity of heart meridian during physical exercise: the significance of electrical values in acupuncture and diagnostic importance.

    PubMed

    Pontarollo, Francesco; Rapacioli, Giuliana; Bellavite, Paolo

    2010-08-01

    Electric field measurements of skin potential and electrical currents are physiological indicators of electrodermal activity (EDA) and have been associated with a variety of sensory, cognitive and emotional stimuli. The aim of this study was to investigate the EDA at some hand acupoints before, during and after a physical exercise. EDA of eight points located at the corner of fingernails of hands was measured in 10 healthy young volunteers before, during and after a 14-min acute exercise in a bicycle ergometer. In pre-exercise resting state the parameters were stable and similar between the 8 different tested points, while during exercise a significant increase of current (from 1000-2000 to 4000-8000 nA) was observed, with the maximal values related to the point located on the ulnar side of the little finger, at the base of the nail, corresponding to the Shao chong (HT9) of heart meridian. PMID:20621275

  5. Prediction of prognosis of ALS: Importance of active denervation findings of the cervical-upper limb area and trunk area.

    PubMed

    Sato, Yoko; Nakatani, Eiji; Watanabe, Yasuhiro; Fukushima, Masanori; Nakashima, Kenji; Kannagi, Mari; Kanatani, Yasuhiro; Mizushima, Hiroshi

    2015-11-01

    Amyotrophic lateral sclerosis (ALS) is a motor neuron disease characterized by serious muscle atrophy and weakness. The purpose of this study was to find prognostic factors in patients with mild ALS using application forms for the Specified Disease Treatment Research Program in Japan. We classified ALS as mild, moderate and severe. The subjects consisted of 363 patients with mild ALS who underwent needle electromyography at registration and were followed for more than one year. Time to progression to severe ALS and time to deterioration of activities of daily living such as speech dysfunction, upper limb dysfunction, and walking disability were used as outcomes. Cox proportional hazards model analysis was performed to identify prognostic factors. Of the patients with initially mild ALS, 38.3% (139/363) had progressed severe ALS at the last follow-up. In multivariate analysis of time to progression to severe ALS, bulbar onset (hazard ratio [95% confidence interval]: 1.68 [1.13-2.49], p = 0.010), tongue atrophy (1.69 [1.14-2.51], p = 0.009), dyspnea (1.57 [1.02-2.41], p = 0.042) and active denervation findings (ADFs) of the cervical-upper limb area (1.81 [1.25-2.63], p = 0.002) emerged as prognostic factors. Furthermore ADFs in the trunk area were prognostic factors for upper limb dysfunction and walking disability (1.72 [1.05-2.81], p = 0.031, and 1.97 [1.09-3.59], p = 0.026). In conclusion ADFs of the cervical-upper limb area and trunk area were prognostic factors in ALS patients.

  6. Self-assembly with orthogonal-imposed stimuli to impart structure and confer magnetic function to electrodeposited hydrogels.

    PubMed

    Li, Ying; Liu, Yi; Gao, Tieren; Zhang, Boce; Song, Yingying; Terrell, Jessica L; Barber, Nathan; Bentley, William E; Takeuchi, Ichiro; Payne, Gregory F; Wang, Qin

    2015-05-20

    A magnetic nanocomposite film with the capability of reversibly collecting functionalized magnetic particles was fabricated by simultaneously imposing two orthogonal stimuli (electrical and magnetic). We demonstrate that cathodic codeposition of chitosan and Fe3O4 nanoparticles while simultaneously applying a magnetic field during codeposition can (i) organize structure, (ii) confer magnetic properties, and (iii) yield magnetic films that can perform reversible collection/assembly functions. The magnetic field triggered the self-assembly of Fe3O4 nanoparticles into hierarchical "chains" and "fibers" in the chitosan film. For controlled magnetic properties, the Fe3O4-chitosan film was electrodeposited in the presence of various strength magnetic fields and different deposition times. The magnetic properties of the resulting films should enable broad applications in complex devices. As a proof of concept, we demonstrate the reversible capture and release of green fluorescent protein (EGFP)-conjugated magnetic microparticles by the magnetic chitosan film. Moreover, antibody-functionalized magnetic microparticles were applied to capture cells from a sample, and these cells were collected, analyzed, and released by the magnetic chitosan film, paving the way for applications such as reusable biosensor interfaces (e.g., for pathogen detection). To our knowledge, this is the first report to apply a magnetic field during the electrodeposition of a hydrogel to generate magnetic soft matter. Importantly, the simple, rapid, and reagentless fabrication methodologies demonstrated here are valuable features for creating a magnetic device interface. PMID:25923335

  7. Molecular and Functional Characterization of a Wheat B2 Protein Imparting Adverse Temperature Tolerance and Influencing Plant Growth.

    PubMed

    Singh, Akanksha; Khurana, Paramjit

    2016-01-01

    Genomic attempts were undertaken to elucidate the plant developmental responses to heat stress, and to characterize the roles of B2 protein in mediating those responses. A wheat expressed sequence tag for B2 protein was identified which was cloned and characterized to assess its functional relevance causing plant growth and development during stress adaptation. Here, we show that wheat B2 protein is highly expressed in root and shoot tissues as well as in developing seed tissues under high temperature stress conditions. Morphological studies of transgenic Arabidopsis overexpressing gene encoding wheat B2 protein and Δb2 mutant plants were studied at major developmental stages. The stunted growth phenotype of mutant plants, together with hypocotyl and root elongation analysis of transgenic plants showed that B2 protein exhibits a crucial role in plant growth and development. Additional physiological analyses highlights the role of B2 protein in increased tolerance to heat and cold stresses by maintaining high chlorophyll content, strong activity of photosystem II and less membrane damage of overexpression transgenics as compared with the wild-type. Furthermore, the constitutive overexpression of TaB2 in Arabidopsis resulted in ABA hypersensitivity. Taken together, these studies suggest a novel perspectives of B2 protein in plant development and in mediating the thermal stress tolerance.

  8. Molecular and Functional Characterization of a Wheat B2 Protein Imparting Adverse Temperature Tolerance and Influencing Plant Growth

    PubMed Central

    Singh, Akanksha; Khurana, Paramjit

    2016-01-01

    Genomic attempts were undertaken to elucidate the plant developmental responses to heat stress, and to characterize the roles of B2 protein in mediating those responses. A wheat expressed sequence tag for B2 protein was identified which was cloned and characterized to assess its functional relevance causing plant growth and development during stress adaptation. Here, we show that wheat B2 protein is highly expressed in root and shoot tissues as well as in developing seed tissues under high temperature stress conditions. Morphological studies of transgenic Arabidopsis overexpressing gene encoding wheat B2 protein and Δb2 mutant plants were studied at major developmental stages. The stunted growth phenotype of mutant plants, together with hypocotyl and root elongation analysis of transgenic plants showed that B2 protein exhibits a crucial role in plant growth and development. Additional physiological analyses highlights the role of B2 protein in increased tolerance to heat and cold stresses by maintaining high chlorophyll content, strong activity of photosystem II and less membrane damage of overexpression transgenics as compared with the wild-type. Furthermore, the constitutive overexpression of TaB2 in Arabidopsis resulted in ABA hypersensitivity. Taken together, these studies suggest a novel perspectives of B2 protein in plant development and in mediating the thermal stress tolerance. PMID:27242843

  9. Molecular and Functional Characterization of a Wheat B2 Protein Imparting Adverse Temperature Tolerance and Influencing Plant Growth.

    PubMed

    Singh, Akanksha; Khurana, Paramjit

    2016-01-01

    Genomic attempts were undertaken to elucidate the plant developmental responses to heat stress, and to characterize the roles of B2 protein in mediating those responses. A wheat expressed sequence tag for B2 protein was identified which was cloned and characterized to assess its functional relevance causing plant growth and development during stress adaptation. Here, we show that wheat B2 protein is highly expressed in root and shoot tissues as well as in developing seed tissues under high temperature stress conditions. Morphological studies of transgenic Arabidopsis overexpressing gene encoding wheat B2 protein and Δb2 mutant plants were studied at major developmental stages. The stunted growth phenotype of mutant plants, together with hypocotyl and root elongation analysis of transgenic plants showed that B2 protein exhibits a crucial role in plant growth and development. Additional physiological analyses highlights the role of B2 protein in increased tolerance to heat and cold stresses by maintaining high chlorophyll content, strong activity of photosystem II and less membrane damage of overexpression transgenics as compared with the wild-type. Furthermore, the constitutive overexpression of TaB2 in Arabidopsis resulted in ABA hypersensitivity. Taken together, these studies suggest a novel perspectives of B2 protein in plant development and in mediating the thermal stress tolerance. PMID:27242843

  10. Ixodes (Ixodes) scapularis (Acari:Ixodidae): redescription of all active stages, distribution, hosts, geographical variation, and medical and veterinary importance.

    PubMed

    Keirans, J E; Hutcheson, H J; Durden, L A; Klompen, J S

    1996-05-01

    The blacklegged tick, Ixodes (Ixodes) scapularis Say, 1821, is redescribed, based on laboratory reared specimens originating in Bulloch County, Georgia. Information on distribution, host associations, morphological variation, and medical/veterinary importance is also presented. A great deal of recent work has focused on this species because it is the principal vector of the agent of Lyme disease (Borrelia burgdorferi Johnson, Schmidt, Hyde, Steigerwaldt & Brenner) in eastern North America. Its distribution appears to be expanding, and includes the state of Florida in the southeastern United States north to the provinces of Nova Scotia and Prince Edward Island, Canada, west to North and South Dakota, United States, and south to the state of Coahuila, Mexico. Although I. scapularis feeds on at least 125 species of North American vertebrates (54 mammalian, 57 avian, and 14 lizard species), analysis of the U.S. National Tick Collection holdings show that white-tailed deer, Odocoileus virginianus (Zimmermann), cattle, Bos taurus L., dogs, Canis lupus L., and other medium-to-large sized mammals are important hosts for adults as are native mice and other small mammals, certain ground-frequenting birds, skinks, and glass lizards for nymphs and larvae. This tick is a polytypic species exhibiting north-south and east-west morphological clines. Analysis of variance and Student-Newman-Keuls multiple comparisons revealed significant interpopulational variation that is expressed most significantly in the nymphal stage. Nymphs from northern (Minnesota, Massachusetts, Maryland) populations had relatively larger basis capituli with shorter cornua (except Maryland) than southern (North Carolina, Georgia) populations. Midwestern populations (Minnesota, Missouri) differed from eastern populations (Massachusetts, Maryland, North Carolina, Georgia) in idiosomal characters (broader scuta, larger coxae III, and IV). In addition to Lyme disease, this tick is also a primary vector of the

  11. Identification of functionally important residues in the silkmoth pheromone biosynthesis-activating neuropeptide receptor, an insect ortholog of the vertebrate Neuromedin U Receptor

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The biosynthesis of sex pheromone components in many lepidopteran insects is regulated by interactions between pheromone biosynthesis-activating neuropeptide (PBAN) and the PBAN receptor (PBANR), a class-A G-protein-coupled receptor (GPCR). To identify functionally important amino acid residues in t...

  12. Fatty Amines from Little Black Ants, Monomorium minimum, and Their Biological Activities Against Red Imported Fire Ants, Solenopsis invicta.

    PubMed

    Wang, Lei; Chen, Jian

    2015-08-01

    Red imported fire ants, Solenopsis invicta, are significant invasive pests. Certain native ant species can compete with S. invicta, such as the little black ant, Monomorium minimum. Defensive secretions may contribute to the competition capacity of native ants. The chemistry of ant defensive secretions in the genus Monomorium has been subjected to extensive research. The insecticidal alkaloids, 2,5-dialkyl-pyrrolidines and 2,5-dialkyl-pyrrolines have been reported to dominate the venom of M. minimum. In this study, analysis of defensive secretions of workers and queens of M. minimum revealed two primary amines, decylamine and dodecylamine. Neither amine has been reported previously from natural sources. Toxicity and digging suppression by these two amines against S. invicta were examined. Decylamine had higher toxicity to S. invicta workers than dodecylamine, a quicker knockdown effect, and suppressed the digging behavior of S. invicta workers at lower concentration. However, the amount of fatty amines in an individual ant was not enough to knockdown a fire ant or suppress its digging behavior. These amines most likely work in concert with other components in the chemical defense of M. minimum.

  13. Halopriming of seeds imparts tolerance to NaCl and PEG induced stress in Vigna radiata (L.) Wilczek varieties.

    PubMed

    Jisha, K C; Puthur, Jos T

    2014-07-01

    The investigation was carried out to study the effect of halopriming on NaCl and polyethylene glycol-6000 (PEG-6000) induced stress tolerance potential of three Vigna radiata (L.) Wilczek varieties, with varied abiotic stress tolerance potential. Halopriming is a seed priming technique in which the seeds were soaked in various salt solutions (in this study NaCl was used). The results of the study indicated that the application of stresses (both NaCl and PEG) induced retardation of growth attributes (measured in terms of shoot length, fresh weight, dry weight) and decrease in physiological attributes like total chlorophyll content, metabolites, photosynthetic and mitochondrial activity of the seedlings in all three V. radiata (L.) varieties. However, halopriming of the seeds could reduce the extent of decrease in these biological attributes. NaCl and PEG stress also caused increase in MDA content (a product of membrane lipid peroxidation) in all the varieties studied and this increase was significantly minimized under halopriming. From the present investigation it was evident that among the green gram varieties studied, Pusa Vishal, a NaCl tolerant variety showed enhanced tolerance to NaCl and PEG induced stress, when the seeds were subjected to halopriming followed by Pusa Ratna (stress sensitive variety). Pusa 9531 (drought tolerant variety) also showed positive halopriming effects but it was less significant when compared to other two varieties. It could be concluded that halopriming improved the drought and salinity stress tolerance potential of all varieties and it was significantly higher in the Pusa Vishal as compared to Pusa 9531 and Pusa Ratna.

  14. Physical activity and cardiorespiratory fitness as major markers of cardiovascular risk: their independent and interwoven importance to health status.

    PubMed

    Myers, Jonathan; McAuley, Paul; Lavie, Carl J; Despres, Jean-Pierre; Arena, Ross; Kokkinos, Peter

    2015-01-01

    The evolution from hunting and gathering to agriculture, followed by industrialization, has had a profound effect on human physical activity (PA) patterns. Current PA patterns are undoubtedly the lowest they have been in human history, with particularly marked declines in recent generations, and future projections indicate further declines around the globe. Non-communicable health problems that afflict current societies are fundamentally attributable to the fact that PA patterns are markedly different than those for which humans were genetically adapted. The advent of modern statistics and epidemiological methods has made it possible to quantify the independent effects of cardiorespiratory fitness (CRF) and PA on health outcomes. Based on more than five decades of epidemiological studies, it is now widely accepted that higher PA patterns and levels of CRF are associated with better health outcomes. This review will discuss the evidence supporting the premise that PA and CRF are independent risk factors for cardiovascular disease (CVD) as well as the interplay between both PA and CRF and other CVD risk factors. A particular focus will be given to the interplay between CRF, metabolic risk and obesity.

  15. Site-directed mutagenesis of the human DNA repair enzyme HAP1: identification of residues important for AP endonuclease and RNase H activity.

    PubMed

    Barzilay, G; Walker, L J; Robson, C N; Hickson, I D

    1995-05-11

    HAP1 protein, the major apurinic/apyrimidinic (AP) endonuclease in human cells, is a member of a homologous family of multifunctional DNA repair enzymes including the Escherichia coli exonuclease III and Drosophila Rrp1 proteins. The most extensively characterised member of this family, exonuclease III, exhibits both DNA- and RNA-specific nuclease activities. Here, we show that the RNase H activity characteristic of exonuclease III has been conserved in the human homologue, although the products resulting from RNA cleavage are dissimilar. To identify residues important for enzymatic activity, five mutant HAP1 proteins containing single amino acid substitutions were purified and analysed in vitro. The substitutions were made at sites of conserved amino acids and targeted either acidic or histidine residues because of their known participation in the active sites of hydrolytic nucleases. One of the mutant proteins (replacement of Asp-219 by alanine) showed a markedly reduced enzymatic activity, consistent with a greatly diminished capacity to bind DNA and RNA. In contrast, replacement of Asp-90, Asp-308 or Glu-96 by alanine led to a reduction in enzymatic activity without significantly compromising nucleic acid binding. Replacement of His-255 by alanine led to only a very small reduction in enzymatic activity. Our data are consistent with the presence of a single catalytic active site for the DNA- and RNA-specific nuclease activities of the HAP1 protein. PMID:7784208

  16. Importance of biologically active aurora-like ultraviolet emission: stochastic irradiation of Earth and Mars by flares and explosions.

    PubMed

    Smith, David S; Scalo, John; Wheeler, J Craig

    2004-10-01

    Habitable planets will be subject to intense sources of ionizing radiation and fast particles from a variety of sources--from the host star to distant explosions--on a variety of timescales. Monte Carlo calculations of high-energy irradiation suggest that the surfaces of terrestrial-like planets with thick atmospheres (column densities greater than about 100 g cm(-2)) are well protected from directly incident X-rays and gamma-rays, but we find that sizeable fractions of incident ionizing radiation from astrophysical sources can be redistributed to biologically and chemically important ultraviolet wavelengths, a significant fraction of which can reach the surface. This redistribution is mediated by secondary electrons, resulting from Compton scattering and X-ray photoabsorption, the energies of which are low enough to excite and ionize atmospheric molecules and atoms, resulting in a rich aurora-like spectrum. We calculate the fraction of energy redistributed into biologically and chemically important wavelength regions for spectra characteristic of stellar flares and supernovae using a Monte-Carlo transport code and then estimate the fraction of this energy that is transmitted from the atmospheric altitudes of redistribution to the surface for a few illustrative cases. For atmospheric models corresponding to the Archean Earth, we assume no significant ultraviolet absorbers, only Rayleigh scattering, and find that the fraction of incident ionizing radiation that is received at the surface in the form of redistributed ultraviolet in the biologically relevant 200-320 nm region (UV-C and UV-B bands) can be up to 4%. On the present-day Earth with its ultraviolet ozone shield, this fraction is found to be 0.2%. Both values are many orders of magnitude higher than the fraction of direct ionizing radiation reaching the surface. This result implies that planetary organisms will be subject to mutationally significant, if intermittent, fluences of UV-B and harder radiation

  17. Slow Climate Velocities in Mountain Streams Impart Thermal Resistance to Cold-Water Refugia Across the West

    NASA Astrophysics Data System (ADS)

    Isaak, D.; Young, M.; Luce, C.; Hostetler, S.; Wenger, S. J.; Peterson, E.; Ver Hoef, J.

    2015-12-01

    Mountain streams provide important headwater refugia for native fish, amphibians, and other cold-water fauna globally. Although the well documented existence of such refugia indicates some level of resistance to ongoing environmental change, stream warming associated with climate change raises questions about their future persistence. Moreover, evidence exists that air temperatures are warming faster at higher elevations, and some stream temperature models predict that cold streams associated with snowmelt hydrologies will be most sensitive to air temperature increases (i.e. high ratio of stream Δ˚C:air Δ˚C). Here, we estimate stream sensitivities to climate forcing using long-term monitoring records from 927 sites across the topographically complex northwestern U.S. Sensitivity values are combined with high-resolution NorWeST stream temperature scenarios (website: http://www.fs.fed.us/rm/boise/AWAE/projects/NorWeST.html) to map climate velocities at 1 kilometer resolution throughout the 450,000 stream kilometers in the regional network. Our results suggest that cold mountain streams are often 'double buffered' against the thermal effects of climate change due to low sensitivities (0.3ºC/ºC) and steep gradients, which translated to very slow climate velocities (<0.35 km/decade for streams >3% slope) from 1968-2011 when air temperatures warmed at the rate of 0.2ºC/decade. Alternative scenarios based on aggressive air temperature warming rates (2x historical rates) and higher sensitivity values of cold streams suggests velocities will remain low in mountain streams due to the dominant effects of steep channel slope and strong local temperature gradients. These results reinforce earlier predictions from high-resolution species distribution models that show which watersheds are most likely to host resilient native trout populations across the West later this century (Climate Shield project website: http://www.fs

  18. Line-driven disk winds in active galactic nuclei: The critical importance of ionization and radiative transfer

    SciTech Connect

    Higginbottom, Nick; Knigge, Christian; Matthews, James H.; Proga, Daniel; Long, Knox S.; Sim, Stuart A.

    2014-07-01

    Accretion disk winds are thought to produce many of the characteristic features seen in the spectra of active galactic nuclei (AGNs) and quasi-stellar objects (QSOs). These outflows also represent a natural form of feedback between the central supermassive black hole and its host galaxy. The mechanism for driving this mass loss remains unknown, although radiation pressure mediated by spectral lines is a leading candidate. Here, we calculate the ionization state of, and emergent spectra for, the hydrodynamic simulation of a line-driven disk wind previously presented by Proga and Kallman. To achieve this, we carry out a comprehensive Monte Carlo simulation of the radiative transfer through, and energy exchange within, the predicted outflow. We find that the wind is much more ionized than originally estimated. This is in part because it is much more difficult to shield any wind regions effectively when the outflow itself is allowed to reprocess and redirect ionizing photons. As a result, the calculated spectrum that would be observed from this particular outflow solution would not contain the ultraviolet spectral lines that are observed in many AGN/QSOs. Furthermore, the wind is so highly ionized that line driving would not actually be efficient. This does not necessarily mean that line-driven winds are not viable. However, our work does illustrate that in order to arrive at a self-consistent model of line-driven disk winds in AGN/QSO, it will be critical to include a more detailed treatment of radiative transfer and ionization in the next generation of hydrodynamic simulations.

  19. In vitro and in vivo [corrected] activity of eugenol oil (Eugenia caryophylata) against four important postharvest apple pathogens.

    PubMed

    Amiri, Achour; Dugas, Robert; Pichot, Anne L; Bompeix, Gilbert

    2008-08-15

    The activity of eugenol oil was evaluated in vitro and in vivo against four apple pathogens namely Phlyctema vagabunda, Penicillium expansum, Botrytis cinerea and Monilinia fructigena. The minimum inhibitory concentration (MIC) of eugenol incorporated in malt extract agar medium was found to be 2 mg ml(-1). Mycelial growth of the four test pathogens was completely inhibited when treated with 150 microl l(-1) of volatile eugenol whether at 4 or 20 degrees C. Conidia of P. vagabunda, P. expansum, M. fructigena and B. cinerea suspended for 2 min in eugenol solution at 2 mg ml(-1) heated to 50 degrees C germinated at rates of 19, 37, 38 and 39%, respectively. Three different eugenol formulations (Tween 80, ethoxylate and lecithin) were tested for their in vivo efficacy against the tested pathogens on apples. Ethoxylate- and Tween 80-eugenol formulations applied at room temperature were ineffective in reducing disease incidence. When heated to 50 degrees C, both formulations induced phytotoxicity on apple surface and caused cuticle damages as revealed by scanning electronic microscopic observations. A mixture of eugenol at 2 mg ml(-1) and soy lecithin at 50 mg ml(-1) suppressed the phytotoxic symptoms produced by eugenol on apples and reduced the disease incidence of P. expansum, P. vagabunda, B. cinerea and M. fructigena to less than 7, 6, 4 and 2% respectively after 6 months of storage at 2 degrees C. The application of heated lecithin-formulated eugenol could become a successful alternative to the traditional fungicides used in postharvest disease management of apple fruit.

  20. FgPrp4 Kinase Is Important for Spliceosome B-Complex Activation and Splicing Efficiency in Fusarium graminearum.

    PubMed

    Gao, Xuli; Jin, Qiaojun; Jiang, Cong; Li, Yang; Li, Chaohui; Liu, Huiquan; Kang, Zhensheng; Xu, Jin-Rong

    2016-04-01

    PRP4 encodes the only kinase among the spliceosome components. Although it is an essential gene in the fission yeast and other eukaryotic organisms, the Fgprp4 mutant was viable in the wheat scab fungus Fusarium graminearum. Deletion of FgPRP4 did not block intron splicing but affected intron splicing efficiency in over 60% of the F. graminearum genes. The Fgprp4 mutant had severe growth defects and produced spontaneous suppressors that were recovered in growth rate. Suppressor mutations were identified in the PRP6, PRP31, BRR2, and PRP8 orthologs in nine suppressor strains by sequencing analysis with candidate tri-snRNP component genes. The Q86K mutation in FgMSL1 was identified by whole genome sequencing in suppressor mutant S3. Whereas two of the suppressor mutations in FgBrr2 and FgPrp8 were similar to those characterized in their orthologs in yeasts, suppressor mutations in Prp6 and Prp31 orthologs or FgMSL1 have not been reported. Interestingly, four and two suppressor mutations identified in FgPrp6 and FgPrp31, respectively, all are near the conserved Prp4-phosphorylation sites, suggesting that these mutations may have similar effects with phosphorylation by Prp4 kinase. In FgPrp31, the non-sense mutation at R464 resulted in the truncation of the C-terminal 130 aa region that contains all the conserved Prp4-phosphorylation sites. Deletion analysis showed that the N-terminal 310-aa rich in SR residues plays a critical role in the localization and functions of FgPrp4. We also conducted phosphoproteomics analysis with FgPrp4 and identified S289 as the phosphorylation site that is essential for its functions. These results indicated that FgPrp4 is critical for splicing efficiency but not essential for intron splicing, and FgPrp4 may regulate pre-mRNA splicing by phosphorylation of other components of the tri-snRNP although itself may be activated by phosphorylation at S289.

  1. Microtubule integrity regulates src-like and extracellular signal-regulated kinase activities in human pro-monocytic cells. Importance for interleukin-1 production.

    PubMed

    Schmid-Alliana, A; Menou, L; Manié, S; Schmid-Antomarchi, H; Millet, M A; Giuriato, S; Ferrua, B; Rossi, B

    1998-02-01

    We have demonstrated previously that microtubule depolymerization by colchicine in human monocytes induces selective production of interleukin-1 (IL-1) (Manié, S., Schmid-Alliana, A., Kubar, J., Ferrua, B., and Rossi, B. (1993) J. Biol. Chem. 268, 13675-13681). Here, we provide evidence that disruption of the microtubule structure rapidly triggers extracellular signal-regulated kinase (ERK) activation, whereas it was without effect on SAPK2 activity, which is commonly acknowledged to control pro-inflammatory cytokine production. This process involves the activation of the entire cascade including Ras, Raf-1, MEK1/2, ERK1, and ERK2. Activation of ERKs is followed by their nuclear translocation. Although other SAPK congeners might be activated upon microtubule depolymerization, the activation of ERK1 and ERK2 is mandatory for IL-1 production as shown by the blocking effect of PD 98059, a specific MEK1/2 inhibitor. Additionally, we provide evidence that microtubule disruption also induces the activation of c-Src and Hck activities. The importance of Src kinases in the mediation of the colchicine effect is underscored by the fact that CP 118556, a specific inhibitor of Src-like kinase, abrogates both the colchicine-induced ERK activation and IL-1 production. This is the first evidence that ERK activation is an absolute prerequisite for induction of this cytokine. Altogether, our data lend support to a model where the status of microtubule integrity controls the level of Src activities that subsequently activate the ERK kinase cascade, thus leading to IL-1 production.

  2. A Brief Review of Research Using Near-Infrared Spectroscopy to Measure Activation of the Prefrontal Cortex during Emotional Processing: The Importance of Experimental Design

    PubMed Central

    Bendall, Robert C. A.; Eachus, Peter; Thompson, Catherine

    2016-01-01

    During the past two decades there has been a pronounced increase in the number of published research studies that have employed near-infrared spectroscopy (NIRS) to measure neural activation. The technique is now an accepted neuroimaging tool adopted by cognitive neuroscientists to investigate a number of fields, one of which is the study of emotional processing. Crucially, one brain region that is important to the processing of emotional information is the prefrontal cortex (PFC) and NIRS is ideally suited to measuring activity in this region. Compared to other methods used to record neural activation, NIRS reduces the discomfort to participants, makes data collection from larger sample sizes more achievable, and allows measurement of activation during tasks involving physical movement. However, the use of NIRS to investigate the links between emotion and cognition has revealed mixed findings. For instance, whilst some studies report increased PFC activity associated with the processing of negative information, others show increased activity in relation to positive information. Research shows differences in PFC activity between different cognitive tasks, yet findings also vary within similar tasks. This work reviews a selection of recent studies that have adopted NIRS to study PFC activity during emotional processing in both healthy individuals and patient populations. It highlights the key differences between research findings and argues that variations in experimental design could be a contributing factor to the mixed results. Guidance is provided for future work in this area in order to improve consistency within this growing field. PMID:27812329

  3. Uncovering an Important Role for YopJ in the Inhibition of Caspase-1 in Activated Macrophages and Promoting Yersinia pseudotuberculosis Virulence.

    PubMed

    Schoberle, Taylor J; Chung, Lawton K; McPhee, Joseph B; Bogin, Ben; Bliska, James B

    2016-04-01

    Pathogenic Yersinia species utilize a type III secretion system to translocate Yop effectors into infected host cells. Yop effectors inhibit innate immune responses in infected macrophages to promote Yersinia pathogenesis. In turn,Yersinia-infected macrophages respond to translocation of Yops by activating caspase-1, but different mechanisms of caspase-1 activation occur, depending on the bacterial genotype and the state of phagocyte activation. In macrophages activated with lipopolysaccharide (LPS) prior to Yersinia pseudotuberculosis infection, caspase-1 is activated by a rapid inflammasome-dependent mechanism that is inhibited by translocated YopM. The possibility that other effectors cooperate with YopM to inhibit caspase-1 activation in LPS-activated macrophages has not been investigated. Toward this aim, epistasis analysis was carried out in which the phenotype of aY. pseudotuberculosis yopM mutant was compared to that of a yopJ yopM, yopE yopM, yopH yopM, yopT yopM, or ypkA yopM mutant. Activation of caspase-1 was measured by cleavage of the enzyme, release of interleukin-1β (IL-1β), and pyroptosis in LPS-activated macrophages infected with wild-type or mutant Y. pseudotuberculosis strains. Results show enhanced activation of caspase-1 after infection with the yopJ yopM mutant relative to infection by any other single or double mutant. Similar results were obtained with the yopJ, yopM, and yopJ yopM mutants ofY ersinia pestis Following intravenous infection of mice, theY. pseudotuberculosis yopJ mutant was as virulent as the wild type, while the yopJ yopM mutant was significantly more attenuated than the yopM mutant. In summary, through epistasis analysis this work uncovered an important role for YopJ in inhibiting caspase-1 in activated macrophages and in promoting Yersinia virulence.

