Sample records for question alternative oxidase

  1. Expression of alternative oxidase in tomato

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    Kakefuda, M.; McIntosh, L. (Michigan State Univ., East Lansing (USA))


    Tomato fruit ripening is characterized by an increase in ethylene biosynthesis, a burst in respiration (i.e. the climacteric), fruit softening and pigmentation. As whole tomatoes ripened from mature green to red, there was an increase in the alternative oxidase capacity. Aging pink tomato slices for 24 and 48 hrs also showed an increase of alternative oxidase and cytochrome oxidase capacities. Monoclonal antibodies prepared to the Sauromatum guttatum alternative oxidase were used to follow the appearance of alternative oxidase in tomato fruits. There is a corresponding increase in a 36kDa protein with an increase in alternative oxidase capacity. Effects of ethylene and norbornadiene on alternative oxidase capacity were also studied. We are using an alternative oxidase cDNA clone from potato to study the expression of mRNA in ripening and wounded tomatoes to determine if the gene is transcriptionally regulated.

  2. Alternative oxidase: distribution, induction, properties, structure, regulation, and functions. (United States)

    Rogov, A G; Sukhanova, E I; Uralskaya, L A; Aliverdieva, D A; Zvyagilskaya, R A


    The respiratory chain in the majority of organisms with aerobic type metabolism features the concomitant existence of the phosphorylating cytochrome pathway and the cyanide- and antimycin A-insensitive oxidative route comprising a so-called alternative oxidase (AOX) as a terminal oxidase. In this review, the history of AOX discovery is described. Considerable evidence is presented that AOX occurs widely in organisms at various levels of organization and is not confined to the plant kingdom. This enzyme has not been found only in Archaea, mammals, some yeasts and protists. Bioinformatics research revealed the sequences characteristic of AOX in representatives of various taxonomic groups. Based on multiple alignments of these sequences, a phylogenetic tree was constructed to infer their possible evolution. The ways of AOX activation, as well as regulatory interactions between AOX and the main respiratory chain are described. Data are summarized concerning the properties of AOX and the AOX-encoding genes whose expression is either constitutive or induced by various factors. Information is presented on the structure of AOX, its active center, and the ubiquinone-binding site. The principal functions of AOX are analyzed, including the cases of cell survival, optimization of respiratory metabolism, protection against excess of reactive oxygen species, and adaptation to variable nutrition sources and to biotic and abiotic stress factors. It is emphasized that different AOX functions complement each other in many instances and are not mutually exclusive. Examples are given to demonstrate that AOX is an important tool to overcome the adverse aftereffects of restricted activity of the main respiratory chain in cells and whole animals. This is the first comprehensive review on alternative oxidases of various organisms ranging from yeasts and protists to vascular plants.

  3. A broad distribution of the alternative oxidase in microsporidian parasites.

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    Bryony A P Williams


    Full Text Available Microsporidia are a group of obligate intracellular parasitic eukaryotes that were considered to be amitochondriate until the recent discovery of highly reduced mitochondrial organelles called mitosomes. Analysis of the complete genome of Encephalitozoon cuniculi revealed a highly reduced set of proteins in the organelle, mostly related to the assembly of iron-sulphur clusters. Oxidative phosphorylation and the Krebs cycle proteins were absent, in keeping with the notion that the microsporidia and their mitosomes are anaerobic, as is the case for other mitosome bearing eukaryotes, such as Giardia. Here we provide evidence opening the possibility that mitosomes in a number of microsporidian lineages are not completely anaerobic. Specifically, we have identified and characterized a gene encoding the alternative oxidase (AOX, a typically mitochondrial terminal oxidase in eukaryotes, in the genomes of several distantly related microsporidian species, even though this gene is absent from the complete genome of E. cuniculi. In order to confirm that these genes encode functional proteins, AOX genes from both A. locustae and T. hominis were over-expressed in E. coli and AOX activity measured spectrophotometrically using ubiquinol-1 (UQ-1 as substrate. Both A. locustae and T. hominis AOX proteins reduced UQ-1 in a cyanide and antimycin-resistant manner that was sensitive to ascofuranone, a potent inhibitor of the trypanosomal AOX. The physiological role of AOX microsporidia may be to reoxidise reducing equivalents produced by glycolysis, in a manner comparable to that observed in trypanosomes.

  4. Questions over alternatives to detention programmes

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    Stephanie J Silverman


    Full Text Available Alternative to detention programmes may be less restrictive and less expensive than formal detention but they may still have drawbacks. The provision of competent legal advice appears to be key to low rates of absconding.

  5. Supramolecular organization of cytochrome c oxidase- and alternative oxidase-dependent respiratory chains in the filamentous fungus Podospora anserina. (United States)

    Krause, Frank; Scheckhuber, Christian Q; Werner, Alexandra; Rexroth, Sascha; Reifschneider, Nicole H; Dencher, Norbert A; Osiewacz, Heinz D


    To elucidate the molecular basis of the link between respiration and longevity, we have studied the organization of the respiratory chain of a wild-type strain and of two long-lived mutants of the filamentous fungus Podospora anserina. This established aging model is able to respire by either the standard or the alternative pathway. In the latter pathway, electrons are directly transferred from ubiquinol to the alternative oxidase and thus bypass complexes III and IV. We show that the cytochrome c oxidase pathway is organized according to the mammalian "respirasome" model (Schägger, H., and Pfeiffer, K. (2000) EMBO J. 19, 1777-1783). In contrast, the alternative pathway is composed of distinct supercomplexes of complexes I and III (i.e. I(2) and I(2)III(2)), which have not been described so far. Enzymatic analysis reveals distinct functional properties of complexes I and III belonging to either cytochrome c oxidase- or alternative oxidase-dependent pathways. By a gentle colorless-native PAGE, almost all of the ATP synthases from mitochondria respiring by either pathway were preserved in the dimeric state. Our data are of significance for the understanding of both respiratory pathways as well as lifespan control and aging.

  6. Alternative oxidase involvement in Daucus carota somatic embryogenesis. (United States)

    Frederico, António Miguel; Campos, Maria Doroteia; Cardoso, Hélia Guerra; Imani, Jafargholi; Arnholdt-Schmitt, Birgit


    Plant alternative oxidase (AOX) is a mitochondrial inner membrane enzyme involved in alternative respiration. The critical importance of the enzyme during acclimation upon stress of plant cells is not fully understood and is still an issue of intensive research and discussion. Recently, a role of AOX was suggested for the ability of plant cells to change easily its fate upon stress. In order to get new insights about AOX involvement in cell reprogramming, quantitative real-time polymerase chain reaction (PCR) and inhibitor studies were performed during cell redifferentiation and developmental stages of Daucus carota L. somatic embryogenesis. Transcript level analysis shows that D. carota AOX genes (DcAOX1a and DcAOX2a) are differentially expressed during somatic embryogenesis. DcAOX1a shows lower expression levels, being mainly down-regulated, whereas DcAOX2a presented a large up-regulation during initiation of the realization phase of somatic embryogenesis. However, when globular embryos start to develop, both genes are down-regulated, being this state transient for DcAOX2a. In addition, parallel studies were performed using salicylhydroxamic acid (SHAM) in order to inhibit AOX activity during the realization phase of somatic embryogenesis. Embryogenic cells growing in the presence of the inhibitor were unable to develop embryogenic structures and its growth rate was diminished. This effect was reversible and concentration dependent. The results obtained contribute to the hypothesis that AOX activity supports metabolic reorganization as an essential part of cell reprogramming and, thus, enables restructuring and de novo cell differentiation.

  7. Activity and functional interaction of alternative oxidase and uncoupling protein in mitochondria from tomato fruit

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    F.E. Sluse


    Full Text Available Cyanide-resistant alternative oxidase (AOX is not limited to plant mitochondria and is widespread among several types of protists. The uncoupling protein (UCP is much more widespread than previously believed, not only in tissues of higher animals but also in plants and in an amoeboid protozoan. The redox energy-dissipating pathway (AOX and the proton electrochemical gradient energy-dissipating pathway (UCP lead to the same final effect, i.e., a decrease in ATP synthesis and an increase in heat production. Studies with green tomato fruit mitochondria show that both proteins are present simultaneously in the membrane. This raises the question of a specific physiological role for each energy-dissipating system and of a possible functional connection between them (shared regulation. Linoleic acid, an abundant free fatty acid in plants which activates UCP, strongly inhibits cyanide-resistant respiration mediated by AOX. Moreover, studies of the evolution of AOX and UCP protein expression and of their activities during post-harvest ripening of tomato fruit show that AOX and plant UCP work sequentially: AOX activity decreases in early post-growing stages and UCP activity is decreased in late ripening stages. Electron partitioning between the alternative oxidase and the cytochrome pathway as well as H+ gradient partitioning between ATP synthase and UCP can be evaluated by the ADP/O method. This method facilitates description of the kinetics of energy-dissipating pathways and of ATP synthase when state 3 respiration is decreased by limitation of oxidizable substrate.

  8. Alternative oxidase in the branched mitochondrial respiratory network: an overview on structure, function, regulation, and role

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    Sluse F.E.


    Full Text Available Plants and some other organisms including protists possess a complex branched respiratory network in their mitochondria. Some pathways of this network are not energy-conserving and allow sites of energy conservation to be bypassed, leading to a decrease of the energy yield in the cells. It is a challenge to understand the regulation of the partitioning of electrons between the various energy-dissipating and -conserving pathways. This review is focused on the oxidase side of the respiratory chain that presents a cyanide-resistant energy-dissipating alternative oxidase (AOX besides the cytochrome pathway. The known structural properties of AOX are described including transmembrane topology, dimerization, and active sites. Regulation of the alternative oxidase activity is presented in detail because of its complexity. The alternative oxidase activity is dependent on substrate availability: total ubiquinone concentration and its redox state in the membrane and O2 concentration in the cell. The alternative oxidase activity can be long-term regulated (gene expression or short-term (post-translational modification, allosteric activation regulated. Electron distribution (partitioning between the alternative and cytochrome pathways during steady-state respiration is a crucial measurement to quantitatively analyze the effects of the various levels of regulation of the alternative oxidase. Three approaches are described with their specific domain of application and limitations: kinetic approach, oxygen isotope differential discrimination, and ADP/O method (thermokinetic approach. Lastly, the role of the alternative oxidase in non-thermogenic tissues is discussed in relation to the energy metabolism balance of the cell (supply in reducing equivalents/demand in energy and carbon and with harmful reactive oxygen species formation.

  9. EPAct Alternative Fuel Transporation Program - State and Alternative Fuel Provider Fleets: Frequently Asked Questions

    Energy Technology Data Exchange (ETDEWEB)



    Factsheet answering frequently asked questions about the U.S. Department of Energy's Alternative Fuel Transportation Program (the Program) that implements provisions of Titles III–V of the Energy Policy Act of 1992 (EPAct). Answers to questions that are frequently asked about the Program by managers of state government and alternative fuel provider fleets are provided in the factsheet.

  10. The Chlamydomonas reinhardtii alternative oxidase 1 is regulated by heat stress. (United States)

    Zalutskaya, Zhanneta; Lapina, Tatiana; Ermilova, Elena


    The alternative oxidase (AOX) is a non-energy conserving terminal oxidase that has emerged as an important mitochondrial component of the cell stress responses. Although the most studied abiotic condition in relation to Chlamydomonas reinhardtii is high temperature, changes in AOX capacity of the alga were studied only under oxidative stress and cold. To examine whether elevated temperatures affected AOX1 expression, we applied quantitative real-time PCR and pharmaceutical approaches. In this work, we demonstrated a sharp increase in AOX1 transcript and protein abundance under heat stress. Furthermore, C. reinhardtii cells displayed a large increase in alternative respiration in response to high temperature. Feeding with the protein kinase inhibitor staurosporine strongly retarded the AOX1 transcription. Finally, the addition of the calcium chelator EGTA prevented heat-induced AOX1 expression. Together, our results imply that heat-inducible Ca(2+) influx and protein kinase(s) may mediate AOX1 expression at elevated temperatures. Characterization of heat-induced AOX1 regulation in the green alga C. reinhardtii provides a framework for a more complete understanding of the function of this conserved protein.

  11. Three Redox States of Trypanosoma brucei Alternative Oxidase Identified by Infrared Spectroscopy and Electrochemistry (United States)

    Maréchal, Amandine; Kido, Yasutoshi; Kita, Kiyoshi; Moore, Anthony L.; Rich, Peter R.


    Electrochemistry coupled with Fourier transform infrared (IR) spectroscopy was used to investigate the redox properties of recombinant alternative ubiquinol oxidase from Trypanosoma brucei, the organism responsible for African sleeping sickness. Stepwise reduction of the fully oxidized resting state of recombinant alternative ubiquinol oxidase revealed two distinct IR redox difference spectra. The first of these, signal 1, titrates in the reductive direction as an n = 2 Nernstian component with an apparent midpoint potential of 80 mV at pH 7.0. However, reoxidation of signal 1 in the same potential range under anaerobic conditions did not occur and only began with potentials in excess of 500 mV. Reoxidation by introduction of oxygen was also unsuccessful. Signal 1 contained clear features that can be assigned to protonation of at least one carboxylate group, further perturbations of carboxylic and histidine residues, bound ubiquinone, and a negative band at 1554 cm−1 that might arise from a radical in the fully oxidized protein. A second distinct IR redox difference spectrum, signal 2, appeared more slowly once signal 1 had been reduced. This component could be reoxidized with potentials above 100 mV. In addition, when both signals 1 and 2 were reduced, introduction of oxygen caused rapid oxidation of both components. These data are interpreted in terms of the possible active site structure and mechanism of oxygen reduction to water. PMID:19767647

  12. Questioning Needs Assessment: Some Limitations and Positive Alternatives (United States)

    Hannum, Wallace


    This article raises questions about the use of needs assessment by those seeking to improve the performance of organizations and individuals. While the discrepancy model of needs assessment has enjoyed a long history and is widely accepted, the author states that it has several undesirable consequences in terms of its impact. This article offers…

  13. Alternative Physics Examination Questions: Identification and Explanation of Different Discriminating Powers (United States)

    Forster, Patricia A.


    The issue of unfairness arises in high-stakes public examinations when students choose questions from alternatives that are offered and marks on the alternatives turn out to be discrepant. This paper addresses and defines unfairness and discrepancy in the context of alternative questions in Physics Tertiary Entrance Examinations (TEE) in Western Australia. As well, I present an analysis of question characteristics that explain observed marks-differences. The characteristics mainly relate to the construction of the text of questions, the detail on diagrams, and requirements for calculation. The list of characteristics could inform the setting of compulsory as well as alternative examination questions. The paper includes a brief exploration of results by gender on the alternative Physics TEE questions.

  14. Evidence of an alternative oxidase pathway for mitochondrial respiration in the scuticociliate Philasterides dicentrarchi. (United States)

    Mallo, Natalia; Lamas, Jesús; Leiro, José Manuel


    The presence of an alternative oxidase (AOX) in the mitochondria of the scuticociliate P. dicentrarchi was investigated. The mitochondrial oxygen consumption was measured in the presence of KCN, an inhibitor of cytochrome pathway (CP) respiration and salicylhydroxamic acid (SHAM), a specific inhibitor of alternative pathway (AP) respiration. AOX expression was monitored by western blotting with an AOX polyclonal antibody. The results showed that P. dicentrarchi possesses a branched mitochondrial electron transport chain with both cyanide-sensitive and -insensitive oxygen consumption. Mitochondrial respiration was partially inhibited by cyanide and completely inhibited by the combination of cyanide and SHAM, which is direct evidence for the existence of an AP in this ciliate. SHAM significantly inhibited in vitro growth of trophozoites both under normoxic and hypoxic conditions. AOX is a 42kD monomeric protein inducible by hypoxic conditions in experimental infections and by CP inhibitors such as cyanide and antimycin A, or by AP inhibitors such as SHAM. CP respiration was greatly stimulated during the exponential growth phase, while AP respiration increased during the stationary phase, in which AOX expression is induced. As the host does not possess AOX, and because during infection P. dicentrarchi respires via AP, it may be possible to develop inhibitors targeting the AP as a novel anti-scuticociliate therapy. Copyright © 2013 Elsevier GmbH. All rights reserved.

  15. The role of alternative oxidase in tomato fruit ripening and its regulatory interaction with ethylene. (United States)

    Xu, Fei; Yuan, Shu; Zhang, Da-Wei; Lv, Xin; Lin, Hong-Hui


    Although the alternative oxidase (AOX) has been proposed to play a role in fruit development, the function of AOX in fruit ripening is unclear. To gain further insight into the role of AOX in tomato fruit ripening, transgenic tomato plants 35S-AOX1a and 35S-AOX-RNAi were generated. Tomato plants with reduced LeAOX levels exhibited retarded ripening; reduced carotenoids, respiration, and ethylene production; and the down-regulation of ripening-associated genes. Moreover, no apparent respiratory climacteric occurred in the AOX-reduced tomato fruit, indicating that AOX might play an important role in climacteric respiration. In contrast, the fruit that overexpressed LeAOX1a accumulated more lycopene, though they displayed a similar pattern of ripening to wild-type fruit. Ethylene application promoted fruit ripening and anticipated ethylene production and respiration, including the alternative pathway respiration. Interestingly, the transgenic plants with reduced LeAOX levels failed to ripen after 1-methylcyclopropene (1-MCP) treatment, while such inhibition was notably less effective in 35S-AOX1a fruit. These findings indicate that AOX is involved in respiratory climacteric and ethylene-mediated fruit ripening of tomato.

  16. Fermentation and alternative oxidase contribute to the action of amino acid biosynthesis-inhibiting herbicides. (United States)

    Zulet, Amaia; Gil-Monreal, Miriam; Zabalza, Ana; van Dongen, Joost T; Royuela, Mercedes


    Acetolactate synthase inhibitors (ALS-inhibitors) and glyphosate (GLP) are two classes of herbicide that act by the specific inhibition of an enzyme in the biosynthetic pathway of branched-chain or aromatic amino acids, respectively. The physiological effects that are detected after application of these two classes of herbicides are not fully understood in relation to the primary biochemical target inhibition, although they have been well documented. Interestingly, the two herbicides' toxicity includes some common physiological effects suggesting that they kill the treated plants by a similar pattern despite targeting different enzymes. The induction of aerobic ethanol fermentation and alternative oxidase (AOX) are two examples of these common effects. The objective of this work was to gain further insight into the role of fermentation and AOX induction in the toxic consequences of ALS-inhibitors and GLP. For this, Arabidopsis T-DNA knockout mutants of alcohol dehydrogenase (ADH) 1 and AOX1a were used. The results found in wild-type indicate that both GLP and ALS-inhibitors reduce ATP production by inducing fermentation and alternative respiration. The main physiological effects in the process of herbicide activity upon treated plants were accumulation of carbohydrates and total free amino acids. The effects of the herbicides on these parameters were less pronounced in mutants compared to wild-type plants. The role of fermentation and AOX regarding pyruvate availability is also discussed.

  17. Expression of the alternative oxidase mitigates beta-amyloid production and toxicity in model systems. (United States)

    El-Khoury, Riyad; Kaulio, Eveliina; Lassila, Katariina A; Crowther, Damian C; Jacobs, Howard T; Rustin, Pierre


    Mitochondrial dysfunction has been widely associated with the pathology of Alzheimer's disease, but there is no consensus on whether it is a cause or consequence of disease, nor on the precise mechanism(s). We addressed these issues by testing the effects of expressing the alternative oxidase AOX from Ciona intestinalis, in different models of AD pathology. AOX can restore respiratory electron flow when the cytochrome segment of the mitochondrial respiratory chain is inhibited, supporting ATP synthesis, maintaining cellular redox homeostasis and mitigating excess superoxide production at respiratory complexes I and III. In human HEK293-derived cells, AOX expression decreased the production of beta-amyloid peptide resulting from antimycin inhibition of respiratory complex III. Because hydrogen peroxide was neither a direct product nor substrate of AOX, the ability of AOX to mimic antioxidants in this assay must be indirect. In addition, AOX expression was able to partially alleviate the short lifespan of Drosophila models neuronally expressing human beta-amyloid peptides, whilst abrogating the induction of markers of oxidative stress. Our findings support the idea of respiratory chain dysfunction and excess ROS production as both an early step and as a pathologically meaningful target in Alzheimer's disease pathogenesis, supporting the concept of a mitochondrial vicious cycle underlying the disease.

  18. Mitochondrial alternative oxidase maintains respiration and preserves photosynthetic capacity during moderate drought in Nicotiana tabacum. (United States)

    Dahal, Keshav; Wang, Jia; Martyn, Greg D; Rahimy, Farkhunda; Vanlerberghe, Greg C


    The mitochondrial electron transport chain includes an alternative oxidase (AOX) that is hypothesized to aid photosynthetic metabolism, perhaps by acting as an additional electron sink for photogenerated reductant or by dampening the generation of reactive oxygen species. Gas exchange, chlorophyll fluorescence, photosystem I (PSI) absorbance, and biochemical and protein analyses were used to compare respiration and photosynthesis of Nicotiana tabacum 'Petit Havana SR1' wild-type plants with that of transgenic AOX knockdown (RNA interference) and overexpression lines, under both well-watered and moderate drought-stressed conditions. During drought, AOX knockdown lines displayed a lower rate of respiration in the light than the wild type, as confirmed by two independent methods. Furthermore, CO2 and light response curves indicated a nonstomatal limitation of photosynthesis in the knockdowns during drought, relative to the wild type. Also relative to the wild type, the knockdowns under drought maintained PSI and PSII in a more reduced redox state, showed greater regulated nonphotochemical energy quenching by PSII, and displayed a higher relative rate of cyclic electron transport around PSI. The origin of these differences may lie in the chloroplast ATP synthase amount, which declined dramatically in the knockdowns in response to drought. None of these effects were seen in plants overexpressing AOX. The results show that AOX is necessary to maintain mitochondrial respiration during moderate drought. In its absence, respiration rate slows and the lack of this electron sink feeds back on the photosynthetic apparatus, resulting in a loss of chloroplast ATP synthase that then limits photosynthetic capacity.

  19. Regulation of Thermogenesis In Plants: The Interaction of Alternative Oxidase and Plant Uncoupling Mitochondrial Protein

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    Yan Zhu; Jianfei Lu; Jing Wang; Fu Chen; Feifan Leng; Hongyu Li


    Thermogenesis is a process of heat production in living organisms.It is rare in plants,but it does occur in some species of angiosperm.The heat iS generated via plant mitochondrial respiration.As possible Involvement in thermogenesis of mitochondrial factors,alternative oxidases(AOXs)and plant uncoupling mitochondrial proteins(PUMPs)have been well studied.AOXs and PUMPs are ubiquitously present in the inner membrane of plant mitochondria.They serve as two major energy dissipation systems that balance mitochondrial respiration and uncoupled phosphorylation by dissipating the H+ redox energy and proton electrochemical gradient(△μH+)as heat,respectively.AOXs and PUMPs exert similar physiological functions during homeothermic heat production in thermogenic plants.AOXs have five isoforms,while PUMPs have six.Both AOXS and PUMPS are encoded by small nuclear multigene families.Multiple isoforms are expressed in different tissues or organs.Extensive studies have been done in the area of thermogenesis in higher plants.In this review,we focus on the involvement and regulation of AOXs and PUMPs in thermogenesis.

  20. Urate oxidase purification by salting-in crystallization: towards an alternative to chromatography.

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    Marion Giffard

    Full Text Available BACKGROUND: Rasburicase (Fasturtec® or Elitek®, Sanofi-Aventis, the recombinant form of urate oxidase from Aspergillus flavus, is a therapeutic enzyme used to prevent or decrease the high levels of uric acid in blood that can occur as a result of chemotherapy. It is produced by Sanofi-Aventis and currently purified via several standard steps of chromatography. This work explores the feasibility of replacing one or more chromatography steps in the downstream process by a crystallization step. It compares the efficacy of two crystallization techniques that have proven successful on pure urate oxidase, testing them on impure urate oxidase solutions. METHODOLOGY/PRINCIPAL FINDINGS: Here we investigate the possibility of purifying urate oxidase directly by crystallization from the fermentation broth. Based on attractive interaction potentials which are known to drive urate oxidase crystallization, two crystallization routes are compared: a by increased polymer concentration, which induces a depletion attraction and b by decreased salt concentration, which induces attractive interactions via a salting-in effect. We observe that adding polymer, a very efficient way to crystallize pure urate oxidase through the depletion effect, is not an efficient way to grow crystals from impure solution. On the other hand, we show that dialysis, which decreases salt concentration through its strong salting-in effect, makes purification of urate oxidase from the fermentation broth possible. CONCLUSIONS: The aim of this study is to compare purification efficacy of two crystallization methods. Our findings show that crystallization of urate oxidase from the fermentation broth provides purity comparable to what can be achieved with one chromatography step. This suggests that, in the case of urate oxidase, crystallization could be implemented not only for polishing or concentration during the last steps of purification, but also as an initial capture step, with minimal

  1. Routing of Hansenula polymorpha alcohol oxidase : An alternative peroxisomal protein-sorting machinery

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    Gunkel, Katja; Dijk, Ralf van; Veenhuis, Marten; Klei, Ida J. van der

    Import of Hansenula polymorpha alcohol oxidase (AO) into peroxisomes is dependent on the PTS1 receptor, HpPex5p. The PTS1 of AO (-LARF) is sufficient to direct reporter proteins to peroxisomes. To study AO sorting in more detail, strains producing mutant AO proteins were constructed. AO containing a

  2. Effects of Enhanced UV-B Radiation on the Activity and Expression of Alternative Oxidase in Red Kidney Bean Leaves

    Institute of Scientific and Technical Information of China (English)

    Ming-Guang Zhao; Ying-Gao Liu; Li-Xin Zhang; Lin Zheng; Yu-Rong Bi


    An increase in ultraviolet (UV) B radiation on the earth's surface is a feature of current global climate changes. It has been reported that alternative oxidase (AOX) may have a protective role against oxidative stress induced by environmental stresses, such as UV-B. To better understand the characteristic tolerance of plants to UV-B radiation, the effects of enhanced UV-B radiation on the activity and expression of AOX in red kidney bean (Phaseolus vulgaris) leaves were investigated in the present study. The results show that the total respiration rate and AOX activity in red kidney bean leaves increased significantly during treatment with enhanced UV-B. However, cytochrome oxidase (COX) activity did not change at 24 h of UV-B treatment, before dropping rapidly. Both alternative pathway content and alternative pathway activity were increased in the presence of exogenous H2O2. Immunoblotting analysis with anti-AOX monoclonal antibody revealed that expression of the AOX protein increased in red kidney bean leaves under enhanced UV-B radiation, reaching a peak at 72increase in AOX activity in red kidney bean leaves under enhanced UV-B radiation was mainly due to H2O2-induced AOX expression.

  3. Importance of the alternative oxidase (AOX) pathway in regulating cellular redox and ROS homeostasis to optimize photosynthesis during restriction of the cytochrome oxidase pathway in Arabidopsis thaliana. (United States)

    Vishwakarma, Abhaypratap; Tetali, Sarada Devi; Selinski, Jennifer; Scheibe, Renate; Padmasree, Kollipara


    The importance of the alternative oxidase (AOX) pathway, particularly AOX1A, in optimizing photosynthesis during de-etiolation, under elevated CO2, low temperature, high light or combined light and drought stress is well documented. In the present study, the role of AOX1A in optimizing photosynthesis was investigated when electron transport through the cytochrome c oxidase (COX) pathway was restricted at complex III. Leaf discs of wild-type (WT) and aox1a knock-out mutants of Arabidopsis thaliana were treated with antimycin A (AA) under growth-light conditions. To identify the impact of AOX1A deficiency in optimizing photosynthesis, respiratory O2 uptake and photosynthesis-related parameters were measured along with changes in redox couples, reactive oxygen species (ROS), lipid peroxidation and expression levels of genes related to respiration, the malate valve and the antioxidative system. In the absence of AA, aox1a knock-out mutants did not show any difference in physiological, biochemical or molecular parameters compared with WT. However, after AA treatment, aox1a plants showed a significant reduction in both respiratory O2 uptake and NaHCO3-dependent O2 evolution. Chlorophyll fluorescence and P700 studies revealed that in contrast to WT, aox1a knock-out plants were incapable of maintaining electron flow in the chloroplastic electron transport chain, and thereby inefficient heat dissipation (low non-photochemical quenching) was observed. Furthermore, aox1a mutants exhibited significant disturbances in cellular redox couples of NAD(P)H and ascorbate (Asc) and consequently accumulation of ROS and malondialdehyde (MDA) content. By contrast, WT plants showed a significant increase in transcript levels of CSD1, CAT1, sAPX, COX15 and AOX1A in contrast to aox1a mutants. These results suggest that AOX1A plays a significant role in sustaining the chloroplastic redox state and energization to optimize photosynthesis by regulating cellular redox homeostasis and ROS

  4. Alternative Splicing and Differential Expression of Two Transcripts of Nicotine Adenine Dinucleotide Phosphate Oxidase B Gene from Zea mays

    Institute of Scientific and Technical Information of China (English)

    Fan Lin; Yun Zhang; Ming-Yi Jiang


    With the exception of rice, little is known about the existence of respiratory burst oxidase homolog (rboh) gene in cereals. The present study reports the cloning and analysis of a novel rboh gene, termed ZmrbohB, from maize (Zea mays L.). The full-length cDNA of ZmrbohB encodes a 942 amino acid protein containing all of the respiratory burst oxidase homolog catalytically critical motifs.Altemative splicing of ZmrbohB has generated two transcript isoforms, ZmrbohB-α and -β. Spliced transcript ZmrbohB-β retains an unspliced intron 11 that carries a premature termination codon and probably leads to nonsense-mediated mRNA decay. Expression analysis showed that two splice isoforms were differentially expressed in various tissues and at different developmental stages, and the major product was ZmrbohB-α. The transcripts of ZmrbohB-α accumulated markedly when the maize seedlings were subjected to various abiotic stimuli, such as wounding, cold (4℃), heat (40℃), UV and salinity stress. In addition, several abiotic stimuli also affected the alternative splicing pattern of ZmrbohB except wounding. These results provide new insight into roles in the expression regulation of plant rboh genes and suggest that ZmrbohB gene may play a role in response to environmental stresses.

  5. Alternative Oxidase: A Mitochondrial Respiratory Pathway to Maintain Metabolic and Signaling Homeostasis during Abiotic and Biotic Stress in Plants

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    Greg C. Vanlerberghe


    Full Text Available Alternative oxidase (AOX is a non-energy conserving terminal oxidase in the plant mitochondrial electron transport chain. While respiratory carbon oxidation pathways, electron transport, and ATP turnover are tightly coupled processes, AOX provides a means to relax this coupling, thus providing a degree of metabolic homeostasis to carbon and energy metabolism. Beside their role in primary metabolism, plant mitochondria also act as “signaling organelles”, able to influence processes such as nuclear gene expression. AOX activity can control the level of potential mitochondrial signaling molecules such as superoxide, nitric oxide and important redox couples. In this way, AOX also provides a degree of signaling homeostasis to the organelle. Evidence suggests that AOX function in metabolic and signaling homeostasis is particularly important during stress. These include abiotic stresses such as low temperature, drought, and nutrient deficiency, as well as biotic stresses such as bacterial infection. This review provides an introduction to the genetic and biochemical control of AOX respiration, as well as providing generalized examples of how AOX activity can provide metabolic and signaling homeostasis. This review also examines abiotic and biotic stresses in which AOX respiration has been critically evaluated, and considers the overall role of AOX in growth and stress tolerance.

  6. Enhanced fatty acid accumulation in Isochrysis galbana by inhibition of the mitochondrial alternative oxidase pathway under nitrogen deprivation. (United States)

    Zhang, Litao; Liu, Jianguo


    The purpose of this study was to clarify the interrelation between the mitochondrial alternative oxidase (AOX) pathway and fatty acid accumulation in marine microalga Isochrysis galbana. Under normal conditions, the activity of the AOX pathway was maintained at a low level in I. galbana. Compared with the normal condition, nitrogen deprivation significantly increased the AOX pathway activity and fatty acid accumulation. Under nitrogen deprivation, the inhibition of the AOX pathway by salicylhydroxamic acid caused the accumulation of reducing equivalents and the over-reduction of chloroplasts in I. galbana cells, leading to a decrease in the photosynthetic O2 evolution rate. The over-production of reducing equivalents due to the inhibition of the AOX pathway under nitrogen deprivation further enhanced the accumulation of fatty acids in I. galbana cells.

  7. High Osmolarity Environments Activate the Mitochondrial Alternative Oxidase in Debaryomyces Hansenii (United States)

    Garcia-Neto, Wilson; Cabrera-Orefice, Alfredo; Uribe-Carvajal, Salvador; Kowaltowski, Alicia J.; Alberto Luévano-Martínez, Luis


    The oleaginous yeast Debaryomyces hansenii is a good model to understand molecular mechanisms involved in halotolerance because of its impressive ability to survive under a wide range of salt concentrations. Several cellular adaptations are implicated in this response, including the presence of a cyanide-insensitive ubiquinol oxidase (Aox). This protein, which is present in several taxonomical orders, has been related to different stress responses. However, little is known about its role in mitochondria during transitions from low to high saline environments. In this report, we analyze the effects of Aox in shifts from low to high salt concentrations in the culture media. At early stages of a salt insult, we observed that this protein prevents the overflow of electrons on the mitochondrial respiratory chain, thus, decreasing the production of reactive oxygen species. Interestingly, in the presence of high osmolite concentrations, Aox activity is able to sustain a stable membrane potential when coupled to complex I, despite a compromised cytochrome pathway. Taken together, our results suggest that under high osmolarity conditions Aox plays a critical role regulating mitochondrial physiology. PMID:28060946

  8. Characterization of alternative molecular forms of xanthine oxidase in the mouse. (United States)

    Duke, E J; Joyce, P; Ryan, J P


    1. Two major forms of xanthine oxidase are demonstrated for the mouse. On polyacrylamide-gel electrophoresis the duodenal form migrates faster towards the anode than that of the liver. Both forms also differ in their (NH(4))(2)SO(4) precipitation patterns and sucrose-density-gradient molecular-weight determinations. 2. The liver form is fully converted into the duodenal form by incubation at 37 degrees C with 2.5mg of crude trypsin/ml for 1(1/2)h, without loss of activity. The trypsin-treated liver form behaves like the normal duodenal form as characterized by electrophoresis, (NH(4))(2)SO(4) precipitation patterns, and sucrose-density-gradient molecular-weight determinations. 3. Partial conversion is also brought about by purified trypsin and chymotrypsin, but not with beta-carboxypeptidase or lipase. The conversion is inhibited by soya-bean trypsin inhibitor. 4. In embryo mice the duodenal form is similar to the liver form on electrophoresis. 5. These studies indicate, as might be expected, that the duodenal form is a modified version of the liver enzyme, probably caused by proteolytic alteration.

  9. Asking well-built questions for evidence-based practice in augmentative and alternative communication. (United States)

    Schlosser, Ralf W; Koul, Rajinder; Costello, John


    Evidence-based practice (EBP) is increasingly being advocated as the preferred approach to practice in augmentative and alternative communication (AAC). The EBP process involves multiple steps. The asking of a well-built question is the first step in the quest for answers. At the same time it is also often the first stumbling block for practitioners. To facilitate the asking of well-built questions it may be helpful to follow a template. The most frequently used template is PICO, which stands for patient, intervention, comparison, and outcome [Richardson, W., Wilson, M., Nishikawa, J., & Hayward, R. (1995). The well-built clinical question: A key to evidence-based decisions. ACP Journal Club, 123, A12-A13]. In this article, we examine the suitability of the PICO template for AAC in terms of the representativeness of the components, and the appropriateness of its subcomponents, and their terminology. Based on this analysis, we propose the PESICO template, which stands for person, environments, stakeholders, intervention, comparison, and outcome. This template is then illustrated with examples representing a range of decision-making areas in AAC. Finally, directions for future research are provided. The reader will be able to: (1) appreciate the importance of asking well-built questions, (2) name the shortcomings of the PICO template, and (3) describe the components of the proposed PESICO template for asking well-built questions.

  10. Bilirubin Oxidase from Myrothecium verrucaria Physically Absorbed on Graphite Electrodes. Insights into the Alternative Resting Form and the Sources of Activity Loss (United States)

    Tasca, Federico; Farias, Diego; Castro, Carmen; Acuna-Rougier, Cristina; Antiochia, Riccarda


    The oxygen reduction reaction is one of the most important chemical processes in energy converting systems and living organisms. Mediator-less, direct electro-catalytic reduction of oxygen to water was achieved on spectrographite electrodes modified by physical adsorption of bilirubin oxidases from Myrothecium verrucaria. The existence of an alternative resting form of the enzyme is validated. The effect on the catalytic cycle of temperature, pH and the presence of halogens in the buffer was investigated. Previous results on the electrochemistry of bilirubin oxidase and on the impact of the presence of halogens are reviewed and reinterpreted. PMID:26196288

  11. Changes of alternative oxidase activity, capacity and protein content in leaves of Cucumis sativus wild-type and MSC16 mutant grown under different light intensities. (United States)

    Florez-Sarasa, Igor; Ostaszewska, Monika; Galle, Alexander; Flexas, Jaume; Rychter, Anna M; Ribas-Carbo, Miquel


    In vitro studies demonstrated that alternative oxidase (AOX) is biochemically regulated by a sulfhydryl-disulfide system, interaction with alpha-ketoacids, ubiquinone pool redox state and protein content among others. However, there is still scarce information about the in vivo regulation of the AOX. Cucumis sativus wild-type (WT) and MSC16 mutant plants were grown under two different light intensities and were used to analyze the relationship between the amount of leaf AOX protein and its in vivo capacity and activity at night and day periods. WT and MSC16 plants presented lower total respiration (V(t)), cytochrome oxidase pathway (COP) activity (v(cyt)) and alternative oxidase pathway (AOP) activity (v(alt)) when grown at low light (LL), although growth light intensity did not change the amount of cytochrome oxidase (COX) nor AOX protein. Changes of v(cyt) related to growing light conditions suggested a substrate availability and energy demand control. On the other hand, the total amount of AOX protein present in the tissue does not play a role in the regulation neither of the capacity nor of the activity of the AOP in vivo. Soluble carbohydrates were directly related to the activity of the AOP. However, although differences in soluble sugar contents mostly regulate the capacity of the AOP at different growth light intensities, additional regulatory mechanisms are necessary to fully explain the observed results.

  12. Light Intensity Affects Chlorophyll Synthesis During Greening Process by Metabolite Signal from Mitochondrial Alternative Oxidase in Arabidopsis

    Institute of Scientific and Technical Information of China (English)

    ZHANG; Dawei; YUAN; Shu; 徐飞; ZHU; Feng; YUAN; Ming; YE; Huaxun; GUO; Hongqing; LV; Xin; YIN; Yanhai; 林宏辉


    Although mitochondrial alternative oxidase(AOX)has been proposed to play essential roles in high light stress tolerance,the effects of AOX on chlorophyll synthesis are unclear.Previous studies indicated that during greening,chlorophyll accumulation was largely delayed in plants whose mitochondrial cyanide-resistant respiration was inhibited by knocking out nuclear encoded AOX gene.Here we show that this delay of chlorophyll accumulation was more significant under high light condition.Inhibition of cyanide-resistant respiration was also accompanied by the increase of plastid NADPH/NADP~+ratio,especially under high light treatment which subsequently blocked the import of multiple plastidial proteins,such as some components of the photosynthetic electron transport chain,the Calvin-Benson cycle enzymes and malate/oxaloacetate shuttle components.Over expression of AOXla rescued the aoxla mutant phenotype,including the chlorophyll accumulation during greening and plastidial protein import.It thus suggests that light intensity affects chlorophyll synthesis during greening process by a metabolic signal,the AOX-derived plastidial NADPH/NADP~+ratio change.And our results thus revealed a molecular mechanism of chloroplast-mitochondria interactions.

  13. Alternative Oxidase Activity in Tobacco Leaf Mitochondria (Dependence on Tricarboxylic Acid Cycle-Mediated Redox Regulation and Pyruvate Activation). (United States)

    Vanlerberghe, G. C.; Day, D. A.; Wiskich, J. T.; Vanlerberghe, A. E.; McIntosh, L.


    Transgenic Nicotiana tabacum (cv Petit Havana SR1) containing high levels of mitochondrial alternative oxidase (AOX) protein due to the introduction of a sense transgene(s) of Aox1, the nuclear gene encoding AOX, were used to investigate mechanisms regulating AOX activity. After purification of leaf mitochondria, a large proportion of the AOX protein was present as the oxidized (covalently associated and less active) dimer. High AOX activity in these mitochondria was dependent on both reduction of the protein by DTT (to the noncovalently associated and more active dimer) and its subsequent activation by certain [alpha]-keto acids, particularly pyruvate. Reduction of AOX to its more active form could also be mediated by intramitochondrial reducing power generated by the oxidation of certain tricarboxylic acid cycle substrates, most notably isocitrate and malate. Our evidence suggests that NADPH may be specifically required for AOX reduction. All of the above regulatory mechanisms applied to AOX in wild-type mitochondria as well. Transgenic leaves lacking AOX due to the introduction of an Aox1 antisense transgene or multiple sense transgenes were used to investigate the potential physiological significance of the AOX-regulatory mechanisms. Under conditions in which respiratory carbon metabolism is restricted by the capacity of mitochondrial electron transport, feed-forward activation of AOX by mitochondrial reducing power and pyruvate may act to prevent redirection of carbon metabolism, such as to fermentative pathways.

  14. The role of alternative oxidase in modulating carbon use efficiency and growth during macronutrient stress in tobacco cells. (United States)

    Sieger, Stephen M; Kristensen, Brian K; Robson, Christine A; Amirsadeghi, Sasan; Eng, Edward W Y; Abdel-Mesih, Amal; Møller, Ian M; Vanlerberghe, Greg C


    When wild-type (wt) tobacco (Nicotiana tabacum cv. Petit Havana SR1) cells are grown under macronutrient (P or N) limitation, they induce large amounts of alternative oxidase (AOX), which constitutes a non-energy-conserving branch of the respiratory electron transport chain. To investigate the significance of AOX induction, wt cells were compared with transgenic (AS8) cells lacking AOX. Under nutrient limitation, growth of wt cell cultures was dramatically reduced and carbon use efficiency (g cell dry weight gain g(-1) sugar consumed) decreased by 42-63%. However, the growth of AS8 was only moderately reduced by the nutrient deficiencies and carbon use efficiency values remained the same as under nutrient-sufficient conditions. As a result, the nutrient limitations more severely compromised the tissue nutrient status (P or N) of AS8 than wt cells. Northern analyses and a comparison of the mitochondrial protein profiles of wt and AS8 cells indicated that the lack of AOX in AS8 under P limitation was associated with increased levels of proteins commonly associated with oxidative stress and/or stress injury. Also, the level of electron transport chain components was consistently reduced in AS8 while tricarboxylic acid cycle enzymes did not show a universal trend in abundance in comparison to the wt. Alternatively, the lack of AOX in AS8 cells under N limitation resulted in enhanced carbohydrate accumulation. It is concluded that AOX respiration provides an important general mechanism by which plant cells can modulate their growth in response to nutrient availability and that AOX also has nutrient-specific roles in maintaining cellular redox and carbon balance.

  15. A classification scheme for alternative oxidases reveals the taxonomic distribution and evolutionary history of the enzyme in angiosperms. (United States)

    Costa, José Hélio; McDonald, Allison E; Arnholdt-Schmitt, Birgit; Fernandes de Melo, Dirce


    A classification scheme based on protein phylogenies and sequence harmony method was used to clarify the taxonomic distribution and evolutionary history of the alternative oxidase (AOX) in angiosperms. A large data set analyses showed that AOX1 and AOX2 subfamilies were distributed into 4 phylogenetic clades: AOX1a-c/1e, AOX1d, AOX2a-c and AOX2d. High diversity in AOX family compositions was found. While the AOX2 subfamily was not detected in monocots, the AOX1 subfamily has expanded (AOX1a-e) in the large majority of these plants. In addition, Poales AOX1b and 1d were orthologous to eudicots AOX1d and then renamed as AOX1d1 and 1d2. AOX1 or AOX2 losses were detected in some eudicot plants. Several AOX2 duplications (AOX2a-c) were identified in eudicot species, mainly in the asterids. The AOX2b originally identified in eudicots in the Fabales order (soybean, cowpea) was divergent from AOX2a-c showing some specific amino acids with AOX1d and then it was renamed as AOX2d. AOX1d and AOX2d seem to be stress-responsive, facultative and mutually exclusive among species suggesting a complementary role with an AOX1(a) in stress conditions. Based on the data collected, we present a model for the evolutionary history of AOX in angiosperms and highlight specific areas where further research would be most beneficial. Copyright © 2014 Elsevier B.V. All rights reserved.

  16. Modified expression of alternative oxidase in transgenic tomato and petunia affects the level of tomato spotted wilt virus resistance

    Directory of Open Access Journals (Sweden)

    Ma Hao


    Full Text Available Abstract Background Tomato spotted wilt virus (TSWV has a very wide host range, and is transmitted in a persistent manner by several species of thrips. These characteristics make this virus difficult to control. We show here that the over-expression of the mitochondrial alternative oxidase (AOX in tomato and petunia is related to TSWV resistance. Results The open reading frame and full-length sequence of the tomato AOX gene LeAox1au were cloned and introduced into tomato 'Healani' and petunia 'Sheer Madness' using Agrobacterium-mediated transformation. Highly expressed AOX transgenic tomato and petunia plants were selfed and transgenic R1 seedlings from 10 tomato lines and 12 petunia lines were used for bioassay. For each assayed line, 22 to 32 tomato R1 progeny in three replications and 39 to 128 petunia progeny in 13 replications were challenged with TSWV. Enzyme-Linked Immunosorbent Assays showed that the TSWV levels in transgenic tomato line FKT4-1 was significantly lower than that of wild-type controls after challenge with TSWV. In addition, transgenic petunia line FKP10 showed significantly less lesion number and smaller lesion size than non-transgenic controls after inoculation by TSWV. Conclusion In all assayed transgenic tomato lines, a higher percentage of transgenic progeny had lower TSWV levels than non-transgenic plants after challenge with TSWV, and the significantly increased resistant levels of tomato and petunia lines identified in this study indicate that altered expression levels of AOX in tomato and petunia can affect the levels of TSWV resistance.

  17. Studies on the relationship between cyanide-resistant respi-ration and expression of alternative oxidase in mung bean using antibodies prepared by synthetic polypeptide

    Institute of Scientific and Technical Information of China (English)

    LI; Chijun; (


    [1]Liang, Z., Liang, H. G., The respiratory metabolism of plants, in Plant Physiology and Molecular Biology (eds. Yu, S. W., Tang, Z. C.) (in Chinese), 2nd ed., Beijing: Science Press, 1998, 344-365.[2]Lü, C. S., Liang, H. G., Induced respiration in melon fruits, Scientia Sinica, 1963, 12(4): 616.[3]Liang, H. G., Lü, C. S., A comparative study of CN-resistant respiration in different cultures of tobacco callus, Plant Physiol., 1984, 75: 876.[4]Elthon, T. E., McIntosh, L., Identification of the alternative terminal oxidase of higher plant mitochondria, Proc. Natl. Acad. Sci. USA, 1987, 84: 8399.[5]Elthon, T. E., Nickels, R. L., McIntosh, L., Monoclonal antibodies to the alternative oxidase of higher plant mitochondria, Plant Physiol., 1989, 89: 1311.[6]Liang, W. S., Liang, H. G., Progress of the alternative oxidase, Chinese Bulletin of Botany (in Chinese), 1997, 14(2): 9.[7]Liang, W. S., Liang, H. G., Induction of alternative oxidase expression by endogenous ethylene in aging potato slices, Acta Phytophysiol. Sin. (in Chinese), 1999, 25(2): 205.[8]He, J. X., Wei, Z. Q., Liang, H. G., Effects of water stress on development, and operation and gene expression of cyanide-resistant respiratory pathway in wheat, Science in China, Ser. C, 1999, 42(3): 300.[9]McIntosh, L., Molecular biology of the alternative oxidase, Plant Physiol., 1994, 105: 781.[10] Wang, J., Zhang, L. X., Liu, Z. L. et al., A possible calcium binding site in D1 protein: A fluorescence and FTIR study of the interaction between lanthanides and a synthetic peptide, Photosynthesis Research, 1995, 44: 297.[11] Li, X. P., Du, L. F., Liang, H. G. et al., Preparation and identification of antidodecapeptide of polypeptide D1 or photosys-tem II reaction center, Prog. Biochem. Biophys. (in Chinese), 1997, 24(3): 283.[12] Wen, J. Q., Liang, H. G., Studies on energy status and mitochondria respiration during growth and senescence of mung bean cotyledons, Physiol

  18. Eye tracking research to answer questions about augmentative and alternative communication assessment and intervention. (United States)

    Wilkinson, Krista M; Mitchell, Teresa


    Recently, eye tracking technologies (i.e., technologies that automatically track the point of an individual's gaze while that person views or interacts with a visual image) have become available for research purposes. Based on the sampling of the orientation of the individual's eyes, researchers can quantify which locations within the visual image were fixated (viewed), for how long, and how many times. These automated eye tracking research technologies open up a wealth of avenues for investigating how individuals with developmental or acquired communication disabilities may respond to aided augmentative and alternative communication (AAC) systems. In this paper, we introduce basic terminology and explore some of the special challenges of conducting eye tracking research with populations with disabilities who might use AAC, including challenges of inferring attention from the presence of fixation and challenges related to calibration that may result from participant characteristics, behavioral idiosyncracies, and/or the number of calibration points. We also examine how the technology can be applied to ask well-structured experimental questions that have direct clinical relevance, with a focus on the unique contributions that eye tracking research can provide by (a) allowing evaluation of skills in individuals who are difficult to assess via traditional methods, and (b) facilitating access to information on underlying visual cognitive processes that is not accessible via traditional behavioral measures.

  19. Functional expression of the Acanthamoeba castellanii alternative oxidase in Escherichia coli; regulation of the activity and evidence for Acaox gene function. (United States)

    Antos-Krzeminska, Nina; Jarmuszkiewicz, Wieslawa


    To evidence Acanthamoeba castellanii alternative oxidase (AcAOX) gene product function, we studied alterations in the levels of mRNA and protein and AcAOX activity during growth in amoeba batch culture. Moreover, heterologous expression of AcAOX in AOX-deficient Escherichia coli confirmed by the protein immunodetection and functional studies was performed. Despite the presence of native bo and bd quinol oxidases in E. coli membrane, from which the latter is known to be cyanide-resistant, functional expression of AcAOX in E. coli conferred cyanide-resistant benzohydroxamate-sensitive respiration on the bacteria. Moreover, AcAOX activity in transformed bacteria was stimulated by GMP and inhibited by ATP, indicating that AcAOX is regulated by mutual exclusion of purine nucleotides, which was previously demonstrated in the mitochondria of A. castellanii.

  20. Alternative Oxidase Gene Family in Hypericum perforatum L.: Characterization and Expression at the Post-germinative Phase (United States)

    Velada, Isabel; Cardoso, Hélia G.; Ragonezi, Carla; Nogales, Amaia; Ferreira, Alexandre; Valadas, Vera; Arnholdt-Schmitt, Birgit


    Alternative oxidase (AOX) protein is located in the inner mitochondrial membrane and is encoded in the nuclear genome being involved in plant response upon a diversity of environmental stresses and also in normal plant growth and development. Here we report the characterization of the AOX gene family of Hypericum perforatum L. Two AOX genes were identified, both with a structure of four exons (HpAOX1, acc. KU674355 and HpAOX2, acc. KU674356). High variability was found at the N-terminal region of the protein coincident with the high variability identified at the mitochondrial transit peptide. In silico analysis of regulatory elements located at intronic regions identified putative sequences coding for miRNA precursors and trace elements of a transposon. Simple sequence repeats were also identified. Additionally, the mRNA levels for the HpAOX1 and HpAOX2, along with the ones for the HpGAPA (glyceraldehyde-3-phosphate dehydrogenase A subunit) and the HpCAT1 (catalase 1), were evaluated during the post-germinative development. Gene expression analysis was performed by RT-qPCR with accurate data normalization, pointing out HpHYP1 (chamba phenolic oxidative coupling protein 1) and HpH2A (histone 2A) as the most suitable reference genes (RGs) according to GeNorm algorithm. The HpAOX2 transcript demonstrated larger stability during the process with a slight down-regulation in its expression. Contrarily, HpAOX1 and HpGAPA (the corresponding protein is homolog to the chloroplast isoform involved in the photosynthetic carbon assimilation in other plant species) transcripts showed a marked increase, with a similar expression pattern between them, during the post-germinative development. On the other hand, the HpCAT1 (the corresponding protein is homolog to the major H2O2-scavenging enzyme in other plant species) transcripts showed an opposite behavior with a down-regulation during the process. In summary, our findings, although preliminary, highlight the importance to

  1. Alternative Oxidase Gene Family in Hypericum perforatum L.: Characterization and Expression at the Post-germinative Phase. (United States)

    Velada, Isabel; Cardoso, Hélia G; Ragonezi, Carla; Nogales, Amaia; Ferreira, Alexandre; Valadas, Vera; Arnholdt-Schmitt, Birgit


    Alternative oxidase (AOX) protein is located in the inner mitochondrial membrane and is encoded in the nuclear genome being involved in plant response upon a diversity of environmental stresses and also in normal plant growth and development. Here we report the characterization of the AOX gene family of Hypericum perforatum L. Two AOX genes were identified, both with a structure of four exons (HpAOX1, acc. KU674355 and HpAOX2, acc. KU674356). High variability was found at the N-terminal region of the protein coincident with the high variability identified at the mitochondrial transit peptide. In silico analysis of regulatory elements located at intronic regions identified putative sequences coding for miRNA precursors and trace elements of a transposon. Simple sequence repeats were also identified. Additionally, the mRNA levels for the HpAOX1 and HpAOX2, along with the ones for the HpGAPA (glyceraldehyde-3-phosphate dehydrogenase A subunit) and the HpCAT1 (catalase 1), were evaluated during the post-germinative development. Gene expression analysis was performed by RT-qPCR with accurate data normalization, pointing out HpHYP1 (chamba phenolic oxidative coupling protein 1) and HpH2A (histone 2A) as the most suitable reference genes (RGs) according to GeNorm algorithm. The HpAOX2 transcript demonstrated larger stability during the process with a slight down-regulation in its expression. Contrarily, HpAOX1 and HpGAPA (the corresponding protein is homolog to the chloroplast isoform involved in the photosynthetic carbon assimilation in other plant species) transcripts showed a marked increase, with a similar expression pattern between them, during the post-germinative development. On the other hand, the HpCAT1 (the corresponding protein is homolog to the major H2O2-scavenging enzyme in other plant species) transcripts showed an opposite behavior with a down-regulation during the process. In summary, our findings, although preliminary, highlight the importance to

  2. Alternative oxidase gene family in Hypericum perforatum L.: characterization and expression at the post-germinative phase

    Directory of Open Access Journals (Sweden)

    Isabel Velada


    Full Text Available Alternative oxidase (AOX protein is located in the inner mitochondrial membrane and is encoded in the nuclear genome being involved in plant response upon diversity of environmental stresses and also in normal plant growth and development. Here we report the characterization of the AOX gene family of Hypericum perforatum L. Two AOX genes were identified, both with a structure of four exons (HpAOX1, acc. KU674355 and HpAOX2, acc. KU674356. High variability was found at the N-terminal region of the protein coincident with the high variability identified at the mitochondrial transit peptide. In silico analysis of regulatory elements located at intronic regions identified putative sequences coding for miRNA precursors and trace elements of a transposon. Simple sequence repeats were also identified. Additionally, the mRNA levels for the HpAOX1 and HpAOX2, along with the ones for the HpGAPA and the HpCAT1, were evaluated during the post-germinative development. The gene expression analysis was performed by RT-qPCR with accurate data normalization, pointing out HpHYP1 and HpH2A as the most suitable RGs according to GeNorm algorithm. The HpAOX2 transcript demonstrated larger stability during the process with a slight down-regulation in its expression. Contrarily, HpAOX1 and HpGAPA (the corresponding protein is homolog to the chloroplast isoform involved in the photosynthetic carbon assimilation in other species transcripts showed a marked increase, with a similar expression pattern between them, during the post-germinative development. On the other hand, the HpCAT1 (the corresponding protein is homolog to the major H2O2-scavenging enzyme in other species transcripts showed an opposite behavior with a down-regulation during the process. In summary, our findings, although preliminary, highlight the importance to investigate in more detail the participation of AOX genes during the post-germinative development in Hypericum, in order to explore their

  3. Alternative oxidase pathway optimizes photosynthesis during osmotic and temperature stress by regulating cellular ROS, malate valve and antioxidative systems

    Directory of Open Access Journals (Sweden)



    Full Text Available The present study reveals the importance of alternative oxidase (AOX pathway in optimizing photosynthesis under osmotic and temperature stress conditions in the mesophyll protoplasts of Pisum sativum. The responses of photosynthesis and respiration were monitored at saturating light intensity of 1000 µmoles m-2 s-1 at 25 oC under a range of sorbitol concentrations from 0.4 M to 1.0M to induce hyper-osmotic stress and by varying the temperature of the thermo-jacketed pre-incubation chamber from 25 oC to 10 oC to impose sub-optimal temperature stress. Compared to controls (0.4 M sorbitol and 25 OC, the mesophyll protoplasts showed remarkable decrease in NaHCO3-dependent O2 evolution (indicator of photosynthetic carbon assimilation, under both hyper-osmotic (1.0 M sorbitol and sub-optimal temperature stress conditions (10 OC, while the decrease in rates of respiratory O2 uptake were marginal. The capacity of AOX pathway increased significantly in parallel to increase in intracellular pyruvate and reactive oxygen species (ROS levels under both hyper-osmotic stress and sub-optimal temperature stress under the background of saturating light. The ratio of redox couple (Malate/OAA related to malate valve increased in contrast to the ratio of redox couple (GSH/GSSG related to antioxidative system during hyper-osmotic stress. Nevertheless, the ratio of GSH/GSSG decreased in the presence of sub-optimal temperature, while the ratio of Malate/OAA showed no visible changes. Also, the redox ratios of pyridine nucleotides increased under hyper-osmotic (NADH/NAD and sub-optimal temperature (NADPH/NADP stresses, respectively. However, upon restriction of AOX pathway by using salicylhydroxamic acid (SHAM, the observed changes in NaHCO3 dependent O2 evolution, cellular ROS, redox ratios of Malate/OAA, NAD(PH/NAD(P and GSH/GSSG were further aggravated under stress conditions with concomitant modulations in NADP-MDH and antioxidant enzymes. Taken together, the

  4. An Alternative Procedure for the Glucose Oxidase Assay of Glucose as Applied to the Lactase Activity Assay (United States)

    Corbin Mullis, T.; Winge, Jeffery T.; Deal, S. Todd


    The glucose oxidase assay of glucose has been modified to eliminate the use of micropipets. The modification involves the use of disposable Pasteur pipets and a specified number of drops of each reagent. This simplified technique gives accurate and reproducible results.

  5. Teaching partner-focused questions to individuals who use augmentative and alternative communication to enhance their communicative competence. (United States)

    Light, J C; Binger, C; Agate, T L; Ramsay, K N


    A single-subject, multiple-probe experimental design was used to investigate the effect of instruction on the acquisition, generalization, and long-term maintenance of partner-focused questions (i.e., questions about communication partners and their experiences) by individuals who use augmentative and alternative communication (AAC). Six participants who had severe speech impairments and used AAC participated in the study; they ranged in age from 10 to 44 years, had a variety of disabilities, and used a range of AAC systems. Instruction used a least-to-most prompting hierarchy in real-world interactions and during simulations. All of the participants successfully learned to ask partner-focused questions spontaneously in social interactions; they required an average of approximately 6 hours of instruction (range: 3-11 hours). The participants generalized the use of partner-focused questions to new situations in the natural environment and maintained use of partner-focused questions at least 2 months postinstruction; one participant required some "booster" instructional sessions 4 weeks postinstruction to maintain her long-term use of partner-focused questions. The participants all reported high levels of satisfaction with the outcomes of the instructional program, as did their facilitators. Members of the general public, blind to the goals of the study, judged the majority of the participants to be more competent communicators after instruction.

  6. Structural studies of alternative oxidase (AOX) from moniliophthora perniciosa, the causal agent of witches' broom disease in cacao: a membrane-associated protein

    Energy Technology Data Exchange (ETDEWEB)

    Oliveira, J.F.; Prado, P.F.V.; Tiezzi, H.O.; Dias, S.M.G.; Ambrosio, A.L.B. [Laboratorio Nacional de Biociencias - LNBIO, Campinas, SP (Brazil); Thomazella, D.P.T.; Teixeira, P.J.P.L.; Pereira, G.A.G. [Universidade Estadual de Campinas (UNICAMP), SP (Brazil)


    Full text: Alternative oxidase (AOX) is a protein attached to the inner mitochondrial membrane that receives electrons directly from reduced ubiquinone and catalyzes the reduction of oxygen to water. AOX is a non-proton motive terminal quinol oxidase that enables cell respiration to continue even in the presence of inhibitors targeting the complexes of the respiratory chain. This protein is present in higher plants, pathogenic fungi and some parasites. The structural characterization of AOX becomes interesting due to its potential as a fungicide target. AOX is predicted to be a monotopic interfacial membrane protein interacting with a single leaflet of the lipid bilayer, rather than transmembrane. Amino acid sequence analysis reveals the presence of two conserved glutamate-histidine motifs, identifying it as a member of the diiron carboxylate protein family. The AOX model is defined by two pairs of helices forming a four helix bundle and an additional hydrophobic connecting sequence between the two helical pairs is proposed to act as the membrane anchoring region. In this work we aim at production, purification and crystallization of the AOX protein from M. perniciosa for further structural studies of this membrane-associated protein, by X-ray protein crystallography (author)

  7. Study of the effects of salicylic acid on soybean mitochondrial lipids and respiratory properties using the alternative oxidase as a stress-reporter protein. (United States)

    Matos, Ana Rita; Mendes, Ana Teresa; Scotti-Campos, Paula; Arrabaça, João Daniel


    Biotic and abiotic stresses can lead to modifications in the lipid composition of cell membranes. Although mitochondria appear to be implicated in stress responses, little is known about the membrane lipid changes that occur in these organelles in plants. Besides cytochrome c oxidase, plant mitochondria have an alternative oxidase (AOX) that accepts electrons directly from ubiquinol, dissipating energy as heat. AOX upregulation occurs under a variety of stresses and its induction by salicylic acid (SA) has been observed in different plant species. AOX was also suggested to be used as a functional marker for cell reprogramming under stress. In the present study, we have used etiolated soybean (Glycine max (L.) Merr. cv Cresir) seedlings to study the effects of SA treatment on the lipid composition and the respiratory properties of hypocotyl mitochondria. AOX expression was studied in detail, as a reporter protein, to evaluate whether modifications in mitochondrial energy metabolism were occurring. In mitochondria extracted from SA-treated seedlings, AOX capacity and protein contents increased. Both AOX1 and AOX2b transcripts accumulated in response to SA, but with different kinetics. A reduction in external NADH oxidation capacity was observed, whereas succinate respiration remained unchanged. The phospholipid composition of mitochondria remained similar in control and SA-treated plants, but a reduction in the relative amount of linolenic acid was observed in phosphatidylcholine, phosphatidylethanolamine and cardiolipin. The possible causes of the fatty acid modifications observed, and the implications for mitochondrial metabolism are discussed.

  8. Human retina-specific amine oxidase: genomic structure of the gene (AOC2), alternatively spliced variant, and mRNA expression in retina. (United States)

    Imamura, Y; Noda, S; Mashima, Y; Kudoh, J; Oguchi, Y; Shimizu, N


    Previously, we reported the isolation of cDNA for human retina-specific amine oxidase (RAO) and the expression of RAO exclusively in retina. Bacterial artificial chromosome clones containing the human RAO gene (AOC2) were mapped to human chromosome 17q21 (Imamura et al., 1997, Genomics 40: 277-283). Here, we report the complete genomic structure of the RAO gene, including 5' flanking sequence, and mRNA expression in retina. The human RAO gene spans 6 kb and is composed of four exons corresponding to the amino acid sequence 1-530, 530-598, 598-641, and 642-729 separated by three introns of 3000, 310, and 351 bp. Screening of a human retina cDNA library revealed the existence of an alternatively spliced cDNA variant with an additional 81 bp at the end of exon 2. The sizes of exons and the locations of exon/intron boundaries in the human RAO gene showed remarkable similarity to those of the human kidney diamine oxidase gene (AOC1). In situ hybridization revealed that mRNA coding for RAO is expressed preferentially in the ganglion cell layer of the mouse retina. We designed four sets of PCR primers to amplify four exons, which will be valuable for analyzing mutations in patients with ocular diseases affecting the retinal ganglion cell layer.

  9. Daucus carota L.--an old model for cell reprogramming gains new importance through a novel expansion pattern of alternative oxidase (AOX) genes. (United States)

    Costa, J H; Cardoso, H G; Campos, M D; Zavattieri, A; Frederico, A M; Fernandes de Melo, D; Arnholdt-Schmitt, B


    The paper highlights Daucus carota L. as an ideal model to complement plant stress research on Arabidopsis thaliana L. Recently, alternative oxidase (AOX) is discussed as functional marker candidate for cell reprogramming upon stress. Carrot is the most studied species for cell reprogramming and our current research reveals that it is the only one that has expanded both AOX sub-family genes. We point to recently published, but not discussed results on conserved differences in the vicinity of the most active functional site of AOX1 and AOX2, which indicate the importance of studying AOX sequence polymorphism, structure and functionality. Thus, stress-inducible experimental systems of D. carota are especially appropriate to bring research on stress tolerance a significant step forward.

  10. Anoxia pretreatment protects soybean cells against H(2)O(2)-induced cell death: possible involvement of peroxidases and of alternative oxidase. (United States)

    Amora, Y; Chevionb, M; Levinea, A


    Anoxia followed by reoxygenation causes extensive damage to cellular components through generation of reactive oxygen intermediates. We examined cellular responses to oxidative stress after anoxia in cultured soybean or human fibroblast cells. Anoxia pretreatment protected soybean but not fibroblasts against H(2)O(2) concentrations that induced programmed cell death in normoxic cells. H(2)O(2) removal in anoxia-pretreated soybean cultures was faster. Protection was associated with increased action of alternative oxidase (AOX) and peroxidases. AOX inhibitors abolished the protective effect, while induction of AOX protected normoxic cells against H(2)O(2). We propose that during anoxia, plant cells can prepare for reoxygenation injury by up-regulating their antioxidant capacity, and that AOX is involved in this process.

  11. Cloning and Expression of an Alternative Oxidase Gene from Lycopersicon esculentum%番茄交替氧化酶基因的克隆和表达

    Institute of Scientific and Technical Information of China (English)

    宋从凤; Broth wayne; 王金生; 胡晋生


    A full-length cDNA gene (LeAoxlau) was isolated from a cDNA library made from ripening fruit probing with alternative oxidase (AOX) gene fragments, obtained by degenerate primer PCR. Sequence analysis showed that LeAoxlau was 1 418 bp long and contained a 1 077-bp open reading frame encoding a about 40 kD precursor protein which is processed to a mature protein of 32 kD. Southern blot analysis suggested LeAoxlau is present as a single copy in the genome of tomato. RT-PCR analysis indicated LeAoxlau was expressed in roots, stems, leaves and cotyledons of tomato plants. A recombinant construct containing the open reading frame sequence of the LeAoxlau was transformed into Escherichia coli to express the alternative oxidase precursor protein.%利用简并PCR扩增产物做探针筛选番茄cDNA基因文库获得一个全长交替氧化酶cDNA基因LeAoxlau.经序列分析得出,该基因全长1 418bp,编码区序列长1 077 bp,编码约40 kD的前体蛋白.该蛋白在转运到线粒体时被加工成32kD的成熟蛋白.Southern印迹杂交分析结果显示该基因以单拷贝形式存在于番茄的基因组中RT-PCR显示,该基因在在番茄植株的根、茎、叶和子叶中表达.重组表达实验表明该基因能在大肠杆菌中表达.

  12. Alternative Approaches in Evaluating the EU SME Policy: Answers to the Question of Impact and Legitimization

    Directory of Open Access Journals (Sweden)

    Robert K. GRUENWALD


    Findings:  OECD  and  EU  evaluations  do  not  determine  causal  relationships  between funding allocation and effects. The evaluations of the KfW and the German Ministry of Economics  use  an  empirical  quantitative  approach  and  determine  direct  causal relations. In  order to fulfil the requirements of legitimizing functions  for  the  SME  policy,  it  is  recommended  to  further  develop  the  EU  funding policy  and  evaluation  according  to  the  “German  model”  both  in  terms  of  the institutional  framework  and  in  terms  of  the  evaluation  of  impacts  through  funding policy measures. Definition  of  minimum  requirements  and  alternative possibilities  for  EU  SME  policy  evaluation  in  order  to  close  the  legitimisation  gap between the allocation of tax money and impact proof (cost-benefit ratio.

  13. Open-ended questions: An alternative mode to assess the students' performance in concept development and use of scientific vocabulary (United States)

    Agairre-Ortiz, Maria


    Important reform movements across the nation recognize that improved science performance, language development, and alternative assessment are national priorities. This study examined the effect of two modes of alternative assessment on students' performance in in-depth concept development (IDC) and use of scientific vocabulary (VOC). The research questions asked for significant differences in students' performance in IDC and VOC across time regarding: (1) mode of assessment, (2) science achievement level, and classes. The study also investigated what are the: (1) students' opinions and feelings about the assessment modes (2) similarities and differences between OE-W and OE-D groups regarding the quality of IDC and VOC used. The sample of convenience included one hundred and four fifth grade LEP students randomly assigned to two groups. Students in both groups were asked to answer the same open-ended question at three assessment times during the study of a chapter on electricity and magnetism. Students in group one were asked to answer the question by writing paragraphs and students in group two answered the same question by making drawings. Results from a four-way repeated measures analysis showed that students in the OE-D mode, especially above-average performed better in in-depth concept development than those in the writing mode across time. The non-significant four-way interaction suggests that the differences in assessment mode across time are not influenced by science achievement level and the classes. The results suggest that although the OE-D mode in general yielded better scores, both modes of assessment could be used to assess Spanish-dominant LEP students' conceptual development and scientific vocabulary use. In general, students expressed preference for OE-D because they felt more confident and comfortable answering questions by drawing. Most of the statistical results were supported by the qualitative analysis for both dependent variables. Small size

  14. Teaching children who use augmentative and alternative communication to ask inverted yes/no questions using aided modeling. (United States)

    Kent-Walsh, Jennifer; Binger, Cathy; Buchanan, Carolyn


    This study investigated the effects of a direct intervention program involving aided modeling and the presentation of contrastive targets on the aided production of inverted yes/no questions and possible generalization to other sentence types by children using augmentative and alternative communication (AAC). A single-case, multiple-probe, experimental design across participants was used to evaluate the effects of the instructional program with 3 children who had motor speech disorders and used AAC (ages 4;10 [years;months], 6;2, and 4;9). The treatment involved aided modeling of treatment and contrastive targets through concentrated modeling and interactive play activities. Direct treatment outcomes were examined by measuring the accuracy of producing inverted yes/no questions and to be declaratives through probes. All 3 participants showed a direct treatment effect, producing a greater number of inverted yes/no questions and to be declaratives within the probes following treatment compared with before treatment. All 3 participants evidenced some generalization to novel sentences. Results provide initial evidence that instruction involving aided modeling with contrastive targets holds promise in targeting specific linguistic rules with children using AAC. Patterns of generalization may depend on participants' specific language deficits and acquisition patterns during intervention.

  15. Testing alternative explanations for common responses to conceptual questions: An example in the context of center of mass (United States)

    Heron, Paula R. L.


    In physics education research it has been common to interpret student errors on conceptual questions in topic-specific ways, rather than in terms of general perceptual or reasoning difficulties. This paper examines two alternative accounts for responses to questions related to the concept of center of mass. In one account, difficulties are said to be perceptual in nature; in the other, difficulties are said to be tightly linked to the concepts in question. Hypotheses derived from the former perspective are tested in studies conducted among university students in introductory physics courses. The results do not provide strong support for the perceptual hypothesis; in fact, there is evidence that performance on perception tasks may be influenced by subjects' ideas about the physical scenario. While the results do not provide general support for one perspective versus the other, the paper serves as an illustration of the type of investigation needed to develop the kind of rich representation of student thinking that will allow instructional resources to be most effectively targeted.

  16. Induction of mitochondrial alternative oxidase in response to a cell signal pathway down-regulating the cytochrome pathway prevents programmed cell death. (United States)

    Vanlerberghe, Greg C; Robson, Christine A; Yip, Justine Y H


    Treatment of tobacco (Nicotiana tabacum L. cv Petit Havana SR1) cells with cysteine (Cys) triggers a signal pathway culminating in a large loss of mitochondrial cytochrome (cyt) pathway capacity. This down-regulation of the cyt path likely requires events outside the mitochondrion and is effectively blocked by cantharidin or endothall, indicating that protein dephosphorylation is one critical process involved. Generation of reactive oxygen species, cytosolic protein synthesis, and Ca(2+) flux from organelles also appear to be involved. Accompanying the loss of cyt path is a large induction of alternative oxidase (AOX) protein and capacity. Induction of AOX allows the cells to maintain high rates of respiration, indicating that the lesion triggered by Cys is in the cyt path downstream of ubiquinone. Consistent with this, transgenic (AS8) cells unable to induce AOX (due to the presence of an antisense transgene) lose all respiratory capacity upon Cys treatment. This initiates in AS8 a programmed cell death pathway, as evidenced by the accumulation of oligonucleosomal fragments of DNA as the culture dies. Alternatively, wild-type cells remain viable and eventually recover their cyt path. Induction of AOX in response to a chemical inhibition of the cyt path (by antimycin A) is also dependent upon protein dephosphorylation and the generation of reactive oxygen species. Common events required for both down-regulation of the cyt path and induction of AOX may represent a mechanism to coordinate the biogenesis of these two electron transport paths. Such coordinate regulation may be necessary, not only to satisfy metabolic demands, but also to modulate the initiation of a programmed cell death pathway responsive to mitochondrial respiratory status.

  17. Disruption of the mitochondrial alternative oxidase (AOX) and uncoupling protein (UCP) alters rates of foliar nitrate and carbon assimilation in Arabidopsis thaliana. (United States)

    Gandin, Anthony; Denysyuk, Mykhaylo; Cousins, Asaph B


    Under high light, the rates of photosynthetic CO2 assimilation can be influenced by reductant consumed by both foliar nitrate assimilation and mitochondrial alternative electron transport (mAET). Additionally, nitrate assimilation is dependent on reductant and carbon skeletons generated from both the chloroplast and mitochondria. However, it remains unclear how nitrate assimilation and mAET coordinate and contribute to photosynthesis. Here, hydroponically grown Arabidopsis thaliana T-DNA insertional mutants for alternative oxidase (AOX1A) and uncoupling protein (UCP1) fed either NO3 (-) or NH4 (+) were used to determine (i) the response of NO3 (-) uptake and assimilation to the disruption of mAET, and (ii) the interaction of N source (NO3 (-) versus NH4 (+)) and mAET on photosynthetic CO2 assimilation and electron transport. The results showed that foliar NO3 (-) assimilation was enhanced in both aox1a and ucp1 compared with the wild-type, suggesting that foliar NO3 (-) assimilation is probably driven by a decreased capacity of mAET and an increase in reductant within the cytosol. Wild-type plants had also higher rates of net CO2 assimilation (A net) and quantum yield of PSII (ϕPSII) under NO3 (-) feeding compared with NH4 (+) feeding. Additionally, under NO3 (-) feeding, A net and ϕPSII were decreased in aox1a and ucp1 compared with the wild type; however, under NH4 (+) they were not significantly different between genotypes. This indicates that NO3 (-) assimilation and mAET are both important to maintain optimal rates of photosynthesis, probably in regulating reductant accumulation and over-reduction of the chloroplastic electron transport chain. These results highlight the importance of mAET in partitioning energy between foliar nitrogen and carbon assimilation.

  18. Transgenic plant cells lacking mitochondrial alternative oxidase have increased susceptibility to mitochondria-dependent and -independent pathways of programmed cell death. (United States)

    Robson, Christine A; Vanlerberghe, Greg C


    The plant mitochondrial electron transport chain is branched such that electrons at ubiquinol can be diverted to oxygen via the alternative oxidase (AOX). This pathway does not contribute to ATP synthesis but can dampen the mitochondrial generation of reactive oxygen species. Here, we establish that transgenic tobacco (Nicotiana tabacum L. cv Petit Havana SR1) cells lacking AOX (AS8 cells) show increased susceptibility to three different death-inducing compounds (H(2)O(2), salicylic acid [SA], and the protein phosphatase inhibitor cantharidin) in comparison with wild-type cells. The timing and extent of AS8 cell death are very similar among the three treatments and, in each case, are accompanied by the accumulation of oligonucleosomal fragments of DNA, indicative of programmed cell death. Death induced by H(2)O(2) or SA occurs by a mitochondria-dependent pathway characterized by cytochrome c release from the mitochondrion. Conversely, death induced by cantharidin occurs by a pathway without any obvious mitochondrial involvement. The ability of AOX to attenuate these death pathways may relate to its ability to maintain mitochondrial function after insult with a death-inducing compound or may relate to its ability to prevent chronic oxidative stress within the mitochondrion. In support of the latter, long-term treatment of AS8 cells with an antioxidant compound increased the resistance of AS8 cells to SA- or cantharidin-induced death. The results indicate that plants maintain both mitochondria-dependent and -independent pathways of programmed cell death and that AOX may act as an important mitochondrial "survival protein" against such death.

  19. Mitochondrial alternative oxidase acts to dampen the generation of active oxygen species during a period of rapid respiration induced to support a high rate of nutrient uptake. (United States)

    Yip, Justine Y. H.; Vanlerberghe, Greg C.


    When wild type (wt) tobacco (Nicotiana tabacum L. cv. Petit Havana SR1) suspension cells were grown under phosphate (P) limitation, they contained large amounts of mitochondrial alternative oxidase (AOX). When these cells were resupplied with P, there was a large, immediate and sustained stimulation of respiration to support a period of rapid P uptake. Two lines of evidence suggest that the abundant level of AOX present in wt cells contributed to this stimulated rate of respiration. First, when P-limited transgenic antisense tobacco cells (AS8) lacking AOX were resupplied with P, the stimulation of respiration was much less dramatic even though these cells displayed similar rates of P uptake. Second, while the stimulated rate of respiration in AS8 cells was insensitive (as expected) to the AOX inhibitor n-propyl gallate (nPG), much of the stimulated rate of respiration in wt cells could be inhibited by nPG. Given the non-phosphorylating nature of AOX respiration, wt cells required higher rates of electron transport to O2 than AS8 cells to support similar rates of P uptake. The utilization of AOX by wt cells during P uptake was apparently not occurring because the cytochrome (Cyt) pathway alone could not fully support the rate of P uptake, as the respiration of cells lacking AOX (either untreated AS8 cells or wt cells treated with nPG) supported similar rates of P uptake as wt cells with abundant AOX. Rather, we provide in vivo evidence that the utilization of AOX during the period of high respiration supporting P uptake was to dampen the mitochondrial generation of active oxygen species (AOS).

  20. Stress-induced co-expression of two alternative oxidase (VuAox1 and 2b) genes in Vigna unguiculata. (United States)

    Costa, José Hélio; Mota, Erika Freitas; Cambursano, Mariana Virginia; Lauxmann, Martin Alexander; de Oliveira, Luciana Maia Nogueira; Silva Lima, Maria da Guia; Orellano, Elena Graciela; Fernandes de Melo, Dirce


    Cowpea (Vigna unguiculata) alternative oxidase is encoded by a small multigene family (Aox1, 2a and 2b) that is orthologous to the soybean Aox family. Like most of the identified Aox genes in plants, VuAox1 and VuAox2 consist of 4 exons interrupted by 3 introns. Alignment of the orthologous Aox genes revealed high identity of exons and intron variability, which is more prevalent in Aox1. In order to determine Aox gene expression in V. unguiculata, a steady-state analysis of transcripts involved in seed development (flowers, pods and dry seeds) and germination (soaked seeds) was performed and systemic co-expression of VuAox1 and VuAox2b was observed during germination. The analysis of Aox transcripts in leaves from seedlings under different stress conditions (cold, PEG, salicylate and H2O2 revealed stress-induced co-expression of both VuAox genes. Transcripts of VuAox2a and 2b were detected in all control seedlings, which was not the case for VuAox1 mRNA. Estimation of the primary transcript lengths of V. unguiculata and soybean Aox genes showed an intron length reduction for VuAox1 and 2b, suggesting that the two genes have converged in transcribed sequence length. Indeed, a bioinformatics analysis of VuAox1 and 2b promoters revealed a conserved region related to a cis-element that is responsive to oxidative stress. Taken together, the data provide evidence for co-expression of Aox1 and Aox2b in response to stress and also during the early phase of seed germination. The dual nature of VuAox2b expression (constitutive and induced) suggests that the constitutive Aox2b gene of V. unguiculata has acquired inducible regulatory elements.

  1. Big Bang, inflation, standard Physics… and the potentialities of new Physics and alternative cosmologies. Present statuts of observational and experimental Cosmology. Open questions and potentialities of alternative cosmologies (United States)

    Gonzalez-Mestres, Luis


    A year ago, we wrote [1] that the field of Cosmology was undergoing a positive and constructive crisis. The possible development of more direct links between the Mathematical Physics aspects of cosmological patterns and the interpretation of experimental and observational results was particularly emphasized. Controversies on inflation are not really new, but in any case inflation is not required in pre-Big Bang models and the validity of the standard Big Bang + inflation + ΛCDM pattern has not by now been demonstrated by data. Planck has even explicitly reported the existence of "anomalies". Remembering the far-reaching work of Yoichiro Nambu published in 1959-61, it seems legitimate to underline the need for a cross-disciplinary approach in the presence of deep, unsolved theoretical problems concerning new domains of matter properties and of the physical world. The physics of a possible preonic vacuum and the associated cosmology constitute one of these domains. If the vacuum is made of superluminal preons (superbradyons), and if standard particles are vacuum excitations, how to build a suitable theory to describe the internal structure of such a vacuum at both local and cosmic level? Experimental programs (South Pole, Atacama, AUGER, Telescope Array…) and observational ones (Planck, JEM-EUSO…) devoted to the study of cosmic microwave background radiation (CMB) and of ultra-high energy cosmic rays (UHECR) are crucial to elucidate such theoretical interrogations and guide new phenomenological developments. Together with a brief review of the observational and experimental situation, we also examine the main present theoretical and phenomenological problems and point out the role new physics and alternative cosmologies can potentially play. The need for data analyses less focused a priori on the standard models of Particle Physics and Cosmology is emphasized in this discussion. An example of a new approach to both fields is provided by the pre-Big Bang pattern

  2. The terminal oxidases of Paracoccus denitrificans


    de Gier, Jan-Willem L.; Lübben, Mathias; Reijnders, Willem N.M.; Tipker, Corinne A.; Slotboom, Dirk-Jan; Van Spanning, Rob J. M.; Stouthamer, Adriaan H.; van der Oost, John


    Three distinct types of terminal oxidases participate in the aerobic respiratory pathways of Paracoccus denitrificans. Two alternative genes encoding subunit I of the aa3-type cytochrome c oxidase have been isolated before, namely ctaDI and ctaDII. Each of these genes can be expressed separately to complement a double mutant (ΔctaDI, ΔctaDII), indicating that they are isoforms of subunit I of the aa3-type oxidase. The genomic locus of a quinol oxidase has been isolated: cyoABC. This protohaem...

  3. Evidence-based practice method of integrative Chinese and Western medicine based on literature retrieval through PICO question and complementary and alternative medicine topics. (United States)

    Yan, Xiu-feng; Ni, Qing; Wei, Jun-ping; Xu, Hao


    An evidence-based practice method according to literature retrieval through PICO (Patients, Intervention, Comparison, Outcome) questions and complementary and alternative medicine (CAM) topics, which can obtain helpful evidence for guiding clinical practice, was introduced with a practical example in this paper. The knowledge of diseases and Western medicine treatment can be acquired by literature retrieval through PICO question, while searching by CAM topics may provide evidence for Chinese medicine (CM). Thus the author held that literature retrieval through both PICO question and CAM topics was an ideal evidence-based practice method for integrative Chinese and Western medicine (ICWM). However, since the standard in CM evidence hierarchy is still under study, the value of the CAM thematic retrieval method remains very limited. In the future, studies on the definition and hierarchy of CM evidences and the herb-drug interaction between Western and Chinese medicine during a combination therapy should be strengthened to improve the status of ICWM evidence-based practice.

  4. Deep boreholes. An alternative for final disposal of spent nuclear fuel? Report from KASAM's question-and-answer session on 14-15 March 2007

    Energy Technology Data Exchange (ETDEWEB)


    On 14-15 March 2007, KASAM held a hearing for the purpose of thoroughly examining deep boreholes as a method for the final disposal of spent nuclear fuel. Some of the questions that were raised were: What are the technical, geological and hydrological premises and possibilities? What are the risks from different viewpoints and what values underlie different views of the potential and suitability of deep boreholes? This report is a summary of the seminar. KASAM has made a selection of contributions and questions from the debate that took place on the basis of their relevance to the purpose of the seminar. The report generally follows the chronological lecture-and debate format of the seminar, but has been edited according to different issues rather than according to when different persons spoke. Chapter 2 describes a number of premises and criteria in the Environmental Code's and the Nuclear Activities Act's requirements on alternatives reporting. The chapter also contains a description of what the deep borehole concept entails and a discussion of the geoscientific premises. In addition, the chapter describes how different values can influence the choice of final disposal method. Chapters 3-6 describe and discuss technology and long-term safety, the viewpoints of the supervisory authorities on deep boreholes and safety philosophy via lectures followed by questions by KASAM's questioners and the audience. On the evening of 14 March, representatives of the seven parliamentary parties discussed their preparations and standpoints for an upcoming national debate on the final disposal of nuclear waste. This discussion is also reproduced in the report as Chapter 7. The main points from a concluding panel debate and discussion are presented in Chapter 8. In conclusion, Chapter 9 contains some reflections on various arguments proffered during the question-and-answer session, questions on which agreement seems to exist, and where there are differences of

  5. An antibody against a conserved C-terminal consensus motif from plant alternative oxidase (AOX) isoforms 1 and 2 label plastids in the explosive dwarf mistletoe (Arceuthobium americanum, Santalaceae) fruit exocarp. (United States)

    Ross Friedman, Cynthia; Ross, Bradford N; Martens, Garnet D


    Dwarf mistletoes, genus Arceuthobium (Santalaceae), are parasitic angiosperms that spread their seeds by an explosive process. As gentle heating triggers discharge in the lab, we wondered if thermogenesis (endogenous heat production) is associated with dispersal. Thermogenesis occurs in many plants and is enabled by mitochondrial alternative oxidase (AOX) activity. The purpose of this study was to probe Arceuthobium americanum fruit (including seed tissues) collected over a 10-week period with an anti-AOX antibody/gold-labeled secondary antibody to determine if AOX could be localized in situ, and if so, quantitatively assess whether label distribution changed during development; immunochemical results were evaluated with Western blotting. No label could be detected in the mitochondria of any fruit or seed tissue, but was observed in fruit exocarp plastids of samples collected in the last 2 weeks of study; plastids collected in week 10 had significantly more label than week 9 (p = 0.002). Western blotting of whole fruit and mitochondrial proteins revealed a signal at 30-36 kD, suggestive of AOX, while blots of whole fruit (but not mitochondrial fraction) proteins showed a second band at 40-45 kD, in agreement with plastid terminal oxidases (PTOXs). AOX enzymes are likely present in the A. americanum fruit, even though they were not labeled in mitochondria. The results strongly indicate that the anti-AOX antibody was labeling PTOX in plastids, probably at a C-terminal region conserved in both enzymes. PTOX in plastids may be involved in fruit ripening, although a role for PTOX in thermogenesis cannot be eliminated.

  6. 西瓜交替氧化酶AOX2基因的克隆与分析%Cloning and analysis of the alternative oxidase-2(AOX2) gene in watermelon

    Institute of Scientific and Technical Information of China (English)

    李严曼; 朱磊; 杨景华; 张明方


    In order to study the possible functions of the alternative oxidase gene family in watermelon plants,high-quality RNA was extracted from cold-tolerant watermelon(Citrullus lanatus) germplasm IVSM No.9 and then cDNA was obtained through reverse transcription kit.A cDNA fragment of alternative oxidase was amplified using degenerate oligonucleotide primers designed on the basis of the conserved domain of alternative oxidase genes in other plants.According to the cDNA fragment sequence,the 5' and 3' end fragment sequences were obtained using RACE technology respectively.The full length cDNA of ClAOX2 was obtained by DNAMAN5.22 from the above three sequences.The cDNA fragment exhibited high homology to the alternative oxidase-2(AOX2) of other species and could be referred to as ClAOX2.Then the full lengths of cDNA and DNA were obtained by PCR according to the known cDNA sequence.The full-length cDNA of ClAOX2 was 1 263 bp in size with a 1050 bp open reading frame(ORF).The full-length DNA was 1 983 bp which has consisted of four exons interrupted by three introns.The accession number of ClAOX2 in GenBank database is ADD84880.Real-time PCR showed that the ClAOX2 gene expressed in root,stem,leaf,flower,and fruit of watermelon seedling and significantly higher in the fruit.These results demonstrated that ClAOX2 may play a role during fruit development.%为了研究西瓜交替氧化酶基因家族在西瓜植株中可能发挥的功能,以西瓜(Citrullus lanatus)耐冷种质IVSM9为材料,根据植物不同物种交替氧化酶基因核苷酸保守区序列设计兼并引物,得到西瓜交替氧化酶(alterna-tive oxidase)AOX基因的中间片段。在已知序列的基础上,分别设计5’和3’末端扩增的特异引物,采用cDNA末端快速扩增(RACE)的方法得到西瓜交替氧化酶基因的5’端和3’端序列,用DNAMAN5.22软件对3个序列进行拼接和分析,获得全长序列。得到的基因片段同其他物种AOX基因家族中的alternative

  7. Induction of the AOX1D Isoform of Alternative Oxidase in A. thaliana T-DNA Insertion Lines Lacking Isoform AOX1A Is Insufficient to Optimize Photosynthesis when Treated with Antimycin A

    Institute of Scientific and Technical Information of China (English)

    Inga Strodtkotter; Kollipara Padmasree; Challabathula Dinakar; Birgit Speth; Pamela S. Niazi; Joanna Wojtera; Ingo Voss; Phuc Thi Do; Adriano Nunes-Nesi; Alisdair R. Fernie; Vera Linke; Agepati S. Raghavendra; Renate Scheibe


    Plant respiration is characterized by two pathways for electron transfer to O2, namely the cytochrome path-way (CP) that is linked to ATP production, and the alternative pathway (AP), where electrons from ubiquinol are directly transferred to O2 via an alternative oxidase (AOX) without concomitant ATP production. This latter pathway is well suited to dispose of excess electrons in the light, leading to optimized photosynthetic performance. We have characterized T-DNA-insertion mutant lines of Arabidopsis thaliana that do not express the major isoform, AOXIA. In standard growth conditions, these plants did not show any phenotype, but restriction of electron flow through CP by antimycin A, which induces AOX1A expression in the wild-type, led to an increased expression of AOX1D in leaves of the aox1a-knockout mutant. Despite the increased presence of the AOX1D isoform in the mutant, antimycin A caused inhibition of photosyn-thesis, increased ROS, and ultimately resulted in amplified membrane leakage and necrosis when compared to the wild-type, which was only marginally affected by the inhibitor. It thus appears that AOX1D was unable to fully compensate for the loss of AOX1A when electron flow via the CP is restricted. A combination of inhibition studies, coupled to metabolite profiling and targeted expression analysis of the P-protein of glycine decarboxylase complex (GDC), suggests that the aox1a mutants attempt to increase their capacity for photorespiration. However, given their deficiency, it is intriguing that increase in expression neither of AOX1D nor of GDC could fully compensate for the lack of AOX1A to optimize pho-tosynthesis when treated with antimycin A. We suggest that the aox1a mutants can further be used to substantiate the current models concerning the influence of mitochondrial redox on photosynthetic performance and gene expression.

  8. Mitochondrial cytochrome c oxidase deficiency. (United States)

    Rak, Malgorzata; Bénit, Paule; Chrétien, Dominique; Bouchereau, Juliette; Schiff, Manuel; El-Khoury, Riyad; Tzagoloff, Alexander; Rustin, Pierre


    As with other mitochondrial respiratory chain components, marked clinical and genetic heterogeneity is observed in patients with a cytochrome c oxidase deficiency. This constitutes a considerable diagnostic challenge and raises a number of puzzling questions. So far, pathological mutations have been reported in more than 30 genes, in both mitochondrial and nuclear DNA, affecting either structural subunits of the enzyme or proteins involved in its biogenesis. In this review, we discuss the possible causes of the discrepancy between the spectacular advances made in the identification of the molecular bases of cytochrome oxidase deficiency and the lack of any efficient treatment in diseases resulting from such deficiencies. This brings back many unsolved questions related to the frequent delay of clinical manifestation, variable course and severity, and tissue-involvement often associated with these diseases. In this context, we stress the importance of studying different models of these diseases, but also discuss the limitations encountered in most available disease models. In the future, with the possible exception of replacement therapy using genes, cells or organs, a better understanding of underlying mechanism(s) of these mitochondrial diseases is presumably required to develop efficient therapy.

  9. Transgenic cassava lines carrying heterologous alternative oxidase ...

    African Journals Online (AJOL)

    To screen positive lines for gene function, leaf lobes from two transgenic lines with a line carrying an empty vector and the wild type were subjected to somatic embryogenesis (SE), a known oxidative ... African Journal of Biotechnology Vol.

  10. The six most essential questions in psychiatric diagnosis: a pluralogue part 3: issues of utility and alternative approaches in psychiatric diagnosis

    Directory of Open Access Journals (Sweden)

    Phillips James


    Full Text Available Abstract In face of the multiple controversies surrounding the DSM process in general and the development of DSM-5 in particular, we have organized a discussion around what we consider six essential questions in further work on the DSM. The six questions involve: 1 the nature of a mental disorder; 2 the definition of mental disorder; 3 the issue of whether, in the current state of psychiatric science, DSM-5 should assume a cautious, conservative posture or an assertive, transformative posture; 4 the role of pragmatic considerations in the construction of DSM-5; 5 the issue of utility of the DSM – whether DSM-III and IV have been designed more for clinicians or researchers, and how this conflict should be dealt with in the new manual; and 6 the possibility and advisability, given all the problems with DSM-III and IV, of designing a different diagnostic system. Part 1 of this article took up the first two questions. Part 2 took up the second two questions. Part 3 now deals with Questions 5 & 6. Question 5 confronts the issue of utility, whether the manual design of DSM-III and IV favors clinicians or researchers, and what that means for DSM-5. Our final question, Question 6, takes up a concluding issue, whether the acknowledged problems with the earlier DSMs warrants a significant overhaul of DSM-5 and future manuals. As in Parts 1 & 2 of this article, the general introduction, as well as the introductions and conclusions for the specific questions, are written by James Phillips, and the responses to commentaries are written by Allen Frances.

  11. Questioning Techniques. (United States)

    Patterson, Michelle


    Questions are so much a part of the classroom routine and they should stimulate learning and thinking. Introduces the Questioning and Understanding to Improve Learning and Thinking (QUILT) method which incorporates Bloom's Taxonomy and wait time. (ASK)

  12. Cucumber Seedling Indoleacetaldehyde Oxidase 1 (United States)

    Bower, Peter J.; Brown, Hugh M.; Purves, William K.


    Extracts of light-grown Cucumis sativus L. seedlings catalyzed the oxidation of indole-3-acetaldehyde to indole-3-acetic acid. No added cofactors were required. Inhibitor studies indicated that the enzyme is a metalloflavoprotein. While indole-3-aldehyde, benzaldehyde, and phenylacetaldehyde partially inhibited the oxidation of indole-3-acetaldehyde, suggesting that they may serve as alternative substrates, it is proposed that indoleacetaldehyde is the major substrate in vivo. 2,4-Dichlorophenoxyacetic acid strongly inhibited the indoleacetaldehyde oxidase activity, and it is proposed that this enzyme may be subject in vivo to feedback inhibition by indole-3-acetic acid. The enzyme was activated by brief heating or by treatment with mercaptoethanol. PMID:16660220

  13. Curiosity Questions (United States)

    Nelsen, Jane; DeLorenzo, Chip


    Have you ever found yourself lecturing a child, with the best of intentions, in an attempt to help him or her learn a lesson or process a situation in a manner that you feel will be productive? Curiosity questions, which the authors also call What and How questions, help children process an experience, event, or natural consequence so that they…

  14. Isolated sulfite oxidase deficiency. (United States)

    Relinque, B; Bardallo, L; Granero, M; Jiménez, P J; Luna, S


    Sulfite oxidase deficiency is an uncommon metabolic disease. Only few cases of its isolated form have been reported in the literature. We report a case of severe neonatal onset. A newborn baby of 41 weeks gestational age, weighted at birth of 3240 grams and had an Apgar score of 6-10-10. Fifty-three hours after being born, the baby started with seizures that were refractory to antiepileptic treatment. Brain function was monitored using a-EEG. Laboratory and imaging tests were performed. All of them were consistent with sulfite oxidase deficiency. The diagnosis was confirmed by genetic testing. We highlight the importance of this disease as part of the differential diagnosis of seizures during the neonatal period, as well as the importance of the therapeutic support based on dietary restrictions. It's also remarkable the possibility of prenatal diagnosis by quantifying enzyme activity and it's also possible carrying out DNA mutational analysis.

  15. Alternative metrics (United States)


    As the old 'publish or perish' adage is brought into question, additional research-impact indices, known as altmetrics, are offering new evaluation alternatives. But such metrics may need to adjust to the evolution of science publishing.


    Directory of Open Access Journals (Sweden)



    Full Text Available INTRODUCTION: Surveillance for risk factors of heart diseases, like increased blood sugar, cholesterol becomes difficult in places with inadequate lab facilities due to difficulty in sample collection, transportation and processing. Feasibility of using dried blood/seru m spots in such situations can be thought of as an alternative to fresh serum total cholesterol, as sample collection does not require much expertise. AIM: The aim of this study is to determine whether dried blood and serum spots can be used as an alternat ive to fresh serum for estimation of total cholesterol in field studies. METHOD S : Fresh serum from 100 subjects selected randomly was used for estimation of total cholesterol. Four spots each of blood and serum from these samples were made on Whatman paper 3, out of which one spot from each were analysed on the day of collection and rest were kept in resealable bags to be analysed on day 7, 15 and 30. The correlation between the dried blood/serum spot values with the fresh serum total cholesterol values was examined. RESULTS: A significant correlation with fresh serum total cholesterol values (p value<0.01 was seen for dried serum spots stored up to day 30(0.747 to 0.942 and for dried blood spots stored up to day 15(0.598 to 0.949. CONCLUSIONS: Total chol esterol estimated by dried blood spot up to 15 days and dried serum spots stored up to 30 days can be considered as an alternative to fresh serum for estimation of serum total cholesterol.

  17. Unproven (questionable) cancer therapies. (United States)

    Brigden, M L


    More than half of all cancer patients use some form of alternative treatment during the course of their illness. Alternative therapies are often started early in patients' illness, and their use is frequently not acknowledged to health care professionals. Some alternative therapies are harmful, and their promoters may be fraudulent. Persons who try alternative cancer therapies may not be poorly educated but may ultimately abandon conventional treatment. Recent attention has focused on aspects of questionable therapies that make these treatments attractive to patients and that may be perceived as being deficient in the practice of conventional health care professionals. Physicians with patients with cancer should always make sure that unproven therapies are discussed early in the therapeutic relationship. They should also attempt to be aware of alternative therapies that are in vogue in their particular geographic area.

  18. Thousand Questions

    DEFF Research Database (Denmark)

    Soon, Winnie; Pritchard, Helen


    In this work the network asks “If I wrote you a love letter would you write back?” Like the love letters which appeared mysteriously on the noticeboards of Manchester University’s Computer Department in the 1950s, thousands of texts circulate as computational processes perform the questions...... (perhaps as an expanded Turing test) on its listeners. These questions are extracted in real-time from Twitter with the keyword search of the ‘?’ symbol to create a spatio-temporal experience. The computerized voice the audience hears is a collective one, an entanglement of humans and non...

  19. Critical Questions. (United States)

    Worthy, Jo; Hoffman, James V.


    Offers responses from four readers of this journal, all reading and/or classroom teachers, to a question posed by another teacher: whether children who have had limited literacy experiences should start reading in whole-language readers and/or trade books or whether they should start in controlled-vocabulary preprimers. (SR)

  20. Thousand Questions

    DEFF Research Database (Denmark)


    (perhaps as an expanded Turing test) on its listeners. These questions are extracted in real-time from Twitter with the keyword search of the ‘?’ symbol to create a spatio-temporal experience. The computerized voice the audience hears is a collective one, an entanglement of humans and non...

  1. My Questions

    Institute of Scientific and Technical Information of China (English)


    I have many questions.I think hard and cannot find the answers.I wonder what the world is in reality.Is there any answer to the mystery of the world?I ask what it is like to be outside the earth.Are there any people on other planets1?How many planets are there in the universe?I wonder a lot about this and that.

  2. Structural and functional analysis of aa3-type and cbb3-type cytochrome c oxidases of Paracoccus denitrificans reveals significant differences in proton-pump design


    de Gier, Jan-Willem L.; Schepper, Mike; Reijnders, Willem N.M.; Dyck, Stef J. van; Slotboom, Dirk Jan; Warne, Antony; Saraste, Matti; Krab, Klaas; Finel, Moshe; Stouthamer, Adriaan H.; Van Spanning, Rob J. M.; van der Oost, John


    In Paracoccus denitrificans the aa3-type cytochrome c oxidase and the bb3-type quinol oxidase have previously been characterized in detail, both biochemically and genetically. Here we report on the isolation of a genomic locus that harbours the gene cluster ccoNOQP, and demonstrate that it encodes an alternative cbb3-type cytochrome c oxidase. This oxidase has previously been shown to be specifically induced at low oxygen tensions, suggesting that its expression is controlled by an oxygen-sen...

  3. Lysyl oxidase in colorectal cancer. (United States)

    Cox, Thomas R; Erler, Janine T


    Colorectal cancer is the third most prevalent form of cancer worldwide and fourth-leading cause of cancer-related mortality, leading to ~600,000 deaths annually, predominantly affecting the developed world. Lysyl oxidase is a secreted, extracellular matrix-modifying enzyme previously suggested to act as a tumor suppressor in colorectal cancer. However, emerging evidence has rapidly implicated lysyl oxidase in promoting metastasis of solid tumors and in particular colorectal cancer at multiple stages, affecting tumor cell proliferation, invasion, and angiogenesis. This emerging research has stimulated significant interest in lysyl oxidase as a strong candidate for developing and deploying inhibitors as functional efficacious cancer therapeutics. In this review, we discuss the rapidly expanding body of knowledge concerning lysyl oxidase in solid tumor progression, highlighting recent advancements in the field of colorectal cancer.

  4. Lysyl oxidase in colorectal cancer

    DEFF Research Database (Denmark)

    Cox, Thomas R; Erler, Janine T


    Colorectal cancer is the third most prevalent form of cancer worldwide and fourth-leading cause of cancer-related mortality, leading to ~600,000 deaths annually, predominantly affecting the developed world. Lysyl oxidase is a secreted, extracellular matrix-modifying enzyme previously suggested...... to act as a tumor suppressor in colorectal cancer. However, emerging evidence has rapidly implicated lysyl oxidase in promoting metastasis of solid tumors and in particular colorectal cancer at multiple stages, affecting tumor cell proliferation, invasion, and angiogenesis. This emerging research has...... advancements in the field of colorectal cancer....

  5. Peroxisomal Polyamine Oxidase and NADPH-Oxidase cross-talk for ROS homeostasis which affects respiration rate in Arabidopsis thaliana

    Directory of Open Access Journals (Sweden)

    Efthimios A. Andronis


    Full Text Available Homeostasis of reactive oxygen species (ROS in the intracellular compartments is of critical importance as ROS have been linked with nearly all cellular processes and more importantly with diseases and aging. PAs are nitrogenous molecules with an evolutionary conserved role in the regulation of metabolic and energetic status of cells. Recent evidence also suggests that polyamines (PA are major regulators of ROS homeostasis. In Arabidopsis the backconversion of the PAs spermidine (Spd and spermine (Spm to putrescine (Put and Spd, respectively is catalyzed by two peroxisomal PA oxidases (AtPAO. However, the physiological role of this pathway remains largely elusive. Here we explore the role of peroxisomal PA backconversion and in particular that catalyzed by the highly expressed AtPAO3 in the regulation of ROS homeostasis and mitochondrial respiratory burst. Exogenous PAs exert an NADPH-oxidase dependent stimulation of oxygen consumption, with Spd exerting the strongest effect. This increase is attenuated by treatment with the NADPH-oxidase blocker diphenyleneiodonium iodide (DPI. Loss-of-function of AtPAO3 gene results to increased NADPH-oxidase-dependent production of superoxide anions (O2.-, but not H2O2, which activate the mitochondrial alternative oxidase pathway (AOX. On the contrary, overexpression of AtPAO3 results to an increased but balanced production of both H2O2 and O2.-. These results suggest that the ratio of O2.-/H2O2 regulates respiratory chain in mitochondria, with PA-dependent production of O2.- by NADPH-oxidase tilting the balance of electron transfer chain in favor of the AOX pathway. In addition, AtPAO3 seems to be an important component in the regulating module of ROS homeostasis, while a conserved role for PA backconversion and ROS across kingdoms is discussed.

  6. Flavoprotein oxidases : classification and applications

    NARCIS (Netherlands)

    Dijkman, Willem P.; de Gonzalo, Gonzalo; Mattevi, Andrea; Fraaije, Marco W.


    This review provides an overview of oxidases that utilise a flavin cofactor for catalysis. This class of oxidative flavoenzymes has shown to harbour a large number of biotechnologically interesting enzymes. Applications range from their use as biocatalysts for the synthesis of pharmaceutical compoun

  7. Disruption of the CYTOCHROME C OXIDASE DEFICIENT1 gene leads to cytochrome c oxidase depletion and reorchestrated respiratory metabolism in Arabidopsis. (United States)

    Dahan, Jennifer; Tcherkez, Guillaume; Macherel, David; Benamar, Abdelilah; Belcram, Katia; Quadrado, Martine; Arnal, Nadège; Mireau, Hakim


    Cytochrome c oxidase is the last respiratory complex of the electron transfer chain in mitochondria and is responsible for transferring electrons to oxygen, the final acceptor, in the classical respiratory pathway. The essentiality of this step makes it that depletion in complex IV leads to lethality, thereby impeding studies on complex IV assembly and respiration plasticity in plants. Here, we characterized Arabidopsis (Arabidopsis thaliana) embryo-lethal mutant lines impaired in the expression of the CYTOCHROME C OXIDASE DEFICIENT1 (COD1) gene, which encodes a mitochondria-localized PentatricoPeptide Repeat protein. Although unable to germinate under usual conditions, cod1 homozygous embryos could be rescued from immature seeds and developed in vitro into slow-growing bush-like plantlets devoid of a root system. cod1 mutants were defective in C-to-U editing events in cytochrome oxidase subunit2 and NADH dehydrogenase subunit4 transcripts, encoding subunits of respiratory complex IV and I, respectively, and consequently lacked cytochrome c oxidase activity. We further show that respiratory oxygen consumption by cod1 plantlets is exclusively associated with alternative oxidase activity and that alternative NADH dehydrogenases are also up-regulated in these plants. The metabolomics pattern of cod1 mutants was also deeply altered, suggesting that alternative metabolic pathways compensated for the probable resulting restriction in NADH oxidation. Being the first complex IV-deficient mutants described in higher plants, cod1 lines should be instrumental to future studies on respiration homeostasis.

  8. Chromate reduction by rabbit liver aldehyde oxidase

    Energy Technology Data Exchange (ETDEWEB)

    Banks, R.B.; Cooke, R.T. Jr.


    Chromate was reduced during the oxidation of 1-methylnicotinamide chlorine by partially purified rabbit liver aldehyde oxidase. In addition to l-methylnicotinamide, several other electron donor substrates for aldehyde oxidase were able to support the enzymatic chromate reduction. The reduction required the presence of both enzyme and the electron donor substrate. The rate of the chromate reduction was retarded by inhibitors or aldehyde oxidase but was not affected by substrates or inhibitors of xanthine oxidase. These results are consistent with the involvement of aldehyde oxidase in the reduction of chromate by rabbit liver cytosolic enzyme preparations.

  9. Multiple amine oxidases in cucumber seedlings. (United States)

    Percival, F W; Purves, W K


    Cell-free extracts of cucumber (Cucumis sativus L. cv. National Pickling) seedlings were found to have amine oxidase activity when assayed with tryptamine as a substrate. Studies of the effect of lowered pH on the extract indicated that this activity was heterogeneous, and three amine oxidases could be separated by ion exchange chromatography. The partially purified enzymes were tested for their activities with several substrates and for their sensitivities to various amine oxidase inhibitors. One of the enzymes may be a monoamine oxidase, although it is inhibited by some diamine oxidase inhibitors. The other two enzymes have properties more characteristic of the diamine oxidases. The possible relationship of the amine oxidases to indoleacetic acid biosynthesis in cucumber seedlings is discussed.

  10. A Role for Reactive Oxygen Species Produced by NADPH Oxidases in the Embryo and Aleurone Cells in Barley Seed Germination.

    Directory of Open Access Journals (Sweden)

    Yushi Ishibashi

    Full Text Available Reactive oxygen species (ROS promote the germination of several seeds, and antioxidants suppress it. However, questions remain regarding the role and production mechanism of ROS in seed germination. Here, we focused on NADPH oxidases, which produce ROS. After imbibition, NADPH oxidase mRNAs were expressed in the embryo and in aleurone cells of barley seed; these expression sites were consistent with the sites of ROS production in the seed after imbibition. To clarify the role of NADPH oxidases in barley seed germination, we examined gibberellic acid (GA / abscisic acid (ABA metabolism and signaling in barley seeds treated with diphenylene iodonium chloride (DPI, an NADPH oxidase inhibitor. DPI significantly suppressed germination, and suppressed GA biosynthesis and ABA catabolism in embryos. GA, but not ABA, induced NADPH oxidase activity in aleurone cells. Additionally, DPI suppressed the early induction of α-amylase by GA in aleurone cells. These results suggest that ROS produced by NADPH oxidases promote GA biosynthesis in embryos, that GA induces and activates NADPH oxidases in aleurone cells, and that ROS produced by NADPH oxidases induce α-amylase in aleurone cells. We conclude that the ROS generated by NADPH oxidases regulate barley seed germination through GA / ABA metabolism and signaling in embryo and aleurone cells.

  11. A Role for Reactive Oxygen Species Produced by NADPH Oxidases in the Embryo and Aleurone Cells in Barley Seed Germination. (United States)

    Ishibashi, Yushi; Kasa, Shinsuke; Sakamoto, Masatsugu; Aoki, Nozomi; Kai, Kyohei; Yuasa, Takashi; Hanada, Atsushi; Yamaguchi, Shinjiro; Iwaya-Inoue, Mari


    Reactive oxygen species (ROS) promote the germination of several seeds, and antioxidants suppress it. However, questions remain regarding the role and production mechanism of ROS in seed germination. Here, we focused on NADPH oxidases, which produce ROS. After imbibition, NADPH oxidase mRNAs were expressed in the embryo and in aleurone cells of barley seed; these expression sites were consistent with the sites of ROS production in the seed after imbibition. To clarify the role of NADPH oxidases in barley seed germination, we examined gibberellic acid (GA) / abscisic acid (ABA) metabolism and signaling in barley seeds treated with diphenylene iodonium chloride (DPI), an NADPH oxidase inhibitor. DPI significantly suppressed germination, and suppressed GA biosynthesis and ABA catabolism in embryos. GA, but not ABA, induced NADPH oxidase activity in aleurone cells. Additionally, DPI suppressed the early induction of α-amylase by GA in aleurone cells. These results suggest that ROS produced by NADPH oxidases promote GA biosynthesis in embryos, that GA induces and activates NADPH oxidases in aleurone cells, and that ROS produced by NADPH oxidases induce α-amylase in aleurone cells. We conclude that the ROS generated by NADPH oxidases regulate barley seed germination through GA / ABA metabolism and signaling in embryo and aleurone cells.

  12. Processing optimization of probiotic yogurt containing glucose oxidase using response surface methodology. (United States)

    Cruz, A G; Faria, J A F; Walter, E H M; Andrade, R R; Cavalcanti, R N; Oliveira, C A F; Granato, D


    Exposure to oxygen may induce a lack of functionality of probiotic dairy foods because the anaerobic metabolism of probiotic bacteria compromises during storage the maintenance of their viability to provide benefits to consumer health. Glucose oxidase can constitute a potential alternative to increase the survival of probiotic bacteria in yogurt because it consumes the oxygen permeating to the inside of the pot during storage, thus making it possible to avoid the use of chemical additives. This research aimed to optimize the processing of probiotic yogurt supplemented with glucose oxidase using response surface methodology and to determine the levels of glucose and glucose oxidase that minimize the concentration of dissolved oxygen and maximize the Bifidobacterium longum count by the desirability function. Response surface methodology mathematical models adequately described the process, with adjusted determination coefficients of 83% for the oxygen and 94% for the B. longum. Linear and quadratic effects of the glucose oxidase were reported for the oxygen model, whereas for the B. longum count model an influence of the glucose oxidase at the linear level was observed followed by the quadratic influence of glucose and quadratic effect of glucose oxidase. The desirability function indicated that 62.32 ppm of glucose oxidase and 4.35 ppm of glucose was the best combination of these components for optimization of probiotic yogurt processing. An additional validation experiment was performed and results showed acceptable error between the predicted and experimental results.