  4. The second-sphere residue T263 is important for the function and catalytic activity of PTP1B via interaction with the WPD-loop.

    PubMed

    Xiao, Peng; Wang, Xiao; Wang, Hong-Mei; Fu, Xiao-Lei; Cui, Fu-ai; Yu, Xiao; Wen, Shi-shuai; Bi, Wen-Xiang; Sun, Jin-Peng

    2014-12-01

    Protein tyrosine phosphatases have diverse substrate specificities and intrinsic activities that lay the foundations for the fine-tuning of a phosphorylation network to precisely regulate cellular signal transduction. All classical PTPs share common catalytic mechanisms, and the important catalytic residues in the first sphere of their active sites have been well characterized. However, little attention has been paid to the second-sphere residues that are potentially important in defining the intrinsic activity and substrate specificity of PTPs. Here, we find that a conserved second-sphere residue, Thr263, located in the surface Q-loop is important for both the function and activity of PTPs. Using PTP1B as a study model, we found that mutations of Thr263 impaired the negative regulation role of PTP1B in insulin signaling. A detailed mechanistic study utilizing steady-state kinetics, Brønsted analysis and pH dependence in the presence of pNPP or phosphopeptide substrates revealed that Thr263 is required for the stabilization of the leaving group during catalysis. Further crystallographic studies and structural comparison revealed that Thr263 regulates the general acid function through modulation of the WPD-loop by the T263:F182/Y/H interaction pair, which is conserved in 26 out of 32 classical PTPs. In addition, the hydrophobic interaction between Thr263 and Arg1159 of the insulin receptor contributes to the substrate specificity of PTP1B. Taken together, our findings demonstrate the general role of the second-sphere residue Thr263 in PTP catalysis. Our findings suggest that the second sphere residues of PTP active site may play important roles in PTP-mediated function in both normal and diseased states. PMID:25450460

  5. Testing the impact of virus importation rates and future climate change on dengue activity in Malaysia using a mechanistic entomology and disease model.

    PubMed

    Williams, C R; Gill, B S; Mincham, G; Mohd Zaki, A H; Abdullah, N; Mahiyuddin, W R W; Ahmad, R; Shahar, M K; Harley, D; Viennet, E; Azil, A; Kamaluddin, A

    2015-10-01

    We aimed to reparameterize and validate an existing dengue model, comprising an entomological component (CIMSiM) and a disease component (DENSiM) for application in Malaysia. With the model we aimed to measure the effect of importation rate on dengue incidence, and to determine the potential impact of moderate climate change (a 1 °C temperature increase) on dengue activity. Dengue models (comprising CIMSiM and DENSiM) were reparameterized for a simulated Malaysian village of 10 000 people, and validated against monthly dengue case data from the district of Petaling Jaya in the state of Selangor. Simulations were also performed for 2008-2012 for variable virus importation rates (ranging from 1 to 25 per week) and dengue incidence determined. Dengue incidence in the period 2010-2012 was modelled, twice, with observed daily weather and with a 1 °C increase, the latter to simulate moderate climate change. Strong concordance between simulated and observed monthly dengue cases was observed (up to r = 0·72). There was a linear relationship between importation and incidence. However, a doubling of dengue importation did not equate to a doubling of dengue activity. The largest individual dengue outbreak was observed with the lowest dengue importation rate. Moderate climate change resulted in an overall decrease in dengue activity over a 3-year period, linked to high human seroprevalence early on in the simulation. Our results suggest that moderate reductions in importation with control programmes may not reduce the frequency of large outbreaks. Moderate increases in temperature do not necessarily lead to greater dengue incidence. PMID:25591942

  6. Testing the impact of virus importation rates and future climate change on dengue activity in Malaysia using a mechanistic entomology and disease model.

    PubMed

    Williams, C R; Gill, B S; Mincham, G; Mohd Zaki, A H; Abdullah, N; Mahiyuddin, W R W; Ahmad, R; Shahar, M K; Harley, D; Viennet, E; Azil, A; Kamaluddin, A

    2015-10-01

    We aimed to reparameterize and validate an existing dengue model, comprising an entomological component (CIMSiM) and a disease component (DENSiM) for application in Malaysia. With the model we aimed to measure the effect of importation rate on dengue incidence, and to determine the potential impact of moderate climate change (a 1 °C temperature increase) on dengue activity. Dengue models (comprising CIMSiM and DENSiM) were reparameterized for a simulated Malaysian village of 10 000 people, and validated against monthly dengue case data from the district of Petaling Jaya in the state of Selangor. Simulations were also performed for 2008-2012 for variable virus importation rates (ranging from 1 to 25 per week) and dengue incidence determined. Dengue incidence in the period 2010-2012 was modelled, twice, with observed daily weather and with a 1 °C increase, the latter to simulate moderate climate change. Strong concordance between simulated and observed monthly dengue cases was observed (up to r = 0·72). There was a linear relationship between importation and incidence. However, a doubling of dengue importation did not equate to a doubling of dengue activity. The largest individual dengue outbreak was observed with the lowest dengue importation rate. Moderate climate change resulted in an overall decrease in dengue activity over a 3-year period, linked to high human seroprevalence early on in the simulation. Our results suggest that moderate reductions in importation with control programmes may not reduce the frequency of large outbreaks. Moderate increases in temperature do not necessarily lead to greater dengue incidence.

  7. 2'-deoxy cyclic adenosine 5'-diphosphate ribose derivatives: importance of the 2'-hydroxyl motif for the antagonistic activity of 8-substituted cADPR derivatives.

    PubMed

    Zhang, Bo; Wagner, Gerd K; Weber, Karin; Garnham, Clive; Morgan, Anthony J; Galione, Antony; Guse, Andreas H; Potter, Barry V L

    2008-03-27

    The structural features needed for antagonism at the cyclic ADP-ribose (cADPR) receptor are unclear. Chemoenzymatic syntheses of novel 8-substituted 2'-deoxy-cADPR analogues, including 8-bromo-2'-deoxy-cADPR 7, 8-amino-2'-deoxy-cADPR 8, 8- O-methyl-2'-deoxy-cADPR 9, 8-phenyl-2'-deoxy-cADPR 10 and its ribose counterpart 8-phenyl-cADPR 5 are reported, including improved syntheses of established antagonists 8-amino-cADPR 2 and 8-bromo-cADPR 3. Aplysia californica ADP-ribosyl cyclase tolerates even the bulky 8-phenyl-nicotinamide adenine 5'-dinucleotide as a substrate. Structure-activity relationships of 8-substituted cADPR analogues in both Jurkat T-lymphocytes and sea urchin egg homogenate (SUH) were investigated. 2'-OH Deletion decreased antagonistic activity (at least for the 8-amino series), showing it to be an important motif. Some 8-substituted 2'-deoxy analogues showed agonist activity at higher concentrations, among which 8-bromo-2'-deoxy-cADPR 7 was, unexpectedly, a weak but almost full agonist in SUH and was membrane-permeant in whole eggs. Classical antagonists 2 and 3 also showed previously unobserved agonist activity at higher concentrations in both systems. The 2'-OH group, without effect on the Ca (2+)-mobilizing ability of cADPR itself, is an important motif for the antagonistic activities of 8-substituted cADPR analogues. PMID:18303825

  8. TGF-β-Smad2 dependent activation of CDC 25A plays an important role in cell proliferation through NFAT activation in metastatic breast cancer cells.

    PubMed

    Sengupta, Suman; Jana, Samir; Bhattacharyya, Arindam

    2014-02-01

    In late stages of cancer, TGF-β promotes the metastasis process by enhancing the invasiveness of cancer cells and inducing the epithelial-to-mesenchymal transition (EMT), a process that is concomitantly associated with breast cancer metastasis. Metastasis comprises of multiple steps with the regulation of complex network of signaling. Metastasis is associated with both the EMT and cell proliferation, but yet it has not been clearly distinguished how the balance between the cell proliferation and EMT is maintained together. Recently, it has been accounted that a transcription factor, NFAT has an important role for switching tumor suppressive to progressive effect of TGF-β and NFAT has a role in TGF-β mediated EMT by regulating N-cadherin. CDC 25A phosphatase, an important cell cycle regulator is overexpressed in breast cancer. Our results demonstrate that TGF-β regulating the CDC 25A in a Smad2 dependent way, translocates NFAT to nucleus and NFAT in co-operation with Smad2 promotes the tumor progression by upregulating the CDK2, CDK4, and cyclin E. This result signifies that TGF-β by regulating NFAT in different ways maintains the balance between EMT and cell proliferation mechanism concurrently during the late stage of breast cancer.

  9. Improving Hiroshima Air-Over-Ground Thermal/Epithermal Activation Calculations Using a MUSH Model to Show the Importance of Local Shielding

    SciTech Connect

    Pace, J.V.

    2002-02-14

    Achieving agreement between measured and calculated neutron activation data resulting from Hiroshima and Nagasaki A-bomb detonations has been a major problem since the early 1980's. This has been particularly true for the materials that are activated by thermal and epithermal neutrons. Since thermal and epithermal neutrons are not transported very far from the weapon, the local shielding environment around the measurement location can be very important. A set of calculations incorporating an average density local-environment material (mush) has been made to demonstrate that the local environment plays an important role in the calculation-measurement agreement process. The optimum solution would be to include the local environment in all thermal neutron response calculations.

  10. Stress imparted by the great 2004 Sumatra earthquake shut down transforms and activated rifts up to 400 km away in the Andaman Sea

    PubMed Central

    Sevilgen, Volkan; Stein, Ross S.; Pollitz, Fred F.

    2012-01-01

    The origin and prevalence of triggered seismicity and remote aftershocks are under debate. As a result, they have been excluded from probabilistic seismic hazard assessment and aftershock hazard notices. The 2004 M = 9.2 Sumatra earthquake altered seismicity in the Andaman backarc rift-transform system. Here we show that over a 300-km-long largely transform section of the backarc, M≥4.5 earthquakes stopped for five years, and over a 750-km-long backarc section, the rate of transform events dropped by two-thirds, while the rate of rift events increased eightfold. We compute the propagating dynamic stress wavefield and find the peak dynamic Coulomb stress is similar on the rifts and transforms. Long-period dynamic stress amplitudes, which are thought to promote dynamic failure, are higher on the transforms than on the rifts, opposite to the observations. In contrast to the dynamic stress, we calculate that the mainshock brought the transform segments approximately 0.2 bar (0.02 MPa) farther from static Coulomb failure and the rift segments approximately 0.2 bar closer to static failure, consistent with the seismic observations. This accord means that changes in seismicity rate are sufficiently predictable to be included in post-mainshock hazard evaluations. PMID:22949694

  11. Stress imparted by the great 2004 Sumatra earthquake shut down transforms and activated rifts up to 400 km away in the Andaman Sea

    USGS Publications Warehouse

    Sevilgen, Volkan; Stein, Ross S.; Pollitz, Fred F.

    2012-01-01

    The origin and prevalence of triggered seismicity and remote aftershocks are under debate. As a result, they have been excluded from probabilistic seismic hazard assessment and aftershock hazard notices. The 2004 M = 9.2 Sumatra earthquake altered seismicity in the Andaman backarc rift-transform system. Here we show that over a 300-km-long largely transform section of the backarc, M ≥ 4.5 earthquakes stopped for five years, and over a 750-km-long backarc section, the rate of transform events dropped by two-thirds, while the rate of rift events increased eightfold. We compute the propagating dynamic stress wavefield and find the peak dynamic Coulomb stress is similar on the rifts and transforms. Long-period dynamic stress amplitudes, which are thought to promote dynamic failure, are higher on the transforms than on the rifts, opposite to the observations. In contrast to the dynamic stress, we calculate that the mainshock brought the transform segments approximately 0.2 bar (0.02 MPa) farther from static Coulomb failure and the rift segments approximately 0.2 bar closer to static failure, consistent with the seismic observations. This accord means that changes in seismicity rate are sufficiently predictable to be included in post-mainshock hazard evaluations.

  12. Stress imparted by the great 2004 Sumatra earthquake shut down transforms and activated rifts up to 400 km away in the Andaman Sea.

    PubMed

    Sevilgen, Volkan; Stein, Ross S; Pollitz, Fred F

    2012-09-18

    The origin and prevalence of triggered seismicity and remote aftershocks are under debate. As a result, they have been excluded from probabilistic seismic hazard assessment and aftershock hazard notices. The 2004 M = 9.2 Sumatra earthquake altered seismicity in the Andaman backarc rift-transform system. Here we show that over a 300-km-long largely transform section of the backarc, M≥4.5 earthquakes stopped for five years, and over a 750-km-long backarc section, the rate of transform events dropped by two-thirds, while the rate of rift events increased eightfold. We compute the propagating dynamic stress wavefield and find the peak dynamic Coulomb stress is similar on the rifts and transforms. Long-period dynamic stress amplitudes, which are thought to promote dynamic failure, are higher on the transforms than on the rifts, opposite to the observations. In contrast to the dynamic stress, we calculate that the mainshock brought the transform segments approximately 0.2 bar (0.02 MPa) farther from static Coulomb failure and the rift segments approximately 0.2 bar closer to static failure, consistent with the seismic observations. This accord means that changes in seismicity rate are sufficiently predictable to be included in post-mainshock hazard evaluations.

  13. IPO3-mediated Nonclassical Nuclear Import of NF-κB Essential Modulator (NEMO) Drives DNA Damage-dependent NF-κB Activation*

    PubMed Central

    Hwang, Byounghoon; McCool, Kevin; Wan, Jun; Wuerzberger-Davis, Shelly M.; Young, Edmond W. K.; Choi, Eun Young; Cingolani, Gino; Weaver, Beth A.; Miyamoto, Shigeki

    2015-01-01

    Activation of IκB kinase (IKK) and NF-κB by genotoxic stresses modulates apoptotic responses and production of inflammatory mediators, thereby contributing to therapy resistance and premature aging. We previously reported that genotoxic agents induce nuclear localization of NF-κB essential modulator (NEMO) via an undefined mechanism to arbitrate subsequent DNA damage-dependent IKK/NF-κB signaling. Here we show that a nonclassical nuclear import pathway via IPO3 (importin 3, transportin 2) mediates stress-induced NEMO nuclear translocation. We found putative nuclear localization signals in NEMO whose mutations disrupted stress-inducible nuclear translocation of NEMO and IKK/NF-κB activation in stably reconstituted NEMO-deficient cells. RNAi screening of both importin α and β family members, as well as co-immunoprecipitation analyses, revealed that a nonclassical importin β family member, IPO3, was the only importin that was able to associate with NEMO and whose reduced expression prevented genotoxic stress-induced NEMO nuclear translocation, IKK/NF-κB activation, and inflammatory cytokine transcription. Recombinant IPO3 interacted with recombinant NEMO but not the nuclear localization signal mutant version and induced nuclear import of NEMO in digitonin-permeabilized cells. We also provide evidence that NEMO is disengaged from IKK complex following genotoxic stress induction. Thus, the IPO3 nuclear import pathway is an early and crucial determinant of the IKK/NF-κB signaling arm of the mammalian DNA damage response. PMID:26060253

  14. IPO3-mediated Nonclassical Nuclear Import of NF-κB Essential Modulator (NEMO) Drives DNA Damage-dependent NF-κB Activation.

    PubMed

    Hwang, Byounghoon; McCool, Kevin; Wan, Jun; Wuerzberger-Davis, Shelly M; Young, Edmond W K; Choi, Eun Young; Cingolani, Gino; Weaver, Beth A; Miyamoto, Shigeki

    2015-07-17

    Activation of IκB kinase (IKK) and NF-κB by genotoxic stresses modulates apoptotic responses and production of inflammatory mediators, thereby contributing to therapy resistance and premature aging. We previously reported that genotoxic agents induce nuclear localization of NF-κB essential modulator (NEMO) via an undefined mechanism to arbitrate subsequent DNA damage-dependent IKK/NF-κB signaling. Here we show that a nonclassical nuclear import pathway via IPO3 (importin 3, transportin 2) mediates stress-induced NEMO nuclear translocation. We found putative nuclear localization signals in NEMO whose mutations disrupted stress-inducible nuclear translocation of NEMO and IKK/NF-κB activation in stably reconstituted NEMO-deficient cells. RNAi screening of both importin α and β family members, as well as co-immunoprecipitation analyses, revealed that a nonclassical importin β family member, IPO3, was the only importin that was able to associate with NEMO and whose reduced expression prevented genotoxic stress-induced NEMO nuclear translocation, IKK/NF-κB activation, and inflammatory cytokine transcription. Recombinant IPO3 interacted with recombinant NEMO but not the nuclear localization signal mutant version and induced nuclear import of NEMO in digitonin-permeabilized cells. We also provide evidence that NEMO is disengaged from IKK complex following genotoxic stress induction. Thus, the IPO3 nuclear import pathway is an early and crucial determinant of the IKK/NF-κB signaling arm of the mammalian DNA damage response.

  15. Leptin signalling and leptin-mediated activation of human platelets: importance of JAK2 and the phospholipases Cgamma2 and A2.

    PubMed

    Dellas, Claudia; Schäfer, Katrin; Rohm, Ilonka K; Lankeit, Mareike; Leifheit, Maren; Loskutoff, David J; Hasenfuss, Gerd; Konstantinides, Stavros V

    2007-11-01

    Leptin enhances agonist-induced platelet aggregation, and human platelets have been reported to express the leptin receptor. However, the pathways and mediators lying downstream of leptin binding to platelets remain, with few exceptions, unknown. In the present study, we sought to gain further insight into the possible role of leptin as a platelet agonist. Stimulation of platelets with leptin promoted thromboxane generation and activation of alpha(IIb)beta(3), as demonstrated by PAC-1 binding. Furthermore, it increased the adhesion to immobilised fibrinogen (p<0.001) and induced cytoskeletal rearrangement of both platelets and Meg01 cells. Leptin time- and dose-dependently phosphorylated the intracellular signalling molecules JAK2 and STAT3, although the importance of STAT3 for leptin-induced platelet activation remains to be determined. Important intracellular mediators and pathways activated by leptin downstream of JAK2 were found to include phosphatidylinositol-3 kinase, phospholipase Cgamma2 and protein kinase C, as well as the p38 MAP kinase-phospholipase A(2) axis. Accordingly, incubation with the specific inhibitors AG490, Ly294002, U73122, and SB203580 prevented leptin-mediated platelet activation. These results help delineate biologically relevant leptin signalling pathways in platelets and may improve our understanding of the mechanisms linking hyperleptinaemia to the increased thrombosis risk in human obesity. PMID:18000612

  16. The nuclear protein GmbZIP110 has transcription activation activity and plays important roles in the response to salinity stress in soybean

    PubMed Central

    Xu, Zhaolong; Ali, Zulfiqar; Xu, Ling; He, Xiaolan; Huang, Yihong; Yi, Jinxin; Shao, Hongbo; Ma, Hongxiang; Zhang, Dayong

    2016-01-01

    Plant basic-leucine zipper (bZIP) transcription factors play important roles in many biological processes and are involved in the regulation of salt stress tolerance. Previously, our lab generated digital gene expression profiling (DGEP) data to identify differentially expressed genes in a salt-tolerant genotype of Glycine soja (STGoGS) and a salt-sensitive genotype of Glycine max (SSGoGM). This DGEP data revealed that the expression (log2 ratio) of GmbZIP110 was up-regulated 2.76-fold and 3.38-fold in SSGoGM and STGoGS, respectively. In the present study, the salt inducible gene GmbZIP110 was cloned and characterized through phylogenetic analysis, subcellular localization and in silico transcript abundance analysis in different tissues. The functional role of this gene in salt tolerance was studied through transactivation analysis, DNA binding ability, expression in soybean composite seedlings and transgenic Arabidopsis, and the effect of GmbZIP110 on the expression of stress-related genes in transgenic Arabidopsis was investigated. We found that GmbZIP110 could bind to the ACGT motif, impact the expression of many stress-related genes and the accumulation of proline, Na+ and K+, and enhanced the salt tolerance of composite seedlings and transgenic Arabidopsis. Integrating all these results, we propose that GmbZIP110 plays a critical role in the response to salinity stress in soybean and has high potential usefulness in crop improvement. PMID:26837841

  17. Membrane cholesterol plays an important role in enteropathogen adhesion and the activation of innate immunity via flagellin-TLR5 signaling.

    PubMed

    Zhou, Mingxu; Duan, Qiangde; Li, Yinchau; Yang, Yang; Hardwidge, Philip R; Zhu, Guoqiang

    2015-08-01

    Lipid rafts are cholesterol- and sphingolipid-rich ordered microdomains distributed in the plasma membrane that participates in mammalian signal transduction pathways. To determine the role of lipid rafts in mediating interactions between enteropathogens and intestinal epithelial cells, membrane cholesterol was depleted from Caco-2 and IPEC-J2 cells using methyl-β-cyclodextrin. Cholesterol depletion significantly reduced Escherichia coli and Salmonella enteritidis adhesion and invasion into intestinal epithelial cells. Complementation with exogenous cholesterol restored bacterial adhesion to basal levels. We also evaluated the role of lipid rafts in the activation of Toll-like receptor 5 signaling by bacterial flagellin. Depleting membrane cholesterol reduced the ability of purified recombinant E. coli flagellin to activate TLR5 signaling in intestinal cells. These data suggest that both membrane cholesterol and lipid rafts play important roles in enteropathogen adhesion and contribute to the activation of innate immunity via flagellin-TLR5 signaling.

  18. The N-terminal hybrid binding domain of RNase HI from Thermotoga maritima is important for substrate binding and Mg2+-dependent activity.

    PubMed

    Jongruja, Nujarin; You, Dong-Ju; Kanaya, Eiko; Koga, Yuichi; Takano, Kazufumi; Kanaya, Shigenori

    2010-11-01

    Thermotoga maritima ribonuclease H (RNase H) I (Tma-RNase HI) contains a hybrid binding domain (HBD) at the N-terminal region. To analyze the role of this HBD, Tma-RNase HI, Tma-W22A with the single mutation at the HBD, the C-terminal RNase H domain (Tma-CD) and the N-terminal domain containing the HBD (Tma-ND) were overproduced in Escherichia coli, purified and biochemically characterized. Tma-RNase HI prefers Mg(2+) to Mn(2+) for activity, and specifically loses most of the Mg(2+)-dependent activity on removal of the HBD and 87% of it by the mutation at the HBD. Tma-CD lost the ability to suppress the RNase H deficiency of an E. coli rnhA mutant, indicating that the HBD is responsible for in vivo RNase H activity. The cleavage-site specificities of Tma-RNase HI are not significantly changed on removal of the HBD, regardless of the metal cofactor. Binding analyses of the proteins to the substrate using surface plasmon resonance indicate that the binding affinity of Tma-RNase HI is greatly reduced on removal of the HBD or the mutation. These results indicate that there is a correlation between Mg(2+)-dependent activity and substrate binding affinity. Tma-CD was as stable as Tma-RNase HI, indicating that the HBD is not important for stability. The HBD of Tma-RNase HI is important not only for substrate binding, but also for Mg(2+)-dependent activity, probably because the HBD affects the interaction between the substrate and enzyme at the active site, such that the scissile phosphate group of the substrate and the Mg(2+) ion are arranged ideally.

  19. Mutations in Mtr4 Structural Domains Reveal Their Important Role in Regulating tRNAiMet Turnover in Saccharomyces cerevisiae and Mtr4p Enzymatic Activities In Vitro.

    PubMed

    Li, Yan; Burclaff, Joseph; Anderson, James T

    2016-01-01

    RNA processing and turnover play important roles in the maturation, metabolism and quality control of a large variety of RNAs thereby contributing to gene expression and cellular health. The TRAMP complex, composed of Air2p, Trf4p and Mtr4p, stimulates nuclear exosome-dependent RNA processing and degradation in Saccharomyces cerevisiae. The Mtr4 protein structure is composed of a helicase core and a novel so-called arch domain, which protrudes from the core. The helicase core contains highly conserved helicase domains RecA-1 and 2, and two structural domains of unclear functions, winged helix domain (WH) and ratchet domain. How the structural domains (arch, WH and ratchet domain) coordinate with the helicase domains and what roles they are playing in regulating Mtr4p helicase activity are unknown. We created a library of Mtr4p structural domain mutants for the first time and screened for those defective in the turnover of TRAMP and exosome substrate, hypomodified tRNAiMet. We found these domains regulate Mtr4p enzymatic activities differently through characterizing the arch domain mutants K700N and P731S, WH mutant K904N, and ratchet domain mutant R1030G. Arch domain mutants greatly reduced Mtr4p RNA binding, which surprisingly did not lead to significant defects on either in vivo tRNAiMet turnover, or in vitro unwinding activities. WH mutant K904N and Ratchet domain mutant R1030G showed decreased tRNAiMet turnover in vivo, as well as reduced RNA binding, ATPase and unwinding activities of Mtr4p in vitro. Particularly, K904 was found to be very important for steady protein levels in vivo. Overall, we conclude that arch domain plays a role in RNA binding but is largely dispensable for Mtr4p enzymatic activities, however the structural domains in the helicase core significantly contribute to Mtr4p ATPase and unwinding activities.

  20. Fractionating ambient humic-like substances (HULIS) for their reactive oxygen species activity - Assessing the importance of quinones and atmospheric aging

    NASA Astrophysics Data System (ADS)

    Verma, Vishal; Wang, Ying; El-Afifi, Rawan; Fang, Ting; Rowland, Janessa; Russell, Armistead G.; Weber, Rodney J.

    2015-11-01

    In this paper, we present a technique to identify the redox-active components of fine organic aerosols by fractionating humic-like substances (HULIS). We applied this technique to a dithiothreitol (DTT) assay - a measure of the capability of PM to generate reactive oxygen species (ROS), and assessed the contribution of quinones to the DTT activity of ambient water-soluble PM. Filter samples from the Southeastern Center for Air Pollution & Epidemiology (SCAPE) were extracted in water and then passed-through a C-18 column to isolate the HULIS fraction by retention on the column. The HULIS was then eluted with a sequence of solvents of increasing polarity, i.e., hexane, dichloromethane (DCM) and then methanol. Each of these eluted fractions was analyzed for DTT activity. The methanol fraction was found to possess most of the DTT activity (>70%), while the hexane fraction had the least activity (<5%), suggesting that the ROS-active compounds of ambient water-soluble PM2.5 HULIS are mostly polar in nature. A number of quinones thought to contribute to ambient PM DTT activity were also tested. 1,4 Naphthoquinone (1,4 NQ), 1,2 Naphthoquinone (1,2 NQ), 9,10 Phenanthrenequinone (PQ), and 5-hydroxy-1,4 NQ were analyzed by the same protocol. The hexane fraction of two quinones (PQ, and 1,4 NQ) was the most-DTT active, while methanol was the least, confirming that PQ, 1,4 NQ, and 1,2 NQ (which could not be recovered from the column) do not contribute significantly to the water-soluble DTT activity of ambient PM2.5. However, an oxygenated derivative of 1,4 NQ, (5-hydroxy-1,4 NQ), which is also intrinsically more DTT-active than 1,4 NQ, was mostly (>60%) eluted in methanol. The results demonstrate the importance of atmospheric aging (oxidation) of organic aerosols in enhancing the ROS activity of ambient PM.

  1. Motor patterns of the small intestine explained by phase-amplitude coupling of two pacemaker activities: the critical importance of propagation velocity.

    PubMed

    Huizinga, Jan D; Parsons, Sean P; Chen, Ji-Hong; Pawelka, Andrew; Pistilli, Marc; Li, Chunpei; Yu, Yuanjie; Ye, Pengfei; Liu, Qing; Tong, Mengting; Zhu, Yong Fang; Wei, Defei

    2015-09-15

    Phase-amplitude coupling of two pacemaker activities of the small intestine, the omnipresent slow wave activity generated by interstitial cells of Cajal of the myenteric plexus (ICC-MP) and the stimulus-dependent rhythmic transient depolarizations generated by ICC of the deep muscular plexus (ICC-DMP), was recently hypothesized to underlie the orchestration of the segmentation motor pattern. The aim of the present study was to increase our understanding of phase-amplitude coupling through modeling. In particular the importance of propagation velocity of the ICC-DMP component was investigated. The outcome of the modeling was compared with motor patterns recorded from the rat or mouse intestine from which propagation velocities within the different patterns were measured. The results show that the classical segmentation motor pattern occurs when the ICC-DMP component has a low propagation velocity (<0.05 cm/s). When the ICC-DMP component has a propagation velocity in the same order of magnitude as that of the slow wave activity (∼1 cm/s), cluster type propulsive activity occurs which is in fact the dominant propulsive activity of the intestine. Hence, the only difference between the generation of propagating cluster contractions and the Cannon-type segmentation motor pattern is the propagation velocity of the low-frequency component, the rhythmic transient depolarizations originating from the ICC-DMP. Importantly, the proposed mechanism explains why both motor patterns have distinct rhythmic waxing and waning of the amplitude of contractions. The hypothesis is brought forward that the velocity is modulated by neural regulation of gap junction conductance within the ICC-DMP network.