  13. A Reflection on Teacher Questioning Types

    Directory of Open Access Journals (Sweden)

    Kasih Elisabeth Roostini


    Full Text Available Abstract This small-scale research is expected to serve as a reflective means for teachers so that they can explore their questioning types in their own classroom. It analyzed questioning types used by three teachers of general English classes. The questioning types were classified based on three dimensions—purpose, form, and function. The purpose-based questioning types, based on Long and Sato’s findings (1983, were classified into two: referential and display. The form-based questioning types, based on the classification of question types by Celce-Murcia and Larsen-Freeman (1999 and Biber et al. (1999, were classified into four major types: yes/no-questions, wh-questions, tag questions, and alternative questions. The function-based questioning types, based on Long and Sato’s findings (1983, were classified into three sub-types: comprehension checks, confirmation checks, and clarification requests.           The data, obtained from three recorded class meetings, were transcribed and analyzed to see what questioning types each teacher employed and to obtain the number and percentage distribution of questioning types each teacher used. The results showed that referential questions were more frequently used than display questions at higher levels. There were a substantial number of incomplete questions, a form-based questioning type that did not belong to the classification of form-based questioning types employed in this study. The predominant use of incomplete questions in a communicative classroom should be reviewed as this questioning type required accuracy, rather than promoting language practice. The third type, the function-based questioning types, did not occur frequently at all the three stages. This study suggests that teachers use incomplete questions less frequently and try to use other form-based questioning types. It also encourages teachers to reflect on their own teaching and pursue their professional development.

  14. Lysyl oxidase in cancer research. (United States)

    Perryman, Lara; Erler, Janine T


    Metastasis is the main reason for cancer-associated deaths and therapies are desperately needed to target the progression of cancer. Lysyl oxidase (LOX) plays a pivotal role in cancer progression, including metastasis, and is therefore is an attractive therapeutic target. In this review we will breakdown the process of cancer progression and the various roles that LOX plays has in the advancement of cancer. We will highlight why LOX is an exciting therapeutic target for the future.

  15. Characteristics of redox-linked proton ejection in cytochrome c oxidase reconstituted in phospholipid vesicles. New observations support mechanisms different from proton pumping. (United States)

    Papa, S; Lorusso, M; Capitanio, N; De Nitto, E


    Experimental observations reveal a number of characteristics of the redox-linked proton ejection from cytochrome c oxidase vesicles, which apparently cannot be explained by a proton pumping activity of the oxidase. These observations seem, on the other hand, to provide useful elements for alternative explanation(s) of the proton ejection. It is proposed here that the process is scalar and not vectorial and can derive from redox-linked rupture of protonated salt-bridges in the oxidase-lipid complex.

  16. HIF-1α activation by intermittent hypoxia requires NADPH oxidase stimulation by xanthine oxidase. (United States)

    Nanduri, Jayasri; Vaddi, Damodara Reddy; Khan, Shakil A; Wang, Ning; Makarenko, Vladislav; Semenza, Gregg L; Prabhakar, Nanduri R


    Hypoxia-inducible factor 1 (HIF-1) mediates many of the systemic and cellular responses to intermittent hypoxia (IH), which is an experimental model that simulates O2 saturation profiles occurring with recurrent apnea. IH-evoked HIF-1α synthesis and stability are due to increased reactive oxygen species (ROS) generated by NADPH oxidases, especially Nox2. However, the mechanisms by which IH activates Nox2 are not known. We recently reported that IH activates xanthine oxidase (XO) and the resulting increase in ROS elevates intracellular calcium levels. Since Nox2 activation requires increased intracellular calcium levels, we hypothesized XO-mediated calcium signaling contributes to Nox activation by IH. We tested this possibility in rat pheochromocytoma PC12 cells subjected to IH consisting alternating cycles of hypoxia (1.5% O2 for 30 sec) and normoxia (21% O2 for 5 min). Kinetic analysis revealed that IH-induced XO preceded Nox activation. Inhibition of XO activity either by allopurinol or by siRNA prevented IH-induced Nox activation, translocation of the cytosolic subunits p47phox and p67phox to the plasma membrane and their interaction with gp91phox. ROS generated by XO also contribute to IH-evoked Nox activation via calcium-dependent protein kinase C stimulation. More importantly, silencing XO blocked IH-induced upregulation of HIF-1α demonstrating that HIF-1α activation by IH requires Nox2 activation by XO.

  17. Alternative respiration and fumaric acid production of Rhizopus oryzae. (United States)

    Gu, Shuai; Xu, Qing; Huang, He; Li, Shuang


    Under the conditions of fumaric acid fermentation, Rhizopus oryzae ME-F14 possessed at least two respiratory systems. The respiration of mycelia was partially inhibited by the cytochrome respiration inhibitor antimycin A or the alternative respiration inhibitor salicylhydroxamic acid and was completely inhibited in the presence of both antimycin A and salicylhydroxamic acid. During fumaric acid fermentation process, the activity of alternative respiration had a great correlation with fumaric acid productivity; both of them reached peak at the same time. The alternative oxidase gene, which encoded the mitochondrial alternative oxidase responsible for alternative respiration in R. oryzae ME-F14, was cloned and characterized in Escherichia coli. The activity of alternative respiration, the alternative oxidase gene transcription level, as well as the fumaric acid titer were measured under different carbon sources and different carbon-nitrogen ratios. The activity of alternative respiration was found to be comparable to the transcription level of the alternative oxidase gene and the fumaric acid titer. These results indicated that the activity of the alternative oxidase was regulated at the transcription stage under the conditions tested for R. oryzae ME-F14.

  18. A Role for Reactive Oxygen Species Produced by NADPH Oxidases in the Embryo and Aleurone Cells in Barley Seed Germination


    Yushi Ishibashi; Shinsuke Kasa; Masatsugu Sakamoto; Nozomi Aoki; Kyohei Kai; Takashi Yuasa; Atsushi Hanada; Shinjiro Yamaguchi; Mari Iwaya-Inoue


    Reactive oxygen species (ROS) promote the germination of several seeds, and antioxidants suppress it. However, questions remain regarding the role and production mechanism of ROS in seed germination. Here, we focused on NADPH oxidases, which produce ROS. After imbibition, NADPH oxidase mRNAs were expressed in the embryo and in aleurone cells of barley seed; these expression sites were consistent with the sites of ROS production in the seed after imbibition. To clarify the role of NADPH oxidas...


    Energy Technology Data Exchange (ETDEWEB)



    PET is uniquely capable of providing information on biochemical transformations in the living human body. Although most of the studies of monoamine oxidase (MAO) have focused on measurements in the brain, the role of peripheral MAO as a phase 1 enzyme for the metabolism of drugs and xenobiotics is gaining attention (Strolin Benedetti and Tipton, 1998; Castagnoli et al., 1997.). MAO is well suited for this role because its concentration in organs such as kidneys, liver and digestive organs is high sometimes exceeding that in the brain. Knowledge of the distribution of the MAO subtypes within different organs and different cells is important in determining which substrates (and which drugs and xenobiotics) have access to which MAO subtypes. The highly variable subtype distribution with different species makes human studies even more important. In addition, the deleterious side effects of combining MAO inhibitors with other drugs and with foodstuffs makes it important to know the MAO inhibitory potency of different drugs both in the brain and in peripheral organs (Ulus et al., 2000). Clearly PET can play a role in answering these questions, in drug research and development and in discovering some of the factors which contribute to the highly variable MAO levels in different individuals.

  20. Revisiting Routine Questions (United States)

    Hughes, Rebecca; Monaghan, John; Shingadia, Eisha; Vaughan, Stephen


    What is a routine question? The focus of this paper is routine questions and time (in years) since a hitherto routine question was last attempted by the solver. The data comes from undergraduate students' work on solving two calculus questions. The data was selected for reporting purposes because it is well documented and because it threw up…

  1. Question analysis for Indonesian comparative question (United States)

    Saelan, A.; Purwarianti, A.; Widyantoro, D. H.


    Information seeking is one of human needs today. Comparing things using search engine surely take more times than search only one thing. In this paper, we analyzed comparative questions for comparative question answering system. Comparative question is a question that comparing two or more entities. We grouped comparative questions into 5 types: selection between mentioned entities, selection between unmentioned entities, selection between any entity, comparison, and yes or no question. Then we extracted 4 types of information from comparative questions: entity, aspect, comparison, and constraint. We built classifiers for classification task and information extraction task. Features used for classification task are bag of words, whether for information extraction, we used lexical, 2 previous and following words lexical, and previous label as features. We tried 2 scenarios: classification first and extraction first. For classification first, we used classification result as a feature for extraction. Otherwise, for extraction first, we used extraction result as features for classification. We found that the result would be better if we do extraction first before classification. For the extraction task, classification using SMO gave the best result (88.78%), while for classification, it is better to use naïve bayes (82.35%).

  2. 干旱胁迫下平邑甜茶叶片交替呼吸途径上调对光破坏的防御作用%Up-Regulation of the Mitochondrial Alternative Oxidase Pathway Enhances Photoprotection inMalus hupehensis Leaves under Drought Stress

    Institute of Scientific and Technical Information of China (English)

    徐秀玉; 孙山; 金立桥; 刘美君; 高辉远


    The role of mitochondrial alternative oxidase (AOX) pathway in photoprotection inMalus hupehen-sis leaves under drought stress was studied. The results indicated that drought increased the amount of leaf AOX protein and also enhanced the activity of AOX pathway. Under high light intensity, and after treated with salicylhydroxamic acid (SHAM) to inhibit the AOX pathway, the maximal photochemical efifciency of PSII (Fv/Fm), the J steps at the relative variable lfuorescence kinetics (VJ), the PSII excitation pressure (1–qP), and electron transport rate (ETR) was not affected in water-replete plants, while the ETR andFv/Fm signiifcantly de-creased, while the 1–qP andVJ signiifcantly increased in drought-stressed plants, indicating that under drought-stress condition, the inhibition of AOX pathway caused more severe photoinhibition. The results demonstrate that the AOX pathway plays an important role in the photoprotection inM. hupehensis leaves under drought stress.%以平邑甜茶为实验材料,研究了干旱胁迫下线粒体交替氧化酶呼吸途径(AOX途径)对平邑甜茶叶片的光破坏防御作用。结果表明:干旱胁迫下线粒体AOX的蛋白表达量及AOX途径的相对活性均显著增加;强光下用水杨基羟肟酸(SHAM)抑制正常供水和干旱胁迫处理的植株的AOX途径后,正常供水的平邑甜茶叶片的PSII最大光化学效率(Fv/Fm)、快速叶绿素荧光响应曲线中的J点相对可变荧光(VJ)、PSII反应中心关闭的比例(1–qP)和光合线性电子传递速率(ETR)几乎不受影响,而干旱胁迫下的平邑甜茶叶片的ETR和Fv/Fm显著下降,1–qP和VJ显著上升,这表明干旱胁迫下抑制AOX途径后,平邑甜茶叶片发生更严重的光抑制。上述结果表明:干旱胁迫下, AOX途径在平邑甜茶叶片的光破坏防御中起重要的作用。

  3. Multiple controls affect arsenite oxidase gene expression in Herminiimonas arsenicoxydans

    Directory of Open Access Journals (Sweden)

    Coppée Jean-Yves


    Full Text Available Abstract Background Both the speciation and toxicity of arsenic are affected by bacterial transformations, i.e. oxidation, reduction or methylation. These transformations have a major impact on environmental contamination and more particularly on arsenic contamination of drinking water. Herminiimonas arsenicoxydans has been isolated from an arsenic- contaminated environment and has developed various mechanisms for coping with arsenic, including the oxidation of As(III to As(V as a detoxification mechanism. Results In the present study, a differential transcriptome analysis was used to identify genes, including arsenite oxidase encoding genes, involved in the response of H. arsenicoxydans to As(III. To get insight into the molecular mechanisms of this enzyme activity, a Tn5 transposon mutagenesis was performed. Transposon insertions resulting in a lack of arsenite oxidase activity disrupted aoxR and aoxS genes, showing that the aox operon transcription is regulated by the AoxRS two-component system. Remarkably, transposon insertions were also identified in rpoN coding for the alternative N sigma factor (σ54 of RNA polymerase and in dnaJ coding for the Hsp70 co-chaperone. Western blotting with anti-AoxB antibodies and quantitative RT-PCR experiments allowed us to demonstrate that the rpoN and dnaJ gene products are involved in the control of arsenite oxidase gene expression. Finally, the transcriptional start site of the aoxAB operon was determined using rapid amplification of cDNA ends (RACE and a putative -12/-24 σ54-dependent promoter motif was identified upstream of aoxAB coding sequences. Conclusion These results reveal the existence of novel molecular regulatory processes governing arsenite oxidase expression in H. arsenicoxydans. These data are summarized in a model that functionally integrates arsenite oxidation in the adaptive response to As(III in this microorganism.

  4. The question of questions in Malaysian English


    Asniah Alias; Radina Mohamad Deli


    This paper examined the forms that interrogatives and tag questions can take when used by young Malaysian speakers of English language in oral communication. It offers a description of the features for both question forms as produced by the respondents compared to those of Singapore English (SE) and Standard British English (SBE). The influence of domains and the issue of mother tongue interference in relation to the subjects’ usage of such features will also be investigated. Data were obtain...

  5. The reaction of Pseudomonas aeurginosa cytochrome c oxidase with sodium metabisulphite. (United States)

    Parr, S R; Wilson, M T; Greenwood, C


    Spectrophotometric evidence is presented for the formation of a complex between metabisulphite and reduced Pseudomonas aeruginosa cytochrome c oxidase. The effects of metabisulphite on the recombination of CO with the reduced enzyme are discussed in terms of alternate binding sites for S(2)O(5) (2-) and CO.

  6. Vaginal dryness alternative treatments (United States)

    Alternative treatments for vaginal dryness ... Question: Is there a drug-free treatment for vaginal dryness? Answer: There are many causes of vaginal dryness . It may be caused by reduced estrogen level, infection, medicines, and ...

  7. Oxidase-based biocatalytic processes

    DEFF Research Database (Denmark)

    Ramesh, Hemalata; Woodley, John; Krühne, Ulrich

    the reaction species (substrate and product volatility for example) and the process (such as oxygen supply, ability to control pH) and are classified as reaction-related and process-related constraintsrespectively. Although the development of biocatalyst and process engineering tools offers a number...... ofsolutions to overcome the limitations, it is often complicated to identify the key limitation of the system that prevents economic scale-up. Hence, development of a systematic method for identifying the limitations during early-stage development of a biocatalytic process and potentially the order in which...... theyneed to be tackled would offer a valuable tool for process development.Biocatalytic oxidationsare potentially of great value because of theselective chemistry that they offer,resulting in higher yieldscompared to thoseachievable through chemical catalysis. Oxidases areparticularly...

  8. A Technique Socratic Questioning-Guided Discovery

    Directory of Open Access Journals (Sweden)

    M. Hakan Türkçapar


    Full Text Available “Socratic Method” is a way of teaching philosophical thinking and knowledge by asking questions which was used by antique period greek philosopher Socrates. Socrates was teaching knowledge to his followers by asking questions and the conversation between them was named “Socratic Dialogues”. In this meaning, no novel knowledge is taught to the individual but only what is formerly known is reminded and rediscovered. The form of socratic questioning which is used during the process of cognitive behavioral therapy is known as Guided Discovery. In this method it is aimed to make the client notice the piece of knowledge which he could notice but is not aware with a series of questions. Socratic method or guided discovery consists of several steps which are: Identifying the problem by listening to the client and making reflections, finding alternatives by examining and evaluating, reidentification by using the newly found information and questioning the old distorted belief and reaching to a conclusion and applying it. Question types used during these procedures are, questions for gaining information, questions revealing the meanings, questions revealing the beliefs, questions about behaviours during the similar past experiences, analyse questions and analytic synthesis questions. In order to make the patient feel understood it is important to be empathetic and summarising the problem during the interview. In this text, steps of Socratic Questioning-Guided Discovery will be reviewed with sample dialogues after each step

  9. Classroom Questioning Strategies

    Institute of Scientific and Technical Information of China (English)



      Interaction has been playing a more and more important role in language research since the early 1970s,when the communicative teaching method was widely applied in language teaching. Questioning is the most common classroom interaction. This thesis analyzed the influence on students' immediate oral production by applying different teacher questioning strategies including teacher's question types,teacher question modification and teacher feedback.

  10. Asking Questions in Academia

    DEFF Research Database (Denmark)

    Hansen, Annette Skovsted


    Motivation for the activity In academia the most important skill is to ask academically relevant and sound questions. This is not easy and students need to practice asking questions orally and in writing before they write research papers.......Motivation for the activity In academia the most important skill is to ask academically relevant and sound questions. This is not easy and students need to practice asking questions orally and in writing before they write research papers....

  11. Exam Question Exchange. (United States)

    Alexander, John J., Ed.


    Acceptable answers are provided for two chemistry questions. The first question is related to the prediction of the appearance of non-first-order proton nuclear magnetic resonance (NMR) spectra. The second question is related to extraterrestrial kinetic theory of gases. (JN)

  12. Interfacing with questions

    DEFF Research Database (Denmark)

    Soon, Winnie


    This article discusses an artistic project entitled If I wrote you a love letter would you write back (and thousands of other questions): a piece of software that utilizes Twitter web API to query questions, drawing unpredictable questions in real-time from the distributed database of Twitter...

  13. Improving Student Question Classification (United States)

    Heiner, Cecily; Zachary, Joseph L.


    Students in introductory programming classes often articulate their questions and information needs incompletely. Consequently, the automatic classification of student questions to provide automated tutorial responses is a challenging problem. This paper analyzes 411 questions from an introductory Java programming course by reducing the natural…

  14. Question-Asking and Question-Exploring (United States)

    Sands, Lorraine; Carr, Margaret; Lee, Wendy


    The Centre of Innovation Research at Greerton Early Childhood Centre was characterised as a dispositional milieu where working theories were explored through a narrative research methodology. As the research progressed, the teachers at Greerton strengthened the way we were listening to, and watching out for young children's questions to enable…

  15. Unproven (questionable) cancer therapies.


    Brigden, M.L.


    More than half of all cancer patients use some form of alternative treatment during the course of their illness. Alternative therapies are often started early in patients' illness, and their use is frequently not acknowledged to health care professionals. Some alternative therapies are harmful, and their promoters may be fraudulent. Persons who try alternative cancer therapies may not be poorly educated but may ultimately abandon conventional treatment. Recent attention has focused on aspects...

  16. Vanillyl-alcohol oxidase, a tasteful biocatalyst

    NARCIS (Netherlands)

    Heuvel, Robert H.H. van den; Fraaije, Marco W.; Mattevi, Andrea; Laane, Colja; Berkel, Willem J.H. van


    The covalent flavoenzyme vanillyl-alcohol oxidase (VAO) is a versatile biocatalyst. It converts a wide range of phenolic compounds by catalysing oxidation, deamination, demethylation, dehydrogenation and hydroxylation reactions. The production of natural vanillin, 4-hydroxybenzaldehyde, coniferyl al

  17. Galactose oxidase nanoaggregates: Preparation and characterization


    Mamta Sharma; Minakshi Sharma


    Galctose oxidase nanoaggregates have been prepared by chemical desolvation method involving the crosslinkng agent glutaraldehyde. These enzyme nanoagregates have been characterized by transmission electron microscopy(TEM), UV visible spectroscopy, Fourier transform infrared spectroscopy (FTIR). TEM reveals the globular spherical nanostructured form upto the range of 20nm. UV visible spectroscopy of galactose oxidase nanoaggregates shows maximum absorption peak at 237nm. FTIR spectra obtaine...

  18. Redox-Dependent Conformational Changes in Cytochrome c Oxidase Suggest a Gating Mechanism for Proton Uptake

    Energy Technology Data Exchange (ETDEWEB)

    Qin, Ling; Liu, Jian; Mills, Denise A.; Proshlyakov, Denis A.; Hiser, Carrie; Ferguson-Miller, Shelagh; (MSU)


    A role for conformational change in the coupling mechanism of cytochrome c oxidase is the subject of controversy. Relatively small conformational changes have been reported in comparisons of reduced and oxidized crystal structures of bovine oxidase but none in bacterial oxidases. Comparing the X-ray crystal structures of the reduced (at 2.15 {angstrom} resolution) and oxidized forms of cytochrome c oxidase from Rhodobacter sphaeroides, we observe a displacement of heme a3 involving both the porphyrin ring and the hydroxyl farnesyl tail, accompanied by protein movements in nearby regions, including the mid part of helix VIII of subunit I which harbors key residues of the K proton uptake path, K362 and T359. The conformational changes in the reduced form are reversible upon reoxidation. They result in an opening of the top of the K pathway and more ordered waters being resolved in that region, suggesting an access path for protons into the active site. In all high-resolution structures of oxidized R. sphaeroides cytochrome c oxidase, a water molecule is observed in the hydrophobic region above the top of the D path, strategically positioned to facilitate the connection of residue E286 of subunit I to the active site or to the proton pumping exit path. In the reduced and reduced plus cyanide structures, this water molecule disappears, implying disruption of proton conduction from the D path under conditions when the K path is open, thus providing a mechanism for alternating access to the active site.

  19. Fluorescence quenching study of quercetin interaction with bovine milk xanthine oxidase (United States)

    Rasoulzadeh, Farzaneh; Jabary, Hamideh Nadjarpour; Naseri, Abdolhossein; Rashidi, Mohammad-Reza


    Quercetin is a natural flavonoid with many important therapeutic properties. The interaction of this polyphenolic compound bovine milk xanthine oxidase as one of its major target proteins was studied using fluorescence quenching method for the first time. It was found that the fluorescence quenching of xanthine oxidase occurs through a static mechanism. The results revealed the presence of a single binding site on xanthine oxidase with the binding constant value equals to 1.153 × 10 4 l mol -1 at 310 K and pH 7.4. The thermodynamic parameters were also calculated at different temperatures. The enthalpy and entropy changes were found as -10.661 kJ mol -1 and +43.321 J mol -1 K -1 indicating that both hydrogen binding and hydrophobic are involved in the interaction of this polyphenolic natural compound with xanthine oxidase. The results may provide a ground for further studies with different flavonoids to find a safe alternative for allopurinol, the only xanthine oxidase inhibitor with clinical application.

  20. Questions and information systems

    CERN Document Server

    Lauer, Thomas W; Graesser, Arthur C


    The design and functioning of an information system improve to the extent that the system can handle the questions people ask. Surprisingly, however, researchers in the cognitive, computer, and information sciences have not thoroughly examined the multitude of relationships between information systems and questions -- both question asking and answering. The purpose of this book is to explicitly examine these relationships. Chapter contributors believe that questions play a central role in the analysis, design, and use of different kinds of natural or artificial information systems such as huma

  1. Asking the Right Questions. (United States)

    Galvin, Ruth Mehrtens


    Like good researchers, writers about research need to be able to tell whether scientific findings are valid. Six questions, to be asked of the researcher, can help the writer explain to others. A healthy skepticism is also important; there may be signals that more questions should be asked. (MSE)

  2. Epistemology: 5 Questions

    DEFF Research Database (Denmark)

    Epistemology: 5 Questions is a collection of short interviews based on 5 questions presented to some of the most influential and prominent scholars in epistemology. We hear their views on epistemology with particular emphasis on the intersection between mainstream and formal approaches to the field......; the aim, scope, the future direction of epistemology and how their work fits in these respects...

  3. Legal Philosophy - Five Questions

    DEFF Research Database (Denmark)

    This collection gathers together a host of the most eminent contemporary legal philosophers, who writes about their take on legal philosophy, its fundamental questions and potential.......This collection gathers together a host of the most eminent contemporary legal philosophers, who writes about their take on legal philosophy, its fundamental questions and potential....

  4. Let's Switch Questioning Around (United States)

    Tovani, Cris


    English teacher Cris Tovani knows from her experiences teaching elementary school that students are naturally curious. But, too often, students are so trained to be question answerers that by the time they reach high school, they no longer form questions of their own and instead focus on trying to figure out what answer the teacher wants. Tovani…

  5. What is a Question? (United States)

    Knuth, Kevin H.; Clancy, Daniel (Technical Monitor)


    A given question can be defined in terms of the set of statements or assertions that answer it. Application of the logic of inference to this set of assertions allows one to derive the logic of inquiry among questions. There are interesting symmetries between the logics of inference and inquiry; where probability describes the degree to which a premise implies an assertion, there exists an analogous quantity that describes the bearing or relevance that a question has on an outstanding issue. These have been extended to suggest that the logic of inquiry results in functional relationships analogous to, although more general than, those found in information theory. Employing lattice theory, I examine in greater detail the structure of the space of assertions and questions demonstrating that the symmetries between the logical relations in each of the spaces derive directly from the lattice structure. Furthermore, I show that while symmetries between the spaces exist, the two lattices are not isomorphic. The lattice of assertions is described by a Boolean lattice 2(sup N) whereas the lattice of real questions is shown to be a sublattice of the free distributive lattice FD(N) = 2(sup 2(sup N)). Thus there does not exist a one-to-one mapping of assertions to questions, there is no reflection symmetry between the two spaces, and questions in general do not possess unique complements. Last, with these lattice structures in mind, I discuss the relationship between probability, relevance and entropy.


    Institute of Scientific and Technical Information of China (English)



    Under more general form and more general conditions an affirmative answer to Reich's open question is given. The results presented also extend and improve some recent results of Reich, Shioji, Takahashi and Wittmann.

  7. multicopper oxidases important for human iron metabolism

    Directory of Open Access Journals (Sweden)

    Diana Wierzbicka


    Full Text Available Multi-copper oxidases are a group of proteins which demonstrate enzymatic activity and are capable of oxidizing their substrates with the concomitant reduction of dioxygen to two water molecules. For some multi-copper oxidases there has been demonstrated ferroxidase activity which is related to their specific structure characterized by the presence of copper centres and iron-binding sites. Three multi-copper oxidases have been included in this group: ceruloplasmin, hephaestin and zyklopen. Multi copper oxidases which are expressed in different tissues are capable of oxidizing a wide spectrum of substrates. Multi-copper oxidases are capable of oxidizing a wide spectrum of substrates. Ceruloplasmin exhibits antioxidant activity as well as being involved in many other biological processes. The observations of phenotypic effects of absence or low expression of multi-copper ferroxidase-coding genes suggest that the main role of these proteins is taking part in iron metabolism. The main role of ceruloplasmin in iron turnover is oxidizing Fe2+ into Fe3+, a process which is essential for iron binding to transferrin (the main iron-transporting protein, as well as to ferritin (the main iron-storage protein. The function of hephaestin as ferroxidase is essential for iron binding to apotransferrin in the lamina propria of the intestinal mucosa, a process that is important for further transport of iron to the liver by the portal vein. Available data indicate that zyklopen is responsible for the placental iron transport. The presence of three multi-copper oxidases with ferroxidase activity emphasizes the significance of oxidation for iron metabolism. The distribution of multi-copper ferroxidases in many tissues ensures the proper iron turnover in the body as well as preventing toxic effects related to the presence of Fe2+ ions. These ions contribute to generation of free radicals, including the highly reactive hydroxyl radical, through the Fenton and Haber

  8. Molecular evolution of the polyamine oxidase gene family in Metazoa

    Directory of Open Access Journals (Sweden)

    Polticelli Fabio


    Full Text Available Abstract Background Polyamine oxidase enzymes catalyze the oxidation of polyamines and acetylpolyamines. Since polyamines are basic regulators of cell growth and proliferation, their homeostasis is crucial for cell life. Members of the polyamine oxidase gene family have been identified in a wide variety of animals, including vertebrates, arthropodes, nematodes, placozoa, as well as in plants and fungi. Polyamine oxidases (PAOs from yeast can oxidize spermine, N1-acetylspermine, and N1-acetylspermidine, however, in vertebrates two different enzymes, namely spermine oxidase (SMO and acetylpolyamine oxidase (APAO, specifically catalyze the oxidation of spermine, and N1-acetylspermine/N1-acetylspermidine, respectively. Little is known about the molecular evolutionary history of these enzymes. However, since the yeast PAO is able to catalyze the oxidation of both acetylated and non acetylated polyamines, and in vertebrates these functions are addressed by two specialized polyamine oxidase subfamilies (APAO and SMO, it can be hypothesized an ancestral reference for the former enzyme from which the latter would have been derived. Results We analysed 36 SMO, 26 APAO, and 14 PAO homologue protein sequences from 54 taxa including various vertebrates and invertebrates. The analysis of the full-length sequences and the principal domains of vertebrate and invertebrate PAOs yielded consensus primary protein sequences for vertebrate SMOs and APAOs, and invertebrate PAOs. This analysis, coupled to molecular modeling techniques, also unveiled sequence regions that confer specific structural and functional properties, including substrate specificity, by the different PAO subfamilies. Molecular phylogenetic trees revealed a basal position of all the invertebrates PAO enzymes relative to vertebrate SMOs and APAOs. PAOs from insects constitute a monophyletic clade. Two PAO variants sampled in the amphioxus are basal to the dichotomy between two well supported

  9. Low platelet monoamine oxidase activity in pathological gambling

    Energy Technology Data Exchange (ETDEWEB)

    Carrasco, J.L. [Department of Psychiatry, Centro de Salud Mental, Parla Madrid (Spain); Saiz-Ruiz, J. [Department of Psychiatry and Haematology, Hospital Ramon y Cajal, Madrid (Spain); Hollander, E. [Department of Psychiatry, Mount Sinai School of Medicine, Queens Hospital Center, New York (United States); Cesar, J. [Department of Haematology, Hospital Ramon y Cajal, Madrid (Spain); Lopez-Ibor, J.J. Jr. [Department of Psychiatry, Hospital San Carlos, Complutense University, Madrid (Spain)


    Decreased platelet monoamine oxidase (MAO) activity has been reported in association with sensation-seeking personality type and in some mental disorders associated with a lack of impulse control. Pathological gambling itself has been related with both sensation-seeking and reduced impulse control. Platelet MAO activity was investigated in 15 DSM-III-R pathological gamblers from our outpatient clinic. Gamblers had a significantly lower platelet MAO activity than a group of 25 healthy controls. The range of MAO levels in gamblers was also significantly shorter than in controls. In controls, platelet MAO levels showed the previously described negative correlations with sensation-seeking scores but not in gamblers. The findings are consistent with previous studies showing an association of low platelet MAO activity with impulse control disorders and raise some interesting therapeutic alternatives for pathological gambling. (au) (40 refs.).

  10. Alternative energies; Energies alternatives

    Energy Technology Data Exchange (ETDEWEB)

    Bonal, J.; Rossetti, P


    The earth took millions years to made the petroleum, the gas the coal and the uranium. Only a few centuries will be needed to exhaust these fossil fuels and some years to reach expensive prices. Will the wold continue on this way of energy compulsive consumption? The renewable energies and some citizen attitudes are sufficient to break this spiral. This book proposes to discuss these alternative energies. It shows that this attitude must be supported by the government. It takes stock on the more recent information concerning the renewable energies. it develops three main points: the electricity storage, the housing and the transports. (A.L.B.)

  11. Engineering pyranose 2-oxidase for modified oxygen reactivity.

    Directory of Open Access Journals (Sweden)

    Dagmar Brugger

    Full Text Available Pyranose 2-oxidase (POx, a member of the GMC family of flavoproteins, catalyzes the regioselective oxidation of aldopyranoses at position C2 to the corresponding 2-ketoaldoses. During the first half-reaction, FAD is reduced to FADH2 and reoxidized in the second half-reaction by reducing molecular oxygen to H2O2. Alternative electron acceptors including quinones, radicals or chelated metal ions show significant and in some cases even higher activity. While oxygen as cheap and abundantly available electron acceptor is favored for many processes, reduced oxygen reactivity is desirable for some applications such as in biosensors/biofuel cells because of reduced oxidative damages to the biocatalyst from concomitant H2O2 production as well as reduced electron "leakage" to oxygen. The reactivity of flavoproteins with oxygen is of considerable scientific interest, and the determinants of oxygen activation and reactivity are the subject of numerous studies. We applied site-saturation mutagenesis on a set of eleven amino acids around the active site based on the crystal structure of the enzyme. Using microtiter plate screening assays with peroxidase/2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid and 2,6-dichlorophenolindophenol, variants of POx with decreased oxidase activity and maintained dehydrogenase activity were identified. Variants T166R, Q448H, L545C, L547R and N593C were characterized with respect to their apparent steady-state constants with oxygen and the alternative electron acceptors DCPIP, 1,4-benzoquinone and ferricenium ion, and the effect of the mutations was rationalized based on structural properties.

  12. Asking the Right Questions

    DEFF Research Database (Denmark)

    Richter, Line

    are part of everyday life, children are often the most vulnerable. The project was carried out to shed light on mainly two types of diseases - malaria and diarrheal diseases - that strike children. In practice the academic backgrounds of the researchers played a role in the methodological approach...... to the field. By emphasizing the logos in methodology this paper wishes to underscore that where anthropology sets itself apart from public health is, among other, in the way anthropologists think about method and how this affects fieldwork practices as well as analyses. By tracing two concepts, hygiene......, is the ability to move beyond even the best hidden assumptions and question our own questions, thereby enabling us to ask the right questions....

  13. 21 CFR 866.2420 - Oxidase screening test for gonorrhea. (United States)


    ... Section 866.2420 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Microbiology Devices § 866.2420 Oxidase... ingredient that will react with cytochrome oxidase. When cytochrome oxidase is present, the swab turns a dark...

  14. Bilirubin Oxidase Activity of Bacillus subtilis CotA


    Sakasegawa, S; Ishikawa, H.; Imamura, S.; Sakuraba, H.; Goda, S.; Ohshima, T.


    The spore coat protein CotA from Bacillus subtilis was previously identified as a laccase. We have now found that CotA also shows strong bilirubin oxidase activity and markedly higher affinity for bilirubin than conventional bilirubin oxidase. This is the first characterization of bilirubin oxidase activity in a bacterial protein.

  15. Kinetic mechanism of putrescine oxidase from Rhodococcus erythropolis

    NARCIS (Netherlands)

    Kopacz, Malgorzata; Heuts, Dominic P. H. M.; Fraaije, Marco W.


    Putrescine oxidase from Rhodococcus erythropolis (PuO) is a flavin-containing amine oxidase from the monoamine oxidase family that performs oxidative deamination of aliphatic diamines. In this study we report pre-steady-state kinetic analyses of the enzyme with the use of single-and double-mixing st


    NARCIS (Netherlands)

    Elferink, Marieke G.L.; Bosmal, Tjibbe; Lolkema, Juke S.; Gleiszner, Michael; Driessen, Arnold J.M.; Konings, Wil N.


    The effect of the lipid environment on the thermostability of three respiratory terminal oxidases was determined. Cytochrome-e oxidase from beef heart and Bacillus stearothermophilus were used as representative proteins from mesophilic and thermophilic origin, respectively. Quinol oxidase from the a

  17. Kinetic mechanism of putrescine oxidase from Rhodococcus erythropolis

    NARCIS (Netherlands)

    Kopacz, Malgorzata; Heuts, Dominic P. H. M.; Fraaije, Marco W.


    Putrescine oxidase from Rhodococcus erythropolis (PuO) is a flavin-containing amine oxidase from the monoamine oxidase family that performs oxidative deamination of aliphatic diamines. In this study we report pre-steady-state kinetic analyses of the enzyme with the use of single-and double-mixing

  18. The Question Concerning Thinking

    DEFF Research Database (Denmark)

    Riis, Søren


    Martin Heidegger's thought-provoking essay "The Question Concerning Technology" (1977a) placed technology at the heart of philosophy. Heidegger tried to show that the essence of technology provokes humans to think about the world in a very dangerous way. Yet if we follow Heidegger's analysis......, by doing so, we will in the end realize two important things. First, that Heidegger's declaration of the end of philosophy in fact also means the end of anything we can meaningfully call thinking. Second, that Heidegger's own thinking is completely different from his own ideal of thinking. Our question...

  19. The social question revisited

    DEFF Research Database (Denmark)

    Kenneth, Petersson; Olsson, Ulf; Krejsler, John B.


    The focus of this chapter is the re-installation of the social question as a historical practice. The purpose is to investigate how historic figures return and are applied in contemporary political discourses, more precisely in the context of education, education policy and teacher education...... the diversity of relevant populations “without obstacles related to their social and economic background”. In the 19th century the social question was raised in a context of industrialization of societies. It dealt with suggestions about disintegration of predominant social structures and the management...

  20. Alternative additives; Alternative additiver

    Energy Technology Data Exchange (ETDEWEB)


    In this project a number of industrial and agricultural waste products have been characterised and evaluated in terms of alkali-getter performance. The intended use is for biomass-fired power stations aiming at reducing corrosion or slagging related problems. The following products have been obtained, characterised and evaluated: 1) Brewery draff 2) Danish de-gassed manure 3) Paper sludge 4) Moulding sand 5) Spent bleaching earth 6) Anorthosite 7) Sand 8) Clay-sludge. Most of the above alternative additive candidates are deemed unsuitable due to insufficient chemical effect and/or expensive requirements for pre-treatment (such as drying and transportation). 3 products were selected for full-scale testing: de-gassed manure, spent bleaching earth and clay slugde. The full scale tests were undertaken at the biomass-fired power stations in Koege, Slagelse and Ensted. Spent bleaching earth (SBE) and clay sludge were the only tested additive candidates that had a proven ability to react with KCl, to thereby reduce Cl-concentrations in deposits, and reduce the deposit flux to superheater tubes. Their performance was shown to nearly as good as commercial additives. De-gassed manure, however, did not evaluate positively due to inhibiting effects of Ca in the manure. Furthermore, de-gassed manure has a high concentration of heavy metals, which imposes a financial burden with regard to proper disposal of the ash by-products. Clay-sludge is a wet clay slurring, and drying and transportation of this product entails substantial costs. Spent bleaching does not require much pre-treatment and is therefore the most promising alternative additive. On the other hand, bleaching earth contains residual plant oil which means that a range of legislation relating to waste combustion comes into play. Not least a waste combustion fee of 330 DKK/tonne. For all alternative (and commercial) additives disposal costs of the increase ash by-products represents a significant cost. This is

  1. Occurrence and Biocatalytic Potential of Carbohydrate Oxidases.

    NARCIS (Netherlands)

    Hellemond, van E.W.; Leferink, N.G.H.; Heuts, D.P.H.M.; Fraaije, M.W.; Berkel, van W.J.H.


    Carbohydrate oxidases are found in all kingdoms of life but are mostly found in fungi. Their natural role is not always clear. Usage of molecular oxygen as electron acceptor is not a logical choice when the enzyme is part of a catabolic pathway. This chapter provides an overview of the occurrence

  2. A colorimetric assay for cytokinin oxidase. (United States)

    Libreros-Minotta, C A; Tipton, P A


    A simple and rapid colorimetric assay for cytokinin oxidase is described. The assay is based on the formation of a Schiff base between the enzymatic reaction product 3-methyl-2-butenal and p-aminophenol. The assay is effective in the submicromolar concentration range and can be used in crude plant extracts as well as in more highly purified preparations.

  3. Spectrophotometric Assay of Immobilized Glucose Oxidase

    Directory of Open Access Journals (Sweden)

    Nojan Noorbehesht


    Full Text Available Enzyme results in change the substrate of product. Each enzyme may act on specific substrates, resulting in product or different products. The enzyme glucose oxidase (GOX is a bio catalyst. It accelerates the process of transforming glucose into hydrogen peroxide (H2O2 . These enzymes are used in the chemical industry, food industry, cosmetics and kits for diagnosis of glucose. There are many researches about immobilizations of Glucose Oxide to increase specifications such as repeated use, recovery, stability, shelf life and other features In this work, glucose oxidase enzyme using covalent bonding is placed on the carrier of carbon nanotubes. In this study, multi-walled carbon nanotubes have been used as adsorbents. Also, carbon nanotubes have been functionalized by sulfuric acid and nitric acid with a high concentration. Glucose oxidase is a biological biocatalyst enzyme. It accelerates changing glucose to H2O2. This enzyme is used in the chemical industry, food industry, cosmetics and glucose diagnostic kits. For example, as a result of ongoing research working focuses on the development of glucose biosensors, GOX in practice as standard enzyme has been revealed for immobilization of oxidative enzyme.GOX correct fixation on the MWNTs carrier is a way to reuse enzyme and miniature of biosensor devices and structures. In this study, a spectrophotometer was used to determine the absorbance of the enzyme glucose oxidase (GOX to review its activities after stabilizing the carbon nanotubes.

  4. Lysyl oxidase mediates hypoxic control of metastasis

    DEFF Research Database (Denmark)

    Erler, Janine Terra; Giaccia, Amato J


    Hypoxic cancer cells pose a great challenge to the oncologist because they are especially aggressive, metastatic, and resistant to therapy. Recently, we showed that elevation of the extracellular matrix protein lysyl oxidase (LOX) correlates with metastatic disease and is essential for hypoxia...

  5. A topic clustering approach to finding similar questions from large question and answer archives.

    Directory of Open Access Journals (Sweden)

    Wei-Nan Zhang

    Full Text Available With the blooming of Web 2.0, Community Question Answering (CQA services such as Yahoo! Answers (, WikiAnswer (, and Baidu Zhidao (, etc., have emerged as alternatives for knowledge and information acquisition. Over time, a large number of question and answer (Q&A pairs with high quality devoted by human intelligence have been accumulated as a comprehensive knowledge base. Unlike the search engines, which return long lists of results, searching in the CQA services can obtain the correct answers to the question queries by automatically finding similar questions that have already been answered by other users. Hence, it greatly improves the efficiency of the online information retrieval. However, given a question query, finding the similar and well-answered questions is a non-trivial task. The main challenge is the word mismatch between question query (query and candidate question for retrieval (question. To investigate this problem, in this study, we capture the word semantic similarity between query and question by introducing the topic modeling approach. We then propose an unsupervised machine-learning approach to finding similar questions on CQA Q&A archives. The experimental results show that our proposed approach significantly outperforms the state-of-the-art methods.

  6. Questioning Danish Cartoon Images

    DEFF Research Database (Denmark)

    Bojsen, Heidi


    Danes today when it is considered demeaning and racist in most other countries. The conclusion does emphatically not plead in favour of law enforced limitations of the freedom of expression, but does question the prevalent "freedom of ignorance" concerning black identities which means...

  7. Questions English Teachers Ask. (United States)

    Shuman, R. Baird

    This volume is based on the responses of 374 English teachers at the secondary and college levels to a letter asking them to describe the questions that most perplex them professionally. Answers are provided by 88 leaders in English education, including James R. Squire, Walter H. MacGinitie, R. Baird Shuman, Sheila Schwartz, and Ken Macrorie. The…

  8. Game Theory: 5 Questions

    DEFF Research Database (Denmark)

    Hendricks, Vincent F.

    Game Theory is a collection of short interviews based on 5 questions presented to some of the most influential and prominent scholars in game theory. We hear their views on game theory, its aim, scope, use, the future direction of game theory and how their work fits in these respects....

  9. Question: Who Can Vote? (United States)

    Rodeheaver, Misty D.; Haas, Mary E.


    This year's rollercoaster primary elections and the pending national election, with an anticipated record voter turnout, provide the perfect backdrop for an examination of the questions: (1) Who can vote?; and (2) Who will vote? Historically, the American government refused voting rights to various groups based on race, gender, age, and even…

  10. Future Research Questions

    NARCIS (Netherlands)

    Walpoth, B.; Daanen, H.A.M.


    Immersion hypothermia in humans is described in about 500 scientific papers during a Pubmed search in medical literature with keywords ‘Immersion’, ‘Hypothermia’ and ‘Human’ as of 2014. Many questions still remain, the most important of which are described in this chapter.

  11. Game Theory: 5 Questions

    DEFF Research Database (Denmark)

    Hendricks, Vincent F.

    Game Theory is a collection of short interviews based on 5 questions presented to some of the most influential and prominent scholars in game theory. We hear their views on game theory, its aim, scope, use, the future direction of game theory and how their work fits in these respects....

  12. Structure-function characterization reveals new catalytic diversity in the galactose oxidase and glyoxal oxidase family. (United States)

    Yin, DeLu Tyler; Urresti, Saioa; Lafond, Mickael; Johnston, Esther M; Derikvand, Fatemeh; Ciano, Luisa; Berrin, Jean-Guy; Henrissat, Bernard; Walton, Paul H; Davies, Gideon J; Brumer, Harry


    Alcohol oxidases, including carbohydrate oxidases, have a long history of research that has generated fundamental biological understanding and biotechnological applications. Despite a long history of study, the galactose 6-oxidase/glyoxal oxidase family of mononuclear copper-radical oxidases, Auxiliary Activity Family 5 (AA5), is currently represented by only very few characterized members. Here we report the recombinant production and detailed structure-function analyses of two homologues from the phytopathogenic fungi Colletotrichum graminicola and C. gloeosporioides, CgrAlcOx and CglAlcOx, respectively, to explore the wider biocatalytic potential in AA5. EPR spectroscopy and crystallographic analysis confirm a common active-site structure vis-à-vis the archetypal galactose 6-oxidase from Fusarium graminearum. Strikingly, however, CgrAlcOx and CglAlcOx are essentially incapable of oxidizing galactose and galactosides, but instead efficiently catalyse the oxidation of diverse aliphatic alcohols. The results highlight the significant potential of prospecting the evolutionary diversity of AA5 to reveal novel enzyme specificities, thereby informing both biology and applications.

  13. Status and Advances of Researches on GA 20-oxidases

    Institute of Scientific and Technical Information of China (English)

    Li Wei; Chen Xiaoyang; Li Hui; Guo Hai


    GA 20-oxidase, the most important limiting enzyme, can catalyze a series of oxidization of GA biosynthesis pathwayfrom GA12 to GA9 and from GA53 to GA20 in the higher plants. This paper reviews the studies on the characters of GA 20-oxidase,the gene and the protein of GA 20-oxidase and the regulation of GA 20-oxidase gene expression in recent years. At the same time,the prospects for the gene transformation of GA 20-oxidase in agriculture, forestry and horticulture are also discussed.


    Energy Technology Data Exchange (ETDEWEB)

    Swanson, Maurice; Packer, Lester


    Purified and reconstituted cytochrome {und c} oxidase and mitochondria were crosslinked with biimidates in the presence and absence of cytochrome {und c}. These experiments indicate that oxidase subunit interactions are required for activity and that cytochrome {und c} mobility may be required for electron transport activity. Biimidate treatment of purified and reconstituted oxidase crosslinks all of the oxidase protomers except subunit I when {ge} 20% of the free amines are modified and inhibits steady state oxidase activity. Transient kinetics of ferrocytochrome {und c} oxidation and ferricytochrome {und a} reduction indicates inhibition of electron transfer from heme {und a} to heme {und a}{sub 3}. Crosslinking oxidase molecules to form large aggregates displaying rotational correlation times {ge} 1 ms does not affect oxidase activity. Crosslinking of mitochondria covalently binds the bc{sub 1} and {und aa}{sub 3} complexes to cytochrome {und c}, and inhibits steady-state oxidase activity considerably more than in the case of the purified oxidase. Addition of cytochrome {und c} to the purified oxidase or to {und c}-depleted mitoplasts increases inhibition slightly. Cytochrome {und c} oligomers act as competitive inhibitors of native {und c}, however, crosslinking of cytochrome {und c} to {und c}-depleted mitoplasts or purified oxidase (with dimethyl suberimidate or hetrobifunctional crosslinking reagents) results in a catalytically inactive complex.

  15. Questions about elastic waves

    CERN Document Server

    Engelbrecht, Jüri


    This book addresses the modelling of mechanical waves by asking the right questions about them and trying to find suitable answers. The questions follow the analytical sequence from elementary understandings to complicated cases, following a step-by-step path towards increased knowledge. The focus is on waves in elastic solids, although some examples also concern non-conservative cases for the sake of completeness. Special attention is paid to the understanding of the influence of microstructure, nonlinearity and internal variables in continua. With the help of many mathematical models for describing waves, physical phenomena concerning wave dispersion, nonlinear effects, emergence of solitary waves, scales and hierarchies of waves as well as the governing physical parameters are analysed. Also, the energy balance in waves and non-conservative models with energy influx are discussed. Finally, all answers are interwoven into the canvas of complexity.

  16. Regenerative Endodontics: Burning Questions. (United States)

    Smith, Anthony J; Cooper, Paul R


    Pulp regeneration and its clinical translation into regenerative endodontic procedures are receiving increasing research attention, leading to significant growth of the published scientific and clinical literature within these areas. Development of research strategies, which consider patient-, clinician-, and scientist-based outcomes, will allow greater focus on key research questions driving more rapid clinical translation. Three key areas of focus for these research questions should include cells, signaling, and infection/inflammation. A translational pathway is envisaged in which clinical approaches are increasingly refined to provide regenerative endodontic protocols that are based on a robust understanding of the physiological processes and events responsible for the normal secretion, structure, and biological behavior of pulpal tissue. Copyright © 2017 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  17. Quantum theory from questions

    CERN Document Server

    Hoehn, Philipp A


    We reconstruct the explicit formalism of qubit quantum theory from elementary rules on an observer's information acquisition. Our approach is purely operational: we consider an observer O interrogating a system S with binary questions and define S's state as O's `catalogue of knowledge' about S; no ontic assumptions are necessary. From the rules we derive the state spaces for N qubits and show that (a) they coincide with the set of density matrices over N qubit Hilbert spaces; (b) states evolve unitarily under the group $\\rm{PSU}(2^N)$ according to the von Neumann evolution equation; and (c) the binary questions by means of which O interrogates the systems corresponds to projective measurements on Pauli operators with outcome probabilities given by the Born rule. Besides offering a novel conceptual perspective on qubit quantum theory, the reconstruction also unravels new structural insights. Namely, we show that, in a quadratic information measure, (d) qubits satisfy informational complementarity inequalities...

  18. A role for NADPH oxidase in antigen presentation

    Directory of Open Access Journals (Sweden)

    Gail J Gardiner


    Full Text Available The nicotinamide adenine dinucleotide phosphate (NADPH oxidase expressed in phagocytes is a multi-subunit enzyme complex that generates superoxide (O2.-. This radical is an important precursor of hydrogen peroxide (H2O2 and other reactive oxygen species (ROS needed for microbicidal activity during innate immune responses. Inherited defects in NADPH oxidase give rise to chronic granulomatous disease (CGD, a primary immunodeficiency characterized by recurrent infections and granulomatous inflammation. Interestingly, CGD, CGD carrier status, and oxidase gene polymorphisms have all been associated with autoinflammatory and autoimmune disorders, suggesting a potential role for NADPH oxidase in regulating adaptive immune responses. Here, NADPH oxidase function in antigen processing and presentation is reviewed. NADPH oxidase influences dendritic cell (DC crosspresentation by major histocompatibility complex class I molecules (MHC-I through regulation of the phagosomal microenvironment, while in B lymphocytes, NADPH oxidase alters epitope selection by major histocompatibility complex class II molecules (MHC-II.

  19. Questions to Luce Irigaray


    Ince, Kate


    This article traces the "dialogue" between the work of the philosophers Luce Irigaray and Emmanuel Levinas. It attempts to construct a more nuanced discussion than has been given to date of Irigaray's critique of Levinas, particularly as formulated in 'Questions to Emmanuel Levinas' (Irigaray 1991). It suggests that the concepts of the feminine and of voluptuosity articulated by Levinas have more to contribute to Irigaray's project of an ethics of sexual difference than she herself sometimes ...

  20. 501 reading comprehension questions

    CERN Document Server


    This updated edition offers the most extensive and varied practice for all types of questions students might face on standardized and in-class tests. With this guide, students will learn to develop expert reading strategies, understand how to read faster and with greater comprehension, overcome reading anxiety, and increase appreciation of reading for pleasure. This book's step-by-step approach provides graduated coverage that moves from the basics to more advanced reading.

  1. Eight Questions about Corruption


    Jakob Svensson


    This paper will discuss eight frequently asked questions about public corruption: (1) What is corruption? (2) Which countries are the most corrupt? (3) What are the common characteristics of countries with high corruption? (4) What is the magnitude of corruption? (5) Do higher wages for bureaucrats reduce corruption? (6) Can competition reduce corruption? (7) Why have there been so few (recent) successful attempts to fight corruption? (8) Does corruption adversely affect growth?

  2. Questioning Many Mysteries (United States)

    Martin, Sara F.


    The first section of this memoir queries my formative years. Indirectly I address the question, did my childhood and early years make a difference in my choice of career? Why and how did I begin my journey to becoming a scientist? Did I choose the field of solar astronomy or did circumstances dictate it for me? In the second section, I travel through my work environments and experiences, talking about interactions and aspects of being a scientist that do not appear in our research papers. What parts of my research were happenstances and what parts did I plan? What does it feel like to be on scientific quests? Using examples in my journey, I also turn to questions that have intrigued me throughout my sojourn as a solar astronomer. How do scientific discoveries come about? What factors lead to little discoveries? And what factors lead to major exciting discoveries? Are there timely questions we do not think to ask? How can small, seemingly scattered pieces of knowledge suddenly coalesce into a deeper understanding - what is called the "Aha!" experience - the times when our mental light switches on, and with child-like wonder we behold a "big picture"?

  3. Proton-pumping mechanism of cytochrome c oxidase: a kinetic master-equation approach. (United States)

    Kim, Young C; Hummer, Gerhard


    Cytochrome c oxidase is an efficient energy transducer that reduces oxygen to water and converts the released chemical energy into an electrochemical membrane potential. As a true proton pump, cytochrome c oxidase translocates protons across the membrane against this potential. Based on a wealth of experiments and calculations, an increasingly detailed picture of the reaction intermediates in the redox cycle has emerged. However, the fundamental mechanism of proton pumping coupled to redox chemistry remains largely unresolved. Here we examine and extend a kinetic master-equation approach to gain insight into redox-coupled proton pumping in cytochrome c oxidase. Basic principles of the cytochrome c oxidase proton pump emerge from an analysis of the simplest kinetic models that retain essential elements of the experimentally determined structure, energetics, and kinetics, and that satisfy fundamental physical principles. The master-equation models allow us to address the question of how pumping can be achieved in a system in which all reaction steps are reversible. Whereas proton pumping does not require the direct modulation of microscopic reaction barriers, such kinetic gating greatly increases the pumping efficiency. Further efficiency gains can be achieved by partially decoupling the proton uptake pathway from the active-site region. Such a mechanism is consistent with the proposed Glu valve, in which the side chain of a key glutamic acid shuttles between the D channel and the active-site region. We also show that the models predict only small proton leaks even in the absence of turnover. The design principles identified here for cytochrome c oxidase provide a blueprint for novel biology-inspired fuel cells, and the master-equation formulation should prove useful also for other molecular machines. .

  4. Purine-induced expression of urate oxidase and enzyme activity in Atlantic salmon (Salmo salar). Cloning of urate oxidase liver cDNA from three teleost species and the African lungfish Protopterus annectens. (United States)

    Andersen, Øivind; Aas, Turid S; Skugor, Stanko; Takle, Harald; van Nes, Solveig; Grisdale-Helland, Barbara; Helland, Ståle J; Terjesen, Bendik F


    The peroxisomal enzyme urate oxidase plays a pivotal role in the degradation of purines in both prokaryotes and eukaryotes. However, knowledge about the purine-induced expression of the encoding gene is lacking in vertebrates. These are the first published sequences of fish urate oxidase, which were predicted from PCR amplified liver cDNAs of Atlantic salmon (Salmo salar), Atlantic cod (Gadus morhua), Atlantic halibut (Hippoglossus hippoglossus) and African lungfish (Protopterus annectens). Sequence alignment of different vertebrate urate oxidases revealed amino acid substitutions of putative functional importance in the enzyme of chicken and lungfish. In the adult salmon, expression of urate oxidase mRNA predominated in liver, but was also identified in several nonhepatic organs including brain, but not in skeletal muscle and kidney. Juvenile salmon fed diets containing bacterial protein meal (BPM) rich in nucleic acids showed a significant increase in liver urate oxidase enzyme activity, and urea concentrations in plasma, muscle and liver were elevated. Whereas salmon fed the 18% BPM diet showed a nonsignificant increase in liver mRNA levels of urate oxidase compared with the 0% BPM-fed fish, no further increase in mRNA levels was found in fish receiving 36% BPM. The discrepancy between urate oxidase mRNA and enzyme activity was explained by rapid mRNA degradation or alternatively, post-translational control of the activity. Although variable plasma and liver levels of urate were detected, the substrate increased only slightly in 36% BPM-fed fish, indicating that the uricolytic pathway of Atlantic salmon is intimately regulated to handle high dietary purine levels.

  5. The Art of Asking Questions. (United States)

    Sykes, Rosetta A.


    A rationale is given for the use of questioning techniques and strategies in classroom instruction. B. Bloom's Taxonomy of Educational Objectives is presented as one framework for questions. Five pitfalls, including avoiding vague questions and personal pronouns, are discussed. (CL)

  6. Xanthine oxidase biosensor for monitoring meat spoilage (United States)

    Vanegas, D. C.; Gomes, C.; McLamore, E. S.


    In this study, we have designed an electrochemical biosensor for real-time detection of specific biomarkers of bacterial metabolism related to meat spoilage (hypoxanthine and xanthine). The selective biosensor was developed by assembling a `sandwich' of nanomaterials and enzymes on a platinum-iridium electrode (1.6 mm tip diameter). The materials deposited on the sensor tip include amorphous platinum nanoclusters (i.e. Pt black), reduced graphene oxide, nanoceria, and xanthine oxidase. Xanthine oxidase was encapsulated in laponite hydrogel and used for the biorecognition of hypoxanthine and xanthine (two molecules involved in the rotting of meat by spoilage microorganisms). The developed biosensor demonstrated good electrochemical performance toward xanthine with sensitivity of 2.14 +/- 1.48 μA/mM, response time of 5.2 +/- 1.5 sec, lower detection limit of 150 +/- 39 nM, and retained at least 88% of its activity after 7 days of continuous use.

  7. Plasma diamine oxidase activity in asthmatic children

    Directory of Open Access Journals (Sweden)

    Kyoichiro Toyoshima


    Full Text Available Histamine plays an important role in the development of asthmatic symptoms. Diamine oxidase (DAO histaminase, which inactivates histamine, is located in the intestine and kidney and is released into plasma. Plasma DAO activity in asthmatic children was measured by a recently developed high performance liquid chromatographic method using histamine as the DAO substrate. Diamine oxidase activity was higher in severely asthmatic children than in those with mild asthma. A time course study during the acute exacerbation phase revealed that DAO activity rose during acute asthmatic attacks and then decreased gradually over several days. Although the mechanisms of plasma DAO activity increase during acute asthmatic attacks could not be explained, data showed that plasma DAO activity is an important index of histamine metabolism in asthmatics and may relate to some mechanisms of acute exacerbation of airway inflammation. Consequently, fluctuations in plasma DAO can be used as one of various indices of instability in management of asthma.

  8. Characterization of polyphenol oxidase from plants

    Institute of Scientific and Technical Information of China (English)

    LEI Dongfeng; FENG Yi; JIANG Dazong


    Polyphenol oxidase (PPO) which can mediate browning reaction is a bifunctional copper-containing enzyme encoded by plant nucleolus gene. It usually leads to excessive browning reaction which reduces the coercial profits of fruits and vegetables. In this paper, PPO genes and enzymes in plants are characterized systematically, and the latest progress is reviewed. Some clonings of PPOs genes are reported; the specific temporal and spatial expression pattern of PPOs genes is described; the model of the structure of the precursor form of catechol oxidase is introduced; the possible functions of PPOs in defending against pathogen, wounding, surrounding stress and other inducing factors are demonstrated; the induction and activation of latent PPOs in some plants is elucidated; the scheme of browning inhibition by L-cysteine is clarified; the mechanism of suicide inhibition of latent PPO and kinetic synergism are established. Furthermore, the area for future study is also discussed.

  9. Process characterization of a monoamine oxidase

    DEFF Research Database (Denmark)

    Ramesh, Hemalata; Woodley, John


    .e, on biocatalyst development (e.g. improvement of expression levels), process development (e.g. improved oxygen supply, product removal strategies) or biocatalyst stabilization (e.g. through immobilization or directed evolution). This paper presents a systematic method to identify the bottleneck of a potential...... biocatalytic process using a monoamine oxidase to synthesise an intermediate in the manufacture of a drug for treating Hepatitis C (Telaprevir)....

  10. Imaging Monoamine Oxidase in the Human Brain

    Energy Technology Data Exchange (ETDEWEB)

    Fowler, J. S.; Volkow, N. D.; Wang, G-J.; Logan, Jean


    Positron emission tomography (PET) studies mapping monoamine oxidase in the human brain have been used to measure the turnover rate for MAO B; to determine the minimum effective dose of a new MAO inhibitor drug lazabemide and to document MAO inhibition by cigarette smoke. These studies illustrate the power of PET and radiotracer chemistry to measure normal biochemical processes and to provide information on the effect of drug exposure on specific molecular targets.

  11. The inhibition of monoamine oxidase by esomeprazole



    Virtual screening of a library of drugs has suggested that esomeprazole, the S-enantiomer of omeprazole, may possess binding affinities for the active sites of the monoamine oxidase (MAO) A and B enzymes. Based on this finding, the current study examines the MAO inhibitory properties of esomeprazole. Using recombinant human MAO-A and MAO-B, IC50 values for the inhibition of these enzymes by esomeprazole were experimentally determined. To examine the reversibility of MAO inhibition by esomepra...

  12. Social Epistemology: 5 Questions

    DEFF Research Database (Denmark)

    Social Epistemology: 5 Questions is a collection of interviews with some of the world’s most influential scholars working on social epistemology from a range of disciplinary perspectives. We hear their views on social epistemology; its aim, scope, use, broader intellectual environment, future...... direction, and how the work of the interviewees fits in these respects. Interviews with David Bloor, Cristina Bicchieri, Richard Bradley, Lorraine Code, Hans van Ditmarsch, Miranda Fricker, Steve Fuller, Sanford Goldberg, Alvin Goldman, Philip Kitcher, Martin Kusch, Jennifer Lackey, Helen E. Longino, Philip...

  13. Alternative Medicine (United States)

    ... Involved News About Us Donate In This Section Alternative Medicine en Español email Send this article to a ... Dr. Yvonne Ou on Lifestyle Modifications and Glaucoma Alternative medicine may be defined as non-standard, unconventional treatments ...

  14. Trick questions: cosmopolitan hospitality

    Directory of Open Access Journals (Sweden)

    Eleanor Byrne


    Full Text Available Byrne’s paper consists of two parallel texts. The first explores the limits of cosmopolitanism in practice, taking as its subject the Life in the UK Citizenship Test, inaugurated under the Labour Government in 2005. It argues that the test exemplifies the predicament of all attempts at cosmopolitan hospitality as unconditional welcoming, through a discussion of the relation between questioning and welcoming the stranger. Establishing the relationship between cosmopolitanism and hospitality as envisaged in Derrida’s reading of Kant it asks what kind of cosmopolitan hospitality is either possible or desirable by exploring what Derrida calls the ‘perversions’ inherent in the structures of hospitality. It focuses on the concept of the ‘trick questions’ that the state asks the foreigner observed by Derrida in his reading of The Apology of Socrates; questions that seem to invite answers but foreclose the possibilities of a free response. The second text asks how this logic that Derrida identifies can be pushed or coaxed into new ways of addressing the perceived threats of ‘unconditional’ hospitality through a reading of ‘unconditional hospitality’ as queer in the work of Tove Jansson.

  15. The Gentle Art of Questioning: Writing Great Clicker Questions (United States)

    Chasteen, Stephanie


    How does a teacher use questioning effectively? This workshop will focus on writing those questions that engage students, spark their curiosity, help recap material, give you insight into their thinking, or help them learn critical ideas in physics. We will focus on ``peer instruction'' -- a research-tested method of requiring students to discuss challenging questions with one another. We will investigate the surprising power of multiple-choice questions to achieve critical thinking skills. Finally, we will look at writing questions that align with our goals for students, discuss the elements of effective questions, and practice writing questions and work on improving them.

  16. Questions and Questioning Techniques: A View of Indonesian Students’ Preferences

    Directory of Open Access Journals (Sweden)

    Debora Tri Ragawanti


    Full Text Available This study investigated students’ preference on teacher’s questions and questionings techniques and more importantly on how they could facilitate or impede their learning. The results on teacher’s questioning techniques showed that random nomination was more preferred than pre-arranged format nomination. In addition, techniques of nominating volunteering students and of giving wait-time were disliked by most student-respondents. As for types of question, the yes/no question was favored by most of the respondents. Different from the yes/no question, the number of respondents leaning forward to the analysis question, questions about fact of life, and questions to state opinion did not show a significant difference from the number of those leaning against the same questions.

  17. Adipogenesis-related increase of semicarbazide-sensitive amine oxidase and monoamine oxidase in human adipocytes. (United States)

    Bour, Sandy; Daviaud, Danièle; Gres, Sandra; Lefort, Corinne; Prévot, Danielle; Zorzano, Antonio; Wabitsch, Martin; Saulnier-Blache, Jean-Sébastien; Valet, Philippe; Carpéné, Christian


    A strong induction of semicarbazide-sensitive amine oxidase (SSAO) has previously been reported during murine preadipocyte lineage differentiation but it remains unknown whether this emergence also occurs during adipogenesis in man. Our aim was to compare SSAO and monoamine oxidase (MAO) expression during in vitro differentiation of human preadipocytes and in adipose and stroma-vascular fractions of human fat depots. A human preadipocyte cell strain from a patient with Simpson-Golabi-Behmel syndrome was first used to follow amine oxidase expression during in vitro differentiation. Then, human preadipocytes isolated from subcutaneous adipose tissues were cultured under conditions promoting ex vivo adipose differentiation and tested for MAO and SSAO expression. Lastly, human adipose tissue was separated into mature adipocyte and stroma-vascular fractions for analyses of MAO and SSAO at mRNA, protein and activity levels. Both SSAO and MAO were increased from undifferentiated preadipocytes to lipid-laden cells in all the models: 3T3-F442A and 3T3-L1 murine lineages, human SGBS cell strain or human preadipocytes in primary culture. In human subcutaneous adipose tissue, the adipocyte-enriched fraction exhibited seven-fold higher amine oxidase activity and contained three- to seven-fold higher levels of mRNAs encoded by MAO-A, MAO-B, AOC3 and AOC2 genes than the stroma-vascular fraction. MAO-A and AOC3 genes accounted for the majority of their respective MAO and SSAO activities in human adipose tissue. Most of the SSAO and MAO found in adipose tissue originated from mature adipocytes. Although the mechanism and role of adipogenesis-related increase in amine oxidase expression remain to be established, the resulting elevated levels of amine oxidase activities found in human adipocytes may be of potential interest for therapeutic intervention in obesity.

  18. From Questions to Queries

    Directory of Open Access Journals (Sweden)

    M. Drlík


    Full Text Available The extension of (Internet databases forceseveryone to become more familiar with techniques of datastorage and retrieval because users’ success often dependson their ability to pose right questions and to be able tointerpret their answers. University programs pay moreattention to developing database programming skills than todata exploitation skills. To educate our students to become“database users”, the authors intensively exploit supportivetools simplifying the production of database elements astables, queries, forms, reports, web pages, and macros.Videosequences demonstrating “standard operations” forcompleting them have been prepared to enhance out-ofclassroomlearning. The use of SQL and other professionaltools is reduced to the cases when the wizards are unable togenerate the intended construct.

  19. The social question revisited

    DEFF Research Database (Denmark)

    Kenneth, Petersson; Olsson, Ulf; Krejsler, John B.


    of social and political unrest, poverty and lack of morality. In the name of European Union social integration is thus organized differently as compared to former times. There are, nevertheless similarities. In both cases educational systems become key arenas for integrating social groups......The focus of this chapter is the re-installation of the social question as a historical practice. The purpose is to investigate how historic figures return and are applied in contemporary political discourses, more precisely in the context of education, education policy and teacher education...... and raising the level of knowledge and competences in society. Higher education, in this case teacher education, is supposed to develop potentials of individuals and maximize “their contribution to a sustainable and democratic knowledge-based society.” Consequently; student cohorts should reflect...

  20. The questions for Machiavelli

    Directory of Open Access Journals (Sweden)

    Lakićević Dragan D.


    Full Text Available The main intention of the work that deals with the Nicolo Machiavelli thought is to point out the obvious paradox between the high political goal and the legitimating of all possible means for its realization. Are evil deeds inevitable in the sphere of politics and under what circumstances the immorality contained in political acts could be transformed into common good? The text asks additional questions such as about the accomplishments of ambitious political projects, the relationship among the ideologist and the representative of political power, the transformations of the means into the ends, the use of violence and indoctrination in political acts, revolutionary and evolutionary political methods, etc. The author claims that political technologies recommended by Machiavelli basically haven't diminished, but have taken on more modern and more adequate forms.

  1. The Deflection Question (United States)

    Greenberg, A. H.; Nesvold, E.; van Heerden, E.; Erasmus, N.; Marchis, F.


    On 15 February, 2013, a 15 m diameter asteroid entered the Earth's atmosphere over Russia. The resulting shockwave injured nearly 1500 people, and incurred 33 million (USD) in infrastructure damages. The Chelyabinsk meteor served as a forceful demonstration of the threat posed to Earth by the hundreds of potentially hazardous objects (PHOs) that pass near the Earth every year. Although no objects have yet been discovered on an impact course for Earth, an impact is virtually statistically guaranteed at some point in the future. While many impactor deflection technologies have been proposed, humanity has yet to demonstrate the ability to divert an impactor when one is found. Developing and testing any single proposed technology will require significant research time and funding. This leaves open an obvious question - towards which technologies should funding and research be directed, in order to maximize our preparedness for when an impactor is eventually found? To help answer this question, we have created a detailed framework for analyzing various deflection technologies and their effectiveness. Using an n-body integrator (REBOUND), we have simulated the attempted deflections of a population of Earth-impacting objects with a variety of velocity perturbations (∂Vs), and measured the effects that these perturbations had on impact probability. We then mapped the ∂Vs applied in the orbital simulations to the technologies capable of achieving those perturbations, and analyzed which set of technologies would be most effective at preventing a PHO from impacting the earth. As a final step, we used the results of these simulations to train a machine learning algorithm. This algorithm, combined with a simulated PHO population, can predict which technologies are most likely to be needed. The algorithm can also reveal which impactor observables (mass, spin, orbit, etc.) have the greatest effect on the choice of deflection technology. These results can be used as a tool to

  2. Some Criticisms against Exam Questions: To What Extent Can Mythic, Romantic and Philosophical Questions Be Used? (United States)

    Seker, Hasan


    In the present study, some of the pre-service teachers' criticisms against their exams were investigated. Moreover, as an alternative, to what extent philosophical, romantic and mythic questions could be used was also looked at. The study group consists of 117 pre-service teachers from the classroom teacher education. In the study, it was…

  3. Automatic Chinese Factual Question Generation (United States)

    Liu, Ming; Rus, Vasile; Liu, Li


    Question generation is an emerging research area of artificial intelligence in education. Question authoring tools are important in educational technologies, e.g., intelligent tutoring systems, as well as in dialogue systems. Approaches to generate factual questions, i.e., questions that have concrete answers, mainly make use of the syntactical…

  4. Pathological changes in platelet histamine oxidases in atopic eczema

    Directory of Open Access Journals (Sweden)

    Reinhold Kiehl


    Full Text Available Increased plasma histamine levels were associated with significantly lowered diamine and type B monoamine oxidase activities in platelet-rich plasma of atopic eczema (AE patients. The diamine oxidase has almost normal cofactor levels (pyridoxal phosphate and Cu2+ but the cofactor levels for type B monoamine oxidase (flavin adenine dinucleotide and Fe2+ are lowered. The biogenic amines putrescine, cadaverine, spermidine, spermine, tyramine and serotonin in the sera, as well as dopamine and epinephrine in EDTA-plasma were found to be normal. It is unlikely, therefore, that these amines are responsible for the decreased activities of monoamine and diamine oxidase in these patients. The most likely causative factors for the inhibition of the diamine oxidase are nicotine, alcohol, food additives and other environmental chemicals, or perhaps a genetic defect of the diamine oxidase.

  5. The nature of CuA in cytochrome c oxidase


    Stevens, Tom H.; Martin, Craig T.; Wang, Hsin; Brudvig, Gary W.; Scholes, Charles P.; Chan, Sunney I.


    The isolation and purification of yeast cytochrome c oxidase is described. Characterization of the purified protein indicates that it is spectroscopically identical with cytochrome c oxidase isolated from beef heart. Preparations of isotopically substituted yeast cytochrome c oxidase are obtained incorporating [1,3-15N2]histidine or [beta,beta- 2H2]cysteine. Electron paramagnetic resonance and electron nuclear double resonance spectra of the isotopically substituted proteins identify unambigu...

  6. [The sanitary question in the modernity-postmodernity debate]. (United States)

    Iriart, C; Spinelli, H


    This work analyzes the sanitary question in the modernity-postmodernity debate. Such analyses are performed form a philosophical position that states the crisis of Modernity and questions the ideological twist that to itself propitiates postmodernity, shutting out questioning views or visions. It propitiates an alternative view of politics, thinking of it from the potency plane and giving a role to the subject in the decision of producing transformations.

  7. Nanodesign: some basic questions

    CERN Document Server

    Schommers, Wolfram


    There is no doubt that nanoscience will be the dominant direction for technology in this century, and that this science will influence our lives to a large extent as well as open completely new perspectives on all scientific and technological disciplines. To be able to produce optimal nanosystems with tailor-made properties, it is necessary to analyze and construct such systems in advance by adequate theoretical and computational methods. Since we work in nanoscience and nanotechnology at the ultimate level, we have to apply the basic laws of physics. What methods and tools are relevant here? The book gives an answer to this question. The background of the theoretical methods and tools is critically discussed, and also the world view on which these physical laws are based. Such a debate is not only of academic interest but is of highly general concern, and this is because we constantly move in nanoscience and nanotechnology between two extreme poles, between infinite life and total destruction . On the one ...

  8. NADPH oxidase: an enzyme for multicellularity? (United States)

    Lalucque, Hervé; Silar, Philippe


    Multicellularity has evolved several times during the evolution of eukaryotes. One evolutionary pressure that permits multicellularity relates to the division of work, where one group of cells functions as nutrient providers and the other in specialized roles such as defence or reproduction. This requires signalling systems to ensure harmonious development of multicellular structures. Here, we show that NADPH oxidases are specifically present in organisms that differentiate multicellular structures during their life cycle and are absent from unicellular life forms. The biochemical properties of these enzymes make them ideal candidates for a role in intercellular signalling.

  9. NADH/NADPH Oxidase and Vascular Function. (United States)

    Griendling, K K; Ushio-Fukai, M


    The vascular NADH/NADPH oxidase has been shown to be the major source of superoxide in the vessel wall. Recent work has provided insight into its structure and activity in vascular cells. This enzyme is involved in both vascular smooth muscle hypertrophy and in some forms of impaired endothelium-dependent relaxation. Because oxidative stress in general participates in the pathogenesis of hypertension and atherosclerosis, the enzymes that produce reactive oxygen species may be important determinants of the course of vascular disease. (Trends Cardiovasc Med 1997;7:301-307). © 1997, Elsevier Science Inc.


    Felton, L D


    1. A simple technique is described for studying the oxidase action of bacteria by means of the oxidation of p-aminoleucomalachite green. 2. It is shown that pneumococci under aerobic conditions produced an oxidase when grown on suitable medium. The sera of any of seven different animal species constitute such a medium, the degree of oxidation by the pneumococcus depending upon the animal from which the serum was taken-rat, guinea pig, rabbit, horse, man, cat, and chicken in order of diminishing suitability. 3. Conditions favoring the oxidation of p-aminoleucomalachite green by a single strain of pneumococci are: the presence of a slight amount of hemoglobin, dextrose, H ion concentration on the add side, and heating of fresh serum for 30 minutes at 56 degrees C. Conditions preventing the oxidation are: sterilized meat infusion, 1 per cent peptone, plain broth, a high concentration of hemoglobin, and absence of oxygen. In a quantitative fashion, meat infusion, 1 per cent peptone, and plain broth interfere with the suitability of serum as a substratum of oxidase production by the pneumococcus. 4. Twenty-three microbic species were studied with reference to oxidative power. They were grown upon 10 per cent horse serum, with and without dextrose, upon 10 per cent guinea pig serum, and upon plain broth. Only three of the twenty-three gave evidence of oxidative power as tested by p-aminoleucomalachite green; namely, the pneumococcus, Streptococcus viridans, and Streptococcus haemolyticus. Among the strains, of these three pneumococci gave the most intense reaction, after which Streptococcus viridans and Streptococcus haemolyticus follow in the order named, but with a noticeable variation among the different strains of Streptococcus haemolyticus. 5. Hemolytic streptococci of human and bovine origin were studied. The only variation in the type of reaction was manifested by the streptococci of milk and cheese origin. Strains from these sources showed definitely the least

  11. Questioning Strategies of English Teachers

    Institute of Scientific and Technical Information of China (English)



    AMY.B.M.TSUI thinks that most of the interactive learning starts with the teachers ’questioning. Richard and Lock-art (1994) think that the proper questioning can help the students to acquire the second language. Jin Chuanbao ( 1997) even thinks that the questioning process of the teachers should become the core of the class. Though almost all of the teachers are ques-tioning, they know little about the questioning strategies. In this case, it is urgent to study this subject. The present study reveals some problems of four teachers’questioning strategies in Junior Middle School. I hope some helpful ideas can be found in the thesis.

  12. A question of authority

    Energy Technology Data Exchange (ETDEWEB)

    Morgan, Earl W.(BATTELLE (PACIFIC NW LAB))


    A Question of Authority. This article deals with a certain scenario and several reviewers are to give their opinion. This one is in regards to - Suspending an IACUC approved animal use activity is about the last thing a research institution wants to do. Consider the predicament that the Great Eastern University IACUC faced when Dr. Janet Jenkins, the Attending Veterinarian, suspended all animal use activity on an approved protocol of Dr. Roy Maslo. Jenkins had the IACUCs authority to temporarily suspend a protocol, subject to review by a quorum of the full committee. She alleged that Maslo used mice from his breeding colony, not purchased rats, to begin a new study. Jenkins saw Maslos technicians bringing mouse cages to a procedure room and setting up for a minor survival surgery. She asked them to wait until she clarified things as she felt confident that the protocol called for rats. She called Maslo and asked him if the study had been approved for mice, to which he responded affirmatively. Still not feeling quite assured, she went to her office, reviewed the protocol, and found only rat studies described. She also called the IACUC office to see if there were any approved amendments which she may not have received, and was told that there were none. By the time she returned, one procedure was completed. Understandably upset, she informed the technicians and Maslo that any further activity on the protocol was suspended until the issue was resolved. Jenkins informed the IACUC chairman who in turned called an emergency meeting of the committee.

  13. Multicopper oxidase-1 orthologs from diverse insect species have ascorbate oxidase activity. (United States)

    Peng, Zeyu; Dittmer, Neal T; Lang, Minglin; Brummett, Lisa M; Braun, Caroline L; Davis, Lawrence C; Kanost, Michael R; Gorman, Maureen J


    Members of the multicopper oxidase (MCO) family of enzymes can be classified by their substrate specificity; for example, ferroxidases oxidize ferrous iron, ascorbate oxidases oxidize ascorbate, and laccases oxidize aromatic substrates such as diphenols. Our previous work on an insect multicopper oxidase, MCO1, suggested that it may function as a ferroxidase. This hypothesis was based on three lines of evidence: RNAi-mediated knock down of Drosophila melanogaster MCO1 (DmMCO1) affects iron homeostasis, DmMCO1 has ferroxidase activity, and DmMCO1 has predicted iron binding residues. In our current study, we expanded our focus to include MCO1 from Anopheles gambiae, Tribolium castaneum, and Manduca sexta. We verified that MCO1 orthologs have similar expression profiles, and that the MCO1 protein is located on the basal surface of cells where it is positioned to oxidize substrates in the hemolymph. In addition, we determined that RNAi-mediated knock down of MCO1 in A. gambiae affects iron homeostasis. To further characterize the enzymatic activity of MCO1 orthologs, we purified recombinant MCO1 from all four insect species and performed kinetic analyses using ferrous iron, ascorbate and two diphenols as substrates. We found that all of the MCO1 orthologs are much better at oxidizing ascorbate than they are at oxidizing ferrous iron or diphenols. This result is surprising because ascorbate oxidases are thought to be specific to plants and fungi. An analysis of three predicted iron binding residues in DmMCO1 revealed that they are not required for ferroxidase or laccase activity, but two of the residues (His374 and Asp380) influence oxidation of ascorbate. These two residues are conserved in MCO1 orthologs from insects and crustaceans; therefore, they are likely to be important for MCO1 function. The results of this study suggest that MCO1 orthologs function as ascorbate oxidases and influence iron homeostasis through an unknown mechanism.

  14. Alternative wastewatersystems

    DEFF Research Database (Denmark)

    Dyck-Madsen, Søren; Hoffmann, Birgitte; Gabriel, Søren


    The report:-  Communicates experiences from Swedish buildings from the establishment and running of alternative wastewater systems. Communicates pictures of alternative buildings and wastewater systems in Sweden. Gives a short evaluation of the performance and the sustainability of the systems....

  15. Antibiotic Resistance Questions and Answers (United States)

    ... on the Farm Get Smart About Antibiotics Week Antibiotic Resistance Questions and Answers Language: English (US) Español ( ... Many ear infections Top of Page Questions about Antibiotic Resistance Examples of How Antibiotic Resistance Spreads Click for ...

  16. Enzymatic Characterization and In Vivo Function of Five Terminal Oxidases in Pseudomonas aeruginosa


    Arai, Hiroyuki; Kawakami, Takuro; Osamura, Tatsuya; Hirai, Takehiro; Sakai, Yoshiaki; Ishii, Masaharu


    The ubiquitous opportunistic pathogen Pseudomonas aeruginosa has five aerobic terminal oxidases: bo3-type quinol oxidase (Cyo), cyanide-insensitive oxidase (CIO), aa3-type cytochrome c oxidase (aa3), and two cbb3-type cytochrome c oxidases (cbb3-1 and cbb3-2). These terminal oxidases are differentially regulated under various growth conditions and are thought to contribute to the survival of this microorganism in a wide variety of environmental niches. Here, we constructed multiple mutant str...

  17. The Questions of Liberal Education (United States)

    Arcilla, Rene V.


    There is a certain kind of liberal educator who bases his or her practice on a particular attitude toward the "Big Questions." The questions of fundamental literacy in K-12 education, or of expertise in vocational and professional education, may be just as important, but they are seen as quite different in kind. Indeed, the questions of liberal…

  18. The Hermeneutics of Educational Questioning (United States)

    Bingham, Charles


    This article looks at the practice of educational questioning using the philosophical hermeneutics of Hans-Georg Gadamer. It first looks at questions and statements from a hermeneutic perspective, demonstrating some of the differences and similarities between the two. It then details Gadamer's notion of the "true question", asking whether it is…

  19. Disjunctive questions, intonation, and highlighting

    NARCIS (Netherlands)

    F. Roelofsen; S. van Gool


    This paper examines how intonation affects the interpretation of disjunctive questions. The semantic effect of a question is taken to be three-fold. First, it raises an issue. In the tradition of inquisitive semantics, we model this by assuming that a question proposes several possible updates of th

  20. Does Anyone Have Any Questions? (United States)

    Finkelstein, Judith M.; Ritter, Virginia F.

    The purpose of this study was to determine if answering a child's question with a question produces further analytical questioning by the child. A sample of 80 children in nursery-kindergarten, first, second and third grades (ages ranging from 4-9 years) were divided into two groups. An abstract painting by Kandinsky was shown individually to each…

  1. The Hermeneutics of Educational Questioning (United States)

    Bingham, Charles


    This article looks at the practice of educational questioning using the philosophical hermeneutics of Hans-Georg Gadamer. It first looks at questions and statements from a hermeneutic perspective, demonstrating some of the differences and similarities between the two. It then details Gadamer's notion of the "true question", asking whether it is…

  2. Children Who Question Their Heterosexuality (United States)

    Carver, Priscilla R.; Egan, Susan K.; Perry, David G.


    Many gay, lesbian, and bisexual adults report a period of childhood sexual questioning--an uneasy questioning of their heterosexuality brought on by same-sex attractions and motivating same-sex sexual exploration. This article evaluates hypotheses about the correlates, causes, and consequences of childhood sexual questioning. Participants were 182…

  3. Laboratory-evolved vanillyl-alcohol oxidase produces natural vanillin

    NARCIS (Netherlands)

    Heuvel, van den R.H.H.; Berg, van den W.A.M.; Rovida, S.; Berkel, van W.J.H.


    The flavoenzyme vanillyl-alcohol oxidase was subjected to random mutagenesis to generate mutants with enhanced reactivity to creosol (2-methoxy-4-methylphenol). The vanillyl-alcohol oxidase-mediated conversion of creosol proceeds via a two-step process in which the initially formed vanillyl alcohol

  4. Endothelins and NADPH oxidases in the cardiovascular system. (United States)

    Dammanahalli, Karigowda J; Sun, Zhongjie


    1. The endothelin (ET) system and NADPH oxidase play important roles in the regulation of cardiovascular function, as well as in the pathogenesis of hypertension and other cardiovascular diseases. 2. Endothelins activate NADPH oxidases and thereby increase superoxide production, resulting in oxidative stress and cardiovascular dysfunction. Thus, NADPH oxidases may mediate the role of endothelins in some cardiovascular diseases. However, the role of reactive oxygen species (ROS) in mediating ET-induced vasoconstriction and cardiovascular disease remains under debate, as evidenced by conflicting reports from different research teams. Conversely, activation of NADPH oxidase can stimulate ET secretion via ROS generation, which further enhances the cardiovascular effects of NADPH oxidase. However, little is known about how ROS activate the endothelin system. It seems that the relationship between ET-1 and ROS may vary with cardiovascular disorders. 3. Endothelins activate NADPH oxidase via the ET receptor-proline-rich tyrosine kinase-2 (Pyk2)-Rac1 pathway. Rac1 is an important regulator of NADPH oxidase. There is ample evidence supporting direct stimulation by Rac1 of NADPH oxidase activity. In addition, Rac1-induced cardiomyocyte hypertrophy is mediated by the generation of ROS.

  5. Priming and activation of NADPH oxidases in plants and animals. (United States)

    Canton, Johnathan; Grinstein, Sergio


    In mammals, engagement of Toll-like receptors by microbe-associated molecular patterns enhances the responsiveness of NADPH oxidases. Two recent papers report a similar 'priming' mechanism for the plant oxidase RbohD. Despite lacking structural homology, the functional parallels between plants and animals reveal that a common regulatory logic arose by convergent evolution. Copyright © 2014 Elsevier Ltd. All rights reserved.

  6. Xanthine Oxidase Inhibitory Activity of a Plectranthus saccatus aqueous extract

    Directory of Open Access Journals (Sweden)

    Caldeira F


    Full Text Available Gout is a disease with high prevalence in developed countries, resulting from the deposition of uric acid crystals in various locations, particularly at the joints. The pharmacotherapeutic approach to chronic gout essentially consists of administration of uric acid-lowering agents. The main mechanism of action of these agents is the inhibition of xanthine oxidase (XO, the enzyme responsible for the formation of uric acid. The therapeutic alternatives available for this purpose are limited, thus justifying the interest of the discovery of potential new uric acidlowering drugs. In this regard, an aqueous extract of the plant Plectranthus saccatus has been studied for its ability to inhibit XO. The composition of the extract was determined by HPLC and rosmarinic acid was identified as the major constituent. Both the extract and rosmarinic acid have demonstrated the ability to inhibit the production of uric acid by interfering with XO activity. The results obtained herein support the continuation of the study of their uric acid-lowering properties in cell-based and in vivo models to further explore their potential in gout therapy.

  7. Questions That This Monograph Raises

    Directory of Open Access Journals (Sweden)



    Full Text Available What is the ideal combination of increased investment in biomedical research by sponsors with least conflict of interest in researchers?What should interested parties do to refurbish pharma's image?Is a voluntary moratorium on pharma spending practically possible?How to resolve the basic schism between profit driven industry and welfare driven profession of medicine?How can CPGs be salvaged from ulterior influence?Can guidelines on guidelines help?Is cost-effectiveness of CPGs a practical solution?Is it possible to grade therapies as Most, Moderate and Least Cost Effective?How can Disease specific Foundations be salvaged from sponsor's influence?How can Journals ensure scientific integrity of conflicted authors and nullify untoward influence of sponsors?Are reports like the Task Force on individual and institutional conflicts of interest of the AAMC followed more in their breach?Are the revised ICMJE guidelines adequate to stub conflicted research?Editorials make pious announcements. Do they really influence what researchers do?How do we protect the interests of human research subjects?Do Best Practice Guidelines and Good Publication Practices really help?Marketability is the name of the game, not usefulness. How do we ensure the latter, even as the former is forwarded?How can we ensure effective traditional therapies remain in use even as new ones are forwarded?How do we ensure non-pharmacological therapies are also forwarded?How can the legitimate thrust of biological psychiatry be encouraged while also forwarding non-pharmacological approaches in psychiatry?What can be done so pharma just cannot consider questionable means as an attractive alternative?How do we expedite scientific self-correction while causing least harm to patients and research subjects?

  8. Modulation of NADPH oxidase activity by known uraemic retention solutes

    DEFF Research Database (Denmark)

    Schulz, Anna Marta; Terne, Cindy; Jankowski, Vera


    as the strongest inhibitor of NADPH oxidase (90% of DPI inhibition). Surprisingly, none of the uraemic retention solutes we investigated was found to increase NADPH oxidase activity. Furthermore, plasma from patients with CKD-5D before dialysis caused significantly higher inhibitory effect on NADPH oxidase...... inhibitory effect on NADPH oxidase activity in the presence of plasma from patients with CKD-5D after dialysis compared with before dialysis, we investigated the effect of 48 known and commercially available uraemic retention solutes on the enzymatic activity of NADPH oxidase. METHODS: Mononuclear leucocytes...... isolated from buffy coats of healthy volunteers were isolated, lysed and incubated with NADH in the presence of plasma from healthy controls and patients with CKD-5D. Furthermore, the leucocytes were lysed and incubated in the presence of uraemic retention solute of interest and diphenyleneiodonium...

  9. The NADH oxidase-Prx system in Amphibacillus xylanus. (United States)

    Niimura, Youichi


    Amphibacillus NADH oxidase belongs to a growing new family of peroxiredoxin-linked oxidoreductases including alkyl hydroperoxide reductase F (AhpF). Like AhpF it displays extremely high hydroperoxide reductase activity in the presence of a Prx, thus making up the NADH oxidase-Prx system. The NADH oxidase primarily catalyzes the reduction of oxygen by NADH to form H2O2, while the Prx immediately reduces H2O2 (or ROOH) to water (or ROH). Consequently, the NADH oxidase-Prx system catalyzes the reduction of both oxygen and hydrogen peroxide to water with NADH as the preferred electron donor. The NADH oxidase-Prx system is widely distributed in aerobically growing bacteria lacking a respiratory chain and catalase, and plays an important role not only in scavenging hydroperoxides but also in regenerating NAD in these bacteria.

  10. Arts and Techniques of Questioning

    Institute of Scientific and Technical Information of China (English)



      Most learners learn English from teachers in classrooms, therefore, classroom instruction is very important. However, teachers’questioning plays a very significant part in classroom teaching. It is not only an important part of classroom interaction but also an effective way of learning second language as wel. On the one hand, it is through question and answer exchange that teachers interactwithstudents.Onthe other hand, it isalsothroughinteractionwiththeirteachersand peers that ESLlearners learn the target language. Teachers’ aims of questioning, question types, teachers’modification of questions, the wait-time, the feedback and the assessment providedby teacherswil affectclassroom interaction.

  11. Can multiple-choice questions simulate free-response questions?

    CERN Document Server

    Lin, Shih-Yin


    We discuss a study to evaluate the extent to which free-response questions could be approximated by multiple-choice equivalents. Two carefully designed research-based multiple-choice questions were transformed into a free-response format and administered on the final exam in a calculus-based introductory physics course. The original multiple-choice questions were administered in another similar introductory physics course on final exam. Findings suggest that carefully designed multiple-choice questions can reflect the relative performance of the free-response questions while maintaining the benefits of ease of grading and quantitative analysis, especially if the different choices in the multiple-choice questions are weighted to reflect the different levels of understanding that students display.

  12. Increased xanthine oxidase-related ROS production and TRPV1 synthesis preceding DOMS post-eccentric exercise in rats. (United States)

    Retamoso, Leandro T; Silveira, Mauro E P; Lima, Frederico D; Busanello, Guilherme L; Bresciani, Guilherme; Ribeiro, Leandro R; Chagas, Pietro M; Nogueira, Cristina W; Braga, Ana Claudia M; Furian, Ana Flávia; Oliveira, Mauro S; Fighera, Michele R; Royes, Luiz Fernando F


    It is well-known that unaccustomed exercise, especially eccentric exercise, is associated to delayed onset muscle soreness (DOMS). Whether DOMS is associated with reactive oxygen species (ROS) and the transient receptor potential vanilloid 1 (TRPV1) is still an open question. Thus, the aim of this study was to investigate the association between TRPV1 and xanthine oxidase-related ROS production in muscle and DOMS after a bout of eccentric exercise. Male Wistar rats performed a downhill running exercise on a treadmill at a -16° tilt and a constant speed for 90min (5min/bout separated by 2min of rest). Mechanical allodynia and grip force tests were performed before and 1, 3, 6, 9, 12, 24, 48 and 72h after the downhill running. Biochemical assays probing oxidative stress, purine degradation, xanthine oxidase activity, Ca(2+) ATPase activity and TRPV1 protein content were performed in gastrocnemius muscle at 12, 24, and 48h after the downhill running. Our statistical analysis showed an increase in mechanical allodynia and a loss of strength after the downhill running. Similarly, an increase in carbonyl, xanthine oxidase activity, uric acid levels and TRPV1 immunoreactivity were found 12h post-exercise. On the other hand, Ca(2+) ATPase activity decreased in all analyzed times. Our results suggest that a possible relationship between xanthine oxidase-related ROS and TRPV1 may exist during the events preceding eccentric exercise-related DOMS. Copyright © 2016 Elsevier Inc. All rights reserved.

  13. Do clinicians use more question marks? (United States)

    Otte, Willem M; van’t Klooster, Maryse A; van Diessen, Eric; Leijten, Frans SS; Sander, Josemir W


    Objective To quantify the use of question marks in titles of published studies. Design and setting Literature review. Participants All Pubmed publications between 1 January 2013 and 31 December 2013 with an available abstract. Papers were classified as being clinical when the search terms clin*, med* or patient* were found anywhere in the paper’s title, abstract or the journal’s name. Other papers were considered controls. As a verification, clinical journals were compared to non-clinical journals in two different approaches. Also, 50 highest impact journals were explored for publisher group dependent differences. Main outcome measure Total number of question marks in titles. Results A total of 368,362 papers were classified as clinical and 596,889 as controls. Clinical papers had question marks in 3.9% (95% confidence interval 3.8–4.0%) of titles and other papers in 2.3% (confidence interval 2.3–2.3%; p < 0.001). These findings could be verified for clinical journals compared to non-clinical journals. Different percentages between four publisher groups were found (p < 0.01). Conclusion We found more question marks in titles of clinical papers than in other papers. This could suggest that clinicians often have a question-driven approach to research and scientists in more fundamental research a hypothesis-driven approach. An alternative explanation is that clinicians like catchy titles. Publishing groups might have pro- and anti-question mark policies. PMID:26085937

  14. NADPH Oxidases in Chronic Liver Diseases

    Directory of Open Access Journals (Sweden)

    Joy X. Jiang


    Full Text Available Oxidative stress is a common feature observed in a wide spectrum of chronic liver diseases including viral hepatitis, alcoholic, and nonalcoholic steatohepatitis. The nicotinamide adenine dinucleotide phosphate (NADPH oxidases (NOXs are emerging as major sources of reactive oxygen species (ROS. Several major isoforms are expressed in the liver, including NOX1, NOX2, and NOX4. While the phagocytic NOX2 has been known to play an important role in Kupffer cell and neutrophil phagocytic activity and inflammation, the nonphagocytic NOX homologues are increasingly recognized as key enzymes in oxidative injury and wound healing. In this review, we will summarize the current advances in knowledge on the regulatory pathways of NOX activation, their cellular distribution, and their role in the modulation of redox signaling in liver diseases.

  15. Glucose oxidase immobilization onto carbon nanotube networking

    CERN Document Server

    Karachevtsev, V A; Zarudnev, E S; Karachevtsev, M V; Leontiev, V S; Linnik, A S; Lytvyn, O S; Plokhotnichenko, A M; Stepanian, S G


    When elaborating the biosensor based on single-walled carbon nanotubes (SWNTs), it is necessary to solve such an important problem as the immobilization of a target biomolecule on the nanotube surface. In this work, the enzyme (glucose oxidase (GOX)) was immobilized on the surface of a nanotube network, which was created by the deposition of nanotubes from their solution in 1,2-dichlorobenzene by the spray method. 1-Pyrenebutanoic acid succinimide ester (PSE) was used to form the molecular interface, the bifunctional molecule of which provides the covalent binding with the enzyme shell, and its other part (pyrene) is adsorbed onto the nanotube surface. First, the usage of such a molecular interface leaves out the direct adsorption of the enzyme (in this case, its activity decreases) onto the nanotube surface, and, second, it ensures the enzyme localization near the nanotube. The comparison of the resonance Raman (RR) spectrum of pristine nanotubes with their spectrum in the PSE environment evidences the creat...

  16. Characterization of Glucose Oxidase from Penicillium notatum

    Directory of Open Access Journals (Sweden)

    Nabeela Saleem


    Full Text Available In the present study glucose oxidase (GOD has been isolated from a culture filtrate of Penicillium notatum. The enzyme was purified by ammonium sulphate precipitation, diethylaminoethyl (DEAE cellulose ion-exchange chromatography and Sephadex gel filtration. This protocol gave 16.47-fold purification and 25 % recovery of the enzyme. The optimum pH and temperature for the activity were 5.4 and 45 °C, respectively. The Km and vmax values for the enzyme were 10.5 mM and 456 U/mg, respectively. A detailed kinetic study of thermal inactivation was carried out. Both enthalpy of activation (ΔH* and entropy of activation (ΔS* decreased at higher temperatures. Moreover, free energy of denaturation (ΔG* increased at higher temperature, making the enzyme thermally stable. A possible explanation for the thermal inactivation of GOD at higher temperatures is also discussed.

  17. Steady-state oxidation of cholesterol catalyzed by cholesterol oxidase in lipid bilayer membranes on platinum electrodes

    Energy Technology Data Exchange (ETDEWEB)

    Bokoch, Michael P.; Devadoss, Anando; Palencsar, Mariela S.; Burgess, James D


    Cholesterol oxidase is immobilized in electrode-supported lipid bilayer membranes. Platinum electrodes are initially modified with a self-assembled monolayer of thiolipid. A vesicle fusion method is used to deposit an outer leaflet of phospholipids onto the thiolipid monolayer forming a thiolipid/lipid bilayer membrane on the electrode surface. Cholesterol oxidase spontaneously inserts into the electrode-supported lipid bilayer membrane from solution and is consequently immobilized to the electrode surface. Cholesterol partitions into the membrane from buffer solutions containing cyclodextrin. Cholesterol oxidase catalyzes the oxidation of cholesterol by molecular oxygen, forming hydrogen peroxide as a product. Amperometric detection of hydrogen peroxide for continuous solution flow experiments are presented, where flow was alternated between cholesterol solution and buffer containing no cholesterol. Steady-state anodic currents were observed during exposures of cholesterol solutions ranging in concentration from 10 to 1000 {mu}M. These data are consistent with the Michaelis-Menten kinetic model for oxidation of cholesterol as catalyzed by cholesterol oxidase immobilized in the lipid bilayer membrane. The cholesterol detection limit is below 1 {mu}M for cholesterol solution prepared in buffered cyclodextrin. The response of the electrodes to low density lipoprotein solutions is increased upon addition of cyclodextrin. Evidence for adsorption of low density lipoprotein to the electrode surface is presented.

  18. Extra virgin olive oil rich in polyphenols modulates VEGF-induced angiogenic responses by preventing NADPH oxidase activity and expression. (United States)

    Calabriso, Nadia; Massaro, Marika; Scoditti, Egeria; D'Amore, Simona; Gnoni, Antonio; Pellegrino, Mariangela; Storelli, Carlo; De Caterina, Raffaele; Palasciano, Giuseppe; Carluccio, Maria Annunziata


    Previous studies have shown the antiinflammatory, antioxidant and antiangiogenic properties by pure olive oil polyphenols; however, the effects of olive oil phenolic fraction on the inflammatory angiogenesis are unknown. In this study, we investigated the effects of the phenolic fraction (olive oil polyphenolic extract, OOPE) from extra virgin olive oil and related circulating metabolites on the VEGF-induced angiogenic responses and NADPH oxidase activity and expression in human cultured endothelial cells. We found that OOPE (1-10 μg/ml), at concentrations achievable nutritionally, significantly reduced, in a concentration-dependent manner, the VEGF-induced cell migration, invasiveness and tube-like structure formation through the inhibition of MMP-2 and MMP-9. OOPE significantly (Polive oil, with high polyphenol content, decreased VEGF-induced NADPH oxidase activity and Nox4 expression, as well as, MMP-9 expression, as compared with fasting control serum. Overall, native polyphenols and serum metabolites of extra virgin olive oil rich in polyphenols are able to lower the VEGF-induced angiogenic responses by preventing endothelial NADPH oxidase activity and decreasing the expression of selective NADPH oxidase subunits. Our results provide an alternative mechanism by which the consumption of olive oil rich in polyphenols may account for a reduction of oxidative stress inflammatory-related sequelae associated with chronic degenerative diseases.

  19. Portability of oxidase domains in nonribosomal peptide synthetase modules. (United States)

    Schneider, Tanya L; Walsh, Christopher T


    Oxazole and thiazole rings are present in numerous nonribosomal peptide natural products. Oxidase domains are responsible for catalyzing the oxidation of thiazolines and oxazolines to yield fully aromatic heterocycles. Unlike most domains, the placement of oxidase domains within assembly line modules varies. Noting this tolerance, we investigated the portability of an oxidase domain to a heterologous assembly line. The epimerase domain of PchE, involved in pyochelin biosynthesis, was replaced with the oxidase domain from MtaD, involved in myxothiazol biosynthesis. The chimeric module was expressed in soluble form as a flavin mononucleotide-containing flavoprotein. The functionality of the inserted oxidase domain was assayed within PchE and in transfer of the growing siderophore acyl chain from PchE to the next downstream module. While pyochelin-like product release was not observed downstream, the robust activity of the transplanted oxidase domain and the ability of the chimeric module to produce an advanced intermediate bound to the synthetase underscore the possibility of future engineering within nonribosomal peptide synthetase pathways using oxidase domains.

  20. Instance-Based Question Answering (United States)


    included factoid questions grouped around a set of target entities. For example, for the target entity “ Franz Kafka ”, associated questions included: “Where...from Franz Schubert. A year later he did on Dec 5th.”). Depending on the task, an answer extractor may identify very small, factoid candidates which...IWP): Paraphrase Ac- quisition and Applications, 2003. [55] A. Ittycheriah, M. Franz , and S. Roukos. Ibm’s statistical question answering system - trec

  1. And the next question is powerful questions for sticky moments

    CERN Document Server

    Alexander, Rachel


    A very practical and easy to use book of 3,000+ powerful questions, forming part of every coach''s / manager''s toolkit; it enables you to easily find key questions in some of the most distinctive areas of coaching, such as confidence, communications & leadership.

  2. Commonly Asked Questions in Thermodynamics

    CERN Document Server

    Assael, Marc J


    Have you ever had a question that keeps persisting and for which you cannot find a clear answer? Is the question seemingly so "simple" that the problem is glossed over in most resources, or skipped entirely? CRC Press/Taylor and Francis is pleased to introduce Commonly Asked Questions in Thermodynamics, the first in a new series of books that address the questions that frequently arise in today's major scientific and technical disciplines. Designed for a wide audience, from students and researchers to practicing professionals in related areas, the books are organized in a user friend

  3. Direct regulation of cytochrome c oxidase by calcium ions.

    Directory of Open Access Journals (Sweden)

    Tatiana Vygodina

    Full Text Available Cytochrome c oxidase from bovine heart binds Ca(2+ reversibly at a specific Cation Binding Site located near the outer face of the mitochondrial membrane. Ca(2+ shifts the absorption spectrum of heme a, which allowed previously to determine the kinetics and equilibrium characteristics of the binding. However, no effect of Ca(2+ on the functional characteristics of cytochrome oxidase was revealed earlier. Here we report that Ca(2+ inhibits cytochrome oxidase activity of isolated bovine heart enzyme by 50-60% with Ki of ∼1 µM, close to Kd of calcium binding with the oxidase determined spectrophotometrically. The inhibition is observed only at low, but physiologically relevant, turnover rates of the enzyme (∼10 s(-1 or less. No inhibitory effect of Ca(2+ is observed under conventional conditions of cytochrome c oxidase activity assays (turnover number >100 s(-1 at pH 8, which may explain why the effect was not noticed earlier. The inhibition is specific for Ca(2+ and is reversed by EGTA. Na(+ ions that compete with Ca(2+ for binding with the Cation Binding Site, do not affect significantly activity of the enzyme but counteract the inhibitory effect of Ca(2+. The Ca(2+-induced inhibition of cytochrome c oxidase is observed also with the uncoupled mitochondria from several rat tissues. At the same time, calcium ions do not inhibit activity of the homologous bacterial cytochrome oxidases. Possible mechanisms of the inhibition are discussed as well as potential physiological role of Ca(2+ binding with cytochrome oxidase. Ca(2+- binding at the Cation Binding Site is proposed to inhibit proton-transfer through the exit part of the proton conducting pathway H in the mammalian oxidases.

  4. CotA, a multicopper oxidase from Bacillus pumilus WH4, exhibits manganese-oxidase activity.

    Directory of Open Access Journals (Sweden)

    Jianmei Su

    Full Text Available Multicopper oxidases (MCOs are a family of enzymes that use copper ions as cofactors to oxidize various substrates. Previous research has demonstrated that several MCOs such as MnxG, MofA and MoxA can act as putative Mn(II oxidases. Meanwhile, the endospore coat protein CotA from Bacillus species has been confirmed as a typical MCO. To study the relationship between CotA and the Mn(II oxidation, the cotA gene from a highly active Mn(II-oxidizing strain Bacillus pumilus WH4 was cloned and overexpressed in Escherichia coli strain M15. The purified CotA contained approximately four copper atoms per molecule and showed spectroscopic properties typical of blue copper oxidases. Importantly, apart from the laccase activities, the CotA also displayed substantial Mn(II-oxidase activities both in liquid culture system and native polyacrylamide gel electrophoresis. The optimum Mn(II oxidase activity was obtained at 53°C in HEPES buffer (pH 8.0 supplemented with 0.8 mM CuCl2. Besides, the addition of o-phenanthroline and EDTA both led to a complete suppression of Mn(II-oxidizing activity. The specific activity of purified CotA towards Mn(II was 0.27 U/mg. The Km, Vmax and kcat values towards Mn(II were 14.85±1.17 mM, 3.01×10(-6±0.21 M·min(-1 and 0.32±0.02 s(-1, respectively. Moreover, the Mn(II-oxidizing activity of the recombinant E. coli strain M15-pQE-cotA was significantly increased when cultured both in Mn-containing K liquid medium and on agar plates. After 7-day liquid cultivation, M15-pQE-cotA resulted in 18.2% removal of Mn(II from the medium. Furthermore, the biogenic Mn oxides were clearly observed on the cell surfaces of M15-pQE-cotA by scanning electron microscopy. To our knowledge, this is the first report that provides the direct observation of Mn(II oxidation with the heterologously expressed protein CotA, Therefore, this novel finding not only establishes the foundation for in-depth study of Mn(II oxidation mechanisms, but also offers

  5. Questioning Questions: Elementary Teachers' Adaptations of Investigation Questions Across the Inquiry Continuum (United States)

    Biggers, Mandy


    Questioning is a central practice in science classrooms. However, not every question translates into a "good" science investigation. Questions that drive science investigations can be provided by many sources including the teacher, the curriculum, or the student. The variations in the source of investigation questions were explored in this study. A dataset of 120 elementary science classroom videos and associated lesson plans from 40 elementary teachers (K-5) across 21 elementary school campuses were scored on an instrument measuring the amount of teacher-direction or student-direction of the lessons' investigation questions. Results indicated that the investigation questions were overwhelmingly teacher directed in nature, with no opportunities for students to develop their own questions for investigation. This study has implications for researchers and practitioners alike, calling attention to the teacher-directed nature of investigation questions in existing science curriculum materials, and the need for teacher training in instructional strategies to adapt their existing curriculum materials across the continuum of teacher-directed and student-directed investigation questions. Teachers need strategies for adapting the teacher-directed questions provided in their existing curriculum materials in order to allow students the opportunity to engage in this essential scientific practice.

  6. Multilayered polyelectrolyte microcapsules: interaction with the enzyme cytochrome C oxidase. (United States)

    Pastorino, Laura; Dellacasa, Elena; Noor, Mohamed R; Soulimane, Tewfik; Bianchini, Paolo; D'Autilia, Francesca; Antipov, Alexei; Diaspro, Alberto; Tofail, Syed A M; Ruggiero, Carmelina


    Cell-sized polyelectrolyte capsules functionalized with a redox-driven proton pump protein were assembled for the first time. The interaction of polyelectrolyte microcapsules, fabricated by electrostatic layer-by-layer assembly, with cytochrome c oxidase molecules was investigated. We found that the cytochrome c oxidase retained its functionality, that the functionalized microcapsules interacting with cytochrome c oxidase were permeable and that the permeability characteristics of the microcapsule shell depend on the shell components. This work provides a significant input towards the fabrication of an integrated device made of biological components and based on specific biomolecular functions and properties.

  7. Multilayered polyelectrolyte microcapsules: interaction with the enzyme cytochrome C oxidase.

    Directory of Open Access Journals (Sweden)

    Laura Pastorino

    Full Text Available Cell-sized polyelectrolyte capsules functionalized with a redox-driven proton pump protein were assembled for the first time. The interaction of polyelectrolyte microcapsules, fabricated by electrostatic layer-by-layer assembly, with cytochrome c oxidase molecules was investigated. We found that the cytochrome c oxidase retained its functionality, that the functionalized microcapsules interacting with cytochrome c oxidase were permeable and that the permeability characteristics of the microcapsule shell depend on the shell components. This work provides a significant input towards the fabrication of an integrated device made of biological components and based on specific biomolecular functions and properties.

  8. Secretory production of an FAD cofactor-containing cytosolic enzyme (sorbitol-xylitol oxidase from Streptomyces coelicolor) using the twin-arginine translocation (Tat) pathway of Corynebacterium glutamicum. (United States)

    Scheele, Sandra; Oertel, Dan; Bongaerts, Johannes; Evers, Stefan; Hellmuth, Hendrik; Maurer, Karl-Heinz; Bott, Michael; Freudl, Roland


    Carbohydrate oxidases are biotechnologically interesting enzymes that require a tightly or covalently bound cofactor for activity. Using the industrial workhorse Corynebacterium glutamicum as the expression host, successful secretion of a normally cytosolic FAD cofactor-containing sorbitol-xylitol oxidase from Streptomyces coelicolor was achieved by using the twin-arginine translocation (Tat) protein export machinery for protein translocation across the cytoplasmic membrane. Our results demonstrate for the first time that, also for cofactor-containing proteins, a secretory production strategy is a feasible and promising alternative to conventional intracellular expression strategies.

  9. Probability and Statistics: 5 Questions

    DEFF Research Database (Denmark)

    Probability and Statistics: 5 Questions is a collection of short interviews based on 5 questions presented to some of the most influential and prominent scholars in probability and statistics. We hear their views on the fields, aims, scopes, the future direction of research and how their work fits...

  10. Alternative Veier

    DEFF Research Database (Denmark)

    Kruse, Tove Elisabeth; Salamonsen, Anita

    reflektioner omkring patienters brug af og erfaringer med alternativ behandling. Patientorganisationer, organisatoner for alternative behandlere og organisationer for læger og medicinstuderende har læst bogens patienthistorier og deres perspektiver lægges frem. Til slut i bogen diskuteres betydningen af de...

  11. Growing Alternatives

    DEFF Research Database (Denmark)

    Bagger-Petersen, Mai Corlin


    From 2014, Anhui Province will pilot a reform of the residential land market in China, thus integrating rural Anhui in the national housing market. In contrast, artist and activist Ou Ning has proposed the Bishan time money currency, intending to establish an alternative economic circuit in Bishan...

  12. Magnetostrictive Alternator (United States)

    Dyson, Rodger; Bruder, Geoffrey


    This innovation replaces the linear alternator presently used in Stirling engines with a continuous-gradient, impedance-matched, oscillating magnetostrictive transducer that eliminates all moving parts via compression, maintains high efficiency, costs less to manufacture, reduces mass, and eliminates the need for a bearing system. The key components of this new technology are the use of stacked magnetostrictive materials, such as Terfenol-D, under a biased magnetic and stress-induced compression, continuous-gradient impedance-matching material, coils, force-focusing metallic structure, and supports. The acoustic energy from the engine travels through an impedancematching layer that is physically connected to the magnetostrictive mass. Compression bolts keep the structure under compressive strain, allowing for the micron-scale compression of the magnetostrictive material and eliminating the need for bearings. The relatively large millimeter displacement of the pressure side of the impedance-matching material is reduced to micron motion, and undergoes stress amplification at the magnetostrictive interface. The alternating compression and expansion of the magnetostrictive material creates an alternating magnetic field that then induces an electric current in a coil that is wound around the stack. This produces electrical power from the acoustic pressure wave and, if the resonant frequency is tuned to match the engine, can replace the linear alternator that is commonly used.

  13. Alternative Treatments (United States)

    ... triglyceride (fat) produced by processing coconut oil or palm kernel oil. The body breaks down caprylic acid into substances called “ketone bodies.” The theory behind Axona is that the ketone bodies derived from caprylic acid may provide an alternative energy source for brain cells that have lost ...

  14. The Value Question in Metaphysics. (United States)

    Kahane, Guy


    Much seems to be at stake in metaphysical questions about, for example, God, free will or morality. One thing that could be at stake is the value of the universe we inhabit-how good or bad it is. We can think of competing philosophical positions as describing possibilities, ways the world might turn out to be, and to which value can be assigned. When, for example, people hope that God exists, or fear that we do not possess free will, they express attitudes towards these possibilities, attitudes that presuppose answers to questions about their comparative value. My aim in this paper is to distinguish these evaluative questions from related questions with which they can be confused, to identify structural constraints on their proper pursuit, and to address objections to their very coherence. Answers to such evaluative questions offer one measure of the importance of philosophical disputes.

  15. Questions

    DEFF Research Database (Denmark)

    Larsen, Steen Nepper


    Bidrag til festskrift til Jesper Hoffmeyer i anledning af hans 70 års dag i Don Favineau, Paul Cobley & Kalevi Kull (eds.): "A More Developed Sign. Interpreting the Work of Jesper Hoffmeyer". Antologien udg. som særnummer af Tartu Semiotics Library Nr. 10 og mit bidrag forefindes på p. 217-220....

  16. Xanthine oxidase in human skeletal muscle following eccentric exercise

    DEFF Research Database (Denmark)

    Hellsten, Ylva; Frandsen, Ulrik; Orthenblad, N.