  2. Proteasome activity is important for replication recovery, CHK1 phosphorylation and prevention of G2 arrest after low-dose formaldehyde

    SciTech Connect

    Ortega-Atienza, Sara; Green, Samantha E.; Zhitkovich, Anatoly

    2015-07-15

    Formaldehyde (FA) is a human carcinogen with numerous sources of environmental and occupational exposures. This reactive aldehyde is also produced endogenously during metabolism of drugs and other processes. DNA–protein crosslinks (DPCs) are considered to be the main genotoxic lesions for FA. Accumulating evidence suggests that DPC repair in high eukaryotes involves proteolysis of crosslinked proteins. Here, we examined a role of the main cellular proteolytic machinery proteasomes in toxic responses of human lung cells to low FA doses. We found that transient inhibition of proteasome activity increased cytotoxicity and diminished clonogenic viability of FA-treated cells. Proteasome inactivation exacerbated suppressive effects of FA on DNA replication and increased the levels of the genotoxic stress marker γ-H2AX in normal human cells. A transient loss of proteasome activity in FA-exposed cells also caused delayed perturbations of cell cycle, which included G2 arrest and a depletion of S-phase populations at FA doses that had no effects in control cells. Proteasome activity diminished p53-Ser15 phosphorylation but was important for FA-induced CHK1 phosphorylation, which is a biochemical marker of DPC proteolysis in replicating cells. Unlike FA, proteasome inhibition had no effect on cell survival and CHK1 phosphorylation by the non-DPC replication stressor hydroxyurea. Overall, we obtained evidence for the importance of proteasomes in protection of human cells against biologically relevant doses of FA. Biochemically, our findings indicate the involvement of proteasomes in proteolytic repair of DPC, which removes replication blockage by these highly bulky lesions. - Highlights: • Proteasome inhibition enhances cytotoxicity of low-dose FA in human lung cells. • Active proteasomes diminish replication-inhibiting effects of FA. • Proteasome activity prevents delayed G2 arrest in FA-treated cells. • Proteasome inhibition exacerbates replication stress by FA in

  3. TALE1 from Xanthomonas axonopodis pv. manihotis acts as a transcriptional activator in plant cells and is important for pathogenicity in cassava plants.

    PubMed

    Castiblanco, Luisa F; Gil, Juliana; Rojas, Alejandro; Osorio, Daniela; Gutiérrez, Sonia; Muñoz-Bodnar, Alejandra; Perez-Quintero, Alvaro L; Koebnik, Ralf; Szurek, Boris; López, Camilo; Restrepo, Silvia; Verdier, Valérie; Bernal, Adriana J

    2013-01-01

    Many plant-pathogenic bacteria suppress pathogen-associated molecular pattern (PAMP)-triggered immunity by injecting effector proteins into the host cytoplasm during infection through the type III secretion system (TTSS). This type III secretome plays an important role in bacterial pathogenicity in susceptible hosts. Xanthomonas axonopodis pv. manihotis (Xam), the causal agent of cassava bacterial blight, injects several effector proteins into the host cell, including TALE1(Xam) . This protein is a member of the Transcriptional Activator-Like effector (TALE) protein family, formerly known as the AvrBs3/PthA family. TALE1(Xam) has 13.5 tandem repeats of 34 amino acids each, as well as two nuclear localization signals and an acidic activation domain at the C-terminus. In this work, we demonstrate the importance of TALE1(Xam) in the pathogenicity of Xam. We use versions of the gene that lack different domains in the protein in structure-function studies to show that the eukaryotic domains at the 3' end are critical for pathogenicity. In addition, we demonstrate that, similar to the characterized TALE proteins from other Xanthomonas species, TALE1(Xam) acts as a transcriptional activator in plant cells. This is the first report of the identification of a TALE in Xam, and contributes to our understanding of the pathogenicity mechanisms employed by this bacterium to colonize and cause disease in cassava. PMID:22947214

  4. Stress-triggered Activation of the Metalloprotease Oma1 Involves Its C-terminal Region and Is Important for Mitochondrial Stress Protection in Yeast*

    PubMed Central

    Bohovych, Iryna; Donaldson, Garrett; Christianson, Sara; Zahayko, Nataliya; Khalimonchuk, Oleh

    2014-01-01

    Functional integrity of mitochondria is critical for optimal cellular physiology. A suite of conserved mitochondrial proteases known as intramitochondrial quality control represents one of the mechanisms assuring normal mitochondrial function. We previously demonstrated that ATP-independent metalloprotease Oma1 mediates degradation of hypohemylated Cox1 subunit of cytochrome c oxidase and is active in cytochrome c oxidase-deficient mitochondria. Here we show that Oma1 is important for adaptive responses to various homeostatic insults and preservation of normal mitochondrial function under damage-eliciting conditions. Changes in membrane potential, oxidative stress, or chronic hyperpolarization lead to increased Oma1-mediated proteolysis. The stress-triggered induction of Oma1 proteolytic activity appears to be associated with conformational changes within the Oma1 homo-oligomeric complex, and these alterations likely involve C-terminal residues of the protease. Substitutions in the conserved C-terminal region of Oma1 impair its ability to form a labile proteolytically active complex in response to stress stimuli. We demonstrate that Oma1 genetically interacts with other inner membrane-bound quality control proteases. These findings indicate that yeast Oma1 is an important player in IM protein homeostasis and integrity by acting in concert with other intramitochondrial quality control components. PMID:24648523

  5. Highly Active and Selective Manganese C=O Bond Hydrogenation Catalysts: The Importance of the Multidentate Ligand, the Ancillary Ligands, and the Oxidation State.

    PubMed

    Kallmeier, Fabian; Irrgang, Torsten; Dietel, Thomas; Kempe, Rhett

    2016-09-19

    The replacement of expensive noble metals by earth-abundant transition metals is a central topic in catalysis. Herein, we introduce a highly active and selective homogeneous manganese-based C=O bond hydrogenation catalyst. Our catalyst has a broad substrate scope, it is able to hydrogenate aryl-alkyl, diaryl, dialkyl, and cycloalkyl ketones as well as aldehydes. A very good functional group tolerance including the quantitative and selective hydrogenation of a ketone in the presence of a non-shielded olefin is observed. In Mn hydrogenation catalysis, the combination of the multidentate ligand, the oxidation state of the metal, and the choice of the right ancillary ligand is crucial for high activity. This observation emphasizes an advantage and the importance of homogeneous catalysts in 3d-metal catalysis. For coordination compounds, fine-tuning of a complex coordination environment is easily accomplished in comparison to enzyme and/or heterogeneous catalysts. PMID:27571701

  6. Amino acid sequence of toxin XI of the scorpion Buthus occitanus tunetanus. Evidence of a mutation having an important effect upon neurotoxic activity.

    PubMed

    Sampieri, F; Habersetzer-Rochat, C; Martin, M F; Kopeyan, C; Rochat, H

    1987-02-01

    The complete amino acid sequence of toxin XI of the North African scorpion Buthus occitanus tunetanus has been elucidated by automatic sequencing of the reduced and alkylated toxin and of the peptides obtained after tryptic cleavage restricted to arginyl bonds. This toxin is structurally homologous to toxin II of Androctonus australis Hector, the most active among the alpha-toxins, but is far less potent, both in vivo and in vitro. This work points out 12 mutations, many of which are conservative. Nevertheless, the most striking difference is the replacement of the lysine residue at position 58, known to be important in the activity of AaH toxin II, by a valine residue. Thus, it seems that the presence of a positive charge at this location facilitates the interactions between the receptor on the sodium channel and the alpha-type toxins.

  7. Analytic Hierarchy Process to Define the Most Important Factors and Related Technologies for Empowering Elderly People in Taking an Active Role in their Health.

    PubMed

    Fico, G; Gaeta, E; Arredondo, M T; Pecchia, L

    2015-09-01

    Successful management of health conditions in older population is determined by strategic involvement of a professional team of careers and by empowering patients and their caregivers to take over a central role and responsibility in the daily management of condition. Identifying, structuring and ranking the most important needs related to these aspects could pave the way for improved strategies in designing systems and technological solutions supporting user empowerment. This paper presents the preliminary results of a study aiming to elicit these needs. Healthcare professionals, working together in the European and Innovation Partnership on Active and Healthy Ageing (EIP-AHA) initiative, have defined a set of needs and factors that have been organized in two hierarchies around the concepts of patient activation and proactive and prepared care team, defined in the Chronic Care Model. The two hierarchies have been mapped, by a team of experts in computer science, with technologies and solutions that could facilitate the achievement of the identified needs.

  8. 2-Arylbenzothiazole, benzoxazole and benzimidazole derivatives as fluorogenic substrates for the detection of nitroreductase and aminopeptidase activity in clinically important bacteria.

    PubMed

    Cellier, Marie; Fabrega, Olivier J; Fazackerley, Elizabeth; James, Arthur L; Orenga, Sylvain; Perry, John D; Salwatura, Vindhya L; Stanforth, Stephen P

    2011-05-01

    A series of 2-(2-nitrophenyl)benzothiazole 7, 2-(2-nitrophenyl)benzoxazole 10 and 2-(2-nitrophenyl)benzimidazole 13 derivatives have been synthesised and assessed as indicators of nitroreductase activity across a range of clinically important Gram negative and Gram positive bacteria. The majority of Gram negative bacteria produced strongly fluorescent colonies with substrates 7 and 10 whereas fluorescence production in Gram positive bacteria was less widespread. The l-alanine 16 and 19 and β-alanine 21 and 23 derivatives have been prepared from 2-(2-aminophenyl)benzothiazole 14 and 2-(2-aminophenyl)benzoxazole 17. These four compounds have been evaluated as indicators of aminopeptidase activity. The growth of Gram positive bacteria was generally inhibited by these substrates but fluorescent colonies were produced with the majority of Gram negative bacteria tested.

  9. Mapping of a Region of the PA-X Protein of Influenza A Virus That Is Important for Its Shutoff Activity.

    PubMed

    Oishi, Kohei; Yamayoshi, Seiya; Kawaoka, Yoshihiro

    2015-08-01

    Influenza A virus PA-X comprises an N-terminal PA endonuclease domain and a C-terminal PA-X-specific domain. PA-X reduces host and viral mRNA accumulation via its endonuclease function. Here, we found that the N-terminal 15 amino acids, particularly six basic amino acids, in the C-terminal PA-X-specific region are important for PA-X shutoff activity. These six basic amino acids enabled a PA deletion mutant to suppress protein expression at a level comparable to that of wild-type PA-X.

  10. Contrasted enzymatic cocktails reveal the importance of cellulases and hemicellulases activity ratios for the hydrolysis of cellulose in presence of xylans.

    PubMed

    Dondelinger, Eve; Aubry, Nathalie; Ben Chaabane, Fadhel; Cohen, Céline; Tayeb, Jean; Rémond, Caroline

    2016-03-01

    Various enzymatic cocktails were produced from two Trichoderma reesei strains, a cellulase hyperproducer strain and a strain with β-glucosidase activity overexpression. By using various carbon sources (lactose, glucose, xylose, hemicellulosic hydrolysate) for strains growth, contrasted enzymatic activities were obtained. The enzymatic cocktails presented various levels of efficiency for the hydrolysis of cellulose Avicel into glucose, in presence of xylans, or not. These latter were also hydrolyzed with different extents according to cocktails. The most efficient cocktails (TR1 and TR3) on Avicel were richer in filter paper activity (FPU) and presented a low ratio FPU/β-glucosidase activity. Cocktails TR2 and TR5 which were produced on the higher amount of hemicellulosic hydrolysate, possess both high xylanase and β-xylosidase activities, and were the most efficient for xylans hydrolysis. When hydrolysis of Avicel was conducted in presence of xylans, a decrease of glucose release occurred for all cocktails compared to hydrolysis of Avicel alone. Mixing TR1 and TR5 cocktails with two different ratios of proteins (1/1 and 1/4) resulted in a gain of efficiency for glucose release during hydrolysis of Avicel in presence of xylans compared to TR5 alone. Our results demonstrate the importance of combining hemicellulase and cellulase activities to improve the yields of glucose release from Avicel in presence of xylans. In this context, strategies involving enzymes production with carbon sources comprising mixed C5 and C6 sugars or combining different cocktails produced on C5 or on C6 sugars are of interest for processes developed in the context of lignocellulosic biorefinery. PMID:27001439

  11. Chaperone-like activities of different molecular forms of beta-casein. Importance of polarity of N-terminal hydrophilic domain.

    PubMed

    Yousefi, Reza; Shchutskaya, Yulia Y; Zimny, Jaroslaw; Gaudin, Jean-Charles; Moosavi-Movahedi, Ali A; Muronetz, Vladimir I; Zuev, Yuriy F; Chobert, Jean-Marc; Haertlé, Thomas

    2009-08-01

    As a member of intrinsically unstructured protein family, beta-casein (beta-CN) contains relatively high amount of prolyl residues, adopts noncompact and flexible structure and exhibits chaperone-like activity in vitro. Like many chaperones, native beta-CN does not contain cysteinyl residues and exhibits strong tendencies for self-association. The chaperone-like activities of three recombinant beta-CNs wild type (WT) beta-CN, C4 beta-CN (with cysteinyl residue in position 4) and C208 beta-CN (with cysteinyl residue in position 208), expressed and purified from E. coli, which, consequently, lack the phosphorylated residues, were examined and compared with that of native beta-CN using insulin and alcohol dehydrogenase as target/substrate proteins. The dimers (beta-CND) of C4-beta-CN and C208 beta-CN were also studied and their chaperone-like activities were compared with those of their monomeric forms. Lacking phosphorylation, WT beta-CN, C208 beta-CN, C4 beta-CN and C4 beta-CND exhibited significantly lower chaperone-like activities than native beta-CN. Dimerization of C208 beta-CN with two distal hydrophilic domains considerably improved its chaperone-like activity in comparison with its monomeric form. The obtained results demonstrate the significant role played by the polar contributions of phosphorylated residues and N-terminal hydrophilic domain as important functional elements in enhancing the chaperone-like activity of native beta-CN. (c) 2009 Wiley Periodicals, Inc. Biopolymers 91: 623-632, 2009.This article was originally published online as an accepted preprint. The "Published Online" date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com. PMID:19322774

  12. Important role for phylogenetically invariant PP2Acalpha active site and C-terminal residues revealed by mutational analysis in Saccharomyces cerevisiae.

    PubMed Central

    Evans, D R; Hemmings, B A

    2000-01-01

    PP2A is a central regulator of eukaryotic signal transduction. The human catalytic subunit PP2Acalpha functionally replaces the endogenous yeast enzyme, Pph22p, indicating a conservation of function in vivo. Therefore, yeast cells were employed to explore the role of invariant PP2Ac residues. The PP2Acalpha Y127N substitution abolished essential PP2Ac function in vivo and impaired catalysis severely in vitro, consistent with the prediction from structural studies that Tyr-127 mediates substrate binding and its side chain interacts with the key active site residues His-118 and Asp-88. The V159E substitution similarly impaired PP2Acalpha catalysis profoundly and may cause global disruption of the active site. Two conditional mutations in the yeast Pph22p protein, F232S and P240H, were found to cause temperature-sensitive impairment of PP2Ac catalytic function in vitro. Thus, the mitotic and cell lysis defects conferred by these mutations result from a loss of PP2Ac enzyme activity. Substitution of the PP2Acalpha C-terminal Tyr-307 residue by phenylalanine impaired protein function, whereas the Y307D and T304D substitutions abolished essential function in vivo. Nevertheless, Y307D did not reduce PP2Acalpha catalytic activity significantly in vitro, consistent with an important role for the C terminus in mediating essential protein-protein interactions. Our results identify key residues important for PP2Ac function and characterize new reagents for the study of PP2A in vivo. PMID:10978272

  13. The Importance of the Interaction of CheD with CheC and the Chemoreceptors Compared to Its Enzymatic Activity during Chemotaxis in Bacillus subtilis

    PubMed Central

    Allen, George M.; Walukiewicz, Hanna E.; Rao, Christopher V.; Ordal, George W.

    2012-01-01

    Bacillus subtilis use three systems for adaptation during chemotaxis. One of these systems involves two interacting proteins, CheC and CheD. CheD binds to the receptors and increases their ability to activate the CheA kinase. CheD also binds CheC, and the strength of this interaction is increased by phosphorylated CheY. CheC is believed to control the binding of CheD to the receptors in response to the levels of phosphorylated CheY. In addition to their role in adaptation, CheC and CheD also have separate enzymatic functions. CheC is a CheY phosphatase and CheD is a receptor deamidase. Previously, we demonstrated that CheC’s phosphatase activity plays a minor role in chemotaxis whereas its ability to bind CheD plays a major one. In the present study, we demonstrate that CheD’s deamidase activity also plays a minor role in chemotaxis whereas its ability to bind CheC plays a major one. In addition, we quantified the interaction between CheC and CheD using surface plasmon resonance. These results suggest that the most important features of CheC and CheD are not their enzymatic activities but rather their roles in adaptation. PMID:23226535

  14. Recognizing the importance of exposure-dose-response dynamics for ecotoxicity assessment: nitrofurazone-induced antioxidase activity and mRNA expression in model protozoan Euplotes vannus.

    PubMed

    Hong, Yazhen; Liu, Shuxing; Lin, Xiaofeng; Li, Jiqiu; Yi, Zhenzhen; Al-Rasheid, Khaled A S

    2015-06-01

    The equivocality of dose-response relationships has, in practice, hampered the application of biomarkers as a means to evaluate environmental risk, yet this important issue has not yet been fully recognized or explored. This paper evaluates the potential of antioxidant enzymes in the ciliated protozoan Euplotes vannus for use as biomarkers. Dose-response dynamics, together with both the enzyme activity and the gene expression of the antioxidant enzymes, superoxide dismutase, and glutathione peroxidase, were investigated when E. vannus were exposed to graded doses of nitrofurazone for several discrete durations. Mathematical models were explored to characterize the dose-response profiles and, specifically, to identify any equivocality in terms of endpoint. Significant differences were found in both enzyme activity and messenger RNA (mRNA) expression in the E. vannus treated with nitrofurazone, and the interactions between exposure dosage and duration were significant. Correlations between enzyme activity, mRNA expression, and nitrofurazone dose varied with exposure duration. Particularly, the dose-responses showed different dynamics depending on either endpoint or exposure duration. Our findings suggest that both the enzyme activity and the gene expression of the tested antioxidant enzymes can be used as biomarkers for ecotoxicological assessment on the premise of ascertaining appropriate dosage scope, exposure duration, endpoint, etc., which can be achieved by using dose-response dynamics.

  15. Does weather play an important role in the early nesting activity of colonial waterbirds? A case study in putrajaya wetlands, malaysia.

    PubMed

    Ismail, Ahmad; Rahman, Faid

    2013-08-01

    Environmental factors can play important roles in influencing waterbird communities. In particular, weather may have various biological and ecological impacts on the breeding activities of waterbirds, though most studies have investigated the effect of weather on the late stages of waterbird breeding (e.g., hatching rate, chick mortality). Conversely, the present study attempts to highlight the influence of weather on the early nesting activities of waterbirds by evaluating a recently established mixed-species colony in Putrajaya Wetlands, Malaysia. The results show that only rainfall and temperature have a significant influence on the species' nesting activities. Rainfall activity is significantly correlated with the Grey Heron's rate of establishment (rainfall: rs = 0.558, p = 0.03, n = 72) whereas both temperature and rainfall are associated with Painted Stork's nesting density (temperature: rs = 0.573, p = 0.013; rainfall: rs = -0.662, p = 0.03, n = 48). There is a possibility that variations in the rainfall and temperature provide a cue for the birds to initiate their nesting. Regardless, this paper addresses concerns on the limitations faced in the study and suggests long-term studies for confirmation.

  16. Activated Kupffer cells play an important role in intra-hepatic Th1-associated necro-inflammation in Concanavalin A-induced hepatic injury in mice.

    PubMed

    Morita, Atsuhiro; Itoh, Yoshito; Toyama, Tetsuya; Fujii, Hideki; Nishioji, Kenichi; Kirishima, Toshihiko; Makiyama, Akiko; Yamauchi, Norihito; Okanoue, Takeshi

    2003-10-01

    BACKGROUND/AIMS: To examine whether or not activated Kupffer cells play an important role in intra-hepatic Th1-associated necro-inflammation in Concanavalin A (Con A)-induced hepatic injury in mice. METHODS: Con A was administered to Balb/c mice pretreated with or without gadolinium chloride (GdCl(3)). Kupffer cell activation was evaluated by their ability to produce superoxide anions in situ under liver perfusion with nitro blue tetrazolium (NBT). Hepatic concentration of cytokines was measured by ELISA and the mRNA expression of CXC chemokine receptor 3 (CXCR3) was evaluated by RT-PCR. Immunohistochemical detection of CD4 positive lymphocytes in the liver was also performed. RESULTS: GdCl(3)-pretreatment significantly (P<0.01) reduced the serum levels of alanine aminotransferase (ALT) in Con A-treated mice. Formazan deposition in Kupffer cells, the hepatic concentration of tumor necrosis factor-alpha and interferon-gamma, the mRNA expression of CXCR3 and the CD4 positive lymphocytes in the liver were decreased in GdCl(3)-pretreated mice as compared with those without GdCl(3)-pretreatment (P<0.05, respectively). CONCLUSIONS: Activated Kupffer cells, which produce superoxide anions, are involved in Con A-induced hepatic necro-inflammation in mice possibly through the activation of Th1-associated immune response mediated by CD4 and/or CXCR3 positive cells recruited into the liver.

  17. A Comprehensive Review on the Phytochemical Constituents and Pharmacological Activities of Pogostemon cablin Benth.: An Aromatic Medicinal Plant of Industrial Importance.

    PubMed

    Swamy, Mallappa Kumara; Sinniah, Uma Rani

    2015-01-01

    Pogostemon cablin Benth. (patchouli) is an important herb which possesses many therapeutic properties and is widely used in the fragrance industries. In traditional medicinal practices, it is used to treat colds, headaches, fever, nausea, vomiting, diarrhea, abdominal pain, insect and snake bites. In aromatherapy, patchouli oil is used to relieve depression, stress, calm nerves, control appetite and to improve sexual interest. Till now more than 140 compounds, including terpenoids, phytosterols, flavonoids, organic acids, lignins, alkaloids, glycosides, alcohols, aldehydes have been isolated and identified from patchouli. The main phytochemical compounds are patchouli alcohol, α-patchoulene, β-patchoulene, α-bulnesene, seychellene, norpatchoulenol, pogostone, eugenol and pogostol. Modern studies have revealed several biological activities such as antioxidant, analgesic, anti-inflammatory, antiplatelet, antithrombotic, aphrodisiac, antidepressant, antimutagenic, antiemetic, fibrinolytic and cytotoxic activities. However, some of the traditional uses need to be verified and may require standardizing and authenticating the bioactivity of purified compounds through scientific methods. The aim of the present review is to provide comprehensive knowledge on the phytochemistry and pharmacological activities of essential oil and different plant extracts of patchouli based on the available scientific literature. This information will provide a potential guide in exploring the use of main active compounds of patchouli in various medical fields. PMID:25985355

  18. Does Litter Impart A Detectable Chemical Signal on Soil DOC? DOC Fluorescence Signatures in Soils Undergoing Long-Term Litter Manipulations

    NASA Astrophysics Data System (ADS)

    Lajtha, K.; Strid, A.; Lee, B. S.

    2015-12-01

    Soil dissolved organic carbon (DOC) is a small but crucial part of the forest carbon cycle. Characterizing the relationship between organic matter inputs to soil and DOC chemistry is crucial to understanding the ultimate fate of root carbon, fallen wood and needles. Chemical differences in the DOC pool may help to explain whether fractions are sorbed to mineral surfaces and contribute to accumulation of soil organic carbon, respired as CO2, or exported. Soil solution DOC was sampled from the detrital input and removal treatment (DIRT) plots located in the H.J. Andrews Experimental Forest, OR to determine whether detrital inputs impart a detectable signal on DOC in mineral soil. Multiple types of fresh litter extracts, along with lysimeter and soil extracts from DIRT treatment plots were characterized using UV-Vis and fluorescence spectroscopy coupled with the Cory and McKnight (2005) parallel factor analysis (PARAFAC) model. Principal component analysis of 13 unique fluorophores distinguished using PARAFAC show that litter and soil extracts (needles, wood of decomposition Class 1, Class 3 and Class 5, O-horizon, and A-horizon) each have distinct fluorescence signatures. However, while litter-leached DOC chemistry varies by litter type, neither lysimeter-collected DOC or soil extracts show statistically significant differences in fluorescence signatures among treatments, even after 17 years of litter manipulations. The lack of observed differences among DIRT treatments suggests a "Soil Blender" hypothesis whereby both abiotic and biotic mechanisms effectively homogenize organic carbon constituents within the dissolved pool. The results of this work emphasize the ability of sorption and biodegradation to homogenize soil DOC and demonstrate that fluorescence can be an effective fingerprinting technique for soil DOC composition.

  19. CodY Regulates the Activity of the Virulence Quorum Sensor PlcR by Controlling the Import of the Signaling Peptide PapR in Bacillus thuringiensis

    PubMed Central

    Slamti, Leyla; Lemy, Christelle; Henry, Céline; Guillot, Alain; Huillet, Eugénie; Lereclus, Didier

    2016-01-01

    In Gram-positive bacteria, cell–cell communication mainly relies on cytoplasmic sensors of the RNPP family. Activity of these regulators depends on their binding to secreted signaling peptides that are imported into the cell. These quorum sensing regulators control important biological functions in bacteria of the Bacillus cereus group, such as virulence and necrotrophism. The RNPP quorum sensor PlcR, in complex with its cognate signaling peptide PapR, is the main regulator of virulence in B. cereus and Bacillus thuringiensis (Bt). Recent reports have shown that the global stationary phase regulator CodY, involved in adaptation to nutritional limitation, is required for the expression of virulence genes belonging to the PlcR regulon. However, the mechanism underlying this regulation was not described. Using genetics and proteomics approaches, we showed that CodY regulates the expression of the virulence genes through the import of PapR. We report that CodY positively controls the production of the proteins that compose the oligopeptide permease OppABCDF, and of several other Opp-like proteins. It was previously shown that the pore components of this oligopeptide permease, OppBCDF, were required for the import of PapR. However, the role of OppA, the substrate-binding protein (SBP), was not investigated. Here, we demonstrated that OppA is not the only SBP involved in the recognition of PapR, and that several other OppA-like proteins can allow the import of this peptide. Altogether, these data complete our model of quorum sensing during the lifecycle of Bt and indicate that RNPPs integrate environmental conditions, as well as cell density, to coordinate the behavior of the bacteria throughout growth. PMID:26779156

  20. CodY Regulates the Activity of the Virulence Quorum Sensor PlcR by Controlling the Import of the Signaling Peptide PapR in Bacillus thuringiensis.

    PubMed

    Slamti, Leyla; Lemy, Christelle; Henry, Céline; Guillot, Alain; Huillet, Eugénie; Lereclus, Didier

    2015-01-01

    In Gram-positive bacteria, cell-cell communication mainly relies on cytoplasmic sensors of the RNPP family. Activity of these regulators depends on their binding to secreted signaling peptides that are imported into the cell. These quorum sensing regulators control important biological functions in bacteria of the Bacillus cereus group, such as virulence and necrotrophism. The RNPP quorum sensor PlcR, in complex with its cognate signaling peptide PapR, is the main regulator of virulence in B. cereus and Bacillus thuringiensis (Bt). Recent reports have shown that the global stationary phase regulator CodY, involved in adaptation to nutritional limitation, is required for the expression of virulence genes belonging to the PlcR regulon. However, the mechanism underlying this regulation was not described. Using genetics and proteomics approaches, we showed that CodY regulates the expression of the virulence genes through the import of PapR. We report that CodY positively controls the production of the proteins that compose the oligopeptide permease OppABCDF, and of several other Opp-like proteins. It was previously shown that the pore components of this oligopeptide permease, OppBCDF, were required for the import of PapR. However, the role of OppA, the substrate-binding protein (SBP), was not investigated. Here, we demonstrated that OppA is not the only SBP involved in the recognition of PapR, and that several other OppA-like proteins can allow the import of this peptide. Altogether, these data complete our model of quorum sensing during the lifecycle of Bt and indicate that RNPPs integrate environmental conditions, as well as cell density, to coordinate the behavior of the bacteria throughout growth. PMID:26779156

  1. Temperature responses of the Rubisco maximum carboxylase activity across domains of life: phylogenetic signals, trade-offs, and importance for carbon gain.

    PubMed

    Galmés, J; Kapralov, M V; Copolovici, L O; Hermida-Carrera, C; Niinemets, Ü

    2015-02-01

    Temperature response of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) catalytic properties directly determines the CO2 assimilation capacity of photosynthetic organisms as well as their survival in environments with different thermal conditions. Despite unquestionable importance of Rubisco, the comprehensive analysis summarizing temperature responses of Rubisco traits across lineages of carbon-fixing organisms is lacking. Here, we present a review of the temperature responses of Rubisco carboxylase specific activity (c(cat)(c)) within and across domains of life. In particular, we consider the variability of temperature responses, and their ecological, physiological, and evolutionary controls. We observed over two-fold differences in the energy of activation (ΔH(a)) among different groups of photosynthetic organisms, and found significant differences between C3 plants from cool habitats, C3 plants from warm habitats and C4 plants. According to phylogenetically independent contrast analysis, ΔH(a) was not related to the species optimum growth temperature (T growth), but was positively correlated with Rubisco specificity factor (S(c/o)) across all organisms. However, when only land plants were analyzed, ΔH(a) was positively correlated with both T(growth) and S(c/o), indicating different trends for these traits in plants versus unicellular aquatic organisms, such as algae and bacteria. The optimum temperature (T(opt)) for k(cat)(c) correlated with S(c/o) for land plants and for all organisms pooled, but the effect of T growth on T(opt) was driven by species phylogeny. The overall phylogenetic signal was significant for all analyzed parameters, stressing the importance of considering the evolutionary framework and accounting for shared ancestry when deciphering relationships between Rubisco kinetic parameters. We argue that these findings have important implications for improving global photosynthesis models.