    1. The present study tested the hypothesis that the level of xanthine oxidase is elevated in injured human skeletal muscle in association with inflammatory events. Seven male subjects performed five bouts of strenuous one-legged eccentric exercise. Muscle biopsies from both the exercised and the ......1. The present study tested the hypothesis that the level of xanthine oxidase is elevated in injured human skeletal muscle in association with inflammatory events. Seven male subjects performed five bouts of strenuous one-legged eccentric exercise. Muscle biopsies from both the exercised...... the increase in xanthine oxidase in the muscle there were no detectable changes in the levels of muscle malondialdehyde or in plasma antioxidant capacity up to 4 days post-exercise. 5. It is concluded that eccentric exercise leads to an increased level of xanthine oxidase in human muscle and that the increase...

  17. Aldehyde-induced xanthine oxidase activity in raw milk. (United States)

    Steffensen, Charlotte L; Andersen, Henrik J; Nielsen, Jacob H


    In the present study, the aldehyde-induced pro-oxidative activity of xanthine oxidase was followed in an accelerated raw milk system using spin-trap electron spin resonance (ESR) spectroscopy. The aldehydes acetaldehyde, propanal, hexanal, trans-2-hexenal, trans-2-heptenal, trans-2-nonenal, and 3-methyl-2-butenal were all found to initiate radical reactions when added to milk. Formation of superoxide through aldehyde-induced xanthine oxidase activity is suggested as the initial reaction, as all tested aldehydes were shown to trigger superoxide formation in an ultrahigh temperature (UHT) milk model system with added xanthine oxidase. It was found that addition of aldehydes to milk initially increased the ascorbyl radical concentration with a subsequent decay due to ascorbate depletion, which renders the formation of superoxide in milk with added aldehyde. The present study shows for the first time potential acceleration of oxidative events in milk through aldehyde-induced xanthine oxidase activity.

  18. Activation of the neutrophil NADPH oxidase by Aspergillus fumigatus. (United States)

    Boyle, Keith B; Stephens, Len R; Hawkins, Phillip T


    Upon infection of the respiratory system with the fungus Aspergillus fumigatus various leukoctytes, in particular neutrophils, are recruited to the lung to mount an immune response. Neutrophils respond by both phagocytosing conidia and mediating extracellular killing of germinated, invasive hyphae. Of paramount importance to an appropriate immune response is the neutrophil NADPH oxidase enzyme, which mediates the production of various reactive oxygen species (ROS). This is evidenced by the acute sensitivity of both oxidase-deficient humans and mice to invasive aspergillosis. Herein we briefly review the mechanisms and functions of oxidase activation and discuss our recent work identifying at least some of the important players in hyphal-induced oxidase activation and neutrophil function. Among these we define the phosphoinositide 3-kinase enzyme and the regulatory protein Vav to be of critical importance and allude to a kinase-independent role for Syk.

  19. Phrasal Paraphrase Based Question Reformulation for Archived Question Retrieval. (United States)

    Zhang, Yu; Zhang, Wei-Nan; Lu, Ke; Ji, Rongrong; Wang, Fanglin; Liu, Ting


    Lexical gap in cQA search, resulted by the variability of languages, has been recognized as an important and widespread phenomenon. To address the problem, this paper presents a question reformulation scheme to enhance the question retrieval model by fully exploring the intelligence of paraphrase in phrase-level. It compensates for the existing paraphrasing research in a suitable granularity, which either falls into fine-grained lexical-level or coarse-grained sentence-level. Given a question in natural language, our scheme first detects the involved key-phrases by jointly integrating the corpus-dependent knowledge and question-aware cues. Next, it automatically extracts the paraphrases for each identified key-phrase utilizing multiple online translation engines, and then selects the most relevant reformulations from a large group of question rewrites, which is formed by full permutation and combination of the generated paraphrases. Extensive evaluations on a real world data set demonstrate that our model is able to characterize the complex questions and achieves promising performance as compared to the state-of-the-art methods.

  20. Phrasal Paraphrase Based Question Reformulation for Archived Question Retrieval.

    Directory of Open Access Journals (Sweden)

    Yu Zhang

    Full Text Available Lexical gap in cQA search, resulted by the variability of languages, has been recognized as an important and widespread phenomenon. To address the problem, this paper presents a question reformulation scheme to enhance the question retrieval model by fully exploring the intelligence of paraphrase in phrase-level. It compensates for the existing paraphrasing research in a suitable granularity, which either falls into fine-grained lexical-level or coarse-grained sentence-level. Given a question in natural language, our scheme first detects the involved key-phrases by jointly integrating the corpus-dependent knowledge and question-aware cues. Next, it automatically extracts the paraphrases for each identified key-phrase utilizing multiple online translation engines, and then selects the most relevant reformulations from a large group of question rewrites, which is formed by full permutation and combination of the generated paraphrases. Extensive evaluations on a real world data set demonstrate that our model is able to characterize the complex questions and achieves promising performance as compared to the state-of-the-art methods.

  1. Hypothetical constructs, hypothetical questions, and the expert witness. (United States)

    Brodsky, Stanley L; Titcomb, Caroline; Sams, David M; Dickson, Kara; Benda, Yves


    Professor John Henry Wigmore in 1940 described the hypothetical question as an intolerable obstruction of truth. Since that time, the nature and application of the hypothetical question in the courtroom, as well as responses to this line of questioning during expert testimony, have been sources of controversy. Governed by legal philosophical foundations, the hypothetical construct addresses what there is, in a general sense, and what can or ought to be. Alexy (2004) has described the nature of legal philosophy as the epistemological question of what we can know. This article begins by examining the philosophical underpinnings, legal parameters, and teaching purposes of posing hypothetical queries. A social-psychological backdrop for the use of hypothetical questions is then discussed followed by a broader discussion of the hypothetical question's role in court procedures. This paper identifies hypothetical questions used in court as devices to elicit information, or as predictions that potentially change underlying factual interpretations of evidence. In particular, on cross examination hypothetical questions seek to make opposing experts assume facts that are incongruent with their conclusions or opinions. Sometimes in these situations, experts are led to re-evaluate opinions based on alternative understandings of events and behaviors. Thus, this paper's final aim is to explore a foundational understanding of hypothetical questions asked of expert witnesses with special reference to mental health issues. Options for responding to hypothetical questions on the stand are considered along the dimensions of assertiveness-passivity, compliance-resistance, and possible redefinitions of the hypothetical issues.

  2. Inventory control: cytochrome c oxidase assembly regulates mitochondrial translation. (United States)

    Mick, David U; Fox, Thomas D; Rehling, Peter


    Mitochondria maintain genome and translation machinery to synthesize a small subset of subunits of the oxidative phosphorylation system. To build up functional enzymes, these organellar gene products must assemble with imported subunits that are encoded in the nucleus. New findings on the early steps of cytochrome c oxidase assembly reveal how the mitochondrial translation of its core component, cytochrome c oxidase subunit 1 (Cox1), is directly coupled to the assembly of this respiratory complex.

  3. Regulation of the NADPH Oxidase RBOHD During Plant Immunity



    Pathogen recognition induces the production of reactive oxygen species (ROS) by NADPH oxidases in both plants and animals. ROS have direct antimicrobial properties, but also serve as signaling molecules to activate further immune outputs. However, ROS production has to be tightly controlled to avoid detrimental effects on host cells, but yet must be produced in the right amount, at the right place and at the right time upon pathogen perception. Plant NADPH oxidases belong to the respiratory b...

  4. Explaining errors in children's questions. (United States)

    Rowland, Caroline F


    The ability to explain the occurrence of errors in children's speech is an essential component of successful theories of language acquisition. The present study tested some generativist and constructivist predictions about error on the questions produced by ten English-learning children between 2 and 5 years of age. The analyses demonstrated that, as predicted by some generativist theories [e.g. Santelmann, L., Berk, S., Austin, J., Somashekar, S. & Lust. B. (2002). Continuity and development in the acquisition of inversion in yes/no questions: dissociating movement and inflection, Journal of Child Language, 29, 813-842], questions with auxiliary DO attracted higher error rates than those with modal auxiliaries. However, in wh-questions, questions with modals and DO attracted equally high error rates, and these findings could not be explained in terms of problems forming questions with why or negated auxiliaries. It was concluded that the data might be better explained in terms of a constructivist account that suggests that entrenched item-based constructions may be protected from error in children's speech, and that errors occur when children resort to other operations to produce questions [e.g. Dabrowska, E. (2000). From formula to schema: the acquisition of English questions. Cognitive Liguistics, 11, 83-102; Rowland, C. F. & Pine, J. M. (2000). Subject-auxiliary inversion errors and wh-question acquisition: What children do know? Journal of Child Language, 27, 157-181; Tomasello, M. (2003). Constructing a language: A usage-based theory of language acquisition. Cambridge, MA: Harvard University Press]. However, further work on constructivist theory development is required to allow researchers to make predictions about the nature of these operations.

  5. Questions About Venus after Venus Express (United States)

    Limaye, Sanjay


    The observations from Venus Express for nearly 13 Venus years or 26 solar days from April 2006 till 27 November 2014. Earlier, Venus has been explored by fly-by spacecraft, orbiters, descending probes, landers and floating balloons. These data have been supplemented by many ground based observations at reflected solar wavelengths, short and long wave infrared, millimeter to radio waves. Venus Express added significantly to the collection that will continue to be examined for understanding the planet's atmosphere and continuing analysis will inform us about new facets of the atmosphere and raise new questions. Inter-comparison of the measurements have been able to provide a general idea of the global atmosphere. However, re-visiting these observations also raises some questions about the atmosphere that have not received much attention lately but deserve to be explored and considered for future measurements. These questions are about the precise atmospheric composition in the deep atmosphere, the atmospheric state in the lower atmosphere, the static stability of the lower atmosphere, the clouds and hazes, the nature of the ultraviolet absorber(s) in the cloud layer, and wind speed and direction near the surface from equator to the pole, interaction between the atmosphere and the solid planet. The answers to these questions are important for a better understanding of Venus, its weather and climate and how the climate has evolved. The questions include: (i) What are the implications of the supercritical state of the two primary constituents of the Venus atmosphere - carbon dioxide and nitrogen in the lower atmosphere? (ii) Is the Venus (lower) atmosphere well mixed? (iii) What determines the observed alternating stable and unstable layers (static stability) in the lower atmosphere? (iv) What causes the contrasts seen in reflected sunlight which are largest at ultraviolet wavelengths and very muted at other visible wavelengths? (v) what causes the morning -afternoon

  6. Traditional Assessment versus Alternative Assessment. (United States)

    Belle, Dana

    The purpose of this study was to determine whether a teacher can use one type of assessment to evaluate students' abilities fairly. The question is whether or not alternative assessment strategies are necessary to meet students' individual needs. The research, conducted with 28 fifth-grade students, compared their traditional and alternative…

  7. Leflunomide, a Reversible Monoamine Oxidase Inhibitor. (United States)

    Petzer, Jacobus P; Petzer, Anél


    A screening study aimed at identifying inhibitors of the enzyme, monoamine oxidase (MAO), among clinically used drugs have indicated that the antirheumatic drug, leflunomide, is an inhibitor of both MAO isoforms. Leflunomide inhibits human MAO-A and MAO-B and exhibits IC50 values of 19.1 μM and 13.7 μM, respectively. The corresponding Ki values are 17.7 μM (MAO-A) and 10.1 μM (MAO-B). Dialyses of mixtures of the MAO enzymes and leflunomide show that inhibition of the MAOs by leflunomide is reversible. The principal metabolite of leflunomide, teriflunomide (A77 1726), in contrast is not an MAO inhibitor. This study concludes that, although leflunomide is only moderately potent as an MAO inhibitor, isoxazole derivatives may represent a general class of MAO inhibitors and this heterocycle may find application in MAO inhibitor design. In this respect, MAO inhibitors are used in the clinic for the treatment of depressive illness and Parkinson's disease, and are under investigation as therapy for certain types of cancer, Alzheimer's disease and age-related impairment of cardiac function.

  8. Origin and evolution of lysyl oxidases. (United States)

    Grau-Bové, Xavier; Ruiz-Trillo, Iñaki; Rodriguez-Pascual, Fernando


    Lysyl oxidases (LOX) are copper-dependent enzymes that oxidize primary amine substrates to reactive aldehydes. The best-studied role of LOX enzymes is the remodeling of the extracellular matrix (ECM) in animals by cross-linking collagens and elastin, although intracellular functions have been reported as well. Five different LOX enzymes have been identified in mammals, LOX and LOX-like (LOXL) 1 to 4, showing a highly conserved catalytic carboxy terminal domain and more divergence in the rest of the sequence. Here we have surveyed a wide selection of genomes in order to infer the evolutionary history of LOX. We identified LOX proteins not only in animals, but also in many other eukaryotes, as well as in bacteria and archaea - which reveals a pre-metazoan origin for this gene family. LOX genes expanded during metazoan evolution resulting in two superfamilies, LOXL2/L3/L4 and LOX/L1/L5. Considering the current knowledge on the function of mammalian LOX isoforms in ECM remodeling, we propose that LOXL2/L3/L4 members might have preferentially been involved in making cross-linked collagen IV-based basement membrane, whereas the diversification of LOX/L1/L5 forms contributed to chordate/vertebrate-specific ECM innovations, such as elastin and fibronectin. Our work provides a novel view on the evolution of this family of enzymes.

  9. Modular assembly of yeast cytochrome oxidase. (United States)

    McStay, Gavin P; Su, Chen Hsien; Tzagoloff, Alexander


    Previous studies of yeast cytochrome oxidase (COX) biogenesis identified Cox1p, one of the three mitochondrially encoded core subunits, in two high-molecular weight complexes combined with regulatory/assembly factors essential for expression of this subunit. In the present study we use pulse-chase labeling experiments in conjunction with isolated mitochondria to identify new Cox1p intermediates and place them in an ordered pathway. Our results indicate that before its assimilation into COX, Cox1p transitions through five intermediates that are differentiated by their compositions of accessory factors and of two of the eight imported subunits. We propose a model of COX biogenesis in which Cox1p and the two other mitochondrial gene products, Cox2p and Cox3p, constitute independent assembly modules, each with its own complement of subunits. Unlike their bacterial counterparts, which are composed only of the individual core subunits, the final sequence in which the mitochondrial modules associate to form the holoenzyme may have been conserved during evolution.

  10. Study of Drug Metabolism by Xanthine Oxidase

    Directory of Open Access Journals (Sweden)

    Lizhou Sun


    Full Text Available In this work, we report the studies of drug metabolism by xanthine oxidase (XOD with electrochemical techniques. Firstly, a pair of stable, well-defined and quasi-reversible oxidation/reduction peaks is obtained with the formal potential at −413.1 mV (vs. SCE after embedding XOD in salmon sperm DNA membrane on the surface of pyrolytic graphite electrode. Then, a new steady peak can be observed at −730 mV (vs. SCE upon the addition of 6-mercaptopurine (6-MP to the electrochemical system, indicating the metabolism of 6-MP by XOD. Furthermore, the chronoamperometric response shows that the current of the catalytic peak located at −730 mV increases with addition of 6-MP in a concentration-dependent manner, and the increase of the chronoamperometric current can be inhibited by an XOD inhibitor, quercetin. Therefore, our results prove that XOD/DNA modified electrode can be efficiently used to study the metabolism of 6-MP, which may provide a convenient approach for in vitro studies on enzyme-catalyzed drug metabolism.

  11. Monoamine oxidase inhibitory activities of heterocyclic chalcones. (United States)

    Minders, Corné; Petzer, Jacobus P; Petzer, Anél; Lourens, Anna C U


    Studies have shown that natural and synthetic chalcones (1,3-diphenyl-2-propen-1-ones) possess monoamine oxidase (MAO) inhibition activities. Of particular importance to the present study is a report that a series of furanochalcones acts as MAO-B selective inhibitors. Since the effect of heterocyclic substitution, other than furan (and more recently thiophene, piperidine and quinoline) on the MAO inhibitory properties of the chalcone scaffold remains unexplored, the aim of this study was to synthesise and evaluate further heterocyclic chalcone analogues as inhibitors of the human MAOs. For this purpose, heterocyclic chalcone analogues that incorporate pyrrole, 5-methylthiophene, 5-chlorothiophene and 6-methoxypyridine substitution were examined. Seven of the nine synthesised compounds exhibited IC50 values chalcones are reversible and competitive MAO inhibitors. 4h, however, may exhibit tight-binding to MAO-B, a property linked to its thiophene moiety. We conclude that high potency chalcones such as 4h represent suitable leads for the development of MAO-B inhibitors for the treatment of Parkinson's disease and possibly other neurodegenerative disorders.

  12. Solar physics: Dynamo theory questioned (United States)

    Charbonneau, Paul


    Observations of X-ray emission -- a diagnostic tool for the mechanisms driving stellar magnetic fields -- from four cool stars call into question accepted models of magnetic-field generation in the Sun and stars. See Letter p.526

  13. LGBT Caregiving: Frequently Asked Questions (United States)

    ... FCA - A A + A You are here Home LGBT Caregiving: Frequently Asked Questions Order this publication Printer- ... service or organization is open to working with LGBT families? Kudos to you for managing to “go ...

  14. Frequent Questions About Universal Waste (United States)

    Frequent questions such as Who is affected by the universal waste regulations? What is “mercury-containing equipment”? How are waste batteries managed under universal waste? How are waste pesticides managed under universal waste?

  15. HPV Vaccine - Questions and Answers (United States)

    ... and Media Resources News Newsletters Events Redirect for HPV Vaccine FAQ Recommend on Facebook Tweet Share Compartir ... to the address below. File Formats Help: How ...

  16. Question Inventory on Tobacco (QIT) (United States)

    U.S. Department of Health & Human Services — 1965, 1966, 1970, 1974-2017. Centers for Disease Control and Prevention (CDC). Office on Smoking and Health (OSH). Tobacco-Related Survey Questions. The QIT is a...

  17. Questions to Ask Your Doctor (United States)

    ... to MBCN Contact Us Questions to ask your doctor Medical appointments can be stressful. To better deal ... for you. If diagnosed by your primary care physician Where do you send your metastatic patients for ...

  18. Questions Students Ask: Beta Decay. (United States)

    Koss, Jordan; Hartt, Kenneth


    Answers a student's question about the emission of a positron from a nucleus. Discusses the problem from the aspects of the uncertainty principle, beta decay, the Fermi Theory, and modern physics. (YP)

  19. Climate Leadership Awards Frequent Questions (United States)

    Provides answers to frequently asked questions regarding the Climate Leadership Awards, sponsored by EPA's Center for Corporate Climate Leadership with co-sponsorship from the Center for Climate and Energy Solutions and The Climate Registry.

  20. Frequently Asked Questions about Pharmacogenomics (United States)

    ... Care Specific Genetic Disorders Frequently Asked Questions About Pharmacogenomics Enlarge What is pharmacogenomics? What might pharmacogenomics mean ... page, you will need Adobe Reader. What is pharmacogenomics? Pharmacogenomics uses information about a person's genetic makeup, ...

  1. Question Inventory on Tobacco (QIT) (United States)

    U.S. Department of Health & Human Services — 1965, 1966, 1970, 1974-2015, 2017. Centers for Disease Control and Prevention (CDC). Office on Smoking and Health (OSH). Survey Questions (Tobacco Use). The QIT is...

  2. Key Questions in Thoracic Surgery


    Subotic, Dragan R.


    This 1000-page textbook encompasses much more than the title suggests. In fact, the title “Key questions in thoracic surgery and pulmonology” would be more fitting. The specific format of the book, with precise questions and evidence-based, but equally clear answers covering all relevant fields of pulmonology and thoracic surgery, makes this 40-chapter book a “must read” not only for residents, but also for senior pulmonologists and thoracic surgeons.

  3. A simple assay for mammalian spermine oxidase: a polyamine catabolic enzyme implicated in drug response and disease. (United States)

    Goodwin, Andrew C; Murray-Stewart, Tracy R; Casero, Robert A


    Spermine oxidase (SMO), the most recently characterized polyamine metabolic enzyme, catalyzes the direct back-conversion of spermine to spermidine in an FAD-dependent reaction that also yields the byproducts hydrogen peroxide (H(2)O(2)) and 3-aminopropanal. These metabolites, particularly H(2)O(2), have been implicated in cytotoxic cellular responses to specific antitumor polyamine analogs, as well as in the inflammation-associated generation of DNA damage. This chapter describes a rapid, sensitive, and inexpensive method for the chemiluminescent measurement of SMO (or alternatively, N (1)-acetyl polyamine oxidase, APAO) enzyme activity in cultured cell lysates, without the need for radioactive reagents or the use of high performance liquid chromatography (HPLC). Specifically, H(2)O(2) production by SMO is coupled to chemiluminescence generated by the horseradish peroxidase-catalyzed oxidation of luminol. Detailed protocols for preparation of reagents, harvesting cell lysates, generation of a standard curve, assaying of samples, and calculation of SMO enzyme activity are presented.

  4. Protein Electrochemistry: Questions and Answers. (United States)

    Fourmond, V; Léger, C

    This chapter presents the fundamentals of electrochemistry in the context of protein electrochemistry. We discuss redox proteins and enzymes that are not photoactive. Of course, the principles described herein also apply to photobioelectrochemistry, as discussed in later chapters of this book. Depending on which experiment is considered, electron transfer between proteins and electrodes can be either direct or mediated, and achieved in a variety of configurations: with the protein and/or the mediator free to diffuse in solution, immobilized in a thick, hydrated film, or adsorbed as a sub-monolayer on the electrode. The experiments can be performed with the goal to study the protein or to use it. Here emphasis is on mechanistic studies, which are easier in the configuration where the protein is adsorbed and electron transfer is direct, but we also explain the interpretation of signals obtained when diffusion processes affect the response.This chapter is organized as a series of responses to questions. Questions 1-5 are related to the basics of electrochemistry: what does "potential" or "current" mean, what does an electrochemical set-up look like? Questions 6-9 are related to the distinction between adsorbed and diffusive redox species. The answers to questions 10-13 explain the interpretation of slow and fast scan voltammetry with redox proteins. Questions 14-19 deal with catalytic electrochemistry, when the protein studied is actually an enzyme. Questions 20, 21 and 22 are general.

  5. Existence of aa3-type ubiquinol oxidase as a terminal oxidase in sulfite oxidation of Acidithiobacillus thiooxidans. (United States)

    Sugio, Tsuyoshi; Hisazumi, Tomohiro; Kanao, Tadayoshi; Kamimura, Kazuo; Takeuchi, Fumiaki; Negishi, Atsunori


    It was found that Acidithiobacillus thiooxidans has sulfite:ubiquinone oxidoreductase and ubiquinol oxidase activities in the cells. Ubiquinol oxidase was purified from plasma membranes of strain NB1-3 in a nearly homogeneous state. A purified enzyme showed absorption peaks at 419 and 595 nm in the oxidized form and at 442 and 605 nm in the reduced form. Pyridine ferrohaemochrome prepared from the enzyme showed an alpha-peak characteristic of haem a at 587 nm, indicating that the enzyme contains haem a as a component. The CO difference spectrum of ubiquinol oxidase showed two peaks at 428 nm and 595 nm, and a trough at 446 nm, suggesting the existence of an aa(3)-type cytochrome in the enzyme. Ubiquinol oxidase was composed of three subunits with apparent molecular masses of 57 kDa, 34 kDa, and 23 kDa. The optimum pH and temperature for ubiquinol oxidation were pH 6.0 and 30 degrees C. The activity was completely inhibited by sodium cyanide at 1.0 mM. In contrast, the activity was inhibited weakly by antimycin A(1) and myxothiazol, which are inhibitors of mitochondrial bc(1) complex. Quinone analog 2-heptyl-4-hydoroxyquinoline N-oxide (HOQNO) strongly inhibited ubiquinol oxidase activity. Nickel and tungstate (0.1 mM), which are used as a bacteriostatic agent for A. thiooxidans-dependent concrete corrosion, inhibited ubiquinol oxidase activity 100 and 70% respectively.

  6. Current status of NADPH oxidase research in cardiovascular pharmacology

    Directory of Open Access Journals (Sweden)

    Rodiño-Janeiro BK


    Full Text Available Bruno K Rodiño-Janeiro,1,2 Beatriz Paradela-Dobarro,1 María Isabel Castiñeiras-Landeira,1 Sergio Raposeiras-Roubín,1,3 José R González-Juanatey,1,3,4 Ezequiel Álvarez1,4 1Health Research Institute of Santiago de Compostela, Santiago de Compostela, Spain; 2European Molecular Biology Laboratory, Grenoble, France; 3Cardiology Department, University Clinic Hospital of Santiago de Compostela, Santiago de Compostela, Spain; 4Medicine Department, University of Santiago de Compostela, Santiago de Compostela, Spain Abstract: The implications of reactive oxygen species in cardiovascular disease have been known for some decades. Rationally, therapeutic antioxidant strategies combating oxidative stress have been developed, but the results of clinical trials have not been as good as expected. Therefore, to move forward in the design of new therapeutic strategies for cardiovascular disease based on prevention of production of reactive oxygen species, steps must be taken on two fronts, ie, comprehension of reduction-oxidation signaling pathways and the pathophysiologic roles of reactive oxygen species, and development of new, less toxic, and more selective nicotinamide adenine dinucleotide phosphate (NADPH oxidase inhibitors, to clarify both the role of each NADPH oxidase isoform and their utility in clinical practice. In this review, we analyze the value of NADPH oxidase as a therapeutic target for cardiovascular disease and the old and new pharmacologic agents or strategies to prevent NADPH oxidase activity. Some inhibitors and different direct or indirect approaches are available. Regarding direct NADPH oxidase inhibition, the specificity of NADPH oxidase is the focus of current investigations, whereas the chemical structure-activity relationship studies of known inhibitors have provided pharmacophore models with which to search for new molecules. From a general point of view, small-molecule inhibitors are preferred because of their hydrosolubility

  7. Pentamines as substrate for human spermine oxidase (United States)

    Takao, Koichi; Shirahata, Akira; Samejima, Keijiro; Casero, Robert A.; Igarashi, Kazuei; Sugita, Yoshiaki


    Substrate activities of various linear polyamines to human spermine oxidase (hSMO) were investigated. The activities were evaluated by monitoring the amount of H2O2 released from sample polyamines by hSMO. H2O2 was measured by a HPLC method that analyzed fluorescent dimers derived from the oxidation of homovanillic acid in the presence of horseradish peroxidase. Six triamines were tested and were found not to be hSMO substrates. Of sixteen tetramines tested, spermine (Spm) was the most active substrate, followed by homospermine and N-butylated Spm. Pentamines showed a characteristic pattern of substrate activity. Of thirteen pentamines tested, 3343 showed higher substrate activity than Spm, and 4343 showed similar activity to Spm. The activities of the other pentamines were as follows: 3443, 4443, 4344, 3344, 4334, 4444, and 3334 (in decreasing order). Product amines released from these pentamines by hSMO were then analyzed by HPLC. Triamine was the only observed product, and the amount of triamine was nearly equivalent to that of released H2O2. A marked difference in the pH dependency curves between tetramines and pentamines suggested that hSMO favored reactions with a non-protonated secondary nitrogen at the cleavage site. The Km and Vmax values for Spm and 3343 at pH 7.0 and 9.0 were consistent with the higher substrate activity of 3343 compared to Spm, as well as with the concept of a non-protonated secondary nitrogen at the cleavage site being preferred, and 3343 was well degraded at a physiological pH by hSMO. PMID:23449327

  8. Forage Polyphenol Oxidase and Ruminant Livestock Nutrition

    Directory of Open Access Journals (Sweden)

    Michael Richard F. Lee


    Full Text Available Polyphenol oxidase (PPO is associated with the detrimental effect of browning fruit and vegetables, however interest within PPO containing forage crops has grown since the brownng reaction was associated with reduced nitrogen (N losses in silo and the rumen. The reduction in protein breakdown in silo of red clover (high PPO forage increased the quality of protein, improving N-use efficiency (NUE when fed to ruminants. A further benefit of red clover silage feeding is a significant reduction in lipolysis in silo and an increase in the deposition of beneficial C18 polyunsaturated fatty acid (PUFA in animal products, which has also been linked to PPO activity. PPOs protection of plant protein and glycerol based-PUFA in silo is related to the deactivation of plant proteases and lipases. This deactivation occurs through PPO catalysing the conversion of diphenols to quinones which bind with cellular nucleophiles such as protein reforming a protein-bound phenol (PBP. If the protein is an enzyme the complexing denatures the enzyme. However, PPO is inactive in the anaerobic rumen and therefore any subsequent protection of plant protein and glycerol based-PUFA in the rumen must be as a result of events that occurred to the forage pre-ingestion. Reduced activity of plant proteases and lipases would have little effect on NUE and glycerol based-PUFA in the rumen due to the greater concentration of rumen microbial proteases and lipases. The mechanism for PPOs protection of plant protein in the rumen is a consequence of complexing plant protein, rather than protease deactivation per se. These complexed proteins reduce protein digestibility in the rumen and subsequently increase un-degraded dietary protein flow to the small intestine. The mechanism for protecting glycerol-based PUFA has yet to be fully elucidated but may be associated with entrapment within PBP reducing access to microbial lipases or differences in rumen digestion kinetics of red clover.

  9. Cytochemical localization of catalase and several hydrogen peroxide-producing oxidases in the nucleoids and matrix of rat liver peroxisomes

    NARCIS (Netherlands)

    Veenhuis, M.; Wendelaar Bonga, S.E.


    The distribution of catalase, amino acid oxidase, α-hydroxy acid oxidase, urate oxidase and alcohol oxidase was studied cytochemically in rat hepatocytes. The presence of catalase was demonstrated with the conventional diaminobenzidine technique. Oxidase activities were visualized with methods based

  10. Xanthine dehydrogenase and aldehyde oxidase impact plant hormone homeostasis and affect fruit size in 'Hass' avocado. (United States)

    Taylor, Nicky J; Cowan, A Keith


    The contribution of xanthine dehydrogenase (XDH, EC to fruit size was investigated using the normal and small-fruit variants of Persea americana Mill. cv. 'Hass'. Inhibition of XDH by treatment of normal fruit, in the linear phase of growth (phase II), with allopurinol (Allo) arrested fruit growth. Adenine (Ade), a less effective inhibitor of this enzyme, also arrested fruit growth when applied in phase II and slowed fruit growth when applied in phase III. A time-course study on the activity of XDH in mesocarp tissue from normal and small fruit showed that maximum activity occurred late in phase II and that the peak in activity was absent in mesocarp of the small fruit. Feeding Ade to growing fruit in phase III caused a transient decline in fruit growth (measured as change in fruit length). Thereafter, growth resumed although fruit size was irreversibly affected. Treatment of fruit with Ade and Ade-containing cytokinins altered activity of another molybdenum enzyme, aldehyde oxidase (EC Cytokinin oxidase was induced by cytokinin and auxin. Purine catabolism via hypoxanthine/xanthine was operative in normal fruit and in mesocarp from the small-fruit variant and as expected, Allo treatment caused accumulation of xanthine and adenine. In the absence of an increase in XDH during growth of the small-fruit phenotype, low levels of Ade were interpreted as resulting from respiration-enhanced adenylate depletion. Stress and/or pathogen induction of the alternative oxidase pathway is proposed as a possible cause.

  11. Mechanistic and stereochemical studies of glycine oxidase from Bacillus subtilis strain R5. (United States)

    Jamil, Farrukh; Gardner, Qurra-Tul-Ann Afza; Bashir, Qamar; Rashid, Naeem; Akhtar, Muhammad


    Glycine oxidase gene from a strain of Bacillus subtilis was cloned and expressed in Escherichia coli. The purified enzyme was found, by mass spectrometry, to have a protein M(r) of 40763 (value of 40761.6 predicted from DNA sequence) and a FAD prosthetic group M(r) of 785.1 (theoretical value of 785.5). Glycine oxidase optimally catalyzes the conversion of glycine and oxygen into glyoxylate, hydrogen peroxide, and ammonia. Using samples of [2-RS-(3)H(2),2-(14)C]-, [2-R-(3)H,2-(14)C]-, and [2-S-(3)H,2-(14)C]glycine, we found that in the overall process H(Si) is removed. Incubation of the enzyme with [2-RS-(3)H(2),2-(14)C]glycine under anaerobic conditions, when only the reducing half of the reaction can occur, led to the recovery of 98.5% of the original glycine, which had the same (3)H:(14)C ratio as the starting substrate. The primary isotope effect was studied using [2-(2)H(2)]glycine, and we found that the specificity constants, k(cat)/K(M), for the protio and deuterio substrates were 1.46 x 10(3) and 1.05 x 10(2) M(-1) s(-1), respectively. Two alternative mechanisms for FAD-containing oxidases that involve either the intermediacy of a FADH(2)-imino acid complex or an amino acid covalently linked to FAD, formed via a carbanion, have been considered. The current knowledge of the mechanisms is reviewed, and we argue that a mechanism involving the FADH(2)-imino acid complex can be dissected to satisfactorily explain some of puzzling observations for which the carbanion mechanism was originally conceived. Furthermore, our results, together with observations in the literature, suggest that the interaction of glycine with the enzyme occurs within a tight ternary complex, which is protected from the protons of the medium.

  12. Foreign language didactics: Identity questions

    Directory of Open Access Journals (Sweden)

    Michaela Píšová


    Full Text Available The efforts to emancipate foreign language didactics have been an ongoingand non-linear process launched in our country in the 50s of the previous century.Its key aspects may be presented as foreign language didactics identity questions.These include explicit delineation of the object and methodology of foreign languagedidactics on the basis of developmental analysis, current state of knowledge andtrends/approaches to the discipline. The issues related to the full-fledged scientificstatus of foreign language didactics are discussed both on a domain-general level(questions relevant for the whole field of subject didactics and on a domain-specificlevel (foreign language didactics specific questions.

  13. Dinuclear copper complexes with imidazole derivative ligands: a theoretical study related to catechol oxidase activity. (United States)

    Martínez, Ana; Membrillo, Ingrid; Ugalde-Saldívar, Victor M; Gasque, Laura


    Catechol oxidase is a very important and interesting metalloprotein. In spite of the efforts to understand the reaction mechanism of this protein, there are important questions that remain unanswered concerning the catalytic mechanism of this enzyme. In this article, dinuclear copper compounds are used as biomimetic models of catechol oxidase to study plausible reaction paths. These dinuclear copper(II) complexes have distant metal centers (of 7.5 Å approximately) and superior catalytic activity to that of many dicopper complexes with shorter Cu-Cu distances. One mononuclear copper(II) complex is also analyzed in this investigation in order to see the influence of the two metal centers in the catalytic activity. Density functional theory calculations were performed to obtain optimized structures, vertical ionization energies, vertical electron affinities, the electrodonating power (ω(-)), the electroaccepting power (ω(+)) and the energy difference of several reaction paths. The K(M) experimental results that were previously reported compare well with the electroaccepting power (ω(+)) of the copper compounds that are included in this article, indicating that this index is useful for the interpretation of the electron transfer capacity and therefore the catalytic activity. The catechol moiety coordinates to only one Cu ion, but two metal atoms are needed in order to have a good electron acceptor capacity of the biomimetic models.

  14. Alternative Energies

    Energy Technology Data Exchange (ETDEWEB)

    Planting, A.; De saint Jacob, Y.; Verwijs, H.; Belin, H.; Preesman, L.


    In two articles, one interview and one column attention is paid to alternative energies. The article 'A new light on saving energy' discusses the option to save energy by modernising lighting systems in urban areas. The column 'View from Paris' focuses on investment decisions in France with regard to renewable energy and energy savings. The article 'Europe turns a blind eye to big battery' discusses developments in batteries to store energy. The interview concerns fuel cell expert and formerly President of UTC Power Jan van Dokkum. The last article gives a brief overview of the European Energy Research Alliance (EERA) and the challenges this alliance will have to face with regard to climate change and energy security.

  15. Regulation of NADPH oxidase activity in phagocytes: relationship between FAD/NADPH binding and oxidase complex assembly. (United States)

    Debeurme, Franck; Picciocchi, Antoine; Dagher, Marie-Claire; Grunwald, Didier; Beaumel, Sylvain; Fieschi, Franck; Stasia, Marie-José


    The X(+)-linked chronic granulomatous disease (X(+)-CGD) variants are natural mutants characterized by defective NADPH oxidase activity but with normal Nox2 expression. According to the three-dimensional model of the cytosolic Nox2 domain, most of the X(+)-CGD mutations are located in/or close to the FAD/NADPH binding regions. A structure/function study of this domain was conducted in X(+)-CGD PLB-985 cells exactly mimicking 10 human variants: T341K, C369R, G408E, G408R, P415H, P415L, Δ507QKT509-HIWAinsert, C537R, L546P, and E568K. Diaphorase activity is defective in all these mutants. NADPH oxidase assembly is normal for P415H/P415L and T341K mutants where mutation occurs in the consensus sequences of NADPH- and FAD-binding sites, respectively. This is in accordance with their buried position in the three-dimensional model of the cytosolic Nox2 domain. FAD incorporation is abolished only in the T341K mutant explaining its absence of diaphorase activity. This demonstrates that NADPH oxidase assembly can occur without FAD incorporation. In addition, a defect of NADPH binding is a plausible explanation for the diaphorase activity inhibition in the P415H, P415L, and C537R mutants. In contrast, Cys-369, Gly-408, Leu-546, and Glu-568 are essential for NADPH oxidase complex assembly. However, according to their position in the three-dimensional model of the cytosolic domain of Nox2, only Cys-369 could be in direct contact with cytosolic factors during oxidase assembly. In addition, the defect in oxidase assembly observed in the C369R, G408E, G408R, and E568K mutants correlates with the lack of FAD incorporation. Thus, the NADPH oxidase assembly process and FAD incorporation are closely related events essential for the diaphorase activity of Nox2.

  16. Some properties of active and latent catechol oxidase of mushroom

    Directory of Open Access Journals (Sweden)

    Janusz Czapski


    Full Text Available Latent form of mushroom catechol oxidase was activated by O,1% sodium dodecyl sulfate (SDS. Catalytic power of the latent form, calculated from the kinetic parameters was 1,8 times higher than that of active one. Salicyl hydroxamic acid (SHAM appeared as a powerful inhibitor for both active and latent forms of catechol oxidase. However, in the range of 150-250 μM SHAM the inhibitory effect for active catechol oxidase was significantly higher than that for the latent one. Non-competitive and irreversible characteristics of inhibition of latent and active catechol oxidase was calculated from kinetic data. Electrophoretic analysis followed by scanning of the gels was used. The spots' absorbance was determined from a computer image of the isoenzyme band patterns. It allowed us to estimate gels quantitatively. Presence of one additional clearly defined slow moving isoform of SDS-activated catechol oxidase, differed in the respect of 3 bands for the active and 4 bands for the total.

  17. Calcium transport in vesicles energized by cytochrome oxidase

    Energy Technology Data Exchange (ETDEWEB)

    Rosier, Randy N. [Univ. of Rochester, NY (United States)


    Experiments on the reconstitution of cytochrome oxidase into phospholipid vesicles were carried out using techniques of selectivity energizing the suspensions with ascorbate and cytochrome c or ascorbate, PMS, and internally trapped cytochrome c. It was found that the K+ selective ionophore valinomycin stimulated the rate of respiration of cytochrome oxidase vesicles regardless of the direction of the K+ flux across the vesicle membranes. The stimulation occurred in the presence of protonophoric uncouplers and in the complete absence of potassium or in detergent-lysed suspensions. Gramicidin had similar effects and it was determined that the ionophores acted by specific interaction with cytochrome oxidase rather than by the previously assumed collapse of membrane potentials. When hydrophobic proteins and appropriate coupling factors were incorporated into the cytochrome oxidase, vesicles phosphorylation of ADP could be coupled to the oxidation reaction of cytochrome oxidase. Relatively low P:O, representing poor coupling of the system, were problematical and precluded measurements of protonmotive force. However the system was used to study ion translocation.

  18. Cation binding site of cytochrome c oxidase: progress report. (United States)

    Vygodina, Tatiana V; Kirichenko, Anna; Konstantinov, Alexander A


    Cytochrome c oxidase from bovine heart binds Ca(2+) reversibly at a specific Cation Binding Site located near the outer face of the mitochondrial membrane. Ca(2+) shifts the absorption spectrum of heme a, which allowed earlier the determination of the kinetic and equilibrium characteristics of the binding, and, as shown recently, the binding of calcium to the site inhibits cytochrome oxidase activity at low turnover rates of the enzyme [Vygodina, Т., Kirichenko, A., Konstantinov, A.A (2013). Direct Regulation of Cytochrome c Oxidase by Calcium Ions. PloS ONE 8, e74436]. This paper summarizes further progress in the studies of the Cation Binding Site in this group presenting the results to be reported at 18th EBEC Meeting in Lisbon, 2014. The paper revises specificity of the bovine oxidase Cation Binding Site for different cations, describes dependence of the Ca(2+)-induced inhibition on turnover rate of the enzyme and reports very high affinity binding of calcium with the "slow" form of cytochrome oxidase. This article is part of a Special Issue entitled: 18th European Bioenergetic Conference. Guest Editors: Manuela Pereira and Miguel Teixeira.

  19. Crystal Structure of Alcohol Oxidase from Pichia pastoris.

    Directory of Open Access Journals (Sweden)

    Christian Koch

    Full Text Available FAD-dependent alcohol oxidases (AOX are key enzymes of methylotrophic organisms that can utilize lower primary alcohols as sole source of carbon and energy. Here we report the crystal structure analysis of the methanol oxidase AOX1 from Pichia pastoris. The crystallographic phase problem was solved by means of Molecular Replacement in combination with initial structure rebuilding using Rosetta model completion and relaxation against an averaged electron density map. The subunit arrangement of the homo-octameric AOX1 differs from that of octameric vanillyl alcohol oxidase and other dimeric or tetrameric alcohol oxidases, due to the insertion of two large protruding loop regions and an additional C-terminal extension in AOX1. In comparison to other alcohol oxidases, the active site cavity of AOX1 is significantly reduced in size, which could explain the observed preference for methanol as substrate. All AOX1 subunits of the structure reported here harbor a modified flavin adenine dinucleotide, which contains an arabityl chain instead of a ribityl chain attached to the isoalloxazine ring.

  20. Questions of Modern Cosmology Galileo's Legacy

    CERN Document Server

    D'Onofrio, Mauro


    Are we living in the "golden age" of cosmology? Are we close to understanding the nature of the unknown ingredients of the currently most accepted cosmological model and the physics of the early Universe? Or are we instead approaching a paradigm shift? What is dark matter and does it exist? How is it distributed around galaxies and clusters? Is the scientific community open to alternative ideas that may prompt a new scientific revolution - as the Copernican revolution did in Galileo's time? Do other types of supernovae exist that can be of interest for cosmology? Why have quasars never been effectively used as standard candles? Can you tell us about the scientific adventure of COBE? How does the extraction of the Cosmic Microwave Background anisotropy depend on the subtraction of the various astrophysical foregrounds? These, among many others, are the astrophysical, philosophical and sociological questions surrounding modern cosmology and the scientific community that Mauro D'Onofrio and Carlo Burigana pose t...

  1. Pyruvate oxidase influences the sugar utilization pattern and capsule production in Streptococcus pneumoniae.

    Directory of Open Access Journals (Sweden)

    Sandra M Carvalho

    Full Text Available Pyruvate oxidase is a key function in the metabolism and lifestyle of many lactic acid bacteria and its activity depends on the presence of environmental oxygen. In Streptococcus pneumoniae the protein has been suggested to play a major role in metabolism and has been implicated in virulence, oxidative stress survival and death in stationary phase. Under semi-aerobic conditions, transcriptomic and metabolite profiling analysis of a spxB mutant grown on glucose showed minor changes compared to the wild type, apart from the significant induction of two operons involved in carbohydrate uptake and processing. This induction leads to a change in the sugar utilization capabilities of the bacterium, as indicated by the analysis of the growth profiles of the D39 parent and spxB mutant on alternative carbohydrates. Metabolic analysis and growth experiments showed that inactivation of SpxB has no effect on the glucose fermentation pattern, except under aerobic conditions. More importantly, we show that mutation of spxB results in the production of increased amounts of capsule, the major virulence factor of S. pneumoniae. Part of this increase can be attributed to induction of capsule operon (cps transcription. Therefore, we propose that S. pneumoniae utilizes pyruvate oxidase as an indirect sensor of the oxygenation of the environment, resulting in the adaption of its nutritional capability and the amount of capsule to survive in the host.

  2. $^{111m}$Cd- and $^{199m}$Hg-derivatives of blue oxidases

    CERN Multimedia


    The rack-induced bonding concept (H.B.Gray & B.G.~Malmstroem, Comments Inorg. Chem, 2, 203, 1983) postulates that the bound metal ion in metalloproteins is forced to adopt a coordination geometry determined by the rigid peptide conformation of the protein. Alternatively, the metal ion could create its own favoured coordination geometry in a soft peptide conformation. In order to decide who is slave or master the changes of coordination and rigidity of metal sites in blue copper proteins due to metal and ligand exchange were studied by $^{111m}$Cd and $^{199m}$Hg $\\gamma$-$\\gamma$-perturbed angular correlation (PAC). To get a better understanding of the so called " Type 1 Copper Site " of the blue oxidases laccase (LAC) and ascorbate oxidase (AO) we concentrated our investigations on the small blue copper proteins azurin and plastocyanin. \\\\ \\\\In azurin~(Az), the metal ligand methionine 121~(M121) was replaced by several amino acids, e.g. asparagine~(N), glutamic acid~(E), via site directed mutagenesis. Di...

  3. Urate Oxidase Knockdown Decreases Oxidative Stress in a Murine Hepatic Cell Line

    Directory of Open Access Journals (Sweden)

    Beth M. Cleveland


    Full Text Available Humans, birds, and some primates do not express the uric acid degrading enzyme urate oxidase (UOX and, as a result, have plasma uric acid concentrations higher than UOX expressing animals. Although high uric acid concentrations are suggested to increase the antioxidant defense system and provide a health advantage to animals without UOX, knockout mice lacking UOX develop pathological complications including gout and kidney failure. As an alternative to the knockout model, RNA interference was used to decrease UOX expression using stable transfection in a mouse hepatic cell line (ATCC, FL83B. Urate oxidase mRNA was reduced 66% (p < 0.05 compared to wild type, as measured by real time RT-PCR. To determine if UOX knockdown resulted in enhanced protection against oxidative stress, cells were challenged with hexavalent chromium (Cr(VI or 3-morpholinosydnonimine hydrochloride (SIN-1. Compared to wild type, cells with UOX knockdown exhibited a 37.2 ± 3.5% reduction (p < 0.05 in the electron spin resonance (ESR signal after being exposed to Cr(VI and displayed less DNA fragmentation (p < 0.05 following SIN-1 treatment. Cell viability decreased in wild type cells (p < 0.05, but not cells with UOX knockdown, after treatment with SIN-1. These results are consistent with an increased intracellular uric acid concentration and an increased defense against oxidative stress.

  4. Automated cytochrome c oxidase bioassay developed for ionic liquids' toxicity assessment. (United States)

    Costa, Susana P F; Martins, Bárbara S F; Pinto, Paula C A G; Saraiva, M Lúcia M F S


    A fully automated cytochrome c oxidase assay resorting to sequential injection analysis (SIA) was developed for the first time and implemented to evaluate potential toxic compounds. The bioassay was validated by evaluation of 15 ionic liquids (ILs) with distinct cationic head groups, alkyl side chains and anions. The assay was based on cytochrome c oxidase activity reduction in presence of tested compounds and quantification of inhibitor concentration required to cause 50% of enzyme activity inhibition (EC50). The obtained results demonstrated that enzyme activity was considerably inhibited by BF4 anion and ILs incorporating non-aromatic pyrrolidinium and tetrabutylphosphonium cation cores. Emim [Ac] and chol [Ac], on contrary, presented the higher EC50 values among the ILs tested. The developed automated SIA methodology is a simple and robust high-throughput screening bioassay and exhibited good repeatability in all the tested conditions (rsd<3.7%, n=10). Therefore, it is expected that due to its simplicity and low cost, the developed approach can be used as alternative to traditional screening assays for evaluation of ILs toxicity and identification of possible toxicophore structures. Additionally, the results presented in this study provide further information about ILs toxicity.

  5. Arts and Techniques of Questioning

    Institute of Scientific and Technical Information of China (English)

    Sun Yu


    Most learners learn English from teachers in classroom. Therefore, classroom instruction plays a very important role. Teachers' questioning constitutes a very significant aspect of classroom teaching. It is not only an important part of classroom interaction but an effective way of learning second language as well.

  6. Understanding Bitcoins: Facts and Questions


    Bruno Saboia de Albuquerque; Marcelo de Castro Callado


    The objective of this work is to do a research challenge about the digital currency named Bitcoins, as well as exploit the general concept behind digital currencies and cryptocurrencies, and enumerate some of its current criticism and problems. Such currencies usage and public knowledge is increasing hastily on the last few months, and many questions arise with its popularity.

  7. Questioning Mechanisms During Complex Learning (United States)


    is probably a general phenomenon in this culture that tutors are reluctant to give negative feedback on students’ errors and poor answers. Instead of...1991). Questioning in classrooms: A sociolinguistic perspective. Review of £’fiu!r’aiona1 R.ear.h, L, 157-178. Carroll, J. M., Mack, R. L., Lewis, C

  8. Understanding Bitcoins: Facts and Questions

    Directory of Open Access Journals (Sweden)

    Bruno Saboia de Albuquerque


    Full Text Available The objective of this work is to do a research challenge about the digital currency named Bitcoins, as well as exploit the general concept behind digital currencies and cryptocurrencies, and enumerate some of its current criticism and problems. Such currencies usage and public knowledge is increasing hastily on the last few months, and many questions arise with its popularity.

  9. Forage polyphenol oxidase and ruminant livestock nutrition. (United States)

    Lee, Michael R F


    Polyphenol oxidase (PPO) is predominately associated with the detrimental effect of browning fruit and vegetables, however, interest within PPO containing forage crops (crops to be fed to animals) has grown since the browning reaction was associated with reduced nitrogen (N) losses in silo and the rumen. The reduction in protein breakdown in silo of red clover (high PPO forage) increased the quality of protein, improving N-use efficiency [feed N into product N (e.g., Milk): NUE] when fed to ruminants. A further benefit of red clover silage feeding is a significant reduction in lipolysis (cleaving of glycerol-based lipid) in silo and an increase in the deposition of beneficial C18 polyunsaturated fatty acid (PUFA) in animal products, which has also been linked to PPO activity. PPOs protection of plant protein and glycerol based-PUFA in silo is related to the deactivation of plant proteases and lipases. This deactivation occurs through PPO catalyzing the conversion of diphenols to quinones which bind with cellular nucleophiles such as protein reforming a protein-bound phenol (PBP). If the protein is an enzyme (e.g., protease or lipase) the complexing denatures the enzyme. However, PPO is inactive in the anaerobic rumen and therefore any subsequent protection of plant protein and glycerol based-PUFA in the rumen must be as a result of events that occurred to the forage pre-ingestion. Reduced activity of plant proteases and lipases would have little effect on NUE and glycerol based-PUFA in the rumen due to the greater concentration of rumen microbial proteases and lipases. The mechanism for PPOs protection of plant protein in the rumen is a consequence of complexing plant protein, rather than protease deactivation per se. These complexed proteins reduce protein digestibility in the rumen and subsequently increase undegraded dietary protein flow to the small intestine. The mechanism for protecting glycerol-based PUFA has yet to be fully elucidated but may be associated

  10. Alternate Realities (United States)

    Jones, Robert


    Two identical learners, observing different example input, or the same examples, but in different order, can form different categories and so judge newer/later input differently. (Machine Learning, T. Mitchell, McGraw Hill, 1997 and Asa H., R. Jones, Trans. Kansas Acad. Sci., vol 109, # 3/4, pg 159, 2006) It seems certain that each of us experiences a somewhat different reality, the question is just how widely these realities can vary one from another. Perhaps 4% of people exhibit synesthesia, perceiving letters or numbers as colored, numbers and dates as having personalities or occupying locations in space. (Synesthesia, R. Cytowic, MIT Press, 2002) The Sapir- Whorf hypothesis claims that a speakers language influences his category structure and the way he thinks. (Language, thought, and reality, B. Whorf, MIT Press, 1956) Those who are skillful at mathematics may know an additional language and be able to think thoughts that the layman can not. The philosophers Plato and Descartes claimed to have had, at certain moments in their lives, a new view of the world, its basic constituents, and its rules which were totally different from our conventional view of reality. (Reflections on Kurt Godel, H. Wang, MIT Press, 1987, pg. 46) Fairly large scale differences are experienced by those who believe in (make use of) concepts like spirit(s), soul(s), god(s), life after death, platonism or Everett's many worlds interpretation of quantum mechanics (The Physics of Immortality, F. Tipler, Doubleday, 1994, pg. 176) )

  11. Fluorescent Probes for Analysis and Imaging of Monoamine Oxidase Activity

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Dokyoung; Jun, Yong Woong; Ahn, Kyo Han [POSTECH, Pohang (Korea, Republic of)


    Monoamine oxidases catalyze the oxidative deamination of dietary amines and amine neurotransmitters, and assist in maintaining the homeostasis of the amine neurotransmitters in the brain. Dysfunctions of these enzymes can cause neurological and behavioral disorders including Parkinson's and Alzheimer's diseases. To understand their physiological roles, efficient assay methods for monoamine oxidases are essential. Reviewed in this Perspective are the recent progress in the development of fluorescent probes for monoamine oxidases and their applications to enzyme assays in cells and tissues. It is evident that still there is strong need for a fluorescent probe with desirable substrate selectivity and photophysical properties to challenge the much unsolved issues associated with the enzymes and the diseases.

  12. Characterization of wheat germin (oxalate oxidase) expressed by Pichia pastoris (United States)

    Pan, Heng-Yen; Whittaker, Mei M.; Bouveret, Romaric; Berna, Anne; Bernier, François; Whittaker, James W.


    High-level secretory expression of wheat (Triticum aestivum) germin/oxalate oxidase was achieved in Pichia pastoris fermentation cultures as an α-mating factor signal peptide fusion, based on the native wheat cDNA coding sequence. The oxalate oxidase activity of the recombinant enzyme is substantially increased (7-fold) by treatment with sodium periodate, followed by ascorbate reduction. Using these methods, approximately 1 g (4×104 U) of purified, activated enzyme was obtained following eight days of induction of a high density Pichia fermentation culture, demonstrating suitability for large-scale production of oxalate oxidase for biotechnological applications. Characterization of the recombinant protein shows that it is glycosylated, with N-linked glycan attached at Asn47. For potential biomedical applications, a nonglycosylated (S49A) variant was also prepared which retains essentially full enzyme activity, but exhibits altered protein-protein interactions. PMID:17399681

  13. A mammalian H+ channel generated through alternative splicing of the NADPH oxidase homolog NOH-1

    NARCIS (Netherlands)

    Bánfi, B; Maturana, A; Jaconi, S; Arnaudeau, S; Laforge, T; Sinha, B; Ligeti, E; Demaurex, N; Krause, K H


    Voltage-gated proton (H+) channels are found in many human and animal tissues and play an important role in cellular defense against acidic stress. However, a molecular identification of these unique ion conductances has so far not been achieved. A 191-amino acid protein is described that, upon hete

  14. Analysis of variance: Comfortless questions


    L.V. Nedorezov


    In this paper the simplest variant of analysis of variance is under consideration. Three examples from textbooks by Lakin (1990) and Rokitsky (1973) were re-considered. It was obtained that traditional one-way ANOVA and Kruskal - Wallis criterion can lead to unreal results about factor's influence on value of characteristics. Alternative way to solution of the same problem is under consideration too.

  15. In vitro xanthine oxidase inhibitory activity of leaves, fruits and peel extracts of Citrus aurantium, Citrus limetta and Citrus limon

    Directory of Open Access Journals (Sweden)

    Muthiah PL


    Full Text Available Aim of the study: To evaluate the in vitro xanthine oxidase inhibitory activity of the extract of leaves, fruits and peel of Citrus aurantium, Citrus limetta and Citrus limon.   Materials and Methods: Xanthine oxidase inhibitory activity was assayed spectrophotometrically under aerobic conditions and the degree of enzyme inhibition was determined by measuring the increase in absorbance at 295nm associated with uric acid formation.   Results: Among the extracts tested, the C.limetta peel extract exhibited highest potency of xanthine oxidase inhibition (IC50 40.16±0.88μg/ml. This was followed by C.aurantium peel (IC50 51.50±2.05μg/ml, C.limon peel (IC50 64.90±1.24μg/ml, C.aurantium leaf (IC5073.50±1.26μg/ml, C.limetta leaf (IC50 74.83±2.42μg/ml, C.limon leaf (IC50 76.83±2.02μg/ml, C.limetta fruit (IC50 95.16±0.60μg/ml extracts compared with the IC50 value of standard allopurinol was 6.6μg/ml.   Conclusion: Recent findings show that the occurrence of gout is increasing worldwide, possibly due to the changes in dietary habits like intake of food rich in nucleic acids, such as meat, sea foods, etc. Xanthine oxidase inhibitors such as allopurinol is the drug of choice, however it has been observed more side effects.  An alternative to allopurinol is the use of medicinal plants, We thus began our program to look for xanthine oxidase inhibitors of phytochemical origin. In conclusion, the study suggests that the leaves and peel extracts of Citrus aurantium, Citrus limetta and Citrus limon possess xanthine oxidase inhibitory activity that might be helpful in preventing or slowing the progress of gout and related disorders.

  16. Multi-function oxidases are responsible for the synergistic interactions occurring between repellents and insecticides in mosquitoes

    Directory of Open Access Journals (Sweden)

    Duchon Stéphane


    Full Text Available Abstract Background With the spread of pyrethroid resistance in mosquitoes, the combination of an insecticide (carbamate or organophosphate with a repellent (DEET is considered as a promising alternative strategy for the treatment of mosquito nets and other relevant materials. The efficacy of these mixtures comes from the fact that they reproduce pyrethroid features and that positive interactions occur between insecticides and repellent. To better understand the mechanisms involved and assess the impact of detoxifying enzymes (oxidases and esterases in these interactions, bioassays were carried out in the laboratory against the main dengue vector Aedes aegypti. Methods Topical applications of DEET and propoxur (carbamate, used alone or as a mixture, were carried out on female mosquitoes, using inhibitors of the two main detoxification pathways in the insect. PBO, an inhibitor of multi-function oxidases, and DEF, an inhibitor of esterases, were applied one hour prior to the main treatment. Results Results showed that synergism between DEET and propoxur disappeared in the presence of PBO but not with DEF. This suggests that oxidases, contrary to esterases, play a key role in the interactions occurring between DEET and cholinesterase inhibitors in mosquitoes. Conclusion These findings are of great interest for the implementation of "combination nets" in the field. They support the need to combine insecticide with repellent to overcome insecticide resistance in mosquitoes of public health importance.

  17. Gold-Coated M13 Bacteriophage as a Template for Glucose Oxidase Biofuel Cells with Direct Electron Transfer. (United States)

    Blaik, Rita A; Lan, Esther; Huang, Yu; Dunn, Bruce


    Glucose oxidase-based biofuel cells are a promising source of alternative energy for small device applications, but still face the challenge of achieving robust electrical contact between the redox enzymes and the current collector. This paper reports on the design of an electrode consisting of glucose oxidase covalently attached to gold nanoparticles that are assembled onto a genetically engineered M13 bacteriophage using EDC-NHS chemistry. The engineered phage is modified at the pIII protein to attach onto a gold substrate and serves as a high-surface-area template. The resulting "nanomesh" architecture exhibits direct electron transfer (DET) and achieves a higher peak current per unit area of 1.2 mA/cm(2) compared to most other DET attachment schemes. The final enzyme surface coverage on the electrode was calculated to be approximately 4.74 × 10(-8) mol/cm(2), which is a significant improvement over most current glucose oxidase (GOx) DET attachment methods.

  18. Functional characterization of gibberellin oxidases from cucumber, Cucumis sativus L. (United States)

    Pimenta Lange, Maria João; Liebrandt, Anja; Arnold, Linda; Chmielewska, Sara-Miriam; Felsberger, André; Freier, Eduard; Heuer, Monika; Zur, Doreen; Lange, Theo


    Cucurbits have been used widely to elucidate gibberellin (GA) biosynthesis. With the recent availability of the genome sequence for the economically important cucurbit Cucumis sativus, sequence data became available for all genes potentially involved in GA biosynthesis for this species. Sixteen cDNAs were cloned from root and shoot of 3-d to 7-d old seedlings and from mature seeds of C. sativus. Two cDNAs code for GA 7-oxidases (CsGA7ox1, and -2), five for GA 20-oxidases (CsGA20ox1, -2, -3, -4, and -5), four for GA 3-oxidases (CsGA3ox1, -2, -3, and -4), and another five for GA 2-oxidases (CsGA2ox1, -2, -3, -4, and -5). Their enzymatic activities were investigated by heterologous expression of the cDNAs in Escherichia coli and incubation of the cell lysates with (14)C-labelled, D2-labelled, or unlabelled GA-substrates. The two GA 7-oxidases converted GA12-aldehyde to GA12 efficiently. CsGA7ox1 converted GA12 to GA14, to 15α-hydroxyGA12, and further to 15α-hydroxyGA14. CsGA7ox2 converted GA12 to its 12α-hydroxylated analogue GA111. All five GA 20-oxidases converted GA12 to GA9 as a major product, and to GA25 as a minor product. The four GA 3-oxidases oxidized the C19-GA GA9 to GA4 as the only product. In addition, three of them (CsGA3ox2, -3, and -4) converted the C20-GA GA12 to GA14. The GA 2-oxidases CsGA2ox1, -2, -3, and -4 oxidized the C19-GAs GA9 and GA4 to GA34 and GA51, respectively. CsGA2ox2, -3, and -4 converted GA51 and GA34 further to respective GA-catabolites. In addition to C19-GAs, CsGA2ox4 also converted the C20-GA GA12 to GA110. In contrast, CsGA2ox5 oxidized only the C20 GA12 to GA110 as the sole product. As shown for CsGA20ox1 and CsGA3ox1, similar reactions were catalysed with 13-hydroxlyated GAs as substrates. It is likely that these enzymes are also responsible for the biosynthesis of 13-hydroxylated GAs in vivo that occur at low levels in cucumber.

  19. A Conserved Steroid Binding Site in Cytochrome c Oxidase

    Energy Technology Data Exchange (ETDEWEB)

    Qin, Ling; Mills, Denise A.; Buhrow, Leann; Hiser, Carrie; Ferguson-Miller, Shelagh (Michigan)


    Micromolar concentrations of the bile salt deoxycholate are shown to rescue the activity of an inactive mutant, E101A, in the K proton pathway of Rhodobacter sphaeroides cytochrome c oxidase. A crystal structure of the wild-type enzyme reveals, as predicted, deoxycholate bound with its carboxyl group at the entrance of the K path. Since cholate is a known potent inhibitor of bovine oxidase and is seen in a similar position in the bovine structure, the crystallographically defined, conserved steroid binding site could reveal a regulatory site for steroids or structurally related molecules that act on the essential K proton path.

  20. Colloidal properties of biomacromolecular solutions: Towards urate oxidase crystal design (United States)

    Bonneté, Françoise


    Crystallization of biological macromolecules is governed by weak interaction forces, attractive and repulsive. Knowledge of solution properties, via second virial coefficient measurements, makes it possible to select physico-chemical parameters that govern and control phase diagrams and thus to grow crystals for specific applications (bio-crystallography or pharmaceutical processes). We highlight here with urate oxidase a salting-in effect that increases its solubility and the depletion effect of amphiphilic polymer, at a polymer concentration above its cmc, in order to grow diffracting crystals of urate oxidase. These two effects were used to grow crystals for high pressure crystallography and in a purification process.

  1. Development of new radiopharmaceuticals for imaging monoamine oxidase B

    Energy Technology Data Exchange (ETDEWEB)

    Vasdev, Neil, E-mail:; Sadovski, Oleg; Moran, Matthew D.; Parkes, Jun; Meyer, Jeffrey H.; Houle, Sylvain; Wilson, Alan A.


    Introduction: Imaging monoamine oxidase B (MAO-B) in the central nervous system with PET is an important goal for psychiatric studies. We here report an improved and automated radiosynthesis of N-(6-[{sup 18}F]-fluorohexyl)-N-methylpropargylamine ([{sup 18}F]FHMP; [{sup 18}F]-1), as well as the radiosynthesis of two new promising candidates for imaging cerebral MAO-B, namely, carbon-11-labeled 3-(4-[{sup 11}C]-methoxyphenyl)-6-methyl-2H-1-benzopyran-2-one ([{sup 11}C]-2) and N-((1H-pyrrol-2-yl)methyl)-N-[{sup 11}C]-methyl-1-phenylmethanamine ([{sup 11}C]-3). Methods: Fluorine-18-labeled 1 was prepared via a tosyloxy precursor in 29%{+-}5% uncorrected radiochemical yield, relative to [{sup 18}F]-fluoride. Both carbon-11-labeled compounds were prepared with [{sup 11}C]CH{sub 3}I using the 'LOOP' method in 11% and 18% uncorrected radiochemical yields, respectively, relative to starting [{sup 11}C]CO{sub 2}. All radiotracers had specific activities >37 GBq/{mu}mol and were >98% radiochemically pure at end of synthesis (<40 min). All radiotracers were evaluated by ex vivo biodistribution studies in conscious rodents. Results: A major radioactive metabolite in the rodent brain was observed following administration of [{sup 18}F]-1. While [{sup 11}C]-2 had moderate brain penetration and good clearance from normal brain tissue, distribution of radioactivity in brain was indicative of free and nonspecific binding. Good brain uptake was observed with [{sup 11}C]-3 (0.8%-1.4% injected dose per gram at 5 min postinjection), binding appeared to be reversible and distribution conformed with regional distribution of MAO-B in the rat brain. Preinjection of 3 or L-deprenyl showed a modest reduction (up to 25%) of brain activity. Conclusion: Carbon-11-labeled 3 was found to have the most favorable properties of the radiotracers evaluated; however, the signal-to-noise ratio was too low to warrant further in vivo imaging studies. Alternative radiotracers for imaging MAO

  2. Direct Enzymatic Assay for Alcohol Oxidase, Alcohol Dehydrogenase, and Formaldehyde Dehydrogenase in Colonies of Hansenula polymorpha


    Eggeling, L; Sahm, H


    A procedure is described for the qualitative direct identification of alcohol oxidase, alcohol dehydrogenase, and formaldehyde dehydrogenase in yeast colonies. The method has been applied successfully to isolate mutants of Hansenula polymorpha with altered glucose repression of alcohol oxidase.

  3. On a question of Gross (United States)

    Banerjee, Abhijit


    Using the notion of weighted sharing of sets we prove two uniqueness theorems which improve the results proved by Fang and Qiu [H. Qiu, M. Fang, A unicity theorem for meromorphic functions, Bull. Malaysian Math. Sci. Soc. 25 (2002) 31-38], Lahiri and Banerjee [I. Lahiri, A. Banerjee, Uniqueness of meromorphic functions with deficient poles, Kyungpook Math. J. 44 (2004) 575-584] and Yi and Lin [H.X. Yi, W.C. Lin, Uniqueness theorems concerning a question of Gross, Proc. Japan Acad. Ser. A 80 (2004) 136-140] and thus provide an answer to the question of Gross [F. Gross, Factorization of meromorphic functions and some open problems, in: Proc. Conf. Univ. Kentucky, Lexington, KY, 1976, in: Lecture Notes in Math., vol. 599, Springer, Berlin, 1977, pp. 51-69], under a weaker hypothesis.

  4. The study of a fluorescent biosensor based on polyelectrolyte microcapsules with encapsulated glucose oxidase (United States)

    Kazakova, L. I.; Sirota, N. P.; Sirota, T. V.; Shabarchina, L. I.


    A fluorescent biosensor is synthesized and described. The biosensor consists of polyelectrolyte microcapsules with glucose oxidase (GOx) entrapped in the cavities and an oxygen-sensitive fluorescent indicator Ru(dpp) immobilized in shells, where Ru(dpp) is tris(4,7-diphenyl-1,10-phenanthroline)ruthenium(II) dichloride. The theoretical activity of the encapsulated GOx and the effect storage time and medium composition have on the stability of sensor microcapsules are determined from polarographic measurements. No change in the activity of the encapsulated enzyme and or its loss to the storage medium are detected over the test period. The dispersion medium (water or a phosphate buffer) are shown to have no effect on the activity of microcapsules with immobilized GOx. The described optical sensor could be used as an alternative to electrochemical sensors for in vitro determination of glucose in the clinically important range of concentrations (up to 10 mmol/L).

  5. Characterization of water-forming NADH oxidases for co-factor regeneration

    DEFF Research Database (Denmark)

    Rehn, Gustav; Pedersen, Asbjørn Toftgaard; J. Charnock, Simon

    an environmentaland economic perspective [1]. Alcohol dehydrogenases (ADH) offer one such alternative. However, the reaction requires the oxidized nicotinamide co-factor (NAD+) that must be recycled due to its high cost contribution. One regeneration method that offers certain advantages is the oxidation of NADH...... using water forming NADH oxidases (NOX-2). The implementation of the ADH/NOX system for alcohol oxidation, however, requires consideration of several different issues. Enzyme activity and stability at relevant pH and temperature conditions, but also the tolerance to the substrates and products present......Traditional chemical methods for alcohol oxidation are often associated with issues such as high consumption of expensive oxidizing agents, generation of metal waste and the use of environmentally undesirable organic solvents. Developing green, selective catalysts is therefore important from...

  6. Foreign language didactics: Identity questions


    Michaela Píšová


    The efforts to emancipate foreign language didactics have been an ongoingand non-linear process launched in our country in the 50s of the previous century.Its key aspects may be presented as foreign language didactics identity questions.These include explicit delineation of the object and methodology of foreign languagedidactics on the basis of developmental analysis, current state of knowledge andtrends/approaches to the discipline. The issues related to the full-fledged scientificstatus of ...

  7. Some open questions in hydrodynamics

    CERN Document Server

    Dyndal, Mateusz


    When speaking of unsolved problems in physics, this is surprising at first glance to discuss the case of fluid mechanics. However, there are many deep open questions that come with the theory of fluid mechanics. In this paper, we discuss some of them that we classify in two categories, the long term behavior of solutions of equations of hydrodynamics and the definition of initial (boundary) conditions. The first set of questions come with the non-relativistic theory based on the Navier-Stokes equations. Starting from smooth initial conditions, the purpose is to understand if solutions of Navier-Stokes equations remain smooth with the time evolution. Existence for just a finite time would imply the evolution of finite time singularities, which would have a major influence on the development of turbulent phenomena. The second set of questions come with the relativistic theory of hydrodynamics. There is an accumulating evidence that this theory may be relevant for the description of the medium created in high en...

  8. Questions as a tool to design

    DEFF Research Database (Denmark)

    Aurisicchio, Marco; Ahmed, Saeema; Wallace, Ken


    This paper describes an interpretation of design activity through investigating design questions. From a number of previous studies two types of question have been identified: 1) reasoning questions; and 2) strategic questions. Strategic questions are part of an experienced designers approach to ...

  9. Soft Physics and Intermittency Open Question(s) in Krakow

    CERN Document Server

    Peschanski, R


    This contribution contains a summary of the Krakow meeting on Soft Physics and Fluctuations. It emphasizes both the experimental and the theoretical investigations on correlations/fluctuations and intermittency in multi-particle processes and discusses of the present status of this concept. A clarification of the main open questions in this field of research is now within reach, thanks to the studies presented at the meeting. Summary Talk of the Krakow Workshop on Multiparticle Physics, May, 4-7, 1993. E-mail contact:

  10. Computer-controlled system for the study of oxidase reactions: application to the peroxidase-oxidase oscillator. (United States)

    McDonald, Andrew G; Tipton, Keith F


    An apparatus for the study of bisubstrate oxidase reactions at maintained steady-state substrate concentrations is described, and its specific application to the peroxidase-oxidase biochemical oscillator is reported. Instrument control and data acquisition are provided by custom software written in LabVIEW. The software allows measurement, recording, and control of dissolved oxygen through a Clark-type oxygen electrode, reaction monitoring by a UV/vis spectrophotometer, and controlled substrate delivery by a syringe infusion pump. For peroxidase from horseradish, the optimal pH for oscillatory behavior was found to be in the range 4.5-5.5.

  11. Student questions in urban middle school science communities of practice (United States)

    Groome, Meghan

    This dissertation examines student questions within three Communities of Practice (CoP), all urban middle school science environments. The study analyzed student questions from a sociocultural perspective and used ethnographic research techniques to detail how the CoP's shaped questions in the classroom. In the first study, two case study girls attempted to navigate questioning events that required them to negotiation participation. Their access to participation was blocked by participation frameworks that elevated some students as "gatekeepers" while suppressing the participation of others. The next two studies detail the introduction of written questioning opportunities, one into a public middle school classroom and the other into an informal classroom. In both studies, students responded to the interventions differently, most notable the adoption of the opportunity by female students who do not participate orally. Dissertation-wide findings indicate all students were able to ask questions, but varied in level of cognitive complexity, and the diagnostic interventions were able to identify students who were not known to be "target students", students who asked a high number of questions and were considered "interested in science". Some students' roles were as "gatekeepers" to participation of their peers. Two out of three teachers in the studies reported major shifts in their teaching practice due to the focus on questions and the methods used here have been found to be effective in producing educational research as well as supporting high-need classrooms in prior research. In conclusion, these studies indicate that social factors, including participation frameworks, gender dynamics, and the availability of alternative participation methods, play an important role in how students ask science-related questions. It is recommended that researchers continue to examine social factors that reduce student questions and modify their teaching strategies to facilitate

  12. God in Question: Questioning as a Prerequisite for Theology

    Directory of Open Access Journals (Sweden)

    Martin Kočí


    Full Text Available There are questions that are so important that it is a pity to spoil them with answers. No doubt, the question of God is one of them. Contrary to many presuppositions, theology is not capable of providing us with the final answers in this respect. On the contrary, theology professed as fides quaerens intellectum is an ongoing struggle with questions. Modernity interrupted this paradigm of theological questioning. Theology was withdrawn from the realm of understanding and shifted to the realm of explanation. Modernity brought the univocalization of God. Nonetheless, the attempts to tackle the question of God lead to hegemonic narratives about God. Such narratives are rightly criticized in a postmodern context for their totalizing pretensions. The problem of postmodern criticism is its one-sided emphasis on the apophatic dimension of theological discourse. I propose that theology can go a step further beyond postmodernity. In order to do so, I deal with the Czech philosopher Jan Patočka, who provides an opportunity to rethink God from the perspective of questioning in a new way. Patočka’s insistence on problematicity is the main reading key of his work. In this line of though, I interpret Patočka’s student Tomáš Halík and his thesis about the necessity to take the metaphor of an unknown God into account. I argue that theology must avoid the temptation to remove God from the question and make a well-known God of him. The time has come for theologians to turn their answers back into questions and dwell with them. Bůh jako otázka: Tázání jako předpoklad teologie Některé otázky jsou tak důležité, že je škoda kazit je odpověďmi. Není pochyb, že otázka po Bohu je jednou z nich. Navzdory mnoha předpokladům, ani teologie není schopna poskytnout v tomto ohledu konečné odpovědi. Teologie pojatá jako fides quaerens intellectum je nepřetržitý nekončící zápas s otázkami. Moderna narušila toto paradigma teologick

  13. Questions concerning the nuclear wastes; Les dechets nucleaires en questions

    Energy Technology Data Exchange (ETDEWEB)

    Daures, Pierre [ed.] [Electricite de France (EDF), 75 - Paris (France)


    At present, 75% of the electricity in France is of nuclear origin. Most of French people approve this mode of energy production and agree upon the continuation of the electronuclear sector exploitation. However, as any industry, the nuclear industry produces wastes which constitute a keen preoccupation of the public opinion. The nuclear program, even at its very inception, has provided the appropriate mastering of radioactive wastes by reducing their volume, by conditioning, reprocessing and storing, expressing continually its carefulness for population protection as well as for environment defence against the radiological effects. Pursuing its policy of transparency the EDF demonstrated openness and understanding towards questions raised by anyone. This brochure gives answers to the following 17 questions: -what the nuclear wastes are, which is their origin? - what is their amount? - are the nuclear waste dangerous? - how to treat the nuclear wastes? - are the radioactive waste storage sure? - is the nuclear waste transportation sure? - are these solutions sure? - why searches for long-lived radioactive wastes? - what is transmutation? - shall we bequeath to the next generations our nuclear wastes? - are there particular problems in nuclear power plant decommissioning? - what the wastes issued from decommissioning become? - are the costs of reprocessing and decommissioning taken into account in the price of the kWh? - were the nuclear wastes taken into account since the nuclear program inception? - who manages the nuclear wastes? - why France accepted the reprocessing of nuclear wastes produced in foreign countries? - is there an international policy for nuclear wastes?.

  14. The Membrane Modulates Internal Proton Transfer in Cytochrome c Oxidase

    DEFF Research Database (Denmark)

    Öjemyr, Linda Nasvik; Ballmoos, Christoph von; Faxén, Kristina


    The functionality of membrane proteins is often modulated by the surrounding membrane. Here, we investigated the effect of membrane reconstitution of purified cytochrome c oxidase (CytcO) on the kinetics and thermodynamics of internal electron and proton-transfer reactions during O-2 reduction...

  15. Inhibitory activity of xanthine oxidase by fractions Crateva adansonii

    Directory of Open Access Journals (Sweden)

    A Abdullahi


    Conclusions: Enzyme inhibition mechanism indicated that the mode of inhibition was of a mixed type. Our findings suggest that the therapeutic use of these plants may be due to the observed Xanthine oxidase inhibition, thereby supporting their use in traditional folk medicine against inflammatory-related diseases, in particular, gout.

  16. Low activation barriers characterize intramolecular electron transfer in ascorbate oxidase

    DEFF Research Database (Denmark)

    Farver, O; Pecht, I


    Anaerobic reduction kinetics of the zucchini squash ascorbate oxidase (AO; L-ascorbate:oxygen oxidoreductase, EC by pulse radiolytically produced CO2- radical ions were investigated. Changes in the absorption bands of type 1 [Cu(II)] (610 nm) and type 3 [Cu(II)] (330 nm) were monitored...

  17. Semicarbazide-sensitive amine oxidase (SSAO): from cell to circulation

    NARCIS (Netherlands)

    F. Boomsma (Frans); H. Hut; U. Bagghoe; A.H. van den Meiracker (Anton)


    textabstractSemicarbazide-sensitive amine oxidase (SSAO) is a multi-functional enzyme widely present in nature. It converts primary amines into their corresponding aldehydes, while generating H(2)O(2) and NH(3). In mammals, SSAO circulates in plasma, while a membrane-bound form (of

  18. Purification of gibberellin sub 53 -oxidase from spinach

    Energy Technology Data Exchange (ETDEWEB)

    Wilson, T.M.; Zeevaart, J.A.D. (Michigan State Univ., East Lansing (USA))


    Spinach is a long-day rosette plants, in which stem growth is mediated by gibberellins. It has been shown that two enzymatic steps, GA{sub 53}-oxidase and GA{sub 19}-oxidase, are controlled by light. To develop an understanding into this light regulation, purification of GA{sub 53}-oxidase has been undertaken. The original assay relied on the HPLC separation of the product and substrate, but was considered too slow for the development of a purification scheme. A TLC system was developed which in conjunction with improvements to the assay conditions was sensitive and gave rapid results. The partial purification of the GA{sub 53}-oxidase is achieved by a high speed centrifugation, 40-55% ammonium sulfate precipitation, an hydroxyapatite column, Sephadex G-100 column and an anion exchange FPLC column, Mono Q HR10/10, yielding 1000-fold purification and 15% recovery. Monoclonal antibodies to the protein will be raised and used to further characterize the enzyme.

  19. Semicarbazide-sensitive amine oxidase (SSAO): from cell to circulation

    NARCIS (Netherlands)

    F. Boomsma (Frans); H. Hut; U. Bagghoe; A.H. van den Meiracker (Anton)


    textabstractSemicarbazide-sensitive amine oxidase (SSAO) is a multi-functional enzyme widely present in nature. It converts primary amines into their corresponding aldehydes, while generating H(2)O(2) and NH(3). In mammals, SSAO circulates in plasma, while a membrane-bound form

  20. Molecular dynamics in cytochrome c oxidase Moessbauer spectra deconvolution

    Energy Technology Data Exchange (ETDEWEB)

    Bossis, Fabrizio [Department of Medical Biochemistry, Medical Biology and Medical Physics (DIBIFIM), University of Bari ' Aldo Moro' , Bari (Italy); Palese, Luigi L., E-mail: [Department of Medical Biochemistry, Medical Biology and Medical Physics (DIBIFIM), University of Bari ' Aldo Moro' , Bari (Italy)


    Research highlights: {yields} Cytochrome c oxidase molecular dynamics serve to predict Moessbauer lineshape widths. {yields} Half height widths are used in modeling of Lorentzian doublets. {yields} Such spectral deconvolutions are useful in detecting the enzyme intermediates. -- Abstract: In this work low temperature molecular dynamics simulations of cytochrome c oxidase are used to predict an experimentally observable, namely Moessbauer spectra width. Predicted lineshapes are used to model Lorentzian doublets, with which published cytochrome c oxidase Moessbauer spectra were simulated. Molecular dynamics imposed constraints to spectral lineshapes permit to obtain useful information, like the presence of multiple chemical species in the binuclear center of cytochrome c oxidase. Moreover, a benchmark of quality for molecular dynamic simulations can be obtained. Despite the overwhelming importance of dynamics in electron-proton transfer systems, limited work has been devoted to unravel how much realistic are molecular dynamics simulations results. In this work, molecular dynamics based predictions are found to be in good agreement with published experimental spectra, showing that we can confidently rely on actual simulations. Molecular dynamics based deconvolution of Moessbauer spectra will lead to a renewed interest for application of this approach in bioenergetics.

  1. Inhibition of chickpea seedling copper amine oxidases by tetraethylenepentamine

    Directory of Open Access Journals (Sweden)

    Sona Talaei


    Full Text Available Copper amine oxidases are important enzymes, which contribute to the regulation of mono- and polyamine levels. Each monomer contains one Cu(II ion and 2,4,5-trihydroxyphenylalanine (TPQ as cofactors. They catalyze the oxidative deamination of primary amines to aldehydes with a ping-pong mechanism consisting of a transamination. The mechanism is followed by the transfer of two electrons to molecular oxygen which is reduced to hydrogen peroxide. Inhibitors are important tools in the study of catalytic properties of copper amine oxidases and they also have a wide application in physiological research. In this study, purification of the chickpea seedling amine oxidase, was done via salting out by ammonium sulfate and dialysis, followed by DEAE-cellulose column chromatography. By using the Lineweaver - Burk plot, the Km and Vm of the enzyme were found to be 3.3 mM and 0.95 mmol/min/mg, respectively. In this study, the interaction of chickpea diamino oxidase with tetraethylene- pentamine was studied. Analysis of kinetic data indicated that tetraethylenepentamine (with Ki=0.1 mM inhibits the enzyme by linear mixed inhibitory effect.

  2. Cytokinin oxidase or dehydrogenase? Mechanism of cytokinin degradation in cereals

    DEFF Research Database (Denmark)

    Galuszka, P.; Frebort, I.; Sebela, M.


    wheat enzyme is a monomer 60 kDa, its N-terminal amino-acid sequence shows similarity to hypothetical cytokinin oxidase genes from Arabidopsis thaliana, but not to the enzyme from maize. N-6-isopentenyl-2-(2-hydroxyethylamino)-9-methyladenine is the best substrate from all the cytokinins tested...

  3. The glucose oxidase-peroxidase assay for glucose (United States)

    The glucose oxidase-peroxidase assay for glucose has served as a very specific, sensitive, and repeatable assay for detection of glucose in biological samples. It has been used successfully for analysis of glucose in samples from blood and urine, to analysis of glucose released from starch or glycog...

  4. Copper complexes as biomimetic models of catechol oxidase : mechanistic studies

    NARCIS (Netherlands)

    Koval, Iryna A.


    The research described in this thesis deals with the synthesis of copper(II) complexes with phenol-based or macrocyclic ligands, which can be regarded as model compounds of the active site of catechol oxidase, and with the mechanism of the catalytic oxidation of catechol mediated by these compounds.

  5. Subcellular localization of vanillyl-alcohol oxidase in Penicillium simplicissimum

    NARCIS (Netherlands)

    Fraaije, MW; Sjollema, KA; Veenhuis, M; van Berkel, WJH; Berkel, Willem J.H. van


    Growth of Penicillium simplicissimum on anisyl alcohol, veratryl alcohol or 3-(methoxymethyl)phenol, is associated with the synthesis of relatively large amounts of the hydrogen peroxide producing flavoprotein vanillyl-alcohol oxidase (VAO), Immunocytochemistry revealed that the enzyme has a dual lo

  6. ADP competes with FAD binding in putrescine oxidase

    NARCIS (Netherlands)

    van Hellemond, Erik W.; Mazon, Hortense; Heck, Albert J.; van den Heuvel, Robert H. H.; Heuts, Dominic P. H. M.; Janssen, Dick B.; Fraaije, Marco W.


    Putrescine oxidase from Rhodococcus erythropolis NCIMB 11540 (PuORh) is a soluble homodimeric flavoprotein of 100 kDa, which catalyzes the oxidative deamination of putrescine and some other aliphatic amines. The initial characterization of PuORh uncovered an intriguing feature: the enzyme appeared t

  7. Looking for an Alternative. (United States)

    Kennedy, Jack


    Argues that high school newspapers might do well to create stronger ties with alternative weeklies. Discusses issues of niche marketing, alternative content, and alternative presentation. Notes that high school papers could learn a lot from alternative newspapers. (SR)

  8. Ten questions about systems biology

    DEFF Research Database (Denmark)

    Joyner, Michael J; Pedersen, Bente K


    In this paper we raise 'ten questions' broadly related to 'omics', the term systems biology, and why the new biology has failed to deliver major therapeutic advances for many common diseases, especially diabetes and cardiovascular disease. We argue that a fundamentally narrow and reductionist...... to understand how whole animals adapt to the real world. We argue that a lack of fluency in these concepts is a major stumbling block for what has been narrowly defined as 'systems biology' by some of its leading advocates. We also point out that it is a failure of regulation at multiple levels that causes many...


    Institute of Scientific and Technical Information of China (English)



    This paper addresses problems common in the listening classes, namely, lack of motivation, orientation and sense of achievement and the presence of pressure, fear and frustration. The author examines different factors in the teaching of listening and poses 6 questions that might help teachers to reflect on their teaching. The conclusion is that the aim of listening class is to develop rather than test listening strategies and skills; that students need to be provided with communicative tasks before and while listening; that materials should be realistic, varied and graded ; that listening should be integrated with other skills development; and finally, that learner autonomy provides the ultimate solution to listening problems.

  10. Searching for cognitive enhancement in the Morris water maze: better and worse performance in D-amino acid oxidase knockout (Dao(-/-)) mice. (United States)

    Pritchett, David; Taylor, Amy M; Barkus, Christopher; Engle, Sandra J; Brandon, Nicholas J; Sharp, Trevor; Foster, Russell G; Harrison, Paul J; Peirson, Stuart N; Bannerman, David M


    A common strategy when searching for cognitive-enhancing drugs has been to target the N-methyl-d-aspartate receptor (NMDAR), given its putative role in synaptic plasticity and learning. Evidence in favour of this approach has come primarily from studies with rodents using behavioural assays like the Morris water maze. D-amino acid oxidase (DAO) degrades neutral D-amino acids such as D-serine, the primary endogenous co-agonist acting at the glycine site of the synaptic NMDAR. Inhibiting DAO could therefore provide an effective and viable means of enhancing cognition, particularly in disorders like schizophrenia, in which NMDAR hypofunction is implicated. Indirect support for this notion comes from the enhanced hippocampal long-term potentiation and facilitated water maze acquisition of ddY/Dao(-) mice, which lack DAO activity due to a point mutation in the gene. Here, in Dao knockout (Dao(-/-) ) mice, we report both better and worse water maze performance, depending on the radial distance of the hidden platform from the side wall of the pool. Dao(-/-) mice displayed an increased innate preference for swimming in the periphery of the maze (possibly due to heightened anxiety), which facilitated the discovery of a peripherally located platform, but delayed the discovery of a centrally located platform. By contrast, Dao(-/-) mice exhibited normal performance in two alternative assays of long-term spatial memory: the appetitive and aversive Y-maze reference memory tasks. Taken together, these results question the proposed relationship between DAO inactivation and enhanced long-term associative spatial memory. They also have generic implications for how Morris water maze studies are performed and interpreted.

  11. No question about exciting questions in cell biology.

    Directory of Open Access Journals (Sweden)

    Thomas D Pollard


    Full Text Available Although we have a good grasp of many important processes in cell biology, including knowledge of many molecules involved and how they interact with each other, we still do not understand most of the dynamical features that are the essence of living systems. Fortunately, we now have the ability to dissect biological systems in enough detail to understand their dynamics, including the use of mathematical models to account for past observations and predict future experiments. This deep level of mechanistic understanding should be our goal—not simply to satisfy our scientific curiosity, but also to understand the causes of disease well enough to predict risks, make early diagnoses, and treat effectively. Many big questions remain to be answered before we reach this goal of understanding cellular dynamics.

  12. No question about exciting questions in cell biology. (United States)

    Pollard, Thomas D


    Although we have a good grasp of many important processes in cell biology, including knowledge of many molecules involved and how they interact with each other, we still do not understand most of the dynamical features that are the essence of living systems. Fortunately, we now have the ability to dissect biological systems in enough detail to understand their dynamics, including the use of mathematical models to account for past observations and predict future experiments. This deep level of mechanistic understanding should be our goal—not simply to satisfy our scientific curiosity, but also to understand the causes of disease well enough to predict risks, make early diagnoses, and treat effectively. Many big questions remain to be answered before we reach this goal of understanding cellular dynamics.

  13. Enzyme-Mediated Conversion of Flavin Adenine Dinucleotide (FAD) to 8-Formyl FAD in Formate Oxidase Results in a Modified Cofactor with Enhanced Catalytic Properties. (United States)

    Robbins, John M; Souffrant, Michael G; Hamelberg, Donald; Gadda, Giovanni; Bommarius, Andreas S


    Flavins, including flavin adenine dinucleotide (FAD), are fundamental catalytic cofactors that are responsible for the redox functionality of a diverse set of proteins. Alternatively, modified flavin analogues are rarely found in nature as their incorporation typically results in inactivation of flavoproteins, thus leading to the disruption of important cellular pathways. Here, we report that the fungal flavoenzyme formate oxidase (FOX) catalyzes the slow conversion of noncovalently bound FAD to 8-formyl FAD and that this conversion results in a nearly 10-fold increase in formate oxidase activity. Although the presence of an enzyme-bound 8-formyl FMN has been reported previously as a result of site-directed mutagenesis studies of lactate oxidase, FOX is the first reported case of 8-formyl FAD in a wild-type enzyme. Therefore, the formation of the 8-formyl FAD cofactor in formate oxidase was investigated using steady-state kinetics, site-directed mutagenesis, ultraviolet-visible, circular dichroism, and fluorescence spectroscopy, liquid chromatography with mass spectrometry, and computational analysis. Surprisingly, the results from these studies indicate not only that 8-formyl FAD forms spontaneously and results in the active form of FOX but also that its autocatalytic formation is dependent on a nearby arginine residue, R87. Thus, this work describes a new enzyme cofactor and provides insight into the little-understood mechanism of enzyme-mediated 8α-flavin modifications.

  14. Les questions de migrations internationales (United States)

    Samman, Mouna Liliane


    International migrations have growing implications for both countries of origin and countries of destination. In the latter, the presence of foreigners and of members of their families today creates problems of integration, causes argument and brings mounting xenophobia. Paralleling political, economic and social measures taken by public authorities to respond to these difficulties, education needs to assist in defusing the resulting social tensions by preparing the minds of learners and helping to develop new attitudes. In particular, when educational programmes address questions of international migration, these should be treated in the framework of historical evolution so that their real significance and their true temporal and spatial dimensions become apparent. It is also important that the growing interdependence between countries should be made plain, that national history should be placed in its international context, and that the true consequences of these developments should be made clear. In this context, learners need to be acquainted with Human Rights, thereby stressing universal moral values and the role of the individual. Lastly, questions relating to international migration are usually presented in the media in a selective and partial manner, and the young people who take in this information often accept the hasty judgments which are made of situations as proven facts. This is why all teaching about international migration needs to be considered or reconsidered in the light of the complementary or competing actions of the media.

  15. Pisa Question and Reasoning Skill

    Directory of Open Access Journals (Sweden)

    Ersoy Esen


    Full Text Available The objective of the study is to determine the level of the reasoning skills of the secondary school students. This research has been conducted during the academic year of 2015-2016 with the participation of 51 students in total, from a province in the Black Sea region of Turkey by using random sampling method. Case study method has been used in this study, since it explains an existing situation. In this study, content analysis from the qualitative research methods was carried out. In order to ensure the validity of the scope, agreement percentage formula was used and expert opinions were sought.The problem named Holiday from the Chapter 1 of the normal units in Problem Solving Questions from PISA (Program for International Student Assessments [35] are used as the data collection tool for the study. The problem named Holiday consists of two questions. Applied problems were evaluated according to the mathematical reasoning stages of TIMSS (2003. The findings suggest that the students use proportional reasoning while solving the problems and use the geometric shapes to facilitate the solution of the problem. When they come across problems related to each other, it is observed that they create connections between the problems based on the results of the previous problem. In conclusion, the students perform crosscheck to ensure that their solutions to the problems are accurate.

  16. Are Quantum Theory Questions Epistemic?

    Directory of Open Access Journals (Sweden)

    Viviana Yaccuzzi Polisena


    Full Text Available How to displace-move quantum theory [Ǭ] questions-problems to philosophy? Seeing the collapse of our society’s cultural-intellectual-morals, the philosophy of the 21st century has to contribute to the formation of new principles-formalisms: the big task of the contemporary philosophy ©] is to innovate, to transform the building of the knowledge! Which is the role of the contemporary philosopher? (Noam Chomsky. Building science so that it is more human, out of the scientific mercantilism so that it does not continue transgressing that which is most precious: the thought-life. The ideas that I propose demand a deep cultural-epistemiologicscientific-philosophical-ethical rethinking that goes from quantum entities up to life in society. The starting idea is «the quantum [Ǭ], the paradigm of the contemporary science ©]» (Bernard D’Espagnat. I propose to displace-move questions of the quantum theory [Ǭ]: spin, measure, layering to the field of philosophy (φ to build generic symbols. Can the contemporary episteme model the collapse of the ? For a philosopher, can understanding the importance and the behaviour of the spin bring something new to philosophy ? Can information of the states of the spin be used to observe in a holographic way the pattern energy-information contained in the quantum entities? Is quantum [Ǭ] physics mechanical?

  17. Evaluative Conditioning: The "How" Question. (United States)

    Jones, Christopher R; Olson, Michael A; Fazio, Russell H


    Evaluative conditioning (EC) refers to attitude formation or change toward an object due to that object's mere co-occurrence with another valenced object or objects. This chapter focuses on the "how" question, that is, the question of what cognitive processes intervene between mere co-occurrence and attitude formation or change. Though EC has typically been thought of as occurring through a single, albeit contentious, mechanism, we begin by pointing out that both the heterogeneity of EC methodologies and the abundance of inconsistent results suggest that multiple processes with different characteristics can produce EC. We describe how the earliest posited process of EC, Pavlovian conditioning or signal learning, is a valid mechanism of EC that appears to have operated in some experiments but is unlikely to have operated in others and also cannot account for various EC findings. We describe other mechanisms of EC, when they can be expected to occur, and what characteristics they have. We particularly focus our attention on a process model of EC we have recently introduced, the implicit misattribution model. Finally, we describe the implications of a multi-process view of EC, which we argue can help resolve theoretical controversies and further the application of EC as a practical intervention for influencing attitudes in various domains.

  18. Cyanobacterial lactate oxidases serve as essential partners of N2-fixation and evolved to photorespiratory glycolate oxidases in plants

    NARCIS (Netherlands)

    Hackenberg, C.; Kern, R.; Hüge, J.; Stal, L.J.; Tsuji, Y.; Kopka, J.; Shiraiwa, Y.; Bauwe, H.; Hagemann, M.


    Glycolate oxidase (GOX) is an essential enzyme involved in photorespiratory metabolism in plants. In cyanobacteria and green algae, the corresponding reaction is catalyzed by glycolate dehydrogenases (GlcD). The genomes of N2-fixing cyanobacteria, such as Nostoc PCC 7120 and green algae, appear to h

  19. Cyanobacterial lactate oxidases serve as essential partners in N2-fixation and evolved into photorespiratory glycolate oxidases in plants.

    NARCIS (Netherlands)

    Hackenberg, C.; Kern, R.; Hüge, J; Stal, L.J.; Tsuji, Y.; Kopka, J.; Shiraiwa, Y.; Bauwe, H.; Hagemann, M.


    Glycolate oxidase (GOX) is an essential enzyme involved in photorespiratory metabolism in plants. In cyanobacteria and green algae, the corresponding reaction is catalyzed by glycolate dehydrogenases (GlcD). The genomes of N2-fixing cyanobacteria, such as Nostoc PCC 7120 and green algae, appear to

  20. Cyanobacterial lactate oxidases serve as essential partners of N2-fixation and evolved to photorespiratory glycolate oxidases in plants

    NARCIS (Netherlands)

    Hackenberg, C.; Kern, R.; Hüge, J.; Stal, L.J.; Tsuji, Y.; Kopka, J.; Shiraiwa, Y.; Bauwe, H.; Hagemann, M.


    Glycolate oxidase (GOX) is an essential enzyme involved in photorespiratory metabolism in plants. In cyanobacteria and green algae, the corresponding reaction is catalyzed by glycolate dehydrogenases (GlcD). The genomes of N2-fixing cyanobacteria, such as Nostoc PCC 7120 and green algae, appear to

  1. Cyanobacterial lactate oxidases serve as essential partners in N2-fixation and evolved into photorespiratory glycolate oxidases in plants.

    NARCIS (Netherlands)

    Hackenberg, C.; Kern, R.; Hüge, J; Stal, L.J.; Tsuji, Y.; Kopka, J.; Shiraiwa, Y.; Bauwe, H.; Hagemann, M.


    Glycolate oxidase (GOX) is an essential enzyme involved in photorespiratory metabolism in plants. In cyanobacteria and green algae, the corresponding reaction is catalyzed by glycolate dehydrogenases (GlcD). The genomes of N2-fixing cyanobacteria, such as Nostoc PCC 7120 and green algae, appear to h

  2. Cytochemical Studies on the Localization of Methanol Oxidase and Other Oxidases in Peroxisomes of Methanol-Grown Hansenula polyrnorpha

    NARCIS (Netherlands)

    Veenhuis, M.; Dijken, J.P. van; Harder, W.


    The localization of methanol oxidase activity in cells of methanol-limited chemostat cultures of the yeast Hansenula polymorpha has been studied with different cytochemical staining techniques. The methods were based on enzymatic or chemical trapping of the hydrogen peroxide produced by the enzyme d

  3. Phenol oxidase activity in secondary transformed peat-moorsh soils (United States)

    Styła, K.; Szajdak, L.


    The chemical composition of peat depends on the geobotanical conditions of its formation and on the depth of sampling. The evolution of hydrogenic peat soils is closely related to the genesis of peat and to the changes in water conditions. Due to a number of factors including oscillation of ground water level, different redox potential, changes of aerobic conditions, different plant communities, and root exudes, and products of the degradation of plant remains, peat-moorsh soils may undergo a process of secondary transformation conditions (Sokolowska et al. 2005; Szajdak et al. 2007). Phenol oxidase is one of the few enzymes able to degrade recalcitrant phenolic materials as lignin (Freeman et al. 2004). Phenol oxidase enzymes catalyze polyphenol oxidation in the presence of oxygen (O2) by removing phenolic hydrogen or hydrogenes to from radicals or quinines. These products undergo nucleophilic addition reactions in the presence or absence of free - NH2 group with the eventual production of humic acid-like polymers. The presence of phenol oxidase in soil environments is important in the formation of humic substances a desirable process because the carbon is stored in a stable form (Matocha et al. 2004). The investigations were carried out on the transect of peatland 4.5 km long, located in the Agroecological Landscape Park host D. Chlapowski in Turew (40 km South-West of Poznań, West Polish Lowland). The sites of investigation were located along Wyskoć ditch. The following material was taken from four chosen sites marked as Zbechy, Bridge, Shelterbelt and Hirudo in two layers: cartel (0-50cm) and cattle (50-100cm). The object of this study was to characterize the biochemical properties by the determination of the phenol oxidize activity in two layers of the four different peat-moors soils used as meadow. The phenol oxidase activity was determined spectrophotometrically by measuring quinone formation at λmax=525 nm with catechol as substrate by method of Perucci

  4. Appearance questions can be misleading

    DEFF Research Database (Denmark)

    Hansen, Mikkel; Markman, Ellen M.


    Preschoolers' success on the appearance-reality task is a milestone in theory-of-mind development. On the standard task children see a deceptive object, such as a sponge that looks like a rock, and are asked, "What is this really?" and "What does this look like?" Children below 4 1/2 years of age...... fail saying that the object not only is a sponge but also looks like a sponge. We propose that young children's difficulty stems from ambiguity in the meaning of "looks like." This locution can refer to outward appearance ("Peter looks like Paul") but in fact often refers to likely reality ("That looks...... like Jim"). We propose that "looks like" is taken to refer to likely reality unless the reality is already part of the common ground of the conversation. Because this joint knowledge is unclear to young children on the appearance-reality task, they mistakenly think the appearance question is about...

  5. Questions and Answers about BSP

    Directory of Open Access Journals (Sweden)

    D.B. Skillicorn


    Full Text Available Bulk Synchronous Parallelism (BSP is a parallel programming model that abstracts from low-level program structures in favour of supersteps. A superstep consists of a set of independent local computations, followed by a global communication phase and a barrier synchronisation. Structuring programs in this way enables their costs to be accurately determined from a few simple architectural parameters, namely the permeability of the communication network to uniformly-random traffic and the time to synchronise. Although permutation routing and barrier synch ronisations are widely regarded as inherently expensive, this is not the case. As a result, the structure imposed by BSP does not reduce performance, while bringing considerable benefits for application building. This paper answers the most common questions we are asked about BSP and justifies its claim to be a major step forward in parallel programming.

  6. Questioning Strategy in English Teaching

    Institute of Scientific and Technical Information of China (English)



    Learner autonomy has been a hot issue discussed by educators in recent years.Because it is believed that the ulti-mate goal of educating is to help students learn autonomously and actively not only in school but also,after graduation,for their whole life.Many people misunderstand the meaning of learner autonomy as learners learn something totally independent from teacher or learn after class.This is not the truth;teacher is indispensible to promote learner autonomy.Therefore,English teacher bears a great respon-sibility of fostering and promoting learner autonomy in the classroom.Questioning is the most frequently used and easiest method in the various teaching methods.

  7. Questions That Engage Students in Mathematical Thinking. (United States)

    Ilaria, Daniel R.

    Teacher questions are an important part of a student-inquiry classroom. This research examines two different student-centered settings to determine the teacher questions that engaged students in mathematical thinking. It reports on questions asked in both a research setting and a high school classroom. Discursive and retracing questions are…

  8. On Effective Questioning in English Classes

    Institute of Scientific and Technical Information of China (English)



    Teachers' questioning has traditionally been viewed as an important constituent of teacher talk and the core of effective teaching in classroom context.Questioning is the basic means used by teachers to challenge students to think,appraise teaching results and help students to realize their desired goals,thus,playing a key role in the second language acquisition.This paper will first introduce the definition and types of teachers'questioning and the criteria for effective questioning.It will also give some practical suggestions for practice effective questioning,focusing on good questions,wait-time,feedback and allocation of questions.

  9. Socrates' questions: a focus for nursing. (United States)

    Bunkers, Sandra S


    This column focuses on the philosophical dialogue originated by Socrates. Six questions that Socrates would ask the ancient Greeks are explored in discussing a book written by Phillips entitled Six Questions of Socrates. These questions were: What is virtue? What is moderation? What is justice? What is good? What is courage? What is piety? A human becoming perspective is used as a lens to view the discussion on these questions and the question is posed, "What would it be like to frame discussions on health and quality of life around Socrates' questions?" Parse's teaching-learning processes are presented as a means of creating an environment where dialogue on these questions can occur.

  10. The Recognition of Glycolate Oxidase Apoprotein with Flavin Analogs in Higher Plants

    Institute of Scientific and Technical Information of China (English)

    Wei-Jun WANG; Jing-Quan HUANG; Chong YANG; Jiu-Jiu HUANG; Ming-Qi LI


    The dependence of glycolate oxidase apoprotein (apoGO) activity on flavin analogs was surveyed in 9 higher plants from 7 families. Activities of all apoGOs depended not only on flavin mononucleotide (FMN) but also on flavin adenine dinucleotide (FAD), but not on riboflavin. The kinetic analysis showed that FMN was the optimum cofactor for apoGO from leaves ofBrassica campestris. In plant kingdom,FMN, FAD and riboflavin are three flavin analogs with very similar structure, and they could coexist and be inter-converted from each other, so the question is how the apoprotein of glycolate oxidase (GO) recognized these flavin analogs. No inhibition effect of riboflavin on the activity ofapoGO with FMN or FAD was found and no obvious quenching of riboflavin or apoGO protein fluorescence was detected with the addition of apoGO or riboflavin, respectively. These results indicated that riboflavin did not bind to apoGO tightly like FMN and FAD. Inorganic phosphate (Pi) did inhibit the activity of GO, and kinetic analysis revealed that this inhibition was caused by the competitive binding to apoGO between Pi and FMN. This competitive binding was further confirmed by the inhibition of Pi to the quenching of FMN and apoGO protein fluorescence with apoGO and FMN, respectively. It was suggested that the 5′-phosphate group of FMN or FAD may play a key role in the recognition and binding of riboflavin analog cofactors with apoGO.

  11. 77 FR 50372 - Guidance for Industry: Questions and Answers Regarding the Final Rule, Prevention of Salmonella... (United States)


    ... HUMAN SERVICES Food and Drug Administration 21 CFR Parts 16 and 118 Guidance for Industry: Questions and... rule, including questions and answers on compliance dates; coverage; definitions; SE prevention... confer any rights for or on any person and does not operate to bind FDA or the public. An alternate...

  12. Properties of purified recombinant human polyamine oxidase, PAOh1/SMO. (United States)

    Wang, Yanlin; Murray-Stewart, Tracy; Devereux, Wendy; Hacker, Amy; Frydman, Benjamin; Woster, Patrick M; Casero, Robert A


    The discovery of an inducible oxidase whose apparent substrate preference is spermine indicates that polyamine catabolism is more complex than that originally proposed. To facilitate the study of this enzyme, the purification and characterization of the recombinant human PAOh1/SMO polyamine oxidase are reported. Purified PAOh1/SMO oxidizes both spermine (K(m)=1.6 microM) and N(1)-acetylspermine (K(m)=51 microM), but does not oxidize spermidine. The purified human enzyme also does not oxidize eight representative antitumor polyamine analogues; however, specific oligamine analogues were found to be potent inhibitors of the oxidation of spermine by PAOh1/SMO. The results of these studies are consistent with the hypothesis that PAOh1/SMO represents a new addition to the polyamine metabolic pathway that may represent a new target for antineoplastic drug development.

  13. Physiological roles of plastid terminal oxidase in plant stress responses

    Indian Academy of Sciences (India)

    Xin Sun; Tao Wen


    The plastid terminal oxidase (PTOX) is a plastoquinol oxidase localized in the plastids of plants. It is able to transfer electrons from plastoquinone (PQ) to molecular oxygen with the formation of water. Recent studies have suggested that PTOX is beneficial for plants under environmental stresses, since it is involved in the synthesis of photoprotective carotenoids and chlororespiration, which could potentially protect the chloroplast electron transport chain (ETC) from over-reduction. The absence of PTOX in plants usually results in photo-bleached variegated leaves and impaired adaptation to environment alteration. Although PTOX level and activity has been found to increase under a wide range of stress conditions, the functions of plant PTOX in stress responses are still disputed now. In this paper, the possible physiological roles of PTOX in plant stress responses are discussed based on the recent progress.

  14. Involvement of NADH Oxidase in Biofilm Formation in Streptococcus sanguinis.

    Directory of Open Access Journals (Sweden)

    Xiuchun Ge

    Full Text Available Biofilms play important roles in microbial communities and are related to infectious diseases. Here, we report direct evidence that a bacterial nox gene encoding NADH oxidase is involved in biofilm formation. A dramatic reduction in biofilm formation was observed in a Streptococcus sanguinis nox mutant under anaerobic conditions without any decrease in growth. The membrane fluidity of the mutant bacterial cells was found to be decreased and the fatty acid composition altered, with increased palmitic acid and decreased stearic acid and vaccenic acid. Extracellular DNA of the mutant was reduced in abundance and bacterial competence was suppressed. Gene expression analysis in the mutant identified two genes with altered expression, gtfP and Idh, which were found to be related to biofilm formation through examination of their deletion mutants. NADH oxidase-related metabolic pathways were analyzed, further clarifying the function of this enzyme in biofilm formation.

  15. Reincarnation Revisited: Question format and the distribution of belief in reincarnation in survey research


    Siegers, Pascal


    Comparing frequency of belief in reincarnation from different international survey projects (RAMP, EVS, ISSP) reveals differences of about 15 to 20 percent depending on the specific question format. If single binary questions are used, then belief in reincarnation is more often reported than if a forced-choice question is used which offers respondents alternatives to belief in reincarnation (e.g. resurrection). One possible explanation for this result is that respondents confuse reincarnation...


    Directory of Open Access Journals (Sweden)

    O. Brenifier


    Full Text Available The aim of the investigation is to disclose the possible philosophicalconversation with the child.Methods. The author uses general scientific research methods, including observation and interviews, philosophical analysis.Results and scientific novelty. The author reveals the essence of philosophical conversations with the child, calls the main reasons for the extinction of the children’s curiosity, illustrating examples of incorrect behavior of adults to communicate with children. It is recommended how to be responsible for children’s issues. The article discusses the main reasons for the extinction of the children’s curiosity by illustrating examples of an erroneous behaviour of adults in dealing with children. It is shown that if the teacher does not find a systematic way to engage children in the essential discussion, the children most likely will not learn how to contemplate seriously. The author gives detailed guidance how to answer children’s questions.Practical significance. The article may be of interest to parents, teachers, experts in the field of psychology of creativity, post-graduates and organizers of independent activity of students of higher education institutions.

  17. Frequently Asked Questions: The Higgs!

    CERN Multimedia

    CERN Bulletin


    Why have we tried so hard to find the Higgs particle? How does the Higgs mechanism work? What is the difference in physics between strong evidence and a discovery? Why do physicists speak in terms of "sigmas"? Find out here!   Why have we tried so hard to find the Higgs particle? Because it could be the answer to the question: how does Nature decide whether or not to assign mass to particles? All the fundamental particles making up matter – the electron, the quarks, etc. – have masses. Moreover, quantum physics requires that forces are also carried by particles. The W and Z particles that carry the weak force responsible for radioactivity must also have masses, whereas the photon, the carrier of the electromagnetic force, has no mass at all. This is the root of the “Higgs problem”: how to give masses to the fundamental particles and break the symmetry between the massive W and Z and the massless photon? Just assigning masses by hand...

  18. Verum focus and polar questions

    Directory of Open Access Journals (Sweden)

    Ion Giurgea


    Full Text Available We argue that some word order phenomena in Romanian and Sardinian are the result of a checking operation in the left periphery involving verum focus (i.e. focus on the polarity component of the sentence. In particular, this operation accounts for some word order patterns found in polar questions. In Romanian, polarity fronting is realized as head-movement of (V+T to a higher peripheral head which bears a Focus-probe. This licenses VS orders for predications in which VS is not allowed as a neutral order (i-level predicates, iteratives, generics. In Sardinian, an entire phrase headed by the lexical predicate (verbal non-finite form or non-verbal predicate is fronted before the auxiliary. We argue that this order is obtained by two movement operations, head-raising of Aux to Foc and movement of the predicate phrase to SpecFoc. We also present the semantics of polarity focus, distinguishing several types of focus (informational, emphatic, contrastive.

  19. Verum focus and polar questions

    Directory of Open Access Journals (Sweden)

    Ion Giurgea


    Full Text Available We argue that some word order phenomena in Romanian and Sardinian are the result of a checkingoperation in the left periphery involving verum focus (i.e. focus on the polarity component of the sentence.In particular, this operation accounts for some word order patterns found in polar questions. In Romanian,polarity fronting is realized as head-movement of (V+T to a higher peripheral head which bears a Focusprobe.This licenses VS orders for predications in which VS is not allowed as a neutral order (i-levelpredicates, iteratives, generics. In Sardinian, an entire phrase headed by the lexical predicate (verbal nonfiniteform or non-verbal predicate is fronted before the auxiliary. We argue that this order is obtained bytwo movement operations, head-raising of Aux to Foc and movement of the predicate phrase to SpecFoc. Wealso present the semantics of polarity focus, distinguishing several types of focus (informational, emphatic,contrastive.

  20. Deglycosylation of glucose oxidase to improve biosensors and biofuel cells

    Energy Technology Data Exchange (ETDEWEB)

    Prevoteau, Antonin; Courjean, Olivier; Mano, Nicolas [Universite de Bordeaux, CNRS, Centre de Recherche Paul Pascal (CRPP), UPR 8641, Avenue Albert Schweitzer, 33600 Pessac (France)


    We demonstrate that a more efficient redox hydrogel structure can be achieved by engineering the size and the surface charge of the bioelectrocatalyst. Deglycosylated glucose oxidase (GOx) modified electrode exhibits higher current density than native GOx, for the same molar composition of the hydrogel. This improvement is very likely due to a more efficient hydrogel structure rather than a better intrinsic electron transfer between the FAD/FADH{sub 2} redox center and the redox mediator. (author)



    Joseph Baby; Rajan Sheeja S; M.V Jeevitha; S.U Ajisha


    L-Amino acid oxidases are flavoenzymes which catalyze the stereospecific oxidative deamination of an L-amino acid substrate to a corresponding a-ketoacid with hydrogen peroxide and ammonia production. These enzymes, which are widely distributed in many different organisms, exhibit a marked affinity for hydrophobic amino acids, including phenylalanine, tryptophan, tyrosine, and leucine. Snake venom LAAO induces platelet aggregation and cytotoxicity in various cancer cell lines. The enzyme has ...