  2. Identification of Functionally Important Residues of the Silkmoth Pheromone Biosynthesis-activating Neuropeptide Receptor, an Insect Ortholog of the Vertebrate Neuromedin U Receptor*

    PubMed Central

    Kawai, Takeshi; Katayama, Yukie; Guo, Linjun; Liu, Desheng; Suzuki, Tatsuya; Hayakawa, Kou; Lee, Jae Min; Nagamine, Toshihiro; Hull, J. Joe; Matsumoto, Shogo; Nagasawa, Hiromichi; Tanokura, Masaru; Nagata, Koji

    2014-01-01

    The biosynthesis of sex pheromone components in many lepidopteran insects is regulated by the interaction between pheromone biosynthesis-activating neuropeptide (PBAN) and the PBAN receptor (PBANR), a class A G-protein-coupled receptor. To identify functionally important amino acid residues in the silkmoth PBANR, a series of 27 alanine substitutions was generated using a PBANR chimera C-terminally fused with enhanced GFP. The PBANR mutants were expressed in Sf9 insect cells, and their ability to bind and be activated by a core PBAN fragment (C10PBANR2K) was monitored. Among the 27 mutants, 23 localized to the cell surface of transfected Sf9 cells, whereas the other four remained intracellular. Reduced binding relative to wild type was observed with 17 mutants, and decreased Ca2+ mobilization responses were observed with 12 mutants. Ala substitution of Glu-95, Glu-120, Asn-124, Val-195, Phe-276, Trp-280, Phe-283, Arg-287, Tyr-307, Thr-311, and Phe-319 affected both binding and Ca2+ mobilization. The most pronounced effects were observed with the E120A mutation. A molecular model of PBANR indicated that the functionally important PBANR residues map to the 2nd, 3rd, 6th, and 7th transmembrane helices, implying that the same general region of class A G-protein-coupled receptors recognizes both peptidic and nonpeptidic ligands. Docking simulations suggest similar ligand-receptor recognition interactions for PBAN-PBANR and the orthologous vertebrate pair, neuromedin U (NMU) and NMU receptor (NMUR). The simulations highlight the importance of two glutamate residues, Glu-95 and Glu-120, in silkmoth PBANR and Glu-117 and Glu-142 in human NMUR1, in the recognition of the most functionally critical region of the ligands, the C-terminal residue and amide. PMID:24847080

  3. Proteasome activity is important for replication recovery, CHK1 phosphorylation and prevention of G2 arrest after low-dose formaldehyde.

    PubMed

    Ortega-Atienza, Sara; Green, Samantha E; Zhitkovich, Anatoly

    2015-07-15

    Formaldehyde (FA) is a human carcinogen with numerous sources of environmental and occupational exposures. This reactive aldehyde is also produced endogenously during metabolism of drugs and other processes. DNA-protein crosslinks (DPCs) are considered to be the main genotoxic lesions for FA. Accumulating evidence suggests that DPC repair in high eukaryotes involves proteolysis of crosslinked proteins. Here, we examined a role of the main cellular proteolytic machinery proteasomes in toxic responses of human lung cells to low FA doses. We found that transient inhibition of proteasome activity increased cytotoxicity and diminished clonogenic viability of FA-treated cells. Proteasome inactivation exacerbated suppressive effects of FA on DNA replication and increased the levels of the genotoxic stress marker γ-H2AX in normal human cells. A transient loss of proteasome activity in FA-exposed cells also caused delayed perturbations of cell cycle, which included G2 arrest and a depletion of S-phase populations at FA doses that had no effects in control cells. Proteasome activity diminished p53-Ser15 phosphorylation but was important for FA-induced CHK1 phosphorylation, which is a biochemical marker of DPC proteolysis in replicating cells. Unlike FA, proteasome inhibition had no effect on cell survival and CHK1 phosphorylation by the non-DPC replication stressor hydroxyurea. Overall, we obtained evidence for the importance of proteasomes in protection of human cells against biologically relevant doses of FA. Biochemically, our findings indicate the involvement of proteasomes in proteolytic repair of DPC, which removes replication blockage by these highly bulky lesions.

  4. Importance of sports during youth and exercise barriers in 20- to 29-year-old male nonathletes differently motivated for regular physical activities.

    PubMed

    Leyk, Dieter; Witzki, Alexander; Sievert, Alexander; Rohde, Ulrich; Moedl, Anne; Rüther, Thomas; Löllgen, Herbert; Hackfort, Dieter

    2012-07-01

    The number of sedentary young adults has dramatically increased in past decades, and sedentary lifestyles are adopted at an increasingly earlier age. Little is known about barriers or predictors to (re)initiate regular physical activity in this group. The purpose of the study is to (a) identify subgroups in nonathletes differing in their amenability to physical exercise, (b) to analyze them for differences in barriers and intention to exercise, and (c) compare importance of sports during youth in nonathletes to trained peers. Using a health and fitness questionnaire 589 nonathletes were queried in the cross-sectional survey and compared with 270 trained peers. Athletic abstainers (A), lower (L), and higher (H) motivated nonathletes were separated based on previous engagement in sports. Of the nonathletes, 54.7% reported only 1 barrier to exercise. Although this feature was most prominent in H (71.5%), the other groups showed significantly more barriers and a broader distribution. Similar characteristics but minor differences were observed for perceived importance of sports during youth. The most significant differences between athletes and nonathletes emerged enquiring the attitude and activity of the parents. The majority of nonathletes (72.8%) indicate their intention to exercise in the future. Their intention differed significantly in H (88.1%), L (76.1%), and A (59.1%). However, there are good reasons to doubt that most of those intending nonathletes will actually become physically active. Even in the analyzed narrow age range of men different motivated groups of nonathletes were found. Because of the differences in receptiveness and approachability health promotion policies may not only consider the often recommended tailored interventions but also carefully designed incentive programs.

  5. Overweight and Obese Adolescent Girls: The Importance of Promoting Sensible Eating and Activity Behaviors from the Start of the Adolescent Period

    PubMed Central

    Todd, Alwyn S.; Street, Steven J.; Ziviani, Jenny; Byrne, Nuala M.; Hills, Andrew P.

    2015-01-01

    The adolescent period is associated with changes in eating and activity behaviors in girls. Less reliance on parental provision and choice of food, coupled with a decrease in participation in physical activity and sport, can create an energy imbalance, predisposing to weight gain. Physiological alterations to body composition, reduction in insulin sensitivity, and psychological adjustments may further amplify the risk of becoming overweight and maintaining an unhealthy level of body fat into childbearing years. During pregnancy excess body fat is a risk factor for poor pregnancy outcomes and may predispose an infant to a lifelong heightened risk of being overweight and developing chronic disease. Interventions aimed at preventing the accumulation of body fat in adolescent girls and young women may have far reaching impact and be critically important in reducing intergenerational weight gain. Lifestyle interventions in adolescence have the potential to modify adult obesity risk by switching at-risk individuals from a high to lower obesity risk trajectory. This paper discusses multiple approaches to assist at-risk individuals reduce obesity risk. A key focus is placed on engagement in food preparation and choice, and opportunities for physical activity and sport. Support, education, and opportunity at home and at school, are often associated with the success of lifestyle interventions, and may enable adolescents to make positive choices, and engage in health promoting behaviors during adolescence and childbearing years. PMID:25690003

  6. Regional mutagenesis of the gene encoding the phage Mu late gene activator C identifies two separate regions important for DNA binding

    PubMed Central

    Jiang, Yide; Howe, Martha M.

    2008-01-01

    Lytic development of bacteriophage Mu is controlled by a regulatory cascade and involves three phases of transcription: early, middle and late. Late transcription requires the host RNA polymerase holoenzyme and a 16.5-kDa Mu-encoded activator protein C. Consistent with these requirements, the four late promoters Plys, PI, PP and Pmom have recognizable −10 hexamers but lack typical −35 hexamers. The C protein binds to a 16-bp imperfect dyad-symmetrical sequence element centered at −43.5 and overlapping the −35 region. Based on the crystal structure of the closely related Mor protein, the activator of Mu middle transcription, we predict that two regions of C are involved in DNA binding: a helix-turn-helix region and a β-strand region linking the dimerization and helix-turn-helix domains. To test this hypothesis, we carried out mutagenesis of the corresponding regions of the C gene by degenerate oligonucleotide-directed PCR and screened the resulting mutants for their ability to activate a Plys-galK fusion. Analysis of the mutant proteins by gel mobility shift, β-galactosidase and polyacrylamide gel electrophoresis assays identified a number of amino acid residues important for C DNA binding in both regions. PMID:18838393

  7. Overweight and obese adolescent girls: the importance of promoting sensible eating and activity behaviors from the start of the adolescent period.

    PubMed

    Todd, Alwyn S; Street, Steven J; Ziviani, Jenny; Byrne, Nuala M; Hills, Andrew P

    2015-02-01

    The adolescent period is associated with changes in eating and activity behaviors in girls. Less reliance on parental provision and choice of food, coupled with a decrease in participation in physical activity and sport, can create an energy imbalance, predisposing to weight gain. Physiological alterations to body composition, reduction in insulin sensitivity, and psychological adjustments may further amplify the risk of becoming overweight and maintaining an unhealthy level of body fat into childbearing years. During pregnancy excess body fat is a risk factor for poor pregnancy outcomes and may predispose an infant to a lifelong heightened risk of being overweight and developing chronic disease. Interventions aimed at preventing the accumulation of body fat in adolescent girls and young women may have far reaching impact and be critically important in reducing intergenerational weight gain. Lifestyle interventions in adolescence have the potential to modify adult obesity risk by switching at-risk individuals from a high to lower obesity risk trajectory. This paper discusses multiple approaches to assist at-risk individuals reduce obesity risk. A key focus is placed on engagement in food preparation and choice, and opportunities for physical activity and sport. Support, education, and opportunity at home and at school, are often associated with the success of lifestyle interventions, and may enable adolescents to make positive choices, and engage in health promoting behaviors during adolescence and childbearing years. PMID:25690003

  8. Activation of the Nrf2 Cell Defense Pathway by Ancient Foods: Disease Prevention by Important Molecules and Microbes Lost from the Modern Western Diet.

    PubMed

    Senger, Donald R; Li, Dan; Jaminet, Shou-Ching; Cao, Shugeng

    2016-01-01

    The Nrf2 (NFE2L2) cell defense pathway protects against oxidative stress and disorders including cancer and neurodegeneration. Although activated modestly by oxidative stress alone, robust activation of the Nrf2 defense mechanism requires the additional presence of co-factors that facilitate electron exchange. Various molecules exhibit this co-factor function, including sulforaphane from cruciferous vegetables. However, natural co-factors that are potent and widely available from dietary sources have not been identified previously. The objectives of this study were to investigate support of the Nrf2 cell defense pathway by the alkyl catechols: 4-methylcatechol, 4-vinylcatechol, and 4-ethylcatechol. These small electrochemicals are naturally available from numerous sources but have not received attention. Findings reported here illustrate that these compounds are indeed potent co-factors for activation of the Nrf2 pathway both in vitro and in vivo. Each strongly supports expression of Nrf2 target genes in a variety of human cell types; and, in addition, 4-ethylcatechol is orally active in mice. Furthermore, findings reported here identify important and previously unrecognized sources of these compounds, arising from biotransformation of common plant compounds by lactobacilli that express phenolic acid decarboxylase. Thus, for example, Lactobacillus plantarum, Lactobacillus brevis, and Lactobacillus collinoides, which are consumed from a diet rich in traditionally fermented foods and beverages, convert common phenolic acids found in fruits and vegetables to 4-vinylcatechol and/or 4-ethylcatechol. In addition, all of the alkyl catechols are found in wood smoke that was used widely for food preservation. Thus, the potentially numerous sources of alkyl catechols in traditional foods suggest that these co-factors were common in ancient diets. However, with radical changes in food preservation, alkyl catechols have been lost from modern foods. The absence of alkyl

  9. Activation of the Nrf2 Cell Defense Pathway by Ancient Foods: Disease Prevention by Important Molecules and Microbes Lost from the Modern Western Diet.

    PubMed

    Senger, Donald R; Li, Dan; Jaminet, Shou-Ching; Cao, Shugeng

    2016-01-01

    The Nrf2 (NFE2L2) cell defense pathway protects against oxidative stress and disorders including cancer and neurodegeneration. Although activated modestly by oxidative stress alone, robust activation of the Nrf2 defense mechanism requires the additional presence of co-factors that facilitate electron exchange. Various molecules exhibit this co-factor function, including sulforaphane from cruciferous vegetables. However, natural co-factors that are potent and widely available from dietary sources have not been identified previously. The objectives of this study were to investigate support of the Nrf2 cell defense pathway by the alkyl catechols: 4-methylcatechol, 4-vinylcatechol, and 4-ethylcatechol. These small electrochemicals are naturally available from numerous sources but have not received attention. Findings reported here illustrate that these compounds are indeed potent co-factors for activation of the Nrf2 pathway both in vitro and in vivo. Each strongly supports expression of Nrf2 target genes in a variety of human cell types; and, in addition, 4-ethylcatechol is orally active in mice. Furthermore, findings reported here identify important and previously unrecognized sources of these compounds, arising from biotransformation of common plant compounds by lactobacilli that express phenolic acid decarboxylase. Thus, for example, Lactobacillus plantarum, Lactobacillus brevis, and Lactobacillus collinoides, which are consumed from a diet rich in traditionally fermented foods and beverages, convert common phenolic acids found in fruits and vegetables to 4-vinylcatechol and/or 4-ethylcatechol. In addition, all of the alkyl catechols are found in wood smoke that was used widely for food preservation. Thus, the potentially numerous sources of alkyl catechols in traditional foods suggest that these co-factors were common in ancient diets. However, with radical changes in food preservation, alkyl catechols have been lost from modern foods. The absence of alkyl

  10. Activation of the Nrf2 Cell Defense Pathway by Ancient Foods: Disease Prevention by Important Molecules and Microbes Lost from the Modern Western Diet

    PubMed Central

    Senger, Donald R.; Li, Dan; Jaminet, Shou-Ching; Cao, Shugeng

    2016-01-01

    The Nrf2 (NFE2L2) cell defense pathway protects against oxidative stress and disorders including cancer and neurodegeneration. Although activated modestly by oxidative stress alone, robust activation of the Nrf2 defense mechanism requires the additional presence of co-factors that facilitate electron exchange. Various molecules exhibit this co-factor function, including sulforaphane from cruciferous vegetables. However, natural co-factors that are potent and widely available from dietary sources have not been identified previously. The objectives of this study were to investigate support of the Nrf2 cell defense pathway by the alkyl catechols: 4-methylcatechol, 4-vinylcatechol, and 4-ethylcatechol. These small electrochemicals are naturally available from numerous sources but have not received attention. Findings reported here illustrate that these compounds are indeed potent co-factors for activation of the Nrf2 pathway both in vitro and in vivo. Each strongly supports expression of Nrf2 target genes in a variety of human cell types; and, in addition, 4-ethylcatechol is orally active in mice. Furthermore, findings reported here identify important and previously unrecognized sources of these compounds, arising from biotransformation of common plant compounds by lactobacilli that express phenolic acid decarboxylase. Thus, for example, Lactobacillus plantarum, Lactobacillus brevis, and Lactobacillus collinoides, which are consumed from a diet rich in traditionally fermented foods and beverages, convert common phenolic acids found in fruits and vegetables to 4-vinylcatechol and/or 4-ethylcatechol. In addition, all of the alkyl catechols are found in wood smoke that was used widely for food preservation. Thus, the potentially numerous sources of alkyl catechols in traditional foods suggest that these co-factors were common in ancient diets. However, with radical changes in food preservation, alkyl catechols have been lost from modern foods. The absence of alkyl

  11. Mnk1 and 2 are dispensable for T-cell development and activation but important for the pathogenesis of experimental autoimmune encephalitis

    PubMed Central

    Gorentla, Balachandra K; Krishna, Sruti; Shin, Jinwook; Inoue, Makoto; Shinohara, Mari L.; Grayson, Jason M.; Fukunaga, Rikiro; Zhong, Xiao-Ping

    2012-01-01

    T-cell development and activation are usually accompanied by expansion and production of numerous proteins that require active translation. The eukaryotic translation initiation factor 4E (eIF4E) binds to the 5' cap structure of mRNA and is critical for cap-dependent translational initiation. It has been hypothesized that MAPK-interacting kinase 1 and 2 (Mnk1/2) promote cap-dependent translation by phosphorylating eIF4E at serine 209 (S209). Pharmacological studies utilizing inhibitors have suggested that Mnk1/2 play important roles in T-cells. However, genetic evidence supporting such conclusions is lacking. Moreover, the signaling pathways that regulate Mnk1/2 in T-cells remain unclear. We demonstrated here that T-cell receptor (TCR) engagement activates Mnk1/2 in primary T-cells. Such activation is dependent on Ras-Erk1/2 signaling and is inhibited by diacylglycerol kinases α and ζ. Mnk1/2 double deficiency in mice abolishes TCR-induced eIF4E S209 phosphorylation, indicating their absolute requirement for eIF4E S209 phosphorylation. However, Mnk1/2 double deficiency does not affect the development of conventional αβ T-cells, regulatory T-cells, or NKT-cells. Furthermore, T-cell activation, in vivo primary and memory CD8 T-cell responses to microbial infection, and NKT-cell cytokine production were not obviously altered by Mnk1/2 deficiency. Although Mnk1/2 deficiency causes decreased IL-17 and IFNγ production by CD4 T-cells following immunization of mice with myelin oligodendrocyte glycoprotein peptide in complete Freud's adjuvant, correlating with milder experimental autoimmune encephalitis scores, it does not affect T helper cell differentiation in vitro. Together, these data suggest that Mnk1/2 play a minimal role in T-cell development and activation but may regulate non-T-cell lineages to control Th1/Th17 differentiation in vivo. PMID:23269249

  12. Energy imparted-based estimates of the effect of z overscanning on adult and pediatric patient effective doses from multi-slice computed tomography

    SciTech Connect

    Theocharopoulos, Nicholas; Damilakis, John; Perisinakis, Kostas; Gourtsoyiannis, Nicholas

    2007-04-15

    In the present study effective dose values normalized to computed tomography dose index measured free in air were calculated for adult, newborn, 1, 5, 10 and 15 year old patients regarding scans of the head, chest, abdomen, pelvis, abdomen and pelvis, and trunk, using the energy imparted method. The effect of z overscanning on patient doses was accounted for, and normalized doses are provided for varying beam collimation, pitch and reconstruction slice width values. The contribution of overscanning depends on patient age, anatomic region imaged, acquisition and reconstruction settings. For a head scan it constitutes 15% of the adult effective dose and 24% of the effective dose to a newborn but for an abdomen scan it may be as high as 58% for a newborn and 31% for an adult. The ratios of normalized pediatric doses relative to that for adults for helical scans depend not only on age but also on acquisition and reconstruction parameters, because of variations in the relative distance between the primary beam and the radiosensitive tissues/organs of the body. Regarding scans of the trunk, pediatric doses are up to a factor of 2.5 times higher compared to adult doses (abdominal scans), whereas for scans of the head up to a factor of 1.5. Increasing the pitch value of helical scans while maintaining the same effective mAs setting, and hence noise levels, leads to an increase in patient doses which depends on age, body region, scan and reconstruction parameters. The % difference between doses at pitch 1.5 and pitch 1 is more pronounced in the abdominal region (14% increase for adults) and in young patients (31% in a newborn and 18% in a 10 year old patient) and it is minimal in head scans (4% increase in newborns and 1% in adults). If multiple body regions are to be imaged, doses to adults can be reduced by up to 15% and 36% to children by performing single long-range scans. Scanning adult patients at 100 kVp instead of 120 kVp, results in a 32% reduction in effective

  13. Sensitivity of coefficients for converting entrance surface dose and kerma-area product to effective dose and energy imparted to the patient

    NASA Astrophysics Data System (ADS)

    Wise, K. N.; Sandborg, M.; Persliden, J.; Alm Carlsson, G.

    1999-08-01

    We investigate the sensitivity of the conversions from entrance surface dose (ESD) or kerma-area product (KAP) to effective dose (E) or to energy imparted to the patient (varepsilon) to the likely variations in tube potential, field size, patient size and sex which occur in clinical work. As part of a factorial design study for chest and lumbar spine examinations, the tube potentials were varied to be ±10% of the typical values for the examinations while field sizes and the positions of the field centres were varied to be representative of values drawn from measurements on patient images. Variation over sex and patient size was based on anthropomorphic phantoms representing males and females of ages 15 years (small adult) and 21 years (reference adult). All the conversion coefficients were estimated using a mathematical phantom programmed with the Monte Carlo code EGS4 for all factor combinations and analysed statistically to derive factor effects. In general, the factors studied behaved independently in the sense that interaction of the physical factors generally gave no more than a 5% variation in a conversion coefficient. Taken together, variation of patient size, sex, field size and field position can lead to significant variation of E/KAP by up to a factor of 2, of E/ESD by up to a factor of 3, of varepsilon/KAP by a factor of 1.3 and of varepsilon/ESD by up to a factor of 2. While KAP is preferred to determine varepsilon, the results show no strong preference of KAP over ESD in determining E. The mean absorbed dose (barD) in the patient obtained by dividing

  14. Insecticidal, fumigant, and repellent activities of sweet wormwood oil and its individual components against red imported fire ant workers (Hymenoptera: Formicidae).

    PubMed

    Zhang, Ning; Tang, Liang; Hu, Wei; Wang, Kun; Zhou, You; Li, Hong; Huang, Congling; Chun, Jiong; Zhang, Zhixiang

    2014-01-01

    In total, 29 compounds from sweet wormwood (Artemisia annua L.) oil were identified using gas chromatography-mass spectrometry. The five active components were D-camphor, linalool, cineole, α-terpineol, and L(-)-borneol. The effectiveness of A. annua oil, as well as d-camphor, linalool, cineole, α-terpineol, and L(-)-borneol, as fumigants, contact insecticides, and repellents, were tested on the red imported fire ant Solenopsis invicta Buren. The results indicated that A. annua oil has no significant topical toxicity; however, the spray contact test revealed that it has strong insecticidal activity and the inhibitory effect is stronger during closed exposure than during open exposure. In the fumigant test, cineole and D-camphor exhibited strong fumigant toxicity on minor and major S. invicta workers. They also caused 100% mortality at 5, 3, 2, and 1 mg/centrifuge tube but not at 0.5 mg/centrifuge tube. The mortality rates of linalool, α-terpineol, and L(-)-borneol exceeded 80% at 5, 3, and 2 mg/centrifuge tube. In the repellent test, cineole and d-camphor showed significant repellency at 100, 10, and 1 mg/kg. However, linalool, α-terpineol, and L(-)-borneol significantly facilitated digging at 10 and 1 mg/kg.

  15. JAIBA, a class-II HD-ZIP transcription factor involved in the regulation of meristematic activity, and important for correct gynoecium and fruit development in Arabidopsis.

    PubMed

    Zúñiga-Mayo, Victor M; Marsch-Martínez, Nayelli; de Folter, Stefan

    2012-07-01

    The gynoecium is one of the most complex organs of a plant. After fertilization, it becomes a fruit, which has two important functions: to protect the seeds while they develop and to disperse them at maturity. The development and patterning of the gynoecium and later fruit must be finely regulated to ensure the survival of the species that produces them. The process that leads to successful fruit formation starts at early stages of floral meristem development and follows a series of chronologically successive events. In this work we report the functional characterization of the class-II homeodomain leucine zipper (HD-ZIP) JAIBA (JAB) gene. Mutant jab plants show sporophytic defects in male and female reproductive development, and combined with crabs claw cause defects in the floral meristem (FM) determination process and gynoecium medial tissue development. This suggests that proper FM determination is required for gynoecium medial tissue development, and indicates that JAB and CRC are necessary for both processes. Furthermore, the JAB protein interacts with transcription factors known to regulate meristematic activity, fruit development, and floral meristem determinacy. The sporophytic effect on pollen and embryo sac development might be an independent and later function of JAB. In summary, we present data that indicates that the JAB gene regulates meristematic activity in different tissues, and that it is necessary for the correct formation of the gynoecium at different stages, contributing to a crucial process in plant life: proper fruit development. PMID:22409594

  16. Insecticidal, fumigant, and repellent activities of sweet wormwood oil and its individual components against red imported fire ant workers (Hymenoptera: Formicidae).

    PubMed

    Zhang, Ning; Tang, Liang; Hu, Wei; Wang, Kun; Zhou, You; Li, Hong; Huang, Congling; Chun, Jiong; Zhang, Zhixiang

    2014-01-01

    In total, 29 compounds from sweet wormwood (Artemisia annua L.) oil were identified using gas chromatography-mass spectrometry. The five active components were D-camphor, linalool, cineole, α-terpineol, and L(-)-borneol. The effectiveness of A. annua oil, as well as d-camphor, linalool, cineole, α-terpineol, and L(-)-borneol, as fumigants, contact insecticides, and repellents, were tested on the red imported fire ant Solenopsis invicta Buren. The results indicated that A. annua oil has no significant topical toxicity; however, the spray contact test revealed that it has strong insecticidal activity and the inhibitory effect is stronger during closed exposure than during open exposure. In the fumigant test, cineole and D-camphor exhibited strong fumigant toxicity on minor and major S. invicta workers. They also caused 100% mortality at 5, 3, 2, and 1 mg/centrifuge tube but not at 0.5 mg/centrifuge tube. The mortality rates of linalool, α-terpineol, and L(-)-borneol exceeded 80% at 5, 3, and 2 mg/centrifuge tube. In the repellent test, cineole and d-camphor showed significant repellency at 100, 10, and 1 mg/kg. However, linalool, α-terpineol, and L(-)-borneol significantly facilitated digging at 10 and 1 mg/kg. PMID:25525106

  17. SARS-coronavirus spike S2 domain flanked by cysteine residues C822 and C833 is important for activation of membrane fusion.

    PubMed

    Madu, Ikenna G; Belouzard, Sandrine; Whittaker, Gary R

    2009-10-25

    The S2 domain of the coronavirus spike (S) protein is known to be responsible for mediating membrane fusion. In addition to a well-recognized cleavage site at the S1-S2 boundary, a second proteolytic cleavage site has been identified in the severe acute respiratory syndrome coronavirus (SARS-CoV) S2 domain (R797). C-terminal to this S2 cleavage site is a conserved region flanked by cysteine residues C822 and C833. Here, we investigated the importance of this well conserved region for SARS-CoV S-mediated fusion activation. We show that the residues between C822-C833 are well conserved across all coronaviruses. Mutagenic analysis of SARS-CoV S, combined with cell-cell fusion and pseudotyped virion infectivity assays, showed a critical role for the core-conserved residues C822, D830, L831, and C833. Based on available predictive models, we propose that the conserved domain flanked by cysteines 822 and 833 forms a loop structure that interacts with components of the SARS-CoV S trimer to control the activation of membrane fusion.

  18. SARS-coronavirus spike S2 domain flanked by cysteine residues C822 and C833 is important for activation of membrane fusion

    PubMed Central

    Madu, Ikenna G.; Belouzard, Sandrine; Whittaker, Gary R.

    2013-01-01

    The S2 domain of the coronavirus spike (S) protein is known to be responsible for mediating membrane fusion. In addition to a well-recognized cleavage site at the S1–S2 boundary, a second proteolytic cleavage site has been identified in the severe acute respiratory syndrome coronavirus (SARS-CoV) S2 domain (R797). C terminal to this S2 cleavage site is a conserved region flanked by cysteine residues C822 and C833. Here, we investigated the importance of this well conserved region for SARS-CoV S-mediated fusion activation. We show that the residues between C822-C833 are well conserved across all coronaviruses. Mutagenic analysis of SARS-CoV S, combined with cell–cell fusion and pseudotyped virion infectivity assays, showed a critical role for the core-conserved residues C822, D830, L831, and C833. Based on available predictive models, we propose that the conserved domain flanked by cysteines 822 and 833 forms a loop structure that interacts with components of the SARS-CoV S trimer to control the activation of membrane fusion. PMID:19717178

  19. SARS-coronavirus spike S2 domain flanked by cysteine residues C822 and C833 is important for activation of membrane fusion

    SciTech Connect

    Madu, Ikenna G.; Belouzard, Sandrine; Whittaker, Gary R.

    2009-10-25

    The S2 domain of the coronavirus spike (S) protein is known to be responsible for mediating membrane fusion. In addition to a well-recognized cleavage site at the S1-S2 boundary, a second proteolytic cleavage site has been identified in the severe acute respiratory syndrome coronavirus (SARS-CoV) S2 domain (R797). C-terminal to this S2 cleavage site is a conserved region flanked by cysteine residues C822 and C833. Here, we investigated the importance of this well conserved region for SARS-CoV S-mediated fusion activation. We show that the residues between C822-C833 are well conserved across all coronaviruses. Mutagenic analysis of SARS-CoV S, combined with cell-cell fusion and pseudotyped virion infectivity assays, showed a critical role for the core-conserved residues C822, D830, L831, and C833. Based on available predictive models, we propose that the conserved domain flanked by cysteines 822 and 833 forms a loop structure that interacts with components of the SARS-CoV S trimer to control the activation of membrane fusion.

  20. Activity of the Na,K-ATPase alpha4 isoform is important for membrane potential, intracellular Ca2+, and pH to maintain motility in rat spermatozoa.

    PubMed

    Jimenez, Tamara; Sánchez, Gladis; Wertheimer, Eva; Blanco, Gustavo

    2010-05-01

    While the function of the ubiquitous Na,K-ATPase alpha1 subunit has been well documented, the role of the sperm-specific alpha4 isoform of this ion transporter is less known. We have explored the importance of alpha4 in rat sperm physiology by taking advantage of the high sensitivity of this isoform for the inhibitor ouabain. Using concentrations that selectively block alpha4 activity, we found ouabain to reduce not only sperm total motility, but also multiple parameters of sperm movement, including progressive motility, straight line, curvilinear, and average path velocities, lateral head displacement, beat cross frequency, and linearity. According to a direct role of alpha4 in Na(+) transport, ouabain inhibition of alpha4 increased [Na(+)](i) in the male gametes. In addition, interference of alpha4 activity with ouabain produced cell membrane depolarization, diminished pH, and increased [Ca(2)(+)](i) in spermatozoa. Inhibition of alpha4 was sufficient to cause all these effects and additional blockage of alpha1, the other Na,K-ATPase alpha isoform expressed in sperm, and higher doses of ouabain did not result in further changes in the cell parameters studied. These results show that alpha4 is the Na,K-ATPase isoform primarily involved in controlling the transmembrane Na(+) gradient in sperm, and that alpha4 activity is necessary for maintaining membrane potential, [Ca(2)(+)](i), and [H(+)](i) in the cells. The high dependence of sperm motility on membrane excitability, [Ca(2)(+)](i), and acid-base balance suggests that their regulation is the mechanism by which alpha4 maintains motility of the male gametes.

  1. Mechanism of the reaction catalyzed by acetoacetate decarboxylase. Importance of lysine 116 in determining the pKa of active-site lysine 115.

    PubMed

    Highbarger, L A; Gerlt, J A; Kenyon, G L

    1996-01-01

    Acetoacetate decarboxylase from Clostridium acetobutylicum (AAD) catalyzes the decarboxylation of acetoacetate via a Schiff base intermediate [Hamilton, G. A., & Westheimer, F. H. (1959) J. Am. Chem. Soc. 81, 6332; Fridovich, I., & Westheimer F. H. (1962) J. Am. Chem. Soc. 84, 3208]. The pKa of the active-site lysine (Lys 115) is 6.0, 4.5 pKa units less than the pKa of lysine in solution [Kokesh, F. C., & Westheimer, F. H. (1971) J. Am. Chem. Soc. 93, 7270; Frey, P. A., Kokesh, F. C., & Westheimer, F. H. (1971) J. Am. Chem. Soc. 93, 7266; Schmidt, D. E., Jr., & Westheimer, F. H. (1971) Biochemistry 10, 1249]. Westheimer and co-workers hypothesized that the pKa of Lys 115 is decreased by its spatial proximity to the epsilon-ammonium group of Lys 116. We have investigated this proposal by studying site-directed mutants of Lys 115 and Lys 116. Two substitutions for Lys 115 (K115C and K115Q) were both catalytically inactive at pH 5.95, the pH optimum of wild type AAD, demonstrating the importance of this residue in catalysis. Activity could be restored to K115C by aminoethylation with 2-bromoethyl-ammonium bromide (2-BEAB). Substitutions for Lys 116 (K116C, K116N, and K116R) had reduced but significant activities at pH 5.95. The effects of Lys 116 on the pKa of Lys 115 in the mutant AADs were evaluated following imine formation with 5-nitrosalicylaldehyde and reduction with NaBH4. Whereas the pKa of Lys 115 in K116R is similar to that observed for wild type AAD, the pKaS of Lys 115 in K116C and K116N were elevated to > 9.2. Alkylation of Cys 116 in K116C with 2-BEAB resulted in both significant activation and restoration of the pKa of Lys 115 to 5.9. These data support Westheimer's hypothesis that the pKa of the Schiff base-forming Lys 115 is decreased by its spatial proximity to the epsilon-ammonium group of Lys 116. PMID:8555196

  2. The importance of earthquake-induced landslides to long-term slope erosion and slope-failure hazards in seismically active regions

    USGS Publications Warehouse

    Keefer, D.K.

    1994-01-01

    This paper describes a general method for determining the amount of earthquake-induced landsliding that occurs in a seismically active region over time; this determination can be used as a quantitative measure of the long-term hazard from seismically triggered landslides as well as a measure of the importance of this process to regional slope-erosion rates and landscape evolution. The method uses data from historical earthquakes to relate total volume of landslide material dislodged by an earthquake to the magnitude, M, and seismic moment, M0, of the earthquake. From worldwide data, a linear-regression relation between landslide volume, V, and M0 is determined as: V = M0/1018.9(?? 0.13), where V is measured in m3 and M0 is in dyn-cm. To determine the amount of earthquake-generated landsliding over time, this relation is combined with data on seismic-moment release for a particular region, which may be derived from either earthquake-history or fault-slip data. The form of the M0-V relation allows the rate of production of earthquake-induced landslides over time to be determined from total rate of seismic-moment release without regard to the distribution of individual events, thus simplifying and generalizing the determination. Application of the method to twelve seismically active regions, with areas ranging from 13,275 to 2,308,000 km2, shows that erosion rates from earthquake-induced landslides vary significantly from region to region. Of the regions studied, the highest rates were determined for the island of Hawaii, New Zealand, western New Guinea, and the San Francisco Bay region of California. Significantly lower rates were determined for Iran, Tibet, the Sierra Nevada-Great Basin region of California, and central Japan (for the time period from 715 AD to the present). Intermediate rates were determined for Peru, southern California, onshore California, Turkey, and central Japan (for the time period from 1586 AD to the present). To determine the relative, long

  3. Key importance of small RNA binding for the activity of a glycine-tryptophan (GW) motif-containing viral suppressor of RNA silencing.

    PubMed

    Pérez-Cañamás, Miryam; Hernández, Carmen

    2015-01-30

    Viruses express viral suppressors of RNA silencing (VSRs) to counteract RNA silencing-based host defenses. Although virtually all stages of the antiviral silencing pathway can be inhibited by VSRs, small RNAs (sRNAs) and Argonaute (AGO) proteins seem to be the most frequent targets. Recently, GW/WG motifs of some VSRs have been proposed to dictate their suppressor function by mediating interaction with AGO(s). Here we have studied the VSR encoded by Pelargonium line pattern virus (family Tombusviridae). The results show that p37, the viral coat protein, blocks RNA silencing. Site-directed mutagenesis of some p37 sequence traits, including a conserved GW motif, allowed generation of suppressor-competent and -incompetent molecules and uncoupling of the VSR and particle assembly capacities. The engineered mutants were used to assess the importance of p37 functions for viral infection and the relative contribution of diverse molecular interactions to suppressor activity. Two main conclusions can be drawn: (i) the silencing suppression and encapsidation functions of p37 are both required for systemic Pelargonium line pattern virus infection, and (ii) the suppressor activity of p37 relies on the ability to bind sRNAs rather than on interaction with AGOs. The data also caution against potential misinterpretations of results due to overlap of sequence signals related to distinct protein properties. This is well illustrated by mutation of the GW motif in p37 that concurrently affects nucleolar localization, efficient interaction with AGO1, and sRNA binding capability. These concomitant effects could have been overlooked in other GW motif-containing suppressors, as we exemplify with the orthologous p38 of turnip crinkle virus. PMID:25505185

  4. Key importance of small RNA binding for the activity of a glycine-tryptophan (GW) motif-containing viral suppressor of RNA silencing.

    PubMed

    Pérez-Cañamás, Miryam; Hernández, Carmen

    2015-01-30

    Viruses express viral suppressors of RNA silencing (VSRs) to counteract RNA silencing-based host defenses. Although virtually all stages of the antiviral silencing pathway can be inhibited by VSRs, small RNAs (sRNAs) and Argonaute (AGO) proteins seem to be the most frequent targets. Recently, GW/WG motifs of some VSRs have been proposed to dictate their suppressor function by mediating interaction with AGO(s). Here we have studied the VSR encoded by Pelargonium line pattern virus (family Tombusviridae). The results show that p37, the viral coat protein, blocks RNA silencing. Site-directed mutagenesis of some p37 sequence traits, including a conserved GW motif, allowed generation of suppressor-competent and -incompetent molecules and uncoupling of the VSR and particle assembly capacities. The engineered mutants were used to assess the importance of p37 functions for viral infection and the relative contribution of diverse molecular interactions to suppressor activity. Two main conclusions can be drawn: (i) the silencing suppression and encapsidation functions of p37 are both required for systemic Pelargonium line pattern virus infection, and (ii) the suppressor activity of p37 relies on the ability to bind sRNAs rather than on interaction with AGOs. The data also caution against potential misinterpretations of results due to overlap of sequence signals related to distinct protein properties. This is well illustrated by mutation of the GW motif in p37 that concurrently affects nucleolar localization, efficient interaction with AGO1, and sRNA binding capability. These concomitant effects could have been overlooked in other GW motif-containing suppressors, as we exemplify with the orthologous p38 of turnip crinkle virus.

  5. Key Importance of Small RNA Binding for the Activity of a Glycine-Tryptophan (GW) Motif-containing Viral Suppressor of RNA Silencing*

    PubMed Central

    Pérez-Cañamás, Miryam; Hernández, Carmen

    2015-01-01

    Viruses express viral suppressors of RNA silencing (VSRs) to counteract RNA silencing-based host defenses. Although virtually all stages of the antiviral silencing pathway can be inhibited by VSRs, small RNAs (sRNAs) and Argonaute (AGO) proteins seem to be the most frequent targets. Recently, GW/WG motifs of some VSRs have been proposed to dictate their suppressor function by mediating interaction with AGO(s). Here we have studied the VSR encoded by Pelargonium line pattern virus (family Tombusviridae). The results show that p37, the viral coat protein, blocks RNA silencing. Site-directed mutagenesis of some p37 sequence traits, including a conserved GW motif, allowed generation of suppressor-competent and -incompetent molecules and uncoupling of the VSR and particle assembly capacities. The engineered mutants were used to assess the importance of p37 functions for viral infection and the relative contribution of diverse molecular interactions to suppressor activity. Two main conclusions can be drawn: (i) the silencing suppression and encapsidation functions of p37 are both required for systemic Pelargonium line pattern virus infection, and (ii) the suppressor activity of p37 relies on the ability to bind sRNAs rather than on interaction with AGOs. The data also caution against potential misinterpretations of results due to overlap of sequence signals related to distinct protein properties. This is well illustrated by mutation of the GW motif in p37 that concurrently affects nucleolar localization, efficient interaction with AGO1, and sRNA binding capability. These concomitant effects could have been overlooked in other GW motif-containing suppressors, as we exemplify with the orthologous p38 of turnip crinkle virus. PMID:25505185

  6. Boosting BCG-primed responses with a subunit Apa vaccine during the waning phase improves immunity and imparts protection against Mycobacterium tuberculosis.

    PubMed

    Nandakumar, Subhadra; Kannanganat, Sunil; Dobos, Karen M; Lucas, Megan; Spencer, John S; Amara, Rama Rao; Plikaytis, Bonnie B; Posey, James E; Sable, Suraj B

    2016-01-01

    Heterologous prime-boosting has emerged as a powerful vaccination approach against tuberculosis. However, optimal timing to boost BCG-immunity using subunit vaccines remains unclear in clinical trials. Here, we followed the adhesin Apa-specific T-cell responses in BCG-primed mice and investigated its BCG-booster potential. The Apa-specific T-cell response peaked 32-52 weeks after parenteral or mucosal BCG-priming but waned significantly by 78 weeks. A subunit-Apa-boost during the contraction-phase of BCG-response had a greater effect on the magnitude and functional quality of specific cellular and humoral responses compared to a boost at the peak of BCG-response. The cellular response increased following mucosal BCG-prime-Apa-subunit-boost strategy compared to Apa-subunit-prime-BCG-boost approach. However, parenteral BCG-prime-Apa-subunit-boost by a homologous route was the most effective strategy in-terms of enhancing specific T-cell responses during waning in the lung and spleen. Two Apa-boosters markedly improved waning BCG-immunity and significantly reduced Mycobacterium tuberculosis burdens post-challenge. Our results highlight the challenges of optimization of prime-boost regimens in mice where BCG drives persistent immune-activation and suggest that boosting with a heterologous vaccine may be ideal once the specific persisting effector responses are contracted. Our results have important implications for design of prime-boost regimens against tuberculosis in humans. PMID:27173443

  7. Boosting BCG-primed responses with a subunit Apa vaccine during the waning phase improves immunity and imparts protection against Mycobacterium tuberculosis.

    PubMed

    Nandakumar, Subhadra; Kannanganat, Sunil; Dobos, Karen M; Lucas, Megan; Spencer, John S; Amara, Rama Rao; Plikaytis, Bonnie B; Posey, James E; Sable, Suraj B

    2016-05-13

    Heterologous prime-boosting has emerged as a powerful vaccination approach against tuberculosis. However, optimal timing to boost BCG-immunity using subunit vaccines remains unclear in clinical trials. Here, we followed the adhesin Apa-specific T-cell responses in BCG-primed mice and investigated its BCG-booster potential. The Apa-specific T-cell response peaked 32-52 weeks after parenteral or mucosal BCG-priming but waned significantly by 78 weeks. A subunit-Apa-boost during the contraction-phase of BCG-response had a greater effect on the magnitude and functional quality of specific cellular and humoral responses compared to a boost at the peak of BCG-response. The cellular response increased following mucosal BCG-prime-Apa-subunit-boost strategy compared to Apa-subunit-prime-BCG-boost approach. However, parenteral BCG-prime-Apa-subunit-boost by a homologous route was the most effective strategy in-terms of enhancing specific T-cell responses during waning in the lung and spleen. Two Apa-boosters markedly improved waning BCG-immunity and significantly reduced Mycobacterium tuberculosis burdens post-challenge. Our results highlight the challenges of optimization of prime-boost regimens in mice where BCG drives persistent immune-activation and suggest that boosting with a heterologous vaccine may be ideal once the specific persisting effector responses are contracted. Our results have important implications for design of prime-boost regimens against tuberculosis in humans.

  8. Boosting BCG-primed responses with a subunit Apa vaccine during the waning phase improves immunity and imparts protection against Mycobacterium tuberculosis

    PubMed Central

    Nandakumar, Subhadra; Kannanganat, Sunil; Dobos, Karen M.; Lucas, Megan; Spencer, John S.; Amara, Rama Rao; Plikaytis, Bonnie B.; Posey, James E.; Sable, Suraj B.

    2016-01-01

    Heterologous prime–boosting has emerged as a powerful vaccination approach against tuberculosis. However, optimal timing to boost BCG-immunity using subunit vaccines remains unclear in clinical trials. Here, we followed the adhesin Apa-specific T-cell responses in BCG-primed mice and investigated its BCG-booster potential. The Apa-specific T-cell response peaked 32–52 weeks after parenteral or mucosal BCG-priming but waned significantly by 78 weeks. A subunit-Apa-boost during the contraction-phase of BCG-response had a greater effect on the magnitude and functional quality of specific cellular and humoral responses compared to a boost at the peak of BCG-response. The cellular response increased following mucosal BCG-prime–Apa-subunit-boost strategy compared to Apa-subunit-prime–BCG-boost approach. However, parenteral BCG-prime–Apa-subunit-boost by a homologous route was the most effective strategy in-terms of enhancing specific T-cell responses during waning in the lung and spleen. Two Apa-boosters markedly improved waning BCG-immunity and significantly reduced Mycobacterium tuberculosis burdens post-challenge. Our results highlight the challenges of optimization of prime–boost regimens in mice where BCG drives persistent immune-activation and suggest that boosting with a heterologous vaccine may be ideal once the specific persisting effector responses are contracted. Our results have important implications for design of prime–boost regimens against tuberculosis in humans. PMID:27173443

  9. Interaction of water activity and bicarbonate salts in the inhibition of growth and mycotoxin production by Fusarium and Aspergillus species of importance to corn.

    PubMed

    Samapundo, S; Devlieghere, F; De Meulenaer, B; Lamboni, Y; Osei-Nimoh, D; Debevere, J M

    2007-05-10

    The combined effects of water activity (a(w)) and ammonium/sodium bicarbonate on growth and mycotoxin production in corn by Fusarium and Aspergillus species were investigated. Interaction was observed between the salts and a(w) on the colony growth rates and lag phase durations of all isolates. Growth stimulation at low salt levels was observed only for the Fusarium isolates as the fastest growth of F. verticillioides and F. proliferatum occurred at levels of 0.1-0.2 and 0.5% ammonium and sodium bicarbonate, respectively. Although the complete inhibition of the growth of the Fusarium and Aspergillus isolates investigated took place at a level of 1% ammonium bicarbonate as much as 4% sodium bicarbonate failed to completely inhibit the growth of the Aspergillus isolates. Increase in concentration of either salt generally resulted in large reductions of both fumonisin B(1) and aflatoxin B(1) production. According to the sensorial analysis performed, corn treated with up to 1% ammonium bicarbonate was still acceptable for consumption, whereas corn treated with at least 2% sodium bicarbonate was determined to be sensorially unsuitable. Ammonium bicarbonate can be concluded to be more suitable for protecting stored corn from fungal contamination as it was capable of completely inhibiting both growth and mycotoxin production of the Fusarium and Aspergillus isolates of most importance to corn at levels that were still sensorially acceptable. Therefore ammonium bicarbonate could possibly be applied as a cheap and easy to apply treatment for use in resource limited developing countries.

  10. Aspergillus niger lipase: Heterologous expression in Pichia pastoris, molecular modeling prediction and the importance of the hinge domains at both sides of the lid domain to interfacial activation.

    PubMed

    Shu, Zhengyu; Duan, Mojie; Yang, Jiangke; Xu, Li; Yan, Yunjun

    2009-01-01

    Aspergillus niger lipase (ANL) is an important biocatalyst in the food processing industry. However, there is no report of its detailed three-dimensional structure because of difficulties in crystallization. In this article, based on experimental data and bioinformational analysis results, the structural features of ANL were simulated. Firstly, two recombinant ANLs expressed in Pichia pastoris were purified to homogeneity and their corresponding secondary structure compositions were determined by circular dichroism spectra. Secondly, the primary structure, the secondary structure and the three-dimensional structure of ANL were modeled by comparison with homologous lipases with known three-dimensional structures using the BioEdit software, lipase engineering database (http://www.led.uni-stuttgart.de/), PSIPRED server and SwissModel server. The predicted molecular structure of ANL presented typical features of the alpha/beta hydrolase fold including positioning of the putative catalytic triad residues and the GXSXG signature motif. Comparison of the predicted three-dimensional structure of ANL with the X-ray three-dimensional structure of A. niger feruloyl esterase showed that the functional difference of interfacial activation between lipase and esterase was concerned with the difference in position of the lid. Our three-dimensional model of ANL helps to modify lipase structure by protein engineering, which will further expand the scope of application of ANL. PMID:19248178

  11. Comparative activity of TiO2 microspheres and P25 powder for organic degradation: Implicative importance of structural defects and organic adsorption

    NASA Astrophysics Data System (ADS)

    Wang, Chuan; Liu, Hong; Liu, Yuan; He, Guang'an; Jiang, Chengchun

    2014-11-01

    TiO2 microspheres have been employed as a promisingly new photocatalyst for water and wastewater treatment. P25 TiO2 is commonly employed and its properties are well established as photocatalyst. In this study, photocatalytic activities of the two TiO2 samples are compared by degrading sulfosalicylic acid (SSA), phenol, and 2,4-Dichlorophenoxyacetic acid (2,4-D) under 365 nm UV illumination in a suspension system at neutral pH and associated optimized TiO2 dosages. The results showed that the three organic compounds unexceptionally degraded more rapidly on P25 than on TiO2 microspheres in terms of the concentration-time curves and total organic carbon removals at 120 min. This might me attributed the presence of oxygen vacancies and Ti(III) defects already present on P25 as determined by electron paramagnetic resonance, implying that the defects played an important role for the enhancement of the charge transfer step as rate-determining step. The degradations of three organic compounds on P25 and TiO2 microspheres could be well described by the first-order rate equation, while the degradation kinetics of SSA on TiO2 microspheres was quite different. The difference was ascribed to the medium adsorption ability of SSA on the TiO2 surface.

  12. What Is Meant by "Active Learning?"

    ERIC Educational Resources Information Center

    Petress, Ken

    2008-01-01

    This article compares and contrasts active learning and passive learning. The author describes passive learning as being dependent on a teacher imparting what is to be learned, with little student involvement. Active learning, on the other hand, is a process where students take a dynamic and energetic role in their own education, thereby making…

  13. Effect of the permafrost thawing on the organic carbon and microbial activity in thermokarst lakes of Western Siberia: important source of carbon dioxide in the atmosphere

    NASA Astrophysics Data System (ADS)

    Shirokova, L. S.; Pokrovsky, O. S.; Kirpotin, S. N.; Dupre, B.

    2008-12-01

    "khasyrei" exhibits the highest pH (5.5-6.0), presence of green algae and plants, very low conductivity (4-9 μS) and the lowest OC concentrations. However, there is still quite weak primary productivity (˜ 0.01 mg/C/L) and significant mineralization rate (0.3 mg C/L/day). The bacterial biomass and the number of colony-forming units for different nutrient media demonstrate clear decrease with the temporal evolution of the lake from dystrophic (7000 CFU/mL) at the begging of permafrost thawing to oligotrophic/mesotrophic at the final "khasyrey" stage (80 CFU/mL). Two main conclusions can be drawn from this work: 1) At all stages of lake evolution, there is always high amount of active OM-mineralizing heterotrophic bacteria, and 2) mineralization processes by these microorganisms clearly dominate any possible CO2 consumption by the phytoplankton. As a result, Western Siberia thermokarstic lakes are likely to act as important source of CO2 to the atmosphere. The CO2 flux associated with this microbial activity (i.e., 400 +/- 200 t/km2/y, assuming average lake depth of 1 m) is at least one order of magnitude higher than the riverine DOC flux, bearing in mind very high proportion of newly formed lakes in the Western Siberia (up to 30 percent).

  14. Importance of the short cytoplasmic domain of the feline immunodeficiency virus transmembrane glycoprotein for fusion activity and envelope glycoprotein incorporation into virions

    SciTech Connect

    Celma, Cristina C.P.; Paladino, Monica G.; Gonzalez, Silvia A.; Affranchino, Jose L.

    2007-09-30

    The mature form of the envelope (Env) glycoprotein of lentiviruses is a heterodimer composed of the surface (SU) and transmembrane (TM) subunits. Feline immunodeficiency virus (FIV) possesses a TM glycoprotein with a cytoplasmic tail of approximately 53 amino acids which is unusually short compared with that of the other lentiviral glycoproteins (more than 100 residues). To investigate the relevance of the FIV TM cytoplasmic domain to Env-mediated viral functions, we characterized the biological properties of a series of Env glycoproteins progressively shortened from the carboxyl terminus. All the mutant Env proteins were efficiently expressed in feline cells and processed into the SU and TM subunits. Deletion of 5 or 11 amino acids from the TM C-terminus did not significantly affect Env surface expression, fusogenic activity or Env incorporation into virions, whereas removal of 17 or 23 residues impaired Env-mediated cell-to-cell fusion. Further truncation of the FIV TM by 29 residues resulted in an Env glycoprotein that was poorly expressed at the cell surface, exhibited only 20% of the wild-type Env fusogenic capacity and was inefficiently incorporated into virions. Remarkably, deletion of the TM C-terminal 35 or 41 amino acids restored or even enhanced Env biological functions. Indeed, these mutant Env glycoproteins bearing cytoplasmic domains of 18 or 12 amino acids were found to be significantly more fusogenic than the wild-type Env and were efficiently incorporated into virions. Interestingly, truncation of the TM cytoplasmic domain to only 6 amino acids did not affect Env incorporation into virions but abrogated Env fusogenicity. Finally, removal of the entire TM cytoplasmic tail or deletion of as many as 6 amino acids into the membrane-spanning domain led to a complete loss of Env functions. Our results demonstrate that despite its relatively short length, the FIV TM cytoplasmic domain plays an important role in modulating Env-mediated viral functions.

  15. Antimicrobial peptides: a review of how peptide structure impacts antimicrobial activity

    NASA Astrophysics Data System (ADS)

    Soares, Jason W.; Mello, Charlene M.

    2004-03-01

    Antimicrobial peptides (AMPs) have been discovered in insects, mammals, reptiles, and plants to protect against microbial infection. Many of these peptides have been isolated and studied exhaustively to decipher the molecular mechanisms that impart protection against infectious bacteria, fungi, and viruses. Unfortunately, the molecular mechanisms are still being debated within the scientific community but valuable clues have been obtained through structure/function relationship studies1. Biophysical studies have revealed that cecropins, isolated from insects and pigs, exhibit random structure in solution but undergo a conformational change to an amphipathic α-helix upon interaction with a membrane surface2. The lack of secondary structure in solution results in an extremely durable peptide able to survive exposure to high temperatures, organic solvents and incorporation into fibers and films without compromising antibacterial activity. Studies to better understand the antimicrobial action of cecropins and other AMPs have provided insight into the importance of peptide sequence and structure in antimicrobial activities. Therefore, enhancing our knowledge of how peptide structure imparts function may result in customized peptide sequences tailored for specific applications such as targeted cell delivery systems, novel antibiotics and food preservation additives. This review will summarize the current state of knowledge with respect to cell binding and antimicrobial activity of AMPs focusing primarily upon cecropins.

  16. Activation Mechanism of Oncogenic Deletion Mutations in BRAF, EGFR, and HER2.

    PubMed

    Foster, Scott A; Whalen, Daniel M; Özen, Ayşegül; Wongchenko, Matthew J; Yin, JianPing; Yen, Ivana; Schaefer, Gabriele; Mayfield, John D; Chmielecki, Juliann; Stephens, Philip J; Albacker, Lee A; Yan, Yibing; Song, Kyung; Hatzivassiliou, Georgia; Eigenbrot, Charles; Yu, Christine; Shaw, Andrey S; Manning, Gerard; Skelton, Nicholas J; Hymowitz, Sarah G; Malek, Shiva

    2016-04-11

    Activating mutations in protein kinases drive many cancers. While how recurring point mutations affect kinase activity has been described, the effect of in-frame deletions is not well understood. We show that oncogenic deletions within the β3-αC loop of HER2 and BRAF are analogous to the recurrent EGFR exon 19 deletions. We identify pancreatic carcinomas with BRAF deletions mutually exclusive with KRAS mutations. Crystal structures of BRAF deletions reveal the truncated loop restrains αC in an active "in" conformation, imparting resistance to inhibitors like vemurafenib that bind the αC "out" conformation. Characterization of loop length explains the prevalence of five amino acid deletions in BRAF, EGFR, and HER2 and highlights the importance of this region for kinase activity and inhibitor efficacy. PMID:26996308

  17. A QSAR study of radical scavenging antioxidant activity of a series of flavonoids using DFT based quantum chemical descriptors--the importance of group frontier electron density.

    PubMed

    Sarkar, Ananda; Middya, Tapas Ranjan; Jana, Atish Dipnakar

    2012-06-01

    In a pursuit of electronic level understanding of the antioxidant activity of a series of flavonoids, quantitative structure-activity relationship (QSAR) studies have been carried out using density functional theory (DFT) based quantum chemical descriptors. The best QSAR model have been selected for which the computed square correlation coefficient r(2) = 0.937 and cross-validated squared correlation coefficient q(2) =0.916. The QSAR model indicates that hardness (η), group electrophilic frontier electron density (F(E)(A)) and group philicity (ω(B)(+)) of individual molecules are responsible for in vitro biological activity. To the best our knowledge, the group electrophilic frontier electron density (F(E)(A)) has been used for the first time to explain the radical scavenging activity (RSA) of flavonoids. The excellent correlation between the RSA and the above mentioned DFT based descriptors lead us to predict new antioxidants having very good antioxidant activity.

  18. "We will remember them": a mixed-method study to explore which post-funeral remembrance activities are most significant and important to bereaved people living with loss, and why those particular activities are chosen.

    PubMed

    Vale-Taylor, P

    2009-09-01

    In an increasingly secular age in which society no longer offers a code of behaviour for those who are bereaved as in Victorian times, the majority of people do not seek support from church-based rituals of remembrance. Most hospices provide religious Services of Remembrance and Thanksgiving or non-faith remembrance gatherings for families and friends, and although these are usually well attended, the average number of families represented is usually less than 20% raising the question of whether alternative support should be offered to the remaining majority of families. This study explored which post-funeral remembrance activities are most significant and important to people living with death-related loss, and why those particular activities are chosen. A total of 43 participants took part in a mixed-method study using 2 different data sets: a self-report questionnaire and semi-structured interviews. To add further contextual data to support the study, a third dataset was created when six hospice bereavement counsellors met as a Focus Group. Results indicated that although formal remembrance events are valued, informal rituals created by the bereaved are more important and significant to them. Results could be divided into four different categories: rituals to maintain a 'direct link', or those undertaken 'for' the deceased, rituals that remember the deceased within the community and those viewed as an act of remembrance. The most common reason for choosing a ritual was to keep a bond with the deceased or ensure that the deceased was remembered by others. Remembrance and ritual is personal to each individual and is dynamic, altering from day to day. Remembrance appears to be a journey made up of many small daily rituals, some of which are generic to bereaved people and some of which are highly individualistic. For the majority of people, their informal rituals are far more important than the large planned events that hospices offer because the informal rituals

  19. "We will remember them": a mixed-method study to explore which post-funeral remembrance activities are most significant and important to bereaved people living with loss, and why those particular activities are chosen.