  2. Nitrogen heterocycles as potential monoamine oxidase inhibitors: Synthetic aspects

    Directory of Open Access Journals (Sweden)

    Pravin O. Patil


    Full Text Available The present review highlights the synthetic methods of monoamine oxidase inhibitors (MAO belonging to a group of nitrogen heterocycles such as pyrazoline, indole, xanthine, oxadiazole, benzimidazole, pyrrole, quinoxaline, thiazole and other related compounds (1990–2012. Moreover, it emphasizes salient findings related to chemical structures and the bioactivities of these heterocycles as MAO inhibitors. The aim of this review is to find out different methods for the synthesis of nitrogen containing heterocycles and their bioactivity related aspects as MAO inhibitors.

  3. Steady state equivalence among autocatalytic peroxidase-oxidase reactions (United States)

    Méndez-González, José; Femat, Ricardo


    Peroxidase-oxidase is an enzymatic reaction that can exhibit dynamical scenarios such as bistability, sustained oscillations, and Shilnikov chaos. In this work, we apply the chemical reaction network theory approach to find kinetic constants such that the associated mass action kinetics ordinary differential equations induced by three four dimensional structurally different enzymatic reaction systems can support the same steady states for several chemical species despite differences in their chemical nature.

  4. A novel fluorogenic probe for monoamine oxidase assays

    Institute of Scientific and Technical Information of China (English)

    You You Lu; Yu Guang Wang; Bin Dai; Yi Qi Dai; Zhao Wang; Zheng Wei Fu; Qing Zhu


    Monoamine oxidase is flavoenzymes, widely distributed in mammals. It is well recognized that MAOs serve an important role in metabolism that they have close relationship with health .Along with the discoveries between MAOs and neurotic disease, more and more studies have been jumped in .In this paper, we design a new probe for assaying the activities of MAOs. The results showed that the probe [7-(3-aminopropoxy)coumarin] is simple, effective and sensitive for MAOB.

  5. Inventory control: cytochrome oxidase assembly regulates mitochondrial translation (United States)

    Mick, David U.; Fox, Thomas D.; Rehling, Peter


    Mitochondria maintain a genome and translation-machinery to synthesize a small subset of subunits of the oxidative phosphorylation system. These organellar gene products must assemble with imported subunits that are encoded in the nucleus to build up functional enzymes. New findings on the early steps in cytochrome oxidase assembly reveal how the mitochondrial translation of its core component Cox1 is directly coupled to the assembly of this respiratory complex. PMID:21179059

  6. FVQA: Fact-based Visual Question Answering. (United States)

    Wang, Peng; Wu, Qi; Shen, Chunhua; Dick, Anthony; Hengel, Anton van den


    Visual Question Answering (VQA) has attracted much attention in both computer vision and natural language processing communities, not least because it offers insight into the relationships between two important sources of information. Current datasets, and the models built upon them, have focused on questions which are answerable by direct analysis of the question and image alone. The set of such questions that require no external information to answer is interesting, but very limited. It excludes questions which require common sense, or basic factual knowledge to answer, for example. Here we introduce FVQA (Fact-based VQA), a VQA dataset which requires, and supports, much deeper reasoning. FVQA primarily contains questions that require external information to answer. We thus extend a conventional visual question answering dataset, which contains image-question-answer triplets, through additional image-question-answer-supporting fact tuples. Each supporting-fact is represented as a structural triplet, such as .

  7. Biocompatibility selenium nanoparticles with an intrinsic oxidase-like activity (United States)

    Guo, Leilei; Huang, Kaixun; Liu, Hongmei


    Selenium nanoparticles (SeNPs) are considered to be the new selenium supplement forms with high biological activity and low toxicity; however, the molecular mechanism by which SeNPs exert the biological function is unclear. Here, we reported that biocompatibility SeNPs possessed intrinsic oxidase-like activity. Using Na2SeO3 as a precursor and glutathione as a reductant, biocompatibility SeNPs were synthesized by the wet chemical reduction method in the presence of bovine serum albumin (BSA). The results of structure characterization revealed that synthesized SeNPs were amorphous red elementary selenium with spherical morphology, and ranged in size from 25 to 70 nm size with a narrow distribution (41.4 ± 6.7 nm). The oxidase-like activity of the as-synthesized SeNPs was tested with 3,3',5,5'-tetramethylbenzidine (TMB) as a substrate. The results indicated that SeNPs could catalyze the oxidization of TMB by dissolved oxygen. These SeNPs showed an optimum catalytic activity at pH 4 and 30 °C, and the oxidase-like activity was higher as the concentration of SeNPs increased and the size of SeNPs decreased. The Michaelis constant ( K m) values and maximal reaction velocity ( V max) of the SeNPs for TMB oxidation were 0.0083 mol/L and 3.042 μmol/L min, respectively.

  8. Inhibitory activity of xanthine oxidase by fractions Crateva adansonii

    Institute of Scientific and Technical Information of China (English)

    Abdullahi A; Kolo MZ; Hamzah RU; Jigam AA; Yahya A; Kabiru AY; Muhammad H; Sakpe S; Adefolalu FS; Isah MC


    Objective: To study the inhibitory effect of various extracts from Crateva adansonii (C. adansonii) used traditionally against several inflammatory diseases such as rheumatism, arthritis, and gout, was investigated on purified bovine milk xanthine oxidase (XO) activity. Methods:Xanthine oxidase inhibitory activity was assayed spectrophotometrically and the degree of enzyme inhibition was determined by measuring the increase in absorbance at 295 nm associated with uric acid formation. Enzyme kinetics was carried out using Lineweaver-Burk plots using xanthine as the substrate. Results: Among the fractions tested, the chloroform fraction exhibited highest potency (IC50 20.2±1.6 μg/mL) followed by the petroleum ether (IC50 30.1±2.2 μg/mL), ethyl acetate (IC50 43.9±1.4 μg/mL) and residual (IC50 98.0±3.3 μg/mL) fractions. The IC50 value of allopurinol used, as the standard was 5.7±0.3 μg/mL. Conclusions: Enzyme inhibition mechanism indicated that the mode of inhibition was of a mixed type. Our findings suggest that the therapeutic use of these plants may be due to the observed Xanthine oxidase inhibition, thereby supporting their use in traditional folk medicine against inflammatory-related diseases, in particular, gout.

  9. Surface characterization and direct bioelectrocatalysis of multicopper oxidases

    Energy Technology Data Exchange (ETDEWEB)

    Ivnitski, Dmitri M., E-mail: ivnitski@unm.ed [Chemical and Nuclear Engineering, University of New Mexico, Albuquerque 87131 (United States)] [Air Force Research Laboratory, AFRL/RXQL, Microbiology and Applied Biochemistry, Tyndall Air Force Base, FL 32403 (United States); Khripin, Constantine [Chemical and Nuclear Engineering, University of New Mexico, Albuquerque 87131 (United States); Luckarift, Heather R. [Air Force Research Laboratory, AFRL/RXQL, Microbiology and Applied Biochemistry, Tyndall Air Force Base, FL 32403 (United States)] [Universal Technology Corporation, 1270 N. Fairfield Road, Dayton, OH 45432 (United States); Johnson, Glenn R. [Air Force Research Laboratory, AFRL/RXQL, Microbiology and Applied Biochemistry, Tyndall Air Force Base, FL 32403 (United States); Atanassov, Plamen, E-mail: plamen@unm.ed [Chemical and Nuclear Engineering, University of New Mexico, Albuquerque 87131 (United States)


    Multicopper oxidases (MCO) have been extensively studied as oxygen reduction catalysts for cathodic reactions in biofuel cells. Theoretically, direct electron transfer between an enzyme and electrode offers optimal energy conversion efficiency providing that the enzyme/electrode interface can be engineered to establish efficient electrical communication. In this study, the direct bioelectrocatalysis of three MCO (Laccase from Trametes versicolor, bilirubin oxidase (BOD) from the fungi Myrothecium verrucaria and ascorbate oxidase (AOx) from Cucurbita sp.) was investigated and compared as oxygen reduction catalysts. Protein film voltammetry and electrochemical characterization of the MCO electrodes showed that DET had been successfully established in all cases. Atomic force microscopy imaging and force measurements indicated that enzyme was immobilized as a monolayer on the electrode surface. Evidence for three clearly separated anodic and cathodic redox events related to the Type 1 (T1) and the trinculear copper centers (T2, T3) of various MCO was observed. The redox potential of the T1 center was strongly modulated by physiological factors including pH, anaerobic and aerobic conditions and the presence of inhibitors.

  10. Mechanisms for suppressing NADPH oxidase in the vascular wall

    Directory of Open Access Journals (Sweden)

    Gregory J Dusting


    Full Text Available Oxidative stress underlies many forms of vascular disease as well as tissue injury following ischemia and reperfusion. The major source of oxidative stress in the artery wall is an NADPH oxidase. This enzyme complex as expressed in vascular cells differs from that in phagocytic leucocytes both in biochemical structure and functions. The crucial flavin-containing catalytic subunits, Nox1 and Nox4, are not found in leucocytes, but are highly expressed in vascular cells and upregulated with vascular remodeling, such as that found in hypertension and atherosclerosis. The difference in catalytic subunits offers the opportunity to develop "vascular specific" NADPH oxidase inhibitors that do not compromise the essential physiological signaling and phagocytic functions carried out by reactive oxygen and nitrogen species. Nitric oxide and targeted inhibitors of NADPH oxidase that block the source of oxidative stress in the vasculature are more likely to prevent the deterioration of vascular function that leads to stroke and heart attack, than are conventional antioxidants. The roles of Nox isoforms in other inflammatory conditions are yet to be explored.

  11. Kinetic mechanism of putrescine oxidase from Rhodococcus erythropolis. (United States)

    Kopacz, Malgorzata M; Heuts, Dominic P H M; Fraaije, Marco W


    Putrescine oxidase from Rhodococcus erythropolis (PuO) is a flavin-containing amine oxidase from the monoamine oxidase family that performs oxidative deamination of aliphatic diamines. In this study we report pre-steady-state kinetic analyses of the enzyme with the use of single- and double-mixing stopped-flow spectroscopy and putrescine as a substrate. During the fast and irreversible reductive half-reaction no radical intermediates were observed, suggesting a direct hydride transfer from the substrate to the FAD. The rate constant of flavin reoxidation depends on the ligand binding; when the imine product was bound to the enzyme the rate constant was higher than with free enzyme species. Similar results were obtained with product-mimicking ligands and this indicates that a ternary complex is formed during catalysis. The obtained kinetic data were used together with steady-state rate equations derived for ping-pong, ordered sequential and bifurcated mechanisms to explore which mechanism is operative. The integrated analysis revealed that PuO employs a bifurcated mechanism due to comparable rate constants of product release from the reduced enzyme and reoxidation of the reduced enzyme-product complex.

  12. Alternative medicine - pain relief (United States)

    Alternative medicine refers to treatments that are used instead of conventional (standard) ones. If you use an alternative ... with conventional medicine or therapy, it is considered complementary therapy. There are many forms of alternative medicine. Acupuncture ...

  13. Proton transfer in ba(3) cytochrome c oxidase from Thermus thermophilus. (United States)

    von Ballmoos, Christoph; Adelroth, Pia; Gennis, Robert B; Brzezinski, Peter


    The respiratory heme-copper oxidases catalyze reduction of O(2) to H(2)O, linking this process to transmembrane proton pumping. These oxidases have been classified according to the architecture, location and number of proton pathways. Most structural and functional studies to date have been performed on the A-class oxidases, which includes those that are found in the inner mitochondrial membrane and bacteria such as Rhodobacter sphaeroides and Paracoccus denitrificans (aa(3)-type oxidases in these bacteria). These oxidases pump protons with a stoichiometry of one proton per electron transferred to the catalytic site. The bacterial A-class oxidases use two proton pathways (denoted by letters D and K, respectively), for the transfer of protons to the catalytic site, and protons that are pumped across the membrane. The B-type oxidases such as, for example, the ba(3) oxidase from Thermus thermophilus, pump protons with a lower stoichiometry of 0.5 H(+)/electron and use only one proton pathway for the transfer of all protons. This pathway overlaps in space with the K pathway in the A class oxidases without showing any sequence homology though. Here, we review the functional properties of the A- and the B-class ba(3) oxidases with a focus on mechanisms of proton transfer and pumping. Copyright © 2012 Elsevier B.V. All rights reserved.

  14. Role of amine oxidase expression to maintain putrescine homeostasis in Rhodococcus opacus. (United States)

    Foster, Alexander; Barnes, Nicole; Speight, Robert; Morris, Peter C; Keane, Mark A


    While applications of amine oxidases are increasing, few have been characterised and our understanding of their biological role and strategies for bacteria exploitation are limited. By altering the nitrogen source (NH4Cl, putrescine and cadaverine (diamines) and butylamine (monoamine)) and concentration, we have identified a constitutive flavin dependent oxidase (EC within Rhodococcus opacus. The activity of this oxidase can be increased by over two orders of magnitude in the presence of aliphatic diamines. In addition, the expression of a copper dependent diamine oxidase (EC was observed at diamine concentrations>1mM or when cells were grown with butylamine, which acts to inhibit the flavin oxidase. A Michaelis-Menten kinetic treatment of the flavin oxidase delivered a Michaelis constant (KM)=190μM and maximum rate (kcat)=21.8s(-1) for the oxidative deamination of putrescine with a lower KM (=60μM) and comparable kcat (=18.2s(-1)) for the copper oxidase. MALDI-TOF and genomic analyses have indicated a metabolic clustering of functionally related genes. From a consideration of amine oxidase specificity and sequence homology, we propose a putrescine degradation pathway within Rhodococcus that utilises oxidases in tandem with subsequent dehydrogenase and transaminase enzymes. The implications of PUT homeostasis through the action of the two oxidases are discussed with respect to stressors, evolution and application in microbe-assisted phytoremediation or bio-augmentation.

  15. Alkylamino derivatives of 4-aminomethylpyridine as inhibitors of copper-containing amine oxidases. (United States)

    Bertini, Vincenzo; Buffoni, Franca; Ignesti, Giovanni; Picci, Nevio; Trombino, Sonia; Iemma, Francesca; Alfei, Silvana; Pocci, Marco; Lucchesini, Francesco; De Munno, Angela


    The first substratelike, reversible inhibitors of different copper amine oxidases (CAOs) with IC50 (M) as low as 2.0 x 10(-8) corresponding to derivatives of 4-aminomethylpyridine with alkoxy (1a-d), alkylthio (2a,b), and alkylamino (3a-e, 4a-j) groups in the positions 3 and 5 have been prepared and studied. The inhibitors 1a-d are active on benzylamine oxidase and semicarbazide-sensitive amine oxidase and are very selective with respect to diamine oxidase, lysyl oxidase, and monoamine oxidases. The inhibitors 2a,b are selective for benzylamine oxidase whereas 2a is also a new type of good substrate of diamine oxidase. The inhibitors 3a-e and 4a-j are substratelike, reversible, nonselective inhibitors of various CAOs including pea seedling amine oxidase and Hansenula polymorpha amine oxidase, whose enzymatic sites are known from X-ray structure determinations. The inhibitors 3b,c and 4b,c are excellent substratelike tools for studies correlating CAOs that afford crystals suitable for X-ray structure determinations with CAOs from mammals.

  16. Characterization of Truncated Tumor-Associated NADH Oxidase (ttNOX) (United States)

    Karr, Laurel J.; Malone, Christine C.; Burk, Melissa; Moore, Blake P.; Achari, Aniruddha; Curreri, Peter A. (Technical Monitor)


    Bacterial, plant and animal cells possess novel surface proteins that exhibit both NADH oxidation (NOX) or hydroquinone and protein disulfide-thiol interchange. These enzymatic activities alternate to yield oscillating patterns wjth period lengths of approximately 24 minutes. The catalytic period of NOX proteins are temperature compensated and gravity responsive. We report the cloning, expression and characterization of truncated tumor-associated NADH oxidase (ttNOX), in which the membrane spanning region has been deleted. The cDNA (originated from HeLa cells) was cloned into pET-34b and pET-14b (Novagen) vectors for E. coli expression. Optimized expression and purification protocols yielded greater than 300mg per liter of culture with greater than 95% purity. Circular dichroism data was collected from a 2.7mg/ml solution in a 0.1mm cuvette with variable scanning using an Olis RSM CD spectrophotometer. The ellipticity values were scanned from 190 to 260nm. The spectra recorded have characteristics for alpha proteins with band maxima at 216nm and a possible shoulder at 212nm at 12OC and 250 C. Protein crystal screens are in progress and, to date, only small crystals have been observed. The regular periodic oscillatory change in the ttNOX protein is indicative of a possible time-keeping functional role. A single protein possessing alternating catalytic activities, with a potential biological clock function, is unprecedented and structural determination is paramount to understanding this role.

  17. New Hybrid Algorithm for Question Answering

    Directory of Open Access Journals (Sweden)

    Jaspreet Kaur


    Full Text Available With technical advancement, Question Answering has emerged as the main area for the researchers. User is provided with specific answers instead of large number of documents or passages in question answering. Question answering proposes the solution to acquire efficient and exact answers to user question asked in natural language rather than language query. The major goal of this paper is to develop a hybrid algorithm for question answering. For this task different question answering systems for different languages were studied. After deep study, we are able to develop an algorithm that comprises the best features from excellent systems. An algorithm developed by us performs well.

  18. Replacement of a terminal cytochrome c oxidase by ubiquinol oxidase during the evolution of acetic acid bacteria. (United States)

    Matsutani, Minenosuke; Fukushima, Kota; Kayama, Chiho; Arimitsu, Misato; Hirakawa, Hideki; Toyama, Hirohide; Adachi, Osao; Yakushi, Toshiharu; Matsushita, Kazunobu


    The bacterial aerobic respiratory chain has a terminal oxidase of the heme-copper oxidase superfamily, comprised of cytochrome c oxidase (COX) and ubiquinol oxidase (UOX); UOX evolved from COX. Acetobacter pasteurianus, an α-Proteobacterial acetic acid bacterium (AAB), produces UOX but not COX, although it has a partial COX gene cluster, ctaBD and ctaA, in addition to the UOX operon cyaBACD. We expressed ctaB and ctaA genes of A. pasteurianus in Escherichia coli and demonstrated their function as heme O and heme A synthases. We also found that the absence of ctaD function is likely due to accumulated mutations. These COX genes are closely related to other α-Proteobacterial COX proteins. However, the UOX operons of AAB are closely related to those of the β/γ-Proteobacteria (γ-type UOX), distinct from the α/β-Proteobacterial proteins (α-type UOX), but different from the other γ-type UOX proteins by the absence of the cyoE heme O synthase. Thus, we suggest that A. pasteurianus has a functional γ-type UOX but has lost the COX genes, with the exception of ctaB and ctaA, which supply the heme O and A moieties for UOX. Our results suggest that, in AAB, COX was replaced by β/γ-Proteobacterial UOX via horizontal gene transfer, while the COX genes, except for the heme O/A synthase genes, were lost. Copyright © 2014 Elsevier B.V. All rights reserved.

  19. Frequently Asked Questions about Bunion Surgery (United States)

    ... A | Print | Share Frequently Asked Questions About Bunion Surgery Here are some frequently asked questions (FAQs) and ... best for you. 5. How can I avoid surgery? Sometimes observation of the bunion is all that ...

  20. Approaches to Exploring Category Information for Question Retrieval in Community Question-Answer Archives

    DEFF Research Database (Denmark)

    Cao, Xin; Cong, Gao; Cui, Bin


    of CQA services, question retrieval in a CQA archive aims to retrieve historical question-answer pairs that are relevant to a query question. This article presents several new approaches to exploiting the category information of questions for improving the performance of question retrieval...

  1. Questioning as Thinking: A Metacognitive Framework (United States)

    Wilson, Nance S.; Smetana, Linda


    The purpose of this article is to present Questioning as Thinking (QAT), a metacognitive framework that helps teachers and students focus on performance. QAT combines components of three widely-used strategies: Think-Alouds, Question Answer Relationships (QAR), and Self-Questioning. Together, these three strategies give students and teachers the…

  2. IMS Learning Design Frequently Asked Questions

    NARCIS (Netherlands)

    Tattersall, Colin; Manderveld, Jocelyn; Hummel, Hans; Sloep, Peter; Koper, Rob; De Vries, Fred


    This list of frequently asked questions was composed on the basis of questions asked of the Educational Technology Expertise Centrum. The questions addessed are: Where can I find the IMS Learning Design Specification? What is meant by the phrase “Learning Design”? What is the IMS LD Specification ab

  3. Questions and Answers About Nuclear Power Plants. (United States)

    Environmental Protection Agency, Washington, DC.

    This pamphlet is designed to answer many of the questions that have arisen about nuclear power plants and the environment. It is organized into a question and answer format, with the questions taken from those most often asked by the public. Topics include regulation of nuclear power sources, potential dangers to people's health, whether nuclear…

  4. Questions That Science Teachers Find Difficult (II). (United States)

    Goodwin, Alan


    Presents some questions that science teachers find difficult. Focuses on three further questions relating to "simple" everyday situations that are normally explained in terms of the kinetic theory of matter. Identifies looking at the difference between chemical and physical changes as the most problematic question. (Author/YDS)

  5. Teaching Students to Form Effective Questions (United States)

    Stafford, Tish


    The ability to question lies at the heart of human curiosity and is a necessary component of cognition. The author stresses that forming questions is essential to human thought and communication. As such, forming questions is a foundational process that cuts across curricular areas and is embedded in content standards across the nation, including…

  6. Question Answering for Dutch: Simple does it

    NARCIS (Netherlands)

    Hoekstra, A.H.; Hiemstra, Djoerd; van der Vet, P.E.; Huibers, Theo W.C.


    When people pose questions in natural language to search for information on the web, the role of question answering (QA) systems becomes important. In this paper the QAsystem simpleQA, capable of answering Dutch questions on which the answer is a person or a location, is described. The system's

  7. Answering Wh- Questions About Sentences and Text. (United States)

    Singer, Murray


    Describes a study designed to identify the mental operations that contribute to people's ability to answer wh- questions, that is, questions which request information that plays a particular role in relation to some action or event. Wh- questions are signaled by interrogative pronouns and adverbs like who, what, when, and where. (SED)

  8. Twenty Questions Games Always End With Yes

    CERN Document Server

    Gill, John T


    Huffman coding is often presented as the optimal solution to Twenty Questions. However, a caveat is that Twenty Questions games always end with a reply of "Yes," whereas Huffman codewords need not obey this constraint. We bring resolution to this issue, and prove that the average number of questions still lies between H(X) and H(X)+1.

  9. Teaching Culture: Questioning Perspectives on Our Practices (United States)

    Met, Myriam


    Despite years of training, teaching experience, reading professional literature, attending conferences, and learning from expert colleagues, when it comes to the teaching of culture, the author wishes she knew more answers to many critical questions. Her questions are framed by the basic questions that all curricula seek to answer: WHAT is the…

  10. Questions That Science Teachers Find Difficult (II). (United States)

    Goodwin, Alan


    Presents some questions that science teachers find difficult. Focuses on three further questions relating to "simple" everyday situations that are normally explained in terms of the kinetic theory of matter. Identifies looking at the difference between chemical and physical changes as the most problematic question. (Author/YDS)

  11. Structural insights into electron transfer in caa 3-type cytochrome oxidase


    Lyons, Joseph A.; Aragão, David; Slattery, Orla; Pisliakov, Andrei V.; Soulimane, Tewfik; Caffrey, Martin


    Summary Paragraph Cytochrome c oxidase is a member of the heme copper oxidase superfamily (HCO) 1 . HCOs function as the terminal enzymes in the respiratory chain of mitochondria and aerobic prokaryotes, coupling molecular oxygen reduction to transmembrane proton pumping. Integral to the enzyme’s function is the transfer of electrons from cytochrome c to the oxidase via a transient association of the two proteins. Electron entry and exit are proposed to occur from the same site on cytochrome ...

  12. All About Alternatives (United States)

    Barr, Robert D.; And Others


    A primer on alternative schools. Described are existing programs in different areas, philosophy of the alternative schools, funding, student behavior, community relations, accountability, State regulations, management, and the environment of the alternative school. A list of sources of additional information on alternative schools is included.…

  13. Automatic Question Answering from Web Documents

    Institute of Scientific and Technical Information of China (English)

    LI Xin; HU Dawei; LI Huan; HAO Tianyong; CHEN Enhong; LIU Wenyin


    A passage retrieval strategy for web-based question answering (QA) systems is proposed in our QA system. It firstly analyzes the question based on semantic patterns to obtain its syntactic and semantic information and then form initial queries. The queries are used to retrieve documents from the World Wide Web (WWW) using the Google search engine. The queries are then rewritten to form queries for passage retrieval in order to improve the precision. The relations between keywords in the question are employed in our query rewrite method. The experimental result on the question set of the TREC-2003 passage task shows that our system performs well for factoid questions.

  14. Art of Question in English Teaching

    Institute of Scientific and Technical Information of China (English)



    A language is the tool of human communication. English teaching is really not a way to input or output information-it is not an one-way transmission of information. Students must personally think, speak and write what they are taught. That is the practical features of English teaching. Questioning is one of important methods which embody this feature completely. Teachers apply asking questions to enlighten and stimulate students in order to perfect this vital importance. Students who are in middle schools are asked questions but do not give answers. Except for learners' own quality and degree of difficulty of questions, it is connected with teachers' quality of asking questions.

  15. Unsubstituted phenothiazine as a superior water-insoluble mediator for oxidases


    Sekretaryova, Alina; Vagin, Mikhail; Beni, Valerio; Turner, Anthony P.F.; Karyakin, Arkady A


    The mediation of oxidases glucose oxidase (GOx), lactate oxidase (LOx) and cholesterol oxidase (ChOx) by a new electron shuttling mediator, unsubstituted phenothiazine (PTZ), was studied. Cyclic voltammetry and rotating-disk electrode measurements in nonaqueous media were used to determine the diffusion characteristics of the mediator and the kinetics of its reaction with GOx, giving a second-order rate constant of 7.6×103–2.1×104 M−1 s−1 for water–acetonitrile solutions containing 5–15% wate...

  16. Unanswered Questions in Contemporary Heart Failure. (United States)

    Gilstrap, Lauren G; Snipelisky, David; AbouEzzeddine, Omar; Vader, Justin; Cooper, Lauren; Kelley, Jacob; Perez, Antonio; Varian, Kenneth; Lala, Anuradha; Shah, Monica; Stevenson, Lynne W


    The epidemiology of heart failure (HF) is changing. This study aimed to describe questions that arise during the routine care of HF patients that are unanswered by the current literature and describe how the type and focus of these questions has changed over time. Investigators from the National Heart, Lung, and Blood Institute-sponsored Heart Failure Apprentice Network collected and categorized questions from 5 academic hospitals over 12 months. A total of 174 unanswered questions were collected and analyzed. Compared with 2004, there were more unanswered questions about "whether" to use therapies and fewer about "how" to use therapies. There were fewer questions about what therapeutic targets, therapy adjustment, and combination therapies. There were more questions about whether or how to stop therapies and how to add therapies back. Newly prominent topics, not observed in 2004, including novel therapeutics, refractory ventricular tachycardia, right heart failure, and nutrition/frailty, accounted for 24% of questions. Compared with 2004, there are fewer unanswered questions about how to use, adjust, and combine therapies. There were more unanswered questions about whether and how to stop therapies. Almost 25% of unanswered questions dealt with topics indicative of more advanced disease which were not observed in 2004. Copyright © 2017 Elsevier Inc. All rights reserved.

  17. Generalization and Alternatives of Kaprekar's Routine

    CERN Document Server

    Smarandache, Florentin


    We extend Kaprekar's Routine for a large class of applications. We also give particular examples of this generalization as alternatives to Kaprekar's Routine and Number. Some open questions about the length of the iterations until reaching either zero or a constant or a cycle, and about the length of the cycles are asked at the end.

  18. Alternative, complementary and traditional medicine in Malaysia. (United States)

    Talib, N


    This paper sets out the practice of traditional, alternative and/or complementary medicine in Malaysia. It gives an overview of the types of alternative medicine available, and the legal regulation, or lack of it within the current setting. The relevant policies and governmental action in this area are highlighted. Relevant case law decisions in this area are also included. The practice of spiritual healing as one form of traditional medicine, and its role within the spectrum of alternative medicine is dealt with briefly. The significant question of integration of alternative medicine within the existing allopathic system is addressed. The paper concludes that as interest in, and usage of alternative medicine is not likely to decrease, certain measures must be taken by the relevant authorities to ensure among others, the safety and efficacy of these medicines.

  19. Pretreatment with mixed-function oxidase inducers increases the sensitivity of the hepatocyte/DNA repair assay

    Energy Technology Data Exchange (ETDEWEB)

    Shaddock, J.G.; Heflich, R.H.; McMillan, D.C.; Hinson, J.A.; Casciano, D.A. (National Center for Toxicological Research, Jefferson, AK (USA) Univ. of Arkansas for Medical Sciences, Little Rock (USA))


    A recent National Toxicology Program evaluation indicates that the rat hepatocyte/DNA repair assay has a high false-negative rate and that it is insensitive to some genotoxic hepatocarcinogens as well as other species and organ-specific carcinogens. In this study, the authors examined whether the sensitivity of the hepatocyte/DNA repair assay might be increased through animal pretreatment with various hepatic mixed-function oxidase inducers, i.e., Aroclor 1254, phenobarbital, and 3,3{prime},4,4{prime}-tetrachloroazobenzene (TCAB). The effects on unscheduled DNA synthesis (UDS), a measured of DNA damage and repair, were studied in cultures exposed to known and/or potential carcinogens that had been evaluated as negative or questionable or that produced conflicting results with hepatocytes isolated from uninduced animals. 4,4{prime}-Oxydianiline, 1-nitropy-rene, and TCAB produced concentration-dependent increases in UDS in hepatocytes from rats pretreated with Aroclor 1254. 4,4{prime}-Oxydianiline and TCAB also induced a dose-dependent increase in DNA repair in hepatocytes from rats pretreated with phenobarbital, whereas 1-nitropyrene was negative. These data indicate that the limited sensitivity to chemical carcinogens displayed by the hepatocyte/DNA repair assay may be increased by using hepatocytes isolated from animals exposed to hepatic mixed-function oxidase inducers.

  20. Pyruvate oxidase is a determinant of Avery's rough morphology. (United States)

    Belanger, Aimee E; Clague, Melissa J; Glass, John I; Leblanc, Donald J


    In pioneering studies, Avery et al. identified DNA as the hereditary material (A. T. Avery, C. M. MacLeod, and M. McCarty, J. Exp. Med. 79:137-158, 1944). They demonstrated, by means of variation in colony morphology, that this substance could transform their rough type 2 Streptococcus pneumoniae strain R36A into a smooth type 3 strain. It has become accepted as fact, from modern textbook accounts of these experiments, that smooth pneumococci make capsule, while rough strains do not. We found that rough-to-smooth morphology conversion did not occur in rough strains R36A and R6 when the ability to synthesize native type 2 capsule was restored. The continued rough morphology of these encapsulated strains was attributed to a second, since-forgotten, morphology-affecting mutation that was sustained by R36A during strain development. We used a new genome-PCR-based approach to identify spxB, the gene encoding pyruvate oxidase, as the mutated locus in R36A and R6 that, with unencapsulation, gives rise to rough colony morphology, as we know it. The variant spxB allele of R36A and R6 is associated with increased cellular pyruvate oxidase activity relative to the ancestral strain D39. Increased pyruvate oxidase activity alters colony shape by mediating cell death. R36A requires a wild-type spxB allele for the expression of smooth type 2 morphology but not for the expression of smooth type 3 morphology, the phenotype monitored by Avery et al. Thus, the mutated spxB allele did not impact their use of smooth morphology to identify the transforming principle.

  1. The HIV-1 Nef protein and phagocyte NADPH oxidase activation

    DEFF Research Database (Denmark)

    Vilhardt, Frederik; Plastre, Olivier; Sawada, Makoto;


    -regulation of phagocyte NADPH oxidase subunits. Nef mutants lacking motifs involved in the interaction with Vav and PAK failed to reproduce the effects of wild type Nef, suggesting a role for the Vav/Rac/PAK signaling pathway. The following results suggest a key role for Rac in the priming effect of Nef. (i) Inactivation...... of Rac by Clostridium difficile toxin B abolished the Nef effect. (ii) The fraction of activated Rac1 was increased in Nef-transduced cells, and (iii) the dominant positive Rac1(V12) mutant mimicked the effect of Nef. These results are to our knowledge the first analysis of the effect of Rac activation...

  2. Bioactive compounds of inhibiting xanthine oxidase from Selaginella labordei. (United States)

    Tan, Wen-Jie; Xu, Jia-Cheng; Li, Li; Chen, Ke-Li


    Four flavone compounds were isolated from the effective fractions inhibiting xanthine oxidase (XOD) of the medicinal plant Selaginella labordei with anti-virus activity, and the structures were elucidated as 4'-methylether robustaflavone (1), robustaflavone (2), eriodictyol (3) and amentoflavone (4). The 50% inhibitory concentration (IC(50)) of the three compounds of inhibiting XOD were 61.0, 0.199, 16.0 and 32.0 mg L(-1), respectively. All of these compounds were isolated from the species for the first time, and eriodictyol was found from Selaginellaceae for the first time. Among these compounds, robustaflavone has been reported as an effective compound against the hepatitis B virus.

  3. Dynamic Question Ordering in Online Surveys

    Directory of Open Access Journals (Sweden)

    Early Kirstin


    Full Text Available Online surveys have the potential to support adaptive questions, where later questions depend on earlier responses. Past work has taken a rule-based approach, uniformly across all respondents. We envision a richer interpretation of adaptive questions, which we call Dynamic Question Ordering (DQO, where question order is personalized. Such an approach could increase engagement, and therefore response rate, as well as imputation quality. We present a DQO framework to improve survey completion and imputation. In the general survey-taking setting, we want to maximize survey completion, and so we focus on ordering questions to engage the respondent and collect hopefully all information, or at least the information that most characterizes the respondent, for accurate imputations. In another scenario, our goal is to provide a personalized prediction. Since it is possible to give reasonable predictions with only a subset of questions, we are not concerned with motivating users to answer all questions. Instead, we want to order questions to get information that reduces prediction uncertainty, while not being too burdensome. We illustrate this framework with two case studies, for the prediction and survey-taking settings. We also discuss DQO for national surveys and consider connections between our statistics-based question-ordering approach and cognitive survey methodology.

  4. 150 Student Questions on Solar Physics (United States)

    Lopez, R. E.; Gross, N. A.; Knipp, D. J.


    The Center for Integrated Space Weather Modeling (CISM) holds a two-week Space Weather Summer School for introductory graduate students and space weather professionals to gain a system level understanding of the space environment and the effects of space weather. A typical day in the summer school consists of three morning lectures followed by an afternoon lab session. After the morning lectures, the participants are each asked to submit a question about the mornings topics on a question card. The lecturers then take the time to answer these questions prior to afternoon sessions. In the last 5 years over 1000 such question cards have been collected and cataloged. Despite detailed lectures by experts similar questions appear every year. We have analyzed over 150 questions related to the introductory lectures on solar physics and solar activity. Questions content was categorized using the AGU Index, and question sophistication was categorized using Bloom’s Taxonomy of Educational Objectives. Specific analysis results along with lists of questions will be presented. We hope that these results can be used to improve the lecture and classroom content and allow students to move beyond low level education objectives and ask more sophisticated questions.

  5. Assessing the Psychometric Properties of Alternative Items for Certification. (United States)

    Krogh, Mary Anne; Muckle, Timothy

    Alternative items were added as scored items to the National Certification Examination for Nurse Anesthetists (NCE) in 2010. A common concern related to the new items has been their measurement attributes. This study was undertaken to evaluate the psychometric impact of adding these items to the examination. Candidates had a significantly higher ability estimate in alternative items than in multiple choice questions and 6.7 percent of test candidates performed significantly differently in alternative item formats. The ability estimates of multiple choice questions correlated at r = .58. The alternative items took significantly longer time to answer than standard multiple choice questions and discriminated to a higher degree than MCQs. The alternative items exhibited unidimensionality to the same degree as MCQs and the BIC confirmed the Rasch model as acceptable for scoring. The new item types were found to have acceptable attributes for inclusion in the certification program.

  6. Glutamic acid 242 is a valve in the proton pump of cytochrome c oxidase (United States)

    Kaila, Ville R. I.; Verkhovsky, Michael I.; Hummer, Gerhard; Wikström, Mårten


    Aerobic life is based on a molecular machinery that utilizes oxygen as a terminal electron sink. The membrane-bound cytochrome c oxidase (CcO) catalyzes the reduction of oxygen to water in mitochondria and many bacteria. The energy released in this reaction is conserved by pumping protons across the mitochondrial or bacterial membrane, creating an electrochemical proton gradient that drives production of ATP. A crucial question is how the protons pumped by CcO are prevented from flowing backwards during the process. Here, we show by molecular dynamics simulations that the conserved glutamic acid 242 near the active site of CcO undergoes a protonation state-dependent conformational change, which provides a valve in the pumping mechanism. The valve ensures that at any point in time, the proton pathway across the membrane is effectively discontinuous, thereby preventing thermodynamically favorable proton back-leakage while maintaining an overall high efficiency of proton translocation. Suppression of proton leakage is particularly important in mitochondria under physiological conditions, where production of ATP takes place in the presence of a high electrochemical proton gradient. PMID:18430799

  7. Question Classification Taxonomies as Guides to Formulating Questions for Use in Chemistry Classrooms (United States)

    Festo, Kayima


    Teacher questions play an important role in facilitating classroom discourse. Using appropriate question types and proper questioning techniques help to create reflective-active learners. Teacher questions can elicit students' explanations, elaboration of their ideas and thinking, and they can be used to disclose students' misconceptions. Despite…

  8. Structured Attentions for Visual Question Answering


    Zhu, Chen; Zhao, Yanpeng; Huang, Shuaiyi; Tu, Kewei; Ma, Yi


    Visual attention, which assigns weights to image regions according to their relevance to a question, is considered as an indispensable part by most Visual Question Answering models. Although the questions may involve complex relations among multiple regions, few attention models can effectively encode such cross-region relations. In this paper, we demonstrate the importance of encoding such relations by showing the limited effective receptive field of ResNet on two datasets, and propose to mo...

  9. On the intonation of German intonation questions

    DEFF Research Database (Denmark)

    Petrone, Caterina; Niebuhr, Oliver


    German questions and statements are distinguished not only by lexical and syntactic but also by intonational means. This study revisits, for Northern Standard German, how questions are signalled intonationally in utterances that have neither lexical nor syntactic cues. Starting from natural...... productions of such 'intonation questions', two perception experiments were run. Experiment 1 is based on a gating paradigm, which was applied to naturally produced questions and statements. Experiment 11 includes two indirect-identification tasks. Resynthesized stimuli were judged in relation to two context...

  10. Partial purification and characterization of polyphenol oxidase from persimmon. (United States)

    Navarro, José L; Tárrega, Amparo; Sentandreu, Miguel A; Sentandreu, Enrique


    Activity of polyphenol oxidase (PPO) from "Rojo Brillante" persimmon (Diospyros kaki L.) fruits was characterized. Crude extracts were used for characterization of enzyme activity and stability at different temperatures (60, 70 and 80 °C), pHs (from 3.5 to 7.5) and substrate concentrations (catechol from 0 to 0.5M). Maximum enzyme activity was reached at pH 5.5 and 55 °C. Enzyme stability was higher than PPO activities found in other natural sources, since above pH 5.5 the minimum time needed to achieve an enzyme inactivation of 90% was 70 min at 80 °C. However, at pH 4.0 the enzyme stability decreased, reaching inactivation levels above 90% after 10 min even at 60 °C. Thus it was concluded that acidification can circumvent browning problems caused by PPO activity. Moreover, polyacrylamide gel electrophoresis of the enriched extract revealed the presence of at least four bands with strong oxidase activity, suggesting the existence of different PPO isoforms.

  11. Differential Expression of the Three Multicopper Oxidases from Myxococcus xanthus▿ (United States)

    Sánchez-Sutil, María Celestina; Gómez-Santos, Nuria; Moraleda-Muñoz, Aurelio; Martins, Lígia O.; Pérez, Juana; Muñoz-Dorado, José


    Myxococcus xanthus is a soil bacterium that undergoes a unique life cycle among the prokaryotes upon starvation, which includes the formation of macroscopic structures, the fruiting bodies, and the differentiation of vegetative rods into coccoid myxospores. This peculiarity offers the opportunity to study the copper response in this bacterium in two different stages. In fact, M. xanthus vegetative rods exhibit 15-fold-greater resistance against copper than developing cells. However, cells preadapted to this metal reach the same levels of resistance during both stages. Analysis of the M. xanthus genome reveals that many of the genes involved in copper resistance are redundant, three of which encode proteins of the multicopper oxidase family (MCO). Each MCO gene exhibits a different expression profile in response to external copper addition. Promoters of cuoA and cuoB respond to Cu(II) ions during growth and development; however, they show a 10-fold-increased copper sensitivity during development. The promoter of cuoC shows copper-independent induction upon starvation, but it is copper up-regulated during growth. Phenotypic analyses of deletion mutants reveal that CuoB is involved in the primary copper-adaptive response; CuoA and CuoC are necessary for the maintenance of copper tolerance; and CuoC is required for normal development. These roles seem to be carried out through cuprous oxidase activity. PMID:17483223

  12. Crystal Structure of the Deglycating Enzyme Fructosamine Oxidase (Amadoriase ll)

    Energy Technology Data Exchange (ETDEWEB)

    Collard, F.; Zhang, J; Nemet, I; Qanungo, K; Monnier, V; Yee, V


    Fructosamine oxidases (FAOX) catalyze the oxidative deglycation of low molecular weight fructosamines (Amadori products). These proteins are of interest in developing an enzyme to deglycate proteins implicated in diabetic complications. We report here the crystal structures of FAOX-II from the fungi Aspergillus fumigatus, in free form and in complex with the inhibitor fructosyl-thioacetate, at 1.75 and 1.6{angstrom} resolution, respectively. FAOX-II is a two domain FAD-enzyme with an overall topology that is most similar to that of monomeric sarcosine oxidase. Active site residues Tyr-60, Arg-112 and Lys-368 bind the carboxylic portion of the fructosamine, whereas Glu-280 and Arg-411 bind the fructosyl portion. From structure-guided sequence comparison, Glu-280 was identified as a signature residue for FAOX activity. Two flexible surface loops become ordered upon binding of the inhibitor in a catalytic site that is about 12{angstrom} deep, providing an explanation for the very low activity of FAOX enzymes toward protein-bound fructosamines, which would have difficulty accessing the active site. Structure-based mutagenesis showed that substitution of Glu-280 and Arg-411 eliminates enzyme activity. In contrast, modification of other active site residues or of amino acids in the flexible active site loops has little effect, highlighting these regions as potential targets in designing an enzyme that will accept larger substrates.

  13. Crystal Structure of the Deglycating Enzyme Fructosamine Oxidase (Amadoriase II)

    Energy Technology Data Exchange (ETDEWEB)

    Collard, François; Zhang, Jianye; Nemet, Ina; Qanungo, Kaustubha R.; Monnier, Vincent M.; Yee, Vivien C. (Case Western)


    Fructosamine oxidases (FAOX) catalyze the oxidative deglycation of low molecular weight fructosamines (Amadori products). These proteins are of interest in developing an enzyme to deglycate proteins implicated in diabetic complications. We report here the crystal structures of FAOX-II from the fungi Aspergillus fumigatus, in free form and in complex with the inhibitor fructosyl-thioacetate, at 1.75 and 1.6{angstrom} resolution, respectively. FAOX-II is a two domain FAD-enzyme with an overall topology that is most similar to that of monomeric sarcosine oxidase. Active site residues Tyr-60, Arg-112 and Lys-368 bind the carboxylic portion of the fructosamine, whereas Glu-280 and Arg-411 bind the fructosyl portion. From structure-guided sequence comparison, Glu-280 was identified as a signature residue for FAOX activity. Two flexible surface loops become ordered upon binding of the inhibitor in a catalytic site that is about 12{angstrom} deep, providing an explanation for the very low activity of FAOX enzymes toward protein-bound fructosamines, which would have difficulty accessing the active site. Structure-based mutagenesis showed that substitution of Glu-280 and Arg-411 eliminates enzyme activity. In contrast, modification of other active site residues or of amino acids in the flexible active site loops has little effect, highlighting these regions as potential targets in designing an enzyme that will accept larger substrates.

  14. Crystal Structure of the Deglycating Enzyme Fructosamine Oxidase (Amadoriase II)* (United States)

    Collard, François; Zhang, Jianye; Nemet, Ina; Qanungo, Kaustubha R.; Monnier, Vincent M.; Yee, Vivien C.


    Fructosamine oxidases (FAOX) catalyze the oxidative deglycation of low molecular weight fructosamines (Amadori products). These proteins are of interest in developing an enzyme to deglycate proteins implicated in diabetic complications. We report here the crystal structures of FAOX-II from the fungi Aspergillus fumigatus, in free form and in complex with the inhibitor fructosyl-thioacetate, at 1.75 and 1.6Å resolution, respectively. FAOX-II is a two domain FAD-enzyme with an overall topology that is most similar to that of monomeric sarcosine oxidase. Active site residues Tyr-60, Arg-112 and Lys-368 bind the carboxylic portion of the fructosamine, whereas Glu-280 and Arg-411 bind the fructosyl portion. From structure-guided sequence comparison, Glu-280 was identified as a signature residue for FAOX activity. Two flexible surface loops become ordered upon binding of the inhibitor in a catalytic site that is about 12Å deep, providing an explanation for the very low activity of FAOX enzymes toward protein-bound fructosamines, which would have difficulty accessing the active site. Structure-based mutagenesis showed that substitution of Glu-280 and Arg-411 eliminates enzyme activity. In contrast, modification of other active site residues or of amino acids in the flexible active site loops has little effect, highlighting these regions as potential targets in designing an enzyme that will accept larger substrates. PMID:18667417

  15. Crystal structure of the deglycating enzyme fructosamine oxidase (amadoriase II). (United States)

    Collard, François; Zhang, Jianye; Nemet, Ina; Qanungo, Kaustubha R; Monnier, Vincent M; Yee, Vivien C


    Fructosamine oxidases (FAOX) catalyze the oxidative deglycation of low molecular weight fructosamines (Amadori products). These proteins are of interest in developing an enzyme to deglycate proteins implicated in diabetic complications. We report here the crystal structures of FAOX-II from the fungi Aspergillus fumigatus, in free form and in complex with the inhibitor fructosyl-thioacetate, at 1.75 and 1.6A resolution, respectively. FAOX-II is a two domain FAD-enzyme with an overall topology that is most similar to that of monomeric sarcosine oxidase. Active site residues Tyr-60, Arg-112 and Lys-368 bind the carboxylic portion of the fructosamine, whereas Glu-280 and Arg-411 bind the fructosyl portion. From structure-guided sequence comparison, Glu-280 was identified as a signature residue for FAOX activity. Two flexible surface loops become ordered upon binding of the inhibitor in a catalytic site that is about 12A deep, providing an explanation for the very low activity of FAOX enzymes toward protein-bound fructosamines, which would have difficulty accessing the active site. Structure-based mutagenesis showed that substitution of Glu-280 and Arg-411 eliminates enzyme activity. In contrast, modification of other active site residues or of amino acids in the flexible active site loops has little effect, highlighting these regions as potential targets in designing an enzyme that will accept larger substrates.

  16. 2-acetylphenol analogs as potent reversible monoamine oxidase inhibitors

    Directory of Open Access Journals (Sweden)

    Legoabe LJ


    Full Text Available Lesetja J Legoabe,1 Anél Petzer,1 Jacobus P Petzer1,21Centre of Excellence for Pharmaceutical Sciences, 2Department of Pharmaceutical Chemistry, School of Pharmacy, North-West University, Potchefstroom, South AfricaAbstract: Based on a previous report that substituted 2-acetylphenols may be promising leads for the design of novel monoamine oxidase (MAO inhibitors, a series of C5-substituted 2-acetylphenol analogs (15 and related compounds (two were synthesized and evaluated as inhibitors of human MAO-A and MAO-B. Generally, the study compounds exhibited inhibitory activities against both MAO-A and MAO-B, with selectivity for the B isoform. Among the compounds evaluated, seven compounds exhibited IC50 values <0.01 µM for MAO-B inhibition, with the most selective compound being 17,000-fold selective for MAO-B over the MAO-A isoform. Analyses of the structure–activity relationships for MAO inhibition show that substitution on the C5 position of the 2-acetylphenol moiety is a requirement for MAO-B inhibition, and the benzyloxy substituent is particularly favorable in this regard. This study concludes that C5-substituted 2-acetylphenol analogs are potent and selective MAO-B inhibitors, appropriate for the design of therapies for neurodegenerative disorders such as Parkinson’s disease.Keywords: monoamine oxidase, MAO, inhibition, 2-acetylphenol, structure–activity relationship

  17. Inhibition of Xanthine Oxidase Activity by Gnaphalium Affine Extract

    Institute of Scientific and Technical Information of China (English)

    Wei-qing Lin; Jian-xiang Xie; Xiao-mu Wu; Lin Yang; Hai-dong Wang


    Objective To evaluate the inhibitory effect of Gnaphalium affine extracts on xanthine oxidase (XO) activity in vitro and to analyze the mechanism of this effect. Methods In this in vitro study, Kinetic measurements were performed in 4 different inhibitor concentrations and 5 different xanthine concentrations (60, 100, 200, 300, 400 μmol/L). Dixon and Lineweaver-Burk plot analysis were used to determine Ki values and the inhibition mode for the compounds isolated from Gnaphalium affine extract. Results Four potent xanthine oxidase inhibitors were found in 95% ethanolic (v/v) Gnaphalium affine extract. Among them, the flavone Eupatilin exhibited the strongest inhibitory effect on XO with a inhibition constant (Ki) of 0.37μmol/L, lower than the Ki of allopurinol (4.56 mol/L), a known synthetic XO inhibitor. Apigenin (Ki of 0.56μmol/L, a proportion of 0.0053‰in Gnaphalium affine), luteolin (Ki of 2.63 μmol/L, 0.0032‰ in Gnaphalium affine) and 5-hydroxy-6,7,3’,4’-tetramethoxyflavone (Ki of 3.15μmol/L, 0.0043‰ in Gnaphalium affine) also contributed to the inhibitory effect of Gnaphalium affine extract on XO activity. Conclusions These results suggest that the use of Gnaphalium affine in the treatment of gout could be attributed to its inhibitory effect on XO. This study provides a rational basis for the traditional use of Gnaphalium affine against gout.

  18. ADP competes with FAD binding in putrescine oxidase. (United States)

    van Hellemond, Erik W; Mazon, Hortense; Heck, Albert J; van den Heuvel, Robert H H; Heuts, Dominic P H M; Janssen, Dick B; Fraaije, Marco W


    Putrescine oxidase from Rhodococcus erythropolis NCIMB 11540 (PuO(Rh)) is a soluble homodimeric flavoprotein of 100 kDa, which catalyzes the oxidative deamination of putrescine and some other aliphatic amines. The initial characterization of PuO(Rh) uncovered an intriguing feature: the enzyme appeared to contain only one noncovalently bound FAD cofactor per dimer. Here we show that this low FAD/protein ratio is the result of tight binding of ADP, thereby competing with FAD binding. MS analysis revealed that the enzyme is isolated as a mixture of dimers containing two molecules of FAD, two molecules ADP, or one FAD and one ADP molecule. In addition, based on a structural model of PuO(Rh) that was built using the crystal structure of human monoamine oxidase B (MAO-B), we constructed an active mutant enzyme, PuO(Rh) A394C, that contains covalently bound FAD. These findings show that the covalent FAD-protein linkage can be formed autocatalytically and hint to a new-found rationale for covalent flavinylation: covalent flavinylation may have evolved to prevent binding of ADP or related cellular compounds, which would prohibit formation of flavinylated and functional enzyme.

  19. Vowel alternations in English


    Kazumi, Yukiko


    The purpose of this study is to investigate the nature of vowel alternations observed in English. What we call vowel alternations here consists of shortening and lengthening triggered by Level I affixation: ...

  20. The Oxidation of Thiols by Flavoprotein Oxidases : a Biocatalytic Route to Reactive Thiocarbonyls

    NARCIS (Netherlands)

    Ewing, Tom A.; Dijkman, Willem P.; Vervoort, Jacques M.; Fraaije, Marco W.; van Berkel, Willem J. H.


    Flavoprotein oxidases are a diverse class of biocatalysts, most of which catalyze the oxidation of C-O, C-N, or C-C bonds. Flavoprotein oxidases that are known to catalyze the oxidation of C-S bonds are rare, being limited to enzymes that catalyze the oxidative cleavage of thioethers. Herein, we rep

  1. The Oxidation of Thiols by Flavoprotein Oxidases: a Biocatalytic Route to Reactive Thiocarbonyls.

    NARCIS (Netherlands)

    Ewing, T.A.; Dijkman, W.P.; Vervoort, J.J.M.; Fraaije, M.W.; Berkel, van W.J.H.


    Flavoprotein oxidases are a diverse class of biocatalysts, most of which catalyze the oxidation of C[BOND]O, C[BOND]N, or C[BOND]C bonds. Flavoprotein oxidases that are known to catalyze the oxidation of C[BOND]S bonds are rare, being limited to enzymes that catalyze the oxidative cleavage of thioet

  2. Azide binding to the trinuclear copper center in laccase and ascorbate oxidase

    DEFF Research Database (Denmark)

    Gromov, I; Marchesini, A; Farver, O


    Azide binding to the blue copper oxidases laccase and ascorbate oxidase (AO) was investigated by electron paramagnetic resonance (EPR) and pulsed electron-nuclear double resonance (ENDOR) spectroscopies. As the laccase : azide molar ratio decreases from 1:1 to 1:7, the intensity of the type 2 (T2...

  3. Hydroxychavicol: a potent xanthine oxidase inhibitor obtained from the leaves of betel, Piper betle. (United States)

    Murata, Kazuya; Nakao, Kikuyo; Hirata, Noriko; Namba, Kensuke; Nomi, Takao; Kitamura, Yoshihisa; Moriyama, Kenzo; Shintani, Takahiro; Iinuma, Munekazu; Matsuda, Hideaki


    The screening of Piperaceous plants for xanthine oxidase inhibitory activity revealed that the extract of the leaves of Piper betle possesses potent activity. Activity-guided purification led us to obtain hydroxychavicol as an active principle. Hydroxychavicol is a more potent xanthine oxidase inhibitor than allopurinol, which is clinically used for the treatment of hyperuricemia.

  4. Biocatalytic potential of laccase-like multicopper oxidases from Aspergillus niger

    NARCIS (Netherlands)

    Tamayo Ramos, J.A.; Berkel, van W.J.H.; Graaff, de L.H.


    BACKGROUND: Laccase-like multicopper oxidases have been reported in several Aspergillus species but they remain uncharacterized. The biocatalytic potential of the Aspergillus niger fungal pigment multicopper oxidases McoA and McoB and ascomycete laccase McoG was investigated. RESULTS: The laccase-li

  5. Analysis of cellulase and polyphenol oxidase production by southern pine beetle associated fungi (United States)

    Abduvali Valiev; Zumrut B. Ogel; Dier D. Klepzig


    In this study, the production of extracellular enzymes by fungi associated with southern pine beetle was investigated for the first time. Cellulase and polyphenol oxidase production were analyzed for three beetle associated fungi. Only the mutualistic symbiont Entomocorticium sp. A was found to produce cellulases and polyphenol oxidase....

  6. Purification, characterization and decolorization of bilirubin oxidase from Myrothecium verrucaria 3.2190 (United States)

    Myrothecium verrucaria 3.2190 is a nonligninolytic fungus that produces bilirubin oxidase. Both Myrothecium verrucaria and the extracellular bilirubin oxidase were tested for their ability to decolorize indigo carmine. The biosorption and biodegradation of the dye were detected during the process of...

  7. Process technology for the application of d-amino acid oxidases in pharmaceutical intermediate manufacturing

    DEFF Research Database (Denmark)

    Tindal, Stuart; Carr, Reuben; Archer, Ian V. J.


    Recent advances in biocatalysis have seen increased interest in the use of D-amino acid oxidase to synthesize optically pure amino acids. However, the creation of a genuine oxidase based platform technology will require suitable process technology as well as an understanding of the challenges and...

  8. Effects of the NADPH oxidase inhibitor apocynin on the left ventricular dysfunction induced by cocaine administration

    Institute of Scientific and Technical Information of China (English)

    MarcISABELLE; ChristelleMONTEIL; ChristianTHUILLEZ


    AIM: In a previous study, we have shown the role of alphaladrenoceptor in the left ventricular (LV) dysfunction after chronic cocaine administration via the induction of NADPH oxidase. In this study we used the NADPH oxidase inhibitor apocynin, to further investigate the real involvement of this prooxidant system in this LV dysfunction. METHODS: Wistar rats were treated

  9. Structural analysis of the catalytic mechanism and stereo selectivity in Streptomyces coelicolor alditol oxidase

    NARCIS (Netherlands)

    Forneris, Federico; Heuts, Dominic P. H. M.; Delvecchio, Manuela; Rovida, Stefano; Fraaije, Marco W.; Mattevi, Andrea


    Alditol oxidase (AldO) from Streptomyces coelicolor A3(2) is a soluble monomeric flavin-dependent oxidase that performs selective oxidation of the terminal primary hydroxyl group of several alditols. Here, we report the crystal structure of the recombinant enzyme in its native state and in complex w

  10. Spectral and catalytic properties of aryl-alcohol oxidase, a fungal flavoenzyme acting on polyunsaturated alcohols

    NARCIS (Netherlands)

    Ferreira, P.; Medina, M.; Guillén, F.; Martínez, M.J.; Berkel, van W.J.H.; Martínez, A.T.