    PubMed

    Vale-Taylor, P

    2009-09-01

    In an increasingly secular age in which society no longer offers a code of behaviour for those who are bereaved as in Victorian times, the majority of people do not seek support from church-based rituals of remembrance. Most hospices provide religious Services of Remembrance and Thanksgiving or non-faith remembrance gatherings for families and friends, and although these are usually well attended, the average number of families represented is usually less than 20% raising the question of whether alternative support should be offered to the remaining majority of families. This study explored which post-funeral remembrance activities are most significant and important to people living with death-related loss, and why those particular activities are chosen. A total of 43 participants took part in a mixed-method study using 2 different data sets: a self-report questionnaire and semi-structured interviews. To add further contextual data to support the study, a third dataset was created when six hospice bereavement counsellors met as a Focus Group. Results indicated that although formal remembrance events are valued, informal rituals created by the bereaved are more important and significant to them. Results could be divided into four different categories: rituals to maintain a 'direct link', or those undertaken 'for' the deceased, rituals that remember the deceased within the community and those viewed as an act of remembrance. The most common reason for choosing a ritual was to keep a bond with the deceased or ensure that the deceased was remembered by others. Remembrance and ritual is personal to each individual and is dynamic, altering from day to day. Remembrance appears to be a journey made up of many small daily rituals, some of which are generic to bereaved people and some of which are highly individualistic. For the majority of people, their informal rituals are far more important than the large planned events that hospices offer because the informal rituals

  20. Controlling the corrosion and cathodic activation of magnesium via microalloying additions of Ge

    NASA Astrophysics Data System (ADS)

    Liu, R. L.; Hurley, M. F.; Kvryan, A.; Williams, G.; Scully, J. R.; Birbilis, N.

    2016-06-01

    The evolution of corrosion morphology and kinetics for magnesium (Mg) have been demonstrated to be influenced by cathodic activation, which implies that the rate of the cathodic partial reaction is enhanced as a result of anodic dissolution. This phenomenon was recently demonstrated to be moderated by the use of arsenic (As) alloying as a poison for the cathodic reaction, leading to significantly improved corrosion resistance. The pursuit of alternatives to toxic As is important as a means to imparting a technologically safe and effective corrosion control method for Mg (and its alloys). In this work, Mg was microalloyed with germanium (Ge), with the aim of improving corrosion resistance by retarding cathodic activation. Based on a combined analysis herein, we report that Ge is potent in supressing the cathodic hydrogen evolution reaction (reduction of water) upon Mg, improving corrosion resistance. With the addition of Ge, cathodic activation of Mg subject to cyclic polarisation was also hindered, with beneficial implications for future Mg electrodes.

  1. Quantification of pulmonary thallium-201 activity after upright exercise in normal persons: importance of peak heart rate and propranolol usage in defining normal values

    SciTech Connect

    Brown, K.A.; Boucher, C.A.; Okada, R.D.; Strauss, H.W.; Pohost, G.M.

    1984-06-01

    Fifty-nine normal patients (34 angiographically normal and 25 clinically normal by Bayesian analysis) underwent thallium-201 imaging after maximal upright exercise. Lung activity was quantitated relative to myocardial activity and a lung/myocardial activity ratio was determined for each patient. Stepwise regression analysis was then used to examine the influence of patient clinical characteristics and exercise variables on the lung/myocardium ratio. Peak heart rate during exercise and propranolol usage both showed significant negative regression coefficients (p less than 0.001). No other patient data showed a significant relation. Using the regression equation and the estimated variance, a 95% confidence level upper limit of normal could be determined for a give peak heart rate and propranolol status. Sixty-one other patients were studied to validate the predicted upper limits of normal based on this model. None of the 27 patients without coronary artery disease had an elevated lung/myocardial ratio, compared with 1 of 8 with 1-vessel disease (difference not significant), 6 of 14 with 2-vessel disease (p less than 0.005), and 6 of 12 with 3-vessel disease (p less than 0.0001). Thus, lung activity on upright exercise thallium-201 studies can be quantitated relative to myocardial activity, and is inversely related to peak heart rate and propranolol use. Use of a regression analysis allows determination of a 95% confidence upper limit of normal to be anticipated in an individual patient.

  2. Hydrogenation of CO2 to methanol: Importance of metal–oxide and metal–carbide interfaces in the activation of CO2

    DOE PAGES

    Rodriguez, José A.; Liu, Ping; Stacchiola, Dario J.; Senanayake, Sanjaya D.; White, Michael G.; Chen, Jingguang G.

    2015-09-30

    In this study, the high thermochemical stability of CO2 makes it very difficult to achieve the catalytic conversion of the molecule into alcohols or other hydrocarbon compounds, which can be used as fuels or the starting point for the generation of fine chemicals. Pure metals and bimetallic systems used for the CO2 → CH3OH conversion usually bind CO2 too weakly and, thus, show low catalytic activity. Here, we discuss a series of recent studies that illustrate the advantages of metal–oxide and metal–carbide interfaces when aiming at the conversion of CO2 into methanol. CeOx/Cu(111), Cu/CeOx/TiO2(110), and Au/CeOx/TiO2(110) exhibit an activity formore » the CO2 → CH3OH conversion that is 2–3 orders of magnitude higher than that of a benchmark Cu(111) catalyst. In the Cu–ceria and Au–ceria interfaces, the multifunctional combination of metal and oxide centers leads to complementary chemical properties that open active reaction pathways for methanol synthesis. Efficient catalysts are also generated after depositing Cu and Au on TiC(001). In these cases, strong metal–support interactions modify the electronic properties of the admetals and make them active for the binding of CO2 and its subsequent transformation into CH3OH at the metal–carbide interfaces.« less

  3. Arabidopsis PHOSPHOTYROSYL PHOSPHATASE ACTIVATOR is essential for PROTEIN PHOSPHATASE 2A holoenzyme assembly and plays important roles in hormone signaling, salt stress response, and plant development.

    PubMed

    Chen, Jian; Hu, Rongbin; Zhu, Yinfeng; Shen, Guoxin; Zhang, Hong

    2014-11-01

    PROTEIN PHOSPHATASE 2A (PP2A) is a major group of serine/threonine protein phosphatases in eukaryotes. It is composed of three subunits: scaffolding subunit A, regulatory subunit B, and catalytic subunit C. Assembly of the PP2A holoenzyme in Arabidopsis (Arabidopsis thaliana) depends on Arabidopsis PHOSPHOTYROSYL PHOSPHATASE ACTIVATOR (AtPTPA). Reduced expression of AtPTPA leads to severe defects in plant development, altered responses to abscisic acid, ethylene, and sodium chloride, and decreased PP2A activity. In particular, AtPTPA deficiency leads to decreased methylation in PP2A-C subunits (PP2Ac). Complete loss of PP2Ac methylation in the suppressor of brassinosteroid insensitive1 mutant leads to 30% reduction of PP2A activity, suggesting that PP2A with a methylated C subunit is more active than PP2A with an unmethylated C subunit. Like AtPTPA, PP2A-A subunits are also required for PP2Ac methylation. The interaction between AtPTPA and PP2Ac is A subunit dependent. In addition, AtPTPA deficiency leads to reduced interactions of B subunits with C subunits, resulting in reduced functional PP2A holoenzyme formation. Thus, AtPTPA is a critical factor for committing the subunit A/subunit C dimer toward PP2A heterotrimer formation.

  4. Validity, Responsiveness, Minimal Detectable Change, and Minimal Clinically Important Change of the Pediatric Motor Activity Log in Children with Cerebral Palsy

    ERIC Educational Resources Information Center

    Lin, Keh-chung; Chen, Hui-fang; Chen, Chia-ling; Wang, Tien-ni; Wu, Ching-yi; Hsieh, Yu-wei; Wu, Li-ling

    2012-01-01

    This study examined criterion-related validity and clinimetric properties of the Pediatric Motor Activity Log (PMAL) in children with cerebral palsy. Study participants were 41 children (age range: 28-113 months) and their parents. Criterion-related validity was evaluated by the associations between the PMAL and criterion measures at baseline and…

  5. Diet and gender are important factors modulating low frequency EEG activity during processing of language sounds in 3 month old infants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Little is known about how early postnatal diet affects brain processes related to cognitive function in healthy infants. To address this question we examined EEG activity recorded from 3 month old infants [breastfed (BF: n = 104, 55 males), milk-based formula fed (MF: n = 114, 57 males) or soy for...

  6. Carbonic anhydrase activity monitored in vivo by hyperpolarized 13C-magnetic resonance spectroscopy demonstrate its importance for pH regulation in tumors

    PubMed Central

    Gallagher, Ferdia A.; Sladen, Helen; Kettunen, Mikko I.; Serrao, Eva M.; Rodrigues, Tiago B.; Wright, Alan; Gill, Andrew B.; McGuire, Sarah; Booth, Thomas C.; Boren, Joan; McIntyre, Alan; Miller, Jodi L.; Lee, Shen-Han; Honess, Davina; Day, Sam E.; Hu, De-en; Howat, William J.; Harris, Adrian L.; Brindle, Kevin M.

    2015-01-01

    Carbonic anhydrase (CA) buffers tissue pH by catalyzing the rapid interconversion of carbon dioxide (CO2) and bicarbonate (HCO3−). We assessed the functional activity of CAIX in two colorectal tumor models, expressing different levels of the enzyme, by measuring the rate of exchange of hyperpolarized 13C label between bicarbonate (H13CO3−) and carbon dioxide (13CO2), following injection of hyperpolarized H13CO3−, using 13C magnetic resonance spectroscopy (13C-MRS) magnetization transfer measurements. 31P-MRS measurements of the chemical shift of the pH probe, 3-aminopropylphosphonate, and 13C-MRS measurements of the H13CO3−/13CO2 peak intensity ratio showed that CAIX overexpression lowered extracellular pH in these tumors. However, the 13C measurements overestimated pH due to incomplete equilibration of the hyperpolarized 13C label between the H13CO3− and 13CO2 pools. Paradoxically, tumors overexpressing CAIX showed lower enzyme activity using magnetization transfer measurements, which can be explained by the more acidic extracellular pH in these tumors and the decreased activity of the enzyme at low pH. This explanation was confirmed by administration of bicarbonate in the drinking water, which elevates tumor extracellular pH and restored enzyme activity to control levels. These results suggest that CAIX expression is increased in hypoxia to compensate for the decrease in its activity produced by a low extracellular pH, and supports the hypothesis that a major function of CAIX is to lower the extracellular pH. PMID:26249175

  7. Carbonic Anhydrase Activity Monitored In Vivo by Hyperpolarized 13C-Magnetic Resonance Spectroscopy Demonstrates Its Importance for pH Regulation in Tumors.

    PubMed

    Gallagher, Ferdia A; Sladen, Helen; Kettunen, Mikko I; Serrao, Eva M; Rodrigues, Tiago B; Wright, Alan; Gill, Andrew B; McGuire, Sarah; Booth, Thomas C; Boren, Joan; McIntyre, Alan; Miller, Jodi L; Lee, Shen-Han; Honess, Davina; Day, Sam E; Hu, De-En; Howat, William J; Harris, Adrian L; Brindle, Kevin M

    2015-10-01

    Carbonic anhydrase buffers tissue pH by catalyzing the rapid interconversion of carbon dioxide (CO2) and bicarbonate (HCO3 (-)). We assessed the functional activity of CAIX in two colorectal tumor models, expressing different levels of the enzyme, by measuring the rate of exchange of hyperpolarized (13)C label between bicarbonate (H(13)CO3(-)) and carbon dioxide ((13)CO2), following injection of hyperpolarized H(13)CO3(-), using (13)C-magnetic resonance spectroscopy ((13)C-MRS) magnetization transfer measurements. (31)P-MRS measurements of the chemical shift of the pH probe, 3-aminopropylphosphonate, and (13)C-MRS measurements of the H(13)CO3(-)/(13)CO2 peak intensity ratio showed that CAIX overexpression lowered extracellular pH in these tumors. However, the (13)C measurements overestimated pH due to incomplete equilibration of the hyperpolarized (13)C label between the H(13)CO3(-) and (13)CO2 pools. Paradoxically, tumors overexpressing CAIX showed lower enzyme activity using magnetization transfer measurements, which can be explained by the more acidic extracellular pH in these tumors and the decreased activity of the enzyme at low pH. This explanation was confirmed by administration of bicarbonate in the drinking water, which elevated tumor extracellular pH and restored enzyme activity to control levels. These results suggest that CAIX expression is increased in hypoxia to compensate for the decrease in its activity produced by a low extracellular pH and supports the hypothesis that a major function of CAIX is to lower the extracellular pH.

  8. How important is importance for prospective memory? A review

    PubMed Central

    Walter, Stefan; Meier, Beat

    2014-01-01

    Forgetting to carry out an intention as planned can have serious consequences in everyday life. People sometimes even forget intentions that they consider as very important. Here, we review the literature on the impact of importance on prospective memory performance. We highlight different methods used to manipulate the importance of a prospective memory task such as providing rewards, importance relative to other ongoing activities, absolute importance, and providing social motives. Moreover, we address the relationship between importance and other factors known to affect prospective memory and ongoing task performance such as type of prospective memory task (time-, event-, or activity-based), cognitive loads, and processing overlaps. Finally, we provide a connection to motivation, we summarize the effects of task importance and we identify important venues for future research. PMID:25018743

  9. Colonization strategies of Pseudomonas fluorescens Pf0-1: activation of soil-specific genes important for diverse and specific environments

    PubMed Central

    2013-01-01

    Background Pseudomonas fluorescens is a common inhabitant of soil and the rhizosphere environment. In addition to potential applications in biocontrol and bioremediation, P. fluorescens is of interest as a model for studying bacterial survival and fitness in soil. A previous study using in vivo expression technology (IVET) identified 22 genes in P. fluorescens Pf0-1 which are up-regulated during growth in Massachusetts loam soil, a subset of which are important for fitness in soil. Despite this and other information on adaptation to soil, downstream applications such as biocontrol or bioremediation in diverse soils remain underdeveloped. We undertook an IVET screen to identify Pf0-1 genes induced during growth in arid Nevada desert soil, to expand our understanding of growth in soil environments, and examine whether Pf0-1 uses general or soil type-specific mechanisms for success in soil environments. Results Twenty six genes were identified. Consistent with previous studies, these genes cluster in metabolism, information storage/processing, regulation, and ‘hypothetical’, but there was no overlap with Pf0-1 genes induced during growth in loam soil. Mutation of both a putative glutamine synthetase gene (Pfl01_2143) and a gene predicted to specify a component of a type VI secretion system (Pfl01_5595) resulted in a decline in arid soil persistence. When examined in sterile loam soil, mutation of Pfl01_5595 had no discernible impact. In contrast, the Pfl01_2143 mutant was not impaired in persistence in sterile soil, but showed a significant reduction in competitive fitness. Conclusions These data support the conclusion that numerous genes are specifically important for survival and fitness in natural environments, and will only be identified using in vivo approaches. Furthermore, we suggest that a subset of soil-induced genes is generally important in different soils, while others may contribute to success in specific types of soil. The importance of glutamine

  10. West Nile Virus Activity in a Winter Roost of American Crows (Corvus brachyrhynchos): Is Bird-To-Bird Transmission Important in Persistence and Amplification?

    PubMed Central

    Hinton, M. G.; Reisen, W. K.; Wheeler, S. S.; Townsend, A. K.

    2015-01-01

    Since its emergence in North America, West Nile virus (WNV) has had a large impact on equines, humans, and wild bird communities, yet gaps remain in our understanding of how the virus persists at temperate latitudes when winter temperatures preclude virus replication and host-seeking activity by mosquito vectors. Bird-to-bird transmission at large communal American Crow roosts could provide one mechanism for WNV persistence. Herein, we describe seasonal patterns of crow and Culex mosquito abundance, WNV infection rates, and the prevalence of WNV-positive fecal samples at a winter crow roost to test the hypothesis that bird-to-bird transmission allows WNV to persist at winter crow roosts. Samples were collected from large winter crow roosts in the Sacramento Valley of California from January 2013 until August 2014, encompassing two overwintering roost periods. West Nile virus RNA was detected in local crow carcasses in both summer [13/18 (72% WNV positive)] and winter [18/44 (41% WNV positive)] 2013–2014. Winter infections were unlikely to have arisen by recent bites from infected mosquitoes because Culex host-seeking activity was very low in winter and all Culex mosquitoes collected during winter months tested negative for WNV. Opportunities existed for fecal-oral transfer at the overwintering roost: most carcasses that tested positive for WNV had detectable viral RNA in both kidney and cloacal swabs, suggesting that infected crows were shedding virus in their feces, and >50% of crows at the roost were stained with feces by mid-winter. Moreover, 2.3% of fecal samples collected in late summer, when mosquitoes were active, tested positive for WNV RNA. Nevertheless, none of the 1,119 feces collected from three roosts over two winters contained detectable WNV RNA. This study provided evidence of WNV infection in overwintering American crows without mosquito vector activity, but did not elucidate a mechanism of WNV transmission during winter. PMID:26335475

  11. West Nile Virus Activity in a Winter Roost of American Crows (Corvus brachyrhynchos): Is Bird-To-Bird Transmission Important in Persistence and Amplification?

    PubMed

    Hinton, M G; Reisen, W K; Wheeler, S S; Townsend, A K

    2015-07-01

    Since its emergence in North America, West Nile virus (WNV) has had a large impact on equines, humans, and wild bird communities, yet gaps remain in our understanding of how the virus persists at temperate latitudes when winter temperatures preclude virus replication and host-seeking activity by mosquito vectors. Bird-to-bird transmission at large communal American Crow roosts could provide one mechanism for WNV persistence. Herein, we describe seasonal patterns of crow and Culex mosquito abundance, WNV infection rates, and the prevalence of WNV-positive fecal samples at a winter crow roost to test the hypothesis that bird-to-bird transmission allows WNV to persist at winter crow roosts. Samples were collected from large winter crow roosts in the Sacramento Valley of California from January 2013 until August 2014, encompassing two overwintering roost periods. West Nile virus RNA was detected in local crow carcasses in both summer [13/18 (72% WNV positive)] and winter [18/44 (41% WNV positive)] 2013-2014. Winter infections were unlikely to have arisen by recent bites from infected mosquitoes because Culex host-seeking activity was very low in winter and all Culex mosquitoes collected during winter months tested negative for WNV. Opportunities existed for fecal-oral transfer at the overwintering roost: most carcasses that tested positive for WNV had detectable viral RNA in both kidney and cloacal swabs, suggesting that infected crows were shedding virus in their feces, and >50% of crows at the roost were stained with feces by mid-winter. Moreover, 2.3% of fecal samples collected in late summer, when mosquitoes were active, tested positive for WNV RNA. Nevertheless, none of the 1,119 feces collected from three roosts over two winters contained detectable WNV RNA. This study provided evidence of WNV infection in overwintering American crows without mosquito vector activity, but did not elucidate a mechanism of WNV transmission during winter.

  12. Diet effects on glucose absorption in the small intestine of neonatal calves: importance of intestinal mucosal growth, lactase activity, and glucose transporters.

    PubMed

    Steinhoff-Wagner, Julia; Zitnan, Rudolf; Schönhusen, Ulrike; Pfannkuche, Helga; Hudakova, Monika; Metges, Cornelia C; Hammon, Harald M

    2014-10-01

    Colostrum (C) feeding in neonatal calves improves glucose status and stimulates intestinal absorptive capacity, leading to greater glucose absorption when compared with milk-based formula feeding. In this study, diet effects on gut growth, lactase activity, and glucose transporters were investigated in several gut segments of the small intestine. Fourteen male German Holstein calves received either C of milkings 1, 3, and 5 (d 1, 2, and 3 in milk) or respective formulas (F) twice daily from d 1 to d 3 after birth. Nutrient content, and especially lactose content, of C and respective F were the same. On d 4, calves were fed C of milking 5 or respective F and calves were slaughtered 2h after feeding. Tissue samples from duodenum and proximal, mid-, and distal jejunum were taken to measure villus size and crypt depth, mucosa and brush border membrane vesicles (BBMV) were taken to determine protein content, and mRNA expression and activity of lactase and mRNA expression of sodium-dependent glucose co-transporter-1 (SGLT1) and facilitative glucose transporter (GLUT2) were determined from mucosal tissue. Additionally, protein expression of SGLT1 in BBMV and GLUT2 in crude mucosal membranes and BBMV were determined, as well as immunochemically localized GLUT2 in the intestinal mucosa. Villus circumference, area, and height were greater, whereas crypt depth was smaller in C than in F. Lactase activity tended to be greater in C than in F. Protein expression of SGLT1 was greater in F than in C. Parameters of villus size, lactase activity, SGLT1 protein expression, as well as apical and basolateral GLUT2 localization in the enterocytes differed among gut segments. In conclusion, C feeding, when compared with F feeding, enhances glucose absorption in neonatal calves primarily by stimulating mucosal growth and increasing absorptive capacity in the small intestine, but not by stimulating abundance of intestinal glucose transporters.

  13. Hydrogenation of CO2 to methanol: Importance of metal–oxide and metal–carbide interfaces in the activation of CO2

    SciTech Connect

    Rodriguez, José A.; Liu, Ping; Stacchiola, Dario J.; Senanayake, Sanjaya D.; White, Michael G.; Chen, Jingguang G.

    2015-09-30

    In this study, the high thermochemical stability of CO2 makes it very difficult to achieve the catalytic conversion of the molecule into alcohols or other hydrocarbon compounds, which can be used as fuels or the starting point for the generation of fine chemicals. Pure metals and bimetallic systems used for the CO2 → CH3OH conversion usually bind CO2 too weakly and, thus, show low catalytic activity. Here, we discuss a series of recent studies that illustrate the advantages of metal–oxide and metal–carbide interfaces when aiming at the conversion of CO2 into methanol. CeOx/Cu(111), Cu/CeOx/TiO2(110), and Au/CeOx/TiO2(110) exhibit an activity for the CO2 → CH3OH conversion that is 2–3 orders of magnitude higher than that of a benchmark Cu(111) catalyst. In the Cu–ceria and Au–ceria interfaces, the multifunctional combination of metal and oxide centers leads to complementary chemical properties that open active reaction pathways for methanol synthesis. Efficient catalysts are also generated after depositing Cu and Au on TiC(001). In these cases, strong metal–support interactions modify the electronic properties of the admetals and make them active for the binding of CO2 and its subsequent transformation into CH3OH at the metal–carbide interfaces.

  14. Cool-weather activity of the forensically important hairy maggot blow fly Chrysomya rufifacies (Macquart) (Diptera: Calliphoridae) on carrion in Upstate South Carolina, United States.

    PubMed

    Cammack, Jonathan A; Nelder, Mark P

    2010-02-25

    The hairy maggot blow fly Chrysomya rufifacies (Macquart) (Diptera: Calliphoridae) has expanded its range in the United States since its introduction into Texas (ca. 1980) and has been collected in 15 states. We investigated the bionomics of immature and adult C. rufifacies collected from carcasses of a raccoon Procyon lotor (Linnaeus) and white-tailed deer Odocoileus virginianus Zimmerman in Upstate South Carolina during November 2007, and used these insects to estimate the minimum period of insect activity. Puparia of C. rufifacies were collected from deer carrion; 28% were parasitized by Nasonia vitripennis (Walker) (Hymenoptera: Pteromalidae). The mean daily ambient temperature during this study was 11.4+/-1.02 degrees C, representing the lowest recorded mean temperature for adult activity of C. rufifacies; adults of C. rufifacies were observed flying among the carcasses at 9.0 degrees C. Although C. rufifacies is considered a warm-weather blow fly, researchers should be aware of its activity at suboptimal conditions, behavior that might aid its expansion into more northern areas. PMID:20042303

  15. Cool-weather activity of the forensically important hairy maggot blow fly Chrysomya rufifacies (Macquart) (Diptera: Calliphoridae) on carrion in Upstate South Carolina, United States.

    PubMed

    Cammack, Jonathan A; Nelder, Mark P

    2010-02-25

    The hairy maggot blow fly Chrysomya rufifacies (Macquart) (Diptera: Calliphoridae) has expanded its range in the United States since its introduction into Texas (ca. 1980) and has been collected in 15 states. We investigated the bionomics of immature and adult C. rufifacies collected from carcasses of a raccoon Procyon lotor (Linnaeus) and white-tailed deer Odocoileus virginianus Zimmerman in Upstate South Carolina during November 2007, and used these insects to estimate the minimum period of insect activity. Puparia of C. rufifacies were collected from deer carrion; 28% were parasitized by Nasonia vitripennis (Walker) (Hymenoptera: Pteromalidae). The mean daily ambient temperature during this study was 11.4+/-1.02 degrees C, representing the lowest recorded mean temperature for adult activity of C. rufifacies; adults of C. rufifacies were observed flying among the carcasses at 9.0 degrees C. Although C. rufifacies is considered a warm-weather blow fly, researchers should be aware of its activity at suboptimal conditions, behavior that might aid its expansion into more northern areas.

  16. Modes of Action of the New Arylguanidine Abafungin beyond Interference with Ergosterol Biosynthesis and in vitro Activity against Medically Important Fungi

    PubMed Central

    Borelli, C.; Schaller, M.; Niewerth, M.; Nocker, K.; Baasner, B.; Berg, D.; Tiemann, R.; Tietjen, K.; Fugmann, B.; Lang-Fugmann, S.; Korting, H.C.

    2008-01-01

    Background In contrast to the increasing numbers of agents for the treatment of invasive fungal infections, discoveries of new antifungal agents with therapeutic value in dermatomycoses are reported only rarely. Methods Abafungin (chemical abstracts service registry No. 129639-79/8) is the first member of a novel class of synthetic antifungal compounds, the arylguanidines. It was first synthesized at Bayer AG, Leverkusen, Germany, and its antifungal action was discovered during the screening of H2-receptor antagonists based on the structure of famotidine. To obtain insight into its mode of action and antifungal activity, various tests were carried out with different fungal pathogensin vitro. Results Abafungin was found to have potent antifungal activity. Furthermore, mode-of-action studies suggested that abafungin exerts its antifungal activity regardless of whether the pathogens are growing or in a resting state. One target of abafungin was found to be the inhibition of transmethylation at the C-24 position of the sterol side chain, catalyzed by the enzyme sterol-C-24-methyltransferase. A second action of abafungin seems to be a direct effect on the fungal cell membrane. Conclusion The observed characteristics of abafungin indicate that abafungin might be a promising antifungal agent defining a new class of antimycotics. PMID:18587237

  17. Extracellular enzyme activity and microbial diversity measured on seafloor exposed basalts from Loihi seamount indicate the importance of basalts to global biogeochemical cycling.

    PubMed

    Jacobson Meyers, Myrna E; Sylvan, Jason B; Edwards, Katrina J

    2014-08-01

    Seafloor basalts are widely distributed and host diverse prokaryotic communities, but no data exist concerning the metabolic rates of the resident microbial communities. We present here potential extracellular enzyme activities of leucine aminopeptidase (LAP) and alkaline phosphatase (AP) measured on basalt samples from different locations on Loihi Seamount, HI, coupled with analysis of prokaryotic biomass and pyrosequencing of the bacterial 16S rRNA gene. The community maximum potential enzyme activity (Vmax) of LAP ranged from 0.47 to 0.90 nmol (g rock)(-1) h(-1); the Vmax for AP was 28 to 60 nmol (g rock)(-1) h(-1). The Km of LAP ranged from 26 to 33 μM, while the Km for AP was 2 to 7 μM. Bacterial communities on Loihi basalts were comprised primarily of Alpha-, Delta-, andGammaproteobacteria, Bacteroidetes, and Planctomycetes. The putative ability to produce LAP is evenly distributed across the most commonly detected bacterial orders, but the ability to produce AP is likely dominated by bacteria in the orders Xanthomonadales, Flavobacteriales, and Planctomycetales. The enzyme activities on Loihi basalts were compared to those of other marine environments that have been studied and were found to be similar in magnitude to those from continental shelf sediments and orders of magnitude higher than any measured in the water column, demonstrating that the potential for exposed basalts to transform organic matter is substantial. We propose that microbial communities on basaltic rock play a significant, quantifiable role in benthic biogeochemical processes. PMID:24907315

  18. Probing the importance of hydrogen bonds in the active site of the subtilisin nattokinase by site-directed mutagenesis and molecular dynamics simulation

    PubMed Central

    Zheng, Zhong-liang; Ye, Mao-qing; Zuo, Zhen-yu; Liu, Zhi-gang; Tai, Keng-chang; Zou, Guo-lin

    2006-01-01

    Hydrogen bonds occurring in the catalytic triad (Asp32, His64 and Ser221) and the oxyanion hole (Asn155) are very important to the catalysis of peptide bond hydrolysis by serine proteases. For the subtilisin NK (nattokinase), a bacterial serine protease, construction and analysis of a three-dimensional structural model suggested that several hydrogen bonds formed by four residues function to stabilize the transition state of the hydrolysis reaction. These four residues are Ser33, Asp60, Ser62 and Thr220. In order to remove the effect of these hydrogen bonds, four mutants (Ser33→Ala33, Asp60→Ala60, Ser62→Ala62, and Thr220→Ala220) were constructed by site-directed mutagenesis. The results of enzyme kinetics indicated that removal of these hydrogen bonds increases the free-energy of the transition state (ΔΔGT). We concluded that these hydrogen bonds are more important for catalysis than for binding the substrate, because removal of these bonds mainly affects the kcat but not the Km values. A substrate, SUB1 (succinyl-Ala-Ala-Pro-Phe-p-nitroanilide), was used during enzyme kinetics experiments. In the present study we have also shown the results of FEP (free-energy perturbation) calculations with regard to the binding and catalysis reactions for these mutant subtilisins. The calculated difference in FEP also suggested that these four residues are more important for catalysis than binding of the substrate, and the simulated values compared well with the experimental values from enzyme kinetics. The results of MD (molecular dynamics) simulations further demonstrated that removal of these hydrogen bonds partially releases Asp32, His64 and Asn155 so that the stability of the transition state decreases. Another substrate, SUB2 (H-D-Val-Leu-Lys-p-nitroanilide), was used for FEP calculations and MD simulations. PMID:16411898

  19. Probing the importance of hydrogen bonds in the active site of the subtilisin nattokinase by site-directed mutagenesis and molecular dynamics simulation.

    PubMed

    Zheng, Zhong-liang; Ye, Mao-qing; Zuo, Zhen-yu; Liu, Zhi-gang; Tai, Keng-chang; Zou, Guo-lin

    2006-05-01

    Hydrogen bonds occurring in the catalytic triad (Asp32, His64 and Ser221) and the oxyanion hole (Asn155) are very important to the catalysis of peptide bond hydrolysis by serine proteases. For the subtilisin NK (nattokinase), a bacterial serine protease, construction and analysis of a three-dimensional structural model suggested that several hydrogen bonds formed by four residues function to stabilize the transition state of the hydrolysis reaction. These four residues are Ser33, Asp60, Ser62 and Thr220. In order to remove the effect of these hydrogen bonds, four mutants (Ser33-->Ala33, Asp60-->Ala60, Ser62-->Ala62, and Thr220-->Ala220) were constructed by site-directed mutagenesis. The results of enzyme kinetics indicated that removal of these hydrogen bonds increases the free-energy of the transition state (DeltaDeltaG(T)). We concluded that these hydrogen bonds are more important for catalysis than for binding the substrate, because removal of these bonds mainly affects the kcat but not the K(m) values. A substrate, SUB1 (succinyl-Ala-Ala-Pro-Phe-p-nitroanilide), was used during enzyme kinetics experiments. In the present study we have also shown the results of FEP (free-energy perturbation) calculations with regard to the binding and catalysis reactions for these mutant subtilisins. The calculated difference in FEP also suggested that these four residues are more important for catalysis than binding of the substrate, and the simulated values compared well with the experimental values from enzyme kinetics. The results of MD (molecular dynamics) simulations further demonstrated that removal of these hydrogen bonds partially releases Asp32, His64 and Asn155 so that the stability of the transition state decreases. Another substrate, SUB2 (H-D-Val-Leu-Lys-p-nitroanilide), was used for FEP calculations and MD simulations.

  20. Expression of GhNAC2 from G. herbaceum, improves root growth and imparts tolerance to drought in transgenic cotton and Arabidopsis.

    PubMed

    Gunapati, Samatha; Naresh, Ram; Ranjan, Sanjay; Nigam, Deepti; Hans, Aradhana; Verma, Praveen C; Gadre, Rekha; Pathre, Uday V; Sane, Aniruddha P; Sane, Vidhu A

    2016-01-01

    NAC proteins are plant-specific transcription factors that play essential roles in regulating development and responses to abiotic and biotic stresses. We show that over-expression of the cotton GhNAC2 under the CaMV35S promoter increases root growth in both Arabidopsis and cotton under unstressed conditions. Transgenic Arabidopsis plants also show improved root growth in presence of mannitol and NaCl while transgenic cotton expressing GhNAC2 show reduced leaf abscission and wilting upon water stress compared to control plants. Transgenic Arabidopsis plants also have larger leaves, higher seed number and size under well watered conditions, reduced transpiration and higher relative leaf water content. Micro-array analysis of transgenic plants over-expressing GhNAC2 reveals activation of the ABA/JA pathways and a suppression of the ethylene pathway at several levels to reduce expression of ERF6/ERF1/WRKY33/ MPK3/MKK9/ACS6 and their targets. This probably suppresses the ethylene-mediated inhibition of organ expansion, leading to larger leaves, better root growth and higher yields under unstressed conditions. Suppression of the ethylene pathway and activation of the ABA/JA pathways also primes the plant for improved stress tolerance by reduction in transpiration, greater stomatal control and suppression of growth retarding factors. PMID:27113714

  1. Expression of GhNAC2 from G. herbaceum, improves root growth and imparts tolerance to drought in transgenic cotton and Arabidopsis.

    PubMed

    Gunapati, Samatha; Naresh, Ram; Ranjan, Sanjay; Nigam, Deepti; Hans, Aradhana; Verma, Praveen C; Gadre, Rekha; Pathre, Uday V; Sane, Aniruddha P; Sane, Vidhu A

    2016-04-26

    NAC proteins are plant-specific transcription factors that play essential roles in regulating development and responses to abiotic and biotic stresses. We show that over-expression of the cotton GhNAC2 under the CaMV35S promoter increases root growth in both Arabidopsis and cotton under unstressed conditions. Transgenic Arabidopsis plants also show improved root growth in presence of mannitol and NaCl while transgenic cotton expressing GhNAC2 show reduced leaf abscission and wilting upon water stress compared to control plants. Transgenic Arabidopsis plants also have larger leaves, higher seed number and size under well watered conditions, reduced transpiration and higher relative leaf water content. Micro-array analysis of transgenic plants over-expressing GhNAC2 reveals activation of the ABA/JA pathways and a suppression of the ethylene pathway at several levels to reduce expression of ERF6/ERF1/WRKY33/ MPK3/MKK9/ACS6 and their targets. This probably suppresses the ethylene-mediated inhibition of organ expansion, leading to larger leaves, better root growth and higher yields under unstressed conditions. Suppression of the ethylene pathway and activation of the ABA/JA pathways also primes the plant for improved stress tolerance by reduction in transpiration, greater stomatal control and suppression of growth retarding factors.

  2. Expression of GhNAC2 from G. herbaceum, improves root growth and imparts tolerance to drought in transgenic cotton and Arabidopsis

    PubMed Central

    Gunapati, Samatha; Naresh, Ram; Ranjan, Sanjay; Nigam, Deepti; Hans, Aradhana; Verma, Praveen C.; Gadre, Rekha; Pathre, Uday V.; Sane, Aniruddha P.; Sane, Vidhu A.

    2016-01-01

    NAC proteins are plant-specific transcription factors that play essential roles in regulating development and responses to abiotic and biotic stresses. We show that over-expression of the cotton GhNAC2 under the CaMV35S promoter increases root growth in both Arabidopsis and cotton under unstressed conditions. Transgenic Arabidopsis plants also show improved root growth in presence of mannitol and NaCl while transgenic cotton expressing GhNAC2 show reduced leaf abscission and wilting upon water stress compared to control plants. Transgenic Arabidopsis plants also have larger leaves, higher seed number and size under well watered conditions, reduced transpiration and higher relative leaf water content. Micro-array analysis of transgenic plants over-expressing GhNAC2 reveals activation of the ABA/JA pathways and a suppression of the ethylene pathway at several levels to reduce expression of ERF6/ERF1/WRKY33/ MPK3/MKK9/ACS6 and their targets. This probably suppresses the ethylene-mediated inhibition of organ expansion, leading to larger leaves, better root growth and higher yields under unstressed conditions. Suppression of the ethylene pathway and activation of the ABA/JA pathways also primes the plant for improved stress tolerance by reduction in transpiration, greater stomatal control and suppression of growth retarding factors. PMID:27113714

  3. Tsetse GmmSRPN10 Has Anti-complement Activity and Is Important for Successful Establishment of Trypanosome Infections in the Fly Midgut

    PubMed Central

    Ooi, Cher-Pheng; Haines, Lee R.; Southern, Daniel M.; Lehane, Michael J.; Acosta-Serrano, Alvaro

    2015-01-01

    The complement cascade in mammalian blood can damage the alimentary tract of haematophagous arthropods. As such, these animals have evolved their own repertoire of complement-inactivating factors, which are inadvertently exploited by blood-borne pathogens to escape complement lysis. Unlike the bloodstream stages, the procyclic (insect) stage of Trypanosoma brucei is highly susceptible to complement killing, which is puzzling considering that a tsetse takes a bloodmeal every 2–4 days. In this study, we identified four tsetse (Glossina morsitans morsitans) serine protease inhibitors (serpins) from a midgut expressed sequence tag (EST) library (GmmSRPN3, GmmSRPN5, GmmSRPN9 and GmmSRPN10) and investigated their role in modulating the establishment of a T. brucei infection in the midgut. Although not having evolved in a common blood-feeding ancestor, all four serpins have an active site sharing remarkable homology with the human complement C1-inhibitor serpin, SerpinG1. RNAi knockdown of individual GmmSRPN9 and GmmSRPN10 genes resulted in a significant decreased rate of infection by procyclic form T. brucei. Furthermore, recombinant GmmSRPN10 was both able to inhibit the activity of human complement-cascade serine proteases, C1s and Factor D, and to protect the in vitro killing of procyclic trypanosomes when incubated with complement-activated human serum. Thus, the secretion of serpins, which may be part of a bloodmeal complement inactivation system in tsetse, is used by procyclic trypanosomes to evade an influx of fresh trypanolytic complement with each bloodmeal. This highlights another facet of the complicated relationship between T. brucei and its tsetse vector, where the parasite takes advantage of tsetse physiology to further its chances of propagation and transmission. PMID:25569180

  4. NMR structure of the noncytotoxic α-sarcin mutant Δ(7-22): The importance of the native conformation of peripheral loops for activity

    PubMed Central

    García-Mayoral, Ma Flor; García-Ortega, Lucia; Lillo, Ma Pilar; Santoro, Jorge; Martínez Del Pozo, Álvaro; Gavilanes, José G.; Rico, Manuel; Bruix, Marta

    2004-01-01

    The deletion mutant Δ(7-22) of α-sarcin, unlike its wild-type protein counterpart, lacks the specific ability to degrade rRNA in intact ribosomes and exhibits an increased unspecific ribonuclease activity and decreased interaction with lipid vesicles. In trying to shed light on these differences, we report here on the three-dimensional structure of the Δ(7-22) α-sarcin mutant using NMR methods. We also evaluated its dynamic properties on the basis of theoretical models and measured its correlation time (6.2 nsec) by time-resolved fluorescence anisotropy. The global fold characteristic of ribotoxins is preserved in the mutant. The most significant differences with respect to the α-sarcin structure are concentrated in (1) loop 2, (2) loop 3, which adopts a new orientation, and (3) loop 5, which shows multiple conformations and an altered dynamics. The interactions between loop 5 and the N-terminal hairpin are lost in the mutant, producing increased solvent accessibility of the active-site residues. The degree of solvent exposure of the catalytic His 137 is similar to that shown by His 92 in RNase T1. Additionally, the calculated order parameters of residues belonging to loop 5 in the mutant correspond to an internal dynamic behavior more similar to RNase T1 than α-sarcin. On the other hand, changes in the relative orientation of loop 3 move the lysine-rich region 111–114, crucial for substrate recognition, away from the active site. All of the structural and dynamic data presented here reveal that the mutant is a hybrid of ribotoxins and noncytotoxic ribonucleases, consistent with its biological properties. PMID:15044731

  5. Tsetse GmmSRPN10 has anti-complement activity and is important for successful establishment of trypanosome infections in the fly midgut.

    PubMed

    Ooi, Cher-Pheng; Haines, Lee R; Southern, Daniel M; Lehane, Michael J; Acosta-Serrano, Alvaro

    2015-01-01

    The complement cascade in mammalian blood can damage the alimentary tract of haematophagous arthropods. As such, these animals have evolved their own repertoire of complement-inactivating factors, which are inadvertently exploited by blood-borne pathogens to escape complement lysis. Unlike the bloodstream stages, the procyclic (insect) stage of Trypanosoma brucei is highly susceptible to complement killing, which is puzzling considering that a tsetse takes a bloodmeal every 2-4 days. In this study, we identified four tsetse (Glossina morsitans morsitans) serine protease inhibitors (serpins) from a midgut expressed sequence tag (EST) library (GmmSRPN3, GmmSRPN5, GmmSRPN9 and GmmSRPN10) and investigated their role in modulating the establishment of a T. brucei infection in the midgut. Although not having evolved in a common blood-feeding ancestor, all four serpins have an active site sharing remarkable homology with the human complement C1-inhibitor serpin, SerpinG1. RNAi knockdown of individual GmmSRPN9 and GmmSRPN10 genes resulted in a significant decreased rate of infection by procyclic form T. brucei. Furthermore, recombinant GmmSRPN10 was both able to inhibit the activity of human complement-cascade serine proteases, C1s and Factor D, and to protect the in vitro killing of procyclic trypanosomes when incubated with complement-activated human serum. Thus, the secretion of serpins, which may be part of a bloodmeal complement inactivation system in tsetse, is used by procyclic trypanosomes to evade an influx of fresh trypanolytic complement with each bloodmeal. This highlights another facet of the complicated relationship between T. brucei and its tsetse vector, where the parasite takes advantage of tsetse physiology to further its chances of propagation and transmission. PMID:25569180

  6. Mechanisms Underlying the Delayed Activation of the Cap1 Transcription Factor in Candida albicans following Combinatorial Oxidative and Cationic Stress Important for Phagocytic Potency

    PubMed Central

    Kos, Iaroslava; Patterson, Miranda J.; Znaidi, Sadri; Kaloriti, Despoina; da Silva Dantas, Alessandra; Herrero-de-Dios, Carmen M.; d’Enfert, Christophe; Brown, Alistair J. P.

    2016-01-01

    ABSTRACT Following phagocytosis, microbes are exposed to an array of antimicrobial weapons that include reactive oxygen species (ROS) and cationic fluxes. This is significant as combinations of oxidative and cationic stresses are much more potent than the corresponding single stresses, triggering the synergistic killing of the fungal pathogen Candida albicans by “stress pathway interference.” Previously we demonstrated that combinatorial oxidative plus cationic stress triggers a dramatic increase in intracellular ROS levels compared to oxidative stress alone. Here we show that activation of Cap1, the major regulator of antioxidant gene expression in C. albicans, is significantly delayed in response to combinatorial stress treatments and to high levels of H2O2. Cap1 is normally oxidized in response to H2O2; this masks the nuclear export sequence, resulting in the rapid nuclear accumulation of Cap1 and the induction of Cap1-dependent genes. Here we demonstrate that following exposure of cells to combinatorial stress or to high levels of H2O2, Cap1 becomes trapped in a partially oxidized form, Cap1OX-1. Notably, Cap1-dependent gene expression is not induced when Cap1 is in this partially oxidized form. However, while Cap1OX-1 readily accumulates in the nucleus and binds to target genes following high-H2O2 stress, the nuclear accumulation of Cap1OX-1 following combinatorial H2O2 and NaCl stress is delayed due to a cationic stress-enhanced interaction with the Crm1 nuclear export factor. These findings define novel mechanisms that delay activation of the Cap1 transcription factor, thus preventing the rapid activation of the stress responses vital for the survival of C. albicans within the host. PMID:27025253

  7. Extracellular Enzyme Activity and Microbial Diversity Measured on Seafloor Exposed Basalts from Loihi Seamount Indicate the Importance of Basalts to Global Biogeochemical Cycling

    PubMed Central

    Sylvan, Jason B.; Edwards, Katrina J.

    2014-01-01

    Seafloor basalts are widely distributed and host diverse prokaryotic communities, but no data exist concerning the metabolic rates of the resident microbial communities. We present here potential extracellular enzyme activities of leucine aminopeptidase (LAP) and alkaline phosphatase (AP) measured on basalt samples from different locations on Loihi Seamount, HI, coupled with analysis of prokaryotic biomass and pyrosequencing of the bacterial 16S rRNA gene. The community maximum potential enzyme activity (Vmax) of LAP ranged from 0.47 to 0.90 nmol (g rock)−1 h−1; the Vmax for AP was 28 to 60 nmol (g rock)−1 h−1. The Km of LAP ranged from 26 to 33 μM, while the Km for AP was 2 to 7 μM. Bacterial communities on Loihi basalts were comprised primarily of Alpha-, Delta-, andGammaproteobacteria, Bacteroidetes, and Planctomycetes. The putative ability to produce LAP is evenly distributed across the most commonly detected bacterial orders, but the ability to produce AP is likely dominated by bacteria in the orders Xanthomonadales, Flavobacteriales, and Planctomycetales. The enzyme activities on Loihi basalts were compared to those of other marine environments that have been studied and were found to be similar in magnitude to those from continental shelf sediments and orders of magnitude higher than any measured in the water column, demonstrating that the potential for exposed basalts to transform organic matter is substantial. We propose that microbial communities on basaltic rock play a significant, quantifiable role in benthic biogeochemical processes. PMID:24907315

  8. Identification of Influenza A/PR/8/34 Donor Viruses Imparting High Hemagglutinin Yields to Candidate Vaccine Viruses in Eggs

    PubMed Central

    Winne, Emily; Santana, Wanda; Metcalfe, Maureen G.; Mateu-Petit, Guaniri; Ridenour, Callie; Hossain, M. Jaber; Villanueva, Julie; Zaki, Sherif R.; Williams, Tracie L.; Cox, Nancy J.; Barr, John R.; Donis, Ruben O.

    2015-01-01

    One of the important lessons learned from the 2009 H1N1 pandemic is that a high yield influenza vaccine virus is essential for efficient and timely production of pandemic vaccines in eggs. The current seasonal and pre-pandemic vaccine viruses are generated either by classical reassortment or reverse genetics. Both approaches utilize a high growth virus, generally A/Puerto Rico/8/1934 (PR8), as the donor of all or most of the internal genes, and the wild type virus recommended for inclusion in the vaccine to contribute the hemagglutinin (HA) and neuraminidase (NA) genes encoding the surface glycoproteins. As a result of extensive adaptation through sequential egg passaging, PR8 viruses with different gene sequences and high growth properties have been selected at different laboratories in past decades. The effect of these related but distinct internal PR8 genes on the growth of vaccine viruses in eggs has not been examined previously. Here, we use reverse genetics to analyze systematically the growth and HA antigen yield of reassortant viruses with 3 different PR8 backbones. A panel of 9 different HA/NA gene pairs in combination with each of the 3 different lineages of PR8 internal genes (27 reassortant viruses) was generated to evaluate their performance. Virus and HA yield assays showed that the PR8 internal genes influence HA yields in most subtypes. Although no single PR8 internal gene set outperformed the others in all candidate vaccine viruses, a combination of specific PR8 backbone with individual HA/NA pairs demonstrated improved HA yield and consequently the speed of vaccine production. These findings may be important both for production of seasonal vaccines and for a rapid global vaccine response during a pandemic. PMID:26068666

  9. Mitogen-Activated Protein Kinase Cascade MKK7-MPK6 Plays Important Roles in Plant Development and Regulates Shoot Branching by Phosphorylating PIN1 in Arabidopsis.

    PubMed

    Jia, Weiyan; Li, Baohua; Li, Shujia; Liang, Yan; Wu, Xiaowei; Ma, Mei; Wang, Jiyao; Gao, Jin; Cai, Yueyue; Zhang, Yuanya; Wang, Yingchun; Li, Jiayang; Wang, Yonghong

    2016-09-01

    Emerging evidences exhibit that mitogen-activated protein kinase (MAPK/MPK) signaling pathways are connected with many aspects of plant development. The complexity of MAPK cascades raises challenges not only to identify the MAPK module in planta but also to define the specific role of an individual module. So far, our knowledge of MAPK signaling has been largely restricted to a small subset of MAPK cascades. Our previous study has characterized an Arabidopsis bushy and dwarf1 (bud1) mutant, in which the MAP Kinase Kinase 7 (MKK7) was constitutively activated, resulting in multiple phenotypic alterations. In this study, we found that MPK3 and MPK6 are the substrates for phosphorylation by MKK7 in planta. Genetic analysis showed that MKK7-MPK6 cascade is specifically responsible for the regulation of shoot branching, hypocotyl gravitropism, filament elongation, and lateral root formation, while MKK7-MPK3 cascade is mainly involved in leaf morphology. We further demonstrated that the MKK7-MPK6 cascade controls shoot branching by phosphorylating Ser 337 on PIN1, which affects the basal localization of PIN1 in xylem parenchyma cells and polar auxin transport in the primary stem. Our results not only specify the functions of the MKK7-MPK6 cascade but also reveal a novel mechanism for PIN1 phosphorylation, establishing a molecular link between the MAPK cascade and auxin-regulated plant development. PMID:27618482

  10. Mitogen-Activated Protein Kinase Cascade MKK7-MPK6 Plays Important Roles in Plant Development and Regulates Shoot Branching by Phosphorylating PIN1 in Arabidopsis

    PubMed Central

    Liang, Yan; Wu, Xiaowei; Cai, Yueyue; Zhang, Yuanya; Wang, Yingchun; Li, Jiayang; Wang, Yonghong

    2016-01-01

    Emerging evidences exhibit that mitogen-activated protein kinase (MAPK/MPK) signaling pathways are connected with many aspects of plant development. The complexity of MAPK cascades raises challenges not only to identify the MAPK module in planta but also to define the specific role of an individual module. So far, our knowledge of MAPK signaling has been largely restricted to a small subset of MAPK cascades. Our previous study has characterized an Arabidopsis bushy and dwarf1 (bud1) mutant, in which the MAP Kinase Kinase 7 (MKK7) was constitutively activated, resulting in multiple phenotypic alterations. In this study, we found that MPK3 and MPK6 are the substrates for phosphorylation by MKK7 in planta. Genetic analysis showed that MKK7-MPK6 cascade is specifically responsible for the regulation of shoot branching, hypocotyl gravitropism, filament elongation, and lateral root formation, while MKK7-MPK3 cascade is mainly involved in leaf morphology. We further demonstrated that the MKK7-MPK6 cascade controls shoot branching by phosphorylating Ser 337 on PIN1, which affects the basal localization of PIN1 in xylem parenchyma cells and polar auxin transport in the primary stem. Our results not only specify the functions of the MKK7-MPK6 cascade but also reveal a novel mechanism for PIN1 phosphorylation, establishing a molecular link between the MAPK cascade and auxin-regulated plant development. PMID:27618482

  11. The possibly important role played by Ga{sub 2}O{sub 3} during the activation of GaN photocathode

    SciTech Connect

    Fu, Xiaoqian E-mail: 214808748@qq.com; Wang, Honggang; Zhang, Junju; Li, Zhiming; Cui, Shiyao; Zhang, Lejuan

    2015-08-14

    Three different chemical solutions are used to remove the possible contamination on GaN surface, while Ga{sub 2}O{sub 3} is still found at the surface. After thermal annealing at 710 °C in the ultrahigh vacuum (UHV) chamber and activated with Cs/O, all the GaN samples are successfully activated to the effective negative electron affinity (NEA) photocathodes. Among all samples, the GaN sample with the highest content of Ga{sub 2}O{sub 3} after chemical cleaning obtains the highest quantum efficiency. By analyzing the property of Ga{sub 2}O{sub 3}, the surface processing results, and electron affinity variations during Cs and Cs/O{sub 2} deposition on GaN of other groups, it is suggested that before the adsorption of Cs, Ga{sub 2}O{sub 3} is not completely removed from GaN surface in our samples, which will combine with Cs and lead to a large decrease in electron affinity. Furthermore, the effective NEA is formed for GaN photocathode, along with the surface downward band bending. Based on this assumption, a new dipole model Ga{sub 2}O{sub 3}-Cs is suggested, and the experimental effects are explained and discussed.

  12. Mitogen-Activated Protein Kinase Cascade MKK7-MPK6 Plays Important Roles in Plant Development and Regulates Shoot Branching by Phosphorylating PIN1 in Arabidopsis.

    PubMed

    Jia, Weiyan; Li, Baohua; Li, Shujia; Liang, Yan; Wu, Xiaowei; Ma, Mei; Wang, Jiyao; Gao, Jin; Cai, Yueyue; Zhang, Yuanya; Wang, Yingchun; Li, Jiayang; Wang, Yonghong

    2016-09-01

    Emerging evidences exhibit that mitogen-activated protein kinase (MAPK/MPK) signaling pathways are connected with many aspects of plant development. The complexity of MAPK cascades raises challenges not only to identify the MAPK module in planta but also to define the specific role of an individual module. So far, our knowledge of MAPK signaling has been largely restricted to a small subset of MAPK cascades. Our previous study has characterized an Arabidopsis bushy and dwarf1 (bud1) mutant, in which the MAP Kinase Kinase 7 (MKK7) was constitutively activated, resulting in multiple phenotypic alterations. In this study, we found that MPK3 and MPK6 are the substrates for phosphorylation by MKK7 in planta. Genetic analysis showed that MKK7-MPK6 cascade is specifically responsible for the regulation of shoot branching, hypocotyl gravitropism, filament elongation, and lateral root formation, while MKK7-MPK3 cascade is mainly involved in leaf morphology. We further demonstrated that the MKK7-MPK6 cascade controls shoot branching by phosphorylating Ser 337 on PIN1, which affects the basal localization of PIN1 in xylem parenchyma cells and polar auxin transport in the primary stem. Our results not only specify the functions of the MKK7-MPK6 cascade but also reveal a novel mechanism for PIN1 phosphorylation, establishing a molecular link between the MAPK cascade and auxin-regulated plant development.

  13. The Importance of Toll-like Receptors in NF-κB Signaling Pathway Activation by Helicobacter pylori Infection and the Regulators of this Response.

    PubMed

    Hu, Yi; Liu, Jian-Ping; Zhu, Yin; Lu, Nong-Hua

    2016-10-01

    Helicobacter pylori (H. pylori) is a common pathogenic bacterium in the stomach that infects almost half of the population worldwide and is closely related to gastric diseases and some extragastric diseases, including iron-deficiency anemia and idiopathic thrombocytopenic purpura. Both the Maastricht IV/Florence consensus report and the Kyoto global consensus report have proposed the eradication of H. pylori to prevent gastric cancer as H.pylori has been shown to be a major cause of gastric carcinogenesis. The interactions between H. pylori and host receptors induce the release of the proinflammatory cytokines by activating proinflammatory signaling pathways such as nuclear factor kappa B (NF-κB), which plays a central role in inflammation, immune response, and carcinogenesis. Among these receptors, Toll-like receptors (TLRs) are classical pattern recognition receptors in the recognition of H. pylori and the mediation of the host inflammatory and immune responses to H. pylori. TLR polymorphisms also contribute to the clinical consequences of H. pylori infection. In this review, we focus on the functions of TLRs in the NF-κB signaling pathway activated by H. pylori, the regulators modulating this response, and the functions of TLR polymorphisms in H.pylori-related diseases.

  14. Importance of the operating pH in maintaining the stability of anoxic ammonium oxidation (anammox) activity in moving bed biofilm reactors.

    PubMed

    Jaroszynski, L W; Cicek, N; Sparling, R; Oleszkiewicz, J A

    2011-07-01

    Two bench-scale parallel moving bed biofilm reactors (MBBR) were operated to assess pH-associated anammox activity changes during long term treatment of anaerobically digested sludge centrate pre-treated in a suspended growth partial nitrification reactor. The pH was maintained at 6.5 in reactor R1, while it was allowed to vary naturally between 7.5 and 8.1 in reactor R2. At high nitrogen loads reactor R2 had a 61% lower volumetric specific nitrogen removal rate than reactor R1. The low pH and the associated low free ammonia (FA) concentrations were found to be critical to stable anammox activity in the MBBR. Nitrite enhanced the nitrogen removal rate in the conditions of low pH, all the way up to the investigated level of 50mg NO(2)-N/L. At low FA levels nitrite concentrations up to 250 mg NO(2)-N/L did not cause inactivation of anammox consortia over a 2-days exposure time.

  15. Human Erythropoietin Dimers with Markedly Enhanced in vivo Activity

    NASA Astrophysics Data System (ADS)

    Sytkowski, Arthur J.; Dotimas Lunn, Elizabeth; Davis, Kerry Lynn; Feldman, Laurie; Siekman, Suvia

    1998-02-01

    Human erythropoietin, a widely used and important therapeutic glycoprotein, has a relatively short plasma half-life due to clearance by glomerular filtration as well as by other mechanisms. We hypothesized that an erythropoietin species with a larger molecular size would exhibit an increased plasma half-life and, potentially, an enhanced biological activity. We now report the production of biologically active erythropoietin dimers and trimers by chemical crosslinking of the conventional monomeric form. We imparted free sulfhydryl residues to a pool of erythropoietin monomer by chemical modification. A second pool was reacted with another modifying reagent to yield monomer with male-imido groups. Upon mixing these two pools, covalently linked dimers and trimers were formed that were biologically active in vitro. The plasma half-life of erythropoietin dimers in rabbits was >24 h compared with 4 h for the monomers. Importantly, erythropoietin dimers were biologically active in vivo as shown by their ability to increase the hematocrits of mice when injected subcutaneously. In addition, the dimers exhibited >26-fold higher activity in vivo than did the monomers and were very effective after only one dose. Dimeric and other oligomeric forms of Epo may have an important role in therapy.