    Spectral and catalytic properties of the flavoenzyme AAO (aryl-alcohol oxidase) from Pleurotus eryngii were investigated using recombinant enzyme. Unlike most flavoprotein oxidases, AAO does not thermodynamically stabilize a flavin semiquinone radical and forms no sulphite adduct. AAO catalyses the

  11. Biodegradation of phenolic compounds with oxidases from sorghum and non-defined mixed bacterium media

    Energy Technology Data Exchange (ETDEWEB)

    Obame, C. E. L.; Savadogo, P. W.; Mamoudou, D. H.; Dembele, R. H.; Traore, A. S.


    The biodegradation of the phenolic compounds is performed using oxidative enzymes, e. g. polyphenol oxidases (PPOs) and peroxidases (POXs). These oxidases displaying a wide spectrum for the oxidation of phenolic compounds were isolated either from sorghum or mixed bacteria. Spectrophotometric methods were used to assess the monophenolase and diphenolase activities of PPOs as well as the hydrogen-dependant oxidation of POXs. (Author)

  12. Vectorial nature of redox Bohr effects in bovine heart cytochrome c oxidase. (United States)

    Capitanio, N; Capitanio, G; De Nitto, E; Papa, S


    The vectorial nature of redox Bohr effects (redox-linked pK shifts) in cytochrome c oxidase from bovine heart incorporated in liposomes has been analyzed. The Bohr effects linked to oxido-reduction of heme a and CuB display membrane vectorial asymmetry. This provides evidence for involvement of redox Bohr effects in the proton pump of the oxidase.


    NARCIS (Netherlands)



    Alcohol oxidase of methylotrophic yeast is an FAD-containing enzyme. When in its active form, the enzyme is an octamer and located in the peroxisomes. To study the importance of FAD-binding on the activity, octamerization and intracellular localization of the enzyme, alcohol oxidase of Hansenula pol

  14. New Markers: Urine Xanthine Oxidase and Myeloperoxidase in the Early Detection of Urinary Tract Infection

    Directory of Open Access Journals (Sweden)

    Pınar Ciragil


    Full Text Available Objectives. The aim of this study was to evaluate if xanthine oxidase and myeloperoxidase levels quantitation method may alternate routine culture method, which takes more time in the diagnosis of urinary tract infections. Material and Methods. Five hundred and forty-nine outpatients who had admitted to Clinic Microbiology Laboratory were included in the study. The microorganisms were identified by using VITEK System. The urine specimens that were negative from the quantitative urine culture were used as controls. The activities of MPO and XO in spot urine were measured by spectrophotometric method. Results. Through the urine cultures, 167 bacteria were isolated from 163 urine specimens; 386 cultures yielded no bacterial growth. E. coli was the most frequent pathogen. In infection with E. coli both XO and MPO levels were increased the most. The sensitivity, specificity, positive predictive value, and negative predictive value for XO were 100%, 100%, 100%, and 100%, respectively. These values for MPO were 87%, 100%, 100%, and 94%, respectively. Conclusion. These data obtained suggest that urine XO and MPO levels may be new markers in the early detection of UTI.

  15. High-level expression of the Penicillium notatum glucose oxidase gene in Pichia pastoris using codon optimization. (United States)

    Gao, Zhaowei; Li, Zhuofu; Zhang, Yuhong; Huang, Huoqing; Li, Mu; Zhou, Liwei; Tang, Yunming; Yao, Bin; Zhang, Wei


    The glucose oxidase (GOD) gene from Penicillium notatum was expressed in Pichia pastoris. The 1,815 bp gene, god-w, encodes 604 amino acids. Recombinant GOD-w had optimal activity at 35-40°C and pH 6.2 and was stable, from pH 3 to 7 maintaining >75% maximum activity after incubation at 50°C for 1 h. GOD-w worked as well as commercial GODs to improve bread making. To achieve high-level expression of recombinant GOD in P. pastoris, 272 nucleotides involving 228 residues were mutated, consistent with the codon bias of P. pastoris. The optimized recombinant GOD-m yielded 615 U ml(-1) (2.5 g protein l(-1)) in a 3 l fermentor--410% higher than GOD-w (148 U ml(-1)), and thus is a low-cost alternative for the bread baking industry.

  16. Rcf1 mediates cytochrome oxidase assembly and respirasome formation, revealing heterogeneity of the enzyme complex. (United States)

    Vukotic, Milena; Oeljeklaus, Silke; Wiese, Sebastian; Vögtle, F Nora; Meisinger, Chris; Meyer, Helmut E; Zieseniss, Anke; Katschinski, Doerthe M; Jans, Daniel C; Jakobs, Stefan; Warscheid, Bettina; Rehling, Peter; Deckers, Markus


    The terminal enzyme of the mitochondrial respiratory chain, cytochrome oxidase, transfers electrons to molecular oxygen, generating water. Within the inner mitochondrial membrane, cytochrome oxidase assembles into supercomplexes, together with other respiratory chain complexes, forming so-called respirasomes. Little is known about how these higher oligomeric structures are attained. Here we report on Rcf1 and Rcf2 as cytochrome oxidase subunits in S. cerevisiae. While Rcf2 is specific to yeast, Rcf1 is a conserved subunit with two human orthologs, RCF1a and RCF1b. Rcf1 is required for growth in hypoxia and complex assembly of subunits Cox13 and Rcf2, as well as for the oligomerization of a subclass of cytochrome oxidase complexes into respirasomes. Our analyses reveal that the cytochrome oxidase of mitochondria displays intrinsic heterogeneity with regard to its subunit composition and that distinct forms of respirasomes can be formed by complex variants.

  17. Cox26 is a novel stoichiometric subunit of the yeast cytochrome c oxidase. (United States)

    Levchenko, Maria; Wuttke, Jan-Moritz; Römpler, Katharina; Schmidt, Bernhard; Neifer, Klaus; Juris, Lisa; Wissel, Mirjam; Rehling, Peter; Deckers, Markus


    The cytochrome c oxidase (COX) is the terminal enzyme of the respiratory chain. The complex accepts electrons from cytochrome c and passes them onto molecular oxygen. This process contributes to energy capture in the form of a membrane potential across the inner membrane. The enzyme complex assembles in a stepwise process from the three mitochondria-encoded core subunits Cox1, Cox2 and Cox3, which associate with nuclear-encoded subunits and cofactors. In the yeast Saccharomyces cerevisiae, the cytochrome c oxidase associates with the bc1-complex into supercomplexes, allowing efficient energy transduction. Here we report on Cox26 as a protein found in respiratory chain supercomplexes containing cytochrome c oxidase. Our analyses reveal Cox26 as a novel stoichiometric structural subunit of the cytochrome c oxidase. A loss of Cox26 affects cytochrome c oxidase activity and respirasome organization.

  18. Retinal ganglion cells of high cytochrome oxidase activity in the rat

    Institute of Scientific and Technical Information of China (English)



    Retinal ganglion cells in the rat were studied using the heavy metal intensified cytochrome oxidase and horseradish peroxidase histochemical methods.The results show that a population of large retinal ganglion cells was consistently observed with the cytochrome oxidase staining method in retinas of normal rats or rats which received unilateral thalamotomy at birth.These cytochrome oxidase rich ganglion cells appeared to have large somata,3-6 primary dendrites and extensive dendritic arbors,and are comparable to ganglion cells labeled by the wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP).However,the morphological details of some of the cells revealed by the cytochrome oxidase staining method are frequently better than those shown by the HRP histochemical method.These results suggest that the mitochondrial enzyme cytochrome oxidase can be used as a simple but reliable marker for identifying and studying a population of retinal genglion cells with high metabolic rate in the rat.

  19. Children's questions: a mechanism for cognitive development. (United States)

    Chouinard, Michael M


    Preschoolers' questions may play an important role in cognitive development. When children encounter a problem with their current knowledge state (a gap in their knowledge, some ambiguity they do not know how to resolve, some inconsistency they have detected), asking a question allows them to get targeted information exactly when they need it. This information is available to them when they are particularly receptive to it, and because it comes as the result of their own disequilibrium, it may have depth of processing benefits. In that questions allow children to get information they need to move their knowledge structures closer to adult-like states, the ability to ask questions to gather needed information constitutes an efficient mechanism for cognitive development (referred to in this paper as the Information Requesting Mechanism [IRM]; this term is used because it includes question-asking and other information recruiting behaviors such as gestures, expressions, and vocalizations). However, the role of children's questions in their cognitive development has been largely overlooked. If questions are a force in cognitive development, the following must be true: (1) children must actually ask questions that gather information; (2) children must receive informative answers to their questions if they are able to be of use to cognitive development; (3) children must be motivated to get the information they request, rather than asking questions for other purposes such as attention; (4) the questions children ask must be relevant and of potential use to their cognitive development; (5) we must see evidence that children's questions help them in some way-that is, that they can ask questions for a purpose, and use the information they receive purposefully to successfully achieve some change of knowledge state. This monograph reports data on these points. Study 1 analyzed questions taken from four children's transcripts in the CHILDES database (age 1;2-5;1). This

  20. Spatiotemporal localization of D-amino acid oxidase and D-aspartate oxidases during development in Caenorhabditis elegans. (United States)

    Saitoh, Yasuaki; Katane, Masumi; Kawata, Tomonori; Maeda, Kazuhiro; Sekine, Masae; Furuchi, Takemitsu; Kobuna, Hiroyuki; Sakamoto, Taro; Inoue, Takao; Arai, Hiroyuki; Nakagawa, Yasuhito; Homma, Hiroshi


    Recent investigations have shown that a variety of D-amino acids are present in living organisms and that they possibly play important roles in physiological functions in the body. D-Amino acid oxidase (DAO) and D-aspartate oxidase (DDO) are degradative enzymes stereospecific for D-amino acids. They have been identified in various organisms, including mammals and the nematode Caenorhabditis elegans, although the significance of these enzymes and the relevant functions of D-amino acids remain to be elucidated. In this study, we investigated the spatiotemporal localization of C. elegans DAO and DDOs (DDO-1, DDO-2, and DDO-3) and measured the levels of several D- and L-amino acids in wild-type C. elegans and four mutants in which each gene for DAO and the DDOs was partially deleted and thereby inactivated. Furthermore, several phenotypes of these mutant strains were characterized. The results reported in this study indicate that C. elegans DAO and DDOs are involved in egg-laying events and the early development of C. elegans. In particular, DDOs appear to play important roles in the development and maturation of germ cells. This work provides novel and useful insights into the physiological functions of these enzymes and D-amino acids in multicellular organisms.

  1. Questions and dependency in intuitionistic logic

    NARCIS (Netherlands)

    Ciardelli, Ivano; Iemhoff, Rosalie; Yang, Fan


    In recent years, the logic of questions and dependencies has been investigated in the closely related frameworks of inquisitive logic and dependence logic. These investigations have assumed classical logic as the background logic of statements, and added formulas expressing questions and

  2. "Wh"-Questions in the English Language (United States)

    Mowarin, Macaulay


    This paper analyzes "wh"-questions in the English Language based mainly on Chomsky's Minimalist Programme of transformational grammar as the theoretical model. The four main objectives of this paper are as follows: first, it undertakes a cross linguistic typological analysis of "wh"-questions and it then discusses the derivation of…

  3. Fixed-Response Questions with a Difference. (United States)

    Johnstone, Alex H.; Ambusaidi, Abdullah


    Offers three types of fixed-response questions that are designed to overcome drawbacks appearing in the conventional forms of fixed-response questions such as not allowing the examiner to investigate reasoning, background, or prevent guessing. (Contains 14 references.) (Author/YDS)

  4. Teaching Dystopias: The Value of Religious Questioning. (United States)

    Seabury, Marcia Bundy


    Argues that a true general education should encourage the exploration of religious questions. Describes the author's use of works showing dystopian societies based on existing values, such as Huxley's "Brave New World," to encourage students to rethink their assumptions and develop openness toward the questions that religions address. (22…

  5. Questioning Techniques: A Study of Instructional Practice (United States)

    Hill, Joan Buchanan


    This study took place in an independent all girls' school which serves over six hundred students in grades pre-kindergarten through twelve. This study seeks to answer the question: To what extent do teachers use questions to encourage deeper thinking and fuller responses. Through a review of literature, observations, interviews and analysis,…

  6. Question Answering with Joost at CLEF 2007

    NARCIS (Netherlands)

    Bouma, G.; Kloosterman, G.; Mur, J.; van Noord, G.J.M.; van der Plas, M.L.E.; Tiedemann, J.; Peters, C.; Jijkoun, V.; Mandl, T.; Müller, H.; Oard, D.W.; Penas, A.; Petras, V.; Santos, D.


    We describe our system for the monolingual Dutch and multilingual English to Dutch QA tasks. We describe the preprocessing of Wikipedia, inclusion of query expansion in IR, anaphora resolution in follow-up questions, and a question classification module for the multilingual task. Our best runs achie

  7. Semantic Question Generation Using Artificial Immunity

    Directory of Open Access Journals (Sweden)

    Ibrahim E. Fattoh


    Full Text Available This research proposes an automatic question generation model for evaluating the understanding of semantic attributes in a sentence. The Semantic Role Labeling and Named Entity Recognition are used as a preprocessing step to convert the input sentence into a semantic pattern. The Artificial Immune System is used to build a classifier that will be able to classify the patterns according to the question type in the training phase. The question types considered here are the set of WH-questions like who, when, where, why, and how. A pattern matching phase is applied for selecting the best matching question pattern for the test sentence. The proposed model is tested against a set of sentences obtained from many sources such as the TREC 2007 dataset for question answering, Wikipedia articles, and English book of grade II preparatory. The experimental results of the proposed model are promising in determining the question type with classification accuracy reaching 95%, and 87% in generating the new question patterns.

  8. A Hybrid Approach to Clinical Question Answering (United States)


    paraphrasing and textual entailment) for better clinical reasoning and question answering. References O. Bodenreider. 2008. Biomedical Ontologies in...this track was to retrieve relevant biomedical articles to answer generic clini- cal questions about medical case reports. As part of our maiden...techniques to improve patient care through provid- ing pertinent biomedical information related to med- ical case reports. The primary motivation for

  9. "Wh"-Questions in the English Language (United States)

    Mowarin, Macaulay


    This paper analyzes "wh"-questions in the English Language based mainly on Chomsky's Minimalist Programme of transformational grammar as the theoretical model. The four main objectives of this paper are as follows: first, it undertakes a cross linguistic typological analysis of "wh"-questions and it then discusses the derivation of…

  10. The Notion of Coercion in Courtroom Questioning. (United States)

    Bulow-Moller, Anne Marie

    To account for coercive force in questions posed by counsel to defendants and witnesses, several levels of speech must be addressed. Forensic linguistics literature discusses the scale of coerciveness as reflected in the syntactic form of the questions. It is argued that this type of analysis fails to account for the inferences made by hearers…

  11. Mental Models of Research: Generating Authentic Questions (United States)

    Donham, Jean; Heinrich, Jill A.; Bostwick, Kerry A.


    In this paper, we question how we might disrupt positivist research paradigms that preclude students from engaging and experiencing ownership in the research process. We question what we, as professors, could do to facilitate the transition from traditional research reporting to a disposition of inquiry that allows for ambiguity and discovery in…

  12. Conscience in Childhood: Old Questions, New Answers (United States)

    Aksan, Nazan; Kochanska, Grazyna


    Although conscience has been the focus of reflection for centuries, fundamental questions regarding its organization have not been fully answered. To address those questions, the authors applied structural equation modeling techniques to longitudinal data comprising multiple behavioral measures of children's conscience, obtained in parallel…

  13. Fixed-Response Questions with a Difference. (United States)

    Johnstone, Alex H.; Ambusaidi, Abdullah


    Offers three types of fixed-response questions that are designed to overcome drawbacks appearing in the conventional forms of fixed-response questions such as not allowing the examiner to investigate reasoning, background, or prevent guessing. (Contains 14 references.) (Author/YDS)

  14. Pitfalls, prevention, and treatment of hyperuricemia during tumor lysis syndrome in the era of rasburicase (recombinant urate oxidase

    Directory of Open Access Journals (Sweden)

    Andrea Pession


    Full Text Available Andrea Pession, Fraia Melchionda, Claudia CastelliniOncologia Ematologia Pediatrica “Lalla Seràgnoli”, Clinica Pediatrica, Università degli Studi di Bologna, Bologna, ItalyAbstract: Along with hydration and urinary alkalinization, allopurinol has been the standard agent for the management of hyperuricemia in patients with a high tumor burden at risk of tumor lysis syndrome; however, this agent often fails to prevent and treat this complication effectively. Rasburicase (recombinant urate oxidase has been shown to be effective in reducing uric acid and preventing uric acid accumulation in patients with hematologic malignancies with hyperuricemia or at high risk of developing it. Rasburicase acts at the end of the purine catabolic pathway and, unlike allopurinol, does not induce accumulation of xanthine or hypoxanthine. Its rapid onset of action and the ability to lower pre-existing elevated uric acid levels are the advantages of rasburicase over allopurinol. Rasburicase represents an effective alternative to allopurinol to promptly reduce uric acid levels, improve patient’s electrolyte status, and reverse renal insufficiency. The drug, initially studied in pediatric patients with acute lymphoblastic leukemia and aggressive non-Hodgkin lymphoma, seems to show comparable benefit in adults with similar lymphoid malignancies or at high risk of tumor lysis syndrome. Current and future trials will evaluate alternative doses and different schedules of rasburicase to maintain its efficacy while reducing its cost. The review provides a comprehensive and detailed review of pathogenesis, laboratory, and clinical presentation of TLS together with clinical studies already performed both in pediatric and adult patients.Keywords: tumor lysis syndrome, urate oxidase, rasburicase, allopurinol, uric acid

  15. Analysis of students’ generated questions in laboratory learning environments

    Directory of Open Access Journals (Sweden)

    Juan Antonio Llorens-Molina


    Full Text Available In order to attain a reliable laboratory work assessment, we argue taking the Learning Environment as a core concept and a research paradigm that considers the factors affecting the laboratory as a particularly complex educational context. With regard to Laboratory Learning Environments (LLEs, a well known approach is the SLEI (Science Laboratory Environment Inventory. The aim of this research is to design and apply an alternative and qualitative assessment tool to characterize Laboratory Learning Environments in an introductory course of organic chemistry. An alternative and qualitative assessment tool would be useful for providing feed-back for experimental learning improvement; serving as a complementary triangulation tool in educational research on LLEs; and generating meaningful categories in order to design quantitative research instruments. Toward this end, spontaneous questions by students have been chosen as a reliable source of information. To process these questions, a methodology based on the Grounded Theory has been developed to provide a framework for characterizing LLEs. This methodology has been applied in two case studies. The conclusions lead us to argue for using more holistic assessment tools in both everyday practice and research. Likewise, a greater attention should be paid to metacognition to achieve suitable self-perception concerning students’ previous knowledge and manipulative skills.

  16. True or false: is that the question?

    Directory of Open Access Journals (Sweden)

    Abaúnza Chagín, María Claudia


    Full Text Available Objectives: Standard tests designed for medical students frequently include objective True/ False (T/F questions, in which there are only two possible answers that can be successfully reached at random. The purpose of this work was to demonstrate that asking for support of the answers diminishes the number of randomly correct ones, so improving the ability of questions to verify learnings. Methods: 8.188 T/F questions whose answers had to be supported; they were applied in 28 written tests of Pathology, at the undergraduate medical program (La Sabana University, Chía, Colombia. Answers were classified as correct or incorrect, and their support, as correct, partially correct and incorrect. In order to analyze the data, the kappa agreement coefficient between T/F questions and T/F questions with support was calculated, as well as per term and per test. Results: Out of 8.188 T/F questions, 6.112 (74.6% were correctly answered as true or false, 3.655 (44.6% had a correct support, and 2.336 (28.5% were not correctly supported, exhibiting a global agreement index of 0.378 and low agreement indexes per term. Conclusion: T/F questions could be accompanied by support of the answers in order to reduce the probability of randomly correct ones, to enhance the development of superior cognitive processes, and communicative essential competences.

  17. Socrates was not a pimp: changing the paradigm of questioning in medical education. (United States)

    Kost, Amanda; Chen, Frederick M


    The slang term "pimping" is widely recognized by learners and educators in the clinical learning environment as the act of more senior members of the medical team publicly asking questions of more junior members. Although questioning as a pedagogical practice has many benefits, pimping, as described in the literature, evokes negative emotions in learners and leads to an environment that is not conducive to adult learning. Medical educators may employ pimping as a pedagogic technique because of beliefs that it is a Socratic teaching method. Although problems with pimping have previously been identified, no alternative techniques for questioning in the clinical environment were suggested. The authors posit that using the term "pimping" to describe questioning in medical education is harmful and unprofessional, and they propose clearly defining pimping as "questioning with the intent to shame or humiliate the learner to maintain the power hierarchy in medical education." Explicitly separating pimping from the larger practice of questioning allows the authors to make three recommendations for improving questioning practices. First, educators should examine the purpose of each question they pose to learners. Second, they should apply historic and modern interpretations of Socratic teaching methods that promote critical thinking skills. Finally, they should consider adult learning theories to make concrete changes to their questioning practices. These changes can result in questioning that is more learner centered, aids in the acquisition of knowledge and skills, performs helpful formative and summative assessments of the learner, and improves community in the clinical learning environment.

  18. Partial characterization of polyphenol oxidase activity in raspberry fruits. (United States)

    González, E M; de Ancos, B; Cano, M P


    A partial characterization of polyphenol oxidase (PPO) activity in raspberry fruits is described. Two early cultivars harvested in May/June (Heritage and Autumm Bliss) and two late cultivars harvested in October-November (Ceva and Rubi) were analyzed for PPO activity. Stable and highly active PPO extracts were obtained using insoluble poly(vinylpyrrolidone) (PVP) and Triton X-100 in sodium phosphate, pH 7.0 buffer. Polyacrylamide gel electrophoresis of raspberry extracts under nondenaturing conditions resolved in one band (R(f)()(1) = 0.25). Raspberry PPO activity has pH optima of 8.0 and 5.5, both with catechol (0.1 M). Maximum activity was with D-catechin (catecholase activity), followed by p-coumaric acid (cresolase activity). Heritage raspberry also showed PPO activity toward 4-methylcatechol. Ceva and Autumm Bliss raspberries showed the higher PPO activity using catechol as substrate.

  19. [ROS and NADPH oxidase: key regulators of tumor vascularisation]. (United States)

    Garrido-Urbani, Sarah; Jaquet, Vincent; Imhof, Beat A


    Oxidative stress is the result of an imbalance between the production of reactive oxygen species (ROS) and antioxidant mechanisms. It is characterized by damage of all cellular components, DNA, proteins, lipids. ROS are nevertheless important for the physiology of an organism, as they are involved in the innate immune defense and several intracellular signaling pathways. They play an important role in tumorigenesis by promoting tumor vasculature, which is essential to their growth and metastatic processes. There are many sources of ROS in the cells, but the NOX enzymes (NADPH oxidase-dependent) are now recognized to have a major role in the oxidative stress process. Indeed, they are present in many tissues where their only function is to produce ROS. This article discusses the NOX in endothelial cells and their role in the tumor angiogenesis.

  20. Electrochemistry of xanthine oxidase and its interaction with nitric oxide. (United States)

    Zhou, Hui; Xu, Yi; Chen, Ting; Suzuki, Iwao; Li, Genxi


    With the help of nanocrystalline TiO2, the direct electrochemistry of xanthine oxidase (XOD) was achieved and two pairs of redox waves were observed. The interaction between XOD and nitric oxide (NO) was also investigated. The experimental results reveal that NO can be reduced at a XOD-nano TiO2 film modified electrode. When the NO concentration was low, the reduced product, HNO, would inactivate the protein. However, when the NO concentration was high, HNO would continue to react with NO to form N2O2- and N3O3-, which would not inhibit XOD, and thus the amount of active protein did not decrease any further.

  1. Melatonin activates the peroxidase-oxidase reaction and promotes oscillations. (United States)

    Olsen, L F; Lunding, A; Lauritsen, F R; Allegra, M


    We have studied the peroxidase-oxidase reaction with NADH and O2 as substrates and melatonin as a cofactor in a semibatch reactor. We show for the first time that melatonin is an activator of the reaction catalyzed by enzymes from both plant and animal sources. Furthermore, melatonin promotes oscillatory dynamics in the pH range from 5 to 6. The frequency of the oscillations depends on the pH such that an increase in pH was accompanied by a decrease in frequency. Conversely, an increase in the flow rate of NADH or an increase in the average concentration of NADH resulted in an increase in oscillation frequency. Complex dynamics were not observed with melatonin as a cofactor. These results are discussed in relation to observations of oscillatory dynamics and the function of melatonin and peroxidase in activated neutrophils.

  2. Process requirements of galactose oxidase catalyzed oxidation of alcohols

    DEFF Research Database (Denmark)

    Pedersen, Asbjørn Toftgaard; R. Birmingham, William; Rehn, Gustav


    biocatalyst for the oxidation of primary and secondary alcohols to their corresponding aldehydes and ketones, respectively. However, GOase requires a number of additives to sustain its catalytic function, such as the enzyme catalase for degradation of the byproduct hydrogen peroxide as well as single......-electron oxidants to reactivate the enzyme upon loss of the amino acid radical in its active site. In this work, the addition of catalase, single-electron oxidants, and copper ions was investigated systematically in order to find the minimum concentrations required to obtain a fully active GOase. Furthermore......, it was found that the concentration and type of buffer is essential for the activity of GOase, which was significantly more active in sodium phosphate buffer than in other buffers investigated. Enzyme stability and oxygen requirements are of crucial importance for the implementation of oxidase based processes...

  3. Polyphenol Oxidases in Crops: Biochemical, Physiological and Genetic Aspects

    Directory of Open Access Journals (Sweden)

    Francesca Taranto


    Full Text Available Enzymatic browning is a colour reaction occurring in plants, including cereals, fruit and horticultural crops, due to oxidation during postharvest processing and storage. This has a negative impact on the colour, flavour, nutritional properties and shelf life of food products. Browning is usually caused by polyphenol oxidases (PPOs, following cell damage caused by senescence, wounding and the attack of pests and pathogens. Several studies indicated that PPOs play a role in plant immunity, and emerging evidence suggested that PPOs might also be involved in other physiological processes. Genomic investigations ultimately led to the isolation of PPO homologs in several crops, which will be possibly characterized at the functional level in the near future. Here, focusing on the botanic families of Poaceae and Solanaceae, we provide an overview on available scientific literature on PPOs, resulting in useful information on biochemical, physiological and genetic aspects.

  4. Potential role of NADPH oxidase in pathogenesis of pancreatitis

    Institute of Scientific and Technical Information of China (English)

    Wei-Li; Cao; Xiao-Hui; Xiang; Kai; Chen; Wei; Xu; Shi-Hai; Xia


    Studies have demonstrated that reactive oxygen species(ROS) are closely related to inflammatory disorders. Nicotinamide adenine dinucleotide phosphate oxidase(NOX), originally found in phagocytes, is the main source of ROS in nonphagocytic cells. Besides directly producing the detrimental highly reactive ROS to act on biomolecules(lipids, proteins, and nucleic acids), NOX can also activate multiple signal transduction pathways, which regulate cell growth, proliferation, differentiation and apoptosis by producing ROS. Recently, research on pancreatic NOX is no longer limited to inflammatory cells, but extends to the aspect of pancreatic acinar cells and pancreatic stellate cells, which are considered to be potentially associated with pancreatitis. In this review, we summarize the literature on NOX protein structure, activation, function and its role in the pathogenesis of pancreatitis.

  5. Xanthine oxidase inhibitory activity of Hungarian wild-growing mushrooms. (United States)

    Ványolós, Attila; Orbán-Gyapai, Orsolya; Hohmann, Judit


    Mushrooms represent a remarkable and yet largely unexplored source of new, biologically active natural products. In this work, we report on the xanthine oxidase (XO) inhibitory activity of 47 wild-growing mushrooms native to Hungary. Aqueous and organic (n-hexane, chloroform, and 50% methanol) extracts of selected mushrooms from different families were screened for their XO inhibitory activities. Among the 188 extracts investigated, the chloroform and 50% methanol fractions proved to be the most effective. Some species exhibited high inhibitory activity, e.g., Hypholoma fasciculare (IC50  =67.76 ± 11.05 µg/mL), Suillus grevillei (IC50  =13.28 ± 1.58 µg/mL), and Tricholoma populinum (IC50  =85.08 ± 15.02 µg/mL); others demonstrated moderate or weak activity. Additional studies are warranted to characterize the compounds responsible for the XO inhibitory activity of mushroom extracts.

  6. Lysyl Oxidase, a Targetable Secreted Molecule Involved in Cancer Metastasis

    DEFF Research Database (Denmark)

    Cox, Thomas R; Gartland, Alison; Erler, Janine T


    Secondary metastatic cancer remains the single biggest cause of mortality and morbidity across most solid tumors. In breast cancer, 100% of deaths are attributed to metastasis. At present, there are no "cures" for secondary metastatic cancer of any form and there is an urgent unmet clinical need...... to improve the tools available in our arsenal against this disease, both in terms of treatment, but also prevention. Recently, we showed that hypoxic induction of the extracellular matrix modifying enzyme lysyl oxidase (LOX) correlates with metastatic dissemination to the bone in estrogen receptor negative...... breast cancer and is essential for the formation of premetastatic osteolytic lesions. We showed that in models of breast cancer metastasis, targeting LOX, or its downstream effects, significantly inhibited premetastatic niche formation and the resulting metastatic burden, offering preclinical validation...

  7. Molecular basis of infantile reversible cytochrome c oxidase deficiency myopathy. (United States)

    Horvath, Rita; Kemp, John P; Tuppen, Helen A L; Hudson, Gavin; Oldfors, Anders; Marie, Suely K N; Moslemi, Ali-Reza; Servidei, Serenella; Holme, Elisabeth; Shanske, Sara; Kollberg, Gittan; Jayakar, Parul; Pyle, Angela; Marks, Harold M; Holinski-Feder, Elke; Scavina, Mena; Walter, Maggie C; Coku, Jorida; Günther-Scholz, Andrea; Smith, Paul M; McFarland, Robert; Chrzanowska-Lightowlers, Zofia M A; Lightowlers, Robert N; Hirano, Michio; Lochmüller, Hanns; Taylor, Robert W; Chinnery, Patrick F; Tulinius, Mar; DiMauro, Salvatore


    Childhood-onset mitochondrial encephalomyopathies are usually severe, relentlessly progressive conditions that have a fatal outcome. However, a puzzling infantile disorder, long known as 'benign cytochrome c oxidase deficiency myopathy' is an exception because it shows spontaneous recovery if infants survive the first months of life. Current investigations cannot distinguish those with a good prognosis from those with terminal disease, making it very difficult to decide when to continue intensive supportive care. Here we define the principal molecular basis of the disorder by identifying a maternally inherited, homoplasmic m.14674T>C mt-tRNA(Glu) mutation in 17 patients from 12 families. Our results provide functional evidence for the pathogenicity of the mutation and show that tissue-specific mechanisms downstream of tRNA(Glu) may explain the spontaneous recovery. This study provides the rationale for a simple genetic test to identify infants with mitochondrial myopathy and good prognosis.

  8. Electron transfer rates and equilibrium within cytochrome c oxidase

    DEFF Research Database (Denmark)

    Farver, O; Einarsdóttir, O; Pecht, I


    identical within experimental error and independent of the enzyme concentration. This demonstrates that a fast intramolecular electron equilibration is taking place between CuA and heme a. The rate constants for CuA --> heme a ET and the reverse (heme a --> CuA) process were found to be 13 000 s-1 and 3700......Intramolecular electron transfer (ET) between the CuA center and heme a in bovine cytochrome c oxidase was investigated by pulse radiolysis. CuA, the initial electron acceptor, was reduced by 1-methyl nicotinamide radicals in a diffusion-controlled reaction, as monitored by absorption changes...... at 830 nm. After the initial reduction phase, the 830 nm absorption was partially restored, corresponding to reoxidation of the CuA center. Concomitantly, the absorption at 445 nm and 605 nm increased, indicating reduction of heme a. The rate constants for heme a reduction and CuA reoxidation were...

  9. Traumatic Brain Injury and NADPH Oxidase: A Deep Relationship

    Directory of Open Access Journals (Sweden)

    Cristina Angeloni


    Full Text Available Traumatic brain injury (TBI represents one of the major causes of mortality and disability in the world. TBI is characterized by primary damage resulting from the mechanical forces applied to the head as a direct result of the trauma and by the subsequent secondary injury due to a complex cascade of biochemical events that eventually lead to neuronal cell death. Oxidative stress plays a pivotal role in the genesis of the delayed harmful effects contributing to permanent damage. NADPH oxidases (Nox, ubiquitary membrane multisubunit enzymes whose unique function is the production of reactive oxygen species (ROS, have been shown to be a major source of ROS in the brain and to be involved in several neurological diseases. Emerging evidence demonstrates that Nox is upregulated after TBI, suggesting Nox critical role in the onset and development of this pathology. In this review, we summarize the current evidence about the role of Nox enzymes in the pathophysiology of TBI.


    Institute of Scientific and Technical Information of China (English)

    WANG Lingzhi; YUAN Hong; FANG Shibi; JIANG Yingyan


    The properties of immobilized glucose oxidase (GOD) by the complexes of diethylaminoethyl cellu -lose(DEAEC) with different polymers, such as polymethylacrylic acid (PMAA), polyacrylic acid (PAA), polystyrene sulfonic acid (PSSA), polyvinylalcohol (PVA), polyethylene oxide (PEO)and styrene-maleic acid copolymer (PSMA) were investigated. The activity of immobilized GOD was obviously influenced by the component of the DEAEC complexes. The relative activity of the immobilized GOD reached to maximum and over 90% of the native GOD. when the DEAEC-PMAA DEAEC-PAA complexes were used as a carrier with the molar ratio of DEAEC and polyacid of about one. Michaelis constants (Km) of the immobilized enzymes of DEAEC-GOD-PMAA and DEAEC-GOD-PAA were determined to be 1.25 and 1.00, respectively. Moreover, the immobilized GOD has a good storage stability and cyclic life.

  11. Docência no ensino superior: questões e alternativas = Teaching Higher Education: alternately questions

    Directory of Open Access Journals (Sweden)

    Fischer, Beatriz T. Daudt


    Full Text Available Há dois pressupostos aceitos comumente na academia, que se resumem nas seguintes proposições: a universidade é frequentada por adultos e adultos sabem o que querem, portanto, o professor no ensino universitário não precisa preocupar-se com pedagogias; e, para dar aula, o que o professor precisa é simplesmente dominar o conteúdo e ter uma boa comunicação. Diante destas duas afirmações, o artigo apresenta alguns contrapontos, alicerçados em estudos e pesquisas que aportam idéias contemporâneas acerca da dimensão epistemológica inerente aos processos de aprendizagem. São considerados também aspectos de base sociopolítica e cultural que interferem no contexto da sala de aula, bem como são desdobradas algumas alternativas para o cotidiano da sala de aula

  12. Molecular Dynamic Studies of the Complex Polyethylenimine and Glucose Oxidase

    Directory of Open Access Journals (Sweden)

    Beata Szefler


    Full Text Available Glucose oxidase (GOx is an enzyme produced by Aspergillus, Penicillium and other fungi species. It catalyzes the oxidation of β-d-glucose (by the molecular oxygen or other molecules, like quinones, in a higher oxidation state to form d-glucono-1,5-lactone, which hydrolyses spontaneously to produce gluconic acid. A coproduct of this enzymatic reaction is hydrogen peroxide (H2O2. GOx has found several commercial applications in chemical and pharmaceutical industries including novel biosensors that use the immobilized enzyme on different nanomaterials and/or polymers such as polyethylenimine (PEI. The problem of GOx immobilization on PEI is retaining the enzyme native activity despite its immobilization onto the polymer surface. Therefore, the molecular dynamic (MD study of the PEI ligand (C14N8_07_B22 and the GOx enzyme (3QVR was performed to examine the final complex PEI-GOx stabilization and the affinity of the PEI ligand to the docking sites of the GOx enzyme. The docking procedure showed two places/regions of major interaction of the protein with the polymer PEI: (LIG1 of −5.8 kcal/mol and (LIG2 of −4.5 kcal/mol located inside the enzyme and on its surface, respectively. The values of enthalpy for the PEI-enzyme complex, located inside of the protein (LIG1 and on its surface (LIG2 were computed. Docking also discovered domains of the GOx protein that exhibit no interactions with the ligand or have even repulsive characteristics. The structural data clearly indicate some differences in the ligand PEI behavior bound at the two places/regions of glucose oxidase.

  13. Putrescine biosensor based on putrescine oxidase from Kocuria rosea. (United States)

    Bóka, Beáta; Adányi, Nóra; Szamos, Jenő; Virág, Diána; Kiss, Attila


    The novel putrescine oxidase based amperometric biosensor selectively measures putrescine, which can be considered as an indicator of microbial spoilage. Putrescine oxidase (PUOX, EC was isolated from Kocuria rosea (Micrococcus rubens) by an improved and simplified purification process. Cells were grown on brain heart infusion medium supplemented with putrescine. Cell-free extract was prepared in Tris buffer (pH 8.0) by Bead-beater. A newly elaborated step based on three-phase partitioning (TPP) was applied in the purification protocol of PUOX. The purified enzyme was immobilized on the surface of a spectroscopic graphite electrode in redox hydrogel with horseradish peroxidase, Os mediator and poly(ethylene glycol) (400) diglycidyl ether (PEGDGE) as crosslinking agent. This modified working electrode was used in wall-jet type amperometric cell together with the Ag/AgCl (0.1M KCl) reference electrode and a platinum wire as auxiliary electrode in flow injection analysis system (FIA). Hydrogel composition, pH and potential dependence were studied. Optimal working conditions were 0.45 mLmin(-1) flow rate of phosphate buffer (66 mM, pH 8.0) and +50 mV polarizing potential vs. Ag/AgCl. The linear measuring range of the method was 0.01-0.25 mM putrescine, while the detection limit was 5 μM. Beer samples were investigated by the putrescine biosensor and the results were compared by those of HPLC reference method.

  14. Role of NADPH Oxidase-4 in Human Endothelial Progenitor Cells (United States)

    Hakami, Nora Y.; Ranjan, Amaresh K.; Hardikar, Anandwardhan A.; Dusting, Greg J.; Peshavariya, Hitesh M.


    Introduction: Endothelial progenitor cells (EPCs) display a unique ability to promote angiogenesis and restore endothelial function in injured blood vessels. NADPH oxidase 4 (NOX4)-derived hydrogen peroxide (H2O2) serves as a signaling molecule and promotes endothelial cell proliferation and migration as well as protecting against cell death. However, the role of NOX4 in EPC function is not completely understood. Methods: EPCs were isolated from human saphenous vein and mammary artery discarded during bypass surgery. NOX4 gene and protein expression in EPCs were measured by real time-PCR and Western blot analysis respectively. NOX4 gene expression was inhibited using an adenoviral vector expressing human NOX4 shRNA (Ad-NOX4i). H2O2 production was measured by Amplex red assay. EPC migration was evaluated using a transwell migration assay. EPC proliferation and viability were measured using trypan blue counts. Results: Inhibition of NOX4 using Ad-NOX4i reduced Nox4 gene and protein expression as well as H2O2 formation in EPCs. Inhibition of NOX4-derived H2O2 decreased both proliferation and migration of EPCs. Interestingly, pro-inflammatory cytokine tumor necrosis factor alpha (TNFα) decreased NOX4 expression and reduced survival of EPCs. However, the survival of EPCs was further diminished by TNF-α in NOX4-knockdown cells, suggesting that NOX4 has a protective role in EPCs. Conclusion: These findings suggest that NOX4-type NADPH oxidase is important for proliferation and migration functions of EPCs and protects against pro-inflammatory cytokine induced EPC death. These properties of NOX4 may facilitate the efficient function of EPCs which is vital for successful neovascularization.

  15. Molecular Dynamic Studies of the Complex Polyethylenimine and Glucose Oxidase (United States)

    Szefler, Beata; Diudea, Mircea V.; Putz, Mihai V.; Grudzinski, Ireneusz P.


    Glucose oxidase (GOx) is an enzyme produced by Aspergillus, Penicillium and other fungi species. It catalyzes the oxidation of β-d-glucose (by the molecular oxygen or other molecules, like quinones, in a higher oxidation state) to form d-glucono-1,5-lactone, which hydrolyses spontaneously to produce gluconic acid. A coproduct of this enzymatic reaction is hydrogen peroxide (H2O2). GOx has found several commercial applications in chemical and pharmaceutical industries including novel biosensors that use the immobilized enzyme on different nanomaterials and/or polymers such as polyethylenimine (PEI). The problem of GOx immobilization on PEI is retaining the enzyme native activity despite its immobilization onto the polymer surface. Therefore, the molecular dynamic (MD) study of the PEI ligand (C14N8_07_B22) and the GOx enzyme (3QVR) was performed to examine the final complex PEI-GOx stabilization and the affinity of the PEI ligand to the docking sites of the GOx enzyme. The docking procedure showed two places/regions of major interaction of the protein with the polymer PEI: (LIG1) of −5.8 kcal/mol and (LIG2) of −4.5 kcal/mol located inside the enzyme and on its surface, respectively. The values of enthalpy for the PEI-enzyme complex, located inside of the protein (LIG1) and on its surface (LIG2) were computed. Docking also discovered domains of the GOx protein that exhibit no interactions with the ligand or have even repulsive characteristics. The structural data clearly indicate some differences in the ligand PEI behavior bound at the two places/regions of glucose oxidase. PMID:27801788

  16. RETRACTED: Protonation State-Dependent Communication in Cytochrome c Oxidase. (United States)

    Helabad, Mahdi Bagherpoor; Ghane, Tahereh; Reidelbach, Marco; Woelke, Anna Lena; Knapp, Ernst Walter; Imhof, Petra


    Proton transfer in cytochrome c oxidase from the cellular inside to the binuclear redox center (BNC) can occur through two distinct pathways, the D- and K-channels. For the protein to function as both redox enzyme and proton pump, proton transfer out of either of the channels toward the BNC or into the protein toward a proton loading site, and ultimately through the membrane, must be highly regulated. The O→E intermediate of cytochrome c oxidase is the first redox state in its catalytic cycle, where proton transfer through the K-channel, from K362 to Y288 at the BNC, is important. Molecular dynamics simulations of this intermediate with 16 different combinations of protonation states of key residues in the D- and K-channel show the mutual impact of the two proton-conducting channels to be protonation state-dependent. Strength as well as means of communication, correlations in positions, or connections along the hydrogen-bonded network, change with the protonation state of the K-channel residue K362. The conformational and hydrogen-bond dynamics of the D-channel residue N139 regulated by an interplay of protonation in the D-channel and K362. N139 thus assumes a gating function by which proton passage through the D-channel toward E286 is likely facilitated for states with protonated K362 and unprotonated E286, which would in principle allow proton transfer to the BNC, but no proton pumping until a proton has reached E286. Copyright © 2016 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  17. Molecular Dynamic Studies of the Complex Polyethylenimine and Glucose Oxidase. (United States)

    Szefler, Beata; Diudea, Mircea V; Putz, Mihai V; Grudzinski, Ireneusz P


    Glucose oxidase (GOx) is an enzyme produced by Aspergillus, Penicillium and other fungi species. It catalyzes the oxidation of β-d-glucose (by the molecular oxygen or other molecules, like quinones, in a higher oxidation state) to form d-glucono-1,5-lactone, which hydrolyses spontaneously to produce gluconic acid. A coproduct of this enzymatic reaction is hydrogen peroxide (H₂O₂). GOx has found several commercial applications in chemical and pharmaceutical industries including novel biosensors that use the immobilized enzyme on different nanomaterials and/or polymers such as polyethylenimine (PEI). The problem of GOx immobilization on PEI is retaining the enzyme native activity despite its immobilization onto the polymer surface. Therefore, the molecular dynamic (MD) study of the PEI ligand (C14N8_07_B22) and the GOx enzyme (3QVR) was performed to examine the final complex PEI-GOx stabilization and the affinity of the PEI ligand to the docking sites of the GOx enzyme. The docking procedure showed two places/regions of major interaction of the protein with the polymer PEI: (LIG1) of -5.8 kcal/mol and (LIG2) of -4.5 kcal/mol located inside the enzyme and on its surface, respectively. The values of enthalpy for the PEI-enzyme complex, located inside of the protein (LIG1) and on its surface (LIG2) were computed. Docking also discovered domains of the GOx protein that exhibit no interactions with the ligand or have even repulsive characteristics. The structural data clearly indicate some differences in the ligand PEI behavior bound at the two places/regions of glucose oxidase.


    Directory of Open Access Journals (Sweden)

    Gunawan Gunawan


    Full Text Available Question answering system is a system that allows user to state his or her information need in the form of natural language question, and return short text excerpts or even phrases as an answer. The availability of a wide and various information source and improvements in the techniques of natural language processing, information extraction (wrapper, and information retrieval give a big effect on the development of question answering system, from just answering questions in a specific domain by consulting to structured information source such as database, and like in this research, answering any questions based on information stored in an unstructured text collection. A general architecture of question answering system based on text consists of six processing stages, i.e. question analysis, document collection preprocessing, candidate document selection, candidate document analysis, answer extraction, and response generation. Application of question answering system like AnswerBus, Mulder, and Webclopedia that are developed with its own characteristics has similar processing steps as in the general architecture. Answers returned by a question answering system need to be evaluated for performance measure. This research completed with a simple question answering system application using english Bible in World English Bible (WEB version as the source of information to answer some questions. Because specific domain is selected: Bible, questions that can be posed by user could ask about information in the Bible itself only. Question is also limited to three types of answers that can be supported by the application: person (who, location (where, and date (when. Abstract in Bahasa Indonesia : Question answering system (QA system adalah sistem yang mengijinkan user menyatakan kebutuhan informasinya dalam bentuk natural language question (pertanyaan dalam bahasa alami, dan mengembalikan kutipan teks singkat atau bahkan frase sebagai jawaban. Ketersediaan

  19. Skeletal muscle NADPH oxidase is increased and triggers stretch-induced damage in the mdx mouse. (United States)

    Whitehead, Nicholas P; Yeung, Ella W; Froehner, Stanley C; Allen, David G


    Recent studies have shown that oxidative stress contributes to the pathogenesis of muscle damage in dystrophic (mdx) mice. In this study we have investigated the role of NADPH oxidase as a source of the oxidative stress in these mice. The NADPH oxidase subunits gp91(phox), p67(phox) and rac 1 were increased 2-3 fold in tibilais anterior muscles from mdx mice compared to wild type. Importantly, this increase occurred in 19 day old mice, before the onset of muscle necrosis and inflammation, suggesting that NADPH oxidase is an important source of oxidative stress in mdx muscle. In muscles from 9 week old mdx mice, gp91(phox) and p67(phox) were increased 3-4 fold and NADPH oxidase superoxide production was 2 times greater than wild type. In single fibers from mdx muscle NADPH oxidase subunits were all located on or near the sarcolemma, except for p67(phox),which was expressed in the cytosol. Pharmacological inhibition of NADPH oxidase significantly reduced the intracellular Ca(2+) rise following stretched contractions in mdx single fibers, and also attenuated the loss of muscle force. These results suggest that NADPH oxidase is a major source of reactive oxygen species in dystrophic muscle and its enhanced activity has a stimulatory effect on stretch-induced Ca(2+) entry, a key mechanism for muscle damage and functional impairment.

  20. Cytochrome c oxidase-intermediate fibres: importance in understanding the pathogenesis and treatment of mitochondrial myopathy. (United States)

    Murphy, Julie L; Ratnaike, Thiloka E; Shang, Ersong; Falkous, Gavin; Blakely, Emma L; Alston, Charlotte L; Taivassalo, Tanja; Haller, Ronald G; Taylor, Robert W; Turnbull, Doug M


    An important diagnostic muscle biopsy finding in patients with mitochondrial DNA disease is the presence of respiratory-chain deficient fibres. These fibres are detected as cytochrome c oxidase-deficient following a sequential cytochrome c oxidase-succinate dehydrogenase reaction, often in a mosaic pattern within a population of cytochrome c oxidase-normal fibres. Detailed analysis of muscle biopsies from patients with various mitochondrial DNA defects shows that a spectrum of deficiency exists, as there are a large number of fibres which do not correspond to being either completely cytochrome c oxidase-normal (brown staining) or cytochrome c oxidase-deficient (blue staining). We have used a combination of histochemical and immunocytochemical techniques to show that a population of cytochrome c oxidase-intermediate reacting fibres are a gradation between normal and deficient fibres. We show that cytochrome c oxidase-intermediate fibres also have different genetic characteristics in terms of amount of mutated and wild-type mtDNA, and as such, may represent an important transition between respiratory normal and deficient fibres. Assessing changes in intermediate fibres will be crucial to evaluating the responses to treatment and in particular to exercise training regimes in patients with mitochondrial DNA disease.

  1. Frequently Asked Questions on Potassium Iodide (KI) (United States)

    ... Bioterrorism and Drug Preparedness Frequently Asked Questions on Potassium Iodide (KI) Share Tweet Linkedin Pin it More sharing ... Drug Administration (FDA) issued a final Guidance on Potassium Iodide as a Thyroid Blocking Agent in Radiation Emergencies) ( ...

  2. 101 questions on corals: Towards awareness

    Digital Repository Service at National Institute of Oceanography (India)

    Wafar, M.V.M.; Wafar, S.

    to nature and man-made changes, in every stakeholder, from local population through tourists to managers. This book serves such a purpose by answering most of the questions we ask everyday about corals and reefs....

  3. Questions and Answers: Apple Juice and Arsenic (United States)

    ... Home Food Resources for You Consumers Questions & Answers: Apple Juice and Arsenic Share Tweet Linkedin Pin it ... MMA), may also be a health concern. Are apple and other fruit juices safe to drink? The ...

  4. Frequently Asked Questions about Digital Mammography (United States)

    ... Consumer Information (MQSA) Frequently Asked Questions About Digital Mammography Share Tweet Linkedin Pin it More sharing options ... mammography exams, such as DBT? What is digital mammography? Full field digital mammography (FFDM, also known simply ...

  5. Frequently Asked Questions about Genetic Testing (United States)

    ... Care Specific Genetic Disorders Frequently Asked Questions About Genetic Testing What is genetic testing? What can I learn ... find more information about genetic testing? What is genetic testing? Genetic testing uses laboratory methods to look at ...

  6. Frequently Asked Questions about Genetic Counseling (United States)

    ... on this page Frequently Asked Questions About Genetic Counseling What are genetic professionals and what do they ... genetics nurses. Top of page What is genetic counseling and evaluation? Genetic professionals work as members of ...

  7. FAQ: General Questions about West Nile Virus (United States)

    ... Public Service Videos General Questions About West Nile Virus Recommend on Facebook Tweet Share Compartir On this ... West Nile virus cases? What is West Nile virus? West Nile virus is an arthropod-borne virus ( ...

  8. The Case for Alternative Social Media

    Directory of Open Access Journals (Sweden)

    Robert W. Gehl


    Full Text Available What are “alternative social media”? How can we distinguish alternative social media from mainstream social media? Why are social media alternatives important? How do they work? Why do people make them? What do they tell us about contemporary corporate social media and its related phenomena: surveillance, privacy, power, self-expression, and sociality? This essay answers these questions by theorizing alternative social media. The empirical data for this alternative social media theory are drawn from previous work on alternative sites such as Diaspora,, Twister, GNU social, and the Dark Web Social Network. These cases are used to build a generalized conceptual framework. However, this article does not solely theorize from these examples, but rather seeks to contextualize and historicize alternative social media theory within larger bodies of work. In addition to generalization from examples, the theory is informed by two threads. The first thread is the work of alternative media scholars such as Nick Couldry, Chris Atton, and Clemencia Rodriguez, who have done the historical and theoretical work to define alternative media. The second thread is a synthesis of works exploring the technical side of contemporary media, coming from new fields such as software studies. The threads and empirical analyses of sites such as Diaspora, Quitter, and are combined into a theoretical matrix that can account for the processes and technical infrastructures that comprise social media alternatives and explain why they are distinct from sites such as Facebook, Twitter, and Google, as well as why they are important.

  9. Synthesis and Biological Evaluation of Novel Aryl-2H-pyrazole Derivatives as Potent Non-purine Xanthine Oxidase Inhibitors. (United States)

    Sun, Zhi-Gang; Zhou, Xiao-Jing; Zhu, Ming-Li; Ding, Wen-Ze; Li, Zhen; Zhu, Hai-Liang


    A series of aryl-2H-pyrazole derivatives were synthesized and evaluated for inhibitory activity against xanthine oxidase in vitro as potent xanthine oxidase inhibitors. Among them, 2 aryl-2H-pyrazole derivatives showed significant inhibitory activities against xanthine oxidase. Compound 19 emerged as the most potent xanthine oxidase inhibitor (IC50=9.8 µM) in comparison with allopurinol (IC50=9.5 µM). The docking study revealed that compound 19 might have strong interactions with the active site of xanthine oxidase. This compound is thus a new candidate for further development for the treatment of gout.

  10. Alternative Solar Indices

    Energy Technology Data Exchange (ETDEWEB)

    Lantz, L.J.


    Possible alternative Solar Indices which could either be a perturbation from the currently defined Solar Index or possible indices based on current technologies for other media markets are discussed. An overview is given of the current project, including the logic that was utilized in defining its current structure and then alternative indices and definitions are presented and finally, recommendations are made for adopting alternative indices.

  11. On alternating quantum walks (United States)

    Rousseva, Jenia; Kovchegov, Yevgeniy


    We study an inhomogeneous quantum walk on a line that evolves according to alternating coins, each a rotation matrix. For the quantum walk with the coin alternating between clockwise and counterclockwise rotations by the same angle, we derive a closed form solution for the propagation of probabilities, and provide its asymptotic approximation via the method of stationary phase. Finally, we observe that for a x03c0;/4 angle, this alternating rotation walk will replicate the renown Hadamard walk.

  12. On an Alternative Cosmology

    CERN Document Server

    Vankov, A


    The suggested alternative cosmology is based on the idea of barion symmetric universe, in which our home universe is a representative of multitude of typical matter and antimatter universes. This alternative concept gives a physically reasonable explanation of all major problems of the Standard Cosmological Model. Classification Code MSC: Cosmology 524.8 Key words: standard cosmological model, alternative cosmology, barionic symmetry, typical universe, quasars, cosmic rays.


    Institute of Scientific and Technical Information of China (English)


    Tag questions are a distinctive feature in spokenEnglish,but seldom used by the non-native speakers.Studies show that tags are employed by native speakersmainly to express their interactive and interpersonalmeaning.In this paper,the writer explores the natureof tag questions,mainly their interactive and inter-personal functions in daily coversation.Suggestionsare made with regard to the teaching of tags in theclassroom.

  14. Open-ended visual question answering


    Masuda Mora, Issey


    Wearable cameras generate a large amount of photos which are, in many cases, useless or redundant. On the other hand, these devices are provide an excellent opportunity to create automatic questions and answers for reminiscence therapy. This is a follow up of the BSc thesis developed by Ricard Mestre during Fall 2014, and MSc thesis developed by Aniol Lidon. This thesis studies methods to solve Visual Question-Answering (VQA) tasks with a Deep Learning framework. As a preliminary step, we ...

  15. Statin intolerance: more questions than answers. (United States)

    Guyton, John R; Campbell, Kristen B; Lakey, Wanda C


    The dramatic effectiveness of statins in improving the course of atherosclerotic cardiovascular disease tends to overshadow questions of statin intolerance. Thus after more than 25 years of clinical statin use, intolerance remains a poorly understood, frustrating issue for patients and providers. It has been extraordinarily difficult to define statin intolerance and its implications for clinical practice. Here, we briefly summarize current knowledge and raise questions that need to be addressed.



    Nelu DORLE


    Making a good sales approach depends largely on the strategy established by a salesperson, depending on the type of customer, the circumstances, and his/her psycho-linguistic availability. The sales strategies based on the science of reasoning, on the oratory and persuasive ability include skills related to communication, on which one of the most important is the development and asking of questions. The science related to the salesperson’s ability to handles questions in a sales interview giv...

  17. Four Principles for Selecting HCI Research Questions

    DEFF Research Database (Denmark)

    Clemmensen, Torkil

    In this position paper, I present and explain the position that what we should study in HCI depends on the objective of the research and its political, social, cultural, technological, and historical context. I outline four principles for selecting research questions and give a personal account...... of how I have selected research questions using these four principles. The aim with the paper is to generate discussion and advance the understanding of what to study in HCI....

  18. Alternative medicine studies

    National Research Council Canada - National Science Library


    A peer-reviewed, open-access journal about alternative medicine systems including acupuncture, anthroposophic medicine, ayurveda, chiropractic, herbalism and natural products, homeopathy, naturopathy...

  19. Alternative Auditing Approaches

    Energy Technology Data Exchange (ETDEWEB)

    Kandt, Alicen J [National Renewable Energy Laboratory (NREL), Golden, CO (United States)


    This presentation for the 2017 Energy Exchange in Tampa, Florida, offers information about advanced auditing technologies and techniques including alternative auditing approaches and considerations and caveats.

  20. Brandmodstandsbidrag for alternative isoleringsmaterialer

    DEFF Research Database (Denmark)

    Hansen, Ernst Jan de Place


    Resume af rapport om alternative isoleringsmaterialers brandmodstandsbidrag, udarbejdet af Dansk Brandteknisk Institut under Energistyrelsens udviklingsprogram "Miljø- og arbejdsmiljøvenlig isolering"...