  16. The importance of active surveillance, and immediate re-biopsy in low-risk prostate cancer: The largest series from Turkey

    PubMed Central

    Bayar, Göksel; Horasanlı, Kaya; Acinikli, Hüseyin; Tanrıverdi, Orhan; Dalkılıç, Ayhan; Arısan, Serdar

    2016-01-01

    Objective To evaluate long-term outcomes of active surveillance (AS) applied in low-risk prostate cancer patients, and the impact of re-biopsy results on the prediction of progression. Material and methods In our clinic, patients who had undergone AS for low-risk localized prostate cancer between the years 2005–2013 were included in the study. Our AS criteria are Gleason score ≤6, prostate-specific antigen (PSA) level <10 ng/mL, number of positive cores <3, maximum cancer involvement ratio <50% each core. Immediate re-biopsy (within 3 months) was performed to 65 patients who accepted AS. Finally, 43 patients who met re-biopsy criteria were included in the study. Prostate biopsy specimens were harvested from 12 cores under the guidance of transrectal ultrasound (TRUS). Re-biopsy was performed within 3 months (1–12 weeks). In re-biopsy, a total of 20 core biopsies were performed including the far lateral (6 cores) and transition zone (2 cores) in addition to standard 12 core biopsy. Our follow-up protocol is PSA measurement and digital rectal examination (DRE) every 3 months within the first 2 years, than every 6 months. Control biopsies was performed one year later and once upon every 3 years to patients whose PSA levels and DREs were normal at follow-up visits. More than 2 tumor invaded cores or 50% tumor in one core, and Gleason score exceeding 6 points were accepted as indications for definitive treatment. Patients were divided into two groups by re-biopsy results and compared according to the time to progression. We have done multivariate regression analysis to predict prognosis by using data on age, PSA level, and detection of tumor in re-biopsy specimens. Results Patients’ median age was 61 years and PSA level was 5 (2.7–9) ng/mL. Tumor was detected in 22 (34%) patients at re-biopsy and they underwent definitive treatment. Additionally tumor was detected in 9 patients, but active surveillance was maintained because their pathologic results met active

  17. Structure-activity relationship of genotoxic polycyclic aromatic nitro compounds: Further evidence for the importance of hydrophobicity and molecular orbital energies in genetic toxicity

    SciTech Connect

    Debnath, A.K.; Hansch, C. )

    1992-01-01

    A quantitative structure-activity relationship (QSAR) has been formulated for 15 polycyclic aromatic nitro compounds acting on E. coli PQ37. Upon damage of DNA by these substances [beta]-galactosidase is induced and can be easily assayed colorimetrically, hence, this is a short-term test for mutagenicity. The QSAR (log SOSIP = 1.07 log P - 1.57 e[sub LUMO] - 6.41) is strikingly similar to that found earlier with nitroaromatics acting in the Ames test (TA100) and differs significantly for that found using TA98 organisms. The QSAR brings out in a unique manner the underlying similarity in the two test systems. 24 refs., 2 tabs.

  18. The class II HD-ZIP JAIBA gene is involved in meristematic activity and important for gynoecium and fruit development in Arabidopsis

    PubMed Central

    Zúñiga-Mayo, Victor M.; Marsch-Martínez, Nayelli; de Folter, Stefan

    2012-01-01

    Development and patterning of the gynoecium – and later the fruit – must be finely regulated to ensure the survival of the species that produces them. The process that leads to successful fruit formation starts at early stages of floral meristem development and follows a series of chronologically successive events. In a recent work we reported the functional characterization of the class II HD-ZIP JAIBA (JAB) gene. Mutant jab plants showed sporophytic defects in male and female reproductive development, and combined with the mutant crabs claw (crc) caused defects in the floral meristem (FM) determination process and gynoecium medial tissue development. Furthermore, the JAB protein interacted with transcription factors known to regulate meristematic activity, fruit development and FM determinacy. Preliminary results presented here suggest a genetic interaction between JAB and the gene SHOOT MERISTEMLESS (STM). PMID:22951401

  19. The class II HD-ZIP JAIBA gene is involved in meristematic activity and important for gynoecium and fruit development in Arabidopsis.

    PubMed

    Zúñiga-Mayo, Victor M; Marsch-Martínez, Nayelli; de Folter, Stefan

    2012-11-01

    Development and patterning of the gynoecium - and later the fruit - must be finely regulated to ensure the survival of the species that produces them. The process that leads to successful fruit formation starts at early stages of floral meristem development and follows a series of chronologically successive events. In a recent work we reported the functional characterization of the class II HD-ZIP JAIBA (JAB) gene. Mutant jab plants showed sporophytic defects in male and female reproductive development, and combined with the mutant crabs claw (crc) caused defects in the floral meristem (FM) determination process and gynoecium medial tissue development. Furthermore, the JAB protein interacted with transcription factors known to regulate meristematic activity, fruit development and FM determinacy. Preliminary results presented here suggest a genetic interaction between JAB and the gene SHOOT MERISTEMLESS (STM). PMID:22951401

  20. Importance of penicillinase production for activity of penicillin alone or in combination with sulbactam in experimental endocarditis due to methicillin-resistant Staphylococcus aureus.

    PubMed Central

    Fantin, B; Pierre, J; Castéla-Papin, N; Saint-Julien, L; Drugeon, H; Farinotti, R; Carbon, C

    1996-01-01

    The activity of penicillin, alone and in combination with sulbactam, against a heterogeneously methicillin-resistant, penicillinase-producing clinical isolate of Staphylococcus aureus and its penicillinase-negative derivative was investigated in vitro and in a rabbit experimental endocarditis model. Penicillin was significantly more effective than vancomycin against the penicillinase-negative derivative in vivo (P < 0.001), and it sterilized 25% of the vegetations. The combination of penicillin and sulbactam exhibited an in vivo synergistic effect on the penicillinase-producing strain (P < 0.01) but did not produce any advantage over treatment with vancomycin, even when a high dose of sulbactam was used (100 mg/kg of body weight every 6 h). This combination was significantly less effective against the penicillinase-producing strain than was penicillin alone against the penicillinase-negative derivative (P < 0.03). In addition, the most resistant subpopulation of the surviving bacteria, which grew on agar containing 16 micrograms of methicillin per ml, was detected in 5 of 6 animals treated with penicillin and a high dose of sulbactam against the penicillinase-producing strain compared with only 1 of 12 animals treated with penicillin alone against the penicillinase-negative derivative (P < 0.01). We conclude that penicillin is highly effective against penicillinase-negative methicillin-resistant S. aureus and that penicillinase production, rather than methicillin resistance, appears to be the limiting factor for the activity of the penicillin-sulbactam combination against penicillinase-producing, methicillin-resistant S. aureus. PMID:8723470

  1. Anti-nociceptive Activity of Ethnomedicinally Important Analgesic Plant Isodon rugosus Wall. ex Benth: Mechanistic Study and Identifications of Bioactive Compounds

    PubMed Central

    Zeb, Anwar; Ahmad, Sajjad; Ullah, Farhat; Ayaz, Muhammad; Sadiq, Abdul

    2016-01-01

    Isodon rugosus Wall. ex Benth. is extensively used as traditional medicine for the management of various types of pain including tooth ache, gastric pain, abdominal pain, ear ache, and generalized body pain. The current study is designed to scientifically verify the purported uses of I. rugosus as analgesic agent and to figure out its possible mechanism of action. Bioactive compounds responsible for analgesic activity were identified using GC and GC-MS analysis. Analgesic potentials were evaluated using acetic acid induced writhing, hot plate test, and formalin induced paw licking test. In acetic acid induced writhing chloroform fraction (Ir.Chf) exhibited 53% analgesia while formalin test displayed 61% inhibition at phase-I and 45% at phase-II respectively at a dose of 100 mg/kg. Similarly, in hot plate test Ir.Chf displayed average reaction time of 7 min at 15, 30, 45, and 60 min intervals. The possible mechanism of action was found to be the central pathway via opioidergic receptors as the mice showed morphine like analgesic activity at pre-administration of naloxone (opioid antagonist) in hot plate and formalin tests. In GC-MS analysis, 83 compounds were identified among which eight compounds including benzyl alcohol, sebacic acid, myristic acid, phytol, sugiol, Tocopherol, α-Amyrin, and stigmasterol were sorted out as previously reported analgesic compounds. Current study revealed that analgesic potential of I. rugosus can attributed to the presence of analgesic compounds. It may also be concluded that opioids receptors are involved in the analgesic mechanism of I. rugosus due to effective antagonism of nalaxone. PMID:27458379

  2. Familial Risk of Chronic Musculoskeletal Pain and the Importance of Physical Activity and Body Mass Index: Prospective Data from the HUNT Study, Norway.

    PubMed

    Lier, Ragnhild; Mork, Paul Jarle; Holtermann, Andreas; Nilsen, Tom Ivar Lund

    2016-01-01

    The main objectives of the current study was i) to prospectively examine if chronic musculoskeletal pain in parents is associated with risk of chronic musculoskeletal pain in their adult offspring, and ii) to assess if these parent-offspring associations are modified by offspring body mass index and leisure time physical activity. We used data on 4,742 adult offspring linked with their parents who participated in the population-based HUNT Study in Norway in 1995-97 and in 2006-08. Family relations were established through the national Family Registry. A Poisson regression model was used to estimate relative risk (RR) with 95% confidence interval (CI). In total, 1,674 offspring (35.3%) developed chronic musculoskeletal pain during the follow-up period of approximately 11 years. Both maternal (RR: 1.26, 95% CI: 1.03, 1.55) and paternal chronic musculoskeletal pain (RR: 1.29, 95% CI: 1.06, 1.57) was associated with increased risk of offspring chronic musculoskeletal pain. Compared to offspring of parents without chronic musculoskeletal pain, the adverse effect of parental pain was somewhat stronger among offspring who reported a low (RR: 1.82, 95% CI: 1.32, 2.52) versus high (RR: 1.32, 95% CI: 0.95, 1.84) level of leisure time physical activity. Offspring of parents with chronic musculoskeletal pain and who were classified as obese had more than twofold increased risk (RR: 2.33, 95% CI: 1.68, 3.24) of chronic musculoskeletal pain compared to normal weight offspring of parents without pain. In conclusion, parental chronic musculoskeletal pain is positively associated with risk of chronic musculoskeletal pain in their adult offspring. Maintenance of normal body weight may reduce the risk of chronic musculoskeletal pain in offspring of pain-afflicted parents.

  3. Anti-nociceptive Activity of Ethnomedicinally Important Analgesic Plant Isodon rugosus Wall. ex Benth: Mechanistic Study and Identifications of Bioactive Compounds.

    PubMed

    Zeb, Anwar; Ahmad, Sajjad; Ullah, Farhat; Ayaz, Muhammad; Sadiq, Abdul

    2016-01-01

    Isodon rugosus Wall. ex Benth. is extensively used as traditional medicine for the management of various types of pain including tooth ache, gastric pain, abdominal pain, ear ache, and generalized body pain. The current study is designed to scientifically verify the purported uses of I. rugosus as analgesic agent and to figure out its possible mechanism of action. Bioactive compounds responsible for analgesic activity were identified using GC and GC-MS analysis. Analgesic potentials were evaluated using acetic acid induced writhing, hot plate test, and formalin induced paw licking test. In acetic acid induced writhing chloroform fraction (Ir.Chf) exhibited 53% analgesia while formalin test displayed 61% inhibition at phase-I and 45% at phase-II respectively at a dose of 100 mg/kg. Similarly, in hot plate test Ir.Chf displayed average reaction time of 7 min at 15, 30, 45, and 60 min intervals. The possible mechanism of action was found to be the central pathway via opioidergic receptors as the mice showed morphine like analgesic activity at pre-administration of naloxone (opioid antagonist) in hot plate and formalin tests. In GC-MS analysis, 83 compounds were identified among which eight compounds including benzyl alcohol, sebacic acid, myristic acid, phytol, sugiol, Tocopherol, α-Amyrin, and stigmasterol were sorted out as previously reported analgesic compounds. Current study revealed that analgesic potential of I. rugosus can attributed to the presence of analgesic compounds. It may also be concluded that opioids receptors are involved in the analgesic mechanism of I. rugosus due to effective antagonism of nalaxone. PMID:27458379

  4. Familial Risk of Chronic Musculoskeletal Pain and the Importance of Physical Activity and Body Mass Index: Prospective Data from the HUNT Study, Norway.

    PubMed

    Lier, Ragnhild; Mork, Paul Jarle; Holtermann, Andreas; Nilsen, Tom Ivar Lund

    2016-01-01

    The main objectives of the current study was i) to prospectively examine if chronic musculoskeletal pain in parents is associated with risk of chronic musculoskeletal pain in their adult offspring, and ii) to assess if these parent-offspring associations are modified by offspring body mass index and leisure time physical activity. We used data on 4,742 adult offspring linked with their parents who participated in the population-based HUNT Study in Norway in 1995-97 and in 2006-08. Family relations were established through the national Family Registry. A Poisson regression model was used to estimate relative risk (RR) with 95% confidence interval (CI). In total, 1,674 offspring (35.3%) developed chronic musculoskeletal pain during the follow-up period of approximately 11 years. Both maternal (RR: 1.26, 95% CI: 1.03, 1.55) and paternal chronic musculoskeletal pain (RR: 1.29, 95% CI: 1.06, 1.57) was associated with increased risk of offspring chronic musculoskeletal pain. Compared to offspring of parents without chronic musculoskeletal pain, the adverse effect of parental pain was somewhat stronger among offspring who reported a low (RR: 1.82, 95% CI: 1.32, 2.52) versus high (RR: 1.32, 95% CI: 0.95, 1.84) level of leisure time physical activity. Offspring of parents with chronic musculoskeletal pain and who were classified as obese had more than twofold increased risk (RR: 2.33, 95% CI: 1.68, 3.24) of chronic musculoskeletal pain compared to normal weight offspring of parents without pain. In conclusion, parental chronic musculoskeletal pain is positively associated with risk of chronic musculoskeletal pain in their adult offspring. Maintenance of normal body weight may reduce the risk of chronic musculoskeletal pain in offspring of pain-afflicted parents. PMID:27082110

  5. Familial Risk of Chronic Musculoskeletal Pain and the Importance of Physical Activity and Body Mass Index: Prospective Data from the HUNT Study, Norway

    PubMed Central

    Lier, Ragnhild; Mork, Paul Jarle; Holtermann, Andreas; Nilsen, Tom Ivar Lund

    2016-01-01

    The main objectives of the current study was i) to prospectively examine if chronic musculoskeletal pain in parents is associated with risk of chronic musculoskeletal pain in their adult offspring, and ii) to assess if these parent-offspring associations are modified by offspring body mass index and leisure time physical activity. We used data on 4,742 adult offspring linked with their parents who participated in the population-based HUNT Study in Norway in 1995–97 and in 2006–08. Family relations were established through the national Family Registry. A Poisson regression model was used to estimate relative risk (RR) with 95% confidence interval (CI). In total, 1,674 offspring (35.3%) developed chronic musculoskeletal pain during the follow-up period of approximately 11 years. Both maternal (RR: 1.26, 95% CI: 1.03, 1.55) and paternal chronic musculoskeletal pain (RR: 1.29, 95% CI: 1.06, 1.57) was associated with increased risk of offspring chronic musculoskeletal pain. Compared to offspring of parents without chronic musculoskeletal pain, the adverse effect of parental pain was somewhat stronger among offspring who reported a low (RR: 1.82, 95% CI: 1.32, 2.52) versus high (RR: 1.32, 95% CI: 0.95, 1.84) level of leisure time physical activity. Offspring of parents with chronic musculoskeletal pain and who were classified as obese had more than twofold increased risk (RR: 2.33, 95% CI: 1.68, 3.24) of chronic musculoskeletal pain compared to normal weight offspring of parents without pain. In conclusion, parental chronic musculoskeletal pain is positively associated with risk of chronic musculoskeletal pain in their adult offspring. Maintenance of normal body weight may reduce the risk of chronic musculoskeletal pain in offspring of pain-afflicted parents. PMID:27082110

  6. The importance of active surveillance, and immediate re-biopsy in low-risk prostate cancer: The largest series from Turkey

    PubMed Central

    Bayar, Göksel; Horasanlı, Kaya; Acinikli, Hüseyin; Tanrıverdi, Orhan; Dalkılıç, Ayhan; Arısan, Serdar

    2016-01-01

    Objective To evaluate long-term outcomes of active surveillance (AS) applied in low-risk prostate cancer patients, and the impact of re-biopsy results on the prediction of progression. Material and methods In our clinic, patients who had undergone AS for low-risk localized prostate cancer between the years 2005–2013 were included in the study. Our AS criteria are Gleason score ≤6, prostate-specific antigen (PSA) level <10 ng/mL, number of positive cores <3, maximum cancer involvement ratio <50% each core. Immediate re-biopsy (within 3 months) was performed to 65 patients who accepted AS. Finally, 43 patients who met re-biopsy criteria were included in the study. Prostate biopsy specimens were harvested from 12 cores under the guidance of transrectal ultrasound (TRUS). Re-biopsy was performed within 3 months (1–12 weeks). In re-biopsy, a total of 20 core biopsies were performed including the far lateral (6 cores) and transition zone (2 cores) in addition to standard 12 core biopsy. Our follow-up protocol is PSA measurement and digital rectal examination (DRE) every 3 months within the first 2 years, than every 6 months. Control biopsies was performed one year later and once upon every 3 years to patients whose PSA levels and DREs were normal at follow-up visits. More than 2 tumor invaded cores or 50% tumor in one core, and Gleason score exceeding 6 points were accepted as indications for definitive treatment. Patients were divided into two groups by re-biopsy results and compared according to the time to progression. We have done multivariate regression analysis to predict prognosis by using data on age, PSA level, and detection of tumor in re-biopsy specimens. Results Patients’ median age was 61 years and PSA level was 5 (2.7–9) ng/mL. Tumor was detected in 22 (34%) patients at re-biopsy and they underwent definitive treatment. Additionally tumor was detected in 9 patients, but active surveillance was maintained because their pathologic results met active

  7. BdorCSP2 Is Important for Antifeed and Oviposition-Deterring Activities Induced by Rhodojaponin-III against Bactrocera dorsalis

    PubMed Central

    Dong, Xiaolin; Wang, Peidan; Hu, Meiying; Zhong, Guohua

    2013-01-01

    Rhodojaponin-III is a nonvolatile botanical grayanoid diterpene compound, which has antifeedant and oviposition deterrence effects against many kinds of insects. However, the molecular mechanism of the chemoreception process remains unknown. In this study, the important role of BdorCSP2 in the recognition of Rhodojaponin-III was identified. The full length cDNA encoding BdorCSP2 was cloned from legs of Bactrocera dorsalis. The results of expression pattern revealed that BdorCSP2 was abundantly expressed in the legs of adult B. dorsalis. Moreover, the expression of BdorCSP2 could be up-regulated by Rhodojaponin-III. In order to gain comprehensive understanding of the recognition process, the binding affinity between BdorCSP2 and Rhodojaponin-III was measured by fluorescence binding assay. Silencing the expression of BdorCSP2 through the ingestion of dsRNA could weaken the effect of oviposition deterrence and antifeedant of Rhodojaponin-III. These results suggested that BdorCSP2 of B. dorsalis could be involved in chemoreception of Rhodojaponin-III and played a critical role in antifeedant and oviposition behaviors induced by Rhodojaponin-III. PMID:24155937

  8. Post-thaw viability of cryopreserved peripheral blood stem cells (PBSC) does not guarantee functional activity: important implications for quality assurance of stem cell transplant programmes.

    PubMed

    Morgenstern, Daniel A; Ahsan, Gulrukh; Brocklesby, Margaret; Ings, Stuart; Balsa, Carmen; Veys, Paul; Brock, Penelope; Anderson, John; Amrolia, Persis; Goulden, Nicholas; Cale, Catherine M; Watts, Michael J

    2016-09-01

    Standard quality assurance (QA) of cryopreserved peripheral blood stem cells (PBSC) uses post-thaw viable CD34(+) cell counts. In 2013, concerns arose at Great Ormond Street Hospital (GOSH) about 8 patients with delayed engraftment following myeloablative chemotherapy with cryopreserved cell rescue, despite adequate post-thaw viable cell counts in all cases. Root cause analysis was undertaken; investigations suggested the freeze process itself was a contributing factor to suboptimal engraftment. Experiments were undertaken in which a single PBSC product was divided into three and cryopreserved in parallel using a control-rate freezer (CRF) or passive freezing method (-80°C freezer) at GOSH, or the same passive freezing at another laboratory. Viable CD34(+) counts were equivalent and adequate in each. Granulocyte-monocyte colony-forming unit assays demonstrated colonies from the products cryopreserved using passive freezing (both laboratories), but no colonies from products cryopreserved using the CRF. The CRF was shown to be operating within manufacturer's specifications with freeze-profile within acceptable limits. This experience has important implications for quality assurance for all transplant programmes, particularly those using cryopreserved products. The failure of post-thaw viable CD34(+) counts, the most widely used routine QA test available, to ensure PBSC function is of great concern and should prompt reassessment of protocols and QA procedures. PMID:27291859

  9. Post-thaw viability of cryopreserved peripheral blood stem cells (PBSC) does not guarantee functional activity: important implications for quality assurance of stem cell transplant programmes.

    PubMed

    Morgenstern, Daniel A; Ahsan, Gulrukh; Brocklesby, Margaret; Ings, Stuart; Balsa, Carmen; Veys, Paul; Brock, Penelope; Anderson, John; Amrolia, Persis; Goulden, Nicholas; Cale, Catherine M; Watts, Michael J

    2016-09-01

    Standard quality assurance (QA) of cryopreserved peripheral blood stem cells (PBSC) uses post-thaw viable CD34(+) cell counts. In 2013, concerns arose at Great Ormond Street Hospital (GOSH) about 8 patients with delayed engraftment following myeloablative chemotherapy with cryopreserved cell rescue, despite adequate post-thaw viable cell counts in all cases. Root cause analysis was undertaken; investigations suggested the freeze process itself was a contributing factor to suboptimal engraftment. Experiments were undertaken in which a single PBSC product was divided into three and cryopreserved in parallel using a control-rate freezer (CRF) or passive freezing method (-80°C freezer) at GOSH, or the same passive freezing at another laboratory. Viable CD34(+) counts were equivalent and adequate in each. Granulocyte-monocyte colony-forming unit assays demonstrated colonies from the products cryopreserved using passive freezing (both laboratories), but no colonies from products cryopreserved using the CRF. The CRF was shown to be operating within manufacturer's specifications with freeze-profile within acceptable limits. This experience has important implications for quality assurance for all transplant programmes, particularly those using cryopreserved products. The failure of post-thaw viable CD34(+) counts, the most widely used routine QA test available, to ensure PBSC function is of great concern and should prompt reassessment of protocols and QA procedures.

  10. Characterization of the Mycobacterium tuberculosis H37Rv alkyl hydroperoxidase AhpC points to the importance of ionic interactions in oligomerization and activity.

    PubMed Central

    Chauhan, R; Mande, S C

    2001-01-01

    An alkyl hydroperoxidase (AhpC) has been found frequently to be overexpressed in isoniazid-resistant strains of Mycobacterium tuberculosis. These strains have an inactivated katG gene encoding a catalase peroxidase, which might render mycobacteria susceptible to the toxic peroxide radicals, thus leading to the concomitant overexpression of the AhpC. Although the overexpressed AhpC in isoniazid-resistant strains of M. tuberculosis may not directly participate in isoniazid action, AhpC might still assist M. tuberculosis in combating oxidative damage in the absence of the catalase. Here we have attempted to characterize the AhpC protein biochemically and report its functional and oligomerization properties. The alkyl hydroperoxidase of M. tuberculosis is unique in many ways compared with its well-characterized homologues from enteric bacteria. We show that AhpC is a decameric protein, composed of five identical dimers held together by ionic interactions. Dimerization of individual subunits takes place through an intersubunit disulphide linkage. The ionic interactions play a significant role in enzymic activity of the AhpC protein. The UV absorption spectrum and three-dimensional model of AhpC suggest that interesting conformational changes may take place during oxidation and reduction of the intersubunit disulphide linkage. In the absence of the partner AhpF subunit in M. tuberculosis, the mycobacterial AhpC might use small-molecule reagents, such as mycothiol, for completing its enzymic cycle. PMID:11171096

  11. Lnk is an important modulator of insulin-like growth factor-1/Akt/peroxisome proliferator-activated receptor-gamma axis during adipogenesis of mesenchymal stem cells

    PubMed Central

    Lee, Jun Hee; Lee, Sang Hun; Lee, Hyang Seon; Ji, Seung Taek; Jung, Seok Yun; Kim, Jae Ho; Bae, Sun Sik

    2016-01-01

    Adipogenic differentiation of mesenchymal stem cells (MSCs) is critical for metabolic homeostasis and nutrient signaling during development. However, limited information is available on the pivotal modulators of adipogenic differentiation of MSCs. Adaptor protein Lnk (Src homology 2B3 [SH2B3]), which belongs to a family of SH2-containing proteins, modulates the bioactivities of different stem cells, including hematopoietic stem cells and endothelial progenitor cells. In this study, we investigated whether an interaction between insulin-like growth factor-1 receptor (IGF-1R) and Lnk regulated IGF-1-induced adipogenic differentiation of MSCs. We found that wild-type MSCs showed greater adipogenic differentiation potential than Lnk–/– MSCs. An ex vivo adipogenic differentiation assay showed that Lnk–/– MSCs had decreased adipogenic differentiation potential compared with wild-type MSCs. Interestingly, we found that Lnk formed a complex with IGF-1R and that IGF-1 induced the dissociation of this complex. In addition, we observed that IGF-1-induced increase in the phosphorylation of Akt and mammalian target of rapamycin was triggered by the dissociation of the IGF-1R–Lnk complex. Expression levels of a pivotal transcription factor peroxisome proliferator-activated receptor gamma (PPAR-γ) and its adipogenic target genes (LPL and FABP4) significantly decreased in Lnk–/– MSCs. These results suggested that Lnk adaptor protein regulated the adipogenesis of MSCs through the IGF-1/Akt/PPAR-γ pathway. PMID:27610032

  12. Lnk is an important modulator of insulin-like growth factor-1/Akt/peroxisome proliferator-activated receptor-gamma axis during adipogenesis of mesenchymal stem cells.

    PubMed

    Lee, Jun Hee; Lee, Sang Hun; Lee, Hyang Seon; Ji, Seung Taek; Jung, Seok Yun; Kim, Jae Ho; Bae, Sun Sik; Kwon, Sang-Mo

    2016-09-01

    Adipogenic differentiation of mesenchymal stem cells (MSCs) is critical for metabolic homeostasis and nutrient signaling during development. However, limited information is available on the pivotal modulators of adipogenic differentiation of MSCs. Adaptor protein Lnk (Src homology 2B3 [SH2B3]), which belongs to a family of SH2-containing proteins, modulates the bioactivities of different stem cells, including hematopoietic stem cells and endothelial progenitor cells. In this study, we investigated whether an interaction between insulin-like growth factor-1 receptor (IGF-1R) and Lnk regulated IGF-1-induced adipogenic differentiation of MSCs. We found that wild-type MSCs showed greater adipogenic differentiation potential than Lnk (-/-) MSCs. An ex vivo adipogenic differentiation assay showed that Lnk (-/-) MSCs had decreased adipogenic differentiation potential compared with wild-type MSCs. Interestingly, we found that Lnk formed a complex with IGF-1R and that IGF-1 induced the dissociation of this complex. In addition, we observed that IGF-1-induced increase in the phosphorylation of Akt and mammalian target of rapamycin was triggered by the dissociation of the IGF-1R-Lnk complex. Expression levels of a pivotal transcription factor peroxisome proliferator-activated receptor gamma (PPAR-γ) and its adipogenic target genes (LPL and FABP4) significantly decreased in Lnk (-/-) MSCs. These results suggested that Lnk adaptor protein regulated the adipogenesis of MSCs through the IGF-1/Akt/PPAR-γ pathway. PMID:27610032

  13. Juglans regia and J. nigra, two trees important in traditional medicine: A comparison of leaf essential oil compositions and biological activities.

    PubMed

    Paudel, Prajwal; Satyal, Prabodh; Dosoky, Noura S; Maharjan, Samjhana; Setzer, William N

    2013-10-01

    The essential oils from the leaves of Juglans regia, collected from Kathmandu, Nepal, and Juglans nigra from Huntsville, Alabama, USA, were obtained by hydrodistillation and analyzed by GC-MS. A total of 69 compounds were identified in the J. regia oil, accounting for 99.0% of the eluting components of the oil while three different samples of J. nigra were analyzed identifying a total of 99.3%, 99.4% and 99.7% of the oils. The leaf essential oil of J. regia was dominated by the aromatic compounds eugenol (27.5%) and methyl salicylate (16.2%), and the sesquiterpenes germacrene D (21.4%) and (E)-beta-farnesene (8.2%). The essential oils from three different samples of. J. nigra contained (E)-caryophyllene (17.3%-20.4%) and germacrene D (7.1%-22.5%) with smaller amounts of juglone (1.0%-8.8%), alpha-hydrojuglone (1.0-9.5%), and delta-cadinene (3.8%-8.7%). J. regia leaf oil, devoid of juglone, showed allelopathic activity, while J. nigra leaf oil was less phytotoxic. The presence of eugenol and methyl salicylate in the leaves of J. regia are consistent with the traditional uses of this plant to treat toothache, rheumatism, and fungal infections.

  14. Lnk is an important modulator of insulin-like growth factor-1/Akt/peroxisome proliferator-activated receptor-gamma axis during adipogenesis of mesenchymal stem cells

    PubMed Central

    Lee, Jun Hee; Lee, Sang Hun; Lee, Hyang Seon; Ji, Seung Taek; Jung, Seok Yun; Kim, Jae Ho; Bae, Sun Sik

    2016-01-01

    Adipogenic differentiation of mesenchymal stem cells (MSCs) is critical for metabolic homeostasis and nutrient signaling during development. However, limited information is available on the pivotal modulators of adipogenic differentiation of MSCs. Adaptor protein Lnk (Src homology 2B3 [SH2B3]), which belongs to a family of SH2-containing proteins, modulates the bioactivities of different stem cells, including hematopoietic stem cells and endothelial progenitor cells. In this study, we investigated whether an interaction between insulin-like growth f